Costantino Santacroce

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.

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.

As previously described by several authors, dental pulp stem cells (DPSCs), when adequately stimulated, may acquire a neuronal-like phenotype acting as a favorable source of stem cells in the generation of nerves. Besides, it is known that hypoxia conditioning is capable of stimulating cell differentiation as well as survival and self-renewal, and that multiple growth factors, including Epidermal Growth factor (EGF) and basic fibroblast growth factor (bFGF), are often involved in the induction of the neuronal differentiation of progenitor cells. In this work, we investigated the role of hypoxia in the commitment of DPSCs into a neuronal phenotype. These cells were conditioned with hypoxia (O2 1%) for 5 and 16 days; subsequently, we analyzed the proliferation rate and morphology, and tested the cells for neural and stem markers. Moreover, we verified the possible autocrine/paracrine role of DPSCs in the induction of neural differentiation by comparing the secretome profile of the hypoxic and normoxic conditioned media (CM). Our results showed that the hypoxia-mediated DPSC differentiation was time dependent. Moreover, conditioned media (CM derived from DPSCs stimulated by hypoxia were able, in turn, to induce the neural differentiation of SH-SY5Y neuroblastoma cells and undifferentiated DPSCs. In conclusion, under the herein-mentioned conditions, hypoxia seems to favor the differentiation of DPSCs into neuron-like cells. In this way, we confirm the potential clinical utility of differentiated neuronal DPSCs, and we also suggest the even greater potential of CM-derived-hypoxic DPSCs that could more readily be used in regenerative therapies.

Human Dental Pulp Stem Cells (hDPSCs) represent a type of adult mesenchymal stem cells that have the ability to differentiate in vitro in several lineages such as odontoblasts, osteoblasts, chondrocytes, adipocytes and neurons. In the current work, we used hDPSCs as the experimental model to study the role of recombinant prion protein 23⁻231 (recPrPC) in the neuronal differentiation process, and in the signal pathway activation of ERK 1/2 and Akt. We demonstrated that recPrPC was able to activate an intracellular signal pathway mediated by extracellular-signal-regulated kinase 1 and 2 (ERK 1/2) and protein kinase B (Akt). Moreover, in order to understand whether endogenous prion protein (PrPC) was necessary to mediate the signaling induced by recPrPC, we silenced PrPC, demonstrating that the presence of endogenous PrPC was essential for ERK 1/2 and Akt phosphorylation. Since endogenous PrPC is a well-known lipid rafts component, we evaluated the role of these structures in the signal pathway induced by recPrPC. Our results suggest that lipid rafts integrity play a key role in recPrPC activity. In fact, lipid rafts inhibitors, such as fumonisin B1 and MβCD, significantly prevented ERK 1/2 and Akt phosphorylation induced by recPrPC. In addition, we investigated the capacity of recPrPC to induce hDPSCs neuronal differentiation process after long-term stimulation through the evaluation of typical neuronal markers expression such as B3-Tubulin, neurofilament-H (NFH) and growth associated protein 43 (GAP43). Accordingly, when we silenced endogenous PrPC, we observed the inhibition of neuronal differentiation induced by recPrPC. The combined data suggest that recPrPC plays a key role in the neuronal differentiation process and in the activation of specific intracellular signal pathways in hDPSCs.

Human dental pulp-derived stem cells (hDPSCs) are characterized by a typical fibroblast-like morphology. They express specific markers for mesenchymal stem cells and are capable of differentiation into osteoblasts, adipoblasts and neurons in vitro. Previous studies showed that gangliosides are involved in the induction of early neuronal differentiation of hDPSCs. This study was undertaken to investigate the role of lipid rafts in this process. Lipid rafts are signaling microdomains enriched in glycosphingolipids, cholesterol, tyrosine kinase receptors, mono- or heterotrimeric G proteins and GPI-anchored proteins. We preliminary showed that established cells expressed multipotent mesenchymal stromal-specific surface antigens. Then, we analyzed the distribution of lipid rafts, revealing plasma membrane microdomains with GM2 and EGF-R enrichment. Following stimulation with EGF/bFGF, neuronal differentiation was observed. To analyze the functional role of lipid rafts in EGF/bFGF-induced hDPSCs differentiation, cells were preincubated with lipid raft affecting agents, i.e. [D]-PDMP or methyl-β-cyclodextrin. These compounds significantly prevented neuronal-specific antigen expression, as well as Akt and ERK 1/2 phosphorylation, induced by EGF/bFGF, indicating that lipid raft integrity is essential for EGF/bFGF-induced hDPSCs differentiation. These results suggest that lipid rafts may represent specific chambers, where multimolecular signaling complexes, including lipids (gangliosides, cholesterol) and proteins (EGF-R), play a role in hDPSCs differentiation.

