Biochemical characterization of a novel thermo-halo-tolerant GH5 endoglucanase from a thermal spring metagenome.

A novel endoglucanase gene, celM , was cloned from a thermal spring metagenome. The gene was expressed in Escherichia coli, and the protein was extracted and purified. The protein catalyzed the hydrolysis of amorphous cellulose in a wide range of temperatures, 30-95°C, with optimal activity at 80°C. It was able to tolerate high temperature (80°C) with a half-life of 8 h. Its activity was eminent in a wide pH range of 3.0-11.0, with the highest activity at pH 6.0. The enzyme was tested for halostability. Any significant loss was not recorded in the activity of CelM after the exposure to salinity (3 M NaCl) for 30 days. Furthermore, CelM displayed a substantial resistance toward metal ions, denaturant, reducing agent, organic solvent, and non-ionic surfactants. The amorphous cellulose, treated with CelM , was randomly cleaved, generating cello-oligosaccharides of 2-5 degree of polymerization. Furthermore, CelM was demonstrated to catalyze the hydrolysis of cellulose fraction in the delignified biomass samples, for example, sweet sorghum bagasse, rice straw, and corncob, into cello-oligosaccharides. Given that CelM is a thermo-halo-tolerant GH5 endoglucanase, with resistance to detergents and organic solvent, the biocatalyst could be of potential usefulness for a variety of industrial applications.

Characterization of a novel xylanase from an extreme temperature hot spring metagenome for xylooligosaccharide production.

In this study, the metagenomic resource generated from an aquatic habitat of extreme temperature was screened for the identification of a novel xylanase, XynM1. Gene sequence analysis designated it as a member of glycoside hydrolase (GH) family 10. The metagenomic DNA fragment was cloned, expressed in Escherichia coli, and the purified protein was biochemically characterized. The optimum temperature and pH for the XynM1 xylanase were found to be at 80 °C and 7, respectively. It exhibited worthwhile pH stability by retaining about 70% activity in the range of pH 6 to 9 after the exposure for 12 h at 25 °C. Thermostability analysis established considerable heat tolerance in XynM1 protein at elevated temperatures, displaying about 50% residual activity after the exposure of 40 °C, 50 °C, 60 °C, and 70 °C for 20 h, 12 h, 6 h, and 1.5 h, respectively. The effects of additives such as metals, surfactants, and organic solvents were evaluated on the activity of XynM1. It was able to retain about 50% of its initial activity in the presence of NaCl concentration of 1 to 5 M. The novel xylanase was capable of hydrolyzing the hemicellulosic polymer, derived from diverse biomass sources, e.g., beechwood xylan, wheat arabinoxylan, corncob xylan, and sweet sorghum xylan. The XynM1-treated beechwood xylan manifested catalytic release of xylooligosaccharides (XOS) of 2-6 DP. The novel GH10 xylanase is a promising biocatalyst that could be ascribed for biomass conversion and production of prebiotic XOS biomolecules.

Bioprospecting CAZymes repertoire of Aspergillus fumigatus for eco-friendly value-added transformations of agro-forest biomass.

