Effect of smoking cessation interventions on abstinence and tuberculosis treatment outcomes among newly diagnosed patients: a randomized controlled trial.

The study evaluates the effectiveness of smoking cessation interventions [Behavioral Change Communication (BCC) and Behavioral Change Communication plus bupropion (BCC+)] compared to conventional Directly Observed Therapy Short Course (DOT) treatment in improving pulmonary tuberculosis treatment outcomes and abstinence among newly diagnosed pulmonary tuberculosis (PTB) patients, highlighting the scarcity of robust experimental studies. The current randomized controlled trial, conducted at Ojha Institute of Chest Diseases between October 2017 and June 2019, randomized 292 patients who were current smokers with newly diagnosed pulmonary tuberculosis into three arms: control (n = 97), BCC (n = 97), and BCC+ (n = 98) arms. The outcomes of the interventions were compared in terms of favorable treatment outcomes and abstinence achieved at the end of 6 months. Baseline characteristics were compared between groups. Cox regression quantified the effect size of interventions for both outcome variables and reported as (crude and adjusted) hazard ratios with 95% confidence intervals (CI). No statistically significant difference was observed in baseline characteristics in each arm. Both BCC+ and BCC showed a statistically significant effect in achieving favorable PTB outcomes at 6 months (aHR 2.37, 95% CI 1.52-3.70 and aHR 2.34, 95% CI 1.51-3.60), as well as for abstinence from smoking at 6 months (BCC+: aHR 4.03, 95% CI 2.18-7.44 and BCC: aHR 3.87, 95% CI 2.12-7.05) compared to the control arm. Both BCC and BCC+ aided by pharmacologic agents such as bupropion when incorporated with conventional DOTs were found to be significantly effective in attaining favorable tuberculosis treatment outcomes as well as in attaining smoking abstinence at the end of the 6-month treatment.This study shows that adding smoking cessation programs (with or without extra drugs like bupropion) to standard Directly Observed Treatment Short Course (DOTs) treatment for people who have recently been diagnosed with pulmonary tuberculosis has a great positive impact on how well the overall antituberculosis treatment works. Our trial shows very promising results for such a combined therapy (DOTs and smoking cessation) in a country where the burden of both tuberculosis and smoking is very high.

Evaluation of blood minerals and oxidative stress changing pattern in prepartum and postpartum Achai and Holstein Friesian dairy cows.

This research focused on evaluating blood mineral levels and malondialdehyde (MDA) as an indicator of oxidative stress before and after giving birth in Achai and Holstein Friesian (HF) dairy cows. Blood samples were obtained from a total of 50 cows representing both breeds on the third, second, and first week prior to calving, as well as on the day of calving (day 0). Subsequently, samples were collected on the 1-3 weeks postpartum to allow a comprehensive evaluation of blood parameters throughout the peripartum period. Results showed a significant (p < .01) decrease in serum zinc (Zn) levels on the day of parturition in both breeds. Additionally, HF cows exhibited higher (p < .01) Zn levels on week 2 before giving birth. Interestingly, blood selenium (Se) concentration increased (p < .01) in HF cows on weeks 2 and 3 after calving. In contrast, Achai cows showed a rise (p < .01) in blood Se on week 3 of parturition. Blood copper (Cu) levels were higher (p < .01) on weeks 2 and 3 after parturition in HF cows and on third week before parturition compared to Achai cows, where serum Cu remained high on week 1 of parturition. The findings indicated that blood magnesium (Mg) was higher (p < .01) on third week before parturition in Achai cows and on weeks 2 and 3 after parturition. Serum calcium (Ca) was higher (p < .01) in both HF and Achai cows on weeks 2 and 3 after parturition, and lower (p < .01) on third week before giving birth in both breeds. The mean blood MDA levels in Achai cows were lower (p < .01) on weeks 1 to 3 postpartum or 3 weeks before giving birth. In HF cows, serum MDA increased (p < .01) just before 2 weeks of parturition and remained elevated until 3 weeks of parturition. Thus, both breeds exhibited a similar pattern of mineral fluctuations; yet, Achai cows demonstrated greater resilience to oxidative stress compared to HF cows during the transition period.

