Glucose 6-Phosphate Accumulates via Phosphoglucose Isomerase Inhibition in Heart Muscle.

RATIONALE: Metabolic and structural remodeling is a hallmark of heart failure. This remodeling involves activation of the mTOR (mammalian target of rapamycin) signaling pathway, but little is known on how intermediary metabolites are integrated as metabolic signals. OBJECTIVE: We investigated the metabolic control of cardiac glycolysis and explored the potential of glucose 6-phosphate (G6P) to regulate glycolytic flux and mTOR activation. METHODS AND RESULTS: We developed a kinetic model of cardiomyocyte carbohydrate metabolism, CardioGlyco, to study the metabolic control of myocardial glycolysis and G6P levels. Metabolic control analysis revealed that G6P concentration is dependent on phosphoglucose isomerase (PGI) activity. Next, we integrated ex vivo tracer studies with mathematical simulations to test how changes in glucose supply and glycolytic flux affect mTOR activation. Nutrient deprivation promoted a tight coupling between glucose uptake and oxidation, G6P reduction, and increased protein-protein interaction between hexokinase II and mTOR. We validated the in silico modeling in cultured adult mouse ventricular cardiomyocytes by modulating PGI activity using erythrose 4-phosphate. Inhibition of glycolytic flux at the level of PGI caused G6P accumulation, which correlated with increased mTOR activation. Using click chemistry, we labeled newly synthesized proteins and confirmed that inhibition of PGI increases protein synthesis. CONCLUSIONS: The reduction of PGI activity directly affects myocyte growth by regulating mTOR activation.

Genome-wide comparative analysis of four Indian Drosophila species.

Comparative analysis of multiple genomes of closely or distantly related Drosophila species undoubtedly creates excitement among evolutionary biologists in exploring the genomic changes with an ecology and evolutionary perspective. We present herewith the de novo assembled whole genome sequences of four Drosophila species, D. bipectinata, D. takahashii, D. biarmipes and D. nasuta of Indian origin using Next Generation Sequencing technology on an Illumina platform along with their detailed assembly statistics. The comparative genomics analysis, e.g. gene predictions and annotations, functional and orthogroup analysis of coding sequences and genome wide SNP distribution were performed. The whole genome of Zaprionus indianus of Indian origin published earlier by us and the genome sequences of previously sequenced 12 Drosophila species available in the NCBI database were included in the analysis. The present work is a part of our ongoing genomics project of Indian Drosophila species.

Photo-reorganization of 3-alkoxy-6-chloro-2-(benzo[b]thiophen-2-yl)-4H-chromen-4-ones: a green and convenient synthesis of angular pentacyclics.

Photo-reorganization of 3-alkoxy-6-chloro-2-(benzo[b]thiophen-2-yl)-4H-chromen-4-ones in methanol with Pyrex filtered UV-light from a medium pressure 125 W Hg-vapor lamp led to the formation of angular pentacyclic compounds (dihydro and aromatic products) along with some rearranged chromenones where the product(s) distribution depended upon the structure of 3-alkoxy groups (methoxy, ethoxy, allyloxy and benzyloxy). The phenyl moiety in the 3-benzyloxy group had a profound effect on the dihydro product(s) formation as the latter was in high yield when the alkoxy group was benzyloxy followed by allyloxy, ethoxy and methoxy groups. The present photochemical study represents a general method for the synthesis of some angular pentacyclic - benzothiophene fused xanthenone derivatives in a single step without using any specific and toxic reagent. The structures of the new organic scaffolds obtained were established by their spectral data (UV, IR and NMR).

Is Drosophila-microbe association species-specific or region specific? A study undertaken involving six Indian Drosophila species.

