Search results for: Bindhu Rani Thumkunta

Authors: Alok Kumar Yadav, Saurabh Saraswat, Preeti Sirohi, Manjoo Rani, Sameer Srivastava, Manish Pratap Singh, Nand K. Singh

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The development of antibiotic resistance in bacteria is increasing at an alarming rate, and this is considered as one of the most serious threats in the history of medicine, and an alternative solution should be derived so as to tackle this problem. In many countries, people use the medicinal plants for the treatment of various diseases as these are cheaper, easily available and least toxic. Syzygium cumini is used for the treatment of various kinds of diseases but their mechanism of action is not reported. The antimicrobial activity of Syzygium cumini was tested by the well diffusion assay and zone of inhibition was reported to be 20.06 mm as compared to control with MIC of 0.3 mg/ml. Genomic DNA fragmentation of Bacillus subtilis revealed apoptosis and FE-SEM indicate cell wall cracking on several intervals of time. Propidium iodide staining results showed that few bacterial cells were stained in the control and population of stained cells increase after exposing them for various period of time. Flow cytometric kinetic data analysis on the membrane permeabilization in bacterial cell showed the significant contribution of antimicrobial potential of the seed extract on antimicrobial-induced permeabilization. Two components of Syzygium cumini methanolic seed extract was found to be quite active against four enzymes like PDB ID- 1W5D, 4OX3, 3MFD and 5E2F which have a very crucial role in membrane synthesis in Bacillus subtilis by in silico analysis. Through in silico analysis, lupeol showed highest binding energy for macromolecule 1W5D and 4OX3 whereas stigmasterol showed the highest binding energy for macromolecule 3MFD and 5E2F respectively. It showed that methanolic seed extract of Syzygium cumini can be used for the inhibition of foodborne infections caused by Bacillus subtilis and also as an alternative of prevalent antibiotics.

Keywords: antibiotics, Bacillus subtilis, inhibition, Syzygium cumini

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Authors: V. P. Muhamed Shajudheen, K. Viswanathan, K. Anitha Rani, A. Uma Maheswari, S. Saravana Kumar

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In this paper, a simple chemical precipitation route for the preparation of titanium dioxide nanoparticles, synthesized by using titanium tetra isopropoxide as a precursor and polyvinyl pyrrolidone (PVP) as a capping agent, is reported. The Differential Scanning Calorimetry (DSC) and Thermo Gravimetric Analysis (TGA) of the samples were recorded and the phase transformation temperature of titanium hydroxide, Ti(OH)₄ to titanium oxide, TiO₂ was investigated. The as-prepared Ti(OH)₄ precipitate was annealed at 800°C to obtain TiO₂ nanoparticles. The thermal, structural, morphological and textural characterizations of the TiO₂ nanoparticle samples were carried out by different techniques such as DSC-TGA, X-Ray Diffraction (XRD), Fourier Transform Infra-Red spectroscopy (FTIR), Micro Raman spectroscopy, UV-Visible absorption spectroscopy (UV-Vis), Photoluminescence spectroscopy (PL) and Field Effect Scanning Electron Microscopy (FESEM) techniques. The as-prepared precipitate was characterized using DSC-TGA and confirmed the mass loss of around 30%. XRD results exhibited no diffraction peaks attributable to anatase phase, for the reaction products, after the solvent removal. The results indicate that the product is purely rutile. The vibrational frequencies of two main absorption bands of prepared samples are discussed from the results of the FTIR analysis. The formation of nanosphere of diameter of the order of 10 nm, has been confirmed by FESEM. The optical band gap was found by using UV-Visible spectrum. From photoluminescence spectra, a strong emission was observed. The obtained results suggest that this method provides a simple, efficient and versatile technique for preparing TiO₂ nanoparticles and it has the potential to be applied to other systems for photocatalytic activity.

