Melatonin involves hydrogen sulfide in the regulation of H+-ATPase activity, nitrogen metabolism, and ascorbate-glutathione system under chromium toxicity.

Contamination of soils with chromium (Cr) jeopardized agriculture production globally. The current study was planned with the aim to better comprehend how melatonin (Mel) and hydrogen sulfide (H2S) regulate antioxidant defense system, potassium (K) homeostasis, and nitrogen (N) metabolism in tomato seedlings under Cr toxicity. The data reveal that application of 30 µM Mel to the seedlings treated with 25 µM Cr has a positive effect on H2S metabolism that resulted in a considerable increase in H2S. Exogenous Mel improved phytochelatins content and H+-ATPase activity with an associated increase in K content as well. Use of tetraethylammonium chloride (K+-channel blocker) and sodium orthovanadate (H+-ATPase inhibitor) showed that Mel maintained K homeostasis through regulating H+-ATPase activity under Cr toxicity. Supplementation of the stressed seedlings with Mel substantially scavenged excess reactive oxygen species (ROS) that maintained ROS homeostasis. Reduced electrolyte leakage and lipid peroxidation were additional signs of Mel's ROS scavenging effects. In addition, Mel also maintained normal functioning of nitrogen (N) metabolism and ascorbate-glutathione (AsA-GSH) system. Improved level of N fulfilled its requirement for various enzymes that have induced resilience during Cr stress. Additionally, the AsA-GSH cycle's proper operation maintained redox equilibrium, which is necessary for the biological system to function normally. Conversely, 1 mM hypotaurine (H2S scavenger) abolished the Mel-effect and again Cr-induced impairment on the above-mentioned parameters was observed even in presence of Mel. Therefore, based on the observed findings, we concluded that Mel needs endogenous H2S to alleviate Cr-induced impairments in tomato seedlings.

Assessments of some trace metals in water samples of nursery pond of Grass Carp (Ctenopharyngodon idella, Valenciennes, 1844) in Bannu Fish Hatchery of Khyber Pakhtunkhwa, Pakistan / Avaliações de alguns vestígios de metais em amostras de água de viveiro de carpa-capim (Ctenopharyngodon idella, Valenciennes, 1844) em Bannu Fish Hatchery de Khyber Pakhtunkhwa, Paquistão

Abstract The present investigation was aimed to examine the concentrations of trace metals including e.g copper (Cu), manganese (Mn), nickel (Ni), and zinc (Zn) in water samples collected from nursery pond of grass carp (Ctenopharyngodon idella) in Bannu Hatchery of Khyber Pakhtunkhwa during the period from April 2018 to January 2019. The temperature and pH of each water sample were measured for the whole study duration. The concentration of Copper (Cu), Manganese (Mn), Nickel (Ni), and Zinc (Zn) in collected water samples were measured in mg/liter by using Atomic Absorption Spectrophotometer. The blank and standard solutions for device calibration Standard solutions i.e., 2.0 mg, 4.0 mg, and 6.0 were used to measure the concentration of these metals in water samples to verify the measurements. The data was statistically analyzed on descriptive statistics (estimation of proportions and standard deviation) used to summarize mean concentration. The results obtained of both temperature and pH of water samples were found in ranged 10 to 36 0C and 7.0 to 8.44; whereas the size of fry stages was ranged from 4.0 to 56.0 mm in total length. The results of investigated metals found in pond water samples are in order of Zn>Mn>Ni>Cu, respectively. As optimum temperature and pH for grass carp were mostly between 15 0C and 30 0C and pH 6.5 to 8.0. It was concluded from obtained results that temperature, pH, and trace metals were found appropriate for the growth of Ctenopharyngodon idella from fry to fingerling stages, but the highest amount of zinc can cause its mortality. It is a preliminary study on grass carp culturing in Bannu fish hatchery so, it would provide useful information for model fish seed production unit in a hatchery.

