Mauro Rongioletti

Microplastics are particles smaller than five millimeters deriving from the degradation of plastic objects present in the environment. Microplastics can move from the environment to living organisms, including mammals. In this study, six human placentas, collected from consenting women with physiological pregnancies, were analyzed by Raman Microspectroscopy to evaluate the presence of microplastics. In total, 12 microplastic fragments (ranging from 5 to 10 μm in size), with spheric or irregular shape were found in 4 placentas (5 in the fetal side, 4 in the maternal side and 3 in the chorioamniotic membranes); all microplastics particles were characterized in terms of morphology and chemical composition. All of them were pigmented; three were identified as stained polypropylene a thermoplastic polymer, while for the other nine it was possible to identify only the pigments, which were all used for man-made coatings, paints, adhesives, plasters, finger paints, polymers and cosmetics and personal care products.

Coronavirus disease-2019 (COVID-19) pandemic continues to threaten patients, societies, and economic and healthcare systems around the world. Like many other diseases, the host immune system determines the progress of COVID-19 and fatality. Modulation of inflammatory response and cytokine production using immunonutrition is a novel concept that has been applied to other diseases as well. Zinc, one of the anti-inflammatory and antioxidant micronutrient found in food with well-established role in immunity, is currently being used in some clinical trials against COVID-19. This review integrates the contemporary studies of role of zinc in antiviral immunity along with discussing its potential role against COVID-19, and ongoing COVID-19 clinical trials using zinc.

Copper homeostasis is strictly regulated by protein transporters and chaperones, to allow its correct distribution and avoid uncontrolled redox reactions. Several studies address copper as involved in cancer development and spreading (epithelial to mesenchymal transition, angiogenesis). However, being endogenous and displaying a tremendous potential to generate free radicals, copper is a perfect candidate, once opportunely complexed, to be used as a drug in cancer therapy with low adverse effects. Copper ions can be modulated by the organic counterpart, after complexed to their metalcore, either in redox potential or geometry and consequently reactivity. During the last four decades, many copper complexes were studied regarding their reactivity toward cancer cells, and many of them could be a drug choice for phase II and III in cancer therapy. Also, there is promising evidence of using 64Cu in nanoparticles as radiopharmaceuticals for both positron emission tomography (PET) imaging and treatment of hypoxic tumors. However, few compounds have gone beyond testing in animal models, and none of them got the status of a drug for cancer chemotherapy. The main challenge is their solubility in physiological buffers and their different and non-predictable mechanism of action. Moreover, it is difficult to rationalize a structure-based activity for drug design and delivery. In this review, we describe the role of copper in cancer, the effects of copper-complexes on tumor cell death mechanisms, and point to the new copper complexes applicable as drugs, suggesting that they may represent at least one component of a multi-action combination in cancer therapy.

Microplastics (MPs) are defined as plastic particles smaller than 5 mm. They have been found almost everywhere they have been searched for and recent discoveries have also demonstrated their presence in human placenta, blood, meconium, and breastmilk, but their location and toxicity to humans have not been reported to date. The aim of this study was twofold: 1. To locate MPs within the intra/extracellular compartment in human placenta. 2. To understand whether their presence and location are associated with possible structural changes of cell organelles. Using variable pressure scanning electron microscopy and transmission electron microscopy, MPs have been localized in ten human placentas. In this study, we demonstrated for the first time the presence and localization in the cellular compartment of fragments compatible with MPs in the human placenta and we hypothesized a possible correlation between their presence and important ultrastructural alterations of some intracytoplasmic organelles (mitochondria and endoplasmic reticulum). These alterations have never been reported in normal healthy term pregnancies until today. They could be the result of a prolonged attempt to remove and destroy the plastic particles inside the placental tissue. The presence of virtually indestructible particles in term human placenta could contribute to the activation of pathological traits, such as oxidative stress, apoptosis, and inflammation, characteristic of metabolic disorders underlying obesity, diabetes, and metabolic syndrome and partially accounting for the recent epidemic of non-communicable diseases.

Abnormal handling of copper is the cause of Wilson disease (WD), a rare disorder typified by increased levels in plasma copper not-bound to ceruloplasmin (nCp-Cu, also known as ‘free’ copper). In Alzheimer’s disease (AD), meta-analyses show that copper decreases in brain but increases in serum, due to the nCp Cu component increase. Despite the similarities, a direct comparison of copper biological status in the two diseases has never been carried out. To fill this gap, we evaluated serum copper, ceruloplasmin, nCp-Cu and Cu:Cp in 385 CE and 336 healthy controls previously investigated that were compared with 9 newly diagnosed WD patients. We then assessed 24 h copper urinary excretion in 24 WD patients under D-penicillamine (D-pen) treatment and in 35 healthy controls, and compared results with those of AD patients participating to a D-pen phase II clinical trial previously published. After adjusting for sex and age, serum nCp-Cu and Cu:Cp resulted higher in AD and in WD than in healthy controls (both p < 0.001). While nCp-Cu was similar between AD and WD, Cu:Cp was higher in WD (p < 0.016). 24 h urinary copper excretion in AD patients (12.05 μg/day) was higher than in healthy controls (4.82 μg/day; p < 0.001). 77.8% of the AD patients under D-pen treatment had a 24 h urinary excretion higher than 200 μg/day, suggestive of a failure of copper control. This study provides new insight into the pathophysiology of copper homeostasis in AD, showing a failure of copper control and the Cu:Cp ratio as an eligible marker.

The dynamics of essential metals such as Copper (Cu) and Zinc (Zn) may be associated with the novel coronavirus disease 2019 (COVID-19) that has spread across the globe.The aim of this study is to investigate the relationship between serum levels of Cu and Zn, as well as the Cu:Zn ratio in the acute phase of COVID-19 along with the assessment of their connection to other laboratory parameters (hematological, biochemical, hemostatic).Serum levels of Cu and Zn were measured by atomic absorption spectrometry in 75 patients in the acute COVID-19 phase and were compared with those of 22 COVID-19 patients evaluated three months after the acute phase of the disease ('non-acute' group) and with those of 68 healthy individuals.In comparison with both the non-acute patients and the healthy controls, the acute patients had lower levels of hemoglobulin and albumin, and higher levels of glucose, creatinine, liver transaminases, C-reactive protein (CRP), and higher values of the neutrophils to lymphocytes ratio (NLR) at the hospital admission. They also exhibited increased levels of Cu and decreased of Zn, well represented by the Cu:Zn ratio which was higher in the acute patients than in both non-acute patients (p = 0.001) and healthy controls (p < 0.001), with no statistical difference between the last two groups. The Cu:Zn ratio (log scale) positively correlated with CRP (log scale; r = 0.581, p < 0.001) and NLR (r = 0.436, p = 0.003).Current results demonstrate that abnormal dynamics of Cu and Zn levels in serum occur early during the course of COVID-19 disease, and are mainly associated with the inflammation response.