Keywords: TiO2 nanoparticles, chemical precipitation route, phase transition, Fourier Transform Infra-Red spectroscopy (FTIR), micro-Raman spectroscopy, UV-Visible absorption spectroscopy (UV-Vis), Photoluminescence Spectroscopy (PL) and Field Effect Scanning electron microscopy (FESEM)

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Authors: Chakrapani Pullagummi, Arun Jyothi Bheemagani, B. Chandra Sekhar Singh, Prem Kumar, A. Roja Rani

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Diabetes mellitus describes a metabolic disorder of multiple aetiology characterized by chronic hyperglycaemia with disturbances of carbohydrate, fat and protein metabolism resulting from defects in insulin secretion and its action. The objective of present study was alloxan induced diabetes in S.D (Sprague Dawley) rats, treated with leaf extract of Andrographis paniculata and bark extract of Erythrina indica. Plant extract treated rats were analyzed biochemically and molecularly. on normal and diabetic rats. The changes in MDA (lipid peroxidation) and glucose (by GOD method) levels in blood of both normal and diabetic rat were analyzed. Diabetes induced rats were treated with methanolic extracts of Andrographis paniculata leaf and Erythrina indica bark which are of medicinal importance. Later after inducing diabetes the rats were treated with medicinal plant extracts, Andrographis paniculata leaf and Erythrina indica bark which are well known for their anti diabetic and antioxidative property in order to control the glucose and MDA levels. The blood plasma of diabetic and normal rats was analyzed for the levels of MDA (lipid peroxidation) and glucose levels. Results of this study suggested that the Andrographis paniculata leaf and Erythrina indica can be used as a potential natural antidiabetic agent for treating and postponing the appearance of complications that arise due to Diabetes. Molecular study deals with the analysis of binding mechanism of 2 selected natural compounds from Andrographis and Erythrina extracts against the novel target for type T2D namely PPAR-γ compared with Rosiglitazone (standard compound). The results revealed that most of the selected herbal lead compounds were effective targets against the receptors. These compounds showed favorable interactions with the amino acid residues thereby substantiating their proven efficacy as anti-diabetic compounds.

Keywords: andrographis paniculata, erythrina indica, alloxan, lipid peroxidation, blood glucose level, PPAR-γ

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Authors: Mohd Zaki Hamzah, Roslan Rani, Ahmad Bazli Razali, Satiful Bahri Mamat, Abdul Hadi Ripin, Mohd Harun Esa

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Ever since 1901, forest management and silviculture practices in Malaysia have been frequently reviewed and updated to take into account changes in forest conditions, markets, timber demand/supply and technical advances that can be achieved in industrial processes, logging and forest harvesting, and currently, the forest management system practiced in Peninsular Malaysia is the Selective Management System (SMS) which was introduced in 1978. This system requires the selection of management regime (felling) based on Pre-Felling Forest Inventory (Pre-F) data to ensure economical harvesting and also ensuring adequate standing stands for subsequent rounds of felling, while maintaining ecological balance and environmental quality. SMS regulates forest harvesting through area and volume controls, with the cutting cycle 30 years. Most of the forest management units (FMU) (in Peninsular Malaysia) implementing SMS have been certified by Forest Stewardship Council (FSC) and/or Program for Endorsement of Forest Certification (PEFC), and one such FMU belongs to Kumpulan Pengurusan Kayu Kayan Terengganu (KPKKT). KPKKT, a timber management subsidiary of Golden Pharos Berhad (GPB), adopts the SMS to manage its 108,900 ha of timber concessionary areas in its role as logs’ supplier for the consumption of three subsidiaries of GPB. KPKKT is also responsible for the sustainable development and management of its concession in accordance with the Sustainable Forest Management (SFM) standards to ensure that it addresses the loss of forest cover and forest degradation, forest-based economic, social and environmental benefits, and ecologically protecting forests while mobilising financial resources for the implementation of sustainable forest management planning, harvesting, monitoring and the marketing of products. This paper will detail out the management and harvesting guidelines imposed by the controlling government agency, and harvesting processes taken by KPKKT to comply with guidelines and eventually supplying timber to the relevant subsidiaries (downstream mills under GPB).