Resumo A presente investigação teve como objetivo examinar as concentrações de metais traço, incluindo cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) em amostras de água coletadas de viveiro de carpa-capim (Ctenopharyngodon idella) em Incubatório de Bannu de Khyber Pakhtunkhwa durante o período de abril de 2018 a janeiro de 2019. A temperatura e o pH de cada amostra de água foram medidos durante todo o período do estudo. A concentração de cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) nas amostras de água coletadas foi medida em mg / litro usando espectrofotômetro de absorção atômica. As soluções em branco e padrão para as soluções padrão de calibração do dispositivo, ou seja, 2,0 mg, 4,0 mg e 6,0, foram usadas para medir a concentração desses metais em amostras de água para verificar as medições. Os dados foram analisados ​​estatisticamente em estatística descritiva (estimativa de proporções e desvio padrão) usada para resumir a concentração média. Os resultados obtidos tanto de temperatura quanto de pH das amostras de água variaram de 10 a 36 0C e 7,0 a 8,44, enquanto o tamanho dos estágios de alevinos variou de 4,0 a 56,0 mm no comprimento total. Os resultados dos metais investigados encontrados em amostras de água de lagoas são da ordem de Zn > Mn > Ni > Cu, respectivamente. A temperatura e o pH ótimos para a carpa-capim foram principalmente entre 15 0C e 30 0C e pH 6,5 a 8,0. Concluiu-se, a partir dos resultados obtidos, que temperatura, pH e traços de metais foram considerados adequados para o crescimento de Ctenopharyngodon idella desde os estágios de alevinos até alevinos, mas a maior quantidade de zinco pode causar sua mortalidade. É um estudo preliminar sobre a cultura da carpa-capim em incubatório de peixes Bannu, portanto forneceria informações úteis para modelo de unidade de produção de sementes de peixes em um incubatório.

Assessments of some trace metals in water samples of nursery pond of Grass Carp (Ctenopharyngodon idella, Valenciennes, 1844) in Bannu Fish Hatchery of Khyber Pakhtunkhwa, Pakistan / Avaliações de alguns vestígios de metais em amostras de água de viveiro de carpa capim (Ctenopharyngodon idella, Valenciennes, 1844) em Bannu Fish Hatchery de Khyber Pakhtunkhwa, Paquistão

The present investigation was aimed to examine the concentrations of trace metals including e.g copper (Cu), manganese (Mn), nickel (Ni), and zinc (Zn) in water samples collected from nursery pond of grass carp (Ctenopharyngodon idella) in Bannu Hatchery of Khyber Pakhtunkhwa during the period from April 2018 to January 2019. The temperature and pH of each water sample were measured for the whole study duration. The concentration of Copper (Cu), Manganese (Mn), Nickel (Ni), and Zinc (Zn) in collected water samples were measured in mg/liter by using Atomic Absorption Spectrophotometer. The blank and standard solutions for device calibration Standard solutions i.e., 2.0 mg, 4.0 mg, and 6.0 were used to measure the concentration of these metals in water samples to verify the measurements. The data was statistically analyzed on descriptive statistics (estimation of proportions and standard deviation) used to summarize mean concentration. The results obtained of both temperature and pH of water samples were found in ranged 10 to 36 0C and 7.0 to 8.44; whereas the size of fry stages was ranged from 4.0 to 56.0 mm in total length. The results of investigated metals found in pond water samples are in order of Zn>Mn>Ni>Cu, respectively. As optimum temperature and pH for grass carp were mostly between 15 0C and 30 0C and pH 6.5 to 8.0. It was concluded from obtained results that temperature, pH, and trace metals were found appropriate for the growth of Ctenopharyngodon idella from fry to fingerling stages, but the highest amount of zinc can cause its mortality. It is a preliminary study on grass carp culturing in Bannu fish hatchery so, it would provide useful information for model fish seed production unit in a hatchery.

A presente investigação teve como objetivo examinar as concentrações de metais traço, incluindo cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) em amostras de água coletadas de viveiro de carpa-capim (Ctenopharyngodon idella) em Incubatório de Bannu de Khyber Pakhtunkhwa durante o período de abril de 2018 a janeiro de 2019. A temperatura e o pH de cada amostra de água foram medidos durante todo o período do estudo. A concentração de cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) nas amostras de água coletadas foi medida em mg / litro usando espectrofotômetro de absorção atômica. As soluções em branco e padrão para as soluções padrão de calibração do dispositivo, ou seja, 2,0 mg, 4,0 mg e 6,0, foram usadas para medir a concentração desses metais em amostras de água para verificar as medições. Os dados foram analisados estatisticamente em estatística descritiva (estimativa de proporções e desvio padrão) usada para resumir a concentração média. Os resultados obtidos tanto de temperatura quanto de pH das amostras de água variaram de 10 a 36 0C e 7,0 a 8,44, enquanto o tamanho dos estágios de alevinos variou de 4,0 a 56,0 mm no comprimento total. Os resultados dos metais investigados encontrados em amostras de água de lagoas são da ordem de Zn > Mn > Ni > Cu, respectivamente. A temperatura e o pH ótimos para a carpa-capim foram principalmente entre 15 0C e 30 0C e pH 6,5 a 8,0. Concluiu-se, a partir dos resultados obtidos, que temperatura, pH e traços de metais foram considerados adequados para o crescimento de Ctenopharyngodon idella desde os estágios de alevinos até alevinos, mas a maior quantidade de zinco pode causar sua mortalidade. É um estudo preliminar sobre a cultura da carpa-capim em incubatório de peixes Bannu, portanto forneceria informações úteis para modelo de unidade de produção de sementes de peixes em um incubatório.

