Rats' urinary metabolomes reveal the potential roles of functional foods and exercise in obesity management.

The complexity of the metabolic changes in obese individuals still presents a challenge for the understanding of obesity-related metabolic disruptions and for obesity management. In this study, a gas chromatography mass spectrometry (GC-MS) based metabolomics approach targeting urine metabolism has been applied to assess the potential roles of functional foods and exercise for obesity management in rats. Male albino rats diagnosed as obese via histopathology and biochemical assays were administered functional foods in common use for obesity management including pomegranate, grapefruit, and red cabbage juice extracts in parallel with swimming exercise. Urine samples were collected from these rats, and likewise from healthy control animals, for metabolite analysis using (GC-MS) coupled to multivariate data analysis. The results revealed a significant elevation in oxalate and phosphate levels in obese rat urine concurrent with lower lactate levels as compared to the control group. Furthermore, and to pinpoint the bioactive agents in the administered functional foods, ultra performance liquid chromatography (UPLC) coupled to high resolution time-of-flight mass spectrometry (TOF-MS) was employed for secondary metabolite profiling. The different phenolic classes found in the examined functional foods, viz. ellagitannins in pomegranate, flavanones in grapefruit and flavonols in red cabbage, are likely to mediate their anti-obesity effects. The results indicate that these functional foods and exercise were quite effective in reverting obesity-related metabolic disruptions back to normal status, as revealed by orthogonal partial least squares-discriminant analysis (OPLS-DA).

Evaluation of immunodiagnostic potential of ESAT-6 synthetic peptides mixture in Egyptian pulmonary tuberculosis patients.

Identification of M. tuberculosis ESAT-6 proteins has enhanced research for developing new specific and reliable in vitro diagnostic tests for pulmonary tuberculosis. The aim of this study was to assess the diagnostic potential of ESAT-6 synthetic overlapping peptides mixture (Pepmix) for diagnosis of pulmonary M. tuberculosis infection in Egypt. IFN-gammaand IL-17 production was assessed in whole-blood culture assay after overnight stimulation with M. tuberculosis crude culture filtrate (CF), purified 38 kDa antigen, and ESAT-6 Pepmix. Concentration of IFN-gamma and IL-17 was evaluated in the culture supernatant by ELISA. Patients with active pulmonary TB patients (n = 37), Tuberculin Skin Test (TST) positive healthy subjects (n = 16) and TST negative healthy subjects (n = 16) were included. Results indicated that IFN-gammaproduction following stimulation with crude CF, and 38 kDa purified antigen was not significantly different between patients and TST+ healthy controls. In contrast, Pepmix clearly distinguished between patients and TST+ healthy subjects with sensitivity of 70.3 % and specificity 96.9 %. The concentration of secreted IL-17 was significantly lower than IFN-gamma, and no statistically significant difference was found between groups. It is concluded that IFN-gamma assay using ESAT-6 Pepmix, as antigenic stimulus, has the potential to rapidly diagnose pulmonary TB infection.

Immunomodulatory effects of secondary hyperparathyroidism on circulating CD4+ and CD8+ T-lymphocytes in chronic renal failure patients.

The immunomodulatory effects of parathyroid hormone (PTH) in patients with end stage renal disease (ESRD) is controversial. This study was carried out to investigate the effect of PTH levels on the circulating CD4+, CD8+ T cell counts (%) in patients with chronic renal failure (CRF) on regular hemodialysis ((HD). The study included 22 patients with serum levels of PTH < 300 pg/ml (group 1), 18 patients with PTH > 300 pg/ml (group II) and 10 age and sex matched normal controls (group III). Chemiluminescence and flowcytometry assays were performed for determination of serum PTH levels and T cell subset counts respectively. The mean (%) of total lymphocyte, CD4+, CD8+ and CD4\CD8 ratio of group I were (81.68+/- 9.38), (52.00+/-6.24), (27.13+/- 6.31) and (1.99+/-0.42) respectively, as compared to (73.83+/-13.30), (46.05+/-8.59), (23.05+/-4.63) and (2.03+/-0.41) respectively in group II. Values of group I and II were significantly (P<0.001) lower than controls (88.50 +/- 6.02), (63.30 +/- 6.44), (36.80 +/- 6.44) and (1.76+/-0.36) respectively. In group II, the reduction was significantly (P<0.001) prominent in patients with high PTH levels, with significant inverse correlations (P<0.001) between PTH and % of total lymphocyte (r= -0.93), CD4+ (r= -0.74) and CD8+ % (r=-0.69). In conclusion, increased level of PTH in CRF patients on hemodialysis is associated with lymphopenia and reduction in CD4+ & CD8+ subsets of T cells. Monitoring circulating PTH levels in such patients can restore their immune competence.

Interferon gamma assay for diagnosis of childhood tuberculosis.

We measured interferon-gamma (IFN-gamma) in supernatant of peripheral blood mononuclear cell (PBMC) cultures stimulated with mycobacterial antigens. The study included ten children with newly diagnosed active pulmonary tuberculosis (TB), ten with inactive TB and 8 healthy controls. IFN-gamma was quantified by ELISA. Antigen stimulated PBMCs produced IFN-gamma at significantly higher levels than the background levels of nonstimulated cultures. Following specific antigen stimulation in vitro with TB bacilli, the PBMCs from patients with active and inactive TB produced significantly higher levels of IFN-gamma (433.15+/-119.80 and 420.64+/-131.52 respectively) than controls (241.2+/-119.74), (P< 0.001). However, no significant difference was found between active and inactive TB patients. In conclusion, estimation of specific IFN-gamma production by PBMC may be a useful immunological tool for diagnosis of childhood tuberculosis.

