Design, synthesis, molecular docking, and biological evaluation of coumarin-thymidine analogs as potent anti-TB agents.

With the intent to discover new antituberculosis (TB) compounds, coumarin-thymidine analogs were synthesized using second-order nucleophilic substitution reactions of bromomethyl coumarin with thymidine. The newly synthesized coumarin-thymidine conjugates (1a-l) were characterized using IR, NMR, GC-MS, and CHN elemental analysis. The novel conjugates were found to exhibit potent anti-TB activity against the Mycobacterium tuberculosis H37 Rv strain, with minimum inhibitory concentrations (MIC) of the active compounds ranging between 0.012 and 0.482 µM. Compound 1k was established as the most active candidate with a MIC of 0.012 µM. The toxicity study on HEK cells confirmed the nontoxic nature of compounds 1e, 1h, 1i, 1j, and 1k. Also, the most active compounds (1k, 1j, and 1e) were stable in the pH range from 2.5 to 10, indicating compatibility with the biophysical environment. Based on the pKa studies, compounds 1k, 1j, and 1e are capable of crossing lipid-membrane barriers and acting on target cells. Molecular docking studies on the M. tuberculosis ß-oxidation trifunctional enzyme (PDB ID: 7O4V) were conducted to investigate the mechanisms of anti-TB activity. All compounds showed excellent hydrogen binding interactions and exceptional docking scores against M. tuberculosis, which was in accordance with the results. Compounds 1a-l possessed excellent affinity to proteins, with binding energies ranging from -7.4 to -8.7 kcal/mol.

ON donor tethered copper (II) and vanadium (V) complexes as efficacious anti-TB and anti-fungal agents with spectroscopic approached HSA interactions.

Antimicrobial drug resistance poses a significant threat worldwide, hence triggering an urgent situation for developing feasible drugs. 3D-transition metal coordination complexes being multifaceted, offer tremendous potency as drug candidates. However, there are fewer reports on non-toxic and safe transition metal complexes; therefore, we hereby attempted to develop novel copper and vanadium-based therapeutic agents. We have synthesised six metal complexes viz., [VVO2(Quibal-INH)] (1), [CuII(Quibal-INH)2] (2), [VVO(Quibal-INH) (cat)] (3), [CuII(Quibal-INH) (cat)] (4), [VVO(Quibal-INH) (bha)] (5) and [CuII(Quibal-INH) (bha)] (6). Quibal-INH (L) is an ON bidentate donor ligand synthesized from Schiff base reaction between 4-(2-(7-chloroquinolin-3-yl)vinyl)benzaldehyde (Quibal) and Isoniazid (INH). The synthesized compounds were characterized using analytical techniques involving ATR-IR, UV-Vis, EPR, 1H NMR, 13C NMR, and 51V NMR. Ligand (L) and compound 3 exhibited moderate growth inhibitory activity towards Candida albicans and Cryptococcus neoformans fungal species. Compound 6 has been identified as active against the above fungal species with no toxicity and hemolysis activity on the healthy cells. Compound 5 exhibited significant activity against the Mycobacterium tuberculosis H 37 R v strain. Further, compounds 4, 5 and 6 exhibited excellent free radical scavenging activity. All the developed compounds were found to exhibit stability over a wide range of pH conditions. The complexes were additionally studied for their interaction with human serum albumin (HSA) with the UV-vis spectroscopic technique.

Snake Venom-specific Phospholipase A2: A Diagnostic Marker for the Management of Snakebite Cases.

Background: Snakebites are a common cause of morbidity and mortality, especially in tropical countries. Snakebites in any community are managed based on the clinical features and intravenous administration of antisnake venom (ASV). The administration of ASV is either deficient or given in excess based on clinical decisions and whole blood clotting test results. The present study is designed to analyze the level of snake venom component in the blood of snakebite in association with the clinical features. Patients and methods: Blood samples were collected from the patients admitted to Karnataka Institute of Medical (KIMS) hospital with a history of snakebite considering the inclusion criteria. Serum was collected from the blood of snakebite patients before and after ASV and used to assess the level of venom-specific phospholipase A2 (PLA2) enzyme using the enzyme-linked immunosorbent assay (ELISA) method. Results: Quantitative ELISA results revealed that the snake venom-specific PLA2 in the victim's blood was in the range of 0.3-1.27 mg/mL before the administration of ASV. However, the concentration of PLA2 after 24 hours of ASV administration was decreased in most of the patients. Further, it was observed that envenomation complications were directly proportional to the amount of snake venom-specific PLA2 found in the blood of the snakebite patient. Conclusion: The study concludes that snake venom-specific PLA2 in the blood of snakebite patients could be used as a reliable venom marker, which helps in determination of appropriate ASV dosage in snakebite patients. How to cite this article: Kaulgud RS, Hasan T, Vanti GL, Veeresh S, Uppar AP, Kurjogi MM. Snake Venom-specific Phospholipase A2: A Diagnostic Marker for the Management of Snakebite Cases. Indian J Crit Care Med 2022;26(12):1259-1266.

