Primary antibody deficiencies in Turkey: molecular and clinical aspects

Primary antibody deficiencies (PAD) are the most common subtype of primary immunodeficiencies, characterized by increased susceptibility to infections and autoimmunity, allergy, or malignancy predisposition. PAD syndromes comprise of immune system genes highlighted the key role of B cell activation, proliferation, migration, somatic hypermutation, or isotype switching have a wide spectrum from agammaglobulinemia to selective Ig deficiency. In this study, we describe the molecular and the clinical aspects of fifty-two PAD patients. The most common symptoms of our cohort were upper and lower respiratory infections, bronchiectasis, diarrhea, and recurrent fever. Almost all patients (98%) had at least one of the symptoms like autoimmunity, lymphoproliferation, allergy, or gastrointestinal disease. A custom-made next-generation sequencing (NGS) panel, which contains 24 genes, was designed to identify well-known disease-causing variants in our cohort. We identified eight variants (15.4%) among 52 PAD patients. The variants mapped to BTK (n?=?4), CD40L (n?=?1), ICOS (n?=?1), IGHM (n?=?1), and TCF3 (n?=?1) genes. Three novel variants were described in the BTK (p.G414W), ICOS (p.G60*), and IGHM (p.S19*) genes. We performed Sanger sequencing to validate pathogenic variants and check for allelic segregation in the family. Targeted NGS panel sequencing can be beneficial as a suitable diagnostic modality for diagnosing well-known monogenic PAD diseases (only 2–10% of PADs); however, screening only the coding regions of the genome may not be adequately powered to solve the pathogenesis of PAD in all cases. Deciphering the regulatory regions of the genome and better understanding the epigenetic modifications will elucidate the molecular basis of complex PADs.

     

Correction to: Primary antibody deficiencies in Turkey: molecular and clinical aspects

Immunologic Research -     

Assessment of cardiac parameters in evaluation of cardiac functions in patients with thalassemia major

The aim of the study was to evaluate cardiac function and early cardiac dysfunction of patients followed as thalassemia major. In this study, the authors compared 100 patients, diagnosed as thalassemia major with mean age 11.84 ± 4.35, with 60 healthy control subjects at the same age between 2008 and 2011. Early diagnosis of iron overload that may occur after repeated transfusions is important in this patient group. To detect early iron accumulation, the authors compared ferritin with the echo findings, the 24-hour Holter, and cardiac magnetic resonance imaging (MRI) T2* values in the patients of same age and sex, treated with chelators, without heart failure, nonsplenectomized, and do not differ in the presence of hepatitis C. Ferritin levels, left ventricular systolic functions (ejection fraction [EF], shortening fraction [SF]), left ventricular measurements, left ventricular diastolic functions, T2* image on cardiac magnetic resonance, heart rate variables in 24 hours, and Holter rhythm were evaluated to show the early failure of cardiac functions. In this study the authors confirmed that iron-related cardiac toxicity damages electrical activity earlier than myocardial contractility. Left ventricular diastolic diameter (LVDd), left ventricular mass (LVM), and LV systolic diameter (LVDs) levels were significantly higher in the patient group with ectopia. Patients with ectopia are the ones in whom LVM and LVDd are increased. In thalassemia major patients with ectopia, LF/HF ratio was markedly increased, QTc dispersion was clearly found higher in patients with ectopia rather than nonectopic patients. The standard deviation all normal RR interval series (SDNN) was found clearly lower in thalassemia major group with ectopia than control group because it is assumed that increase in cardiac sympathetic neuronal activity is related to exposure to chronic diastolic and systolic failure.     

