Background: The contaminated contact lens provides Pseudomonas aeruginosa an ideal site for attachment and biofilm production. Continuous contact of the eye to the biofilm-infested lens can lead to serious ocular diseases, such as keratitis (corneal ulcers). The biofilms also prevent effective penetration of the antibiotics, which increase the chances of antibiotic resistance. Methods: For this study, 22 Pseudomonas aeruginosa isolates were obtained from 36 contact lenses and 14 contact lens protective fluid samples. These isolates were tested against eight commonly used antibiotics using Kirby-Bauer disk diffusion method. The biofilm forming potential of these isolates was also evaluated using various qualitative and quantitative techniques. Finally, a relationship between biofilm formation and antibiotic resistance was also examined. Results:The isolates of Pseudomonas aeruginosa tested were found resistant to most of the antibiotics tested. Qualitative and quantitative biofilm analysis revealed that most of the isolates exhibited strong biofilm production. The biofilm production was significantly higher in isolates that were multi-drug resistant (p < 0.0001).

Background: In April 2019, an HIV screening camp for all ages was established in response to a report of an unusually large number of paediatric HIV diagnoses in Larkana, Pakistan. A cross-sectional study to understand the clinical profile of children who registered for HIV care was performed. Methods: Age-and sex-stratified HIV prevalence among individuals screened was estimated. Data on children who registered for HIV care including clinical history, HIV disease stage, Hepatitis B and C status and CD4 count was abstracted from clinical records and analysed using percentages and chi square tests. Findings: Between April-July 2019, 31,239 individuals underwent HIV testing of whom 930 tested HIV-positive. Of these, 763 (82•0%) were aged <16 years. Estimated HIV prevalence was 3.0% overall; 7.4% (283/3803) in children aged 0-2 years, 5.9% (321/5412) in children aged 3-5 years, and 1•3% (148/11251) in adults aged 16-49 years. Of the 591 children who registered for HIV care, 80.9% (478/591) were ≤5 years, 64•1% (379/591) were male and 53•4% (315/590) had a weight-forage Z-score <-3. Hepatitis B surface antigen and Hepatitis C antibody positivity was 8•4% (48/574) and 2•6% (15/574) respectively. Of children whose mothers tested for HIV, only 39/371 (10•5%) had HIV-positive mothers. Most children (89•2%, 404/453) reported multiple previous injections and 40/453 (8•8%) reported blood transfusions. Interpretation: This is an unprecedented HIV outbreak among children in Pakistan: a 54% increase in paediatric HIV diagnoses over the past 13 years. Epidemiological and molecular studies to understand the full extent of the outbreak and its drivers are needed to guide HIV control strategies.

Background We investigated the genome diversity of SARS-CoV-2 associated with the early COVID-19 period to investigate evolution of the virus in Pakistan. Materials and methods We studied ninety SARS-CoV-2 strains isolated between March and October 2020. Whole genome sequences from our laboratory and available genomes were used to investigate phylogeny, genetic variantion and mutation rates of SARS-CoV-2 strains in Pakistan. Site specific entropy analysis compared mutation rates between strains isolated before and after June 2020. Results In March, strains belonging to L, S, V and GH clades were observed but by October, only L and GH strains were present. The highest diversity of clades was present in Sindh and Islamabad Capital Territory and the least in Punjab province. Initial introductions of SARS-CoV-2 GH (B.1.255, B.1) and S (A) clades were associated with overseas travelers. Additionally, GH (B.1.255, B.1, B.1.160, B.1.36), L (B, B.6, B.4), V (B.4) and S (A) clades were transmitted locally. SARS-CoV-2 genomes clustered with global strains except for ten which matched Pakistani isolates. RNA substitution rates were estimated at 5.86 x10−4. The most frequent mutations were 5’ UTR 241C > T, Spike glycoprotein D614G, RNA dependent RNA polymerase (RdRp) P4715L and Orf3a Q57H. Strains up until June 2020 exhibited an overall higher mean and site-specific entropy as compared with sequences after June. Relative entropy was higher across GH as compared with GR and L clades. More sites were under selection pressure in GH strains but this was not significant for any particular site. Conclusions The higher entropy and diversity observed in early pandemic as compared with later strains suggests increasing stability of the genomes in subsequent COVID-19 waves. This would likely lead to the selection of site-specific changes that are advantageous to the virus, as has been currently observed through the pandemic.

Every year there are > 33 million cases of Respiratory Syncytial Virus (RSV)-related respiratory infection in children under the age of five, making RSV the leading cause of lower respiratory tract infection (LRTI) in infants. RSV is a global infection, but 99% of related mortality is in low/middle-income countries. Unbelievably, 62 years after its identification, there remains no effective treatment nor vaccine for this deadly virus, leaving infants, elderly and immunocompromised patients at high risk. The success of all pathogens depends on their ability to evade and modulate the host immune response. RSV has a complex and intricate relationship with our immune systems, but a clearer understanding of these interactions is essential in the development of effective medicines. Therefore, in a bid to update and focus our research community's understanding of RSV's interaction with immune defences, this review aims to discuss how our current knowledgebase could be used to combat this global viral threat.