Homeostatic regulation of zinc transporters in the human small intestine by dietary zinc supplementation.

BACKGROUND: The role of intestinal transporter regulation in optimising nutrient absorption has been studied extensively in rodent and cell line models but not in human subjects. AIMS: The aim of the present study was to investigate the response in vivo of zinc transporters in the human enterocyte to dietary zinc supplementation. SUBJECTS: Eighteen patients who had previously undergone ileostomy, all free of any symptoms of inflammatory bowel disease. METHODS: Subjects took a daily zinc supplement of 25 mg for 14 days in a double blind, placebo controlled, crossover trial. The effect of the supplement on expression in ileal biopsies of the zinc transporters SLC30A1, SLC30A4, SLC30A5, SLC39A1, SLC39A4, and metallothionein was measured by reverse transcription-polymerase chain reaction RT-PCR. Expression of SLC30A1, SLC30A5, and SLC39A4 was also examined by immunoblotting. RESULTS: The zinc supplement reduced SLC30A1 mRNA (1.4-fold) together with SLC30A1, SLC30A5, and SLC39A4 protein (1.8-fold, 3.7-fold, and to undetectable levels, respectively) in ileal mucosa and increased metallothionein mRNA (1.7-fold). The supplement had no effect on expression of SLC30A4 or SLC39A1 mRNA. Localisation of SLC30A5 at the apical human enterocyte/colonocyte membrane and also at the apical membrane of Caco-2 cells was demonstrated by immunohistochemistry. Commensurate with these observations in zinc supplemented human subjects, SLC30A1, SLC30A5, and SLC39A4 mRNA and protein were reduced in Caco-2 cells cultured at 200 muM compared with 100 muM zinc. CONCLUSIONS: These observations indicate that, in response to variations in dietary zinc intakes, regulated expression of plasma membrane zinc transporters in the human intestine contributes to maintenance of zinc status.

Managing genital infection in community family planning clinics: an alternative approach to holistic sexual health service provision.

OBJECTIVE: To pilot and evaluate sexually transmitted infection (STI) management in community family planning clinics (FPCs). METHODS: Number of STI tests taken, positive results, infections treated, contacts traced/treated, referrals to specialist services and time from testing to treatment were documented as well as age and sex of the population tested. RESULTS: STI tests taken increased from 233 to 308/month and male clients seen increased from 114 to 147/month across all clinics. Chlamydia prevalence rates in one large clinic increased from 6.7% to 11.9%. 82% of those with STIs in this clinic were treated. Of 44 clients treated for chlamydia, 84% had partner notification performed, 0.43 contacts were treated for every client with chlamydia and referrals to specialist services decreased. 70% of STIs were detected in clinic users under the age of 25 and 45.5% of clients tested under the age of 16 had an STI. Before STI treatment was available at FP clinics 52% of clients with STIs attended specialist services after referral and time from testing to treatment was 19 days. Managing STIs in the community increased treatment rates to 82% with a testing to treatment time of 10 days. CONCLUSIONS: The management of uncomplicated genital infection in community FPCs working in partnership with specialist services is a feasible and effective approach to holistic sexual health service provision.

The effect of curcumin on glutathione-linked enzymes in K562 human leukemia cells.

Curcumin, an antioxidant present in the spice turmeric (Curcuma longa), has been shown to inhibit chemical carcinogenesis in animal models and has been shown to be an anti-inflammatory agent. While mechanisms of its biological activities are not understood, previous studies have shown that it modulates glutathione (GSH)-linked detoxification mechanisms in rats. In the present studies, we have examined the effects of curcumin on GSH-linked enzymes in K562 human leukemia cells. One micromolar curcumin in medium (16 h) did not cause any noticeable change in glutathione peroxidase (GPx), glutathione reductase, and glucose-6-phosphate dehydrogenase activities. Gamma-glutamyl-cysteinyl synthetase activity was induced 1.6-fold accompanied by a 1.2-fold increase in GSH levels. GSH S-transferase (GST) activities towards 1-chloro-2,4-dinitrobenzene, and 4-hydroxynonenal (4HNE) were increased in curcumin-treated cells 1.3- and 1.6-fold, respectively (P = 0.05). The GST isozyme composition of K562 cells was determined as follows: 66% of GST Pl-1, 31% of Mu class GST(s), and 3% of an anionic Alpha-class isozyme hGST 5.8, which was immunologically similar to mouse GSTA4-4 and displayed substrate preference for 4HNE. The isozyme hGST 5.8 appeared to be preferentially induced by curcumin, as indicated by a relatively greater increase in activity toward 4HNE. Immunoprecipitation showed that GPx activity expressed by GST 5.8 contributed significantly (approximately 50%) to the total cytosolic GPx activity of K562 cells to lipid hydroperoxides. Taken together, these results suggest that GSTs play a major role in detoxification of lipid peroxidation products in K562 cells, and that these enzymes are modulated by curcumin.

