Arsenic Levels and Seasonal Variation in Pasture Soil, Forage and Horse Blood Plasma in Central Punjab, Pakistan.

The present study aimed to determine the accumulation levels of arsenic in pasture soil, forage and animals. An atomic absorption spectrophotometer was used to determine the concentration of metals in the samples of soil, forage and blood plasma of horses. The level of arsenic ranged between 4.26 mg/kg (summer) and 4.66 mg/kg (winter) in soil samples and 2.67 mg/kg (summer) and 2.94 mg/kg (winter) in forage samples. In blood plasma samples, the mean arsenic (As) values varied between 1.38 and 1.52 mg/L. In the blood plasma samples, the mean As values varied between 1.38 and 1.52 mg/L. No statistically significant changes were observed for arsenic concentrations in plasma samples of horses for sampling seasons (p > 0.05). A positive correlation was observed for forage and blood plasma to a certain degree for arsenic but for other media, arsenic correlations were negative and insignificant. It is therefore suggested that regular monitoring of heavy metals in soils/plants/animals is essential to prevent excessive build-up of arsenic.

Pilot scale production of recombinant hemicellulases and their saccharification potential.

Synergistic saccharification ability of hemicellulases (endo-xylanase and ß-xylosidase) was evaluated in this study for the bioethanol production from plant biomass. Endo-xylanase and ß-xylosidase genes from Bacillus licheniformis were cloned and expressed in Escherichia coli BL21 (DE3). Maximum endo-xylanase production was obtained at 200 rpm agitation speed, air supply rate 2.0 vvm, 70% volume of the medium, 20% dissolved oxygen level and with 3% inoculum size. The optimal conditions for maximum production of recombinant ß-xylosidase enzyme at pilot scale were 200 rpm agitation speed, 25% dissolved oxygen level, 2.5 vvm aeration rate, 70% volume of the medium with 2% inoculum size. Furthermore, the saccharification potential of these recombinant enzymes was checked for the production of xylose sugar by bioconversion of plant biomass by optimizing individually as well as synergistically by optimizing various parameters. Maximum saccharification (93%) of plant biomass was observed when both enzymes were used at a time with 8% sugarcane bagasse as a substrate and 200 units of each enzyme after incubation of 6 hr at 50 °C and 120 rpm. The results obtained in this study suggested these recombinant hemicellulases as potential candidates for the conversion of complex agricultural residues into simple sugars for ultimate use in the biofuel industry.

Determinants of overall knowledge of and attitudes towards HIV/AIDS transmission among ever-married women in Pakistan: evidence from the Demographic and Health Survey 2012-13.

BACKGROUND: HIV/AIDS has emerged as a serious public health issue across the globe, and particularly in developing countries. Comprehensive knowledge and positive attitudes are cornerstones for the prevention, control and treatment of HIV/AIDS. However, there are various misconceptions associated with HIV/AIDS transmission, which lead to negative attitudes towards people living with AIDS. The present study aims to explore the effects of these determinants, related to socio-demographic characteristics and autonomy, on women's overall knowledge and attitudes regarding HIV/AIDS in Pakistan. METHODS: Secondary data analysis was carried out using the national representative dataset of the 2012-13 Pakistan Demographic and Health Survey. A series of questions related to HIV/AIDS was asked of 13,558 ever-married women aged 15-49 years to assess respondents' knowledge regarding modes of HIV/AIDS transmission and preventative measures, as well as their attitudes towards people living with HIV/AIDS. Descriptive and bivariate statistics were used to identify associations with socio-demographic and autonomy-related variables. Furthermore, bivariate and multivariate logistic regression analyses were performed to assess the association between multiple factors and overall HIV/AIDS knowledge as well as attitudes towards people living with AIDS. RESULTS: The results show that only 42% of Pakistani women have heard about HIV/AIDS. Amongst these women, the majority (68%) have good overall knowledge of HIV/AIDS and more than 55% have positive attitudes towards people living with AIDS. Furthermore, women residing in urban areas, having at least secondary-level education, with high autonomy, belonging to the richest wealth quintile and having exposure to mass media had high overall knowledge and positive attitudes towards people living with AIDS. CONCLUSION: The findings of this research support the relevance of women's autonomy, education and exposure to mass media, particularly in rural areas of Pakistan, to address the lack of knowledge and eliminate various myths and stigmatisation of people living with HIV/AIDS. Furthermore, it reveals a need to increase focused and targeted interventions to enhance women's knowledge and positive attitudes towards people living with HIV/AIDS. In this regard, the media can play a proactive role to gauge wider audience in creating awareness and eradicating the myths and misconceptions regarding HIV/AIDS.