Cellular prion protein (PrPC) is expressed in a wide variety of stem cells in which regulates their self-renewal as well as differentiation potential. In this study we investigated the presence of PrPC in human dental pulp-derived stem cells (hDPSCs) and its role in neuronal differentiation process. We show that hDPSCs expresses early PrPC at low concentration and its expression increases after two weeks of treatment with EGF/bFGF. Then, we analyzed the association of PrPC with gangliosides and EGF receptor (EGF-R) during neuronal differentiation process. PrPC associates constitutively with GM2 in control hDPSCs and with GD3 only after neuronal differentiation. Otherwise, EGF-R associates weakly in control hDPSCs and more markedly after neuronal differentiation. To analyze the functional role of PrPC in the signal pathway mediated by EGF/EGF-R, a siRNA PrP was applied to ablate PrPC and its function. The treatment with siRNA PrP significantly prevented Akt and ERK1/2 phosphorylation induced by EGF. Moreover, siRNA PrP treatment significantly prevented neuronal-specific antigens expression induced by EGF/bFGF, indicating that cellular prion protein is essential for EGF/bFGF-induced hDPSCs differentiation. These results suggest that PrPC interact with EGF-R within lipid rafts, playing a role in the multimolecular signaling complexes involved in hDPSCs neuronal differentiation.

Gangliosides (GGs) are a glycolipid class present on Mesenchymal Stem Cells (MSCs) surfaces with a critical appearance role in stem cell differentiation, even though their mechanistic role in signaling and differentiation remains largely unknown. This review aims to carry out a critical analysis of the predictive role of gangliosides as specific markers of the cellular state of undifferentiated and differentiated MSCs, towards the osteogenic, chondrogenic, neurogenic, and adipogenic lineage. For this reason, we analyzed the role of GGs during multilineage differentiation processes of several types of MSCs such as Umbilical Cord-derived MSCs (UC-MSCs), Bone Marrow-derived MSCs (BM-MSCs), Dental Pulp derived MSCs (DPSCs), and Adipose derived MSCs (ADSCs). Moreover, we examined the possible role of GGs as specific cell surface markers to identify or isolate specific stem cell isotypes and their potential use as additional markers for quality control of cell-based therapies.

Bioethical issues related to the manipulation of embryonic stem cells have hindered advances in the field of medical research. For this reason, it is very important to obtain adult stem cells from different tissues such as adipose, umbilical cord, bone marrow and blood. Among the possible sources, dental pulp is particularly interesting because it is easy to obtain in respect of bioethical considerations. Indeed, human Dental Pulp Stem Cells (hDPSCs) are a type of adult stem cells able to differentiate in neuronal-like cells and can be obtained from the third molar of healthy patients (13-19 ages). In particular, the dental pulp was removed with an excavator, cut into small slices, treated with collagenase IV and cultured in a flask. To induce the neuronal differentiation, hDPSCs were stimulated with EGF/bFGF for 2 weeks. Previously, we have demonstrated that during the differentiation process the content of cellular prion Protein (PrPC) in hDPSCs increased. The cytofluorimetric analysis showed an early expression of PrPC that increased after neuronal differentiation process. Ablation of PrPC by siRNA PrP prevented neuronal differentiation induced by EGF/bFGF. In this paper, we illustrate that as we enhanced the isolation, separation and in vitro cultivation methods of hDPSCs with several easy procedures, more efficient cell clones were obtained and large-scale expansion of the mesenchymal stem cells (MSCs) was observed. We also show how the hDPSCs, obtained with methods detailed in the protocol, are an excellent experimental model to study the neuronal differentiation process of MSCs and subsequent cellular and molecular processes.

