Evidence that peroxynitrite affects human osteoblast proliferation and differentiation.

Peroxynitrite (PN), a nitric oxide (NO*)-derived anion, has been associated with NO* damage in various cell types. We examined the effects of adding PN to cultured human osteoblast-like (hOB) cells obtained after hip arthroplasty. Exposure to PN (0.1-0.4 mM) decreased both hOB proliferation and differentiation, measured by [3H]thymidine uptake and alkaline phosphatase production, respectively. Incubation with 3-morpholinosydnonimine (SIN-1; 0.25-1 mM), an NO* and O2- donor that leads to PN release, also reduced both hOB proliferation and differentiation. Coincubation with both superoxide dismutase (SOD; 100 U/ml) and catalase (CAT; 50 U/ml), rendering SIN-1 a pure NO* donor, reversed its effects on hOB proliferation and differentiation. However, SIN-1-induced NO* production, measured by nitrite release to the hOB medium, was not altered by cotreatment with SOD and CAT. Expression of nitrotyrosine by hOB, a marker of PN action, was significantly increased after SIN-1 addition, as compared with untreated cells, as revealed by Western blot analysis. Interleukin-1alpha (IL-1alpha) and interferon gamma (IFN-gamma) but not tumor necrosis factor alpha (TNF-alpha) also significantly increased nitrotyrosine expression in these cells. These data show that PN is at least partially responsible for osteoblast derangement by NO* and that cytokines released during inflammatory arthropathies can induce PN production in hOB cells.     

Serological diagnosis of infection by Shigella dysenteriae-1 in patients with bacillary dysentery

A total of 192 samples of serum from 113 Sri Lankan patients with clinical dysentery was examined for antibodies of the IgM class to the lipopolysaccharides (LPSs) of Shigella dysenteriae-1 and Escherichia coli O157:H7. By means of ELISA and immunoblotting, 59 patients were found to have serum antibodies to the LPS of S. dysenteriae-1 only. Four samples from one patient were found to contain serum antibodies to the LPSs of both S. dysenteriae-1 and E. coli 0157: H7. Antibodies to the LPS of S. dysenteriae-1 were also detected in 16 samples from 25 children, from Sri Lanka, with no previous history of dysentery; one of these children also had antibodies to the LPS of E. coli 0157: H7. Analysis of 16 samples from apparently healthy children in the U.K. showed that only one serum contained antibodies to the LPS of S. dysenteriae-1. This patient had a history of recent travel to Pakistan. The isolation of S. dysenteriae-1 remains the preferred test for the diagnosis of bacillary dysentery. The use of serology as a means of providing evidence of infection with S. dysenteriae-1, however may prove to be a useful adjunct to cultural techniques but needs to be validated in an area where this organism is endemic.     

Percutaneous selective thermorhizotomy in the treatment of "essential" trigeminal neuralgia. The importance of lesion selectivity

Results following percutaneous thermorhizotomy for trigeminal neuralgia are described in 111 patients. Recurrences and side effects are more frequent whenever selectivity of the surgical lesion has been imperfect (exceeding the original pain area and causing marked hypoesthesia), and less frequent in the cases with strictly selective lesion.     

Stimulation of enzyme secretion from isolated pancreatic acini by Clostridium difficile toxin B

Exposure of isolated rat pancreatic acini to increasing concentrations (10 ng - 800 ng/ml) of toxin B from Clostridium difficile produced a biphasic effect on the rate of secretion of amylase, trypsinogen, and chymotrypsinogen. Whereas doses of toxin B from 10-30 ng/ml increased enzyme secretion by 15-20%, doses between 30 ng and 60 ng/ml showed a regression of this effect, whereafter the rate of secretion of amylase, trypsinogen, and chymotrypsinogen increased with increasing concentrations of the toxin. Toxin B concentration of 800 ng/ml enhanced amylase, trypsinogen and chymotrypsinogen secretion by 119%, 185% and 195%, respectively, when compared with the basal level. Stimulation of enzyme secretion by toxin B was not affected by the presence of either actinomycin-D or cycloheximide, at a concentration which inhibited acinar RNA or protein synthesis by 80-90%. Although toxin B as well as CCK8, carbachol and secretin by themselves caused significant stimulation in amylase, trypsinogen and chymotrypsinogen secretion from isolated pancreatic acini, toxin B together with either CCK8, carbachol or secretin produced no further augmentation in enzyme secretion than what was observed with the secretagogues alone. It is concluded that toxin B of Cl. difficile exerts a direct effect on pancreatic acinar cells as evidenced by stimulation of enzyme secretion.     

Concentration and distribution pattern of selected micronutrients in preterm and term milk from urban Brazilian mothers during early lactation

The purpose of this study was to assess the concentration and binding pattern of zinc, iron, folate and vitamin B12 in milk of Brazilian women of low socioeconomic status giving birth at term or preterm, during early lactation. Protein, fat, total solids and ash concentrations were also determined. Protein and zinc concentrations decreased significantly as lactation proceeded whereas milk fat and folate increased with the lactation period. Total solids, ash, iron and vitamin B12 remained unchanged. Zinc was present mainly in the whey fraction while less than half of the total iron was present in this fraction, with no significant change due to stage of lactation. Unsaturated folate and vitamin B12 binding capacities and percentage of saturation of the folate binding protein increased with the stage of lactation. The vitamin B12 binding protein was highly unsaturated in all samples. There was no significant difference between term and preterm samples in the parameters investigated. Correlation analysis between milk components indicated significant relationships between total solids and fat, total zinc and whey zinc, folate and total folate binding capacity, unsaturated and total folate binding capacity, and unsaturated and total B12 binding capacity. In general, the nutrient concentrations found in this study are in good agreement with published data on milk composition of women from developed countries, with the possible exceptions of folate, which was lower, and iron, which was higher. The correlation of folate concentration with its binding protein found in this work supports the hypothesis of a regulatory role for folate levels in milk exerted by the folate binding protein in the mammary gland.