The Effect of Obesity on Severity of Asthma: An Observational Prospective Study from Pakistan.

Objective: The current research study aimed to access the relationship between obesity and asthma exacerbations and severity among adult patients at the outpatient section of a federal hospital (PIMS) in Islamabad, Pakistan. Methods: A cross-sectional research study was carried out on 207 asthma adult patients belonging to different areas and ethnic groups from the country. The study setting was the PIMS hospital, which attracts patients from all over the country due to its facilities and cost-effective treatments. The body mass index (BMI) of asthma patients was calculated using the heights and weights of the study subjects. However, the pulmonary functions were calculated using a computerized spirometer i-e Spirolab III S/N 303681 in line with Winspiro PRO 7.1.version software. It presents the patient's forced vital capacity that expires in the first second of expiration to full (FEV1) in comparison to forced vital capacity (FVC) ratio, that is, Tiffeneau-Pinelli index was also recorded to determine the asthma severity. Results: According to recent surveys, the overall prevalence of patients with overweight and obesity was 29.0% and 23.7%, respectively. A Chi-square test was used, and a statistically significant relationship was observed between BMI and asthma severity (P < 0.001). The adult obese female patients presented poor pulmonary functions. The average FEV1/FVC ratio presented significant variance among four different categories of BMI with P < 0.05. This difference was due to the normal BMI category as the Tiffeneau-Pinelli index, that is, FEV1/FVC in the normal BMI group was significantly lower as compared to that in underweight and obese patients. Conclusion: The study subjects presented raised asthma severity in accordance with the raised BMI. Obese patients presented comparatively raised asthma exacerbations. Moreover, a statistically significant association of gender difference was observed between obesity and asthma severity. It was concluded that adult asthmatic women with obesity presented raised asthma severity as compared to adult asthmatic males.

Knowledge, beliefs and hesitancy among medical and non-medical university students toward COVID-19 vaccination in Punjab, Pakistan.

This study assessed the knowledge, beliefs, and hesitancy toward COVID-19 vaccination among medical (MS) and nonmedical students (NMS) in Punjab, Pakistan. An online cross-sectional survey was conducted by enrolling 624 MS and 476 NMS in the largest province of Pakistan, i.e., Punjab. Chi-square tests were used to determine significant frequency distributions, and logistic regression to determine associations. The majority of students, males and females, were between 18-25 years of age and had good self-reported health. MS demonstrated adequate knowledge and positive beliefs about COVID-19. Fewer MS were indecisive about the vaccination; nonetheless, overall, 71.5% of MS and 78.2% of NMS were willing to receive the vaccine. A greater number of MS students were concerned about vaccine safety or side effects. A greater number of NMS demonstrated misconceptions about the COVID-19 vaccine - its effect on fertility and alterations in genomic DNA. The binary logistics regression exhibited a significant association between students' knowledge in the medical sciences (OR;1.53,p = 0.002) and area of residence (OR;1.60,p = 0.008). Compared to NMS, MS had a better understanding of the COVID-19 vaccine and were concerned about the acceptability of the vaccine, although NMS had misconceptions about the COVID-19 vaccine. Knowledge was significantly associated with the field of study.

Population demographic history and population structure for Pakistani Nili-Ravi breeding bulls based on SNP genotyping to identify genomic regions associated with male effects for milk yield and body weight.

The domestic Nili-Ravi water buffalo (Bubalus bubalis) is the best dairy animal contributing 68% to total milk production in Pakistan. In this study, we identified genome-wide single nucleotide polymorphisms (SNPs) to estimate various population genetic parameters such as diversity, pairwise population differentiation, linkage disequilibrium (LD) distribution and for genome-wide association study for milk yield and body weight traits in the Nili-Ravi dairy bulls that they may pass on to their daughters who are retained for milking purposes. The genotyping by sequencing approach revealed 13,039 reference genome-anchored SNPs with minor allele frequency of 0.05 among 167 buffalos. Population structure analysis revealed that the bulls were grouped into two clusters (K = 2), which indicates the presence of two different lineages in the Pakistani Nili-Ravi water buffalo population, and we showed the extent of admixture of these two lineages in our bull collection. LD analysis revealed 4169 significant SNP associations, with an average LD decay of 90 kb for these buffalo genome. Genome-wide association study involved a multi-locus mixed linear model for milk yield and body weight to identify genome-wide male effects. Our study further illustrates the utility of the genotyping by sequencing approach for identifying genomic regions to uncover additional demographic complexity and to improve the complex dairy traits of the Pakistani Nili-Ravi water buffalo population that would provide the lot of economic benefits to dairy industry.

