Genotype Groups Research Articles

Abstract P3066: TSC2-mTOR Axis Exerts Biased Control Over Macrophage Infiltration Following Myocardial Infarction

Introduction: The mechanistic target of rapamycin complex 1 (mTORC1) is an evolutionarily conserved serine/threonine kinase, which regulates pivotal physiological processes. mTORC1 activity is tightly regulated upstream by the tuberous sclerosis complex (TSC) which consists of TSC1 and TSC2. In macrophages as throughout the immune system, mTORC1 is a potent regulator of cell activation and function. Myocardial infarction (MI) is associated with stimulation of mTORC1; however, studies vary as to whether this activation is beneficial or detrimental, and its influence on macrophage differentiation, and infiltration after MI is unknown. Hypothesis: We tested the hypothesis that cardiac infiltration of macrophages following MI is regulated by their expression of TSC2. Methods/Results: Macrophage-specific TSC2 knock-out mice (TSC2-LyzM) were made by crossing mice with floxed-TSC2 and LysM-Cre that display a significant reduction of macrophage TSC2 protein. TSC2-LyzM bone marrow-derived macrophages (BMDMs) have constitutive mTORC1 activation reflected by phosphorylation of downstream targets S6K1 and 4E-BP1 and were correspondingly larger. To assess macrophage polarization, cells were stimulated with LPS to promote an M1-like phenotype or IL-4 to induce an M2-like phenotype. LPS- treated TSC2-LyzM BMDMs secreted more of the proinflammatory cytokine TNF-a, while IL-4 stimulation resulted in less of an M2 program, with significantly less Arg1, Mrc1, and Rentla expression. These findings show TSC2 is required for normal BMDM polarization. We next tested if TSC2 impacts macrophage myocardial infiltration after MI. Macrophages were identified as CD45+CD11b+CD64+ and further parsed by CCR2 and MHC-II. In concordance with prior studies, we found substantial infiltration of CCR2+ (BMDMs) macrophages in both genotype groups after MI. However, more TSC2-LyzM had increased CCR2+ MHC-II+ phenotype (infiltrating macrophages) versus wild-type. Conclusion: Overall, our findings highlight a key role for the TSC2-mTORC1 pathway in regulating macrophage fate specification and cardiac infiltration following a MI. This supports novel signaling control over BMDMs by TSC2-phosphorylation mediated mTORC1 activation or inhibition.

Abstract P2076: Disrupting Nicotinamide Nucleotide Transhydrogenase Prevents Cardiometabolic Heart Failure With Preserved Ejection In Mice.

Heart failure with preserved ejection fraction (HFpEF) represents a common clinical endpoint of cardiometabolic diseases which impair myocardial diastolic relaxation and/or stiffness. Although myocardial redox perturbations are known to accompany HFpEF, their contribution to its pathogenesis has not been clearly demonstrated. Therefore, our multi-cohort murine study sought to determine whether constitutive expression of NNT, a mitochondrial transhydrogenase that couples NADPH:NADP + to NADH:NAD + homeostasis, is required for the cardiometabolic manifestations of HFpEF in mice. Four cohorts of 12-week-old male and female mice with wild-type (NNT +/+ ) or mutant (NNT -/- ) NNT were supplied a combined high-fat and 0.5% N(ω)-nitro-L-arginine methyl ester (HFD-LNAME) diet for 9 weeks (n = 6-10). Although male NNT +/+ mice developed expected increases in body weight (23.2% Δ, P = 0.003), hypertension (24 ± 5 Δ mmHg, P = 0.02), impaired glucose tolerance ( P = 0.006), and reduced maximal treadmill running distance (-172 ± 73.1 Δ m, P = 0.006) following 9 weeks HFD-LNAME, male NNT -/- mice were protected against such cardiometabolic dysfunction. As previously shown, female mice failed to develop cardiometabolic dysfunction independent of NNT genotype. Cardiac phenotyping via echocardiography and invasive PV-loop analysis revealed increased E/e’ (42.8 vs. 21.5, P < 0.001) and E/A (2.3 vs 1.4, P = 0.007) ratios, diastolic stiffness (0.09 vs 0.04 mmHg/μL, P = 0.02), and myocardial fibrosis ( P = 0.02). Myocardial transcriptomic analysis via RNA-sequencing revealed robust unsupervised clustering of both genotypic and dietary groups, which ultimately showed disproportionate perturbations mitochondrial metabolic and redox signaling intermediate genes. Our search for putative transcriptional regulators identified NAD + deacetylase Sirt3 as an NNT-dependent oxidative response gene that appears to underlie cardiometabolic HFpEF. Taken together, these observations support that the loss of NNT protects against nitrosative stress-induced cardiac and metabolic consequences of HFD-LNAME in mice, thus highlighting a novel therapeutic avenue for HFpEF.

