Differential Combination of Cytokine and Interferon- γ +874 T/A Polymorphisms Determines Disease Severity in Pulmonary Tuberculosis

BACKGROUND: Mycobacterium tuberculosis infects nearly 1/3 of the world population and this reservoir forms the largest pool from which new cases arise. Among the cytokines, IFN-γ is a key determinant in protection against tuberculosis. Single nucleotide polymorphisms (SNPs) in IFN-γ gene (+874 T/A) which determine TT high ((hi)), AA low ((lo)) and TA intermediate ((int)) responder phenotypes have shown variable associations with tuberculosis disease outcome in different ethnic populations. The objective of the current study was to analyze IFN-γ gene combinations with other IFN-γ regulating cytokine genes (IL-10, TNF -α, IL-6) to see the effect of gene- combinations on disease severity outcome in pulmonary tuberculosis. METHODS AND FINDINGS: Study groups comprised of pulmonary TB patients stratified according to lung tissue involvement into mild (Pmd = 74) or advance (Pad = 23) lung disease and compared with healthy controls (TBNA = 166). Genotype analysis was carried out using amplification refractory mutation system-PCR (ARMS-PCR). IFN-γ gene (+874 T/A) functional SNP combinations in TNFα (-308 G/A), IL-10 (-1082 A/G) and IL-6 (-174 G/C) were analyzed. Single gene analysis (Pearson χ²) showed a dominant association of IFN-γ TT (hi) genotype (p = 0.001) and T allele (p = 0.001) with mild disease. IFN-γ(lo) -IL-10(lo) genotype combination was associated with advanced disease (p = 0.002). IFN-γ(hi) -IL-6(hi) combination was associated with mild disease (p = 0.0005) while IFN-γ(lo) -IL-6(int) was associated with protection against both forms of pulmonary disease (p = 0.002). CONCLUSION: Our results show that a limited number of IFN-γ gene combinations with other cytokine functional SNPs determine the outcome of disease severity in tuberculosis

Differential combination of cytokine and interferon- gamma +874 T/A polymorphisms determines disease severity in pulmonary tuberculosis.

Background:Mycobacterium tuberculosis infects nearly 1/3 of the world population and this reservoir forms the largest pool from which new cases arise. Among the cytokines, IFN-gamma is a key determinant in protection against tuberculosis. Single nucleotide polymorphisms (SNPs) in IFN-gamma gene (+874 T/A) which determine TT high ((hi)), AA low ((lo)) and TA intermediate ((int)) responder phenotypes have shown variable associations with tuberculosis disease outcome in different ethnic populations. The objective of the current study was to analyze IFN-gamma gene combinations with other IFN-gamma regulating cytokine genes (IL-10, TNF -alpha, IL-6) to see the effect of gene- combinations on disease severity outcome in pulmonary tuberculosis. Methods andFindings:Study groups comprised of pulmonary TB Patients stratified according to lung tissue involvement into mild (Pmd = 74) or advance (Pad = 23) lung disease and compared with healthy controls (TBNA = 166). Genotype analysis was carried out using amplification refractory mutation system-PCR (ARMS-PCR). IFN-gamma gene (+874 T/A) functional SNP combinations in TNFalpha (-308 G/A), IL-10 (-1082 A/G) and IL-6 (-174 G/C) were analyzed. Single gene analysis (Pearson chi) showed a dominant association of IFN-gamma TT (hi) genotype (p = 0.001) and T allele (p = 0.001) with mild disease. IFN-gamma(lo) -IL-10(lo) genotype combination was associated with advanced disease (p = 0.002). IFN-gamma(hi) -IL-6(hi) combination was associated with mild disease (p = 0.0005) while IFN-gamma(lo) -IL-6(int) was associated with protection against both forms of pulmonary disease (p = 0.002).Conclusion:Our results show that a limited number of IFN-gamma gene combinations with other cytokine functional SNPs determine the outcome of disease severity in tuberculosis

Strengthening lady health visitors and midwives for Sindh, Pakistan for non-communicable diseases (NCDs) prevention and management with refresh essential skills

