HPTLC and GC-MS finger-printing of two potential multifunctional siddha tailams: Mathan and maha megarajanga tailam.

The Siddha system of medicine is an ancient medical lineage that is practiced primarily in the southern part of India. Siddha system of medicine has been in practice for thousands of years with documented evidence dating back to the 6th century BCE. According to siddha system of medicine's basic fundamental principle, the human body is made up of 96 thathuvam (primary components), which encompass physical, physiological, psychological, and intellectual aspects. Medicine (marunthu) is classified as a wide range of internal and external medicines. The major components of its medical formulations include plant parts, minerals and animal products. Various methods were carried out for the purification process to eliminate the toxins. Choornam, Guligai, Tailam, Parpam, Chendooram, Kattu, Pasai and Poochu are the most common medicines used in Siddha system of medicine for the treatment of various diseases. The pathophysiological classification of diseases is elaborated in detail in the classical Siddha literature. Siddha system of medicine plays an important role in protecting people from diseases such as COVID-19 by providing immune-protecting and immune-boosting medicines in today's world. Mathan tailam and maha megarajanga tailam are the two unique preparations used widely for various skin diseases including chronic wounds and burns. Scientific validation of both medicines will help in understanding their effectiveness against a typical wound condition. In the present study physio-chemical and phytochemical, HPTLC, and GC-MS analyses were carried out and discussed in detail on the multifunctional properties exhibited in the patient communities.

Pharmacological evaluation of embelin - chitosan nanoparticles as an antidiabetic agent.

Embelin has been reported to possess variety of pharmacological activities such as androgenic antagonists, antiangiogenic, antibacterial, anticancer, anticonvulsant, antidiabetic, antidepressant, antihelmintic, antifertility, antihyperlipidemic, anti-inflammatory, antimalarial, antimitotic, antiobesity and antioxidant properties. The current research work aimed to study the hypoglycemic effect of embelin-chitosan nanoparticles (ECNPs) diabetic rats provoked by streptozotocin (STZ). Embelin nanoparticles (ENPs) were created by combining chitosan, a natural biopolymer, and glutaric acid, a new cross-linker. STZ 50 mg/kg was given intravenously into Sprague-Dawley rats weighing 250-300 g (75-90 days) to induce experimental diabetes. The antidiabetic efficacy of orally administered ECNPs in diabetic rats developed by STZ was investigated, as well as histological examination. When compared to diabetic control rats, ECNPs (25 mg/kg body weight and 50 mg kg body weight) and standard glibenclamide (10 mg/kg body weight) treated rodents exhibited a remarkable drop in glucose contents. Furthermore, histological research showed that ECNPs-treated rats were harmless up to amount of 25 mg/kg bwt. Thus current investigation reveals that ECNPs have antidiabetic potential and may be beneficial in treating hyperglycemia in people.

Detailed studies on microbial adhesion and degradation of polystyrene foam wastes (PSFW) for clean environment.

The present study explores the efficient management of non-degradable polystyrene foam wastes (PSFW) by transforming into a microbial responsive material for an effective biodegradation process. In brief, the study involves three steps, viz., (i) preparation of citrus fruit peel extract from peel wastes; (ii) dissolution of PSFW using the extract and transform to polystyrene sheet (PSS) and characterization of the sheet formed and (iii) finally the microbial adhesion and biofilm formation on PSS. Results revealed that the maximum yield of the peel extract identified as D-limonene was obtained from Citrus sinensis (8.2 ± 0.06 ml/100 g fresh waste). Characterization studies on PSS suggested that there are appreciable changes in the infrared spectrum, thermo gravimetric analysis, gel permeation chromatography analyses, and contact angle measurements in comparison with PSFW. Observations on significant variations in the glass transition temperature of PSFW (100 °C) and PSS (60 °C); decomposition temperature of PSFW (310.93 °C) and PSS (78.18 °C), and molecular weight distribution changes in PSFW (2.00 Mw/Mn) and PSS (1.03 Mw/Mn) suggested the occurrence of structural and molecular changes in PSS. Studies on microbial adhesion and biofilm formation on PSS suggested that amongst six microbial species isolated from the waste dump yard, WD03 strain identified as Lysinibacillus sp., displayed a maximum adhesion and biofilm formation on the surface of the PSS as evidenced through biofilm characterizations, SEM, and fluorescence microscopic analyses respectively. In conclusion, the transformation of PSFW to PSS and the appreciable microbial adhesion and biofilm formation suggested the possibility of the effective management of white nuisance (PSFW) wastes in the environment.

Asperyellone prevents HDF cells from UVB irradiation damages: An elaborated study.