Survey of Xylella fastidiosa Infection in Prunus Germplasm in Gainesville, FL, USA

The pathogen Xylella fastidiosa is a xylem-restricted, gram-negative bacterium that is known to cause diseases of many cultivated plant species. Recent outbreaks of X. fastidiosa diseases in Europe have brought attention to the impact of this pathogen, especially to perennial crops. Among the Prunus genus, X. fastidiosa is known to have a wide range of hosts, including plum, almond, peach, cherry, and apricot. Infected trees have reduced fruit quality, possibly resulting in unmarketable fruits, as well as reduced lifespan. There are no curative management options for X. fastidiosa diseases in Prunus; therefore, development of resistant or tolerant cultivars through breeding represents an efficient option to reduce the impact of this pathogen. In this context, the main objective of this study was to determine the occurrence of X. fastidiosa in germplasm of the Stone Fruit Breeding Program at the University of Florida located in Gainesville, FL, USA, under natural infection conditions. A total of 43 individuals representing 10 different genotypic groups within the Prunus genus were tested for the presence of X. fastidiosa. Additionally, we report a novel and easy sampling method using sawdust collected from tree trunks for the detection of this pathogen in Prunus and the development of an endogenous control for improving the diagnosis of this pathogen using real-time polymerase chain reactions. Our results showed a high incidence of X. fastidiosa in the germplasm tested, with more than 65% of the samples positive for the presence of the bacterial pathogen. However, X. fastidiosa was not detected in most of the P. mume samples tested, whereas almost all the P. mume × P. armeniaca hybrids were positive. Negative individuals were also identified in P. avium, P. campanulata, P. umbellata, and P. salicina × P. ceracifera. These trees have been planted in the field, exposed to natural infection for 4 to 11 years, and are considered to show field resistance. Finally, primers and probes based on the Prunus COX gene developed in this study can be used as an internal amplification control to enhance the interpretation of results of X. fastidiosa detection assays using sawdust samples.

Narrow root cone angle promotes deeper rooting, cooler canopy temperatures and higher grain yield in a rice (Oryza sativa L.) recombinant inbred line population grown under different water availabilities in aerobic production systems

Rice crops are one of the world’s major consumers of irrigation water. With water becoming more limited, water-saving aerobic production (AP) systems may provide an alternative to traditional flooded culture. Genotypes with narrow root cone angle (RCA) and subsequently deeper rooting systems are hypothesised as key for AP adaptation ensuring water uptake at depth. Using a recombinant inbred line (RIL) population and differential genotypes (narrow and wide RCA groups) derived from a Sherpa/IRAT109 cross, a glasshouse experiment and four field experiments with different water inputs in well-watered (WW, in two seasons), mild water deficit (MWD), and intermittent water deficit (IWD) conditions were carried out to characterise the relationship of RCA with grain yield (GY) and key physiological traits. Significant genotypic variation and group differences were found in root traits, canopy temperature and GY. Expression of genotypes with narrow RCA were found to be consistent across experiments and water availability conditions. Genotypes in the narrow RCA group produced a greater proportion of roots below 20 cm (7.4–17.7%), longer total root length (69.7%), cooler canopies (0.3–1.0 °C) and higher GY (14–62%) compared with wide RCA group. Two promising RILs were identified which expressed high GY in WW (12.0–13.4 t/ha) conditions while maintaining GY in IWD (up to 9.4 t/ha). Higher GY was obtained in genotypes with cooler canopies (r = −0.72) and narrow RCA (r = −0.39) in IWD. Narrower RCA was associated with higher total root length (r = −0.70) and higher stomatal conductance (r = −0.71) in the glasshouse and proportion of deep roots (r = −0.65) and cooler canopies (r = 0.66) in the field. To our knowledge, this was one of the foremost studies exploring and demonstrating such relationships in rice AP systems. Root traits determined in the field were congruent with the other root-related traits driving differences between RCA groups measured in the glasshouse providing confidence with the field phenotyping methods. This work demonstrated the advantage conferred by the development of narrow RCA in aerobic conditions, presented improved physiological understanding of RCA and with genomics assisted breeding, is considered valuable for the development of AP-adapted, sustainably produced rice.