The prion protein (PrP) is an enigmatic molecule with a pleiotropic effect on different cell types; it is localized stably in lipid raft microdomains and it is able to recruit downstream signal transduction pathways by its interaction with various biochemical partners. Since its discovery, this lipid raft component has been involved in several functions, although most of the publications focused on the pathological role of the protein. Recent studies report a key role of cellular prion protein (PrPC) in physiological processes, including cellular differentiation. Indeed, the PrPC, whose expression is modulated according to the cell differentiation degree, appears to be part of the multimolecular signaling pathways of the neuronal differentiation process. In this review, we aim to summarize the main findings that report the link between PrPC and stem cells.

Abstract Mesenchymal stem cells (MSCs) are well known for their beneficial effects, differentiation capacity and regenerative potential. Dental-derived MSCs (DSCs) are more easily accessible and have a non-invasive isolation method rather than MSCs isolated from other sources (umbilical cord, bone marrow, and adipose tissue). In addition, DSCs appear to have a relevant neuro-regenerative potential due to their neural crest origin. However, it is now known that the beneficial effects of MSCs depend, at least in part, on their secretome, referring to all the bioactive molecules (neurotrophic factors) released in the conditioned medium (CM) or in the extracellular vesicles (EVs) in particular exosomes (Exos). In this review, we described the similarities and differences between various DSCs. Our focus was on the secretome of DSCs and their applications in cell therapy for neurological disorders. For neuro-regenerative purposes, the secretome of different DSCs has been tested. Among these, the secretome of dental pulp stem cells and stem cells from human exfoliated deciduous teeth have been the most widely studied. Both CM and Exos obtained from DSCs have been shown to promote neurite outgrowth and neuroprotective effects as well as their combination with scaffold materials (to improve their functional integration in the tissue). For these reasons, the secretome obtained from DSCs in combination with scaffold materials may represent a promising tissue engineering approach for neuroprotective and neuro-regenerative treatments. Graphical Abstract

The Ku protein is a tightly associated heterodimer, comprised of 70-kilodalton (kD) and 86-kD subunits, that forms the DNA-dependent protein kinase (DNA-PK) complex together with the 470-kD DNA-PKcs catalytic subunit, and is involved mainly in DNA double-strand breaks (DSBs) repair. The objective of the current study was to investigate the expression and DNA-binding activity of the Ku protein in fresh tissues from patients with bladder carcinoma and to compare it with that in nontumor tissues obtained from the same organ. Moreover, the DNA-binding activity of Ku was assessed after exposure of the tumor cells to 1 or 2 grays (Gy) of X-rays. Furthermore, the level of phosphorylated Ku was analyzed in both the nuclear and cytoplasmic compartment of normal tissue after exposure to 2 Gy of X-rays.The expression and DNA-binding activity of Ku protein were assessed in tumor samples from patients who all were diagnosed with transitional cell carcinoma (TCC) of the bladder using Western blot analysis and the electrophoretic mobility shift assay, respectively.Enhanced Ku activity and expression were found in tumor tissue compared with normal tissue for each patient. Moreover, variations in Ku activity were found in a dose-dependent manner after the tumor cells were exposed to 1 or 2 Gy of X-rays. A decrease in phosphorylated Ku in the cytoplasm and a parallel increase in the nucleus of normal tissue cells were observed after exposure to X-rays.The results of the current study suggest a possible role of Ku in regulating the DNA-PK activity of DSBs repair in bladder tumors.

A 69-year-old man was admitted with a complaint of left irreducible inguinal mass. On surgical exploration no evidence of hernia was found and the inguinal floor was overwhelmed by a large lobulated mass, arising from the properitoneal fat, that involved the spermatic cord. The mass was partially removed, sparing the elements of cord. The transversalis fascia was repaired by direct suture and a polypropylene mesh was located above. The histopathological diagnosis was well differentiated-type liposarcoma with myxoid features. The liposarcoma is a malignant tumour of the adipose tissue that arises from the primitive mesenchymal cells. These neoplasms have been usually found in the soft tissues of limbs, trunk, mediastinum, retroperitoneum and occasionally in the spermatic cord. The clinical aspect is frequently a complaint of scrotal or inguinal painless mass, mimicking to an inguinal hernia and the diagnosis of tumor is performed mainly during surgery, as in our patient. In the case of a firm not reducible painless inguinal mass without signs and symptoms of bowel obstruction, an abdominal tumor with inguinal or scrotal extension should be suspected and preoperatively excluded. The US and CT scan may be helpful to plane a correct therapeutic strategy before intervention.