Persistent increases in basal cerebral metabolic activity induced by morphine sensitization.

To characterize the underlying neuroanatomic substrate of morphine (MS) sensitization, changes in the local cerebral metabolic rate for glucose (LCMRglu) were examined in 95 brain regions of male F-344 rats using the 2-deoxy-D-[1-14C]glucose method. The results of these experiments demonstrate that MS-induced sensitization is manifested by increases in basal metabolic activity that last for at least 6 days. Although changes in basal metabolic rate were found to be more extensive in the presence of conditioned cues, the increases in LCMRglu in nonconditioned sensitized rats indicate a basic underlying pharmacologic effect of MS sensitization on basal brain activity. Regions in which MS sensitization had a lasting pharmacologic effect include the shell of the nucleus accumbens, the prelimbic area of the prefrontal cortex, and the dorsolateral prefrontal cortex. Interestingly, the core of the nucleus accumbens and regions of the caudate were found to have an increased LCMRglu only in the presence of conditioned cues, indicating conditioned brain activity without observable changes in behavior. The previous administration of an MS-sensitizing treatment was also found to alter the cerebral metabolic response to a subsequent acute MS challenge (0.5 mg/kg, subcutaneously), most notably in forebrain systems. The more widespread activation of brain structures in the basal state in the presence of conditioned cues suggests that these MS-sensitized rats may model an altered brain state related to craving in the abstinent opiate addict.

Effect of dosing vehicle on the toxicity and metabolism of unsaturated aliphatic nitriles.

The effect of dosing vehicle on toxicity and metabolism of unsaturated aliphatic nitriles was investigated in male Sprague-Dawley rats. Five unsaturated aliphatic nitriles--acrylonitrile, methacrylonitrile, allylnitrile, crotononitrile and fumaronitrile--were prepared in five different dosing vehicles--saline, corn oil, safflower oil, mineral oil, olive oil and Tween-20. Groups of six male rats were given 0.5 LD50 doses of the nitriles by gavage and they were observed for 12 It for cholinomimetic and central nervous system effects. Cyanide and glutathione levels were determined in blood and various organs at 1, 3 and 6 h after nitrile administration and thiocyanate levels were determined at 6 h after nitrile administration. The results indicate that all the vehicles studied potentiated the toxicity of all the nitriles compared to nitriles administered in saline and significantly increased their metabolism to cyanide and thiocyanate and nitrile-induced depletion of glutathione in rats. This behavior of vehicles illustrates the difficulty of identifying suitable vehicles for administration of lipophilic compounds in toxicology studies.

Biofeedback: a new modality in the management of children with fecal soiling.

Fifty children and adolescents who had severe fecal incontinence associated with either imperforate anus surgery in infancy or longstanding functional constipation were given biofeedback training for the purpose of achieving anal sphincter control. Feedback was in the form of oscilloscope tracings which the children learned to produce by contracting small air-filled balloons positioned at the internal and external anal sphincters. Forty-seven of these patients learned to have voluntary bowel movements, and 30 eliminated soiling accidents completely during follow-up periods ranging from six months to three years.

Relationship between protein and ribonucleic acid synthesis during outgrowth of spores of Bacillus cereus.

The rate of protein and ribonucleic acid (RNA) synthesis was examined during the outgrowth of spores of Bacillus cereus T in a chemically defined medium. RNA synthesis started 2.5 min after the initiation of germination, and protein synthesis after 4 min. Addition of a complete amino acid supplement and uracil supported high rates of RNA and protein synthesis throughout outgrowth. To determine the relationship between the rate of protein (k) and RNA synthesis, the kinetics of formation of various classes of RNA were followed during outgrowth. Ribosomal RNA (rRNA) comprised a relatively constant fraction of the total RNA throughout outgrowth (71 to 78%). The classes of RNA synthesized during this period were determined by germinating spores in radioactive uracil and then at intervals following their stability to actinomycin D. Initially, labile RNA comprised the largest fraction of newly formed RNA (DeltaRNA), and this proportion decreased during outgrowth. The ratio of k/rRNA or k/Delta stable RNA varied considerably during outgrowth, whereas the ratio of k/labile RNA remained constant. The data suggest that the rate of protein synthesis is not rigidly coupled to either total or newly synthesized rRNA (ribosomes) during the early stages of outgrowth.