Serum antibodies to group A streptococcal extracellular and cell-associated antigens in Egyptians with post-streptococcal diseases.

We investigated serum antibodies to a comprehensive array of group A streptococcal antigens and superantigens in Egyptian subjects. Antibodies to Streptococcus pyogenes cell-associated proteins and to proteins released by rapidly dividing S. pyogenes were compared in four patient groups with different post-streptococcal diseases and in healthy controls. Enzyme-linked immunosorbent assays showed that total Ig and IgG to extracellular antigens were significantly higher in patients with acute rheumatic fever (ARF) compared to healthy controls, but no differences were found in either total Ig or IgG titres to cell-associated proteins between any of the groups. Western blotting showed that multiple extracellular and cell-associated antigens, covering a wide range of molecular masses, were recognised by all sera, including healthy controls. No evidence was obtained for putative dominant antigens associated with any disease group, although a low molecular mass cell-associated protein (approximately 4 kDa) was clearly recognised by two-thirds of subjects irrespective of disease status. These findings demonstrate that raised serum Ig and IgG titres to extracellular, but not cell-associated, S. pyogenes antigens are a feature of ARF in this population, and suggest that multiple S. pyogenes antigens contribute to this response.

HBeAg in viral hepatitis type B.

58 patients with acute hepatitis type B, including 13 with fulminant hepatitis, were tested on presentation for HBeAg. Positive results were obtained in 24%. The frequency was highest in those with fulminant hepatitis (46% positive), and this was probably related to the earlier presentation of patients with this condition. Patients with acute hepatitis who were HBeAg-positive had had a significantly shorter duration of symptoms than those who were HBeAg-negative at presentation. HBeAg was still detectable more than 3 weeks after the onset of symptoms in only 2 patients, both of whom progressed to chronic active hepatitis. The early detection of HBeAg in patients with acute hepatitis is of no prognostic significance, but its persistence may provide the earliest evidence of potential chronicity.

Enhanced HBsAb production in pathogenesis of fulminant viral hepatitis type B.

The possible importance of humoral immunity in the pathogenesis of fulminant hepatitis was investigated by comparing 17 patients with fulminant hepatitis type B with 20 patients with severe but non-fulminant disease. Hepatitis B surface antigen (HBsAg) was cleared from the serum significantly faster (P less than 0-001) in those with fulminant hepatitis, and in 41% anti-HBsAg (HBsAb) was detectable by radioimmunoassay (RIA) at presentation. In all 11 sera from patients with fulminant hepatitis that were examined by electron microscopy aggregates of HBsAg and HBsAb were seen. In contrast, HBsAb was never detected by RIA in those with non-fulminant hepatitis, and in only one serum specimen (5%) were aggregates seen on electron microscopy. A significant sex difference between fulminant and non-fulminant hepatitis was observed, 65% of patients with fulminant hepatitis but only 15% of patients with non-fulminant hepatitis being women (P less than 0-01). An enhanced production of HBsAb in fulminant hepatitis, by leading to free HBsAb in portal blood, may cause an Arthus reaction in the sinusoids of the liver with ensuing ischaemic necrosis of hepatocytes.

Immune response to HBsAg and the spectrum of liver lesions in HBsAg-positive patients with chronic renal disease.

Evidence of chronic hepatitis was found on histological examination in nine out of 15 patients positive for hepatitis-B surface antigen (HBsAg) who had either chronic renal failure or a functioning renal transplant. Cirrhosis had already developed in three of the patients, who deteriorated rapidly and died. Liver biopsies from the remaining 12 patients showed the features of chronic aggressive hepatitis in two, chronic persistent hepatitis in four, and minor histological lesions in six. The persistence of HBsAg in patients with renal failure or in those receiving immunosuppressive drugs after a transplant must indicate some impairment of the normal immune response to hepatitis-B viral antigens. Nevertheless, cellular or humoral immunity to HBsAg was detected in all eight patients with chronic hepatitis tested compared with only one out of five with minimal liver lesions, which suggests that the severity of the liver damage may be directly related to the degree of immunocompetence.

e Antigen-antibody system as indicator of liver damage in patients with hepatitis-B antigen.

The clinical relevance of the e antigen-antibody system was investigated in 61 people persistently positive for hepatitis-B surface antigen, including 22 healthy carriers. The e antigen was not detectable in any of the healthy carriers, whereas it was found in 15 out of 28 patients with chronic aggressive hepatitis and two out of 11 with chronic persistent hepatitis. Its presence therefore indicates chronic liver disease but its absence does not exclude it. It may prove to be a particularly useful prognostic aid in chronic persistent hepatitis, since one of the two patients in whom it was found later developed aggressive hepatitis. In contrast, e antibody is of little diagnostic help, for, though it was found mostly in healthy carriers (18;82%), it was also detectable in 9 (23%) of the patients with chronic hepatitis. In 13 (76%) of the patients positive for e antigen Dane particles were seen on electron microscopy, but these were also present in 5 (19%) of the patients positive for e antibody. These findings are consistent with other evidence suggesting that e antigen is not a surface component of the Dane particle, but rather an independent soluble protein manufactured by the host in response to infection with the hepatitis-B virus.