Cellulose nanocrystals boosted hydrophobically associated self-healable conductive hydrogels for the application of strain sensors and electronic devices.

Currently, hydrogel-based flexible devices become hot areas for scientists in the field of electronic devices, artificial intelligence, human motion detection, and electronic skin. These devices show responses to external stimuli (mechanical signals) and convert them into electrical signals (resistance, current, and voltage). However, the applications of the hydrogel-based sensor are hampered due to low mechanical properties, high time response, low fatigue resistance, low self-healing nature, and low sensing range. Herein, a strain sensing conductive hydrogel constructed from the CNCs (cellulose nanocrystal) reinforced, in which acrylamide and butyl acrylate work as hydrophilic and hydrophobic monomers respectively. The incorporation of CNCs in the polymeric system has a direct effect on their mechanical properties. The hydrogel having a high amount of CNCs (C4), its fracture stress and fracture strain reached 371.2 kPa and 2108 % respectively as well as self-healing of C4 hydrogel Broke at 499 % strain and bore 197 kPa stress. The elastic behavior of the hydrogels was confirmed by the rheological parameter frequency sweep and strain amplitude. Besides this our designed hydrogel shows an excellent response to deformation with conductivity 420 mS m-1, shows response to small strain (10 %) and large (400 %) strain, and has excellent anti-fatigue resistance with continuous stretching for 700 s at 300 % strain, with 140 msec response time, and gauge factor 7.4 at 750 % strain. The C4 hydrogel can also work as electronic skin when it is applied to different joints like the finger, elbow, neck, etc. The prepared hydrogel can also work as an electronic pen when it is worn to a plastic pen cover.

Fungal community dynamics associated with the outbreaks of sugarcane root rot disease.

Sugarcane is a critical sugar and bioenergy crop in China. However, numerous factors, including root rot disease, hamper its yield. Root rot disease is a severe agricultural issue, reducing yield and threatening sustainable crop production. The current study aimed to explore the fungal community structure, identify and characterize the primary pathogen for sugarcane root rot in Guangzhou, China. Eighty-nine samples of sugarcane root, stalk, rhizosphere soil, and irrigation water were collected from five sites in Guangzhou, China. Subsequently, 276 fungal strains were isolated to identify the primary pathogens. The five most common genera identified were Penicillium, Fusarium, Gongronella, Trichoderma, and Cladosporium. Fusarium was more prevalent in the infected soil samples than in healthy ones. Pathogenic assays of the strains revealed that the strain GX4-46 caused 80% of the disease. The strain was confirmed as Fusarium commune through phylogenetic and genome sequence analysis. Rhizosphere soil samples from different regional crops were collected to better understand the fungal community structure and the primary pathogen. We observed a significant presence of Fusarium in irrigation water, indicating that the root rot disease could originate from the irrigation water and then spread as a soil-borne disease. This research is pioneering and one of the most comprehensive investigations on the occurrence and prevalence of sugarcane root rot disease. This study will serve as a reference for expanding the sugarcane industry and a foundation for further exploration and control of root rot.IMPORTANCESugarcane, a significant economic crop, faces challenges due to root rot pathogens that accumulate each year in plants and soil through ratoon planting. This disrupts soil microbial balance and greatly impedes sugarcane industry growth. Symptoms range from wilting and yellowing leaves to stunted growth and reduced seedling tillers. The rhizosphere microbiota plays an important role in plant development and soil health. Little is known about root rot fungal community structure, especially in sugarcane. Here, we focused on exploring the main causative pathogen of root rot in the area alongside a detailed survey of the rhizosphere soil of different severity sugarcane cultivars and rotation crops of the region. To validate the findings, we also investigated the irrigation water of the area. Our study revealed Fusarium commune as the causative pathogen of root rot in the area, primarily originating from water and later as soil-borne. Using Trichoderma can control the disease effectively.