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.

Panniculitides represent a heterogeneous group of inflammatory diseases involving subcutaneous fat. Subcutaneous fat is normally organized into adipose cells, adipocytes, and septa of connective tissue. The inflammation involving such tissues can be more represented in septa (septal panniculitis) or in lobules (lobular panniculitis) or be equally distributed in both (mixed panniculitis). A bioptical study is necessary in order to discern between different forms. Vascular involvement is also different in such diseases, as it can interest arteries, or veins, or both. Different grades of fat necrosis can also be observed, such as adipocytes without nuclei, lipophagic necrosis, liquefactive fat necrosis, microcystic fat necrosis, ischaemic fat necrosis. Panniculitis can be idiopathic or secondary to other diseases such as systemic sclerosis, rheumatoid arthritis, systemic erithematous lupus and many others. Therapies usually vary on the single patient but the general orientation leads to the use of immunosuppressive drugs such as thalidomide, corticosteroids, cyclosporin-A, hydroxychloroquine and cyclophosphamide. We report a case of a 19-year-old female affected by primary mixed panniculitis, associated with fever and deep asthenia, that resolved in a few weeks and was maintained with oral cyclosporin-A.

Cell outer membranes contain glycosphingolipids and protein receptors, which are integrated into glycoprotein domains, known as lipid rafts, which are involved in a variety of cellular processes, including receptor-mediated signal transduction and cellular differentiation process. In this study, we analyzed the lipidic composition of human Dental Pulp-Derived Stem Cells (DPSCs), and the role of lipid rafts during the multilineage differentiation process. The relative quantification of lipid metabolites in the organic fraction of DPSCs, performed by Nuclear Magnetic Resonance (NMR) spectroscopy, showed that mono-unsaturated fatty acids (MUFAs) were the most representative species in the total pool of acyl chains, compared to polyunsatured fatty acids (PUFAs). In addition, the stimulation of DPSCs with different culture media induces a multilineage differentiation process, determining changes in the gangliosides pattern. To understand the functional role of lipid rafts during multilineage differentiation, DPSCs were pretreated with a typical lipid raft affecting agent (MβCD). Subsequently, DPSCs were inducted to differentiate into osteoblast, chondroblast and adipoblast cells with specific media. We observed that raft-affecting agent MβCD prevented AKT activation and the expression of lineage-specific mRNA such as OSX, PPARγ2, and SOX9 during multilineage differentiation. Moreover, this compound significantly prevented the tri-lineage differentiation induced by specific stimuli, indicating that lipid raft integrity is essential for DPSCs differentiation. These results suggest that lipid rafts alteration may affect the signaling pathway activated, preventing multilineage differentiation.

To evaluate CVVH (Continuous Veno-Venous Hemofiltration) as acute renal replacement treatment in postoperative care of liver transplantation.Retrospective study.Intensive Care Unit, year 1995.86 OLT performed in 1995, 11 of them underwent acute renal replacement treatment. In the same period, in the ICU were admitted 237 patients, and 20 underwent acute renal replacement treatment (control group). Evaluation with SOFA (Sepsis-related Organ Failure Assessment) score.CVVH performed heparin free, pump system, polyamide or polysulphone 0.6 mq membrane hemofilter device, blood flow 150-200 ml/min, UF rate 1000-1200 ml/h, clearance 16-20 ml/min.Coagulation monitoring (PT as INR, PTT, fibrinogen, antithrombin III, d-dimer, platelet count) was performed 3 times a day or on variation of the clinical conditions.SOFA score did not differ between the two groups. Mortality was higher in the patients treated with CVVH. CVVH was performed from 16 to 18 hrs/day for 9.90 +/- 2.33 days. Three patients developed clinical bleeding before CVVH, 3 during CVVH but 1 of them underwent repeated surgical procedures.We cannot demonstrate that CVVH doesn't affect bleeding, but we can say that, for the complexity of the post OLT patients, CVVH can be the treatment of choice in case of renal replacement treatment.

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.