The current study explores the photo-protective effect of asperyellone (AY) (a fungal secondary metabolite), assessed under in vitro condition using human dermal fibroblast cell line. AY was isolated from Aspergillus sp. during the resting phase and purified. The initial cytocompatibility assessment on concentrations of AY and the duration of exposure of UVB irradiations were studied respectively. N-acetyl-L-cysteine (NAC) was used as positive control. Cells were then pretreated with optimized concentration of AY (2.0 µM) and NAC (1 mM) for 1 hour and then UVB irradiated (30 mJ/cm 2 ) for the period of 10 minutes. Results revealed that reactive oxygen species generated upon UVB irradiation found scavenged by the AY pretreatment at a significant level. Furthermore, an appreciable reduction in apoptotic cell count and DNA damages support the scavenging effect of AY. Assessments on the expression of enzymatic and nonenzymatic antioxidants evidently prove the protective role of AY. The reduced expression levels of inflammatory markers (TNF-α and COX-2), collagen degraders (MMP 2 and MMP 9), apoptotic protein expressions (Bax and Bcl-2), and cell-cycle arrest analyses substantiate the photo-protective effect of AY similar to NAC (positive control). Thus, the observations made in the current study indicate the possible role of AY as a photo-protective agent.

Organically modified clay supported chitosan/hydroxyapatite-zinc oxide nanocomposites with enhanced mechanical and biological properties for the application in bone tissue engineering.

The objective of this study is to design biomimetic organically modified montmorillonite clay (OMMT) supported chitosan/hydroxyapatite-zinc oxide (CTS/HAP-ZnO) nanocomposites (ZnCMH I-III) with improved mechanical and biological properties compared to previously reported CTS/OMMT/HAP composite. Fourier transform infrared spectroscopy, powder X-ray diffraction, scanning electron microscopy and transmission electron microscopy were used to analyze the composition and surface morphology of the prepared nanocomposites. Strong antibacterial properties against both Gram-positive and Gram-negative bacterial strains were established for ZnCMH I-III. pH and blood compatibility study revealed that ZnCMH I-III should be nontoxic to the human body. Cytocompatibility of these nanocomposites with human osteoblastic MG-63 cells was also established. Experimental findings suggest that addition of 5wt% of OMMT into CTS/HAP-ZnO (ZnCMH I) gives the best mechanical strength and water absorption capacity. Addition of 0.1wt% of ZnO nanoparticles into CTS-OMMT-HAP significantly enhanced the tensile strengths of ZnCMH I-III compared to previously reported CTS-OMMT-HAP composite. In absence of OMMT, control sample (ZnCH) also showed reduced tensile strength, antibacterial effect and cytocompatibility with osteoblastic cell compared to ZnCMH I. Considering all of the above-mentioned studies, it can be proposed that ZnCMH I nanocomposite has a great potential to be applied in bone tissue engineering.

Development of bone-like zirconium oxide nanoceramic modified chitosan based porous nanocomposites for biomedical application.

Here, zirconium oxide nanoparticles (ZrO2 NPs) were incorporated for the first time in organic-inorganic hybrid composites containing chitosan, poly(ethylene glycol) and nano-hydroxypatite (CS-PEG-HA) to develop bone-like nanocomposites for bone tissue engineering application. These nanocomposites were characterized by FT-IR, XRD, TEM combined with SAED. SEM images and porosity measurements revealed highly porous structure having pore size of less than 1µm to 10µm. Enhanced water absorption capacity and mechanical strengths were obtained compared to previously reported CS-PEG-HA composite after addition of 0.1-0.3wt% of ZrO2 NPs into these nanocomposites. The mechanical strengths and porosities were similar to that of human spongy bone. Strong antimicrobial effects against gram-negative and gram-positive bacterial strains were also observed. Along with getting low alkalinity pH (7.4) values, similar to the pH of human plasma, hemocompatibility and cytocompatibility with osteoblastic MG-63 cells were also established for these nanocomposites. Addition of 15wt% HA-ZrO2 (having 0.3wt% ZrO2 NPs) into CS-PEG (55:30wt%) composite resulted in greatest mechanical strength, porosity, antimicrobial property and cytocompatibility along with suitable water absorption capacity and compatibility with human pH and blood. Thus, this nanocomposite could serve as a potential candidate to be used for bone tissue engineering.

Multifunctional zirconium oxide doped chitosan based hybrid nanocomposites as bone tissue engineering materials.

