Omega-3 fatty acids concentrate production by enzyme-catalyzed ethanolysis of supercritical CO<Subscript>2</Subscript> extracted oyster oil

The separation of oil by a suitable technique from the Pacific oyster muscle is important for the utilization of the oil as a ω-3 polyunsaturated fatty acids (ω-3 PUFAs) source and production of bio-functional peptides/ oligosaccharides from oil-free residue. This study was conducted to prepare ω-3 PUFAs concentrate from supercritical carbon dioxide (SC-CO₂) extracted Pacific oyster oil by enzyme-catalyzed ethanolysis reactions. SC-CO₂ extractions were done at different temperatures and pressures to optimize suitable extraction conditions and extracted oils were compared with Soxhlet (n-hexane) extracted oil to evaluate the yield and quality. Oil extracted by SC-CO₂ at optimized conditions was used for ethanolysis reaction catalyzed by immobilized sn-1,3 specific lipases, namely Novozymes-435, Lipozyme TLIM, and Lipozyme RMIM to produce 2-monoacylglycerols (2-MAG) rich in ω-3 PUFAs. The optimum temperature and pressure for SC-CO₂ extractions of oyster oil was 50°C and 30 MPa. In this condition, the yield of oil was 5.96% and the acid, peroxide, free fatty acid, and p-anisidine values were 4.49 mg KOH/g, 4.72 meq/kg, 3.42%, and 10.03, respectively. The ω-3 PUFAs content significantly increased in 2-MAG obtained from Novozymes 435, Lipozyme TLIM, and Lipozyme RMIM to 43.03 ± 0.36, 45.95 ± 0.29, and 40.50 ± 0.77%, respectively (p < 0.05). A thin layer chromatography (TLC) analysis confirmed the production and separation of 2-MAG in the ethanolysis process. The ratio of total ω-3 to ω-6 fatty acids was almost twice in 2-MAG of SC-CO₂ extracted oyster oil. SC-CO₂ extracted Pacific oyster oil can be used for sn-1,3 specific lipases catalyzed ethanolysis to produce ω-3 PUFAs rich in 2-MAG.     

Characterization of laser produced plasma using laser induced breakdown spectroscopy

The plasma parameter studies of the Nd:YAG (neodymium-doped yttrium aluminum garnet, Nd:Y₃Al₁₅O₁₂) crystal by using the fundamental (1064 nm) and second (532 nm) harmonics of Nd:YAG laser are reported. The electron temperature (T _e ) and electron number density (N _e) were determined using the Boltzmann plot method and the Stark-broadened line profile, respectively. An increase in the plasma parameters have been observed with an increase in the laser irradiance for both laser modes. The electron temperatures were calculated in the range of 0.53–0.66 eV for 1064 nm and 0.47–0.60 eV for 532 nm, and the electron number densities were determined in the range of 7.43 × 10¹⁵–3.27 × 10¹⁶ cm^?3 for 1064 nm and 1.35 × 10¹⁶–3.97 × 10¹⁶ cm^?3 for 532 nm in the studied irradiance range of 1.19–12.5 GW/cm². However, the spatial evolution of the plasma parameters investigated up to 2.75 mm away from the target surface at a fixed laser irradiance of 6.51 GW/cm2 showed a decreasing trend. In addition, the estimated values of the inverse bremsstrahlung (IB) absorption coefficients at both laser wavelengths showed that the IB process is dominant for the 1064-nm laser.     

Effect of reverse photoperiod on in vitro regeneration and piperine production in Piper nigrum L.

