GmMYB176 Interactome and Regulation of Isoflavonoid Biosynthesis in Soybean

MYB transcription factors are one of the largest transcription factor families characterized in plants. They are classified into four types: R1 MYB, R2R3 MYB, R3 MYB and R4 MYB. GmMYB176 is an R1MYB transcription factor that regulates Chalcone synthase (CHS8) gene expression and isoflavonoid biosynthesis in soybean. Silencing of GmMYB176 suppressed the expression of the GmCHS8 gene and reduced the accumulation of isoflavonoids in soybean hairy roots. However, overexpression of GmMYB176 does not alter either GmCHS8 gene expression or isoflavonoid levels suggesting that GmMYB176 alone is not sufficient for GmCHS8 gene regulation. I hypothesized that GmMYB176 acts cooperatively with another factor(s) for the regulation of GmCHS8 gene expression and it may also regulate other isoflavonoid biosynthetic genes in soybean. The objective of this research was to identify the GmMYB176 interactome for GmCHS8 gene regulation and elucidate the role of GmMYB176 in isoflavonoid biosynthesis in soybean. GmMYB176 interacting proteins were identified using two translational fusion baits (GmMYB176-YFP and YFP-GmMYB176) by co-immunoprecipitation, followed by liquid chromatography-tandem mass spectrometry. The interaction of selected candidates with GmMYB176 was validated in planta and their DNA binding activities determined. GmMYB176 may form a transcriptional complex with Gm04bZIP and/or Gm05bZIP for the regulation of GmCHS8 gene expression. RNA-seq and metabolomics analyses of soybean hairy roots in which GmMYB176 was either silenced or over-expressed revealed that GmMYB176 regulates multiple genes in the isoflavonoid biosynthesis pathway, affecting the production of metabolites in phenylpropanoid pathway such as phenylalanine, liquiritigenin, daidzin, genistin, glycitein, and glyceollin. The knowledge and information generated on the role of GmMYB176 in the regulation of isoflavonoid biosynthesis will allow genetic manipulation of isoflavonoid level in soybean and/or introduce isoflavonoid pathway in non-legumes

Three dimensional Aero-Structural Shape Optimization of Turbomachinery Blades

Aero-structural optimization of gas turbine blades is a very challenging task, given e.g. three dimensional nature of the flow, stringent performance requirements, structural and manufacturing considerations, etc. The current research work addresses this challenge by development and implementation of structural shape optimization module and integrating it with an aerodynamic shape optimization module to form an automated aero-structural optimization procedure. The optimizer combines a Multi-Objective Genetic Algorithm (MOGA), with a Response Surface Approximation (RSA) of the Artificial Neural Network (ANN) type. During the optimization process, each objective function and constraint is approximated by an individual ANN, which is trained and tested using an aerodynamic as well as a structure database composed of a few high fidelity flow simulations (CFD) and structure analysis (CSD) that are obtained using ANSYS Workbench 11.0. Addition of this multiple ANN technique to the optimizer greatly improves the accuracy of the RSA, provides control over handling different design variables and disciplines. The described methodology is then applied to the aero-structural optimization of the E/TU-3 turbine blade row and stage at design conditions to improve the aerodynamic and structural performance of the turbomachinery blades by optimizing the stacking curve. The proposed methodology proved quite successful, flexible and practical with significant increase in stage efficiency and decrease in equivalent stress

Plasma hs-crp level as a predictor of left ventricular function & myocardial reperfusion injury in patients with acute myocardial infarction-thrombolysed

1. In the estimation of High sensitive- C –reactive protein, the high risk group predominated at 52% with a mean of 6.72±1.76, intermediate risk - 25% with a mean of 2.02±0.083 and low risk - 23% with a mean of 0.75±0.239. 2. In all the groups male patients predominated. 3. In the low and intermediate group the window period was predominantly in the range of 3- 6 hrs, in high risk group it was > 6 hrs. 4. The mean ejection fraction in the low risk was 57±11, intermediate risk was 54.12±10.76 and in high risk group it was 50±10.8. 5. The reperfusion injury was only 4% in the low risk group, 8% in the intermediate risk and it was higher in the high risk group at 42%. 6. The most common site of acute myocardial infarction was extensive anterior wall myocardial infarction followed by inferior wall myocardial infarction. 7. Among women, 55.55% of post menopausal women had acute myocardial infarction. 8) Oral contraceptive pill was a risk factor in 1 patient (5.5%). 9. Smoking was one of the major risk factor in 79.2% of male patients. 10. Obesity was found to be a risk factor in 22% of patients.55.55% among female were obese. 11. Diabetes was found in 29% of patients of which 79.3% were male. 12. Hypertension was seen in 49% patients. Of these 79.59% were male. CONCLUSION : In this Present study it is found that Ejection Fraction is lower in the high CRP group. Reperfusion injury is lower in the low and intermediate risk group according to the High Sensitive C –Reactive Protein assay. As echocardiogram was done only on day three after Acute myocardial infarction, a lot of stunned myocardium may be present, which is the limitation of this study

Comparative study of fatty meal Versus drotaverine hydrochloride Versus hyoscine-n-butylbromide for duodenal antimotility and ease of cannulation during Endoscopic Retrograde Cholangiopancreatography (ERCP).

