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An experimental study was conducted to compare the predicted performance of a commercially available coil type liquid desiccant cooling and dehumidification system with the experimental results at part-load conditions. A mathematical model was used to predict the performance of coil type liquid desiccant system. This model was based mostly on the manufacturer's catalog data. The coil type system used in the experimental study was a system manufactured in the U.S.A. The system is commercially available as 200-10 KXCb, 10-row deep conditioner unit with a design air flow rate of 1180 1/s (2500 scfm). For regeneration of the solution, a packed type 3.0-P regenerator unit with a design air flow rate of 448.4 1/s (950 scfm) was used.

Laser surface modification is an emerging process that can produce texture on a work surface and effectively enhance surface topography while altering surface roughness. Laser surface modification is a sensitive process that depends on various laser processing parameters such as power, scanning speed, hatching distance. The significance of this work is to examine the influence of hatching distance on the surface characteristic of 2205 duplex stainless steel samples. The surface transformation and variation of the surface roughness properties of the materials were examined. The hatching distance was varied from 0.1 to 0.005 mm. Results indicate that, as the hatch spacing decreases, the overlap of laser track increases, thereby resulting in a decrease of surface roughness. Meanwhile, with the increase of hatch distance, the clear overlay tracks were transformed to irregular wavy surface. The best hatch distance parameter obtained was 100 ìm that resulted in the highest roughness of 8.45 ìm. Experimental results illustrate that, when the optimum hatch distance of 100 ìm was adopted, the polished smooth surface of 2205 duplex stainless steel with initial average roughness value of 0.19 ìm increased by 42 times of the polished surface roughness. A strong correlation between hatching distance and roughness was established in 2205 duplex stainless steel. High depth of the altered surface topography and increased roughness were linked to higher levels of hatching distance. [ABSTRACT FROM AUTHOR]

A newly developed medium-carbon carbide-free bainitic steel was fabricated for the first time utilizing the laser powder bed fusion (L-PBF) technique. Process parameters were optimized, and a high density of 99.8% was achieved. The impact of austempering heat treatment on the bainite morphology and transformation kinetics was investigated by high-resolution microstructural analysis (SEM, TEM, and EDS) and dilatometric analysis, and results were compared with conventionally produced counterparts. Faster kinetics and finer microstructures in the L-PBF specimens were found as a consequence of the as-built microstructure, characterized by fine grains and high dislocation density. However, a bimodal distribution of bainitic ferrite plate thickness (average value 60 nm and 200 nm, respectively) was found at prior melt pool boundaries resulting from carbon depletion at such sites.

Primary cardiac tumours are a rare occurrence, of which lipomas comprise approximately 8%. Although mostly asymptomatic, cardiac lipomas can lead to lethal arrhythmias and significant left ventricular outflow obstruction. We present a case of an asymptomatic left ventricular lipoma managed by surgical resection and discuss diagnostic modalities and management options. Our case exemplifies how prompt surgical resection is a reasonable and safe approach in select patients even if they are asymptomatic. Résumé: Les tumeurs cardiaques primitives sont rares; environ 8 % d’entre elles sont des lipomes. Bien qu’ils soient généralement asymptomatiques, les lipomes cardiaques peuvent entraîner des arythmies mortelles et provoquer une obstruction importante de la voie d’éjection du ventricule gauche. Nous exposons un cas de lipome du ventricule gauche asymptomatique pris en charge par résection chirurgicale et traitons des outils diagnostiques et des options thérapeutiques. Ce cas montre qu’il peut être raisonnable et sûr de procéder rapidement à une résection chirurgicale dans certains cas, même si le patient ne présente pas de symptômes.

Stress corrosion cracking (SCC) of laser powder bed fusion-fabricated 316L was studied under the variation in energy input density to emulate the existence of distinctive types of defects. Various electrochemical polarization measurements were performed in as-received polished and ground states, to elucidate the effect of defect type on corrosion and SCC behaviour in marine solution. The results revealed severe localized corrosion attack and SCC initiation for specimens with a lack of fusion pores (LOF). Moreover, the morphology of SCC was different, highlighting a more dominant effect of selective dissolution of the subgrain matrix for gas porosities and a more pronounced effect of brittle fracture at laser track boundaries for the specimens with LOF pores.

