A novel study for producing complexed and encapsulated nutrients at nanometric scale to enhance plant growth.

Complexation of micronutrients with complexing agents reduce undesirable reactions of fertilizers in soil water system. In the form of complex structure nutrients remain available to plants in the useable form. Nanoform fertilizer enhances the surface area of particles and less amount of fertilizer contact with large area of plant roots which reduce fertilizer cost. Controlling release of fertilizer using polymeric material like sodium alginate makes agriculture practices more efficient and cost effective. Several fertilizers and nutrients are used at a large scale to improve crop yields globally and almost more than half goes to waste. Therefore, there is a dire need to improve plant-available nutrients in soil, using feasible, environmentally friendly technologies. In the present research, complexed micronutrients were successfully encapsulated using a novel technique at nanometric scale. The nutrients were complexed with proline and encapsulated using sodium alginate (polymer). Sweet basil was subjected to seven treatments over three months in a moderately controlled environment (25 °C of temperature and 57% of humidity) to study the effects of synthesized complexed micronutrient nano fertilizers. The structural modifications of the complexed micronutrient nanoforms of fertilizers were examined, through X-ray powder diffraction (XRD) and scanning electron microscopy (SEM). The size of manufactured fertilizers was between 1 and 200 nm. Fourier transform infrared (FTIR) spectroscopy stretching vibration peaks at 1600.9 cm-1 (C=O), 3336 cm-1 (N-H) and at 1090.2 cm-1 (N-H in a twisting and rocking) corresponds to the pyrrolidine ring. Gas chromatography-mass spectrometry was used to analyze the chemical makeup of the essential oil of the basil plants. Essential oil yield of basil plants increased from 0.0035 to 0.1226% after treatments. The findings of the present research show that complexation and encapsulation improve crop quality, essential oil yield, and antioxidant potential of basil.

Surface-enhanced Raman spectroscopy for the characterization of different anatomical subtypes of oral cavity cancer.

Background The prognosis for oral cancer patients is still very poor worldwide. Early detection and treatment therapy remain the key issue to be addressed for improved patient survival. The characteristic Raman spectral features associated with the biochemical changes in the blood serum samples can be used for the diagnosis of diseases, particularly for oral cancer. Surface-enhanced Raman spectroscopy (SERS) is a promising technique for non-invasive and early detection of oral cancer by analyzing molecular changes in body fluids. Objectives To detect oral cavity anatomical subsites (buccal mucosa, cheek, hard palate, lips, mandible, maxilla, tongue and tonsillar region) cancers by using blood serum samples, SERS with principal component analysis is used. Material and Method SERS is employed with silver nanoparticles for the analysis and detection of oral cancer serum samples by comparing with healthy serum samples. SERS spectra are recorded by Raman instrument and preprocessed using the statistical tool. Principal component analysis (PCA) and Partial least square discriminant analysis (PLS-DA) are used to discriminate between oral cancer serum samples and control serum samples. Results Some major SERS peaks are observed at 1136 cm-1 (Phospholipids) and 1006 cm-1 (Phenylalanine) remain higher in intensities for oral cancer spectra as compared to healthy spectra. The peak at 1241 cm-1 (amide III) is observed only in oral cancer serum samples while absent in healthy serum samples. Higher protein and DNA contents were detected in SERS mean spectra of oral cancer. Moreover, PCA is used to identify the biochemical differences in the form of SERS features which is used to differentiate between oral cancer and healthy blood serum samples, while PLS-DA is used to build differentiation model of oral cancer serum samples and healthy control serum samples. PLS-DA provides successful differentiation with 94% specificity and 95.5% sensitivity. Conclusions SERS can be used for the diagnosis of oral cancer and to identify metabolic changes that occur during disease development.

Antibiofilm and Quorum Sensing Inhibition (QSI) Potential of Lagerstroemia speciosa Leaves Extract.

