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Laccases are multi-copper oxidases and found in ligninolytic bacteria catalyzing the oxidation of both phenolic and non-phenolic compounds, however its application in lignin degradation suffers due to low oxidation rate, which have intensified the search for new laccases. In the present study, spore coat A protein (CotA) encoding gene having laccase like activity from Bacillus altitudinis SL7 (CotA-SL7) was cloned and expressed in Escherichia coli. The purified CotA-SL7 was active at wide range of temperature and pH with optimum activity at 55 °C and pH 5.0. The kinetic parameters of CotA-SL7 was determined with K m , V max , and k cat values 0.4 mM, 2777 μmol/min/mg, and 5194 s -1 , respectively. Molecular docking revealed the presence of Pro, Phe, Asp, Asn, His, and Ile residues at the active site taking part in the oxidation of ABTS. The purified CotA-SL7 reduced lignin contents by 31 % and changes in lignin structure were analyzed through fourier transformed infrared spectroscopy (FTIR), scanning electron microsscopy (SEM) and gas chromatography mass-spectrometry (GC-MS). The appearance of low molecular size compounds clearly indicates the cleavage of lignin polymer and opening of the benzene ring by purified CotA-SL7. Thus, high catalytic efficiency of CotA-SL7 makes it a suitable bio-catalyst for remediation of lignin contaminated wastewater from pulp and paper industries with clear insights into lignin degradation at molecular level., Competing Interests: Declaration of Competing Interest The authors have no conflict of interest., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Ticks are blood-feeding ecto-parasites that have a cosmopolitan distribution in tropical and subtropical regions of the world. Ticks cause economic losses in the form of reduced blood, meat and dairy products, as well as pathogen transmission. Different acaricides such as organochlorines, organophosphates, formamidines (e.g. amitraz), synthetic pyrethroids, macrocyclic lactones, fipronil, and fluazuron are currently used sequentially or simultaneously to control tick infestations. Most acaricide treatments now face increasingly high chances of failure, due to the resistance selection in different tick populations against these drugs. Acaricide resistance in ticks can be developed in different ways, including amino acid substitutions that result in morphological changes in the acaricide target, metabolic detoxification, and reduced acaricide entry through the outer layer of the tick body. The current literature brings a plethora of information regarding the use of different acaricides for tick control, resistance selection, analysis of mutations in target sites, and resistance mitigation. Alternatives such as synergistic use of different acaricides, plant-derived phytochemicals, fungi as biological control agents, and anti-tick vaccines have been recommended to avoid and mitigate acaricide resistance. The purpose of this review was to summarize and discuss different acaricides applied for tick control, their mechanisms of action and resistance selection, genetic polymorphisms in their target molecules, as well as the approaches used for diagnosis and mitigation of acaricide resistance, specifically in Rhipicephalus microplus ticks., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Obaid, Islam, Alouffi, Khan, da Silva Vaz, Tanaka and Ali.)

Despite being one of the most potent anticancer agents, cisplatin (CDDP) clinical usage is limited owing to the acquired resistance and severe adverse effects including nephrotoxicity. The current work has offered a unique approach by designing a pH-sensitive calcium carbonate drug delivery system for CDDP and oleanolic acid (OA) co-delivery, with an enhanced tumor efficacy and reduced unwanted effects. Micro emulsion method was employed to generate calcium carbonate cores (CDDP encapsulated) followed by lipid coating along with the OA loading resulting in the generation of lipid-coated cisplatin/oleanolic acid calcium carbonate nanoparticles (CDDP/OA-LCC NPs). In vitro biological assays confirmed the synergistic apoptotic effect of CDDP and OA against HepG2 cells. It was further verified in vivo through the tumor-bearing nude mice model where NPs exhibited enhanced satisfactory antitumor efficacy in contrast to free drug solutions. In vivo pharmacokinetic study demonstrated that a remarkable long circulation time with a constant therapeutic concentration for both drugs could be achieved via this drug delivery system. In addition, the in vivo imaging study revealed that DiR-loaded NPs were concentrated more in tumors for a longer period of time as compared to other peritoneal tissues in tumor bearing mice, demonstrating the site specificity of the delivery system. On the other hand, hematoxylin and eosin (H&E) staining of Kunming mice kidney tissue sections revealed that OA greatly reduced CDDP induced nephrotoxicity in the formulation. Overall, these results confirmed that our pH-sensitive dual loaded drug delivery system offers a handy direction for effective and safer combination chemotherapy., Competing Interests: The author declares no competing financial interest., (This journal is © The Royal Society of Chemistry.)

