T1 Genotypes Research Articles

Functional characterization of chalcone reductase gene CHR3 through RNAi

In this experiment, the soybean genome "Jilin 30" was used as a template to clone the target gene CHR3 using specific primers. The sense fragment, antisense fragment, and intron of the target gene were ligated into pCAMBIA3300 through double enzyme digestion to form a stem loop structure. We then transferred the constructed RNAi vector pCAMBIA3301-CHR3 RNAi into the recipient soybean genome via Agrobacterium mediated method, specifically reducing the expression of the target gene CHR3. By genetic transformation, positive plants were obtained, and molecular biology methods such as Southern blotting, real-time quantitative PCR, isoliquiritigenin analysis, and identification of resistance to Phytophthora root rot were used to detect the expression level of the CHR3 gene in the positive plants and study its function.The RNAi expression vector pCAMBIA3300-CHR3-RNAi was successfully constructed using the sense and antisense fragments of 428 bp CHR3 gene and an intron of 110 bp was inserted to form the stem-loop structure of the RNAi vector. PCR was used to detect 6 lines of the T1 generation and 15 lines of the T2 generation. The results of Southern analysis confirmed the integration of CHR3 and marker genes into the soybean genome. The expression CHR3 gene in the T1 genotypes decreased by 13.1 to 58.3%; the expression in the root decreased by 0.9 to 47.4%; the expression in the leaf decreased by 20 to 44.2%; and the expression in the stem decreased by 7.6 to 23%. The content of isoliquiritigenin decreased by 26.7% as transgenic shoots had a lower content of isoliquiritigenin and reduced resistance to Phytophthora sojae. Bangladesh J. Bot. 53(3): 627-633, 2024 (September) Special

Wheat Yield and Important Traits Influenced by Interaction of Potassium and Irrigation Levels Evaluated at Number of Locations in the Country by AMMI Analysis and Non-Parametric Measures

AMMI analysis of treatments consisted of levels of potassium with irrigations observed highly significant effects of locations, treatments, and T×L interactions for wheat yield. About 53.4% of the total variations in yield values was due to locations followed by 26.3% and 10.8% by treatments and interactions effects. Further analysis found 59.7% contributed by AMMI1 while 17.2% and 9.4% by AMMI2 and AMMI3 components for thousands grain weight as total of first two components cumulative to 76.9% of the total variation. The sums of squares for G×E signal and noise were 56.7% and 43.3% of interaction effects for grains per spike as the sum of squares of T×L signal was 2.58 times of treatments effects and IPC1 alone was 3.54 times the treatments effects. Measures ASV and ASV1 recommended T6, T5, T4 for wheat yield while measures utilized 81.6% of interaction sum of squares whereas MASV and MASV1 measures based on 98.4% identified T3, T5, T8, T4 treatments. Maximum average for thousands grains weight; GAI selected T8, T9, T6 whereas as per HM values treatments T5, T2, T8 would be more desirable. Grains per spike found the measures RPGV and HMRPGV settled for T8, T9, T5 treatments. Non parametric measures for yield observed Si1 selected T3, T2, T5 as opposed to T6, T4, T1 by Si2 values. T6, T4, T1 genotypes considered by Si3 Si4 measure considered T6, T4, T1 next Si5 for T6, T3, T4 and Si6 pointed towards T6, T4, T8 genotypes while Si7 favored T6, T1, T4 genotypes. Composite measures for thousands grains weight found NPi(1) for T3,T4,T7 while as per NPi(2) for T4,T3,T7, NPi(3) T4,T3,T2, NPi (4) found T4, T5, T7 as suitable treatment combinations. Multivariate hierarchical clustering as per Ward’s method for wheat yield observed first irrigation level with three potassium levels formed a cluster and other irrigation levels with potassium application remained in other one. At the first node of demarcation for thousands grains weight IPC5 exhibited MASV with MASV1, ASV1, IPC4, ASV, Si1 Si2 Si3 Si4 Si5 Si6 Si7 NPi(1), CV in one side and mean, GAI, PRVG, IPC1, HM, IPC1, NPi(2) NPi(3) NPi (4) on other side. The performance of treatments based on AMMI and non-parametric measures would be more meaningful for identification of suitable irrigation and potassium levels for wheat sustainable production.

Clinical isolates of Mycobacterium tuberculosis with different genotypes exhibit distinct host macrophage responses in vitro.

