Severe Disease Burden and the Mitigation Strategy in the Arsenic-Exposed Population of Kaliprasad Village in Bhagalpur District of Bihar, India.

The present study was carried out in the village Kaliprasad of Bhagalpur district of Bihar to know the arsenic exposure effect in the exposed population. A total of n = 102 households were studied, and their water and biological samples such as urine and hair were collected and analyzed in a graphite furnace atomic absorption spectrophotometer (GF-AAS). The assessment of arsenic-exposed village population reveals that the villagers were suffering from serious health-related problems such as skin manifestations (hyperkeratosis and melanosis in their palm and soles), breathlessness, general body weakness, mental disorders, diabetes, hypertension (raised blood pressure), hormonal imbalance, neurological disorders, and few cancer cases. About 77% of household hand pump water had arsenic level more than the WHO recommended level of 10 µg/L, with highest level of 523 µg/L. Moreover, in 60% individual's urine samples, arsenic concentration was very high with maximum 374 µg/L while in hair 64% individuals had arsenic concentration above the permissible limit with maximum arsenic concentration of 11,398 µg/kg. The hazard quotient (HQ) was also calculated to know the arsenic risk percentage in children as 87.11%, in females as 83.15%, and in males as 82.27% by groundwater. This has surpassed the threshold value of 1 × 10 - 6 for carcinogenic risk (CR) in children, female, and male population group in the village. Hence, the exposed population of Kaliprasad village are at very high risk of the disease burden.

Comparison of Palonosetron Versus Palonosetron and Dexamethasone for Prevention of Postoperative Nausea and Vomiting After Middle Ear Surgeries: A Randomized Controlled Study.

Background: Postoperative nausea and vomiting (PONV) are one of the common distressing conditions after anesthesia. The PONV are related to several potential risk factors are patient related, anesthesia related, and surgery related. In surgery-related risk, middle ear surgery is associated with a high incidence of PONV. Aims: This study aimed to compare the efficiency of palonosetron versus palonosetron with dexamethasone in the prevention of PONV in middle ear surgeries. Settings and Design: This was a prospective, randomized, double-blind study. Statistical Analysis: The data were presented as descriptive statistics for continuous variables and percentages for categorical variables and were subjected to Z-test/Chi-square test/Fisher's exact test. The value of P < 0.05 was considered statistically significant. Results: Demographic parts in comparison to age, duration of surgery, and duration of anesthesia were similar in both the groups. Our study showed that the incidence of PONV during 0-6 h was 38% (n = 19) in Group A and 12% (n = 6) in Group B and the incidence during 6-12 h postoperatively was 14% (n = 7) in Group A and 8% (n = 4) in Group B. During 12-24 h, the incidence was 8% (n = 4) and 6% (n = 3) in Group A and B, respectively. Hence, the difference of total early PONV in Group A was 60% (n = 30) and in Group B, it was 26% (n = 13) which was statistically significant (P < 0.03). Conclusions: The above result proves that palonosetron and dexamethasone group is superior in the prevention of PONV in middle ear surgery.

Nutrient-depended metabolic switching during batch cultivation of Streptomyces coelicolor explored with absolute quantitative mass spectrometry-based metabolite profiling.

The well-known secondary metabolite-producing bacterium Streptomyces coelicolor is a natural choice for the development of super-hosts optimized for the heterologous expression of antibiotic biosynthetic gene clusters (BGCs). In this study, we used S. coelicolor M145 and its derivative strain M1146 where all active BGCs have been deleted and generated high-resolution quantitative time series metabolite profiles under two cultivation conditions (phosphate and nitrogen limitation to cease growth and trigger secondary metabolism). Five targeted LC-MS/MS-based methods were used to quantify intracellular primary metabolites covering phosphorylated metabolites, amino acids, organic acids, (deoxy) nucleoside/sugar phosphates, Nicotinamide adenine dinucleotide (NAD), and Coenzyme A (CoA). The nitrogen limitation resulted in a sharp decline in respiration and an immediate drop in the cell mass concentration. Intracellularly, a reduction in the level of the metabolites next to α-ketoglutarate in the tricarboxylic acid cycle and a decrease in the NADH pool were among the most prominent adaptation to this nutrient limitation. Phosphate limitation evoked a different adaptation of the metabolite pools as most of the phosphorylated metabolite pools except 6-phosphogluconic acid (6PG) pool were downregulated. 13C-isotope-labeling experiments revealed the simultaneous activity of both glycolysis and gluconeogenesis during the co-utilization of glucose and glutamate. The S. coelicolor M1146 strain had similar time-series metabolite profile dynamics as the parent M145 strain, except for a visibly increased 6PG pool in the stationary phase. In general, the nutrient limitation had a larger effect on the metabolite pool levels than the absence of secondary metabolite production in M1146. This study provides new insight into the primary carbon metabolism and its link to the secondary metabolism which is needed for further optimization of both super-host genotype and cultivation conditions. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-022-03146-x.

