Promising antifungal agents: A minireview.

In the recent past, prevalence of life threatening fungal diseases have increased rapidly in immune-compromised cases such as acquired immunodeficiency syndrome (AIDS), cancer, organ transplant etc. Side by side, the appearance of drug resistance to the presently available antifungal therapeutics is on a rapid rise. It has become a top priority for the academia and pharmaceutical industries to develop new antifungal agents able to combat this resistance, and at the same time, possess potential broad spectrum of activity and minimum toxicity. An understanding of the pharmacological interactions between antifungal agents and their targets offers opportunities for design of new therapeutics. This review discusses the various methodology of drug design, structure activity relationships (SARs), and mode of action of variety of new antifungal agents.

Novel formulations of Bacillus thuringiensis var. kurstaki: an eco-friendly approach for management of lepidopteran pests.

Bacillus thuringiensis (Bt) and entomopathogenic fungi (EPF) are in use for management of insect pests. Continuous use of Bt can lead to problem of resistance development in insect pests. Hence use of combination formulations (CF) of microbials with diverse modes of action has been attempted to slow down the process of resistance development. Suspension concentrate (SC) formulations of a local strain of Bt var. kurstaki DOR Bt-127 were developed singly and in combination with conidia of the EPF Nomuraea rileyi (Nr) and Beauveria bassiana (Bb). Electron microscopy of Bt + Bb CF treated larvae of Helicoverpa armigera revealed simultaneous infection by both microbials indicating their compatibility. Endotoxin contents in Bt-SC, Bt + Bb and Bt + Nr CFs were 5.0, 4.7 and 4.7%, respectively. These formulations were effective against larvae of Spodoptera litura, H. armigera and Achaea janata coupled with a lowering of the effective requirement of Bt and EPF. In multi-location field trials, Bt-SC and Bt + Nr CF were highly effective against S. litura and A. janata on castor as well as H. armigera and Thysanoplusia orichalcea on sunflower. However, Bt + Bb CF was highly effective only on sunflower against H. armigera and T. orichalcea. All formulations had 24 months shelf-life at room temperature. DOR Bt-127 based SC formulations developed singly and in combination with Nr and Bb were effective against major lepidopteran pests of castor and sunflower and did not lose viability under storage at room temperature. The CFs of Bt with EPF could prove promising for mitigating resistance development to Bt.

Awareness on cancer cervix, willingness, and barriers for screening of cancer cervix among women: A community-based cross-sectional study from urban Pondicherry.

BACKGROUND: Early detection of cervical cancer can significantly reduce the associated morbidities and mortality. However, uptake of screening for cervical cancer in India is not encouraging. OBJECTIVES: To assess the awareness about cervical cancer, willingness, and barriers for undergoing screening of cervical cancer among women in urban Pondicherry. METHODS: This cross-sectional study was conducted among women of 30-65 years in urban Pondicherry during January - July 2019. A total of 219 women, selected using two-stage random sampling, were interviewed using a pretested semi-structured questionnaire. Multistep multivariable logistic regression was done to identify the independent correlates of willingness to undergo screening for cervical cancers. RESULTS: About one-third women were aware of cervical cancer. Awareness was more among women who were young, had higher education, had family history of cancer, and currently working. Awareness of risk factors, signs and symptoms of cancer cervix was low. Although 60% of the women, who have been aware of cervical cancer, were aware of possibility of early detection, <15% were aware of the various methods. 32% of the women were willing to undergo screening for cervical cancer, and occupation, family history of cancer, and knowledge about risk factors were found to be independent correlates. Fear and "not having signs and symptoms" were the major reasons for unwillingness. CONCLUSION: Level of awareness and willingness for undergoing screening of cervical cancer was low in study area. Targeted interventions for awareness and health system efforts for addressing the reasons behind unwillingness are required.

Analysis of the human protein interactome and comparison with yeast, worm and fly interaction datasets.