They were addressing the rising burden of non-communicable diseases (NCDs), particularly in Low-Middle Income Countries (LMICs) like Pakistan. A proactive initiative undertaken by a Primary Health Service (PHS) in collaboration with a Private School of Nursing and Midwifery in Karachi conducted a 14-week project. 28 LHVs and Midwives (MWs) working in the community of PHS in various parts of Sindh, Pakistan were equipped with some common NCD prevention and management through online sessions and in-person refresher essential skills. The project aimed to equip them with the knowledge of early identification and management of some common NCDs and freshen some skills to promptly recognize and manage non-communicable diseases (NCDs) in the communities. The program included online sessions, hands-on simulation, and clinical exposure in a tertiary care hospital. Through a questionnaire, the participants gain knowledge regarding some common NCDs online sessions. With that, they feel confident after refreshing their skills in a simulation environment and clinical exposure. A participant’s feedback was that this type of session should planned once or twice a year. Nonetheless, this is one of the initiatives that can enhance the capabilities of LHVs and midwives in aligning with global efforts to combat the growing burden of NCDs and may improve community health outcomes

Quality Of Life of Women, Pre- and Post-Operative Breast Cancer Surgery

Objective: To evaluate the Quality of Life (QoL) of women with breast cancer who came for treatment in surgery department before diagnosis and post-operative time period. Methodology: A prospective cohort study was conducted at General surgery Department, Islamabad Medical complex, NESCOM, from October 2021 to March 2022. Seventy four diagnosed breast cancer patients, above 18 years of age, who underwent surgical treatment (MRM and Axillary clearance), were selected. QoL was assessed with the help of EORTC QLQ C-30 and EORTC BR-23 questionnaire. Data was collected on opd follow up and through telephone. SPSS 20 was used to analyze the data and Wilcoxon test and Kruskal-Wallis test were performed. Results: The QoL assessed at pre and post operative stage showed positive results only in the future prospects and emotional function domain. Whereas, negative results were scored in rest of the domains, which are symptoms in the arm, body image, financial concerns, sexual pleasure, cognitive function and physical function. Conclusion: The need for a multidisciplinary approach for breast cancer patients is required regarding different dimensions that can improve their QoL

A COMPARISON OF NALBUPHINE AND PENTAZOCINE IN CONTROLLING POSTOPERATIVE PAIN IN DOGS

Surgical success in most cases is governed by the quality of post-operative pain management. In Pakistan, most veterinary surgeons face a dire predicament as they fail astutely in this regard. Owing to the controlled dispensing of potent narcotics and their potential misuse, an imperative need for effective post-operative analgesic management of pain exists in dogs. 32 dogs were randomly divided into two groups. Group A was injected Nalbuphine @ 0.5 mg/kg post-operatively while Group B was injected Pentazocine @ 3mg/kg. Subjective and objective analysis of pain was conducted by unbiased observers. Vital signs (Temperature, pulse, respiration) were analyzed along with supplementation of hepatic and renal function tests. Objective and subjective analysis of both groups yielded results in the favor of pentazocine. In group A, temperature, pulse and respiration averaged 101.86±0.58oF, 83.46±2.75 per minute and 19.26±2.14 per minute respectively. Group 2 demonstrated temperature, pulse and respiration averages of 102.31±0.40oF, 83.41±2.74 per minute and 19.54±2.14 per minute respectively. Values of hepatic and renal function were also observed to be significantly higher in Nalbuphine treated group. All the results indicate that pentazocine is not only a significantly better analgesic but also has lower hepatotoxic and renal toxic effects

Alternate efflux pump mechanism may contribute to drug resistance in extensively drug-resistant isolates of mycobacterium tuberculosis

INTRODUCTION: Extensively drug-resistant tuberculosis (XDR-TB) has emerged as one of the biggest threats to public health and TB control programs worldwide. XDR-TB is caused by Mycobacterium tuberculosis (MTB) strains resistant to rifampin and isoniazid, as well as to a fluoroquinolone and to at least one injectable aminoglycoside. Drug resistance in MTB has primarily been associated with single nucleotide polymorphisms (SNPs) in particular genes. However, it has also been shown that efflux pumps may play a role in resistance of MTB. Upregulation of drug efflux pumps can decrease the intracellular concentration of drugs and reduce their efficacy. METHODS: Whole genome sequencing was performed on 32 XDR-TB clinical isolates. Sequence data were used to investigate SNPs in efflux pump genes as compared with the H37Rv reference genome. RESULTS: Of the XDR MTB strains, eight (21.62%) were wild type for rpsL, rrs (500 region), and gidB genes, but had non-synonymous (ns) SNPs (aspartic acid to histidine) in the drrA efflux pump gene at position 3273138. Three of eight (37.5%) XDR MTB strains, wild type for rpsL, rrs (500 region), gidB, and gyrB genes were phenotypically streptomycin sensitive and five (62.5%) XDR MTB strains were streptomycin resistant, while all XDR MTB strains, wild type for rpsL, rrs, gidB, and gyrB genes were resistant to fluoroquinolone (ofloxacin) and ethambutol. In addition, three XDR MTB strains wild type for rpsL, rrs, gidB, and drrA genes showed nsSNPs (isoleucine to valine) in the major facilitator superfamily, Rv1634 efflux pump gene at position 1839306. CONCLUSION: Our data show an nsSNP in the drrA efflux pump gene that may result in upregulation of drug efflux mechanisms in MTB strains. It is therefore imperative to understand the mechanism of efflux and its role in drug resistance, which will enable the identification of new drug targets and development of new drug regimens to counteract the drug efflux mechanism of MTB