Keywords: sustainable forest management, silviculture, reduce impact logging, forest certification

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Authors: Kumar Gular, Viswanathan S., Mastour Saeed Alshahrani, Ravi Shankar Reddy, Jaya Shanker Tedla, Snehil Dixit, Ajay Prasad Gautam, Venkata Nagaraj Kakaraparthi, Devika Rani Sangadala

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Background: Despite trunk control’s significant contribution to improving various functional activity components, the independent effect of trunk performance on quality of life is yet to be estimated in stroke survivors. Ascertaining the correlation between trunk control and self-reported quality of life while evaluating the effect of demographic and clinical characteristics on their relationship will guide concerned healthcare professionals in designing ideal rehabilitation protocols during the late sub-acute stroke stage of recovery. The aims of the present research were to (1) investigate the associations of trunk performance with self-rated quality of life and (2) evaluate if age, body mass index (BMI), and clinical characteristics mediate the relationship between trunk motor performance and perceived quality of life in the sub-acute stroke population. Methods: Trunk motor functions and quality of life among the late sub-acute stroke population aged 57.53 ± 6.42 years were evaluated through the trunk Impairment Scale (TIS) and Stroke specific quality of life (SSQOL) questionnaire, respectively. Pearson correlation coefficients and mediation analysis were performed to elucidate the relationship of trunk motor function with quality of life and determine the mediation impact of demographic and clinical characteristics on their association, respectively. Results: The current study observed significant correlations between trunk motor functions (TIS) and quality of life (SSQOL) with r=0.68 (p<0.001). Age, BMI, and type of stroke were detected as potential mediating factors in the association between trunk performance and quality of life. Conclusion: Validated associations between trunk motor functions and perceived quality of life among the late sub-acute stroke population emphasize the importance of comprehensive evaluation of trunk control. Rehabilitation specialists should focus on appropriate strategies to enhance trunk performance anticipating the potential effects of age, BMI, and type of stroke to improve health-related quality of life in stroke survivors.

Keywords: sub-acute stroke, quality of life, functional independence, trunk control

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Authors: Shaik Asha Begum, S. Joshna Rani, Shaik Abdul Rahaman

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INTRODUCTION: Breast cancer is a disease that creates in breast cells. "KAP" study estimates the Knowledge, Attitude, and Practices of a local area. More than 1.5 million ladies (25% of all ladies with malignancy) are determined to have bosom disease consistently all through the world. Understanding the degrees of Knowledge, Attitude and Practice will empower a more effective cycle of mindfulness creation as it will permit the program to be custom-made all the more properly to the necessities of the local area. OBJECTIVES: The objective of this study is to assess the knowledge on signs and symptoms, risk factors, provide awareness on the practicing of the early detection techniques of breast cancer and provide knowledge on the overall breast cancer including preventive techniques. METHODOLOGY: This is an expressive cross-sectional investigation. This investigation of KAP was done in the Nirmala Educational Institutions from January to April 2021. A total of 300 participants are included from women students in pharmacy graduates & lecturers, and also from graduates other than the pharmacy. The examiners are taken from the BCAM (Breast Cancer Awareness Measure), tool compartment (Version 2). RESULT: According to the findings of the study, the majority of the participants were not well informed about breast cancer. A lump in the breast was the most commonly mentioned sign of breast cancer, followed by pain in the breast or nipple. The percentage of knowledge related to the breast cancer risk factors was also very less. The correct answers for breast cancer risk factors were radiation exposure (58.20 percent), a positive family history (47.6 percent), obesity (46.9 percent), a lack of physical activity (43.6 percent), and smoking (43.2 percent). Breast cancer screening, on the other hand, was uncommon (only 30 and 11.3 percent practiced clinical breast examination and mammography respectively). CONCLUSION: In this study, the knowledge on the signs and symptoms, risk factors of breast cancer - pharmacy graduates have more knowledge than the non-pharmacy graduates but in the preventive techniques and early detective tools of breast cancer -had poor knowledge in the pharmacy and non-pharmacy graduate. After the awareness program, pharmacy and non-pharmacy graduates got supportive knowledge on the preventive techniques and also practiced the early detective techniques of breast cancer.