Introduction. Mycobacterium tuberculosis (M.tb), the causative agent of tuberculosis, can survive as an intracellular parasite after entering macrophages via phagocytosis. M.tb strains are genotypically distinct and engage in diverse pathogen-host interactions, with different host immune responses triggered by different M.tb strains. Importantly, differences in intracellular accumulation and triggering of host macrophage responses during early infection stages are key determinants that shape the final outcomes of host innate immune responses to different M.tb strains.Hypothesis/Gap Statement. Clinical M.tb strains with different genotypes elicit different host innate immune responses in vitro.Aim. This work aimed to compare host innate immune responses elicited by genotypically diverse, clinically derived M.tb strains in vitro.Methodology. RAW264.7 cells were infected with three lineage 2 and lineage 4 clinically derived M.tb strains and strain H37Rv. Strains were evaluated for differences in intracellular growth, induction of macrophage apoptosis, and induction of expression of proinflammatory cytokines and associated pattern recognition receptors.Results. Highly variable cytokine profiles were observed subsequent to RAW264.7 cell infection with the different strains. The Beijing genotype strain, a modern Beijing strain belonging to lineage 2, induced milder host proinflammatory responses and less apoptosis and exhibited greater intracellular growth as compared to the other strains. Moreover, mRNA expression levels of iNOS in Beijing and MANU2 genotype strains exceeded corresponding levels obtained for the T1 genotype strain. Meanwhile, mRNA expression levels of toll-like receptor (TLR)-encoding genes TLR2 and TLR7 in macrophages infected with the Beijing genotype strain were higher than corresponding levels observed in MANU2 genotype strain-infected macrophages.Conclusion. The higher intracellular survival rate and lower level of host cell apoptosis associated with macrophage infection with the Beijing genotype strain indicated greater virulence of this strain relative to that of the other strains. Furthermore, in vitro immune responses induced by the Beijing genotype strain were unique in that this strain induced a weaker inflammatory response than was induced by T1 or MANU2 genotype strains. Nevertheless, additional evidence is needed to confirm that Beijing genotype strains possess greater virulence than strains with other genotypes.

Glutathione S-transferase genetic polymorphisms and fluoride-induced reproductive toxicity in men with idiopathic infertility.

Male infertility caused by idiopathic oligoasthenospermia (OAT) is known as idiopathic male infertility. Glutathione S-transferase (GST) and fluoride may play important roles in idiopathic male infertility, but their effects are still unknown. Our study examined the relationship between GST polymorphisms and fluoride-induced toxicity in idiopathic male infertility and determined the underlying mechanism. Sperm, blood, and urine samples were collected from 560 males. Fluoride levels were measured by a highly selective electrode method, and GST genotypes were identified using polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). Semen parameters, DNA fragmentation index (DFI), mitochondrial membrane potential (MMP), and oxidative stress (OS) biomarkers were statistically assessed at the P < 0.05 level. Compared with healthy fertile group, semen parameters, fluoride levels, OS biomarkers, sex hormone levels, and MMP and DFI levels were lower in the idiopathic male infertility group. For glutathione S-transferase M1 (GSTM1[-]) and glutathione S-transferase T1 (GSTT1[-]) or glutathione S-transferase P1 (GSTP1) mutant genotypes, levels of semen fluoride, OS, MMP, and DFI were considerably higher, and the mean levels of sperm parameters and testosterone were statistically significant in GSTM1(+), GSTT1(+), and GSTP1 wild-type genotypes. Both semen and blood fluoride levels were associated with oxidative stress in idiopathic male infertility patients. Elevated fluoride in semen with the genotypes listed above was linked to reproductive quality in idiopathic male infertility patients. In conclusion, GST polymorphisms and fluorine may have an indicative relationship between reproductive quality and sex hormone levels, and OS participates in the development of idiopathic male infertility.