This paper reports the development of multifunctional zirconium oxide (ZrO2) doped nancomposites having chitosan (CTS), organically modified montmorillonite (OMMT) and nano-hydroxyapatite (HAP). Formation of these nanocomposites was confirmed by various characterization techniques such as Fourier transform infrared spectroscopy and powder X-ray diffraction. Scanning electron microscopy images revealed uniform distribution of OMMT and nano-HAP-ZrO2 into CTS matrix. Powder XRD study and TEM study revealed that OMMT has partially exfoliated into the polymer matrix. Enhanced mechanical properties in comparison to the reported literature were obtained after the addition of ZrO2 nanoparticle into the nanocomposites. In rheological measurements, CMZH I-III exhibited greater storage modulus (G') than loss modulus (G″). TGA results showed that these nanocomposites are thermally more stable compare to pure CTS film. Strong antibacterial zone of inhibition and the lowest minimum inhibition concentration (MIC) value of these nanocomposites against bacterial strains proved that these materials have the ability to prevent bacterial infection in orthopedic implants. Compatibility of these nanocomposites with pH and blood of human body was established. It was observed from the swelling study that the swelling percentage was increased with decreasing the hydrophobic OMMT content. Human osteoblastic MG-63 cell proliferations were observed on the nanocomposites and cytocompatibility of these nanocomposites was also established. Moreover, addition of 5wt% OMMT and 5wt% nano-HAP-ZrO2 into 90wt% CTS matrix provides maximum tensile strength, storage modulus, aqueous swelling and cytocompatibility along with strong antibacterial effect, pH and erythrocyte compatibility.

Development of biomimetic nanocomposites as bone extracellular matrix for human osteoblastic cells.

Here, we have developed biomimetic nanocomposites containing chitosan, poly(vinyl alcohol) and nano-hydroxyapatite-zinc oxide as bone extracellular matrix for human osteoblastic cells and characterized by Fourier transform infrared spectroscopy, powder X-ray diffraction. Scanning electron microscopy images revealed interconnected macroporous structures. Moreover, in this study, the problem related to fabricating a porous composite with good mechanical strength has been resolved by incorporating 5wt% of nano-hydroxyapatite-zinc oxide into chitosan-poly(vinyl alcohol) matrix; the present composite showed high tensile strength (20.25MPa) while maintaining appreciable porosity (65.25%). These values are similar to human cancellous bone. These nanocomposites also showed superior water uptake, antimicrobial and biodegradable properties than the previously reported results. Compatibility with human blood and pH was observed, indicating nontoxicity of these materials to the human body. Moreover, proliferation of osteoblastic MG-63 cells onto the nanocomposites was also observed without having any negative effect.

Differentiation of human gingival mesenchymal stem cells into neuronal lineages in 3D bioconjugated injectable protein hydrogel construct for the management of neuronal disorder

The success of regeneration attempt is based on an ideal combination of stem cells, scaffolding and growth factors. Tissue constructs help to maintain stem cells in a required area for a desired time. There is a need for easily obtainable cells, potentially autologous stem cells and a biologically acceptable scaffold for use in humans in different difficult situations. This study aims to address these issues utilizing a unique combination of stem cells from gingiva and a hydrogel scaffold, based on a natural product for regenerative application. Human gingival mesenchymal stem cells (HGMSCs) were, with due induction, differentiated to neuronal lineages to overcome the problems associated with birth tissue-related stem cells. The differentiation potential of neuronal lineages was confirmed with suitable specific markers. The properties of mesenchymal stem cells in encapsulated form were observed to be similar to free cells. The encapsulated cells (3D) were then subjected to differentiation into neuronal lineages with suitable inducers, and the morphology and gene expression of transient cells were analyzed. HGMSCs was differentiated into neuronal lineages as both free and encapsulated forms without any significant differences. The presence of Nissl bodies and the neurite outgrowth confirm the differentiation. The advantages of this new combination appear to make it a promising tissue construct for translational application.

Phylogenetic Framework and Biosurfactant Gene Expression Analysis of Marine Bacillus spp. of Eastern Coastal Plain of Tamil Nadu.

The present study emphasizes the diversity assessment of marine Bacillus species with special reference to biosurfactant production, respective gene expression, and discrimination among Bacillus licheniformis and Bacillus subtilis. Among the 200 individual species of eastern coastal plain of Tamil Nadu screened, five biosurfactant producing potential bacterial species with entirely different morphology were selected. Biochemical and 16S rRNA gene sequence analysis suggested that all the said five species belong to Bacillus genera but differ in species levels. Biosurfactant of all the five species fluctuates in greater levels with respect to activity as well as to constituents but showed partial similarity to the commercially available surfactin. The expression of srf gene was realized in all of the five species. However, the sfp gene expression was observed only in three species. In conclusion, both B. licheniformis and B. subtilis demonstrate srf gene; nevertheless, sfp gene was expressed only by Bacillus subtilis.

