Biallelic KITLG variants lead to a distinct spectrum of hypomelanosis and sensorineural hearing loss.

BACKGROUND: Pathogenic variants in KITLG, a crucial protein involved in pigmentation and neural crest cell migration, cause non-syndromic hearing loss, Waardenburg syndrome type 2, familial progressive hyperpigmentation and familial progressive hyper- and hypopigmentation, all of which are inherited in an autosomal dominant manner. OBJECTIVES: To describe the genotypic and clinical spectrum of biallelic KITLG-variants. METHODS: We used a genotype-first approach through the GeneMatcher data sharing platform to collect individuals with biallelic KITLG variants and reviewed the literature for overlapping reports. RESULTS: We describe the first case series with biallelic KITLG variants; we expand the known hypomelanosis spectrum to include a 'sock-and-glove-like', symmetric distribution, progressive repigmentation and generalized hypomelanosis. We speculate that KITLG biallelic loss-of-function variants cause generalized hypomelanosis, whilst variants with residual function lead to a variable auditory-pigmentary disorder mostly reminiscent of Waardenburg syndrome type 2 or piebaldism. CONCLUSIONS: We provide consolidating evidence that biallelic KITLG variants cause a distinct auditory-pigmentary disorder. We evidence a significant clinical variability, similar to the one previously observed in KIT-related piebaldism.

Evaluation of the NOX5 protein expression and oxidative stress in sperm from asthenozoospermic men compared to normozoospermic men.

PURPOSE: NADPH oxidase 5 (NOX5), the main isoform of NOX in spermatozoa, has been recognized as the main active generators of reactive oxygen species (ROS), including superoxide anion (O 2 -. ) and hydrogen peroxide (H2O2). ROS have been shown to play important roles in many physiological and pathological conditions in spermatozoa. The present study aims to investigate the alterations of NOX5 protein expression and oxidative stress (OS) status in asthenozoospermic men compared to normozoospermic men. METHODS: Semen samples were collected from 25 asthenozoospermic men and 28 normozoospermic men. In this study, NOX5 protein expression was evaluated by Western blotting. An OS status was evaluated by measuring of ROS (O 2 -. and H2O2), DNA damage and plasma membrane integrity in spermatozoa. RESULTS: The protein expression of NOX5 (p < 0.0001) was remarkably higher in asthenozoospermic men in comparison to normozoospermic men. In addition, the percentages of intracellular O 2 -. (p < 0.0001), H2O2 (p < 0.0001) in viable spermatozoa, apoptotic sperm cells with altered plasma membrane (p < 0.001) and DNA damage (p = 0.001) were significantly increased in asthenozoospermic men compared to normozoospermic men. CONCLUSIONS: The present study provides evidence that the overexpression of NOX5 protein may induce excessive ROS production and oxidative stress damages to DNA and plasma membrane integrity in asthenozoospermic men.

Circulating Levels of LAMP2 in Coronary Artery Disease: Association with Serum Lipid Profile.

Whereas several in vitro and in vivo studies have described the role of lysosomal associated membrane protein 2 (LAMP2) in lipid homeostasis, there is no study addressing LAMP2 serum concentration and its association with lipid profiles in the context of coronary artery disease (CAD). We aimed to determine the LAMP2 serum concentration and its association with serum lipid profiles as well as the gene expression of LAMP2 in peripheral blood mononuclear cells (PBMCs) of CAD patients and control group. Circulating levels of LAMP2 were quantified by enzyme-linked immunosorbent assay (ELISA) in CAD patients (n=85) and control group (n=65) and correlation to lipid parameters was assessed. Gene expression analysis was performed by quantitative real-time PCR. Mean LAMP2 serum concentration adjusted for drug consumption, age and gender was not significantly different between the CAD and control groups (p>0.05). However, LAMP2 serum concentration showed independent significant association with lipid profiles including triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) (all p<0.05). Furthermore, increased expression of LAMP2 has been observed in PBMCs of CAD patients compared to the control group (p<0.05). Our findings supported the previous observations showing the contribution of LAMP2 in lipid homeostasis and pathogenesis of CAD.

The mRNA Expression and Circulating Levels of Visfatin and Their Correlation with Coronary Artery Disease Severity and 25-Hydroxyvitamin D.

It is evident that coronary artery disease (CAD) is closely associated with abnormal glucose and lipid metabolism. Notably, dysregulation of inflammatory pathways and immune system also contribute to CAD development. Recently, it has been suggested that visfatin, a proinflammatory adipocytokine, may be involved in several inflammatory and metabolic diseases. In this study, we evaluated the serum visfatin levels and its mRNA expression in peripheral blood mononuclear cells (PBMCs) from CAD patients compared with control subjects. We also studied the correlation between visfatin gene expression and serum levels with clinical and metabolic parameters. This study was conducted on 56 male patients with CAD confirmed by angiography and 30 healthy men as controls. CAD severity was determined based on the number of vessels. Study of gene expression in PBMCs was performed using real time-PCR, and serum levels of visfatin and 25-hydroxyvitamin D were measured by ELISA. We found that serum visfatin levels and its gene expression in PBMCs were increased in patients with CAD compared with the control group (p=0.027 and p=0.016, respectively). Also, visfatin gene expression was positively correlated with visfatin levels and both these variables had a strong positive correlation with the severity of CAD. It appears that elevated mRNA expression and circulating level of visfatin might be of relevance to the pathogenesis and severity of CAD. However, further studies are necessary to better clarify the associations between visfatin and CAD.