Association Analysis between Genotype and Phenotype of α,β-Thalassaemia Carriers in Huizhou Area of Guangdong Province

To analyze the prevalence, genotype distribution and hematological characteristics of α,β-thalassaemia carriers in Huizhou area of Guangdong Province. 10 809 carriers of simple β-thalassaemia and 1 757 carriers of α,β-thalassaemia were enrolled as our study cohort. The hematological parameters were detected by automated blood cell counters and automatic capillary electrophoresis. Suspension array technology, gap-polymerase chain reaction (gap-PCR) and PCR-reverse dot blot were used for the genotyping of thalassaemia carriers. The prevalence of α,β-thalassaemia in Huizhou area of Guangdong Province was 1.99%. A total of 62 genotypes were detected, and the most prevalent genotype was --SEA/ αα, βCD41-42/ βN (19.29%), the next was --SEA/ αα, βIVS-II-654/ βN (16.73%). Significant differences in mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) were found between different genotype groups for simple β-thalassaemia and α,β-thalassaemia. Violin plots showed that carriers with co-inheritance of β-thalassaemia and mild α-thalassaemia expressed the lightest anemia, and carriers with co-inheritance of β-thalassaemia and hemoglobin H (Hb H) disease expressed the most severe anemia. There is a high prevalence of α,β-thalassaemia in Huizhou area of Guangdong Province. Because of the lack of specific hematological makers for diagnosis of α,β-thalassaemia, it is necessary to distinguish it from simple β-thalassaemia by genotyping of α- and β-thalassaemia in order to correctly guide genetic counseling and prenatal disgnosis.

Association between LEPR Genotype and Gut Microbiome in Healthy Non-Obese Korean Adults.

The LEPR (leptin receptor) genotype is associated with obesity. Gut microbiome composition differs between obese and non-obese adults. However, the impact of LEPR genotype on gut microbiome composition in humans has not yet been studied. In this study, the association between LEPR single nucleotide polymorphism (rs1173100, rs1137101, and rs790419) and the gut microbiome composition in 65 non-obese Korean adults was investigated. Leptin, triglyceride, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol levels were also measured in all participants. Mean ± SD (standard deviation) of age, body mass index, and leptin hormone levels of participants was 35.2 ± 8.1 years, 21.4 ± 1.8 kg/m2, and 7989.1 ± 6687.4 pg/mL, respectively. Gut microbiome analysis was performed at the phylum level by 16S rRNA sequencing. Among the 11 phyla detected, only one showed significantly different relative abundances between LEPR genotypes. The relative abundance of Candidatus Saccharibacteria was higher in the G/A genotype group than in the G/G genotype group for the rs1137101 single nucleotide polymorphism (p=0.0322). Participant characteristics, including body mass index, leptin levels, and other lipid levels, were similar between the rs1137101 G/G and G/A genotypes. In addition, the relative abundances of Fusobacteria and Tenericutes showed significant positive relationship with plasma leptin concentrations (p=0.0036 and p=0.0000, respectively). In conclusion, LEPR genotype and gut microbiome may be associated even in normal-weight Korean adults. However, further studies with a greater number of obese adults are needed to confirm whether LEPR genotype is related to gut microbiome composition.

Bioactive Compounds and Antioxidant Activity in Seeds of Bred Lines of Common Bean Developed from Interspecific Crosses.

Knowledge is limited about the level of bioactive compounds and antioxidant activity of seeds from bred lines of common beans developed from interspecific crosses using four different Phaseolus species (P. vulgaris L., P. coccineus L., P. acutifolius A. Gray. Gray., and P. dumosus). In this study, differences in the nutritional quality of seeds among 112 bean genotypes were evaluated by measuring the levels of phenolic compounds, pigments, antioxidant activity, and sugars. The bean genotypes were grown under high temperatures and acid soil conditions in the Amazon region of Colombia. Five typology groups of bean genotypes were identified based on the level of bioactive compounds and their functional capacity: (1) highly bioactive and functional (HBF); (2) moderately bioactive and functional (MBF); (3) moderate antioxidant content with pigment influence (MACP); (4) moderately antinutritional with limited antioxidant potential (MALAP); and (5) antinutritional, low bioactive, and functional (ALBF). We developed a nutritional quality index (NQI) with values ranging from 0 to 1 based on the nutritional and anti-nutritional balance of each genotype and the higher values of the NQI of a genotype indicating greater nutritional quality. We found three interspecific bred lines (SER 212, SER 213, and RRA 81), with NQI values higher than 0.8. These three lines belong to the typology group of HBF. The superior nutritional quality of these three interspecific bred lines is attributed to a greater level of bioactive compounds and antioxidant capacity. These three bred lines may serve as useful parents to develop nutritionally superior and stress-resilient beans from bean breeding programs. Further research is needed to explore the role of testa color in improving the nutritional quality of seeds of common bean genotypes grown under different climatic conditions.