Single nucleotide polymorphisms in efflux pumps genes in extensively drug resistant Mycobacterium tuberculosis isolates from Pakistan

It is challenging to understand mechanisms of drug resistance in Mycobacterium tuberculosis (MTB) due to the large variability in resistance associated genes. Efflux pump genes contribute to drug resistance and thus add to this complexity. Efflux pump gene protein superfamilies have been characterized by genome analysis of drug resistant strains and through invitro transcriptional studies. However, there is limited information regarding efflux pump genes in extensively drug resistant (XDR) tuberculosis (TB) isolates. Whole genome sequencing (WGS) based analysis of 37 extensively drug resistant (XDR) and five drug sensitive (DS) MTB clinical isolates was performed. Single nucleotide polymorphisms (SNPs) in efflux pump genes Rv0194, Rv1217, Rv1218, drrA, drrB, Rv1258, Rv1634, Rv2688, Rv1273, Rv1819, Rv1458, Rv1877 and Rv1250 were determined in the clinical isolates as compared with the H37Rv reference strain. Allele frequencies of SNPs identified in XDR strains were compared with DS strains. Gene expression of Rv0194, Rv2688, Rv1634, drrA and drrB was determined in XDR -TB isolates (n=9), DS-TB strains (n=4) and H37Rv. We identified SNPs in XDR-TB isolates which were either unique or present at very low frequencies in DS strains; Rv0194 G170V; Rv1217 L151R; Rv1258 P369T and G391R; Rv1273 S118G and I175T; Rv1877 I534T; Rv1250 V318X/A and S333A, and Rv2688 P156T. The expression of Rv2688 and drrB was found to be raised in XDR-TB as compared with DS-TB strains. We identified unique SNPs in efflux pump genes which may be associated with increased drug resistance in the isolates. Increased levels of Rv2688 and drrB efflux pump gene expression observed in XDR strains even in the absence of antibiotics suggests that these clinical isolates may be more refractory to treatment. Further studies are required to directly associate these mutations with increased resistance in MTB

Characterization of genomic variations in SNPs of PE_PGRS genes reveals deletions and insertions in extensively drug resistant (XDR) M. tuberculosis strains from Pakistan.

BACKGROUND: Mycobacterium tuberculosis (MTB) PE_PGRS genes belong to the PE multigene family. Although the function of PE_PGRS genes is unknown, it is hypothesized that the PE_PGRS genes may be associated with antigenic variability in MTB. MATERIAL AND METHODS: Whole genome sequencing analysis was performed on (n=37) extensively drug-resistant (XDR) MTB strains from Pakistan, which included Lineage 1 (East African Indian, n=2); Other lineage 1 (n=3); Lineage 3 (Central Asian, n=24); Other lineage 3 (n=4); Lineage 4 (X3, n=1) and T group (n=3) MTB strains. RESULTS: There were 107 SNPs identified from the analysis of 42 PE_PGRS genes; of these, 13 were non-synonymous SNPs (nsSNPs). The nsSNPs identified in PE_PGRS genes - 6, 9 and 10 - were common in all EAI, CAS, Other lineages (1 and 3), T1 and X3. Deletions (DELs) in PE_PGRS genes - 3 and 19 - were observed in 17 (80.9%) CAS1 and 6 (85.7%) in Other lineages (1 and 3) XDR MTB strains, while DELs in the PE_PGRS49 were observed in all CAS1, CAS, CAS2 and Other lineages (1 and 3) XDR MTB strains. All CAS, EAI and Other lineages (1 and 3) strains showed insertions (INS) in PE_PGRS6 gene, while INS in the PE_PGRSgenes 19 and 33 were observed in 20 (95.2%) CAS1, all CAS, CAS2, EAI and Other lineages (1 and 3) XDR MTB strains. CONCLUSION: Genetic diversity in PE_PGRS genes contributes to antigenic variability and may result in increased immunogenicity of strains. This is the first study identifying variations in nsSNPs and INDELs in the PE_PGRS genes of XDR-TB strains from Pakistan. It highlights common genetic variations which may contribute to persistence