Assessments of some trace metals in water samples of nursery pond of Grass Carp (Ctenopharyngodon idella, Valenciennes, 1844) in Bannu Fish Hatchery of Khyber Pakhtunkhwa, Pakistan

Abstract The present investigation was aimed to examine the concentrations of trace metals including e.g copper (Cu), manganese (Mn), nickel (Ni), and zinc (Zn) in water samples collected from nursery pond of grass carp (Ctenopharyngodon idella) in Bannu Hatchery of Khyber Pakhtunkhwa during the period from April 2018 to January 2019. The temperature and pH of each water sample were measured for the whole study duration. The concentration of Copper (Cu), Manganese (Mn), Nickel (Ni), and Zinc (Zn) in collected water samples were measured in mg/liter by using Atomic Absorption Spectrophotometer. The blank and standard solutions for device calibration Standard solutions i.e., 2.0 mg, 4.0 mg, and 6.0 were used to measure the concentration of these metals in water samples to verify the measurements. The data was statistically analyzed on descriptive statistics (estimation of proportions and standard deviation) used to summarize mean concentration. The results obtained of both temperature and pH of water samples were found in ranged 10 to 36 0C and 7.0 to 8.44; whereas the size of fry stages was ranged from 4.0 to 56.0 mm in total length. The results of investigated metals found in pond water samples are in order of Zn>Mn>Ni>Cu, respectively. As optimum temperature and pH for grass carp were mostly between 15 0C and 30 0C and pH 6.5 to 8.0. It was concluded from obtained results that temperature, pH, and trace metals were found appropriate for the growth of Ctenopharyngodon idella from fry to fingerling stages, but the highest amount of zinc can cause its mortality. It is a preliminary study on grass carp culturing in Bannu fish hatchery so, it would provide useful information for model fish seed production unit in a hatchery.

Resumo A presente investigação teve como objetivo examinar as concentrações de metais traço, incluindo cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) em amostras de água coletadas de viveiro de carpa-capim (Ctenopharyngodon idella) em Incubatório de Bannu de Khyber Pakhtunkhwa durante o período de abril de 2018 a janeiro de 2019. A temperatura e o pH de cada amostra de água foram medidos durante todo o período do estudo. A concentração de cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) nas amostras de água coletadas foi medida em mg / litro usando espectrofotômetro de absorção atômica. As soluções em branco e padrão para as soluções padrão de calibração do dispositivo, ou seja, 2,0 mg, 4,0 mg e 6,0, foram usadas para medir a concentração desses metais em amostras de água para verificar as medições. Os dados foram analisados estatisticamente em estatística descritiva (estimativa de proporções e desvio padrão) usada para resumir a concentração média. Os resultados obtidos tanto de temperatura quanto de pH das amostras de água variaram de 10 a 36 0C e 7,0 a 8,44, enquanto o tamanho dos estágios de alevinos variou de 4,0 a 56,0 mm no comprimento total. Os resultados dos metais investigados encontrados em amostras de água de lagoas são da ordem de Zn > Mn > Ni > Cu, respectivamente. A temperatura e o pH ótimos para a carpa-capim foram principalmente entre 15 0C e 30 0C e pH 6,5 a 8,0. Concluiu-se, a partir dos resultados obtidos, que temperatura, pH e traços de metais foram considerados adequados para o crescimento de Ctenopharyngodon idella desde os estágios de alevinos até alevinos, mas a maior quantidade de zinco pode causar sua mortalidade. É um estudo preliminar sobre a cultura da carpa-capim em incubatório de peixes Bannu, portanto forneceria informações úteis para modelo de unidade de produção de sementes de peixes em um incubatório.