جوهر بحثنا يتمثل بطرح التساؤل:كيف وظفت القنوات التلفزيونية الفضائية شبكات التواصل الاجتماعي،لتعزيز التشاركية التفاعلية وخلق بيئة تفاعلية مع الجمهور العراقي،إزاء القضايا السياسية العراقية والإقليمية والدولية؟.وبناءً عليه،تم طرح مجموعة تساؤلات فرعية،تبحث عن آليات التوظيف،ومدى تزويد الجمهور بالمعرفة عن القضايا السياسية العراقية،و مدى إتاحة أدوات التفاعلية،وما القنوات الفضائية التي تمكن تعزيز التشاركية،والمجال الجغرافي للقضايا،وما حجم المنشورات،والتفاعلية.هذا البحث يهدف للتقصي عن حجم التفاعلية "التعليقات، المشاهدات، الاعجابات، المشاركات" ومعرفة مقدار المنشورات بقناة "الشرقية" على الشبكات "فيس بوك، توتير، يوتيوب إزاء القضايا السياسية العراقية، وصيغ المنشورات وطبيعة الاتجاهات. واستخدام المنهج المسحي، وطريقة تحليل المضمون، لفحص مضمون معلومات محددة، لتصوير الواقع الفعلي لحدود التوظيف.

Surface quality and dimensional tolerances of the selective laser melting (SLM) process are not good enough for many industrial applications and grinding as a common finishing process introduces many surface modifications. Investigation on the effect of grinding induced surface residual stress (RS) on early stages of stress corrosion cracking (SCC) of SLM manufactured 316L austenitic stainless steel was conducted. Potentiodynamic and galvanostatic tests in a 3.5% NaCl aqueous solution, XRD, SEM and energy-dispersive X-ray spectroscopy (EDX) analysis were performed. For annealed and specimens with a low RS magnitude, the dominant observation was pit initiation from existing pores and growth in the build direction. For specimens with medium RS level, SCC initiation from pore sites and propagation along melt pool boundaries and for specimens with the highest detected RS, crack initiation from melt pool boundaries, grains, machining marks, and pore sites were observed. Cracks propagated in different directions, i.e., along melt pool boundaries, near-surface transgranular, and transgranular through columnar microstructure. Galvanostatic tests showed three distinctive regions that corresponded to crack and pit initiation and growth. The synergistic effect of high dislocation density along melt pool boundaries, stress concentration in pore sites, molybdenum segregation, and surface RS was the cause of SCC susceptibility of specimens with high RS magnitude.

The effect of drug on a person may be different than expected because that drug interacts with another drug the person is taking (drug-drug interaction), food, beverages, dietary supplements the person is consuming (drug-nutrient/food interaction) or another disease the person has (drug-disease interaction). A drug interaction is a situation in which a substance affects the activity of a drug, i.e. the effects are increased or decreased, or they produce a new effect that neither produces on its own. These interactions may occur out of accidental misuse or due to lack of knowledge about the active ingredients involved in the relevant substances. Regarding food-drug interactions physicians and pharmacists recognize that some foods and drugs, when taken simultaneously, can alter the body's ability to utilize a particular food or drug, or cause serious side effects. Clinically significant drug interactions, which pose potential harm to the patient, may result from changes in pharmaceutical, pharmacokinetic, or pharmacodynamic properties. Some may be taken advantage of, to the benefit of patients, but more commonly drug interactions result in adverse drug events. Therefore it is advisable for patients to follow the physician and doctors instructions to obtain maximum benefits with least fooddrug interactions. The literature survey was conducted by extracting data from different review and original articles on general or specific drug interactions with food. This review gives information about various interactions between different foods and drugs and will help physicians and pharmacists prescribe drugs cautiously with only suitable food supplement to get maximum benefit for the patient.

Geographical Information System (GIS) is one of the recent innovation to ease users in locating places. GIS is a system designed to capture, store, manipulate, analyze, manage, and present all types of spatial or geographical data. It can also be used by customers in locating business premises. Research related to application of GIS in locating SME business premises especially in Malacca as a tourist state is still very limited. This study used a survey method to produce more generalizable findings. Quantitative procedures for data collection and analysis were rigorously applied to ensure reliability and validity of the findings. A total of 160 usable responses was collected from 400 distributed questionnaires. This study found all variables significant and therefore, all hypotheses were accepted. In the future, researchers may extend this study to other areas to understand more about current usage patterns and reasons for its adoption. Studies which cover non-Malaysian respondents and different education levels can further document different perspectives of GIS adoption.

