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Background and Objectives: Hereditary multiple exostoses (HME) is a disease characterized by cartilage-capped bony protuberances at the site of growth plates of long bones. Functional mutations in the exostosin genes (EXT1 and EXT2) are reported to affect the hedgehog signalling pathways leading to multiple enchondromatosis. However, the exact role of each EXT protein in the regulation of heparan sulphate (HS) chain elongation is still an enigma. In this study, a Pakistani family with HME is investigated to find out the genetic basis of the disease. Materials and Methods: Genotyping of eight members of the family by amplifying microsatellite markers, tightly linked to the EXT1 and EXT2 genes. Results: The study revealed linkage of the HME family to the EXT1 locus 8q24.1. Sanger sequencing identified a heterozygous deletion (c.247Cdel) in exon 1 of EXT1, segregating with the disease phenotype in the family. In silico analysis predicted a shift in the frame causing an early stop codon (p.R83GfsX52). The predicted dwarf protein constituting 134 amino acids was functionally aberrant with a complete loss of the catalytic domain at the C-terminus. Interestingly, an alternative open reading frame 3 (ORF3) caused by the frame shift is predicted to encode a protein sequence, identical to the wild type and containing the catalytic domain, but lacking the first 100 amino acids of the wild-type EXT1 protein. Conclusion: Consequently, haploinsufficiency could be the cause of HME in the investigated family as the mutated copy of EXT1 is ineffective for EXT-1/2 complex formation. The predicted ORF3 protein could be of great significance in understanding several aspects of HME pathogenesis. [ABSTRACT FROM AUTHOR]

Abstract Diabetes mellitus is associated with an increase in sialic acid concentration along with other complications. Sialic acid changes in NIDDM patients were investigated following bitter melon (55 ml/24 h) [...]

Abstract Background Hepatitis C virus (HCV) infection is one of the leading causes of chronic liver disease (CLD). About 80% of those exposed to the virus develop a chronic infection. Hyperhomocysteinemia, which is an independent risk factor for atherosclerotic vascular disease and thromboembolism, may develop in HCV-infected patients although altered alanine amino transferase (ALT) enzyme levels are generally associated with damage to liver cells. The gold standard therapy for chronic hepatitis C patients is pegylated interferon combined with an anti-viral drug (ribavirin). The current study aimed to investigate the effect of antiviral therapy on plasma homocysteine (Hcy) levels in HCV patients in addition to other parameters. Methods 532 HCV-infected patients and 70 healthy controls were recruited for the study. All patients were subjected to laboratory investigations including HCV-RNA levels, complete blood cell counts, serum levels of homocysteine, ALT, alkaline phosphatase (ALP), lipid profile and liver ultrasonographic examination. The outcome of treatment with pegylated interferon α plus ribavirin treatment and sustained virologic response (SVR) was determined 6–9 months post-therapy. Results Hyperhomocysteinemia was found in 91.35% of HCV-infected patients. The difference in plasma Hcy concentrations reached statistical significance between the patient and control groups. ALT, cholesterol and triglycerides (TGs) levels were found higher than normal in the patients group. After receiving a combined therapy for 24 weeks, 43.66% patients showed an SVR (responders); 30.98% patients were non-responders while 25.35% patients initially responded to therapy but again retrieved positive status of HCV infection six months post-therapy (relapse-cirrhotic patients). The mean levels of plasma Hcy, ALT and ALP were significantly reduced in responders within 10 weeks of therapy when compared with non-responders and relapse-cirrhotic patients. Conclusion Elevated homocysteine levels in serum due to HCV infection can be reduced to normal range with the standard interferon α plus ribavirin treatment. This study highlights the significance of the measurement of serum homocysteine levels in the diagnosis and monitoring of HCV infection treatment in addition to other laboratory parameters.