Monitoring lipids profile, CO2 fixation, and water recyclability for the economic viability of microalgae Chlorella vulgaris cultivation at different initial nitrogen.

The economic viability of microalgae as a bioenergy source depends on many factors. High CO2 fixing rate, improved lipids yield, and minimum water footprint are few key parameters. This study investigates the effect of four initial nitrogen concentrations (1-, 2-, 6- and 10-mM as nitrate) on lipids yield, their classification and composition, CO2 fixation rate, and water quality for further reuse after first cultivation. The initial 6 mM nitrate was found optimum for the growth and overall lipid productivity of Chlorella vulgaris. The maximum quantum efficiency (as Fv/Fm ratio) for algae decreases along with the cell growth profile and depletion of the initial nitrate concentration. CO2 fixation rate increased initially and peaked during exponential growth and then declined for the rest of the cultivation period. A higher CO2 fixation rate was recorded at 6 mM, and an overall fixation rate of CO2 was high at 6 mM. A higher total organic carbon (TOC) is produced in recycled water at a low nitrogen concentration of 1 and 2 mM. TOC changes during the cultivation period and with each reuse of water. Water was recycled twice successfully, while growth was inhibited during the 3rd cycle. Based on all these investigations, 6 mM of initial nitrogen was found optimal at given growth conditions.

A comparative study at bioprocess and metabolite levels of superhost strain Streptomyces coelicolor M1152 and its derivative M1581 heterologously expressing chloramphenicol biosynthetic gene cluster.

Microbial superhost strains should provide an ideal platform for the efficient homologous or heterologous phenotypic expression of biosynthetic gene clusters (BGCs) of new and novel bioactive molecules. Our aim in the current study was to perform a comparative study at the bioprocess and metabolite levels of the previously designed superhost strain Streptomyces coelicolor M1152 and its derivative strain S. coelicolor M1581 heterologously expressing chloramphenicol BGC. Parent strain M1152 was characterized by a higher specific growth rate, specific CO2 evolution rate, and a higher specific l-glutamate consumption rate as compared with M1581. Intracellular primary central metabolites (nucleoside/sugar phosphates, amino acids, organic acids, and CoAs) were quantified using four targeted LC-MS/MS-based methods. The metabolite pathways in the nonantibiotic producing S. coelicolor host strain were flooded with carbon from both carbon sources, whereas in antibiotic-producing strain, the carbon of l-glutamate seems to be draining out through excreting synthesized antibiotic. The 13 C-isotope-labeling experiments revealed the bidirectionality in the glycolytic pathway and reversibility in the non-oxidative part of PPP even with continuous uptake of d-glucose. The change in the primary metabolites due to the insertion of BGC disclosed a clear linkage between the primary and secondary metabolites.

Large dependency of intracellular NAD and CoA pools on cultivation conditions in Saccharomyces cerevisiae.

OBJECTIVE: The objective of this study was to investigate the variation of NAD and CoA metabolite pools in Saccharomyces cerevisiae cultivated under various cultivation conditions. This study complements a previous report on glycolytic, pentose phosphate pathway, tricarboxylic acid cycle, amino acids, and deoxy-/nucleoside phosphate pools determined under the same cultivation conditions. RESULTS: S. cerevisiae pellets from batch (four carbohydrate sources) and chemostat (carbon-, nitrogen-, phosphate-limited and a range of dilution rates) bioreactor cultivations were extracted and analyzed with two recently established absolute quantitative liquid chromatography mass spectrometry (LC-MS/MS) methods for NAD and CoA metabolites. Both methods apply 13C internal standard dilution strategy for the enhanced analytical accuracy and precision. Individual metabolite pools were relatively constant for the different growth rates within the same mode of cultivation, but large differences were observed among some of the modes, i.e. NAD metabolites were 10 to 100-fold lower in nitrogen limited chemostats compared to the other modes, and phosphate limited chemostats were characterized with much lower CoA metabolite pools. The results complement the previous results and together provide a comprehensive insight into primary metabolite pools variations at a large range in growth and carbon source consumption rates.

Adaptation of central metabolite pools to variations in growth rate and cultivation conditions in Saccharomyces cerevisiae.

