Susan Campbell

Chronic kidney disease (CKD) is a significant public health problem, and recent genetic studies have identified common CKD susceptibility variants. The CKDGen consortium performed a meta-analysis of genome-wide association data in 67,093 individuals of European ancestry from 20 predominantly population-based studies in order to identify new susceptibility loci for reduced renal function as estimated by serum creatinine (eGFRcrea), serum cystatin c (eGFRcys) and CKD (eGFRcrea < 60 ml/min/1.73 m(2); n = 5,807 individuals with CKD (cases)). Follow-up of the 23 new genome-wide-significant loci (P < 5 x 10(-8)) in 22,982 replication samples identified 13 new loci affecting renal function and CKD (in or near LASS2, GCKR, ALMS1, TFDP2, DAB2, SLC34A1, VEGFA, PRKAG2, PIP5K1B, ATXN2, DACH1, UBE2Q2 and SLC7A9) and 7 loci suspected to affect creatinine production and secretion (CPS1, SLC22A2, TMEM60, WDR37, SLC6A13, WDR72 and BCAS3). These results further our understanding of the biologic mechanisms of kidney function by identifying loci that potentially influence nephrogenesis, podocyte function, angiogenesis, solute transport and metabolic functions of the kidney.

Anna Köttgen and colleagues report genome-wide association studies for serum urate in over 140,000 individuals from the Global Urate Genetics Consortium (GUGC). They identify 18 loci newly associated with serum urate concentrations and confirm 10 known loci, characterize their associations with gout and include a network analysis suggesting a role for inhibins-activins pathways in regulating urate homeostasis. Elevated serum urate concentrations can cause gout, a prevalent and painful inflammatory arthritis. By combining data from >140,000 individuals of European ancestry within the Global Urate Genetics Consortium (GUGC), we identified and replicated 28 genome-wide significant loci in association with serum urate concentrations (18 new regions in or near TRIM46, INHBB, SFMBT1, TMEM171, VEGFA, BAZ1B, PRKAG2, STC1, HNF4G, A1CF, ATXN2, UBE2Q2, IGF1R, NFAT5, MAF, HLF, ACVR1B-ACVRL1 and B3GNT4). Associations for many of the loci were of similar magnitude in individuals of non-European ancestry. We further characterized these loci for associations with gout, transcript expression and the fractional excretion of urate. Network analyses implicate the inhibins-activins signaling pathways and glucose metabolism in systemic urate control. New candidate genes for serum urate concentration highlight the importance of metabolic control of urate production and excretion, which may have implications for the treatment and prevention of gout.

Jonathan Marchini and colleagues with the Ox-GSK consortium report a meta-analysis for smoking phenotypes from 20 studies including 41,150 individuals, confirming an association at the CHRNA5–CHRNA3 locus on 15q25 to smoking quantity. They use imputation based on 1,000 Genomes Project Pilot 1 data to refine the association at this locus. Smoking is a leading global cause of disease and mortality1. We established the Oxford-GlaxoSmithKline study (Ox-GSK) to perform a genome-wide meta-analysis of SNP association with smoking-related behavioral traits. Our final data set included 41,150 individuals drawn from 20 disease, population and control cohorts. Our analysis confirmed an effect on smoking quantity at a locus on 15q25 (P = 9.45 × 10−19) that includes CHRNA5, CHRNA3 and CHRNB4, three genes encoding neuronal nicotinic acetylcholine receptor subunits. We used data from the 1000 Genomes project to investigate the region using imputation, which allowed for analysis of virtually all common SNPs in the region and offered a fivefold increase in marker density over HapMap2 (ref. 2) as an imputation reference panel. Our fine-mapping approach identified a SNP showing the highest significance, rs55853698, located within the promoter region of CHRNA5. Conditional analysis also identified a secondary locus (rs6495308) in CHRNA3.

The ACMEplus project aims to devise a standardised system for measuring case-mix and outcome in older patients admitted to hospitals in different parts of Europe for primarily 'medical' (i.e. not surgical or psychiatric) reasons. As a first step in this project, a systematic review was carried out to identify factors which had a significant influence on outcome in such patients.The systematic search used Medline 1966-2000, Cinahl 1982-2000, Web of Science 1981-2000, reference lists of relevant papers and a hand search of Age and Ageing 1974-2000. A six-category grading system was devised to classify the 313 identified papers with regard to their relevance to the ACMEplus project, study design and power. The analysis of the 14 'category 1' papers is presented.The main areas of assessment of case-mix were function, cognition, depression, illness severity, nutrition, social elements, aspects of diagnosis and demographic details. Statistically significant predictors, for the four outcome measures, listed below were: For length of stay: functional status score, illness severity, cognitive score, poor nutrition, comorbidity score, diagnosis or presenting illness, polypharmacy, age and gender. For mortality: functional status score, illness severity, cognitive score, comorbidity score, diagnosis or presenting illness, polypharmacy, age and gender. For discharge destination: functional status score, cognitive score, diagnosis or presenting illness and age. For readmission rate: functional status score, illness severity, co-morbidity, polypharmacy, diagnosis or presenting illness and age.Factors affecting outcome in older medical patients are complex. When looking at outcomes of hospital admission in older people it is important not just to look at routinely available statistics such as age, gender and diagnosis but also to take into account multifaceted aspects such as functional status and cognitive function.

Over half of all proteins are glycosylated, and alterations in glycosylation have been observed in numerous physiological and pathological processes. Attached glycans significantly affect protein function; but, contrary to polypeptides, they are not directly encoded by genes, and the complex processes that regulate their assembly are poorly understood. A novel approach combining genome-wide association and high-throughput glycomics analysis of 2,705 individuals in three population cohorts showed that common variants in the Hepatocyte Nuclear Factor 1α (HNF1α) and fucosyltransferase genes FUT6 and FUT8 influence N-glycan levels in human plasma. We show that HNF1α and its downstream target HNF4α regulate the expression of key fucosyltransferase and fucose biosynthesis genes. Moreover, we show that HNF1α is both necessary and sufficient to drive the expression of these genes in hepatic cells. These results reveal a new role for HNF1α as a master transcriptional regulator of multiple stages in the fucosylation process. This mechanism has implications for the regulation of immunity, embryonic development, and protein folding, as well as for our understanding of the molecular mechanisms underlying cancer, coronary heart disease, and metabolic and inflammatory disorders.

Urinary incontinence is common after radical prostatectomy and can also occur in some circumstances after transurethral resection of the prostate (TURP). Conservative management includes pelvic floor muscle training with or without biofeedback, electrical stimulation, extra-corporeal magnetic innervation (ExMI), compression devices (penile clamps), lifestyle changes, or a combination of methods.To determine the effectiveness of conservative management for urinary incontinence up to 12 months after transurethral, suprapubic, laparoscopic, radical retropubic or perineal prostatectomy, including any single conservative therapy or any combination of conservative therapies.We searched the Cochrane Incontinence Group Specialised Register (5 February 2014), CENTRAL (2014, Issue 1), EMBASE (January 2010 to Week 3 2014), CINAHL (January 1982 to 18 January 2014), ClinicalTrials.gov and World Health Organization (WHO) International Clinical Trials Registry Platform (ICTRP) (both searched 29 January 2014), and the reference lists of relevant articles.Randomised or quasi-randomised controlled trials evaluating conservative interventions for urinary continence in men after prostatectomy.Two or more review authors assessed the methodological quality of the trials and abstracted data. We tried to contact several authors of included studies to obtain extra information.Fifty trials met the inclusion criteria, 45 in men after radical prostatectomy, four trials after TURP and one trial after either operation. The trials included 4717 men of whom 2736 had an active conservative intervention. There was considerable variation in the interventions, populations and outcome measures. Data were not available for many of the pre-stated outcomes. Men's symptoms improved over time irrespective of management.There was no evidence from eight trials that pelvic floor muscle training with or without biofeedback was better than control for men who had urinary incontinence up to 12 months after radical prostatectomy; the quality of the evidence was judged to be moderate (for example 57% with urinary incontinence in the intervention group versus 62% in the control group, risk ratio (RR) for incontinence after 12 months 0.85, 95% confidence interval (CI) 0.60 to 1.22). One large multi-centre trial of one-to-one therapy showed no difference in any urinary or quality of life outcome measures and had narrow CIs. It seems unlikely that men benefit from one-to-one PFMT therapy after TURP. Individual small trials provided data to suggest that electrical stimulation, external magnetic innervation, or combinations of treatments might be beneficial but the evidence was limited. Amongst trials of conservative treatment for all men after radical prostatectomy, aimed at both treatment and prevention, there was moderate evidence of an overall benefit from pelvic floor muscle training versus control management in terms of reduction of urinary incontinence (for example 10% with urinary incontinence after one year in the intervention groups versus 32% in the control groups, RR for urinary incontinence 0.32, 95% CI 0.20 to 0.51). However, this finding was not supported by other data from pad tests. The findings should be treated with caution because the risk of bias assessment showed methodological limitations. Men in one trial were more satisfied with one type of external compression device, which had the lowest urine loss, compared to two others or no treatment. The effect of other conservative interventions such as lifestyle changes remained undetermined as no trials involving these interventions were identified.The value of the various approaches to conservative management of postprostatectomy incontinence after radical prostatectomy remains uncertain. The evidence is conflicting and therefore rigorous, adequately powered randomised controlled trials (RCTs) which abide by the principles and recommendations of the CONSORT statement are still needed to obtain a definitive answer. The trials should be robustly designed to answer specific well constructed research questions and include outcomes which are important from the patient's perspective in decision making and are also relevant to the healthcare professionals. Long-term incontinence may be managed by an external penile clamp, but there are safety problems.

Phospho- and sphingolipids are crucial cellular and intracellular compounds. These lipids are required for active transport, a number of enzymatic processes, membrane formation, and cell signalling. Disruption of their metabolism leads to several diseases, with diverse neurological, psychiatric, and metabolic consequences. A large number of phospholipid and sphingolipid species can be detected and measured in human plasma. We conducted a meta-analysis of five European family-based genome-wide association studies (N = 4034) on plasma levels of 24 sphingomyelins (SPM), 9 ceramides (CER), 57 phosphatidylcholines (PC), 20 lysophosphatidylcholines (LPC), 27 phosphatidylethanolamines (PE), and 16 PE-based plasmalogens (PLPE), as well as their proportions in each major class. This effort yielded 25 genome-wide significant loci for phospholipids (smallest P-value = 9.88×10−204) and 10 loci for sphingolipids (smallest P-value = 3.10×10−57). After a correction for multiple comparisons (P-value<2.2×10−9), we observed four novel loci significantly associated with phospholipids (PAQR9, AGPAT1, PKD2L1, PDXDC1) and two with sphingolipids (PLD2 and APOE) explaining up to 3.1% of the variance. Further analysis of the top findings with respect to within class molar proportions uncovered three additional loci for phospholipids (PNLIPRP2, PCDH20, and ABDH3) suggesting their involvement in either fatty acid elongation/saturation processes or fatty acid specific turnover mechanisms. Among those, 14 loci (KCNH7, AGPAT1, PNLIPRP2, SYT9, FADS1-2-3, DLG2, APOA1, ELOVL2, CDK17, LIPC, PDXDC1, PLD2, LASS4, and APOE) mapped into the glycerophospholipid and 12 loci (ILKAP, ITGA9, AGPAT1, FADS1-2-3, APOA1, PCDH20, LIPC, PDXDC1, SGPP1, APOE, LASS4, and PLD2) to the sphingolipid pathways. In large meta-analyses, associations between FADS1-2-3 and carotid intima media thickness, AGPAT1 and type 2 diabetes, and APOA1 and coronary artery disease were observed. In conclusion, our study identified nine novel phospho- and sphingolipid loci, substantially increasing our knowledge of the genetic basis for these traits.

Background— Hutchinson-Gilford progeria syndrome is an ultrarare segmental premature aging disease resulting in early death from heart attack or stroke. There is no approved treatment, but starting in 2007, several recent single-arm clinical trials administered inhibitors of protein farnesylation aimed at reducing toxicity of the disease-producing protein progerin. No study assessed whether treatments influence patient survival. The key elements necessary for this analysis are a robust natural history of survival and comparison with a sufficiently large patient population that has been treated for a sufficient time period with disease-targeting medications. Methods and Results— We generated Kaplan–Meier survival analyses for the largest untreated Hutchinson-Gilford progeria syndrome cohort to date. Mean survival was 14.6 years. Comparing survival for treated versus age- and sex-matched untreated cohorts, hazard ratio was 0.13 (95% confidence interval, 0.04–0.37; P &lt;0.001) with median follow-up of 5.3 years from time of treatment initiation. There were 21 of 43 deaths in untreated versus 5 of 43 deaths among treated subjects. Treatment increased mean survival by 1.6 years. Conclusions— This study provides a robust untreated disease survival profile that can be used for comparisons now and in the future to assess changes in survival with treatments for Hutchinson-Gilford progeria syndrome. The current comparisons estimating increased survival with protein farnesylation inhibitors provide the first evidence of treatments influencing survival for this fatal disease. Clinical Trial Registration— URL: http://www.clinicaltrials.gov . Unique Indentifiers: NCT00425607, NCT00879034, and NCT00916747.

<h3>Importance</h3> Hutchinson-Gilford progeria syndrome (HGPS) is an extremely rare fatal premature aging disease. There is no approved treatment. <h3>Objective</h3> To evaluate the association of monotherapy using the protein farnesyltransferase inhibitor lonafarnib with mortality rate in children with HGPS. <h3>Design, Setting, and Participants</h3> Cohort study comparing contemporaneous (birth date ≥1991) untreated patients with HGPS matched with treated patients by age, sex, and continent of residency using conditional Cox proportional hazards regression. Treatment cohorts included patients from 2 single-group, single-site clinical trials (ProLon1 [n = 27; completed] and ProLon2 [n = 36; ongoing]). Untreated patients originated from a separate natural history study (n = 103). The cutoff date for patient follow-up was January 1, 2018. <h3>Exposure</h3> Treated patients received oral lonafarnib (150 mg/m²) twice daily. Untreated patients received no clinical trial medications. <h3>Main Outcomes and Measures</h3> The primary outcome was mortality. The primary analysis compared treated patients from the first lonafarnib trial with matched untreated patients. A secondary analysis compared the combined cohorts from both lonafarnib trials with matched untreated patients. <h3>Results</h3> Among untreated and treated patients (n = 258) from 6 continents, 123 (47.7%) were female; 141 (54.7%) had a known genotype, of which 125 (88.7%) were classic (c.1824C&gt;T in<i>LMNA</i>). When identified (n = 73), the primary cause of death was heart failure (79.4%). The median treatment duration was 2.2 years. Median age at start of follow-up was 8.4 (interquartile range [IQR], 4.8-9.5) years in the first trial cohort and 6.5 (IQR, 3.7-9.0) years in the combined cohort. There was 1 death (3.7%) among 27 patients in the first trial group and there were 9 deaths (33.3%) among 27 patients in the matched untreated group. Treatment was associated with a lower mortality rate (hazard ratio, 0.12; 95% CI, 0.01-0.93;<i>P</i> = .04). In the combined cohort, there were 4 deaths (6.3%) among 63 patients in the treated group and 17 deaths (27.0%) among 63 patients in the matched untreated group (hazard ratio, 0.23; 95% CI, 0.06-0.90;<i>P</i> = .04). <h3>Conclusions and Relevance</h3> Among patients with HGPS, lonafarnib monotherapy, compared with no treatment, was associated with a lower mortality rate after 2.2 years of follow-up. Study interpretation is limited by its observational design.

The aim of the study was to review systematically the literature measuring the accuracy of routine UK hospital statistics that classify patients on discharge.A systematic review was carried out of studies comparing routine discharge statistics about an episode of hospital care with the original medical record. Dual quality assessment and extraction was completed for included studies. Qualitative and descriptive analyses were undertaken. Additional comparisons of factors that could potentially introduce systematic variation in coding accuracy were also undertaken.Thirty studies were identified, of which 21 were included in the review. Twelve of these were conducted in England and Wales, and nine in Scotland. The majority assessed the accuracy of a single diagnosis, or selection of diagnoses in a limited range of hospital settings. The median coding accuracy rates were 91 per cent for diagnostic codes and 69.5 per cent for operation or procedure codes in studies in England or Wales; 82 per cent for diagnostic codes and 98 per cent for operation or procedure codes in Scottish studies. There were no significant differences in coding accuracy over time or in the type or rarity of the codes being assessed. Accuracy rates were higher for ICD7 codes (median 96.5 per cent) than for ICD8 (median 87 per cent) or ICD9 (median 77 per cent).Coding accuracy on average is high in the United Kingdom, especially for operations and procedures. However, policy-makers, planners and researchers need to recognize and account for the degree of inaccuracy in routine hospital information statistics. Further research is needed into methods of improving and maintaining coding accuracy.

Hereditary hypouricemia may result from mutations in the renal tubular uric acid transporter URAT1. Whether mutation of other uric acid transporters produces a similar phenotype is unknown. We studied two families who had severe hereditary hypouricemia and did not have a URAT1 defect. We performed a genome-wide homozygosity screen and linkage analysis and identified the candidate gene SLC2A9, which encodes the glucose transporter 9 (GLUT9). Both families had homozygous SLC2A9 mutations: A missense mutation (L75R) in six affected members of one family and a 36-kb deletion, resulting in a truncated protein, in the other. In vitro, the L75R mutation dramatically impaired transport of uric acid. The mean concentration of serum uric acid of seven homozygous individuals was 0.17 +/- 0.2 mg/dl, and all had a fractional excretion of uric acid >150%. Three individuals had nephrolithiasis, and three had a history of exercise-induced acute renal failure. In conclusion, homozygous loss-of-function mutations of GLUT9 cause a total defect of uric acid absorption, leading to severe renal hypouricemia complicated by nephrolithiasis and exercise-induced acute renal failure. In addition to clarifying renal handling of uric acid, our findings may provide a better understanding of the pathophysiology of acute renal failure, nephrolithiasis, hyperuricemia, and gout.

Sphingolipids have essential roles as structural components of cell membranes and in cell signalling, and disruption of their metabolism causes several diseases, with diverse neurological, psychiatric, and metabolic consequences. Increasingly, variants within a few of the genes that encode enzymes involved in sphingolipid metabolism are being associated with complex disease phenotypes. Direct experimental evidence supports a role of specific sphingolipid species in several common complex chronic disease processes including atherosclerotic plaque formation, myocardial infarction (MI), cardiomyopathy, pancreatic beta-cell failure, insulin resistance, and type 2 diabetes mellitus. Therefore, sphingolipids represent novel and important intermediate phenotypes for genetic analysis, yet little is known about the major genetic variants that influence their circulating levels in the general population. We performed a genome-wide association study (GWAS) between 318,237 single-nucleotide polymorphisms (SNPs) and levels of circulating sphingomyelin (SM), dihydrosphingomyelin (Dih-SM), ceramide (Cer), and glucosylceramide (GluCer) single lipid species (33 traits); and 43 matched metabolite ratios measured in 4,400 subjects from five diverse European populations. Associated variants (32) in five genomic regions were identified with genome-wide significant corrected p-values ranging down to 9.08x10(-66). The strongest associations were observed in or near 7 genes functionally involved in ceramide biosynthesis and trafficking: SPTLC3, LASS4, SGPP1, ATP10D, and FADS1-3. Variants in 3 loci (ATP10D, FADS3, and SPTLC3) associate with MI in a series of three German MI studies. An additional 70 variants across 23 candidate genes involved in sphingolipid-metabolizing pathways also demonstrate association (p = 10(-4) or less). Circulating concentrations of several key components in sphingolipid metabolism are thus under strong genetic control, and variants in these loci can be tested for a role in the development of common cardiovascular, metabolic, neurological, and psychiatric diseases.