Non‐bullous congenital ichthyosiform erythroderma (NCIE) is characterized by skin scaling with erythema. In this study, two Pakistani families with NCIE are genetically characterized through Whole Exome and Sanger sequencing to identify molecular basis of the disease. We identified a nonsense homozygous c.2026C>T mutation of ALOXE3, causing premature termination of the eLOX3 protein (p.Q676X). In silico studies predicted impaired enzymatic activity of the premature truncated eLOX3, leading to abnormal synthesis of specific hepoxilin derivatives, essential for epidermal barrier formation. It is the first ever study reporting homozygotes of p.Q676X mutation in ethnically distinct two Pakistani families; otherwise, heterozygotes of the said mutation have been reported in South Asian population only. Hence, mutation seems to be region‐specific and may be useful for molecular diagnosis of NCIE. Moreover, our findings should help in genetic counseling and career screening. [ABSTRACT FROM AUTHOR]

To investigate the antitumor activity, brine shrimp lethality assay, antibacterial and antifungal activity of Methanol Extract (ME), Water Extract (WE), Acetone Extract (AE), Chloroform Extract (CE), Methanol-Water Extract (MWE), Methanol-Acetone Extract (MAE), Methanol-Chloroform Extract (MCE) of Ranunculus arvensis (L.). Antitumor activity was evaluated with Agrobacterium tumefaciens (At10) induced potato disc assay. Cytotoxicity was evaluated with brine shrimp lethality assay. Antibacterial activity was evaluated with six bacterial strains including Escherichia coli, Enterobacter aerogenes, Bordetella bronchiseptica, Klebsiella pneumoniae, Micrococcus luteus and Streptococcus anginosus and antifungal screening was done against five fungal strains including Aspergillus niger, A. flavus, A. fumigates, Fusarium solani and Mucor species by using disc diffusion method. Best antitumor activity was obtained with ME and WE, having highest IC50 values 20.27±1.62 and 93.01±1.33µg/disc. Brine shrimp lethality assay showed LC50 values of AE, MAE and ME were obtained as 384.66±9.42µg/ml, 724.11±8.01 µg/ml and 978.7±8.01µg/ml respectively. WE of R. arvensis revealed weak antimicrobial result against the tested microorganisms. On the other hand, the antifungal activity of the plant extracts was found to be insignificant. These findings demonstrate that extracts of R. arvensis possesses significant antitumor activity. Further extensive study is necessary to assess the therapeutic potential of the plant. [ABSTRACT FROM AUTHOR]

Purpose: To determine the prevalence and genetic basis of tetracycline resistance in Staphylococcus aureus. Methods: One hundred and thirty (130) clinical isolates of S. aureus were collected from Khyber Teaching Hospital, Peshawar, Pakistan. Susceptibility to antibiotics (doxycycline, tetracycline and minocycline) was determined by Kirby-Bauer disc diffusion method with minimum inhibitory concentration (MIC) evaluated on Muller-Hinton agar as described by the Clinical and Laboratory Standards Institute (CLSI). The tetracyclineresistant strains (TET-R) were screened by polymerase chain reaction (PCR) for the presence of four common tetracycline resistance determinants, viz, tet(K), tet(L), tet(M) and tet(O). Results: Sixty (46.0 %) of these isolates were methicillin-resistant S. aureus (MRSA) while 70 (54.0 %) were methicillin-susceptible S. aureus (MSSA). Seventy four (56.9 %) strains were resistant to tetracycline (TET-R), 30 (23.1 %) to minocycline and 23 (17.7 %) to doxycycline. A majority of the MRSA were resistant to tetracyclines and all the MSSA were sensitive to doxycycline and minocycline. The tet(K) gene was found in 58 isolates and tet(L) in one isolate. No tet(M) and tet(O) were detected. Conclusion: This study indicates that resistance to tetracyclines is mainly by efflux pumps mediated by tet(K) in S. aureus in northwestern Pakistan. [ABSTRACT FROM AUTHOR]