475 patients with carcinoma at different sites (141 colon-rectum; 102 breast; 50 stomach; 48 kidney; 46 head and neck; 41 bladder; 47 other sites) submitted to surgery have been analyzed after histopathological staging and grading, by flow cytometry (monoparametric DNA content analysis) and immunohistochemistry (p53, c-erbB-2, and PCNA expression). In breast cancer patients the presence of receptors for estrogen (ER) and progesterone (PGR) has also been determined. Flow cytometry-derived parameters were DNA ploidy, fraction of cells in S-phase (SPF), and DNA content heterogeneity (multi-clonal stem cell lines with different DNA index and/or more than one subpopulations with different ploidy levels in different samples from the same tumor). Correlations of the results obtained by the different techniques have been attempted by the non-parametric Spearman's rank correlation approach. Significant associations (P < 0.05) were found between the histopathological, immunohistochemical and flow cytometric parameters considered in some anatomical regions, such as stomach (p53 vs DNA content aneuploidy and vs heterogeneity), colon-rectum (TNM vs p53 and vs heterogeneity), bladder (grading vs DNA content aneuploidy and vs heterogeneity). Tumor heterogeneity proved to be dependent on the number of tumor samples taken. The results of this preliminary assessment will subsequently be compared with the data obtained from a currently ongoing follow-up survey.

Meconium staining of the fetus and placenta is associated with increased neonatal mortality and asphyxia. Very often it is unclear whether the discharge of meconium is a cause or an effect of fetal distress. In the available literature there are no large epidemiological studies of pregnancy outcome with meconium-related lesions, even though this could be useful to improve our state of knowledge on this topic.A case of umbilical cord vascular necrosis is described. A severely asphyxiated infant was delivered at 39 weeks' gestation by cesarean section due to alarming results of fetal heart rate monitoring and rupture of membranes with meconium-stained amniotic fluid. There was no meconium aspiration. We report a review of 15 similar cases. In the whole series, a linkage between umbilical cord vascular necrosis and evidence of remote meconium discharge always seems to be detectable. The pathophysiological mechanism is unknown.It is still not clear why only a tiny percentage of cases with meconium-stained amniotic fluid develops umbilical cord lesions and poor pregnancy outcome. Further investigations are needed to explain why some meconium-stained newborns suffer severe neurological and other damage even without meconium aspiration.

Background: The therapeutic ERCP before the VLC in the patients with moderate-severe acute biliary pancreatitis (ABP) is a well recognized practice, instead the necessity of ERCP in the patients with mild acute biliary pancreatitis is not well defined. Aim: The aim of this study was to evaluate the usefulness of the MRCP before the VLC in the patients with mild acute biliary pancreatitis. Methods: In the period 2003-2006, 25 patients were submitted to a MRCP (F/M:15/10) with mild ABP without increase of the cholestasis tests and absence of choledocholithiasis at the abdominal USG. During a follow-up from 15 to 60 days after the VLC, the presence of jaundice or relapse of ABP were evaluated in all patients by means clinical and/or laboratory and/or instrumental examinations. Results: Six patients had diagnosis of choledocholithiasis (stones, biliary sand or biliary sludge) at the MRCP and they were submitted to an ERCP with papillotomy and stones removal; 19 patients with a negative MRCP were submitted to the VLC. All the 25 patients submitted to the MRCP before the VLC did not have jaundice or relapse of the ABP during the follow-up period. Conclusions: The MRCP was an accurate investigation for the preoperatory diagnosis of choledocholithiasis; even if it is not possible to recommend its utilization extensively, it is an important procedure for the patients with diagnosis of mild ABP to select all those to submit to the ERCP.

Aims: The literature does not define unanimously if the VLC must be executed just after the normalization (objective and subjective) of the abdominal symptomatology or after the normalization of the lypasemia, also. The aim of this study was to define the optimal timing of the VLC for the patients with mild acute biliary pancreatitis. Methods: In the period 1997-2006, forty patients with mild acute biliary pancreatitis (Glasgow's criteria) were admitted within our Institution: 29 females, 11 males, mean age 57 years (range 26-78 years); 9 patients with diagnosis of principal biliary duct lithiasis, were submitted to an ERCP with papillotomy and stones or biliary sludge/sand removal; in 31 patients without choledocholithiasis (stones or biliary sand/sludge) revealed by laboratory and instrumental tests were subdivided in two homogeneous groups about the age, sex and clinical, laboratory and instrumental conditions. Sixteen patients (first group) underwent a VLC after 24 hours from the resolution (objective and subjective) of the abdominal symptomology together with the decrease (not normalization) of the lypasemia (4 days in mean, range 2-7 days); 15 patients (second group) underwent a VLC after 24 hours from the normalization of the lypasemia together with the absence of abdominal symptomatology (7 days in mean, range 4-13 days). Results: There were no intraoperative complications or major postoperative morbidity in both groups. The total hospital stay was 5 days in mean for the first group (range 3-10 days) and 9 days for the second group (range 6-15 days). Conclusions: In our opinion, according to our results, the VLC can be safely executed after 24 hours from the objective/subjective resolution of the abdominal symptomatology, without the normalization of the lypasemia, in patients with mild acute biliary pancreatitis, reducing in this way the total hospital staying.