Keywords: breast cancer, mammography, KAP study, early detection

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Authors: Sanjay Kumar Srivastava, Anu Rani Sharma, Kamna Sachdeva

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Every year, fog formation over the Indo-Gangetic Plains (IGPs) of Indian region during the winter months of December and January is believed to create numerous hazards, inconvenience, and economic loss to the inhabitants of this densely populated region of Indian subcontinent. The aim of the paper is to analyze the spatial and temporal variability of winter fog over IGPs. Long term ground observations of visibility and other meteorological parameters (1971-2010) have been analyzed to understand the formation of fog phenomena and its relevance during the peak winter months of January and December over IGP of India. In order to examine the temporal variability, time series and trend analysis were carried out by using the Mann-Kendall Statistical test. Trend analysis performed by using the Mann-Kendall test, accepts the alternate hypothesis with 95% confidence level indicating that there exists a trend. Kendall tau’s statistics showed that there exists a positive correlation between time series and fog frequency. Further, the Theil and Sen’s median slope estimate showed that the magnitude of trend is positive. Magnitude is higher during January compared to December for the entire IGP except in December when it is high over the western IGP. Decade wise time series analysis revealed that there has been continuous increase in fog days. The net overall increase of 99 % was observed over IGP in last four decades. Diurnal variability and average daily persistence were computed by using descriptive statistical techniques. Geo-statistical analysis of fog was carried out to understand the spatial variability of fog. Geo-statistical analysis of fog revealed that IGP is a high fog prone zone with fog occurrence frequency of more than 66% days during the study period. Diurnal variability indicates the peak occurrence of fog is between 06:00 and 10:00 local time and average daily fog persistence extends to 5 to 7 hours during the peak winter season. The results would offer a new perspective to take proactive measures in reducing the irreparable damage that could be caused due to changing trends of fog.

Keywords: fog, climatology, Mann-Kendall test, trend analysis, spatial variability, temporal variability, visibility

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Authors: Anu Chandran, Reena Esther Rani

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Background of the Study: Dotted with multiple Unique Destination Prepositions (UDPs), Tamil Nadu is an established tourism brand as regards leisure, MICE, culture, and ecological flavors. Albeit, the enchanting destination possesses distinctive attributes and resources yet to be tapped for better competitive advantage. Being a destination that allures an incredible variety of migratory birds, Tamil Nadu is deemed to be an ornithologist’s paradise. This study primarily explores the prospects of developing Kaliveli, recognized as a bird sanctuary in the Tindivanam forest division of the Villupuram district in the State. Kaliveli is an ideal nesting site for migratory birds and is currently apt for a prospective analysis of regenerative tourism. Objectives of the study: This research lays an accent on avian tourism as part and parcel of sustainable tourism ventures. The impacts of projects like the Ornithological Conservation Centre on tourists have been gauged in the present paper. It maps the futuristic proactive propositions linked to regenerative tourism on the site. How far technological innovations can do a world of good in Kaliveli through Artificial Intelligence, Smart Tourism, and similar latest coinages to entice real eco-tourists, have been conceptualized. The experiential dimensions of resource stewardship as regards facilitating tourists’ relish the offerings in a sustainable manner is at the crux of this work. Methodology: Modeled as a case study, this work tries to deliberate on the impact of existing projects attributed to avian fauna in Kalveli. Conducted in the qualitative research design mode, the case study method was adopted for the processing and presentation of study results drawn by applying thematic content analysis based on the data collected from the field. Result and discussion: One of the key findings relates to the kind of nature trails that can be a regenerative dynamic for eco-friendly tourism in Kaliveli. Field visits have been conducted to assess the niche tourism aspects which could be incorporated with the regenerative tourism model to be framed as part of the study.

