Cloning of the rat ADAMTS‐1 gene and its down regulation in endothelial cells in cirrhotic rats

Abstract: Aims: This study was undertaken in order to identify genes which are regulated during the process of liver fibrosis. Methods: The differential display method and RNA from rat endothelial cells before and after induction of cirrhosis were used. Results: A 496bp fragment, which was down regulated in liver endothelial cells from a cirrhotic animal, was cloned. The cloned fragment showed a 95% homology with the newly cloned mouse ADAMTS‐1 gene (a disintegrin and metalloproteinase with thrombospondin motifs), which is implicated in inflammation. The fragment was found to span the 3′ of exon 6, the whole exon 7 and the 5′ of exon 8. Sequencing of the entire coding region of the rat gene showed a 94% homology at the nucleic acid level and 96% homology at the amino acid level. The sequences responsible for the function of the protein were conserved. Northern blot analysis, using the cloned fragment as a probe, confirmed the finding that the gene was down‐regulated in endothelial cells derived from livers of cirrhotic animals. In situ PCR analysis localised the ADAMTS‐1 gene in the liver endothelial cells from normal animals. Conclusions: Regulation of the expression of genes which belong to the metalloproteinase family in liver endothelial cells might be important in the development of liver cirrhosis.     

The role of upper endoscopy in identifying oesophageal involvement in patients with oral pemphigus vulgaris

BACKGROUND AND AIMS: The involvement of oesophagus in pemphigus vulgaris is still debated. The aims of this study were to evaluate the prevalence of oesophageal involvement and the gastro-duodenal mucosa appearance before and after high-dose corticosteroid therapy in a group of patients with oral pemphigus vulgaris. METHODS: We prospectively studied 28 consecutive patients with oral pemphigus by oesophageal symptom standardised questionnaire, upper gastro-intestinal endoscopy, exfoliative cytology and histological biopsy. After clinical remission, all patients underwent new endoscopy. RESULTS: The prevalence of oesophageal symptoms was 57.1%. Endoscopic examination revealed oesophageal involvement with different degrees of severity in 67.8% of patients. After corticosteroid therapy, endoscopy showed normal oesophageal-gastro-duodenal mucosa. No examination-related exacerbations of the oesophageal lesions were seen. CONCLUSIONS: The upper gastro-intestinal endoscopic examination, in oral pemphigus vulgaris patients with oesophageal symptoms, is safe in skilled hands technique and a useful diagnostic tool prior to starting therapy.     

Colistin Methanesulfonate and Colistin Pharmacokinetics in Critically Ill Patients Receiving Continuous Venovenous Hemodiafiltration

This report describes the pharmacokinetics of colistin methanesulfonate (CMS) and colistin in five intensive care unit patients receiving continuous venovenous hemodiafiltration. For CMS, the mean maximum concentration of drug in plasma (C_max) after the fourth dose was 6.92 mg/liter and total clearance (CL) 8.23 liters/h. For colistin, the mean concentration was 0.92 mg/liter and CL/metabolized fraction (f_m) 18.91 liters/h. Colistin concentrations were below the current MIC breakpoints, and the area under the concentration-time curve for the free, unbound fraction of the drug over 24 h in the steady state divided by the MIC (fAUC/MIC) was lower than recommended, suggesting that a dosage regimen of 160 mg CMS every 8 h (q8h) is inadequate.     

High prevalence of smoking in Northern Greece.

AIM: To investigate the prevalence of smoking in the general population and in specific population sub-groups in Northern Greece. METHODS: A cross-sectional study was conducted during the period 1999-2001 on a 5% sample (23,840) of those people aged between 21 to 80 out of a total general population of 653,249. 21,854/23,840 general population subjects were interviewed. In addition, we interviewed 9,276 high school students, 1,072 medical students, 597 medical doctors within the National Health System, 825 teachers, and 624 subjects who regularly exercised in a privately-owned gym. A specially modified ICRF study group questionnaire was used. RESULTS: 34.4% of the general population sample were current smokers (47.8% of males and 21.6% of females). Smoking prevalence rates in the population sub-groups were: 29.6% of high school students; 40.7% of medical students; 44.9% of medical doctors; 46.4% of teachers; and 36.9% of the gym group. CONCLUSION: The prevalence of smoking in Northern Greece is high. High school and medical students present with high smoking rates, and the same situation is observed in medical doctors and teachers. An intensification of preventive antismoking measures is required.     