Introduction: There is not uniformity in the literature about the timing of execution of the endoscopic sphincterotomy (ES) and later the videolaparocholecystectomy (VLC) in course of acute biliary pancreatitis (ABP): the aim of the study was to suggest the optimal timing. Patients and Methods: In the period September 1997-November 2004, 67 patients were treated for ABP. Fifteen cases were severe ABP and 52 were mild ABP. In 55 patients an ES was executed within 48-72 hours; in 12 cases the ES was delayed of 10 days. After 8-10 days, 62 patients had a VLC; 5 patients had an open cholecystectomy. Results: The coledochal stone was removed in 32 cases (47.7%); in the last 35 patients (52.3%) the biliary sand or sludge was removed. Immediate results: 1 case (1.5%) of pancreatitis reacutizzation, 6 cases (9.8%) increase of the lypase and amylase, 2 (2.9%) duodenal perforations. The cholecystectomy, laparoscopic and open, did not have relevant complications. Later in time we have registered the develop of 3 postnecrotic pancreatic pseudocysts, treated with surgical therapy. Conclusions: The ES in course of severe and mild ABP has the double goal to clean the principal biliary duct by the stones, and moreover to remove the papillar obstacle because of stenosis, biliary sand or sludge. In our experience, the golden therapeutic timing foresees the ES within 48-72 hours from the beginning of the symptomatology and the VLC within 8-10 days: this time is necessary to establish the absence the progression of the acute pancreatitis.

Bioethical issues related to the manipulation of embryonic stem cells have hindered advances in the field of medical research. For this reason, it is very important to obtain adult stem cells from different tissues such as adipose, umbilical cord, bone marrow and blood. Among the possible sources, dental pulp is particularly interesting because it is easy to obtain in respect of bioethical considerations. Indeed, human Dental Pulp Stem Cells (hDPSCs) are a type of adult stem cells able to differentiate in neuronal-like cells and can be obtained from the third molar of healthy patients (13-19 ages). In particular, the dental pulp was removed with an excavator, cut into small slices, treated with collagenase IV and cultured in a flask. To induce the neuronal differentiation, hDPSCs were stimulated with EGF/bFGF for 2 weeks. Previously, we have demonstrated that during the differentiation process the content of cellular prion Protein (PrPC) in hDPSCs increased. The cytofluorimetric analysis showed an early expression of PrPC that increased after neuronal differentiation process. Ablation of PrPC by siRNA PrP prevented neuronal differentiation induced by EGF/bFGF. In this paper, we illustrate that as we enhanced the isolation, separation and in vitro cultivation methods of hDPSCs with several easy procedures, more efficient cell clones were obtained and large-scale expansion of the mesenchymal stem cells (MSCs) was observed. We also show how the hDPSCs, obtained with methods detailed in the protocol, are an excellent experimental model to study the neuronal differentiation process of MSCs and subsequent cellular and molecular processes.

An increased risk of adenocarcinoma of the oesophagus has been demonstrated in patients with long segments of Barrett's mucosa. The risk of cancer associated with short segments of metaplasia of the oesophagogastric junction is not known.We report a case of early adenocarcinoma of the oesophagus arising on short tongues of Barrett's mucosa associated with an oesophageal cyst. The patient was a 68-year-old man with no previous clinical history of gastro-oesophageal reflux disease. The fortuitous discovery of an oesophageal cyst lead to the diagnosis of short tongues of Barrett's mucosa with high-grade dysplasia. On pathological examination of the resected specimen, an early adenocarcinoma had developed in Barrett's mucosa, localized just above the oesophageal cyst.As oesophageal cysts can cause symptoms suggestive of reflux, we hypothesize that this association may not be fortuitous.