INTRODUCTION : Endoscopic retrograde cholangiopancreatography (ERCP) is in clinical practice since 1968 for diagnosis and treatment of pancreatobiliary diseases. ERCP is a technically demanding procedure that requires considerable amount of training to be performed safely. Common bile duct (CBD) and/or the pancreatic duct (PD) cannulation is the critical step. Failed intervention or an aborted procedure is due to failure of cannulation of the desired ducts. A side viewing duodenoscope of 120cms is necessary for cannulation of normal gastrointestinal anatomy, whereas a forward viewing duodenoscope may be necessary in case of altered anatomy like Billroth II anastomoses, Roux‐en Y anastomosis, Choledochojejunostomy and Hepaticojejunostomy. In surgically altered anatomy, usage of a double or “short” double‐balloon and single balloon enteroscopy has achieved a higher successful cannulation rates. For diagnostic and therapeutic interventions like sphincterotomy, stent placement and stone extraction, cannulation of the ampulla of Vater and wire access of the ampulla is the prime requirement. A higher rate of successful cannulation without complications usually depends on the expertise of the endoscopist. Those with lesser experience have a much lower rate of success and higher occurrence of complications. The anatomy typically predisposes to pancreatic duct cannulation as the pancreatic duct enters the ampulla in a straight fashion. Edema or strictures in the small bowel, surgically altered anatomy, blood or excessive fluid in the lumen and periampullary diverticulum increases the difficulty of the procedure. Prolonged cannulation often results in increased patient morbidity, unnecessary radiation exposure and anxiety for endoscopists. With normal anatomy, various technical methods have been adopted to facilitate cannulation at ERCP based on technical and pharmacological aspects. AIM : To compare the effect of Fatty meal versus Drotaverine hydrochloride versus Hyoscine‐N‐butyl bromide: On duodenal contraction rate, To identify the ampulla, Time for cannulation, Adverse events. CONCLUSION : In the current study fatty meal is not inferior to the conventionally used Hyoscine‐N‐butyl bromide or its alternate Drotaverine for its anti‐motility effect on the duodenum during ERCP. The cannulation time is no different within the groups. The identification of ampulla was easier with the fatty meal group. There is a possibility of higher increase in pulse rate and blood pressure in patients with ischemic heart disease with the use of Hyoscine‐N‐butyl bromide. Further larger randomized trials are required to confirm the efficacy of fatty meal in ERCP

GmMYB176 Regulates Multiple Steps in Isoflavonoid Biosynthesis in Soybean

Isoflavonoids are a group of plant natural compounds synthesized almost exclusively by legumes, and are abundant in soybean seeds and roots. They play important roles in plant-microbial interactions and the induction of nod gene expression in Rhizobia that form nitrogen-fixing nodules on soybean roots. Isoflavonoids also contribute to the positive health effects associated with soybean consumption by humans and animals. An R1 MYB transcription factor GmMYB176 regulates isoflavonoid biosynthesis by activating chalcone synthase (CHS) 8 gene expression in soybean. Using a combination of transcriptomic and metabolomic analyses of GmMYB176-RNAi silenced (GmMYB176-Si), GmMYB176-overexpressed (GmMYB176-OE), and control soybean hairy roots, we identified a total of 33 differentially expressed genes (DEGs) and 995 differentially produced metabolite features (DPMF) in GmMYB176-Si hairy roots, and 5727 DEGs and 149 DPMFs in GmMYB176-OE hairy roots. By a targeted approach, 25 isoflavonoid biosynthetic genes and 6 metabolites were identified as differentially regulated in GmMYB176-OE and GmMYB176-Si soybean hairy roots. Taken together, our results demonstrate the complexity of isoflavonoid biosynthesis in soybean roots and suggest that a coordinated expression of pathway genes, substrate flux and product threshold level may contribute to the dynamic of the pathway regulation

Splicing is an alternate oncogenic pathway activation mechanism in glioma

High-grade diffuse glioma (HGG) is the leading cause of brain tumour death. While the genetic drivers of HGG have been well described, targeting these has thus far had little impact on survival suggesting other mechanisms are at play. Here we interrogate the alternative splicing landscape of pediatric and adult HGG through multi-omic analyses, uncovering an increased splicing burden compared with normal brain. The rate of recurrent alternative splicing in cancer drivers exceeds their mutation rate, a pattern that is recapitulated in pan-cancer analyses, and is associated with worse prognosis in HGG. We investigate potential oncogenicity by interrogating cancer pathways affected by alternative splicing in HGG; spliced cancer drivers include members of the RAS/MAPK pathway. RAS suppressor neurofibromin 1 is differentially spliced to a less active isoform in >80% of HGG downstream from REST upregulation, activating the RAS/MAPK pathway and reducing glioblastoma patient survival. Overall, our results identify non-mutagenic mechanisms by which cancers activate oncogenic pathways which need to accounted for in personalized medicine approaches