Purification and Characterization of a recombinant ß-Xylosidase from Bacillus licheniformis ATCC 14580 into E. coli Bl21.

Present research work is aimed to purify and characterize a recombinant ß-xylosidase enzyme which was previously cloned from Bacillus licheniformis ATCC 14580 in to Escherichia coli BL21. Purification of recombinant enzyme was carried out by using ammonium sulphate precipitation method followed by single step immobilized metal ion affinity chromatography. Specific activity of purified recombinant ß-xylosidase enzyme was 20.78 Umg-1 with 2.58 purification fold and 33.75% recovery. SDS-PAGE was used to determine the molecular weight of recombinant purified ß-xylosidase and it was recorded as 52 kDa. Purified enzyme showed stability upto 90°C within a pH range of 3-8 with and optimal temperature and pH, 55ºC and 7.0, respectively. The enzyme activity was not considerably affected in the presence of EDTA. An increase in the enzyme activity was found in the manifestation of Mg+2. Enzyme activity was also increased by 6%, 18% and 22% in the presence of 1% Tween 80, ß-mercaptoethanol and DTT, respectively. Higher concentrations (10 - 40%) of organic solvents did not show any effect upon activity of enzyme. All these characteristics of the recombinant enzyme endorsed it as a potential candidate for biofuel industry.

Purification and characterization of cloned alkaline protease gene of Geobacillus stearothermophilus.

Thermostable alkaline serine protease gene of Geobacillus stearothermophilus B-1172 was cloned and expressed in Escherichia coli BL21 (DE3) using pET-22b(+), as an expression vector. The growth conditions were optimized for maximal production of the protease using variable fermentation parameters, i.e., pH, temperature, and addition of an inducer. Protease, thus produced, was purified by ammonium sulfate precipitation followed by ion exchange chromatography with 13.7-fold purification, with specific activity of 97.5 U mg(-1) , and a recovery of 23.6%. Molecular weight of the purified protease, 39 kDa, was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was stable at 90 °C at pH 9. The enzyme activity was steady in the presence of EDTA indicating that the protease was not a metalloprotease. No significant change in the activity of protease after addition of various metal ions further strengthened this fact. However, an addition of 1% Triton X-100 or SDS surfactants constrained the enzyme specific activity to 34 and 19%, respectively. Among organic solvents, an addition of 1-butanol (20%) augmented the enzyme activity by 29% of the original activity. With casein as a substrate, the enzyme activity under optimized conditions was found to be 73.8 U mg(-1) . The effect of protease expression on the host cells growth was also studied and found to negatively affect E. coli cells to certain extent. Catalytic domains of serine proteases from eight important thermostable organisms were analyzed through WebLogo and found to be conserved in all serine protease sequences suggesting that protease of G. stearothermophilus could be beneficially used as a biocontrol agent and in many industries including detergent industry.

Insights into AlphaFold's breakthrough in neurodegenerative diseases.

