Amniotic fluid and bone marrow derived mesenchymal stem cells can be converted to smooth muscle cells in the cryo-injured rat bladder and prevent compensatory hypertrophy of surviving smooth muscle cells

PURPOSE: Wound healing of the cryo-injured bladder can bring about organ remodeling because of incomplete reconstitution of depleted smooth muscle cells. Stem cell transplantation could be beneficial to improve smooth muscle cell regeneration and/or modulate the remodeling process. The repair of bladder injury using adult-type stem cells would be useful for adult urological patients but unsuited for neonatal patients, in whom major benefits are likely to derive from fetal-type stem cells. MATERIALS AND METHODS: The smooth muscle cell differentiation potential of fetal-type vs adult-type stem cells was evaluated by injecting green fluorescent protein labeled mesenchymal stem cells from rat amniotic fluid or bone marrow, respectively, in cryo-injured rat bladder walls. RESULTS: At 30 days after transplantation only a few fetal-type or adult-type mesenchymal stem cells gave rise to enteric or vascular smooth muscle cells, whereas most mesenchymal stem cells appeared incapable of specific differentiation. In vitro co-culture experiments of smooth muscle cells with fetal-type or adult-type mesenchymal stem cells selectively labeled with distinct fluorochromes showed the presence of hybrid cells, suggesting that some mesenchymal stem cells can undergo cell fusion. Surprisingly the major effect of rat bone marrow or amniotic fluid mesenchymal stem cell transplantation seemed to be preventing cryo-injury induced hypertrophy of surviving smooth muscle cells. CONCLUSIONS: In this model stem cell transplantation has a limited effect on smooth muscle cell regeneration. Instead it can regulate post-injury bladder remodeling, possibly via a paracrine mechanism.     

Strengthening acceptance for xenotransplantation: the case of attraction effect

Abstract: Background: Despite being still at the experimental level, xenotransplantation may become an effective strategy to overcome the scarcity of human organs. However, at the present time there is considerable resistance to this kind of biomedical technology. The aim of the present study was to identify novel strategies to reduce patients’ negative affective reactions towards xenotransplantation helping them to understand the advantages of xenotransplantation in a more analytical fashion and increase their acceptance for this approach. Methods: The study was conducted in a group of patients with liver cirrhosis waiting for liver transplantation. They were presented with hypothetical scenarios and asked to choose among either two or three alternative types of donor defined by their species (e.g., livers from humans vs. other species) and availability (low for human donors and high for livers from non‐human species). Results: Patients were unwilling to accept xenotransplantation if they were presented with livers from humans (chosen by 97.5% of participants) vs. livers from genetically modified pigs (2.5%). On the other hand, a different group of patients was significantly more willing to accept xenotransplantation if they were presented with three different types of donors: respectively, human beings (74.4%), genetically modified pigs (25.6%) and genetically modified dogs. In addition, human livers were judged significantly more attractive than genetically modified livers from pigs, monkeys, dogs, or sheep and pig livers were rated as significantly more attractive than livers from monkeys, dogs, or sheep (for all comparisons P < 0.01). Conclusions: These results demonstrate that paradigms from other fields, like decision‐making, might help to communicate more effectively the potential of xenotransplantation, modulating patients’ affective reactions and allowing them to understand the potential strengths of this biomedical technology.     

Cobalt protoporpyhrin reduces caspase-3,-7 enzyme activity in neonatal porcine islets, but does not inhibit cell death induced by TNF-alpha