Microbial surfactant mediated degradation of anthracene in aqueous phase by marine Bacillus licheniformis MTCC 5514.

The present study emphasizes the biosurfactant mediated anthracene degradation by a marine alkaliphile Bacillus licheniformis (MTCC 5514). The isolate, MTCC 5514 degraded >95% of 300 ppm anthracene in an aqueous medium within 22 days and the degradation percentage reduced significantly when the concentration of anthracene increased to above 500 ppm. Naphthalene, naphthalene 2-methyl, phthalic acid and benzene acetic acid are the products of degradation identified based on thin layer chromatography, high performance liquid chromatography, gas chromatography and mass analyses. It has been observed that the degradation is initiated by the biosurfactant of the isolate for solubilization through micellation and then the alkali pH and intra/extra cellular degradative enzymes accomplish the degradation process. Encoding of genes responsible for biosurfactant production (licA3) as well as catabolic reactions (C23O) made with suitable primers designed. The study concludes in situ production of biosurfactant mediates the degradation of anthracene by B. licheniformis.

A multilayered supramolecular self-assembled structure from soybean oil by in situ polymerization and its applications.

The present study emphasizes in situ transformation of soybean oil to self-assembled supramolecular multilayered biopolymer material. The said polymer material was characterized and the entrapment efficacy of both hydrophilic and hydrophobic moieties was studied. In brief, soybean oil at varying concentration was mixed with mineral medium and incubated under agitation (200 rpm) at 37 degrees C for 240 h. Physical observations were made till 240 h and the transformed biopolymer was separated and subjected to physical, chemical and functional characterization. The maximum size of the polymer material was measured as 2 cm in diameter and the cross sectional view displayed the multilayered onion rings like structures. SEM analysis illustrated the presence of multilayered honeycomb channeled structures. Thermal analysis demonstrated the thermal stability (200 degrees C) and high heat enthalpy (1999 J/g). Further, this multilayered assembly was able to entrap both hydrophilic and hydrophobic components simultaneously, suggesting the potential industrial application of this material.

Inhibition of UVB-induced oxidative damage and apoptotic biochemical changes in human lymphocytes by 2,5-dihydroxy-3-undecyl-1,4-benzoquinone (embelin).

PURPOSE: The present study investigates the inhibition of Ultraviolet B (UVB, 290-320 nm) radiation-induced oxidative damage in peripheral blood human lymphocytes by embelin extracted from Embelia ribes. MATERIALS AND METHODS: Embelin was extracted, purified and characterized. Prior to inhibitory assessment, a maximum concentration of embelin that was non-toxic was determined. Six experimental groups, including respective controls were made to assess the inhibitory effect of embelin for the selected concentrations of 10 and 20 µg/ml. For the experimental groups; lymphocytes (1 × 10(6) cells) were pre-treated with the chosen concentration of embelin for a period of 60 min and then exposed to UVB for 30 min. UVB radiation inhibitory effect of embelin assessed by measuring antioxidant and lipid peroxidation levels, deoxyribonucleic acid (DNA) damage, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) at scheduled time points after irradiation. RESULTS: Pre-treatment of lymphocytes with embelin prevents UVB-induced oxidative damage. An increase in antioxidant levels in irradiated cells in the presence of embelin and UV absorbance of embelin could be the reason for the decrease in lipid peroxidation level and prevention of DNA damage by UVB radiation. CONCLUSION: Embelin prevents oxidative stress induced by UVB irradiation via its antioxidant property.

Identification and Discrimination of Methicillin Resistant Staphylococcus aureus Strains Isolated from Burn Wound Sites Using PCR and Authentication with MALDI-TOF-MS.

The present study demonstrates isolation and identification of methicillin resistance Staphylococcus aureus (MRSA) strains in the samples collected from burn patients. About 106 swab samples were collected from burn patients of >40% burn injury and were subjected to isolation using nutrient agar followed by screening using Me Re Sa selective medium agar. A total of 10 isolates with identity to S. aureus were obtained and further authenticated using Polymerase Chain Reaction and matrix assisted laser desorption/ionization time of flight mass spectrometry analysis. Presence of mec A gene and the peak pattern observations suggested seven of the 10 isolates are MRSA. Thus, the present study emphasizes the process of identification of MRSA using two different bio-analytical techniques, which authenticate the presence of MRSA.

CD14 signaling reciprocally controls collagen deposition and turnover to regulate the development of lyme arthritis.