Central corneal thickness (CCT) is a highly heritable trait, which has been proposed to influence disorders of the anterior segment of the eye. A genome-wide association study (GWAS) of CCT was performed in 2269 individuals from three Croatian and one Scottish population. In the discovery set (1445 individuals), two genome-wide significant associations were identified for single nucleotide polymorphisms rs12447690 (β = 0.23 SD, P = 4.4 × 10(-9)) and rs1536482 (β = 0.22 SD, P = 7.1 × 10(-8)) for which the closest candidate genes (although ≥90 kb away) were zinc finger 469 (ZNF469) on 16q24.2 and collagen 5 alpha 1 (COL5A1) on 9q34.2, respectively. Only the ZNF469 association was confirmed in our replication set (824 individuals, P = 8.0 × 10(-4)) but COL5A1 remained a suggestive association in the combined sample (β = 0.16 SD, P = 1.1 × 10(-6)). Following a larger meta-analysis including recently published CCT GWAS summary data, COL5A1 was genome-wide significant (β = 0.13 SD, P = 5.1 × 10(-8)), together with two additional novel loci. The second new locus (defined by rs1034200) was 5 kb from the AVGR8 gene, encoding a putative transcription factor with typical ZNF and KRAB domains, in chromosomal region 13q12.11 (β = 0.14 SD, P = 3.5 × 10(-9)). The third new locus (rs6496932), on 15q25.3 (β = 0.13, P = 1.4 × 10(-8)), was within a wide linkage disequilibrium block extending into the 5' end of the AKAP13 gene, encoding a scaffold protein concerned with signal transduction from the cell surface. These associations offer mechanistic insights into the regulation of CCT and offer new candidate genes for susceptibility to common disorders in which CCT has been implicated, including primary open-angle glaucoma and keratoconus.

Stephanie London, Martin Tobin and colleagues report meta-analyses of genome-wide association studies for forced vital capacity (FVC), a spirometric measure of pulmonary function that reflects lung volume. They identify six regions newly associated with FVC and demonstrate that candidate genes at these loci are expressed in lung tissue and primary lung cells. Forced vital capacity (FVC), a spirometric measure of pulmonary function, reflects lung volume and is used to diagnose and monitor lung diseases. We performed genome-wide association study meta-analysis of FVC in 52,253 individuals from 26 studies and followed up the top associations in 32,917 additional individuals of European ancestry. We found six new regions associated at genome-wide significance (P < 5 × 10−8) with FVC in or near EFEMP1, BMP6, MIR129-2–HSD17B12, PRDM11, WWOX and KCNJ2. Two loci previously associated with spirometric measures (GSTCD and PTCH1) were related to FVC. Newly implicated regions were followed up in samples from African-American, Korean, Chinese and Hispanic individuals. We detected transcripts for all six newly implicated genes in human lung tissue. The new loci may inform mechanisms involved in lung development and the pathogenesis of restrictive lung disease.

Rabbit hemorrhagic disease virus 2 (RHDV2; Lagovirus GI.2) is a pathogenic calicivirus that affects European rabbits (Oryctolagus cuniculus) and various hare (Lepus) species. GI.2 was first detected in France in 2010 and subsequently caused epidemics in wild and domestic lagomorph populations throughout Europe. In May 2015, GI.2 was detected in Australia. Within 18 months of its initial detection, GI.2 had spread to all Australian states and territories and rapidly became the dominant circulating strain, replacing Rabbit hemorrhagic disease virus (RHDV/GI.1) in mainland Australia. Reconstruction of the evolutionary history of 127 Australian GI.2 isolates revealed that the virus arrived in Australia at least several months before its initial description and likely circulated unnoticed in wild rabbit populations in the east of the continent prior to its detection. GI.2 sequences isolated from five hares clustered with sequences from sympatric rabbit populations sampled contemporaneously, indicating multiple spillover events into hares rather than an adaptation of the Australian GI.2 to a new host. Since the presence of GI.2 in Australia may have wide-ranging consequences for rabbit biocontrol, particularly with the release of the novel biocontrol agent GI.1a/RHDVa-K5 in March 2017, ongoing surveillance is critical to understanding the interactions of the various lagoviruses in Australia and their impact on host populations.IMPORTANCE This study describes the spread and distribution of Rabbit hemorrhagic disease virus 2 (GI.2) in Australia since its first detection in May 2015. Within the first 18 months following its detection, RHDV2 spread from east to west across the continent and became the dominant strain in all mainland states of Australia. This has important implications for pest animal management and for owners of pet and farmed rabbits, as there currently is no effective vaccine available in Australia for GI.2. The closely related RHDV (GI.1) is used to control overabundant wild rabbits, a serious environmental and agricultural pest in this country, and it is currently unclear how the widespread circulation of GI.2 will impact ongoing targeted wild rabbit management operations.

Invasive animals have been linked to the extinctions of native wildlife, and to significant agricultural financial losses or impacts. Current approaches to control invasive species require ongoing resources and management over large geographic scales, and often result in the short-term suppression of populations. New and innovative approaches are warranted. Recently, the RNA guided gene drive system based on CRISPR/Cas9 is being proposed as a potential gene editing tool that could be used by wildlife managers as a non-lethal addition or alternative to help reduce pest animal populations. While regulatory control and social acceptance are crucial issues that must be addressed, there is an opportunity now to identify the knowledge and research gaps that exist for some important invasive species. Here we systematically determine the knowledge gaps for pest species for which gene drives could potentially be applied. We apply a conceptual ecological risk framework within the gene drive context within an Australian environment to identify key requirements for undertaking work on seven exemplar invasive species in Australia. This framework allows an evaluation of the potential research on an invasive species of interest and within a gene drive and risk context. We consider the currently available biological, genetic and ecological information for the house mouse, European red fox, feral cat, European rabbit, cane toad, black rat and European starling to evaluate knowledge gaps and identify candidate species for future research. We discuss these findings in the context of future thematic areas of research worth pursuing in preparation for a more formal assessment of the use of gene drives as a novel strategy for the control of these and other invasive species.

Click to increase image sizeClick to decrease image size Additional informationNotes on contributorsCarol L. ColbeckCarol L. Colbeck is assistant professor and research associate in the Center for the Study of Higher Education at The Pennsylvania State University; Susan E. Campbell is a program evaluator II with the Ohio Legislative Office of Education Oversight; and Stefani A. Bjorklund is a Ph.D. candidate in the Higher Education Program at The Pennsylvania State University.Susan E. CampbellCarol L. Colbeck is assistant professor and research associate in the Center for the Study of Higher Education at The Pennsylvania State University; Susan E. Campbell is a program evaluator II with the Ohio Legislative Office of Education Oversight; and Stefani A. Bjorklund is a Ph.D. candidate in the Higher Education Program at The Pennsylvania State University.Stefani A. BjorklundCarol L. Colbeck is assistant professor and research associate in the Center for the Study of Higher Education at The Pennsylvania State University; Susan E. Campbell is a program evaluator II with the Ohio Legislative Office of Education Oversight; and Stefani A. Bjorklund is a Ph.D. candidate in the Higher Education Program at The Pennsylvania State University.

Hutchinson-Gilford progeria syndrome is a rare and uniformly fatal segmental "premature aging" disease that affects a variety of organ systems. We sought to more clearly define the bone and weight abnormalities in patients with progeria as potential outcome parameters for prospective clinical trials.We collected and analyzed longitudinal medical information, both retrospectively and prospectively, from a total of 41 children with Hutchinson-Gilford progeria syndrome spanning 14 countries, from the Progeria Research Foundation Medical and Research Database at the Brown University Center for Gerontology.In addition to a number of previously well-defined phenotypic findings in children with progeria, this study identified abnormalities in the eruption of secondary incisors lingually and palatally in the mandible and maxilla, respectively. Although bony structures appeared normal in early infancy, clavicular resorption, coxa valga, avascular necrosis of the femoral head, modeling abnormalities of long bones with slender diaphyses, flared metaphyses, and overgrown epiphyses developed. Long bones showed normal cortical thickness centrally and progressive focal demineralization peripherally. The most striking finding identified in the retrospective data set of 35 children was an average weight increase of only 0.44 kg/year, beginning at approximately 24 months of age and persisting through life, with remarkable intrapatient linearity. This rate is >2 SD below normal weight gain for any corresponding age and sharply contrasts with the parabolic growth pattern for normal age- and gender-matched children. This finding was also confirmed prospectively.Our analysis shows evidence of a newly identified abnormal growth pattern for children with Hutchinson-Gilford progeria syndrome. The skeletal and dental findings are suggestive of a developmental dysplasia rather than a classical aging process. The presence of decreased and linear weight gain, maintained in all of the patients after the age of 2 years, provides the ideal parameter on which altered disease status can be assessed in clinical trials.

Abstract A microbially formed soil cover known as desert crust carpets vast regions of land in the arid climates. An accretionary phenomenon in a generally erosional setting, it stabilizes soil and enhances its quality. It is potentially applicable to reclamation of unconsolidated soil in desert regions of the world. This paper reviews the work on microbial crusts, how they develop and contribute to soil quality, and what environmental variables enhance or impede their growth. Microbiological issues relating to the identification, isolation, and culturing of important members of this microbial community are discussed. Some practical problems are considered and suggestions for further research made.

Genes for height have gained interest for decades, but only recently have candidate genes started to be identified. We have performed linkage analysis and genome-wide association for height in approximately 4000 individuals from five European populations. A total of five chromosomal regions showed suggestive linkage and in one of these regions, two SNPs (rs849140 and rs1635852) were associated with height (nominal P = 7.0 x 10(-8) and P = 9.6 x 10(-7), respectively). In total, five SNPs across the genome showed an association with height that reached the threshold of genome-wide significance (nominal P < 1.6 x 10(-7)). The association with height was replicated for two SNPs (rs1635852 and rs849140) using three independent studies (n = 31 077, n=1268 and n = 5746) with overall meta P-values of 9.4 x 10(-10) and 5.3 x 10(-8). These SNPs are located in the JAZF1 gene, which has recently been associated with type II diabetes, prostate and endometrial cancer. JAZF1 is a transcriptional repressor of NR2C2, which results in low IGF1 serum concentrations, perinatal and early postnatal hypoglycemia and growth retardation when knocked out in mice. Both the linkage and association analyses independently identified the JAZF1 region affecting human height. We have demonstrated, through replication in additional independent populations, the consistency of the effect of the JAZF1 SNPs on height. Since this gene also has a key function in the metabolism of growth, JAZF1 represents one of the strongest candidates influencing human height identified so far.

Age-related macular degeneration (AMD) is a leading cause of visual loss in Western populations. Susceptibility is influenced by age, environmental and genetic factors. Known genetic risk loci do not account for all the heritability. We therefore carried out a genome-wide association study of AMD in the UK population with 893 cases of advanced AMD and 2199 controls. This showed an association with the well-established AMD risk loci ARMS2 (age-related maculopathy susceptibility 2)-HTRA1 (HtrA serine peptidase 1) (P =2.7 × 10(-72)), CFH (complement factor H) (P =2.3 × 10(-47)), C2 (complement component 2)-CFB (complement factor B) (P =5.2 × 10(-9)), C3 (complement component 3) (P =2.2 × 10(-3)) and CFI (P =3.6 × 10(-3)) and with more recently reported risk loci at VEGFA (P =1.2 × 10(-3)) and LIPC (hepatic lipase) (P =0.04). Using a replication sample of 1411 advanced AMD cases and 1431 examined controls, we confirmed a novel association between AMD and single-nucleotide polymorphisms on chromosome 6p21.3 at TNXB (tenascin XB)-FKBPL (FK506 binding protein like) [rs12153855/rs9391734; discovery P =4.3 × 10(-7), replication P =3.0 × 10(-4), combined P =1.3 × 10(-9), odds ratio (OR) = 1.4, 95% confidence interval (CI) = 1.3-1.6] and the neighbouring gene NOTCH4 (Notch 4) (rs2071277; discovery P =3.2 × 10(-8), replication P =3.8 × 10(-5), combined P =2.0 × 10(-11), OR = 1.3, 95% CI = 1.2-1.4). These associations remained significant in conditional analyses which included the adjacent C2-CFB locus. TNXB, FKBPL and NOTCH4 are all plausible AMD susceptibility genes, but further research will be needed to identify the causal variants and determine whether any of these genes are involved in the pathogenesis of AMD.

The majority of human proteins are post-translationally modified by covalent addition of one or more complex oligosaccharides (glycans). Alterations in glycosylation processing are associated with numerous diseases and glycans are attracting increasing attention both as disease biomarkers and as targets for novel therapeutic approaches. Using a recently developed high-throughput high-performance liquid chromatography (HPLC) analysis method, we have reported, in a pilot genome-wide association study of 13 glycan features in 2705 individuals from three European populations, that polymorphisms at three loci (FUT8, FUT6/FUT3 and HNF1A) affect plasma levels of N-glycans. Here, we extended the analysis to 33 directly measured and 13 derived glycosylation traits in 3533 individuals and identified three novel gene association (MGAT5, B3GAT1 and SLC9A9) as well as replicated the previous findings using an additional European cohort. MGAT5 (meta-analysis association P-value = 1.80 × 10(-10) for rs1257220) encodes a glycosyltransferase which is known to synthesize the associated glycans. In contrast, neither B3GAT1 (rs7928758, P = 1.66 × 10(-08)) nor SLC9A9 (rs4839604, P = 3.50 × 10(-13)) had previously been associated functionally with glycosylation of plasma proteins. Given the glucuronyl transferase activity of B3GAT1, we were able to show that glucuronic acid is present on antennae of plasma glycoproteins underlying the corresponding HPLC peak. SLC9A9 encodes a proton pump which affects pH in the endosomal compartment and it was recently reported that changes in Golgi pH can impair protein sialylation, giving a possible mechanism for the observed association.

It is a longstanding puzzle why non-coding variants in the complement factor H (CFH) gene are more strongly associated with age-related macular degeneration (AMD) than functional coding variants that directly influence the alternative complement pathway. The situation is complicated by tight genetic associations across the region, including the adjacent CFH-related genes CFHR3 and CFHR1, which may themselves influence the alternative complement pathway and are contained within a common deletion (CNP147) which is associated with protection against AMD. It is unclear whether this association is mediated through a protective effect of low plasma CFHR1 concentrations, high plasma CFH or both. We examined the triangular relationships of CFH/CFHR3/CFHR1 genotype, plasma CFH or CFHR1 concentrations and AMD susceptibility in combined case-control (1256 cases, 1020 controls) and cross-sectional population (n = 1004) studies and carried out genome-wide association studies of plasma CFH and CFHR1 concentrations. A non-coding CFH SNP (rs6677604) and the CNP147 deletion were strongly correlated both with each other and with plasma CFH and CFHR1 concentrations. The plasma CFH-raising rs6677604 allele and raised plasma CFH concentration were each associated with AMD protection. In contrast, the protective association of the CNP147 deletion with AMD was not mediated by low plasma CFHR1, since AMD-free controls showed increased plasma CFHR1 compared with cases, but it may be mediated by the association of CNP147 with raised plasma CFH concentration. The results are most consistent with a regulatory locus within a 32 kb region of the CFH gene, with a major effect on plasma CFH concentration and AMD susceptibility.

Urinary incontinence is common after both radical prostatectomy and transurethral resection of the prostate (TURP). Conservative management includes pelvic floor muscle training with or without biofeedback, electrical stimulation, extra-corporeal magnetic innervation (ExMI), compression devices (penile clamps), lifestyle changes, or a combination of methods.To assess the effects of conservative management for urinary incontinence after prostatectomy.We searched the Cochrane Incontinence Group Specialised Register (searched 24 August 2011), EMBASE (January 1980 to Week 48 2009), CINAHL (January 1982 to 20 November 2009), the reference lists of relevant articles, handsearched conference proceedings and contacted investigators to locate studies.Randomised or quasi-randomised controlled trials evaluating conservative interventions for urinary continence in men after prostatectomy.Two or more review authors assessed the methodological quality of trials and abstracted data. We tried to contact several authors of included studies to obtain extra information.Thirty-seven trials met the inclusion criteria, 33 amongst men after radical prostatectomy, three trials after transurethral resection of the prostate (TURP) and one trial after either operation. The trials included 3399 men, of whom 1937 had an active conservative intervention. There was considerable variation in the interventions, populations and outcome measures. Data were not available for many of the pre-stated outcomes. Men's symptoms improved over time irrespective of management. Adverse effects did not occur or were not reported.There was no evidence from eight trials that pelvic floor muscle training with or without biofeedback was better than control for men who had urinary incontinence after radical prostatectomy (e.g. 57% with urinary incontinence versus 62% in the control group, risk ratio (RR) for incontinence after 12 months 0.85, 95% confidence interval (CI) 0.60 to 1.22) as the confidence intervals were wide, reflecting uncertainty. However, one large multicentre trial of one-to-one therapy showed no difference in any urinary or quality of life outcome measures and had narrower confidence intervals. There was also no evidence of benefit for erectile dysfunction (56% with no erection in the pelvic floor muscle training group versus 55% in the control group after one year, RR 1.01, 95% CI 0.84 to 1.20). Individual small trials provided data to suggest that electrical stimulation, external magnetic innervation or combinations of treatments might be beneficial but the evidence was limited. One large trial demonstrated that there was no benefit for incontinence or erectile dysfunction from a one-to-one pelvic floor muscle training based intervention to men who were incontinent after transurethral resection of the prostate (TURP) (e.g. 65% with urinary incontinence versus 62% in the control group, RR after 12 months 1.05, 95% CI 0.91 to 1.23).In eight trials of conservative treatment of all men after radical prostatectomy aimed at both treatment and prevention, there was an overall benefit from pelvic floor muscle training versus control management in terms of reduction of UI (e.g. 10% with urinary incontinence after one year versus 32% in the control groups, RR for urinary incontinence 0.32, 95% CI 0.20 to 0.51). However, this finding was not supported by other data from pad tests. The findings should be treated with caution, as most trials were of poor to moderate quality and confidence intervals were wide. Men in one trial were more satisfied with one type of external compression device, which had the lowest urine loss, compared to two others or no treatment. The effect of other conservative interventions such as lifestyle changes remains undetermined as no trials involving these interventions were identified.The value of the various approaches to conservative management of postprostatectomy incontinence after radical prostatectomy remains uncertain. It seems unlikely that men benefit from one-to-one pelvic floor muscle training therapy after transurethral resection of the prostate (TURP). Long-term incontinence may be managed by external penile clamp, but there are safety problems.