Monitoring pond water quality to improve the production of Labeo rohita (Hamilton, 1822) in Bannu Fish Hatchery of Bannu district, Khyber Pakhtunkhwa province; An Implications for artificial fish culture

Abstract The water quality parameters of a fish pond are essential to be managing properly under control for successful operations of fish culture. Improper management of pond water quality during the juvenile stages can create stressful conditions to produce various harmful diseases, which may decrease the fish quality and results in low profits. The present study was aimed to monitored important water quality parameters of nursery ponds of Labeo rohita culturing in Bannu fish hatchery. The study duration was 75 days extends from 10th June to 24th August 2019 for the successful culture of this specie which can play a significant role in breeding season. Furthermore, the concentration of some heavy metals like copper (Cu), nickel (Ni), manganese (Mn), iron (Fe), cadmium (Cd), and zinc (Zn) in pond water and fry stages of this species was also determined. The data obtained from all water quality parameters were analyzed expressed as range, mean and standard deviation using MS Excel 2013. The obtained results of 75 days study revealed that the water pH & temperature, electric conductivity, total dissolved solids (TDS), and total dissolved oxygen (DO) of pond water samples were found within a tolerable limit except salinity and dissolved ammonia concentration were not permissible for fish growth. The average concentration of heavy metals in pond water exhibited descending order Fe>Ni>Mn>Zn>Cd>Ni, which was found in acceptable ranges. Whereas, the average values of heavy metals in fry stages were in the order of Zn>Fe>Ni>Cu>Cd>Mn, and found within the recommended values of WHO/FAO. Thus, it was concluded from this study that good water quality is a precondition, maintaining balanced levels of water quality parameters is fundamental for both the health and growth of fish culture which is quite necessary for assuring increased fish productivity. It is recommended to monitor and assess water quality parameters on a routine basis for promoting healthy fish culture.

Resumo Os parâmetros de qualidade da água de um viveiro de peixes são essenciais para um manejo adequado e sob controle para operações bem-sucedidas de piscicultura. O manejo inadequado da qualidade da água do tanque durante os estágios juvenis pode criar condições estressantes para a produção de várias doenças nocivas, o que pode diminuir a qualidade do peixe e resultar em baixos lucros. O presente estudo teve como objetivo monitorar importantes parâmetros de qualidade da água de viveiros de cultivo de Labeo rohita em incubadora de peixes Bannu. A duração do estudo foi de 75 dias, estendendo-se de 10 de junho a 24 de agosto de 2019 para o sucesso do cultivo dessa espécie que pode desempenhar papel significativo na época de reprodução. Além disso, a concentração de alguns metais pesados como cobre (Cu), níquel (Ni), manganês (Mn), ferro (Fe), cádmio (Cd) e zinco (Zn) na água do tanque e estágios de fritura dessa espécie também foram determinados. Os dados obtidos de todos os parâmetros de qualidade da água foram analisados expressos como faixa, média e desvio padrão usando o MS Excel 2013. Os resultados obtidos de 75 dias de estudo revelaram que o pH e temperatura da água, condutividade elétrica, sólidos totais dissolvidos (TDS) e total de oxigênio dissolvido (OD) das amostras de água do lago foram encontrados dentro de um limite tolerável, exceto salinidade e concentração de amônia dissolvida não eram permitidas para o crescimento dos peixes. A concentração média de metais pesados na água da lagoa apresentou ordem decrescente Fe > Ni > Mn > Zn > Cd > Ni, que foi encontrada em faixas aceitáveis. Já os valores médios dos metais pesados nos estágios de fritura foram da ordem de Zn > Fe > Ni > Cu > Cd > Mn, e encontrados dentro dos valores recomendados pela OMS/FAO. Assim, concluiu-se deste estudo que a boa qualidade da água é uma condição prévia, manter níveis equilibrados dos parâmetros de qualidade da água é fundamental para a saúde e crescimento da piscicultura, o que é bastante necessário para garantir o aumento da produtividade piscícola. Recomendam-se monitorar e avaliar os parâmetros de qualidade da água em uma base rotineira para promover a piscicultura saudável.

Ploidy Status, Nuclear DNA Content and Start Codon Targeted (SCoT) Genetic Homogeneity Assessment in Digitalis purpurea L., Regenerated In Vitro.