Evaluation of inactivated avian influenza virus and Newcastle disease virus bivalent vaccination program against newly circulated H5N8 and NDV strains

Avian influenza virus (AIV) and Newcastle disease virus (NDV) are respiratory illness syndromes that have recently been detected in vaccinated flocks and are causing major financial losses in the chicken farming industry. The objective was to evaluate the efficacy of Valley Vac H5Plus NDVg7 vaccine in protecting chickens against the H5N8 and NDV strains that have recently been circulating in comparison with the efficacy of the commercially available bivalent H5+ND7 vaccine. In contrast to the H5+ND7 vaccine, which was made of genetically distinct H5N8/2018 clade 2.3.4.4b genotype group (G5), H9N2/2016, H5N1/2017, and genetically comparable NDV genotype VII 1.1/2019 of the recently circulating challenge viruses, the Valley Vac H5Plus NDVg7 vaccine consisted of the recently isolated (RG HPAI H5N1 AIV/2015 Clade 2.2.1.2, RG HPAIV H5N8/2020 Clade 2.3.4.4b genotype group 6 (G6), and NDV genotype VII 1.1/2012) which were genetically similar to challenged strains. To determine the effectiveness of the Valley Vac H5Plus NDVg7 vaccine, a total of 70-day-old commercial chicks were divided into 7 groups of 10 birds each. Groups (G1 and G4) received Valley Vac H5Plus NDVg7 vaccine. Groups (G2 and G5) groups received commercial H5+ND7 vaccine. While groups (G3 and G6) were kept nonvaccinated, and group (G7) was kept as a nonchallenged and nonvaccinated. After 3-wk post vaccination (WPV), groups G1, G2, and G3 were challenged with A/Duck/ Egypt/SMG4/2019(H5N8) genotype G6. On the other hand, groups G4, G5, G6 were challenged with NDV/EGYPT/18629F/2018 genotype VII 1.1 with an intranasal injection of 0.1 mL. Antibody titer was calculated at the first 3 wk after vaccination, and the viral shedding titer was calculated at 3-, 5-, and 7-days post challenge. Mortality and morbidity rates were monitored daily during the experiment, and for the first 10 d after the challenge, to provide an estimate of the protection rate. The results showed that a single dosage of 0.5 mL per bird of Valley Vac H5Plus NDVg7 vaccine provides 80% protection against both H5N8 and NDV, compared to the bivalent H5+ND7 vaccine, which provided 20 and 80% protection against H5N8 and NDV, respectively. In addition, 0.5 mL per bird of Valley Vac H5Plus NDVg7 vaccine produced a greater immune response against both viruses than commercial vaccination at 1 to 3 WPV with a significant difference at 1 WPV for H5N8 and a comparatively higher immune response for NDV. Furthermore, it reduced virus shedding of H5N8 on the third, fifth, seventh, and tenth days lower than H5+ND7 vaccine with a significant difference on the third day for H5N8 and relatively lower than bivalent H5+ND7 vaccine for NDV with a significant difference on the fifth day. The Valley vaccinated group demonstrated more tissue intactness compared to the commercially vaccinated group against the H5N8 challenge, however the bivalent commercially vaccinated group showed the similar level of tissue integrity against NDV. In conclusion, Valley Vac H5Plus NDVg7 that contains the genetically similar strain to recently circulating challenged virus (H5N8 genotype G6) provided better protection with greater immune response and decreased the amount of virus shed against H5N8 genotype G6 and showed less histopathological alteration than the commercial bivalent H5+ND7 vaccine that contain genetically distinct (H5N8 genotype G5). However the Valley Vac H5Plus NDVg7 provided the same protection with relatively high immune response and relatively decreased the amount of virus shed and showed equal tissue integrity than the commercial bivalent H5+ND7 vaccine against NDV genotype VII 1.1 that contain the same genotype of NDV genotype VII 1.1.

Transcriptome analysis reveals key regulatory genes for root growth related to potassium utilization efficiency in rapeseed (Brassica napus L.).

Root system architecture (RSA) is the primary predictor of nutrient intake and significantly influences potassium utilization efficiency (KUE). Uncertainty persists regarding the genetic factors governing root growth in rapeseed. The root transcriptome analysis reveals the genetic basis driving crop root growth. In this study, RNA-seq was used to profile the overall transcriptome in the root tissue of 20 Brassica napus accessions with high and low KUE. 71,437 genes in the roots displayed variable expression profiles between the two contrasting genotype groups. The 212 genes that had varied expression levels between the high and low KUE lines were found using a pairwise comparison approach. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional classification analysis revealed that the DEGs implicated in hormone and signaling pathways, as well as glucose, lipid, and amino acid metabolism, were all differently regulated in the rapeseed root system. Additionally, we discovered 33 transcription factors (TFs) that control root development were differentially expressed. By combining differential expression analysis, weighted gene co-expression network analysis (WGCNA), and recent genome-wide association study (GWAS) results, four candidate genes were identified as essential hub genes. These potential genes were located fewer than 100 kb from the peak SNPs of QTL clusters, and it was hypothesized that they regulated the formation of the root system. Three of the four hub genes' homologs-BnaC04G0560400ZS, BnaC04G0560400ZS, and BnaA03G0073500ZS-have been shown to control root development in earlier research. The information produced by our transcriptome profiling could be useful in revealing the molecular processes involved in the growth of rapeseed roots in response to KUE.