Alzheimer's disease (AD) is a type of dementia whose cause is incompletely defined. Copper (Cu) involvement in AD etiology was confirmed by a meta-analysis on about 6000 participants, showing that Cu levels were decreased in AD brain specimens, while Cu and non-bound ceruloplasmin Cu (non-Cp Cu) levels were increased in serum/plasma samples. Non-Cp Cu was advocated as a stratification add-on biomarker of a Cu subtype of AD (CuAD subtype). To further circumstantiate this concept, we evaluated non-Cp Cu reliability in classifying subtypes of AD based on the characterization of the cognitive profile. The stratification of the AD patients into normal AD (non-Cp Cu ≤ 1.6 µmol/L) and CuAD (non-Cp Cu > 1.6 µmol/L) showed a significant difference in executive function outcomes, even though patients did not differ in disease duration and severity. Among the Cu-AD patients, a 76-year-old woman showed significantly abnormal levels in the Cu panel and underwent whole exome sequencing. The CuAD patient was detected with possessing the homozygous (c.1486T > C; p.(Ter496Argext*19) stop-loss variant in the RGS7 gene (MIM*602517), which encodes for Regulator of G Protein Signaling 7. Non-Cp Cu as an add-on test in the AD diagnostic pathway can provide relevant information about the underlying pathological processes in subtypes of AD and suggest specific therapeutic options.

Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide and a public health problem. Several clinical studies have shown that copper (Cu) is involved in carcinogenesis, possibly via cuproptosis, a new form of programmed cell death, but the conclusions from published reports are inconsistent. This study aimed at evaluating the potential of Cu dysregulation as a CRC susceptibility factor. In this systematic review and meta-analysis, we searched Cochrane Library, EBSCOhost, EMBASE, ProQuest, PubMed/MEDLINE, Scopus, and Web of Science for studies reporting serum Cu concentrations in CRC patients and controls from articles published till June 2023. The studies included reported measurements of serum/plasma/blood Cu levels. Meta-analyses were performed as well as study quality, heterogeneity, and small study effects were assessed. Based on a random effects model, summary standardized mean differences (SMDs) and the corresponding 95% confidence intervals (95% CIs) were applied to compare the levels of Cu between CRC patients and controls. 26 studies with a pooled total of9628 participants and 2578 CRC cases were included. The pooled SMD was equal to 0.85 (95% CIs -0.44; 2.14) showing that the CRC patients had higher mean Cu levels than the control subjects, but the difference was not significant (p=0.185) and the heterogeneity was very high, I2=97.9% (95% CIs: 97.5-98.3%; p<0.001). The pooled results were inconclusive, likely due to discordant results and inaccuracy in reporting data of some studies; further research is needed to establish whether Cu dysregulation might contribute to the CRC risk and whether it might reflect different CRC grades.

Oxidative status may play a role in chronic inflammation and neurodegeneration which are considered critical etiopathogenetic factors in Multiple Sclerosis (MS), both in the early phase of the disease and in the progressive one. The aim of this study is to explore oxidative status related to iron metabolism in peripheral blood of stable Relapsing-Remitting MS with low disability. We studied 60 Relapsing-Remitting MS patients (age 37.2 ± 9.06, EDSS median 1.0), and 40 healthy controls (age 40.3 ± 10.86). We measured total hydroperoxides (dROMs test) and Total Antioxidant Status (TAS), along with the iron metabolism biomarkers: Iron (Fe), ferritin (Ferr), transferrin (Tf), transferrin saturation (Tfsat), and ceruloplasmin (Cp) panel biomarkers [concentration (iCp) and enzymatic activity (eCp), copper (Cu), ceruloplasmin specific activity (eCp:iCp), copper to ceruloplasmin ratio (Cu:Cp), non-ceruloplasmin copper (nCp-Cu)]. We computed also the Cp:Tf ratio as an index of oxidative stress related to iron metabolism. We found lower TAS levels in MS patients than in healthy controls (CTRL) and normal reference level and higher dROMs and Cp:Tf ratio in MS than in healthy controls. Cp and Cu were higher in MS while biomarkers of iron metabolism were not different between patients and controls. Both in controls and MS, dROMs correlated with iCp (CTRL r = 0.821, p < 0.001; MS r = 0.775 p < 0.001) and eCp (CTRL r = 0.734, p < 0.001; MS r = 0.820 p < 0.001). Moreover, only in MS group iCp correlated negatively with Tfsat (r = -0.257, p = 0.047). Dividing MS patients in "untreated" group and "treated" group, we found a significant difference in Fe values [F(2, 97) = 10.136, p < 0.001]; in particular "MS untreated" showed higher mean values (mean = 114.5, SD = 39.37 μg/dL) than CTRL (mean 78.6, SD = 27.55 μg/dL p = 0.001) and "MS treated" (mean = 72.4, SD = 38.08 μg/dL; p < 0.001). Moreover, "MS untreated" showed significantly higher values of Cp:Tf (mean = 10.19, SD = 1.77∗10-2; p = 0.015), than CTRL (mean = 9.03, SD = 1.46 ∗10-2). These results suggest that chronic oxidative stress is relevant also in the remitting phase of the disease in patients with low disability and short disease duration. Therefore, treatment with antioxidants may be beneficial also in the early stage of the disease to preserve neuronal reserve.

Copper is an essential nutrient for plants, animals, and humans because it is an indispensable component of several essential proteins and either lack or excess are harmful to human health. Recent studies revealed that the breakdown of the regulation of copper homeostasis could be associated with Alzheimer’s disease (AD), the most common form of dementia. Copper accumulation occurs in human aging and is thought to increase the risk of AD for individuals with a susceptibility to copper exposure. This review reports that one of the leading causes of copper accumulation in the environment and the human food chain is its use in agriculture as a plant protection product against numerous diseases, especially in organic production. In the past two decades, some countries and the EU have invested in research to reduce the reliance on copper. However, no single alternative able to replace copper has been identified. We suggest that agroecological approaches are urgently needed to design crop protection strategies based on the complementary actions of the wide variety of crop protection tools for disease control.