Background: Signal transducers and activators of the transcription (STAT) family is composed of seven structurally similar and highly conserved members, including STAT1, STAT2, STAT3, STAT4, STAT5a, STAT5b, and STAT6. The STAT3 signaling cascade is activated by upstream kinase signals and undergoes phosphorylation, homo-dimerization, nuclear translocation, and DNA binding, resulting in the expression of target genes involved in tumor cell proliferation, metastasis, angiogenesis, and immune editing. STAT3 hyperactivation has been documented in a number of tumors, including head and neck, breast, lung, liver, kidney, prostate, pancreas cancer, multiple myeloma, and acute myeloid leukemia. Drug discovery is a timeconsuming and costly process; it may take ten to fifteen years to bring a single drug to the market. Machine learning algorithms are very fast and effective and commonly used in the field, such as drug discovery. These algorithms are ideal for the virtual screening of large compound libraries to classify molecules as active or inactive., Objective: The present work aims to perform machine learning-based virtual screening for the STAT3 drug target., Methods: Machine learning models, such as k-nearest neighbor, support vector machine, Gaussian naïve Bayes, and random forest for classifying the active and inactive inhibitors against a STAT3 drug target, were developed. Ten-fold cross-validation was used for model validation. Then the test dataset prepared from the zinc database was screened using the random forest model. A total of 20 compounds with 88% accuracy was predicted as active against STAT3. Furthermore, these twenty compounds were docked into the active site of STAT3. The two complexes with good docking scores as well as the reference compound were subjected to MD simulation. A total of 100ns MD simulation was performed., Results: Compared to all other models, the random forest model revealed better results. Compared to the standard reference compound, the top two hits revealed greater stability and compactness., Conclusion: In conclusion, our predicted hits have the ability to inhibit STAT3 overexpression to combat STAT3-associated diseases., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

In this technologically developed scenario, many organizations in developing countries including Pakistan have expanded the enthusiasm for understanding and creating an encouraging administrative and managerial environment. Numerous organizations are struggling for structural changes by deserting the old-fashioned organizational management style and implementing an empowering leadership where leaders give more authority to subordinates in decision making and responsibilities with the aim to increase organizational productivity. Therefore, the study examined the leadership empowering behaviour as a predictor of employees' psychological well-being of the educational institutions at secondary level in Kohat Division, Pakistan. A total sample of 564 secondary school teachers (male n = 379; female n = 185) was carefully chosen through a stratified random sampling technique. In this study, a non-experimental predictive correlational design was adopted. In order to collect data from the participants, two different standardized research tools i.e., the Leader Empowering Behaviour Questionnaire and Ryff's Psychological Well-being Scale were used. After the collection of data, it was analyzed on the basis of mean, standard deviation, Pearson's product-moment correlation, and multiple linear regression model. In conclusion, the study confirmed a significant positive correlation between leadership empowering behaviour and employees' psychological well-being. Leadership empowering behaviours predict employees' psychological well-being positively. Therefore, it was recommended that empowering behaviour might be adopted by the school leaders to improve the employees' psychological well-being for better organizational productivity., Competing Interests: The authors have declared that no competing interests exist.

Objective: To assess the risk factors associated with tonsillitis., Methods: The cross-sectional study was conducted at Mardan Medical Complex and District Headquarter Hospital, Mardan, Pakistan, from January to June 2018, and comprised tonsillitis patients. Data was collected using a questionnaire which included different risk factors like age 1-10 years, gender, residential area, dietary habit etc. Data was analysed using SPSS 20., Results: Of the 325 subjects, 200(61.54%), were clinically diagnosed with tonsillitis; 138(69%) being males. Age, unhygienic living condition, balanced diet, stressful environment and the use of sore/spicy foods were identified as significantly associated factors (p<0.05)., Conclusions: Age, unhygienic living condition, balanced diet, stressful environment and the use of sore/spicy food were found to have a strong association with tonsillitis.