The TATA binding protein (TBP) plays a central role in eukaryotic and archael transcription initiation.We describe the isolation of a novel 23-kDa human protein that displays 41% identity to TBP and is expressed in most human tissue.Recombinant TBP-related protein (TRP) displayed barely detectable binding to consensus TATA box sequences but bound with slightly higher affinities to nonconsensus TATA sequences.TRP did not substitute for TBP in transcription reactions in vitro.However, addition of TRP potently inhibited basal and activated transcription from multiple promoters in vitro and in vivo.General transcription factors TFIIA and TFIIB bound glutathione S-transferase-TRP in solution but failed to stimulate TRP binding to DNA.Preincubation of TRP with TFIIA inhibited TBP-TFIIA-DNA complex formation and addition of TFIIA overcame TRP-mediated transcription repression.TRP transcriptional repression activity was specifically reduced by mutations in TRP that disrupt the TFIIA binding surface but not by mutations that disrupt the TFIIB or DNA binding surface of TRP.These results suggest that TFIIA is a primary target of TRP transcription inhibition and that TRP may modulate transcription by a novel mechanism involving the partial mimicry of TBP functions.

to examine the relationship between seven predictor variables (recorded on Day 3 of hospital admission) and discharge destination in non-elective medical patients aged 65+ years.prospective cohort.eight centres in six European countries.age, gender, living alone, physical function (three categories based on Barthel Index), cognition (Katzman's orientation-memory-concentration test), main body system affected (based on International Classification of Diseases), number of geriatric giants (GGs) involved in the referral (a GG being a problem with falling, mobility, continence or cognition).discharge destination (by Day 90) in three categories: 'HOMESAME' (return to previous residence), 'INSTIN90' (discharge to alternative residence or still in hospital at 90 days), 'DEADINHO' (death in hospital),in 1,626 patients, discharge destination was HOMESAME in 84.7%, DEADINHO in 8.9% and INSTIN90 in 6.4%. Mean duration of stay was 17.7 days, median 12. Univariate analyses showed a statistically significant relationship between all seven predictor variables and discharge destination. Physical function was the best single predictor with a seven-fold difference in adverse outcome rates between the best and worst categories. On multiple logistic regression, significant predictor variables were as follows. (i) For DEADINHO: physical function, cognition, gender; (ii) for INSTIN90: physical function, living alone, GGs, age, gender. Multiple linear regression identified physical function, GGs and living alone as predictors of loge length of stay.case-mix systems to compare risk-adjusted hospital outcome in older medical patients need to incorporate information about physical function, cognition and presenting problems in addition to diagnosis.

There is increasing evidence for a role of genetic predisposition in the etiology of kidney disease, but linkage scans have been poorly replicated. Here we performed a genome-wide linkage analysis of serum creatinine on 2859 individuals from isolated villages in South Tyrol (Italy), Rucphen (The Netherlands) and Vis Island (Croatia), populations that have been stable and permanently resident in their region. Linkage of serum creatinine levels to loci on chromosomes 7p14, 9p21, 11p15, 15q15-21, 16p13, and 18p11 was successfully replicated in at least one discovery population or in the pooled analysis. A novel locus was found on chromosome 10p11. Linkage to chromosome 22q13, independent of diabetes and hypertension, was detected over a region containing the non-muscle myosin heavy chain type II isoform A (MYH9) gene (LOD score=3.52). In non-diabetic individuals, serum creatinine was associated with this gene in two of the three populations and in meta-analysis (SNP rs11089788, P-value=0.0089). In populations sharing a homogeneous environment and genetic background, heritability of serum creatinine was higher than in outbred populations, with consequent detection of a larger number of loci than reported before. Our finding of a replicated association of serum creatinine with the MYH9 gene, recently linked to pathological renal conditions in African Americans, suggests that this gene may also influence kidney function in healthy Europeans. There is increasing evidence for a role of genetic predisposition in the etiology of kidney disease, but linkage scans have been poorly replicated. Here we performed a genome-wide linkage analysis of serum creatinine on 2859 individuals from isolated villages in South Tyrol (Italy), Rucphen (The Netherlands) and Vis Island (Croatia), populations that have been stable and permanently resident in their region. Linkage of serum creatinine levels to loci on chromosomes 7p14, 9p21, 11p15, 15q15-21, 16p13, and 18p11 was successfully replicated in at least one discovery population or in the pooled analysis. A novel locus was found on chromosome 10p11. Linkage to chromosome 22q13, independent of diabetes and hypertension, was detected over a region containing the non-muscle myosin heavy chain type II isoform A (MYH9) gene (LOD score=3.52). In non-diabetic individuals, serum creatinine was associated with this gene in two of the three populations and in meta-analysis (SNP rs11089788, P-value=0.0089). In populations sharing a homogeneous environment and genetic background, heritability of serum creatinine was higher than in outbred populations, with consequent detection of a larger number of loci than reported before. Our finding of a replicated association of serum creatinine with the MYH9 gene, recently linked to pathological renal conditions in African Americans, suggests that this gene may also influence kidney function in healthy Europeans. It is well established that inherited factors play an important role in the etiology of renal disease.1.Freedman B.I. Satko S.G. Genes and renal disease.Curr Opin Nephrol Hypertens. 2000; 9: 273-277Crossref PubMed Scopus (25) Google Scholar Familial aggregation of both diabetic and IgA nephropathies has been extensively recognized,2.Chow K.M. Wong T.Y. Li P.K. Genetics of common progressive renal disease.Kidney Int Suppl. 2005; 67: S41-S45Abstract Full Text Full Text PDF Google Scholar and several genetic factors contributing to chronic renal failure have been proposed.3.Freedman B.I. Susceptibility genes for hypertension and renal failure.J Am Soc Nephrol. 2003; 14: S192-S194Crossref PubMed Google Scholar The genetic heritability of serum creatinine (SCR), glomerular filtration rate (GFR), and creatinine clearance (CrCl) was shown to be significant and very high, irrespective of ethnicity and concomitant pathologies.4.Puppala S. Arya R. Thameem F. et al.Genotype by diabetes interaction effects on the detection of linkage of glomerular filtration rate to a region on chromosome 2q in Mexican Americans.Diabetes. 2007; 56: 2818-2828Crossref PubMed Scopus (33) Google Scholar, 5.Hunt S.C. Coon H. Hasstedt S.J. et al.Linkage of serum creatinine and glomerular filtration rate to chromosome 2 in Utah pedigrees.Am J Hypertens. 2004; 17: 511-515Crossref PubMed Scopus (33) Google Scholar, 6.Placha G. Poznik G.D. Dunn J. et al.A genome-wide linkage scan for genes controlling variation in renal function estimated by serum cystatin C levels in extended families with type 2 diabetes.Diabetes. 2006; 55: 3358-3365Crossref PubMed Scopus (61) Google Scholar, 7.Turner S.T. Kardia S.L. Mosley T.H. et al.Influence of genomic loci on measures of chronic kidney disease in hypertensive sibships.J Am Soc Nephrol. 2006; 17: 2048-2055Crossref PubMed Scopus (36) Google Scholar, 8.Fox C.S. Yang Q. Cupples L.A. et al.Genomewide linkage analysis to serum creatinine, GFR, and creatinine clearance in a community-based population: the Framingham Heart Study.J Am Soc Nephrol. 2004; 15: 2457-2461Crossref PubMed Scopus (138) Google Scholar, 9.Hunt S.C. Hasstedt S.J. Coon H. et al.Linkage of creatinine clearance to chromosome 10 in Utah pedigrees replicates a locus for end-stage renal disease in humans and renal failure in the fawn-hooded rat.Kidney Int. 2002; 62: 1143-1148Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar, 10.Arar N.H. Voruganti V.S. Nath S.D. et al.A genome-wide search for linkage to chronic kidney disease in a community-based sample: the SAFHS.Nephrol Dial Transplant. 2008; 23: 3184-3191Crossref PubMed Scopus (34) Google Scholar, 11.Mottl A.K. Vupputuri S. Cole S.A. et al.Linkage analysis of glomerular filtration rate in American Indians.Kidney Int. 2008; 74: 1185-1191Abstract Full Text Full Text PDF PubMed Scopus (20) Google Scholar Recently, several genome-wide linkage scans have been carried out in the attempt to detect loci explaining variability of quantitative renal phenotypes in individuals of different ethnicities,4.Puppala S. Arya R. Thameem F. et al.Genotype by diabetes interaction effects on the detection of linkage of glomerular filtration rate to a region on chromosome 2q in Mexican Americans.Diabetes. 2007; 56: 2818-2828Crossref PubMed Scopus (33) Google Scholar, 5.Hunt S.C. Coon H. Hasstedt S.J. et al.Linkage of serum creatinine and glomerular filtration rate to chromosome 2 in Utah pedigrees.Am J Hypertens. 2004; 17: 511-515Crossref PubMed Scopus (33) Google Scholar, 6.Placha G. Poznik G.D. Dunn J. et al.A genome-wide linkage scan for genes controlling variation in renal function estimated by serum cystatin C levels in extended families with type 2 diabetes.Diabetes. 2006; 55: 3358-3365Crossref PubMed Scopus (61) Google Scholar, 7.Turner S.T. Kardia S.L. Mosley T.H. et al.Influence of genomic loci on measures of chronic kidney disease in hypertensive sibships.J Am Soc Nephrol. 2006; 17: 2048-2055Crossref PubMed Scopus (36) Google Scholar, 10.Arar N.H. Voruganti V.S. Nath S.D. et al.A genome-wide search for linkage to chronic kidney disease in a community-based sample: the SAFHS.Nephrol Dial Transplant. 2008; 23: 3184-3191Crossref PubMed Scopus (34) Google Scholar, 11.Mottl A.K. Vupputuri S. Cole S.A. et al.Linkage analysis of glomerular filtration rate in American Indians.Kidney Int. 2008; 74: 1185-1191Abstract Full Text Full Text PDF PubMed Scopus (20) Google Scholar, 12.DeWan A.T. Arnett D.K. Atwood L.D. et al.A genome scan for renal function among hypertensives: the HyperGEN study.Am J Hum Genet. 2001; 68: 136-144Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar, 13.DeWan A.T. Arnett D.K. Miller M.B. et al.Refined mapping of suggestive linkage to renal function in African Americans: the HyperGEN study.Am J Hum Genet. 2002; 71: 204-205Abstract Full Text Full Text PDF PubMed Scopus (15) Google Scholar, 14.Chen G. Adeyemo A.A. Zhou J. et al.A genome-wide search for linkage to renal function phenotypes in West Africans with type 2 diabetes.Am J Kidney Dis. 2007; 49: 394-400Abstract Full Text Full Text PDF PubMed Scopus (46) Google Scholar, 15.Schelling J.R. Abboud H.E. Nicholas S.B. et al.Genome-wide scan for estimated glomerular filtration rate in multi-ethnic diabetic populations: the Family Investigation of Nephropathy and Diabetes (FIND).Diabetes. 2008; 57: 235-243Crossref PubMed Scopus (81) Google Scholar including extended pedigrees of diabetic4.Puppala S. Arya R. Thameem F. et al.Genotype by diabetes interaction effects on the detection of linkage of glomerular filtration rate to a region on chromosome 2q in Mexican Americans.Diabetes. 2007; 56: 2818-2828Crossref PubMed Scopus (33) Google Scholar, 14.Chen G. Adeyemo A.A. Zhou J. et al.A genome-wide search for linkage to renal function phenotypes in West Africans with type 2 diabetes.Am J Kidney Dis. 2007; 49: 394-400Abstract Full Text Full Text PDF PubMed Scopus (46) Google Scholar, 15.Schelling J.R. Abboud H.E. Nicholas S.B. et al.Genome-wide scan for estimated glomerular filtration rate in multi-ethnic diabetic populations: the Family Investigation of Nephropathy and Diabetes (FIND).Diabetes. 2008; 57: 235-243Crossref PubMed Scopus (81) Google Scholar and hypertensive6.Placha G. Poznik G.D. Dunn J. et al.A genome-wide linkage scan for genes controlling variation in renal function estimated by serum cystatin C levels in extended families with type 2 diabetes.Diabetes. 2006; 55: 3358-3365Crossref PubMed Scopus (61) Google Scholar, 7.Turner S.T. Kardia S.L. Mosley T.H. et al.Influence of genomic loci on measures of chronic kidney disease in hypertensive sibships.J Am Soc Nephrol. 2006; 17: 2048-2055Crossref PubMed Scopus (36) Google Scholar, 12.DeWan A.T. Arnett D.K. Atwood L.D. et al.A genome scan for renal function among hypertensives: the HyperGEN study.Am J Hum Genet. 2001; 68: 136-144Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar, 13.DeWan A.T. Arnett D.K. Miller M.B. et al.Refined mapping of suggestive linkage to renal function in African Americans: the HyperGEN study.Am J Hum Genet. 2002; 71: 204-205Abstract Full Text Full Text PDF PubMed Scopus (15) Google Scholar individuals. Some of these studies assessed linkage of GFR or CrCl, estimated by means of common SCR-based equations.16.Stevens L.A. Coresh J. Greene T. et al.Assessing kidney function—measured and estimated glomerular filtration rate.N Engl J Med. 2006; 354: 2473-2483Crossref PubMed Scopus (2300) Google Scholar However, it is worth noting that, when age, sex, and other factors used for estimation of these quantities are included in regression models, there is no meaningful difference between the analysis of estimated GFR (CrCl) and SCR. In this study, we investigated the genetics of SCR in three genetically isolated populations living in Europe and participating in the European Special Population Research Network (EUROSPAN). Isolated populations can play an important role in dissecting complex traits.17.Peltonen L. Palotie A. Lange K. Use of population isolates for mapping complex traits.Nat Rev Genet. 2000; 1: 182-190Crossref PubMed Scopus (310) Google Scholar,18.Shifman S. Darvasi A. The value of isolated populations.Nat Genet. 2001; 28: 309-310Crossref PubMed Scopus (129) Google Scholar One advantage over outbred populations is related to the greater homogeneity of lifestyle and environmental factors, as empirically shown in South Tyrolean isolates.19.Marroni F. Grazio D. Pattaro C. et al.Estimates of genetic and environmental contribution to 43 quantitative traits support sharing of a homogeneous environment in an isolated population from South Tyrol, Italy.Hum Hered. 2008; 65: 175-182Crossref PubMed Scopus (30) Google Scholar Population isolates are characterized by a reduced number of recombination events, typical of pedigrees generated from a small group of founders. Thus, the study of small isolates could facilitate the search for causal loci involved in both monogenic and polygenic disorders.20.Sheffield V.C. Stone E.M. Carmi R. Use of isolated inbred human populations for identification of disease genes.Trends Genet. 1998; 14: 391-396Abstract Full Text Full Text PDF PubMed Scopus (101) Google Scholar, 21.Escamilla M.A. Population isolates: their special value for locating genes for bipolar disorder.Bipolar Disord. 2001; 3: 299-317Crossref PubMed Scopus (39) Google Scholar, 22.Sheffield V.C. Use of isolated populations in the study of a human obesity syndrome, the Bardet–Biedl syndrome.Pediatr Res. 2004; 55: 908-911Crossref PubMed Scopus (20) Google Scholar The study involved 2859 participants from the MICROS study (Italy), the Erasmus Rucphen Family (ERF) study (the Netherlands), and from the island of Vis (Croatia). Characteristics of participants are reported in Table 1. Overall, women were slightly more in number than were men, and Vis participants were 7–9 years older than other participants. Body mass index was higher in ERF and Vis study participants than in the MICROS study participants. ERF had a higher prevalence of ever smokers, diabetes, hypertension, and higher systolic blood pressure. Distribution of renal function indicators suggested a better renal condition in MICROS and ERF than in Vis participants.Table 1Characteristics of study participantsaReported data are mean (s.d.) or N (%), depending on the type of study variable.StudyMICROSERFVisP-valuebUnadjusted homogeneity χ2 test for categorical variables. Kruskal–Wallis test for continuous variables.No. of participants8911388580Determinants of renal function Age (years)47 (16.6)49 (14.6)56 (16.2)<0.00001 Sex (F)487 (54.7)785 (56.6)330 (56.9)0.60232 BMI (kg/m2)26 (4.5)27 (4.6)27 (4.1)<0.00001 Ever smoked (yes)365 (41.1)963 (70.3)294 (50.8)<0.00001 Diabetes (yes)32 (3.7)63 (7.4)35 (6.1)0.00273 Under anti-hypertensive treatment (yes)76 (8.5)284 (34.0)149 (25.8)<0.00001 SBP (mm Hg)133 (20.8)140 (19.8)137 (23.7)<0.00001 DBP (mm Hg)80 (11.2)80 (9.7)80 (11.4)0.69550 MAP (mm Hg)98 (13.5)100 (11.8)99 (14.1)0.00007Renal function Males SCR0.96 (0.175)0.97 (0.181)1.09 (0.22)<0.00001 eGFRceGFR: glomerular filtration rate as estimated with the updated 4-variable modification of diet in renal disease (MDRD) formula.2386 (17.2)86 (17.9)75 (17.5)<0.00001 Females SCR0.80 (0.175)0.79 (0.181)0.91 (0.218)<0.00001 eGFRceGFR: glomerular filtration rate as estimated with the updated 4-variable modification of diet in renal disease (MDRD) formula.2381 (17.2)81 (17.9)67 (17.5)<0.00001BMI, body mass index; DBP, diastolic blood pressure; GFR, glomerular filtration rate; MAP, mean arterial pressure; SCR, serum creatinine; SBP, systolic blood pressure.a Reported data are mean (s.d.) or N (%), depending on the type of study variable.b Unadjusted homogeneity χ2 test for categorical variables. Kruskal–Wallis test for continuous variables.c eGFR: glomerular filtration rate as estimated with the updated 4-variable modification of diet in renal disease (MDRD) formula.23.Levey A.S. Coresh J. Greene T. et al.Using standardized serum creatinine values in the modification of diet in renal disease study equation for estimating glomerular filtration rate.Ann Intern Med. 2006; 145: 247-254Crossref PubMed Scopus (4112) Google Scholar Open table in a new tab BMI, body mass index; DBP, diastolic blood pressure; GFR, glomerular filtration rate; MAP, mean arterial pressure; SCR, serum creatinine; SBP, systolic blood pressure. After pedigree splitting, singletons and individuals with missing information on covariates were eliminated, and finally, 856, 822, and 561 individuals from MICROS, ERF, and Vis, respectively, were retained in the analysis. They were subdivided into 121 (MICROS), 138 (ERF), and 156 (Vis) smaller pedigrees with a median size of 18 individuals (range: 3–27) in MICROS, 19 (12–28) in ERF, and 5.5 (3–25) in Vis. The distribution of relative pairs according to study and genetic relationship is reported in the Supplementary Information File 1. Given a hypothesized total heritability of 40%, the pooled sample showed 76% power to detect a logarithm of odds (LOD) score ≥1.9 for a quantitative trait locus with specific heritability of 18%. The pedigrees from MICROS and ERF cohorts showed substantial power as well. Most likely because of the small size of split pedigrees, the power was much reduced in Vis (an overview of power estimates is given in the Supplementary Information File 2). Download .doc (.06 MB) Help with doc files Supplementary Figures 1–2 Download .doc (.06 MB) Help with doc files Supplementary Table S1 As depicted in Table 2, genetic heritability (h2) of SCR was 0.44 in the pooled sample (P-value=1.09 × 10−31), with a minimum in Vis (h2=0.24, P-value=0.0073) and a maximum in MICROS (h2=0.53, P-value=1.22 × 10–17). The exclusion of diabetics did not affect the estimates. In MICROS, ERF, and in the pooled sample, h2 increased when excluding individuals on anti-hypertensive treatment. In all populations, h2 was higher in non-diabetics not treated for hypertension. The results were similar under the multivariable model (data not shown).Table 2Genetic heritability of serum creatinineMODELAll subjectsSubjects not on anti-hypertensive treatmentNon-diabetics onlySubjects without diabetes and not on anti-hypertensive treatmentStudyh2(s.e.)P-valueh2(s.e.)P-valueh2(s.e.)P-valueh2(s.e.)P-valueERF0.44(0.06)7.00 × 10−170.47(0.09)3.48 × 10−080.41(0.06)4.46 × 10−140.48(0.09)2.97 × 10−08Vis0.24(0.10)0.00730.21(0.18)0.11240.25(0.11)0.00940.31(0.19)0.0505MICROS0.53(0.07)1.22 × 10−170.79(0.09)2.15 × 10−170.52(0.07)1.10 × 10−150.76(0.10)3.32 × 10−16Total0.44(0.04)1.09 × 10−310.53(0.06)1.03 × 10−190.42(0.04)1.74 × 10−270.54(0.06)1.32 × 10−19Estimates (h2), s.e., and P-value are reported by study population (ERF, Vis, MICROS, and pooled), and model (all subjects included; subjects not on anti-hypertensive treatment; non-diabetics only; subjects without diabetes and not on anti-hypertensive treatment).Polygenic models were adjusted for sex, age, and age2. Open table in a new tab Estimates (h2), s.e., and P-value are reported by study population (ERF, Vis, MICROS, and pooled), and model (all subjects included; subjects not on anti-hypertensive treatment; non-diabetics only; subjects without diabetes and not on anti-hypertensive treatment). Polygenic models were adjusted for sex, age, and age2. Linkage regions corresponding to a LOD score ≥1.9 (suggestive linkage) or ≥3.3 (significant linkage) are reported in Table 3. A linkage peak was detected in MICROS on chromosome 7p14 (multivariable LOD score=2.25). Two peaks (at 46 and 49 cM) were observed on chromosome 9p21 in the pooled analysis (Figure 1). The first peak was slightly higher when excluding individuals on anti-hypertensive treatment (multivariable LOD score=2.09). Linkage was more pronounced in the multivariable analysis than in the sex- and age-adjusted one. On chromosome 10p11, suggestive linkage was detected in ERF in individuals not treated for hypertension. Much smaller values were observed when these individuals were included. In the pooled analysis, we detected linkage on chromosome 11p15 in individuals not on anti-hypertensive medication (Figure 1). The signal was similar when diabetics were also removed. On chromosome 15p15, suggestive linkage was observed in the ERF sample under both models, and in all subgroup analyses.Table 3Chromosomal regions corresponding to a LOD score ≥1.9 (suggestive linkageaUnder Lander and Kruglyak's guidelines.28)ChromosomebNCBI Build 36.3, region where the closest marker is located.cMClosest markerscGiven that the three populations had three different maps and the deCODE map was taken as the reference, these are the markers from the deCODE map that are the closest ones to the location of the maximum LOD score.LOD-1 interval (bp)PopulationGroupdALL, no subjects excluded; –AHT, subjects on anti-hypertensive treatment excluded; –DM, subjects with diabetes excluded; –AHT and DM, subjects on anti-hypertensive treatment and diabetics excluded.LOD scoreeNominal and empirical LOD scores estimated under sex- and age-adjusted model (S-A) and multivariable model (M); see the Methods section for details.Previous evidence (linkage)NominalEmpiricalS-AMS-AM7p1457D7S2250-D7S220930,080,312–46,082,813MICROSAll1.952.251.771.86Linkage to the logarithm of SCR in hypertensive sibships of black individuals7.Turner S.T. Kardia S.L. Mosley T.H. et al.Influence of genomic loci on measures of chronic kidney disease in hypertensive sibships.J Am Soc Nephrol. 2006; 17: 2048-2055Crossref PubMed Scopus (36) Google Scholar and to eGFR in the Framingham Heart Study8.Fox C.S. Yang Q. Cupples L.A. et al.Genomewide linkage analysis to serum creatinine, GFR, and creatinine clearance in a community-based population: the Framingham Heart Study.J Am Soc Nephrol. 2004; 15: 2457-2461Crossref PubMed Scopus (138) Google Scholar7p1457D7S2250-D7S220930,080,312–46,082,813MICROS–DM1.982.271.811.939p2146D9S171-D9S167920,347,873–33,163,684Pooled–AHT1.882.291.692.09Overlap with a linkage peak for ESRD in black families enriched for nondiabetic nephropathy24.Freedman B.I. Langefeld C.D. Rich S.S. et al.A genome scan for ESRD in black families enriched for nondiabetic nephropathy.J Am Soc Nephrol. 2004; 15: 2719-2727Crossref PubMed Scopus (42) Google Scholar9p2146D9S171-D9S167920,347,873–33,163,684Pooled–AHT and DM1.842.101.651.949p2149D9S2154-D9S16920,347,873–33,163,684Pooled–AHT and DM1.722.141.551.9810p1161D10S1654-D10S121719,599,223–42,338,905ERF–AHT2.151.982.142.0510p1161D10S1654-D10S121718,880,109–42,338,905ERF–AHT and DM1.911.742.011.6911p1533D11S4106-D11S92815,494,428–23,559,409Pooled–AHT1.911.621.721.45Within the LOD-1 supporting region for linkage to eGFRMDRD in African Americans and in a multiethnic population15.Schelling J.R. Abboud H.E. Nicholas S.B. et al.Genome-wide scan for estimated glomerular filtration rate in multi-ethnic diabetic populations: the Family Investigation of Nephropathy and Diabetes (FIND).Diabetes. 2008; 57: 235-243Crossref PubMed Scopus (81) Google Scholar15q1544D15S537-D15S65934,024,135–53,542,801ERFALL1.941.991.851.86Within the LOD-1 supporting interval for linkage to SCR in West African pedigrees with type II diabetes.14.Chen G. Adeyemo A.A. Zhou J. et al.A genome-wide search for linkage to renal function phenotypes in West Africans with type 2 diabetes.Am J Kidney Dis. 2007; 49: 394-400Abstract Full Text Full Text PDF PubMed Scopus (46) Google Scholar,25.Lichter-Konecki U. Broman K.W. Blau E.B. et al.Genetic and physical mapping of the locus for autosomal dominant renal Fanconi syndrome, on chromosome 15q15.3.Am J Hum Genet. 2001; 68: 264-268Abstract Full Text Full Text PDF PubMed Scopus (31) Google Scholar Linkage to autosomal-dominant renotubular Fanconi syndrome,25.Lichter-Konecki U. Broman K.W. Blau E.B. et al.Genetic and physical mapping of the locus for autosomal dominant renal Fanconi syndrome, on chromosome 15q15.3.Am J Hum Genet. 2001; 68: 264-268Abstract Full Text Full Text PDF PubMed Scopus (31) Google Scholar which can be related to muscle weakness26.Smith R. Lindenbaum R.H. Walton R.J. Hypophosphataemic osteomalacia and Fanconi syndrome of adult onset with dominant inheritance. Possible relationship with diabetes mellitus.Q J Med. 1976; 45: 387-400PubMed Google Scholar15q1544D15S537-D15S65933,171,231–53,542,801ERF–DM1.981.921.821.8215q1544D15S537-D15S65936,794,835–53,542,801ERF–AHT1.881.861.881.9215q2147D15S123-D15S19633,171,231–53,542,801ERF–DM1.811.941.661.8415q2148D15S1028-D15S11936,794,835–53,542,801ERF–AHT2.041.802.031.8715q2148D15S1028-D15S11936,794,835–53,542,801ERF–AHT and DM2.111.932.211.8816p1315D16S423-D16S33924,274,729–8,392,753MICROS–AHT2.442.482.132.00Linkage to early onset ESRD in black families enriched for nondiabetic nephropathy;24.Freedman B.I. Langefeld C.D. Rich S.S. et al.A genome scan for ESRD in black families enriched for nondiabetic nephropathy.J Am Soc Nephrol. 2004; 15: 2719-2727Crossref PubMed Scopus (42) Google Scholar LOD score of 2.0 in a genome-wide scan of UACR in a diabetic, multiethnic population, in majority of Caucasians27.Krolewski A.S. Poznik G.D. Placha G. et al.A genome-wide linkage scan for genes controlling variation in urinary albumin excretion in type II diabetes.Kidney Int. 2006; 69: 129-136Abstract Full Text Full Text PDF PubMed Scopus (97) Google Scholar16p1316D16S423-D16S33924,274,729–10,848,100MICROS–AHT and DM2.352.261.951.8818p1127D18S1163-D18S8434,609,789–9,951,551ERFALL2.462.592.352.42Linkage to GFR estimated by cystatin C in type II diabetic relative pairs (D18S843)6.Placha G. Poznik G.D. Dunn J. et al.A genome-wide linkage scan for genes controlling variation in renal function estimated by serum cystatin C levels in extended families with type 2 diabetes.Diabetes. 2006; 55: 3358-3365Crossref PubMed Scopus (61) Google Scholar18p1127D18S1163-D18S8434,609,789–9,951,551ERF–DM2.692.662.472.5322q1345D22S1045-D22S44534,736,599–47,788,230MICROS–AHT2.493.012.222.42Overlap with the LOD-1 supporting interval for linkage to UACR in a diabetic, multiethnic population, in majority of Caucasians27.Krolewski A.S. Poznik G.D. Placha G. et al.A genome-wide linkage scan for genes controlling variation in urinary albumin excretion in type II diabetes.Kidney Int. 2006; 69: 129-136Abstract Full Text Full Text PDF PubMed Scopus (97) Google Scholar22q1345D22S1045-D22S44532,925,593–47,788,230MICROS–AHT and DM2.763.162.282.6322q1356D22S1171-D22S116837,415,838–47,556,879PooledALL1.921.891.841.8022q1356D22S1171-D22S116841,383,475–47,556,879Pooled–AHT1.891.931.701.7622q1356D22S1171-D22S116841,383,475–47,556,879Pooled–AHT and DM2.172.141.951.9822q1361D22S532-D22S92235,593,050–47,788,230MICROSALL2.683.432.442.8422q1362D22S532-D22S92235,593,050–47,788,230MICROS–DM2.853.522.613.0022q1362D22S532-D22S92234,736,599–47,788,230MICROS–AHT2.552.982.222.4022q1362D22S532-D22S92232,925,593–47,788,230MICROS–AHT and DM2.863.162.372.63eGFR, estimated glomerular filtration rate; ESRD, end-stage renal disease; LOD, logarithm of odds; SCR, serum creatinine; UACR, urinary albumin-to-creatinine ratio.a Under Lander and Kruglyak's guidelines.28.Lander E. Kruglyak L. Genetic dissection of complex traits: guidelines for interpreting and reporting linkage results.Nat Genet. 1995; 11: 241-247Crossref PubMed Scopus (4471) Google Scholarb NCBI Build 36.3, region where the closest marker is located.c Given that the three populations had three different maps and the deCODE map was taken as the reference, these are the markers from the deCODE map that are the closest ones to the location of the maximum LOD score.d ALL, no subjects excluded; –AHT, subjects on anti-hypertensive treatment excluded; –DM, subjects with diabetes excluded; –AHT and DM, subjects on anti-hypertensive treatment and diabetics excluded.e Nominal and empirical LOD scores estimated under sex- and age-adjusted model (S-A) and multivariable model (M); see the Methods section for details. Open table in a new tab eGFR, estimated glomerular filtration rate; ESRD, end-stage renal disease; LOD, logarithm of odds; SCR, serum creatinine; UACR, urinary albumin-to-creatinine ratio. Suggestive linkage to a quantitative trait locus located on chromosome 16p13 was detected in the MICROS sample when excluding individuals on anti-hypertensive treatment, independently of diabetic status (Figure 2). On chromosome 18p11, the sex- and age-adjusted LOD score was 2.46 in the ERF cohort. The signal was higher when excluding diabetics (LOD score=2.69), but disappeared when removing individuals on anti-hypertensive treatment. Similar results were observed with the multivariable analysis. The highest LOD scores were detected on chromosome 22q13 in the MICROS cohort (Figure 2). The first peak, at 45 cM, was observed especially when excluding individuals treated for hypertension and diabetics (multivariable LOD score=3.16). The second peak was observed between 61 and 62 cM. The highest LOD score was obtained when omitting diabetics (LOD score=3.52). Results were consistent and similar across all subgroup analyses, but signals were smaller when adjusting for age and sex only. In the same region, the pooled analysis detected a suggestive linkage peak at 56 cM, where the signal was of similar magnitude regardless of the considered subgroup of individuals and the adjustment applied to the regression model. To assess the consistency of our results with that which was earlier described, we carried out a review of the literature, identifying over 30 articles reporting linkage on SCR, SCR-derived traits, urinary albumin-to-creatinine ratio, and renal disease. For our most prominent findings, the comparison with the literature is reported in Table 3. Apart from the region on chromosome 10p11 and a part of the large region on chromosome 22q13, in all other cases, we confirmed many results reported earlier, and often our findings were supported by more than one earlier study. When extending the comparison to all LOD scores ≥1, the amount of replication was much larger. Of the 54 detected regions, 38 have been reported earlier. This extensive list is provided in the Supplementary Information File 3, together with annotated references to earlier findings. Download .doc (.68 MB)

