Identification of immune biomarkers in recent active pulmonary tuberculosis

Tuberculosis (TB) has remained an unsolved problem and a major public health issue, particularly in developing countries. Pakistan is one of the countries with the highest tuberculosis infection rates globally. However, methods or biomarkers to detect early signs of TB infection are limited. Here, we characterized the mRNA profiles of immune responses in unstimulated Peripheral blood mononuclear cells obtained from treatment naïve patients with early signs of active pulmonary tuberculosis without previous history of clinical TB. We identified a unique mRNA profile in active TB compared to uninfected controls, including cytokines such as IL-27, IL-15, IL-2RA, IL-24, and TGFβ, transcription factors such as STAT1 and NFATC1 and immune markers/receptors such as TLR4, IRF1, CD80, CD28, and PTGDR2 from an overall 84 different transcripts analyzed. Among 12 significant differentially expressed transcripts, we identified five gene signatures which included three upregulated IL-27, STAT1, TLR4 and two downregulated IL-24 and CD80 that best discriminate between active pulmonary TB and uninfected controls with AUC ranging from 0.9 to 1. Our data identified a molecular immune signature associated with the early stages of active pulmonary tuberculosis and it could be further investigated as a potential biomarker of pulmonary T

Evaluation of the training program to train HIV treatment center staff in Pakistan

Introduction In Pakistan, HIV training programs, especially for health professionals working in HIV treatment centers, are limited. Consequently, there is little data about HIV awareness among physicians and allied health workers and how it may affect their care for people living with HIV (PLWH). Recently, the Global Fund to Fight AIDS, Tuberculosis, and Malaria (GFATM) grant Principal Recipient UNDP engaged an NGO experienced in HIV/AIDS training, on a competitive basis, to develop a training manual and conduct training of all categories of HIV treatment centers staff. The goal of this study was to assess the training program\u27s influence on trainees\u27 (both physicians and allied health staff) knowledge and abilities and describe its major lessons. Methodology This was a one-group pre-post test study, carried out between January 17 and February 22, 2023. The study was carried out in three phases. In the first phase, a team of experts developed an antiretroviral treatment (ART) training manual. In the second phase, 9- and three-day training workshops were conducted in six different cities of Pakistan, which were attended by physicians and allied health staff working in different HIV treatment centers across Pakistan. The workshops had plenary lectures, discussions, role plays, video cases, and case studies. In the third phase, a quiz, comprising multiple/best choice questions (MCQs/BCQs) and true and false questions, was administered before (pre) and after the workshop (post) to assess the impact of these training sessions in enhancing the level of HIV knowledge, especially related to ART. The workshop was attended by a total of 256 health workers from different cities in Pakistan. The participants had backgrounds in medical science, psychology, laboratory science, nursing, and computer science. Pre-and post-test responses were statistically analyzed to determine the impact of the training program on participant\u27s knowledge. For this, the Shapiro-Wilk test was applied to test data normality, followed by the application of paired t-test or Wilcoxon Signed Rank Test for normally and non-normally distributed data, respectively. Finally, a chi-square test was applied to examine the significant (p\u3c0.05) association between training workshops and improvement in the participant\u27s level of understanding of HIV. In all statistical tests, p\u3c0.05 was considered significant. Results The results from our study showed that before the training session, both physicians and allied staff possessed limited knowledge about HIV-related domains. After the workshops, participants from all cities demonstrated a uniform enhancement of knowledge related to different HIV-related domains, evident from the improvement in post-test scores compared to pre-test scores (p\u3c0.0001). The chi-square test showed a significant association between training workshops and improvement in the participant\u27s level of understanding about HIV (p-values for BCQ, MCQ, and true and false: 0.001, 0.0047, and 0.0024, respectively). Conclusions Pre- and post-test evaluation provides an objective, data-driven method for measuring the impact of educational interventions in improving healthcare workers\u27 awareness about HIV. The results emphasize the role of continuous workshops and training programs in enhancing the knowledge and understanding of healthcare and allied health workers regarding HIV

Expression profile of KIR3DS1/KIR3DL1 receptors in association with immunological responses in TB, HIV and HIV/TB infected patients

