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Patil, S. A.

Publications and source records attributed to Patil, S. A..

10 recordsLinked to original sources

Chemo-Profiling by UPLC-QTOF-MS, GC-MS/MS analysis and In Vitro Bioactivity Assessment of Desmodium gangeticum DC.

Desmodium gangeticum DC., commonly known as Shalparni, and a member of the Fabaceae family, is a small shrub native to tropical and subtropical regions. The active principles of this plant are primarily concentrated in its roots, which are traditionally used in various medicinal formulations. This study aimed to identify alternative sources of raw material by evaluating the biochemical analysis and bioactivity profiling of both fresh and dried plant parts--roots, leaves, and stems. Extracts were prepared using methanol, ethanol, and distilled water and subjected to phytochemical, antioxidant, anti-inflammatory (HRBC membrane stabilization), and antibacterial assays. Additionally, chemical profiling was performed using FTIR, GC-MS/MS, and UPLC-Q-TOF-MS. This is the first report on D. gangeticum as one of the Dashmoola plants for its biochemical analysis, bioactivity, and chemoprofiling of the entire plant. Results indicated that the GC-MS/MS analysis identified 25 volatile compounds in leaves, whereas the roots contained 21. These compounds included fatty acids, ketones, hydrocarbons, alcohols, and esters. UPLC-Q-TOF-MS analysis revealed a diverse array of bioactive compounds: 287 in aqueous leaf extracts and 192 in aqueous root extracts, many of which were identified as bioactive agents. Notably, the leaf extracts exhibited a higher number of bioactive compounds compared to the roots, highlighting the plants diverse bioactive compounds in different tissues. The study demonstrates that both the leaves and roots of D. gangeticum contain significant therapeutic compounds. Therefore, the leaves could serve as a sustainable alternative to roots as raw material for medicinal preparations, contributing to the conservation of this plant in its natural habitat.

plant biology↗

Exploring variations in lipids among drug-resistant and sensitive Mycobacterium tuberculosis by Thin layer chromatography and mass spectrometry

Mycobacterium tuberculosis (M.tb) lipids are important in the host-pathogen interplay, variation in the lipids organization of cell wall can act as an adaptive response. Specific cell wall structures can possibly result in suboptimal intracellular concentrations of anti-TB drugs, which favors the acquisition of drug resistance. Therefore, lipids from M.tb (drug resistant and sensitive) were analyzed by 2D-thin layer chromatography and mass spectrometry. GraphPad Prism was used to perform Mann Whitney-U test to determine the statistical significance. Difference observed for total lipid content among different resistant isolates was insignificant. However, increase in phospholipids was identified in multi-drug resistant (MDR) isolate compared to sensitive isolate. Isoniazid, streptomycin-isoniazid, and isoniazid-ethambutol resistant isolates showed increased alpha-mycolic acids. MDR isolate showed a marginal decrease in alpha- and keto-form. Mycolipenic acid was seen only in sensitive isolate, and mycosanoic acids were observed in all the resistant isolates. Among the resistant isolates, there was an insignificant increase in the total phthiocerol dimycocerosates and sulfolipids. Drug resistance was associated with compositional imbalance of lipids. However, investigations to determine whether the changes notices are induced by the drugs is to be explored, which could give an insight into the drug resistant organisms pathogenesis.

microbiology↗

Insights into the genomic features, lifestyle and therapeutic potential of B1 sub-cluster mycobacteriophages

BackgroundA large number (about 1200) of mycobacteriophages (phages) have been isolated on Mycobacterium smegmatis mc2155. Their genome analysis shows high sequence diversity; therefore, based on nucleotide sequence similarity and genomic architecture, the related phages have been grouped in clusters and sub-clusters. However, a deeper study of mycobacteriophages has been conducted only for a few clusters. This study explores the traits of phages belonging to the B1 sub-cluster. We have attempted to functionally annotate and experimentally characterize B1 phages to get an insight into their biology and explore their therapeutic potential. MethodsAnalysis of B1 sub-cluster phage genomes to understand their key characteristics & lifestyle and to determine the putative function of hypothetical proteins (HPs), we developed a framework with a specific set of computational tools available online. For the experimental characterization, mycobacteriophages were isolated from environmental samples and were examined for their morphology, lysogeny status, effect on biofilm and activity against drug-resistant M. smegmatis. The B1 sub-cluster phages were identified by PCR using the specific primers. ResultsWe have predicted the function of about 55% of the 77 representative proteins in B1 phages, which were previously deemed hypothetical. We studied ten B1 phages (Phages 1-10) which included their morphological characteristics, lysogeny status and antibiofilm activity. TEM analysis, showing an average head & tail size of 65 nm and 202.12 nm, respectively. The turbid morphology of several plaques suggested these phages to be temperate. To verify, we tested their potential to lysogenize M. smegmatis and later found the spontaneous release from the putative lysogens. Interestingly, a putative RepA-like protein was identified in B1 phage genomes, indicating a possibility of extrachromosomal replication of prophages. Further, the impact of Phages 1-10 on M. smegmatis biofilm was found to be potent; the highest inhibitory and disruptive effect of phages (at a fixed titre of 108 pfu/ml) was 64% and 46%, respectively. Also, all ten phages could kill 4XR1 (the isoniazid-resistant M. smegmatis strain). ConclusionWe believe this combination of experimental analysis and exploration of genomic features of mycobacteriophages belonging to a sub-cluster can provide deeper insights into mycobacteriophage biology and also help in understanding their therapeutic potential.

