206 research outputs found

    Improved Differentiation of Mesenchymal Stem Cells into Hepatocyte-like Cells using FGF4 and IGF-1 in 3D Culture

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    Human Umbilical Cord Mesenchymal Stem Cells (UCMSCs) are considered as an excellent candidate for cell therapy to treat end-stage liver disease. Fibroblast Growth Factor-4 (FGF4), Hepatocyte Growth Factor, and Insulin-like Growth Factor-1 are some of the critical cytokines involved in liver development and regeneration. To evaluate the differentiation potency of cells into hepatocyte-like cells we used these cytokines. UCMSCs were isolated from Wharton's jelly of fullterm infants. The cells were characterized as MSCs by flow-cytometry and their multilineage differentiation capacity. Then, UCMSCs were cultured in 3D collagen scaffold and hepatogenic media with or without FGF4 for 21 days and the data were compared to control. The expression of liver specific genes was evaluated by real-time quantitative RT-PCR and immunocytochemistry. These cells expressed MSC markers and could differentiate into adipocytes and osteocytes. A non–significant higher level of liver specific genes, such as cytokeratin-18 and 19, alpha-fetoprotein and albumin, and also a significant higher level of CYP2B6 expressed by UCMSCs in hepatogenic medium containing FGF4 compared with control. In some specimens, cytokeratin-19-positive cells surrounded a luminal space within collagen scaffolds. Liver-specific marker expression was increased by pre-exposing the cells to FGF4 before treating with IGF-1 and HGF in 3D collagen scaffold. Abbreviations: UCMSCs: Human Umbilical Cord Mesenchymal Stem Cells; FGF4: Fibroblast Growth Factor 4; HGF: Hepatocyte Growth Factor; IGF-1: Insulin-like Growth Factor-1; MSCs: Mesenchymal Stem Cells; ICG: Indocyanine green; PAS: periodic acid Schiff; CK-18: cytokeratin-18; CK-19: Cytokeratin-19; AFP: alpha-fetoprotein; G6P: glucose 6 phosphatase; PEPCK: phosphoenolpyruvate carboxykinase; TAT: tyrosine amino transferase; FBS: Fetal Bovine Serum; OSM: oncostatin M; RT-PCR: Reverse Transcription Polymerase Chain Reaction; PBS: Phosphate-Buffered Saline; Hep- Par1: Hepatocyte paraffin 1; DAB: Diaminobenzidine; CYP2B6: Cytochrome P450 2B6

    Polymorphism at High Molecular Weight Glutenin Subunits and Morphological Diversity of Aegilops geniculata Roth Collected in Algeria

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    A collection of 35 accessions of the tetraploid wild wheat Aegilops geniculata Roth (MM, UU) sampled in northern Algeria was evaluated for morphological and biochemical variability. Morphological and ecological analyses based on morphological traits and bioclimatic parameters, respectively, were assessed using principal component analysis (PCA). Accessions were differentiated by width characters, namely spike’s width, and a weak relationship between morphological traits and ecological parameters was found. Polymorphism of high molecular weight (HMW) glutenin subunits was carried on by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Among accessions analyzed, 27 alleles were identified at the two loci Glu-M1 and Glu-U1: resulting in twenty-nine patterns and a nomenclature was proposed. Two alleles at the Glu-U1 locus expressed a new subunit with a slightly slower mobility than subunit 8. These results provide new information regarding the genetic variability of HMW glutenin subunits, as well as their usefulness in cultivated wheat quality improvement

    Novel metabolic biomarkers related to sulfur-dependent detoxification pathways in autistic patients of Saudi Arabia