We present the first analysis of the human proteome with regard to interactions between proteins. We also compare the human interactome with the available interaction datasets from yeast (Saccharomyces cerevisiae), worm (Caenorhabditis elegans) and fly (Drosophila melanogaster). Of >70,000 binary interactions, only 42 were common to human, worm and fly, and only 16 were common to all four datasets. An additional 36 interactions were common to fly and worm but were not observed in humans, although a coimmunoprecipitation assay showed that 9 of the interactions do occur in humans. A re-examination of the connectivity of essential genes in yeast and humans indicated that the available data do not support the presumption that the number of interaction partners can accurately predict whether a gene is essential. Finally, we found that proteins encoded by genes mutated in inherited genetic disorders are likely to interact with proteins known to cause similar disorders, suggesting the existence of disease subnetworks. The human interaction map constructed from our analysis should facilitate an integrative systems biology approach to elucidating the cellular networks that contribute to health and disease states.

Impact of biopolymer-based Trichoderma harzianum seed coating on disease incidence and yield in oilseed crops.

The use of microbe-based biological control for crop pests is recognized as an environmentally safe substitute for conventional chemical pesticides. However, the practical application of microbial inoculants in large-scale agriculture is underexplored, impeding their widespread commercial adoption. This study addresses the scarcity of research on effective delivery methods for microbial inoculants, particularly through seed coating, which has the potential to be a cost- and time-efficient strategy in crop management. In this research, the Trichoderma harzianum strain Th4d, a biological control agent (BCA), was incorporated into specially formulated biopolymeric compositions based on chitosan and cellulose. The efficacy of this seed coating approach was tested against various soil- and seed-borne pathogens in oilseed crops, including soybean, groundnut, and safflower. Results indicate that safflower treated with the biopolymer chitosan-based T. harzianum Th4d 1 % liquid formulation blend exhibited a higher seed yield of 793 kg/ha, seed germination of 84.7 %, and a significant reduction in wilt and root rot by 64.7 %. In groundnut crops, the seed coating led to a seed germination rate of 88.6 %, a 72 % reduction in root rot incidence, and a seed yield of 3040 kg/ha. Similarly, soybean crops treated with the biopolymer chitosan and T. harzianum Th4d displayed 83.4 % seed germination, a 70.9 % reduction in root rot, and a seed yield of 1239 kg/ha. Further on-farm evaluations demonstrated promising results, with the biopolymer chitosan-based T. harzianum Th4d 1 % liquid formulation blend seed treatment showing a high incremental cost-benefit ratio in safflower (1:4.5), soybean (1:2.5), and groundnut crops (1:3.3). This study underscores the potential of microbe-based seed coating as a sustainable and economically viable strategy for pest management in oilseed crops."

Programmable chitosan-based double layer seed coating for biotic and abiotic-stress tolerance in groundnut.

In the face of agricultural challenges posed by both abiotic and biotic stressors, phytopathogens emerge as formidable threats to crop productivity. Conventional methods, involving the use of pesticides and microbes, often lead to unintended consequences. In addressing this issue, ICAR -Indian Institute of Oilseeds Research (ICAR-IIOR) has developed a chitosan-based double-layer seed coating. Emphasizing crop input compatibility, entrapment, and characterization, the study has yielded promising results. The double-layer coating on groundnut seeds enhanced germination and seedling vigor. Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) confirmed the structural changes and entrapment of crop inputs. The persistence of T. harzianum (Th4d) and Bradyrhizobium sp. in chitosan blended film in studied soils revealed that viable propogules of Th4d were recorded in double layer treatment combination with 3.54 and 3.50 Log CFUs/g of soil (colony forming units) and Bradyrhizobium sp. with 5.34 and 5.27 Log CFUs/g of soil at 90 days after application (DAA). Root colonization efficacy studies of Th4d and Bradyrhizobium sp. in groundnut crop in studied soils revealed that, maximum viable colonies were observed at 45 days after sowing (DAS). This comprehensive study highlights the potential of chitosan-based double-layer seed coating providing a promising and sustainable strategy for stress management in agriculture.