Correlation of N-acetyltransferase 2 genotype and acetylation status with plasma isoniazid concentration and its metabolic ratio in ethiopian tuberculosis patients

Unfavorable treatment outcomes for tuberculosis (TB) treatment might result from altered plasma exposure to antitubercular drugs in TB patients. The present study investigated the distribution of the N-Acetyltransferase 2 (NAT2) genotype, isoniazid acetylation status, genotype–phenotype concordance of NAT2, and isoniazid plasma exposure among Ethiopian tuberculosis patients. Blood samples were collected from newly diagnosed TB patients receiving a fixed dose combination of first-line antitubercular drugs daily. Genotyping of NAT2 was done using TaqMan drug metabolism assay. Isoniazid and its metabolite concentration were determined using validated liquid chromatography-tandem mass spectrometry (LC–MS/MS). A total of 120 patients (63 male and 57 female) were enrolled in this study. The mean daily dose of isoniazid was 4.71 mg/kg. The frequency of slow, intermediate, and fast NAT2 acetylators genotypes were 74.2%, 22.4%, and 3.3% respectively. The overall median isoniazid maximum plasma concentration (Cmax) was 4.77 µg/mL and the AUC0–7 h was 11.21 µg.h/mL. The median Cmax in slow, intermediate, and fast acetylators were 5.65, 3.44, and 2.47 μg/mL, respectively. The median AUC0–7 h hour in slow, intermediate, and fast acetylators were 13.1, 6.086, and 3.73 mg•h/L, respectively. The majority (87.5%) of the study participants achieved isoniazid Cmax of above 3 µg/mL, which is considered a lower limit for a favorable treatment outcome. There is 85% concordance between the NAT2 genotype and acetylation phenotypes. NAT2 genotype, female sex, and dose were independent predictors of Cmax and AUC0–7 h (p < 0.001). Our finding revealed that there is a high frequency of slow NAT2 genotypes. The plasma Cmax of isoniazid was higher in the female and slow acetylators genotype group. The overall target plasma isoniazid concentrations in Ethiopian tuberculosis patients were achieved in the majority of the patients. Therefore, it is important to monitor adverse drug reactions and the use of a higher dose of isoniazid should be closely monitored.

High blood levels of brain-derived neurotrophic factor (BDNF) mRNA in early psychosis are associated with inflammatory markers

The brain-derived neurotrophic factor (BDNF) single nucleotide polymorphism (SNP) rs6265C > T, Val66Met, affects BDNF secretion and has been related to inflammatory processes. Both the rs6265 and BDNF protein levels have been widely investigated in neuropsychiatric disorders with conflicting results. In the present study we examined BDNF mRNA expression in blood considering the SNP rs6265 and its relationship with inflammatory markers in the early stages of psychosis. The rs6265 genotype and blood BDNF mRNA levels were measured in 34 at-risk mental states (ARMS) individuals, 37 patients with first-episode psychosis (FEP) and 42 healthy controls (HCs) by quantitative PCR and reverse transcription (RT)-qPCR using validated TaqMan assays. We also obtained measures of interleukin-6 (IL6) mRNA levels, fibrinogen, neutrophil-to-lymphocyte ratio (NLR) and high-sensitivity C-reactive protein. We identified that BDNF mRNA levels were associated with the rs6265 genotype in an allele-dose-dependent manner, with low expression levels associated with the T allele (Met substitution). Thus, we controlled for the rs6265 genotype in all analyses. Blood BDNF mRNA levels differed between diagnostic groups: patients with FEP exhibited higher blood BDNF mRNA levels than ARMS individuals, and the lowest levels were observed in HC. In addition, we observed significant correlations between BDNF mRNA levels and inflammatory markers (IL6 mRNA levels and NLR), controlled by the rs6265 genotype, in ARMS and FEP groups. This exploratory study suggests that the rs6265 genotype is associated with differential blood mRNA expression of BDNF that increases with illness progression and correlated with inflammation in the early stages of psychosis.