BACKGROUND: Valorizing waste residues is crucial to reaching sustainable development goals and shifting from a linear fossil-based economy to a circular economy. Fungal cell factories, due to their versatility and robustness, are instrumental in driving the bio-transformation of waste residues. The present work isolated a potent strain, i.e., Aspergillus fumigatus (ZS_AF), from an ancient Zloty Stok gold mine, which showcased distinctive capabilities for efficient hydrolytic enzyme production from lignocellulosic wastes. RESULTS: The present study optimized hydrolytic enzyme production (cellulases, xylanases, and ß-glucosidases) from pine sawdust (PSD) via solid-state fermentation using Aspergillus fumigatus (ZS_AF). The optimization, using response surface methodology (RSM), produced a twofold increase with maximal yields of 119.41 IU/gds for CMCase, 1232.23 IU/gds for xylanase, 63.19 IU/gds for ß-glucosidase, and 31.08 IU/gds for FPase. The secretome profiling validated the pivotal role of carbohydrate-active enzymes (CAZymes) and auxiliary enzymes in biomass valorization. A total of 77% of carbohydrate-active enzymes (CAZymes) were constituted by glycoside hydrolases (66%), carbohydrate esterases (9%), auxiliary activities (3%), and polysaccharide lyases (3%). The saccharification of pretreated wheat straw and PSD generated high reducing sugar yields of 675.36 mg/g and 410.15 mg/g, respectively. CONCLUSION: These findings highlight the significance of an efficient, synergistic, and cost-effective arsenal of fungal enzymes for lignocellulosic waste valorization and their potential to contribute to waste-to-wealth creation through solid-waste management. The utilization of Aspergillus fumigatus (ZS_AF) from an unconventional origin and optimization strategies embodies an innovative approach that holds the potential to propel current waste valorization methods forward, directing the paradigm toward improved efficiency and sustainability.

Preliminary data on cytotoxicity and functional group assessment of a herb-mineral combination against colorectal carcinoma cell line.

OBJECTIVES: The invasive screening methods and the late stage diagnosis of colorectal carcinoma (CRC) are contributing for the devastative prognosis. The gradual shift of the disease pattern among younger generations requires the implementation of phytochemicals and traditional medicines. Arkeshwara rasa (AR) is a herb-mineral combination of Tamra bhasma/incinerated copper ashes and Dwigun Kajjali/mercury sulphide levigated with Calotropis procera leaf juice, Plumbago zeylanica root decoction and the decoction of three myrobalans (Terminalia chebula, Terminalia bellerica, Emblica Officinalis decoction)/Triphala decoction. METHODS: The SW-480 cell line was checked for the cytotoxicity and the cell viability criteria with MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) assay. The acridine orange/ethidium bromide (AO/EtBr) assay revealed the depth of apoptosis affected cells in the fluorescent images. The FTIR analysis exhibited the graphical spectrum of functional groups within the compound AR. RESULTS: The IC50 from the 10-7 to 10-3 concentrations against SW-480 cells was 40.4 µg/mL. The staining of AO/EtBr was performed to visualize live and dead cells and it is evident from the result that number of apoptotic cells increases at increasing concentration of AR. The single bond with stretch vibrations of O-H and N-H are more concentrated in the 2,500-3,200 cm-1 and 3,700-4,000 cm-1 of the spectra whereas, the finger print region carries the O-H and S=O type peaks. CONCLUSIONS: The AR shows strong cyto-toxicity against the SW-480 cells by inducing apoptosis. It also modulates cellular metabolism with the involvement of functional groups which antagonizes the strong acids. Moreover, these effects need to be analyzed further based in the in vivo and various in vitro models.

Characterization of lead sulfide obtained from Naga Bhasma.

BACKGROUND: Lead sulfide nanoparticles were manufactured from lead oxide using a procedure described in the Ayurveda formulary of India, which involved using a quantum of the heat of up to 60 puta, which is officially known as the Shasti puta Naga Bhasma. OBJECTIVE: The study shows sulfurization of nanoparticles decreased their toxicity due to the lower solubility. MATERIALS AND METHODS: The present work used the arsenic sulfide media and traditional puta for processing and the characterization of the same has been conducted. Different analytical techniques like X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-Ray (EDX) spectroscopy, Fourier transform infrared spectroscopy (FTIR), and thermo-gravimetry analysis (TGA) were used. RESULTS: Powder x-ray diffraction, scanning electron microscopy, Fourier transform infrared spectroscopy, Thermogravimetric analysis, and differential thermal analysis all showed that the produced nanoparticles are lead sulfide nanoparticles with a particle size of an average of 84.60 and the crystalline average size of 69.06 nm. CONCLUSION: The rounded, rod, oval, cubic, and circular morphology of the produced lead sulfide nanoparticles can be seen in the SEM image. The stretching and bending functional groups in the sample were alkanes, alkenes, aromatic hydrocarbons, carboxylic acids, alkyl carboxylic acids, alkyl alpha, beta-unsaturated, aldehydes, ketones, carboxylic acid, aliphatic amines, primary amines, secondary amines, alkyl halides, are studied through the FTIR spectrum.