Neurodegenerative diseases (ND) are disorders of the central nervous system (CNS) characterized by impairment in neurons' functions, and complete loss, leading to memory loss, and difficulty in learning, language, and movement processes. The most common among these NDs are Alzheimer's disease (AD) and Parkinson's disease (PD), although several other disorders also exist. These are frontotemporal dementia (FTD), amyotrophic lateral syndrome (ALS), Huntington's disease (HD), and others; the major pathological hallmark of NDs is the proteinopathies, either of amyloid-ß (Aß), tauopathies, or synucleinopathies. Aggregation of proteins that do not undergo normal configuration, either due to mutations or through some disturbance in cellular pathway contributes to the diseases. Artificial Intelligence (AI) and deep learning (DL) have proven to be successful in the diagnosis and treatment of various congenital diseases. DL approaches like AlphaFold (AF) are a major leap towards success in CNS disorders. This 3D protein geometry modeling algorithm developed by DeepMind has the potential to revolutionize biology. AF has the potential to predict 3D-protein confirmation at an accuracy level comparable to experimentally predicted one, with the additional advantage of precisely estimating protein interactions. This breakthrough will be beneficial to identify diseases' advancement and the disturbance of signaling pathways stimulating impaired functions of proteins. Though AlphaFold has solved a major problem in structural biology, it cannot predict membrane proteins-a beneficial approach for drug designing.

Melatonin as a master regulatory hormone for genetic responses to biotic and abiotic stresses in model plant Arabidopsis thaliana: a comprehensive review.

Melatonin is a naturally occurring biologically active amine produced by plants, animals and microbes. This review explores the biosynthesis of melatonin in plants, with a particular focus on its diverse roles in Arabidopsis thaliana , a model species. Melatonin affects abiotic and biotic stress resistance in A. thaliana . Exogenous and endogenous melatonin is addressed in association with various conditions, including cold stress, high light stress, intense heat and infection with Botrytis cinerea or Pseudomonas , as well as in seed germination and lateral root formation. Furthermore, melatonin confers stress resistance in Arabidopsis by initiating the antioxidant system, remedying photosynthesis suppression, regulating transcription factors involved with stress resistance (CBF, DREB, ZAT, CAMTA, WRKY33, MYC2, TGA) and other stress-related hormones (abscisic acid, auxin, ethylene, jasmonic acid and salicylic acid). This article additionally addresses other precursors, metabolic components, expression of genes (COR , CBF , SNAT , ASMT , PIN , PR1 , PDF1.2 and HSFA ) and proteins (JAZ, NPR1) associated with melatonin and reducing both biological and environmental stressors. Furthermore, the future perspective of melatonin rich agri-crops is explored to enhance plant tolerance to abiotic and biotic stresses, maximise crop productivity and enhance nutritional worth, which may help improve food security.

Diagnostic Accuracy of Cell Block and Immunohistochemistry in Effusion Cytology.

Introduction Although the cytology of effusion fluids is considered a routine laboratory test, it has recently emerged as an essential tool in determining the primary site of origin of carcinoma of unknown primary. The sensitivity for diagnosing malignancy has enhanced with the inclusion of cytospin, cell block (CB), and immunohistochemistry (IHC) to effusion fluid cytology due to the improvement in morphological preservation and good cellular yield. The purpose of this study was to assess the diagnostic yield, sensitivity, specificity, positive predictive value, and negative predictive value of IHC and CB in effusion cytology. Methodology An institution-based cross-sectional study was conducted over a period of six months on 150 cases of effusion fluids submitted for diagnostic purposes. After the preparation of cytospin, the residual amount of centrifuged deposit was mixed with CytoLyt solution, thrombin, and plasma, and CBs were prepared. IHC was applied to the CB. Calretinin was used for mesothelial cells, and BerEP4, TTF-1, ER, WT-1, and CD-X2 were used for the confirmation and origin of malignant cells. Results The mean age of the patients was 51.75 ± 16.63 years. The male-to-female ratio was 1:1.24. Out of 150 cases, 78 were pleural effusions, 68 were peritoneal effusions, and four were pericardial effusions. Out of 150 cases, based on cytological examination alone, 66 (44%) were classified as benign, 27 (18%) as malignant, and 57 (38%) were suspicious for malignancy. When cytology was combined with CB and IHC, the diagnostic yield was increased to benign 95 (63.33%), malignant 48 (32%), and suspicious for malignancy 7 (4.67%). The most common cause of malignant pleural effusion was breast carcinoma in females and lung carcinoma in males. The most common primary tumor in malignant peritoneal effusion was ovarian carcinoma in females and colonic adenocarcinoma in males. The sensitivity and specificity of combined cytology with cell block and IHC were 92.31% and 98.95%, respectively. This combination produced significantly better results (p-value = 0.001) for detecting malignancy and reduced suspicious cases from 38% to 4%. Conclusion CB, in combination with IHC, increases the diagnostic yield and aids in detecting malignancy at an unknown primary site in effusion fluids. Both of these techniques can thus enhance the sensitivity and specificity of the diagnosis of effusion cytology. Hence, CB and IHC have advanced utility over cytological smears in effusion fluid cytological diagnosis.