Evidence indicates that patients with Alzheimer's dementia (AD) show signs of copper (Cu) dyshomeostasis. This study aimed at evaluating the potential of Cu dysregulation as an AD susceptibility factor. We performed a meta-analysis of 56 studies investigating Cu biomarkers in brain specimens (pooled total of 182 AD and 166 healthy controls, HC) and in serum/plasma (pooled total of 2929 AD and 3547 HC). We also completed a replication study of serum Cu biomarkers in 97 AD patients and 70 HC screened for rs732774 and rs1061472 ATP7B, the gene encoding for the Cu transporter ATPase7B. Our meta-analysis showed decreased Cu in AD brain specimens, increased Cu and nonbound ceruloplasmin (Non-Cp) Cu in serum/plasma samples, and unchanged ceruloplasmin. Serum/plasma Cu excess was associated with a three to fourfold increase in the risk of having AD. Our replication study confirmed meta-analysis results and showed that carriers of the ATP7B AG haplotype were significantly more frequent in the AD group. Overall, our study shows that AD patients fail to maintain a Cu metabolic balance and reveals the presence of a percentage of AD patients carrying ATP7B AG haplotype and presenting Non-Cp Cu excess, which suggest that a subset of AD subjects is prone to Cu imbalance. This AD subtype can be the target of precision medicine-based strategies tackling Cu dysregulation.

Reactive oxygen species (ROS) play a key role in the neurodegeneration processes. Increased oxidative stress damages lipids, proteins, and nucleic acids in brain tissue, and it is tied to the loss of biometal homeostasis. For this reason, attention has been focused on transition metals involved in several biochemical reactions producing ROS. Even though a bulk of evidence has uncovered the role of metals in the generation of the toxic pathways at the base of Alzheimer's disease (AD), this matter has been sidelined by the advent of the Amyloid Cascade Hypothesis. However, the link between metals and AD has been investigated in the last two decades, focusing on their local accumulation in brain areas known to be critical for AD. Recent evidence revealed a relation between iron and AD, particularly in relation to its capacity to increase the risk of the disease through ferroptosis. In this review, we briefly summarize the major points characterizing the function of iron in our body and highlight why, even though it is essential for our life, we have to monitor its dysfunction, particularly if we want to control our risk of AD.

Meta-analyses demonstrate copper involvement in Alzheimer's disease (AD), and the systemic ceruloplasmin status in relation to copper is an emerging issue. To deepen this matter, we evaluated levels of ceruloplasmin concentration, ceruloplasmin activity, ceruloplasmin specific activity (eCp/iCp), copper, non-ceruloplasmin copper iron, transferrin, the ceruloplasmin/transferrin ratio, and the APOE genotype in a sample of 84 AD patients and 58 healthy volunteers. From the univariate logistic analyses we found that ceruloplasmin concentration, eCp/iCp, copper, transferrin, the ceruloplasmin/transferrin ratio, and the APOE genotype were significantly associated with the probability of AD. In the multivariable logistic regression analysis, we selected the best subset of biological predictors by the forward stepwise procedure. The analysis showed a decrease of the risk of having AD for eCp/iCp (p = 0.001) and an increase of this risk for non-ceruloplasmin copper (p = 0.008), age (p = 0.001), and APOE-ɛ4 allele (p < 0.001). The estimated model showed a good power in discriminating AD patients from healthy controls (area under curve: 88% ; sensitivity: 66% ; specificity 93%). These data strength the breakdown of copper homeostasis and propose eCp/iCp as a reliable marker of ceruloplasmin status.

Dysfunction of the complex cerebral networks underlying wakefulness and awareness is responsible for Disorders of Consciousness (DoC). Traumatic Brain Injury (TBI) is a common cause of DoC, and it is responsible for a multi-dimensional pathological cascade that affects the proper functioning of the brainstem and brain consciousness pathways. Iron (Fe), Zinc (Zn), and Copper (Cu) have a role in the neurophysiology of both the ascending reticular activating system, a multi-neurotransmitter network located in the brainstem that is crucial for consciousness, and several brain regions. We aimed to summarize the role of these essential metals in TBI and its possible link with consciousness alterations. We found that TBI alters many neuronal molecular mechanisms involving essential metals, causing neurodegeneration, neural apoptosis, synaptic dysfunction, oxidative stress, and inflammation. This final pattern resembles that described for Alzheimer's disease (AD) and other neurological and psychiatric diseases. Furthermore, we found that amantadine, zolpidem, and transcranial direct current stimulation (tDCS)-the most used treatments for DoC recovery-seem to have an effect on essential metals-related pathways and that Zn might be a promising new therapeutic approach. This review summarizes the neurophysiology of essential metals in the brain structures of consciousness and focuses on the mechanisms underlying their imbalance following TBI, suggesting their possible role in DoC. The scenario supports further studies aimed at getting a deeper insight into metals' role in DoC, in order to evaluate metal-based drugs, such as metal complexes and metal chelating agents, as potential therapeutic options.

Type 1 diabetes (T1D) is a chronic condition in which the pancreas loses the ability to produce insulin due to an autoimmune destruction of the insulin producing beta cells in the pancreatic islets of Langerhans. Pathophysiological complications related to diabetes include micro and macrovascular disease, nephropathy, and neuropathy that can also be affected by environmental factors such as lifestyle and diet.The current study aimed to evaluate the serum levels of total copper, the copper-carrying protein, ceruloplasmin and nonceruloplasmin bound copper (nonceruloplasmin-Cu) and other essential and environmental metals and metalloids in subjects with T1D compared with healthy controls.A cohort of 63 subjects with T1D attending Diabetes Clinics at the University of Miami and 65 healthy control subjects was studied. Metals and metalloids were measured by inductively coupled plasma mass spectrometry.A main finding of this study was that total copper and ceruloplasmin levels were higher in persons with T1D compared to healthy controls. In comparison to other metals and clinical variables, elevated copper was the strongest factor associated with T1D resulting in a15-fold increased odds of having the disease per standard deviation increase.Our results suggest a metal and metalloid perturbation in T1D with a significant involvement of Copper dysfunction in the disease pathology, possibly linked to inflammatory processes.

Zinc therapy is normally utilized for treatment of Wilson disease (WD), an inherited condition that is characterized by increased levels of non-ceruloplasmin bound ('free') copper in serum and urine. A subset of patients with Alzheimer's disease (AD) or its prodromal form, known as Mild Cognitive Impairment (MCI), fail to maintain a normal copper metabolic balance and exhibit higher than normal values of non-ceruloplasmin copper. Zinc's action mechanism involves the induction of intestinal cell metallothionein, which blocks copper absorption from the intestinal tract, thus restoring physiological levels of non-ceruloplasmin copper in the body. On this basis, it is employed in WD. Zinc therapy has shown potential beneficial effects in preliminary AD clinical trials, even though the studies have missed their primary endpoints, since they have study design and other important weaknesses. Nevertheless, in the studied AD patients, zinc effectively decreased non-ceruloplasmin copper levels and showed potential for improved cognitive performances with no major side effects. This review discusses zinc therapy safety and the potential therapeutic effects that might be expected on a subset of individuals showing both cognitive complaints and signs of copper imbalance.

