Squalene production in the cell suspension cultures of Indian sandalwood (<Emphasis Type="Italic">Santalum album</Emphasis> L.) in shake flasks and air lift bioreactor

Squalene has emerged as a specialty chemical being important in nutraceutical, pharmaceutical, vaccine and cosmetic industries due to the anticancer, antioxidant, skin hydrating, immune stimulating and emollient activities. The main source of squalene is shark liver oil, but alternate sources are being sought to secure sustainable supply of squalene and conserve the marine sources. The present work was carried out with the aim of generation of this compound in the cell suspension cultures of sandalwood, a medicinally important plant rich in sesquiterpenes. To achieve this, highly proliferating cell lines of sandalwood were generated by medium optimization. Accumulation of squalene in the cells was studied in a time dependent manner in shake flasks and bioreactor. In shake flask experiments, a significant amount of 3.2 mg/g dry weight was accumulated in 6 weeks of culture whereas the cells performed much better in bioreactor where squalene accumulation was found to be 5.5 mg/g dry weight in 4 weeks. Enzymes of lower terpenoid pathway namely hydroxy-3 methylglutaryl-coenzyme A reductase, farnesyl diphosphate synthase and squalene synthase showed a positive correlation with the product accumulation as evident from GC analysis of the reaction products and terpenoids accumulated. The findings of present work describe a high potential for commercial application since the marine sources have pollution concerns, vegetal sources are limited and the compound offers great hope in prevention of human chronic diseases like cancer. To our knowledge this is the first report about squalene generation in medicinally important sandalwood plant cell suspensions.     

Interpenetrating biopolymer network based hydrogels for an effective drug delivery system

Discovery of hydrogels has resulted in developing competent controlled-release drug delivery systems. Present study describes the synthesis and characterization of novel pH responsive hydrogels of chitosan, hydroxyl ethyl cellulose (HEC) and polyol prepared by physical blending of the three components in different ratios. Vegetable oil derived polyol seems to act as a filler and cross linking agent. The synthesized hydrogels were characterized using FT-IR spectroscopy, thermo gravimetric analysis (TGA), Optical microscopy and scanning electron microscopy (SEM). Equilibrium swelling behavior of hydrogels in water and different buffers with pH values (2, 4, 7.3, and 8) indicated the sustained expansion of the films in different pH solutions.     

Thermo-optic measurements and their inter-dependencies for delineating cancerous breast biopsy tissue from adjacent normal

The histopathological diagnosis of cancer is the current gold standard to differentiate normal from cancerous tissues. We propose a portable platform prototype to characterize the tissue's thermal and optical properties, and their inter-dependencies to potentially aid the pathologist in making an informed decision. The measurements were performed on 10 samples from five subjects, where the cancerous and adjacent normal were extracted from the same patient. It was observed that thermal conductivity (k) and reduced-scattering-coefficient (μ'_s) for both the cancerous and normal tissues reduced with the rise in tissue temperature. Comparing cancerous and adjacent normal tissue, the difference in k and μ'_s (at 940 nm) were statistically significant (p = 7.94e-3), while combining k and μ'_s achieved the highest statistical significance (6.74e-4). These preliminary results promise and support testing on a large number of samples for rapidly differentiating cancerous from adjacent normal tissues.     

Raman spectroscopy‐based detection of RNA viruses in saliva: A preliminary report

Several non‐invasive Raman spectroscopy‐based assays have been reported for rapid and sensitive detection of pathogens. We developed a novel statistical model for the detection of RNA viruses in saliva, based on an unbiased selection of a set of 65 Raman spectral features that mostly attribute to the RNA moieties, with a prediction accuracy of 91.6% (92.5% sensitivity and 88.8% specificity). Furthermore, to minimize variability and automate the downstream analysis of the Raman spectra, we developed a GUI‐based analytical tool “RNA Virus Detector (RVD).” This conceptual framework to detect RNA viruses in saliva could form the basis for field application of Raman Spectroscopy in managing viral outbreaks, such as the ongoing COVID‐19 pandemic. ( http://www.actrec.gov.in/pi-webpages/AmitDutt/RVD/RVD.html ).     

High quality protein microarray using in situ protein purification

Background  

In the postgenomic era, high throughput protein expression and protein microarray technologies have progressed markedly permitting screening of therapeutic reagents and discovery of novel protein functions. Hexa-histidine is one of the most commonly used fusion tags for protein expression due to its small size and convenient purification via immobilized metal ion affinity chromatography (IMAC). This purification process has been adapted to the protein microarray format, but the quality of in situ His-tagged protein purification on slides has not been systematically evaluated. We established methods to determine the level of purification of such proteins on metal chelate-modified slide surfaces. Optimized in situ purification of His-tagged recombinant proteins has the potential to become the new gold standard for cost-effective generation of high-quality and high-density protein microarrays.     

Effect of DDT on the free amino acid pool of brain and kidney in guinea-pigs.

Free amino acid composition of brain and kidney of guinea-pig showed significant decrease after repeated exposure of skin to P-P1 DDT. Gamma amino butyric acid of the brain and proline, tyrosine and valine of kidney disappeared completely. The remaining amino acids showed a marked depletion.