An in silico molecular docking, ADMET and molecular dynamics simulations studies of azolyl-2H-chroman-4-ones as potential inhibitors against pathogenic fungi and bacteria.

Antimicrobial resistance is a major global threat. In an attempt to discover new compounds with improved efficiency and to overcome drug resistance, a library of 3960 compounds was designed as conformationally rigid analogues of oxiconazole with 2H-chroman-4-one, azole and substituted phenyl fragments. The antifungal and antibacterial activity of the compounds was evaluated using molecular docking studies in the active site of six fungal and four bacterial proteins to establish the binding affinity of the designed ligands. In-silico ADME and Lipinski's rule were used to establish the drug-likeness properties of the compounds. This study revealed that all the designed compounds had a high binding affinity with the target proteins and formed H-bond and π-π interactions. The identified hits have been subjected to molecular dynamics simulations to study protein-ligand complex stability. This study has led to the identification of important compounds that can be developed further as therapeutic agents against pathogenic fungi and bacteria.Communicated by Ramaswamy H. Sarma.

HIGHLIGHTSVirtual screening of a library of 3960 2H-chroman-4-one derivatives against pathogenic fungi and bacteriaScreened via molecular docking, ADME and Lipinski filterBind to the active site of the protein target with good binding affinityMD simulation shows stability of the protein-ligand complex of identified hits.

A Study of Clinical Manifestations and their Association with Antinuclear Antibodies in Various Autoimmune Connective Tissue Disorders.

Background: Autoimmune connective tissue diseases (AICTD) present with a myriad of clinical manifestations, including cutaneous. These disorders occur because of immune dysregulation that produces autoantibodies targeting connective tissue and internal organs. Screening these autoantibodies not only aids in the diagnosis but also in predicting specific organ involvement and the risk of complications related to the disease. Aims: This study was conducted (a) to study various cutaneous and systemic manifestations of AICTD, (b) to study the antinuclear antibody (ANA) profile and (c) to determine the association between systemic manifestations and antinuclear antibodies. Methodology: Thirty cases of autoimmune connective tissue disease were recruited for the study. A physical examination, clinical profile and ANA profile were done. Results: Nonscarring alopecia (83.3%) was the commonest cutaneous manifestation noted, followed by photosensitivity (73.3%). The most common system affected was musculoskeletal (67%), followed by renal (40%). Anti-dsDNA antibodies were significantly associated with musculoskeletal involvement (85%) with a P value of 0.038 and anti-Sm antibodies with neurological involvement (87%), followed by renal involvement (75%) with a P value of 0.018 and 0.001, respectively. Anti-SCL 70 antibodies were significantly associated with lung involvement (75%), with a P value of 0.009 and the presence of anti-SS-A antibodies with cardiovascular involvement (40%) with a P value of 0.014. Conclusion: Antinuclear antibodies are diagnostic as well as prognostic biomarkers for AICTD and contribute to precision medicine. These antibodies serve as markers to pursue involvement of organs, which in turn helps the treating physician to choose appropriate preventive measures.

Fe and Zn Metal Nanocitrates as Plant Nutrients through Soil Application.

Nanocitrates of iron (Fe) and zinc (Zn) in the form of plant nanonutrients were examined for their behavior in soil and the uptake of these by 20-day old groundnut (Arachis hypogaea) seedlings under greenhouse conditions. The Fe (0.04 to 0.008 mmol/kg of soil) and Zn (0.02 to 0.004 mmol/kg of soil) nanocitrates were applied to soil and compared with commercial counterparts (FeSO4, ZnSO4, nano-Fe, nano-Zn, Fe-EDTA, Zn-EDTA). The combined nanocitrate compositions were also formulated by physical means and characterized. The plant uptake of Fe and Zn was determined through atomic absorption spectrometry (AAS). All the treated plants showed good germination and higher vigor indexes compared to the control treatments. The highest available Fe and Zn soil contents after leaching were 150.5 and 18.9 mg/kg, respectively, in combined nanocitrate compositions, whereas in the control (untreated) soil, the Fe and Zn contents were 6.0 and 0.7 mg/kg, respectively. The plant's Fe content was 0.48 mg/pot for the combined nanocitrate composition, and that of the untreated plant sample was 0.02 mg/pot. The plant's Zn content was 82.3 µg/pot for pure zinc citrate, and the respective untreated-plant Zn content was 2.1 µg/pot. These values are better than those observed for commercial fertilizers. Additionally, no trend in promotional and antagonistic correlations between Fe and Zn in combined nanocitrates was observed in the studied period (20 days in duration). Among the 34 synthesized citrates, six nanocitrates show promising trends for evaluation under field conditions with higher stability.