Background: Pakistan is one of the two countries endemic for wild poliovirus type 1 (WPV1). Active clinical and environmental wastewater surveillance along with laboratory investigation is an integral and primary component of the polio eradication strategies. The current study is mainly focused on the virological data to understand the current epidemiology of WPV1 in Pakistan during 2019-2022., Methods: 141,037 stool specimens of patients reported with acute flaccid paralysis (AFP) and 3,171 wastewater samples were tested for poliovirus detection using cell culture and PCR. Phylogenetic analysis of WPV1 was performed using MEGA and Nextstrain., Results: Poliovirus isolates were classified into 15 distinct genetic clusters with multiple transmission lineages. Spatio-temporal trends indicated a significant decline in the incidence of poliomyelitis reported in 58 districts in 2019 to just 3 in 2022. The historical reservoirs in Peshawar, Quetta, and Karachi successfully eliminated the indigenous transmission chains of wild poliovirus active there for years., Conclusions: Our findings reinforce the evolving epidemiology of poliovirus in Pakistan which is now confined to South KP. All historically known reservoirs in Peshawar, Karachi and Quetta blocs are now free of poliovirus. Intensified clinical and environmental surveillance should be maintained to eliminate the very few remaining transmission lineages and certify the poliovirus eradication by 2026., (© The Author(s) 2024. Published by Oxford University Press on behalf of Infectious Diseases Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Widespread surveillance, rapid detection, and appropriate intervention will be critical for successful eradication of poliovirus. Using deployable next-generation sequencing (NGS) approaches, such as Oxford Nanopore Technologies' MinION, the time from sample to result can be significantly reduced compared to cell culture and Sanger sequencing. We developed piranha (poliovirus investigation resource automating nanopore haplotype analysis), a 'sequencing reads-to-report' solution to aid routine poliovirus testing of both stool and environmental samples and alleviate the bioinformatic bottleneck that often exists for laboratories adopting novel NGS approaches. Piranha can be used for efficient intratypic differentiation of poliovirus serotypes, for classification of Sabin-like polioviruses, and for detection of wild-type and vaccine-derived polioviruses. It produces interactive, distributable reports, as well as summary comma-separated values files and consensus poliovirus FASTA sequences. Piranha optionally provides phylogenetic analysis, with the ability to incorporate a local database, processing from raw sequencing reads to an interactive, annotated phylogeny in a single step. The reports describe each nanopore sequencing run with interpretable plots, enabling researchers to easily detect the presence of poliovirus in samples and quickly disseminate their results. Poliovirus eradication efforts are hindered by the lack of real-time detection and reporting, and piranha can be used to complement direct detection sequencing approaches., Competing Interests: None declared., (© The Author(s) 2024. Published by Oxford University Press.)

Direct detection by PCR of poliovirus RNA in stool samples provides a rapid diagnostic and surveillance tool that can replace virus isolation by cell culture in global polio surveillance. The sensitivity of direct detection methods is likely to depend on the choice of RNA extraction method and sample volume. We report a comparative analysis of 11 nucleic acid extraction methods (7 manual and 4 semiautomated) for poliovirus molecular detection using stool samples ( n  = 59) that had been previously identified as poliovirus positive by cell culture. To assess the effect of RNA recovery methods, extracted RNA using each of the 11 methods was tested with a poliovirus-specific reverse transcription-quantitative PCR (RT-qPCR), a pan-poliovirus RT-PCR (near-whole-genome amplification), a pan-enterovirus RT-PCR (entire capsid region), and a nested VP1 PCR that is the basis of a direct detection method based on nanopore sequencing. We also assessed extracted RNA integrity and quantity. The overall effect of extraction method on poliovirus PCR amplification assays tested in this study was found to be statistically significant ( P < 0.001), thus indicating that the choice of RNA extraction method is an important component that needs to be carefully considered for any diagnostic based on nucleic acid amplification. Performance of the methods was generally consistent across the different assays used. Of the 11 extraction methods tested, the MagMAX viral RNA isolation kit used manually or automatically was found to be the preferable method for poliovirus molecular direct detection considering performance, cost, and processing time. IMPORTANCE Poliovirus, the causative agent of poliomyelitis, is a target of global eradication led by the World Health Organization since 1988. Direct molecular detection and genomic sequencing without virus propagation in cell culture is arguably a critical tool in the final stages of polio eradication. Efficient recovery of good-quality viral RNA from stool samples is a prerequisite for direct detection by nucleic acid amplification. We tested 11 nucleic acid extraction methods to identify those facilitating sensitive, fast, simple, and cost-effective extraction, with flexibility for manual and automated protocols considered. Several different PCR assays were used to compare the recovered viral RNA to test suitability for poliovirus direct molecular detection. Our findings highlight the importance of choosing a suitable RNA extraction protocol and provide useful information to diagnostic laboratories and researchers facing the choice of RNA extraction method for direct molecular virus detection from stool.