Pharmaceutical characterization and exploration of Arkeshwara rasa in MDA-MB-231 cells.

BACKGROUND: The diverse specificity mode of cancer treatment targets and chemo resistance demands the necessity of drug entities which can address the devastating dynamicity of the disease. OBJECTIVES: To check the anti-tumour potential of traditional medicine rich in polyherbal components and metal nanoparticle namely Arkeshwara rasa (AR). MATERIAL METHODS: The AR was prepared in a modified version with reference from Rasaratna Samuchaya and characterized using sophisticated instrumental analysis including XRD, SEM-EDAX, TEM, TGA-DSC, and LC-MS and tested against the MDA-MB-231 cell line to screen cell viability and the cytotoxicity with MTT, SRB and the AO assay. RESULTS: XRD pattern shows cubic tetrahedrite structure with Sb, Cu, S peaks and trace elements like Fe, Mg, etc. The particle size of AR ranges between 20 and 30 nm. The TGA points thermal decomposition at 210 °C and the metal sulphide peaks in DSC. LC-MS analysis reveals the components of the formulation more on the flavonoid portion. The IC50 value of MTT and SRB are 25.28 µg/mL and 31.7 µg/mL respectively. The AO colorimeter substantiated the cell viability and the apoptosis figures of the same cell line. The AR exhibits cytotoxicity and reaffirms the apoptosis fraction with SRB assay. CONCLUSIONS: The Hesperidine, Neohesperidin, Rutin components in the phytochemical pool can synergize the anti-tumour potential with either influencing cellular pathways or decreasing chemo resistance to conventional treatment. AR need to be further experimented with reverse transcription, flow cytometry, western blotting, etc.

Genotype-by-genotype interkingdom cross-talk between symbiotic nitrogen fixing Sinorhizobium meliloti strains and Trichoderma species.

In the understanding of the molecular interaction between plants and their microbiome, a key point is to identify simplified models of the microbiome including relevant bacterial and fungal partners which could also be effective in plant growth promotion. Here, as proof-of-concept, we aim to identify the possible molecular interactions between symbiotic nitrogen-fixing rhizobia and soil fungi (Trichoderma spp.), hence shed light on synergistic roles rhizospheric fungi could have in the biology of symbiotic nitrogen fixation bacteria. We selected 4 strains of the model rhizobium Sinorhizobium meliloti and 4 Trichoderma species (T. velutinum, T. tomentosum, T. gamsii and T. harzianum). In an experimental scheme of 4 ×4 strains x species combinations, we investigated the rhizobia physiological and transcriptomic responses elicited by fungal spent media, as well as spent media effects on rhizobia-host legume plant (alfalfa, Medicago sativa L.) symbiosis. Fungal spent media had large effects on rhizobia, specific for each fungal species and rhizobial strains combination, indicating a generalized rhizobia genotype x fungal genotype interaction, including synergistic, neutral and antagonistic effects on alfalfa symbiotic phenotypes. Differential expression of a high number of genes was shown in rhizobia strains with up to 25% of total genes differentially expressed upon treatment of cultures with fungal spent media. Percentages over total genes and type of genes differentially expressed changed according to both fungal species and rhizobial strain. To support the hypothesis of a relevant rhizobia genotype x fungal genotype interaction, a nested Likelihood Ratio Test indicated that the model considering the fungus-rhizobium interaction explained 23.4% of differentially expressed genes. Our results provide insights into molecular interactions involving nitrogen-fixing rhizobia and rhizospheric fungi, highlighting the panoply of genes and genotypic interactions (fungus, rhizobium, host plant) which may concur to plant symbiosis.