Disruption of quorum sensing pathway of pathogenic microbes is considered as novel approach to fight against infectious diseases. The current study was planned to evaluate the antibiofilm and quorum sensing inhibitory potential of Lagerstroemia speciosa. Antibacterial and antibiofilm potential of L. speciosa extracts was determined through agar well diffusion and crystal violet assay against sinusitis isolates, that is, Staphylococcus aureus, Enterococcus faecalis, Proteus mirabilis, and Klebsiella pneumoniae, while quorum sensing inhibition efficacy of L. speciosa extracts was determined through violacein inhibition assay using Chromobacterium pseudoviolaceum as bacterial model. The methanolic extract of L. speciosa presented the highest antimicrobial activity against E. faecalis and antibiofilm activity against K. pneumoniae (77.42 ± 1.51%), while n-hexane extract was found to be least active against all tested bacterial strains. Quorum sensing inhibition activity of L. speciosa extracts against C. pseudoviolaceum showed significant dose-dependent inhibition in violacein production by different concentrations of methanolic extract. Furthermore, none of the extracts of L. speciosa showed any hemolytic activity against human RBCs and hold considerable thrombolytic potential in comparison to streptokinase (75.9 ± .46%). In conclusion, findings suggest that L. speciosa leaves are excellent source of phytochemicals with potent antibiofilm and quorum sensing inhibition potential.

Induction of Erythrocyte Shrinkage by Omeprazole.

Omeprazole, a proton pump inhibitor blocks the H+/K+-ATPase channels of gastric parietal cells. It is used for the treatment of peptic ulcer. Prolonged use of omeprazole may involve in inducing anemia. The key marker of eryptosis includes membrane blebbing, cell shrinkage and phosphatidylserine (PS) exposure at the cell surface. In current study, the eryptotic, oxidative as well as hemolytic effects of therapeutical doses (0.5, 1 and 1.5 µM) of omeprazole were investigated after exposing erythrocytes for 48 hours. Investigation of eryptosis was done by cell size measurement, PS exposure determination and calcium channel inhibition. As a possible mechanism of omeprazole induced eryptosis, oxidative stress was investigated by determining the catalase, glutathione peroxidase and superoxide dismutase activities. Similarly, necrotic effect of omeprazole on erythrocytes was also evaluated through hemolysis measurement. Results of our study illustrated that 1.5 µM of omeprazole may induce significant decrease in superoxide dismutase, glutathione peroxidase and catalase activities as well as triggered the erythrocytes shrinkage, PS exposure and hemolysis. Role of calcium was also confirmed in inducing erythrocyte shrinkage. It is concluded that the exposure of erythrocytes with 1.5 µM omeprazole may enhance the rate of eryptosis and hemolysis by inducing oxidative stress.

Cytotoxic, α-amylase inhibitory and thrombolytic activities of organic and aqueous extracts of Bacillus clausii KP10.

Humans are experiencing serious health issues like myocardial infarction and diabetes. Thrombosis is the reason of myocardial infarction that may cause death. Bioactive compounds or enzymes can be used to dissolve the clot. Whereas diabetes is a disorder of metabolism in which the level of glucose in blood becomes high. It can be controlled by inhibiting α-amylase enzyme. The current project was, therefore planned to investigate the thrombolytic, α-amylase inhibitory and cytotoxic (to access drug safety) potentials of the organic and aqueous bioactive fractions of Bacillus clausii KP10. The cytotoxicity was assessed with hemolytic assay, α-amylase inhibition assay was done by using DNS and in-vitro thrombolytic effect was checked with human blood. In our experiments, the maximum hemolytic activity was shown by ethyl acetate fraction (12.64%). Results were compared with standard Triton X-100 which showed 91.61% hemolytic activity whereas all other fractions showed least cytotoxic activity. The extracts were also evaluated as thrombolytic agents as correlated to streptokinase (73.83%). All the extracts showed clot lysis activity, among which water soluble fraction exhibited maximum (35.16%) clot lysis activity. In our experiment methanol soluble fraction of B. clausii KP10 showed maximum 26.49% α-amylase inhibitory activity. Results were analyzed statistically through analysis of variance (ANOVA).

Assessment of potential interaction between simvastatin and clarithromycin in healthy adult male subjects.

