Measurement of radioactivity and heavy metal levels in edible vegetables and their impact on Kuala Selangor communities of Peninsular Malaysia.

Vegetable is an essential daily diet item for the people of Malaysia. This work addressed the radiation and heavy metal exposure scenarios through the consumption of vegetables. Kuala Selangor is located in Sungai Selangor estuary in the west coast of Peninsular Malaysia, which is susceptible to pollution load due to the presence of large-scale industrial and human activities. Radioactivity and heavy metals level in human diet is of particular concern for the assessment of possible radiological and chemical hazards to human health. Therefore, a comprehensive study was carried out to determine the radioactivity levels ((226)Ra, (228)Ra and (40)K) and heavy metal concentrations (Cr, As, Cd, Mn, Mg, Al, Sr, Rb, Sb, Ba, Hg, Fe, Ni, Zn, Cu, Bi and Pb) in 10 varieties of vegetable collected from different farmlands in Kuala Selangor region. The committed doses for (226)Ra, (228)Ra and (40)K due to consumption of vegetables were found 16.6±1.3, 23.6±1.7 and 58±5 µSv y(-1), respectively, with a total of 98±8 µSv y(-1). This dose imposes no significant threat to human health. The estimated cancer risk shows that probability of increase in cancer risk from daily intake of vegetables is only a minor fraction of International Commission on Radiological Protection values. The concentrations of heavy metal were below the daily intake recommended by the international organisations.

Spectral estimation of nonstationary EEG using particle filtering with application to event-related desynchronization (ERD).

This paper proposes non-Gaussian models for parametric spectral estimation with application to event-related desynchronization (ERD) estimation of nonstationary EEG. Existing approaches for time-varying spectral estimation use time-varying autoregressive (TVAR) state-space models with Gaussian state noise. The parameter estimation is solved by a conventional Kalman filtering. This study uses non-Gaussian state noise to model autoregressive (AR) parameter variation with estimation by a Monte Carlo particle filter (PF). Use of non-Gaussian noise such as heavy-tailed distribution is motivated by its ability to track abrupt and smooth AR parameter changes, which are inadequately modeled by Gaussian models. Thus, more accurate spectral estimates and better ERD tracking can be obtained. This study further proposes a non-Gaussian state space formulation of time-varying autoregressive moving average (TVARMA) models to improve the spectral estimation. Simulation on TVAR process with abrupt parameter variation shows superior tracking performance of non-Gaussian models. Evaluation on motor-imagery EEG data shows that the non-Gaussian models provide more accurate detection of abrupt changes in alpha rhythm ERD. Among the proposed non-Gaussian models, TVARMA shows better spectral representations while maintaining reasonable good ERD tracking performance.

Construction and characterization of an auxotrophic ctxA mutant of O139 Vibrio cholerae.

Cholera caused by the O139 serogroup still remains a public health concern in certain regions of the world and the existing O1 vaccines do not cross-protect cholera caused by this serogroup. An aminolevulinic acid (ALA) auxotroph vaccine candidate against the O139 serogroup, designated as VCUSM2, was recently developed. It was found to be immunogenic in animal model studies but showed mild reactogenic effects due to the presence of two intact copies of Vibrio cholerae toxin (CTX) genetic element. In the present study we have modified the ctx operon by systematic allelic replacement methodology to produce a mutant strain, designated as VCUSM14. This strain has two copies of chromosomally integrated and mutated ctxA gene, encoding immunogenic but not toxic cholera toxin A subunit (CT-A). The amino acids arginine and glutamic acid at position 7th and 112th, respectively, in CT-A of VCUSM14 were substituted with lysine (R7K) and glutamine (E112Q), respectively. Two copies of the ace and zot genes present in the ctx operon were also deleted. Cholera toxin-ELISA using GM1 ganglioside showed that the both wild type CT and mutated CT were recognized by anti-CT polyclonal antibodies. VCUSM14 produced comparatively less amount of antigenic cholera toxin when compared to the VCUSM2 and Bengal wild type strain. VCUSM14 did not elicit fluid accumulation when inoculated into rabbit ileal loops at doses of 10(6) and 10(8) CFU. The colonization efficiency of VCUSM14 was one log lower than the parent strain, VCUSM2, which can be attributed to the ALA auxotrophy and less invasive properties of VCUSM14. VCUSM14, thus a non-reactogenic auxotrophic vaccine candidate against infection by O139 V. cholerae.