Apoptotic phenomena observed in vitro following isolation and following transplantation contribute significantly to islet graft loss. Strategies to reduce apoptosis of islet tissue prior to and posttransplantation may improve graft survival and function and reduce the amount of tissue necessary to achieve insulin independence. The expression of cytoprotective proteins is one such strategy that may prolong islet survival. In this light, heme-oxygenase 1 (HO-1) upregulation has been studied in both allo- and xenotransplantation models. In this study, the effect of HO-1 on apoptosis in neonatal porcine islet-like cell clusters (NPICC) was assessed. In in vitro assessments of NPICC apoptosis, NPICC showed a high sensitivity to apoptotic stimulation using a combination of TNF-alpha and cycloheximide. Stimulation with TNF-alpha alone was sufficient to induce reproducible apoptotic responses as demonstrated by caspase-3,-7 activation and subdiploid DNA analysis. Dose-dependent, high-level HO-1 protein expression was achieved following culture of NPICC in medium containing either cobalt protoporphyrin (CoPP) or cobalt mesoporphyrin (CoMP). CoPP treatment resulted in the reduction of caspase-3,-7 enzyme activity following TNF-alpha stimulation. However, such an effect was not associated with a reduction in the levels of cell death. Indeed, the inhibition of caspase enzyme activity resulted in decreased PARP-1 cleavage, which may lead to heightened levels of necrosis in treated NPICC cultures, possibly explaining the observed commitment of NPICC to the death pathway.     

Dynamic contrast enhanced magnetic resonance imaging of the terminal ileum: differentiation of activity of Crohn’s disease

AIM: To prospectively investigate a new high resolution MRI technique for dynamic evaluation of the enhancement kinetics of bowel parietal layers and to correlate it with CDAI, CRP, endoscopic activity and histologic features. METHODS: About 16 consecutive patients with proven diagnosis of CD underwent ileocolonoscopy with biopsy and serial bowel dynamic contrasted-MRI (D-CE-MRI) evaluated in blind fashion. Quantitative analysis of bowel wall enhancement kinetics was performed basing on signal to noise ratio (SNR) of inner parietal layers (Mucosa-Submucosa, M-SM) and outer parietal layers (Muscular-Serosa, Ms-S). Disease activity was defined by CDAI > 150, serum CRP > 5 mg/dL and histologic results. RESULTS: About 9 patients showed a layered enhancement of bowel wall (8 active, 1 inactive), whereas inactive (7 cases) group presented a homogeneous pattern. In active patients we found a significant difference in parietal layered enhancement curves (M-SM vs. Ms-S, P < 0.03) not observed in inactive disease and controls (intra-group analysis). M-SM and Ms-S enhanced curves in clinically active patients were significantly different respect to those of patients with inactive CD (P < 0.001) (inter-group analysis). Parietal D-CE-MRI pattern well correlated with histologic features (r = 0.8; P < 0.001, Spearman test). CONCLUSIONS: D-CE-MRI can be a useful tool for clinical follow-up and in the treatment strategies in CD patients.     

Isolated knee monoarthritis heralding resectable non-small-cell lung cancer. A paraneoplastic syndrome not previously described

Objective

To describe isolated knee monoarthritis as a paraneoplastic syndrome heralding non‐small cell lung cancer (NSCLC), and to discuss its clinical characteristics.

Methods

Clinical records of all consecutive, new outpatients with isolated knee monoarthritis observed from January 2000 to December 2005 were reviewed. A systematic review of Medline and Cochrane Library databases was performed to identify English‐language articles related to rheumatological paraneoplastic syndromes associated with NSCLC.

Results

Over 6 years, 6654 new outpatients with different rheumatic disorders were observed. Of these, 296 (4.4%) presented with isolated monoarthritis of the knee. In five out of 296 patients (1.7%) this feature represented the initial manifestation of NSCLC. All five patients were middle‐aged men, with a long history of heavy cigarette smoking, who had a non‐erosive, isolated knee monoarthritis, with mild articular fluid collection of non‐inflammatory type. NSCLC was resectable in all patients, and knee monoarthritis remitted with no relapse confirming its paraneoplastic nature. All five patients are in good condition after a median follow up of 41 months. The literature review revealed that paraneoplastic knee monoarthritis has not previously been reported.

