Topical antifungal keratitis therapeutic potential of Clitoria ternatea Linn. flower extract: phytochemical profiling, in silico modelling, and in vitro biological activity assessment.

Introduction: Fungal keratitis (FK) poses a severe threat to vision, potentially leading to blindness if not promptly addressed. Clitoria ternatea flower extracts have a history of use in Ayurvedic and Indian traditional medicines, particularly for treating eye ailments. This study investigates the antifungal and antibiofilm effects of Clitoria ternatea flower extracts on the FK clinical isolate Coniochaeta hoffmannii. Structural details and key compound identification were analysed through FTIR and GC-MS. Methods: The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of Clitoria ternatea flower extracts were determined using broth dilution and well plate techniques. Biofilm inhibitory activity was assessed through microscopic evaluation, while anti-irritant and cytotoxic properties were evaluated using CAE-EI and MTT assays. Through GC-MS and FT-IR analysis the compounds dissolved in the extract and their functional group were studied, and their toxicity screening and pharmacokinetic prediction were conducted in silico. Subsequently, compounds with high corneal permeability were further identified, and molecular docking and simulation studies at 150 ns were used to investigate their interactions with fungal virulence factors and human inflammatory proteins. Results and Discussion: At a concentration of 250 µg/mL, the Clitoria ternatea flower extract displayed effective biofilm inhibition. MIC and MFC values were determined as 500 and 1000 µg/mL, respectively. CAE-EI and MTT assays indicated no significant irritant and cytotoxic effects up to a concentration of 3 mg/mL. Compounds like 9,9-dimethoxybicyclo[3.3.1]nonane-2,4-dione showed high corneal permeability with strong and stable interactions with fungal virulence cellobiose dehydrogenase, endo ß 1,4 xylanase, and glucanase, as well as corneal inflammation-associated human TNF-α and Interleukin IL-1b protein targets. The findings indicate that extracts from C. ternatea flowers could be formulated for an effective and safe alternative for developing new topical FK therapeutics.

Spontaneous nanoemulsification of cinnamon essential oil: Formulation, characterization, and antibacterial and antibiofilm activity against fish spoilage caused by Serratia rubidaea BFMO8.

The contemporary food industry's uses of nanoemulsions (NEs) include food processing, effective nutraceutical delivery, the development of functional chemicals, and the synthesis of natural preservatives, such as phytocompounds. Although cinnamon essential oil (CEO) is widely used in the cosmetic, pharmaceutical, and food industries, it is difficult to add to aqueous-based food formulations due to its weak stability and poor water solubility. This study describes the formulation of a CEO nanoemulsion (CEONE) by spontaneous emulsification and evaluates its antibacterial and antibiofilm properties against biofilm-forming Serratia rubidaea BFMO8 isolated from spoiled emperor fish (Lethrinus miniatus). Bacteria causing spoilage in emperor fish were isolated and identified as S. rubidaea using common morphological, cultural, and 16S RNA sequencing methods, and their ability to form biofilms and their susceptibility to CEONE were assessed using biofilm-specific methods. The spontaneous emulsification formulation of CEONE was accomplished using water and Tween 20 surfactant by manipulating organic and aqueous phase interface properties and controlling particle growth by capping surfactant increases. The best emulsification, with highly stable nano-size droplets, was accomplished at 750 rpm and a 1:3 ratio concentration. The stable CEONE droplet size, polydispersity index, and zeta potential values were 204.8 nm, 0.115, and -6.05 mV, respectively. FTIR and high-resolution liquid chromatography-mass spectrometry (HR-LCMS) analyses have revealed carboxyl, carbonyl, and phenol-like primary phytochemical functional groups in CEO and CEONE, which contribute to their antibacterial and antibiofilm properties.

Diet-induced animal model anti-obesity, phytochemical profiling, and in silico analysis of culinary plant gokhru (Pedalium murex L.) mucilage.