Breast and Axillary Lymph Nodes Metastasis five years after Radical Nephrectomy for Renal Cell Carcinoma A Case Report and Review of the Literature.

Breast is an uncommon and rare site for metastasis. Primary and secondary tumors of the breast need to be differentiated as management is different. We present a 67 year old female patient with two breast lumps and an axillary lymph node, 5 years after nephrectomy for Renal Cell Carcinoma (RCC). Mammogram report showed a dense spiculated mass at right upper outer quadrant and a retroareolar mass associated with clustered micro calcifications. Fine Needle Cytology and trucut biopsy were inconclusive. Computer Topography (CT) abdomen did not show evidence of RCC recurrence. After discussion with the patient, she underwent mastectomy with axillary clearance and the final histopathology report was consistent with metastasis from RCC. The management of this case is discussed.

Work and peak torque during eccentric exercise do not predict changes in markers of muscle damage.

OBJECTIVES: Large inter-subject variability in responses to eccentric exercise has been reported. This study investigated the hypothesis that the variability of changes in indirect markers of exercise-induced muscle damage (EIMD) would be explained by work performed and/or torque generated during eccentric exercise. METHODS: Subjects (n = 53) performed 60 maximal eccentric actions of the elbow flexors on an isokinetic dynamometer that forcibly extended the elbow joint from 60 degrees to 180 degrees at a constant velocity (90 degrees s(-1)). Markers of EIMD included maximal voluntary isometric contraction torque at 90 degrees elbow flexion (MVC), range of motion, plasma creatine kinase activity and muscle soreness. Measurements were taken 2 days before, immediately after and 1-4 days post-exercise. Pearson's correlation coefficient was used to examine relationships between exercise parameters (total work, change in total work, torque produced during exercise, change in peak torque) and markers of EIMD. RESULTS: Large inter-subject variability was evident for both work and torque during exercise, and changes in all markers of EIMD. Contrary to the hypothesis, total work (normalised for individual pre-exercise MVC) did not correlate significantly with any markers of EIMD, with the exception of MVC (r = 0.3). Total work performed and changes in total work showed higher correlations with some markers, but no r-values exceeded 0.4. Normalised exercise torque and the changes in peak torque during exercise were not correlated with changes in MVC, or other markers. CONCLUSION: These results suggest the large inter-subject variability in responses to eccentric exercise is not associated with work performed or torque generated during eccentric exercise.

An abdominal mass owing to Penicillium marneffei in an HIV-infected 7-year-old boy: case report.

A 7-year-old boy, referred with lymphoma, presented with prolonged fever and intra-abdominal lymphadenopathy demonstrated on computed tomography (CT) of the abdomen. Blood culture isolated Penicillium marneffei. The patient was subsequently proven serologically to be positive for human immunodeficiency virus (HIV). Treatment with amphotericin B followed by itraconazole was successful. A high level of clinical suspicion and awareness is necessary for early diagnosis of penicilliosis, especially in an era of an increasing prevalence of HIV in this region.

Malaysian Indians are genetically similar to Caucasians: CYP2C9 polymorphism.

BACKGROUND: CYP2C9 is one of the major drug metabolizing enzymes for many drugs including warfarin, NSAIDs and losartan. It is polymorphic in many populations. Data on the distribution of CYP2C9 and the implication of CYP2C9 polymorphism in the Malaysian population is lacking. Our objectives were therefore to investigate the prevalence of CYP2C9 variants among unrelated healthy volunteers of Malays, Chinese and Indians in Malaysia. METHOD: Deoxyribonucleic acid was extracted using standard lysis methods. Allele specific polymerase chain reaction was performed for determination of CYP2C9*1, *2, *3, *4 and *5 variants according to Z. Zainuddin, L.K. Teh, A.W.M. Suhaimi, M.Z. Salleh, R. Ismail (2003, Clinica Chimica Acta, 336, 97). RESULT: The Chinese had the highest frequency of CYP2C9*1 (321/330, 97.27%), followed by the Malays and the Indians (402 of 420, 95.71% and 291 of 330, 88.18%, respectively). CYP2C9*2 was not found in the Chinese. CYP2C9*3 were detected in all the three races with the Indians having the highest frequency of CYP2C9*3 (9.7%). The Indians had a frequency of CYP2C9*2 and *3 similar to Tamilians and Caucasians. Two of the Indians had *2/*3 and one had *3/*3 genotypes and are likely to be slow metabolizers. No subject with CYP2C9*4 and *5 were detected in our populations. CONCLUSION: CYP2C9*2 and *3 were identified in our population. Indians are similar to Caucasians in terms of CYP2C9 genotypes and thus may respond to CYP2C9 substrates differently when compared with the Malays and Chinese in Malaysia.