Tissue Engineering of Cartilage Using Collagen Scaffold Enriched with Plant Polysaccharides.

Degenerative diseases associated with articular cartilage pose a huge burden on health care economics. The nature of the tissue involved and the changes therein do not allow self-healing; and most of these problems are progressive. Tissue engineering offers some solutions provided we focus on the right kind of cells and the appropriate surrounding niches created for a particular tissue. The present study deals with the formation of polysaccharide rich stable scaffold of collagen after cross-linking with oxidized gum arabic. The scaffold was tested for its biocompatibility and ability to support cells. The in vitro cytotoxicity of the scaffolds toward induced pluripotent stem cells and chondrocytes was evaluated. Evaluation of expression of lineage specific markers indicates differentiation of induced pluripotent stem cells to chondrogenic lineage and maintenance of chondrocytes per se when grown in the scaffold. Animal studies were carried out to study the efficacy of the scaffold to repair the knee injuries. Cells along with the scaffold appeared to be the best filling, in repair of injured cartilage. These studies show that these scaffolds are potential candidates in applications such as tissue engineering of cartilage.

Metal citrate nanoparticles: a robust water-soluble plant micronutrient source.

A series of iron (Fe) and zinc (Zn) plant nanonutrients in citrate form were prepared by an eco-friendly solid-state grinding of the respective nitrates and citric acid. Ball-milling of the as-prepared Fe and Zn citrates resulted in nanosize particles. The as-prepared and ball-milled Fe and Zn citrates were characterized using Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis and differential thermal analysis (TGA/DTA), and powder X-ray diffraction (XRD). The particle size and morphology of the obtained samples were studied using a scanning electron microscope (SEM) and transmission electron microscope (TEM). The obtained nanosized Fe and Zn citrates were analyzed for their plant uptake in the test crop soybean (var. JS-335) using the white-sand technique. The concentration of nutrients was estimated by atomic absorption spectrometry (AAS). A significant increase in nutrient absorption was observed in 6 h ball-milled samples of both Fe (789.8 µg per g of dry weight) and Zn (443.8 µg per g of dry weight) citrates. Such an increased nutrient absorption is due to the high mobility of nanocitrates. Therefore, nanocitrates can serve as an excellent source of plant nutrients in agriculture.

Refunctionalization of Decellularized Organ Scaffold of Pancreas by Recellularization: Whole Organ Regeneration into Functional Pancreas.

BACKGROUND: Tissue engineering centers on creating a niche similar to the natural one, with a purpose of developing an organ construct. A natural scaffold can replace none while creating a scaffold unique to each tissue in composition, architecture and cues that regulate the character of cells. METHODS: Whole pancreas from mouse was decellularized using detergent and enzymes, followed by recellularizing with MSC from human placenta. This construct was transplanted in streptozotocin induced diabetic mice. Histopathology of both decellularized and recellularized transplanted pancreas and qPCR analysis were performed to assess its recovery. RESULTS: Decellularization removes the cells leaving behind extracellular matrix rich natural scaffold. After reseeding with mesenchymal stem cells, these cells differentiate into pancreas specific cells. Upon transplantation in streptozotocin induced diabetic mice, this organ was capable of restoring its histomorphology and functioning. Restoration of endocrine (islets), the exocrine region (acinar) and vascular network was seen in transplanted pancreas. The process of functional recovery of endocrine system took about 20 days when the mice start showing blood glucose reduction, though none achieved gluconormalization. CONCLUSION: Natural decellularized scaffolds of soft organs can be refunctionalized using recipient's mesenchymal stem cells to restore structure and function; and counter immune problems arising during transplantation.

