Depressed Immune Responses and Accelerated Splenic Apoptosis due to Experience of Food Deprivation and Inequality but not Unstable Social Status in Balb/c Mice.

OBJECTIVE(S): We aimed to show that the immune system is sensitive to the detrimental effects of inequality and social injustice, and splenic vulnerability to apoptosis may also increase. METHODS: In order of better determination of immune responses to chronic social stress, we implemented food deprivation, food intake inequality, and unstable social status (a change of cage-mate every 3 days) for a period of 14 days in 60 male Balb/c mice. At the end of this stress period, nitric oxide (NO) production by peritoneal adherent cells and the serum concentration of corticosterone were measured. Moreover, the viability of peritoneal adherent cells and spleen lymphocytes was evaluated by MTT assay. The terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay was done to reveal the TUNEL-reactive apoptotic bodies in the spleen. RESULTS: Our results showed that food deprivation and inequality caused significant changes in the apoptosis of splenic cells in comparison with the control group (p < 0.05). Moreover, the vital activities of lymphocytes and peritoneal adherent cells, as well as NO production by the latter, increased significantly (p < 0.05). However, the experience of unstable social status did not cause a further increase in the viability of lymphocytes and peritoneal adherent cells, or NO production in animals that were food-deprived or experienced inequality. Serum concentration of corticosterone in all experimental groups, except for animals that experienced unstable social status only, significantly decreased versus the control group (p < 0.05). CONCLUSIONS: The results suggest that poverty and social inequality, but not unstable social status, affect immune responses and are likely involved in the induction of splenic apoptosis in mice.

Sewage as a rich source of phage study against Pseudomonas aeruginosa PAO.

Pseudomonas aeruginosa is a ubiquitous organism which has emerged as a major public health threat in hospital environments. Overuse of antibiotics has significantly exacerbated the emergence of multi-drug resistant bacteria such as P. aeruginosa. Phages are currently being utilized successfully for aquaculture, agriculture and veterinary applications. The aim of this study was to isolate and characterize of lytic P. aeruginosa phage from sewage of Ilam, Iran. Phage was isolated from sewage that was added to the enrichment along with the host and subsequently filtered. Plaque assay was done by using an overlay method (also called the double agar layer method). Purified plaques were then amplified for characterization. Finally, RAPD-PCR method was conducted for genotyping and Transition electron micrograph (TEM) recruited to determine the morphology and phage family. The phage had high concentration and tremendous effects against a variety of clinical and general laboratory strains (ATCC15693) of P. aeruginosa. Among a set of primers in RAPD panel, only P2 and RAPD5 primers, were useful in differentiating the phages. TEM images revealed that the isolated phages were members of the Siphoviridae family. The phage effectiveness and specificity towards target bacteria and potential to control biofilm formations will be investigate in our further studies.

Experience at a stricture clinic in a developing country.

OBJECTIVE: To evaluate the experience of 37 years regarding etiology, complications and evolving practice of the management of stricture urethra patients at the Sindh Institute of Urology and Transplantation, Karachi. METHODS: The retrospective descriptive study included 1600 cases of stricture urethra admitted to the Urology Section of the institute from 1972 to 2009. Files of all patients were reviewed; age, gender, site of stricture, etiology, diagnostic methods, clinical symptoms and management, as well trends and patterns were noted. RESULTS: There were 1600 patients of whom 1595 (99.4%) were males and 5 (0.4%) were females with age ranging from 14-80 years. Pelvic fracture urethral disfraction defects (n = 655; 49.5%) and fall astride (n = 123; 9.2%) were the commonest causes. Of the total, 92 percent presented with retention of urine. In the first decade, rail-road and dilatation was the mainstay of treatment. In the second decade, rail-road, dilatation and direct visual internal urethrectomy were the mainstay, but in the last 5 years, urethroplasty replaced the old methods. CONCLUSION: Trauma is the main cause of stricture. Over the years, urethroplasty and direct visual internal urethrotomy are the mainstay of management. Stricture clinic plays an important role in the early diagnosis of complications of stricture urethra and in the rehabilitation of these patients.

Management of stone disease: 17 years experience of a stone clinic in a developing country.