Tuning the Magnetic Behavior of Zinc Ferrite via Cobalt Substitution: A Structural Analysis.

Cobalt-doped zinc ferrite is a contemporary material with significant structural and magnetic characteristics. Our study explores the magnetic properties of cobalt-substituted zinc ferrite (ZnxCo1-xFe2O4), synthesized via a simple sol-gel method. By varying the cobalt ratio from 0 to 0.5, we found that zinc substitution impacts both the magnetization and lattice parameters. FTIR analysis suggested the presence of functional groups, particularly depicting an M-O stretching band, within octahedral and tetrahedral clusters. X-ray diffraction analysis confirmed the phase purity and cubic structure. The synthesized materials exhibited an average particle size of 24-75 nm. Scanning electron microscopy revealed the morphological properties, confirming the formation of truncated octahedral particles. In order to determine the stability, mass loss (%), and thermal behavior, a thermal analysis (thermogravimetric analysis (TGA)/differential thermal analysis (DTA)) was performed. The magnetic properties of the synthesized ferrites were confirmed via a vibrating sample magnetometer (VSM). Finally, the highest saturated magnetization and lowest coercivity values were observed with higher concentrations of the cobalt dopant substituting zinc. The synthesized nanomaterials have good stability as compared to other such materials and can be used for magnetization in the near future.

Effect of body weight at photostimulation on productive performance and welfare aspects of commercial layers.

OBJECTIVE: Due to current selection practices for increased egg production and peak persistency, the production profile, age at maturity, and body weight criteria for commercial layers are constantly changing. Body weight and age at the time of photostimulation will thus always be the factors that need to be adequately addressed among various production systems. The current study was carried out to determine the effects of pullets' body weight (low, medium, and heavy) on their performance, welfare, physiological response, and hormonal profile. METHODS: With regard to live weight, 150 16-week-old pullets were divided into three groups using a completely randomized design (CRD) and held until the 50th week. One-way analysis of variance was used to evaluate the data under the CRD, and the least significant difference test was used to distinguish between treatment means. RESULTS: In comparison to the medium and light birds, the heavy birds had higher body weight at maturity, an earlier age at maturity, and higher egg weight, eggshell weight, eggshell thickness, egg yolk index, breaking strength, egg surface area, egg shape index, egg volume, and hormonal profile except corticosterone. However, the medium and light birds had lower feed consumption rates per dozen eggs and per kilogram of egg mass than the heavy birds. Light birds showed greater body weight gain, egg production, and egg specific gravity than the other categories. At 20 weeks of age, physiological response, welfare aspects, and catalase were non-significant; however, at 50 weeks of age, all these factors-aside from catalase-were extremely significant. CONCLUSION: The findings of this study indicate that layers can function at lower body weights during photostimulation; hence, dietary regimens that result in lighter pullets may be preferable. Additionally, the welfare of the birds was not compromised by the lighter weight group.

Phenotype versus genotype discordant rifampicin susceptibility testing in tuberculosis: implications for a diagnostic accuracy.

IMPORTANCE: An accurate diagnosis of drug resistance in clinical isolates is an important step for better treatment outcomes. The current study observed a higher discordance rate of rifampicin resistance on Mycobacteria Growth Indicator Tube (MGIT) drug susceptibility testing (DST) than Lowenstein-Jenson (LJ) DST when compared with the rpoB sequencing. We detected a few novel mutations and their combination in rifampicin resistance isolates that were missed by MGIT DST and may be useful for the better management of tuberculosis (TB) treatment outcomes. Few novel deletions in clinical isolates necessitate the importance of rpoB sequencing in large data sets in geographic-specific locations, especially high-burden countries. We explored the discordance rate on MGIT and LJ, which is important for the clinical management of rifampicin resistance to avoid the mistreatment of drug-resistant TB. Furthermore, MGIT-sensitive isolates may be subjected to molecular methods of diagnosis for further confirmation and treatment options.

