Pyridine-2,6-Dicarboxamide Proligands and their Cu(II)/Zn(II) Complexes Targeting Staphylococcus Aureus for the Attenuation of In Vivo Dental Biofilm.

In the pursuit to combat stubborn bacterial infections, particularly those stemming from gram-positive bacteria, this study is an attempt to craft a precision-driven platform characterized by unparalleled selectivity, specificity, and synergistic antimicrobial mechanisms. Leveraging remarkable potential of metalloantibiotics in antimicrobial applications, herein, this work rationally designs, synthesizes, and characterizes a new library of Pyridine-2,6-dicarboxamide ligands and their corresponding transition metal Cu(II)/Zn(II) complexes. The lead compound L11 demonstrates robust antibacterial properties against Staphylococcus aureus (Minimum Inhibitory Concentration (MIC) = 2-16 µg mL-1), methicillin and vancomycin-resistant S. aureus (MIC = 2-4 µg mL-1) and exhibit superior antibacterial activity when compared to FDA-approved vancomycin, the drug of last resort. Additionally, the compound exhibits notable antimicrobial efficacy against resistant enterococcus strains (MIC = 2-8 µg mL-1). To unravel mechanistic profile, advanced imaging techniques including SEM and AFM are harnessed, collectively suggesting a mechanistic pathway involving cell wall disruption. Live/dead fluorescence studies further confirm efficacy of L11 and its complexes against S. aureus membranes. This translational exploration extends to a rat model, indicating promising in vivo therapeutic potential. Thus, this comprehensive research initiative has capabilities to transcends the confines of this laboratory, heralding a pivotal step toward combatting antibiotic-resistant pathogens and advancing the frontiers of metalloantibiotics-based therapy with a profound clinical implication.

Development of Simplified Recombinase Polymerase Amplification Assay for Rapid and Robust Detection of Citrus Yellow Vein Clearing Virus.

Citrus is an economically important fruit crop, belongs to family Rutaceae, cultivated commercially in over 130 countries, which holds a leading profitable position in the international market. The most important citrus varieties are mandarins, oranges, lemons, sweet limes, grapefruits and pomelos. Citrus yellow vein clearing virus (CYVCV) is an important graft transmissible plant pathogen known to reduce productivity of citrus fruits due to its predominant association and widespread occurrence. Requirement of fast, reliable, efficient & economical CYVCV indexing assay is a prerequisite for production of healthy planting material. Currently, nucleic acid isolation and thermal cycler-based assay available for CYVCV indexing is a cumbersome lab intensive method. The present study was undertaken to develop and validate reverse transcription-recombinase polymerase amplification (RT-RPA) assay requiring no tedious RNA isolation, separate cDNA synthesis and costlier instrument like thermo-cycler. Optimized RT-RPA assay was able to amplify CYVCV up to 10-7 dilution (equivalent to 0.1 pg/µl) with the prepared templates of both RNA and crude saps and showed higher sensitivity in detection of CYVCV infection in field samples as compared to the conventional RT-PCR. Developed RT-RPA assay showed high specificity without any cross-reaction with other citrus pathogens (Indian citrus ringspot virus, citrus yellow mosaic virus, citrus tristeza virus, citrus exocortis viroid and huanglongbing). RT-RPA using crude leaf sap as template is quite simple, robust, highly sensitive, time and cost effective; therefore, it can be used in resource constrained laboratories as screening tool, for field surveys and on-site testing programs in farms, nurseries and biosecurity. Present study, first time reports the development, optimization and validation of crude sap-based RT-RPA assay for the detection of CYVCV infection in citrus plants namely; Kinnow mandarin, Mosambi and Grape fruit.

Fresh and recirculated submarine groundwater discharge zones along the central west coast of India.

