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Hypomagnesemia with secondary hypocalcemia is a rare autosomal-recessive disorder characterized by intense hypomagnesemia associated with hypocalcemia (HSH). Mutations in the TRPM6 gene, encoding the epithelial Mg 2+ channel TRPM6, have been proven to be the molecular cause of this disease. This study identified causal mutations in a 2-month-old male patient of hypomagnesemia from a consanguineous marriage. Biochemical analyses indicated the diagnosis of HSH due to primary gastrointestinal loss of magnesium. Whole exome sequencing of the trio (i.e. proband and both parents) was carried out with mean coverage of > 150×. ANNOVAR was used to annotate functional consequences of genetic variation from exome sequencing data. After variant filtering and annotation, a number of single nucleotide variants (SNVs) and 2 bp deletion at exon26:c.4402_4403delCT in TRPM6 gene were identified. This deletion which resulted in a novel frameshift mutation in exon 26 of this gene was confirmed by Sanger sequencing. With these investigations in hand, the patient was managed with magnesium sulphate. The patient remained asymptomatic and was developmentally and neurologically normal till his last follow up.

Electrocardiography (ECG) sensors play a vital role in the Internet of Medical Things, and these sensors help in monitoring the electrical activity of the heart. ECG signal analysis can improve human life in many ways, from diagnosing diseases among cardiac patients to managing the lifestyles of diabetic patients. Abnormalities in heart activities lead to different cardiac diseases and arrhythmia. However, some cardiac diseases, such as myocardial infarction (MI) and atrial fibrillation (Af), require special attention due to their direct impact on human life. The classification of flattened T wave cases of MI in ECG signals and how much of these cases are similar to ST-T changes in MI remain an open issue for researchers. This article presents a novel contribution to classify MI and Af. To this end, we propose a new approach called deep deterministic learning (DDL), which works by combining predefined heart activities with fused datasets. In this research, we used two datasets. The first dataset, Massachusetts Institute of Technology-Beth Israel Hospital, is publicly available, and we exclusively obtained the second dataset from the University of Malaya Medical Center, Kuala Lumpur Malaysia. We first initiated predefined activities on each individual dataset to recognize patterns between the ST-T change and flattened T wave cases and then used the data fusion approach to merge both datasets in a manner that delivers the most accurate pattern recognition results. The proposed DDL approach is a systematic stage-wise methodology that relies on accurate detection of R peaks in ECG signals, time domain features of ECG signals, and fine tune-up of artificial neural networks. The empirical evaluation shows high accuracy (i.e., ≤99.97%) in pattern matching ST-T changes and flattened T waves using the proposed DDL approach. The proposed pattern recognition approach is a significant contribution to the diagnosis of special cases of MI.

Background: Enteroviruses are common human pathogens occasionally associated with severe disease, notoriously paralytic poliomyelitis caused by poliovirus. Other enterovirus serotypes such as enterovirus A71 and D68 have been linked to severe neurological syndromes. New enterovirus serotypes continue to emerge, some believed to be derived from nonhuman primates. However, little is known about the circulation patterns of many enterovirus serotypes and, in particular, the detailed enterovirus composition of sewage samples., Methods: We used a next-generation sequencing approach analyzing reverse transcriptase polymerase chain reaction products synthesized directly from sewage concentrates., Results: We determined whole-capsid genome sequences of multiple enterovirus strains from all 4 A to D species present in environmental samples from the United Kingdom, Senegal, and Pakistan., Conclusions: Our results indicate complex enterovirus circulation patterns in human populations with differences in serotype composition between samples and evidence of sustained and widespread circulation of many enterovirus serotypes. Our analyses revealed known and divergent enterovirus strains, some of public health relevance and genetically linked to clinical isolates. Enteroviruses identified in sewage included vaccine-derived poliovirus and enterovirus D-68 stains, new enterovirus A71 and coxsackievirus A16 genogroups indigenous to Pakistan, and many strains from rarely reported serotypes. We show how this approach can be used for the early detection of emerging pathogens and to improve our understanding of enterovirus circulation in humans.

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.)

Poliovirus (PV) environmental surveillance (ES) plays an important role in the global eradication program and is crucial for monitoring silent PV circulation especially as clinical cases decrease. This study compared ES results using the novel bag-mediated filtration system (BMFS) with the current two-phase separation method. From February to November 2016, BMFS and two-phase samples were collected concurrently from twelve sites in Pakistan (n = 117). Detection was higher in BMFS than two-phase samples for each Sabin-like (SL) PV serotype (p<0.001) and wild PV type 1 (WPV1) (p = 0.065). Seventeen sampling events were positive for WPV1, with eight discordant in favor of BMFS and two in favor of two-phase. A vaccine-derived PV type 2 was detected in one BMFS sample but not the matched two-phase. After the removal of SL PV type 2 (SL2) from the oral polio vaccine in April 2016, BMFS samples detected SL2 more frequently than two-phase (p = 0.016), with the last detection by either method occurring June 12, 2016. More frequent PV detection in BMFS compared to two-phase samples is likely due to the greater effective volume assayed (1620 mL vs. 150 mL). This study demonstrated that the BMFS achieves enhanced ES for all PV serotypes in an endemic country., Competing Interests: The authors have declared that no competing interests exist.

Objectives: Automatic text classification techniques are useful for classifying plaintext medical documents. This study aims to automatically predict the cause of death from free text forensic autopsy reports by comparing various schemes for feature extraction, term weighing or feature value representation, text classification, and feature reduction., Methods: For experiments, the autopsy reports belonging to eight different causes of death were collected, preprocessed and converted into 43 master feature vectors using various schemes for feature extraction, representation, and reduction. The six different text classification techniques were applied on these 43 master feature vectors to construct a classification model that can predict the cause of death. Finally, classification model performance was evaluated using four performance measures i.e. overall accuracy, macro precision, macro-F-measure, and macro recall., Results: From experiments, it was found that that unigram features obtained the highest performance compared to bigram, trigram, and hybrid-gram features. Furthermore, in feature representation schemes, term frequency, and term frequency with inverse document frequency obtained similar and better results when compared with binary frequency, and normalized term frequency with inverse document frequency. Furthermore, the chi-square feature reduction approach outperformed Pearson correlation, and information gain approaches. Finally, in text classification algorithms, support vector machine classifier outperforms random forest, Naive Bayes, k-nearest neighbor, decision tree, and ensemble-voted classifier., Conclusion: Our results and comparisons hold practical importance and serve as references for future works. Moreover, the comparison outputs will act as state-of-art techniques to compare future proposals with existing automated text classification techniques., (Copyright © 2017 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.)

Text categorization has been used extensively in recent years to classify plain-text clinical reports. This study employs text categorization techniques for the classification of open narrative forensic autopsy reports. One of the key steps in text classification is document representation. In document representation, a clinical report is transformed into a format that is suitable for classification. The traditional document representation technique for text categorization is the bag-of-words (BoW) technique. In this study, the traditional BoW technique is ineffective in classifying forensic autopsy reports because it merely extracts frequent but discriminative features from clinical reports. Moreover, this technique fails to capture word inversion, as well as word-level synonymy and polysemy, when classifying autopsy reports. Hence, the BoW technique suffers from low accuracy and low robustness unless it is improved with contextual and application-specific information. To overcome the aforementioned limitations of the BoW technique, this research aims to develop an effective conceptual graph-based document representation (CGDR) technique to classify 1500 forensic autopsy reports from four (4) manners of death (MoD) and sixteen (16) causes of death (CoD). Term-based and Systematized Nomenclature of Medicine-Clinical Terms (SNOMED CT) based conceptual features were extracted and represented through graphs. These features were then used to train a two-level text classifier. The first level classifier was responsible for predicting MoD. In addition, the second level classifier was responsible for predicting CoD using the proposed conceptual graph-based document representation technique. To demonstrate the significance of the proposed technique, its results were compared with those of six (6) state-of-the-art document representation techniques. Lastly, this study compared the effects of one-level classification and two-level classification on the experimental results. The experimental results indicated that the CGDR technique achieved 12% to 15% improvement in accuracy compared with fully automated document representation baseline techniques. Moreover, two-level classification obtained better results compared with one-level classification. The promising results of the proposed conceptual graph-based document representation technique suggest that pathologists can adopt the proposed system as their basis for second opinion, thereby supporting them in effectively determining CoD., (Copyright © 2018 Elsevier Inc. All rights reserved.)

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.

The use of algal-bacterial symbiotic association establishes a sustainable and cost-effective strategy in wastewater treatment. Using municipal wastewater, the removal performances of inorganic nutrients (nitrogen and phosphorus) and organic pollutants were investigated by the co-culture system having different inoculum ratios (R) of suspended activated sludge to alginate-immobilized microalgae Chlorella vulgaris. The co-culture reactors with lower R ratios obtained more removal of nitrogen than in pure culture of C. vulgaris. The reactor with R = 0.5 (sludge/microalgae) showed the highest performance representing 66% removal after 24 h and 95% removal after 84 h. Phosphorus was completely eliminated (100%) in the co-culture system with inoculum ratios of 0.5 and 1.0 after 24 h and in the pure C. vulgaris culture after 36 h. The COD level was greatly reduced in the activated sludge reactor, while, it was increasing in pure C. vulgaris culture after 24 h of incubation. However, COD was almost stabilized after 24 h in the reactors with high R ratios such as 2.0, 5.0, and 10 due to the higher concentration of activated sludge. The growth of C. vulgaris was promoted from 0.03 g/L/d to 0.05 g/L/d in the co-culture of low inoculum ratios such as R = 0.5, implying that there exist an optimum inoculum ratio in the co-culture system in order to achieve efficient removal of nutrients., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