Effect of pulping variables on the characteristics of oil-palm frond-fiber.

Caustic pulping of oil-palm frond-fiber strands was conducted following a central composite design using a two-level factorial plan involving three pulping variables (temperature: 160-180 degrees C, time: 1-2 h, alkali charge: 20-30% NaOH). Responses of pulp properties to the process variables were analyzed using a statistical software (DESIGN-EXPERT). The results indicated that frond-fiber strands could be pulped with ease to about 35-45% yield. Statistically, the reaction time was not a significant factor while the influences of the treatment temperature and caustic charge were in general significantly relative to the properties of the resultant pulps.

A simple method for the detection of CYP2C9 polymorphisms: nested allele-specific multiplex polymerase chain reaction.

BACKGROUND: Cytochrome P4502C9 (CYP2C9), a principle drug-metabolizing enzyme is polymorphic in humans and is responsible for important pharmacokinetic and pharmacodynamic variations of CYP2C9 substrates. We developed an allele-specific multiplex polymerase chain reaction (PCR) method for the detection of common CYP2C9 alleles. METHOD: Genomic DNA was extracted from blood obtained from 40 unrelated healthy Malaysian Indian volunteers. The DNA was subjected to a first PCR that was used to amplify both exons 3 and 7 simultaneously in one reaction tube and a second PCR that was used to detect the polymorphic sites of CYP2C9 alleles using allele-specific primers. Sequencing was performed to validate the test results. RESULTS: We were successful in amplifying the fragments of interest from the DNA samples. The method was also reproducible and specific. The amplified sequences showed 100% homology to CYP2C9 sequence. CONCLUSION: This is the first nested allele-specific multiplex PCR method reported to allow for the simultaneously detection of five CYP2C9 alleles.

Evaluation of semen collected by electroejaculation from captive lesser Malay chevrotain (Tragulus javanicus).

Thirteen sexually mature captive male lesser Malay chevrotains (Tragulus javanicus) were each anesthetized twice with tiletamine-zolazepam for electroejaculation. Viable spermatozoa were collected from all animals. The semen was creamy, milky, pale yellowish, or watery. The mean values for ejaculate volume, sperm concentration, and percentages of sperm motility, normality and viability were 23.7 +/- 2.5 microl, 366.9 +/- 127.8 x 10(6) spermatozoa/ml, 40.0% +/- 3.1%, 71.4% +/- 1.6%, and 59.6% +/- 2.1%, respectively. Semen pH was 7-8. No adverse effects of electroejaculation were noted. These are the first reported values for semen of lesser Malay chevrotain. Electroejaculation should be usable for routine semen collection in this species.

Molecular epidemiology of tuberculosis in Malaysia.

Molecular typing with IS6110 was applied to Mycobacterium tuberculosis isolates from all parts of Malaysia. The degree of clustering increased with patient age, suggesting that reactivation may contribute to clustering. Identical banding patterns were also obtained for isolates from widely separate regions. Therefore, the use of clustering as a measure of recent transmission must be treated with caution. Strains related to the Beijing family were common in Peninsular Malaysia but were less common in Sabah and Sarawak, while a distinct group of strains comprised nearly 40% of isolates from East Malaysia but such strains were rare in Peninsular Malaysia. Single-copy strains, common in South and Southeastern Asia, constituted nearly 20% of isolates from the peninsula but were virtually absent in East Malaysia. The marked geographical difference in the prevailing strains indicates not only a restricted dissemination of M. tuberculosis but also a considerable degree of stability in the banding patterns.

Conservation of IS6110 sequence in strains of Mycobacterium tuberculosis with single and multiple copies.

The insertion sequence IS6110/IS986 is used extensively for detecting and typing Mycobacterium tuberculosis. In order to appreciate the evolutionary and epidemiological significance of differences in the fingerprint pattern, it is necessary to understand the factors influencing transposition frequency. Although most strains have many copies of this element, some have only one or two copies; it has been suggested that this apparent transpositional defect arises from the minor differences in the sequence of the element from different strains. In contrast, we have found that the sequence is identical in high and low copy number strains, indicating that external factors, such as the presence of adjacent promoters, must be responsible for differences in copy number.

Insertion sequence typing of Mycobacterium tuberculosis: characterization of a widespread subtype with a single copy of IS6110.