The study area receives an average of 2840.0 mm of rainfall within four months every year. A portion of the rainwater is flown to the sea as surface water, and the other part is percolated into the bottom as groundwater. In coastal aquifers, the groundwater is transported to the sea due to a hydraulic gradient, and it contains a significant quantity of dissolved materials and nutrients. SGD processes impact the ocean productivity, mangrove and coral growth, local acidification and many. To isolate the SGD on the central west coast of India, different data was referred. The GWL concerning MSL contributed significantly to demarcating the SGD zones by considering the positive (>0 m) and negative (<0 m) values of GWL concerning above MSL. Thermal images for SST of pre-monsoon and post-monsoon periods of 2020 exhibit cooler surrounded by warmer, which might be the SGD buffering zones in the off-central west coast of India. By considering the results from GWL and SST, 8 SGD beach sites were identified for the further particularized study. The water samples were collected in March 2022, and analyzed using standard procedures and instruments. Fresh and mixing (recirculated) zones have been isolated by piper, hydrochemical facies evolution, and Ca2++Mg2+/K++Na + Vs log Cl- ionic ratio plots. The aquifer water chemical elements are converting possibly due to ionic exchange processes. The decrease in salinity and conductivity observed in the pore water just below the seawater might be due to the influence of freshwater inputs, helping to isolate the fresh SGD and recirculated SGD zones in the study area. Among 8 sites, 3 were found to be fresh SGD sites and 5 were noticed to be mixing/recirculated SGD sites. Most of these Beaches are bounded by hills, which helps to lead the SGD along the central west coast of India.

Development of Polyclonal Antibodies-Based Serological Method for the Detection of Calanthe Mild Mosaic Virus and Application in Virus Certification Programme.

Calanthe mild mosaic virus (CalMMV) infecting orchids is an important potyvirus which is known to cause mild leaf mosaic and flower colour-breaking symptoms in Calanthe and other orchid plants. The present study reports the production of polyclonal antibodies against CalMMV using bacterially expressed recombinant coat protein as immunogen, which in turn would be useful in routine indexing and screening of orchid germplasm. The coat protein (CP) gene (~ 807 bp) of CalMMV isolated from infected orchid sample was cloned in expression vector, pET-28a ( +) that yielded ~ 31 kDa fusion protein with Histidine tag (His6BP). The expression of fusion CP was confirmed through SDS-PAGE and Western blotting. The His6BP-CalMMV-CP obtained in soluble state after purification was used to immunize New Zealand white rabbit for the production of polyclonal antibodies (PAb). The PAb produced against the purified fusion protein successfully detected CAlMMV in the orchid samples at a dilution of 1:2000 in direct antigen-coated enzyme-linked immunosorbent assay (DAC-ELISA). This study presents the first report of Histidine tag (His6BP) fusion CalMMV-CP-based antibody production and its successful application in the identification of the virus in orchid plants. Outcome of this study will be helpful in routine certification programmes, screening of orchid germplasm and production of CalMMV-free planting materials of orchids.

Atmospheric deposition of mineral dust and associated nutrients over the Equatorial Indian Ocean.

Aerosols are potential supplier of nutrients to the surface water of oceans and can impact biogeochemical processes particularly in the remote locations. The nutrient data from atmospheric supply is poorly reported from the Indian Ocean region. In this study, we present atmospheric nutrients such as reactive nitrogen species (Nitrate, Ammonium, Organic nitrogen), micro-nutrients (e.g. Fe, Mn and Cu) concentration along with mineral dust in the aerosol samples collected over meridional transect during summer (April-May 2018) and monsoon (June-July 2019) months. A significant spatial variation of dust was observed during summer (0.6-22.8 µg m-3) and monsoon (2.8-25.1 µg m-3) months with a decreasing trend from north to south. Dust as well as other nutrient species shows a general north to south decreasing trend, however, no such trend was seen in the soluble trace elements (TEs) concentration. Anthropogenic species like NH4+ and nss-K+ were found below detection limit during monsoon campaign. The fractional solubility (in percentage) of Fe, Mn and Cu were estimated by measuring their concentration in ultrapure water leach which averaged around 0.99 ± 1.12, 31.0 ± 14.9 and 31.1 ± 25.4, respectively during summer and 0.09 ± 0.08, 6.0 ± 8.9, 16.7 ± 9.6, respectively, during monsoon period. Correlation of soluble Fe with total Fe and total acidic species suggest varying dust sources is an important controlling factor for the fraction solubility of Fe with negligible contribution from the chemical processing. However, a significant correlation was observed between total acid and fractional solubility of Mn and Cu suggest role of chemical processing in enhancement of their solubility. Dry deposition flux of aeolian dust was estimated for both campaign using Al concentration and relatively higher fluxes were observed for summer (12.6 ± 8.4 mg·m-2·d-1) and monsoon (8.7 ± 8.4 mg·m-2·d-1) months as compared to model based estimates reported in the literature. Contrastingly, estimated deposition flux of soluble Fe from both campaign displays relatively lower values as compared to model based results which underscores a need for re-evaluation of biogeochemical models with real-time data.

