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V Sahai

Publications and source records attributed to V Sahai.

13 recordsLinked to original sources

Enhanced expression of the recombinant lethal factor of Bacillus anthracis by Fed-Batch culture.

High cell density cultivation has been one of the most effective ways to increase cell as well as the product yields. The structural gene for the 90-kDa lethal factor (LF) isolated from Bacillus anthracis was expressed as fusion protein with 6x histidine residues under the transcriptional regulation of the T5 promoter in Escherichia coli. Various strategies were tried to scale up the expression of the recombinant lethal factor by bioprocess optimization using fed batch culture technique in a 14 litre fermentor. The media, a defined mixture of salts, trace elements, vitamins, etc. along with a specified carbon source was used for the growth. The pH of the media was maintained at 6.8 while the temperature was changed from 37 to 28 degrees C during the cultivation. During the growth and induction phases, the DO was maintained above 20% by automatic control of agitation. The specific growth rate was controlled by utilizing an exponential feeding profile determined from mass balance equations. As a result of control of specific growth rate at two different levels, there was about twenty five fold increase in biomass compared to the biomass in the shake flask. E. coli cells yielded a soluble cytosolic protein with an apparent molecular mass of 90 kDa. The protein was purified to homogeneity using metal chelate affinity chromatography, followed by anion exchange on FPLC using Mono-Q column. In solution, trypsin cleaved protective antigen bound to native and recombinant LF with comparable affinity. The recombinant LF resembled the LF purified from B. anthracis in the macrophage lysis assay, using a murine macrophage cell line J774A.1 sensitive to anthrax toxin. It was possible to achieve a yield of 50 mg of the purified protein from 1 litre culture broth.

Antigens, Bacterial↗

Task related changes in ERP auditory vs visual tasks.

P3 component of event related potentials have been studied in relaxed state of mind. Changes in P3 latency and amplitude are expected if measured after a rigorous mental exercise. The event related potentials (ERP) were measured in 20 normal male subjects aged between 18 and 22 years. ERP was recorded twice in each subject before and after the administration of a mental task. The auditory mental task comprised of repeating in reverse order string of random digits read out to the subject at a uniform speed of 1 per second. The visual task comprised reading laterally inverted type written text. Each subject had to undergo 2 sessions in separate sittings. The latency and amplitude of P300 recorded before and after the task were compared statistically using Students' unpaired t-test. There was significant increase in P3 latency following both visual and auditory tasks (auditory task: before 330 +/- 24.43 msec and after 342.9 +/- 27.66 msec, similarly visual task: before 333.2 +/- 26.66 msec and after 345.65 +/- 26.56 msec). All other ERP waves latency and amplitude changes were insignificant.

Acoustic Stimulation↗

Task related changes in contingent negative variation (CNV) response of endogenous evoked potentials.

Contingent negative variation (CNV) is a slow negative cortical potential shift, which occurs during a warned foreperiod reaction time paradigm. Most studies of evoked potentials have concentrated on components occurring during the first 300 msec, although there are important and recordable aspects of signal processing occurring well beyond 300 msec e.g. late negative slow wave. CNV has proven sensitive to a number of psychological variables, none of which can yet be singled out as a definitive or exclusive correlate. Changes are expected if measured after a rigorous mental exercise. CNV was measured in 20 normal male subjects aged between 18 and 20 years. CNV was recorded twice in each subject before and after the administration of a mental task. The auditory mental task comprised repeating in reverse order string of random digits read out to the subject at a uniform speed of 1 per second. The visual mental task comprised reading laterally inverted type written text. Each subject had to undergo 2 sessions in separate sittings. The latency and amplitude of waves N1, P3, orientation (O), expectancy (E), CNV, reaction time (RT) were recorded. These values recorded before and after the task were compared statistically using student's unpaired t-test. The significant latencies recorded before and after the auditory task were, N1: 88.00 +/- 11.96 and 100.00 +/- 21.52 msec, P3: 289.00 +/- 54.85 and 299.00 +/- 52.91 msec, reaction time (RT): 102 +/- 17.05 msec and 123 +/- 17.5 msec, and in case of visual task, N1: 88 +/- 13.16 msec and 99.00 +/- 16.51 msec, reaction time (RT): 107 +/- 11.74 msec and 127 +/- 13.42 msec respectively. All other CNV wave latencies and amplitude changes were insignificant. Hence task effects sensory perception as reflected by increased latency of the long latency response N1 and the cortical integrative processes resulting in increased reaction time.

Adolescent↗

Evaluation of biomass.

Evaluation of biomass concentration is an important problem encountered in many microbial and other bioprocesses. It determines the catalytic activity of the microbial cell in a given time. Various direct and indirect methods for the estimation of biomass have been developed using physical and biochemical techniques. Despite many promising classical methods available, the evaluation of microbial growth in bioprocesses may sometimes become laborious, impracticable and give erroneous values. Various methods for enumeration of organisms and determination of biomass, including recent developments in monitoring biomass concentration for the control of biotechnological processes, are discussed taking into the consideration their practical importance, usefulness and constraints in application.

Biotechnology↗

Production of cellulases by Trichoderma reesei QM 9414 in fed-batch and continuous-flow culture with cell recycle.

The scope in improving enzyme productivities from the cellulose fermentation process is examined in laboratory-scale fermentors. The maximum productivity (30 IU/liter hr) is attained in a continuous-culture process with cell recycle using modified medium containing 0.5% cellulose. Optimum dilution rate and recycle ratio are determined as 0.025 hr-1 and 1.2, respectively, for the process. The system is analyzed and steady-state equations for predicting enzyme protein concentrations in the fermentor are developed. In fed-batch cultures, slow addition of cellulose at high concentrations can improve enzyme productivity by as much as 33% over a batch process. The scope and results of using modified medium for cellulase production are also presented.

Cellulase↗

Predicting future long-term-care needs in a community.

Under current financial restraints, planning for the future long-term-care needs of older Canadians is coming under increasing scrutiny. Even though health care is moving to a community-based system (i.e., deinstitutionalization), this is not necessarily the most cost-effective strategy for severely disabled older adults. Three models are used in the present paper to estimate the number of severely disabled (two or more ADL disabilities abilities) older adults expected in the next 5 to 10 years in a small community in Northern Ontario. While mortality and disability rates are the main predictors, it was also necessary to consider additional factors particular to the community in question. It is suggested that for long-term-care planning to be effective and economically sound, similar strategies will be needed throughout Canada.

Activities of Daily Living↗