Prevalence of hypertension and its associated risk factors in Dharwad population: A cross-sectional study.

The study aims to determine the prevalence of hypertension and associated risk factors. The study includes 9754 participants, out of which 6403 were found to be associated with hypertension. Among 6403 participants 27.75% were newly diagnosed with hypertension during examination. The present study showed, age as one of the significant risk factors for prevalence of hypertension. Further observations revealed that, the prevalence of hypertension was higher in alcohol-intake, tobacco-smoking/chewing participants and sedentary life style is also one of significant risk factor for hypertension. Overall increased rate of hypertension pose a biggest challenge for health sector in Dharwad district.

Genome-wide Characterization and Expression Analysis of Sugar Transporter Family Genes in Woodland Strawberry.

In higher plants, sugars are nutrients and important signal molecules. Sugar transporters (STs) facilitate sugar transport across membranes and are associated with loading and unloading of the conducting complex. Strawberry ( Duchesne ex Rozier) is one of the most economically important and widely cultivated fruit crop and a model plant among fleshy fruits worldwide. In this study, 66 woodland strawberry ( L.) ST (FvST) genes were identified and further classified into eight distinct subfamilies in the woodland strawberry genome based on the phylogenetic analysis. In the promoter sequences of FvST gene families, a search for -regulatory elements suggested that some of them might probably be regulated by plant hormones (e.g., salicylic acid, abscisic acid, and auxin), abiotic (e.g., drought, excessive cold, and light), and biotic stress factors. Exon-intron analysis showed that each subfamily manifested closely associated gene architectural features based on similar number or length of exons. Moreover, to comprehend the potential evolution mechanism of FvST gene family, the analysis of genome duplication events was performed. The segmental and tandem duplication analysis elucidated that some of ST genes arose through whole-genome duplication (WGD) or segmental duplication, accompanied by tandem duplications. The expression analysis of 24 FvST genes in vegetative and during fruit development has shown that the expression of several ST genes was tissue and developmental stage specific. Generally, our findings are important in understanding of the allocation of photo assimilates from source to sink cell and provide insights into the genomic organization and expression profiling of FvST gene families in woodland strawberry.

Synthesis and evaluation of novel coumarin-oxime ethers as potential anti-tubercular agents: Their DNA cleavage ability and BSA interaction study.

As a contribution to the development of novel coumarin-oxime ether conjugates with therapeutically interesting properties, a series of coumarin-oxime ether (1a-1j) was synthesised using SN2 reaction of bromomethyl coumarins with butane-2,3-dione monoxime. Invitro anti-tuberculosis activityagainstMTBH37Rv strain was established for the coumarin-oxime ether (1a-1j). Most of the compounds exhibited significant activity with minimum inhibitory concentration (MIC)in the range of 0.04-3.12 µg mL-1. Compound (1h) was identified as a hit candidate exhibiting MIC of 0.04 µg mL-1, closer to the MIC value of Isoniazid (0.02 µg mL-1), a commercially available drug for the treatment of tuberculosis. Compound 1h also displayed a low level of toxicity in Vero cells along with a good safety profile in vitro. Compounds that showed potent anti-tubercular activity were also found to cleave DNA more efficiently and thereby exhibit nuclease activity. The most active compound (1h) was further studied to deduce the mode of interaction with model serum protein, bovine serum albumin (BSA).

Synthesis of coumarin-theophylline hybrids as a new class of anti-tubercular and anti-microbial agents.

A series of novel coumarin-theophylline hybrids were synthesized and examined for their anti-tubercular activity in vitro against Mycobacterium tuberculosis H37Rv, anti-microbial activity in vitro against gram-positive bacteria (Staphylococcus aureus) and gram-negative bacterias (Escherichia coli, Salmonella typhi) as well as fungi (Candida albicans). The compound (3a) has shown excellent anti-tubercular activity with MIC of 0.12 µg/mL. Electron donating compounds (3a, 3f) have displayed significant anti-microbial activity. The compounds have also been precisely elucidated using single crystal X-ray diffraction techniques. Molecular docking study has been performed against 4DQU enzyme of Mycobacterium tuberculosis showed good binding interactions and is in agreement with the in vitro results.

Design, synthesis, and evaluation of new α-aminonitrile-based benzimidazole biomolecules as potent antimicrobial and antitubercular agents.

The study explores the one-pot synthesis of novel α-aminonitriles by reacting 4-[(1H-benzimidazol-2-yl)methoxy]benzaldehyde, substituted anilines and sodium cyanide using a catalytic amount of copper dipyridine dichloride (CuPy2 Cl2 ) and employing the Strecker reaction under mild conditions. All the synthesized compounds were screened for antimicrobial and antitubercular activity. The promising lead compounds 4d and 4e were identified, with MIC values ranging between 3.9 and 7.8 µg/mL against different bacterial strains. Compounds 4c-e and 4g also showed good antifungal activities against the tested fungal strain. Among those tested, compound 4e exhibited excellent antitubercular activity (MIC 0.05 µg/mL) with a low level of cytotoxicity, suggesting that compound 4e is a promising lead for subsequent investigations in search for new antitubercular agents.