Report of rare genotypes of Acanthamoeba from soil source of the Payeh Maga Highland forest, North-eastern Sarawak, Malaysia

Pathogenic Acanthamoeba species are the causative agents of Acanthamoeba keratitis and granulomatous amoebic encephalitis. Members of this amoeba genus are ubiquitous in the environments. In Malaysia, most environmental studies performed to date have targeted the detection and characterisation of Acanthamoeba sp. in different water sources, dust and soil samples collected near human habitats. However, no localstudy has yet to examine these amoebae in a forest, an isolated terrestrial environment, where human activity is relatively scarce. Further, there are also limited studies to investigate the same globally. The current study reported the isolation, morphological and genotypic characterisations of eleven Acanthamoeba sp. isolated from soils of the Payeh Maga Highland forest, Sarawak, Malaysia. Morphological analysis revealed that nine isolates belonged to Group II, whereas two isolates belonged to Group III as defined by the criteria of Pussard and Pons. The phylogenetic analysis based on complete 18S rRNA gene sequences showed that the isolates belonged to the rare T1 (six isolates), T6 (two isolates) and T13 (three isolates) genotypes. To the best of our knowledge, this is the first report about the detection of T6 Acanthamoeba sp. in this country. Overall, the current findings have enriched the knowledge pertaining to the occurrence, morphological and genotypic characteristics of Acanthamoeba sp. in an isolated terrestrial environment of Malaysia.

Effect of glutathione S-transferase gene polymorphisms on semen quality in patients with idiopathic male infertility.

ObjectiveTo investigate the relationship between glutathione S-transferase enzyme (GSTM1, T1, and P1) genetic variants and semen quality in men with idiopathic infertility.MethodsSperm characteristics were measured using computer-assisted sperm analysis. The malondialdehyde (MDA), nitric oxide (NO), and total antioxidant capacity (TAC) activities were detected by spectroscopic analysis, and 8-hydroxy-2′-deoxyguanosine (8-OHdG) was detected by enzyme-linked immunosorbent assay.ResultsThis study included 246 idiopathic infertile men and 117 controls. The GSTM1(−), T1(−), and M1/T1(−/−) genotype frequencies significantly differed between the groups. The GSTM1(−) and T1(−) genotypes in idiopathic infertile men negatively correlated with sperm concentration, motility, mitochondrial membrane potential, and other parameters. However, these genotypes positively correlated with the amplitude of the lateral head displacement and NO and 8-OHdG levels. The GSTT1(−) genotype positively correlated with mean angular displacement and MDA activity. GSTM1(−) and T1(−) had a synergistic effect on semen quality. Sperm motility, normal morphology, straightness, and TAC were lower and amplitude of lateral head displacement and MDA were higher in the GSTP1(A/G + G/G) group than in the GSTP1(A/A) group among men with idiopathic infertility.ConclusionsGSTM1, T1, and P1 genetic variants may be risk factors for infertility by affecting the semen quality men with idiopathic oligoasthenospermia.

Identification of cadmium tolerant and sensitive genotypes of castor and their contrasting responses to cadmium treatment.

Present study identified WM (T1) and DCH-177(T2) as cadmium (Cd) tolerant and GCH2 (S1) and GCH4 (S2) as Cd-sensitive genotypes of castor. Cd treatment (500 μM) led to a significant decline in leaf and root biomass, photosynthetic rate (Pn), stomatal conductance (C), transpiration rate (E), water use efficiency (WUE), photosynthetic pigments content, and altered chlorophyll fluorescence in S1 and S2 genotypes but no significant changes were detected in the T1 and T2 genotypes. The content of H2O2 and malondialdehyde (MDA; stable end product of lipid peroxidation) increased significantly in S1 and S2 castor plants treated with 500 μM Cd, whereas no significant alteration was observed in T1 and T2 in comparison to their respective controls. In general, Cd tolerance is associated with its reduced accumulation. However, no significant difference in Cd accumulation in roots and leaves of S1 genotype was observed compared to T1 genotype. Significant enhancement in the concentration of redox-active metal copper (Cu) was observed in roots and leaves of 500 μM Cd-treated S1 plants but not in T1 plants. Overall, our results suggest that enhanced content of Cu, H2O2, and MDA and reduced photosynthetic parameters might be the cause of Cd sensitivity observed in the S1 genotype.