microbiology↗

The bi-directional extracellular electron transfer process aids iron cycling by Geoalkalibacter halelectricus in a highly saline-alkaline condition

Bi-directional extracellular electron transfer (EET) is crucial to upholding microbial metabolism with insoluble electron acceptors or donors in anoxic environments. Investigating bi-directional EET-capable microorganisms is desired to understand the cell-cell and microbe-mineral interactions and their role in mineral cycling besides leveraging their energy generation and conversion, biosensing, and bio-battery applications. Here, we report on iron cycling by haloalkaliphilic Geoalkalibacter halelectricus via bi-directional EET under haloalkaline conditions. It efficiently reduces Fe3+-oxide (Fe2O3) to Fe0 at a 2.29{+/-}0.07 mM/day rate linked to acetate oxidation via outward EET and oxidizes Fe0 to Fe3+ with a 0.038{+/-}0.002 mM/day rate via inward EET to reduce fumarate. Bioelectrochemical cultivation confirmed its outward and inward EET capabilities. It produced 895{+/-}23 A/cm2 current by linking acetate oxidation to anode reduction via outward EET and reduced fumarate by drawing electrons from the cathode (-2.5{+/-}0.3 A/cm2) via inward EET. The cyclic voltammograms of G. halelectricus biofilms revealed redox moieties with different formal potentials, suggesting the involvement of different membrane components in bi-directional EET. The cyclic voltammetry and GC-MS analysis of the cell-free spent medium revealed the lack of soluble redox mediators, suggesting direct electron transfer by G. halelecctricus in achieving bi-directional EET. By reporting on the first haloalkaliphilic bacterium capable of oxidizing and reducing insoluble Fe0 and Fe3+-oxide, respectively, this study advances the limited understanding of the metabolic capabilities of extremophiles to respire on insoluble electron acceptors or donors via bi-directional EET and invokes the possible role of G. halelectricus in iron cycling in barely studied haloalkaline environments. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/536630v1_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@1c8080aorg.highwire.dtl.DTLVardef@187d9d9org.highwire.dtl.DTLVardef@1ef6066org.highwire.dtl.DTLVardef@a2081b_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

PHYTOREMEDIATION OF REACTIVE TURQUOISE BLUE H5G USING IN VITRO CULTURES OF SOLANUM VIRGINIANUM (L.)

Solanum virginianum (L.) belonging to family Solanaceae selected for decolourisation of Reactive Turquoise Blue H5G dye. In vitro grown cultures of S. virginianum were able to remove more than 50% dye concentration up to 110mg/l. comparative analysis of biochemical and antioxidant study showed more activity in treated plants as compared to untreated plants. The phytotoxicity study demonstrated the non-toxic nature of degraded metabolites. Use of such non-edible yet medicinal plant for phytoremediation is discussed.

plant biology↗

Biosynthesis of Silver nanoparticles using Drimia indica and exploring its antibacterial profiling

Biological synthesis reflects as an eco-friendly, nontoxic and easy method of nanoparticle preparation. Present investigation deals with biosynthesis of silver nanoparticles using Drimia indica leaf extract. Initially the synthesized silver nanoparticles were characterized and confirmed by UV-Vis spectroscopy, X-ray diffraction (XRD) and SEM. The synthesized nanoparticles when used for determination of antibacterial activity, by Microtitre Broth Dilution method exhibited remarkable activity against Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli.

plant biology↗

REGENERATION VIA SOMATIC EMBRYOGENESIS FROM SEED EXPLANT OF MEDICINAL PLANT SOLANUM VIRGINIANUM (L.)