Plant-based diets (PBDs) are renowned for managing and developing bioactive chemical inhibitors to combat obesity, a well-known global public health concern. There are currently no published research studies examining the effects of food plant mucilage dietary supplements on animal models of obesity induced by high-fat diets (HFD). The present research investigated the anti-obesity properties of the culinary plant Pedalium murex L. mucilage (PMM) in obese albino male rats models fed HFD. PMM's HR-LCMS phytochemical profiling and in silico evaluation of anti-obesity and drug-likeness using Schrodinger's Glide, QikProp, and GROMACS modules were also investigated. In vivo, anti-obesity model animal rat's daily dietary intake, common blood biochemical parameters, and histological examination of the liver and kidney tissues for the development of macrovesicular and microvesicular steatosis were all performed. Among the 46 Phytochemicals profiled, 7(14)-Bisabolene-2, 3, 10,11tetrol, Moschamine, and N-Feruloyltyramine show prominent anti-obesity activity and drug-like characteristics in silico. Rats given PMM showed significantly lower serum levels of total cholesterol (TC), low-density lipoprotein (LDL), and triglycerides (TGs), increased levels of high-density lipoprotein (HDL), as well as macro-and microvesicular steatosis, lobular inflammation of the liver and kidney tissues. This suggests that PMM is an effective natural anti-obesity therapeutic ingredient or dietary supplement with a high concentration of anti-obesity phytochemicals that mainly satisfies the needs for such natural anti-obesity medicine or a supplement.Communicated by Ramaswamy H. Sarma.

Peptide specific monoclonal antibodies of Leptospiral LigA for acute diagnosis of leptospirosis.

Leptospirosis is underdiagnosed due to low sensitivity, need of specialised equipment, and expensive reagents for serological and molecular diagnosis respectively. Considering the sensitivity, rapidity, inexpensive reagents and collection of clinical samples, the monoclonal antibody based antigen detection method from urine samples has been developed and evaluated. LigA (LK90) based B-cell specific epitopes were predicted and synthesised as peptides for the production of monoclonal antibody. LK90543: SNAQKNQGNA (amino acids: 543 to 552), and LK901110: DHHTQSSYTP (amino acids: 1110 to 1119) with VaxiJen score of 1.3719 and 1.2215, respectively were used. Thirty two and 28 urine samples from confirmed and seronegative healthy human subjects, respectively were included for the evaluation of MAb-based dot blot ELISA. The specificity of the evaluated MAbs, P1B1 and P4W2 were found to be in the range of ~93-96%. Moreover, the MAbs did not show cross-reactivity with other bacterial antigens as confirmed by IgG ELISA, further validating its specificity for leptospiral antigens. These findings suggest that the developed MAb based dot blot ELISA is a simple, rapid performed in less than 8 h, inexpensive with a ICER of $8.7/QALY, and affordable in developing countries and area where laboratory facilities are limited.

Urinary catheter indwelling clinical pathogen biofilm formation, exopolysaccharide characterization and their growth influencing parameters.

Self-reproducing microbial biofilm community mainly involved in the contamination of indwelling medical devices including catheters play a vital role in nosocomial infections. The catheter-associated urinary tract infection (CA-UTI) causative Staphylococcus aureus, Enterobacter faecalis, and Pseudomonas aeruginosa were selectively isolated, their phenotypic as well as genotypic biofilm formation, production and monomeric sugar composition of EPS as well as sugar, salt, pH and temperature influence on their in vitro biofilm formation were determined. From 50 culture positive urinary catheters S. aureus (24%), P. aeruginosa (18%), E. faecalis (14%) and others (44%) were isolated. The performed assays revealed their varying biofilm forming ability. The isolated S. aureus ica, E. faecalis esp, and P. aeruginosa cup A gene sequencing and phylogenetic analysis showed their close branching and genetic relationship. The analyzed sugar, salt, pH, and temperature showed that the degree of CA-UTI isolates biofilm formation is an environmentally sensitive process. EPS monosaccharide HPLC analysis showed the presence of neutral sugars (ng/µl) as follows: glucose (P. aeruginosa: 44.275; E. faecalis: 4.23), lactose (P. aeruginosa: 7.29), mannitol (P. aeruginosa: 2.53; S. aureus: 2.62; E. faecalis: 2.054) and maltose (E. faecalis: 7.0042) revealing species-specific presence and variation. This study may have potential clinical relevance for the easy diagnosis and management of CA-UTI.

TaxKB: a knowledge base for new taxane-related drug discovery.