A 425 million-year-old organically preserved endolithic microfossil, Palaeoconchocelis starmachii Campbell, Kazmierczak, and Golubic (1979), from sedimentary strata of the Upper Silurian of Poland, is identified as the conchocelis phase of a bangiacean rhodophyte.Documentation of the fossil's biological identity is based on several properties shared by the fossil and its living counterpart, conchocelis: their vegetative filaments penetrate carbonate and form a network below the substrate surface (a fossil crinoid columnal and a Recent shell); bulbous swellings occur in series along a vegetative filament; conchosporangial branches develop from the vegetative filaments; conchospores form in the cells of conchosporangial branches; pit connections link cells of the conchocelis. All structural elements of the fossil are equivalent in size, shape, and position within the substrate to modern representatives of the conchocelis phase of Porphyra nereocystis.The extreme plasticity of the biphasic life cycle as well as advantages conferred by the endolithic habit of the conchocelis may be responsible for the unchanged persistence of the bangiacean conchocelis morphology from the Silurian to the present.

The mid-Precambrian microbial fossil Eosynechococcus moorei Hofmann is remarkably similar in morphology to the modern subaerial cyanophyte Gloeothece coerulea Geitler. Eosynechococcus moorei was discovered and described by Hofmann (1976) in cherts of the approximately 1.9 Ga old Belcher Island Formation, Canada, as a member of a microbial assemblage in stromatolites that were interpreted as intertidal. Gloeothece coerulea is a member of a lithophytic microbial assemblage that forms thin crusts on periodically wetted terrestrial rocks. The fossil-to-Recent comparison is based on study of the Eosynechococcus moorei type slides and fresh collections of Gloeothece coerulea from the mountains of Norway. Both the fossil and Recent microorganisms are rod-shaped unicells that have the same cell division pattern, pigmented extracellular envelopes, colony formation, and postmortem degradation sequences. This comparison is based on the following interpretation of the fossil. Ellipsoidal structures with occasional rod-shaped to granular inclusions are interpreted as extracellular envelopes with shrunken cell remnants. This interpretation is consistent with the principles demonstrated in the study of the co-occurring Eoentophysalis belcherensis (Golubic and Hofmann, 1976). The properties listed above were applied in combination at the population level. Gloeothece coerulea is the only plausible morphological counterpart of the mid-Precambrian Eosynechococcus moorei. The occurrence of Eosynechococcus at billion-year intervals in the fossil record, combined with the primitive fine structure of the photosynthetic apparatus (no thylakoid membranes) in its Recent counterpart Gloeothece coerulea (= Gloeobacter violaceus Rippka, Waterbury, and Conen-Bazire) supports the conclusion that this morphotype has been maintained since Precambrian time. The lithophytic assemblage to which Gloeothece coerulea belongs is an example of an extensive prokaryotic vegetational cover on Recent, periodically wetted rocks where higher plants do not grow. Such assemblages must have been common in the past but may not have been preserved as fossils because they occur as crusts on slopes and vertical cliffs with no sediment accumulation. Prokaryotic communities of subaerial habitats are proposed as a model for colonization of terrestrial environments in the Precambrian, long before the evolution of vascular plants.

Fibrin fragment D-dimer, one of several peptides produced when crosslinked fibrin is degraded by plasmin, is the most widely used clinical marker of activated blood coagulation. To identity genetic loci influencing D-dimer levels, we performed the first large-scale, genome-wide association search.A genome-wide investigation of the genomic correlates of plasma D-dimer levels was conducted among 21 052 European-ancestry adults. Plasma levels of D-dimer were measured independently in each of 13 cohorts. Each study analyzed the association between ≈2.6 million genotyped and imputed variants across the 22 autosomal chromosomes and natural-log–transformed D-dimer levels using linear regression in additive genetic models adjusted for age and sex. Among all variants, 74 exceeded the genome-wide significance threshold and marked 3 regions. At 1p22, rs12029080 (P=6.4×10(-52)) was 46.0 kb upstream from F3, coagulation factor III (tissue factor). At 1q24, rs6687813 (P=2.4×10(-14)) was 79.7 kb downstream of F5, coagulation factor V. At 4q32, rs13109457 (P=2.9×10(-18)) was located between 2 fibrinogen genes: 10.4 kb downstream from FGG and 3.0 kb upstream from FGA. Variants were associated with a 0.099-, 0.096-, and 0.061-unit difference, respectively, in natural-log–transformed D-dimer and together accounted for 1.8% of the total variance. When adjusted for nonsynonymous substitutions in F5 and FGA loci known to be associated with D-dimer levels, there was no evidence of an additional association at either locus.Three genes were associated with fibrin D-dimer levels. Of these 3, the F3 association was the strongest, and has not been previously reported.

<h3>BACKGROUND AND PURPOSE:</h3> HGPS is a rare disorder of segmental aging, with early morbidity from cardiovascular and cerebrovascular disease. The goal of this study was to identify the neurovascular features, infarct type, topography, and natural history of stroke in the only neurovascular imaging cohort study of HGPS. <h3>MATERIALS AND METHODS:</h3> We studied 25 children with confirmed diagnoses of HGPS and neuroimaging studies available for review. Relevant clinical information was abstracted from medical records. <h3>RESULTS:</h3> We identified features suggestive of a vasculopathy unique to HGPS, including distinctive intracranial steno-occlusive arterial lesions, basal cistern collateral vessels, and slow compensatory collateral flow over the cerebral convexities. The arterial pathology in the neck consisted of distal vertebral artery stenosis with prominent collateral vessel formation as well as stenosis and calcification of both the cervical internal and common carotid arteries. Radiographic evidence of infarction was found in 60% of patients, of which half were likely clinically silent. Both large- and small-vessel disease was observed, characterized by arterial territorial, white matter, lacunar, and watershed infarcts. <h3>CONCLUSIONS:</h3> We report a unique intracranial and superior cervical arteriopathy in HGPS distinct from other vasculopathies of childhood, such as Moyamoya, and cerebrovascular disease of aging, including atherosclerosis. Arterial features of the mid and lower neck are less distinctive. For the first time, we identified early and clinically silent strokes as a prevalent disease characteristic in HGPS. Longitudinal analysis of stroke incidence and vasculopathy may provide an outcome measure for future treatment interventions for children with HGPS.

We tested for interactions between body mass index (BMI) and common genetic variants affecting serum urate levels, genome-wide, in up to 42569 participants. Both stratified genome-wide association (GWAS) analyses, in lean, overweight and obese individuals, and regression-type analyses in a non BMI-stratified overall sample were performed. The former did not uncover any novel locus with a major main effect, but supported modulation of effects for some known and potentially new urate loci. The latter highlighted a SNP at RBFOX3 reaching genome-wide significant level (effect size 0.014, 95% CI 0.008-0.02, Pinter= 2.6 x 10-8). Two top loci in interaction term analyses, RBFOX3 and ERO1LB-EDARADD, also displayed suggestive differences in main effect size between the lean and obese strata. All top ranking loci for urate effect differences between BMI categories were novel and most had small magnitude but opposite direction effects between strata. They include the locus RBMS1-TANK (men, Pdifflean-overweight= 4.7 x 10-8), a region that has been associated with several obesity related traits, and TSPYL5 (men, Pdifflean-overweight= 9.1 x 10-8), regulating adipocytes-produced estradiol. The top-ranking known urate loci was ABCG2, the strongest known gout risk locus, with an effect halved in obese compared to lean men (Pdifflean-obese= 2 x 10-4). Finally, pathway analysis suggested a role for N-glycan biosynthesis as a prominent urate-associated pathway in the lean stratum. These results illustrate a potentially powerful way to monitor changes occurring in obesogenic environment.

Serum creatinine (S CR) is the most important biomarker for a quick and non-invasive assessment of kidney function in population-based surveys. A substantial proportion of the inter-individual variability in S CR level is explicable by genetic factors.We performed a meta-analysis of genome-wide association studies of S CR undertaken in five population isolates ('discovery cohorts'), all of which are part of the European Special Population Network (EUROSPAN) project. Genes showing the strongest evidence for an association with SCR (candidate loci) were replicated in two additional population-based samples ('replication cohorts').After the discovery meta-analysis, 29 loci were selected for replication. Association between SCR level and polymorphisms in the collagen type XXII alpha 1 (COL22A1) gene, on chromosome 8, and in the synaptotagmin-1 (SYT1) gene, on chromosome 12, were successfully replicated in the replication cohorts (p value = 1.0 x 10(-6) and 1.7 x 10(-4), respectively). Evidence of association was also found for polymorphisms in a locus including the gamma-aminobutyric acid receptor rho-2 (GABRR2) gene and the ubiquitin-conjugating enzyme E2-J1 (UBE2J1) gene (replication p value = 3.6 x 10(-3)). Previously reported findings, associating glomerular filtration rate with SNPs in the uromodulin (UMOD) gene and in the schroom family member 3 (SCHROOM3) gene were also replicated.While confirming earlier results, our study provides new insights in the understanding of the genetic basis of serum creatinine regulatory processes. In particular, the association with the genes SYT1 and GABRR2 corroborate previous findings that highlighted a possible role of the neurotransmitters GABAA receptors in the regulation of the glomerular basement membrane and a possible interaction between GABAA receptors and synaptotagmin-I at the podocyte level.

As major risk‐factors for diabetes and cardiovascular diseases, the genetic contribution to obesity‐related traits has been of interest for decades. Recently, a limited number of common genetic variants, which have replicated in different populations, have been identified. One approach to increase the statistical power in genetic mapping studies is to focus on populations with increased levels of linkage disequilibrium (LD) and reduced genetic diversity. We have performed joint linkage and genome‐wide association analyses for weight and BMI in 3,448 (linkage) and 3,925 (association) partly overlapping healthy individuals from five European populations. A total of four chromosomal regions (two for weight and two for BMI) showed suggestive linkage (lod &gt;2.69) either in one of the populations or in the joint data. At the genome‐wide level (nominal P &lt; 1.6 × 10 −7 , Bonferroni‐adjusted P &lt; 0.05) one single‐nucleotide polymorphism (SNP) (rs12517906) (nominal P = 7.3 × 10 −8 ) was associated with weight, whereas none with BMI. The SNP associated with weight is located close to MGAT1 . The monoacylglycerol acyltransferase (MGAT) enzyme family is known to be involved in dietary fat absorption. There was no overlap between the linkage regions and the associated SNPs. Our results show that genetic effects influencing weight and BMI are shared across diverse European populations, even though some of these populations have experienced recent population bottlenecks and/or been affected by genetic drift. The analysis enabled us to identify a new candidate gene, MGAT1 , associated with weight in women.

Abstract Multi‐strain, host‐disease dynamics describe a system where multiple strains of a pathogen compete for susceptible individuals of a single host. The theoretical properties of these systems have been well studied, but there are few empirical studies in wildlife hosts. We examined the impacts of two novel strains of rabbit haemorrhagic disease virus (RHDV) recently introduced into Australia, one inadvertently (RHDV2) and one deliberately for rabbit biocontrol (RHDV‐K5), by analysing long‐term monitoring data for introduced European rabbits Oryctolagus cuniculus from 18 sites throughout Australia. We examined population‐level impacts using rabbit spotlight counts pre‐ and post‐arrival of the two strains. We also analysed serological data to determine potential interactions among the introduced and existing field strains of RHDV, as well as a pre‐existing benign strain of calicivirus (RCV‐A1). Serological analyses suggested that RHDV2 arrived in Australia during spring 2014 and spread rapidly through the Australian rabbit population within 2 years. Following the establishment of RHDV2, rabbit abundance was reduced by an average of 60%, with impacts most pronounced in South and Western Australia. In contrast, the deliberate release of RHDV‐K5 had little impact on rabbit populations. Although RHDV2 has spread rapidly throughout Australia, our serological analyses do not support the observation that RHDV2 is rapidly replacing existing field strains of RHDV, as was previously reported in Australia and Europe. Nevertheless, RHDV2 has negatively impacted the ability of RHDV and RCV‐A1 to spread within rabbit populations, most likely due to its ability to infect juvenile rabbits, thereby removing them from the pool of susceptible individuals available to be infected by competing strains. Synthesis and applications. The impact of the release of a novel strain of rabbit haemorrhagic disease virus (RHDV‐K5) for rabbit biocontrol in Australia has been suppressed by the emergence of a competing strain, RHDV2. Hence, the success of further releases of similar RHDV strains for rabbit biocontrol appears doubtful. Despite this, RHDV2 has suppressed rabbit abundances by an average of 60%, with impacts most pronounced in South and Western Australia. Whether the incursion of RHDV2 leads to the competitive exclusion of other endemic RHDV strains remains to be resolved. However, the existence of partial cross‐immunity could allow some level of coexistence between RHDV2 and RHDV strains, at least in the medium term.

Following the arrival of rabbit haemorrhagic disease virus 2 (RHDV2) in Australia, average rabbit population abundances were reduced by 60% between 2014 and 2018 based on monitoring data acquired from 18 sites across Australia. During this period, as the seropositivity to RHDV2 increased, concurrent decreases were observed in the seroprevalence of both the previously circulating RHDV1 and RCVA, a benign endemic rabbit calicivirus. However, the detection of substantial RHDV1 seropositivity in juvenile rabbits suggested that infections were continuing to occur, ruling out the rapid extinction of this variant. Here we investigate whether the co-circulation of two pathogenic RHDV variants was sustained after 2018 and whether the initially observed impact on rabbit abundance was still maintained. We monitored rabbit abundance and seropositivity to RHDV2, RHDV1 and RCVA at six of the initial eighteen sites until the summer of 2022. We observed sustained suppression of rabbit abundance at five of the six sites, with the average population reduction across all six sites being 64%. Across all sites, average RHDV2 seroprevalence remained high, reaching 60-70% in adult rabbits and 30-40% in juvenile rabbits. In contrast, average RHDV1 seroprevalence declined to <3% in adult rabbits and 5-6% in juvenile rabbits. Although seropositivity continued to be detected in a low number of juvenile rabbits, it is unlikely that RHDV1 strains now play a major role in the regulation of rabbit abundance. In contrast, RCVA seropositivity appears to be reaching an equilibrium with that of RHDV2, with RCVA seroprevalence in the preceding quarter having a strong negative effect on RHDV2 seroprevalence and vice versa, suggesting ongoing co-circulation of these variants. These findings highlight the complex interactions between different calicivirus variants in free-living rabbit populations and demonstrate the changes in interactions over the course of the RHDV2 epizootic as it has moved towards endemicity. While it is encouraging from an Australian perspective to see sustained suppression of rabbit populations in the eight years following the arrival of RHDV2, it is likely that rabbit populations will eventually recover, as has been observed with previous rabbit pathogens.

Rhegmatogenous retinal detachment (RRD) is an important cause of vision loss and can potentially lead to blindness. The underlying pathogenesis is complex and incompletely understood. We applied a two-stage genetic association discovery phase followed by a replication phase in a combined total of 2833 RRD cases and 7871 controls. The discovery phase involved a genome-wide association scan of 867 affected individuals and 1953 controls from Scotland, followed by genotyping and testing 4347 highest ranking or candidate single nucleotide polymorphisms (SNPs) in independent sets of cases (1000) and controls (2912) of Dutch and British origin. None of the SNPs selected reached a Bonferroni-corrected threshold for significance (P < 1.27 × 10(-7)). The strongest association, for rs12960119 (P = 1.58 × 10(-7)) located within an intron of the SS18 gene. Further testing was carried out in independent case-control series from London (846 cases) and Croatia (120 cases). The combined meta-analysis identified one association reaching genome-wide significance for rs267738 (OR = 1.29, P = 2.11 × 10(-8)), a missense coding SNP and eQTL for CERS2 encoding the protein ceramide synthase 2. Several of the top signals showing suggestive significance in the combined meta-analysis encompassed genes with a documented role in cell adhesion or migration, including SS18, TIAM1, TSTA3 and LDB2, which warrant further investigation. This first genetic association study of RRD supports a polygenic component underlying RRD risk since 27.4% of the underlying RRD liability could be explained by the collective additive effects of the genotyped SNP from the discovery genome-wide scan.

Hutchinson-Gilford progeria syndrome (HGPS) is a fatal sporadic autosomal dominant premature ageing disease caused by single base mutations that optimise a cryptic splice site within exon 11 of the LMNA gene. The resultant disease-causing protein, progerin, acts as a dominant negative. Disease severity relies partly on progerin levels.We report a novel form of somatic mosaicism, where a child possessed two cell populations with different HGPS disease-producing mutations of the same nucleotide-one producing severe HGPS and one mild HGPS. The proband possessed an intermediate phenotype. The mosaicism was initially discovered when Sanger sequencing showed a c.1968+2T>A mutation in blood DNA and a c.1968+2T>C in DNA from cultured fibroblasts. Deep sequencing of DNA from the proband's blood revealed 4.7% c.1968+2T>C mutation, and 41.3% c.1968+2T>A mutation.We hypothesise that the germline mutation was c.1968+2T>A, but a rescue event occurred during early development, where the somatic mutation from A to C at 1968+2 provided a selective advantage. This type of mosaicism where a partial phenotypic rescue event results from a second but milder disease-causing mutation in the same nucleotide has not been previously characterised for any disease.

Abstract Late-onset retinal degeneration (L-ORD) is a rare autosomal dominant retinal dystrophy, characterised by extensive sub-retinal pigment epithelium (RPE) deposits, RPE atrophy, choroidal neovascularisation and photoreceptor cell death associated with severe visual loss. L-ORD shows striking phenotypic similarities to age-related macular degeneration (AMD), a common and genetically complex disorder, which can lead to misdiagnosis in the early stages. To date, a single missense mutation (S163R) in the C1QTNF5 gene, encoding C1q And Tumor Necrosis Factor Related Protein 5 (C1QTNF5) has been shown to cause L-ORD in a subset of affected families. Here, we describe the identification and characterisation of three novel pathogenic mutations in C1QTNF5 in order to elucidate disease mechanisms . In silico and in vitro characterisation show that these mutations perturb protein folding, assembly or polarity of secretion of C1QTNF5 and, importantly, all appear to destabilise the wildtype protein in co-transfection experiments in a human RPE cell line. This suggests that the heterozygous mutations in L-ORD show a dominant negative, rather than a haploinsufficient, disease mechanism. The function of C1QTNF5 remains unclear but this new insight into the pathogenetic basis of L-ORD has implications for future therapeutic strategies such as gene augmentation therapy.

Time to fall asleep (sleep latency) is a major determinant of sleep quality. Chronic, long sleep latency is a major characteristic of sleep-onset insomnia and/or delayed sleep phase syndrome. In this study we aimed to discover common polymorphisms that contribute to the genetics of sleep latency. We performed a meta-analysis of genome-wide association studies (GWAS) including 2 572 737 single nucleotide polymorphisms (SNPs) established in seven European cohorts including 4242 individuals. We found a cluster of three highly correlated variants (rs9900428, rs9907432 and rs7211029) in the RNA-binding protein fox-1 homolog 3 gene (RBFOX3) associated with sleep latency (P-values=5.77 × 10(-08), 6.59 × 10(-)(08) and 9.17 × 10(-)(08)). These SNPs were replicated in up to 12 independent populations including 30 377 individuals (P-values=1.5 × 10(-)(02), 7.0 × 10(-)(03) and 2.5 × 10(-)(03); combined meta-analysis P-values=5.5 × 10(-07), 5.4 × 10(-07) and 1.0 × 10(-07)). A functional prediction of RBFOX3 based on co-expression with other genes shows that this gene is predominantly expressed in brain (P-value=1.4 × 10(-316)) and the central nervous system (P-value=7.5 × 10(-)(321)). The predicted function of RBFOX3 based on co-expression analysis with other genes shows that this gene is significantly involved in the release cycle of neurotransmitters including gamma-aminobutyric acid and various monoamines (P-values<2.9 × 10(-11)) that are crucial in triggering the onset of sleep. To conclude, in this first large-scale GWAS of sleep latency we report a novel association of variants in RBFOX3 gene. Further, a functional prediction of RBFOX3 supports the involvement of RBFOX3 with sleep latency.

Retinal detachment (RD) is a serious and common condition, but genetic studies to date have been hampered by the small size of the assembled cohorts. In the UK Biobank data set, where RD was ascertained by self-report or hospital records, genetic correlations between RD and high myopia or cataract operation were, respectively, 0.46 (SE = 0.08) and 0.44 (SE = 0.07). These correlations are consistent with known epidemiological associations. Through meta-analysis of genome-wide association studies using UK Biobank RD cases (N = 3 977) and two cohorts, each comprising ~1 000 clinically ascertained rhegmatogenous RD patients, we uncovered 11 genome-wide significant association signals. These are near or within ZC3H11B, BMP3, COL22A1, DLG5, PLCE1, EFEMP2, TYR, FAT3, TRIM29, COL2A1 and LOXL1. Replication in the 23andMe data set, where RD is self-reported by participants, firmly establishes six RD risk loci: FAT3, COL22A1, TYR, BMP3, ZC3H11B and PLCE1. Based on the genetic associations with eye traits described to date, the first two specifically impact risk of a RD, whereas the last four point to shared aetiologies with macular condition, myopia and glaucoma. Fine-mapping prioritized the lead common missense variant (TYR S192Y) as causal variant at the TYR locus and a small set of credible causal variants at the FAT3 locus. The larger study size presented here, enabled by resources linked to health records or self-report, provides novel insights into RD aetiology and underlying pathological pathways.

Abstract Background Numerous studies in veterinary species have recently linked vitamin D status with nonskeletal health disorders. Previous studies have indicated that dogs cannot produce endogenous vitamin D via cutaneous production and rely solely on dietary intake of vitamin D. The seasonal variation of vitamin D seen in humans due to changes in ultraviolet (UV) exposure, therefore, is unlikely to be replicated in these animals. Objectives The objective of this study was to investigate the natural variation in 25‐hydroxyvitamin‐D concentrations in dogs subject to seasonal UV exposure. Methods This longitudinal study followed 18 healthy dogs fed a standardized diet over 1 year, with blood samples obtained monthly. Two key vitamin D metabolites, 25‐hydroxyvitamin‐D 2 and 25‐hydroxyvitamin‐D 3 , were assessed by liquid chromatography‐tandem mass spectrometry in serum samples. Various other biochemical parameters were also measured. Seasonality was assessed using cosinor statistical analysis. Results Although the dogs were subject to seasonally varying UV radiation, 25‐hydroxyvitamin‐D and related biomarkers (including calcium and parathyroid hormone) remained stable over time and did not follow a seasonal pattern. 25‐hydroxyvitamin‐D was not positively correlated with exposure to UV radiation. Nonetheless, variation in 25‐hydroxyvitamin‐D concentrations between individual dogs was detected. Conclusions Given the standardization of diet, we concluded that the seasonal stability of 25‐hydroxyvitamin‐D concentration (vitamin D status) was likely a direct result of lack of cutaneous vitamin D production in this species and highlights the importance of dietary intake. The variation in 25‐hydroxyvitamin‐D concentration between animals warrants further investigation.

The characterization of c-Myc target genes, such as rcl and lactate dehydrogenase A (LDH-A), is critical for understanding the mechanisms of c-Myc-induced cell transformation and tumorigenesis. We have previously demonstrated that Rcl induces anchorage-independent growth in Ratla fibroblasts and that LDH-A is required for cell transformation by c-Myc. In this study, we report that Rcl and LDH-A act synergistically to induce anchorage-independent growth. Cells expressing both Rcl and LDH-A form tumors after s.c. injection into nude mice, although neither Rcl or LDH-A overexpression alone induces tumorigenesis. The inability of Rcl and LDH-A to fully recapitulate c-Myc activity, however, indicates that other c-Myc target genes participate in tumorigenesis. In addition, cells that coexpress Rcl and vascular endothelial growth factor are more comparable with c-Myc overexpressing cells in their ability to form tumors in nude mice. These findings confirm Rcl and LDH-A as critical components of the cell transformation program induced by c-Myc and suggest that Rcl is tumorigenic in cells that are provided with a permissive metabolic milieu.

The mosaic structure and molecular evolution of the leukotoxin operon (lktCABD) was investigated by nucleotide sequence comparison of the lktC, lktB, and lktD genes in 23 Mannheimia (Pasteurella) haemolytica, 6 Mannheimia glucosida, and 4 Pasteurella trehalosi strains. Sequence variation in the lktA gene has been described previously (R. L. Davies et al., J. Bacteriol. 183:1394-1404, 2001). The leukotoxin operon of M. haemolytica has a complex mosaic structure and has been derived by extensive inter- and intraspecies horizontal DNA transfer and intragenic recombination events. However, the pattern of recombination varies throughout the operon and among the different evolutionary lineages of M. haemolytica. The lktA and lktB genes have the most complex mosaic structures with segments derived from up to four different sources, including M. glucosida and P. trehalosi. In contrast, the lktD gene is highly conserved in M. haemolytica. The lktC, lktA, and lktB genes of strains representing the major ovine lineages contain recombinant segments derived from bovine or bovine-like serotype A2 strains. These findings support the previous conclusion that host switching of bovine A2 strains from cattle to sheep has played a major role in the evolution of the leukotoxin operon in ovine strains of M. haemolytica. Homologous segments of donor and recipient alleles are identical, or nearly identical, indicating that the recombinational exchanges occurred relatively recent in evolutionary terms. The 5' and 3' ends of the operon are highly conserved in M. haemolytica, which suggests that multiple horizontal exchanges of the complete operon have occurred by a common mechanism such as transduction. Although the lktA and lktB genes both have complex mosaic structures and high nucleotide substitution rates, the amino acid diversity of LktB is significantly lower than that of LktA due to a higher degree of evolutionary constraint against amino acid replacement. The recombinational exchanges within the leukotoxin operon have had greatest effect on LktA and probably provide an adaptive advantage against the host antibody response by generating novel antigenic variation at surface-exposed sites.

Stress causes symptom exacerbation in functional disorders of the urinary bladder. However, the potential mediators and underlying mechanisms of stress effects on micturition reflex function are unknown. We have characterized PACAP (Adcyap1) and PAC1 receptor (Adcyap1r1) signaling in stress-induced urinary bladder dysfunction in mice. We determined PACAP and PAC1 transcripts and protein expressions in the urinary bladder and lumbosacral dorsal root ganglia (DRG) and spinal cord in repeated variate stress (RVS) or control mouse (handling only) groups. RVS in mice significantly (p ≤ 0.01) increased serum corticosterone and urinary bladder NGF content and decreased weight gain. PACAP and PAC1 mRNA and protein were differentially regulated in lower urinary tract tissues with changes observed in lumbosacral DRG and spinal cord but not in urinary bladder. RVS exposure in mice significantly (p ≤ 0.01) increased (2.5-fold) voiding frequency as determined using conscious cystometry. Intrabladder administration of the PAC1 receptor antagonist, PACAP(6-38) (300 nM), significantly (p ≤ 0.01) increased infused volume (1.5-2.7-fold) to elicit a micturition event and increased the intercontraction interval (i.e., decreased voiding frequency) in mice exposed to RVS and in control mice, but changes were smaller in magnitude in control mice. We also evaluated the effect of PAC1 blockade at the level of the urinary bladder on pelvic sensitivity in RVS or control mouse groups using von Frey filament testing. Intrabladder administration of PACAP(6-38) (300 nM) significantly (p ≤ 0.01) reduced pelvic sensitivity following RVS. PACAP/receptor signaling in the CNS and PNS contributes to increased voiding frequency and pelvic sensitivity following RVS and may represent a potential target for therapeutic intervention.

Summary DNA base damage is a major source of oncogenic mutations 1 . Such damage can produce strand-phased mutation patterns and multiallelic variation through the process of lesion segregation 2 . Here, we exploited these properties to reveal how strand-asymmetric processes, such as replication and transcription, shape DNA damage and repair. Despite distinct mechanisms of leading and lagging strand replication 3,4 , we observe identical fidelity and damage tolerance for both strands. For small DNA adducts, our results support a model in which the same translesion polymerase is recruited on-the-fly to both replication strands, starkly contrasting the strand asymmetric tolerance of bulky adducts 5 . We find that DNA damage tolerance is also common during transcription, where RNA-polymerases frequently bypass lesions without triggering repair. At multiple genomic scales, we show the pattern of DNA damage induced mutations is largely shaped by the influence of DNA accessibility on repair efficiency, rather than gradients of DNA damage. Finally, we reveal specific genomic conditions that can corrupt the fidelity of nucleotide excision repair and actively drive oncogenic mutagenesis. These results provide insight into how strand-asymmetric mechanisms underlie the formation, tolerance, and repair of DNA damage, thereby shaping cancer genome evolution.

Symptom exacerbation due to stress is prevalent in many disease states, including functional disorders of the urinary bladder (e.g., overactive bladder (OAB), interstitial cystitis/bladder pain syndrome (IC/BPS)); however, the mechanisms underlying the effects of stress on micturition reflex function are unclear. In this study we designed and evaluated a stress-induced symptom exacerbation (SISE) mouse model that demonstrates increased urinary frequency and somatic (pelvic and hindpaw) sensitivity. Cyclophosphamide (CYP) (35 mg/kg; i.p., every 48 hours for a total of 4 doses) or 7 days of repeated variate stress (RVS) did not alter urinary bladder function or somatic sensitivity; however, both CYP alone and RVS alone significantly (p ≤ 0.01) decreased weight gain and increased serum corticosterone. CYP treatment when combined with RVS for 7 days (CYP+RVS) significantly (p ≤ 0.01) increased serum corticosterone, urinary frequency and somatic sensitivity and decreased weight gain. CYP+RVS exposure in mice significantly (p ≤ 0.01) increased (2.6-fold) voiding frequency as we determined using conscious, open-outlet cystometry. CYP+RVS significantly (p ≤ 0.05) increased baseline, threshold, and peak micturition pressures. We also evaluated the expression of NGF, BDNF, CXC chemokines and IL-6 in urinary bladder in CYP alone, RVS alone and CYP+RVS mouse cohorts. Although all treatments or exposures increased urinary bladder NGF, BDNF, CXC and IL-6 content, CYP+RVS produced the largest increase in all inflammatory mediators examined. These results demonstrated that CYP alone or RVS alone creates a change in the inflammatory environment of the urinary bladder but does not result in a change in bladder function or somatic sensitivity until CYP is combined with RVS (CYP+RVS). The SISE model of CYP+RVS will be useful to develop testable hypotheses addressing underlying mechanisms where psychological stress exacerbates symptoms in functional bladder disorders leading to identification of targets and potential treatments.

Australia has multiple lagoviruses with differing pathogenicity. The circulation of these viruses was traditionally determined through opportunistic sampling events. In the lead up to the nationwide release of RHDVa-K5 (GI.1aP-GI.1a) in 2017, an existing citizen science program, RabbitScan, was augmented to allow members of the public to submit samples collected from dead leporids for lagovirus testing. This study describes the information obtained from the increased number of leporid samples received between 2015 and 2022 and focuses on the recent epidemiological interactions and evolutionary trajectory of circulating lagoviruses in Australia between October 2020 and December 2022. A total of 2771 samples were tested from January 2015 to December 2022, of which 1643 were lagovirus-positive. Notable changes in the distribution of lagovirus variants were observed, predominantly in Western Australia, where RHDV2-4c (GI.4cP-GI.2) was detected again in 2021 after initially being reported to be present in 2018. Interestingly, we found evidence that the deliberately released RHDVa-K5 was able to establish and circulate in wild rabbit populations in WA. Overall, the incorporation of citizen science approaches proved to be a cost-efficient method to increase the sampling area and enable an in-depth analysis of lagovirus distribution, genetic diversity, and interactions. The maintenance of such programs is essential to enable continued investigations of the critical parameters affecting the biocontrol of feral rabbit populations in Australia, as well as to enable the detection of any potential future incursions.

The King's Fund and British Association of Parenteral and Enteral Nutrition recommend that all hospital patients should have height and weight recorded, to detect the need for nutritional support. Systematic review evidence also suggests that protein and energy supplementation of adults in hospital with a wide range of conditions improves outcome.To assess the recording of weight and height in hospitals.Survey (random sample).As part of a survey on the provision of deep venous thrombosis prophylaxis, we collected information on height and weight recording from medical and nursing notes. We randomly selected five medical, five surgical, five orthopaedic, and five obstetrics and gynaecology directorates from across Scotland. Six hundred case notes were requested, and 88% were available for data extraction. Some 67% of hospital episodes provided information about weight, and 41% on both height and weight. General medicine directorates had the lowest recording of weight, and in medical and surgical directorates, both weight and height were rarely recorded in comparison with the other two directorates (p<0.001).Our survey suggests that recommendations to assess nutritional risk are not being followed, and that many patients at risk of malnutrition are not being detected or treated.

Microbial endoliths occur commonly in tests of planktonic and benthic foraminifera, as in other types of skeletal and non-skeletal carbonate substrates in marine sediments. The modem sediments considered in this study were deposited in depths ranging from 200 to more than 4,000 meters. Microbial boring is a strictly benthic activity which afflicts carbonate particles after they have settled at the sediment/water interface. The trace-fossil record of microbial boring is thus a benthic overprint on the carbonate sedimentary record, regardless of its composition. A new morphologically characteristic endolith was found only in deep water sediments (between 2,000 and 4,000 m depths). Other modem heterotrophic endoliths appear to be less restricted in their depth distribution. There is no apparent correlation of occurrence between particular endolithic and foraminiferal taxa, indicating that the relationship is not a host-parasite one, and that any nutrient requirement by the heterotrophic endolith taxa studied is not specific for the organic product of any particular foraminiferal species. Some endolith species and assemblages of species are persistent through geological time (Cretaceous-Recent, Oligocene-Recent). As both microbial endoliths and their host substrates, the benthic foraminifera, have independent depth distributions, their co-occurrence provides a basis for a more refined cross-reference for paleobathymetric in-

To identify case-mix variables measured shortly after admission to be included in a patient classification system (ACMEplus) that best explains hospital outcome for older people in different health care systems.Observational prospective cohort study collecting patient factors (sociodemographics, functional, mental, clinical, administrative and perceived health) at different time assessments.Multicentre study involving eight hospitals in six European countries (United Kingdom, Spain, Italy, Finland, Greece and Poland). It included consecutive patients aged 65 years or older admitted to hospital for acute medical problems.discharge status, hospital readmission, mortality and length of stay.Of the 1667 included patients (mean age = 78.1 years; male gender = 43.5%) two-third had at least one 'Geriatric Giant' (immobility, confusion, incontinence or falls) on admission or shortly after. The most frequently affected system was cardiovascular (29.2%) and 31% of patients declared poor or very poor health. Mean length of stay was 17.9 days, 79% of patients were discharged to their usual residence; in-hospital and 1-month follow up mortality were 7.4% and 11.6%, respectively. Physical function explained the highest variation (between 8% and 21%), followed by cognitive status and number of Geriatric Giants, for almost all outcomes except readmission.Factors other than diagnosis (physical function, cognition and presenting problems) are important in predicting key outcomes of acute hospital care for older people and are consistent across countries. Their inclusion in a standardized system of measurement may be a way of improving quality and equity of medical care in older people.

AimTo identify genetic variants underlying six anthropometric traits: body height, body weight, body mass index, brachial circumference, waist circumference, and hip circumference, using a genome-wide association study.MethodsThe study was carried out in the isolated population of the island of Korčula, Croatia, with 898 adult examinees who participated in the larger DNA-based genetic epidemiological study in 2007. Anthropometric measurements followed standard internationally accepted procedures. Examinees were genotyped using HumanHap 370CNV chip by Illumina, with a genome-wide scan containing 316 730 single nucleotide polymorphisms (SNP).ResultsA total of 11 SNPs were associated with the investigated traits at the level of P < 10−5, with one SNP (rs7792939 in gene zinc finger protein 498, ZNF498) associated with body weight, hip circumference, and brachial circumference (P = 3.59-5.73 × 10−6), and another one (rs157350 in gene delta-sarcoglycan, SGCD) with both brachial and hip circumference (P = 3.70-6.08 × 10−6). Variants in CRIM1, a gene regulating delivery of bone morphogenetic proteins to the cell surface, and ITGA1, involved in the regulation of mesenchymal stem cell proliferation and cartilage production, were also associated with brachial circumference (P = 7.82 and 9.68 × 10−6, respectively) and represent interesting functional candidates. Other associations involved those between genes SEZ6L2 and MAX and waist circumference, XTP6 and brachial circumference, and AMPA1/GRIA1 and height.ConclusionAlthough the study was underpowered for the reported associations to reach formal threshold of genome-wide significance under the assumption of independent multiple testing, the consistency of association between the 2 variants and a set of anthropometric traits makes CRIM1 and ITGA1 highly interesting for further replication and functional follow-up. Increased linkage disequilibrium between the used markers in an isolated population makes the formal significance threshold overly stringent, and changed allele frequencies in isolate population may contribute to identifying variants that would not be easily identified in large outbred populations.

To assess the value of conducting a glaucoma screening randomized controlled trial in the UK.Decision model based economic evaluation and value of information analysis. Model derived from a previous health technology assessment. Model updated in terms of structure and parameter estimates with data from surveys, interviews with members of the public and health care providers and routine sources.On average, across a range of ages of initiating screening (40-60 years), glaucoma prevalence (1-5%), screening uptake (30-100%), and the performance of current case finding, screening was not cost-effective at a £30,000 threshold per quality adjusted life year (QALY) from the perspective of the National Health Service (NHS). The societal value of removing all uncertainty around glaucoma screening is £107 million at a threshold of £20,000 per QALY. For informing policy decisions on glaucoma screening, reducing uncertainty surrounding the NHS and personal social care cost of sight impairment (£74 million) was of most value, followed by reducing uncertainty in test performance (£14 million) and uptake of either screening or current eye care (£8 million each).A glaucoma screening trial in the UK is unlikely to be the best use of research resources. Further research to quantify the costs of sight impairment falling on the NHS and personal social services is a priority. Further development of glaucoma tests and research into strategies to promote the uptake of screening or current eye care such as through the use of a behavioural intervention would be worthwhile.

HGPS is a rare syndrome of segmental premature aging. Our goal was to expand the scope of structural bone and soft-tissue craniofacial abnormalities in HGPS through CT or MR imaging. Using The Progeria Research Foundation Medical and Research Database, 98 imaging studies on 25 patients, birth to 14.1 years of age, were comprehensively reviewed. Eight newly identified abnormalities involving the calvaria, skull base, and soft tissues of the face and orbits were present with prevalences between 43% and 100%. These included J-shaped sellas, a mottled appearance and increased vascular markings of the calvaria, abnormally configured mandibular condyles, hypoplastic articular eminences, small zygomatic arches, prominent parotid glands, and optic nerve kinking. This expanded craniofacial characterization helps link disease features and improves our ability to evaluate how underlying genetic and cellular abnormalities culminate in a disease phenotype.

Biological invasions are not only a major threat to biodiversity, they also have major impacts on local economies and agricultural production systems. Once established, the connection of local populations into metapopulation networks facilitates dispersal at landscape scales, generating spatial dynamics that can impact the outcome of pest-management actions. Much planning goes into landscape-scale invasive species management. However, effective management requires knowledge on the interplay between metapopulation network topology and management actions. We address this knowledge gap using simulation models to explore the effectiveness of two common management strategies, applied across different extents and according to different rules for selecting target localities in metapopulations with different network topologies. These management actions are: (i) general population reduction, and (ii) reduction of an obligate resource. The reduction of an obligate resource was generally more efficient than population reduction for depleting populations at landscape scales. However, the way in which local populations are selected for management is important when the topology of the metapopulation is heterogeneous in terms of the distribution of connections among local populations. We tested these broad findings using real-world scenarios of European rabbits (Oryctolagus cuniculus) infesting agricultural landscapes in Western Australia. Although management strategies targeting central populations were more effective in simulated heterogeneous metapopulation structures, no difference was observed in real-world metapopulation structures that are highly homogeneous. In large metapopulations with high proximity and connectivity of neighbouring populations, different spatial management strategies yield similar outcomes. Directly considering spatial attributes in pest-management actions will be most important for metapopulation networks with heterogeneously distributed links. Our modelling framework provides a simple approach for identifying the best possible management strategy for invasive species based on metapopulation structure and control capacity. This information can be used by managers trying to devise efficient landscape-oriented management strategies for invasive species and can also generate insights for conservation purposes.

Hypovitaminosis D and hypervitaminosis D are well recognised disorders in dogs. Hypovitaminosis D can occur following consumption of a diet inadequately supplemented with vitamin D or as a sequelae of severe intestinal disease. Hypervitaminosis D may occur as a result of consuming proprietary dog foods over-supplemented with vitamin D or through ingestion of vitamin D containing medicinal products or rodenticides. Consequently, there is a clear need to establish a methodology that can accurately quantify vitamin D metabolites across a broad dynamic range in dogs. The existence of C3-epimers of vitamin D metabolites has yet to be elucidated in dogs, yet are known to interfere with the analysis of vitamin D and have unknown biological activity in other species. Here, we describe the development and validation of a sensitive, specific and robust analytical liquid chromatography tandem mass spectrometry (LC–MS/MS) assay capable of separating and accurately measuring 25-hydroxyvitamin-D2/3 (25(OH)D2/3) and 3-epi-25-hydroxyvitamin-D2/3 (3-epi-25(OH)D2/3). We describe a simplified workflow utilising supported liquid extraction (SLE) without derivatization that provides good linearity (mean r > 0.996) and accuracy across a broad dynamic range of 4–500 nmol/L for D3 metabolites and 7.8–500 nmol/L for D2 metabolites. Upon application of this assay to 117 canine serum samples, 25(OH)D3 was detectable in all samples with a median concentration of 82.1 nmol/L (inter-quartile range (IQR) 59.7–101.8 nmol/L). 3-epi-25(OH)D3 could be detected in 87.2 % of the study population, with a median concentration of 5.2 nmol/L (2.4–8.1 nmol/L). However, 3-epi-25(OH)D3 was quantified below the LLOQ in 40.2 % of these samples. 3-epi-25(OH)D3 contributed on average 6.3 % to 25(OH)D3 status (contribution ranges from 0 to 23.8%) and a positive correlation was detected between 25(OH)D3 and 3-epi-25(OH)D3 concentrations. Free 25(OH)D was also measured using an immunoassay with a median concentration of 15.2 pmol/L (12.5–23.2 pmol/L), and this metabolite was also positively correlated to both 3-epi-25(OH)D3 and 25(OH)D3 concentrations. D2 metabolites were not detected in canine serum as expected. Vitamin D metabolite concentrations were variable between individuals, and research into the causes of this variation should include factors such as breed, age, sex and neuter status to determine the impact of genetic and hormonal factors. Given the clinical importance of vitamin D in dogs, and the immense potential for utilising this species as a model for human disease, further elucidation of the vitamin D pathway in this species would provide immense clinical and research benefit.

The choice of day roosts by microbats influences energetics, social interactions and breeding success. In Australia, the large-footed myotis (Myotis macropus) is dependent on waterways for foraging. However, the extent to which the species relies upon, and selects roosts within, riparian habitat is unknown. I studied the roosting behaviour of this species around a water reservoir near Melbourne, Victoria, during the summers from October 2002 to April 2005. I radio-tracked 31 bats to 17 tree roosts; colony size averaged 6.0 ± 1.6 bats and individuals used 1.3 ± 0.1 roosts during the tracking period (average 6.2 ± 0.6 days). Two roosts were also located in crevices in an old aqueduct tunnel, housing colonies of 10.8 ± 0.6 bats. Colonies emerged earlier from tunnel crevices (25.0 ± 3.6 min after sunset, range 6–53 min), compared with conspecifics in tree roosts (45.6 ± 2.1 min after sunset, range 10–83 min). Roost trees and cavities differed from available habitat trees and cavities in terms of smaller entrance areas to used (182.2 ± 49.3 cm2) versus unused (328.0 ± 61.8 cm2) cavities. The primary force driving roost selection by M. macropus appears to be proximity of suitable waterways for foraging. Retention and maintenance of extensive riparian habitat, as well as the preservation of other structures used for roosting, are the most important conservation strategies for management of the day-roosting habitat of M. macropus.

Background. Most chronic pain patients are treated in primary care and their management is often challenging. Secondary care- or private sector-based Pain Management Programmes (PMPs) offering intensive multidisciplinary approaches have been found to improve participants' physical performance and psychological well-being.

The Judas technique is a method used for landscape control of feral donkeys (Equus asinus) in northern Australia. Central to the success of any Judas program is the safe, efficient, and humane attachment of the telemetry device. For feral donkeys, this involves the use of field immobilization. We examine the replacement of the current chemical capture agent, succinylcholine, with contemporary immobilization agents to achieve positive animal welfare outcomes. A combination of medetomidine and ketamine delivered by remote injection from a helicopter was used to capture 14 free-ranging feral donkeys for the fitting of telemetry collars in Western Australia in November 2010. Dose rates of 0.14 mg/kg medetomidine and 4.1 mg/kg ketamine were appropriate to immobilize animals in 9 min (± SD = 3). Mean recovery time (total time in recumbency) was 21 min (± 14). All animals recovered uneventfully after being administered atipamezole, a specific antagonist of medetomidine, intramuscularly at 0.35 mg/kg. Physiologic parameters were recorded during recumbency, with environment-related hyperthermia being the only abnormality recognized. No significant complications were encountered, and this drug combination represents an efficient approach to capturing wild donkeys. This new method allows a rapid, safe, cost-effective approach to the immobilization of feral donkeys for use as Judas animals. This drug combination will replace the relatively inhumane succinylcholine for the field immobilization of feral donkeys.

Pain Management Programmes (PMPs) are a multi-disciplinary approach to the management of chronic low back pain (CLBP). Notwithstanding evidence of effectiveness, successful take-up of programmes requires acceptability to patients. We used a discrete choice experiment to investigate patient preferences for alternative PMPs for managing CLBP in primary care. Specifically, we estimated the probability of uptake of alternative configurations of PMPs. Potential attributes and associated levels influencing take-up were identified through a systematic literature review, survey of current PMPs, expert consultation, and focus groups. Five attributes were included: content; provider; schedule; group size; and travel time to clinic. Four hundred and fourteen questionnaires were mailed to patients attending clinics and 124 questionnaires were returned suitable for analysis. Method of delivery influenced probability of take-up, with small group sizes and low intensity programmes over a prolonged period increasing the probabilities. Travel time was also important. However, providers and contents of PMPs were not main drivers of preferences, though those with more severe pain did prefer PMPs provided by more specialists. Probability of take-up increases when PMPs better reflect patient preferences. Given preferences, resource constraints, and evidence on clinical outcomes of alternative configurations it is suggested more resource-intensive PMPs be reserved for those with the most severe and disabling pain and less intensive programmes delivered over a longer time period in smaller groups for those with less severe pain. These findings can inform future randomised trials to evaluate acceptable PMPs in primary care.

Juvenile segmental progeroid syndromes are rare, heterogeneous disorders characterized by signs of premature aging affecting more than one tissue or organ starting in childhood. Hutchinson–Gilford progeria syndrome (HGPS), caused by a recurrent de novo synonymous LMNA mutation resulting in aberrant splicing and generation of a mutant product called progerin, is a prototypical example of such disorders. Here, we performed a joint collaborative study using massively parallel sequencing and targeted Sanger sequencing, aimed at delineating the underlying genetic cause of 14 previously undiagnosed, clinically heterogeneous, non-LMNA-associated juvenile progeroid patients. The molecular diagnosis was achieved in 11 of 14 cases (~ 79%). Furthermore, we firmly establish biallelic mutations in POLR3A as the genetic cause of a recognizable, neonatal, Wiedemann–Rautenstrauch-like progeroid syndrome. Thus, we suggest that POLR3A mutations are causal for a portion of under-diagnosed early-onset segmental progeroid syndromes. We additionally expand the clinical spectrum associated with PYCR1 mutations by showing that they can somewhat resemble HGPS in the first year of life. Moreover, our results lead to clinical reclassification in one single case. Our data emphasize the complex genetic and clinical heterogeneity underlying progeroid disorders.

Purpose.: To assess the effects of body stature and years of education, in addition to age and sex, on six oculometric traits and to estimate the heritabilities of these quantitative traits in two Croatian cross-population studies. Methods.: Adult subjects living on the two Croatian islands of Vis and Korčula were recruited for a large epidemiologic and genetic study that included eye biometry, keratometry, and autorefraction. Effects and heritabilities were estimated by using general linear mixed models for axial length (AL), anterior chamber depth (ACD), corneal curvature (CC), corneal thickness (CT), lens thickness (LT), and spherical equivalent refraction (SER). Both cohorts were genotyped with dense SNP arrays, allowing the use of kinship coefficients derived from genotypic data (realized kinship) rather than from pedigree information (expected kinship). Results.: Across cohorts, body mass index (BMI) did not consistently influence any of the ocular traits adjusted for age and/or sex, whereas height and years in education (YrEd) did, explaining up to an additional 5% of the variance (in CC). CT was the trait least influenced by covariates. Estimated heritabilities in Vis and Korčula, respectively, were 84% and 52% for CC, 75% and 71% for CT, 37% and 32% for LT, 59% and 45% for ACD, 37% and 74% for AL, and 0% and 17% for SER. Conclusions.: While heritabilities of CT and CC seemed uniformly high across studies of Caucasian datasets, estimates for SER varied widely and were at the lower end of the spectrum of published observations in our study.

Specialist species face higher extinction risks as a result of smaller, isolated populations with reduced gene flow. The large-footed myotis (Myotis macropus) is the only microbat in Australia specialised for foraging directly over water surfaces. Such highly specialised feeding ecology restricts the distribution of M. macropus to coastal regions and inland waterways. Using five novel and two existing nuclear microsatellite markers, we investigated genetic diversity within and among five M. macropus populations in Victoria. Significant genetic differentiation was detected between all populations. FST values between populations ranged from 0.02 to 0.24. We suggest that the movement of M. macropus throughout the landscape is constrained by the availability of permanent waterways and associated riparian habitats. These findings represent important considerations for the conservation of this specialist species and the management of riparian vegetation, particularly on private land.

Measures that reflect patients' assessment of their health are of increasing importance as outcome measures in randomised controlled trials. The methodological approach used in the pre-validation development of new instruments (item generation, item reduction and question formatting) should be robust and transparent. The totality of the content of existing PRO instruments for a specific condition provides a valuable resource (pool of items) that can be utilised to develop new instruments. Such 'top down' approaches are common, but the explicit pre-validation methods are often poorly reported. This paper presents a systematic and generalisable 5-step pre-validation PRO instrument methodology.The method is illustrated using the example of the Aberdeen Glaucoma Questionnaire (AGQ). The five steps are: 1) Generation of a pool of items; 2) Item de-duplication (three phases); 3) Item reduction (two phases); 4) Assessment of the remaining items' content coverage against a pre-existing theoretical framework appropriate to the objectives of the instrument and the target population (e.g. ICF); and 5) qualitative exploration of the target populations' views of the new instrument and the items it contains.The AGQ 'item pool' contained 725 items. Three de-duplication phases resulted in reduction of 91, 225 and 48 items respectively. The item reduction phases discarded 70 items and 208 items respectively. The draft AGQ contained 83 items with good content coverage. The qualitative exploration ('think aloud' study) resulted in removal of a further 15 items and refinement to the wording of others. The resultant draft AGQ contained 68 items.This study presents a novel methodology for developing a PRO instrument, based on three sources: literature reporting what is important to patient; theoretically coherent framework; and patients' experience of completing the instrument. By systematically accounting for all items dropped after the item generation phase, our method ensures that the AGQ is developed in a transparent, replicable manner and is fit for validation. We recommend this method to enhance the likelihood that new PRO instruments will be appropriate to the research context in which they are used, acceptable to research participants and likely to generate valid data.

To develop patient-reported outcome instruments, statistical techniques (e.g., principal components analysis; PCA) are used to examine correlations among items and identify interrelated item subsets (empirical factors). However, interpretation and labelling of empirical factors is a subjective process, lacking precision or conceptual basis. We report a novel and reproducible method for mapping between theoretical and empirical factor structures. We illustrate the method using the pilot Aberdeen Glaucoma Questionnaire (AGQ), a new measure of glaucoma-related disability developed using the International Classification of Functioning and Disability (ICF) as a theoretical framework and tested in a sample representing the spectrum of glaucoma severity.We used the ICF to code AGQ item content before mailing the AGQ to a UK sample (N = 1349) selected to represent people with a risk factor for glaucoma and people with glaucoma across a range of severity. Reflecting uncertainty in the theoretical framework (items with multiple ICF codes), an exploratory PCA was conducted. The theoretical structure informed our interpretation of the empirical structure and guided the selection of theoretically-derived factor labels. We also explored the discrimination of the AGQ across glaucoma severity groups.656 (49%) completed questionnaires were returned. The data yielded a 7-factor solution with a simple structure (using cut-off point of a loading of 0.5) that together accounted for 63% of variance in the scores. The mapping process resulted in allocation of the following theoretically-derived factor labels: 1) Seeing Functions: Participation; 2) Moving Around & Communication; 3) Emotional Functions; 4) Walking Around Obstacles; 5) Light; 6) Seeing Functions: Domestic & Social Life; 7) Mobility. Using the seven factor scores as independent variables in a discriminant function analysis, the AGQ scores resulted in correct glaucoma severity grading of 32.5% of participants (p < 0.001).This paper addresses a methodological gap in the application of classical test theory (CTT) techniques, such as PCA, in instrument development. Labels for empirically-derived factors are often selected intuitively whereas they can inform existing bodies of knowledge if selected on the basis of theoretical construct labels, which are more explicitly defined and which relate to each other in ways that are evidence based.

Transient receptor potential vanilloid family member 4 (TRPV4) transcript and protein expression increased in the urinary bladder and lumbosacral dorsal root ganglia of transgenic mice with chronic urothelial overexpression of nerve growth factor (NGF-OE). We evaluated the functional role of TRPV4 in bladder function with open-outlet cystometry, void spot assays, and natural voiding (Urovoid) assays with the TRPV4 antagonist HC-067047 (1 μM) or vehicle in NGF-OE and littermate wild-type (WT) mice. Blockade of TRPV4 at the level of the urinary bladder significantly (P ≤ 0.01) increased the intercontraction interval (2.2-fold) and void volume (2.6-fold) and decreased nonvoiding contractions (3.0-fold) in NGF-OE mice, with lesser effects (1.3-fold increase in the intercontraction interval and 1.3-fold increase in the void volume) in WT mice. Similar effects of TRPV4 blockade on bladder function in NGF-OE mice were demonstrated with natural voiding assays. Intravesical administration of HC-067047 (1 µM) significantly (P ≤ 0.01) reduced pelvic sensitivity in NGF-OE mice but was without effect in littermate WT mice. Blockade of urinary bladder TRPV4 or intravesical infusion of brefeldin A significantly (P ≤ 0.01) reduced (2-fold) luminal ATP release from the urinary bladder in NGF-OE and littermate WT mice. The results of the present study suggest that TRPV4 contributes to luminal ATP release from the urinary bladder and increased voiding frequency and pelvic sensitivity in NGF-OE mice.

Traces of microboring microbial endoliths (euendoliths) conform closely to the outlines of their makers as they are made. This habit evolved among prokaryotic cyanobacteria and eukaryotic rhodophytes, chlorophytes, and fungi. Among eukaryotic microborers, the endolithic mode of life is often limited to a phase in the course of their development, which alternates with an epilithic leafy phase. These endolithic phases are sometimes independently described as separate genera and species. This was the case with Conchocelis rosea Batters, 1891, later identified as the endolithic phase of Porphyra and Bangia that helped in identification of fossil bangialean rhodophytes of Silurian age: Palaeoconchocelis starmachii Campbell, Kazmierczak et Golubic, 1979. However, the traces of Conchocelis phases, although conspicuous and used to characterize the resident body fossil, were never formally described. We present here the formal ichnotaxonomic description of the complex and variable Conchocelis trace as Conchocelichnus seilacheri igen. et isp. nov., based primarily on Oligocene (Tertiary) borings in molluscan shells of Mainz Basin, Germany, while consulting similar fossil borings of the Silurian and Jurassic findings and compareing them with modern occurrences in nature and culture. With the new trace name Conchocelichnus seilacheri igen. et isp. nov., we honor the work of the renowned paleontologist and ichnologist, Prof. Dr. Dolf Seilacher.

Abstract With ongoing introductions into Australia since the 1700s, the European rabbit ( Oryctolagus cuniculus ) has become one of the most widely distributed and abundant vertebrate pests, adversely impacting Australia's biodiversity and agroeconomy. To understand the population and range dynamics of the species and its impacts better, occurrence and abundance data have been collected by researchers and citizens from sites covering a broad spectrum of climatic and environmental conditions in Australia. The lack of a common and accessible repository for these data has, however, limited their use in determining important spatiotemporal drivers of the structure and dynamics of the geographical range of rabbits in Australia. To meet this need, we created the Australian National Rabbit Database, which combines more than 50 yr of historical and contemporary survey data collected from throughout the range of the species in Australia. The survey data, obtained from a suite of complementary monitoring methods, were combined with high‐resolution weather, climate, and environmental information, and an assessment of data quality. The database provides records of rabbit occurrence (689,265 records) and abundance (51,241 records, &gt;120 distinct sites) suitable for identifying the spatiotemporal drivers of the rabbit's distribution and for determining spatial patterns of variation in its key life‐history traits, including maximum rates of population growth. Because all data are georeferenced and date stamped, they can be coupled with information from other databases and spatial layers to explore the potential effects of rabbit occurrence and abundance on Australia's native wildlife and agricultural production. The Australian National Rabbit Database is an important tool for understanding and managing the European rabbit in its invasive range and its effects on native biodiversity and agricultural production. It also provides a valuable resource for addressing questions related to the biology, success, and impacts of invasive species more generally. No copyright or proprietary restrictions are associated with the use of this data set other than citation of this Data Paper.

Purpose: Family history is the most prominent risk factor, besides advanced age, for the incidence of breast cancer among women. This study investigates differences in the experiences of women in the detection, diagnosis, and treatment of early‐stage disease. The purpose of this research is to obtain a more comprehensive understanding of the impact of family history on the overall illness experience. Description of study: Self‐report retrospective data obtained from in‐depth interviews with a convenience sample of 179 women who had recently received a diagnosis of nonrecurrent stage 0 to IIIA breast cancer are used to compare the experiences of women with and without a family history of breast cancer (FHOBC). The authors examine differences in screening behavior, method of detection, diagnostic processes, treatment decision making, and therapy receipt, and they report the results of bivariate analyses. Results: The results suggest that women with FHOBC have a different disease experience than those without an affected relative. Women with FHOBC were more likely than their counterparts to comply with screening guidelines, to seek more timely care, to consult with specialists, to be influenced by the experiences of others, to feel comfortable with treatment decisions, and to receive adjuvant therapy. Clinical implications: Healthcare providers should be aware that compliance with mammography and therapy guidelines may vary with FHOBC. Because the better health‐related behavior reported by women with affected relatives suggests that they may have higher perceived risk, physicians should be sensitive to potentially elevated levels of anxiety, provide accurate information about relative risk, put patient concerns in the proper perspective, and include family members in treatment discussions. Alternatively, women without an FHOBC appear to have less favorable screening, detection, diagnosis, and treatment decision‐making behavior. Because family doctors play an important role in the care of these patients, they may need to provide special education and counseling regarding the importance of adherence to screening guidelines, recognition of relevant symptoms, initiation of timely examinations, consultation with cancer specialists, and compliance with treatment recommendations.

We have used targeted genomic sequencing of high-complexity DNA pools based on long-range PCR and deep DNA sequencing by the SOLiD technology. The method was used for sequencing of 286 kb from four chromosomal regions with quantitative trait loci (QTL) influencing blood plasma lipid and uric acid levels in DNA pools of 500 individuals from each of five European populations. The method shows very good precision in estimating allele frequencies as compared with individual genotyping of SNPs (r(2) = 0.95, P < 10(-16)). Validation shows that the method is able to identify novel SNPs and estimate their frequency in high-complexity DNA pools. In our five populations, 17% of all SNPs and 61% of structural variants are not available in the public databases. A large fraction of the novel variants show a limited geographic distribution, with 62% of the novel SNPs and 59% of novel structural variants being detected in only one of the populations. The large number of population-specific novel SNPs underscores the need for comprehensive sequencing of local populations in order to identify the causal variants of human traits.

Purpose of the study:The Alpert Medical School of Brown University began to integrate geriatrics content into all preclerkship courses and key clerkship cases as part of a major medical school curriculum redesign in 2006. This study evaluates students’ responses to geriatrics integration within the curriculum using journals kept by volunteer preclerkship and clerkship students between 2007 and 2010. The journals were used to assess the quality of curricular integration of geriatrics didactic and clinical content, to gather information for shaping the evolving curriculum, and to elicit students’ responses about their professional development and caring for older adults.

Glaucoma is a leading cause of blindness. Early detection is advocated but there is insufficient evidence from randomized controlled trials (RCTs) to inform health policy on population screening. Primarily, there is no agreed screening intervention. For a screening programme, agreement is required on the screening tests to be used, either individually or in combination, the person to deliver the test and the location where testing should take place. This study aimed to use ophthalmologists (who were experienced glaucoma subspecialists), optometrists, ophthalmic nurses and patients to develop a reduced set of potential screening tests and testing arrangements that could then be explored in depth in a further study of their feasibility for evaluation in a glaucoma screening RCT. A two-round Delphi survey involving 38 participants was conducted. Materials were developed from a prior evidence synthesis. For round one, after some initial priming questions in four domains, specialists were asked to nominate three screening interventions, the intervention being a combination of the four domains; target population, (age and higher risk groups), site, screening test and test operator (provider). More than 250 screening interventions were identified. For round two, responses were condensed into 72 interventions and each was rated by participants on a 0-10 scale in terms of feasibility. Using a cut-off of a median rating of feasibility of ≥5.5 as evidence of agreement of intervention feasibility, six interventions were identified from round 2. These were initiating screening at age 50, with a combination of two or three screening tests (varying combinations of tonometry/measures of visual function/optic nerve damage) organized in a community setting with an ophthalmic trained technical assistant delivering the tests. An alternative intervention was a ‘glaucoma risk score’ ascertained by questionnaire. The advisory panel recommended that further exploration of the feasibility of screening higher risk populations and detailed specification of the screening tests was required. With systematic use of expert opinions, a shortlist of potential screening interventions was identified. Views of users, service providers and cost-effectiveness modeling are now required to identify a feasible intervention to evaluate in a future glaucoma screening trial.

Purpose Elder abuse results in high rates of morbidity and mortality. It has longstanding physical and psychological effects and is difficult to detect. Due to fear or embarrassment, victims may make attempts to hide it rather than to disclose and professionals are often reluctant to report it as they may worry about worsening a situation. If detected early enough, serious harm can be prevented and lives saved. Screening and screening tools can assist health and social care practitioners to detect abuse. This review of screening tools was undertaken as part of an MSc in clinical research, funded by the National Institute for Health Research; the purpose of this paper is to report on the review and its findings. Design/methodology/approach This was a systematic review with eligibility inclusion and exclusion criteria decided in advance. Keywords and their synonyms were combined and then used to search health and social care databases. Data items were collected from the included studies. The preferred reporting item for systematic reviews and meta-analysis was followed for the reporting of the results. A narrative synthesis approach was applied to the analysis. Findings A total of 34 full text studies were downloaded, read and analysed. In all, 11 met the inclusion criteria and were included in the final analysis. Of these, three studies reported sensitivity and specificity, with the remainder reporting validity and reliability testing. In total, 12 tools of varying length and quality were found. The length and characteristics of tools affects the efficacy of their use. The clinical environment will determine choice of screening tool to be used. Screening tools should be used within an overall system of detection and management of abuse. Research limitations/implications The synthesis of results was challenging due to the lack of homogeneity between the included studies. The variations in tool characteristics and qualities added to this challenge. A further limitation was the lack of a gold standard tool in elder abuse. Originality/value This systematic review highlights a lack of robust evidence in the development and validation of screening tools to detect elder abuse. Though there is an increasing awareness and knowledge about elder abuse, its detection remains problematic and the lack of research in this area is worth emphasising. Specific tools, centred on the clinical setting in which they are used, are recommended.