Identification and physico-chemical characterization of microplastics in marine aerosols over the northeast Arabian Sea.

Microplastics (MPs) in the atmosphere can undergo long-range transport from emission regions to pristine terrestrial and oceanic ecosystems. Due to their inherent toxic and hazardous characteristics, MPs pose serious risks to both human well-being and the equilibrium of ecosystem. The present study outlines the comprehensive characterization, spanning physical and chemical attributes of MPs associated with atmospheric aerosols. Total suspended particulates (TSPs) were collected on a quartz fibre filter by operating a high-volume sampler for 24 h during distinct years (March, 2016 and November, 2020) at a coastal location in the northeast Arabian Sea. Subsequent to the sampling, a series of techniques were applied including density separation. The assessment and scrutiny of the MPs was carried out using stereo-zoom microscopy with supplementary validation using advanced fluorescence microscopy for enhanced precision in identification. Our comparative assessment suggests peroxide treatment followed by density separation could be a robust procedure for the definitive identification and characterization of MPs in the atmosphere. Average total abundance of MPs was found to be 1.30 ± 0.14 n/m3 in 2016 and 1.46 ± 0.12 n/m3 in 2020 with fibres, fragments and films having similar relative contributions (41 %, 31 %, 28 % in 2016 and 40 %, 35 %, 25 % in 2020). Fibres were found to be dominant morphotype followed by fragments and films over the coastal region of the Arabian Sea. In order to unravel the detailed chemical nature of these MPs, spectral analysis using µ-FTIR was carried out. The outcome of the analysis showed prevailing polymers as polyvinyl chloride and polymethyl methacrylate (50545 %) as dominant polymers followed by polyester (15 %), styrene butyl methacrylate (11 %), and polyacetal (9 %). MPs present in the vicinity of the Arabian Sea have potential to supply nutrients and toxicants, consequently can contribute to the modulation of the surface water biogeochemical processes.

Evidence for the natural infection of cucurbit chlorotic yellows virus (CCYV) in lettuce plants from India.

In the years 2021 and 2022, lettuce plants showing blistering, chlorosis, mosaic, rosetting/ excess proliferation, and stunting symptoms were subjected to leaf-dip transmission electron microscopy, RT-PCR followed by sequence analysis and bio-assay to unfold the identity of associated virus(es). The association of long filamentous virions (~ 850 nm in length) as seen through leaf-dip transmission electron microscopy suggested the possible infection by a potyvirus or crinivirus, either singly or in combination. RT-PCR assays using generic primers targeting the RdRp region of criniviruses and the NIb region of potyviruses revealed the association of both a crinivirus as well as a potyvirus. The gel-purified RT-PCR products derived from the RdRp region of criniviruses upon cloning, sequencing, and NCBI BLAST analysis indicated the associated crinivirus as cucurbit chlorotic yellows virus (CCYV). Further, RT-PCR assays using specific primers targeting CP and CP minor genes of CCYV followed by cloning and sequencing confirmed its association with the diseased lettuce plants. Besides, the bioassay based on whitefly-mediated virus transmission followed by RT-PCR confirmed the infectivity of CCYV from diseased to healthy lettuce plants. The results of this study confirmed the natural infection of CCYV in lettuce host for the first time in the world indicating its distribution across the crop families.

Posterior Leaflet of Mitral Valve-Is it Really Tri-scalloped? -A Morphological and Morphometric Study in North Indian Cadaveric Hearts.