BACKGROUND: Taxanes are naturally occurring compounds which belong to a powerful group of chemotherapeutic drugs with anticancer properties. Their current use, clinical efficacy, and unique mechanism of action indicate their potentiality for cancer drug discovery and development thereby promising to reduce the high economy associated with cancer worldwide. Extensive research has been carried out on taxanes with the aim to combat issues of drug resistance, side effects, limited natural supply, and also to increase the therapeutic index of these molecules. These efforts have led to the isolation of many naturally occurring compounds belonging to this family (more than 350 different kinds), and the synthesis of semisynthetic analogs of the naturally existing molecules (>500), and has also led to the characterization of many (>1000) of them. A web-based database system on clinically exploitable taxanes, providing a link between the structure and the pharmacological property of these molecules could help to reduce the druggability gap for these molecules. RESULTS: Taxane knowledge base (TaxKB, http://bioinfo.au-kbc.org.in/taxane/Taxkb/), is an online multi-tier relational database that currently holds data on 42 parameters of 250 natural and 503 semisynthetic analogs of taxanes. This database provides researchers with much-needed information necessary for drug development. TaxKB enables the user to search data on the structure, drug-likeness, and physicochemical properties of both natural and synthetic taxanes with a "General Search" option in addition to a "Parameter Specific Search." It displays 2D structure and allows the user to download the 3D structure (a PDB file) of taxanes that can be viewed with any molecular visualization tool. The ultimate aim of TaxKB is to provide information on Absorption, Distribution, Metabolism, and Excretion/Toxicity (ADME/T) as well as data on bioavailability and target interaction properties of candidate anticancer taxanes, ahead of expensive clinical trials. CONCLUSION: This first web-based single-information portal will play a central role and help researchers to move forward in taxane-based cancer drug research.

HDACiDB: a database for histone deacetylase inhibitors.

An histone deacetylase (HDAC) inhibitor database (HDACiDB) was constructed to enable rapid access to data relevant to the development of epigenetic modulators (HDAC inhibitors [HDACi]), helping bring precision cancer medicine a step closer. Thousands of HDACi targeting HDACs are in various stages of development and are being tested in clinical trials as monotherapy and in combination with other cancer agents. Despite the abundance of HDACi, information resources are limited. Tools for in silico experiments on specific HDACi prediction, for designing and analyzing the generated data, as well as custom-made specific tools and interactive databases, are needed. We have developed an HDACiDB that is a composite collection of HDACi and currently comprises 1,445 chemical compounds, including 419 natural and 1,026 synthetic ones having the potential to inhibit histone deacetylation. Most importantly, it will allow application of Lipinski's rule of five drug-likeness and other physicochemical property-based screening of the inhibitors. It also provides easy access to information on their source of origin, molecular properties, drug likeness, as well as bioavailability with relevant references cited. Being the first comprehensive database on HDACi that contains all known natural and synthetic HDACi, the HDACiDB may help to improve our knowledge concerning the mechanisms of actions of available HDACi and enable us to selectively target individual HDAC isoforms and establish a new paradigm for intelligent epigenetic cancer drug design. The database is freely available on the http://hdacidb.bioinfo.au-kbc.org.in/hdacidb/website.

Biosynthesis of silver nanoparticles using Acacia leucophloea extract and their antibacterial activity.

The immense potential of nanobiotechnology makes it an intensely researched field in modern medicine. Green nanomaterial synthesis techniques for medicinal applications are desired because of their biocompatibility and lack of toxic byproducts. We report the toxic byproducts free phytosynthesis of stable silver nanoparticles (AgNPs) using the bark extract of the traditional medicinal plant Acacia leucophloea (Fabaceae). Visual observation, ultraviolet-visible spectroscopy, and transmission electron microscopy (TEM) were used to characterize the synthesized AgNPs. The visible yellow-brown color formation and surface plasmon resonance at 440 nm indicates the biosynthesis of AgNP. The TEM images show polydisperse, mostly spherical AgNP particles of 17-29 nm. Fourier transform infrared spectroscopy revealed that primary amines, aldehyde/ketone, aromatic, azo, and nitro compounds of the A. leucophloea extract may participate in the bioreduction and capping of the formed AgNPs. X-ray diffraction confirmed the crystallinity of the AgNPs. The in vitro agar well diffusion method confirmed the potential antibacterial activity of the plant extract and synthesized AgNPs against the common bacterial pathogens Staphylococcus aureus (MTCC 737), Bacillus cereus (MTCC 1272), Listeria monocytogenes (MTCC 657), and Shigella flexneri (MTCC 1475). This research combines the inherent antimicrobial activity of silver metals with the A. leucophloea extract, yielding antibacterial activity-enhanced AgNPs. This new biomimetic approach using traditional medicinal plant (A. leucophloea) barks to synthesize biocompatible antibacterial AgNPs could easily be scaled up for additional biomedical applications. These polydisperse AgNPs green-synthesized via A. leucophloea bark extract can readily be used in many applications not requiring high uniformity in particle size or shape.