Background: Elimination of poliovirus in Pakistan and Afghanistan is challenged by notions against the role of oral poliovirus vaccine (OPV) in eradicating contemporary wild poliovirus (WPV) strains., Methods: A total of 1055 WPV type 1 (WPV1) strains isolated between 2013 and 2018 were categorized into 68 antigenic groups and tested for neutralization by OPV-derived antibodies. Molecular docking was conducted to determine neutralization efficiency of antibodies against WPV. The clinical significance of WPV1 variants was assessed to ascertain their role in patient outcomes., Results: We found that 88% of WPV1 strains isolated from paralytic children belonged to a single antigenic lineage identical to the WPV1 strain detected in 1993. WPV1 antigenic variants were effectively neutralized by OPV-derived antibodies, with geometric mean titers comparable to the neutralization titers found for 3 strains in OPV (OPV1-3, 7.96-9.149 [95% confidence interval, 6.864-10.171]; WPV1 strains, 7.542-8.786 [6.493-9.869]). Docking examination underscored a strong antigen-antibody interaction despite variations within the viral protein 1 epitopes. There was no significant association (P = .78) with clinical prognosis among patients infected with antigenically diverse WPV1 strains and patient outcomes, including death., Conclusions: Our findings substantiate the robustness of OPV for neutralizing the contemporary WPV1 strains endemic in Pakistan and Afghanistan. Vaccination coverage must be augmented to achieve early eradication., Competing Interests: Potential conflicts of interest. All authors: No reported conflicts. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed., (© The Author(s) 2021. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Lower respiratory illness is one of the leading causes of death among children in low- and high-income countries. Human metapneumovirus (hMPV) is a key contributor to respiratory illnesses commonly reported among children and causes serious clinical complications ranging from mild respiratory infections to severe lower respiratory tract anomalies mainly in the form of bronchiolitis and pneumonia. However, due to the lack of a national surveillance system, the clinical significance of hMPV remains obscure in the Pakistani population. This study was conducted to screen throat swabs samples collected from 127 children reported with respiratory symptoms at a tertiary care hospital in Islamabad. Out of 127, 21 (16.5%) samples were positive for hMPV with its genotype distribution as A2a (10%), A2b (20%), B1 (10%), and B2 (60%). Phylogenetic analysis showed that the hMPV viruses were closely related to those reported from neighboring countries including India and China. This work will contribute to a better understanding of this virus, its diagnosis, and the handling of patients in clinical setups. Further studies at a large-scale are warranted for a better understanding of the disease burden and epidemiology of hMPV in Pakistan., (© 2022. The Author(s).)