Demographic and clinical variables in the dengue epidemic in Punjab, Pakistan.

Objectives: To identify the latest trends in the clinical picture and severity of the disease, which will help better understand and manage dengue. Methodology: It was a cross-sectional, hospital-based study performed in the tertiary care hospitals of Punjab from August 21 to December 2022, in which serologically and polymerase chain reaction (PCR) confirmed patients with dengue infection, were enrolled. Demographic and clinical variables were recorded on a pre-tested Performa, processed and presented in frequency and percentages, and graphs were generated. Mean and standard deviation was used to present continuous variables. Results: Out of a total of 580 patients, 472 were diagnosed with Dengue Fever (DF) and 108 with Dengue Hemorrhagic Fever (DHF). About 79.31% of the patients were male and 20.69% were females. The mean age of patients was 32.5±9 years. Among the clinical features the percentage of high-grade fever, body aches, and vomiting were the highest. The liver function profile showed that serum bilirubin, Serum aspartate transaminase (AST), serum alanine transaminase (ALT,) and alkaline phosphatase (ALP) levels were markedly raised. Conclusion: This study showed that with time the trends in the presentation of dengue are slowly shifting, which will help us better manage the disease burden in the future.

Inclusion of the fungus Pleurotus florida in the diet affects performance and feed efficiency traits in calves: a case study on Ravi buffalo.

The objective of the study was to investigate the effects of inclusion of Pleurotus florida treated wheat straw in the total mixed rations (TMRs) on feed intake, growth performance, nutrient digestibility, and nitrogen retention in male buffalo calves. As a pilot study, four TMRs, i.e., TMR1 having 0% P. florida treated wheat straw (FTWS), TMR2 (20% FTWS), TMR3 (40% FTWS), and TMR4 (60% FTWS) with berseem hay as basal diet, were formulated. Sixteen Nili-Ravi male buffalo calves (aged 10-12 months, weighing 73 ± 2.50 kg) were divided into four equal groups and randomly assigned one of four TMRs. A significant increase (P < 0.05) was observed in all nutrients intake, their digestibility, weight gain, and nitrogen retention with TMRs incorporated with FTWS. Highest feed conversion ratio (FCR) of 2.63 was noted with TMR1-0% and the lowest FCR (1.80) with TMR4-60%, on the other hand. In conclusion, the TMR4 (60% FTWS) has the potential to increase the weight gain, nutrient digestibility, nitrogen retention, and feed efficiency in buffalo calves. Therefore, inclusion of 60% Pleurotus florida treated wheat straw is recommended as TMRs with berseem hay based basal diet for feeding buffaloes calves.

Alzheimer's Disease; Mechanism, Mutations, and Applications of Nano-Medicine.

BACKGROUND: In the past 10 years, significant progress has been made in understanding the pathogenic chain of events that causes Alzheimer's disease (AD). According to the most widely accepted concept, the production and aggregation of ß-amyloid (Aß) peptides play a critical role in AD. As a result, therapeutic intervention with these processes is the focus of intense research. The Aß peptide is cleaved by the α-secretase, ß-secretase, and γ-secretase enzymes in a region near the pathogenic amyloid precursor protein (APP) and mutations occurring site. METHODS: In the current review, a complete picture of the risk factors behind AD has been investigated. Mutations involved in AD progression have also been screened in various studies. RESULTS: Most of the mutations in the amyloid precursor protein (APP) can lead to the accumulation of APP oligomers in the brain, leading to AD. Several point mutations in APP can cause familial AD (FAD), including the Swedish mutation (K>M670/671N>L) and the A673>V mutation. The pathogenic A673>V mutation and Swedish mutation (M670>K/N671>L) are present in the same region of amyloid precursor protein (APP). However, the A673>T mutation has been shown to confer protection against AD. CONCLUSION: More investigations are needed from geographically distinct regions on mutations associated with AD development and applications of nanomedicines for better management of the disease burden in the future. Nanotechnology-produced metal nanoparticles (NPs) have gotten much attention because of their wide range of uses in the medicinal and agricultural industries. Nanomedicine containing potential phytochemicals, including GX-50 and curcumin conjugated with NPs, maybe a potential candidate for treating AD.