Role of foliar spray of plant growth regulators in improving photosynthetic pigments and metabolites in Plantago ovata (Psyllium) under salt stress – A field appraisal

Salinity is one of the major abiotic factors that limit the growth and productivity of plants. Foliar application of plant growth regulators (PGRs) may help plants ameliorate the negative impacts of salinity. Thus, a field experiment was conducted at the Botanical Garden University of Balochistan, Quetta, to explore the potential role of PGRs, i.e., moringa leaf extract (MLE; 10%), proline (PRO; 1 µM), salicylic acid (SA; 250 µM), and thiourea (TU; 10 mM) in ameliorating the impacts of salinity (120 mM) on Plantago ovata, an important medicinal plant. Salinity hampered plant photosynthetic pigments and metabolites but elevated oxidative parameters. However, foliar application of PGRs enhanced photosynthetic pigments, including Chl b (21.11%), carotenoids (57.87%) except Chl a, activated the defense mechanisms by restoring and enhancing the metabolites, i.e., soluble sugars (49.68%), soluble phenolics (33.34%), and proline (31.47%), significantly under salinity stress. Furthermore, foliar supplementation of PGRs under salt stress led to a decrease of about 43.02% and 43.27% in hydrogen peroxide and malondialdehyde content, respectively. Thus, PGRs can be recommended for improved photosynthetic efficiency and metabolite content that can help to get better yield under salt stress, with the best and most effective treatments being those of PRO and MLE to predominately ameliorate the harsh impacts of salinity

Hyperglycemia-associated Alzheimer’s-like symptoms and other behavioral effects attenuated by Plumeria obtusa L. Extract in alloxan-induced diabetic rats

Diabetes mellitus is a chronic metabolic complaint with numerous short- and long-term complications that harm a person’s physical and psychological health. Plumeria obtusa L. is a traditional medicine used in the treatment of diabetes to reduce complications related to behavior. Plumeria is a genus with antipsychotic activities. The objective of this study was to examine the effects of a methanolic extract of Plumeria obtusa L. in the attenuation of diabetes, on symptoms of Alzheimer disease, and on other associated behavioral aspects. A single dose of alloxan was administered to an experimental group of rats to induce development of diabetes (150 mg/kg, intraperitoneal) and the rats were then administered selected doses of methanolic extract of Plumeria obtusa L. (Po.Cr) or glibenclamide (0.6 mg/kg) for 45 consecutive days. Behavioral effects were evaluated using three validated assays of anxiety-related behavior: the open field test, the light and dark test, and the elevated plus maze. Anti-depressant effects of Plumeria obtusa L. were evaluated using the forced swim test (FST) and memory and learning were assessed using the Morris water maze (MWM) task. Po.Cr was also evaluated for phytochemicals using total phenolic content (TPC), total flavonoid content (TFC), and high-performance liquid chromatography assays, and antioxidant capability was assessed through assays of DPPH radical scavenging, total oxidation capacity, and total reducing capacity. In the alloxan-induced model of diabetes, the administration of Po.Cr and glibenclamide for 45 days produced a marked decrease (p < 0.001) in hyperglycemia compared to control animals. Po.Cr treatment also resulted in improvement in indicators, such as body weight and lipid profile (p < 0.05), as well as restoration of normal levels of alanine transaminase (ALT) (p < 0.001), a biomarker of liver function. Diabetic rats presented more Alzheimer-like symptoms, with greater impairment of memory and learning, and increased anxiety and depression compared to non-diabetic normal rats, whereas treated diabetic rats showed significant improvements in memory and behavioral outcomes. These results demonstrate that Po.Cr reversed alloxan-induced hyperglycemia and ameliorated Alzheimer-related behavioral changes, which supports additional study and assessment of conventional use of the plant to treat diabetes and associated behavioral complications