Gastroprotective potential and mechanisms of action of Hedera nepalensis

Hedera nepalensis (H. nepalensis) , belonging to the family Araliaceae, is a medicinal plant traditionally used to treat stomach problems. The current study investigated the gastroprotective potential and the mechanism of action of H. nepalensis in diclofenac-and ethanol-induced ulcer models. Anti-oxidant and lipid peroxidation inhibitory prospects of H. nepalensis were checked out by free radical scavenging assay and UV spectrophotometer respectively. Effect of H. nepalensis on the pH, gastric total acidity of gastric juice and protective effects of H. nepalensis against ulcer models have been examined. Histopathological studies have been carried out. The aqueous methanol extract of H. nepalensis (100 µg/mL) showed anti-oxidant (83.55%) and lipid peroxidation inhibitory (70.88%) potential at 1000 µg/mL; the extract had no buffer potential. The extract (400 mg/kg) significantly (81.12% and 63.46%) showed gastroprotective effect in diclofenac and ethanol-induced rat ulcer models respectively. Histopathological studies confirmed the biochemical findings. FTIR analysis showed the presence of carboxylic acid, alkanes, conjugated alkanes, aldehydes and alkyl-aryl ethers. Gallic acid, M-coumaric acid and quercetin were found by HPLC analysis. H. nepalensis exhibited significant protection against diclofenac and ethanol induced gastric damage by anti-oxidant and lipid peroxidation suppression effects suggesting potential broad utility in treatment of diseases characterized with gastric damage

Reconnoitering Mycobacterium tuberculosis lipoproteins to design subunit vaccine by immunoinformatics approach

Background: Tuberculosis is an aerosol transmitted disease of human beings caused by Mycobacterium tuberculosis (Mtb). The only available vaccine for Mtb is Bacillus Calmette-Guérin (BCG). Currently no alternative or booster is available for BCG. The objective of this predictive approach was based on binding of MHC-I and MHC-II and B cell epitopes of Mtb for mouse host.Methods: Immunoinformatics approach was used to design subunit vaccine (SV) by joining 8 MHC-I bindings, 6 MHC-II bindings, and 8 B-Cell epitopes with AAV, GPGPG, and KK amino acid linkers, respectively. The efficacy of the SV was enhanced through Mtb protein Rv3763 (LpqH, PDB ID= 4ZJM) as an adjuvant at the N-terminal of SV. The in silico analyses evaluated the SV to predict allergenicity, antigenicity, and physico-chemical properties.Results: Predictions revealed that SV is non-allergic and highly antigenic. The physico-chemical analysis showed that the SV was stable and basic in nature. The three-dimensional structure of SV was stable with a high binding affinity against the mouse TLR2 receptor. In silico cloning suggested the effective transformation of SV into the eukaryotic expression vector.Conclusion: This study permits preclinical validation of the designed SV in mouse host to confirm its immunogenic potential and efficacy, which will help in controlling tuberculosis.Keywords: Immunoinformatics; Docking; Subunit vaccine; Lipoprotein; Tuberculosis

Infección con Sarcoptes scabiei (Acari: Sarcoptidae) en conejos (Oryctolagus cuniculus): estudio de caso.

A partir de una sospecha de sarna sarcóptica en tres de cinco conejos albinos europeos (Oryctolagus cuniculus), mantenidos con fines experimentales, se hizo un examen general que reveló áreas multifocales de alopecia alrededor de los ojos, las fosas nasales y los labios. Se procesaron cortes de piel y un análisis de PCR con el objeto de identificar los ácaros. La histopatología de los cortes de piel mostró epidermis erupcionada y estrato córneo con una infiltración de células inflamatorias. El raspado o frotis de piel reveló la presencia de ácaros tanto en adultos como en huevos. Al examen microscópico, los ácaros adultos fueron identificados como Sarcoptes (S.) scabiei (Acari: Sarcoptidae). El análisis de muestras de los tres casos por PCR reveló una banda de 311 pb confirmando la infestación de S. scabiei en los conejos. La infección en conejos con S. scabiei puede representar un problema de salud pública de transmisión indirecta debida a la manipulación de conejos infestados. Hasta la presente, este es el primer reporte de infección por S. scabiei en conejos de Pakistán

Prevalence of Black Quarter in Association with Patho-Morphological Alterations in Affected Tissues at Different Zones of Punjab-Pakistan

Background: Blackleg or Black Quarter (BQ) is a serious bacterial disease caused by Clostridium chauvoei. It causes edematous and gaseous changes in skeletal muscles of animals. The study was designed to find prevalence of BQ in 6 districts of Punjab, Pakistan from June 2018 to June 2019.Methods: Animals were randomly monitored, and selected in each union council regardless of age, sex and species, but lameness and hyperthermia were the selection parameters. A proforma was used to record the experimental readings of each animal. A total of 1500 swab samples from bovines (cattle and buffaloes) were processed through conventional PCR and culture sensitivity tests to examine the comparative diagnostic efficacy and suitability of the test.Results: Sensitivity and specificity of culture tests were 87.6% and 34.19% respectively, whereas PCR demonstrated 81% and 47.3% results for the said aspects, respectively. Alpha toxin gene (Ccta) was amplified at 52.2oC yielding an amplicon size of 1400 base pairs. Area wise and season wise prevalence of each animal was also determined. The prevalence of BQ in cattle was significantly higher (P 0.05) in Lahore (0%) and Nankana Sahib (8%) which belonged to the canal irrigated zones.Conclusion: There was higher prevalence of BQ in different areas of Punjab, Pakistan that show the higher risk of disease transmission. A systematic surveillance system is essential to regularly monitor the disease incidence and prevalence in these areas of Pakistan.Keywords: Black Quarter; Prevalence; Pathological; Ruminants;  Zone

