Melanocortin-4 receptor and leptin as genes for the selection of superior Madrasin cattle

Background and Aim: The genetic improvement of cattle through livestock section is based on quantitative, qualitative, and molecular characteristics. This study examined polymorphisms of the melanocortin-4 receptor (MC4R) and leptin genes as a reference for the selection of superior breeds in Madrasin cattle. Materials and Methods: The leptin and MC4R genes of Madrasin cattle were amplified using polymerase chain reaction (PCR); then, restriction fragment length polymorphism of the leptin gene was performed using the restriction enzyme BsaA1, at site 2793 with ACGT point position. Results: The leptin gene was divided into three bands, namely, AA with one fragment (522 bp), CG with two fragments (441 bp and 81 bp), and AG with three fragments (522 bp, 441 bp, and 81 bp). The MCR-4 gene was divided into three bands, namely, 493 bp, 318 bp, and 175 bp. Conclusion: The MC4R and leptin genes can act as molecular markers for growth traits in Madrasin cattle and can be used to genetically optimize and improve growth. The GG allele of the MC4R gene and the AA allele of the leptin gene can be used in Madrasin cattle.

Microbial Diversity and P Content Changes after the Application of Sewage Sludge and Glyphosate to Soil

Pesticides, despite their side effects, are still being used in almost every agriculture, horticulture, maintaining municipal greenery in urban areas and even in home gardens. They influence human life and health and the functioning of entire ecosystems, including inanimate elements such as water and soil. The aim of the study was the evaluation of the suitability of sewage sludge in improving the quality of soil treated with a non-selective herbicide-glyphosate, applied as Roundup 360 SL. A pot experiment was conducted with the use of two arable soils (MS and OS), which were amended with sewage sludge (SS), glyphosate (GL) and sewage sludge with glyphosate (SS+GL). Soil samples were taken after 24 h, 144 h and 240 h and total phosphorus (TP) content (TP), total number of bacteria/fungi, activity of dehydrogenases (Dha), acidic phosphatase (Acp), alkaline phosphatase (Alp), genetic biodiversity of bacteria/fungi using the terminal restriction fragment length polymorphism method were determined. The application of SS and GL to OS caused an increase in Acp (approximately 35%) and a decrease in Alp activity (approximately 20%). Additionally, GL may influence on an increase in the number of fungi and the decrease in the number of bacteria. In soil with SS+GL increase in the fungal diversity in MS and OS was also observed. Moreover, a positive between TP and the number of bacteria and the activity of phosphatases correlation was reported. The obtained results indicate that analyzed sewage sludge could be potentially applied into soil in in situ scale and could constitute a valuable reclamation material.

Genetic polymorphism of ALDH2 in Indonesia’s Minang ethnic

Background: In some people, acetaldehyde, a toxic product from ethanol oxidation, cannot be oxidized to acetate. The excess of acetaldehyde could cause facial flushing, dizziness, and hypertension when they consume ethanol. This ethanol sensitivity is caused by a deficiency of ALDH2. Objective: This study aims to analyze and count the polymorphism frequency of the ALDH2 gene in Indonesia’s Minang ethnic. Methods: DNA samples were taken randomly from hair bulbous of 60 subjects (male and female, 3rd generation). A nested polymerase chain reaction was conducted to amplify the ALDH2 in the samples. Afterward, restriction fragment length polymorphism (RFLP) was conducted to the amplicons using the EcoRI restriction enzyme. The measured parameters were the distribution of the wildtype, atypical homozygote, and heterozygote. Results: Results showed that out of 60 subjects, 53.33% have an atypical homozygote gene (subjects prone to hypersensitive to alcohol), 28.33% have a heterozygote gene, and 18.33% have a wildtype gene. The frequency of the atypical alleles in Minang ethnic is 0.675. Conclusion: The atypical ALDH2 allele was much higher than the normal ALDH2 allele, in which most participants have atypical homozygote ALDH2, suggesting the samples are sensitive to alcohol.