Ayurvedic preparations of Raudra Rasa inhibit agonist-mediated platelet activation and restrict thrombogenicity without affecting cell viability.

Ayurveda is considered to be one of the most ancient forms of medicine still practiced. The Ayurvedic preparation Raudra Rasa and its derivatives have been widely employed against cancer since the 12th century, but the effect of these traditional formulations on platelet function and signaling has not previously been examined. Here we demonstrate that Raudra Rasa and its derivatives significantly reduce thrombin-induced integrin activation and granule secretion in platelets, as observed by reduced PAC-1 binding and P-selectin externalization, respectively. These formulations also inhibited thrombin-stimulated phosphatidylserine exposure, mitochondrial reactive oxygen species generation, and mitochondrial transmembrane potential in platelets. Consistent with the above, Raudra Rasa significantly reduced thrombin-induced tyrosine phosphorylation of the platelet proteins, as well as phosphorylation of the enzymes AKT and GSK-3ß. In summary, Raudra Rasa inhibits agonist-mediated platelet activation without affecting cell viability, suggesting it may have therapeutic potential as an anti-platelet/anti-thrombotic agent.

Screening of anti-cancerous potential of classical Raudra rasa and modified Raudra rasa modified with hiraka bhasma (nanodiamond) through FTIR & LC-MS analysis.

OBJECTIVES: Raudra rasa is an ayurvedic medicine explicitly prescribed for the treatment of arbuda (cancer), whereas hiraka bhasma has the potential to promote cancer healing properties. Together, these two medicines provide multifunction benefits. This paper analyses the functional groups of Raudra rasa modified with hiraka bhasma and compares it with the classically prepared raudra rasa. To identify the functional group, organic ligands, and active compounds present in samples of raudra rasa (CRR) and modified raudra rasa with hiraka bhasma (MRR) contributing to cancer alleviation by using Fourier transform infrared spectroscopy (FTIR) & LC-MS analysis. METHODS: Classical raudra rasa (CRR), its ingredients, shadguna kajjali (SK); decoction of Piper betel Linn. (PBD); Amaranthus spinosus Linn. (ASD); Boerhaavia diffusa Linn. (BDD); Piper longum Linn. (PLD); cow urine (GM), & similarly modified raudra rasa (MRR), its ingredients, hiraka bhasma (HB); shadguna rasasindura (SHR); water-soluble extract of Piper betel Linn. (PBE); Amaranthus spinosus Linn. (ASE); Boerhaavia diffusa Linn. (BDE); cow urine ark (GA); Piper Longum Linn. (PLE) were subjected to FTIR and LC-MS analysis. RESULTS: Among all 15 samples studied, maximum numbers of peaks (21) were seen in MRR indicating a greater number of functional groups. Further, in MRR, a maximum peak in the double bond region is suggestive of its higher stability compared to CRR. Both the compound is preliminarily a mixture of the number of functional groups like; fluoro, methyl, amino, hydroxy, nitro, methylamino, carbonyl, and iodo groups, having known anti-proliferative activities. By the FT-IR analysis, the biologically active compounds in aqueous and methanol extract of CRR & MRR were identified that have anti-cancerous compounds. In the present study, a total of 40 major compounds like alkaloids, amino acid, carboxylic acid, Flavonoids, Nucleoside, Nucleotide, phenylpropanoid, Sphingosine, stilbenoid, sugar, phosphate, terpenoids, vitamin from aqueous & methanol extract of CRR & MRR were identified by LC-MS. CONCLUSIONS: This research paper highlights the presence of different functional groups and bioactive compounds known to have anti-cancer activities. Thus, this review suggests future recommendations for the design and development of improved anticancer drugs with higher efficacy.