As a part of strategy to control diarrheal diseases, World Health Organization (WHO) recommends to include rotavirus vaccines in national immunization programs. Sentinel surveillance networks have been established to monitor rotavirus disease burden and genotype distribution in both pre and post vaccine era in many countries. Unfortunately, due to lack of proper surveillance programs, data on rotavirus disease burden and genotype distribution from Pakistan is scarce. We investigated 502 stool samples from children (<5years) hospitalized due to gastroenteritis in Rawalpindi, Pakistan during 2014 for the presence of group A rotavirus (RVA) and its genotypic diversity. Among 147 ELISA positive samples, 131 were successfully genotyped for RVA. Common G types detected were G1 (23.6%), followed by G3 (22.9%), G12 (19.8%), G2 (19.08%) and G9 (9.9%). The most common P-type was P[8] (41.2%), followed by P[6] (29%) and P[4] (28.24%). G3P[8] (17.55%) was the most prevalent genotype combination followed by G12P[6] (16.7%), G2P[4] (15.2%) and G1P[8] (14.5%). Mixed infection of rotavirus G-P types was also observed in 6% of samples. Phylogenetic analysis of VP7 and VP4 genes of Pakistani strains showed that G1, G2, G9 and P[4], P[6], P[8] were closely related to strains circulating worldwide as well as previously reported strains from Pakistan. Pakistani G12P[8] strains NIH-BBH-3981 and NIH-BBH-4003 belonged to lineage 3 cluster 3a along with strains from USA and Italy whereas G12P[6] strains NIH-BBH-3978, NIH-BBH-4052 and NIH-BBH-4444 were closely related to strains from Italy, Thailand, United Kingdom and with previously reported G12 strains from Pakistan within lineage 3 cluster 3b. This pre-vaccination data supports the need for RVA vaccine inclusion at our national level and will be helpful in assessing the effect of vaccination on RVA genotype diversity due to vaccine selection pressure once post-vaccination data becomes available., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The determination of heavy metals in blood is an important occupational environmental toxicology screening procedure. The aim of study was to determine the concentrations of Pb, Cd, Cr and Ni in blood samples of iron and steel foundry workplace exposed workers under routine clinical laboratory conditions. The method was employed for the quantitative determination of lead, cadmium, chromium and nickel in workplace environment particulate matter blood samples from iron and steel foundry workers and in unexposed controls. The results indicate that lead, chromium and nickel levels of the exposed workers are significantly higher those of the controls. Nickel Concentration (μg/L) in high P value=0.0306 and Chromium Concentration (μg/L) in P value=0.0295in worker population as compared to controls. Lead showed highest Absorption concentration in serum from particulate matter to Serum 47.3(μg/L). Absorption concentration of nickel in serum 16.5(μg/L) was lower than lead observed in worker's population. Absorption concentration of cadmium and chromium in serum from particulate matter -152(μg/L) observed very low. The results also show the need for immediate improvements in workplace ventilation and industrial hygiene practices.

Background: Dengue virus is the causative agent of dengue fever, a vector borne infection which causes self-limiting to life threatening disease in humans. A sero-epidemiological study was conducted to understand the current epidemiology of dengue virus in Pakistan which is now known as a dengue endemic country after its first reported outbreak in 1994., Methods: To investigate the prevalence of dengue virus in Pakistan during 2009-2014, a total of 9,493 blood samples were screened for the detection of anti-dengue IgM antibodies using ELISA. Clinical and demographic features available with hospital records were reviewed to ascertain mortalities related to dengue hemorrhagic shock syndrome., Results: Out of 9,493 samples tested, 37% (3,504) were found positive for anti-dengue IgM antibodies. Of the seropositive cases, 73.6% (2,578/3,504) were male and 26.4% (926/3,504) were female. The highest number (382/929; 41.1%) of sero-positive cases was observed among the individuals of age group 31-40 years. The highest number of symptomatic cases was reported in October (46%; 4,400/9,493), and the highest number of sero-positive cases among symptomatic cases was observed in November (45.7%; 806/1,764). Mean annual patient incidence (MAPI) during 2009-2014 in Pakistan remained 0.30 with the highest annual patient incidence (11.03) found in Islamabad. According to the available medical case record, 472 dengue related deaths were reported during 2009-2014., Conclusion: The data from earlier reports in Pakistan described the dengue virus incidence from limited areas of the country. Our findings are important considering the testing of clinical samples at a larger scale covering patients of vast geographical regions and warrants timely implementation of dengue vector surveillance and control programs., Trial Registration Number: It is an epidemiological research study, so trial registration is not required.

Objectives: Widespread implementation of electronic databases has improved the accessibility of plaintext clinical information for supplementary use. Numerous machine learning techniques, such as supervised machine learning approaches or ontology-based approaches, have been employed to obtain useful information from plaintext clinical data. This study proposes an automatic multi-class classification system to predict accident-related causes of death from plaintext autopsy reports through expert-driven feature selection with supervised automatic text classification decision models., Methods: Accident-related autopsy reports were obtained from one of the largest hospital in Kuala Lumpur. These reports belong to nine different accident-related causes of death. Master feature vector was prepared by extracting features from the collected autopsy reports by using unigram with lexical categorization. This master feature vector was used to detect cause of death [according to internal classification of disease version 10 (ICD-10) classification system] through five automated feature selection schemes, proposed expert-driven approach, five subset sizes of features, and five machine learning classifiers. Model performance was evaluated using precisionM, recallM, F-measureM, accuracy, and area under ROC curve. Four baselines were used to compare the results with the proposed system., Results: Random forest and J48 decision models parameterized using expert-driven feature selection yielded the highest evaluation measure approaching (85% to 90%) for most metrics by using a feature subset size of 30. The proposed system also showed approximately 14% to 16% improvement in the overall accuracy compared with the existing techniques and four baselines., Conclusion: The proposed system is feasible and practical to use for automatic classification of ICD-10-related cause of death from autopsy reports. The proposed system assists pathologists to accurately and rapidly determine underlying cause of death based on autopsy findings. Furthermore, the proposed expert-driven feature selection approach and the findings are generally applicable to other kinds of plaintext clinical reports., Competing Interests: The authors have declared that no competing interests exist.

Objective: To evaluate NS1 antigen detection ELISA for the early laboratory diagnosis of dengue virus infection., Methods: The present study was conducted to evaluate the overall positivity of NS1 antigen detection ELISA and its comparison with viral RNA detection via real time PCR and IgM antibodies detection by ELISA., Results: A total of 1270 serum samples were tested 86% (1097/1270) were detected positive by one or more than one diagnostic test. Out of 1 270, 64% (807/1270) were positive by NS1 ELISA and 52% (662/1270), 51% (646/1270) were positive by real-time RT-PCR and IgM ELISA respectively., Conclusions: NS1 antigen detection ELISA is highly suitable diagnostic tools and it also has great value for use in outbreak and epidemic situation., (Copyright © 2016 Hainan Medical University. Production and hosting by Elsevier B.V. All rights reserved.)

The current study explored the potential links between breast cancer and human interferon α-2b (hIFNα-2b) gene mutations. The hIFNα-2b gene was amplified from breast cancer tumor tissue samples (N = 60) by polymerase chain reaction (PCR) and the products were subjected to gene sequencing. A total of 38 (63.3%) samples showed positive PCR amplification results. Several of these also exhibited frequent alterations (mutations) after 400 bp and, in particular, adenine was replaced by other bases. A total of 19 selected mutated amino acids were analyzed for local/general fold pattern changes. Human IFNα-2b receptor (IFNAR): ligand (hIFNα-2b protein) interactions through a Z-DOCK (3.0.2) server were also evaluated to assess the binding patterns of each ligand to receptor to induce Janus-Kinase-signal transducer and activator of transcription antiproliferative signal transduction pathway inside the cancer cells. Certain local structural and conformational changes were predicted to be induced by mutations in the ligand. The variant models of the hIFNα-2b displayed structural and conformational changes that signified that changes to hIFNα-2b may be a risk factor in addition to other known factors associated with onset/progression of female breast carcinoma. It was hoped that others might build upon the research in this study evaluating protein structural models with mutations and their consequent interactions with receptors in the development of potent immune therapeutic drugs for breast cancer that are based on recombinant hIFNα-2b.

Objective: To determine the prevalence of cytomegalovirus in pregnant women and types of overt congenital infection in neonates., Methods: This cross-sectional study was conducted at the Pakistan Institute of Medical Sciences and Federal Government Services Hospital in Islamabad, Pakistan, from March 2010 to June 2011, and comprised blood samples of pregnant women. Seroprevalence of human cytomegalovirus, immunoglobulin G and immunoglobulin M was determined by enzyme-linked immunosorbent assay while its deoxyribonucleic acid was detected by nested polymerase chain reaction.The congenital human cytomegalovirus infection was also identified in newborn babies from actively infected pregnant women. SPSS 18 was used for data analysis., Results: Of the 409 pregnant women enrolled, 399(97.55%) were seropositive for cytomegalovirus immunoglobulinG and 52(12.71%) for immunoglobulinM, while cytomegalovirus deoxyribonucleic acid was detected in 82(20%). Of the cytomegalovirus immunoglobulinM-positive women, sera of 40(80%) had immunoglobulinG avidity >50%. The remaining 12(23%) sera had avidity assay value <50%. Among the 82(20%) infected pregnant women, 70(85.4%) were successfully followed up. Among them, the virus was isolated from 41(58.5%) newborns babies, of which 15(21%) were symptomatic while 26(47.2%) were asymptomatic. Of the former, 4(26.6%) had hepatosplenomegaly., Conclusions: Human cytomegalovirus infection in pregnant women was the main reason of congenital defects among neonates.

Background: The popularity and proliferation of online social networks (OSNs) have created massive social interaction among users that generate an extensive amount of data. An OSN offers a unique opportunity for studying and understanding social interaction and communication among far larger populations now more than ever before. Recently, OSNs have received considerable attention as a possible tool to track a pandemic because they can provide an almost real-time surveillance system at a less costly rate than traditional surveillance systems., Methods: A systematic literature search for studies with the primary aim of using OSN to detect and track a pandemic was conducted. We conducted an electronic literature search for eligible English articles published between 2004 and 2015 using PUBMED, IEEExplore, ACM Digital Library, Google Scholar, and Web of Science. First, the articles were screened on the basis of titles and abstracts. Second, the full texts were reviewed. All included studies were subjected to quality assessment., Result: OSNs have rich information that can be utilized to develop an almost real-time pandemic surveillance system. The outcomes of OSN surveillance systems have demonstrated high correlations with the findings of official surveillance systems. However, the limitation in using OSN to track pandemic is in collecting representative data with sufficient population coverage. This challenge is related to the characteristics of OSN data. The data are dynamic, large-sized, and unstructured, thus requiring advanced algorithms and computational linguistics., Conclusions: OSN data contain significant information that can be used to track a pandemic. Different from traditional surveys and clinical reports, in which the data collection process is time consuming at costly rates, OSN data can be collected almost in real time at a cheaper cost. Additionally, the geographical and temporal information can provide exploratory analysis of spatiotemporal dynamics of infectious disease spread. However, on one hand, an OSN-based surveillance system requires comprehensive adoption, enhanced geographical identification system, and advanced algorithms and computational linguistics to eliminate its limitations and challenges. On the other hand, OSN is probably to never replace traditional surveillance, but it can offer complementary data that can work best when integrated with traditional data., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Background: Congenital cytomegalovirus (cCMV) infection contributes to considerable long-term sequelae in neonates and children all over the world. The association between viral genotypes and severity of clinical cytomegalovirus (CMV) infection is yet to be defined. The objective of this study was to find the impact of active CMV infection during pregnancy and the clinical significance of genotypes in neonates with congenital cytomegalovirus infections in Pakistan., Methods: A total of 409 blood samples from pregnant women seeking health care services at the two antenatal hospitals of Islamabad during January to December 2012 were tested by ELISA and nested-PCR. Pregnant women with active infection (detected as IgM positive, PCR positive or positive on both assays) were followed until delivery, to detect the outcome of overt cCMV infection in neonates. Genetic characterization of CMV strains was performed by sequence analysis of envelope glycoproteins: gB, gN and gH to detect the contributing CMV genotypes., Results: The seroprevalence of anti-CMV IgG and IgM was 97.5% (399 out of 409) and 12.7% (52 out of 409), respectively, while 20% (82/409) pregnant women were found positive for CMV DNA by PCR. Logistic regression analysis showed a significant association of active infection with parity [OR = 2.56, 95% CI = 1.82-2.62, p = 0.04], febrile illness [OR = 1.84, 95% CI = 1.76-3.65, p = 0.01] and jaundice [OR = 22.5, 95% CI = 4.53-85.02, p = 0.002]. We were able to isolate virus in 41 out of 70 neonates; 36.6% (15 out of 41) of them were symptomatic at birth while 63.4% (26 out of 41) were asymptomatic. The most prominent clinical feature observed in symptomatic neonates was hepatosplenomegaly (26.6%; 4 out of 15). All three genotypes gB, gN and gH were found with the highest frequency of gB1 genotype, found in 75% infants with hepatic damage. Phylogenetic analysis of Pakistani strains showed 96%-100% homology to their prototype strains., Conclusions: Active CMV infection during pregnancy is a major cause of congenital CMV infection with comparable distribution of all three genotypes: gB, gN and gH in symptomatic and asymptomatic neonates. Our findings emphasize to conduct a comprehensive large scale survey and introduction of country wide routine screening at maternity clinics for early diagnosis of CMV to reduce its associated devastating outcomes.

Objective: To high light some epidemiological, clinical and diagnostic features of dengue fever during an outbreak and the role of different diagnostic techniques to achieve the highest level of accuracy in results., Methods: Blood samples (n = 323) were collected along with epidemiological and clinical data from suspected dengue patients who visited different hospitals in Swat and Mansehra district of Pakistan between May-November 2013 during a dengue outbreak. Samples were tested for the detection of viral nucleic acid by real-time PCR, non structural protein-1 (NS1) antigen and IgM antibodies by ELISA., Results: Out of 323 cases with clinical dengue infection, 304 were positive by one or more diagnostic parameter; 201 samples were positive by real-time PCR, 209 were positive by NS1 ELISA and 190 were positive by IgM antibodies. Sensitivities of real-time PCR and NS1 ELISA were comparable for early diagnosis of dengue virus infection, IgM antibody detection assay was found useful for the diagnosis in the samples collected later than day 5 of onset., Conclusions: The use of real-time PCR or detection of non structural protein NS1 by ELISA followed by IgM antibodies detection can be recommended for early diagnosis of dengue virus infection with a high level of accuracy., (Copyright © 2016 Hainan Medical College. Production and hosting by Elsevier B.V. All rights reserved.)

Mutations of GJB2 which encode connexin 26, contribute to 6-7 % of profound deafness in Pakistan. We investigated the involvement of GJB2 mutations in a cohort of 84 pedigrees and 86 sporadic individuals with moderate or severe hearing loss. Individuals in eight consanguineous families and four sporadic cases (9.52 and 4.65 %, respectively) were homozygous or compound heterozygous for p.W24X or p.W77X mutations in GJB2. These two variants are also among the most common mutations known to cause profound deafness in South Asia. The association of identical mutations with both profound and less severe phenotype of hearing loss suggests that alleles of other genes modify the phenotype due to these GJB2 nonsense mutations. Our study demonstrates that GJB2 mutations are an important contributor to aetiology of moderate to severe hearing loss in Pakistan.

Background: Hearing loss is a heterogeneous neurosensory disorder. Mutations of 56 genes are reported to cause recessively inherited non-syndromic deafness., Objective: We sought to identify the genetic lesion causing hearing loss segregating in a large consanguineous Pakistani family., Methods and Results: Mutations of GJB2 and all other genes reported to underlie recessive deafness were ruled out as the cause of the phenotype in the affected members of the participating family. Homozygosity mapping with a dense array of one million SNP markers allowed us to map the gene for recessively inherited severe hearing loss to chromosome 7q31.2, defining a new deafness locus designated DFNB97 (maximum logarithm of the odds score of 4.8). Whole-exome sequencing revealed a novel missense mutation c.2521T>G (p.F841V) in MET (mesenchymal epithelial transition factor), which encodes the receptor for hepatocyte growth factor. The mutation cosegregated with the hearing loss phenotype in the family and was absent from 800 chromosomes of ethnically matched control individuals as well as from 136 602 chromosomes in public databases of nucleotide variants. Analyses by multiple prediction programmes indicated that p.F841V likely damages MET function., Conclusions: We identified a missense mutation of MET, encoding the hepatocyte growth factor receptor, as a likely cause of hearing loss in humans., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.)

The plant diversity of Himalayan region has been reduced to greater extent due to environmental degradation and human exploitation. Anthropogenic disturbance was the major factor responsible for fragmentation of forest vegetation into small patches. Little research has been conducted in the Himalayan region of Poonch Valley of North eastern Pakistan with reference to plants biodiversity and its conservation. The present research was carried out to provide a checklist of vegetation for biodiversity conservation. A total of 430 vascular and 5 nonvascular plant species with 5 species of Bryophytes (5 families), 13 species of Pteridophytes (6 families), 4 species of Gymnosperms (1 family) and 413 species of angiosperms (95 families) were enumerated from the Poonch valley Azad Kashmir. The genera were classified into three categories according to the number of species. 25 plant communities with phytosociological parameters and diversity indices were reported. Present study revealed that there were 145 threatened, 30 endangered, 68 vulnerable and 47 rare species. It is recorded that extensive grazing, uprooting of plants and soil slope erosion intensify the environmental problems. Since there is maximum exploitation of vegetation, the valley showed a decline in plant diversity. The study was also indicated that the main threats to the biodiversity are expansion of settlement and army installations in the forest area of the valley. For sustainable use In-situ and Ex-situ conservation, controlled harvesting and afforestation may be the solution. Moreover, forest area should be declared prohibited for settlements and army installations.

Mutations of GJB2, which encodes connexin 26, are the most common cause of hereditary hearing loss in many human populations. This study was initiated to determine the prevalence of GJB2 mutations in individuals with hearing loss from the Hazara Division in Pakistan. We recruited 70 participants with nonsyndromic deafness segregating as an apparently recessive trait and directly sequenced the GJB2 coding region from their DNA. The homozygous mutations c.71 G → A (p.W24X), c.104 T → G (p.I35S), and c.35delG (p.G12VfsX1) were identified as the cause of hearing loss in three participants (4.28%); in populations from other areas of Pakistan, frequencies of 6-7% have been observed. The mutations c.104 T → G and c.35delG were identified in Pakistan for the first time. These results confirm the low prevalence of GJB2 mutations in Hazara and suggest that mutations in other genes may play a significant role in the etiology of deafness in this population.

A two-stage process, composed of growth under nutrient-rich conditions followed by cultivation under nitrogen starvation and controlled conditions of phosphate, light intensity, aeration, and carbon sources was applied for lipid production by the green alga Chlorella vulgaris. Using conditions without addition of nitrogen, 2mg/L PO(4)-P, light intensity of 100μmol/m(2)/s and 0.25vvm of air, about 43% of dry cell weight accumulated as lipids after 12h, which equates to a lipid productivity of 77.8mg/L/d. In a medium containing 5mg/L NO(3)-N and 2mg/L PO(4)-P, and at a light intensity of 100μmol/m(2)/s and 0.25vvm of 2% CO(2), about 53% of dry cell weight consisted of lipids after 24h, representing a lipid productivity of 77.1mg/L/d. The low amount of nutrients, moderate aeration and light intensity were helpful for increasing lipid productivity., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Mutations in PJVK, encoding Pejvakin, cause autosomal recessive nonsyndromic hearing loss in humans at the DFNB59 locus on chromosome 2q31.2. Pejvakin is involved in generating auditory and neural signals in the inner ear. We have identified a consanguineous Pakistani family segregating sensorineural progressive hearing loss as a recessive trait, consistent with linkage to DFNB59. We sequenced PJVK and identified a novel missense mutation, c.1028G>C in exon 7 (p.C343S) co-segregating with the phenotype in the family. The p.C343 residue is fully conserved among orthologs from different vertebrate species. We have also determined that mutations in PJVK are not a common cause of hearing loss in families with moderate to severe hearing loss in Pakistan. This is the first report of PJVK mutation in a Pakistani family and pinpoints an important residue for PJVK function., (Copyright © 2012 Elsevier B.V. All rights reserved.)

During August to November 2010, a dengue (DEN) outbreak occurred, and several cities of Pakistan were affected. In this study, serum samples from 317 patients, based on clinical symptoms and blood test, were included from Lahore, Sheikhpura and Gujranwala cities for further diagnosis of different genotypes of DEN virus. The samples were categorized into 2 groups: group 1 containing patients with dengue fever (DF) without hemorrhagic manifestations (DF) (n=228) and group 2 involving patients having DF with hemorrhagic manifestations (dengue hemorrhagic fever) (n=89). The samples found positive for DEN in reverse transcriptase polymerase chain reaction (PCR) by targeting capsid and premembrane genes containing fragment of size ∼2.5 kb for groups 1 and 2 were found to be 83.33% and 91.01%, respectively; however, the detection limit was quite low by immunoglobulin M antibody capture enzyme linked immunosorbent assay in case of both groups and was found to be 34.70% and 68.53%, respectively. The 16.67% and 8.99% samples of groups 1 and 2, respectively, were found negative in reverse transcriptase PCR analysis. In case of group 1 and group 2, by adopting seminested multiplex PCR strategy, predominant genotype was DEN virus type 2 with prevalence rates of 45.17% and 65.16%, respectively. DEN virus type 1 was the second most prevailing genotype in the population with prevalence rates of 18.85% and 19.10% for groups 1 and 2, respectively.

Background: Dengue Fever is the most common arboviral disease in the world, and presents cyclically in tropical and subtropical regions of the world. The four serotypes of dengue virus, 1, 2, 3, and 4, form an antigenic subgroup of the flaviviruses (Group B arboviruses). Transmission to humans of any of these serotypes initiates a spectrum of host responses, from in apparent to severe and sometimes lethal infections. Complete Blood count (CBC) is an important part of the diagnostic workup of patients. Comparison of various finding in CBC including peripheral smear can help the physician in better management of the patient., Material and Methods: This cross sectional study was carried out on a series of suspected patients of Dengue viral infection reporting in Ittefaq Hospital (Trust). All were investigated for serological markers of acute infection., Results: Out of 341 acute cases 166 (48.7%) were confirmed by IgM against Dengue virus. IgG anti-dengue was used on 200 suspected re-infected patients. Seventy-one (39.5%) were positive and 118 (59%) were negative. Among 245 confirmed dengue fever patients 43 (17.6%) were considered having dengue hemorrhagic fever on the basis of lab and clinical findings. Raised haematocrit, Leukopenia with relative Lymphocytosis and presence atypical lymphocytes along with plasmacytoid cells was consistent finding at presentation in both the patterns of disease, i.e., Dengue Haemorrhagic fever (DHF) and Dengue fever (DF)., Conclusion: Changes in relative percentage of cells appear with improvement in the symptoms and recovery from the disease. These findings indicate that in the course of the disease, there are major shifts within cellular component of blood.

Objective: To ascertain the current approach to screen for developmental dysplasia of the hip in the Republic of Ireland., Methods: Two-pronged prospective and retrospective study. (1) Postal questionnaire to consultant paediatricians responsible for the routine neonatal care of infants in the Irish Republic in June 2006. (2) Retrospective database review to identify infants undergoing radiological follow-up and their outcome., Results: All maternity units surveyed responded. Most units (84%) were dependent on radiographs at 4-6 months for imaging hips, only two units primarily used ultrasound (10.5%). We estimate that neonatal hip examination is performed by an experienced examiner in less than 30% of routine newborn examinations. On retrospective analysis, 94% of radiographs performed were normal., Conclusions: The most effective interventions, selective ultrasound and examination by an experienced clinician are not widely practiced. There is a need for the development of national guidelines based on available resources.

A case of a biliopleural fistula with a biloma occurring after superselective hepatic transarterial chemoembolization ablation of a metastatic hepatic carcinoid is described. The presentation was complicated by choledocholithiasis. The biloma was successfully treated with endoscopic drainage.

Background: Psoralen Ultraviolet A (PUVA) therapy is a well-established treatment of psoriasis. The objective of the study was to compare the clinical improvement in psoriasis with PUVA photochemotherapy + topical corticosteroids and PUVA + bland emollients., Methods: Forty patients with chronic plaque type of psoriasis were divided into two equal groups each having 20 patients. PUVA therapy was given thrice weekly. In addition, patients of group-A were allowed to apply topical betamethasone 17-valerate 0.1% diluted 1 into 2 parts with plain vaseline twice daily. Patients of group-B were allowed to apply only plain vaseline over lesions twice daily. Clinical improvement in lesions was observed by decrease in the severity of erythema, scaling and plaque elevation., Results: Clearance of psoriasis was achieved in 95%, of the patients treated with PUVA plus topical corticosteroids while clearance was achieved in 80% of patients treated with PUVA plus bland emollients (P = 0.0758). Median numbers of exposures for group-A were 16 & for group-B were 17.5 (p = 0.1029). Similarly, median cumulative dose in group-A was 64.5 J/cm2 & in group-B was 70.7 J/cm2 (p = 0.372)., Conclusion: There is no significant difference in clinical improvement in psoriasis treated either by PUVA plus topical steroids or PUVA plus bland emollients.

Objective: To compare the clinical improvement in psoriasis with psoralen - UVA photochemotherapy versus narrow band UVB phototherapy alone., Design: An interventional quasi- experimental study., Place and Duration of Study: Dermatology Department, Nishtar Hospital, Multan, from May 2002 to June 2003., Patients and Methods: Forty patients with chronic plaque type psoriasis were included in the study. These were divided into two equal groups each having 20 patients. PUVA or UVB therapy was given thrice weekly. Clinical improvement in lesions was observed by decrease in the severity of erythema, scaling and plaque elevation., Results: Clearance of psoriasis was achieved in significantly greater proportion of patients treated with PUVA (85%) than with UVB phototherapy (60%) (p=0.03814). Significantly fewer number of exposures were required for clearance with PUVA (median number of treatments=17) as compared to UVB (median number of treatments=25.5) (p=0.0002). Median cumulative dose for PUVA was 69.1 J/cm2 and for UVB was 34.15 J/cm2, respectively (p<0.0001)., Conclusion: When given thrice weekly, PUVA was more effective treatment for psoriasis than narrow band UVB phototherapy.

Objective: To identify the Leishmania species being responsible for cutaneous leishmaniasis in Multan., Method: Parasites were isolated from clinically and parasitologically confirmed lesions of cutaneous leishmaniasis from 30 patients by fine needle aspiration (FNA). The bioptical materials were then cultured in Evans Tobie's medium and parasites isolated were identified by isoenzyme electrophoresis technique., Results: Successful Leishmania isolates were obtained from 16 patients. All strains were identified by biochemical techniques as belonging to Leishmania tropica zimodeme MON7 variant PGD., Conclusion: The causative species was identified as Leishmania tropica.

Objective: Cutaneous leishmaniasis is endemic in Multan. Therefore this study was undertaken to elucidate the epidemiological features with main emphasis on the geographical distribution of the infection in Multan., Settings: Multan city. Dermatology Outpatient Department, Nishtar Medical College., Method: Records of 173 diagnosed patients of cutaneous leishmaniasis were retrospectively reviewed according to the age, sex, number and site of lesions, results of the smear for LD bodies and residential area., Results: The disease was more common in the children and young adults, with male to female ratio of 3:2. The maximum number of lesions were on the lower legs. Sixty five percent of the patients were from the interior old city., Conclusion: The main focus of infection is in old interior part of the city with microfoci of the disease scattered throughout the city.

Background: Cutaneous leishmaniasis (CL) is endemic in certain areas of Pakistan. This study was carried out to determine the incidence and clinical pattern of the disease in Pakistan., Methods: CL patients presenting in the out-patient clinic of the Department of Dermatology, Nishtar Hospital, Multan from January 1995 to June 1997 were included in the study. The patients were diagnosed clinically and confirmed by laboratory demonstration of the parasite in a Giemsa-stained smear from the lesion. All important clinical details were recorded on specially designed forms., Results: Three hundred and five patients with 681 lesions were seen during 30 months. All the lesions were of the dry type. Most of the lesions (97%) were present on exposed areas of the body. One hundred and thirty-four (44%) patients had one and 75 (24%) had two lesions. Eight five per cent of the sufferers were less than 30 years of age. The disease was slightly more preponderant in males (60%). Most cases presented during winter., Conclusions: Lack of wet type lesions may be due to complete absence of Leishmania major in this area. This is in contrast to other areas of Pakistan where both dry and wet types of CL are present.