BACKGROUND: Saccharomyces cerevisiae is a well-known popular model system for basic biological studies and serves as a host organism for the heterologous production of commercially interesting small molecules and proteins. The central metabolism is at the core to provide building blocks and energy to support growth and survival in normal situations as well as during exogenous stresses and forced heterologous protein production. Here, we present a comprehensive study of intracellular central metabolite pool profiling when growing S. cerevisiae on different carbon sources in batch cultivations and at different growth rates in nutrient-limited glucose chemostats. The latest versions of absolute quantitative mass spectrometry-based metabolite profiling methodology were applied to cover glycolytic and pentose phosphate pathway metabolites, tricarboxylic acid cycle (TCA), complete amino acid, and deoxy-/nucleoside phosphate pools. RESULTS: Glutamate, glutamine, alanine, and citrate were the four most abundant metabolites for most conditions tested. The amino acid is the dominant metabolite class even though a marked relative reduction compared to the other metabolite classes was observed for nitrogen and phosphate limited chemostats. Interestingly, glycolytic and pentose phosphate pathway (PPP) metabolites display the largest variation among the cultivation conditions while the nucleoside phosphate pools are more stable and vary within a closer concentration window. The overall trends for glucose and nitrogen-limited chemostats were increased metabolite pools with the increasing growth rate. Next, comparing the chosen chemostat reference growth rate (0.12 h-1, approximate one-fourth of maximal unlimited growth rate) illuminates an interesting pattern: almost all pools are lower in nitrogen and phosphate limited conditions compared to glucose limitation, except for the TCA metabolites citrate, isocitrate and α-ketoglutarate. CONCLUSIONS: This study provides new knowledge-how the central metabolism is adapting to various cultivations conditions and growth rates which is essential for expanding our understanding of cellular metabolism and the development of improved phenotypes in metabolic engineering.

Absolute Quantification of the Central Carbon Metabolome in Eight Commonly Applied Prokaryotic and Eukaryotic Model Systems.

Absolute quantification of intracellular metabolite pools is a prerequisite for modeling and in-depth biological interpretation of metabolomics data. It is the final step of an elaborate metabolomics workflow, with challenges associated with all steps-from sampling to quantifying the physicochemically diverse metabolite pool. Chromatographic separation combined with mass spectrometric (MS) detection is the superior platform for high coverage, selective, and sensitive detection of metabolites. Herein, we apply our quantitative MS-metabolomics workflow to measure and present the central carbon metabolome of a panel of commonly applied biological model systems. The workflow includes three chromatographic methods combined with isotope dilution tandem mass spectrometry to allow for absolute quantification of 68 metabolites of glycolysis, the pentose phosphate pathway, the tricarboxylic acid cycle, and the amino acid and (deoxy) nucleoside pools. The biological model systems; Bacillus subtilis, Saccharomyces cerevisiae, two microalgal species, and four human cell lines were all cultured in commonly applied culture media and sampled in exponential growth phase. Both literature and databases are scarce with comprehensive metabolite datasets, and existing entries range over several orders of magnitude. The workflow and metabolite panel presented herein can be employed to expand the list of reference metabolomes, as encouraged by the metabolomics community, in a continued effort to develop and refine high-quality quantitative metabolomics workflows.

Cell growth kinetics of Chlorella sorokiniana and nutritional values of its biomass.

The present study investigates the effects of different physico-chemical parameters for the growth of Chlorella sorokiniana and subsequently determination of nutritional values of its biomass. Most suitable temperature, light intensity, pH, and acetic acid concentration were 30°C, 100 µmol m(-2)s(-1), pH 7.5, and 34.8mM, respectively for the growth of this microorganism. Arrhenius growth activation energy, Ea was calculated as 7.08 kJ mol(-1). Monod kinetics constants: maximum specific growth rate (µ max) and substrate (acetic acid) affinity coefficient (Ks) were determined as 0.1 ± 0.01 h(-1) and 76 ± 8 mg L(-1), respectively. Stoichiometric analysis revealed the capture of 1.83 g CO2 and release of 1.9 g O2 for 1g algal biomass synthesis. Algal biomass of C. sorokiniana was found rich in protein and several important minerals such as Mg, Ca, and Fe. Astaxanthin and ß-carotene were extracted and quantified using high performance liquid chromatography.

Physicochemical parameters optimization, and purification of phycobiliproteins from the isolated Nostoc sp.

The present study investigated the effects of several physicochemical parameters on the improvement of phycobiliproteins (especially phycocyanin) synthesis in a newly isolated species of Nostoc sp. Standard BG110 medium was modified to enhance the biomass productivity in different photobioreactors. The initial pH of 8, light intensity of 40 µmol m(-2)s(-1), temperature of 35 °C, diurnal cycle of 16:8 h (light:dark regime), 75.48 µM Na2CO3 and 17.65 mM NaNO3 were found most suitable for the phycobiliproteins synthesis. Cyanobacteria exhibited chromatic adaptation, causing overexpression of phycocyanin in red and phycoerythrin in green light. The maximum phycobiliproteins yield of 0.13 gg(-1) dry cell weight was obtained in green light. Phycocyanin was further purified using thin layer chromatography (TLC), anion exchange chromatography and SDS-PAGE (denaturing gel) electrophoresis.