Background: Meta-analyses show that percentages of non-Cp-Cu—copper that is not bound to ceruloplasmin (also known as ‘free’ copper)—in serum are higher in Alzheimer’s disease (AD) patients. Genetic heterogeneity in AD patients stratified on the basis of non-Cp-Cu cut-off sustains the existence of a copper AD metabolic subtype. Non-Cp-Cu abnormalities correlated with alterations of electroencephalographic rhythms (EEG). Objective: We aimed to determine whether an EEG-derived brain cortical rhythm’s heterogeneity between two AD groups stratified on the basis of a copper marker. Method: We assessed levels of copper, ceruloplasmin, Non-Cp-Cu, and the APOE4 genotype in 67 AD patients and compared resting EEG-derived eLORETA cortical rhythms between AD groups stratified in terms of ‘Normal’ and ‘High’ non-Cp-Cu. Results: The High non-Cp-Cu group experienced a lower power in all bands (0.2-48 Hz) in the parietal cortices (p=0.019) and a more limited alpha band (8-13 Hz) power in the sensory lobes (temporal, occipital, and parietal p<0.05 consistently) than the Normal non-Cp-Cu AD group. When corrected for MMSE, the non-Cp-Cu levels correlated with a reduction of high-frequency brain activity (from high alpha to gamma, 10.5-48 Hz). Conclusion: This neurophysiological heterogeneity in EEG-derived brain cortical rhythms between the two AD groups sustains a copper AD metabolic subtype; Non-Cp-Cu is a marker of this copper AD. Keywords: Alzheimer's disease (AD) subtype, ceruloplasmin, cortical lobes, Electroencephalography (EEG), free copper.

GENERAL COMMENTARY article Front. Neurol., 25 September 2017Sec. Neurodegeneration Volume 8 - 2017 | https://doi.org/10.3389/fneur.2017.00503

Background: Meta-analyses show that copper non-bound to ceruloplasmin (non-Cp Cu, also known as ‘free’ copper) in serum is higher in a percentage of Alzheimer's disease (AD) patients. Genetic heterogeneity in AD patients stratified on the basis of non-Cp Cu cut-off sustains the existence of a copper AD metabolic subtype. Objective: In order to find evidence of the existence of a detectable metabolic subtype of AD associated to copper abnormalities, we explore the hypothesis of a neuroimaging pattern heterogeneity in an homogenous and well characterized AD population classified in two groups by the stratification of patients on the basis non-Cp Cu cut-off. Method: We assessed levels of copper, ceruloplasmin, non-Cp Cu, cerebrospinal levels of total Tau protein (h-tau), Thr 181 phosphorylated tau protein (P-tau) and β-amyloid 1-42, and APOE4 genotype in 66 AD patients and compared neuroimaging indices of a visual rating scale of cerebral atrophy and neurovascular burden in AD patients stratified in ‘Normal’ and ‘High’ non-Cp Cu groups. Results: The stratification for non-Cp Cu originated AD groups which did not differ for medial temporal lobe atrophy, periventricular hyperintensities, deeper hyperintensities (including frontal, parietooccipital and temporal white matter hyperintensities), infratentorial hyperintensities indices, while they differed for global atrophy. More specifically, AD patients within the high non-Cp Cu group had a less severe burden of global atrophy (p=0.042). Conclusion: This neuroimaging heterogeneity between AD groups is suggestive of the existence of a copper metabolic subtype of AD; non-Cp Cu appears a good marker of this copper AD. Keywords: Neuroimaging heterogeneity, copper not bound to ceruloplasmin (non-Cp Cu), Alzheimer's disease, free copper, global atrophy, copper.

Colorectal cancer (CRC) is a growing public health problem. Several clinical studies have shown a potentially protective effect of selenium (Se), but the reports are inconsistent. The objective of the study was to examine the evidence for relation between serum/tissue Se status and CRC.In this Systematic Review and Meta-Analysis, we searched Cochrane Library, EBSCOhost, EMBASE, ProQuest, PubMed/MEDLINE, Scopus, and Web of Science for studies reporting serum/plasma/whole blood/tissue Se concentrations in CRC patients and controls for articles published till August 2023. Meta-analysis was performed, and study quality, heterogeneity, and small study effects were assessed. Based on a random effects model, summary mean differences in serum levels of Se between CRC patients and healthy controls, and Se levels between malignant and matched non-malignant tissue specimens were assessed.After initial screening, a total of 24 studies (18 serum and 6 tissue studies) with a pooled total of 2640 participants were included in the meta-analysis. CRC patients had significantly lower serum Se levels than healthy controls, being the difference between the two equal to 3.73 µg/dl (95% CI: 6.85-0.61). However, the heterogeneity was very high, I2= 99% (p < 0.01). Our meta-analysis showed higher Se levels in CRC cancerous specimens than in matched healthy colon tissue: the increase was equal to 0.07 µg/g wet tissue weight (95% CI: 0.06-0.09; p= 0.02).CRC patients have lower serum and higher colon cancerous tissue Se levels. Some factors, such as Se levels in different tumor grades of CRC need to be further considered for a more conclusive association between Se levels and risk of CRC.

Evidence of copper's (Cu) involvement in Alzheimer's disease (AD) is available, but information on Cu involvement in microglia and astrocytes during the course of AD has yet to be structurally discussed. This review deals with this matter in an attempt to provide an updated discussion on the role of reactive glia challenged by excess labile Cu in a wide picture that embraces all the major processes identified as playing a role in toxicity induced by an imbalance of Cu in AD.

Non-coding RNAs (ncRNAs), including microRNAs (miRNAs), are key regulators of differentiation and development. In the cell, transcription factors regulate the production of miRNA in response to different external stimuli. Copper (Cu) is a heavy metal and an essential micronutrient with widespread industrial applications. It is involved in a number of vital biological processes encompassing respiration, blood cell line maturation, and immune responses. In recent years, the link between deregulation of miRNAs' functionality and the development of various pathologies as well as cardiovascular diseases (CVDs) has been extensively studied. Alzheimer's disease (AD) is the most common cause of dementia in the elderly with a complex disease etiology, and its link with Cu abnormalities is being increasingly studied. A direct interaction between COMMD1, a regulator of the Cu pathway, and hypoxia-inducible factor (HIF) HIF-1a does exist in ischemic injury, but little information has been collected on the role of Cu in hypoxia associated with AD thus far. The current review deals with this matter in an attempt to structurally discuss the link between miRNA expression and Cu dysregulation in AD and CVDs.