Novel mutations in structural proteins of dengue virus genomes.

BACKGROUND: Genomic characterization of the dengue virus (DENV) is useful for understanding its molecular evolution, transmission, pathogenicity and infectivity. The DENV genomic RNA encodes three structural proteins, capsid (C) envelope (E) and membrane (M) proteins mediating viral entry and assembly during host infection. The current study aims to explore the DENV serotypes and mutations in the E and M proteins. METHODS: Twenty-three samples of DENV-positive patients were processed and selected for whole genome sequencing (WGS) from the Punjab Province of Pakistan. RESULTS: Among the 23 WGS, 19 samples showed numerous mutations (BioProject ID PRJNA943555). DENV1 and DENV2 are the most prevalent serotypes. A total of 179 mutations were detected in the E protein, in which K203E, T88A, I114L, and I293T are novel. The I270L, T272A, S273L, and T277A were found in the "kl" ß-hairpin (aa 270-279). The M protein harbors 74 mutations, of which 24 were novel. Three prominent complementary regions in the prM and E protein complex formations include R6, E46, D47, D63, and D65 on 'pr' peptide, and E84, K64, and H244, K247 on E, remain conserved except R6C. To our knowledge, it is the first comprehensive study of mutations in structural proteins. CONCLUSION: Genomic epidemiology is critical for analyzing emerging mutations and designing new policies therapeutic efforts for future outbreaks.

Blood metabolic pattern and somatic cells dynamic in Achai and Holstein Friesian cows during transition period.

This study focused on assessing blood metabolites and somatic cell count (SCC) during the transition period in Achai and Holstein Friesian cows. Blood samples were collected from cows of both breeds on days -21, -14, -7 (pre-partum period), on day 0 (parturition day) and then on days 7, 14 and 21 (post-partum period). Additionally, milk samples were obtained on the day of parturition and on days 7, 14 and 21 post-partum for SCC assessment. Among Holstein cows, blood glucose levels showed a significant increase on day -21 before calving, and a decrease on the 14th and 21st days post-partum. Similarly, the blood triglycerides concentration exhibited a significant rise in Holstein cows 21 days before calving, while Achai cows experienced a significant decline in blood triglycerides on the day of parturition and throughout the entire post-partum period. Comparing Holstein and Achai cows, blood cholesterol was significantly higher in Holstein cows on day -21 before calving, whereas Achai cows had significantly lower blood cholesterol levels on the day of parturition. Regarding blood protein, Holstein cows exhibited significantly higher levels on day 14 post-partum, while Achai cows had lower protein levels 7 days before calving. The results further demonstrated that SCC was significantly elevated in Holstein cows on day 21 of parturition and lower on the day of parturition in Achai cows. Overall, the metabolic profile trend and SCC were comparable, yet the studied parameters were more pronounced in Holstein Friesian compared to Achai cows.

Urea molasses mineral block under various feeding systems improved nutrient digestibility, productive performance and blood biochemical profile of Yaks.