OBJECTIVE: To review 17 years experience of the stone clinic with incorporating the changes in practice over the years and to report the benefits of stone clinic in a developing country. METHODS: The SIUT Stone clinic was established in 1990 with installation of HM4 Lithotriptor. This clinic is run jointly by a Urologist, Dietitian, Nephrologist, Biochemist and Radiologist. From 1990 - 2007, about 38,749 stone patients received treatment with ESWL (55%), PCNL (6.0%), URS (15.5%), litholopaxy 4.0% and open surgery 19.7%. These patients after treatment were followed in the stone clinic with stone analysis and 24 hours urine metabolic studies where indicated. Dietary and oral hydration programme combined with medical therapy was also instituted. Recurrence rate was noted in those patients who were advised diet modification, oral hydration and medical treatment. Complications of stone disease were documented during the follow-up period. RESULTS: In ESWL group 8226 patients were followed in the stone clinic for 5 years. In this group, 185 (2.2%) had recurrence of renal calculi. In PCNL group 1306 patients were followed, and 16 (1.1%) had recurrence. In the open surgery group of 1294 patients, 17 (1.3%) had recurrence of renal calculi. The complications of stone disease noted were renal failure and infections in 162 (1.5%) patients, during the follow-up period. CONCLUSION: This study shows the beneficial effect of a stone clinic in a developing country and positive effect on the reduction of recurrence and complications of stone disease. Our experience promotes the need to open more specialized stone clinics in areas where stone disease is highly prevalent.

Dynamic responses to leptin secretagogues in lean, obese, and massively obese men and women.

BACKGROUND/AIM: Basal plasma leptin levels are higher in women than in men and also higher in obese than in lean subjects, but the dynamic leptin secretion has not been well studied. We tested whether the leptin secretory response to glucocorticoid or insulin differs by gender and adiposity status. METHODS: Seventy-nine nondiabetic adults, comprising lean [body mass index (BMI; kg/m(2)) < or =25; n = 27], obese (BMI 30-40; n = 28), and massively obese (BMI >40; n = 24) subjects, participated in two separate studies. In study 1, the subjects received oral dexamethasone (4 mg), with blood sampling before and 8 and 16 h after ingestion. In study 2, the subjects underwent a two-step hyperinsulinemic (1.0 mU.kg(-1)/min for 3 h, then 2.0 mU.kg(-1)/min for 3 h), euglycemic (approximately 100 mg/dl) clamp. Blood samples were obtained at baseline and every 20 min during the clamp. RESULTS: Basal and stimulated leptin levels were higher in women than in men, and higher in the obese groups than in lean subjects. The percentage increase above baseline leptin was similar among men and women within each group, but was approximately 30% lower in massively obese compared to lean subjects. CONCLUSION: Leptin secretory responses to glucocorticoid or insulin stimulation are preserved across a broad adiposity range, with higher absolute responses in women than in men.

Cellular and tissue localization of globotriaosylceramide in Fabry disease.

The pathogenesis of Fabry disease is poorly understood. We used a variety of immunohistological techniques to localize globotriaosylceramide, the main glycolipid that accumulates in Fabry disease. Globotriaosylceramide immunoreactivity in a heterogenous pattern was present in all organs examined of a patient on long-term enzyme replacement therapy. In the brain, immmunopositivity was found only in the parahippocampal region. Globotriaosylceramide immunostaining was present in the cell membrane and cytoplasm of endothelial cells, even in the absence of lysosomal inclusions. In kidney tissue, globotriaosylceramide colocalized with lysosomal, endoplasmic reticulum, and nuclear markers. Pre- and postembedding immunogold electron microscopy of skin biopsies and untreated patient cultured skin fibroblasts confirmed the presence of globotriaosylceramide in the cell membrane, in various cytoplasmic structures, and in the nucleus. Control organ tissues and cultured fibroblasts from five unaffected subjects were uniformly negative for globotriaosylceramide by immunohistochemistry and immunogold electron microscopy. We conclude that a substantial amount of lysosomal and extralysosomal globotriaosylceramide immunoreactivity remains in cells and tissues even after years of enzyme replacement therapy in Fabry disease. These findings are crucial for the understanding of the disease mechanism and suggest the usefulness of immunostaining for globotriaosylceramide as a means to assess response to novel, specific therapies.

The cerebral vasculopathy of Fabry disease.

Fabry disease is an X-linked disorder affecting both males and females. It is associated with an increased risk of stroke in up to 4% of patients below 55 years of age in the general population. The cerebral vasculopathy consists of ischemic strokes involving large and small vessels. The neuronal accumulation of glycosphingolipids appears to have no clinical effect on the natural history of Fabry disease with the possible exception of some reported mild cognitive abnormalities. The pathogenesis of Fabry vasculopathy remains poorly understood but is associated with abnormal functional control of the vessel secondary to endothelial dysfunction, cerebral hyper-perfusion and a prothrombotic state with likely increased production of reactive oxygen species. These abnormalities are further modified by genetic and possibly other vascular risk factors. This vasculopathy illustrates the role of glycolipids in this and possibly other types of cerebral vasculopathies. Therapy is preventive relying on standard medical care and in particular on anti-platelet agents such as clopidogrel.

