Twenty-seven mutations with three novel pathologenic variants causing biotinidase deficiency: a report of 203 patients from the southeastern part of Turkey

BACKGROUND: Biotinidase deficiency (BD) is an autosomal recessive inborn error of metabolism characterized by neurologic and cutaneous symptoms and can be detected by newborn screening. Newborn screening for BD was implemented in Turkey at the end of 2008. METHODS: In total, 203 patients who were identified among the infants detected by the newborn screening were later confirmed to have BD through measurement of serum biotinidase activity. We also performed BTD mutation analysis to characterize the genetic profile. RESULTS: Twenty-seven mutations were identified. The most commonly found variants were c.1330G>C (p.D444H), c.1595C>T (p.T532M), c.470G>A (p.R157H), and c.198_104delGCGGCTGinsTCC (p.C33Ffs ) with allele frequencies of 0.387, 0.175, 0.165 and 0.049, respectively. Three novel pathogenic and likely pathogenic variants were identified: p.W140* (c.419G>A), p.S319F (c.956C>T) and p.L69Hfs*24 (c.192_193insCATC). We also identified three mutations reported in just one patient in the past (p.V442Sfs*59 [c.1324delG], p.H447R [c.1340A>G] and p.198delV [c.592_594delGTC]). Although all of the patients were asymptomatic under the treatment of biotin, only one patient, who had the novel c.419G>A homozygous mutation became symptomatic during an episode of acute gastroenteritis with a presentation of ketosis and metabolic acidosis. Among the screened patients, 156 had partial and 47 had profound BD. CONCLUSIONS: We determined the mutation spectra of BD from the southeastern part of Turkey. The results of this study add three more mutations to the total number of mutations described as causing BD

Amphipathic Cell-Penetrating Peptide-Aided Delivery of Cas9 RNP for In Vitro Gene Editing and Correction

The therapeutic potential of the CRISPR-Cas9 gene editing system in treating numerous genetic disorders is immense. To fully realize this potential, it is crucial to achieve safe and efficient delivery of CRISPR-Cas9 components into the nuclei of target cells. In this study, we investigated the applicability of the amphipathic cell-penetrating peptide LAH5, previously employed for DNA delivery, in the intracellular delivery of spCas9:sgRNA ribonucleoprotein (RNP) and the RNP/single-stranded homology-directed repair (HDR) template. Our findings reveal that the LAH5 peptide effectively formed nanocomplexes with both RNP and RNP/HDR cargo, and these nanocomplexes demonstrated successful cellular uptake and cargo delivery. The loading of all RNP/HDR components into LAH5 nanocomplexes was confirmed using an electrophoretic mobility shift assay. Functional screening of various ratios of peptide/RNP nanocomplexes was performed on fluorescent reporter cell lines to assess gene editing and HDR-mediated gene correction. Moreover, targeted gene editing of the CCR5 gene was successfully demonstrated across diverse cell lines. This LAH5-based delivery strategy represents a significant advancement toward the development of therapeutic delivery systems for CRISPR-Cas-based genetic engineering in in vitro and ex vivo applications

The role of the family in attributing meaning to living with HIV and its stigma in Turkey

Stigma attached to HIV/AIDS remains a global problem, with severe negative consequences for people living with HIV (PLHIV). Family support is fundamental for PLHIV’s psychological and physical well-being. HIV-related stigma is high in Turkey, where HIV/AIDS prevalence is low and the epidemic is not considered a priority. Based on qualitative data generated with HIV-positive women and men, this article explores the process of stigmatization, as experienced and perceived by PLHIV in Turkey, focusing on the institution of the family. Results indicated that enacted stigma from family members is lower than anticipated. While most participants’ narratives showed patterns of support rather than rejection from families, the strong expectations around the cultural value attributed to “the family” are found to be the main facilitators of internalized stigma. The article critically discusses the meaning and implications of family support, addressing the role of patriarchal values attributed to womanhood, manhood, and sexuality in Turkey

Functionalization of low molecular weight atactic polypropylene Part II. Thermal and molecular weight studies