Carbon dioxide sequestration from industrial flue gas by Chlorella sorokiniana.

The present study investigated the feasibility of using Chlorella sorokiniana for CO2 sequestration from industrial flue gas. The flue gas emitted from the oil producing industry contains mostly CO2 and H2S (15.6% (v/v) and 120 mg L(-1), respectively) along with nitrogen, methane, and other hydrocarbons. The high concentration of CO2 and H2S had an inhibitory effect on the growth of C. sorokiniana. Some efforts were made for the maximization of the algal biomass production using different techniques such as diluted flue gas, flue gas after passing through the scrubber, flue gas passing through serially connected photobioreactors and two different reactors. The highest reduction in the CO2 content of inlet flue gas was 4.1% (v/v). Some new pigments were observed in the flue gas sequestered biomass. Fatty acid composition in the total lipid was determined to evaluate its suitability for food, feed, and biofuel.

Use of image analysis tool for the development of light distribution pattern inside the photobioreactor for the algal cultivation.

Light is one of the important parameters for the growth of photosynthetic microorganisms. In algal photobioreactors, pigmentation of algal cells has additional shading effect which reduces light penetration. Information on the local light intensity inside the photobioreactor is helpful for its efficient designs. Image analysis is based on trichromatic theory and it is used as a tool in studying the light distribution. Digital images of the top view of the photobioreactor were taken and processed using image processing tool in the MATLAB software. This was used to estimate the light intensity distribution in the externally radiating stirred tank photobioreactor across the radial path length. In addition, the effect of light tubes arrangement was studied. This was to find out the effect of light distribution along the periphery of culture suspension. Modified Beer-Lambert's law was found to fit the generated light intensity profile at various cell concentrations and light intensity.

Continuous mode of carbon dioxide sequestration by C. sorokiniana and subsequent use of its biomass for hydrogen production by E. cloacae IIT-BT 08.

The present study investigated to find out the suitability of the CO2 sequestered algal biomass of Chlorella sorokiniana as substrate for the hydrogen production by Enterobacter cloacae IIT-BT 08. The maximum biomass productivity in continuous mode of operation in autotrophic condition was enhanced from 0.05 g L(-1) h(-1) in air to 0.11 g L(-1) h(-1) in 5% air-CO2 (v/v) gas mixture at an optimum dilution rate of 0.05 h(-1). Decrease in steady state biomass and productivity was less sensitive at higher dilution and found fitting with the model proposed by Eppley and Dyer (1965). Pretreated algal biomass of 10 g L(-1) with 2% (v/v) HCl-heat was found most suitable for hydrogen production yielding 9±2 mol H2 (kg COD reduced)(-1) and was found fitting with modified Gompertz equation. Further, hydrogen energy recovery in dark fermentation was significantly enhanced compared to earlier report of hydrogen production by biophotolysis of algae.

Growth characteristics of Chlorella sorokiniana in airlift and bubble column photobioreactors.

The present study investigated the feasibility of bioCO(2) sequestration using Chlorella sorokiniana. It was found that 5% CO(2) (v/v) in air was the most suitable concentration for the growth of this organism. At this concentration, the maximum rate of CO(2) sequestered and the biomass obtained were found to be 1.21 g L(-1)d(-1) and 4.4 g L(-1) respectively. Modeling and simulation of the growth profile was obtained using the logistic equation. Further, at higher CO(2) concentrations, pH drop in the growth media, TAP [-acetate], was prevented by replacing NH(4)Cl by NaNO(3.) Additionally, the study evaluated the performance of two reactors namely: bubble column and airlift reactor based on their growth profile and transport properties like K(L)a and mixing time. The growth profile was better in airlift reactor and it provides cyclic axial mixing of media. K(L)a of downcomer was significantly lower than the riser in airlift reactor.

Development of suitable photobioreactors for CO2 sequestration addressing global warming using green algae and cyanobacteria.

CO(2) sequestration by cyanobacteria and green algae are receiving increased attention in alleviating the impact of increasing CO(2) in the atmosphere. They, in addition to CO(2) capture, can produce renewable energy carriers such as carbon free energy hydrogen, bioethanol, biodiesel and other valuable biomolecules. Biological fixation of CO(2) are greatly affected by the characteristics of the microbial strains, their tolerance to temperature and the CO(2) present in the flue gas including SO(X), NO(X). However, there are additional factors like the availability of light, pH, O(2) removal, suitable design of the photobioreactor, culture density and the proper agitation of the reactor that will affect significantly the CO(2) sequestration process. Present paper deals with the photobioreactors of different geometry available for biomass production. It also focuses on the hybrid types of reactors (integrating two reactors) which can be used for overcoming the bottlenecks of a single photobioreactor.