Molecular Characterization of Cytidine Monophospho-N-Acetylneuraminic Acid Hydroxylase (CMAH) Gene and Frequency of Blood Types in Stray Cats of İzmir, Turkey

Cytidine monophospho-n-acetylneuraminic acid hydroxylase (CMAH) gene associated with blood groups in cats encodes CMAH enzyme that converts Neu5Ac to Neu5Gc. Although variations in CMAH gene of pedigree cats have been revealed, the presence/lack of them in non-pedigree stray cats is unknown. Therefore, the present study aimed to investigate the variations in CMAH gene and the quantity of Neu5Ac and Neu5Gc on erythrocytes of non-pedigree stray cats (n:12) living in İzmir, Turkey. Meanwhile, these 12 cats were typed using the mitochondrial DNA control region. Also, the frequency of blood types was determined in 76 stray cats including 12 cats that were used for CMAH and Neu5A/Neu5Gc analysis. In total, 14 SNPs were detected in 5’UTR as well as in exon 2, 4, 9, 10, 11 and 12 of CMAH gene. Among these SNPs, -495C>T in 5’UTR was detected for the first time as heterozygous in type A and AB cats, and homozygous and heterozygous in type B cats. The remaining 13 that have been detected in previous studies were also found as homozygous or heterozygous. Homozygous form (T/T) of the -495C>T polymorphism was found among only type B cats. Among the polymorphisms previously determined in the literature, homozygous form of the -371C>T polymorphism was found among only type B cats whereas heterozygous form (A/C) of the 327A>C polymorphism was detected in only type AB cats. Both Neu5Gc and Neu5Ac were detected in type A and AB cats. In type B cats, only Neu5Ac was detected. Among two type AB cats, the level of Neu5Ac was found higher in cat carrying heterozygous form (T/C) of 1392T>C. Mitotypes A, A6, D, E and 1 were detected among stray cats analysed for the characterisation of CMAH gene. The prevalence of type B cats (67.1%) was higher than others. As a result, the presence of a new SNP as well as previous SNPs indicates that more variations can be found in stray cats with a more comprehensive study in the future. Also, the high prevalence of type B cats demonstrates the high risk of neonatal isoerythrolysis among stray cats living in İzmir, Turkey.

Identification of p.His119Leu mutation in the G6PC gene of a Vietnamese patient with glycogen storage disease type Ia

Glycogen storage disease type Ia (GSD Ia), a rare autosomal inherited disorder, is characterized by accumulation of excessive glycogen and fat in the liver. Primary symptoms of GSD Ia include hypoglycemia; metabolic acidosis; elevated levels of lactate, uric acid and lipids; hepatomagaly and growth retardation. Glycogen storage disease type Ia was caused by mutations in the G6PC gene. In this study, mutations in a Vietnamese patient with glycogen storage disease type Ia were analyzed using the whole exome sequencing method. A missense mutation c.356A>T (p.His119Leu) in the G6PC gene of the patient was identified in exon 3. Genetic analysis confirmed that this mutation was present under homozygous form In-silico analyses using SIFT and Mutation Taster confirmed the damaging effects of this mutations on the function of the proteins. This result enriches knowledge of the G6PC gene mutation spectrum and provides genetic data for further studies on glycogen storage disease type Ia in Viet Nam.

ВИЗНАЧЕННЯ ГЕНЕТИЧНОГО ПОЛІМОРФІЗМУ CYP2C9 НА ТЛІ ПРИЗНАЧЕННЯ ВАРФАРИНУ У ПАЦІЄНТІВ НА ІШЕМІЧНУ ХВОРОБУ СЕРЦЯ, УСКЛАДНЕНУ ПОСТІЙНОЮ ФОРМОЮ ФІБРИЛЯЦІЇ ПЕРЕДСЕРДЬ

This article describes the results of a genetic study of the spread of CYP2C9 polymorphic variants in patients in the Podilskyi region of Ukraine. In different ethnic groups, the frequency of cytochrome polymorphic isoenzymes can vary significantly. The presence of genetic mutations of this cytochrome (Arg144Cys, Ile359Leu) is associated with an increased concentration of warfarin in the blood, which can lead to excessive hypocoagulation and risk of bleeding. Particular attention should be paid to patients with a detected mutation in homozygous form, as both alleles of the gene are mutant (their carriers are "slow metabolizers" and therefore require more careful dose selection). Patients are advised to undergo individual genotyping, which will help to predict the risk of each individual patient.