CTRP1 in Liver Disease.

Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease occurs in significant percentage of general population. NAFLD is closely associated with entire spectrum of metabolic-related disorders including diabetes, obesity, and cardiovascular diseases. Considering several similar pathways underpinning metabolic disorders, presence of common molecular mediators contributing to pathomechanism of these disorders is expected. Mounting evidence has demonstrated important role of adipokines in the context of NAFLD. Adipokines produced by different tissues, mainly adipose, modulate numerous pathways including glucose and fatty acid metabolism and inflammation. CTRPs (C1q/TNF-related proteins) are a recently identified family of adipokines in which adiponectin is the most well-known ones. CTRP1 is a member of this family which has captured attention in recent years. CTRP1 enhances glucose and fatty acid oxidation, improves insulin sensitivity, attenuates plaque formation, and increases aldosterone production. Hence, various roles in metabolic pathways can link CTRP1 to NAFLD pathogenesis.

Prenatal maternal immune activation increases anxiety- and depressive-like behaviors in offspring with experimental autoimmune encephalomyelitis.

Multiple sclerosis (MS) is thought to result from a combination of genetics and environmental factors. Several lines of evidence indicate that significant prevalence of anxiety and depression-related disorders in MS patients can influence the progression of the disease. Although we and others have already reported the consequences of prenatal maternal immune activation on anxiety and depression, less is known about the interplay between maternal inflammation, MS and gender. We here investigated the effects of maternal immune activation with Poly I:C during mid-gestation on the progression of clinical symptoms of experimental autoimmune encephalomyelitis (EAE; a mouse model of MS), and then anxiety- and depressive-like behaviors in non-EAE and EAE-induced offspring were evaluated. Stress-induced corticosterone and tumor necrosis factor-alpha (TNF-α) levels in EAE-induced offspring were also measured. Maternal immune activation increased anxiety and depression in male offspring, but not in females. This immune challenge also resulted in an earlier onset of the EAE clinical signs in male offspring and enhanced the severity of the disease in both male and female offspring. Interestingly, the severity of the disease was associated with increased anxiety/depressive-like behaviors and elevated corticosterone or TNF-α levels in both sexes. Overall, these data suggest that maternal immune activation with Poly I:C during mid-pregnancy increases anxiety- and depressive-like behaviors, and the clinical symptoms of EAE in a sex-dependent manner in non-EAE or EAE-induced offspring. Finally, the progression of EAE in offspring seems to be linked to maternal immune activation-induced dysregulation in neuro-immune-endocrine system.

Serological Study on Cytomegalovirus and Toxoplasma Gondii in Thalassemia Major Patients of Yazd, Iran.

BACKGROUND: Beta-thalassemia patients receive blood products from blood transfusion centers repeatedly. Blood transfusion can transmit Cytomegalovirus (CMV) and Toxoplasma gondii. The aim of this study was serological evaluation of these two infectious agents in thalassemia patients. MATERIALS AND METHODS: In a cross-sectional study, the enzyme-linked immunosorbent assay (ELISA) testing was performed to detect IgM and IgG antibodies against CMV and Toxoplasma gondii in 96 thalassemia patients (under 18 years) and 144 healthy people. Data were analyzed by SPSS software and Chi-square test. RESULTS: A significant difference was observed in CMVIgM antibody levels between test groups in women (p<0.05). The prevalence of CMV IgM, CMV IgG, Toxo-IgG, and Toxo IgM antibodies in thalassemia patients were 5.2%, 95.9%, 16%, and 0%, respectively. CONCLUSION: In all thalassemia patients, Cytomegalovirus IgG is higher than healthy people. In addition, CMV IgM antibodies are higher in female patients. Antibody screening (IgM) on blood products for detecting Cytomegalovirus is necessary, but for Toxoplasma gondii is not necessary in the Yazd transfusion center.

Effect of in ovo injection of threonine on Mucin2 gene expression and digestive enzyme activity in Japanese quail (Coturnix japonica).