In this study, a novel approach for in vitro regeneration of Piper nigrum L. has been applied in order to increase healthy biomass, phytochemicals and piperine production via reverse photoperiod (16hD/8hL). Leaf portions of the seed-derived plants were placed on an MS-medium fortified with different PGRs. Under 16hD/8hL, thidiazuron (TDZ; 4.0 mg L⁻¹) and BA (1.5 mg L⁻¹) was found to be the most effective (< 90%) in callus induction. Two concentrations (1.5, 2.0 mg L⁻¹) of the IBA produced > 80% shoots from callus cultures. Healthy shoots were transferred to rooting medium and higher percentage of rooting (< 90%) was observed on IBA (1.5 mg L⁻¹). These in vitro tissues were subjected to amino acid analysis, spectrophotometry, and HPLC. ARG, SER, THR, and TYR were the most abundant components out of 17 amino acids. Higher amino acid production was observed under normal photoperiod (16hL/8hD) than under reverse photoperiod (16hD/8hL). The highest total phenolic content (TPC; 9.91 mg/g-DW) and flavonoid content (7.38 mg/g-DW) were observed in callus cultures incubated under 16hL/8hD than other tissues incubated under 16hD/8hL photoperiod. Higher DPPH and PoMo activities were observed in tissues incubated under 16hL/8hD photoperiod, while ABTS and Fe²⁺ chelating activities were found higher in tissues incubated under reverse photoperiod. Significant quantities of piperine content were observed in all tissues except callus cultures. These results suggest that reverse photoperiod is a promising approach for callus induction, phytochemicals and piperine production for commercial applications.     

Agrobacterium-Mediated Transformation of Tomato with rolB Gene Results in Enhancement of Fruit Quality and Foliar Resistance against Fungal Pathogens

Tomato (Solanum lycopersicum L.) is the second most important cultivated crop next to potato, worldwide. Tomato serves as an important source of antioxidants in human diet. Alternaria solani and Fusarium oxysporum cause early blight and vascular wilt of tomato, respectively, resulting in severe crop losses. The foremost objective of the present study was to generate transgenic tomato plants with rolB gene and evaluate its effect on plant morphology, nutritional contents, yield and resistance against fungal infection. Tomato cv. Rio Grande was transformed via Agrobacterium tumefaciens harbouring rolB gene of Agrobacterium rhizogenes. rolB. Biochemical analyses showed considerable improvement in nutritional quality of transgenic tomato fruits as indicated by 62% increase in lycopene content, 225% in ascorbic acid content, 58% in total phenolics and 26% in free radical scavenging activity. Furthermore, rolB gene significantly improved the defence response of leaves of transgenic plants against two pathogenic fungal strains A. solani and F. oxysporum. Contrarily, transformed plants exhibited altered morphology and reduced fruit yield. In conclusion, rolB gene from A. rhizogenes can be used to generate transgenic tomato with increased nutritional contents of fruits as well as improved foliar tolerance against fungal pathogens.     

Enhanced Phytoremediation of Cadmium-Contaminated Soil by Parthenium hysterophorus Plant: Effect of Gibberellic Acid (GA3) and Synthetic Chelator,Alone and in Combinations

The roles of gibberellic acid (GA₃) and ethylenediaminetetraacetic acid (EDTA) in phytoremediation of cadmium (Cd)-contaminated soil by Parthenium hysterophorus plant was investigated. GA₃ (10^?9, 10^?7, and 10^?5M) was applied as a foliar spray. EDTA was added to soil in a single dose (160 mg/kg soil) and split doses (40 mg/kg soil, four split doses). GA₃ and EDTA were used separately and in various combinations. P. hysterophorus was selected due to its fast growth and unpalatable nature to herbivores to reduce the entrance of metal into the food chain. The Cd phytoextraction potential of the P. hysterophorus plant was evaluated for the first time. Cd significantly reduced plant growth and dry biomass (DBM). GA₃ alone increased the plant growth and biomass in Cd-contaminated soil, whereas EDTA reduced it. GA₃ in combination with EDTA significantly increased the growth and biomass. The highest significant DBM was found in treatment T3 (10^?5M GA₃). All treatments of GA₃ or EDTA significantly enhanced the plant Cd uptake and accumulation compared with control (C1). The highest significant root and stem Cd concentrations were found in the combination treatment T11 (GA₃ 10^?5M + EDTA split doses), whereas in leaves it was found in the EDTA treatments. Cd concentration in plant parts increased in the order of stem < leaves < roots. The combination treatment T9 (GA₃ 10^?7M + EDTA split doses) showed the significantly highest total Cd accumulation (8 times greater than control C1, i.e., only Cd used). The GA₃ treatments accumulated more than 50% of the total Cd in the roots, whereas the EDTA treatments showed more than 50% in the leaves. Root dry biomass showed a positive and significant correlation with Cd accumulation. GA₃ is environment friendly as compared with EDTA. Therefore, further investigation of GA₃ is recommended for phytoremediation research for the remediation of metal-contaminated soil.     