Objective: Sheep farming is an important source of livelihood in the southern Khyber Pakhtun Khwa province of Pakistan where they are reared under extensive system (ES). This system has limitations of improper feeding, rearing, and management which negatively affect their productivity. The present study compares different rearing systems and their stresses on fertility, behavior, and growth rate potential of two sheep breeds of Pakistan., Materials and Methods: Eighty ewes were selected; forty each from Damani and Balkhi breeds reared in semi-intensive system (SIS) and ES at Paharpur district, Dera Ismail Khan. Blood samples were collected in triplicate on days 0, 14, and 28 of May, and the molecular and behavioral stress, growth, and fertility rates were recorded., Results: The highest growth and fertility rate were observed in both the breeds in SIS than in ES ( p < 0.01). The values of stress indicators (cortisol, heat shock protein (HSP-70)) and behavioral stress parameters were found lower in Damani breed as compared to Balkhi breed in both rearing systems ( p < 0.01). All the molecular stress and behavioral stress parameters were negatively associated with metabolic hormones (T3 and T4) ( p < 0.01). A positive correlation was observed among all the molecular stress and behavioral stress parameters ( p < 0.01). Similarly, T3 and T4 were positively interlinked to one another ( p < 0.01)., Conclusion: It was concluded that both Damani and Balkhi sheep showed lower stress levels and better fertility and growth parameter in SIS than ES system of rearing. Local Damani breed exhibited well tolerant ability and genetic adaptation to the local environment as compared to Balkhi breed., Competing Interests: The authors declare no conflict of interest., (Copyright: © Journal of Advanced Veterinary and Animal Research.)

Coxiellosis is a zoonotic disease caused by the obligate intracellular bacterium Coxiella burnetii affecting the productive and reproductive capabilities of animals. This study was conducted to gain insight into the seroprevalence of coxiellosis in small ruminants in seven farms of the Punjab, Pakistan. Potential risk factors were assessed. In total, 1000 serum samples (500 from sheep and 500 from goats) and 163 ticks were collected from the ruminants. All these 163 ticks were merged into 55 pools (29 pools for ticks from sheep and 26 pools for ticks from goat). Serum samples were investigated using an indirect ELISA and PCR. Coxiella burnetii DNA was detected in 29 pooled seropositive samples and 11 pooled ticks by real-time qPCR. Serological analysis revealed a prevalence of 15.6% and 15.0% in sheep and goats, respectively. A significant association was found between seropositivity and different variables like district, lactational status, reproductive status, body condition and reproductive disorders. Univariate analysis showed that detection of C. burnetii DNA in tick pools was significantly associated with the presence of ticks on sheep and goats. However, a non-significant association was found for the prevalence of C. burnetii DNA in serum pools. Hence, C. burnetii infection is prevalent in small ruminants and ticks maintained at livestock farms in Punjab, Pakistan.

Background: The supra-condylar humeral fractures are commonest fractures at the elbow in children and represent a third of all limb fractures in children under the age of 7 years. These fractures are associated with neurological deficit and any of the three long nerves of upper arm can be involved in these fractures. This study was conducted to determine the frequency of median nerve injury in children aged 2-11 years presenting with closed supracondylar fracture of humerus., Methods: This descriptive cross-sectional study was conducted at the department of orthopaedics and trauma, Ayub Teaching Hospital Abbottabad from July 2016 to June 2018. One hundred and seventy-one patients with supracondylar fracture of humerus were enrolled. They were treated as per department protocols and the outcome in terms of neurological damage was noted for each study participant., Results: The frequency of neurological damage was 21.05% with median nerve affected most commonly (13.45%). A statistically strong association was found between damage to median nerve and type of fracture, arm involvement and sex of patients (p<0.05) while no statistically significant association with age was obtained., Conclusion: Neurological damage is a common complication of supracondylar humeral fractures with median nerve being the most commonly affected nerve in these fractures.

Objective: High ambient temperature in poultry is a challenging and fatal stress among environmental factors. It affects the production quality, damages the liver, and increases mortality in broilers. The present study is focused to explore appropriate utilization of Selenium (Se) as a feed additive in broiler chickens against high temperature., Materials and Methods: Day-old male broiler chickens (Ross 308) ( n = 200) were grouped according to the supplements used in their basal diets such as: corn-soybean basal diet as control (Con), a basal diet containing sodium selenite, basal diet with probiotics, and a basal diet containing selenium-enriched probiotics (SP). At the end of the experimental period of 42 days, the liver was isolated and was used to determine the antioxidant capacity through a spectrophotometer. Inflammatory and anti-inflammatory cytokines production in the liver was measured through a real-time polymerase chain reaction., Results: Hepatic analyses revealed the decreased level of malondialdehyde, whereas glutathione, glutathione peroxidase (GSH-Px), and superoxide dismutase levels were increased in the SP group. Furthermore, supplementation of SP significantly up-regulated the mRNA expression of glutathione peroxidase 1 (GPx1), GPx4, IL6, and IL10 and down-regulated the expression of pro-inflammatory cytokines., Conclusion: It is thus concluded that SP as a potential nutritive supplement may facilitate hepatic protection by suppressing hepatic oxidation, inflammation, and necrosis during the high ambient temperature of summer., Competing Interests: The authors declare that they have no conflict of interest., (Copyright: © Journal of Advanced Veterinary and Animal Research.)