Genome-wide identification and localization of chalcone synthase family in soybean (Glycine max [L]Merr)

Abstract Background Soybean is a paleopolyploid that has undergone two whole genome duplication events. Gene duplication is a type of genomic change that can lead to novel functions of pre-existing genes. Chalcone synthase (CHS) is the plant-specific type III polyketide synthase that catalyzes the first committed step in (iso)flavonoid biosynthesis in plants. Results Here we performed a genome-wide search of CHS genes in soybean, and identified 21 GmCHS loci containing 14 unique GmCHS (GmCHS1-GmCHS14) that included 5 newly identified GmCHSs (GmCHS10-GmCHS14). Furthermore, 3 copies of GmCHS3 and 2 copies of GmCHS4 were found in soybean. Analysis of gene structure of GmCHSs revealed the presence of a single intron in protein-coding regions except for GmCHS12 that contained 3 introns. Even though GmCHS genes are located on 8 different chromosomes, a large number of these genes are present on chromosome 8 where they form 3 distinct clusters. Expression analysis of GmCHS genes revealed tissue-specific expression pattern, and that some GmCHS isoforms localize in the cytoplasm and the nucleus while other isoforms are restricted to cytoplasm only. Conclusion Overall, we have identified 21 GmCHS loci with 14 unique GmCHS genes in the soybean genome. Their gene structures and genomic organization together with the spatio-temporal expression and protein localization suggest their importance in the production of downstream metabolites such as (iso)flavonoids and their derived phytoalexins

Development & standardization of an in-house IgM indirect ELISA for the detection of parvovirus B19 infections

Background & objectives: Parvovirus B19 infections occur worldwide; the infection is acquired early in childhood but could occur later. B19 is reported to cause infection in childhood febrile illnesses, and arthropathies in adults and children and in end-stage renal disease (ESRD) seen in adults. This study was designed to develop an in-house IgM indirect ELISA for serological screening among patients and controls, and to compare ELISA results with those of nested polymerase chain reaction (nPCR) assay. Methods: An in-house IgM indirect ELISA was standardized using peptide sequence of VP1/VP2 region of parvovirus B19. A total of 201 children and adult with febrile illnesses, 216 individuals with non-traumatic arthropathies, 201 cases of chronic anaemia associated with ESRD and 100 healthy controls were tested. Serum was separated from the blood and subsequently used for DNA extraction. The nested polymerase chain reaction (nPCR) for the detection of B19V DNA was performed using primers targeting the overlapping region of VP1/VP2 capsid protein genes. Results: A total of 618 samples were tested for parvovirus B19 by an in-house IgM indirect ELISA. Among these samples, six were positive by in-house ELISA. The inter-rater agreement between ELISA and PCR assays was calculated using kappa coefficient analysis. The value of κ was 0.77 and the strength of agreement was 'good' (P<0.001). Interpretation & conclusions: The in-house IgM indirect ELISA was found to be simple with high sensitivity and specificity when compared with nPCR and could be used as an alternative to expensive commercial kits in resource-poor settings

Novel Insights on Hantavirus Evolution: The Dichotomy in Evolutionary Pressures Acting on Different Hantavirus Segments

<div><p>Background</p><p>Hantaviruses are important emerging zoonotic pathogens. The current understanding of hantavirus evolution is complicated by the lack of consensus on co-divergence of hantaviruses with their animal hosts. In addition, hantaviruses have long-term associations with their reservoir hosts. Analyzing the relative abundance of dinucleotides may shed new light on hantavirus evolution. We studied the relative abundance of dinucleotides and the evolutionary pressures shaping different hantavirus segments.</p><p>Methods</p><p>A total of 118 sequences were analyzed; this includes 51 sequences of the S segment, 43 sequences of the M segment and 23 sequences of the L segment. The relative abundance of dinucleotides, effective codon number (ENC), codon usage biases were analyzed. Standard methods were used to investigate the relative roles of mutational pressure and translational selection on the three hantavirus segments.</p><p>Results</p><p>All three segments of hantaviruses are CpG depleted. Mutational pressure is the predominant evolutionary force leading to CpG depletion among hantaviruses. Interestingly, the S segment of hantaviruses is GpU depleted and in contrast to CpG depletion, the depletion of GpU dinucleotides from the S segment is driven by translational selection. Our findings also suggest that mutational pressure is the primary evolutionary pressure acting on the S and the M segments of hantaviruses. While translational selection plays a key role in shaping the evolution of the L segment. Our findings highlight how different evolutionary pressures may contribute disproportionally to the evolution of the three hantavirus segments. These findings provide new insights on the current understanding of hantavirus evolution.</p><p>Conclusions</p><p>There is a dichotomy among evolutionary pressures shaping a) the relative abundance of different dinucleotides in hantavirus genomes b) the evolution of the three hantavirus segments.</p></div