A total of 540 Japanese quail eggs were assigned to 9 treatments of 4 replicates to investigate the effect of in ovo injection of threonine (THR) on mucin2 (MUC2) mRNA expression and digestive enzyme activity. Treatments were (non-injected) eggs and those in ovo injected with saline (0.05 or 0.1 ml) with or without THR (5 mg/ml) in two sites (in or under the air sac). On hatch day, 0.05 ml in ovo injected (under the air sac: TUAS) hatchlings were divided into three groups based on NRC recommendations for THR, while all 0.1 ml in ovo injected chicks were removed due to low hatchability. The remaining treatments received the NRC recommended diet until day 10 post-hatch. Treatments had no effect on protease and amylase activities, while TUAS increased MUC2 gene expression. In conclusion, the in ovo injection of THR increased MUC2 gene expression but had no effect on enzyme activity.

Apolipoprotein B as the best predictor of coronary artery disease in Iranian normolipidemic patients.

OBJECTIVES: A relatively high proportion of Iranian patients with coronary artery disease (CAD) have normal levels of traditional lipid risk factors and show early onset of CAD. In this study we examined the roles of apolipoprotein B (apoB), apolipoprotein AI (apoAI) and lipoprotein (a) [LP(a)] in predicting coronary heart disease in normolipidemic patients and those with premature CAD (age < or = 50). DESIGN AND METHODS: Serum levels of apoB, apoAI, and LP(a) were determined in a total of 567 Iranian patients who were candidates for coronary angiography. A subgroup of 142 patients (93 males, 49 females) with normal levels of classical lipid risk factors, and a subgroup of patients (130 males, 71 females) with age below 50 years were separately assessed for coronary risk factors. RESULTS: ApoB concentrations were significantly higher in patients with CAD (CAD+) relative to patients without CAD (CAD-) in the two subgroups. Multiple logistic regression after controlling for age and others risk factors showed apoB as the best determinant of CAD in the normolipidemic subgroup (OR, 4.3, p < 0.001) and in the men aged < or = 50 (OR, 5.7, p < 0.001). ApoB was the best predictor of CAD in a subgroup of very young patients (age < or = 40, n = 77, OR, 8.6, p < 0.009). There was a significant correlation between severity of atherosclerosis and serum apoB concentration in the normolipidemic subgroup (r = 0.22, p < 0.008). CONCLUSIONS: Our data indicate that serum concentration of apoB is the best discriminating factor to predict the presence or absence of atherosclerosis in Iranian normolipidemic individuals and young patients undergoing coronary angiography.

Preparative SDS-PAGE Electroelution for Rapid Purification of Alkyl Hydroperoxide Reductase from Helicobacter pylori.

BACKGROUND: Alkyl hydroperoxide reductase (AhpC) of Helicobacter pylori is considered as a diagnostic antigen. Therefore, this antigen can be used to detect H. pylori infection by stool immunoassays such as ELISA. The aim of this study was to simplify the AhpC protein purification procedures. METHODS: For whole cell protein extraction, the bacterial cells were ruptured by octly-ß-D glucopyranoside. The isolation and purification of AhpC protein were attempted by various techniques including ammonium sulfate precipitation, dialysis, preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electroelution. RESULTS: A simple method was used for protein purification AhpC protein. One-dimensional preparative gel electrophoresis allows a single and short purification step; the high resolution capacity of this technique leads to a high level of purity of the protein. Moreover, it avoids contamination by other non-specific proteins which often appear during protein purification by column chromatography. CONCLUSION: The present method is simple, rapid and makes it possible to preparate AhpC from H. pylori.

Decreased polyunsaturated and increased saturated fatty acid concentration in spermatozoa from asthenozoospermic males as compared with normozoospermic males.

The lipid composition of the sperm membrane has been shown to exert a significant effect upon the functional quality of spermatozoa. We have studied fatty acid composition of the phospholipids in spermatozoa in asthenozoospermic and normozoospermic men and determined the ratio of polyunsaturated fatty acids (PUFAs) to saturated fatty acids of spermatozoa of these two groups. Fatty acid concentration of spermatozoa was determined in 15 asthenozoospermic and eight normozoospermic semen samples by thin layer chromatography and gas chromatography. The most abundant polyunsaturated and saturated fatty acids in normozoospermic samples were docosahexaenoic acid (DHA 22 : 6 omega3, 98.5 +/- 4.5 nmol per 10(8) spermatozoa, mean +/- SE) and palmitic acid (103 +/- 17 nmol per 10(8) spermatozoa) respectively. The mean +/- SE values of DHA and palmitic acid in asthenozoospermic samples were 53.9 +/- 11.6 and 145 +/- 14.7 nmol per 10(8) spermatozoa respectively. Compared with normozoospermic samples, asthenozoospermic samples showed lower levels of PUFA and higher amount of saturated fatty acids. The mean +/- SE ratios of sperm PUFA/saturated fatty acids in asthenozoospermic and normozoospermic samples were 0.66 +/- 0.06 and 1.45 +/- 0.16 (P < 0.001) respectively. This study demonstrates that spermatozoa of asthenozoospermic men have lower levels of PUFA compared with saturated fatty acids. This may be contributory to the poor motility noted in samples from these men.