Frequency of glutathione S-Transferase M1, T1 and P1 genotypes and their combinations in northern West Bengal, India: Implications for disease association

Glutathione S-Transferases (GSTs) are important xenobiotic-detoxifying enzymes in animals. Polymorphisms in GST M1, GST T1, and GST P1 genes are common within human populations and have been linked to several multi-factorial diseases, including carcinogenesis. The present study aimed to determine the genotypic frequencies of M1, T1, and P1 in the healthy individuals (N = 332) of the northern region of West Bengal, India and to compare the frequencies with different populations studied elsewhere. Genotyping of the M1 and T1 alleles was performed using multiplex-PCR and the polymorphism of P1 allele was studied by PCR-RFLP using Alw261 restriction endonuclease. The M1 and T1 null genotypes were found in 34.9% and 27.4% individuals respectively and 9.9% of individuals were found to be double-null genotypes. The genotypic frequency of P1 revealed that 62.2% of individuals had wild type Ile/Ile genotypes, 29% were heterozygous Ile/Val and 8.7% individuals were homozygous Val/Val. There seemed to be no association between the presence of M1 and T1 alleles with any P1 genotypes (p < 0.05). While frequency of the M1 was found to be similar to most of the Indian populations, T1 was found to be different from other Indian populations. M1 and T1 frequencies are found to be similar to North-East Indian population but were higher than that found in other Indian populations. Given the importance of the alleles in detoxification of pesticides, drugs and carcinogenesis and the preponderance of null genotypes in human disease conditions, this study approaches to delineate demographic indicators that may help future investigations in pharmacological, clinical and genetic association studies.

Acanthamoeba species isolated from Philippine freshwater systems: epidemiological and molecular aspects.

Free-living amoeba (FLA) research in the Philippines is still in its infancy but has, by far, demonstrated the presence of potentially pathogenic species. Acanthamoeba may cause sight-threatening and central nervous system infections to humans, yet its epidemiologic distribution from local environmental sources is yet to be defined. The present study aimed to provide a baseline epidemiologic distribution of Acanthamoeba spp. in freshwater systems in the Philippines and establish potential pathogenicity of isolates through thermo-tolerance assay. A total of 63 water samples were collected from 13 freshwater systems all over the Philippine archipelago. The low-volume (50ml) water samples were processed and cultured on non-nutrient agar lawned with Escherichia coli and observed for amoebic growth using light microscopy. Amoebic culture demonstrated 14.28% (9/63) positivity while further molecular testing of culture-positive plates using Acanthamoeba-specific primers demonstrated 100% (9/9) confirmation of Acanthamoeba species. Genotyping of Acanthamoeba isolates revealed T1, T3, T4, T5, T7, T11, and T15 genotypes. Thermo-tolerance assay demonstrated that T5 and T7 genotypes were potentially pathogenic strains. The evidence of environmental distribution of Acanthamoeba spp. in the freshwater systems in the Philippines and thermo-tolerance profile of isolates are significant aspects of amoeba study in public health and calls for initiatives in the dissemination of relevant information and the expansion of knowledge, awareness, and policies on pathogenic waterborne amoeba to mitigate, prevent, detect, and report cases of human infections.

Evaluation of glutathione S-transferase polymorphism in Iranian patients with type 2 diabetic microangiopathy

BackgroundOverproduction of reactive oxygen species as a result of hyperglycemia in diabetes mellitus leads to microvascular complications. Glutathione S-transferases play important detoxifying roles with antioxidant potentials. This study aimed to assess whether the glutathione S-transferase M1 and T1 genotypes were associated with type 2 diabetes mellitus microangiopathic complications in the Iranian population.ResultsIn this case-control study, the frequencies of null GSTM1 and GSTT1 genotypes were 4/72 (5.56%) and 12/72 (16.67%) respectively, in uncomplicated DM group. The frequencies of null GSTM1 and GSTT1 genotype in complicated DM group were 16/134 (11.94%) and 37/134 (27.61%), respectively. The proportion of GSTM1 null genotypes was higher in diabetic nephropathy compared to non-nephropathy (19.3% vs. 6.04 %, P = 0.006). At GSTT1 locus, patients with diabetic peripheral neuropathy had a higher frequency of deletion compared to those of without neuropathy (30.39% vs. 23.49%) (P = 0.02).ConclusionSelective polymorphisms encoding GSTM1 and GSTT1genes may prove useful as genetic markers to recognize individuals with an increased trend in developing diabetic nephropathy and neuropathy, respectively. This will help better identify individuals at higher risk toward microvascular complications of type 2 diabetes due to genetic susceptibility.

Association of glutathione S-transferase M1 and T1 genotypes with asthma: A meta-analysis.