Background: Maintaining normal left ventricular geometry and function depends on the mitral valve's normal integrity. Irreparable damage to the mitral valve calls for its replacement using either a valve made up of biological tissue or metal, pyrolytic carbon, and similar materials. Materials and Methods: The material consists of 50 formalin-fixed adults, seemingly normal cadaveric hearts of either sex which were received from the Department of Anatomy of various institutes in the north region. These hearts were cut open to access the mitral valve in the left ventricle. Results: In this study, the posterior leaflet was semi-oval in shape being 3.72 cm wide at the base. Usually said to be tri-scalloped, interestingly, it was found so only in 56% of the hearts; being bi-scalloped in 20% and single-cusped in 16% of the hearts. Even four scallops and six scallops were observed in three (6%) and one (2%) hearts, respectively. Conclusions: To conclude, notable variation has been seen in the scallops of posterolateral cusps in the present study. The number of scallops varies greatly as single, double, three, four, or tetra-scalloped and most significant six or hexa-scalloped which has never been reported in the previous studies. To understand the rationale behind each unique architectural layout, such noticeable variations are crucial for scientists around the world. Cardiothoracic surgeons could find this information valuable for mitral valve surgery repair.

Concurrent Cu(II)-initiated Fenton-like reaction and glutathione depletion to escalate chemodynamic therapy.

Chemodynamic therapy is an evolving therapeutic strategy but there are certain limitations associated with its treatment. Herein, we present de novo synthesis and mechanistic evaluation of HL1-HL8 ligands and their corresponding CuII(L1)2-CuII(L8)2. The most active Cu(L2)2 (IC50 = 5.3 µM, MCF-7) complex exclusively depletes glutathione while simultaneously promoting ROS production. Cu(L2)2 also affects other macromolecules like the mitochondrial membrane and DNA while activating the unfolded protein response cascade.

Structural and functional analysis of a novel galactose-binding lectin derived from Chlorella sorokiniana MW769776.

A freshwater green microalgal strain was isolated and the lectin was identified in it by a strong hemagglutination activity (HA) assay. Characterization of the algal strain was found to be Chlorella sorokiniana (MW769776). A single step affinity chromatographic technique was developed to purify Chlorella sorokiniana lectin (CSL) using guar gum as the affinity matrix. The precipitate showed a single active peak with a titer value of 1024 HU, with a concentration of 1111 U, and a purification fold of 9. The purified protein exhibited a single band in SDS-PAGE with a molecular weight of 16 kDa. Analysis by liquid chromatography-electrospray ionization-quadrupole-time of flight mass spectrometry (LC-ESI-Q-TOF-MS) of tryptic-digested purified lectin showed that it was a monomeric protein. A multiple sequence alignment analysis revealed that the peptide sequences of CSL exhibited similarity with the H-type lectin domain of Micractinium conductrix. The structure of CSL was studied by FTIR and homology modeling methods, indicating the presence of α-helix as well as ß-sheet in its secondary structure. Whereas the 3D structure exhibited the similarity with the core protein of light-harvesting reaction center complex of photosystem I. The significance of this study suggests that the characteristics of CSL are consistent with its identification as a hemagglutinin, a type of novel lectin, which suggests its candidature for various biological purposes.Communicated by Ramaswamy H. Sarma.

Forensic analysis of fabricated long bone fractures for gain.

BACKGROUND: Self-inflicting injury is a recognised psychiatric disorder. Such cases are regularly seen in the emergency department, with a false history of assault. Most of these are easily detected by a forensic medicine expert as they present as described in literature but are difficult to explain if altered by professionals for nefarious reasons. METHOD: This was a prospective study carried out at a tertiary centre. The data was collected and analysed from the office records of medico-legal reports prepared at our institute between 25 February 2022 and 25 February 2023. RESULTS: 21 cases were recorded, that met with our criteria. Most involved males (90.5%). Minimum age was 27 years and maximum age 66 years with a mean of 48.6 years and a standard deviation of ±11 years. Most offenders were unemployed (38.1%). Most fractured bone was tibia (47.6%). Left-sided fractures were more common (61.9%). Injuries corresponded with the findings on clothes in three cases (14.3%). DISCUSSION: Fabricated wounds will reflect the intentions of the fabricator and may range from superficial wounds to grievous injury. Only a critical analysis of all medico-legal cases will identify them, and they will have similarities of presentation. CONCLUSION: Such findings have rarely been reported in other parts of India. A diligently prepared medico-legal report and profiling of all cases can help establish patterns of such injuries.

