A new, simple, sensitive, selective, rapid, and high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for simultaneous quantification of ...Olmesartan and hydrochlorothiazide in human plasma. Simple liquid-liquid extraction procedure was applied for plasma sample pretreatment using a mixture of diethyl ether and dichloromethane, as an extraction solution. Analytes were separated on UNISOL C18 150*4.6 mm, 5 µm column using methanol, and 2 mM ammonium acetate pH 5.5 (80:20, v/v) as a mobile phase and detected by electrospray ionization in the multiple reaction monitoring (MRM) mode. The mass transition ion pairs were followed in negative ion mode as m/z 445.20 → 148.90 for Olmesartan; m/z 451.40 → 154.30 for Olmesartan D
and m/z 295.80 → 205.10 for hydrochlorothiazide; m/z 298.90 → 206.30 for hydrochlorothiazide
C D
. The method showed excellent linearity (
> 0.99) over the concentration range of 5.002-2,599.934 ng/ml for Olmesartan and from 3.005 to 499.994 ng/ml for hydrochlorothiazide. Precision (% CV) and accuracy (% bias) for Olmesartan were found in the range of 3.07-9.02% and -5.00-0.00%, respectively. Precision (% CV) and accuracy (% bias) for hydrochlorothiazide were found in the range of 3.32-8.21% and 1.99-3.80%, respectively. This as developed novel and high-throughput liquid-liquid extraction bioanalytical method has substantial innovative value with the benefits of cost effectiveness, good extraction efficiency, shorter analysis run time, low organic solvent consumption, and simpler procedure over the previously reported solid-phase extraction method. The application of this method in pharmacokinetic studies was further demonstrated successfully through a bioequivalence study conducted on healthy human subjects, following oral administration of combined formulation of Olmesartan medoxomil and hydrochlorothiazide in fixed-dose tablet.
Phytoplankton are the world's largest oxygen producers found in oceans, seas and large water bodies, which play crucial roles in the marine food chain. Unbalanced biogeochemical features like ...salinity, pH, minerals,
., can retard their growth. With advancements in better hardware, the usage of Artificial Intelligence techniques is rapidly increasing for creating an intelligent decision-making system. Therefore, we attempt to overcome this gap by using supervised regressions on reanalysis data targeting global phytoplankton levels in global waters. The presented experiment proposes the applications of different supervised machine learning regression techniques such as random forest, extra trees, bagging and histogram-based gradient boosting regressor on reanalysis data obtained from the Copernicus Global Ocean Biogeochemistry Hindcast dataset. Results obtained from the experiment have predicted the phytoplankton levels with a coefficient of determination score (R
) of up to 0.96. After further validation with larger datasets, the model can be deployed in a production environment in an attempt to complement
measurement efforts.
Abstract
During survey from 2013 to 2018,
Cynodon dactylon
at Shahjahnapur, Azamagarh,
Oplismenus burmannii
at Shahjahnapur,
Arundo donax
at Deoria and
Ocimum cannum
at Gorakhpur were observed with ...phytoplasma suspected symptoms. The maximum disease incidence (34%) was recorded at Deoria District in case of
A. donax
plant with witches' broom symptom. The nested PCR using universal primer pairs (R16F2n/R16R2) confirmed the phytoplasma association with all the suspected samples. Further BLASTn and phylogenetic analysis of the sequences revealed the association of 16SrXIV-A subgroups phytoplasma with
C. dactylon
,
O. burmannii
and phytoplasma belonging to 16SrI-B subgroups with
A. donax
and
O. cannum
plants in the present study.
Cryptosporidium parvum is the common enteric protozoan pathogen causing cryptosporidiosis in human. Available drugs to treat cryptosporidiosis are ineffective and there is yet no vaccine against C. ...parvum. Therefore, it is of interest to design an improved yet effective drug against C. parvum. Here, we docked benzoxazole derivatives (collected from literature) with inosine 5`- monophosphate dehydrogenase (IMPDH) from Cryptosporidium parvum using the program AutoDock 4.2. The docked protein - inhibitor complex structure was optimized using molecular dynamics simulation for 5 ps with the CHARMM-22 force field using NAMD (NAnoscale Molecular Dynamics program) incorporated in visual molecular dynamics (VMD 1.9.2) and then evaluating the stability of complex structure by calculating RMSD values. NAMD is a parallel, object-oriented molecular dynamics code designed for high-performance simulation of large biomolecular systems. A quantitative structure activity relationship (QSAR) model was built using energy-based descriptors as independent variable and pIC50 value as dependent variable of fifteen known benzoxazole derivatives with C. parvum IMPDH protein, yielding correlation coefficient r2 of 0.7948. The predictive performance of QSAR model was assessed using different cross-validation procedures. Our results suggest that a ligand-receptor binding interaction for inosine 5`-monophosphate dehydrogenase using a QSAR model is promising approach to design more potent inosine 5`-monophosphate dehydrogenase inhibitors prior to their synthesis.