Solanum virginianum (L.) belonging to family Solanaceae is an important medicinal plant, used in traditional ayurvedic medicines. It is one of the ten constituents of "Dashmoola". Present work deals with the development of efficient procedure for inducing somatic embryogenesis followed by regeneration of Solanum virginianum (L.). Initially, seeds of S. virginianum were used as explant and inoculated on [1/2] MS basal medium. The stem explant from in vitro germinated seeds was used for callus induction. MS medium supplemented with BAP (1mg/L) was used for callus induction. Callus proliferation started when initiated callus was subcultured on different concentrations and combination of BAP, IAA, IBA, 2,4-D and NAA. Where highest embryogenesis took place in BAP (2mg/l)+IAA(5mg/l) combination. The somatic embryos were transferred onto MS basal medium for maturation. These embryos matured and were converted into complete plantlets with well developed shoot and root system.

plant biology↗

Cytotoxic effect of Rotheca serrata on cancer cell lines MCF-7 and Neuroblastoma SH-SY5Y

Rotheca serrata (Lamiaceae), a highly medicinal plant is used as an antidote for snakebite and the plant possesses medicinal properties like hepatoprotective, antitussive, antioxidant, anticancer, neuro-protective, used in rheumatoid arthritis and is also a -glucoside inhibitor. This work aimed to study the anticancerous effect of Rotheca serrata (root and leaf) on cancer cell lines MCF-7 (breast cancer cell line) and Neuroblastoma SH-SY5Y. The results indicated that the Methanolic extract of Rotheca serrata (root and leaf) showed high anticancer activity. Different concentrations of plant extracts (25, 50, 100, 200, 400 g/ml) were used to study the anticancerous activity, amongst which the significant results were obtained for 400 g/ml concentration (both root & leaf). Effective anticancer activity against MCF - 7 breast cancer cells was shown in methanoilc extracts and were expressed as IC 50 values; in root (IC 50 value= 61.8259 {+/-} 7.428 g/ml) and in leaf (IC 50 value = 78.1497 {+/-} 6.316g/ml). The MTT assay in case of neuroblastoma (SH-SY5Y) cell lines revealed that 400g/ml concentration of leaf methanolic extract showed effective inhibition of cancer cells with IC 50 value 37.8462 {+/-} 2.957 g/ml as compared to IC 50 value of root methanolic extract which was 57.0895 {+/-} 2.351 g/ml.

cancer biology↗

Comparative evaluation of plasma protein purification and 2-D gel electrophoresis protocols for analysis of HIV-1 infected human plasma proteins

Purification of proteins from human plasma is a herculean task to perform 2-D gel electrophoresis. Human plasma contains nearly 70% albumin and globulin. The removal of such high abundance high molecular weight proteins is very difficult before performing 2-D gel electrophoresis. It becomes more difficult when we intent to investigate in infectious diseases like HIV/AIDS. We tried to the best of our efforts adopting various organic and non-organic based protocols based on various published papers. After failure of these protocols in results of 2-D gel-electrophoresis Aurum serum mini kit (Bio-Rad, USA) was adopted for plasma protein purification for performing 2-D gel electrophoresis. The low-abundance proteins were better resolved by 10% SDS-PAGE in 2-D gel-electrophoresis. Then,we extended the MALDI-TOF/TOF analysis of low-abundance proteins in human plasma by adopting the Aurum serum mini kit (Bio-Rad, USA) for 2-D gel electrophoresis. Thus, we concluded that, depletion of high abundant proteins like albumin and globulin, the use of the Aurum serum mini kit (Bio-Rad, USA) is the protocol of choice to perform the 2-D gel electrophoresis of HIV-1 infected human plasma.

biochemistry↗

Identification and characterization of novel plasma proteins in drug resistant HIV/AIDS patients by SWATH-MS

Background and ObjectivesAcquired immunodeficiency syndrome is one of the most important diseases caused by human immunodeficiency virus. Understanding its molecular pathogenesis is essential to manage the disease at the population level. In this study, a quantitative analysis of plasma proteins was carried out in drug resistant and drug respondent patients using the SWATH-MS. MethodsSequential window acquisition of all theoretical mass spectra (SWATH-MS) is a prime technique to seek the key plasma proteins involved in virus replication and drug metabolism during therapy. ResultsIn total, 204 proteins were identified and quantified, 57 proteins were differentially expressed, 25 proteins were down regulated and 32 proteins were upregulated in drug resistant patients. Six proteins such as complement C4-A, immunoglobulin heavy variable 1-2, carboxylic ester hydrolase, fibulin-1, immunoglobulin lambda constant 7, secreted phosphoprotein 24 were statistically expressed in drug resistant patients compared to the drug respondent patients. Gene ontology study and protein-protein interaction networks were established in six statistically significant differentially expressed proteins of the drug resistant patients. Interpretation and ConclusionOur findings high lights the novel proteins that were differentially expressed in drug resistant patients. A label-free quantitative proteomics method for depleted human plasma samples by SWATH-MS that can be useful in plasma proteomics research in any biological system.

biochemistry↗