Number average molecular weight (M(n)), intrinsic viscosity ([?]) and glass transition temperature (T(g)) of low molecular weight atactic polypropylene (APP) grafted with 4-allyl-1,2-dimethoxybenzene (DMAB), 4-allyl-2-methoxyphenol (eugenol) and 4-propenylanisole (trans-anethole) in the presence of dicumyl peroxide (DCP) at 170°C were determined. The influences of reaction time, concentration and extent of grafting on M(n), [?] and T(g) of APP were examined. The data were discussed with the mechanism of grafting reactions

Novel hydrophobic ligand-containing hydrogel membrane matrix: Preparation and application to ?-globulins adsorption

In this study, phenylalanine as a hydrophobic ligand was covalently attached to the co-monomer methacrylochloride. Then, poly(2-hydroxyethylmethacrylate-co-methacrylamidophenyalanine) [poly(HEMA-MAPA)] membranes were prepared by UV-initiated photopolymerization of HEMA and methacrylamidophenyalanine. The ?-globulins adsorption onto these affinity membranes from aqueous solutions containing different amounts of ?-globulins at different pH was investigated in a batch system. The ?-globulins adsorption capacity of the membranes was increased as the ligand density on the membrane surface increase. The non-specific adsorption of the ?-globulins on the pHEMA membranes was negligible. The adsorption phenomena appeared to follow a typical Langmuir isotherm. The maximum adsorption capacity (qm) of the poly(HEMA-MAPA4) membrane for ?-globulins was 2.37 mg g-1dry membrane. The equilibrium constant (kd) value was found to be 1.61×10-1mg ml-1. More than 87% (up to 100%) of the adsorbed ?-globulins were desorbed in 120 min in the desorption medium containing 50% ethylene glycol in 1.0 M NaCl. Copyright © 2001 Elsevier Science B.V

The impact of granulocyte colony-stimulating factor (G-CSF) on thin endometrium of an animal model with rats

Purpose: To evaluate whether G-CSF improves the endometrial thickness of thin endometrium by influencing proliferative, angiogenic and apoptotic factors, an experimental rat model was conducted using 24 female adult rats with either thin or healthy endometrium that each was further divided into G-CSF or saline injection groups with six rats. Materials and methods: After forming of the thin endometrium by uterine injection of 0.2 ml 96% ethyl alcohol to the rats, five days of subcutaneous injections of 40 μg/kg G-CSF or saline were given. Endometrial thickness, immunohistochemically expression of vascular endothelial growth factor receptor-2 (VEGF-R2), proliferative cell nuclear antigen (PCNA) and fibronectin apoptosis with TUNEL method were compared in specimens among four groups of post-model rats. Results: Endometrial thickness was significantly improved in thin but not in normal endometrium group with GCSF when compared to saline injection. Stromal and glandular epithelial expression of PCNA and pericapillary VEGF-R2 was significantly increased, and apoptosis was significantly decreased with G-CSF. Although fibronectin was also increased with G-CSF in the thin endometrium, the difference was non-significant. In further, G-CSF decreased apoptotic cells and increased expression of PCNA when compared to saline injection in normal endometrium. Conclusions: G-CSF improves endometrial thickness, proliferation, angiogenesis and DNA fragmentation in thin endometrium. © 2020 Informa UK Limited, trading as Taylor & Francis Group

Thermal properties and flame retardance of Al(OH)3/polypropylene composites modified by polypropylene grafting with acrylic acid

The effect of polypropylene grafting with acrylic acid, PP-g-AA (FPP), on crystallization and melting behavior, thermal degradation, and limiting oxygen index of Al(OH)3/PP composites were investigated. The results indicated that crystallization temperature of PP shifted to high temperature with increasing content of Al(OH)3 because of the interfacial heterogeneous nucleation of Al(OH)3 and further increased by the addition of FPP and with increasing FPP content because of the improvement of the dispersion of Al(OH)3 in PP matrix and the increase in the nucleating sites of Al(OH)3. With adding Al(OH)3 and increasing the content of Al(OH)3, limiting oxygen index values of composites increased and further improved by adding FPP. This is attributed to the presence of an interfacial interaction between FPP and Al(OH)3. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci