Consensus statements for influenza awareness, prevention, and vaccination in Pakistan.

INTRODUCTION: Influenza is a serious underestimated viral infection in Pakistan and influenza vaccination and vaccination awareness are low. The current work aimed to develop consensus on influenza epidemiology, prevention, vaccination, and awareness in Pakistan. METHODOLOGY: A systematic literature search was conducted to develop recommendations on influenza vaccines in Pakistan. Experts' feedback was incorporated using the modified Delphi method. A three-step process was used, with 18 experts from different specialties from Pakistan who participated in voting rounds to achieve a minimum 75% agreement level. RESULTS: Pakistan has a low-immunization-rate and is susceptible to serious influenza outbreaks and influenza-related complications. Influenza circulates year-round in Pakistan but peaks during January and February. The subtype A/H1N1 is predominant. The experts urged vaccination in all individuals ≥ 6 months of age and with no contraindications. They highlighted special considerations for those with comorbidities and specific conditions. The experts agreed that the inactivated influenza vaccine is safe and efficient in pregnant women, immunocompromised, and comorbid respiratory and cardiovascular patients. Finally, the experts recommended conducting promotional and educational programs to raise awareness on influenza and vaccination. CONCLUSIONS: This is the first regional consensus on influenza and influenza vaccination in Pakistan with experts' recommendations to increase influenza vaccination and decrease influenza cases and its associated detrimental effects.

Enhancing curve and surface applications with trigonometric polynomial basis functions.

This article primarily focuses on the utilization and importance of parametric curves in surface design. It delves into the construction and applications of parametric curves, exploring the implementation of trigonometric polynomial basis functions that possess two shape parameters. Initially, these basis functions are employed in constructing both rational and non-rational curves. Later, they are employed to define the surfaces generated by these curves. The discussion includes rational surfaces, tensor product surfaces, and various specialized surfaces. The aim is to provide a comprehensive understanding of the role and potential of parametric curves in surface design.

Chitinase of Trichoderma longibrachiatum for control of Aphis gossypii in cotton plants.

Chitinase-producing fungi have now engrossed attention as one of the potential agents for the control of insect pests. Entomopathogenic fungi are used in different regions of the world to control economically important insects. However, the role of fungal chitinases are not well studied in their infection mechanism to insects. In this study, Chitinase of entomopathogenic fungi Trichoderma longibrachiatum was evaluated to control Aphis gossypii. For this purpose, fungal chitinase (Chit1) gene from the genomic DNA of T. longibrachiatum were isolated, amplified and characterised. Genomic analysis of the amplified Chit1 showed that this gene has homology to family 18 of glycosyl hydrolyses. Further, Chit1 was expressed in the cotton plant for transient expression through the Geminivirus-mediated gene silencing vector derived from Cotton Leaf Crumple Virus (CLCrV). Transformed cotton plants showed greater chitinase activity than control, and they were resistant against nymphs and adults of A. gossypii. About 38.75% and 21.67% mortality of both nymphs and adults, respectively, were observed by using Chit1 of T. longibrachiatum. It is concluded that T. longibrachiatum showed promising results in controlling aphids by producing fungal chitinase in cotton plants and could be used as an effective method in the future.

Quality of Life Index in Patients with Vitiligo.

OBJECTIVE: To ascertain the quality of life (QoL) impairment among the Pakistani population with vitiligo and to determine the relationship between sociodemographic and clinical characteristics. STUDY DESIGN: Cross-sectional study. Place and Duration of the Study: Dermatology Outpatients at the Aga Khan University in collaboration with outpatients of seven tertiary care hospitals of Sindh, Punjab, KPK, Balochistan, and AJK to collect data from March 2015 to April 2019. METHODOLOGY: All clinically diagnosed patients of vitiligo, who signed consent and assent forms, were included in the study. A validated 25-item, QoL scale for vitiligo was used. Socio-economic status of the patients, clinical assessment of the disease and patients' engagement in social and domestic lives was noted. RESULTS: Five hundred and seventy-three patients were enrolled in the study, having mean age 29.8 ± 16.2 years. In 306 (53.4%) males and 267 (46.65%) females; 21.8% were below 18 years. Mean vitiligo QoL index was 38.4 ± 11.8. Patients of vitiligo with disease duration 5-10 years, those affected on exposed parts, more than five body sites, rapidly progressing disease and of female gender had a higher impairment of quality of life. These scores were found significantly higher as compared to other levels of these parameters (p<0.05). CONCLUSION: Patients with vitiligo experience low self-esteem. The disease adversely affects their quality of life. The authors recommend the use of disease-specific instruments to assess the quality of life which enables the treating physician to devise best possible management plan individually. KEY WORDS: Vitiligo, Quality of life, Pakistan, Vitiligo life quality index.

Bio-fabrication of Zinc Oxide nanoparticles to rescue Mung Bean against Cercospora leaf spot disease.

Plant disease management using nanotechnology is evolving continuously across the world. The purpose of this study was to determine the effect of different concentrations of green synthesized zinc oxide nanoparticles (ZnO NPs) using Trachyspermum ammi seed extract on Cercospora leaf spot disease in mung bean plants under in-vitro and in-planta conditions. Additionally, the effects on mung bean agronomic and physiological parameters were also assessed. The green synthesized ZnO NPs were characterized using UV-visible spectroscopy, Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and Scanning electron microscopy (SEM). Green synthesized NPs were tested for their ability to inhibit fungal growth at five different concentrations under in-vitro experiment. After 7 days of inoculation, ZnO NPs (1200 ppm) inhibited mycelial growth substantially (89.86% ± 0.70). The in-planta experiment showed statistically significant result of disease control (30% ± 11.54) in response to 1200 ppm ZnO NPs. The same treatment showed statistically significant improvements in shoot length, root length, number of leaves, number of pods, shoot fresh weight (28.62%), shoot dry weight (85.18%), root fresh weight (38.88%), and root dry weight (38.88%) compared to the control. Our findings show that green synthesized ZnO NPs can control Cercospora canescens in mung bean, pointing to their use in plant disease control and growth enhancement.

Progressive Thoracic Kyphoscoliosis leading to Paraplegia in a Child with Neurofibromatosis Type-1.

In this case report we describe a case of a 13-year male with neurofibromatosis type-1, who had been suffering from progressive thoracic kyphoscoliosis that resulted in spinal cord compression and paraplegia. The report is meant to highlight the importance of timely detection and adequate follow-up of spinal deformities, associated with neurofibromatosis. Such deformities, if not effectively addressed, may progress corresponding to skeletal growth and lead to secondary problems, as was seen in our patient, who ultimately developed paraplegia. Key Words: Neurofibromatosis, Kyphoscoliosis, Paraplegia.

Recent advances in combinatorial cancer therapy via multifunctionalized gold nanoparticles.

The diverse behavior of nanogold in the therapeutic field is related to its unique size and shape. Nanogold offers improvements in modern diagnostic and therapeutic implications, increases disease specificity and targeted drug delivery, and is relatively economical compared with other chemotherapeutic protocols. The diagnosis of cancer and photothermal therapy improve drastically with the implementation of nanotechnology. Different types of nanoparticles, that is, gold silica nanoshells, nanorods and nanospheres of diverse shapes and geometries, are used widely in the photothermal therapy of cancerous cells and nodules. Numerous reviews have been published on the therapeutic applications of gold nanoparticles, but studies on combinatorial applications of nanogold in cancer therapy are limited. This review focuses on the combinatorial cancer therapy using optical properties of nanogold with different shapes and geometries, and their therapeutic applications in cancer diagnosis, photothermal therapy, cancer imaging and targeted drug delivery.

Comparison of Intralesional Meglumine Antimonite along with oral Itraconazole to Intralesional Meglumine Antimonite in the treatment of Cutaneous Leishmaniasis.

BACKGROUND & OBJECTIVE: Cutaneous leishmaniasis (CL) is endemic in developing countries like Pakistan. Pentavalent antimonials are still drug of choice, despite being toxic and intolerable for patients. Second line treatments have been extensively studied but the results of their efficacy are conflicting. This, to our knowledge, will be the first study in this regard. Our objective was to determine if combination of oral itraconazole with intralesional (IL) meglumine antimoniate (MA) reduces the duration of treatment for cutaneous leishmaniasis, as compared to intralesional MA alone. METHODS: A randomized controlled trial (single blinded) was carried out from August 2017 till December 2017 on 69 patients who fulfilled inclusion criteria. They were assigned to Group-A or B by lottery method. Group-A patients received IL MA once a week while Group-B received oral itraconazole 200mg, once daily, for six weeks along with similar regimen of IL MA as Group-A. The patients were assessed every three weeks by the blinded assessor till clinical cure was achieved. A follow up visit, two months after clinical cure was done to look for relapse of the disease. RESULTS: Thirty patients in Group-A and 35 patients in Group-B completed the study. At 3, 6, 9 and 12 weeks the patients were assessed for: no, partial or complete response and results of the two groups were compared for statistical significance. The p-values of 0.20, 0.57 and 0.11 at 3, 6 and 9 weeks, respectively, depict that there was no significant difference at any step of assessment between the two groups in terms of healing. The p values of each t test was>0.05 refuting the hypothesis. CONCLUSION: Combination of oral itraconazole with intralesional MA offered no benefit over intralesional MA alone in the management of cutaneous leishmaniasis in terms of duration of therapy.

Safety and efficacy of miltefosine in cutaneous leishmaniasis: An open label, non-comparative study from Balochistan.

BACKGROUND & OBJECTIVE: Cutaneous Leishmaniasis (CL) is endemic in Baluchistan and treated traditionally with Meglumine antimoniate. Miltefosine appears appealing therapy in cutaneous Leishmaniasis. Our objective was to evaluate safety and efficacy of Miltifossine in treatment of cutaneous Leishmaniasis. METHODS: This experimental study was conducted from 10 September 2017 to 10 May 2018 at Combined Military Hospital Quetta. Total of 42 patients were recruited by purposive sampling technique. Lesional skin smears were stained with giemsa for Leishmania amastigotes under magnification (100 x).Complete blood count, serum urea, creatinine, bilirubin, aspartate aminotransferases (AST), alanine aminotransferase (ALT) were done at the beginning of treatment and then weekly, thereafter. Cap Miltefosine 50 mg (2.5mg/kg) were given as directly observed therapy .Daily observation during treatment phase was done for clinical side effects of therapy. Clinical response was documented at two weeks then at eight weeks. Photographs were taken before and after the therapy. Data was analyzed by SPSS 16. RESULTS: Complete clinical response was observed in 39 (92.9%) patients and partial clinical response in 1(2.4%) patient. Two patients were lost to follow up at eight weeks. No significant derangements in laboratory profile were noted before and after treatment. Mean duration of treatment was 23.47+SD 4.44 days. Sixteen patients (38.1%) took Miltefosine for 28 days, 12 (28.6%) for 21 days and 9 (25%) for 25 days. CONCLUSION: Miltefosine is safe and cost effective treatment for cutaneous Leishmaniasis. It is effective in CL cases not susceptible to antimony compounds.

Influenza vaccination situation in Middle-East and North Africa countries: Report of the 7th MENA Influenza Stakeholders Network (MENA-ISN).

BACKGROUND: The Middle East and North Africa (MENA) region faces a dual challenge with regard to influenza infection due to severe zoonotic influenza outbreaks episodes and the circulation of Northern Hemisphere human influenza viruses among pilgrims. METHODS: The MENA Influenza Stakeholder Network (MENA-ISN) was set-up with the aim of increasing seasonal influenza vaccination coverage by (i) enhancing evidence-based exchanges, and (ii) increasing awareness on the safety and benefits of seasonal vaccination. During the 7th MENA-ISN meeting, representatives from 8 countries presented their influenza surveillance, vaccination coverage and actions achieved and provided a list of country objectives for the upcoming 3 years. RESULTS: MENA-ISN countries share the goal to reduce influenza related morbidity and mortality. Participants admitted that lack of knowledge about influenza, its consequences in terms of morbidity, mortality and economy are the major barrier to attaining higher influenza vaccination coverage in their countries. The cost of the vaccine is another key barrier that could contribute to low vaccination coverage. Participants drew a list of strategic interventions to bridge gaps in the knowledge of influenza burden in this region. CONCLUSIONS: Participating countries concluded that despite an increase in vaccine uptake observed during the last few years, influenza vaccination coverage remains relatively low. Priority areas should be identified and action plans tailored to each country situation set-up to investigate the best way to move forward.

Molecular detection and characterization of respiratory syncytial virus B genotypes circulating in Pakistani children.

Respiratory syncytial virus (RSV) is the major cause of acute lower respiratory tract infections in young children, but very little is known about its epidemiology and circulating genotypes in Pakistan. This study analyzed the epidemiological and molecular characteristics of RSV B genotypes in Pakistani children below 5years with acute respiratory tract infections (ARIs) during three consecutive winter seasons from 2010 to 2013. A total of 1941 samples were analyzed for RSV infection by real time PCR and 24% (472/1941) samples were found positive out of which 22.3% (105/472) were sub-typed as RSV-B. The frequency of outpatient cases was higher (62.5%; 295/472) as compared to hospitalized patients (37.5%; 177/472). Patient ages ranged from 2month to 5years with a mean age of 1.48±1.2 (years) and a median age of 1year. Children below one year made up the highest percentage of enrolled subjects and male to female ratio of RSVB positive cases was nearly equivalent (1:1.1). The most common clinical symptoms were cough (96%), fever (80%) and sore throat (50%). All Pak RSVB strains ascribed to the BA genotype showing 91.9-97.1% and 86.2-95.3% homology at the nucleotide and amino acid levels respectively in comparison to BA prototype strain. On phylogenetic analysis, three genotypes of Pakistan RSV B viruses were observed; BA-9 and BA-10 which have been reported previously from other regions, and a third novel genotype assigned as BA-13 which formed a distinct cluster with protein length of 319 AA and showed 9-11 unique AA substitutions. All the RSV B isolates had two potential N-glycosylation sites in HVR2 of G protein and with heavy O-glycosylation of serine and threonine residues (G scores of 0.5-0.7). This study highlights the diversity of RSVB viruses and the significance of RSV as a dominant viral etiologic agent of pediatric ARI. It also emphasizes the need for continued molecular surveillance for early detection of prevalent and newly emerging genotypes to understand epidemiology of RSV infections in various regions of Pakistan.

Respiratory syncytial virus and influenza are the key viral pathogens in children <2 years hospitalized with bronchiolitis and pneumonia in Islamabad Pakistan.

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.

Strengthening the influenza vaccine virus selection and development process: Report of the 3rd WHO Informal Consultation for Improving Influenza Vaccine Virus Selection held at WHO headquarters, Geneva, Switzerland, 1-3 April 2014.

Despite long-recognized challenges and constraints associated with their updating and manufacture, influenza vaccines remain at the heart of public health preparedness and response efforts against both seasonal and potentially pandemic influenza viruses. Globally coordinated virological and epidemiological surveillance is the foundation of the influenza vaccine virus selection and development process. Although national influenza surveillance and reporting capabilities are being strengthened and expanded, sustaining and building upon recent gains has become a major challenge. Strengthening the vaccine virus selection process additionally requires the continuation of initiatives to improve the timeliness and representativeness of influenza viruses shared by countries for detailed analysis by the WHO Global Influenza Surveillance and Response System (GISRS). Efforts are also continuing at the national, regional, and global levels to better understand the dynamics of influenza transmission in both temperate and tropical regions. Improved understanding of the degree of influenza seasonality in tropical countries of the world should allow for the strengthening of national vaccination policies and use of the most appropriate available vaccines. There remain a number of limitations and difficulties associated with the use of HAI assays for the antigenic characterization and selection of influenza vaccine viruses by WHOCCs. Current approaches to improving the situation include the more-optimal use of HAI and other assays; improved understanding of the data produced by neutralization assays; and increased standardization of serological testing methods. A number of new technologies and associated tools have the potential to revolutionize influenza surveillance and response activities. These include the increasingly routine use of whole genome next-generation sequencing and other high-throughput approaches. Such approaches could not only become key elements in outbreak investigations but could drive a new surveillance paradigm. However, despite the advances made, significant challenges will need to be addressed before next-generation technologies become routine, particularly in low-resource settings. Emerging approaches and techniques such as synthetic genomics, systems genetics, systems biology and mathematical modelling are capable of generating potentially huge volumes of highly complex and diverse datasets. Harnessing the currently theoretical benefits of such bioinformatics ("big data") concepts for the influenza vaccine virus selection and development process will depend upon further advances in data generation, integration, analysis and dissemination. Over the last decade, growing awareness of influenza as an important global public health issue has been coupled to ever-increasing demands from the global community for more-equitable access to effective and affordable influenza vaccines. The current influenza vaccine landscape continues to be dominated by egg-based inactivated and live attenuated vaccines, with a small number of cell-based and recombinant vaccines. Successfully completing each step in the annual influenza vaccine manufacturing cycle will continue to rely upon timely and regular communication between the WHO GISRS, manufacturers and regulatory authorities. While the pipeline of influenza vaccines appears to be moving towards a variety of niche products in the near term, it is apparent that the ultimate aim remains the development of effective "universal" influenza vaccines that offer longer-lasting immunity against a broad range of influenza A subtypes.