Keywords: regenerative tourism, Kaliveli bird sanctuary, sustainable development, resource Stewardship, Ornithology, Avian Fauna

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Authors: Rooma Rani

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The education is really important for the improvement of physical and mental well-being of the school students. It is used to express inner potential, acquire knowledge, develop skills, shape habits, attitudes, values, belief, etc. along with providing strengths and resilience to people to changing situations and allowing them to develop all those capacities which will enable individual to control surrounding environment. Education has a significant effect on the behavior of individuals which helps us in the new situations of everyday life. Educating the child is directing the child’s capacities, attitudes interest, urges, and needs into the most desirable channels. We are the part of 21st century and now a day emotional intelligence is considered more important than intelligence in the success of a person. Success depends on several intelligences and on the control of emotions too. Emotional Intelligence, like general intelligence is the product of one’s heredity and its interaction with his environmental forces. There are certain methods evolved in modern researches. Keeping in view the nature and purpose of the study, the descriptive survey method is preferred. This method is one of the important methods in education research because it describes the current position of the phenomenon under study. The term descriptive survey is generally used for the type of research which proposes to condition of practices of the present time. In the present study, a systematically random sampling method was used to select a representative sample. 50 students were selected from 2 schools. Out of 50 students, 25 were boys and 25 were girls. In the study, a) it has been found a significant difference in the level of adjustment between male and female students; b) it has been found a non-significant difference in the level of emotional intelligence between male and female students; c) it has been found a non-significant relationship between adjustment and emotional intelligence among male students; d) it has been found a significant relationship between adjustment and emotional intelligence among male students. The results of the study indicated that amongst the students those who possess high scores on emotional intelligence tests are high in level of adjustment. Measures should be adopted to improve and sustain the emotional intelligence level of students throughout their studies. Adolescent students are prone to many problems like physical, social and psychological. They need a congenial home atmosphere so that they grow into full-fledged citizens of our country. After understanding these, it helps in the development of personality which leads to a better learning situation and better thinking capacities, in turn, enhances adjustment and achievement along with a better perception of self.

Keywords: adjustment, education, emotional intelligence, students

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Authors: S. Agarwal, A. Rani

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Catechol is a natural polyphenolic compound that widely exists in higher plants such as teas, vegetables, fruits, tobaccos, and some traditional Chinese medicines. The fly ash-based zeolites are capable of absorbing a wide range of pollutants. But the process of zeolite synthesis is time-consuming and requires technical setups by the industries. The marketed costs of zeolites are quite high restricting its use by small-scale industries for the removal of phenolic compounds. The present research proposes a simple method of alkaline treatment of FA to produce an effective adsorbent for catechol removal from wastewater. The experimental parameter such as pH, temperature, initial concentration and adsorbent dose on the removal of catechol were studied in batch reactor. For this purpose the adsorbent materials were mixed with aqueous solutions containing catechol ranging in 50 – 200 mg/L initial concentrations and then shaken continuously in a thermostatic Orbital Incubator Shaker at 30 ± 0.1 °C for 24 h. The samples were withdrawn from the shaker at predetermined time interval and separated by centrifugation (Centrifuge machine MBL-20) at 2000 rpm for 4 min. to yield a clear supernatant for analysis of the equilibrium concentrations of the solutes. The concentrations were measured with Double Beam UV/Visible spectrophotometer (model Spectrscan UV 2600/02) at the wavelength of 275 nm for catechol. In the present study, the use of low-cost adsorbent (BTFA) derived from coal fly ash (FA), has been investigated as a substitute of expensive methods for the sequestration of catechol. The FA and BTFA adsorbents were well characterized by XRF, FE-SEM with EDX, FTIR, and surface area and porosity measurement which proves the chemical constituents, functional groups and morphology of the adsorbents. The catechol adsorption capacities of synthesized BTFA and native material were determined. The adsorption was slightly increased with an increase in pH value. The monolayer adsorption capacities of FA and BTFA for catechol were 100 mg g⁻¹ and 333.33 mg g⁻¹ respectively, and maximum adsorption occurs within 60 minutes for both adsorbents used in this test. The equilibrium data are fitted by Freundlich isotherm found on the basis of error analysis (RMSE, SSE, and χ²). Adsorption was found to be spontaneous and exothermic on the basis of thermodynamic parameters (ΔG°, ΔS°, and ΔH°). Pseudo-second-order kinetic model better fitted the data for both FA and BTFA. BTFA showed large adsorptive characteristics, high separation selectivity, and excellent recyclability than FA. These findings indicate that BTFA could be employed as an effective and inexpensive adsorbent for the removal of catechol from wastewater.