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    <p>Abstract</p> <p>Background</p> <p>Xenobiotics are neurotoxins that dramatically alter the health of the child. In addition, an inefficient detoxification system leads to oxidative stress, gut dysbiosis, and immune dysfunction. The consensus among physicians who treat autism with a biomedical approach is that those on the spectrum are burdened with oxidative stress and immune problems. In a trial to understand the role of detoxification in the etiology of autism, selected parameters related to sulfur-dependent detoxification mechanisms in plasma of autistic children from Saudi Arabia will be investigated compared to control subjects.</p> <p>Methods</p> <p>20 males autistic children aged 3-15 years and 20 age and gender matching healthy children as control group were included in this study. Levels of reduced glutathione (GSH), total (GSH+GSSG), glutathione status (GSH/GSSG), glutathione reductase (GR), glutathione- s-transferase (GST), thioredoxin (Trx), thioredoxin reductase (TrxR) and peroxidoxins (Prxs I and III) were determined.</p> <p>Results</p> <p>Reduced glutathione, total glutathione, GSH/GSSG and activity levels of GST were significantly lower, GR shows non-significant differences, while, Trx, TrxR and both Prx I and III recorded a remarkably higher values in autistics compared to control subjects.</p> <p>Conclusion</p> <p>The impaired glutathione status together with the elevated Trx and TrxR and the remarkable over expression of both Prx I and Prx III, could be used as diagnostic biomarkers of autism.</p

    Improvement of attention span and reaction time with hyperbaric oxygen treatment in patients with toxic injury due to mold exposure

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    It is, by now, well established that mold toxins (mycotoxins) can cause significant adverse health effects. In this study, 15 subjects who developed an attention deficit disorder (ADD) and slowing of reaction time at the time of exposure to mold toxins were identified. Deficits in attention span and reaction time were documented not only by taking a careful history, but also by performing a Test of Variables of Attention (TOVA). The TOVA test provides an objective measure of these two variables. It was found that mold-exposed subjects show statistically significant decreases in attention span and significant increases in reaction time to stimuli compared to controls. After ten sessions of hyperbaric oxygen treatment (HBOT), a statistically significant improvement was seen in both measures. This preliminary study suggests promising outcomes in treating mold-exposed patients with hyperbaric oxygen

    In Search of Cellular Immunophenotypes in the Blood of Children with Autism

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    Autism is a neurodevelopmental disorder characterized by impairments in social behavior, communication difficulties and the occurrence of repetitive or stereotyped behaviors. There has been substantial evidence for dysregulation of the immune system in autism.We evaluated differences in the number and phenotype of circulating blood cells in young children with autism (n = 70) compared with age-matched controls (n = 35). Children with a confirmed diagnosis of autism (4-6 years of age) were further subdivided into low (IQ<68, n = 35) or high functioning (IQ ≥ 68, n = 35) groups. Age- and gender-matched typically developing children constituted the control group. Six hundred and forty four primary and secondary variables, including cell counts and the abundance of cell surface antigens, were assessed using microvolume laser scanning cytometry.There were multiple differences in immune cell populations between the autism and control groups. The absolute number of B cells per volume of blood was over 20% higher for children with autism and the absolute number of NK cells was about 40% higher. Neither of these variables showed significant difference between the low and high functioning autism groups. While the absolute number of T cells was not different across groups, a number of cellular activation markers, including HLA-DR and CD26 on T cells, and CD38 on B cells, were significantly higher in the autism group compared to controls.These results support previous findings that immune dysfunction may occur in some children with autism. Further evaluation of the nature of the dysfunction and how it may play a role in the etiology of autism or in facets of autism neuropathology and/or behavior are needed

    Complexity of the Mycoplasma fermentans M64 Genome and Metabolic Essentiality and Diversity among Mycoplasmas

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    Recently, the genomes of two Mycoplasma fermentans strains, namely M64 and JER, have been completely sequenced. Gross comparison indicated that the genome of M64 is significantly bigger than the other strain and the difference is mainly contributed by the repetitive sequences including seven families of simple and complex transposable elements ranging from 973 to 23,778 bps. Analysis of these repeats resulted in the identification of a new distinct family of Integrative Conjugal Elements of M. fermentans, designated as ICEF-III. Using the concept of “reaction connectivity”, the metabolic capabilities in M. fermentans manifested by the complete and partial connected biomodules were revealed. A comparison of the reported M. pulmonis, M. arthritidis, M. genitalium, B. subtilis, and E. coli essential genes and the genes predicted from the M64 genome indicated that more than 73% of the Mycoplasmas essential genes are preserved in M. fermentans. Further examination of the highly and partly connected reactions by a novel combinatorial phylogenetic tree, metabolic network, and essential gene analysis indicated that some of the pathways (e.g. purine and pyrimidine metabolisms) with partial connected reactions may be important for the conversions of intermediate metabolites. Taken together, in light of systems and network analyses, the diversity among the Mycoplasma species was manifested on the variations of their limited metabolic abilities during evolution
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