Genetic diversity analysis in sunflower (Helianthus annuus L.) restorer lines using SSR markers

Sunflower is a staple oilseed crop ofthe world. Genetic diversity in the parental lines is key for hybrid breeding programmes in sunflower. In this study, genetic diversity in a set of 102 parental lines (100 restorer and twoCMS) of sunflower was assessed using 69 polymorphic SSR markers. The genetic diversity parameters: average number of alleles (3.07) per locus, gene diversity (0.356) and polymorphism information content (0.296) revealed low to moderate genetic diversity in the restorer lines. The SSR marker ORS447 located on linkage group 2 was more informative with high number of alleles (10) and high PIC value (0.89). Cluster analysis (neighbour-joining tree) revealed three major genotypic groups. Model based STRUCTURE analysis showed recognizable population structure; based on membership coefficients(&gt;80%), 82 genotypes were classified into two populations (K=2) and the remaining 20 genotypes were classified into admixture group. The Fst value (0.278) suggested that the populations were differentiated. Analysis of molecular variance resultsshowed that maximum of genetic variation (72%) was observed between the individuals within the population suggesting that the population was weakly structured. These results would be useful for selecting SSR markers for genotype characterization as well as choosing diverse parents for hybrid development programme in sunflower.

Detection of Transversions and Transitions in HBG2 Cis-Elements Associated with Sickle Cell Allele in Ghanaians.

Short tandem repeats located 5' prime to the β-globin gene, have been observed to be in linkage disequilibrium with the HbS allele, and thought to affect the severity of sickle cell disease. Here, we report on new mutants within the HBG2 region that may impact sickle cell disease. To determine the cis-acting elements microsatellites, indels and single nucleotide polymorphisms (SNPs), within the HBG2 region by sequencing, in subjects with sickle cell disease. The case-control study was located at the Center for Clinical Genetics, Sickle cell unit, Korle-Bu Teaching Hospital. A questionnaire was used for demographic data and clinical information. Hematological profile (red blood cell, white blood cell, platelet, hemoglobin and mean corpuscular volume) were assessed in 83 subjects. A set of 45 samples comprising amplified DNA on the HBG2 gene from HbSS (22), HbSC (17) and 6 controls (HbAA) were sequenced. Differences in the microsatellite region between sickle cell disease (SCD) (HbSS and HbSC) genotypes and control subjects were identified by counting and assessed by Chi-square analysis. Red blood cells, hematocrit, platelets, white blood cells and hemoglobin indices differed in genotypic groups. HbSS subjects were affirmed to have severer hemolytic anemia than HbSC subjects. Two indels (T1824 and C905) were seen in both SS and SC genotypes. Two peculiar SNPs: G:T1860 (transition) and A:G1872 transversions were found within the HBG2 gene that were significantly associated with the HbSS genotype (Fisher's exact test, p = 0.006) and HbS allele respectively (Fisher's exact test, p = 0.006). Cis-acting elements in HbSS and HbSC were different and may contribute to the phenotype seen in the disease state.

ADH1B, ADH1B/C and CYP2E1 Gene Polymorphism and the Risk of Fetal Alcohol Spectrum Disorder.

Increasing alcohol consumption by women of childbearing age contributes to more frequent cases of fetal alcohol spectrum disorder. The cause of the syndrome is fetal alcohol exposure, particularly what is referred to as high prenatal alcohol exposure. Low metabolic activity of fetal enzymes shifts the burden of ethanol removal to maternal metabolism. One of the factors influencing the pathogenesis of FASD is the genetic background. It can determine the rate of elimination of ethanol, thus increasing or decreasing the time of fetal exposure to ethanol and also decreasing its concentration. Genetic polymorphisms could potentially play a significant role in these processes. In the present study, we considered three polymorphisms of genes implicated in the synthesis of enzymes involved in ethanol metabolism, i.e., ADH1b (rs1229984), ADH1b/c (rs1789891), and CYP2E1 (rs3813867). The studied group consisted of 303 children and 251 mothers. Both mothers' and children's genotypes were considered in our analysis. There were no statistically significant differences between the respective groups of genotypes of the studied polymorphisms. However, the genetic background of FASD is still elusive.

Investigation of Genetic Diversity and Grouping of Barley Genotypes Based on Indices Related to Grain Yield under Rain-fed and Supplemental Irrigation Conditions

Investigation of Genetic Diversity and Grouping of Barley Genotypes Based on Indices Related to Grain Yield under Rain-fed and Supplemental Irrigation Conditions

Morpho-physiological traits and SSR markers-based analysis of relationships and genetic diversity among fodder maize landraces in India.

Maize is an excellent fodder crop due to its high biomass, better palatability, succulency, and nutrition. Studies on morpho-physiological and biochemical characterization of fodder maize are limited. The present study aimed to explore the genetic variation in fodder maize landraces for various morpho-physiological traits and estimation of genetic relationship and population structure. The study on 47 fodder maize landraces revealed significant variation for all morpho-physiological traits except leaf-stem ratio. Plant height, stem girth, leaf-width and number of leaves showed positive correlation with green fodder yield. Morpho-physiological traits-based clustering grouped the landraces into three major clusters, whereas neighbour joining cluster and population structure analysis using 40 SSR markers revealed four and five major groups, respectively. Most landraces of Northern Himalaya-Kashmir and Ludhiana fall into a single group, whereas rest groups mainly had landraces from North-Eastern Himalaya. A total of 101 alleles were generated with mean polymorphic information content value of 0.36 and major allele frequency of 0.68. The pair wise genetic dissimilarity between genotypes ranged from 0.21 to 0.67. Mantel test revealed weak but significant correlation between morphological and molecular distance. Biochemical characterisation of superior landraces revealed significant variation for neutral detergent fibre, acid detergent fibre, cellulose and lignin content. Interestingly, significant, and positive correlation of SPAD with lignin content can be explored to bypass the costly affair of invitro quality assessment for digestibility parameters. The study identified superior landraces and demonstrated the use of molecular markers in genetic diversity assessment and grouping of genotypes for fodder maize improvement.