BackgroundHutchinson-Gilford progeria syndrome (HGPS) is an ultra-rare, fatal, segmental premature aging syndrome caused by the aberrant lamin A protein, progerin. The protein farnesyltransferase inhibitor, lonafarnib, ameliorates some aspects of cardiovascular and bone disease.MethodsWe performed a prospective longitudinal survey of plasma proteins in 24 children with HGPS (an estimated 10% of the world's population at the time) at baseline and on lonafarnib therapy, compared with age- and gender-matched controls using a multi-analyte, microsphere-based immunofluorescent assay.ResultsThe mean levels for 23/66 (34.8%) proteins were significantly lower and 7/66 (10.6%) were significantly higher in HGPS samples compared with those in controls (P≤0.05). Six proteins whose concentrations were initially lower normalized with lonafarnib therapy: interleukins 1α, 7, and 13, beta-2 microglobulin, C-reactive protein, and myoglobin. Alpha-2 macroglobulin, a protease inhibitor associated with stroke, was elevated at baseline and subsequently normalized with lonafarnib therapy.ConclusionThis is the first study to employ a multi-analyte array platform in HGPS. Novel potential biomarkers identified in this study should be further validated by correlations with clinical disease status, especially proteins associated with cardiovascular disease and those that normalized with lonafarnib therapy.

The increasing popularity of citizen science in ecological research has created opportunities for data collection from large teams of observers that are widely dispersed. We established a citizen science program to complement the release of a new variant of the rabbit biological control agent, rabbit haemorrhagic disease virus (RHDV), known colloquially as K5, across Australia. We evaluated the impact of K5 on the national rabbit population and compared citizen science and professionally-collected spotlight count data. Of the citizen science sites (n = 219), 93% indicated a decrease in rabbit abundance following the release of K5. The overall finite monthly growth rate in rabbit abundance was estimated as 0.66 (95%CI, 0.26, 1.03), averaging a monthly reduction of 34% at the citizen science sites one month after the release. No such declines were observed at the professionally monitored sites (n = 22). The citizen science data submissions may have been unconsciously biased or the number of professional sites may have been insufficient to detect a change. Citizen science participation also declined by 56% over the post-release period. Future programs should ensure the use of blinded trials to check for unconscious bias and consider how incentives and/or the good will of the participants can be maintained throughout the program.

The European rabbit (Oryctolagus cuniculus) is a notorious economic and environmental pest species in its invasive range. To better understand the population and range dynamics of this species, 41 yr of abundance data have been collected from 116 unique sites across a broad range of climatic and environmental conditions in Australia. We analyzed this time series of abundance data to determine whether interannual variation in climatic conditions can be used to map historic, contemporary, and potential future fluctuations in rabbit abundance from regional to continental scales. We constructed a hierarchical Bayesian regression model of relative abundance that corrected for observation error and seasonal biases. The corrected abundances were regressed against environmental and disease variables in order to project high spatiotemporal resolution, continent-wide rabbit abundances. We show that rabbit abundance in Australia is highly variable in space and time, being driven primarily by internnual variation in temperature and precipitation in concert with the prevalence of a non-pathogenic virus. Moreover, we show that internnual variation in local spatial abundances can be mapped effectively at a continental scale using highly resolved spatiotemporal predictors, allowing "hot spots" of persistently high rabbit abundance to be identified. Importantly, cross-validated model performance was fair to excellent within and across distinct climate zones. Long-term monitoring data for invasive species can be used to map fine-scale spatiotemporal fluctuations in abundance patterns when accurately accounting for inherent sampling biases. Our analysis provides ecologists and pest managers with a clearer understanding of the determinants of rabbit abundance in Australia, offering an important new approach for predicting spatial abundance patterns of invasive species at the near-term temporal scales that are directly relevant to resource management.

Abstract NOTE: The first page of text has been automatically extracted and included below in lieu of an abstract Session Number 3530 Teaching and Assessing Engineering Design: A Review of the Research Susan Campbell Carol L. Colbeck The Pennsylvania State University According to the National Academy of Sciences (1995), undergraduate engineering education in the United States currently focuses on the study of engineering science at the expense of design. In a brief history of engineering design education, Eder (1991) explains that in the 1950s the engineering curricula shifted from a focus on teaching students about technology used in industry to an emphasis on engineering science including math, physics, and chemistry. He contends that faculty began to become more research-oriented and institutions of higher education placed less value on industry experience among faculty at roughly the same time. Eder argues that these factors contributed to the recent lack of emphasis on design skills and abilities in engineering curricula. He attributes renewed attention to design in curricula to a 1985 National Science Foundation initiative that resulted in funding to improve undergraduate engineering design education. The Accreditation Board for Engineering Technology (ABET) and many industry representatives join the National Academy of Sciences in calling for a renewed emphasis on the development of design skills among undergraduate engineering students. ABET stipulates that graduates of accredited engineering programs demonstrate skills and competencies such as an “ability to design a system, component, or process; an ability to function on multidisciplinary teams; an ability to identify, formulate, and solve engineering problems;...[and] an ability to communicate effectively” (ASEE Prism, 1997, p. 41). Likewise, industry representatives emphasize that undergraduate engineering students should develop an understanding of the design process as well as the ability to work in teams, communicate effectively, think critically, and solve problems (Rhoads, et al., 1995; Coleman, 1996). This paper begins with a definition of engineering design followed by a summary of the research on teaching design and assessing student design competence and concludes with implications for future research to improve engineering education. Definition of engineering design Engineering design is complex problem solving (Lewis & Samuel, 1989) that involves generating and evaluating specifications to achieve objectives and satisfy constraints (Dym, 1994). This definition implies that: (1) engineering design is a cognitive process that can be modeled and understood; (2) representations exist for both the form and function of the processes involved in design; (3) the objectives of the design problem and any constraints can be determined from this representation; (4) design alternatives can be generated using problem- solving techniques; (5) fabrication specifications can be determined from the designs; and (6) designs can be assessed and evaluated at various points in the design process (Dym, 1994).

Small cell lung cancer (SCLC) is a highly invasive and metastatic tumor, and the decreased expression of alpha3beta1 integrin may contribute to its virulence. Alpha3beta1 is a critical integrin for pulmonary development and epithelial integrity, and its reduced expression has been linked to the increased malignancy and invasion of other cancers. The amplification of the c-myc oncogene is seen frequently in relapsed SCLC tumors and is associated with a worsened prognosis. In the present study using a model of SCLC tumor progression, overexpression of c-myc in a classic SCLC cell line, NCI H209, enhanced in vitro features of tumorigenesis, altered the relationships between cell and environment, and markedly down-regulated the expression of the alpha3 integrin subunit at both the transcript and protein levels. This inverse relationship between the expression of the alpha3 integrin subunit and c-myc is mimicked by other c-myc-overexpressing SCLC cell lines. Restoring alpha3 expression in the myc-transfected 209 cells reversed the effects of c-myc: alpha3 transfection increased cell:cell adhesion and reduced soft agar cloning without affecting the in vitro doubling time. The diminished soft agar cloning produced by alpha3 transfection was reversed by an antibody that specifically engages alpha3beta1 integrins, P1B5. These results suggest first, that alpha3beta1 integrin mediates homotypic adhesion of SCLC cells, and second, that unengaged alpha3beta1 integrin suppresses the growth of disaggregated SCLC cells. Thus, the down-regulation of the alpha3 integrin subunit may contribute to the enhanced tumorigenicity of c-myc-overexpressing SCLCs by allowing the growth of tumor cells that have reduced contact with ligand-expressing substratum or cells, a condition that occurs during the growth of the primary tumor, tumor invasion, and metastasis.

The day roosting behaviour of the little forest bat (Vespadelus vulturnus), Australia’s smallest bat, was investigated in the context of the planned removal of dead timber within managed woodlands on Phillip Island, Victoria. Between August 1999 and March 2000, 14 female little forest bats were fitted with VHF microtransmitters and tracked to a total of 16 roost trees. All roosts were located in dead timber, 11 in severely decayed remains of eucalypt trees, and five in dead sections of live trees. Roost trees were compared with randomly chosen trees from within the available habitat, for a range of tree characteristics. Female little forest bats selected roosts in trees with dead timber offering many hollows and reduced canopy cover. Furthermore, roost trees were located in areas (0.1-ha plots) with higher densities of these types of trees than in the available habitat. However, there was no difference in the height or diameter of roost trees or roost plots compared with available habitat. Emergence time from roosts was strongly associated with civil twilight (when the centre of the sun is 6° below an ideal horizon), and the number of bats exiting a single roost tree ranged from 1 to 120 (median = 20). Dead trees provide critical roosting habitat and we recommend retention of dead standing trees for conserving little forest bat roosts in managed woodlands.

ABSTRACT: Context: Since the passage of the Balanced Budget Act of 1997, rural hospitals have struggled with the need to strategically adapt to an abundance of changing reimbursement and regulatory programs, as well as to respond to the needs of an increasingly frail elder population in need of postacute and long‐term care (LTC). Purpose: This article has 2 goals: (1) to provide a summary of the many legislative acts and provisions influencing rural hospital LTC strategies during the 1997‐2003 period and (2) to track changes in the LTC strategies of a national sample of rural hospitals through this 7‐year period. Methods: A 3‐wave panel of rural hospital discharge planners in 540 nonfederal community‐general hospitals were interviewed in 1997, 2000, and 2003. Questions focused on hospital structure, discharge planning process, and reports of internal and external organizational arrangements for providing LTC services to hospitalized patients, and changes in LTC strategy since the previous interview. Descriptive statistics are presented on LTC strategies in place in 1997 and dropped or added in 2000 and 2003. Findings and Conclusions: The general shape of the regulatory environment confronting rural hospitals and their LTC strategies during the recent past can be described as complicated, rapidly changing, and at times contradictory in intended effects. There has been a large volume of strategy change during this 7‐year period, without the emergence of any identifiable pattern or LTC strategy profile, other than swing‐bed participation combined with home health agency ownership.

Reflective writing techniques such as journaling help provide insights into the process by which medical students are mentored and develop into practicing physicians. The authors sought to analyze medical students' journals regarding their mentored experiences within a new geriatrics curriculum at a U.S. medical school. Thirty preclinical and clinical medical student journalers participated in this project. The authors employed qualitative analytic techniques using an interdisciplinary team process. Three major themes emerged: (a) exposure to clinical mentors challenged medical students' preconceptions regarding older adults and geriatric medicine; (b) students learned new medical knowledge and techniques from observing their mentors; and (c) students provided positive and negative assessments of their mentors. Reflective journaling provides important insights into the process by which medical students draw upon mentored clinical experiences during their training. Such mentorship may be particularly relevant to promoting their interest in geriatrics.

Psychological stress is associated with urinary bladder dysfunction (e.g., increased voiding frequency, urgency and pelvic pain); however, the mechanisms underlying the effects of stress on urinary bladder function are unknown. Transient receptor potential (TRP) channels (vanilloid family) may be potential targets for intervention due to their distribution in the LUT and role in pain. Here, we examine a model of repeated variate stress (RVS) of 2 week (wk) or 4 wk duration in female mice and its effects on bladder function, anxiety-like behavior, and TRPV transcript expression in urinary bladder and lumbosacral spinal cord and associated dorsal root ganglia (DRG). Using continuous infusion, open-outlet cystometry in conscious mice, RVS significantly (p ≤ 0.05) decreased infused volume and intermicturition interval. Bladder pressures (threshold, average, minimum, and maximum pressures) were unchanged with RVS. Quantitative PCR demonstrated significant (p ≤ 0.05) changes in TrpV1 and TrpV4 mRNA expression between control and RVS cohorts in the urothelium, lumbosacral spinal cord, and DRG. Future directions will examine the contribution of TRP channels on bladder function, somatic sensation and anxiety-like behavior following RVS.

Six middle school students diagnosed with attention deficit/hyperactivity disorder were selected for sensorimotor rhythm (SMR) training with EEG biofeedback. The subjects were evaluated following a 72-hour drug-free period with the WISC-III Digit Span subtest and the Test of Variables of Attention (TOVA). Five of the subjects received 20 sessions of EEG biofeedback and one of the subjects received nine sessions of EEG biofeedback. The subjects were evaluated again following a 72-hour drug-free period. Five of the six subjects improved on their combined Digit Span, TOVA Inattention, and TOVA Impulsivity scores. These results supported previous findings that EEG biofeedback can be effective in the treatment of attention deficit/hyperactivity disorder. More importantly, this study demonstrated that EEG biofeedback could be used in an actual school setting. Recommendations for implementing an EEG biofeedback program in the schools were provided.

Brown Medical School developed a comprehensive curriculum in which enriched aging content increased from 22 to 80 hours in preclerkship courses and was also added for clerkships, residencies, and nongeriatrician physicians. Innovative evaluation strategies are also described. Highlights include “treasure hunts” in the anatomy laboratory, a Scholarly Concentration in Aging, Schwartz Communication Sessions, a Website of aging-related materials, and a monthly column in the state medical journal. Evaluation includes “tracking” to compute the “dose” of aging content, and “journaling” and focus groups to evaluate students' responses. Integrating geriatrics across a broad range of courses and clinical experiences is feasible.

The thermal environment of day roosts is considered one of the most influential factors affecting the survival, growth and reproduction of microbats. The use of torpor is a common energy saving strategy employed by microbats in temperate regions. The efficiency of entry into, and arousal from, torpor is governed by roost microclimate, primarily roost temperature. The large-footed myotis Myotis macropus roosts in both tree cavities and a man-made tunnel at Yan Yean reservoir in Victoria, Australia. We investigated the thermal properties of both roost types in comparison to available tree cavities and ambient temperature over four time periods from October 2003 to May 2005. Tree cavities and tunnel roosts remained significantly warmer at night, cooler during the day, and were more stable than ambient temperatures. In addition, roost tree cavities were significantly cooler between 10:00–13:00 h compared to available tree cavities, and there was a trend for roost tree cavities to be slightly warmer at night and slower to reach maximum temperature relative to available tree cavities during the breeding season (October–January). In contrast, there was little difference in roost and available tree cavity temperatures outside of the breeding season (April–May). Temperatures inside tunnel roosts were more stable and were significantly cooler during the afternoon compared to roost tree cavities during both the breeding and non-breeding seasons. Myotis macropus may actively trade-off the enhanced thermoregulatory benefits of warm roosts for reduced predation risk associated with the tunnel roosting environment.

A complex microboring trace of fungal affinity is described in shells as a new ichnotaxon Saccomorpha stereodiktyon isp. nov. and compared with the earlier established ichnotaxon Saccomorpha terminalis Radtke, 1991. The new trace is characterized by a three-dimensional network of tunnels composed of a bifurcate horizontal (parallel to substrate surface) network with an upright (perpendicular to surface) system of tunnels and by the formation of cylindrical to multilobate terminal sporangial swellings. The new trace shares with Saccomorpha terminalis Radtke, 1991 the terminal position of sporangial swellings but differs from this ichnotaxon by its complexity in spatial arrangement, segmented construction, and ramification of tunnels. The horizontal parts of the network in the new taxon adhere to the substrate surface and regularly produce thinner tunnels that explore the interior of the substrate, allowing the producer to participate in digestion of organic lamellae incorporated in the shell. Microborings similar to the new trace fossil have been observed in modern bivalve shells of the Atlantic Ocean, North Sea, Adriatic Sea and Red Sea at depths ranging from the intertidal down to 1,550 m. The fossil record of the trace reaches back to the Jurassic and the type material stems from a Lower Oligocene oyster shell. The study shows that complex microboring traces reflect both behaviour and developmental strategy of their makers.

The overexpression of c-myc frequently accompanies the relapse of small cell lung cancer (SCLC) cells and contributes to the poor prognosis of this tumor. In this study, we confirm that transfected c-myc results in decreased homotypic cell aggregation and increased proliferative capacity of SCLC cells when nutrient conditions are adequate. We also find that c-myc contributes to apoptosis when cells are nutrient depleted, and flow cytometry suggests that this enhanced apoptosis is associated with a failure to halt cell cycling, consistent with the experience in other cell types. We previously found that protein kinase C-beta (PKC-beta) expression in NCI H209 (209) SCLC cells increases markedly with c-myc transfection (L. F. Barr et al., Cancer Res., 51: 5514-5519, 1991), and we hypothesized that PKC-beta may mediate some of the effects of c-myc in these cells. We test this hypothesis by transfection of rat PKC-beta 1 and bovine PKC-beta 2 isoforms into 209 cells before and after transfection with c-myc. PKC-beta 1 transfection has no effect on these cells. However, PKC-beta 2 expression has distinct phenotypic consequences. In the parental cells, PKC-beta 2 expression results in increased homotypic cell aggregation and a prolonged doubling time. Furthermore, PKC-beta 2 expression increases the fraction of these cells in G0-G1. In the cells which express a transfected c-myc gene, PKC-beta 2 expression improves the survival of cells in low serum by decreasing myc-induced apoptosis. This effect was associated with, and may be mediated by, a selection for cells in the G0-G1 fraction. We postulate that transfection of c-myc into SCLC cells may select for those expressing the PKC-beta 2 gene because this signal transduction event protects against myc-induced apoptosis.

The c-myc oncogene is frequently amplified in cells grown from lung tumors and has been linked to the malignancy of these cancers. In support of this, c-myc transfection enhances the in vitro proliferation and soft agar cloning of human small cell lung cancer (SCLC) cells. In this study, we surprisingly found that c-myc expression suppressed the formation of tumors by SCLC cells in athymic nude mice. c-myc expression down-regulated the protein and transcript for vascular endothelial growth factor (VEGF) in these SCLC cells, as well as VEGF transcript in rat fibroblasts manipulated for c-myc expression and in liver cells of c-myc-transgenic mice. Finally, bivariate and multivariate analyses demonstrated that the probability of tumor formation from lung cancer cell lines was negatively correlated with the relative expression of c-Myc, positively correlated with the relative expression of VEGF, and that the latent time to tumor formation was increased by the expression of c-Myc and decreased by the expression of VEGF. We hypothesize that, for lung cancer cells, c-Myc suppresses the formation of tumors in vivo by down-regulating VEGF, and that the amplification of c-myc seen in cells grown from lung tumors with a poor prognosis is an artifact of selection for growth in vitro.

Tavernier, A., Campbell. S. E. & Golubic, S. 1992 07 15: A complex marine shallow-water boring trace: Dendrorete balani n. ichnogen. et ichnospec. Exceptionally well-preserved reticulate boring patterns in Pliocene barnacle shells are described as a new ichnogenus and species. The organism that formed these borings is a presumed phototroph, possibly belonging to green algae. The principles and needs of a taxonomic treatment of microborings as trace fossils in relation to that of their makers are discussed. It is suggested that ichno-taxonomy be extended to include modern traces, when the taxonomic identity of the agent cannot be ascertained. Bio-erosion, endoliths, ichnofossils, marine, microborings, shallow marine, shells.