Correlation of Plasma B-Type Natriuretic Peptide with Shunt Severity in Patients with Atrial or Ventricular Septal Defect

The goal of this study was to test the utility of bedside plasma concentration of B-type natriuretic peptide (BNP) assay as a screen for large shunts in pediatric patients with atrial septal defects (ASDs) and ventricular septal defects (VSDs). Thirty-five children at a mean age of 70 ± 129 weeks with ASD or VSD were included in the study. Nine patients had VSD and 26 had ASD. Plasma BNP values were compared with the Q _p/Q _s ratios derived from quantitative Doppler flow measurements. Mean BNP was 29 ± 42 pg/ml, with a range between <5 pg/ml and 208 pg/ml. Sixteen patients had Q _p/Q _s values >1.5 and 19 had values <1.5. The difference of mean BNP in these patient groups was statistically significant (45 ± 56 vs 14 ± 17, p = 0.03). BNP was positively correlated with shunt significance. Receiver operating characteristic curve analysis revealed a sensitivity of 69% and a specificity of 79% at a plasma BNP cut-off level of ≥20 pg/ml. Bedside measurement of BNP correlates with magnitude of ASD and VSD in children. BNP can provide information for the management of children with ASD or VSD. It can be used as part of the evaluation of a child with a preliminary diagnosis of a congenital defect.     

Effect of lipopolysaccharide on circadian clock genes <Emphasis Type="Italic">Per2</Emphasis> and <Emphasis Type="Italic">Bmal1</Emphasis> in mouse ovary

In mammals, circadian rhythms are associated with multiple physiological events. The aim of the present study was to examine the effect of lipopolysaccharide (LPS) on circadian systems in the ovary. Immature female mice were received an intra-peritoneal injection of equine chorionic gonadotropin (eCG) and LPS. Total RNA was collected from the ovary at 6-h intervals throughout a 48 h of experimental period. The expression of the circadian genes period 2 (Per2) and brain and muscle ARNT-like 1 (Bmal1) such as circadian genes was measured by quantitative PCR. Although expression of Per2 and Bmal1 in the ovary did not display clear diurnal oscillation, LPS suppressed the amplitude of Per2 expression. Additionally, LPS inhibited the expression of cytochrome P450 aromatase (CYP19) and luteinizing hormone receptor (LHr) genes in the ovary of eCG-treated mice. Our data suggest that Per2 may be associated with the inhibition of CYP19 and LHr expression by LPS in the ovaries of immature mice.     

Immunohistochemical expression of PTEN in normal, hyperplastic and malignant endometrium and its correlation with hormone receptors, bcl-2, bax, and apoptotic index

PTEN is a tumor suppressor gene that is frequently mutated in type I endometrioid endometrial carcinomas (EECs), and is involved in the control of cell proliferation, differentiation, and apoptosis. In this study, we aimed to assess the relationship between PTEN expression and estrogen, progesterone receptors (PRs), other apoptosis-related proteins, such as bcl-2 and bax, and apoptotic index (AI) in EEC, its precursor lesion hyperplasia, and cyclical endometrium. We also evaluated the relationship between PTEN expression and clinicopathologic parameters. PTEN, estrogen receptor (ER), PR, and bcl-2 and bax expressions were evaluated immunohistochemically, and AI was evaluated in hematoxylin and eosin (HE)-stained slides in 23 cyclical and 37 hyperplastic endometria and in 35 EECs. PTEN expression was higher in cyclical endometrium than in the carcinomas (p<0.05). The PTEN expression level was significantly higher in non-atypical hyperplasias than in EEC, but there were no differences between atypical complex hyperplasia (ACH) and EEC and between hyperplasias. In the carcinomas, there was a negative correlation between grade and PTEN expression (r=-0.338, p=0.047). In conclusion, we presume that PTEN is involved in the early phases of endometrial tumorigenesis, and it can be speculated that decreased PTEN expression with loss of differentiation in carcinoma can contribute to the emergence of tumors with a more aggressive phenotype.     

Lipid peroxidation, erythrocyte superoxide-dismutase activity and trace metals in young male footballers