Summary Asthma and allergies are characterized by variable and subjective symptoms influenced by many genes, molecular mechanisms and environmental factors. The presence of inflammation and oxidative stress in the airways are important biochemical features of asthma and respiratory allergies. Glutathione S‐transferase ( GST s) enzymes play an important role in cellular protection against inflammation, and functional genetic polymorphisms in GST genes show a significant association with asthma and allergy risk. Specifically, our previous study on asthmatic children highlighted GSTA 1 and GSTO 2 as novel susceptibility loci for asthma. In the present study we focused our attention on GSTA 1*‐69C/T (rs3957357) and GSTO 2*N142D (rs156697) polymorphisms to confirm our previous results in an independent adult study population and to clarify whether GSTA 1 and GSTO 2 gene polymorphisms are involved in a non‐discriminative pathway towards asthma and respiratory allergy. To accomplish this, we recruited 103 patients with respiratory allergies, 199 patients with asthma and 200 healthy controls. Genomic DNA extracted from buccal cells was screened for GSTA 1*‐69C/T and GSTO 2*N142D single nucleotide polymorphisms. The GSTA 1*‐69T and GSTO 2*D142 variants are both associated with a significantly increased risk of asthma, whereas only GSTA 1*‐69C/T is significantly associated with allergies. These outcomes confirm the involvement of GSTO 2 loci in asthma and suggest that GSTA 1 is a common risk factor for asthma and allergies.

Alzheimer's Disease (AD) is a type of dementia very common in the elderly. A growing body of recent evidence has linked AD pathogenesis to Copper (Cu) dysmetabolism in the body. In fact, a subset of patients affected either by AD or by its prodromal form known as Mild Cognitive Impairment (MCI) have been observed to be unable to maintain a proper balance of Cu metabolism and distribution and are characterized by the presence in their serum of increased levels of Cu not bound to ceruloplasmin (non-ceruloplasmin Cu). Since serum non-ceruloplasmin Cu is a biomarker of Wilson's Disease (WD), a well-known condition of Cu-driven toxicosis, in this review, we propose that in close analogy with WD, the assessment of non-ceruloplasmin Cu levels can be exploited as a cost-effective stratification and susceptibility/risk biomarker for the identification of some AD/MCI individuals. The approach can also be used as an eligibility criterion for clinical trials aiming at investigating Cu-related interventions against AD/MCI.

Alzheimer’s disease is a progressive neurodegenerative disorder that eventually leads the affected patients to die. The appearance of senile plaques in the brains of Alzheimer’s patients is known as a main symptom of this disease. The plaques consist of different components, and according to numerous reports, their main components include beta-amyloid peptide and transition metals such as copper. In this disease, metal dyshomeostasis leads the number of copper ions to simultaneously increase in the plaques and decrease in neurons. Copper ions are essential for proper brain functioning, and one of the possible mechanisms of neuronal death in Alzheimer’s disease is the copper depletion of neurons. However, the reason for the copper depletion is as yet unknown. Based on the available evidence, we suggest two possible reasons: the first is copper released from neurons (along with beta-amyloid peptides), which is deposited outside the neurons, and the second is the uptake of copper ions by activated microglia.

The clinical course after ischemic stroke can vary considerably despite similar lesions and clinical status at the onset of symptoms, suggesting that individual factors modulate clinical recovery. Here, we sought to test the working hypothesis that elevated copper values provide prognostic information, and specifically predict worse clinical recovery. We further sought to support previous findings regarding metal metabolism in acute stroke. We assessed total antioxidant status, oxidative stress factors (peroxides) and metal metabolism markers (iron, copper, ceruloplasmin concentration and activity, ferritin, and transferrin) in the acute phase (2-10 days from symptom onset) in 30 patients affected by unilateral middle cerebral artery (MCA) stroke. A longitudinal assessment of clinical deficit was performed in the acute and stabilized phases (typically 6 months post-stroke) using the National Institutes of Health Stroke Scale (NIHSS). In identifying recovery-related factors, we considered effective recovery (ER), calculated as the ratio between actual NIHSS recovery and the total potential recovery. This allows an estimation of the actual recovery adjusted for the patient's initial condition. In the acute phase, clinical severity was correlated with increased peroxide concentrations, and lower iron levels. Less successful clinical recovery was correlated with increased acute copper levels, which entered a multiple regression model that explained 24% of ER variance. These pilot data suggest that, in the acute phase of an ischemic stroke, copper may provide useful information about clinical recovery.

Background: Life expectancy is increasing all over the world, although neurodegenerative disorders might drastically affect the individual activity of aged people. Of those, Alzheimer’s Disease (AD) is one of the most social-cost age-linked diseases of industrialized countries. To date, retinal diseases seem to be more common in the developing world and characterize principally aged people. Agerelated Macular Degeneration (AMD) is a late-onset, neurodegenerative retinal disease that shares several clinical and pathological features with AD, including stress stimuli such as oxidative stress, inflammation and amyloid formations. Method: In both diseases, the detrimental intra/extra-cellular deposits have many similarities. Aging, hypercholesterolemia, hypertension, obesity, arteriosclerosis and smoking are risk factors to develop both diseases. Cellular aging routes have similar organelle and signaling patterns in retina and brain. The possibility to find out new research strategies represent a step forward to disclose potential treatment for both of them. Essential trace metals play critical roles in both physiological and pathological condition of retina, optic nerve and brain, by influencing metabolic processes chiefly upon complex multifactorial pathogenesis. Conclusion: Hence, this review addresses current knowledge about some up-to-date investigated essential trace metals associated with AD and AMD. Changes in the levels of systemic and ocular fluid essential metals might reflect the early stages of AMD, possibly disclosing neurodegeneration pathways shared with AD, which might open to potential early detection.

Plasma homocysteine levels depend in part on the molecular nature of the methylenetetrahydrofolate reductase (MTHFR) and on blood folate intake. Little has been reported on platelet counts in the presence of hyperhomocysteinemia and MTHFR polymorphisms, with the exception of delayed platelet recovery in homozygous MTHFR C677T subjects after treatment with methotrexate for ovarian cancer. The aim of this investigation was to evaluate the possibility of a link between the platelet count and plasma homocysteine levels in different MTHFR variants in 165 female patients. Determinations of plasma homocysteine levels were by ELISA and of MTHFR polymorphisms (A1298C and C677T) were by inverse hybridization. Serum P- and E-selectin concentrations were obtained by ELISA. An inverse correlation (R=-0.88, P<0.001) was observed between blood platelet counts and plasma homocysteine levels in the women homozygous for MTHFR C677T. This correlation did not depend on pregnancy or other variables reported. Serum concentrations of sE- and sP-selectin, markers of endothelial and platelet activation, were significantly and positively correlated with homocysteine levels. These findings suggest that homocysteine affects platelet numbers in women with MTHFR C677T possibly consequent to endothelial and platelet activation.

