Mass spectrometry as a tool for the selective profiling of destruxins; their first identification in Lecanicillium longisporum.

Mass spectrometry was applied to the identification of the destruxins (dtxs), cyclic peptides that are commonly produced by the fungal insect-pathogen, Metarhizium anisopliae. The aim of the study was to optimise a methodology in order to firstly determine whether these compounds were present in other species and to determine the effect of differing growth conditions upon the dtx content detected. Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-ToF-MS) was initially used to analyse the dtxs, but limitations were indicated.

Seroprevalence of hepatitis B and C infections among young adult males in Pakistan.

This descriptive, cross-sectional study was conducted at 2 military hospitals in Pakistan from January 2004 to December 2005. Young adult males with age range 17-22 years from different districts of Pakistan were screened for hepatitis B surface antigens (HBsAg) and anti-hepatitis C antibodies (anti-HCV). Out of 5707 young men tested, 95 (1.

Spinal cord compression: to biopsy, or not to biopsy?

A patient presenting with acute spinal cord compression initially thought to be secondary to metastatic cancer based on magnetic resonance imaging alone is described. No primary tumour was identified on further imaging, and although deemed to be technically very difficult, surgical decompression was postponed in favour of obtaining a histological diagnosis. Histology confirmed a low grade non-Hodgkin's lymphoma, a diagnosis which would not have been considered had a biopsy not been undertaken.

SUMO fusion technology for enhanced protein production in prokaryotic and eukaryotic expression systems.

In eukaryotic cells, the reversible attachment of small ubiquitin-like modifier (SUMO) protein is a post-translational modification that has been demonstrated to play an important role in various cellular processes. Moreover, it has been found that SUMO as an N-terminal fusion partner enhances functional protein production in prokaryotic and eukaryotic expression systems, based upon significantly improved protein stability and solubility. Following the expression and purification of the fusion protein, the SUMO-tag can be cleaved by specific (SUMO) proteases via their endopeptidase activity in vitro to generate the desired N-terminus of the released protein partner.

Morgagni hernia presenting with bowel obstruction in a lung transplant recipient: case report.

Morgagni hernias are uncommon congenital diaphragmatic deficiencies that may remain asymptomatic till adulthood. We report a case of Morgagni hernia presenting with subacute bowel obstruction in a bilateral lung transplant recipient. This diaphragmatic deficiency was not evident during bilateral lung transplantation surgery via clamshell incision.

Engineered Saccharomyces cerevisiae strain for improved xylose utilization with a three-plasmid SUMO yeast expression system.

A three-plasmid yeast expression system utilizing the portable small ubiquitin-like modifier (SUMO) vector set combined with the efficient endogenous yeast protease Ulp1 was developed for production of large amounts of soluble functional protein in Saccharomyces cerevisiae. Each vector has a different selectable marker (URA, TRP, or LEU), and the system provides high expression levels of three different proteins simultaneously. This system was integrated into the protocols on a fully automated plasmid-based robotic platform to screen engineered strains of S.

Strategies for the identification of novel inhibitors of deubiquitinating enzymes.

Dysregulation of the UPS (ubiquitin-proteasome system) has been implicated in a wide range of pathologies including cancer, neurodegeneration and viral infection. Inhibiting the proteasome has been shown to be an effective therapeutic strategy in humans; yet toxicity with this target remains high. DUBs (deubiquitinating enzymes) represent an alternative target in the UPS with low predicted toxicity.

Risk factors for hypoxic ischemic encephalopathy in children.

Objective: To determine underlying risk factors in neonates with hypoxic ischemic encephalopathy.

Study Design: Case-control study.

Place And Duration Of Study: Neonatology Unit at the Children's Hospital and the Institute of Childhealth, Lahore, from March to July 2005.

Enhanced protein expression in the baculovirus/insect cell system using engineered SUMO fusions.

Recombinant protein expression in insect cells varies greatly from protein to protein. A fusion tag that is not only a tool for detection and purification, but also enhances expression and/or solubility would greatly facilitate both structure/function studies and therapeutic protein production. We have shown that fusion of SUMO (small ubiquitin-related modifier) to several test proteins leads to enhanced expression levels in Escherichia coli.

Enhanced protein expression in mammalian cells using engineered SUMO fusions: secreted phospholipase A2.

SUMOylation, the covalent attachment of SUMO (small ubiquitin-like modifier), is a eukaryotic post-translational event that has been demonstrated to play a critical role in several biological processes. When used as an N-terminal tag or fusion partner, SUMO has been shown to enhance functional protein production significantly by improving folding, solubility, and stability. We have engineered several SUMOs and, through their fusion, developed a system for enhancing the expression and secretion of complex proteins.

Lycotoxin-1 insecticidal peptide optimized by amino acid scanning mutagenesis and expressed as a coproduct in an ethanologenic Saccharomyces cerevisiae strain.

New methods of safe biological pest control are required as a result of evolution of insect resistance to current biopesticides. Yeast strains being developed for conversion of cellulosic biomass to ethanol are potential host systems for expression of commercially valuable peptides, such as bioinsecticides, to increase the cost-effectiveness of the process. Spider venom is one of many potential sources of novel insect-specific peptide toxins.

Bone marrow transplant cure for beta-thalassaemia major: initial experience from a developing country.