Several studies investigated KIR3DS1 and KIR3DL1 in the context of various infections. However, none of the studies were performed on KIR3DS1/L1 in association with IFN-ɣ/IL-10 in TB, HIV-1, and their confections. We aimed to evaluate KIR3DS1/KIR3DL1 expression in association with IFNɣ/IL-10 in HIV-1 and TB mono-infections and HIV-1/TB confection and compared with uninfected controls using RTq PCR. We also performed correlation analysis between KIR3DS1, KIR3DL1, IFN-ɣ and IL-10 in the respective cohorts. The overall expression of KIR3DS1 was found to be downregulated in all groups, whereas in HIV-1 and HIV-1/TB, the frequency of KIR3DS1(+) expression was significantly (p \u3c 0.05) associated with undetected HIV-1 viral load. However, expression of KIR3DL1 was found to be significantly (p \u3c 0.05) upregulated in HIV-1 only. In addition, IFNɣ expression was significantly (p \u3c 0.05) decreased in TB, whereas in HIV-1/TB, IFNɣ expression was significantly (p \u3c 0.05) increased. In contrast, IL-10 expression was significantly (p \u3c 0.05) increased in HIV-1 and HIV-1/TB but not in TB. Also, we found significant positive correlation (p \u3c 0.05, r = 0.61) between KIR3DL1 and IFNɣ expression in TB and negative correlation (p \u3c 0.05, r = - 0.62) between KIR3DS1 and IL-10 in HIV-1/TB. In conclusion, we suggest that expression of KIR3DS1/L1 is associated with IFNɣ/IL-10 responses and it is involved in modulating disease severity in HIV-1 and TB infection

Index testing among HIV-positive patients is feasible and successful in identifying additional HIV infections in Pakistan

Objective: To assess the feasibility of index testing approach to test the parents and siblings of human immunodeficiency virus (HIV)-positive children, and identify HIV-positive individuals. Methods: The study was conducted at Ratodero, Pakistan. Study participants were enrolled from September 2019 to February 2020.  The list of HIV-positive persons was provided by the Provincial Acquired Immunodeficiency Syndrome (AIDS) Control Programme. Families of 706 HIV-positive persons were approached and all of them agreed to participate. The first test was performed by trained outreach workers. All those with a reactive first test were transported to the nearest health facility for further testing and confirmation. Mothers and siblings were tested at home while additional visits were carried out to reach the fathers.  Results: A total of 1766 persons were tested through HIV index testing. Biological siblings accounted for 81% of the contacts. We were able to test 413/463 (89.2%) mothers, 232/413 (56.2%) fathers and 1121/1392 (80.5%) siblings. Out of these, 7 mothers (1.7%) and 22 siblings (2.0%) were confirmed to be HIV-positive, while no one was found to be positive among the fathers. The overall HIV prevalence was 1.6% (29/1766). All HIV-positive persons were guided for treatment and care.  Conclusion: Results indicate that index-testing approach is feasible in Pakistan to expand HIV testing services through home visits.  Key words: Human immunodeficiency virus, HIV, HIV testing, Pakistan, Index testing, HIV outbreak, Prevalence

Primary headache disorders in the adult general population of Pakistan - a cross sectional nationwide prevalence survey

Background The large geographical gaps in our knowledge of the prevalence and burden of headache disorders include almost all of Eastern Mediterranean Region (EMR). We report a nationwide population-based study in Pakistan, an EMR country with the sixth largest population in the world, conducted as a project within the Global Campaign against Headache. Methods We surveyed six locations from the four provinces of Pakistan: Punjab, Sindh, Khyber Pakhtunkhwa and Baluchistan. We randomly selected and visited rural and urban households in each. One adult member (18–65 years) of each household, also randomly selected, was interviewed by a trained non-medical interviewer from the same location using a previously-validated structured questionnaire translated into Urdu, the national language. We estimated 1-year prevalences of the headache disorders of public-health importance and examined their associations with demographic variables using multivariate analysis. Results There were 4223 participants (mean age 34.4 ± 11.0 years; male 1957 [46.3%], female 2266 [53.7%]; urban 1443 [34.2%], rural 2780 [65.8%]). Participation proportion was 89.5%. Headache in the previous year was reported by 3233 (76.6% [95% CI: 75.3–77.8%]). The age- and gender-adjusted 1-year prevalence of migraine was 22.5% [21.2–23.8%] (male 18.0% [16.8–19.2%], female 26.9% [25.6–28.2%]), of tension-type headache (TTH) 44.6% [43.1–46.1%] (male 51.2% [49.7–52.7%], female 37.9% [36.4–39.4%]), of probable medication-overuse headache 0.7% [0.5–1.0%] (male 0.7% [0.5–1.0%], female 0.8% [0.5–1.1%]) and of other headache on ≥15 days/month 7.4% [6.6–8.2%] (male 4.4% [3.8–5.0%], female 10.4% [9.5–11.3%]). Migraine was more prevalent in females by a factor of 3:2 although this association barely survived (P = 0.039) after correcting for other factors. TTH was more prevalent in males by about 4:3 (P = 0.026). All headache and migraine were age-related, peaking in the age group 40–49 years; TTH peaked a decade earlier. Higher education (P = 0.004) and income (P = 0.001) were negatively associated with prevalence of migraine. Conclusion With three quarters of its population affected, headache disorders must be on the public-health agenda of Pakistan. Worldwide, these disorders are the third leading cause of disability; information from specific enquiry into the burden attributable to headache disorders in this country is needed to inform health policy and priority-setting, and will be reported soon. Keyword