We investigated the effects of selenium-enriched probiotics (SP) on broiler meat quality under high ambient temperature and explore their underlying mechanisms. A total of 200 1-day-old male broiler chicks (Ross 308) were randomly allotted to four treatment groups, each with five replicates, in groups of ten birds. These birds were fed a corn-soybean basal diet (C), a basal diet plus probiotics supplementation (P), a basal diet plus Se supplementation in the form of sodium selenite (SS, 0.30 mg Se/kg), and a basal diet with the addition of selenium-enriched probiotics (SP, 0.30 mg Se/kg). The experiment lasted for 42 days. The birds were sacrificed by cervical dislocation, and the breast muscles were removed for further process. Our results showed that SP diet significantly increased (p < 0.05) the physical (pH, colors, water holding capacity, drip loss, shear force) and sensory characteristics of breast meat. All P, SS, and SP supplementation enhanced the antioxidant system by increasing (p < 0.05) the Se concentrations, glutathione (GSH) levels, activities of glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) whereas decreasing (p < 0.05) malondialdehyde (MDA) levels, with SP being higher than P and SS. Moreover, SP diet significantly upregulated (p < 0.05) the mRNA levels of glutathione peroxidase genes (GPx1, GPx4) while it downregulated heat stress biomarkers such as heat shock protein (HSP) 70 as compared to C, P, and SS diets. In conclusion, our findings suggest that SP may function as beneficial nutritive supplement that is capable of improving meat quality during the summer season.

A total of 80 female albino mice were randomly allotted into five groups (n = 16) as follows: (A) normal control, (B) high-fat diet (HFD),; (C) HFD + probiotics (P), (D) HFD + sodium selenite (SS), and (E) HFD + selenium-enriched probiotics (SP). The selenium content of diets in groups A, B, C, D, and E was 0.05, 0.05, 0.05, 0.3, and 0.3 μg/g, respectively. The amount of probiotics contained in groups C and E was similar (Lactobacillus acidophilus 0.25 × 10(11)/mL and Saccharomyces cerevisiae 0.25 × 10(9)/mL colony-forming units (CFU)). The high-fat diet was composed of 15 % lard, 1 % cholesterol, 0.3 % cholic acid, and 83.7 % basal diet. At the end of the 4-week experiment, blood and liver samples were collected for the measurements of lipid metabolism, antioxidative status, histopathological lesions, and related gene expressions. The result shows that HFD significantly increased the body weights and liver damages compared to control, while P, SS, or SP supplementation attenuated the body weights and liver damages in mice. P, SS, or SP supplementation also significantly reversed the changes of alanine aminotransferase (AST), aspartate aminotransferase (ALT), total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), total protein (TP), high-density lipoprotein (HDL), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalasa (CAT), and malondialdehyde (MDA) levels induced by HFD. Generally, adding P, SS, or SP up-regulated mRNA expression of carnitine palmitoyltransferase-I (CPT1), carnitine palmitoyltransferase II (CPT2), acetyl-CoA acetyltransferase II (ACAT2), acyl-coenzyme A oxidase (ACOX2), and peroxisome proliferator-activated receptor alpha (PPARα) and down-regulated mRNA expression of fatty acid synthase (FAS), lipoprotein lipase (LPL), peroxisome proliferator-activated receptor gamma (PPARγ), and sterol regulatory element-binding protein-1 (SREBP1) involved in lipid metabolism. Among the group, adding SP has a maximum effect in improving lipid metabolism, antioxidative status, histopathological lesions, and related gene expression in mice fed a HFD.

Ochratoxin A (OTA) contamination is a worldwide problem in pig industry. The objectives of the present study were to investigate the toxicity of natural OTA in weaned piglets and to further explore the underlying mechanisms. Totally, 36 crossbred ([Landrace × Yorkshire] × Duroc) piglets were randomly divided into 3 groups (three replicates per group, 4 piglets per replicate), and fed a basal diet (Con group) and basal diets added with 0.4 mg (OTA-L group) or 0.8 mg OTA/kg (OTA-H group), respectively for 42 days. The results showed that growth performance was significantly decreased (P<0.05) in OTA added groups compared with Con group. OTA concentration was relatively high in serum and OTA concentration in kidney was higher than in liver, respectively. AST, creatinine and urea in serum of OTA added groups were significantly increased (P<0.05), while glucose, total protein, albumin and globulin in serum of OTA added groups were significantly decreased (P<0.05) compared with Con group. Degenerative changes were observed in the epithelial cells of proximal tubules and in hepatocytes of OTA added groups. Antioxidant capacities in blood of OTA added groups and in kidney of OTA-H group were significantly decreased (P<0.05) compared with Con group. The mRNA expressions of bcl-2 were up-regulated, mRNA expressions of bax were down-regulated and the ratio of bcl-2 and bax was increased in kidney and liver of OTA added groups compared with Con group. In conclusion, OTA could reduce antioxidant capacity and suppress apoptosis in tissues and cause degenerative changes in the epithelial cells in proximal tubules and hepatic cells, which may have a negative effect on the growth performance of piglets., (Copyright © 2015. Published by Elsevier GmbH.)