BACKGROUND: The study aimed to investigate the effect of urea molasses mineral blocks (UMMB) on nutrient digestibility, productive performance and blood biochemical profile of indigenous yaks under various feeding systems. A total of sixteen yaks were randomly divided into four groups (n = 4 animal per group) and offered the, following feeding systems: (A) stall feeding, (B), urea molasses mineral block (UMMB) + stall feeding, (C) yard feeding and (D) UMMB + yard feeding. Trial lasted for 40 days. RESULTS: Results showed that nutrients intake (g) and nutrient digestibility (%) of dry matter (DM), organic matter (OM), crude protein (CP), ether extract (EE) and crude fiber (CF) were significantly higher (p < 0.05) in stall and yard feeding groups with UMMB licking. Blood zinc, cobalt, hemoglobin (Hb), red blood cell (RBC), glucose and serum glutamate private transaminase (SGPT) significantly (p < 0.05) increased in stall and yard feeding with UMMB licking. Milk yield, Ca and monounsaturated fatty acid except milk composition improved significantly (p < 0.05) in stall and yard feeding groups with UMMB licking. CONCLUSION: It was concluded that feeding of UMMB improved utilization of low-quality roughages and best results were obtained from stall and yard feedings with UMMB licking as compared to other groups.

In Silico Investigation of a Chimeric IL24-LK6 Fusion Protein as a Potent Candidate Against Breast Cancer.

Targeted delivery of therapeutic anticancer chimeric molecules enhances the efficacy of drug by improving cellular uptake and circulation time. Engineering the molecules to facilitate the specific interaction between chimeric protein and its receptor is critical to elucidate biological mechanism as well as accuracy in modeling of complexes. A theoretically designed novel protein-protein interfaces can serve as a bottom-up method for comprehensive understanding of interacting protein residues. This study was aimed for in silico analyses of a chimeric fusion protein against breast cancer. The amino acid sequences of the interleukin 24 (IL-24) and LK-6 peptide were used to design the chimeric fusion protein via a rigid linker. The secondary and tertiary structures along with physicochemical properties by ProtParam and solubility were predicted using online software. The validation and quality of the fusion protein was confirmed by Rampage and ERRAT2. The newly designed fusion construct has a total length of 179 amino acids. The top-ranked structure from alpha fold2 showed 18.1 KD molecular weight by ProtParam, quality factor of 94.152 by ERRAT, and a valid structure by a Ramachandran plot with 88.5% residues in the favored region. Finally, the docking and simulation studies were performed using HADDOCK and Desmond module of Schrodinger. The quality, validity, interaction analysis, and stability of the fusion protein depict a functional molecule. The fusion gene IL24-LK6 after cloning and expression in a suitable prokaryotic cell might be a useful candidate for developing a novel anticancer therapy.

Malocclusion: Prevalence and Determinants among Adolescents of Karachi, Pakistan.

OBJECTIVES: The aim of this study was to determine the prevalence of malocclusion and its associated demographic and clinical factors in young adolescents (13-15 years) of Karachi, Pakistan. MATERIALS AND METHODS: An epidemiological survey included 500 young adolescents of registered schools, madrassas (Islamic education system), and shop workers of Gulshan-e-Iqbal Town. It was a cross-sectional analytical study design. Multistage random sampling technique was used to enroll participants. The pattern of occlusion was recorded with other related features using Angle's classification. Health status was recorded through World Health Organization-guided indices (decayed, missing, and filled permanent teeth [DMFT], community periodontal index of treatment needs [CPITN], and body mass index [BMI]). The information, thus, obtained was analyzed through SPSS using the chi-squared test and regression models. RESULTS: Forty four percent of the participants were female, while overall estimated prevalence of malocclusion in young adolescents of Karachi was 57.4%. After adjustments, participants going to any kind of education system had less malocclusion in comparison to those who were not going to any education system (adjusted odds ratio [aOR] = 0.305, 95% confidence interval [CI] = 0.12-0.73); mother's education especially higher level (aOR = 2.02, 95% CI = 1.08-3.75) and presence of periodontal disease (aOR = 1.57, 95% CI = 1.06-2.33) were significantly associated with malocclusion. CONCLUSION: This study showed that the class I malocclusion is prevalent in the local community. Demographic factors like gender, age, self-reported ethnicity, and BMI did not show any significant role. Education or knowledge of parents and young adolescents does play an influential role in decreasing malocclusion. Young adolescents, who are more prone to oral health problems at an early age, would have more chances to develop occlusal discrepancies.