Background Malaria has long been known to cause changes in biochemical parameters in blood but there is limited data on the subject from Pakistan. Therefore in this study we aimed to assess the prevalence of slide positivity for malaria and evaluate associated changes in blood chemistry in adults among residents of district Bannu. Methodology Blood samples from 3500 patients who presented to a hospital in Bannu for suspected malaria were collected between August to September 2007. The blood films of these patients were examined by Geimsa staining. Biochemical analyses (serum glucose, bilirubin, AST, ALT and creatinine levels) were performed on all samples that tested positive for malaria and compared to randomly selected samples that were negative for malaria. Results The slide positivity rate for malaria was 22% (767/3500). Among them 55 (7.2%) were Plasmodium falciparum, while 712 (92.8%) were Plasmodium vivax. A sub-sample of 70 patients was selected from the 767 malaria positive cases and compared to 40 healthy controls matching for locality and age to compare the blood biochemistry among normal and infected subjects. Patients with either P. vivax or P. falciparum had higher mean serum bilirubin (3.07mg/dl [P<0.01] and 3.9mg/dl [P=<0.001 respectively versus 0.567mg/dl in normal control subjects) and creatinine (1.07 mg/dl [P=0.0003] and 1.20 mg/dl [P=0.0003] respectively versus 0.50 mg/dl in normal control subjects). No significant change was observed in AST, ALT and glucose concentration for either P. vivax or P. falciparum. Conclusion There is a higher burden of P. vivax malaria in Bannu district. Both P. vivax and P. falciparum infections are associated with an increase in mean bilirubin and creatinine levels. [ABSTRACT FROM AUTHOR]

Examines the application of pulsed corona discharges in toxic volatile organic compounds abatement. Development of thermal and cold plasmas; Advantages of pulsed corona discharges; Prediction of the behavior of VOC in non-equilibrium plasma environment.

Background aims. Mesenchymal stromal cell (MSC) transplantation has immerged as promising therapeutic approach to treat spinal cord injury (SCI). In this pilot study, we investigated the safety of intrathecal injection of autologous bone marrowderived MSCs in nine patients with SCI. Methods. Patients with complete SCI at the thoracic level were divided into two groups: chronic (>6 months, group 1) and sub-acute SCI (<6 months, group 2), according to time elapsed since injury. MSCs were isolated by density gradient separation of autologous bone marrow harvested from the iliac crest. Cells were cultured in a Good Manufacturing Practice-compliant facility to produce clinical scale dose. After quality control testing, MSCs were injected back to patients by intrathecal injection. Safety was defined as absence of adverse event and side effects after 1 month after receiving the injection. Results. Six patients had chronic SCI with a median duration of 33 months since date of injury (range: 10-55 months), and three patients were in sub-acute phase of disease. Each patient received two or three injections with a median of 1.2 x 106 MSCs/kg body weight. No treatment-related adverse event was observed during median follow-up of 720 days (range: 630-826 days) in group 1 and 366 days (range: 269-367 days) in group 2, respectively. Discussion. This pilot study demonstrated that autologous MSCs can be safely administered through intrathecal injection in spinal cord injury patients. Further investigation through randomized, placebo-controlled trials is needed. [ABSTRACT FROM AUTHOR]

The present study was planned to carry out a seroprevalence study of PPR antibodies in North West Frontier Province (NWFP) including Federally Administered Tribal Areas (FATA), using competitive-ELISA (c-ELISA) kit. A total of 4548 serum samples collected from 26 locations of the NWFP including. Eight hundred twenty eight samples were found positive for PPR antibodies yielding an overall seroprevalence of 18.20 percent in the small ruminant population included in the study. 337 (24.90%) serum samples from sheep and 491 (15.36%) from goats were found positive in PPR c-ELISA yielding a marginal difference in seroprevalence of PPR. The current situation of PPR in Pakistan could be regarded as of low magnitude but constant threat. Incidence may increase to the alarming level due to uncontrolled/ free movement within and from across boarders to the stocks of non-immunized susceptible population of sheep and goats in the country but production systems and economic realities constraint the effective animal movement control within and across the boarders. Based on the limited reports of outbreaks and uneven distribution of occurrence over the years of past decade, the best available options are high level of prevention and objective surveillance. Keeping in view the high risk of PPR, control strategy should be adopted and further study needs to be undertaken to ascertain the extent of PPR virus circulating in the population of goats and sheep in the country. Under existing production and marketing system, sanitation and preventive vaccination are two practicable options for control of PPR in Pakistan. Homologous PPR vaccine is now available which is a freeze dried live vaccine derived from tissue culture from attenuated PPR virus. The vaccine is intended for sheep and goats and is required to be injected to all animals above the age of 3 month. The vaccine may be repeated after one year of the primary vaccination in high risk areas. [ABSTRACT FROM AUTHOR]