The present work aims to identify the microbial diversity associated with six Indian Drosophila species using next generation sequencing (NGS) technology and to discover the nature of their distribution across species and eco-geographic regions. Whole fly gDNA of six Drosophila species were used to generate sequences in an Illumina platform using NGS technology. De novo based assembled raw reads were blasted against the NR database of NCBI using BLASTn for identification of their bacterial loads. We have tried to include Drosophila species from different taxonomical groups and subgroups and from three different eco-climatic regions India; four species belong to Central India, while the rest two, D. melanogaster and D. ananassae, belong to West and South India to determine both their species-wise and region-wide distribution. We detected the presence of 33 bacterial genera across all six study species, predominated by the class Proteobacteria. Amongst all, D. melanogaster was found to be the most diverse by carrying around 85% of the bacterial diversity. Our findings infer both species-specific and environment-specific nature of the bacterial species inhabiting the Drosophila host. Though the present results are consistent with most of the earlier studies, they also remain incoherent with some. The present study outcome on the host-bacteria association and their species specific adaptation may provide some insight to understand the host-microbial interactions and the phenotypic implications of microbes on the host physiology. The knowledge gained may be importantly applied into the recent insect and pest population control strategy going to implement through gut microflora in India and abroad.

Phototransformation of 3-alkoxychromenones: regioselective photocyclisation and dealkoxylation.

The phototransformation of some 2-(3-methoxyphenyl)-4H-chromen-4-ones bearing a propynyloxy moiety at the 3-position has been described. On photolysis with pyrex-filtered UV light from a Hg lamp (125 W), these chromenones produced a major amount of 5-ethynyl-2-methoxy-6-oxa-benzo[5,6-c]xanthen-7-ones consisting of an exotic tetracyclic scaffold. These photoproducts have been envisioned to be produced through regioselective ring closure at the 6'-position of the 2-(3'-methoxy)phenyl moiety of the initially formed 1,4-biradical via a γ-H abstraction mechanism. No product whatsoever was observed through ring closure at the 2'-position. This behaviour has been found to be in accordance with the directive influence observed in free radical aromatic substitutions. This regioselective photocyclisation is further supported by calculations made from 3D structures (MM2 program). In addition, during the irradiation of these substrates, 2-(3-methoxyphenyl)-4H-chromen-4-ones were also realised through dealkoxylation. The structures of the substrates and photoproduct(s) have been determined by their spectroscopic (IR, NMR, mass spectrometry) studies.

Absorption and Fluorescent Studies of 3-Hydroxychromones.

The synthesis and spectral studies of variously substituted 3-hydroxychromones have been carried out. A key relationship between the structural motif of synthesized 3-hydroxychromones (3-HCs) and their fluorescent properties was found. The chromones substituted with electron-donating group at 4'-position expressed the red shift of the N(*) and T(*) band and also exhibited the increased fluorescent intensity ratio while the chromones with electron-withdrawing group showed the blue shift of the N(*) and T(*) band. Therefore, these 3-HCs may behave as the possible fluorescent probes.

ATP-dependent citrate lyase Drives Left Ventricular Dysfunction by Metabolic Remodeling of the Heart.

Background: Metabolic remodeling is a hallmark of the failing heart. Oncometabolic stress during cancer increases the activity and abundance of the ATP-dependent citrate lyase (ACL, Acly ), which promotes histone acetylation and cardiac adaptation. ACL is critical for the de novo synthesis of lipids, but how these metabolic alterations contribute to cardiac structural and functional changes remains unclear. Methods: We utilized human heart tissue samples from healthy donor hearts and patients with hypertrophic cardiomyopathy. Further, we used CRISPR/Cas9 gene editing to inactivate Acly in cardiomyocytes of MyH6-Cas9 mice. In vivo, positron emission tomography and ex vivo stable isotope tracer labeling were used to quantify metabolic flux changes in response to the loss of ACL. We conducted a multi-omics analysis using RNA-sequencing and mass spectrometry-based metabolomics and proteomics. Experimental data were integrated into computational modeling using the metabolic network CardioNet to identify significantly dysregulated metabolic processes at a systems level. Results: Here, we show that in mice, ACL drives metabolic adaptation in the heart to sustain contractile function, histone acetylation, and lipid modulation. Notably, we show that loss of ACL increases glucose oxidation while maintaining fatty acid oxidation. Ex vivo isotope tracing experiments revealed a reduced efflux of glucose-derived citrate from the mitochondria into the cytosol, confirming that citrate is required for reductive metabolism in the heart. We demonstrate that YAP inactivation facilitates ACL deficiency. Computational flux analysis and integrative multi-omics analysis indicate that loss of ACL induces alternative isocitrate dehydrogenase 1 flux to compensate. Conclusions: This study mechanistically delineates how cardiac metabolism compensates for suppressed citrate metabolism in response to ACL loss and uncovers metabolic vulnerabilities in the heart.