First study of genital HPV infection among women living with HIV recruited from May to September 2018 in Eastern Algeria.

Genital HPV infection is recognized worldwide as one of the most common, if not the most common, sexually transmitted infection. Several studies have shown a higher prevalence of HPV infection in women living with HIV (WLWH). We aimed to determine the prevalence of HPV, the circulating genotypes, and its association with risk factors among Algerian WLHIV. Cervical specimens were collected from 100 WLHIV. The detection of HPV infection was performed by Roche Linear Array test. The overall prevalence of HPV infection was determined at 32%, all including high risk HPV (HR-HPV). We found a great diversity of HR-HPV genotypes, the most common individually were HPV52, HPV16, HPV18 and HPV58, The highest prevalence was found in groups of genotype 52, 25%. HPV16 and 18 were less common (16%). Cervical cytology was abnormal in 66% (81.3% in the HPV + population), with a predominance of inflammatory lesions (75% HPV +), The most important determining risk factor for HPV infection in this series was a low CD4 T cell count <200/mm3 found in 72% of the HPV positive participants. Our study is an initial database, to be completed by a multicenter study to determine the most frequent genotypes in Algerian WLHIV, in order to discuss the introduction of the vaccine against HPV in Algeria, especially among WLHIV.

Assessing the in vivo efficacy of rational antibiotics and combinations against difficult-to-treat Pseudomonas aeruginosa producing GES β-lactamases.

We evaluated the in vivo efficacy of human-simulated regimens (HSRs) of cefiderocol, ceftazidime/avibactam, meropenem and ceftazidime/avibactam/meropenem combination against Guiana-extended spectrum (GES)-producing Pseudomonas aeruginosa isolates. Eighteen P. aeruginosa isolates producing GES-1 (n = 5), GES-5 (n = 5) or miscellaneous GESs (combinations of GES-19, GES-20 and/or GES-26; n = 8) were evaluated. In vitro MIC testing was determined using broth microdilution. In a validated murine thigh infection model, HSRs of cefiderocol 2 g q8h as a 3 h IV infusion, ceftazidime/avibactam 2.5 g q8h as a 2 h IV infusion, meropenem 2 g q8h as a 3 h IV infusion or ceftazidime/avibactam/meropenem were administered. Change in bacterial burden relative to baseline and the proportion of isolates in each genotypic group meeting 1-log10 kill endpoint were assessed. Modal MICs (mg/L) ranged from 0.125 to 1 for cefiderocol, 4 to >64 for ceftazidime/avibactam and 2 to >64 for meropenem. Cefiderocol produced >1-log10 of kill against all 18 tested isolates. Meropenem was active against all GES-1 isolates whereas activity against GES-5 and miscellaneous GESs was lacking, consistent with the MICs. Ceftazidime/avibactam was active against all GES-1 and GES-5 isolates and retained activity against 62.5% of miscellaneous GESs including isolates with elevated MICs. For isolates where ceftazidime/avibactam failed, the addition of meropenem restored the in vivo efficacy. As monotherapy, cefiderocol was active in vivo against all tested isolates. The activities of meropenem or ceftazidime/avibactam alone were variable; however, a combination of both was active against all isolates. Cefiderocol and ceftazidime/avibactam/meropenem could be valuable therapeutic options for GES-producing P. aeruginosa infections. Clinical confirmatory data are warranted.

Study of thermal aggregation of growth traits in the branching and elongation phases of a group of genotypes in Coarse wheat (Triticum durum Desf.)

