Synthesis of Fluorophore Decorated Single-Walled Carbon Nanotubes for In-Vitro Cytocompatibility.

We report rhodamine based fluorophore derivative for the stable dispersion of single-walled carbon nanotubes (SWCNTs), which can afford better fluorescent label to carbon nanotubes (CNTs). The nanotubes-fluorophore conjugates are helpful in achieving stable dispersion in polar and non-polar solvents with intense fluorescence. The product was characterized through NMR, Mass spectrometry, Raman, XPS, SEM, AFM and Fluorescence measurements. The formation of SWCNT-g- Rhodamine was confirmed by the presence of D and G bands in Raman spectrum. The alkyl and aryl groups in the range of 14.8, 17.6, 38.1 and 96.3 ppm confirms the grafting of the nanocomposite through NMR. The morphological studies were carried out intensively for analyzing SWCNTs stable dispersion and the results from EDAX measurements shows the elements weight% of C: 35.09 and N: 30.1 concludes that SWCNTs are completely grafted onto rhodamine derivatives. The application of SWCNTs fluorophore conjugates were analyzed by cell viability studies using MTT assay and exhibits less toxic compare to other functionalized CNTs. The viability of percentage increases with decrease in the concentration of SWCNT-COCl with 91.7% of live cells even after 24 h at a concentration of 250 µg for SWCNT-g-Rhodamine. The fluorescent images obtained during viability analysis shows enhanced fluorescence for living cells in case of SWCNT-g-Rhodamine compared to SWCNT-COCl, which clearly shows the utility of decorating nanotubes with fluorophore. This research work further extends its application for molecular sensing and other biological process.

Cr-induced cellular injury and necrosis in Glycine max L.: Biochemical mechanism of oxidative damage in chloroplast.

Chromium-induced toxicity and mechanisms of cell death involved in plants are yet to be fully elucidated. To understand the events of these processes, the stress response of the soybean plant using trivalent and hexavalent chromium compounds, namely, basic chromium sulphate (BCS) and potassium dichromate (K2Cr2O7) was investigated. The leaf surface morphology for stomatal aperture, wax deposition and presence of trichomes for chromium accumulation was examined by SEM-EDAX and light microscopy. The leaf mesophyll cell integrity was identified by trypan blue staining; chlorophyll autofluorescence, ROS generation and mitochondrial function were studied by fluorescence microscopy using different dyes. Isolated chloroplasts were analysed for micronutrients and total chromium content by AAS. Elevated Cr level and decreased Fe, Cu and Zn content in chloroplast revealed the active transportation of highly soluble Cr6+ species resulting in poor absorption of micronutrients. Cr accumulation as Cr(V) in chloroplast was noticed at g = 1.98 of electron paramagnetic resonance signal. Plants grown in Cr(VI) amended soil showed chemical modification of biological macromolecules in the chloroplast as observed from fourier transform infra-red (FTIR) spectra; the chloroplast DNA damage was confirmed by DAPI staining. Cr(VI)-treated plants showed significant reduction in the levels of various biochemical parameters. The results altogether clearly indicate that Cr(VI)-induced reactive oxygen species (ROS) production leads to oxidative stress-associated changes in the organelles, particularly in chloroplast, resulting in cell death.

In vitro secretion of zymogens by bovine pancreatic acini and ultra-structural analysis of exocytosis.

The aim of this study is to establish a bovine pancreatic acinar cell culture model with longer viability and functionality. The cells could be maintained in a functional state for upto 20 days with normal morphology. Cells were positive for amylase as observed by immunofluorescence staining. Acinar cells are spherical and range about 2-3 µm in diameter. The porosome formed by exocytosis and heterogenous enzyme granules of size ranging 100-300 nm were seen on the surface of cells by electron microscopy. The activity of the enzymes was high on day 15 and the activity profile of the enzymes is in the order: protease>lipase>amylase and the enzymes were identified by SDS-PAGE. Long-term culture of bovine pancreatic acini could be useful in studying the pathogenesis of pancreatitis. Since the bovine genome shares about 80% identity with the human genome, the cells derived from bovine pancreas can be engineered and used as a potential xenotransplant to treat conditions like pancreatitis as the tissue source is abundantly available.

In vitro anti inflammatory activity of Aloe vera by down regulation of MMP-9 in peripheral blood mononuclear cells.

AIM OF THE STUDY: The anti-inflammatory activity of Aloe vera was investigated through MMP inhibition studies. The effect of Aloe vera on MMP-9 inhibition was tested on peripheral blood mononuclear cells (PBMC). MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from the heparinised venous blood by Ficoll Diatrizoate gradient centrifugation. The cell count and viability was determined using dye exclusion technique. Cytotoxicity was evaluated by MTT assay. Activation of MMP-9 was visualized by gelatin zymography. Inhibition of MMP-9 in the presence of aqueous extract of Aloe vera was detected by gelatin zymography and confirmed by RT-PCR. RESULTS: Peripheral blood mononuclear cells (PBMC) showed significant inhibition in the activity of MMP-9, indicating the in vitro inhibitory effect of Aloe vera on MMP-9. Zymographic analysis and RT-PCR showed that it caused a significant reduction in the production of MMP-9 in a concentration dependent manner. CONCLUSION: The inhibition of MMP-9 production in the cells was detected by gelatin zymography and was confirmed by RT-PCR.