Keywords: catechol, fly ash, isotherms, kinetics, thermodynamic parameters

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Authors: Monika Rani, Claudio Marchesi, Stefania Federici, Laura E. Depero

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Degradation of the aquatic environment by plastic litter, especially microplastics (MPs), i.e., any water-insoluble solid plastic particle with the longest dimension in the range 1µm and 1000 µm (=1 mm) size, is an unfortunate indication of the advancement of the Anthropocene age on Earth. Microplastics formed due to natural weathering processes are termed as secondary microplastics, while when these are synthesized in industries, they are called primary microplastics. Their presence from the highest peaks to the deepest points in oceans explored and their resistance to biological and chemical decay has adversely affected the environment, especially marine life. Even though the presence of MPs in the marine environment is well-reported, a legitimate and authentic analytical technique to sample, analyze, and quantify the MPs is still under progress and testing stages. Among the characterization techniques, vibrational spectroscopic techniques are largely adopted in the field of polymers. And the ongoing miniaturization of these methods is on the way to revolutionize the plastic recycling industry. In this scenario, the capability and the feasibility of a miniaturized near-infrared (MicroNIR) spectroscopy combined with chemometrics tools for qualitative and quantitative analysis of urban plastic waste collected from a recycling plant and microplastic mixture fragmented in the lab were investigated. Based on the Resin Identification Code, 250 plastic samples were used for macroplastic analysis and to set up a library of polymers. Subsequently, MicroNIR spectra were analysed through the application of multivariate modelling. Principal Components Analysis (PCA) was used as an unsupervised tool to find trends within the data. After the exploratory PCA analysis, a supervised classification tool was applied in order to distinguish the different plastic classes, and a database containing the NIR spectra of polymers was made. For the microplastic analysis, the three most abundant polymers in the plastic litter, PE, PP, PS, were mechanically fragmented in the laboratory to micron size. The distinctive arrangement of blends of these three microplastics was prepared in line with a designed ternary composition plot. After the PCA exploratory analysis, a quantitative model Partial Least Squares Regression (PLSR) allowed to predict the percentage of microplastics in the mixtures. With a complete dataset of 63 compositions, PLS was calibrated with 42 data-points. The model was used to predict the composition of 21 unknown mixtures of the test set. The advantage of the consolidated NIR Chemometric approach lies in the quick evaluation of whether the sample is macro or micro, contaminated, coloured or not, and with no sample pre-treatment. The technique can be utilized with bigger example volumes and even considers an on-site evaluation and in this manner satisfies the need for a high-throughput strategy.

Keywords: chemometrics, microNIR, microplastics, urban plastic waste

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Authors: Anupalli Roja Rani, Pavithra Dasari