Iatrogenic anaemia caused by repeated blood sampling to monitor laboratory parameters can contribute, particularly in neonates, to the need for transfusion. "Point of care" laboratory equipment uses smaller amounts of blood for analytic determinations and could, therefore, help to prevent secondary anaemia. In this study we compared the results of haematological parameters measured using a standard laboratory method and using a "point of care" micromethod, with the aim of validating the use of this latter method in clinical practice in neonatology.One hundred and fifty venous or capillary blood samples were taken from full-term or premature neonates 2-4 hours or 48 hours after birth. Each sample was processed by a standard haematology analyser and another micromethod instrument. Bland-Altman plots were constructed for each parameter and intra-class coefficients of correlation were calculated in order to evaluate the concordance between the two analysers.The concordance between the data obtained with the two analysers, expressed as the intra-class correlation, was 0.98 for white blood cell count, 0.97 for haemoglobin concentration, 0.96 for haematocrit, 0.95 for mean red cell volume and 0.98 for platelet count. The micromethod produced overestimated mean values for the leucocyte count (+1.27; p<0.001), haematocrit (+1.80; p<0.001) and platelet count (+13.55; p<0.001).Overall, the concordance between the values obtained with the two analysers was high for each of the parameters taken into consideration. In the case of haemoglobin and leucocytes, give the high intra-class correlation and lack of systematic overestimation of one method over another, the micromethod guarantees a correct evaluation; however, despite the high intra-class correlations for platelet counts, the systemic error seems to suggest that the micromethod cannot guarantee an appropriate evaluation of this parameter.

Meta-analyses show copper dyshomeostasis in Alzheimer’s disease. However, a study evaluating copper changes in other neurodegenerative forms of dementia has not yet been performed. In this study, we assessed copper, ceruloplasmin, copper not bound to ceruloplasmin, and copper to ceruloplasmin ratio in 85 patients affected by frontotemporal lobar degeneration (FTLD) and 55 healthy controls. Data were analyzed through multivariate ANOVA models taking into account age and sex as covariates and the stratification for FTLD variants, after calculating power analysis to ensure the reliability of the conclusions drawn. The study revealed no difference between the groups.

Background: The compilation of a list of genetic modifiers in Alzheimer’s disease (AD) is an open research field. The GABAergic system is affected in several neurological disorders but its role in AD is largely understudied. Objective/Methods: As an explorative study, we considered variants in genes of GABA catabolism (ABAT, ALDH5A1, AKR7A2), and APOE in 300 Italian patients and 299 controls. We introduce a recent multivariate method to take into account the individual APOE genotype, thus controlling for the effect of the discrepant allele distributions in cases versus controls. We add a genotype-phenotype analysis based on age at onset and the Mini-Mental State Evaluation score. Results: On the background of strongly divergent APOE allele distributions in AD versus controls, two genotypic interactions that represented a subtle but significant peculiarity of the AD cohort emerged. The first is between ABAT and APOE, and the second between some ALDH5A1 genotypes and APOE. Decreased SSADH activity is predicted in AD carriers of APOE ɛ4, representing an additional suggestion for increased oxidative damage. Conclusion: We identified a difference between AD and controls, not in a shift of the allele frequencies at genes of the GABA catabolism pathway, but rather in gene interactions peculiar of the AD cohort. The emerging view is that of a multifactorial contribution to the disease, with a main risk factor (APOE), and additional contributions by the variants here considered. We consider genes of the GABA degradation pathway good candidates as modifiers of AD, contributing to energy impairment in AD brain.

Abnormal complete blood count (CBC) and high plasma C-reactive protein (CRP) are associated with neonatal infections and could be helpful in the diagnosis of neonatal sepsis and to monitor the antibiotic treatment.The aim of this work is to evaluate and compare the performance of a bedside analyzer for blood count and C-reactive protein (CRP) with a conventional analyzer in a neonatal population.150 capillary or venous blood samples of term and preterm newborns were processed on an ABX-MicrosCRP200 analyzer and on a SysmexXE2100 (conventional hematology analyzer) for CBC, leukocyte differential, reticulocytes, and nucleated red blood cells (NRBC); high-sensitivity CRP (hs-CRP) was performed on a ModularPE. The differences between complete blood count and CRP were regressed against their means and assessed by means of intra-class-correlation.The intra-class-correlation for white blood cell (WBC) was 0.98, for hemoglobin 0.97, for hematocrit 0.96, for mean corpuscular volume 0.95, and for platelet 0.98. ABX-MicrosCRP200 overestimated the WBC (+1.27 x 10(3)/microL; p < 0.001), hematocrit (+1.80%; p < 0.001), and platelet (+13.55 x 10(3)/microL; p < 0.001). The intra-class-correlation for CRP was high (0.97), without systematic difference between the two values (p = 0.64).The agreement between the two methods was high for both tests. However, the SD of the difference for WBC and platelet could be clinically important in leukopenic or thrombocytopenic newborns.

The placenta is a crucial interface between the fetus and the maternal environment. It allows for nutrient absorption, thermal regulation, waste elimination, and gas exchange through the mother's blood supply. Furthermore, the placenta determines important adjustments and epigenetic modifications that can change the phenotypic expression of the individual even long after birth. Polyethylene glycol (PEG) is a polyether compound derived from petroleum with many applications, from medicine to industrial manufacturing. In this study, for the first time, an integration of ultra-high-performance liquid chromatography (UHPLC) coupled with mass spectrometry (MS) was used to detect suites of PEG compounds in human placenta samples, collected from 12 placentas, originating from physiological pregnancy. In 10 placentas, we identified fragments of PEG in both chorioamniotic membranes and placental cotyledons, for a total of 36 samples.

Summary paragraph Microplastics are particles smaller than five millimetres obtained from the degradation of plastic objects abandoned in the environment. Microplastics can move from the environment to living organisms and, in fact, they have been found in fishes and mammals. Six human placentas, prospectively collected from consenting women with uneventful pregnancies, were analyzed by Raman Microspectroscopy to evaluate the presence of microparticles. Detected microparticles were characterized in terms of morphology and chemical composition. 12 microparticles, ranging from 5 to 10 μm in size, were found in 4 out of 6 placentas: 5 in the foetal side, 4 in the maternal side and 3 in the chorioamniotic membranes. All the analyzed microparticles were pigmented: three of them were identified as stained polypropylene, while for the other nine it was possible to identify only the pigments, which are all used for man-made coatings, paints and dyes. Here we show, for the first time, the presence of microparticles and microplastics in human placenta. This sheds new light on the impact of plastic on human health. Microparticles and microplastics in the placenta, together with the endocrine disruptors transported by them, could have long-term effects on human health.

Frontotemporal lobar degeneration (FTLD) is a progressive neurodegenerative syndrome. Defects of copper (Cu) and iron (Fe) homeostasis are involved in the development of several neurodegenerative diseases and their homeostasis is interconnected by the Cu-protein ceruloplasmin (Cp), responsible for Fe oxidative state. In this study we assessed Fe, transferrin (Trf), ferritin, Cp specific activity (eCp/iCp), Cp/Trf ratio, and Trf saturation in 60 FTLD patients and 43 healthy controls, and discussed the results in relation to Cu homeostasis. The significant decrease of the eCp/iCp in the FTLD patients supports the involvement of Fe imbalance in the onset and progression of FTLD.