Digitalis purpurea L. is a therapeutically important plant that synthesizes important cardiotonics such as digitoxin and digoxin. The present work reports a detailed and efficient propagation protocol for D. purpurea by optimizing various PGR concentrations in Murashige and Skoog (MS) medium. The genetic homogeneity of in vitro regenerants was assessed by the flow cytometric method (FCM) and Start Codon Targeted (SCoT) marker technique. Firstly, the seeds inoculated in full MS medium added with 0.5 mg/L GA3 produced seedlings. Different parts such as hypocotyl, nodes, leaves and apical shoots were used as explants. The compact calli were obtained on BAP alone or in combinations with 2, 4-D/NAA. The hypocotyl-derived callus induced somatic embryos which proliferated and germinated best in 0.75 mg/L BAP-fortified MS medium. Scanning electron microscopic (SEM) images confirmed the presence of various developmental stages of somatic embryos. Shoot regeneration was obtained in which BAP at 1.0 mg/L and 2.0 mg/L BAP + 0.5 mg/L 2,4-D proved to be the best treatments of PGRs in inducing direct and indirect shoot buds. The regenerated shoots showed the highest rooting percentage (87.5%) with 24.7 ± 1.9 numbers of roots/shoot in 1.0 mg/L IBA augmented medium. The rooted plantlets were acclimatized in a greenhouse at a survival rate of 85-90%. The genome size and the 2C nuclear DNA content of field-grown, somatic embryo-regenerated and organogenic-derived plants were estimated and noted to be 3.1, 3.2 and 3.0 picogram (pg), respectively; there is no alteration in ploidy status and the DNA content, validating genetic uniformity. Six SCoT primers unveiled 94.3%-95.13% monomorphic bands across all the plant samples analyzed, further indicating genetic stability among in vitro clones and mother plants. This study describes for the first time successful induction of somatic embryos from hypocotyl callus; and flow cytometry and SCoT marker confirmed the genetic homogeneity of regenerated plants.

T cell receptor sequencing identifies prior SARS-CoV-2 infection and correlates with neutralizing antibodies and disease severity.

BACKGROUNDMeasuring the immune response to SARS-CoV-2 enables assessment of past infection and protective immunity. SARS-CoV-2 infection induces humoral and T cell responses, but these responses vary with disease severity and individual characteristics.METHODSA T cell receptor (TCR) immunosequencing assay was conducted using small-volume blood samples from 302 individuals recovered from COVID-19. Correlations between the magnitude of the T cell response and neutralizing antibody (nAb) titers or indicators of disease severity were evaluated. Sensitivity of T cell testing was assessed and compared with serologic testing.RESULTSSARS-CoV-2-specific T cell responses were significantly correlated with nAb titers and clinical indicators of disease severity, including hospitalization, fever, and difficulty breathing. Despite modest declines in depth and breadth of T cell responses during convalescence, high sensitivity was observed until at least 6 months after infection, with overall sensitivity ~5% greater than serology tests for identifying prior SARS-CoV-2 infection. Improved performance of T cell testing was most apparent in recovered, nonhospitalized individuals sampled > 150 days after initial illness, suggesting greater sensitivity than serology at later time points and in individuals with less severe disease. T cell testing identified SARS-CoV-2 infection in 68% (55 of 81) of samples with undetectable nAb titers (<1:40) and in 37% (13 of 35) of samples classified as negative by 3 antibody assays.CONCLUSIONThese results support TCR-based testing as a scalable, reliable measure of past SARS-CoV-2 infection with clinical value beyond serology.TRIAL REGISTRATIONSpecimens were accrued under trial NCT04338360 accessible at clinicaltrials.gov.FUNDINGThis work was funded by Adaptive Biotechnologies, Frederick National Laboratory for Cancer Research, NIAID, Fred Hutchinson Joel Meyers Endowment, Fast Grants, and American Society for Transplantation and Cell Therapy.

Longitudinal analysis of T cell receptor repertoires reveals shared patterns of antigen-specific response to SARS-CoV-2 infection.

T cells play a prominent role in orchestrating the immune response to viral diseases, but their role in the clinical presentation and subsequent immunity to SARS-CoV-2 infection remains poorly understood. As part of a population-based survey of the municipality of Vo', Italy, conducted after the initial SARS-CoV-2 outbreak, we sampled the T cell receptor (TCR) repertoires of the population 2 months after the initial PCR survey and followed up positive cases 9 and 15 months later. At 2 months, we found that 97.0% (98 of 101) of cases had elevated levels of TCRs associated with SARS-CoV-2. T cell frequency (depth) was increased in individuals with more severe disease. Both depth and diversity (breadth) of the TCR repertoire were positively associated with neutralizing antibody titers, driven mostly by CD4+ T cells directed against spike protein. At the later time points, detection of these TCRs remained high, with 90.7% (78 of 96) and 86.2% (25 of 29) of individuals having detectable signal at 9 and 15 months, respectively. Forty-three individuals were vaccinated by month 15 and showed a significant increase in TCRs directed against spike protein. Taken together, these results demonstrate the central role of T cells in mounting an immune defense against SARS-CoV-2 that persists out to 15 months.

Monitoring pond water quality to improve the production of Labeo rohita (Hamilton, 1822) in Bannu Fish Hatchery of Bannu district, Khyber Pakhtunkhwa province; An Implications for artificial fish culture.

The water quality parameters of a fish pond are essential to be managing properly under control for successful operations of fish culture. Improper management of pond water quality during the juvenile stages can create stressful conditions to produce various harmful diseases, which may decrease the fish quality and results in low profits. The present study was aimed to monitored important water quality parameters of nursery ponds of Labeo rohita culturing in Bannu fish hatchery. The study duration was 75 days extends from 10th June to 24th August 2019 for the successful culture of this specie which can play a significant role in breeding season. Furthermore, the concentration of some heavy metals like copper (Cu), nickel (Ni), manganese (Mn), iron (Fe), cadmium (Cd), and zinc (Zn) in pond water and fry stages of this species was also determined. The data obtained from all water quality parameters were analyzed expressed as range, mean and standard deviation using MS Excel 2013. The obtained results of 75 days study revealed that the water pH & temperature, electric conductivity, total dissolved solids (TDS), and total dissolved oxygen (DO) of pond water samples were found within a tolerable limit except salinity and dissolved ammonia concentration were not permissible for fish growth. The average concentration of heavy metals in pond water exhibited descending order Fe>Ni>Mn>Zn>Cd>Ni, which was found in acceptable ranges. Whereas, the average values of heavy metals in fry stages were in the order of Zn>Fe>Ni>Cu>Cd>Mn, and found within the recommended values of WHO/FAO. Thus, it was concluded from this study that good water quality is a precondition, maintaining balanced levels of water quality parameters is fundamental for both the health and growth of fish culture which is quite necessary for assuring increased fish productivity. It is recommended to monitor and assess water quality parameters on a routine basis for promoting healthy fish culture.

Assessments of some trace metals in water samples of nursery pond of Grass Carp (Ctenopharyngodon idella, Valenciennes, 1844) in Bannu Fish Hatchery of Khyber Pakhtunkhwa, Pakistan.

The present investigation was aimed to examine the concentrations of trace metals including e.g copper (Cu), manganese (Mn), nickel (Ni), and zinc (Zn) in water samples collected from nursery pond of grass carp (Ctenopharyngodon idella) in Bannu Hatchery of Khyber Pakhtunkhwa during the period from April 2018 to January 2019. The temperature and pH of each water sample were measured for the whole study duration. The concentration of Copper (Cu), Manganese (Mn), Nickel (Ni), and Zinc (Zn) in collected water samples were measured in mg/liter by using Atomic Absorption Spectrophotometer. The blank and standard solutions for device calibration Standard solutions i.e., 2.0 mg, 4.0 mg, and 6.0 were used to measure the concentration of these metals in water samples to verify the measurements. The data was statistically analyzed on descriptive statistics (estimation of proportions and standard deviation) used to summarize mean concentration. The results obtained of both temperature and pH of water samples were found in ranged 10 to 36 0C and 7.0 to 8.44; whereas the size of fry stages was ranged from 4.0 to 56.0 mm in total length. The results of investigated metals found in pond water samples are in order of Zn>Mn>Ni>Cu, respectively. As optimum temperature and pH for grass carp were mostly between 15 0C and 30 0C and pH 6.5 to 8.0. It was concluded from obtained results that temperature, pH, and trace metals were found appropriate for the growth of Ctenopharyngodon idella from fry to fingerling stages, but the highest amount of zinc can cause its mortality. It is a preliminary study on grass carp culturing in Bannu fish hatchery so, it would provide useful information for model fish seed production unit in a hatchery.

Development and validation of a reversed-phase HPLC method for quantification of 1'-acetoxychavicol acetate content in a nanostructured lipid carrier formulation

Abstract 1'-acetoxychavicol acetate (ACA)-loaded nanostructured lipid carriers (NLCs) were formulated for prostate cancer therapy and to determine the optimal therapeutic dose, we developed a rapid, specific, and accurate reversed-phase high-performance liquid chromatography (RP-HPLC) method to quantify the ACA content in NLCs. The method was validated according to International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines. Chromatographic separation of ACA from the lipid components was performed with an Agilent 1220 Infinity LC system and ultraviolet detector using an Agilent Poroshell C18 column (4.6 x 250.0 mm). The mobile phase consisted of acetonitrile and water (80:20 [v/v]) with a flow rate of 0.8 mL/min in isocratic mode. Linearity of the standard curve was assessed at an ACA concentration range of 5-200 µg/mL, and a 1/x weighted linear regression was adopted for the calibration curve. The calculated limits of detection and quantification were 0.59 µg/mL and 1.79 µg/mL, respectively. The mean percent recovery of ACA was 100.02% (relative SD, 2%), and the coefficients of variation for intraday and interday assays were within the values required by the ICH. We also demonstrated robustness of the method by altering the mobile phase ratio and flow rate. Furthermore, we proved specificity of the method for ACA by comparing chromatograms of the blank NLC and ACA-NLC. Hence, we effectively used this validated method to determine the drug-loading capacity and entrapment efficiency of the NLCs.

Chlorine taste can increase simulated exposure to both fecal contamination and disinfection byproducts in water supplies.

Expanding drinking water chlorination could substantially reduce the burden of disease in low- and middle-income countries, but the taste of chlorinated water often impedes adoption. We developed a Monte Carlo simulation to estimate the effect of people's choice to accept or reject drinking water based on chlorine taste and their subsequent exposure to E. coli and trihalomethanes, a class of disinfection byproduct (DBP). The simulation used empirical data from Dhaka, Bangladesh, a megacity with endemic waterborne disease. We drew on published taste acceptability thresholds from Dhaka residents, measured residual chlorine and thermotolerant E. coli inactivation following the addition of six chlorine doses (0.25-3.0 mg/L as Cl2) to untreated piped water samples from 100 locations, and analyzed trihalomethane formation in 54 samples. A dose of 0.5 mg/L, 75% lower than the 2 mg/L dose typically recommended for household chlorination of low-turbidity waters, minimized overall exposure to E. coli. Doses of 1-2 mg/L maximized overall exposure to trihalomethanes. Accounting for chlorine taste aversion indicates that microbiological exposure increases and DBP exposure decreases above certain doses as a higher proportion of people reject chlorinated water in favor of untreated water. Taken together with findings from other modeling analyses, empirical studies, and field trials, our results suggest that taste acceptability should be a critical consideration in establishing chlorination dosing guidelines. Particularly when chlorination is first implemented in water supplies with low chlorine demand, lower doses than those generally recommended for household water treatment can help avoid taste-related objections while still meaningfully reducing contaminant exposure.

The Usages and Potential Uses of Alginate for Healthcare Applications.

Due to their unique properties, alginate-based biomaterials have been extensively used to treat different diseases, and in the regeneration of diverse organs. A lot of research has been done by the different scientific community to develop biofilms for fulfilling the need for sustainable human health. The aim of this review is to hit upon a hydrogel enhancing the scope of utilization in biomedical applications. The presence of active sites in alginate hydrogels can be manipulated for managing various non-communicable diseases by encapsulating, with the bioactive component as a potential site for chemicals in developing drugs, or for delivering macromolecule nutrients. Gels are accepted for cell implantation in tissue regeneration, as they can transfer cells to the intended site. Thus, this review will accelerate advanced research avenues in tissue engineering and the potential of alginate biofilms in the healthcare sector.

A review: Pharmacological aspects of metal based 1,2,4-triazole derived Schiff bases.

Clinical reports have highlighted the radical increase of antibiotic resistance. As a result, multidrug resistance has emerged as a serious threat to human health. Many organic compounds commonly used as drugs in the past, no longer have pure organic mode of action rather need bio-transformation or more activation. Bulk of research has shown that they need trace amount of metal ions incorporated within the chemistry of bioactive molecules for enhancement of their potentiality to fight aggressively against resistance. The deficiency of some metal ions can also be responsible for many diseases like growth retardation, pernicious anemia and heart diseases in infants. To overcome these problems, there is a need to introduce novel strategies which have new mechanism of action along with significant spectrum of biological activity, enhanced safety and efficacy. Bioinorganic compounds have played imperative role in developing the new strategy in the form of "Metal Based Drugs". In current years there have been momentous rise of interest in the application of metal based Schiff base compounds to treat various diseases which are difficult to be treated with conventional methodologies. The unique properties of metal chelates acting as an intermediate between conventional organic and inorganic compounds provided innovative opportunities in the field of pharmaceutical chemistry. In this review, we have exclusively focused on the search of metal based 1,2,4-triazole derived Schiff base compounds (synthesized, reported and reviewed in the past ten years) that possess various biological activities such as antifungal, antibacterial, antioxidant, antidiabetic, anthelmintic, anticancer, antiproliferative, cytotoxic and DNA-intercalation activity.

T-cell receptor sequencing identifies prior SARS-CoV-2 infection and correlates with neutralizing antibody titers and disease severity.

Measuring the adaptive immune response to SARS-CoV-2 can enable the assessment of past infection as well as protective immunity and the risk of reinfection. While neutralizing antibody (nAb) titers are one measure of protection, such assays are challenging to perform at a large scale and the longevity of the SARS-CoV-2 nAb response is not fully understood. Here, we apply a T-cell receptor (TCR) sequencing assay that can be performed on a small volume standard blood sample to assess the adaptive T-cell response to SARS-CoV-2 infection. Samples were collected from a cohort of 302 individuals recovered from COVID-19 up to 6 months after infection. Previously published findings in this cohort showed that two commercially available SARS-CoV-2 serologic assays correlate well with nAb testing. We demonstrate that the magnitude of the SARS-CoV-2-specific T-cell response strongly correlates with nAb titer, as well as clinical indicators of disease severity including hospitalization, fever, or difficulty breathing. While the depth and breadth of the T-cell response declines during convalescence, the T-cell signal remains well above background with high sensitivity up to at least 6 months following initial infection. Compared to serology tests detecting binding antibodies to SARS-CoV-2 spike and nucleoprotein, the overall sensitivity of the TCR-based assay across the entire cohort and all timepoints was approximately 5% greater for identifying prior SARS-CoV-2 infection. Notably, the improved performance of T-cell testing compared to serology was most apparent in recovered individuals who were not hospitalized and were sampled beyond 150 days of their initial illness, suggesting that antibody testing may have reduced sensitivity in individuals who experienced less severe COVID-19 illness and at later timepoints. Finally, T-cell testing was able to identify SARS-CoV-2 infection in 68% (55/81) of convalescent samples having nAb titers below the lower limit of detection, as well as 37% (13/35) of samples testing negative by all three antibody assays. These results demonstrate the utility of a TCR-based assay as a scalable, reliable measure of past SARS-CoV-2 infection across a spectrum of disease severity. Additionally, the TCR repertoire may be useful as a surrogate for protective immunity with additive clinical value beyond serologic or nAb testing methods.

Determining the National Cataract Surgical Rate in Pakistan.

PURPOSE: Cataract surgical rate (CSR) (cataract surgeries performed per million population) is an eye health indicator that helps assess the state of eye care services. A survey in 2002 revealed a CSR of 2254. The current survey aimed to establish a new and sustainable development goal compliant baseline for the volume of cataract surgery performed by different service providers in Pakistan at district, provincial, and national levels. METHODS: The survey was commissioned by the National Committee for Eye Health under the Ministry of National Health Services, Regulations and Coordination. The methodology used for the survey included identification and mapping of all service providers by district and category and data collection from all districts in the country. RESULTS: There were more cataract surgeries performed in women than men with a male-to-female ratio of 0.95. About 98.9% of all cataract surgeries were performed with intraocular lenses, while 63.9% were performed by phacoemulsification. About 17.7% of cataract surgical services were provided in the government sector (including Forces), while nongovernmental organizations and the private sector contributed to 82.3%. Pakistan achieved a national CSR of 5307 which is almost double the CSR determined in 2002. CONCLUSION: In order to achieve a CSR of 7500+ by 2030, there is a need for at least 1,840,000 cataract surgeries to be performed annually. If there is no change in the current annual cataract surgical output, the CSR will drop to 4628 by 2030.

Oncolytic HSV Therapy Modulates Vesicular Trafficking Inducing Cisplatin Sensitivity and Antitumor Immunity.

PURPOSE: Here we investigated the impact of oncolytic herpes simplex virus (HSV) treatment on cisplatin sensitivity of platinum-resistant ovarian cancer, and the impact of the combination on immunotherapy. EXPERIMENTAL DESIGN: Therapeutic efficacy of the combination was assessed in platinum-resistant human and murine ovarian cancer peritoneal metastatic mouse models (n = 9-10/group). RNA sequencing along with flow cytometry of splenocytes from treated mice was employed to examine the effect of antitumor immune response (n = 3/group). Anti-PD-1 antibody was performed to evaluate impact on checkpoint inhibition in vivo. RESULTS: Gene Ontology pathway analysis uncovered disruption of cellular extracellular vesicle (EV)-related pathways in infected cells (FDR = 2.97E-57). Mechanistically, we identified reduced expression of transporters expressed on EV implicated in cisplatin efflux. The increased cisplatin retention led to increased cisplatin-DNA adducts, which resulted in micronuclei and the subsequent activation of cGAS-STING pathway with a significant activation of innate immune cells and translated to an increase in antitumor immunity and efficacy. In mice bearing platinum-resistant ovarian cancer, we also observed a feedback induction of PD-L1 on tumor cells, which sensitized combination-treated mice to anti-PD-1 immune checkpoint therapy. CONCLUSIONS: To our knowledge, this is the first report to show HSV-induced cisplatin retention in infected cells. The consequential increased damaged DNA was then expelled from cells as micronuclei which resulted in induction of inflammatory responses and education of antitumor immunity. The combination therapy also created an environment that sensitized tumors to immune checkpoint therapy.