Pathological findings in a patient with Fabry disease who died after 2.5 years of enzyme replacement.

We describe the postmortem findings of a 47-year-old man with Fabry disease, an X-linked glycolipid storage disorder, who was on enzyme replacement therapy with recombinant alpha-galactosidase A for more than 2 years. The patient had widespread atherosclerotic coronary artery disease that culminated in a massive acute myocardial infarction. Atherosclerotic lesions were seen in the right and left coronary systems, aorta, and the basilar artery. Typical Fabry cardiomyopathy and glomerular nephropathy were found. With the exception of vascular endothelial cells, extensive glycolipid storage deposits were seen in all vascular and nonvascular cells and organ systems. We conclude that, at least in this patient, repeated infusions with alpha-galactosidase A over a prolonged period did not appreciably clear storage material in cells other than vascular endothelial cells. These findings also illustrate accelerated atherosclerosis in susceptible patients with Fabry disease.

Energy adaptation to glucocorticoid-induced hyperleptinemia in human beings.

Recombinant leptin therapy potently decreases food intake and body weight in aleptinemic individuals with leptin gene mutations. However, it is unknown whether manipulation of endogenous leptin secretion alters ingestive behavior in otherwise healthy subjects. We therefore assessed energy consumption during administration of hydrocortisone (HC), a known leptin secretagogue. Six healthy adults were admitted overnight on 2 occasions and given HC (12.5 mg/h IV) or saline infusion for 24 hours (8:00 am -8:00 am ) in a randomized crossover design. Total energy and macronutrient intake was calculated using a computerized nutrient analysis program. Blood sampling for measurement of leptin, cortisol, glucose, and insulin was performed at baseline and every 1 to 2 hours. A rise in plasma leptin level was noted after approximately 5 hours of HC infusion and was sustained throughout the study period. The total energy consumed was 3004 +/- 231 kcal for saline and 2486 +/- 214 kcal for HC ( P = .005); breakfast energy values on day 1 were similar but energy values consumed at lunch, dinner, and day 2 breakfast were all significantly lower during induced hyperleptinemia. Analysis of macronutrients indicated significant decreases in carbohydrate and fat intake during glucocorticoid-induced hyperleptinemia as compared with placebo. These results indicate that stimulation of native leptin secretion decreases energy consumption, similar to the effect observed with recombinant leptin therapy. To our knowledge, this is the first report documenting anti-ingestive responses to a leptin secretagogue. Because chronic glucocorticoid therapy is fraught with adverse effects, we suggest that nonsteroidal or steroidomimetic leptin secretagogues may be good candidates for anorectic and antiobesity drug development.

Antisecretory effect of hydrogen sulfide on gastric acid secretion and the involvement of nitric oxide.

The present study was designed to investigate the effect of H2S on distention-induced gastric acid secretion. Fifty-two rats were randomly assigned to seven experimental groups. The gastric acid secretion was stimulated by gastric distention. Two groups of rats received L-cysteine or saline for 5 days before stimulation of the gastric acid secretion. Two groups of animals also received NaHS or saline just prior to stimulation of the gastric acid secretion. The effect of L-NAME and propargylglycine was also investigated. The mucosal levels of the gene expression of cyclooxygenase-2 (COX-2), endothelial nitric oxide synthase (eNOS), and H(+)/K(+)-ATPase α-subunit were quantified by qPCR and luminal concentrations of NO were determined. NaHS and L-cysteine decreased the gastric acid output in response to distention. The mRNA expression of H(+)/K(+)-ATPase α-subunit decreased by NaHS and L-cysteine as compared with the control group while gene expression of eNOS and COX-2 was upregulated. The inhibitory effect of NaHS on distention-induced gastric acid secretion was mitigated by pretreatment of L-NAME. These findings suggest the involvement of NO in mediating the antisecretory effect of H2S.

Comparison between the Ability of Glasgow Coma Scale and Full Outline of Unresponsiveness Score to Predict the Mortality and Discharge Rate of Pediatric Intensive Care Unit Patients.