Wolfram Syndrome 1 (WS1) has been characterized on the basis of mutation in the WFS1 gene encoding a calcium storage wolframin endoplasmatic reticulum transmembrane glycoprotein.We observed a WS 10-years old female subject, with Type 1 diabetes-mellitus (DM), that had compound heterozygous WSF1 mutations but without other symptoms generally observed in WS subjects, such as optic atrophy or neurodegeneration.Decreased copper, ceruloplasmin, and transferrin levels, pointing to a copper deficiency, were associated with a new c.18703A>G mutation in the ATP7B gene, while lower calcium levels were associated with WSF1 mutations. An omega-3 fatty acids therapy was administrated to the subject in the attempt to ameliorate diabetes symptoms, restored copper deficiency, and normal calcium levels.This specific case report provides new insights into the potential interplay of ATP7B mutation in shaping a milder WS clinical picture.

Copper (Cu) is an essential trace metal, and either a Cu deficiency or excess is life-threatening. Recent studies have indicated that an imbalance of Cu levels and a high burden of oxidative stress are contributors of Alzheimer's disease (AD), the main cause of dementia in the elderly. AD Cu imbalance can be described as a reduction of functional Cu bound to proteins and a shift of the metal to the non-ceruloplasmin (non-Cp) Cu pool, prone to oxidative stress. The shift/displacement of the metal from bound to labile toxic Cu in the brain can explain mitochondrial deficits, energy depletion, protein misfolding of amyloid beta, and other proteins associated to dementia. We suggest a diet-gene interplay at the basis of the “CuAD,” a subset of AD patients typified by higher-than-normal non-Cp Cu. A description of the Cu-regulating pathways and suggestions for a low-copper diet to improve preventive strategies are reported herein.

The placenta is a crucial interface between the fetus and the maternal environment. It allows for nutrient absorption, thermal regulation, waste elimination and gas exchange through the mother's blood supply. Furthermore, the placenta determines important adjustments and epigenetic modifications that can change the phenotypic expression of the individual even long after birth.Polyethylene glycol (PEG) is a polyether compound derived from petroleum with many applications, from medicine to industrial manufacturing. In this study, for the first time, a combination of UHPLC followed by Mass Spectrometry was used to detect suites of polyethylene glycol (PEG) compounds in human placenta samples, collected from 12 placentas, originating from physiological pregnancy. In all but two placentas, we identified fragments of PEG in chorioamniotic membranes, and in placental cotyledons, for a total of 36 samples.

15 Frequency of mutations of the cystic fibrosis transmembrane regulator (CFTR) gene in a cohort of consecutive patients candidate for assisted reproductive techniques M. Rongioletti1, F. Papa1, M.B. Majolini1, C. Vaccarella1, I. Simonelli2, A. Luciano1, P. Pasqualetti2, G.M. Liumbruno1. 1“San Giovanni Calibita” Fatebenefratelli Hospital, Clinical Pathology Laboratory, Molecular Biology Unit, Rome, Italy; 2Fatebenefratelli Association for the Research, Medical Statistics & Information Technology, Rome, Italy

Copper is an essential micronutrient, catalyst, or component of many metalloproteins and enzymes in physiological cellular functioning. Inborn error of copper excretion is the cause of Wilson disease (WD), a rare autosomal recessive disorder caused by mutations of the ATP7B gene, typified by increased levels in plasma copper not bound to ceruloplasmin (nCp-Cu, also known as “free” copper). The expansion of this plasma copper component is toxic since it can cross the blood–brain barrier and increase the labile pool of the metal in the brain. Meta-analyses have shown that nCp-Cu component increases also in Alzheimer disease (AD), the most common form of dementia, caused by the complex interaction and still mostly unknown factors. A recent study provides new insight into the pathophysiology of copper homeostasis in AD, likely originating from the same WD altered pathway but less severely damaged, being ATP7B variants associated with the increase of susceptibility for AD. Keeping clear the different etiology of the two diseases, we have posited the hypothesis that the AD typified by failure of copper control may represent a heterozygote form of WD.

Objectives: This study compare the performance of NucliSENS EasyQ test (bioMerieux), a RT-PCR based method, or HPV OncoTect (IncellDx) a flow cytometry-FISH based method, in the detection of the E6/E7 mRNA expression of hrHPV. Moreover, we investigated the potential role, for the detection of high-grade lesions, of the HPV OncoTect …

The purpose of this study was to compare the performance in the detection of the E6/E7 mRNA expression of hrHPV, using the NucliSENS EasyQ test (bioMerieux), a RT-PCR based method, or HPV OncoTect (IncellDx) a test based on the flow cytometry-FISH method. Moreover, we investigated the potential role, for the detection of high grade lesions, …

Objectives: Meta-analyses show that nonbound ceruloplasmin copper (Non-Cp-Cu, also known as “free” or labile copper) in serum is higher in patients with Alzheimer disease (AD). It was demonstrated that ATP7B gene associates with AD, and being carriers of some ATP7B variants accounted for a large proportion of serum Non-Cp-Cu levels, …

Cystic fibrosis (CF) is one of the most frequently diagnosed autosomal-recessive diseases in the Caucasian population. Screening for cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations, including poly T, is strongly recommended in infertile couples planning a pregnancy by assisted reproductive technology (ART). This study evaluated the performance of the new Nanochip …

Infertility affects about 15% of couples in Italy. Cystic fibrosis transmembrane conductance regulator (CFTR) is the gene causing the most common autosomal recessive disease in Caucasians, affecting one in 2,550 live birds. Men affected are usually infertile and asymptomatic heterozygote. The prevalence of the asymptomatic heterozygote is one to 27 individuals in Italy. The 5T allele in intron 8 (IVS8-poliT) of the CFTR gene produce an abnormally low levels of the CFTR protein, since it is associated with a higher proportion of mRNA transcripts lacking exon 9, with respect to the other two alleles 7T and 9T. To report results of a routine genetic screening and prevalence of CFTR mutations and 5T allele in a population of infertile patients who were candidates for assisted reproduction technique (ART) in the hospital San Giovanni Calibita, Fatebenefratelli, Isola Tiberina, Rome, Italy. A total of 3,746 couples and an additional 357 individuals referring to the Fatebenefratelli Hospital were analyzed through the CFTR INNO-LiPA Amplification kit (Innogenetics, Belgium), which detects 57 CFTR gene mutations and IVS8-poliT polymorphism, including both general and Italian regional strips. Among the screened couples, 9.2% were composed by at least a carrier of a CTFR mutation. Considering all 7,849 individuals, we found heterozygosis in one to 22 subjects. Compound heterozygosis was found in three individuals. Eight mutations were detected exclusively in men, and nine in women. 9T allele in ISV8-poliT was found in 65% of carriers of at least a CFTR mutation vs 20% of subjects with no mutation (P < .001). Conversely, 7T allele was found in 98% vs 90% (P < .001). Our results reinforce the relevance of an accurate determination of mutations in the CFTR gene, including the ISV8-poliT, in order to inform the couple of their carrier risk and the possibility of having an affected child.