A significant increase in the incidence of fungal infections and drug resistance has been observed in the past decades due to limited availability of broad-spectrum antifungal drugs. Nanomedicines ...have shown significant antimicrobial potential against various drug-resistant microbes. Silver nanoparticles (AgNps) are known for their antimicrobial properties and lower host toxicity; however, for clinical applications, evaluation of their impact at cellular and molecular levels is essential. The present study aims to understand the cellular and molecular mechanisms of AgNp-induced toxicity in a common fungal pathogen,
.
AgNps were synthesized by chemical reduction method and characterized using UV-visible spectroscopy, X-ray powder diffraction, transmission electron microscopy, scanning electron microscopy-energy dispersive X-ray spectroscopy, energy dispersive X-ray fluorescence, and zeta potential. The anti-
activity of AgNps was assessed by broth microdilution and spot assays. Effects of AgNps on cellular and molecular targets were assessed by monitoring the intracellular reactive oxygen species (ROS) production in the absence and presence of natural antioxidant, changes in surface morphology, cellular ultrastructure, membrane microenvironment, membrane fluidity, membrane ergosterol, and fatty acids.
Spherical AgNps (10-30 nm) showed minimum inhibitory concentration (minimum concentration required to inhibit the growth of 90% of organisms) at 40 μg/mL. Our results demonstrated that AgNps induced dose-dependent intracellular ROS which exerted antifungal effects; however, even scavenging ROS by antioxidant could not offer protection from AgNp mediated killing. Treatment with AgNps altered surface morphology, cellular ultrastructure, membrane microenvironment, membrane fluidity, ergosterol content, and fatty acid composition, especially oleic acid.
To summarize, AgNps affected multiple cellular targets crucial for drug resistance and pathogenicity in the fungal cells. The study revealed new cellular targets of AgNps which include fatty acids like oleic acid, vital for hyphal morphogenesis (a pathogenic trait of
). Yeast to hypha transition being pivotal for virulence and biofilm formation, targeting virulence might emerge as a new paradigm for developing nano silver-based therapy for clinical applications in fungal therapeutics.
Silver nanoparticles (AgNps) have attracted maximal attention among all metal nanoparticles, and the study of their biological properties has gained impetus for further medical adoption. This study ...evaluated the cellular and molecular mechanisms associated with the action of AgNps against an opportunistic pathogen,
. Spherical, stable AgNp (average size 21.6 nm) prepared by a chemical reduction method showed minimum inhibitory concentration (required to inhibit the growth of 90% of organisms) at 40 μg/mL. AgNps have been reported to induce oxidative stress-mediated programmed cell death through the accumulation of intracellular reactive oxygen species (ROS). However, this study demonstrated that intracellular levels of AgNp-induced ROS could be reversed by using antioxidant ascorbic acid, but the sensitivity of AgNp-treated
cells could not be completely reversed. Moreover, in addition to the generation of ROS, the AgNps were found to affect other cellular targets resulting in altered membrane fluidity, membrane microenvironment, ergosterol content, cellular morphology, and ultrastructure. Thus, the generation of ROS does not seem to be the sole major cause of AgNp-mediated cell toxicity in
. Rather, the multitargeted action of AgNps, generation of ROS, alterations in ergosterol content, and membrane fluidity together seem to have potentiated anti-
action. Thus, this "nano-based drug therapy" is likely to favor broad-spectrum activity, multiple cellular targets, and minimum host toxicity. AgNps, therefore, appear to have the potential to address the challenges in multidrug resistance and fungal therapeutics.