Chikungunya virus (CHIKV) is considered a public health problem due to its rapid spread and high morbidity. In 2016-2017 an outbreak of CHIKV was occurred in Pakistan but the data regarding the genomic diversity of CHIKV was not reported. Hence, the current study aimed to determine the genetic diversity of CHIKVs in Pakistan. A cross sectional study was carried out using sera of infected CHIKV patients (n = 1549) during the outbreak in Pakistan (2016-2018). Nucleotide sequencing of non-structural genes of CHIKV from eight isolates were performed followed by phylogenetic analysis using Bayesian method. Phylogenetic analysis suggested that the Pakistani CHIKV strains belonged to Indian Ocean Lineage (IOL) of genotype ECSA and C1.3a clade. Furthermore, the Pakistani isolates showed several key mutations (nsP2-H130Y, nsP2-E145D, nsP4-S55N and nsP4- R85G) corresponding to mutations reported in 2016 Indian strains of CHIKV. The molecular analysis revealed high evolutionary potential of CHIKV strains as well as better understanding of enhanced virulence and pathogenesis of this outbreak. The study highlights the need to continue surveillance in order to understand viral diversity over time and to devise preventive measures to limit diseases transmission in the region., Competing Interests: The authors have declared that no competing interests exist.

The chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus, which has infected millions of people in Africa, Asia, Americas, and Europe since it remerged in India and Indian Ocean regions in 2005-2006. The purpose of this study was to evaluate the genetic diversity and evolutionary changes in CHIKV from 2016 to 2018 in Pakistan. Blood specimens were collected and processed following the Centers for Disease Control and Prevention Trioplex Protocol. Sequencing and phylogenetic analysis of complete coding sequence of representative isolates from the CHIKV outbreak was carried out during December 2016 to July 2018, a total of 1549 samples were received, out of which 50% (n = 774) were found positive for CHIKV RNA. Mean age of chikungunya positive patients was 31.8 ± 15.7 years and most affected were between 21 and 40 years of age. The Pakistan CHIKV strains clustered with the Indian Ocean sublineage of East/Central/South African with cocirculation of some variants In the structural proteins region, two noteworthy changes (A226V and D284E) were observed in the membrane fusion glycoprotein E1. Key substitutions in the neutralizing epitopes site and a few changes indicative of adaptive to other insect cells were also detected in Pakistani strains. This study provides the emerging trend of CHIKV in the country for early identification of potential variants of high virulence and preventive measures for vector borne disease especially in the endemic areas., (© 2021 Wiley Periodicals LLC.)

The ongoing COVID-19 pandemic is caused by SARs-CoV-2. The virus is transmitted from person to person through droplet infections i.e. when infected person is in close contact with another person. In January 2020, first report of detection of SARS-CoV-2 in faeces, has made it clear that human wastewater might contain this virus. This may illustrate the probability of environmentally facilitated transmission, mainly the sewage, however, environmental conditions that could facilitate faecal oral transmission is not yet clear. We used existing Pakistan polio environment surveillance network to investigate presence of SARs-CoV-2 using three commercially available kits and E-Gene detection published assay for surety and confirmatory of positivity. A Two-phase separation method is used for sample clarification and concentration. An additional high-speed centrifugation (14000Xg for 30 min) step was introduced, prior RNA extraction, to increase viral RNA yield resulting a decrease in Cq value. A total of 78 wastewater samples collected from 38 districts across Pakistan, 74 wastewater samples from existing polio environment surveillance sites, 3 from drains of COVID-19 infected areas and 1 from COVID 19 quarantine center drainage, were tested for presence of SARs-CoV-2. 21 wastewater samples (27%) from 13 districts turned to be positive on RT-qPCR. SARs-COV-2 RNA positive samples from areas with COVID 19 patients and quarantine center strengthen the findings and use of wastewater surveillance in future. Furthermore, sequence data of partial ORF 1a generated from COVID 19 patient quarantine center drainage sample also reinforce our findings that SARs-CoV-2 can be detected in wastewater. This study finding indicates that SARs-CoV-2 detection through wastewater surveillance has an epidemiologic potential that can be used as supplementary system to monitor viral tracking and circulation in cities with lower COVID-19 testing capacity or heavily populated areas where door-to-door tracing may not be possible. However, attention is needed on virus concentration and detection assay to increase the sensitivity. Development of highly sensitive assay will be an indicator for virus monitoring and to provide early warning signs., Competing Interests: The authors have declared that no competing interests exist.