CD14 is a glycosylphosphatidylinositol-anchored protein expressed primarily on myeloid cells (eg, neutrophils, macrophages, and dendritic cells). CD14(-/-) mice infected with Borrelia burgdorferi, the causative agent of Lyme disease, produce more proinflammatory cytokines and present with greater disease and bacterial burden in infected tissues. Recently, we uncovered a novel mechanism whereby CD14(-/-) macrophages mount a hyperinflammatory response, resulting from their inability to be tolerized by B. burgdorferi. Paradoxically, CD14 deficiency is associated with greater bacterial burden despite the presence of highly activated neutrophils and macrophages and elevated levels of cytokines with potent antimicrobial activities. Killing and clearance of Borrelia, especially in the joints, depend on the recruitment of neutrophils. Neutrophils can migrate in response to chemotactic gradients established through the action of gelatinases (eg, matrix metalloproteinase 9), which degrade collagen components of the extracellular matrix to generate tripeptide fragments of proline-glycine-proline. Using a mouse model of Lyme arthritis, we demonstrate that CD14 deficiency leads to decreased activation of matrix metalloproteinase 9, reduced degradation of collagen, and diminished recruitment of neutrophils. This reduction in neutrophil numbers is associated with greater numbers of Borrelia in infected tissues. Variation in the efficiency of neutrophil-mediated clearance of B. burgdorferi may underlie differences in the severity of Lyme arthritis observed in the patient population and suggests avenues for development of adjunctive therapy designed to augment host immunity.

Microbial biosurfactants and hydrolytic enzymes mediates in situ development of stable supra-molecular assemblies in fatty acids released from triglycerides.

The present study demonstrates in situ formation of multilamellar stable vesicles (MLSVs) of fatty acids released during the growth of microorganisms in the presence of triglycerides. Release of lipase during initial phase of growth hydrolyzes the triglycerides and release free fatty acids (mono or diglycerides) and glycerol. By extending the growth and the prevailing composition of media (unspent nutrients, salts, pH of the medium, biosurfactants, fatty acids, glycerol) and agitation transforms free fatty acids to MLSV of both cylindrical and spherical macroscopic structures via micelle formation with in 240h of incubation. Cross-sectional view and SEM analysis of macroscopic structures reveal the existence of continuous multilayering. Thermo-gravimetric analysis illustrates the stability of the vesicles. FT-IR analysis emphasizes the presence of amide linkages, responsible for self-assembly processes. Schematic representation of formation of MLSV demonstrated for further understanding. Additional exploration on MLSV formation in arteries and the relationship between MLSV and in situ plaque formation by the components of blood in the arteries are schematically explained and submitted as supporting information (SI-2).

Biopolymer from microbial assisted in situ hydrolysis of triglycerides and dimerization of fatty acids.

The present study demonstrates biopolymer production by in situ bio-based dimerization of fatty acids by microorganism isolated from marine sediments. Microbial isolate grown in Zobell medium in the presence of triglycerides for the period of 24-240 h at 37 degrees C, hydrolyze the applied triglycerides and sequentially dimerized the hydrolyzed products and subsequently polymerized and transformed to a biopolymer having appreciable adhesive properties. Physical (nature, odour, stickyness and tensile strength), chemical (instrumentation) and biochemical (cell free broth) methods of analyses carried out provided the hypotheses involved in the formation of the product as well as the nature of the product formed. Results revealed, lipolytic enzymes released during initial period of growth and the biosurfactant production during later period, respectively, hydrolyze the applied triglycerides and initiate the dimerization and further accelerated when the incubation period extended. The existence and the non-existence of in situ hydrolysis of various triglycerides followed by dimerization and polymerization and the mechanism of transformation of triglycerides to biopolymer are discussed in detail.

Antimicrobial activity of secondary metabolite from marine isolate, Pseudomonas sp. against Gram positive and negative bacteria including MRSA.

A proteobacterium isolated from coastal region of Chennai, India, produced appreciable secondary metabolite and partial purification of the obtained secondary metabolite demonstrated antimicrobial activity against both Gram positive and negative organisms including MRSA (methicillin resistant Staphylococcus aureus). Identification of the isolate using biochemical tests, 16s rDNA sequence analysis, G+C content and electron microscopy studies revealed, isolate belonged to Pseudomonas genera. Extraction, purification, characterization and antimicrobial activity of secondary metabolite carried out using various standard instrumental techniques suggested that the active fraction was of 272 m/z with a stable fragment of 244 m/z and also displayed stable free radical activity assessed using EPR analysis. This stable free radical activity of secondary metabolite mediated its antimicrobial activity.