Clinicopathological Spectrum of Benign Skin Adnexal Tumors in the Pakistani Population: A Single-Center Study.

Background Skin adnexal tumors (SATs) are categorized per the site of origin, for example, hair follicles, sebaceous glands, and sweat glands. In our population, there is limited information related to the clinicopathological characteristics of these tumors. Management and prognosis depend largely upon the morphological type of the tumor. In this study, we assessed the disease spectrum and most prevalent subtypes of benign SATs. Methodology An analysis of 565 cases was conducted in this cross-sectional study between January 2018 and December 2022, using a non-probability consecutive sampling approach. Patient age, gender, site of involvement, and diagnosis were documented according to the fourth edition of the WHO Classification of Skin Tumors published in 2018. Data were entered and analyzed using SPSS Version 26.0 (IBM Corp., Armonk, NY). Results Our study had 565 patients, out of which 271 (47.9%) were males and 294 (52.1%) were females. The mean age was 40.97±19.3 years (range, 2-100 years). Anatomical site variations were as follows, head and neck (n=336, 59.4%), extremities (n=124, 22%), trunk (n=84, 14.9%), and genital areas (n=21, 3.7%). The most common histological subtypes of benign SATs were sweat gland origin (n=350, 62.0%), followed by hair follicle origin (n=161, 28.5 %), and sebaceous gland origin (n=54, 9.5%). Conclusion Sweat gland tumors were the most prevalent class of benign SATs in our study, in which hidradenoma and poroma were the most frequent subtypes. Hair follicle origin was the second most prevalent class of tumors with pilomatricoma being the most frequent. Sebaceous tumors were overall uncommon; nevus sebaceous of Jadasshon was the most common tumor in this class.

Computational insights into missense mutations in HTT gene causing Huntington's disease and its interactome networks.

BACKGROUND: Huntington's disease is a rare neurodegenerative illness of the central nervous system that is inherited in an autosomal dominant pattern. Mutant huntingtin protein is produced as a result of enlargement of CAG repeat in the N-terminal of the polyglutamine tract. AIM OF THE STUDY: Herein, we aim to investigate the mutations and their effects on the HTT gene and its genetic variants. Additionally, the protein-protein interaction of HTT with other proteins and receptor-ligand interaction with the three-dimensional structure of huntingtin protein were identified. METHODS: A comprehensive analysis of the HTT interactome and protein-ligand interaction has been carried out to provide a global picture of structure-function analysis of huntingtin protein. Mutations were analyzed and mutation verification tools were used to check the effect of mutation on protein function. RESULTS: The results showed, mutations in a single gene are not only responsible for causing a particular disease but may also cause other hereditary disorders as well. Moreover, the modification at the nucleotide level also cause the change in the specific amino acid which may disrupt the function of HTT and its interacting proteins contributing in disease pathogenesis. Furthermore, the interaction between MECP2 and BDNF lowers the rate of transcriptional activity. Molecular docking further confirmed the strong interaction between MECP2 and BDNF with highest affinity. Amino acid residues of the HTT protein, involved in the interaction with tetrabenazine were N912, Y890, G2385, and V2320. These findings proved, tetrabenazine as one of the potential therapeutic agent for treatment of Huntington's disease. CONCLUSION: These results give further insights into the genetics of Huntington's disease for a better understanding of disease models which will be beneficial for the future therapeutic studies.