In vitro and in vivo anti-inflammatory activity of Cupressus torulosa D.DON needles extract and its chemical characterization.

ETHNOPHARMACOLOGICAL RELEVANCE: Cupressus torulosa (family Cupressaceae), widely distributed in the north western Himalayan region of India, is a coniferous aromatic tree with various traditional uses of its aerial parts. Its needles have been used for anti-inflammatory, anticonvulsant, antimicrobial, and wound-healing properties. AIM OF THE STUDY: The study aimed at investigating the previously unknown anti-inflammatory activity of the hydromethanolic extract of the needles employing in vitro and in vivo assays and scientifically validate traditional claim of their use in treatment of inflammation. Chemical characterization of the extract with the aid of UPLCQTOFMS was also of interest. MATERIALS AND METHODS: C. torulosa needles were first defatted with hexane and sequentially extracted with chloroform and 25% aqueous methanol (AM). Since the presence of phenolics (TPCs, 208.21 ± 0.95 mg GAE/g needles) and flavonoids (TFCs, 84.61 ± 1.21 mg QE/g needles) was observed in the AM extract only, it was chosen for biological and chemical examinations. Acute toxicity of the AM extract on female mice was evaluated following the OECD guideline 423. In vitro anti-inflammatory activity of the AM extract was examined using egg albumin denaturation assay while carrageenan-induced paw edema and formalin-induced paw edema models at doses of 100, 200 and 400 mg/kg po were used to determine the in vivo activity of the AM extract on Wistar rats of either sex. The components of the AM extract were analyzed by UPLC-QTOF-MS method using non-targeted metabolomics approach. RESULTS: AM extract was found to be non-toxic at 2000 mg/kg b.w. with no signs of abnormal locomotion, seizures and writhing. The extract demonstrated promising in vitro anti-inflammatory activity (IC50 160.01 µg/mL) compared to standard diclofenac sodium (IC50 73.94 µg/mL) in egg albumin denaturation assay. In carrageenan-induced paw edema and formalin-induced paw edema tests the extract showed significant anti- inflammatory activity (57.28% and 51.04% inhibition of paw edema, respectively) at the dose of 400 mg/kg p.o. after 4 h in comparison to the standard diclofenac sodium which displayed 61.39% and 52.90% inhibition, respectively, at the dose of 10 mg/kg p.o. after 4 h in these models. A total of 63 chemical constituents, majority of them being phenolics, were found in the AM extract of the needles. Two compounds namely monotropein (iridoid glycoside), (±)12-HETE (eicosanoid) and fraxin (coumarin glycoside) were reported to have anti-inflammatory effect. CONCLUSIONS: For the first time our study demonstrated that hydro-methanolic extract of C. torulosa needles exhibit anti-inflammatory activity thereby supporting their traditional use in the treatment of inflammatory disorders. UPLCQTOFMS assisted chemical profile of the extract was also unveiled.

Recreational Marijuana Use and Coronary Artery Dissection: A Case Series.

The cardiovascular effects of marijuana have been shown to be a result of the activation of the CB1 and CB2 receptors located in the myocardium and coronary vasculatures. Adverse cardiovascular consequences of recreational use of marijuana and synthetic cannabinoids include stroke, artery dissection, vasospasm, vasculitis, coronary artery thrombosis, myocarditis/pericarditis, postural hypotension, arrhythmias, and acute heart failure. With marijuana being legalized for medicinal purposes and recreational use in more and more states in the United States, physicians should have a low threshold for the possibility of marijuana being the underlying cause of adverse cardiovascular events. Marijuana has been established to increase sympathetic tone and cause blood pressure elevations and severe coronary artery spasm (CAS). Some studies have even indicated that the risks of heart attack, atrial fibrillation, and ischemic stroke are several times higher within an hour of marijuana use. This case series discusses three female patients, aged 27, 39, and 53 years, who were known to smoke marijuana consistently. These patients all presented with signs and symptoms of acute coronary syndrome (ACS) less than 12 hours after smoking recreational marijuana. All patients endorsed smoking marijuana a few hours prior to the onset of chest pain and less than 12 hours prior to the presentation, which was confirmed by a positive urine drug screen (UDS). Coronary artery angiograms revealed coronary artery dissection in the proximal left circumflex (LCX) artery, the mid-distal left anterior descending (LAD) artery, and mid-LAD in the 27 y/o, 39 y/o, and 53 y/o patients respectively. The average age of spontaneous coronary artery dissection (SCAD) cases ranges between 35-40 years. Women account for more than 70% of cases; it is thought to be due to higher levels of estrogen in women, which alters the normal arterial wall architecture. Additionally, the excessive use of marijuana resulting in CAS further increases the susceptibility to spontaneous dissection in female patients.