Evaluation of allium and its seasoning on toxigenic, nutritional, and sensorial profiles of groundnut oil.

Mitigation of xerophilic storage fungi-associated aflatoxin threat in culinary oil will be a new technology advantage to food industries. Groundnut oil isolate Aspergillus flavus MTCC 10680 susceptibility to Allium species (A. sativum L., A. cepa L., and A. cepa var. aggregatum) extracts, composition, and in silico confirmation of extract's phytoconstituent aflatoxin synthesis inhibition were determined. The behavior of seasoning carrier medium groundnut oil in the presence of Allium was also determined. All the Allium species extracts exhibited concentration dependent in vitro inhibition on mycelial biomass, radial growth, and toxin elaboration. The gas chromatography-mass spectrometry revealed the presence of 28, 16, and 9 compounds in the extracts of A. sativum, A. cepa, A. cepa var. aggregatum, respectively. The Allium phytocostituents-like hexadecanoic acid, 5-Octanoyl-2,4,6(1H,3H,5H)-pyrimidinetrione, Guanosine, and so on, showed higher binding energy with aflatoxin synthesis key enzyme ver1. Allium seasoning increased the typical nutty odor of the groundnut oil with sweet aroma note as well as intensification of pale yellow color. Allium seasoning exhibited the highest aflatoxin detoxification and aroma development without any nutritional loss. Culinary oil Allium seasoning has anti-aflatoxin and food additive potential for use in food industries.

Anti-biofilm activity of Salvadora persica on cariogenic isolates of Streptococcus mutans: in vitro and molecular docking studies.

Salvadora persica sticks are used for chewing and oral-hygiene measures worldwide. The growth inhibition and anti-biofilm effects of various extracts on cariogenic Streptococcus mutans isolates were evaluated. Biofilm inhibition, gas chromatography-mass spectrometry (GC-MS) analyses for phytochemicals and their possible mode of interaction with biofilm response regulators were revealed using LigandFit docking protocols. All S. persica extracts showed considerable inhibitory activity and the cariogenic S. mutans showed varied susceptibility when compared with controls. The percentage reduction in biofilm inhibition obtained for methanol, ethanol, chloroform, acetone, and aqueous extracts were 87.92%, 85.75%, 72.44%, 61.66% and 58.68%, respectively. GC-MS analyses revealed >28 compounds, of which benzyl (6Z,9Z,12Z)-6,9,12-octadecatrienoate, 3-benzyloxy-1-nitro-butan-2-ol and 1,3-cyclohexane dicarbohydrazide interacted efficiently with the bacterial communication quorum-sensing (QS) regulators Streptococcus OmpP and Staphylococcus Lux proteins. The bioactive, dual-function, anti-biofilm agents in S. persica not only inhibit growth, but also control the colonization and accumulation of caries-causing S. mutans.

Identification of source of faecal pollution of Tirumanimuttar River, Tamilnadu, India using microbial source tracking.

Efficient management of deteriorating water bodies can be achieved by determining the sources of faecal pollution. Resourceful techniques for discrimination of the sources of Escherichia coli in surface water have recently been developed, including the use of river water to facilitate faecal indicator surveillance, identification of sources of faecal contamination and employing relevant management practices to maintain water quality. This study was conducted to employ microbial source tracking (MST) techniques for the determination of the sources of faecal pollution based on a water quality investigation of the physico-chemical characteristics and coliform count point of the Tirumanimuttar River. To accomplish this, an MST library-based antibiotic resistance analysis, serotyping and the genomic tool rep-PCR techniques were applied, and the obtained results were analysed statistically. Among 135 and 70 E. coli isolates present in the library and water samples collected from the river and nearby well water sources, respectively, most showed intrinsic, high or moderate resistance to antibiotics. Isolates from human and pig faecal sources were 92% homologous with the samples from the river, whereas isolates from sewage and dairy cattle showed 89% and 80% homology, respectively. These findings indicated that the Tirumanimuttar River is subjected to stress from anthropogenic activities and runoff contaminated with agricultural and human faecal contamination. The sources of faecal pollution identified in this study may facilitate the monitoring and management of the Tirumanimuttar River.