Colorful Treasure From Agro-Industrial Wastes: A Sustainable Chassis for Microbial Pigment Production.

Colors with their attractive appeal have been an integral part of human lives and the easy cascade of chemical catalysis enables fast, bulk production of these synthetic colorants with low costs. However, the resulting hazardous impacts on the environment and human health has stimulated an interest in natural pigments as a safe and ecologically clean alternative. Amidst sources of natural producers, the microbes with their diversity, ease of all-season production and peculiar bioactivities are attractive entities for industrial production of these marketable natural colorants. Further, in line with circular bioeconomy and environmentally clean technologies, the use of agro-industrial wastes as feedstocks for carrying out the microbial transformations paves way for sustainable and cost-effective production of these valuable secondary metabolites with simultaneous waste management. The present review aims to comprehensively cover the current green workflow of microbial colorant production by encompassing the potency of waste feedstocks and fermentation technologies. The commercially important pigments viz. astaxanthin, prodigiosin, canthaxanthin, lycopene, and ß-carotene produced by native and engineered bacterial, fungal, or yeast strains have been elaborately discussed with their versatile applications in food, pharmaceuticals, textiles, cosmetics, etc. The limitations and their economic viability to meet the future market demands have been envisaged. The most recent advances in various molecular approaches to develop engineered microbiological systems for enhanced pigment production have been included to provide new perspectives to this burgeoning field of research.

Transmission modes of COVID-19 disease pandemic in the light of ancient wisdom of Ayurveda medicine: a review.

Human-to-human transmission of the Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) occurs most often when people are in the incubation stage of the disease or are carriers and have no symptoms. SARS-CoV-2 transmission on different levels showed that the cold and dry climate, hot wind and relative humid wind, maximum wind speed, turbulent wind, intensified recirculating flow were causing transmission higher than factors. The role of environmental factors described in Ayurveda like air, water, soil, season, frequent earthquakes, sunlight exposure, frequent thunderstorm with lightening, and factors from arthropods were revalidated in perspective of COVID-19 transmission. The authors searched the information regarding epidemic diseases in Ayurveda textbooks. Pubmed, Science Direct, Wikipedia, Elsevier, Lancet, and Springer were approached for the latest knowledge relating to SARS-CoV-2 and COVID-19. Google Scholar search engine was thoroughly checked upon for scientific evidence regarding the Ayurveda drugs. Various environmental factors like soil, air, water, frequent earthquake disasters, wildlife animals, aquatic birds, space, inevitable disastrous factors, weather or seasonal hazards, violent thunderstorm with lightning, intermediate hosts, sunlight exposure etc. were considered for their role in the genesis of the disease. The Ayurvedic concepts behind the etiology and development of epidemics are the same as modern epidemiological theories. The mysteries of many aspects of the current pandemic might be deciphered by traditional medicine knowledge and thus adding upon to the integrative medicine concept.

Physico-chemical characterization of kajjali, black sulphide of mercury, with respect to the role of sulfur in its formation and structure.

BACKGROUND: Kajjali is used as a base for Ayurvedic herbo-mineral medicines. It is a combination of mercury with sulfur in varying proportions. The ratio of sulfur (S) added to mercury (Hg) directly relates to the therapeutic efficacy of the compound. OBJECTIVE: To analyze the physico-chemical characteristics of samaguna gandhaka kajjali (Hg: S = 1:1) and shadaguna gandhaka kajjali (Hg: S = 1:6). MATERIALS AND METHODS: X-ray diffraction, scanning electron microscopy, X-ray photoelectron spectroscopy (XPS), Fourier transmission infrared spectroscopy, thermo-gravimetry analysis, and atomic absorption spectroscopy were applied to characterize each type of kajjali. RESULTS: It was found that the particle size of the formed kajjali compound increases with a decrease in the mercury to sulfur ratio. The presence of excess sulfur does not change the surface oxidation states as revealed by the XPS analysis. No trace of mercury has been found in both samaguna gandhaka kajjali (SGK-1) and shadguna gandhaka kajjali (SGK-6), indicating a complete Hg reaction with S. CONCLUSION: Kajjali simulates nanomaterial of the modern era and possesses therapeutic efficacy as mentioned in classical Ayurveda texts. Complete trituration of mercury and sulfur combination ends up with this kajjali formation incorporating the potency of nanotherapeutics.