DNA fingerprinting with the insertion sequence IS6110 (also known as IS986) has become established as a major tool for investigating the spread of tuberculosis. Most strains of Mycobacterium tuberculosis have multiple copies of IS6110, but a small minority carry a single copy only. We have examined selected strains from Malaysia, Tanzania and Oman, in comparison with M. bovis isolates and BCG strains carrying one or two copies of IS6110. The insertion sequence appears to be present in the same position in all these strains, which suggests that in these organisms the element is defective in transposition and that the loss of transposability may have occurred at an early stage in the evolution of the M. tuberculosis complex.

Characterization of a Mycobacterium tuberculosis insertion sequence belonging to the IS3 family.

A repetitive element (IS986), previously isolated from Mycobacterium tuberculosis and shown to detect multiple restriction fragment-length polymorphisms (RFLPs), has been sequenced. It consists of a potential insertion sequence of 1358bp, with 30-bp inverted repeat ends. IS986 has four potentially significant open reading frames (ORFs): ORFa1, ORFa2 and ORFb on one strand and ORFc on the complementary strand. The sequences of the potential translated products identify IS986 as a member of the IS3 family, with an apparent frameshift between ORFa1 and ORFa2. IS986 has potential as a highly specific probe for detection and typing of M. tuberculosis, as well as for transposon mutagenesis of mycobacteria. The sequence of IS986 is virtually identical to that of another recently described element, IS6110 (Thierry et al., 1990).

Does Mycobacterium tuberculosis have plasmids?

Evidence of the presence of plasmids in Mycobacterium tuberculosis is lacking, whereas they are widespread in some other mycobacterial species. We examined, by agarose gel electrophoresis, a total of 197 clinical isolates of M. tuberculosis, mostly resistant to one or more antibiotics, and were able to detect bands of apparently extrachromosomal DNA at a low level in some isolates. These presumptive plasmids could not be isolated by CsCl/ethidium bromide gradient ultracentrifugation, and may consist of unusual forms of DNA. The possible existence of single stranded plasmid DNA is discussed.

Polymorphic repetitive DNA sequences in Mycobacterium tuberculosis detected with a gene probe from a Mycobacterium fortuitum plasmid.

Clinical isolates of Mycobacterium tuberculosis were shown by Southern blotting to contain DNA sequences hybridizing to a probe derived from a Mycobacterium fortuitum plasmid. Two such M. tuberculosis DNA fragments, isolated from a gene library, were used as probes to show restriction fragment length polymorphism in M. tuberculosis strains by detecting a repetitive sequence apparently located at different points on the chromosome. This could indicate the presence of a transposable element in M. tuberculosis which is partly homologous to a region of the M. fortuitum plasmid. The probes described can be used to fingerprint M. tuberculosis isolates, and in addition are capable of distinguishing M. tuberculosis from Mycobacterium bovis and BCG.

Transformation of Mycobacterium smegmatis with E. coli plasmids.

One limiting factor in the studies of tuberculosis and leprosy is the lack of a versatile system for genetic analysis and manipulation of mycobacteria. One strategy used in constructing a plasmid vector for transforming Mycobacterium smegmatis was to insert fragments of a mycobacterial plasmid into an Escherichia coli plasmid. We found that the parental E. coli plasmid is capable of self-replication in M. smegmatis yielding chloramphenicol-resistant colonies. Plasmids from different passages of one M. smegmatis transformant were recovered and characterised by restriction digest analysis. The plasmid from the earlier passage was found to be indistinguishable from the original plasmid by restriction analysis. Plasmids from later preparations, however, were found to have undergone modifications in the M. smegmatis host resulting in an apparent increase in transformation efficiency for M. smegmatis. These plasmids can be used as a shuttle vector for the genetic manipulation of mycobacterial species.

Transformation of Mycobacterium smegmatis with Escherichia coli plasmids carrying a selectable resistance marker.

One limiting factor in studies of tuberculosis and leprosy is the difficulty of genetic analysis and manipulation of mycobacteria. Two approaches were adopted for the construction of vectors, based on different Escherichia coli plasmids and using Mycobacterium smegmatis as the host. In both cases we found that the original E. coli plasmid is capable of being replicated in M. smegmatis, yielding chloramphenicol-resistant colonies. One such plasmid has been recovered from a M. smegmatis transformant and used to re-transform both M. smegmatis and E. coli to chloramphenicol resistance. This plasmid is indistinguishable from the original plasmid by restriction analysis, and can be used as a shuttle vector for the genetic manipulation of mycobacterial species.