Physical training is known to induce oxidative stress in individuals subjected to intense exercise. In this study, we investigated plasma malondialdehyde (MDA) levels and erythrocyte superoxide dismutase (SOD) activity of 25 young male footballers and a control group of similar age. Red blood cell (RBC) count, haemoglobin (Hb) and haematocrit (Hct) values, and copper (Cu) and zinc (Zn) levels were also examined. The maximal oxygen uptake (VO2max) of all subjects was determined in order to establish their functional capacity. The main finding of the present study was that plasma MDA levels, one of the most commonly used markers of lipid peroxidation, of this group of footballers aged under 21 decreased slightly when compared with those of the control group (p < 0.001). In contrast, erythrocyte SOD activity was higher in the footballer group than in the controls (p < 0.001). Footballers who are under regular training showed an improved antioxidant activity in comparison to sedentary controls. Plasma copper concentration, RBC count and Hb concentration of the footballer group were all significantly lower than those of the control group, (p < 0.001, p < 0.01, p < 0.01, respectively). Investigating the footballers' data with Spearman's correlation analyses, the correlation coefficients (r) between Zn/Cu ratio and SOD was positive (r=0.44; p < 0.05); and between VO2max and SOD (r=0.42; p < 0.05) were both positive. On the basis of statistical analysis, we suggest that regular exercise may be beneficial in cases of oxidative damage by reducing the amount of lipid peroxidation and increasing the activity of the antioxidant enzyme SOD.     

Acute effect of intranasal estrogen on cerebral and cerebellar perfusion in postmenopausal women

OBJECTIVES: Estrogen action in the brain influences many neurochemical processes. The aim of the study was to evaluate the acute effect of intranasal 17beta-estradiol on cerebral and cerebellar perfusion in postmenopausal women. METHODS: The study group included 24 healthy postmenopausal women who had been in natural menopause for at least 1 year (mean age: 47.38+/-5.9 years). We conducted an experimental, randomized, placebo-controlled, cross-over, double-blind study. Cerebral and cerebellar perfusion was measured after placebo (saline serum physiologic) or intranasal 17beta-estradiol administration by Single Photon Emission Computed Tomography (SPECT) using technetium-99m-hexamethylpropylene amine oxime (Tc99m-HMPAO). Regions of interest (ROIs) were drawn manually. Cerebral and cerebellar perfusions were calculated for each ROI using average number of counts per pixel. Semiquantitative analysis was performed in bilateral frontal, temporal, parietal and occipital lobes, thalamus, putamen, hippocampus, amygdala, caudate nuclei, cerebellar region, anterior/posterior of cingulate gyrus and pons. RESULTS: After intranasal 17beta-estradiol administration, SPECT study revealed significant increases in cerebral and cerebellar perfusion compared to placebo measurements in all studied slices (p<0.05). There was a positive correlation between serum estrogen levels after 17beta-estradiol and cerebral and cerebellar perfusion. CONCLUSIONS: Administration of single dose intranasal 17beta-estradiol increases cerebral and cerebellar perfusions in healthy postmenopausal women.     

Analysis of UV-stimulated recombination in the Drosophila SMART assay

The UV component of solar radiation is classified into UVA (320-400 nm), UVB (290-320 nm), and UVC (200-290 nm). Although all three types of UV light are capable of damaging biological systems, the earth's atmosphere filters out UVC, and a portion of UVB. In this study, we evaluated the induction of mutation and recombination by different wavelengths of UV light in the wing spot test of Drosophila melanogaster (Somatic Mutation and Recombination Test, SMART). Third-instar larvae that were trans-heterozygous for the third chromosome recessive markers, multiple wing hairs (mwh) and flare-3 (flr(3)), were exposed to different doses of UVA (at 365 nm), UVB (at 312 nm) or UVC (at 254 nm), and transferred to standard Drosophila culture medium. Feeding ended with pupation of the surviving larvae, and the genetic changes induced in the somatic cells of the wing's imaginal discs lead to the formation of mutant clones on the wing blade. Point mutation, chromosome breakage, and mitotic recombination produce single spots, while twin spots are produced only by mitotic recombination. Exposure to 500-4,000 J/cm(2) UVA did not increase the frequency of mutant spots. UVB doses of 200, 250, 300, 350, and 400 J/cm(2) increased the frequency of all categories of spots, indicating that UVB was potentially both mutagenic and recombinogenic. Assays run in balancer-heterozygous flies (which are insensitive to recombination) indicated that the fraction of mutants in trans-heterozygous flies due to recombination increased from 48.57% at 200 J/cm(2) UVB to 98.30% at 400 J/cm(2) UVB. While 140-480 J/cm(2) of UVC was not genotoxic, UVC produced a strong toxic response at doses higher than 140 J/cm(2). The results of this study indicate that UVB was much more active than UVC or UVA in the SMART assay, and that UVB was highly recombinogenic.