Characterization and antimicrobial study of Trinakantamani (Amber) Pishti.

Background: Trinakantamani Pishti (TMP) is a cardio-tonic (Hridya), styptic (Rakta Stambhaka), astringent (Kashaya) formulation frequently used in varieties of bleeding disorders such as bloody diarrhea (Raktatisaara), Raktarsha (bleeding piles), and disorders of excessive menstruation (Atyartava). Still, no published data is available regarding its characterization. Aim: To generate a fingerprint for raw and processed TMP using sophisticated instrumental techniques to assess antimicrobial activity of TMP. Materials and methods: Three samples of TMP were prepared using the standard reference method. Characterization of TMP was carried out by Fourier-transform infrared spectroscopy (FTIR), energy dispersive X-ray analysis (EDEX) with scanning electron microscopy, powder X-ray diffraction (XRD). Antibacterial activity was carried out by the well-diffusion method. Results: Analysis by scanning electron microscope revealed maximum particle size <5 µm and <3 µm in the raw sample and TMP, respectively. Minimum particle size in TMP ranges from 1 to 2 µm and 701 nm. EDEX analysis shows carbon and oxygen as major constituents while Na, Mg, Ca, Si, Fe, and S were present in traces. XRD pattern indicates the amorphous nature of the drug, while FTIR analysis reveals the presence of functional groups such as O-H, CO2, C = O, C-N, N-H. Heavy metals, total microbial count, and microbial limit test were found to be under permissible limits. Anti-microbial study against tested pathogens Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium did not show any effect of TMP. Conclusion: The results of EDEX study showed that Pishti samples have the small particle size i.e., 701nm than the raw i.e., 1-2 µm, which may facillitate absobtion of drug into the body. All heavy metals in the samples were within the permissible limit. Carbon, hydrogen and oxygen are the chief elements of drug which confirms similarity to the Amber, Since the present work is the first published literature on characterization and anti-microbial study on TMP, the outcome can be considered as fingerprint for the drug prepared using the mentioned reference method.

Analyzing the experiences of adolescent control rats: Effects of the absence of physical or social stimulation on anxiety-like behaviour are dependent on the test.

The present study was designed to systematically assess the control experience routinely used in our laboratory as part of studies on predator odour stress. Specifically, we examined effects of the physical and social components of this control experience on measures of anxiety-like behaviour in adolescent rats. Adolescent animals are at increased susceptibility to environmental perturbations and have been used for such studies much less often. Long-Evans rats of both sexes were subjected to physical stimulation (Exposed or Unexposed) and social stimulation (Single-Housed or Pair-Housed), resulting in four groups. Exposed rats received six 30-min exposures to an enclosed arena containing an unscented piece of cat collar occurring between adolescence and early adulthood, while Unexposed remained in the home cage. Groups of Exposed and Unexposed animals were housed singly (Single-Housed) from early adolescence to early adulthood or Pair-Housed during this time. Experimental procedures began in adolescence and involved repeated assessment of startle amplitude (measure of anxiety-like behaviour) and prepulse inhibition (PPI; a measure of sensorimotor gating) to gauge the short-term impact of social and/or physical stimulation. All animals were re-paired in adulthood prior to a final startle/PPI session to assess if isolation limited to adolescence could impose long-term effects that were not reversible. We also measured anxiety-like behaviour in adulthood using an extended open field test (EOFT; addition of novel objects to the open field on later days), and the elevated plus maze task (EPM), as well as a sucrose preference test (SPT) to measure anhedonia. An absence of social or physical stimulation resulted in increased startle amplitude and some measures of anxiety-like behaviour in the EOFT, but a reduction in such anxiety-like behaviour in the EPM task. These results suggest common neural substrates for the physical and social experiences. Performance in the SPT was unaltered by any experimental treatments. Sensorimotor gating, as measured by PPI, was increased in the absence of physical stimulation with no short-term effect of isolation, or of re-pairing. These results indicate the importance of considering individual components of the rearing environment of rats, while showing the need to use multiple assays of anxiety-like behaviour.