Depleting supplies of fossil fuel, regular price hikes of gasoline and environmental deterioration have necessitated the search for economic and eco-benign alternatives of gasoline like lignocellulosic biomass. However, pre-treatment of such biomass results in development of some phenolic compounds which later hinder the depolymerisation of biomass by cellulases and seriously affect the cost effectiveness of the process. Dephenolification of biomass hydrolysate is well cited in literature. However, elimination of phenolic compounds from pretreated solid biomass is not well studied. The present study was aimed to optimize dephenoliphication of wheat straw using various alkalis i.e., Ca(OH) 2 and NH 3 ; acids i.e., H 2 O 2 , H 2 SO 4 , and H 3 PO 4 ; combinations of NH 3 + H 3 PO 4 and H 3 PO 4 + H 2 O 2 at pilot scale to increase enzymatic saccharification yield. Among all the pretreatment strategies used, maximum reduction in phenolic content was observed as 66 mg Gallic Acid Equivalent/gram Dry Weight (GAE/g DW), compared to control having 210 mg GAE/g DW using 5% (v/v) combination of NH 3 +H 3 PO 4 . Upon subsequent saccharification of dephenoliphied substrate, the hydrolysis yield was recorded as 46.88%. Optimized conditions such as using 1%+5% concentration of NH 3 + H 3 PO 4 , for 30 min at 110°C temperature reduced total phenolic content (TPC) to 48 mg GAE/g DW. This reduction in phenolic content helped cellulases to act more proficiently on the substrate and saccharification yield of 55.06% was obtained. The findings will result in less utilization of cellulases to get increased yield of saccharides by hydrolyzing wheat straw, thus, making the process economical. Furthermore, pilot scale investigations of current study will help in upgrading the novel process to industrial scale., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Haq, Nawaz, Liaqat, Arshad, Fan, Sun, Zhou, Xu, Akram and Jiang.)

Background: Fruit juice clarification is a challenging aspect of beverage industry which needs to be addressed for economical and hygienic production of fruit juices., Objective: Current study is focused on the complete purification, characterization and thermodynamic analysis of an efficient mannanase enzyme to analyze its applicability in biological clarification fruit juice., Methods: Mannanase production using Aspergillus awamori IIB037 in a 25 L stirred fermenter at pre optimized reaction conditions was carried out. Enzyme purification was carried out via series of steps. Characterization of enzyme along with kinetics and thermodynamic studies was conducted. Purified and characterized enzyme was assessed for its applicability in fruit juice clarification through clarification experiments on fresh apple juice., Results: Purification fold of 3.98 was obtained along with 86.80% purification yield of mannanase with specific activity of 158.16 U/mg. The molecular size of purified enzyme was determined as 66 kDa. The enzyme depicted 56% residual activity at 60°C after 8 hrs. Thermodynamic studies of an enzyme revealed enthalpy of activation (ΔH) and activation energy (Ea) as 30.53KJ/mol, 27.76KJ/mol, respectively. The enzyme activity increased in the presence of ß-mercaptoethanol surprisingly. On the other hand, methyl alcohol, ethanol, Hg2+ and Cu2+ inhibited enzyme activity. The enzyme showed Km and Vmax values of 11.07 mM and 19.08 μM min-1 for Locust Bean Gum (LBG) under optimal conditions. Juice treated with mannanase showed decrease in absorbance and increase in reducing sugar content., Conclusion: The current study demonstrated that mannanase from Aspergillus awamori in its purified form has significant characteristics to be employed industrially for juice clarification., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: Pakistan is among 3 countries endemic for wild poliovirus type 1 (WPV1) circulation that are still struggling for eradication of poliomyelitis. Active clinical and environmental surveillance with meticulous laboratory investigations provide insights into poliovirus transmission patterns and genomic diversity to inform decisions for strategic operations required to achieve eradication., Methods: We analyzed epidemiological and virological data to comprehend the current epidemiological status of WPV1 in Pakistan during 2015-2017. Stool specimens of patients with acute flaccid paralysis (AFP) and sewage samples collected from 60 environmental sites were tested. Viral culturing, intratypic differentiation by real-time polymerase chain reaction, and nucleic acid sequencing of the VP1 region of the poliovirus genome to determine genetic relatedness among WPV1 strains were applied., Results: Poliovirus isolates were grouped into 11 distinct clusters, which had ≥95% nucleotide homology in the VP1 coding region. Most of the poliovirus burden was shared by 3 major reservoirs: Karachi, Peshawar, and Quetta block (64.2% in 2015, 75.4% in 2016, and 76.7% in 2017)., Conclusions: Environmental surveillance reveals importations and pockets of unimmunized children that dictate intensive target mop-up campaigns to contain poliovirus transmission. A decrease in the number of orphan isolates reflects effective combination of AFP and environmental surveillance in Pakistan. The genetic data reflect sustained transmission within reservoir areas, further expanded by periodic importations to areas of high immunity reflected by immediate termination of imported viruses. Improved immunization coverage with high-quality surveillance is vital for global certification of polio eradication., (© The Author(s) 2019. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Global poliovirus surveillance involves virus isolation from stool and environmental samples, intratypic differential (ITD) by PCR, and sequencing of the VP1 region to distinguish vaccine (Sabin), vaccine-derived, and wild-type polioviruses and to ensure an appropriate response. This cell culture algorithm takes 2 to 3 weeks on average between sample receipt and sequencing. Direct detection of viral RNA using PCR allows faster detection but has traditionally faced challenges related to poor sensitivity and difficulties in sequencing common samples containing poliovirus and enterovirus mixtures. We present a nested PCR and nanopore sequencing protocol that allows rapid (<3 days) and sensitive direct detection and sequencing of polioviruses in stool and environmental samples. We developed barcoded primers and a real-time analysis platform that generate accurate VP1 consensus sequences from multiplexed samples. The sensitivity and specificity of our protocol compared with those of cell culture were 90.9% (95% confidence interval, 75.7% to 98.1%) and 99.2% (95.5% to 100.0%) for wild-type 1 poliovirus, 92.5% (79.6% to 98.4%) and 98.7% (95.4% to 99.8%) for vaccine and vaccine-derived serotype 2 poliovirus, and 88.3% (81.2% to 93.5%) and 93.2% (88.6% to 96.3%) for Sabin 1 and 3 poliovirus alone or in mixtures when tested on 155 stool samples in Pakistan. Variant analysis of sequencing reads also allowed the identification of polioviruses and enteroviruses in artificial mixtures and was able to distinguish complex mixtures of polioviruses in environmental samples. The median identity of consensus nanopore sequences with Sanger or Illumina sequences from the same samples was >99.9%. This novel method shows promise as a faster and safer alternative to cell culture for the detection and real-time sequencing of polioviruses in stool and environmental samples., (Copyright © 2020 Shaw et al.)