OBJECTIVE: Prediction of survival and mortality rates in costly environments such as the intensive care unit (ICU) is of great importance for the assessment of new treatments, resource consumption control, and improvement of quality control. This study aimed to determine the ability to predict mortality and discharge rate of patients using the FOUR score in the pediatric ICU (PICU) of Ali Ibn Abitalib Hospital, Zahedan and compare the results with those of Glasgow Coma Scale (GCS). METHODS: This prospective study was conducted on 200 patients admitted to the PICU. Convenience purposive sampling was used. Research data was collected using the Full Outline of Unresponsiveness (FOUR) score and GCS using questionnaires. Obtained data was analyzed with SPSS 16 using descriptive statistics and correlation analyses. FINDINGS: Of the 200 children admitted to the PICU, 71.5% and 28.5% were discharged and died, respectively. The inter-rater reliability for the FOUR score was good to excellent (weighted κ: eye, 0.72; respiration, 0.82; brainstem, 0.74; motor, 0.78), In terms of mortality and discharge prediction, logistic regression analyses (FOUR score = OR: 0.13; 95% CI: 0.06-0.29; P<0.001; GCS=OR: 2.49; 95% CI: 1.44-4.32; P<0.001) showed that the FOUR score is a good predictor for in-hospital mortality. CONCLUSION: Results indicated that the FOUR score is more capable than GCS in predicting the mortality and discharge of patients admitted to the PICU.

Fasting plasma leptin level is a surrogate measure of insulin sensitivity.

CONTEXT: Published studies indicate marked variability in plasma leptin levels among persons with similar body mass index (BMI). We tested the hypothesis that such variations in leptin levels reflect differences in insulin sensitivity. SUBJECTS AND METHODS: Using euglycemic clamp, we assessed insulin sensitivity (ISI) in 57 nondiabetic adults (36 women, 21 men), whose BMI ranged from 20 to 78 kg/m2. We identified 38 age-matched subjects, stratified by fasting leptin (normal, <15 ng/ml vs. high, >or=15 ng/ml) and BMI (nonobese, <27 kg/m2 vs. overweight/obese, BMI>or=27 kg/m2) and compared ISI across the four strata. RESULTS: Fasting leptin levels correlated with ISI (r=-0.66 in men and -0.60 in women). In a multivariate regression model, leptin emerged as a strong predictor of ISI (r=-0.41, P=0.0002) after controlling for adiposity, whereas insulin weakened as a predictor (r=-0.32, P=0.0116). From regression plots of ISI vs. BMI and leptin, a BMI greater than 27 kg/m2 and a leptin level greater than 15 ng/dl strongly predicted decreased ISI. A fasting leptin cutoff of 15 ng/ml for detection of insulin sensitivity has a sensitivity of 72.7%, specificity of 56.3%, and positive predictive value of 69.6%. Overweight/obese subjects with fasting leptin less than 15 ng/ml were 100% more insulin sensitive than control subjects with leptin greater than 15 ng/ml. CONCLUSIONS: Insulin sensitivity explains about 40% of the variance in fasting leptin levels. Thus, fasting plasma leptin levels probably serve as an endogenous response to ambient insulin resistance and may provide a surrogate measure of insulin action.

Glucoregulatory physiology in subjects with low-normal, high-normal, or impaired fasting glucose.

OBJECTIVE: We determined whether variations in fasting plasma glucose (FPG) within the nondiabetic range represent differences in insulin action or secretion. SUBJECTS AND METHODS: Using results of the 75-g oral glucose tolerance test, we classified 39 adults (body mass index range 20-56 kg/m2) as having normal fasting glucose (NFG, <100 mg/dl, n = 24) or impaired fasting glucose (IFG, 100-125 mg/dl, n = 15). The NFG group was subdivided into low-NFG (<90 mg/dl, n = 11) and high-NFG (90-99 mg/dl, n = 13). The 2-h oral glucose tolerance test plasma glucose value was used to assign normal or impaired glucose tolerance (IGT) status. Insulin sensitivity was assessed by hyperinsulinemic euglycemic clamp, quantitative insulin sensitivity check index, and homeostasis model assessment of insulin resistance; beta-cell function was assessed by calculating the insulinogenic index, homeostasis model assessment, and the disposition index. RESULTS: Compared with low-NFG subjects, insulin sensitivity and glucose tolerance were lower among persons with isolated IFG and combined IFG-IGT. The hyperinsulinemic euglycemic clamp (micromol/kg x min(-1)/pmol/liter) was 0.109 +/- 0.011 in low-NFG subjects, 0.088 +/- 0.009 in IFG subjects (P = 0.04), and 0.022 +/- 0.003 in IFG-IGT subjects (P = 0.0014). The spread in FPG from 70-125 mg/dl was associated with a greater than 3-fold difference in insulin sensitivity. Compared with low-NFG subjects, the disposition index decreased by 32.8% in high-NFG subjects (P = 0.08), by 45.6% in subjects with isolated IFG (P < 0.02), and by 49.8% (P < 0.02) in those with combined IFG-IGT. CONCLUSION: Compared with persons with low-NFG, those with IFG or combined IFG-IGT have significant alteration of glucoregulatory physiology, whereas high-NFG (pre-prediabetes) status might portend nascent glucoregulatory perturbations.