Background:We performed an updated meta-analysis to clarify the relationship between glutathione S-transferase Mu and theta (GSTM1 and GSTT1, respectively) null/positive genotypes and asthma.Methods:We performed a literature search using PubMed and Web of Science databases in August 2019. Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to assess the role of GSTM1 and GSTT1 genotypes in the risk of asthma.Results:Overall, we found a significant association with asthma risk in the general population for both the GSTM1 genotype (OR = 1.21; 95% CI: 1.07–1.35; P < .001; I2 = 69.5%) and the GSTT1 genotype (OR = 1.61; 95% CI: 1.30–2.00; P < .001; I2 = 83.6%). Moreover, significant associations between both genotypes and asthma risk were also found by age stratification. Furthermore, for GSTM1 we found significant associations in populations living in Asia, Europe, and Russia, but not in Africa. Conversely, for GSTT1, we found a significantly increased risk in populations living in Asia, Europe, Africa, and Russia. In addition, a significant association was found for both genotypes with a sample size <500, but not a sample size >2000.Conclusion:Our meta-analysis provides evidence that GSTM1 and GSTT1 genotypes could be used as asthma-associated biomarkers.

Association of ADAM33 T1 Polymorphism With Subgroups of Pediatric Asthma Patients in Iran

There is strong evidence on the interaction of several genetic variations and environmental conditions in the etiology of asthma. Association of a disintegrin and metalloproteinase 33 (ADAM33) with asthma risk is not clear and shows diversity between nations and ethnicities. Several single nucleotide polymorphisms (SNP) of the ADAM33 gene are introduced and studied according to the disease onset and characteristics. The aim of our study is to determine the association of ADAM33 rs2280091 polymorphism and pediatric asthma in the Iranian population. A total of 63 asthma patients (aged 6-18) and 86 healthy controls were enrolled in our study. Asthma type, classification, and severity were defined. SNPs of the ADAM33 gene at rs2280091 (T1) were analyzed. Pulmonary function tests, total blood eosinophil count, and IgE count were also assessed. T1 genotype and allele frequencies were not associated with asthma risk in Iranian pediatric asthma. Atopic asthma subgroup and patients with normal eosinophil count showed association with ADAM33 rs2280091. Moreover, asthma patients with AG genotype showed lower pulmonary functions.

Extracellular protease profile of Acanthamoeba after prolonged axenic culture and after interaction with MDCK cells.

Free-living amoebae of the genus Acanthamoeba are causative agents of Acanthamoeba keratitis and amoebic encephalitis in humans, both of which are serious infections. The ability to produce proteases is one of the factors involved in the pathogenesis of Acanthamoeba infections. The aim of this study was to evaluate the secreted proteases of six Acanthamoeba strains from distinct genotypes (T1, T2, T4 and T11) maintained in prolonged axenic culture and following three successive passages in Madin-Darby Canine Kidney (MDCK) cells. Conditioned medium was obtained from cultures before and after interaction with the MDCK monolayers, resolved in SDS-PAGE containing gelatine, then subjected to quantitative azocasein assays. Zymography profiles varied between the strains, with the predominant proteases found to be serine-type proteases from 49 to 128 kDa. A T1 genotype strain isolated from dust showed quantitatively higher protease secretion compared to the other strains. No changes were detected in the zymography profiles of MDCK-interacted cultures compared to long-term axenic cultures. Two strains presented lower proteolytic activity post-MDCK interaction, while the remaining strains presented similar values before and after MDCK passages. In conclusion, this study confirms the predominance of serine-type protease secretion by Acanthamoeba, with distinct profiles presented by the different strains and genotypes studied. Also, interaction of trophozoites with MDCK cells did not alter the zymography pattern.

The Role Of Adiponectin, TNF-α And Glutathione In The Pathogenesis And Evolution Of Type 1 Diabetes.

IntroductionType 1 diabetes (T1DM) is a chronic autoimmune or idiopathic condition, featuring complex and unique interactions between proteins and enzyme systems. The purpose of the present study is to investigate the role of AdipoQ +276G>T, TNF-α-308G>A, GSTT1/GSTM1 polymorphic variants in the development of T1DM.Materials and methodsThe study is designed as a cross-sectional study, involving 72 diabetic cases and 90 controls. Genotyping was carried out according to specific protocols for the above-mentioned polymorphic variants.ResultsThe G allele of AdipoQ was associated with the development of type 1 diabetes (OR 0.577, CI95% 0.336–0.802, p=0.001), similar to the GG and GA genotypes (OR 0.405, CI95% 0.156–0.654, p=0.001 and OR 0.623, CI95% 0.401–0.855, p=0.004). The G allele of TNF-α was marginally associated with the development of type 1 diabetes (OR 0.789, CI95% 0.579–0.956, p=0.005). The presence of the T1 genotype was a strong predictor for type 1 diabetes (OR 3.4, CI95% 1.433–6.243, p<0.001).ConclusionThe results of our study suggest that G alleles of AdipoQ and TNFα act as a protective factor in T1DM, while the T1 allele for GST could be considered a risk factor for the development of Type 1 diabetes in our study group.