Enzymatic hydrolysis of low temperature alkali pretreated wheat straw using immobilized ß-xylanase nanoparticles.

A low temperature alkali (LTA) pretreatment method was used to treat wheat straw. In order to obtain good results, different factors like temperature, incubation time, NaOH concentration and solid to liquid ratio for the pretreatment process were optimized. Wheat straw is a potential biomass for the production of monomeric sugars. The objective of the current study was to observe the saccharification (%) of wheat straw with immobilized magnetic nanoparticles (MNPs). For this purpose, immobilized MNPs of purified ß-xylanase enzyme was used for hydrolysis of pretreated wheat straw. Wheat straw was pretreated using the LTA method and analyzed by SEM analysis. After completion of the saccharification process, saccharification% was calculated by using a DNS method. Scanning electron micrographs revealed that the hemicellulose, cellulose and lignin were partially removed and changes in the cell wall structure of the wheat straw had caused it to become deformed, increasing the specific surface area, so more fibers of the wheat straw were exposed to the immobilized ß-xylanase enzyme after alkali pretreatment. The maximum saccharification potential of wheat straw was about 20.61% obtained after pretreatment with optimized conditions of 6% NaOH, 1/10 S/L, 30 °C and 72 hours. Our results indicate the reusability of the ß-xylanase enzyme immobilized magnetic nanoparticles and showed a 15% residual activity after the 11th cycle. HPLC analysis of the enzyme-hydrolyzed filtrate also revealed the presence of sugars like xylose, arabinose, xylobiose, xylotriose and xylotetrose. The time duration of the pretreatment has an important effect on thermal energy consumption for the low-temperature alkali method.

Benign Cutaneous Cysts: A Comprehensive Analysis of 1160 cases.

Background Cysts are common skin abnormalities that are mostly benign; however, sometimes malignant lesions may present clinically as cystic manifestations. Benign cutaneous cysts can be of different morphological types and their diagnosis relies on histological evaluations. The most common mode of treatment is surgical excision, which is curative. Methodology This is a retrospective cross-sectional study conducted at the Department of Histopathology, Chughtai Institute of Pathology, Lahore, Pakistan from 1st January 2020 to 31st December 2022. Non-probability consecutive sampling was done, and all the cases of benign cutaneous cysts were included. All cases were microscopically reviewed by two histopathologists, and variables like age, gender, site of the lesion, and histological diagnosis were noted. The data were analyzed using IBM SPSS Statistics for Windows, Version 29 (Released 2022; IBM Corp., Armonk, New York, United States). Results A total of 1160 recorded cases of benign cutaneous cysts were included. Overall gender distribution revealed males (n=489, 42.1%) and females (n=671, 57.8%). The age range was 3 to 91 years with a mean age of 37.56 ± 16.05 years. The three most common cysts were epidermal inclusion cysts (74.3%), trichilemmal cysts (15.1%), and dermoid cysts (6.3%). Other cysts were uncommon including hidrocystoma (1.9%), steatocystoma (0.3%), verrucous cysts (0.3%), comedones (0.6%), hybrid cysts (0.2%), milia (0.3%), and vellus hair cysts (0.2%). The most common site was back (23.5%) for epidermal inclusion cysts, scalp (74.4%) for trichilemmal cysts, and eye (33.8%) for dermoid cysts. Conclusion Benign cutaneous cysts have a broad morphological spectrum with a wide age range. Epidermal inclusion cysts, trichilemmal cysts, dermoid cysts, and hidrocystoma account for the four most common types. For each of the other cyst type, the prevalence was under 1%. Female gender predominated in epidermal inclusion cysts, trichilemmal cysts, and dermoid cysts while male gender was common in other cysts. Overall majority of the cysts presented in the head and neck area.