Grapevine immune signaling network in response to drought stress as revealed by transcriptomic analysis.

Drought is a ubiquitous abiotic factor that severely impedes growth and development of horticulture crops. The challenge postured by global climate change is the evolution of drought-tolerant cultivars that could cope with concurrent stress. Hence, in this study, biochemical, physiological and transcriptome analysis were investigated in drought-treated grapevine leaves. The results revealed that photosynthetic activity and reducing sugars were significantly diminished which were positively correlated with low stomatal conductance and CO2 exchange in drought-stressed leaves. Further, the activities of superoxide dismutase, peroxidase, and catalase were significantly actuated in the drought-responsive grapevine leaves. Similarly, the levels of abscisic acid and jasmonic acid were also significantly increased in the drought-stressed leaves. In transcriptome analysis, 12,451 differentially-expressed genes (DEGs) were annotated, out of which 8021 DEGs were up-regulated and 4430 DEGs were down-regulated in response to drought stress. In addition, the genes encoding pathogen-associated molecular pattern (PAMP) triggered immunity (PTI), including calcium signals, protein phosphatase 2C, calcineurin B-like proteins, MAPKs, and phosphorylation (FLS2 and MEKK1) cascades were up-regulated in response to drought stress. Several genes related to plant-pathogen interaction pathway (RPM1, PBS1, RPS5, RIN4, MIN7, PR1, and WRKYs) were also found up-regulated in response to drought stress. Overall the results of present study showed the dynamic interaction of DEG in grapevine physiology which provides the premise for selection of defense-related genes against drought stress for subsequent grapevine breeding programs.

Insights into grapevine defense response against drought as revealed by biochemical, physiological and RNA-Seq analysis.

Grapevine is an important and extensively grown fruit crop, which is severely hampered by drought worldwide. So, comprehending the impact of drought on grapevine genetic resources is necessary. In the present study, RNA-sequencing was executed using cDNA libraries constructed from both drought-stress and control plants. Results generated 12,451 differentially expressed genes (DEGs), out of which 8,021 genes were up-regulated, and 4,430 were down-regulated. Further physiological and biochemical investigations were also performed to validate the biological processes associated with the development of grapevine in response to drought stress. Results also revealed that decline in the rate of stomatal conductance, in turn, decrease the photosynthetic activity and CO2 assimilation in the grapevine leaves. Reactive oxygen species, including stress enzymes and their related proteins, and secondary metabolites were also activated in the present study. Likewise, various hormones also induced in response to drought stress. Overall, the present study concludes that these DEGs play both positive and negative roles in drought tolerance by regulating various biological pathways of grapevine. Nevertheless, our findings have provided valuable gene information for future studies of abiotic stress in grapevine and various other fruit crops.

Epidemiology of Bovine Mastitis in Cows of Dharwad District.

Bovine mastitis is very common in cows of both developed and developing countries. The prevalence of clinical and subclinical mastitis (SCM) varies from region to region. Hence, the present study was carried out to determine the prevalence of mastitis using three diagnostic tests by considering different risk factors like age, lactation, breed, season, quarters, and herd. The results showed that surf field mastitis test (SFMT) is the most sensitive test for diagnosis of bovine mastitis, the older age and cows with later part of lactation period were more prone to bovine mastitis, and exotic breeds like Holstein freshen (HF) were more susceptible to bovine mastitis. The highest incidence of mastitis was recorded in monsoon season. The prevalence of subclinical and clinical mastitis was more in single and two quarters, respectively, and the rate of bovine mastitis was more in unorganized herds. The study concluded that SCM is directly associated with age, lactation period, and environmental factors of the cow and clinical mastitis is more associated with breed of the cow and environmental conditions.

In silico characterization of putative drug targets in Staphylococcus saprophyticus, causing bovine mastitis.

The bovine mastitis caused by coagulase negative staphylococci (CNS) has increased in many herds of urban and rural areas of India. Emergence of multi drug resistant bacteria has further made its management more complex and serious. Therefore, innovation of novel specific drug for the treatment of disease caused by particular organism remained to be a challenge. Hence, in the present study a bacterium was isolated from milk of the cow with bovine mastitis and was identified as S. saprophyticus, 44 pathways of S. saprophyticus retrieved (KEGG) from web server were found to be non homologous to the host Bos taurus, out of which 39 pathways were found to be in cytoplasm, 2 in cell wall and 3 in the cell membrane. The knowledge of the present study could make the drug discovery easier which have high affinity to the target site of the causative organism.