Prevalence of glutathione s-transferase genes some polymorphisms in menopausal women of two ethnic groups with insomnia

Introduction: According to the current data, most menopausal women have reduced quality of life and one of the main criteria of it is sleep disturbances. It is highly possible, that main consequences of sleep loss may be oxidative stress. Glutathione system is the one of the important components of the antioxidant defense system, which includes glutathione and enzymes. It has been shown that the activity of these enzymes is determined by the polymorphic variants of the corresponding genes. The aim of this research is to carry out a comparative analysis of the frequency distribution of Glutathione S-transferase T1 (GSTT1) and M1 (GSTM1) genotypes in Caucasian and Asian menopausal women with insomnia.

The Relationship Between Polymorphisms of Glutathione S-Transferase (GSTM1, GSTT1) Gene and Type 2 Diabetes Mellitus in Western Iran?

Background: Diabetes is a chronic metabolic disorder that causes many complications in the body. Oxidative free radicals affect human beta cells. Glutathione S-transferases (GSTs) can deactivate substances formed in oxidative stress. Objectives: The present study was conducted to examine the relationship between GSTs polymorphisms and type 2 diabetes mellitus (T2DM), and to investigate the presence of GSTM1 and T1 genotypes in 176 T2DM patients and 174 healthy cases as controls. Methods: Multiplex PCR was used to confirm the presence or absence of GSTT1 and M1 genotypes. Conclusions: The obtained results indicated that the polymorphism of GSTM1 and GSTT1 does not affect the prevalence of T2DM in the study population.

Association of ADAM33 polymorphisms to the presence, severity and control of asthma in a Brazilian sample population

Asthma is an inherited disease that results in complex inflammation of the lungs, obstruction of the airways and bronchial hyperresponsiveness. The analysis of genes that act in the Asthma is recent, and one of these genes as well as its variants most studied in asthma in different populations is ADAM33, responsible for the synthesis of a homonymous glycoprotein that acts restricted in cellular processes of the smooth muscles of the airways. The objective of the study was to verify the genotypic frequency of rs2280091 (T1) and rs2280090 (T2) variants in ADAM33 gene, and analyze whether they act in the presence, control and severity of asthma. The analysis was performed using the PCR-RFLP technique of 51 patients with asthma and 109 healthy individuals. In agreement with some other studies in different populations, our results showed no association between frequency of genotype of T1 and asthma. However, we noticed a positive association between T2 genotype and asthma.

Molecular characterization of Acanthamoeba strains isolated from the oral cavity of hemodialysis patients in Iran.

Free-living amoebae (FLA) of the genus Acanthamoeba are opportunistic pathogenic agents able to cause life-threatening infections in immunosuppressed patients. Chronic kidney disease impairs adaptive and innate immunity. Thus, patients with chronic kidney disease are prone to opportunistic infections by potentially pathogenic FLA. Therefore, in the present study, the investigation of Acanthamoeba genotypes isolated from the oral cavity of hemodialysis patients of reference hospitals in Iran was aimed, using both morphology and molecular (sequence-based analysis) tools. Furthermore, classification of the strains at the genotype level was performed on the basis of differences in the diagnostic fraction 3 (DF3) region of the 18S rRNA gene. The pathogenic potential of the isolated amoebae was also determined using thermotolerance and osmotolerance assays. Out of the 187 oral cavity samples investigated, nine (4.8%) were positive for FLA. DNA sequencing of the ASA.A1 region of the 18S rRNA gene revealed that the isolated strains belonged to the Acanthamoeba T1 and T4 genotypes. Genotype T1 was isolated for the first time from a patient in Iran. Interestingly, the T1 strain (AN2 strain) exhibits a high pathogenic potential in tolerance assays. The pathogenicity assay revealed that five strains were able to grow at high temperatures (37-40°C) and high osmolarity (0.5 and 1M D-mannitol) conditions; thus, they were considered as potentially pathogenic strains. Moreover, two of the patients were positive for Vermamoeba genus. The present study is the first report of genotype T1 isolation in Iran and the first to identify the occurrence of Acanthamoeba and Vermamoeba genera in patients undergoing hemodialysis worldwide. Monitoring hemodialysis and renal failure patients should be a priority for possible control of Acanthamoeba and other FLA-related diseases.

