Urine proteome analysis to evaluate protein biomarkers in children with autism.

BACKGROUND: Autism is a complex developmental disability for which no specific diagnostic markers have been identified so far. The present study aimed to evaluate whether there is any abnormal protein(s) excreted in the urine of autistic children by proteome analysis which may act as diagnostic marker. METHODS: Urine proteome analysis was carried out in first void urine samples of autistic and normal children (n=30) in the age group of 4-12 years by 2D-PAGE followed by MALDI-TOF-MS analysis. RESULTS: Comparison of 2D-PAGE gels revealed that many urinary proteins are expressed differentially in autistic children. Total numbers of spots observed were 250 and 159 in autism and normal samples respectively, out of which 95 matches were observed. In addition, 3 spots of abnormally expressed peptides were selected, excised and analyzed. Peptide sequence with significant match score was for kininogen-1 (KNG-1)-50 (spot-1), IgG1 heavy chain variable region-35(spot-2) and mannan-binding lectin serine protease-2 isoform-2 precursor-45(spot-3). All the autistic children showed significant increase (p<0.001) in urinary kininogen level measured quantitatively by ELISA, when compared to normal children. CONCLUSION: Increased urinary kininogen-1 level in all the autistic children and the possibility of this protein as a diagnostic marker need further investigation.

Rutin rich Emblica officinalis Geart. fruit extract ameliorates inflammation in the pancreas of rats subjected to alcohol and cerulein administration.

BACKGROUND: The modulating effect of methanolic extract of Emblica officinalis (MEEO) on ethanol (EtOH)- and cerulein (Cer)-induced pancreatitis in rats was investigated in this study. METHODS: Male albino Wistar rats were divided into four groups. Group 1 and 2 rats served as control and fed normal diet. Group 3 and 4 rats were fed isocalorically adjusted diet containing EtOH (36% of total calories) for 5 weeks and also subjected to intraperitoneal injection of Cer 20 µg/kg b.wt. thrice weekly for the last 3 weeks of the experimental period. In addition, group 2 and 4 rats received 200 mg/kg b.wt. of MEEO from 15th day till the experimental period. Serum levels of lipase (L), amylase (A), cytokines IL-1ß, IL-18, caspase-1 and oxidative stress index (OSI) were determined. Levels of fecal trypsin, total collagen, caspase-1, myeloperoxidase (MPO), antioxidants and mRNA expression of caspase-1, IL-1ß and IL-18 were determined in the pancreas. RESULTS: HPLC analysis showed the presence of rutin in MEEO. We observed a significant elevation in serum L/A ratio, IL-1ß, IL-18, caspase-1, OSI, collagen, MPO activity and the mRNA expression of IL-1ß, IL-18 and caspase-1 and significant reduction in fecal trypsin and antioxidant status in EtOH- and Cer-administered rats. The inflammatory markers were found to be reduced and the antioxidant status of pancreas was maintained in MEEO-coadministered rats. CONCLUSIONS: The rutin rich nature of E. officinalis can be claimed for its anti-inflammatory and pancreato protective effects.

Abnormal circadian rhythm and cortisol excretion in autistic children: a clinical study.

AIM: To determine the circadian rhythm alteration of cortisol excretion and the level of corticosteroids in children with different grades of autism severity. METHODS: The study included 45 children with different grades of autism severity (low [LFA], medium [MFA], and high functioning autism [HFA]), 15 in each group, and 45 age/sex-matched children with typical development. The urinary levels of free cortisol (at three phases of 24-hour cycle), corticosteroids, vanilylmandelic acid, and 5-hydroxyindole acetic acid were determined. RESULTS: Alteration in the pattern of cortisol excretion (Phases I, II, and III) was observed in children with LFA (Phase I: 43.8 ± 4.43 vs 74.30 ± 8.62, P=0.000; Phase II: 21.1 ± 2.87 vs 62 ± 7.68, P<0.001; Phase III: 9.9 ± 1.20 vs 40 ± 5.73, P<0.001) and MFA (Phase I: 43.8 ± 4.43 vs 52.6 ± 7.90, P<0.001; Phase II: 21.1 ± 2.87 vs 27.4 ± 4.05, P<0.001; Phase III: 9.9 ± 1.20 vs 19 ± 2.50, P<0.001) compared to the control group. The corticosteroids excretion levels were higher in all the groups of children with autism than in the control group. The level of 5-hydroxyindole acetic acid was significantly higher in children with LFA (8.2 ± 1.48 vs 6.8 ± 0.85, P<0.001) and MFA (8.2 ± 1.48 vs 7.4 ± 0.89, P=0.001) and not significantly higher in children with HFA than in the control group. The changes were correlated with degrees of severity of the disorder. CONCLUSION: These data suggest that altered cortisol excretion pattern and high level of corticosteroids in urine may probably be a consequence of altered hypothalamic-pituitary-adrenal axis function, which may contribute to the pathogenesis and affect the severity of autism.