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Background: Among several, breast cancer has emerged as the most common female cancer in developing countries. It is the most common cause of cancer-related deaths worldwide among women. It is a molecularly and clinically heterogeneous disease. Moreover, it is a hormone–dependent tumor in which estrogens can regulate the growth of breast cells by binding with estrogen receptors (ERs). Moreover, the use of natural products in cancer therapeutics is due to their properties of biocompatibility and less toxicity. Plants are the vast reservoirs for various bioactive compounds. Coleus barbatus (Lamiaceae) contains anticancer properties against several cancer cell lines. Method: In the present study, an attempt is being made to enrich the knowledge of the anticancer activity of pure compounds extracted from Coleus barbatus (Andrew). On human breast cancer cell lines MCF-7. Here in, we are assessing the antiproliferative activity of Coleus barbatus (Andrew) plant extracts against MCF 7 and also evaluating their toxicity in normal human mammary cell lines such as Human Mammary Epithelial Cells (HMEC). The active fraction of plant extract was further purified with the help of Flash chromatography, Medium Pressure Liquid Chromatography (MPLC) and preparative High-Performance Liquid Chromatography (HPLC). The structure of pure compounds will be elucidated by using modern spectroscopic methods like Nuclear magnetic resonance (NMR), Electrospray Ionisation Mass Spectrometry (ESI-MS) methods. Later, the growth inhibition morphological assessment of cancer cells and cell cycle analysis of purified compounds were assessed using FACS. The growth and progression of signaling molecules HER2, GRP78 was studied by secretion assay using ELISA and expression analysis by flow cytometry. Result: Cytotoxic effect against MCF-7 with IC50 values were derived from dose response curves, using six concentrations of twofold serially diluted samples, by SOFTMax Pro software (Molecular device) and respectively Ellipticine and 0.5% DMSO were used as a positive and negative control. Conclusion: The present study shows the significance of various bioactive compounds extracted from Coleus barbatus (Andrew) root material. It acts as an anti-proliferative and shows cytotoxic effects on human breast cancer cell lines MCF7. The plant extracts play an important role pharmacologically. The whole plant has been used in traditional medicine for decades and the studies done have authenticated the practice. Earlier, as described, the plant has been used in the ayurveda and homeopathy medicine. However, more clinical and pathological studies must be conducted to investigate the unexploited potential of the plant. These studies will be very useful for drug designing in the future.

Keywords: coleus barbatus, HPLC, MPLC, NMR, MCF7, flash chromatograph, ESI-MS, FACS, ELISA.

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Authors: Ved Vrat Verma, Rani Gupta, Manisha Goel

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γ-glutamyltranspeptidase (GGT: EC 2.3.2.2) is an N-terminal nucleophile hydrolase conserved in all three domains of life. GGT plays a key role in glutathione metabolism where it catalyzes the breakage of the γ-glutamyl bonds and transfer of γ-glutamyl group to water (hydrolytic activity) or amino acids or short peptides (transpeptidase activity). GGTs from bacteria, archaea, and eukaryotes (human, rat and mouse) are homologous proteins sharing >50% sequence similarity and conserved four layered αββα sandwich like three dimensional structural fold. These proteins though similar in their structure to each other, are quite diverse in their enzyme activity: some GGTs are better at hydrolysis reactions but poor in transpeptidase activity, whereas many others may show opposite behaviour. GGT is known to be involved in various diseases like asthma, parkinson, arthritis, and gastric cancer. Its inhibition prior to chemotherapy treatments has been shown to sensitize tumours to the treatment. Microbial GGT is known to be a virulence factor too, important for the colonization of bacteria in host. However, all known inhibitors (mimics of its native substrate, glutamate) are highly toxic because they interfere with other enzyme pathways. However, a few successful efforts have been reported previously in designing species specific inhibitors. We aim to leverage the diversity seen in GGT family (pathogen vs. eukaryotes) for designing specific inhibitors. Thus, in the present study, we have used DIVERGE software to identify sites in GGT proteins, which are crucial for the functional and structural divergence of these proteins. Since, type II divergence sites vary in clade specific manner, so type II divergent sites were our focus of interest throughout the study. Type II divergent sites were identified for pathogen vs. eukaryotes clusters and sites were marked on clade specific representative structures HpGGT (2QM6) and HmGGT (4ZCG) of pathogen and eukaryotes clade respectively. The crucial divergent sites within 15 A radii of the binding cavity were highlighted, and in-silico mutations were performed on these sites to delineate the role of these sites on the mechanism of catalysis and protein folding. Further, the amino acid network (AAN) analysis was also performed by Cytoscape to delineate assortative mixing for cavity divergent sites which could strengthen our hypothesis. Additionally, molecular dynamics simulations were performed for wild complexes and mutant complexes close to physiological conditions (pH 7.0, 0.1 M ionic strength and 1 atm pressure) and the role of putative divergence sites and structural integrities of the homologous proteins have been analysed. The dynamics data were scrutinized in terms of RMSD, RMSF, non-native H-bonds and salt bridges. The RMSD, RMSF fluctuations of proteins complexes are compared, and the changes at protein ligand binding sites were highlighted. The outcomes of our study highlighted some crucial divergent sites which could be used for novel inhibitors designing in a species-specific manner. Since, for drug development, it is challenging to design novel drug by targeting similar protein which exists in eukaryotes, so this study could set up an initial platform to overcome this challenge and help to deduce the more effective targets for novel drug discovery.