Assessment of the occurrence of gene polymorphisms CAPN316 and UoGCAST in the population of cattle

To solve the problem of increasing the rate of breeding, but reduce the breeding work with farm animals, it is necessary to form herds with the desired level of productivity, that are adapted to specific regions of breeding, resistant to various diseases, with a decrease in the time for the breeding process. The active use of molecular genetic methods has contributed to the expansion of the list of DNA markers for farm animals, which are candidate genes for economically useful traits. Among these genes are widely known members of the calpain-calpastatin system, which is associated with postmortem proteolysis and tenderisation of muscles. The calpain system consists of an actively expressed μ-calpain (CAPN1) and m-calpain (CAPN2) and a single endogenous inhibitor, CAPN1 and CAPN2-calpastatin (CAST). The study of polymorphisms of these genes contributes to the expansion of marker characterisation in breeding. DNA samples (n=139) obtained from the blood of cattle were used in the work. Real-time PCRs were carried out using a ANK-32 programmable thermocycler (Synthol, Moscow, Russia). The CAPN316 gene polymorphism was present in 15% of animals tested, with allele G being found in 85% of animals. Similar calculations on the occurrence of the UoGCAST gene polymorphism in this sample of animals found the desirable allele G in 22% of animals and allele C in the remaining 78%. From the analysis of the occurrence of both genes and their polymorphisms in the study population, animals that had a combination of both desirable genotypes (CC*GG*) of CAPN316 and UoGCAST genes were not identified. There were also no animals that had the desired CAPN316 CC genotype and the heterozygous state of the UoGCAST gene (CC*GC). While 7.6% of the animals were CC*GG for the desirable expression of gene CAPN316, they contained the homozygous form of the gene UoGCAST.

Investigation of some endogenous antimicrobial peptides in thalassemia

The aim of this work was a comparative study of the amount of antimicrobial peptides - human neutrophil peptides - defensins (HNP), hepcidin, bactericidal/permeability-increasing protein (BPI), and endotoxin in β-thalassemia. Blood samples of 135 patients with thalassemia were investigated. All patients were divided into 3 groups. The first group included patients with heterozygous form (n=45). The second group consisted of patients with homozygous form before splenectomy (n=45). The third group included patients with homozygous form after splenectomy (n=45). The age of patients varied from 2 to 18 years. Biochemical [unconjugated and conjugated bilirubin, alkaline phosphatase, hemoglobin, ferritin, aspartate transaminase (AST), alanine transaminase (ALT), mean corpuscular volume (MCV)] and immune (IgA, IgM, IgG, phagocytic activity) parameters were defined. Obtained results suggest that increased levels of endogenous antimicrobial peptides are associated with the development of the infectious process and reflect the dynamics of changes in biochemical parameters and immune status.

RISK FACTORS OF BONE MINERAL DENSITY DEFICIT AND LOW-ENERGY FRACTURES IN PRIMARY OSTEOPOROSIS IN MEN

Purpose:to evaluate the role of well-known factors on the formation of bone mineral density (BMD) and risk of fractures in primary osteoporosis in men.Patients and methods.The influence of well-known osteoporosis risk factors such as age, smoking, alcohol consumption, obesity, genetic disorders in genes encoding COL1A1, COL1A2 and VDR-receptor products, history of low-energy fractures in first-line relatives upon the BMD value and risk of fractures was evaluated in 231 patients with primary osteoporosis. All patients were divided into three age groups according to the following forms of osteoporosis: 17-20 years (n=26) – juvenile form, 21-50 (n=103) – idiopathic form, patients over 51 years (n=102). To assess the influence of study factors on the risk of fractures the patients were subdivided into 5 groups according to fracture localization.Results.The relationship between BMD deficit and mutations in homozygous form of gene rs2412298 (encodes collagen), and the tendency to a reliable increase of BMD deficit in L1 – L4 vertebrae under polymorphism in homozygous form of 1800012 gene was noted that might be evidence of their contribution to the development of primary osteoporosis in men. At the same time, smoking, alcohol consumption, age, mutations in homozygous form of gene rs2412298 and a history of low-energy fractures in first-line relatives increased the risk of low-energy fractures of the vertebral bodies and proximal femur.

Risk factors of bone mineral density deficit and low-energy fractures in primary osteoporosis in men

Purpose: to evaluate the role of well-known factors on the formation of bone mineral density (BMD) and risk of fractures in primary osteoporosis in men. Patients and methods. The influence of well-known osteoporosis risk factors such as age, smoking, alcohol consumption, obesity, genetic disorders in genes encoding COL1A1, COL1A2 and VDR-receptor products, history of low-energy fractures in first-line relatives upon the BMD value and risk of fractures was evaluated in 231 patients with primary osteoporosis. All patients were divided into three age groups according to the following forms of osteoporosis: 17-20 years (n=26) - juvenile form, 21-50 (n=103) - idiopathic form, patients over 51 years (n=102). To assess the influence of study factors on the risk of fractures the patients were subdivided into 5 groups according to fracture localization. Results. The relationship between BMD deficit and mutations in homozygous form of gene rs2412298 (encodes collagen), and the tendency to a reliable increase of BMD deficit in L1 - L4 vertebrae under polymorphism in homozygous form of 1800012 gene was noted that might be evidence of their contribution to the development of primary osteoporosis in men. At the same time, smoking, alcohol consumption, age, mutations in homozygous form of gene rs2412298 and a history of low-energy fractures in first-line relatives increased the risk of low-energy fractures of the vertebral bodies and proximal femur.