Hydrogen Peroxide Preconditioning Promotes Protective Effects of Umbilical Cord Vein Mesenchymal Stem Cells in Experimental Pulmonary Fibrosis

Purpose Idiopathic pulmonary fibrosis (IPF) is a progressive lung disorder with few available treatments. Mesenchymal stem cell therapy (MSCT), an innovative approach, has high therapeutic potential when used to treat IPF. According to recent data, preconditioning of MSCs can improve their therapeutic effects. Our research focuses on investigating the anti-inflammatory and antifibrotic effects of H2O2-preconditioned MSCs (p-MSCs) on mice with bleomycin-induced pulmonary fibrosis (PF). Methods Eight-week-old male C57BL/6 mice were induced with PF by intratracheal (IT) instillation of bleomycin (4 U/kg). Human umbilical cord vein-derived MSCs (hUCV-MSCs) were isolated and exposed to a sub-lethal concentration (15 pM for 24 h) of H2O2 in vitro. One week following the injection of bleomycin, MSCs or p-MSCs were injected (IT) into the experimental PF. The survival rate and weight of mice were recorded, and 14 days after MSCs injection, all mice were sacrificed. Lung tissue was removed from these mice to examine the myeloperoxidase (MPO) activity, histopathological changes (hematoxylin-eosin and Masson\u27s trichrome) and expression of transforming growth factor beta 1 (TGF-β1) and alpha-smooth muscle actin (α-SMA) through immunohistochemistry (IHC) staining. Results Compared to the PF+MSC group, p-MSCs transplantation results in significantly decreased connective tissue () and collagen deposition. Additionally, it is determined that lung tissue in the PF+pMSC group has increased alveolar space () and diminished expression of TGF-β1 and α-SMA. Conclusion The results demonstrate that MSCT using p-MSCs decreases inflammatory and fibrotic factors in bleomycin-induced PF, while also able to increase the therapeutic potency of MSCT in IPF

Evaluation of LOXL1 gene polymorphisms in exfoliation syndrome and exfoliation glaucoma

Purpose: To evaluate genetic susceptibility of lysyl oxidase-like 1 (LOXL1) gene polymorphisms to exfoliation syndrome (XFS) and exfoliation glaucoma (XFG) in a case-control cohort of American and European patients. Methods: DNA from a total of 620 individuals including 287 exfoliation patients and 333 healthy control subjects were extracted by standard methods. Three single nucleotide polymorphisms (SNPs) of rs1048661 (R141L), rs3825942 (G153D), and rs2165241 were genotyped in these individuals by SNaPshot Assay. The seven coding exons of the LOXL1 gene and their immediate flanking regions were directly sequenced in 95 affected patients. Data management and case-control association studies were performed with SNP-STAT and PLINK programs. The obtained DNA sequences were evaluated with the STADEN package. Results: The 287 unrelated exfoliation cases comprised of 171 American patients (mostly of European background) and 116 patients from 12 European countries. This phenotype was further divided into patients with exfoliation only and no glaucoma (XFO; n=95), exfoliation with glaucoma (XFG; n=133), and exfoliation unclassified (XFU; n=59). Genotypic data were analyzed separately for XFO, XFG, XFU, and XFS (all exfoliations; n=287) and for Americans and Europeans. The observed genotypic frequencies for each exfoliation phenotype or population were tabulated separately and tested for deviation from the Hardy–Weinberg equilibrium (HWE) using a standard Χ2 test. There were no HWE deviations and no significant genotypic differences between these subcategories for the three studied SNPs. For the combined exfoliation cohort, homozygote genotypes of G/G (rs1048661), G/G (rs3825942), and T/T (rs2165241) were significantly overrepresented. Likewise, case-control allelic association for rs1048661 (p=7.74x10−9), rs3825942 (p=3.10x10−17), and rs2165241 (p=4.85x10−24) were highly significant. The corresponding two-locus haplotype frequencies of GG for rs1048661-rs3825942 (p=1.47x10−27), GT for rs1048661-rs2165241 (p=1.29x10−24), and GT for rs3825942-rs2165241 (p=2.02x10−24) were highly associated with exfoliation phenotypes. The combined effect of these three SNPs revealed that the GGT haplotype is overrepresented by 66% in exfoliation cases, and this deviation from controls is highly significant (p=1.93x10−24). This haplotype constituted a major risk factor for development of exfoliation in both XFS and XFG. By contrast, the GAC haplotype was significantly underrepresented (p=4.99x10−18) in exfoliation cases by 83% and may potentially have a protective effect for this condition with an estimated attributable risk percent reduction of 457%. The only other haplotype that was significantly different between cases and controls was TGC (p=5.82x10−9). No observation was made for the GAT haplotype. The combined three haplotypes of GGT, GAC, and TGC were associated with 91% of the exfoliation syndrome cases in the studied populations. Seven coding exons of LOXL1 were also sequenced in 95 affected cases. In addition to the three above-mentioned SNPs, 12 other variations were also observed in these patients(G240G, D292D, A320A, V385V, rs2304719, IVS3+23C>T, IVS3–155G>A, IVS3–101G>A, IVS4+49G>A, rs2304721, IVS5–121C>T, and rs2304722). None were considered a disease-causing mutation. Conclusions: We confirmed a strong association with LOXL1 variants in our patients. For the LOXL1 gene, individual alleles of rs1048661 (G), rs3825942 (G), and rs2165241 (T) are highly associated with XFS and XFG in American and European populations. The GGT haplotype constitutes a major risk haplotype for exfoliation, and GAC may have a protective role. DNA sequencing of 95 affected patients did not show any mutations in this gene. The LOXL1 SNPs are located in the 15q24.1 band and within a genetic locus (GLC1N) that is associated with primary open-angle glaucoma (POAG). However, the LOXL1 genetic predisposition is only limited to exfoliation with or without glaucoma and does not include the POAG phenotype

Molecular determination of low tension primary open angle glaucoma gene (GLCIE on 10p15-10p14)

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Determination of sensitivity, specificity and cut off point of visual- Motor Bender Gestalt Test in the diagnosis of traumatic brain injury

Background: Bender Gestalt Test is one of the most famous neuropsychological tests, simple and easy to perform, and is used to evaluate brain injuries. This study aimed at determining the rate of sensitivity, characteristic and cut-off point of this test in patients with traumatic brain injury (TBI). Methods and Materials: Overall, 120 TBI patients with mean age of 31.25± 13.60 years old in a descriptive-analytical research design entered the study using nonprobability and consecutive sampling method. All patients underwent Bender Visual-Motor Gestalt Test after neurological evaluations by CT scan. Roc curve test was utilized to analyze the data. Results: In this study, cut-off point was calculated as 6.5%, sensitivity as 55.8%, characteristic as 81.2%, and the area under the Roc curve as 0.69. Moreover, positive predictive value, negative predictive value and efficiency were 95.08%, 22.03%, and 59.17%, respectively. Conclusion: Results of this study revealed that Bender Gestalt Test is relatively weak in diagnosis of mild TBI. Hence, its characteristic is high and it was successful in diagnosing healthy individuals