During December 2016-May 2017, an outbreak of chikungunya virus infection occurred across Pakistan. The East/Central/South African genotype was predominant. This study provides baseline data on the virus strain and emphasizes the need for active surveillance and implementation of preventive interventions to contain future outbreaks.

Despite the availability of an effective vaccine, the measles virus continues to cause significant morbidity and mortality in children worldwide. Molecular characterization of wild-type measles strains is an invaluable component of epidemiological studies or surveillance systems that provides important information pertinent to outbreak linkages and transmission pathways. Serum samples and throat swabs were collected from suspected measles cases from the Punjab province of Pakistan (2013-2015) and further tested for measles immunoglobulin M (IgM) through enzyme-linked immunosorbent assay and reverse-transcriptase polymerase chain reaction for molecular characterization. Among the total of 5415 blood samples, 59% tested positive for measles IgM. Males had a higher infection rate (55%) than females (45%), and the highest frequency of positive cases (63%) was found in the age group of 0 to 5 years. Partial sequencing of the nucleoprotein gene showed that 27 strains belonged to the B3 genotype, whereas 2 viruses were identified as D4. On phylogenetic analysis, Pakistani B3 strains were found to be closely related to previously reported indigenous strains and those from neighboring countries of Iran and Qatar. This is the first report on the detection of the measles B3 genotype from Punjab, Pakistan. The current study shows a high burden of measles infections in Punjab province owing to poor routine immunization coverage in major cities. It is imperative that national health authorities adopt strategic steps on an urgent basis for improvement of routine immunization coverage. Molecular epidemiology of the measles viruses circulating in different parts of the country can provide useful data to manage future outbreaks., (© 2018 Wiley Periodicals, Inc.)