Aim: Studies that associate the hemoconcentration with the development of necrotizing acute pancreatitis are present in literature. The aim of this study is to evaluate if the hemoconcentration is an early marker of necrotizing pancreatitis. Patients and Methods: The study was executed on patients admitted in the Division of General Surgery of Foggia University with diagnosis of acute pancreatitis in the period from January 1998 to June 2005. The prognostic Ranson's criteria were applied in all patients. Among the 60 patients admitted with diagnosis of acute pancreatitis (biliary pancreatitis in almost all patients), 24 were submitted to a CT-scan within 36-72 hours. Seven of the 24 patients had a necrotizing pancreatitis (Balthazar's score). The hematocrit (Hct) was retrospectively evaluated and associated with the CT-scan morphological data, as an early marker of pancreatitis severity. Results: The regression analysis showed an association between pancreatic necrosis, by means of CT evaluation, and the hemoconcentration. Hematocrit more than 43% in the males and more than 39% in the females and/or a reduction of the Hct within the first 24 hours from the admission, were markers of severity and pancreatic necrosis (and organ failure). In 6 of the 7 patients with necrotizing pancreatitis there was critical value of Hct and only in 4 of the 17 patients with edematous pancreatitis there was a high value of Hct, showing the statistical significance of the proposed criteria (p < 0.01). The negative predictive value of the hemoconcentration was 94.7% for the evolution in pancreatic necrosis. Conclusions: The prognostic value of the hemoconcentration is comparable with the score of Ranson (48 hours of observation). So, it is an early and simple marker of the necrotizing evolution of the acute pancreatitis, because of its high negative predictive value: the patients with acute pancreatitis without hemoconcentration will rarely develop a necrotizing pancreatitis.

Background: Dental erosions are assessed complications of GORD and gingival aphtosis may be concurrent with CD.(#) Aim of this study was to investigate the prevalence of GORD and CD in patients with oro-dental lesions. Materials and Methods: Between september 2003 and 2004, 120 patients underwent dental examination for the first time by a senior dental physician and if positive were referred to our Gastroenterology Unit. Presence and localization of dental erosions was assessed by means of Eccles and Jenkins scale (A group). On the other hand were evaluated aphtoid lesions with a story of two events per month or more (B group). The A pts only if 40 years old or more, were directed to perform trans-nasal gastroscopy (TN-EGDS) and if negative 24 hours pH-metry; urea breath test (UBT) was tested. The B pts were requested to measure out anti-gliadin, anti-endomisium, anti-tissutal transglutaminase antibodies and if positive, to perform TN-EGDS with duodenal bioptic study. Results: In 41/120 patients (29.2%, M/F = 16/25, range 9-70 yy) oro-dental lesions were detected. 27/41 pts (M/F:10/17) underwent gastroenterological examination, 22(M/F:7/15) in A group and 5 in B group (M/F:3/2). In the A group 5 symptomatic pts refused EGDS and 1female was rejected because anorexic. 14/16 pts (87.5%, M/F:6/8 range 29-73 yy ma:43,43) had esophagitis or pathologic reflux at pH-metry; UBT was positive in only 1 pt. In the B group we detected 2 CD both UBT negative: 1M 25 and 1F 24 years old. Conclusions: Our preliminary experience suggest: 1) an high agreement between dental erosions and GORD. 2) gingival aphtosis is an accurate marker of CD. In the light of these results the role of the dentist could support the gastroenterologist's and the involvement of several specialists will make screening possible on a large scale in future. (#) Jarvinen V et Al. Caries Res. 1992;26(5):391-6.

It is known that in the course of osteoarthritis (OA), articular cartilage develops biochemical and structural changes. In the last years, serum and urinary markers of both the synthesis and destruction of cartilage have been dosed, above all in order to carry out an early diagnosis of OA. Among them, the urinary excretion of pyridinoline seems to correlate with the entity of the degradation of cartilage. The aim of the present study is to evaluate the above mentioned markers in OA patients compared to control subjects. Moreover, the possible influence on cartilage of two different non steroidal antiinflammatory drugs (NSAIDs), in particular Nabumetone and Piroxicam, has been verified. The study shows that the urinary excretion of pyridinoline is able to express the severity of OA. At last, the study shows that the tested drugs do not interfere with the metabolism of cartilage.