A novel chitosan biopolymer based Trichoderma delivery system: Storage stability, persistence and bio efficacy against seed and soil borne diseases of oilseed crops.

Management of seed and soil borne fungal plant pathogens using fungal species belonging to the genus Trichoderma is gaining importance. Seed coating with powder based formulations of Trichoderma is most widely adopted by the researchers and farmers as well. Delivery system that leads to good adherence of fungal propagules on seed surface, minimizing the wastage of active ingredient, sustained and timely release during treatment process is very important for effective season long protection. Chitosan-PEG (Polyethylene glycol) (Cts-PEG) blend containing Trichoderma harzianum (Th4d) (Cts-PEG-Th) spores is developed and its storage stability, persistence in soil and bio efficacy against seed and soil borne pathogens of groundnut and safflower crops is studied. The blend was stable without much changes in pH throughout the storage period. Persistence studies conducted for 3 months revealed that Cts-PEG-Th amended soil, Trichoderma got released from polymer film slowly and reached a maximum of log 8 CFUs by 30 days and there after started declining to retain log 6 CFUs at 90 days. In shelf life study, the chitosan blend was able to maintain Trichoderma counts of log 10.0 and log 10.2 over a period of 6 months at storage temperatures of 30 °C and 4 °C, respectively and the antagonistic activity unaffected against three plant pathogens viz. Macrophomina phaseolina, Fusarium oxysporumf. sp.ricini and Aspergillus niger over a period of 6 months of storage. Bio efficacy testing in germination towels and green house pot studies revealed the effectiveness of seed treatment with Cts-PEG-Th blend significantly increasing the germination and seedling vigour and reducing the diseases in groundnut (peanut) and safflower.

Development of chitosan-PEG blended films using Trichoderma: Enhancement of antimicrobial activity and seed quality.

Fungi under genus Trichoderma as ameliorates of biotic and abiotic stresses in cultivated crops is gaining popularity world-wide and their application in conjunction with seed coating polymers is an attractive proposition to reduce bioagent wastage and harnessing benefits of combined application. The synergistic action of Trichoderma with natural polymers like chitosan can enhance antimicrobial activity. A series of blended film solutions were synthesized by using chitosan, PEG and plasticizer in varying concentrations. The optimization of blended film composition and dose for coating of seeds was done w.r.t seed coating. Studies on compatibility of film forming ingredients with Trichoderma have not shown any inhibition and antimicrobial activity has shown different levels of inhibition of plant pathogens. Films were structurally characterized by XRD, SEM, FT-IR, TGA, DSC. The optimized film solution in combination with different Trichoderma strains improved seed quality parameters in test crop castor (Ricinus communis). Significant increase in vigour index (3110) was observed with Th4d treatment followed by chitosan and Th4d combination formulation (3023). In conclusion, the optimized chitosan-PEG-Th blend was effective in enhancing seed germination and plant growth of castor. The material can be further tested under large field evaluation as a seed coating agent against various plant diseases.

Human protein reference database--2006 update.

Human Protein Reference Database (HPRD) (http://www.hprd.org) was developed to serve as a comprehensive collection of protein features, post-translational modifications (PTMs) and protein-protein interactions. Since the original report, this database has increased to >20 000 proteins entries and has become the largest database for literature-derived protein-protein interactions (>30 000) and PTMs (>8000) for human proteins. We have also introduced several new features in HPRD including: (i) protein isoforms, (ii) enhanced search options, (iii) linking of pathway annotations and (iv) integration of a novel browser, GenProt Viewer (http://www.genprot.org), developed by us that allows integration of genomic and proteomic information. With the continued support and active participation by the biomedical community, we expect HPRD to become a unique source of curated information for the human proteome and spur biomedical discoveries based on integration of genomic, transcriptomic and proteomic data.