Salivary Microbiota Is Significantly Less Diverse in Patients with Chronic Spontaneous Urticaria Compared to Healthy Controls: Preliminary Results

Background: Because of the important role in regulating the immune system, increasing evidence suggests a possible implication of gut microbiota in Chronic spontaneous urticaria (CSU). Although the oral cavity is the first site of contact between microbiota and the immune system, the association between salivary microbiota and CSU has not yet been reported. Objective: This case-control study aimed to compare differences in salivary microbiota between CSU patients and healthy controls (HC). Twenty-three participants—13 patients with CSU and 10 HC were enrolled; salivary microbiota was determined by molecular approach targeting 16S ribosomal RNA. Terminal restriction fragment length polymorphism (T-RFLP) analysis was performed. Results: Alpha diversity of salivary microbiota in CSU patients was significantly reduced compared to HC, resulting in alteration of the community composition. Species richness determined via the Shannon index was significantly reduced in the CSU group. Conclusion: Dysbiosis of salivary microbiota may contribute to a dysregulated immune system in the development of CSU. To our knowledge, this was the first study that reported an alteration in salivary microbiota composition in CSU patients.

The association of cytochrome 7A1 and ATP-binding cassette G8 genotypes with type 2 diabetes among Jordanian patients

Objectives Increased cholesterol levels were found to be associated with diabetes mellitus type II (DM2). The cholesterol is metabolized by cytochrome 7A1 (CYP7A1) and transported in the intestine by ATP-binding cassette G8 (ABCG8). Genetic variants in CYP7A1 and ABCG8 genes can affect the cholesterol levels. The aim of this study is to compare the frequency of CYP7A1 rs3808607 and ABCG8 rs11887534 and rs4148217 genotypes between healthy and DM2 subjects from Jordanian population. Methods A total of 117 DM2 patients and 100 healthy controls, of Jordanian Arabic origin, were genotyped for CYP7A1 rs3808607 and ABCG8 rs11887534 and rs4148217 genetic variants using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism technique. Results The study showed that homozygosity of rs3808607 (A-204C) genotype in CYP7A1 was significantly higher in DM2 patients (ANOVA, p<0.05) with an odd ratio of 2.66, but rs11887534 (G55C) and rs4148217 (C1199A) genetic polymorphisms in ABCG8 were found in comparable frequencies in both healthy and DM2 subjects. Conclusions The results of this study indicate that CYP7A1 rs3808607 genetic polymorphism is associated with DM2. Further clinical studies are required to confirm this finding among DM2 patients of Jordanian origin.

RELATIONSHIP BETWEEN RFC GENE VARIANTS (RS1051266) AND CLINICAL FEATURES OF SEVERE INTRAVENTRICULAR HEMORRHAGE IN PRETERM INFANTS

The physiological function of the RFC gene is to ensure the processes of intracellular folic acid transport that are extremely important not only for the processes of reproduction, but also for the embryonic develop-ment of a growing foetus. The aim of this study is to investigate the association between the RFC (G80A, rs1051266) gene variants and the features of the neonatal course in premature infants with severe intraven-tricular haemorrhage. Materials and methods. The study included 24 preterm infants with severe intraven-tricular haemorrhages that received standard clinical, laboratory and instrumental examination. Determina-tion of RFC gene variants was performed using the polymerase chain reaction- restriction fragment length polymorphism method. Results. The following frequencies of genotypes were determined according to the G80A variant of the RFC gene: GG – 8 (33.3%), GA – 9 (37.5%) and AA – 7 (29.2%) in the study group. Children with the A allele of the RFC gene had an increased risk of developing hypertension and a higher mean concentration of ionized calcium. Preterm infants with the A allele were more likely to require oxygen therapy with maximum oxygen concentration and in the presence of this allele in a homozygous state (AA genotype) required nCPAP and non-invasive mechanical ventilation. Conclusion. The obtained results sup-port a hypothesis about the influence on variants of RFC genes on severe neonatal period in preterm infants with intraventricular haemorrhages. However, further multifaceted research in this area is required to give more ground in supporting hypothesis.

CD24 GENE POLYMORPHISMS IN EGYPTIAN PATIENTS WITH COLORECTAL CANCER

Introduction: Early detection of colorectal cancer (CRC) is the key to reduce its related morbidity and mortality. Methods: One hundred CRC patients were included in this study, they were admitted at Mansoura Gastroenterology Center, Mansoura University and diagnosis was confirmed by pathology.100 apparently healthy individuals were served as the control group.CD24 (P170 C/T) and CD24 (P-534 A/C) single nucleotide polymorphisms (SNPs) were analyzed using polymerase chain reaction–restriction fragment length polymorphism (PCR/RFLP). Results: Regarding CD24 P170 C/T SNP, no statistically significant association was found between genotypes, alleles and risk of CRC. While, P-534 A/C; AC, CC, AC+CC genotypes and C allele showed significantly higher frequency in cases when compared to controls. CC and TC haplotypes were significantly associated with risk of CRC when compared to control group. Multivariate logistic regression analysis revealed that higher CEA and P-534 A/C (AC+CC) were considered as independent predictors of higher CRC stages. Conclusion: The current study observed an association between CD24 P-543 A/C SNP and the risk of developing CRC in Egyptian subjects.