Rotavirus A species (RVA) is the leading cause of severe diarrhea among children in both developed and developing countries. Among different RVA G types, humans are most commonly infected with G1, G2, G3, G4 and G9. During 2003-2004, G3 rotavirus termed as "new variant G3" emerged in Japan that later disseminated to multiple countries across the world. Although G3 rotaviruses are now commonly detected globally, they have been rarely reported from Pakistan. We investigated the genetic diversity of G3 strains responsible RVA gastroenteritis in children hospitalized in Rawalpindi, Pakistan during 2014. G3P[8] (18.3%; n = 24) was detected as the most common genotype causing majority of infections in children less than 06 months. Phylogenetic analysis of Pakistani G3 strains showed high amino acid similarity to "new variant G3" and G3 strains reported from China, Russia, USA, Japan, Belgium and Hungary during 2007-2012. Pakistani G3 strains belonged to lineage 3 within sub-lineage 3d, containing an extra N-linked glycosylation site compared to the G3 strain of RotaTeqTM. To our knowledge, this is the first report on the molecular epidemiology of G3 rotavirus strains from Pakistan and calls for immediate response measures to introduce RV vaccine in the routine immunization program of the country on priority.

An outbreak of dengue fever was reported in Malakand district, Khyber Pakhtunkhwa (KP) province of Pakistan during 2015. Detection of viral RNA by real-time PCR confirmed dengue virus serotype-3 (DENV-3) to be the causative agent causing the outbreak. Phylogenetic analysis based on partial E-NS1 gene sequences showed that the DENV-3 viruses belonged to genotype III with maximum homology with the dengue-3 strains previously reported from Pakistan and India. Our current report provides updated information on molecular epidemiology and phylogenetic analysis of dengue virus serotypes responsible for 2015 outbreak in KP., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Pneumonia remains a leading cause of morbidity and mortality in developing countries. Comprehensive surveillance data are needed to review the prevention and control strategies. We conducted active surveillance of acute lower respiratory infections among children aged <2 years hospitalized at two hospitals of Islamabad, Pakistan. Viral etiology was determined using real-time PCR on respiratory specimens collected during March 2011-April 2012. The overall mean age was 7.83 ± 5.25 months while no statistical difference between age or sex distribution of patients with positive and negative viral etiology (p > 0.05). The average weight of the study group was 6.1 ± 2.25 kg. ≥1 viral pathogens were detected in 75% cases. Major respiratory viruses included RSV-A: 44%, RSV-B: 23%, Influenza-A: 24.5%, Influenza-B: 7%, Adenovirus: 8.4% and HmPV: 5.2%. A single, dual or multiple viral pathogens were detected in 43%, 27% and 5.2% patients respectively. Common symptoms were cough (95%), apnoea (84%), fever (78%), wheeze (64.5%), nasal congestion (55%) and rhinorrhea (48%). Among the RSV positive cases, 2-6 months age group had highest detection rate for RSV-A (30%, n = 21/69) and RSV-B (20%, n = 14/69) while patients infected with Influenza-A were in 2.1-6 months age group (61%, 23/38). Statistically significant difference was observed between RSV-positive and negative cases for nutrition status (p = 0.001), cigarette/wood smoke exposure (p = 0.001) and concomitant clinical findings. Most patients had successful outcome on combination therapy with bronchodilators, inhaled steroids and antibiotics. Our findings underscore high burden of ALRI in Pakistan. Interventions targeting viral pathogens coupled with improved diagnostic approaches are critical for better prevention and control.

From 2013 to 2015, the National Institute of Health, Pakistan, received 1,270 blood samples of suspected dengue cases reported from inpatient and outpatient departments of various hospitals in Khyber Pakhtunkhwa (KPK) province. In this study, we determined the circulating dengue virus (DENV) serotypes using real-time reverse transcriptase (RT)-PCR to understand the serotype-based epidemiology of DENV. All four serotypes (DENV-1 [6%], DENV-2 [33%], DENV-3 [47%], and DENV-4 [0.1%]) were found circulating during the study period. Our findings suggest the need for an active surveillance system coupled with the laboratory diagnosis, especially in the chronic endemic areas of the country. Public awareness programs are needed for effective control and prevention of outbreaks in the future., Competing Interests: Authors' Disclosures of Potential Conflicts of Interest: No potential conflicts of interest relevant to this article were reported.