Morphological Spectrum of Gastritis in Endoscopic Biopsies and Its Association With Helicobacter pylori Infection.

Introduction Helicobacter pylori (H. pylori) infection is the most common cause of gastritis. The consequences of the persistent infection range from acute ulceration to the development of gastric neoplasia. The purpose of the study is to determine the histopathological pattern of gastritis in gastric biopsies and its association with H. pylori. Materials and methods This is a retrospective study of all the gastric biopsies received in the Department of Histopathology, Chughtai Institute of Pathology, Lahore, Pakistan from January 12, 2021 to April 10, 2021. Sections were cut from formalin-fixed, paraffin-embedded (FFPE) tissue blocks. Slides were stained with routine, special and immunohistochemical stains. The cases were then blindly reviewed by two pathologists with a special interest in Gastrointestinal Pathology. Microscopic features based on updated Sydney classification were recorded. The collected data were then analyzed by using SPSS version 20 (IBM Corp., Armonk, NY). Results A total of 429 gastric biopsies were examined. Out of which, 202 (47.1%) were male and 227 (52.9%) were female with a female-to-male ratio of 1.2:1. Their ages ranged from 12 to 100 years and a peak in the fourth decade of life with a mean age of 43 years (median age 49 years). Among gastritis, activity was seen in 194 (45%) and chronicity in 237 (55%) cases. H. pylori was seen in 174 (40.5%) cases and there was a strong statistically significant association between H. pylori infection and severity of gastritis (p<0.001). Other parameters of Sydney classification, such as atrophic changes, were seen in 144 (33%) cases, and intestinal metaplasia in 10 (2.3%) cases. Conclusion H. pylori organism was the most common cause of gastritis in our environment. The severity of gastritis is directly related to the H. pylori infection. If neutrophils are identified on surface epithelium, then H. pylori should be searched with increased attention on morphology and/or on immunohistochemical stain of H. pylori. Chronic inflammation and H. pylori density can also guide treatment which is necessary to avoid complications.

Enzymatic hydrolysis of lignocellulosic biomass using a novel, thermotolerant recombinant xylosidase enzyme from Clostridium clariflavum: a potential addition for biofuel industry.

The present study describes the cloning, expression, purification and characterization of the xylosidase gene (1650 bp) from a thermophilic bacterium Clostridium clariflavum into E. coli BL21 (DE3) using the expression vector pET-21a(+) for utilization in biofuel production. The recombinant xylosidase enzyme was purified to homogeneity by heat treatment and immobilized metal ion affinity chromatography. SDS-PAGE determined that the molecular weight of purified xylosidase was 60 kDa. This purified recombinant xylosidase showed its maximum activity at a temperature of 37 °C and pH 6.0. The purified recombinant xylosidase enzyme remains stable up to 90 °C for 4 h and retained 54.6% relative activity as compared to the control. The presence of metal ions such as Ca2+ and Mg2+ showed a positive impact on xylosidase enzyme activity whereas Cu2+ and Hg2+ inhibit its activity. Organic solvents did not considerably affect the stability of the purified xylosidase enzyme while DMSO and SDS cause the inhibition of enzyme activity. Pretreatment experiments were run in triplicate for 72 h at 30 °C using 10% NaOH. Saccharification experiment was performed by using 1% substrate (pretreated plant biomass) in citrate phosphate buffer of pH 6.5 loaded with 150 U mL-1 of purified recombinant xylosidase enzyme along with ampicillin (10 µg mL-1). Subsequent incubation was carried out at 50 °C and 100 rpm in a shaking incubator for 24 h. Saccharification potential of the recombinant xylosidase enzyme was calculated against both pretreated and untreated sugarcane bagasse and wheat straw as 9.63% and 8.91% respectively. All these characteristics of the recombinant thermotolerant xylosidase enzyme recommended it as a potential candidate for biofuel industry.