Molecular characterization of emerging G9P[4] rotavirus strains possessing a rare E6 NSP4 or T1 NSP3 genotype on a genogroup-2 backbone using a refined classification framework.

Rotavirus infections associated with unusual strains are an emerging concern in rotavirus vaccination programmes. Recently, an increase in circulation of unusual G9P[4] strains was reported from different regions of India, placing this genotype in third position, after G1P[8] and G2P[4], of the most common rotavirus strains. The aim of the present study was to analyse the complete genomic constellation of three G9P[4] strains (RV09, RV10 and RV11), determine their genetic relatedness to other genogroup-2 strains and understand the evolution of a rare E6 and other NSP4 genotypes. All strains revealed the presence of a genogroup-2 backbone, with RV09 constituting the NSP3 T1 genotype and RV10 and RV11 bearing the NSP4 E6 genotype. A refined criterion adopted to classify the nine internal gene segments of G2P[4] and non-G2P[4] strains with the genogroup-2 backbone into lineages and sub-lineages indicated divergence of >8 % (except NSP1: >5.5 %) for lineages and >3 % for sub-lineages. The VP1 and/or VP3 genes of study strains showed close relationships with animal-like human rotaviruses. The estimated evolutionary rate for the NSP4 E6 genotype was marginally higher (3.78×10-3 substitutions per site per year) than that of genotypes E1 (2.6×10-3 substitutions per site per year) and E2 (3.06×10-3 substitutions per site per year), suggesting a step towards adaptation of E6 on a genogroup-2 backbone. The time and origin of the most recent common ancestor of E6 genotype were estimated to be 1981 and South Asia, respectively. Full-genome and evolutionary analyses performed in this study for G9P[4] strains will help better understand the extent of gene reassortment and origin in unusual rotavirus strains that may remain viable and cause infections in humans.

Association between polymorphism of ADAM33 gene and bronchial asthma in Mongolian population

Objective: To investigate the different genotype and allele frequency distribution of ADAM33 gene T1, T2, V4, S2 sites Mongolian population, and discuss the relationship between ADAM33 gene polymorphism and bronchial asthma. Methods: From January 2014 to December 2015, a total of 180 cases of Mongolian patients with asthma were detected, compared with 186 cases of healthy Mongolian as controled and screening significant genes.Selected restriction fragment length polymorphism (PCR-RFLP) method to detected ADAM33 gene polymorphism.According to condition , the asthma group was divided into mild(n=83), medium group(n=47)and severe group(n=50). The distribution difference of different genotype and every genotype of V4 FEV1, eosinophils, IgE comparison were compared , and analysis their correlation. Results: In ADAM33 , the distribution of T1 sites (AA and AG genotypes) had statistical significance compared asthma group with control group(χ2=8.810, 8.294, P<0.05, OR=1.983, 0.500). The OR value of G allele was 0.580.The distribution of S2 site(CC genotype) had statistical significance(χ2=4.277, P<0.05), the OR value of G allele was 1.423.the distribution of V4 sites (GC and GG genotypes) had statistical significance between the two groups (χ2=7.880, 10.313, P<0.05), OR value was 0.459, 2.130, G allele OR value was 1.496.The distribution frequency difference of each genotype in V4 sites in mild, medium and severe group was statistically significant (χ2=16.049, P<0.05), and compared various genotypes of FEV1, IgE, the difference was statistically significant (P<0.05), for each genotype of T2 site in asthma group and the control group there was no statistically significant in the distribution (χ2=1.218, 0.248, 1.287, P>0.05). Conclusions: T1, V4, S2 locus polymorphism of ADAM33 gene may play a role in the Mongolian asthma population, and T2 locus polymorphism may has no relationship with Mongolian asthma patients.And the genotype polymorphism of V4 sites may be associated with asthma severity.