Analytical Study of Mandible: Prerequisite for Sex Determination.

Background: Human skull consists of various bones. One of them is mandible which is quite resistant, tough and shows systemic differences in form between individuals of different sex. It resists putrefaction also. There are characteristic features in the mandible that help us to differentiate sex in case of unknown victims like in mass disasters or in case fragmentary remains of the skeleton are found. Analysis of mandible with regard to its features is of great assistance in the determination of sex. Materials and Methods: A total of 80 dry mandible bones were collected. Morphological and morphometric parameters were studied to determine their sex. A total of nine parameters, i.e., three non-metric and six metric parameters were observed for each mandible. Data was collected for each parameter. Results: Among 80 dry mandible bones, 55 were males and 25 were females. 81.2% males bones had a square chin whereas, 80% females had a rounded chin. Gonial flare was everted in 89% males and inverted in 68% females. Conclusion: Mandible exhibits significant sexual differences. Various morphological and morphometric parameters are essential for sex determination in case of mandible bone.

Determinants of blood pressure control among hypertensive patients of rural areas in a South Indian State: A community based cross sectional study.

Introduction: Optimum blood pressure (BP) control is essential to prevent complications and improve the quality of life of patients with hypertension. This study aimed to explore the determinants of BP control in patients with hypertension on treatment without any other comorbidities. Method: The study was conducted among 429 patients with essential hypertension aged > 30 years residing in four villages of a taluk/tahsil in a South Indian state. A pre-tested semi-structured questionnaire was used to collect data on socio-demographic characteristics, diagnosis and treatment of hypertension, lifestyle factors (e.g. diet and substance use), facilitators and barriers of BP control and anthropometric measurements. BP was measured using World Health Organization standards and classified using the Joint National Committee 8 Guidelines. Descriptive statistics were measured in terms of numbers and percentages. Univariate and multivariate logistic regression analyses were used to determine the significant determinants of BP control. Results: Approximately 64.3% of the participants had their BP under control. The participants aged 46-59 years were more likely to have uncontrolled BP than those aged ≥60 years. The participants with <80% adherence to medication (non-adherent) had a five fold higher odds of having uncontrolled BP than those with >80% adherence. Conclusion: Adherence to medication was the only significant factor for BP control in the present study. Hence, adherence to medication should be addressed with interventions targeted to improve BP control in patients with hypertension.

Prebiotics: Ignored player in the fight against cancer.

BACKGROUND: Prebiotics is a relatively neglected area in cancer research, despite evidence suggesting that it plays a key role in suppressing tumour growth and improving immune function. RECENT FINDINGS: Including prebiotics in the diet has been shown to strengthen the immune system and can better slow down or prevent the growth of tumours. It has also been strongly indicated in various scientific studies that prebiotics can contribute to the sustenance of a healthy microbiome, which in turn plays an important role in increasing the effectiveness and reducing the side effects of cancer treatments. CONCLUSION: In the present review article we highlight the mechanisms by which prebiotics like inulin, fructooligosaccharide (FOS), ß-glucan, pectin, and xylooligosaccharide (XOS) function. Furthermore, the beneficial effect of incorporating prebiotics during cancer therapy to improvise gut health and prevent/reverse the damage caused to patients due to chemotherapy has also been elaborated.

Molecular characterization and SNP identification using genotyping-by-sequencing in high-yielding mutants of proso millet.