This study was conducted during the agricultural season 2021-2022 at the Agricultural Research and Experiment Station (Sayada area) belonging to the College of Agriculture at the University of Kirkuk, using a randomized complete block design (R.C.B.D) and with three replications, each block included twenty experimental units, the number of genotypes of durum wheat is ( DW38, Bejah-6/SLA, Halio, Mikki, Guayacan, Amedakul s, Fadda98, Icarasha 2, Sham 5, Lahnaucan, Sardar, Bagdad, DW10, Axad5, Simito, Crezo, Corvilla , Iraq Oasis, Surat Kul,Um Rabie), Where the results showed that there are significant differences between the genotypes at the probability level of 1% for the number of branches and the stage of elongation, and the existence of significant differences between the genotypes at the level of probability 1% for the studied traits, where the genetic structure of Sardar exceeded the trait (the number of total fragments, the number of effective fragments, the number of days for the number of branches, 50% spikes parcel) with the highest average, as their average reached (583.20 branches, 526.37 branches, 103.00 days, 16.73 branches, 116.66 days) And the highest average number of branches in the stage (Z21, Z23, Z24, Z25, Z26, Z27, Z28 and Z29) where their average was (1.20, 7.10, 8.46, 9.90, 12.43, 14.36, 15.60, 16.73) and the superiority of the two genetic structures Mikki and Crezo for the characteristic of the area of the flag leaf with the highest average, as their average reached (42.35, 42.18) respectively, while the composition um Rabie with the highest average average plant height, with an average of 83.82, and the genetic makeup of Corvella with the highest average chlorophyll content of the flag paper, with a peak of 47.28 The genetic structure Lahnaucan gives the highest average number of branches in the stage (Z22, Z23, Z24 and (Z25), where their average was (4.20, 7.40, 8.46, 9.60) and the genetic structure Fadda98 gave the highest average number of branches in the stage of Z24) and (Z25, where the average of they are (7.83, 9.70) and the genotype Bejah-6/SLA gives the highest average number of branches in the stage (Z29), where it averaged (16.00) and the superiority of the genetic structure Surat Kul in the elongation stage (for the first decade, the second decade, the third decade, the fourth decade and the fifth decade) The highest average in the number of days, which reached (80.00, 86.66, 92.66, 99.33 and 99.33) and the genetic structure Fadda98 in (the fourth and fifth decade), the highest average in the number of days, reaching (99.33), plants needed to enter the branching stage to the accumulated temperature of 379.0 m0, while plants of all genotypes entered the stage of the maximum number of branches requiring thermal collection of 707.0 m0, and thermal collection required to complete the elongation stage 635.5 m0.

1645-P: Diabetes-Associated Variation in TCF7L2 Is Associated with a Longitudinal Decline in Glucose Tolerance Accompanied by Impaired Suppression of Postprandial Glucagon Secretion

Diabetes-associated alleles in TCF7L2 increase the rate of conversion from prediabetes to type 2 diabetes (T2DM). Multiple studies have shown an association of the T allele at rs7903146 with impaired β-cell function and more recently with defective suppression of α-cell secretion by glucose. However, the temporal relationship of these abnormalities is uncertain. To study the longitudinal changes in islet function, we recruited 140 individuals half of whom were homozygous for the diabetes-associated allele (TT) at rs7903146. The remainder were homozygous for the protective allele (CC). Subjects were studied on 2 occasions, 3 years apart using an oral 75g glucose challenge. This was labelled with 13C-glucose, in concert with intravenously infused tracers to enable measurement of endogenous glucose production, meal appearance and glucose disposal. Indices for insulin secretion and action were estimated using the oral minimal model. Fasting, peak and integrated (443 ± 27 vs. 428 ± 50 mmol/l per 6hr, p = 0.54) post-prandial glucose concentrations did not differ over time in people with the CC genotype. In contrast, post-prandial integrated glucose concentrations increased (441 ± 29 vs. 544 ± 37 mmol/l per 6hr, p &amp;lt; 0.01) in people with the TT genotype. This was accompanied by an increase in integrated postprandial glucagon concentrations (526 ± 39 vs. 618 ± 61 pmol/l per 2hr, p = 0.04) that was not observed in the CC genotype (537 ± 50 vs. 525 ± 65 pmol/l per 2hr, p = 0.07). There were no baseline differences in β-cell function between genotype groups. Disposition Index in both TT (950 ± 130 vs. 868 ± 92 10-4 dl/kg/min per μU/ml, baseline vs. follow-up study respectively, p=0.42) and CC (896 ± 177 vs. 1062 ± 289 10-4 dl/kg/min per μU/ml, p=0.65) groups did not change. This data would suggest that α-cell dysfunction associated with the TT genotype (rs7903146) precedes β-cell dysfunction in people prior to the development of T2DM. Disclosure M.Zeini: None. K.Muthusamy: None. A.M.Egan: None. M.C.Laurenti: None. C.Cobelli: None. C.Dalla man: Research Support; Sanofi-Aventis Deutschland GmbH, Becton, Dickinson and Company. A.Vella: Advisory Panel; Rezolute, Inc., Consultant; Crinetics Pharmaceuticals, Inc., Hanmi Pharm. Co., Ltd., Zealand Pharma A/S, Other Relationship; Novo Nordisk. Funding National Institutes of Health (DK116723)