Toxicity study of Lauha Bhasma (calcined iron) in albino rats.

BACKGROUND: Lauha Bhasma (LB) is a complex herbomineral preparation widely used as an Ayurvedic hematinic agent. It is an effective remedy for chronic fever (jirna jvara), phthisis (ksaya), Breathlessness (svasa) etc., and possesses vitality enhancing (vajikara), strength promoting and anti aging (rasayana) properties. OBJECTIVES: The present work was conducted to establish the safety aspects of the use of Lauha bhasma. SETTING AND DESIGN: LB was prepared by Ayurvedic procedures of purification (sodhana), sun drying (bhanupaka), sthalipaka, followed by repeated calcination (marana) and "nectarization" (amrtikarana). The resultant product was subjected to acute and sub acute toxicity studies. MATERIALS AND METHODS: Acute and subacute toxicity study of LB was conducted in albino rats. Criteria for assessment included ponderal changes, change in biochemical parameters viz., LFT and KFT and hematological parameters. Histopathological studies of different organs including liver, kidney, spleen, testis etc., were also conducted to observe pathological changes if any. RESULTS: In the acute toxicity study, the animal group did not manifest any signs of toxicity and no mortality was observed up to 100 times the therapeutic dose (TD). Significant increase in blood urea (27.83%, P < 0.01), serum creatinine (30.92%, P < 0.05), Aspartate aminotransferase (15.09%, P < 0.05), and serum alkaline phosphatase (27.5%, P < 0.01) was evident in group IV (10 TD). A significant increase in serum total protein (6.04%, P < 0.05) level was observed in group III (5 TD). Histopathological examination of livers in group IV (10 TD) showed mild inflammation in terms of bile stasis, peri-portal hepatic inflammation and sinusoidal congestion; lymphocyte infiltration in kidney and intracellular deposits in the splenic tissue. CONCLUSION: Lauha Bhasma was found to be safe at the therapeutic dose and also at five times the therapeutic dose levels. However, alteration in some of the biochemical and haematological parameters along with histopathological findings were evident at the highest dose level.

Critical review of Ayurvedic Varnya herbs and their tyrosinase inhibition effect.

INTRODUCTION: The aspiration for light skin (fair complexion) is becoming pronounced in a greater number of people in the present times with natural products being more in demand than their synthetic counterparts. Research in the area of skin-lightening agents is an expanding field with the knowledge being updated regularly. In Ayurveda, varnya, raktaprasadana, tvacya are few terms specifying skin lightening with respect to its modern counterpart i.e., Tyrosinase inhibition, the most commonly reported method of skin lightening. AIM: The present review is undertaken for screening twenty herbs from Varnya Mahakasaya, Lodhradi varnya gana, Eladi varna prasadana gana and few varnya formulations to evaluate their probable modes of action through which the skin lightening is effected as per both Ayurveda and biomedical concepts. MATERIALS AND METHODS: Critical review of herbs to show varnya property is compiled from various Ayurvedic texts as well as from multiple articles on the internet to justify their skin lightening property on the basis of data collected. RESULT AND CONCLUSION: All the twenty herbs reviewed are found to act as varnya directly (citation as varnya) or indirectly (alleviation of pitta and rakta) as per Ayurveda and to interfere in melanogenesis pathway through tyrosinase inhibition as per biomedicine. This shows their potential to act as good skin whitening agents. Sunthi being a part of many varnya formulations, is the only herb among all reviewed in the present study found to exhibit tyrosinase inhibition without any Ayurvedic citation of varnya property.