Rhizospheric and non rhizospheric soil samples were taken from the tailings of North Eastern coal mines of Assam state of India to isolate the arsenic resistant bacteria having the ability to ...transform different arsenic forms particularly the toxic As (III) and As (V) using arsenic bioremediation methods. We describe the isolation and characterization of four strains with high resistance towards arsenic. Isolates were processed and characterized using biochemical and molecular characterization techniques. The 16S ribosomal RNA gene isolated were sequenced and the sequencing data were aligned using the multiple sequencing alignment tool (MSA). The sequences of the strains formed a monophyletic clade with the members of the genus Bacillus and Rhodanobacter in the The results of phylogenetic tree. The sequence representing Bacillus subtilis showed maximum similarity with 99% 16S rRNA gene sequence similarity followed by Bacillus cereus (99%) and Rhodanobacter thiooxydans with 97% similarity. The phylogenetic tree data suggests that the isolated and identified strains is of the type strains of species belonging to the genus Bacillus and Rhodanobacter together with the 16S rRNA gene sequence in the Ribosomal Database, small subunit rRNA and large subunit rRNA databases. It is of interest to isolate, identify and characterize the highly resistant strains, which were highly resistant to arsenic from the rhizospheric and non-rhizospheric soils from the tailings of the coalmines of Assam using the 16S rRNA gene sequence phylogenetic analysis. Thus, the four arsenic resistant strains were reported using the 16S rRNA gene sequence phylogenetic analysis. The accession numbers for the sequences are MN706211, MN701565, MN706210 and MN706212. A monophyletic clade representing the members of the genus Bacillus (Bacillus subtilis and Bacillus cereus) and Rhodanobacter thiooxydans is thus documented.
Processed food products can be contaminated by a variety of pathogenic and spoilage microbes. Those are the causative agents of foodborne diseases and cause significant economic losses in the food ...industries and human health. This was the basis of the present study, carried out with an objective to investigate the antibacterial and antifungal potentials of Mangifera indica bark extract. The antimicrobial activity of plant extracts and phytochemicals was evaluated with antibiotic susceptible and resistant microorganisms. An attempt was made in this study to evaluate these properties to prolong the shelf life of fruits and vegetables at the time of transportation and storage. Parts of the Mango plants have multifarious uses as traditional and asthetic values as well as medicinal implications. Antioxidant property was also evaluated by DPPH method. Methanolic extract of mangifera bark was used to evaluate the properties of phytochemicals and their role as an effective antioxidant and antimicrobial agent. The antibacterial activity was tested against one Gram-positive-Staphylococcus aureus, and three Gram-negative-Escherichia coli, Pseudomonas aeruginosa, Salmonella human pathogenic bacteria; and two fungal strains― Candida albicans and Aspergilus niger. After evaluation, methanolic extract of Mangifera bark the presence of phytochemicals like, alkaloids, glycosides, phenolic compounds, tannins and steroids. Zone of inhibition of extracts were compared with that of different standards like oxytetracycline, levofloxacin for antibacterial activity and fluconazole for antifungal activity. The results showed that the remarkable zone of inhibition against the bacterial growth ranged from 14 to 15 mm for pathogenic strain of bacteria. The zone of inhibition was observed first time of around 13 mm against the fungal strain. After the preliminary screening for antimicrobial activity by agar well diffusion method, MIC & MBC were also determined against above mentioned bacteria ranged from 3.125 to 6.25 μg/mL. However MBC ranges from 1.562 to 3.125 μg/mL. The parameter IC50 (efficient concentration|| value) is used for the interpretation of the results from the DPPH method and the IC50 value for Mangifera indica is 94.04 μg/ml. Therefore, this plant can be used to isolate bioactive natural products and can be employed to control the food spoilage and to improve the quality of fruits and food products and may also play a vital role for the improvement of quality products for food industries. Keywords: Mango bark, Antioxidant, Antimicrobial, hytochemicals, IC50, MIC, MBC
This work investigates solar still of double slope type included with N similar CPC integrated ETC having a series connection for sensitivity analysis using one at a time method by incorporating ...yield and efficiency. The energy-based analysis has been done by feeding all expressions of different parameters to computer code written in MATLAB-2015a for May month of New Delhi climatic situation. Fresh water yielding and energy efficiency values have been estimated for dissimilarities of mass flow rate maintaining all other parameters at the input of system constant and this procedure has been repeated till estimation is completed for all parameters at the input of system. All results so obtained have been 2D plotted. The fresh water yielding and energy efficiency for the considered system has been estimated to be most sensitive with respect to concentration ratio having mean sensitivity figure values as 0.91 and 0.78 in that order. Sensible conclusions have been presented based on the current analysis.
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