Conclusion

Knee monoarthritis may in some cases represent a paraneoplastic syndrome heralding NSCLC at an early stage.Small‐cell lung cancer (SCLC) is associated with various types of paraneoplastic syndromes in approximately 10–20% of cases.^1,2 Paraneoplastic syndromes associated with non‐small‐cell lung cancer (NSCLC) are less frequent and variable, but of particular interest. Given its resistance to chemotherapy, the prognosis of NSCLC is strictly dependent on the possibility of surgical removal, its early detection is thus of great importance.³Hypertrophic osteoarthropathy (HOA), observed in approximately 5% of patients, is the most frequent paraneoplastic syndrome associated with NSCLC.^1,2To the best of our knowledge, isolated knee monoarthritis not associated with HOA, as the presenting feature of NSCLC, has not previously been reported.A review of the clinical records of all outpatients presenting with isolated knee monoarthritis observed over a 6‐year period revealed that this manifestation heralded NSCLC in five patients.The aim of this study was to describe the clinical characteristics of these patients, and to discuss the differential diagnosis.     

Virtual screening for novel openers of pancreatic K(ATP) channels

Ligand-based virtual screening approaches were applied to search for new chemotype KCOs activating Kir6.2/SUR1 KATP channels. A total of 65 208 commercially available compounds, extracted from the ZINC archive, served as database for screening. In a first step, pharmacokinetic filtering via VolSurf reduced the initial database to 1913 compounds. Afterward, six molecules were selected as templates for similarity searches: similarity scores, obtained toward these templates, were calculated with the GRIND, FLAP, and TOPP approaches, which differently encode structural information into potential pharmacophores. In this way, we obtained 32 hit candidates, 16 via GRIND and eight each via FLAP and TOPP. For biological testing of the hit candidates, their effects on membrane potentials in HEK 293 cells expressing Kir6.2/SUR1 were studied. GRIND, FLAP, and TOPP all yielded hits, but no method top-ranked all the actives. Thus, parallel application of different approaches probably improves hit detection.     

Establishing an Italian general practitioner brief intervention pilot project for problem drinkers

This national pilot project commenced in 2004. It was promoted by the Istituto Superiore di Sanità and financed by the Italian Ministry of Health. The Italian Society of General Practitioners supported implementation of the study in general medical practitioner settings and coordination was undertaken by the Alcohol Centre of the University Hospital of Florence. The objectives of the study are to create a brief intervention package, suitable for use within the Italian primary health care system for screening and treating "hazardous drinkers"; to survey alcohol consumption in a sample of the population aged 18 or over, who the general practitioners assessed as suitable for brief intervention using the project's instruments; to conduct an experimental trial of brief intervention procedures, with "hazardous drinkers" randomly allocated to intervention or control conditions; to assess the effectiveness of brief intervention as a primary preventive measure. The early stage of this study concentrated on creating a strong research partnership to foster the involvement of general medical practitioners on a national scale. Subsequently, resources were devoted to the creation of the brief intervention package, including support material and the training of the general medical practitioners in its implementation.     

Spontaneous cerebrospinal fluid rhinorrhea as the presenting symptom of sellar pathologies: three demonstrative cases

Spontaneous rhinorrhea due to sellar pathologies is a rare and insidious disease that may represent a diagnostic challenge. Since the precipitating cause is not apparent in most patients, delayed diagnosis and/or improper treatments are not uncommon and may be detrimental for the patients. The precise mechanisms of such rhinorrhea are still incompletely understood. Proposed etiological factors include constant cerebrospinal fluid pulsations against the anterior skull base trasmitted by primitive or acquired arachnoid invaginations in combination with either elevated ICP or congenital/pathologic erosions of sellar and parasellar bone structures. Advances in the diagnosis and surgical techniques have recently modified the decision-making approach to this pathology. The present paper reports three unusual cases of sellar pathologies revealed by rhinorrhea as the first symptom discussing controversial issues on pathogenesis, complications and current management of this kind of fistula.     

An emergency clinical pathway for stroke patients – results of a cluster randomised trial (isrctn41456865)

Background  

Emergency Clinical Pathways (ECP) for stroke have never been tested in randomized controlled trials (RCTs).