Standardization and quality control parameters for Mukta Bhasma (calcined pearl).

BACKGROUND: Mukta Bhasma (MB) is a traditional Ayurvedic preparation for cough, breathlessness, and eye disorders and is a powerful cardiac tonic, mood elevator, and known to promote strength, intellect, and semen production. OBJECTIVES: The present research work was conducted to generate fingerprint for raw and processed MB for quality assessment and standardization using classical and other techniques. SETTING AND DESIGN: Three samples of MB were prepared by purification (sodhana) of Mukta (pearl) followed by repeated calcinations (Marana). Resultant product was subjected to organoleptic tests and Ayurvedic tests for quality control such as rekhapurnata, varitaratva, and nirdhumatva. MATERIALS AND METHODS: For quality control, physicochemical parameters such as loss on drying, total ash value, acid insoluble ash, specific gravity, pH value, and other tests using techniques such as elemental analysis with energy dispersive X-ray analysis (EDAX), Structural study with powder X-ray diffraction, particle size with scanning electron microscopy (SEM) were carried out on raw Mukta, Sodhita Mukta, and triplicate batches of MB. RESULTS: The study showed that the raw material Mukta was calcium carbonate in aragonite form, which on repeated calcinations was converted into a more stable calcite form. SEM studies revealed that in raw and purified materials the particles were found scattered and unevenly arranged in the range of 718.7-214.7 nm while in final product, uniformly arranged, stable, rod-shaped, and rounded particles with more agglomerates were observed in the range of 279.2-79.93 nm. EDAX analysis revealed calcium as a major ingredient in MB (average 46.32%) which increased gradually in the stages of processing (raw 34.11%, Sodhita 37.5%). CONCLUSION: Quality control parameters have been quantified for fingerprinting of MB prepared using a particular method.

Face avulsion and degloving.

There have been sparse reports in literature of avulsion and degloving injuries of individual areas of face like the nose, eyelids, ear and even mandible. Hemi-facial degloving is extremely rare. We present a case of post-assault degloving of the nose, part of forehead with anterior wall of frontal sinus, entire upper and lower eyelids and the cheek. Proper planning and staging of the surgical procedures and use of local flaps, meticulous and proper alignment of tissues gave us good aesthetic and functional outcome with a satisfied patient.

Pharmaceutical Preparation Of Lauha Bhasma

In the present research paper, the pharmaceutical preparation of Lauha bhasma (calcined iron) is being presented. The various procedures adopted in the preparation of Lauha bhasma includes Samanya Shodhana (common purification process for iron) and Vishesha Shodhana (specific purification process for iron), Lauha marana by Bhanupaka (Heating of iron under sunrays) Sthalipaka (Heating of iron in a vessel) and Puta paka (Incineration of iron). Lauha was finally subjected to the process of Amriikaran (nectarization). The process of Puta paka was undergone in electric muffle furnace (EMF) and was repeated for sixty times each under identical conditions, at the temperature of 7500 C till 35th Puta and thereafter at 7000 C till the end of the Marana process i.e. 60 Puta. During the processing of iron, Triphala kwatha (decoction of three myrobalan) has been used as liquid media for Vishesha Shodhana, Bhanupaka, Sthalipaka and Puta paka. The study showed an increase in weight of Lauha after Bhanupaka and Stahlipaka i.e.,, 216% and 105.3%, respectively. It may be due to addition of solid content of Triphala kwatha. After Puta paka, 63%weight gain was observed in final product which may be attributed to addition of ash from Triphala kwatha. The Bhasma obtained fulfils all criteria and was found safe for oral administration.