Clinicopathological Features of Neuroendocrine Tumors in Gastroenteropancreatic Tract: A Single Center Study.

BACKGROUND: Gastroenteropancreatic neuroendocrine tumors (GEP-NETs) are a heterogeneous group of tumors with varying biological, functional, and clinical characteristics that develop from the gastroenteropancreatic tract's diffuse neuroendocrine system. The objective of this study is to determine the clinicopathological features of GEP-NETs at our facility. METHODOLOGY: A cross-sectional analysis of 87 biopsies and resection specimens from January 2020 to January 2022 was performed. The histopathological reports as well as patient's demographic and clinic pathological data were obtained. Two pathologists with a special interest in gastroenteropancreatic pathology blindly reviewed all cases. The tumor grade and stage were determined using the WHO classification (2019) and the AJCC TNM system (8th edition). The data were analyzed with SPSS version 22 (IBM Corp., Armonk, NY, USA). RESULTS: Of the total 87 patients, 49 (56.3%) were male. The age range was 11 to 80 years, with a mean of 45.7±16.4 and the majority (56.3%) were under 50 years. The most frequent symptom was abdominal pain (55.2%). The most common site of GEP-NETs was the appendix (21.8%), followed by the ileum (18.4%), with the majority of tumors being non-functional (96.5%). Furthermore, neuroendocrine tumor (NET) grade 1 accounts for 62% of the total, followed by NET grade 2 (24.1%), neuroendocrine carcinoma (NET) grade 3 (10.3%), and mixed neuroendocrine-non-neuroendocrine neoplasms (MINENs) (3.5%). Synaptophysin was found to be positive in 83.9% cases while Chromogranin A was positive in 39.1%. A pathologic tumor (pT) stage was determined in 47 resection specimens in our study and the most common stage was pT3 (36.1%). Nodal metastasis was found in 25.5% of patients. CONCLUSIONS: According to our study, appendix and ileum were the most common GEP-NETs sites. The tumor site and grade were shown to significantly correlate among the clinicopathological features but there was no discernible correlation between the tumor grade and the gender, age, or pathological tumor (pT) stage.

Heterologous expression, molecular studies and biochemical characterization of a novel alkaline esterase gene from Bacillus thuringiensis for detergent industry.

Present study was aimed to clone and express the esterase encoding gene from Bacillus thuringiensis in E. coli BL21. Purification of recombinant esterase enzyme was achieved up to 48.6 purification folds by ion exchange chromatography with specific activity of 126.36 U mg-1. Molecular weight of esterase enzyme was 29 kDa as measured by SDS-PAGE. Purified esterase enzyme showed stability up to 90% at 90 °C and remained stable in a wide pH range (8-11). Molecular docking strengthens the experimental results by showing the higher binding energy with p-NP-butyrate. Enzyme activity was found to be reduced by EDTA but enhanced in the presence of other metal ions. Enzyme activity was reduced with 1% SDS, PMSF, and urea but organic solvents did not show considerable impact on it even at higher concentrations. Purified recombinant esterase was also found to be compatible with commercial laundry detergents and showed very good stability (up to 90%). All these properties proved the esterase enzyme from B. thuringensis a significant addition in detergent industry.

Effective utilization of magnetic nano-coupled cloned ß-xylanase in saccharification process.