Atypical Intracavitary Cardiac Mass: Tumor or Thrombus?

Cardiac masses are a rare finding, with most masses found being thrombi or vegetations. Still, some masses are suspected to be a tumor based on multiple characteristics such as size, location, mobility, and the tumor's hemodynamic effects. Cardiovascular magnetic resonance (CMR) and CT have been shown to differentiate a tumor from a thrombus based on tissue characteristics of the mass. Here we highlight the role of contrast perfusion imaging on echocardiography in identifying the malignant potential of a cardiac mass. This case report demonstrates the effectiveness of contrast imaging with a transesophageal echocardiogram in identifying the etiology of cardiac masses without the need of CMR or cardiac CT, which helps save between $100-1207 of hospital costs. Besides the cost-benefit, the use of non-invasive and easily accessible imaging like echocardiogram enables smaller facilities with limited resources to diagnose and hence further manage patients with cardiac masses.

Hydroxyenone Derivatives: In vitro Anti-malarial and Docking Studies against P. falciparum.

METHODS: A series of 1-{2-(prop-2-ynyloxy)aryl}-3-hydroxy-3-(4'-trifluoromethylphenyl) prop-2-en-1-ones obtained by photo-irradiation of 2-{2-(prop-2-ynyloxy)benzoyl}-3-(4- trifluorome-thyl-phenyl)oxiranes (that were characterized by spectral studies: FT-IR, 1H NMR, 13C NMR and Mass analysis) was screened for the anti-malarial activity by evaluating against chloroquine-sensitive P. falciparum (CD7). The molecular docking studies using AutoDock Vina were also performed to further ascertain the efficacy of these compounds with PDB:4ORM. RESULTS: Among these, the hydroxyenone derivatives 2b, 2c and 2a exhibited very potent antimalarial activity that was clearly evinced by the results of molecular docking. Binding energies of hydroxyenone compounds were calculated and found in the range of -10.4 to -9.0 kcal/mol. CONCLUSION: Compound 2b had the strongest binding affinity with docking score of -10.4 kcal/mol.

Whole genome sequence resource of Indian Zaprionus indianus.

This article documents the whole genome sequence information of the Indian Zaprionus indianus, a member of the fruit fly family Drosophilidae. The sequences were generated on an Illumina platform and reads and whole genome sequence submitted to NCBI to the SRA and BioProject databases, respectively. This is the first Indian Z. indianus whole genome (draft) submitted to the sequence repository with SRA reads. The details of methodology, assembly statistics and functional annotation are presented in this work.

Development of Computer Algorithm for Editing of Next Generation Sequencing Metagenome Data.

The successful implementation of the advanced sequencing technology, the next generation sequencing (NGS) motivates scientists from diverse fields of biological research especially from genomics and transcriptomics in generating large genomic data set to make their analysis more robust and come up with strong inference. However, exploiting this huge genomic data set becomes a challenge for the molecular biologists. To corroborate this problem, computational software and hardware are being developed in parallel and become an integral part of life science. While executing the "Genomics project of Indian Drosophila species," we found strings of Ns in the whole genome sequences generated on Illumina platform. The present article aims at developing a computer algorithm (MATLAB and Python based) for editing raw sequences mainly eliminating bad residues before submitting to the publicly accessible sequence repository. These algorithms will be helpful to life scientists for analyzing large amount of biological data in short span of time.