GENETIC VARIABILITY IN Sechium spp. (Cucurbitaceae) EVALUATED WITH AFLP MARKERS

There are few studies in Mexico aimed at evaluating the genetic variability of Sechium spp. Despite certain biological variants are reported with very high potential to develop antineoplastic supplements to treat public health conditions. Using the Amplified Fragment Length Polymorphism (AFLP) technique, the genetic variability of a sample of 95 accessions of three species of Sechium (S. edule, S. chinantlense, S. compositum) was evaluated, with leaf DNA from the Banco Nacional de Germoplasma de Sechium edule en Mexico. Four combinations of AFPL were applied (EcoRI + ACC/MseI + CAC, EcoRI + ACC/MseI + CAT, EcoRI + ACC/MseI + CGC, and EcoRI + ACC/MseI + CGG). DNA samples were classified into three groups based on the flavour of the fruit (sweet, neutral, bitter). An average of 47.91% polymorphism, 0.16 heterozygosity, 32.83 number of polymorphic bands, and a zero Wright fixation index (Fst) was obtained. The evidence showed that the domesticated accessions (sweet, neutral) were separated from the bitter-taste genotypes. A monophyletic tree was generated with the genetic distance matrix and the neighbour-joining methodology. Analyses showed S. edule as the root taxon, deriving S. compositum and S. chinantlense as subgroups, and suggesting that there is not enough differentiation to treat them as separate species. The evaluated sample showed that there is no apparent reproductive barrier for genetic cross breeding. Genotypes behaved as a complex with evolutive dynamism; that genetic complexity would allow the design of new variants.

REDigest: a Python GUI for In-Silico Restriction Digestion Analysis of Genes or Complete Genome Sequences

Restriction fragment length polymorphism (RFLP) is a technology for the molecular characterization of DNA and widely used genome mapping, medical genetics, molecular microbiology and forensics etc. Terminal restriction fragment length polymorphism (T-RFLP), a variant of RFLP is extensively used in environmental microbiology for the microbial community profiling based on the restriction digestion profile of marker gene (16S rRNA, FTHFS etc.) amplicons. At present, there is a lack of a tool which can perform in-silico restriction digestion of a large number of sequences at a time, in an interactive way and as an output produce sequences of the restriction fragments and visualization plot. I have developed a graphical user interface based software REDigest for the in-silico restriction digestion analysis for gene or genome sequences. The REDigest software program with a graphical user interface is freely available at https://github.com/abhijeetsingh1704/REDigest.

The Association of the Phylogenetic Typing of the Klebsiella pneumoniae Isolates with Antibiotic Resistance

Klebsiella pneumoniae complex (KPC) accounts for approximately one-third of all Gram-negative infections. Moreover, it is highly resistant and can taxonomically be distributed into KpI, KpII, and KpIII phylogroups. This study aimed to investigate the distribution of phylogenetic groups and the relationship between them and antibiotic resistance patterns. For this purpose, we collected KPC isolates from Tabriz, Iran, between 2018 and 2020. Antimicrobial susceptibility testing was performed by disk diffusion agar, and phylogenetic groups were then examined using gyrA restriction fragment length polymorphism (RFLP) and parC PCR methods. A total of 100 KPC isolates were obtained from the clinical specimens (urine, respiratory secretion, blood, wounds, and trachea). The enrolled patients included 47 men and 53 women aged from 1 to 91 years old. The highest sensitivity was found related to fosfomycin as 85%, followed by amikacin as 66%. The three phylogenetically groups by the RFLP-PCR method were found in KPC, 96% (96 isolates) as KpI, 3% (3 isolates) as KpII, and 1% (1isolate) as KpIII. The highest antibiotic resistance was observed in KpI. It was shown that a valid identification of three phylogenetic groups of KPC can be done by combining both gyrA PCR-RFLP and parC PCR. Of note, the KpI group was also observed as the dominant phylogenetic group with the highest resistance to antibiotics.