The ß-xylanase gene (DCE06_04615) with 1041 bp cloned from Thermotoga naphthophila was expressed into E. coli BL21 DE3. The cloned ß-xylanase was covalently bound to iron oxide magnetic nanoparticles coated with silica utilizing carbodiimide. The size of the immobilized MNPs (50 nm) and their binding with ß-xylanase were characterized by Fourier-transform electron microscopy (FTIR) (a change in shift particularly from C-O to C-N) and transmission electron microscopy (TEM) (spherical in shape and 50 nm in diameter). The results showed that enzyme activity (4.5 ± 0.23 U per mL), thermo-stability (90 °C after 4 hours, residual activity of enzyme calculated as 29.89% ± 0.72), pH stability (91% ± 1.91 at pH 7), metal ion stability (57% ± 1.08 increase with Ca2+), reusability (13 times) and storage stability (96 days storage at 4 °C) of the immobilized ß-xylanase was effective and superior. The immobilized ß-xylanase exhibited maximal enzyme activity at pH 7 and 90 °C. Repeated enzyme assay and saccharification of pretreated rice straw showed that the MNP-enzyme complex exhibited 56% ± 0.76 and 11% ± 0.56 residual activity after 8 times and 13 times repeated usage. The MNP-enzyme complex showed 17.32% and 15.52% saccharification percentage after 1st and 8th time usage respectively. Immobilized ß-xylanase exhibited 96% residual activity on 96 days' storage at 4 °C that showed excellent stability.

Efficient biomass saccharification using a novel cellobiohydrolase from Clostridium clariflavum for utilization in biofuel industry.

The present study describes the cloning of the cellobiohydrolase gene from a thermophilic bacterium Clostridium clariflavum and its expression in Escherichia coli BL21(DE3) utilizing the expression vector pET-21a(+). The optimization of various parameters (pH, temperature, isopropyl ß-d-1-thiogalactopyranoside (IPTG) concentration, time of induction) was carried out to obtain the maximum enzyme activity (2.78 ± 0.145 U ml-1) of recombinant enzyme. The maximum expression of recombinant cellobiohydrolase was obtained at pH 6.0 and 70 °C respectively. Enzyme purification was performed by heat treatment and immobilized metal anionic chromatography. The specific activity of the purified enzyme was 57.4 U mg-1 with 35.17% recovery and 3.90 purification fold. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the molecular weight of cellobiohydrolase was 78 kDa. Among metal ions, Ca2+ showed a positive impact on the cellobiohydrolase enzyme with increased activity by 115%. Recombinant purified cellobiohydrolase enzyme remained stable and exhibited 77% and 63% residual activity in comparison to control in the presence of n-butanol and after incubation at 80 °C for 1 h, respectively. Our results indicate that our purified recombinant cellobiohydrolase can be used in the biofuel industry.

Correction: Efficient biomass saccharification using a novel cellobiohydrolase from Clostridium clariflavum for utilization in biofuel industry.

[This corrects the article DOI: 10.1039/D1RA00545F.].

Evaluation of toxicity potential of cobalt in wheat irrigated with wastewater: health risk implications for public.

The use of wastewater in irrigation weakens the beneficial properties of the soil and leads to a threat to food safety standards. The present research was designed to explore the cobalt toxicity associated with the ingestion of wastewater irrigated wheat. Wheat plants of five different varieties were collected from 7 different sites of Punjab, Pakistan, which were irrigated with three different sources of water. The sampling was done in two cropping years. The cobalt values in water, soil and wheat samples (root, shoot, grain) ranged from 0.46 to 1.24 mg/l, 0.15 to 1.20, 0.29 to 1.30, 0.08 to 0.76 and 0.12 to 0.57 mg/kg, respectively. All the water samples showed high cobalt concentration than the maximum permissible value. However, all the soil and wheat plant samples were found within the maximum allowable range. The high cobalt concentration in irrigating water showed that the continuous usage of such type of water may lead to cobalt toxicity in living organisms with the passage of time and may results in severe health risks.