Development and Exploitation of KASP Assays for Genes Underpinning Drought Tolerance Among Wheat Cultivars From Pakistan

High-throughput genotyping for functional markers offers an excellent opportunity to effectively practice marker-assisted selection (MAS) while breeding cultivars. We developed kompetitive allele-specific PCR (KASP) assays for genes conferring drought tolerance in common wheat (Triticum aestivum L.). In total, 11 KASP assays developed in this study and five already reported assays were used for their application in wheat breeding. We investigated alleles at 16 loci associated with drought tolerance among 153 Pakistani hexaploid wheat cultivars released during 1953–2016; 28 diploid wheat accessions (16 for AA and 12 for BB) and 19 tetraploid wheat (AABB) were used to study the evolutionary history of the studied genes. Superior allelic variations of the studied genes were significantly associated with higher grain yield. Favored haplotypes of TaSnRK2.3-1A, TaSnRK2.3-1B, TaSnRK2.9-5A, TaSAP-7B, and TaLTPs-1A predominated in Pakistani wheat germplasm indicating unconscious pyramiding and selection pressure on favorable haplotypes during selection breeding. TaSnRK2.8-5A, TaDreb-B1, 1-feh w3, TaPPH-7A, TaMOC-7A, and TaPARG-2A had moderate to low frequencies of favorable haplotype among Pakistani wheat germplasm pointing toward introgression of favorable haplotypes by deploying functional markers in marker-assisted breeding. The KASP assays were compared with gel-based markers for reliability and phenotypically validated among 62 Pakistani wheat cultivars. Association analyses showed that the favorable allelic variations were significantly associated with grain yield-contributing traits. The developed molecular marker toolkit of the genes can be instrumental for the wheat breeding in Pakistan

Study protocol of DIVERGE, the first genetic epidemiological study of major depressive disorder in Pakistan

INTRODUCTION: Globally, 80% of the burdenof major depressive disorder (MDD) pertains to low- and middle-income countries. Research into genetic and environmental risk factors has the potential to uncover disease mechanisms that may contribute to better diagnosis and treatment of mental illness, yet has so far been largely limited to participants with European ancestry from high-income countries. The DIVERGE study was established to help overcome this gap and investigate genetic and environmental risk factors for MDD in Pakistan. METHODS: DIVERGE aims to enrol 9000 cases and 4000 controls in hospitals across the country. Here, we provide the rationale for DIVERGE, describe the study protocol and characterise the sample using data from the first 500cases. Exploratory data analysis is performed to describe demographics, socioeconomic status, environmental risk factors, family history of mental illness and psychopathology. RESULTS AND DISCUSSION: Many participants had severe depression with 74% of patients who experienced multiple depressive episodes. It was a common practice to seek help for mental health struggles from faith healers and religious leaders. Socioeconomic variables reflected the local context with a large proportion of women not having access to any education and the majority of participants reporting no savings. CONCLUSION: DIVERGE is a carefully designed case-control study of MDD in Pakistan that captures diverse risk factors. As the largest genetic study in Pakistan, DIVERGE helps address the severe underrepresentation of people from South Asian countries in genetic as well as psychiatric research

Study protocol of DIVERGE, the first genetic epidemiological study of major depressive disorder in Pakistan.

INTRODUCTION: Globally, 80% of the burdenof major depressive disorder (MDD) pertains to low- and middle-income countries. Research into genetic and environmental risk factors has the potential to uncover disease mechanisms that may contribute to better diagnosis and treatment of mental illness, yet has so far been largely limited to participants with European ancestry from high-income countries. The DIVERGE study was established to help overcome this gap and investigate genetic and environmental risk factors for MDD in Pakistan. METHODS: DIVERGE aims to enrol 9000 cases and 4000 controls in hospitals across the country. Here, we provide the rationale for DIVERGE, describe the study protocol and characterise the sample using data from the first 500 cases. Exploratory data analysis is performed to describe demographics, socioeconomic status, environmental risk factors, family history of mental illness and psychopathology. RESULTS AND DISCUSSION: Many participants had severe depression with 74% of patients who experienced multiple depressive episodes. It was a common practice to seek help for mental health struggles from faith healers and religious leaders. Socioeconomic variables reflected the local context with a large proportion of women not having access to any education and the majority of participants reporting no savings. CONCLUSION: DIVERGE is a carefully designed case-control study of MDD in Pakistan that captures diverse risk factors. As the largest genetic study in Pakistan, DIVERGE helps address the severe underrepresentation of people from South Asian countries in genetic as well as psychiatric research

General Zia ul Haq Legacy: Provisional Constitutional Order (PCO) March 24, 1981. An Analytical Study

The Provisional Constitutional Order popularly term as PCO is an Emergency and Extra Constitutional Order, in which wholly or partially suspends the Constitution of Pakistan. The PCO fulfills the legal vacuum and act as temporary constitution in abeyance or suspension. Mostly, the Martial Law Orders have been enforced in country on behalf of CMLA. Constitution is an essential element of democracy. The constitution is a set of principles for guided the state affairs; the representatives elected through the democratic process and possess right way to make amendments in the constitution and determining according to the mandate of public. The amendment in the constitution is sometimes indispensible under the prevailing circumstances, necessity and time. Such amendments were made with the guidance and principles of constitution and the procedure for the change has been clearly defined in the constitution. If the procedure would not adopt then amendment would be un- legitimized and deviation of the law. The elections held under the military umbrella were never admirable on international or national levels, but in country, the situation was quite contrary because whenever a dictator sensed the need of amendments in the constitution, he accomplished it without any hesitation