Keywords: γ-glutamyltranspeptidase, divergence, species-specific, drug design

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Authors: Shikha Rani, Neeta Sehgal, Vipin Kumar Verma, Om Prakash

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Introduction: Fish is an important protein source for humans and has great economic value. Fish cultures are affected due to various anthropogenic activities that lead to bacterial and viral infections. Aeromonas hydrophila is a fish pathogenic bacterium that causes several aquaculture outbreaks throughout the world and leads to huge mortalities. In this study, plants of no commercial value were used to investigate their immunostimulatory, antioxidant, anti-inflammatory, anti-bacterial, and disease resistance potential in fish against Aeromonas hydrophila, through fish feed fortification. Methods: The plant was dried at room temperature in the shade, dissolved in methanol, and analysed for biological compounds through GC-MS/MS. DPPH, FRAP, Phenolic, and flavonoids were estimated following standardized protocols. In silico molecular docking was also performed to validate its broad-spectrum activities based on binding affinity with specific proteins. Fish were divided into four groups (n=6; total 30 in a group): Group 1, non-challenged fish (fed on a non-supplemented diet); Group 2, fish challenged with bacteria (fed on a non-supplemented diet); Group 3 and 4, fish challenged with bacteria (A. hydrophila) and fed on plant supplemented feed at 2.5% and 5%. Blood was collected from the fish on 0, 7th, 14th, 21st, and 28th days. Serum was separated for glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase assay (ALP), lysozyme activity assay, superoxide dismutase assay (SOD), lipid peroxidation assay (LPO) and molecular parameters (including cytokine levels) were estimated through ELISA. The phagocytic activity of macrophages from the spleen and head kidney, along with quantitative analysis of immune-related genes, were analysed in different tissue samples. The digestive enzymes (Pepsin, Trypsin, and Chymotrypsin) were also measured to evaluate the effect of plant-supplemented feed on freshwater fish. Results and Discussion: GC-MS/MS analysis of a methanolic extract of plant validated the presence of key compounds having antioxidant, anti-inflammatory, anti-bacterial, anti-inflammatory, and immunomodulatory activities along with disease resistance properties. From biochemical investigations like ABTS, DPPH, and FRAP, the amount of total flavonoids, phenols, and promising binding affinities towards different proteins in molecular docking analysis helped us to realize the potential of this plant that can be used for investigation in the supplemented feed of fish. Measurement liver function tests, ALPs, oxidation-antioxidant enzyme concentrations, and immunoglobulin concentrations in the experimental groups (3 and 4) showed significant improvement as compared to the positive control group. The histopathological evaluation of the liver, spleen, and head kidney supports the biochemical findings. The isolated macrophages from the group fed on supplemented feed showed a higher percentage of phagocytosis and a phagocytic index, indicating an enhanced cell-mediated immune response. Significant improvements in digestive enzymes were also observed in fish fed on supplemented feed, even after weekly challenges with bacteria. Hence, the plant-fortified feed can be recommended as a regular feed to enhance fish immunity and disease resistance against the Aeromonas hydrophila infection after confirmation from the field trial.

Keywords: immunostimulation, antipathogen, plant fortified feed, macrophages, GC-MS/MS, in silico molecular docking

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