COVID-19 pandemic impacted internet use and anxiety among general public during COVID-19 pandemic in Pakistan.

Objective: To determine the association between internet use and anxiety among people during the coronavirus disease- 2019 pandemic. METHODS: The cross-sectional study was conducted across Pakistan from January 14 to February 21, 2021, which was the active phase of the coronavirus disease-2019 pandemic in Pakistan. The participants were aged at least 13 years having internet access regardless of gender or their location across Pakistan. The anonymous web-based survey was conducted using a questionnaire generated on Google Forms and disseminated through various social media platforms and WhatsApp groups. Anxiety symptoms were screened using the Depression, Anxiety and Stress Scale-21, while the Young Internet Addiction Test was used to evaluate symptoms of internet addiction. Data was analysed using STATA 16. RESULTS: Of the 1,145 subjects, 686(60%) were females and 459(40%) were males. A total of 257(22.5%) participants were found to have extremely severe anxiety and internet usage pattern was significantly associated with the level of anxiety (p<0.05). Age, gender, social class and marital status were not significantly different (p>0.05), while family income and area of living were significantly different (p<0.05) in terms of anxiety levels. The odd of addictive internet use was 10.2 (95% confidence interval: 5.7-18.5) times greater in extreme anxiety individuals compared to individuals having no anxiety after controlling for other sociodemographic, health-related, behavioural and environmental factors during the pandemic. Conclusion: A significant association of anxiety was found with internet addiction during the coronavirus disease-2019 pandemic.

Mental health of caregivers of children with medical complexity: group-based trajectory modelling.

OBJECTIVE: To describe the mental health trajectories of caregivers of children with medical complexity (CMC) and explore child characteristics associated with below-average caregiver mental health. DESIGN: A secondary analysis of prospectively collected data from 123 caregivers of children aged <16 years with medical complexity from a multicentre randomised trial conducted from December 2016 to June 2021. MAIN OUTCOME MEASURE: The Patient-Reported Outcomes Measurement Information System Global Mental Health Scale was used to measure caregivers' self-reported mental health well-being. Group-based trajectory analysis was used to identify clusters of caregivers with similar changes in mental health across 24 months. Logistic regression was used to identify child-related predictors of mental health among caregivers. RESULTS: A final model with three distinct groups was selected, corresponding to caregivers with average (n=39), moderately below-average (n=65) and severely below-average (n=19) mental health scores, all with stable trajectories and high posterior probabilities (>90%). Moderately and severely below-average caregiver mental health groups, merged into one group, were associated with a greater number of child medical technology devices (adjusted OR (aOR) 1.44, 95% CI 1.01 to 2.04), gross motor difficulties (aOR 3.51, 95% CI 1.02 to 12.05) and worse child emotional (aOR 0.93, 95% CI 0.87 to 0.99) and psychological well-being (aOR 0.93, 95% CI 0.88 to 0.99). CONCLUSION: Most caregivers of CMC reported persistently below-average mental health. The intensity of caregiving, as indicated by medical technology and child functional needs, is a potential risk factor for below-average caregiver mental health. Future design and evaluation of interventions focused on support for caregivers of CMC are warranted.

Adverse perinatal events and maternal interpregnancy weight change: A population-based observational study.

OBJECTIVE: Mothers whose newborn experiences adversity may neglect their own health to care for their affected infant or following a perinatal death. Weight gain after pregnancy is one measure of maternal self-care. We measured interpregnancy weight gain among women whose child had an adverse perinatal event. METHODS: This population-based observational study included 192 154 primigravid women with two consecutive singleton births in Ontario, Canada. Outcomes included net weight gain, and adjusted odds ratios (aOR) of moving to a higher body mass index (BMI) category between pregnancies, comparing women whose child did versus did not experience either a perinatal death, prematurity, severe neonatal morbidity, major congenital anomaly, or severe neurologic impairment. RESULTS: Perinatal death was associated with a +3.5 kg (95% confidence interval [CI]: 2.1-4.9) net higher maternal weight gain in the subsequent pregnancy. Relative to term births, preterm birth <32 weeks (+3.2 kg, 95% CI: 1.9-4.6), 32-33 weeks (+1.8 kg, 95% CI: 0.7-2.8) and 34-36 weeks (+0.9 kg, 95% CI: 0.6-1.3) were associated with higher net weight gain. Having an infant with severe neonatal morbidity was associated with a +1.2 kg (95% CI: 0.3-2.1) weight gain. Likewise, the aOR of moving to a higher BMI category was 1.27 (95% CI, 1.14-1.42) following a perinatal death, 1.21 (95% CI: 1.04-1.41) after a preterm birth <32 weeks, and 1.11 (95% CI: 1.02-1.22) with severe neonatal morbidity. CONCLUSION: Greater interpregnancy weight gain, and movement to a higher BMI category, are each more likely in a woman whose first-born was affected by certain major adverse perinatal events.

Risk factors for acute diarrhoea in children between 0 and 23 months of age in a peri-urban district of Pakistan: a matched case-control study.

BACKGROUND: Diarrhoea is a leading cause of preventable childhood morbidity and mortality worldwide. Unfortunately, Pakistan has the third-highest burden of diarrhoea-related deaths in children <5 y of age. Therefore we aimed to evaluate factors associated with diarrhoea among Pakistani children. METHODS: A retrospective 1:2 matched case-control study nested in a baseline cross-sectional survey was conducted from October to December 2018 in Taluka Kotri, a two-thirds urban locality in the Jamshoro district. Children between the ages of 0 and 23 months with a history of diarrhoea in the 2 weeks preceding the survey were labelled as cases. Age-matched controls were children without symptoms of diarrhoea. Univariate and multivariable conditional logistic regression was performed to identify diarrhoea-related factors. RESULTS: A total of 1558 cases were matched with 3116 controls. Factors significantly associated with lower odds of diarrhoea in the multivariate analysis included increasing maternal age (odds ratio [OR] 0.78 [95% confidence interval {CI} 0.67 to 0.90]), breastfeeding (OR 0.77 [95% CI 0.66 to 0.90]), higher paternal education (OR 0.79 [95% CI 0.65 to 0.97]) and belonging to the rich (OR 0.66 [95% CI 0.54 to 0.80]) and richest quintiles (OR 0.54 [95% CI 0.44 to 0.66]). CONCLUSIONS: This study identifies risk factors associated with diarrhoea in children <23 months of age, including younger maternal age, higher paternal education, not breastfeeding and poverty, which has implications for developing preventive programs and strategies that target populations with a higher risk of diarrhoea.

Severe Respiratory Disease Among Children With and Without Medical Complexity During the COVID-19 Pandemic.

Importance: Severe respiratory disease declined during the COVID-19 pandemic, partially due to decreased circulation of respiratory pathogens. However, the outcomes of children with higher risk have not been described using population-based data. Objective: To compare respiratory-related hospitalizations, intensive care unit (ICU) admissions, and mortality during the pandemic vs prepandemic, among children with medical complexity (CMC) and without medical complexity (non-CMC). Design, Setting, and Participants: This population-based repeated cross-sectional study used Canadian health administrative data of children aged younger than 18 years in community and pediatric hospitals during a pandemic period (April 1, 2020, to February 28, 2022) compared with a 3-year prepandemic period (April 1, 2017, to March 31, 2020). The pandemic period was analyzed separately for year 1 (April 1, 2020, to March 31, 2021) and year 2 (April 1, 2021, to February 28, 2022). Statistical analysis was performed from October 2022 to April 2023. Main Outcomes and Measures: Respiratory-related hospitalizations, ICU admissions, and mortality before and during the pandemic among CMC and non-CMC. Results: A total of 139 078 respiratory hospitalizations (29 461 respiratory hospitalizations for CMC and 109 617 for non-CMC) occurred during the study period. Among CMC, there were fewer respiratory hospitalizations in both 2020 (rate ratio [RR], 0.44 [95% CI, 0.42-0.46]) and 2021 (RR, 0.55 [95% CI, 0.51-0.62]) compared with the prepandemic period. Among non-CMC, there was an even larger relative reduction in respiratory hospitalizations in 2020 (RR, 0.18 [95% CI, 0.17-0.19]) and a similar reduction in 2021 (RR, 0.55 [95% CI, 0.54-0.56]), compared with the prepandemic period. Reductions in ICU admissions for respiratory illness followed a similar pattern for CMC (2020: RR, 0.56 [95% CI, 0.53-0.59]; 2021: RR, 0.66 [95% CI, 0.63-0.70]) and non-CMC (2020: RR, 0.22 [95% CI, 0.20-0.24]; RR, 0.65 [95% CI, 0.61-0.69]). In-hospital mortality for these conditions decreased among CMC in both 2020 (RR, 0.63 [95% CI, 0.51-0.77]) and 2021 (RR, 0.72 [95% CI, 0.59-0.87]). Conclusions and Relevance: This cross-sectional study found a substantial decrease in severe respiratory disease resulting in hospitalizations, ICU admissions, and mortality during the first 2 years of the pandemic compared with the 3 prepandemic years. These findings suggest that future evaluations of the effect of public health interventions aimed at reducing circulating respiratory pathogens during nonpandemic periods of increased respiratory illness may be warranted.

Burden and factors associated with perceived stress amidst COVID-19: a population web-based study in Pakistan.

OBJECTIVE: This study aims to determine the burden and factors associated with perceived stress in the Pakistani population amidst the COVID-19 pandemic. SETTING: A web-based cross-sectional survey was conducted from April to August 2020. POPULATION: This survey was broadcasted on the web using a Google form link and 1654 Pakistani residents had completed this survey. Individuals belonging to any province, city, village, or district of Pakistan irrespective of any age, having internet access and a link of Google form, with English/Urdu competency, consent to participate, and currently residing in Pakistan were eligible to participate. OUTCOME MEASURE: Perceived stress was measured using a validated tool of perceived stress scale-10. Multiple ordinal regression was used, and an adjusted OR along with a 95% CI are reported. RESULTS: The mean score of perceived stress was 19.32 (SD ±6.67). Most of the participants screened positive for moderate (69%) and high levels (14%) of stress, respectively. The odds of high-perceived stress among severely anxious participants were 44.67 (95% CI: 21.33 to 93.53) times than participants with no/minimal generalised anxiety during the complete lockdown. However, the odds of high levels of perceived stress among moderately anxious respondents were 15.79 (95% CI: 10.19 to 24.28) times compared with participants with no/minimal anxiety during the smart lockdown. CONCLUSION: This study evidence that the pandemic was highly distressing for the Pakistani population causing the maximum level of perceived stress in more than half of the population. Adequate and timely interventions are needed before high-stress levels culminate into psychological disorders.

CMV and EBV Co-Infection in HIV-Infected Children: Infection Rates and Analysis of Differential Expression of Cytokines in HIV Mono- and HIV-CMV-EBV Co-Infected Groups.

(1) Background: CMV and EBV co-infections can affect the HIV disease progression by modulating the immune system. The disease dynamics can differ in HIV-positive adults and children. In Pakistan, HIV is rapidly expanding, especially in children; however, the prevalence of CMV and EBV co-infection and the effect on immune modulation in HIV-positive children are not known. This study aimed to bridge this gap by estimating the rate of active CMV and EBV co-infection in HIV-positive children, followed by the analysis of differential expression of cytokines in HIV mono- and HIV/CMV/EBV co-infected children. (2) Methods: DNA samples from 319 HIV-positive children, previously recruited as part of a study to investigate the HIV outbreak in Larkana, Pakistan, in 2019, were screened for CMV and EBV through qPCR. Subsequently, differences in HIV viral loads and CD4 counts were analyzed between the HIV mono- and HIV/CMV/EBV co-infected groups. The RNA samples were used to determine the differential expression of both pro- and anti-inflammatory cytokines in the mono- and co-infected groups using RT-qPCR, while unpaired T-test and Pearson correlation test were applied to, respectively, analyze the differential cytokine expression and correlation between cytokine in the two groups. (3) Results: Of 319 samples, the rate of active EBV and CMV co-infection in HIV-positive children was observed in 79.9% and 38.9%, respectively. A significant difference was observed in HIV viral load between HIV mono- and co-infected groups. IFN-γ expression was found to be lower in the HIV mono-infected group, while higher in all other three co-infected groups. Meanwhile, mRNA expression of TGF-ß1 was found to be lower in HIV mono- and HIV-CMV-EBV co-infected groups, while higher in HIV-CMV and HIV-EBV co-infected groups. IFN-γ and IL-2 exhibited a significant positive correlation in all except HIV-CMV co-infected group. (4) Conclusions: The study suggests that the presence of EBV/CMV co-infection can affect the HIV viral loads and expression of certain cytokines (IFN-γ and TGF-ß1), which may affect the HIV disease dynamics in infected children.

Effect of frequency of parent to child maltreatment on symptoms of anxiety disorder in Pakistani adolescents: A community based cohort study.

OBJECTIVE: To investigate the association between Parent to Child Maltreatment and the occurrence of anxiety disorder symptoms among Pakistani adolescents under 18 years. METHOD: A prospective study was conducted to examine the effect of frequency of Parent to child maltreatment (PTC) on the occurrence of anxiety disorders in a community setting. A sample of 800 adolescents who reported none to rare, occasional, and frequent were followed for the occurrence of anxiety disorder symptoms over two years. PTC was evaluated using ICAST-C (International child abuse screening tool). Anxiety disorder symptoms were evaluated using the SCARED (Screen for children anxiety-related disorders) tool. Risk ratios were estimated using Cox Proportional Algorithm. RESULTS: The risk of occurrence of anxiety disorder symptoms in adolescents who have exposure to frequent PTC is 3.8 times (2.9, 4.9) as compared to those with none to rare exposure to PTC. Among adolescents suffering frequent PTC whose, mothers also reported a history of domestic violence, the risk of anxiety disorder symptoms is 3 times (2.2, 4.1) higher. Female gender (RR 1.5; 1.2-1.9), stressful home environment (RR 1.4; 1.1-1.8), and parental substance abuse (RR 1.6; 1.2-2.0) are significant predictors of anxiety disorder symptoms. CONCLUSION: Frequent PTC is significantly associated with the occurrence of anxiety disorder symptoms among Pakistani adolescents. Given these findings, there is a dire need to develop specific interventions to address anxiety disorder symptoms among adolescents and to develop programs to prevent parent to child maltreatment.

Unassigned Complex Unique Recombinant Forms Related to CRF36_cpx in Children Identified in an HIV-1 Outbreak in Pakistan.

In 2019, an outbreak of HIV infection predominantly affecting children occurred in Larkana district, Pakistan. This is the largest outbreak ever reported in this age group in Pakistan. In this study, we report two HIV-1 unique recombinant forms identified during the outbreak. Blood samples were collected from HIV-positive children as part of a case-control study to investigate the outbreak. The pol gene was sequenced and used to detect HIV subtype/recombinant forms using subtype, recombination, and phylogenetic analyses. Drug resistance mutation (DRM) analysis was performed to characterize the DRMs in each sequence. We observed the emergence of two unassigned unique recombinant forms related to CRF36_cpx in 15 individuals of the 344 samples collected. Genotype analysis revealed the presence of multiple DRMs associated with resistance to reverse transcriptase inhibitors. The discovery of these unassigned unique recombinant forms in our population highlights the need for comprehensive molecular epidemiological studies to fully understand the distribution and drug resistance patterns to aid control efforts.

Phylogenetic and drug- and vaccine-resistance profiles of Hepatitis B Virus among children with HIV co-infection in Pakistan.

INTRODUCTION: HIV-1 and hepatitis B virus (HBV) share common routes of transmission and therefore co-infection is common. In 2019, an HIV-1 outbreak that resulted in >1000 children being infected, predominantly through nosocomial transmission, occurred in Sindh, Pakistan. We conducted a phylogenetic and drug resistance analysis of the HBV Reverse Transcriptase (RT) gene in children with HIV-1 and HBV co-infection. METHODOLOGY: Blood samples were collected from 321 children with HIV who were recruited as part of a study to investigate the HIV-1 outbreak. All samples were tested for HBV surface antigen (HBsAg) using an ELISA assay, and positive samples were used to amplify and sequence the HBV RT gene. The phylogenetic relationship between sequences was analyzed, and drug- and vaccine- resistance mutations in the RT gene were explored. RESULTS: Of 321 samples, 23% (n = 75) were positive for HBsAg on ELISA. Phylogenetic analysis of the sequences revealed that 63.5% of HBV sequences were sub-genotype D1, while the rest were sub-genotype D2. Cluster analysis revealed grouping of sub-genotype D1 sequences exclusively with Pakistani sequences, while clustering of sub-genotypes D2 predominantly with global sequences. The 236Y mutation associated with resistance to tenofovir was observed in 2.8% of HBV sequences. Additionally, seven vaccine escape mutations were observed, the most common being 128 V. CONCLUSION: Our study suggests ongoing transmission of HBV D1 and D2 sub-genotypes in the HIV-1 co-infected population, likely nosocomially, given common routes of HVB and HIV-1 transmission. The prevalence of major HBV drug- and vaccine-resistant mutations remains low. Surveillance for further transmissions and the possible emergence of major drug- or vaccine-resistant variants is required.

Exploring terminology for puerperal sepsis and its symptoms in urban Karachi, Pakistan to improve communication, care-seeking, and illness recognition.

Puerperal sepsis is an important cause of maternal morbidity and mortality in developing countries. Awareness of local terminology for its signs and symptoms may improve communication about this illness, what actions to take when symptoms appear, timely care seeking, and clinical outcomes. This formative research aimed to improve recognition and management of postpartum sepsis in Pakistan by eliciting local terms used for postpartum illnesses and symptoms. We conducted 32 in-depth interviews with recently delivered women, their relatives, traditional birth attendants, and health care providers to explore postpartum experiences. Terms for symptoms and illness are used interchangeably (i.e. bukhar, the Urdu word for fever), many variations exist for the same term, and gradations of severity for each term as not associated with different types of illnesses. The lack of a designated term for postpartum sepsis in Urdu delays care-seeking and proper diagnosis, particularly at the community level. Ideally, a common lexicon for symptoms and postpartum sepsis would be developed but this may not be feasible or appropriate given the nature of the Urdu language and local understandings of postpartum illness. These insights can inform how we approach educational campaigns, the development of clinical algorithms that focus on symptoms, and counselling protocols.

Demographic and Psychosocial Factors associated with Internet Addiction among the Pakistani Population during COVID-19: A Web-Based Survey.

This study aims to determine the burden of COVID19-induced internet addiction and related psychosocial factors among the Pakistani sample. A web-based cross-sectional survey was conducted from January to March 2021. An analytical cross-sectional survey was broadcast on the internet via a Google form completed by 1145 Pakistanis. Individuals aged 13 years and above with competency to comprehend English or Urdu language, currently residing in any province of Pakistan, having access to the questionnaire, and willing to participate were eligible to participate. Overseas Pakistanis were excluded from the study. The outcome is COVID19-induced internet addiction was measured using the validated tool Young's Internet Addiction Test (IAT). In addition, symptoms of depression, anxiety, stress, and other psychosocial factors were assessed using the validated tool Depression, Anxiety, and Stress Scale-21 (DASS-21). Adjusted odds ratios with a 95% confidence interval were reported using multinomial logistic regression. Most participants were females and youth (between 20 and 24 years). The prevalence of problematic internet users (PIU) and addictive internet users (AIU) was 27.3% and 11.3%, respectively. The odds of extremely severe anxiety among AIU were approximately three times (Adj OR: 2.6 (1.1-7.1), followed by the odds of having extremely severe stress being about five times higher among AIU (Adj OR: 5.4 (1.6-17.6)) as compared to normal internet users (NIU). Amid COVID-19, the burden of internet addiction has surged among the Pakistani populace. This study identified that gender, marital status, depression, stress, anxiety, work situation, and mood changes during the COVID-19 pandemic are significantly correlated with problematic and addictive internet use. Preventative measures against the addictive use of the internet are needed to avoid or mitigate any serious mental health problems.

The effect of a culturally sensitive mental well-being module on Pakistani nursing students' knowledge and beliefs regarding their own mental health and illness.

This study examined the effect of a mental well-being module on undergraduate nursing students' knowledge and beliefs regarding causes and treatment of mental health and illness, and explored the relationship between these variables and depression and anxiety. The one group design incorporated pre-intervention, intervention and post-intervention phases, in a consecutive sample of 246 students in a 4-year BScN programme in Pakistan. A culturally sensitive, interactive mental well-being module of four hours length was delivered to three groups of approximately 40 students per group. Repeated measures ANOVA was performed to assess the difference in the mean knowledge scores. Paired t-tests investigated between-group differences on the factor scores. A total of 112 students screened positive for anxiety and depression symptoms. Repeated measures ANOVA showed a significant difference in the mean knowledge scores regarding mental illness (P < 0.001). Factor analysis resulted in four factors for the aetiology items. Paired t-test showed significant differences (P < 0.001) between psycho-social and environmental factors, supernatural and religious beliefs, and neuro-genetics. For treatment, significant differences (P < 0.001) were found between all factors - Professional and Help from others, Religiosity and Socialization, and Alternative and Medical Treatment. There was a significant and positive change in students' mental health knowledge, beliefs and mental health-seeking behaviour and diminished stigmatized beliefs. Based on the findings, it is highly recommended to incorporate the mental well-being program in the curriculum.

The role of parent-to-child maltreatment in the pathway of self-reported depressive symptoms in Pakistani adolescents.

BACKGROUND: Parent-to-child maltreatment has been demonstrated to drastically affect a child's mental well-being and plays a significant role in developing depressive symptoms. However, little is established about the effect of frequency of parent-to-child maltreatment on the development of depressive symptoms among Pakistani adolescents. METHODS: A longitudinal prospective study was conducted, from 2015 to 2017, with 800 adolescents aged 11-17 years old recruited from 32 systematically selected urban and peri-urban areas of Karachi. First, these adolescents were screened for parent-to-child maltreatment in 2015 in a cross-sectional survey. Children with diagnosed psychiatric conditions were excluded from the study. In the second phase, these individuals were followed for 2 years to investigate the symptoms of depressive disorder using a validated tool, "CES-D (Center for Epidemiological Studies) Depression scale." The Cox proportional algorithm was used to examine the relationship between the frequency of parent-to-child maltreatment and depressive symptoms. RESULTS: Approximately 11% of frequently, 9% of occasionally parent-to-child, and 7%of negligibly maltreated adolescents reported depressive symptoms over 2 years. The other significant predictors of depressive symptoms were no formal education of the child (RR: 3.15, 95% CI: 1.35-7.34), presence of stressful home environment (RR: 2.19, 95% CI: 1.22-3.94), and having both uneducated parents (RR: 1.70, 95% CI: 0.90-3.21). The frequently maltreated females were found to have 4 times the higher risk compared to rarely maltreated males. In addition, frequently maltreated males were twice likely to develop depressive symptoms. CONCLUSION: The results suggested that frequent parent-to-child maltreatment occurring during childhood leads to the development of depressive symptoms later in the adolescence period. Thus, there is a dire need for interventions to raise awareness among the society on the issue of parent-to-child mistreatment to minimize later mental health consequences.

Longitudinal association between parental-to-child-Maltreatment and self-reported Generalized Anxiety Disorder symptoms in Pakistani Adolescents.

BACKGROUND: Parent-to-child maltreatment is considered one of the risk factors for Generalized Anxiety Disorder (GAD) symptoms, but this hypothesis has not been adequately tested in Pakistani settings. AIM: This study aimed to examine the association between parent-to-child maltreatment and the risk of developing GAD symptoms among adolescents. METHODS: The association of none to rare, occasionally, and frequently parent-to-child maltreatment with the incidence of GAD symptoms was investigated in a sample of 800 adolescents aged 11-17 years who were followed for two years. Parent-to-child maltreatment was assessed using ICAST-C (International child abuse screening tool). GAD Symptoms were determined by SCARED (Screen for children anxiety-related disorders). Cox Proportional Algorithm was used to estimate risk ratios. RESULTS: Among children with both uneducated parents, frequently maltreatment was associated with 7.31 (2.20-24.04) times the risk of GAD symptoms compared to none to rare maltreatment. In contrast, the risk of GAD symptoms in frequently maltreated children was 5.58 times (1.40-21.97) than negligibly maltreated children with either educated parent. CONCLUSION: The frequency of parent-to-child maltreatment is significantly associated with an increased risk of developing GAD symptoms in which parental education plays a crucial role. Parents should be imparted with the awareness of the consequences of child maltreatment. In Pakistani settings the need to have this awareness is even more necessary due to the culturally acceptable disciplinary measures used by parents.

Impact of pulse oximetry on hospital referral acceptance in children under 5 with severe pneumonia in rural Pakistan (district Jamshoro): protocol for a cluster randomised trial.

BACKGROUND: Pneumonia is a leading cause of death among children under 5 specifically in South Asia and sub-Saharan Africa. Hypoxaemia is a life-threatening complication among children under 5 with pneumonia. Hypoxaemia increases risk of mortality by 4.3 times in children with pneumonia than those without hypoxaemia. Prevalence of hypoxaemia varies with geography, altitude and severity (9%-39% Asia, 3%-10% African countries). In this protocol paper, we describe research methods for assessing impact of Lady Health Workers (LHWs) identifying hypoxaemia in children with signs of pneumonia during household visits on acceptance of hospital referral in district Jamshoro, Sindh. METHODS AND ANALYSIS: A cluster randomised controlled trial using pulse oximetry as intervention for children with severe pneumonia will be conducted in community settings. Children aged 0-59 months with signs of severe pneumonia will be recruited by LHWs during routine visits in both intervention and control arms after consent. Severe pneumonia will be defined as fast breathing and/or chest in-drawing, and, one or more danger sign and/or hypoxaemia (Sa02 <92%) in PO (intervention) group and fast breathing and/or chest in-drawing and one or more danger sign in clinical signs (control) group. Recruits in both groups will receive a stat dose of oral amoxicillin and referral to designated tertiary health facility. Analysis of variance will be used to compare baseline referral acceptance in both groups with that at end of study. ETHICS AND DISSEMINATION: Ethical approval was granted by the Ethics Review Committee of the Aga Khan University (4722-Ped-ERC-17), Karachi. Study results will be shared with relevant government and non-governmental organisations, presented at national and international research conferences and published in international peer-reviewed scientific journals. TRIAL REGISTRATION NUMBER: NCT03588377.

Risk Factors for Acute Respiratory Infections in Children Between 0 and 23 Months of Age in a Peri-Urban District in Pakistan: A Matched Case-Control Study.

Background: Acute respiratory infection (ARI) accounts for nearly 15% of all childhood mortality in South Asia, with children from rural areas at higher risk due to inaccessibility to healthcare facilities. We therefore aimed to identify risk factors associated with ARI in children under 2 years of age in rural Pakistan. Methods: A retrospective 1:2 matched case-control study was conducted between October and December 2018 in Taluka Kotri, Jamshoro District of Pakistan. Cases were identified as children between 0 and 23 months of age with a history of fever, cough, sore throat, fast breathing, difficulty breathing, or chest indrawing in the 2 weeks prior to the survey. Controls were participants without symptoms of ARI, matched based on age in months. Data analysis was conducted using STATA version 15. Univariate and multivariable conditional logistic regression analyses were used to identify factors associated with ARI, and p < 0.05 was considered statistically significant. Results: We identified 1,071 cases of ARI who were matched with 2,142 controls. Multivariable analysis revealed that female gender [odds ratio (OR) 0.78, 95% confidence interval (CI): 0.67-0.91], exclusive breastfeeding (OR 0.81, 95% CI: 0.69-0.97), and comorbidity with diarrhea (OR: 1.64, 95% CI: 1.40-1.91) were significantly associated with ARI. Conclusion: Pakistan continues to progress toward reducing childhood mortality, particularly ARI-related deaths, for which it bears a great burden. This study identifies risk factors such as the male gender, breastfeeding, and comorbidities with diarrhea, which could open grounds for further programmatic implications in targeting a multifaceted approach to reducing incidences of ARI in rural areas of the country.

Factors associated with HIV infection among children in Larkana District, Pakistan: a matched case-control study.

BACKGROUND: In April, 2019, an HIV outbreak predominantly affecting children occurred in Larkana District, Sindh, Pakistan. By December, 2019, 881 (4·0%) of 21 962 children screened for HIV had tested positive. We aimed to assess factors associated with HIV infection in this outbreak. METHODS: In this individually matched case-control study, we sampled 406 cases (individuals aged <16 years who had registered for paediatric HIV care at the HIV Treatment Centre at Shaikh Zayed Children's Hospital in Larkana City, Pakistan) and 406 controls (individuals without HIV matched by age, sex, and neighbourhood residence, recruited through doorknocking at houses adjacent to case participants). An interviewer-administered questionnaire was used to collect data on possible risk factors for HIV acquisition and a blood sample was collected from all participants for hepatitis B and hepatitis C serology. Mothers of all participants underwent HIV testing. Odds ratios were estimated using conditional logistic regression to assess factors associated with HIV infection. FINDINGS: 406 case-control pairs were recruited between July 3 and Dec 26, 2019. Five pairs were excluded (three pairs had an age mismatch and two pairs were duplicate cases) and 401 were analysed. The prevalence of hepatitis B surface antigen was 18·2% (95% CI 14·5-22·3) among cases and 5·2% (3·3-7·9) among controls, and the prevalence of hepatitis C antibodies was 6·5% (95% CI 4·3-9·4) among cases and 1·0% (0·3-2·5) among controls. 28 (7%) of 397 mothers of cases for whom we had data, and no mothers of 394 controls, were HIV positive. In the 6 months before recruitment, 226 (56%) of 401 cases and 32 (8%) of 401 controls reported having more than ten injections, and 291 (73%) cases and 78 (19%) controls had received an intravenous infusion. At least one blood transfusion was reported in 56 (14%) cases and three (1%) controls in the past 2 years. HIV infection was associated with a history of more injections and infusions (adjusted odds ratio 1·63; 95% CI 1·30-2·04, p<0·0001), blood transfusion (336·75; 23·69-4787·01, p<0·0001), surgery (399·75, 13·99-11 419·39, p=0·0005), the child's mother being HIV positive or having died (3·13, 1·20-8·20, p=0·020), and increased frequency of private clinic (p<0·0001) and government hospital visits (p<0·0001), adjusting for confounders. INTERPRETATION: The predominant mode of HIV transmission in this outbreak was parenteral, probably due to unsafe injection practices and poor blood safety practices. General practitioners across Pakistan need training and systems support in reducing injection use, and in providing safe injections and transfusions only when necessary. FUNDING: Department of Pediatrics and Child Health, the Aga Khan University, Karachi, Pakistan.

Phylogenetic and Drug-Resistance Analysis of HIV-1 Sequences From an Extensive Paediatric HIV-1 Outbreak in Larkana, Pakistan.

INTRODUCTION: In April 2019, an HIV-1 outbreak among children occurred in Larkana, Pakistan, affecting more than a thousand children. It was assumed that the outbreak originated from a single source, namely a doctor at a private health facility. In this study, we performed subtype distribution, phylogenetic and drug-resistance analysis of HIV-1 sequences from 2019 outbreak in Larkana, Pakistan. METHODS: A total of 401 blood samples were collected between April-June 2019, from children infected with HIV-1 aged 0-15 years recruited into a case-control study to investigate the risk factors for HIV-1 transmission. Partial HIV-1 pol sequences were generated from 344 blood plasma samples to determine HIV-1 subtype and drug resistance mutations (DRM). Maximum-likelihood phylogenetics based on outbreak and reference sequences was used to identify transmission clusters and assess the relationship between outbreak and key population sequences between and within the determined clusters. Bayesian analysis was employed to identify the time to the most recent common recent ancestor (tMRCA) of the main Pakistani clusters. RESULTS: The HIV-1 circulating recombinant form (CRF) 02_AG and subtype A1 were most common among the outbreak sequences. Of the treatment-naïve participants, the two most common mutations were RT: E138A (8%) and RT: K219Q (8%). Four supported clusters within the outbreak were identified, and the median tMRCAs of the Larkana outbreak sequences were estimated to 2016 for both the CRF02_AG and the subtype A1 clusters. Furthermore, outbreak sequences exhibited no phylogenetic mixing with sequences from other high-risk groups of Pakistan. CONCLUSION: The presence of multiple clusters indicated a multi-source outbreak, rather than a single source outbreak from a single health practitioner as previously suggested. The multiple introductions were likely a consequence of ongoing transmission within the high-risk groups of Larkana, and it is possible that the so-called Larkana strain was introduced into the general population through poor infection prevention control practices in healthcare settings. The study highlights the need to scale up HIV-1 prevention programmes among key population groups and improving infection prevention control in Pakistan.

Investigation of an extensive outbreak of HIV infection among children in Sindh, Pakistan: protocol for a matched case-control study.

INTRODUCTION: In April 2019, 14 children were diagnosed with HIV infection by a private healthcare provider in Larkana district, Sindh province, Pakistan. Over the next 3 months, 930 individuals were diagnosed with HIV, >80% below 16 years, the largest ever outbreak of HIV in children in Pakistan. In this protocol paper, we describe research methods for assessing likely modes of HIV transmission in this outbreak and investigate spatial and molecular epidemiology. METHODS AND ANALYSIS: A matched case-control study will be conducted with 406 cases recruited. Cases will be children aged below 16 years registered for care at the HIV treatment centre at Shaikh Zayed Children Hospital in Larkana City. Controls will be children who are HIV-uninfected (confirmed by a rapid HIV test) matched 1:1 by age (within 1 year), sex and neighbourhood. Following written informed consent from the guardian, a structured questionnaire will be administered to collect data on sociodemographic indices and exposure to risk factors for parenteral, vertical and sexual (only among those aged above 10 years) HIV transmission. A blood sample will be collected for hepatitis B and C serology (cases and controls) and HIV lineage studies (cases only). Mothers of participants will be tested for HIV to investigate the possibility of mother-to-child transmission. Conditional logistic regression will be used to investigate the association of a priori defined risk factors with HIV infection. Phylogenetic analyses will be conducted. Global positioning system coordinates of participants' addresses will be collected to investigate concordance between the genetic and spatial epidemiology. ETHICS AND DISSEMINATION: Ethical approval was granted by the Ethics Review Committee of the Aga Khan University, Karachi. Study results will be shared with Sindh and National AIDS Control Programs, relevant governmental and non-governmental organisations, presented at national and international research conferences and published in international peer-reviewed scientific journals.