Precision agricultural robotic sprayer with real-time Tobacco recognition and spraying system based on deep learning.

Precision agricultural techniques try to prevent either an excessive or inadequate application of agrochemicals during pesticide application. In recent years, it has become popular to combine traditional agricultural practices with artificial intelligence algorithms. This research presents a case study of variable-rate targeted spraying using deep learning for tobacco plant recognition and identification in a real tobacco field. An extensive comparison of the detection performance of six YOLO-based models for the tobacco crop has been performed based on experimentation in tobacco fields. An F1-score of 87.2% and a frame per second rate of 67 were achieved using the YOLOv5n model trained on actual field data. Additionally, a novel disturbance-based pressure and flow control method has been introduced to address the issue of unwanted pressure fluctuations that are typically associated with bang-bang control. The quality of spray achieved by attenuation of these disturbances has been evaluated both qualitatively and quantitatively using three different spraying case studies: broadcast, and selective spraying at 20 psi pressure; and variable-rate spraying at pressure varying from 15-120 psi. As compared to the broadcast spraying, the selective and variable rate spray methods have achieved up to 60% reduction of agrochemicals.

Novel Mutations in MPT64 Secretory Protein of Mycobacterium tuberculosis Complex.

Tuberculosis (TB) is a global health problem caused by the Mycobacterium tuberculosis complex (MTBC). These bacteria secrete various proteins involved in the pathogenesis and persistence of MTBC. Among the secretory proteins, MPT64 (Rv1980C) is highly conserved and is also known as a major culture filtrate that is used in rapid diagnosis of MTBC. In the current study, we aimed to find the mutation in this highly conserved protein in isolates from the Pashtun-dominant province of Pakistan. We analyzed 470 M. tuberculosis whole-genome sequences of Khyber Pakhtunkhwa Province. Mutations in the MPT64 gene were screened through TB-Profiler and BioEdit software tools. The DynaMut web server was used to analyze the impact of the mutation on protein dynamics and stability. Among 470 MTB genomes, three non-synonymous mutations were detected in nine isolates, and one synonymous mutation (G208A) was found in four isolates. Mutation G211T (F159L), which was detected at the C-terminal domain of the protein in six isolates, was the most prominent. The second novel mutation, T480C (I70V), was detected in two isolates at the C-terminal side of the protein structure. The third novel mutation, A491C (L66R), was detected in a single isolate at the N-terminal side of the MPT64 protein. The effect of these three mutations was destabilizing on the protein structure. The molecular flexibility of the first two mutations increased, and the last one decreased. MPT64 is a highly conserved secretory protein, harboring only a few mutations. This study provides useful information for better managing the diagnosis of MTB isolates in high TB-burden countries.

Diversity and novel mutations in membrane transporters of Mycobacterium tuberculosis.

Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis (TB), encodes a family of membrane proteins belonging to Resistance-Nodulation-Cell Division (RND) permeases also called multidrug resistance pumps. Mycobacterial membrane protein Large (MmpL) transporters represent a subclass of RND transporters known to participate in exporting of lipid components across the cell envelope. These proteins perform an essential role in MTB survival; however, there are no data regarding mutations in MmpL, polyketide synthase (PKS) and acyl-CoA dehydrogenase FadE proteins from Khyber Pakhtunkhwa, Pakistan. This study aimed to screen mutations in transmembrane transporter proteins including MmpL, PKS and Fad through whole-genome sequencing (WGS) in local isolates of Khyber Pakhtunkhwa province, Pakistan. Fourteen samples were collected from TB patients and drug susceptibility testing was performed. However, only three samples were completely sequenced. Moreover, 209 whole-genome sequences of the same geography were also retrieved from NCBI GenBank to analyze the diversity of mutations in MmpL, PKS and Fad proteins. Among the 212 WGS (Accession ID: PRJNA629298, PRJNA629388, and ERR2510337-ERR2510345, ERR2510546-ERR2510645), numerous mutations in Fad (n = 756), PKS (n = 479), and MmpL (n = 306) have been detected. Some novel mutations were also detected in MmpL, PKS and acyl-CoA dehydrogenase Fad. Novel mutations including Asn576Ser in MmpL8, Val943Gly in MmpL9 and Asn145Asp have been detected in MmpL3. The presence of a large number of mutations in the MTB membrane may have functional consequences on proteins. However, further experimental studies are needed to elucidate the variants' effect on MmpL, PKS and FadE functions.

