Antimicrobial and antioxidant activities of salt stress callus of Brinjal (Solanum melongena L.)

Ethanolic and methanolic salt stress callus extracts of Solanum melongena L. were tested for in vitro antimicrobial and free radical scavenging assayssuch as DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS+(2,2\u27Azinobis (3-ethyl benzo-thizoline-6-sulfonic acid) . In both the extracts the zone of inhibition is higher in Escherichia coli, Klebsiella pneumonia, Staphylococcus aureusand Streptococcus pyogenesat 90 µl concentration against the control. The antifungal activity of these extracts also the zone of inhibition is higher at 90 µl concentration against the control. The DPPH activity of different concentration of solvent extracts (1 mg/ml to 5 mg/ml) along with standard ascorbic acid among the five different concentration (50 µg/ml to 250 µg/ml) of extracts tested, the higher percentage of inhibition was observed in 250 µg/ml of methanol extract followed by ethanolic extract against the standard ascorbic acid. In ABTS+ activity the absorbance was increased with the increasing concentrations of both methanolic and ethanolic callus extracts

Semicontinuous Bioreactor Production of Recombinant Butyrylcholinesterase in Transgenic Rice Cell Suspension Cultures.

An active and tetrameric form of recombinant butyrylcholinesterase (BChE), a large and complex human enzyme, was produced via semicontinuous operation in a transgenic rice cell suspension culture. After transformation of rice callus and screening of transformants, the cultures were scaled up from culture flask to a lab scale bioreactor. The bioreactor was operated through two phases each of growth and expression. The cells were able to produce BChE during both expression phases, with a maximum yield of 1.6 mg BChE/L of culture during the second expression phase. Cells successfully regrew during a 5-day growth phase. A combination of activity assays and Western blot analysis indicated production of an active and fully assembled tetramer of BChE

Expression, Purification, and Biophysical Characterization of a Secreted Anthrax Decoy Fusion Protein in Nicotiana benthamiana.

Anthrax toxin receptor-mediated drug development for blocking anthrax toxin action has received much attention in recent decades. In this study, we produced a secreted anthrax decoy fusion protein comprised of a portion of the human capillary morphogenesis gene-2 (CMG2) protein fused via a linker to the fragment crystallizable (Fc) domain of human immunoglobulin G1 in Nicotiana benthamiana plants using a transient expression system. Using the Cauliflower Mosaic Virus (CaMV) 35S promoter and co-expression with the p19 gene silencing suppressor, we were able to achieve a high level of recombinant CMG2-Fc-Apo (rCMG2-Fc-Apo) protein accumulation. Production kinetics were observed up to eight days post-infiltration, and maximum production of 826 mg/kg fresh leaf weight was observed on day six. Protein A affinity chromatography purification of the rCMG2-Fc-Apo protein from whole leaf extract and apoplast wash fluid showed the homodimeric form under non-reducing gel electrophoresis and mass spectrometry analysis confirmed the molecular integrity of the secreted protein. The N-glycosylation pattern of purified rCMG2-Fc-Apo protein was analysed; the major portion of N-glycans consists of complex type structures in both protein samples. The most abundant (>50%) N-glycan structure was GlcNAc₂(Xyl)Man₃(Fuc)GlcNAc₂ in rCMG2-Fc-Apo recovered from whole leaf extract and apoplast wash fluid. High mannose N-glycan structures were not detected in the apoplast wash fluid preparation, which confirmed the protein secretion. Altogether, these findings demonstrate that high-level production of rCMG2-Fc-Apo can be achieved by transient production in Nicotiana benthamiana plants with apoplast targeting

A Study on Lapsation of Insurance Polices of Select Insurance Companies

Purpose: Lapsation is the discontinuation of payment of premia for reasons other than the death of a policy holder.  A majority of insurance companies notice that half of their policyholders do not pay their insurance premia regularly leading to an increase in lapsation of policies. Increase in lapsation not only affects the policyholders’ welfare but also insurance companies’ business growth. So, the present study was carried out to determine where lapsation rate is high and to offer solutions to reduce the rate of lapsation.   Design/Methodology/Approach: The present study is descriptive in nature. A carefully constructed questionnaire was used to gather the study's pertinent data. Data were collected from life insurance company employees in Tamilnadu using the convenience sampling method. The questionnaire includes inquiries about different policy kinds, distribution methods, agent information, and different commission structures.   Findings: Result of cross tabulation discloses that high rate of lapsation rate is noticed with term and endowment insurance policies both at LIC and private insurance companies.  High rate of lapsation is noticed under insurance policies distributed by direct sale teams of insurance companies. Further, it is noticed that lapsation rate is high in Private insurance companies, where insurance agents receive low commission. Lapsation rate is low at Public and Private Insurance companies, where insurance agents receive high and moderate level of commission.    Practical Implications: The result of the study will be immensely useful to policyholders in particular and insurance companies in general. Reduction in policy lapsation rate will assist policyholder to reap the benefit of availing insurance policies and aids life insurance companies to expand their business volume to a greater extent.   Social Implications: Life insurance companies not only safeguard interest of policyholders but also invests their excess amount of premium in corporate securities, thereby assists not only growth of industries but also Indian economy as well.  Lapsation of insurance policies may affect transfer of excess premium towards industry investment.  Thus, the present study assist to understand the reason for lapsation of policies and to offer suitable suggestions to reduce rate of lapsation and assist not only growth of insurance companies but also economy too.   Originality/Value: Questionnaire is employed for collecting first hand data. By contacting the life insurance company employees in person data required for the study has been gathered