Proso millet (Panicummiliaceum L.) is a short-duration C4 crop that is drought tolerant and nutritionally rich and can grow well in marginal lands. Though the crop has many climate-resilient traits like tolerance to drought and heat, its yield is lower than that of common cereals like rice, wheat, and maize. Being an underutilized crop, the molecular resources in the crop are limited. The main aim of the present study was to develop and characterize contrasting mutants for yield and generate functional genomic information for the trait in proso millet. Gamma irradiation-induced mutant population was screened to identify high-yielding mutants, which were evaluated up to M4 generation. One mutant with a dense panicle and high yield (ATL_hy) and one with a lax panicle and low yield (ATL_ly) along with the wild type were sequenced using the genotyping-by-sequencing approach. The variants detected as single nucleotide polymorphisms (SNPs) and insertions-deletions (InDels) were annotated against the reference genome of proso millet. Bioinformatic analyses using the National Center for Biotechnology Information (NCBI) and UniProt databases were performed to elucidate genetic information related to the SNP variations. A total of 25,901, 30,335, and 31,488 SNPs, respectively, were detected in the wild type, ATL_hy mutants, and ATL_ly mutants. The total number of functional SNPs identified in high-yielding and low-yielding mutants was 84 and 171, respectively. Two functional SNPs in the high-yielding mutant (ATL_hy) and one in the low-yielding mutant (ATL_ly) corresponded to the gene coding for "E3 ubiquitin-protein ligase UPL7". Pathway mapping of the functional SNPs identified that two SNPs in ATL_ly were involved in the starch biosynthetic pathway coding for the starch synthase enzyme. This information can be further used in identifying genes responsible for various metabolic processes in proso millet and in designing useful genetic markers.

Geochemical tracing of synoptic scale modern dust transport over the Northeast Arabian Sea during the southwest monsoon.

During the Southwest monsoon (SWM), aeolian dust is mainly supplied via wet deposition over the northeast Arabian Sea (NEAS). To understand their impact on the biogeochemistry of the Arabian Sea, it is important to identify their sources and characteristics. In this context, wet deposit particulate (WDP) samples were collected at a coastal station (Goa; 15.4° N, 73.8° E) in the NEAS during the SWM for three years. These samples were used to characterize and identify mineral dust sources using mineralogical, elemental, and isotopic (Sr and Nd) signatures. The WDP samples were classified as Beginning of Monsoon (BM, June samples), Mid Monsoon (MM, July-August samples) and End of Monsoon (EM, September samples). Clay mineralogical composition indicate high palygorskite content during BM, which subsequently found to decrease in MM, and almost negligible in EM. However, smectite is highest during MM, with moderate presence of palygorskite during this period. The considerable variation in the relative percentages of clay minerals suggests significant temporal variability in dust sources which is further corroborated by the radiogenic isotopic composition. A strong seasonality in the isotopic composition is observed with 87Sr/86Sr ratio being relatively less radiogenic during MM than the BM and highly radiogenic at the EM. Whereas ƐNd values show an opposite trend to 87Sr/86Sr ratios throughout the monsoon, with more radiogenic ƐNd in the MM, and less radiogenic at the EM. End member mixing plot indicate dominant contribution of dust from the Arabian Peninsula (ARB) and Northeast African (NEA) sources during BM and MM, while a shift towards the Thar desert and Southwest Asian (SWA) sources at the EM. Trace elements associated with different sources were quantified and suggest high Fe concentration is associated with NEA dust sources, despite ARB being major supplier of aeolian dust to the Arabian Sea.

Development and application of crude sap-based recombinase polymerase amplification assay for the detection and occurrence of grapevine geminivirus A in Indian grapevine cultivars.

Geminiviruses are known to infect several fields and horticultural crops around the globe. Grapevine geminivirus A (GGVA) was reported in the United States in 2017, and since then, it has been reported in several countries. The complete genome recovered through high-throughput sequencing (HTS)-based virome analysis in Indian grapevine cultivars had all of the six open reading frames (ORFs) and a conserved nonanucleotide sequence 5'-TAATATTAC-3' similar to all other geminiviruses. Recombinase polymerase amplification (RPA), an isothermal amplification technique, was developed for the detection of GGVA in grapevine samples employing crude sap lysed in 0.5 M NaOH solution and compared with purified DNA/cDNA as a template. One of the key advantages of this assay is that it does not require any purification or isolation of the viral DNA and can be performed in a wide range of temperatures (18°C-46°C) and periods (10-40 min), which makes it a rapid and cost-effective method for the detection of GGVA in grapevine. The developed assay has a sensitivity up to 0.1 fg µl-1 using crude plant sap as a template and detected GGVA in several grapevine cultivars of a major grapevine-growing area. Because of its simplicity and rapidity, it can be replicated for other DNA viruses infecting grapevine and will be a very useful technique for certification and surveillance in different grapevine-growing regions of the country.