Prevalence of Toxoplasma gondii oocysts through Copro-PCR in cats at Pet Center (UVAS), Lahore, Pakistan.

Toxoplasmosis is a major zoonotic disease of warm-blooded animals caused by Toxoplasma gondii. Cats are the only definitive host and they excrete environmentally resistant T. gondii oocysts in their faeces. Coproscopy was used to detect oocysts of enteric coccidians and then Copro-PCR was employed to test specifically for T. gondii in 470 cat samples. The prevalence of T. gondii oocysts was 2.3% (11/470) based on PCR. We observed 15 (3.2%) of 470 samples positive for coccidian oocysts by microscopy. The presence of Copro-DNA of T. gondii was found significantly higher (p<0.05) in males than females. We tested 11 samples of T. gondii oocysts in which 9 samples were from coccidian oocysts positive samples and 2 samples from negative faecal samples. Our results showed that PCR is the reliable method for the detection of faecal oocysts of T. gondii in cats as compared to microscopy. As per our knowledge, ours is first study for Copro-PCR prevalence of cats' T. gondii oocysts excretion in Pakistan.

Induction of specific humoral immune response in mice immunized with ROP18 nanospheres from Toxoplasma gondii.

Toxoplasmosis is one of the most common zoonotic protozoal diseases. Recent advances in biotechnology have produced recombinant protein, which are immunogenic, and progress in nano-pharmaceutics has generated encapsulated protein in nanospheres, which are suitable for vaccine delivery. DNA was extracted from Toxoplasma gondii oocysts and was confirmed through nested PCR and sequencing. The 1665 bp of ROP18 was cloned into the easy vector system: pGEM-T by the T-A cloning method. DH5α bacteria were transfected with pGEM-ROP18. ROP18 was subcloned from pGEM-ROP18 into pET28-ROP18. BL21 bacteria were transfected with pET28-ROP18. Thus, rROP18 protein was expressed in BL21 bacteria by induction at different concentrations of isopropyl ß-D-1-thiogalactopyranoside. Protein expression was confirmed through SDS-PAGE and Western blotting. The immunoblot of rROP18 was recognized by anti-HIS antibodies and sera from infected mice at 67 kDa. Recombinant ROP18 protein was encapsulated in nanoparticles with PLGA and was characterized through scanning electron microscopy. Intraperitoneal immunizations with rROP18 protein and intranasal immunization of nanospheres were carried out in mice, and the immune response was detected by ELISA. Results showed that rROP18 in nanospheres administered intra-nasally elicited elevated responses of specific IgA and IgG2a as compared to groups inoculated intra-nasally with rROP18 alone, or injected subcutaneously with rROP18 in montanide adjuvant. It was concluded that nanospheres of ROP18 would be a non-invasive approach to develop vaccination against T. gondii. Further experiments are needed to determine the cellular response to these nanospheres in a mouse model for chronic toxoplasmosis.

DNA Amplification Techniques for the Detection of Toxoplasma gondii Tissue Cysts in Meat Producing Animals: A Narrative Review Article.

BACKGROUND: Toxoplasma gondii is an intracellular parasite, which infects one-third population of world. Humans and animals acquire infection by ingesting oocytes from feces of cats or by meat of other animals having cysts that may lead to congenital, ocular or cephalic toxoplasmosis. Either it is important to detect T. gondii from meat of food animals from retail shops or directly at slaughterhouses, which is meant for export. METHODS: The current research was done without time limitation using such terms as follows: "Toxoplasma gondii", "Meat", "Tissue cyst", "PCR", "LAMP", "Screening" and "Immunological assay" alone or in combination, in English language. The used electronic databases for searching included as follows: Pub-Med, Scopus, Google Scholar, Web of Science and Science Direct. The searches were limited to the published papers to English language. RESULTS: Sensitivity of different molecular techniques for diagnosis of Toxoplasma is real-time PCR > LAMP > conventional PCR. In addition to these DNA analysis tools, bioassay in mice and cats is considered as "gold standard" to detect T. gondii. CONCLUSION: This review article will help the readers for grasping advantages and limitations of different diagnostic tools for screening meat samples for T. gondii. This review also makes bibliography about the type of meat sample to be processed for diagnosis and different primers or sequences to be targeted for T. gondii by number of researches for its detection from meat or tissue sample using DNA amplification techniques.