Efficacy of clarithromycin in pityriasis rosea.

OBJECTIVE: To determine the efficacy of clarithromycin in the treatment of Pityriasis Rosea (PR). STUDY DESIGN: Double blind randomized controlled trial. PLACE AND DURATION OF STUDY: Dermatology OPD, Military Hospital, Rawalpindi, from July 2008 to July 2009. METHODOLOGY: Patients aged above 10 years, diagnosed with PR, were randomly assigned to two groups of 30 each to receive either clarithromycin or similar-looking placebo for one week. Neither the patient nor the treating physician knew to which group the patient belonged. Patients were assessed at 1, 2, 4 and 6 weeks after presentation and compared for complete, partial or no response. RESULTS: Among the 60 patients, no significant difference was found between the two groups at 2 weeks after presentation (p = 0.598). In the placebo group, complete response was seen in 20 (66.7%), partial response in 3 (10.0%) while no response was seen in 7 (23.3%). In clarithromycin group, there was complete response in 23 (76.7%), partial response in 3 (10.0%) and no response in 4 (13.3%) patients. CONCLUSION: Clarithromycin is not effective in treatment of pityriasis rosea.

Molecular characterization of circulating respiratory syncytial virus (RSV) genotypes in Gilgit Baltistan Province of Pakistan during 2011-2012 winter season.

Respiratory syncytial virus (RSV) is the major cause of acute lower respiratory tract infections in young children, but very little is known about its epidemiology and circulating genotypes in Pakistan. This study analyzed the epidemiological and molecular characteristics of RSV genotypes detected in Pakistani children less than 2 years of age with acute respiratory tract infections (ARIs) in a tertiary care hospital in Gilgit Baltistan (GB) province during 2011-12 winter season. RSV was detected in 75 out of 105 children presenting with acute respiratory infection. Male infants between 2-6 months age made up the highest percentage of RSV positive cases. Epidemiological factors such as pre-maturity, mean weight, clinical features and diagnosis when compared between RSV positive and negative groups were found to be statistically insignificant. Phylogenetic analysis classified all 75 of the RSV strains into 71 strains of subgroups A and 4 strains of subgroup B, respectively. Strains belonging to subgroups A and B were further subdivided into NA1/GA2 and BA, respectively. The nucleotide and deduced amino acid sequence identities were relatively high among these strains (>90%). Both RSV-A and RSV-B isolates had two potential N-glycosylation sites in HVR2 of G protein and with heavy O-glycosylation of serine and threonine residues (G scores of 0.5-0.7). This report highlights the significance of RSV as a dominant viral etiologic agent of pediatric ARIs, and need for continued molecular epidemiological surveys for early detection of prevalent strains and newly emerging genotypes to understand epidemiology of RSV infections in various regions of Pakistan.

Diagnostic and therapeutic implications of ST-segment elevation in lead aVR of 12 lead ECG during chest pain.

'aVR' is usually not the preferred lead to diagnose myocardial infarction in clinical settings, it is rather a neglected lead in this context. We describe the case of a 44 year old male who presented with short duration chest pain and ST segment elevation in lead 'aVR'. His left heart catheterization showed left main stem equivalent disease and totally occluded right coronary artery. Patient underwent emergency coronary artery bypass-grafting with favorable outcome. This case highlights the significance of ST segment elevation in lead aVR during chest pain both in diagnosis and management of patients with acute coronary syndrome.

Rheumatic mitral stenosis presenting as acute abdominal aortic occlusion and foot drop.

Systemic thromboembolism in not an infrequent complication of rheumatic mitral stenosis. We report a case of acute abdominal aortic occlusion due to rheumatic mitral stenosis, in the absence of atrial fibrillation, mimicking spinal cord compression. Systemic streptokinase given 48 hours after symptom's onset lead to rapid and dramatic recovery with limb salvage.