Microbiological treatment of industrial wastes containing toxic chromium involving successive use of bacteria,yeast and algae

Bacteria, yeasts, protozoa and algae, observed in industrial effluents from tanneries, were checked for their possible use in the detoxification of polluted water. Two bacterial strains were found to be highly resistant to Cr^VI. Several bacteria, yeasts and algae were observed to be capable of reducing Cr^VI to Cr^III.     

Experimental infection of rhesus monkeys with Entamoeba histolytica mimics human infection

Experimental infection of Entamoeba histolytica was successfully established in rhesus monkeys (Macaca mulatta). Parasites of proven pathogenicity maintained through liver passage in hamsters were used in this study. Laparotomized monkeys were inoculated intracecally/intrahepatically with trophozoites, or by oral administration of cysts (2000 cysts/ml). The cysts were isolated from an infected human stool sample. Formation of typical amebic lesions in the inoculated tissues along with alterations in hematologic indices were studied in the infected monkeys. All the experimentally inoculated monkeys showed leukocytosis and mild neutrophilia, while the hemoglobin content and levels of blood sugar and blood urea remained unaltered during the post-infection period. Specific antiamebic antibodies were readily detectable in post-infected sera.     

Comparative evaluation of extraction methods for total proteins from Crocus sativus L. (Saffron)

Broadly speaking proteomic studies are one of the various techniques of utmost importance for understanding complex biological processes that occur under inductive conditions and revealing the multidimensional aspects of Crocus sativus in biological systems. In order to get an insight into the molecular changes and to characterize the variations in protein expression of C. sativus, a detailed proteomic analysis on one-dimensional gel electrophoresis is one of the basic steps to accomplish. We have compared total protein profiles of C. sativus extracted by three different recipes and analyzed on 10% sodium dodecyl sulfate polyacrylamide gels. Gels were subjected to densitometric analysis for further characterization. Among three different protocols NP-40 extraction buffer recipe resulted in the extraction of proteins most efficiently with minimum background and streaking. There was maximum solubilization of proteins with high efficiency. Such a profile can be used for high precision analysis of differential protein expression. This work is an attempt to assist researchers in effective extraction of proteins from C. sativus. As a researcher faces a perplexing array of choices as where to start we describe a method based on our collective analysis of the different protein protocols. This paper presents a method that could be applied at the outset of any proteomic study.     

In vitro label‐free screening of chemotherapeutic drugs using Raman microspectroscopy: Towards a new paradigm of spectralomics

This overview groups some of the recent studies highlighting the potential application of Raman microspectroscopy as an analytical technique in preclinical development to predict drug mechanism of action and in clinical application as a companion diagnostic and in personalised therapy due to its capacity to predict cellular resistance and therefore to optimise chemotherapeutic treatment efficacy. Notably, the anthracyclines, doxorubicin and actinomycin D, elicit similar spectroscopic signatures of subcellular interaction characteristic of the mode of action of intercalation. Although cisplatin and vincristine show markedly different signatures, at low exposure doses, their signatures at higher doses show marked similarities to those elicited by the intercalating anthracyclines, confirming that anticancer agents can have different modes of action with different spectroscopic signatures, depending on the dose. The study demonstrates that Raman microspectroscopy can elucidate subcellular transport and accumulation pathways of chemotherapeutic agents, characterise and fingerprint their mode of action, and potentially identify cell‐resistant strains. The consistency of the spectroscopic signatures for drugs of similar modes of action, in different cell lines, suggests that this fingerprint can be considered a “spectralome” of the drug‐cell interaction suggesting a new paradigm of representing spectroscopic responses.