Ultrasonic technology applied against mosquito larvae

The effective management of mosquito vectors is a timely challenge for medical and veterinary entomology. In this study, we evaluated the acoustic Larvasonic device to control young instars of the mosquito Aedes aegypti in diverse freshwater environments. Under laboratory conditions, we investigated the effect of exposure time and distance from the transducer on the mortality of larvae and pupae of Ae. aegypti. Furthermore, we evaluated the effectiveness of the ultrasound window of the electromagnetic spectrum under different field conditions. Results showed that first and second instar larvae were more sensitive to the frequency range of 18-30 kHz of the Larvasonic device. Ultrasonic waves applied for 180 s at a frequency from 18 to 30 kHz caused 100% larval mortality at a distance of 60 cm from the transducer. No mortality was observed in the non-target copepod Megacyclops formosanus. The exposure to the soundwaves produced by the acoustic larvicidal device over different distances effectively damaged Ae. aegypti through destruction of the larval dorsal tracheal trunk, thorax and abdomen. Overall, results indicated that the Larvasonic device tested can provide an alternative tool to reduce young instar populations of Ae. aegypti, without any effects on non-target aquatic invertebrates like copepods. It turned out to be a useful device for mosquito biocontrol. This technology has a relevant potential to fight the spread of mosquito-borne diseases

Facile synthesis of silver nanoparticles, anti-inflammatory, antibacterial and photocatalytic activities using Pogostemon speciosus Benth. An endemic medicinal plant

The development of biologically enthused green synthesis of silver nanoparticles (SNPs) has concerned significant global awareness about medical science and disease treatment. This paper discusses the green synthesis of SNPs using organic green sources; here we report a facile bottom-up ‘green’ route for the synthesis of SNPs using aqueous leaves extract of Pogostemon speciosus (Benth.) and evaluate its in-vitro anti-inflammatory, antibacterial and photocatalytic activities. The nanoparticles were investigated for the preparation of denaturation particles with PSLASNPs and the evaluation of anti-inflammatory activity with Protein denaturation and HRBC stabilization assays. Later, these PSLASNPs were studied for their potential role in antibacterial activity by well diffusion method, and Photocatalytic activity on degradation of dyes was demonstrated by using dyes Crystal violet, Coomassie blue, and Congo red. At 1000 µg/ml, the PSLASNPs have the greatest prevention of protein denaturation (71.92±1.37%), whilst the stabilization of the HRBC membrane exhibited significant anti-inflammatory action (64.39±1.61 %). The PSLASNPs showed the best antibacterial activity at the concentration of 10 µg/ml against Bacillus subtilis (8.2 mm), followed by Pseudomonas stuberia (6.2 mm) and Escherichia coli (6.4 mm), Staphylococcus aureus (5.3 mm), Staphylococcus gallinarium (4.5 mm) respectively at the same concentrations. Crystal violet, Coomassie blue, and Congo red were used for Photocatalytic activity on the breakdown of dyes. After 35 minutes, the degradation process was determined to be complete by the transformation of the reaction mixture's color to colorless. As a result, the PSLASNPs have anti-inflammatory, antibacterial, and photocatalytic activities

Transient Expression of Tetrameric Recombinant Human Butyrylcholinesterase in Nicotiana benthamiana.

To optimize the expression, extraction and purification of plant-derived tetrameric recombinant human butyrylcholinesterase (prBChE), we describe the development and use of plant viral amplicon-based gene expression system; Tobacco Mosaic Virus (TMV) RNA-based overexpression vector (TRBO) to express enzymatically active FLAG-tagged plant made recombinant butyrylcholinesterase (rBChE) in Nicotiana benthamiana leaves using transient agroinfiltration. Two gene expression cassettes were designed to express the recombinant protein in either the ER or to the apoplastic compartment. Leaf homogenization was used to isolate ER-retained recombinant butyrylcholinesterase (prBChE-ER) while apoplast-targeted rBChE was isolated by either leaf homogenization (prBChE) or vacuum-extraction of apoplastic wash fluid (prBChE-AWF). rBChE from apoplast wash fluid had a higher specific activity but lower enzyme yield than leaf homogenate. To optimize the isolation and purification of total recombinant protein from leaf homogenates, an acidic extraction buffer was used. The acidic extraction buffer yielded >95% enzymatically active tetrameric rBChE as verified by Coomassie stained and native gel electrophoresis. Furthermore, when compared to human butyrylcholinesterase, the prBChE was found to be similar in terms of tetramerization and enzyme kinetics. The N-linked glycan profile of purified prBChE-ER was found to be mostly high mannose structures while the N-linked glycans on prBChE-AWF were primarily complex. The glycan profile of the prBChE leaf homogenates showed a mixture of high mannose, complex and paucimannose type N-glycans. These findings demonstrate the ability of plants to produce rBChE that is enzymatically active and whose oligomeric state is comparable to mammalian butyrylcholinesterase. The process of plant made rBChE tetramerization and strategies for improving its pharmacokinetics properties are also discussed