Weekly enzyme replacement therapy may slow decline of renal function in patients with Fabry disease who are on long-term biweekly dosing.

This study was performed to determine whether adult male patients with Fabry disease who demonstrate a continuing decline in renal function despite 2 to 4 yr of conventionally dosed agalsidase alfa therapy (0.2 mg/kg every other week [EOW]) show an improved slope of decline with weekly administration using the same dosage. Eleven (27%) of 41 adult male patients with Fabry disease who participated in long-term agalsidase alfa clinical trials and who had demonstrated a slope of decline in estimated GFR (eGFR) of > or =5 ml/min per 1.73 m(2)/yr while receiving long-term treatment with agalsidase alfa at the currently recommended dosage of 0.2 mg/kg, infused EOW, were enrolled in this open-label, prospective study. Patients were switched from EOW to weekly infusions and followed for an additional 24 mo. Before switching to weekly dosing, eGFR was 53.7 +/- 6.3 ml/min per 1.73 m(2) (mean +/- SEM), and mean rate of change in eGFR was -8.0 +/- 0.8 ml/min per 1.73 m(2)/yr. During the 24-mo follow-up period after switching to weekly dosing, the mean rate of change in eGFR was observed to slow to -3.3 +/- 1.4 ml/min/1.73 m(2)/yr (P = 0.01 versus EOW). After switching to weekly dosing, three patients demonstrated an improvement in eGFR and six patients demonstrated a slowing in the rate of eGFR decline; only two patients failed to improve their eGFR slope. A multiple regression model confirmed that the weekly infusion regimen was the strongest explanatory variable for the change in eGFR (P = 0.0008), with a weaker contribution from the concomitant use of angiotensin converting enzyme inhibitors/angiotensin receptor blockers (P = 0.02). These results suggest that weekly infusions of agalsidase alfa at a dosage of 0.2 mg/kg may be beneficial in the subgroup of patients who have Fabry disease and whose kidney function continues to decline after 2 to 4 yr or more of standard EOW dosing.

Leptin response to glucocorticoid occurs at physiological doses and is abolished by fasting.

OBJECTIVE: To examine the effects of graded doses of hydrocortisone (HC) on leptin secretion, and determine the effect of fasting. RESEARCH METHODS AND PROCEDURES: This was a randomized, placebo-controlled, crossover study, with a 1-week "washout" period between interventions. Eight healthy subjects [age = 36 +/- 2.3 years (+/-SE), body mass index = 31.5 +/- 1.6 kg/m(2)] completed the dose-response study in which an intravenous infusion of saline (placebo) or HC (30 or 100 mg) was administered for 24 hours. Four healthy subjects (age = 35.2 +/- 3.0 years, body mass index = 27.1 +/- 2.1 kg/m(2)) completed the fasting study, which entailed continuous infusion of saline, HC (300 mg/24 hours) in the fed state, or HC (300 mg/24 hours) with total caloric deprivation for 24 hours. Blood sampling was performed every 1 to 2 hours for measurement of leptin, cortisol, insulin, and glucose levels. RESULTS: Peak hyperleptinemia occurred after 16 hours of HC infusion; peak/baseline leptin levels were 129% (placebo), 140% (30 mg of HC for 24 hours, p = 0.05), and 185% (100 mg of HC for 24 hours, p < 0.01). During infusion of HC (300 mg/24 hours or placebo), the peak/baseline plasma leptin levels were 16.1 +/- 5.8/12.8 +/- 5.9 ng/mL (placebo with food, 126%), 14.6 +/- 6.0/12.5 +/- 6.5 ng/mL (HC fasting, 117%), and 32.5 +/- 12.5/12.0 +/- 8.4 ng/mL (HC with food, 271%, p < 0.001). DISCUSSION: Leptin secretory responses occur at physiological doses of HC, are obliterated by fasting, and thus may be of metabolic significance.