Invasive cutaneous infection with Geotrichum candidum: sequential treatment with amphotericin B and voriconazole.

A rare case of an invasive cutaneous infection by Geotrichum candidum in an 80-year-old male patient with diabetes mellitus is reported. The primary site of infection manifested after trauma as an ulcerative lesion on the distal phalanx of the midfinger and extended throughout the right hand. Histological examination showed fungal invasion in the deep dermis without vascular involvement and G. candidum was grown in cultures from the biopsy material. Angiography revealed severe obstructive disease of the right brachial artery and its branches. Treatment, after susceptibility testing of the isolated strain, consisted of sequential administration of intravenous liposomal amphotericin B with oral voriconazole followed by liposomal amphotericin B, resulting in substantial improvement of the infection.     

Loss of ganglion cells in the retina secondary to vincristine therapy]

We report the case of a 25-year-old black female from Zaire with AIDS diagnosed 2 years earlier. Nine months before her death, she was treated for a disseminated Kaposi sarcoma with vincristin, adryamycin and bleomycin. At that time, visual acuity was normal and ophthalmologic examination was unremarkable except for the presence of bilateral Drusen and a cotton wool spot OS. Three months after the onset of chemotherapy, the patient complained of progressive visual field constriction, which progressed to blindness within a 4 month period. Five months after the onset of the tri-therapy a bilateral CMV retinitis developed, which was successfully treated by intravitreous injections of ganciclovir. This therapy was stopped as soon as blindness was established, with subsequent massive bilateral recurrence of the CMV retinitis. Histologic examination showed complete atrophy of the retinal ganglion cells and areas of CMV retinitis. The optic nerve was demyelinated and exhibited astrocytic gliosis. Immunohistochemistry confirmed the presence of CMV in infected retina and revealed the absence in the optic nerve of the class III beta-tubulin isotype and of the 200 kd neurofilament subunit. In contrast, oculomotor nerves appeared intact. The presence of HIV in the eye and in the optic nerve was excluded using PCR technique. The retinal ganglion cell loss and optic nerve atrophy appeared to be purely degenerative in nature, since there was no evidence of vascular occlusion, inflammation or retrobulbar compressive process. We therefore conclude that blindness was caused by vincristine therapy. The patient actually received 22 mg of vincristin intravenously in 11 courses over 7 months, although discontinuation was recommended by us after 5 months.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inhibition of allergic and non-allergic histamine secretion from rat peritoneal mast cells by calcium antagonists.

Bepridil, TMB-8 (8-(diethylamino)octyl 3,4,5-trimethoxybenzoate hydrochloride), diltiazem, verapamil and nifedipine exerted concentration-dependent inhibition of antigen and calcium ionophore A23187-induced histamine release from rat peritoneal mast cells. The inhibitory effects of verapamil and bepridil against calcium ionophore A23187-induced histamine secretion were antagonized by increased Ca2+ concentrations in the extracellular medium. These observations suggest that both agents act by interfering with the influx of Ca2+ into the mast cells. The inhibitory activities of five different Ca2+ channel blockers on allergic and non-allergic histamine secretion from rat peritoneal mast cells varied considerably depending upon the nature of the secretagogue as well as concentration and type of Ca2+-antagonist examined.     

Induction of non-specific airway hyperreactivity by potassium channel blockade in rat isolated trachea.

1. The exposure of rat isolated tracheal segment to the K(+)-channel inhibitor tetraethylammonium (TEA, 10 mM) for a period of 10-15 min generally produced little or no contractile response. 2. Cooling (10 C) provocation alone usually produced small (10 +/- 3% acetylcholine maximum) contractile responses. 3. In the presence of TEA (10 mM, 10-15 min exposure), rat trachea exhibited airway hyperreactivity to acetylcholine, 5-hydroxytryptamine (5-HT) and cooling. It also increased the peak tension induced by 5-HT. 4. TEA-induced airway hyperreactivity to cooling was significantly inhibited in Ca2(+)-free Krebs solution suggesting an important role for extracellular Ca2+ influx. 5. We conclude that the blockade of potassium channels with TEA induces non-specific airway hyperreactivity to cooling, 5-HT and acetylcholine in rat isolated tracheal segments.