Failure of copper control is the cause of Wilson disease (WD), a rare autosomal recessive disorder typified by increased levels of copper nonbound to ceruloplasmin (non-Cp Cu) and caused by mutations in the ATP7B gene, a copper transporter at the trans-Golgi network in the hepatocyte. Wilson disease has a broad spectrum of symptom presentation. About one-third of cases present with liver disease, another third with neurological manifestations, and the last third presents initially psychiatric and behavioral abnormalities. Based on mass screening studies, data about WD prevalence have been recently questioned, suggesting that WD might be much more common than estimated in the past (1984), passing from 1:30,000 to 1:3,000 or even 1:1,500 cases. Misdiagnosis for those cases with a psychiatric primary presentation can be suspected. To this aim, in September 2017, we started a project aimed at screening copper anomalies in patients with psychiatric disorders. In March 2017, we recruited 43 consecutive subjects with psychiatric disorders (anxiety disorder, mood disorder [major depression, bipolar depression], psychotic disorder [schizophrenia and delusional disorder], personality disorder) and compared them with 39 healthy control subjects for copper (Cu); ceruloplasmin concentration (iCp), activity (eCp), and specific activity (eCp/iCp); Cu:Cp ratio; iron; ferritin; and transferrin. Psychiatric clinical scale scores, medication, and routine lab tests were also collected. A next-generation sequencing of the ATP7B gene for the detection of WD mutations has been carried out to confirm a WD diagnosis. Recruitment is still ongoing. Our preliminary results show that the mean value of Cu (P < .05) and Cp (P < .005) in patients with psychiatric disorders is lower than that of healthy subjects. Moreover, 18% and 40% of patients have values lower than normal reference range for iCp and Cu, respectively. Among these subjects, WD patients with presentation of psychiatric symptoms could be hidden.

Copper failure is the cause of Wilson disease (WD), a rare disorder typified by increased levels of copper nonbound to ceruloplasmin (non-Cp Cu) and caused by mutations in the ATP7B gene, a copper transporter at the trans-Golgi network of the hepatocyte. Non-Cp Cu increases also in Alzheimer disease (AD), a multifactorial disorder and the main form of dementia in the elderly. Recent meta-analyses indicate reduced levels of copper in the brain, increased levels in the serum, and the expansion of a pool of non-Cp Cu, which typifies a subgroup of AD. However, a study evaluating copper changes in other neurodegenerative forms of dementia has not yet been performed. In this study, we assessed copper, ceruloplasmin, copper not bound to ceruloplasmin, and copper to ceruloplasmin ratio in 85 patients affected by frontotemporal lobar degeneration (FTLD), 60 AD patients, and 55 healthy controls. Data were analyzed through multivariate ANOVA models, taking into account age and sex as covariates and the stratification for FTLD clinical variants, after calculating power analysis to ensure the reliability of the conclusions drawn. A next-generation sequencing (NGS) study to search for specific ATP7B AD mutations on patients typified by increased values of non-Cp Cu has been carried out. The study revealed no difference between FTLD and healthy controls, while AD had increased values of non-Cp Cu. In the NGS study, we expect to detect a difference in the biochemical variables among genetic groups of at least 60%. With such expected changes, the size n = 60 is sufficient to detect a difference between genetic groups with a power of higher than 90% (0<0.05). Excess non-Cp Cu is not a common signature of dementia but it appears specific for AD.

Copper failure is the cause of Wilson disease (WD), a rare disorder typified by increased levels of copper nonbound to ceruloplasmin (non-Cp Cu) and copper-ceruloplasmin ratio (Cu:Cp). Meta-analyses also show copper abnormalities in Alzheimer disease (AD); however, a direct comparison of copper biological status in the two diseases has never been carried out. An investigation of copper status in serum (copper, ceruloplasmin, non-Cp Cu and Cu:Cp, adjusting for sex and age) and a 24-hour copper urine assessment of patients with AD, WD (nine WD patients at baseline and 24 WD patients under D-penicillamine [D-pen] treatment)], and healthy controls (35 healthy controls) was carried out. In a sample of 385 AD cases and 336 healthy controls investigated in previous studies, non-Cp Cu and Cu:Cp were higher in AD than healthy controls (both P < .001), but lower than in WD patients at baseline. While non-Cp Cu was similar in AD and WD, mean value of Cu:Cp distinguished AD from WD (P < .016), with the Cu:Cp Cohen d value of 0.8 (indicating a large biological effect) in AD vs healthy controls, which further increases in WD to 2.2. Concentrations of 24-hour urine copper in 35 healthy controls were compared with those at baseline of an AD patient group analyzed in a previous study on the effect of D-pen on the disease progression. AD patients at baseline had higher concentrations of 24-hour urine copper (P < .001; AD 12.05 µg/day [median, interquartile range 7.85–22.50]) than healthy controls (4.82 µg/day (median, interquartile range: 3.31–7.43]). Values of 24-hour urine copper higher than the cutoff of 200 µg/day were found in 87% of WD and 78% of AD. The direct comparison of AD and WD patients shows a copper failure affecting both diseases, but at a different degree, with the Cu:Cp signifying a more severe grade in WD.

Stroke patients’ clinical course can be largely variable despite similar lesions and clinical states at symptoms’ onset. We sought to identify new serum biomarkers for poststroke functional recovery. We assessed iron, copper, free copper, total antioxidant status (TAS), peroxides, ceruloplasmin (Cp) concentration and activity, ferritin, transferrin (Tf), Cp to Tf ratio (Cp:Tf), and Cp to copper ratio (Cp:Cu) in acute phase (2–10 days from symptoms onset [t0]) in 30 patients affected by stroke in the middle cerebral artery of a single hemisphere. Clinical severity at t0 and at t1 (stabilized phase, typically six months) was evaluated through the National Institute of Health Stroke Scale (NIH) score. Effective recovery (ER) was evaluated as the clinical condition reached in t1 with respect to the condition at symptom onset (NIH1-NIH0/NIH0). In the acute phase (t0), clinical severity correlated with lower levels of TAS and iron, and with increased peroxide concentrations. At t1, clinical severity correlated with increased peroxide and ceruloplasmin concentrations. A worse ER correlated with higher levels of copper, free copper, Cp activity, and values of Cu:Cp. Analysis of oxidative stress and metal profiling of serum could be a useful tool in the search for early biomarkers of ischemic stroke recovery.