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

An automated algorithm for the quantification of hCG level in novel fabric-based home pregnancy test kits

We report a new image processing algorithm that extracts quantitative information about the concentration of human chorionic gonadotropin (hCG), an important early pregnancy marker, from commercially available qualitative home pregnancy kits. The algorithm could potentially be ported onto a simple camera based cell phone making it a low-cost, portable point-of-care device as opposed to costly and time consuming clinical labs for accurate quantitative determination of hCG. The algorithm takes the image of the test result as input, classifies and determines the hCG concentration based on the RGB intensities of the test line. The efficacy of the algorithm is demonstrated using control samples on commercially available strips as well as novel fabric based strips designed for this application

Advanced Development of Primary Pancreatic Organoid Tumor Models for High-Throughput Phenotypic Drug Screening.

Traditional high-throughput drug screening in oncology routinely relies on two-dimensional (2D) cell models, which inadequately recapitulate the physiologic context of cancer. Three-dimensional (3D) cell models are thought to better mimic the complexity of in vivo tumors. Numerous methods to culture 3D organoids have been described, but most are nonhomogeneous and expensive, and hence impractical for high-throughput screening (HTS) purposes. Here we describe an HTS-compatible method that enables the consistent production of organoids in standard flat-bottom 384- and 1536-well plates by combining the use of a cell-repellent surface with a bioprinting technology incorporating magnetic force. We validated this homogeneous process by evaluating the effects of well-characterized anticancer agents against four patient-derived pancreatic cancer KRAS mutant-associated primary cells, including cancer-associated fibroblasts. This technology was tested for its compatibility with HTS automation by completing a cytotoxicity pilot screen of ~3300 approved drugs. To highlight the benefits of the 3D format, we performed this pilot screen in parallel in both the 2D and 3D assays. These data indicate that this technique can be readily applied to support large-scale drug screening relying on clinically relevant, ex vivo 3D tumor models directly harvested from patients, an important milestone toward personalized medicine

Characterization of soluble FAS, FAS ligand and tumour necrosis factor-alpha in patients with chronic HCV infection

Background: There are limited reports on the role of the cell surface receptor Fas and its ligand molecule in mediating apoptosis during infection with the hepatitis C virus (HCV). Objectives: The aims of this study were (1) to assess the susceptibility of the Fas antigen expressed on peripheral blood mononuclear cells to Fas ligand-induced-death in patients with chronic HCV infection and (2) to investigate the correlation between the plasma levels of soluble Fas (sFas), soluble Fas ligand (sFasL), tumour necrosis factor-alpha (TNF-alpha), alanine amino transferase (ALT), and HCV viral load. Study design: The susceptibility of peripheral blood mononuclear cells from 17 subjects with chronic HCV infection to Fas ligand induced cell death was assessed using a water soluble tetrazolium assay. The plasma levels of associated markers such as sFas, sFasL, and TNF-alpha were quantified using immunoassays. ALT values were obtained from hospital records. Viral loads were quantified using a commercially available quantitative assay - the Amplicor Monitor (version 2.0). Controls for comparison included a group of healthy individuals and individuals infected with the human immunodeficiency virus 1. Results: The percentage of cell death induced in hepatitis C virus infected individuals was lower than that seen in the healthy control group. Patients infected with HCV had higher average values of sFas and TNF-alpha as compared to both control groups. Plasma levels of sFas in patients with chronic HCV infection showed significant positive correlations to ALT and TNF-alpha levels. TNF-alpha levels also showed a significant positive correlation with ALT levels. Conclusions: PBMC in HCV infection exhibit decreased susceptibility to Fas ligand induced cell death. This may signify a means by which HCV escapes immune surveillance. This phenomenon merits further investigation. The strong correlations observed between plasma sFas, ALT and TNF-alpha suggest a potential role for these markers as an alternative to an invasive liver biopsy

Evaluation of a Rapid Assay as an Alternative to Conventional Enzyme Immunoassays for Detection of Hepatitis C Virus-Specific Antibodies

A rapid membrane flow-through immunoassay to detect antibodies to hepatitis C virus was compared with a commercial enzyme immunoassay (EIA) and microparticle enzyme immunoassay (MEIA) using 2,590 serum samples. Sensitivity and specificity of the “rapid assay” in comparison to the EIA/MEIA were 99.3 and 99.0%; the correlation coefficient being 0.91. This assay is suitable where infrastructure and laboratory expertise are limited

HCV genotype 4 - an emerging threat as a cause of chronic liver disease in Indian (South) patients

Background: Hepatitis C virus (HCV) genotyping is relevant for the delivery of effective antiviral therapy. HCV genotypes are geographically restricted with genotype 4, which is resistant to therapy, traditionally considered to be confined to the Middle East and Africa. We report here on the occurrence of HCV genotype 4 in Indian (South) patients. Objectives: (1)To highlight the occurrence of HCV genotype 4 in the patient population attending a tertiary care hospital in south India. (2)To ascertain the difference in HCV viral loads and alanine aminotransferase (ALT) values between patients infected with HCV genotype 4 and those infected with the other two most commonly detected genotypes in this patient population viz., HCV genotypes 1 and 3. (3)To assess the genetic relatedness of the Indian strains to Genbank sequences, which we report for the first time. Study design: The study group consisted of 125 HCV infected, untreated patients who had been genotyped using type specific primers. Eight of the nine samples classified as HCV genotype 4 by this technique were subjected to nucleotide sequencing. Viral load estimations were carried out. Information on possible risk factors and ALT values were obtained from hospital records. Statistical analyses were carried out to compare viral loads and ALT values across genotypes. A phylogenetic tree was constructed and the genetic relatedness of the strains was assessed through sequence analysis. Results: HCV genotype 4 was detected in nine of 125 (7.2%) patients. Eight of the nine were subjected to nucleotide sequencing and all strains were confirmed as HCV genotype 4. Six of the eight strains were closely related, with two strains being phylogenetically diverse. Conclusions: HCV genotype 4 is detected in a significant minority of HCV infected patients in India. This finding should be considered in designing strategies prior to initiation of therapy in Indian patients infected with HCV