Pakistani wheat varieties are grown over a wide agro-climatic range and as such are anticipated to exhibit yield and quality differences. It is therefore necessary to investigate the nutritional status of wheat varieties in terms of biochemical and physiochemical characteristics available for food and nutritional purposes in Pakistan. The result shows that wheat grains of different varieties contain a net protein level of 9.15%-10.27%, 2.15%-2.55% total fats, 1.72%-1.85% dietary fibers, 77.65x10(-6)-84.25x10(-6) of potassium and 7.70x10(-6)-35.90x10(-6) of sodium ions concentration, 0.24x10(-6)-0.84x10(-6) of phosphorus, 1.44%-2.10% ash, 31.108-43.602 g of thousand grain mass (TGM) and 8.38%-9.67% moisture contents. This study is significant in providing an opportunity to explore the available wheat varieties and to further improve their nutritional excellence and also essential for setting nutritional regulations for domestic and export purposes.

Background: Humeral diaphyseal fracture usually heals with closed methods but when non union develops then it needs surgical intervention in the form of plating and bone grafting, intramedulary nailing (open or closed simple or interlocking nails) and external fixators (circular or one plane fixator). In our unit we treated non union humeral diaphyseal fracture with plating and bone grafting and shortening of fracture ends up to 4 to 5 cm when needed., Methods: This study was conducted at Orthopaedic Department of Ayub Teaching Hospital Abbottabad from January 2002 till December 2003. We included 15 cases with atrophic non-union in 9 (60%) and hypertrophic non-union in 6 (40%) patients. All atrophic non-union were treated with plating, bone shortening by transverse osteotomy and bone grafting, while hypertrophic non-union were treated with decortications of non-union ends and fixation with compression plates, with bone grafting in old age. Follow up measures were based on clinical (range of joints motion) and radiological (healing) findings. Follow up was done for up to 6 months., Results: Out of 15 patients the age range was 20-80 years, 12 (80%) were male and 03 (20%) female. Right humerus involved in 5 (33.33%) while left humerus in 10 (66.66%) patients. In 9 (60%) patients with atrophic non union bone shortening by transverse cut osteotomy was done while in remaining patients with hypertrophic non-union plating was done in 2 (13.33%) cases and plating with bone grafting in 4 (26.66%) patients. Union was achieved in all patients after 16 to 20 weeks of surgery. In one patient (6.66%) of 75 years age with hypertrophic non-union implant was loosened after 03 months of surgery. At that time healing (Union) was evident on X-rays and humeral brace was applied for further 03 months. Two patients (13.33%) got neuropraxia of radial nerve which resolved with in 3 months time. 02 patients (13.33%) developed shoulder stiffness which resolved after exercise., Conclusion: In Non Union of Humerus shortening by transverse osteotomy & rigid fixation with plates give excellent results in selected cases.

Background: Open reduction and rigid internal fixation has become the generally accepted method of treatment for displaced fractures of the olecranon in order to allow early mobilisation and to prevent contracture of the elbow. Comminuted fractures of olecranon are unstable, therefore, bone graft and tension band wiring are supposed to give good stability. Here we give an account of our experience with this procedure., Methods: We treated ten patients with comminuted fractures of the olecranon by multiple tension-band wiring and a graft from the iliac crest between 1999 and 2002 at Ayub Teaching Hospital, Abbottabad. After initial immobilization strengthening and endurance exercises were started. The patients were followed up for stability, muscle strength, active range of flexion and extension at elbow and rotation of forearm., Results: The time to union of the fractures was 3 to 7 months. No patient reported difficulties with activities of daily living or symptoms of instability of the elbow. The median flexion was up to 135 degrees (125 degrees to 145 degrees) with a median flexion contracture of 15 degrees (range 10 degrees to 30 degrees). The median pronation was 70 degrees (60 degrees to 80 degrees) and median supination 79 degrees (70 degrees to 90 degrees). Only three patients had mild pain and loss of strength. Five patients had excellent and 5 good results with a median Broberg and Morrey index score of 94.5 points (84 to 100)., Conclusion: Our results are in accordance with those reported from other centres and the technique is thought to be a practical alternative to plate fixation of olecranon fractures with extreme comminution.