Between July 2001 and June 2007, 48 consecutive patients with beta-thalassaemia major received allogeneic haematopoietic stem cell transplants (allo HSCT) from human-leukocyte-antigen-matched siblings at the Armed Forces Bone Marrow Transplant Centre, Rawalpindi, Pakistan, using standard conditioning regimens. The median age of the patient cohort was 4 years (range, 1-14 years). Thirty-one patients were in risk class I, 11 in class II and six patients were in class III.

Characterization of ubiquitin and ubiquitin-like-protein isopeptidase activities.

Conjugation or deconjugation of ubiquitin (Ub) or ubiquitin-like proteins (UBLs) to or from cellular proteins is a multifaceted and universal means of regulating cellular physiology, controlling the lifetime, localization, and activity of many critical proteins. Deconjugation of Ub or UBL from proteins is performed by a class of proteases called isopeptidases. Herein is described a readily quantifiable novel isopeptidase assay platform consisting of Ub or UBL fused to the reporter enzyme phospholipase A(2) (PLA(2)).

Investigation of ecological and environmental determinants for the presence of questing Ixodes ricinus (Acari: Ixodidae) on Gower, South Wales.

The spatial heterogeneity of questing Ixodes ricinus (L.) (Acari: Ixodidae) within endemic areas in Great Britain is well established. Their presence is acutely responsive to blood host availability and their ability to maintain water balance, which are in turn governed by a variety of ecological and environmental factors.

Risk assessment of landfill disposal sites--State of the art.

A risk assessment process can assist in drawing a cost-effective compromise between economic and environmental costs, thereby assuring that the philosophy of 'sustainable development' is adhered to. Nowadays risk analysis is in wide use to effectively manage environmental issues. Risk assessment is also applied to other subjects including health and safety, food, finance, ecology and epidemiology.

Repeated in vitro subculturing alters spore surface properties and virulence of Metarhizium anisopliae.

Repeated subculturing caused rapid changes in the spore surface properties and virulence of Metarhizium anisopliae. Of the two strains evaluated, M. anisopliae V245 attenuated more rapidly than V275.

Localization of autosomal recessive congenital cataracts in consanguineous Pakistani families to a new locus on chromosome 1p.

Purpose: To identify the disease locus for autosomal recessive congenital cataracts in two consanguineous Pakistani families.

Methods: Two Pakistani families were ascertained, ophthalmologic examination including slit lamp biomicroscopy was performed on all members, blood samples were collected and DNA was extracted. A genome-wide scan was performed using 382 polymorphic microsatellite markers on genomic DNA from affected and unaffected family members.

Justifying the clinical use of fresh frozen plasma - an audit.

Objective: To determine the appropriateness of fresh frozen plasma (FFP), uses in various haematological and clinical disorders, with reference to the British Committee for Standards in Haematology (BCSH) guidelines through an audit.

Design: Descriptive study.

Place And Duration Of Study: Department of Haematology and Transfusion Medicine, Shalamar Hospital, Lahore, from June 2001 to June 2004.

Deubiquitinating enzymes as novel anticancer targets.

Tagging proteins with mono- or poly-ubiquitin is now recognized as a multifaceted and universal means of regulating cell growth and physiology. It does so by controlling the cellular lifetime of nearly all eukaryotic proteins and the cellular localization of many critical proteins. Enzymes of the ubiquitin pathway add (ligases) or remove (deubiquitinases [DUBs]) ubiquitin tags to or from their target proteins in a selective fashion.

Effect of nutrition on growth and virulence of the entomopathogenic fungus Beauveria bassiana.

Three isolates of the entomopathogen Beauveria bassiana along with one strain of Metarhizium anisopliae were cultured on seven media with different carbon/nitrogen (C/N) ratios. The effect of nutrition on virulence of the isolates was evaluated via measurement of colony growth, spore yield, germination speed, conidial C/N ratio and Pr1 (a serine protease) activity. 'Osmotic stress' medium produced the lowest colony growth with low numbers of conidia in all isolates.

Dual detection of fungal infections in Drosophila via recognition of glucans and sensing of virulence factors.

The Drosophila immune system discriminates between various types of infections and activates appropriate signal transduction pathways to combat the invading microorganisms. The Toll pathway is required for the host response against fungal and most Gram-positive bacterial infections. The sensing of Gram-positive bacteria is mediated by the pattern recognition receptors PGRP-SA and GNBP1 that cooperate to detect the presence of infections in the host.

Peripheral blood-based polymerase chain reaction in diagnosis of pulmonary tuberculosis.

Background: The rapid diagnosis of infectious diseases, particularly those that represent a public health problem, like tuberculosis, is a challenging problem. By using nucleic acid amplification techniques like PCR, one may be able to diagnose, the disease on the day of arrival of specimen in the laboratory. For diagnosis of tuberculosis by direct methods like PCR, specimens from site of infection are required.

Changing trends in drug resistance among typhoid salmonellae in Rawalpindi, Pakistan.

We analysed the record of blood cultures carried out at the Armed Forces Institute of Pathology, Rawalpindi between 1996 and 2003. We isolated 477 Salmonella typhi and 410 S. paratyphi A from blood of patients suffering from typhoid fever.

Validity of nalidixic acid screening in fluoroquinolone-resistant typhoid salmonellae.

Objective: To validate the screening of low-level fluoroquinolone resistance in typhoid salmonellae by using nalidixic acid (30 mg) disk providing an acceptable zone of inhibition.

Design: Quasi-experimental study.

Place And Duration Of Study: The Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, Pakistan from July 2002 to June 2003.