[New approaches to the treatment of the HIV-infected patient]. / Nouvelles approches dans la prise en charge de l'infection à VIH.

HIV infection remains a major problem of public health in Belgium as well as globally. The number of new diagnosies of HIV infection in Belgium remains between two and three daily. Given the dramatic effect of antiretroviral therapy on the mortality due to HIV infection, the number of patients is constantly increasing. The different problems related to HIV care are also changing. Aging of the patients and chronic exposure to antiretroviral medications have induced new complications. We will present in this brief article several new experimental and clinical approaches in which our centre has participated during the last two years.

Human protein reference database as a discovery resource for proteomics.

The rapid pace at which genomic and proteomic data is being generated necessitates the development of tools and resources for managing data that allow integration of information from disparate sources. The Human Protein Reference Database (http://www.hprd.org) is a web-based resource based on open source technologies for protein information about several aspects of human proteins including protein-protein interactions, post-translational modifications, enzyme-substrate relationships and disease associations. This information was derived manually by a critical reading of the published literature by expert biologists and through bioinformatics analyses of the protein sequence. This database will assist in biomedical discoveries by serving as a resource of genomic and proteomic information and providing an integrated view of sequence, structure, function and protein networks in health and disease.

Distinct DNA methylation profiles in malignant mesothelioma, lung adenocarcinoma, and non-tumor lung.

DNA methylation markers provide a powerful tool to make diagnoses based on genetic material obtained directly from tumors or from "remote" locations such as sputum, pleural fluid, or serum. In particular when limited cell numbers are available, amplifyable DNA markers can provide a very sensitive tool for cancer detection and classification. Malignant mesothelioma (MM), an aggressive cancer strongly associated with asbestos exposure, can be difficult to distinguish from adenocarcinoma of the lung when limited material is available. In an attempt to identify molecular markers for MM and adenocarcinoma, we examined the DNA methylation status of 14 loci. Analysis of methylation levels in 10 MM and 8 adenocarcinoma cell lines showed that methylation of APC was significantly elevated in adenocarcinoma compared to MM cell lines (P=0.0003), while methylation of CDH1 was higher in MM (P<0.02). Subsequent examination of the methylation status of the 14 loci in 6 MM and 7 adenocarcinoma primary tumors, which yielded similar methylation profiles, supported these observations. Comparison of methylation in MM cell lines and tumors versus non-tumor lung tissue indicated that APC exhibits less methylation in MM (P=0.003) while RASSF1, PGR1, ESR1, and CDH1 show more methylation in MM, the latter two showing the most significant difference between the two tissue types (P< or = 0.0001). Comparison of methylation in adenocarcinoma cell lines and tumors versus non-tumor lung tissue showed methylation of ESR1, PGR1 and RASSF1 to be significantly elevated in adenocarcinoma, with RASSF1 being most significant (P=0.0002). Thus, with the examination of 14 loci, we have identified 5 candidates that show potential for distinguishing between MM, adenocarcinoma and/or non-cancer lung. Our observations support the strong potential of methylation markers as tools for accurate diagnosis of neoplasms in and around the lung.

Plasma Proteome Database as a resource for proteomics research.

Plasma is one of the best studied compartments in the human body and serves as an ideal body fluid for the diagnosis of diseases. This report provides a detailed functional annotation of all the plasma proteins identified to date. In all, gene products encoded by 3778 distinct genes were annotated based on proteins previously published in the literature as plasma proteins and the identification of multiple peptides from proteins under HUPO's Plasma Proteome Project. Our analysis revealed that 51% of these genes encoded more than one protein isoform. All single nucleotide polymorphisms involving protein-coding regions were mapped onto the protein sequences. We found a number of examples of isoform-specific subcellular localization as well as tissue expression. This database is an attempt at comprehensive annotation of a complex subproteome and is available on the web at http://www.plasmaproteomedatabase.org.