Genetic mutations underlying isoniazid-resistant Mycobacterium tuberculosis in Khyber Pakhtunkhwa, Pakistan.

Tuberculosis, caused by Mycobacterium tuberculosis, is a major public health issue in Pakistan. Isoniazid is a first-line pro-drug that requires activation through an enzyme called catalase peroxidase, but is subject to widespread resistance, driven by mutations in katG and inhA genes and other loci with compensatory effects (e.g., ahpC). Here, we used whole genome sequencing data from 51 M. tuberculosis isolates collected from Khyber Pakhtunkhwa province (years 2016-2019; all isoniazid phenotypically resistant) to investigate the genetic diversity of mutations in isoniazid candidate genes. The most common mutations underlying resistance were katG S315T (37/51), fabG1 -15C>T (13/51; inhA promoter), and inhA -154G>A (7/51). Other less common mutations (n < 5) were also identified in katG (R128Q, V1A, W505*, A109T, D311G) and candidate compensatory genes ahpC (-54C>T, -51G>A) and oxyS (M249T). Using DynaMut2 software, the mutants exhibited various degrees of stability and flexibility on protein structures, with some katG mutations leading to a decrease in KatG protein flexibility. Overall, the characterisation of circulating isoniazid resistant-linked mutations will assist in drug resistant TB management and control activities in a highly endemic area of Pakistan.

Room-Temperature Ferromagnetism in Cu/Co Co-Doped ZnO Nanoparticles Prepared by the Co-Precipitation Method: For Spintronics Applications.

In current work, pure ZnO and Zn0.96-x Cu0.04Co x O (0 ≤ x ≤ 0.05) nanoparticles were synthesized by the co-precipitation method. Structural analysis and phase determination of the formed nanoparticles was carried out using X-ray diffraction (XRD) and Williamson-Hall plots. The hexagonal wurtzite structure was manifested by all the samples with divergent microstructures. The change in lattice parameters, bond length, dislocation density, and lattice strain indicates that Cu and Co were successfully incorporated. Average crystallite size was found to be in the range of 32.16-45.42 nm for various doping concentrations. Field emission scanning electron microscopy results exhibited that the surface morphology is an amalgam of spherical-like and hexagon-like structures. Spherical-shaped grains were homogeneous and evenly distributed all over the structure. Fourier transform infrared spectra indicated that the absorption bands were blue-shifted with increasing Co concentration. The UV-visible absorption spectra showed high absorption in the UV region and weak absorption in the visible region. An increase in the energy band gap for the maximum absorption peak was observed from 3.49 eV for ZnO to 3.88 eV for Zn0.91Cu0.04Co0.05O. The Burstein-Moss effect explained the noticed blue shift in absorption spectra and energy band gaps. The vibrating sample magnetometer study revealed the change in the diamagnetic behavior of pure ZnO to the ferromagnetic behavior of the prepared nanoparticles at room temperature for different doping concentrations. In the current study, we have developed the room-temperature ferromagnetism (RTFM) for Cu and Co co-doped ZnO nanoparticles. Since RTFM is the key objective for dilute magnetic semiconductors, therefore it can be served as the desirable expectant for spintronics applications with improved functionalities and device concepts.