Cardiac patients with weak immune system are susceptible to bacterial infections. Their prescriptions frequently contain simvastatin and clarithromycin together. The objective of present project was to assess the potential interaction between simvastatin and clarithromycin by evaluating the clarithromycin effects on the pharmacokinetics of simvastatin in healthy adult male subjects. The study design comprised of two phases, used at interval of one week. In first phase simvastatin 20 mg alone was administered to each volunteer. In second phase, co-administration of simvastatin 20 mg with clarithromycin 250 mg was made under similar specified conditions. Blood samples were collected at specified time intervals. Simvastatin plasma concentrations were analyzed through High Performance Liquid Chromatography with UV detector at 238 nm wavelength. Using one compartment open model, MW/PHARM version 3.02 software program was used by F. Rombut for pharmacokinetic parameters calculation. Clarithromycin co-treatment resulted in 2.3 fold increase in maximum plasma concentration Cmax (from 2.47±0.34 ng.mL-1 to 5.66±1.18 ng.mL-1; p<0.05) and 3.9 fold increase in area under time versus concentration curve from 0 to 10 hours AUC0-10 (from 15.10±3.73 ng.hr.mL-1 to 58.49±15.73 ng.hr.mL-1; p<0.05) of simvastatin. These results suggest that co-prescription of simvastatin and clarithromycin should be avoided to minimize the adverse events resulting from high simvastatin concentration, without sacrificing therapeutic worth of simvastatin.

Sequence Analysis and Comparative Bioinformatics Study of Camelysin Gene (calY) Isolated from Bacillus thuringiensis.

Bacillus strains have been widely used for the production of fibrinolytic enzymes having role in the treatment of cardiovascular disorders. Purification and overproduction of such enzymes has increased their usage in medical fields including metalloproteinases with the ability to degrade extracellular matrix (ECM). Camelysin, a neutral metalloproteinase has been isolated from different species of bacteria like Bacillus cereus, Bacillus anthracis, and Bacillus thuringiensis with fibrinolytic, collagenolytic and actin degradation activity. This project successfully demonstrated the presence of 734-bp coding DNA sequence (CDS) encoding a 20.72331 kDa camelysin gene in local strain of Bacillus thuringiensis containing a signal peptide with cleavage site between residues 19 and 20. The sequence was submitted to GenBank (KT023597) and the sequence showed high homology with the camelysin protein of closely related Bacillus species. The alignment of related proteins through ClustalW displayed difference of four amino acids ("Q" replaced by "P" at position 169 and at position 182-184, "NQE" replaced by "HLK") in the isolated protein. Comparison including structural and functional analysis of camelysin sequences isolated from different Bacillus species was carried out using different bioinformatics tools and software. The information would help in better understanding the properties of camelysin protein and its role in pathogenicity and clinical treatments.

Synthesis of Some Unique Carbamate Derivatives bearing 2-Furoyl-1-piperazine as a Valuable Therapeutic Agents.

The aim of the research work was to synthesize different biologically active carbamate derivatives bearing 2-furoyl-1-piperazine and having modest toxicity. The synthesis was completed as a multiple sequence. The structural confirmation of all the synthesized compounds was obtained by EI-MS, IR and 1H-NMR spectral data. The enzyme inhibition and antibacterial potential of the synthesized compounds was evaluated. To find the utility of the prepared compounds as possible therapeutic agents their cytotoxicity was also checked. All the compounds were active against acetylcholinesterase enzyme, especially 12 and 14 showed very good inhibitory potential relative to Eserine, a reference standard. Almost all the compounds showed good activities against both Gram-positive and Gram-negative bacterial strains.

Recombinant Expression and Phenotypic Screening of a Bioactive Cyclotide Against α-Synuclein-Induced Cytotoxicity in Baker's Yeast.

We report for the first time the recombinant expression of fully folded bioactive cyclotides inside live yeast cells by using intracellular protein trans-splicing in combination with a highly efficient split-intein. This approach was successfully used to produce the naturally occurring cyclotide MCoTI-I and the engineered bioactive cyclotide MCoCP4. Cyclotide MCoCP4 was shown to reduce the toxicity of human α-synuclein in live yeast cells. Cyclotide MCoCP4 was selected by phenotypic screening from cells transformed with a mixture of plasmids encoding MCoCP4 and inactive cyclotide MCoTI-I in a ratio of 1:5×10(4). This demonstrates the potential for using yeast to perform phenotypic screening of genetically encoded cyclotide-based libraries in eukaryotic cells.