Five out of the nine benzene-toulene-ethylbenzene-xylene (BTEX) tolerant bacteria that demonstrated high protease activity on skim milk agar were isolated. Among them, isolate 115b identified as Bacillus pumilus exhibited the highest protease production. The protease produced was stable in 25% (v/v) benzene and toluene and it was activated 1.7 and 2.5- fold by n-dodecane and n-tetradecane, respectively. The gene encoding the organic solvent tolerant protease was cloned and its nucleotide sequence determined. Sequence analysis revealed an open reading frame (ORF) of 1,149 bp that encoded a polypeptide of 383 amino acid residues. The polypeptide composed of 29 residues of signal peptide, a propeptide of 79 residues and a mature protein of 275 amino acids with a calculated molecular mass of 27,846 Da. This is the only report available to date on organic solvent tolerant protease from B. pumilus.
Superior mesenteric artery syndrome (SMAS) is a rare complication following correction of scoliosis with either nonoperative or operative methods. If the patient diagnosed with this syndrome is not managed timely and adequately, mortality may result. We report two cases of SMAS complicating staged corrective surgery for scoliosis using modern segmental derotation instrumentation system. The aim of this report is to highlight the clinical presentations, laboratory findings, radiologic features, and management of the syndrome. The first patient had the syndrome after two-staged scoliosis surgery with halo traction between two stages, and the second patient after three-staged scoliosis surgery with halo traction between the first and second surgeries. The first patient responded well to conservative treatment. However, the second patient failed to respond to conservative treatment and needed a gastrojejunostomy operation to bypass the duodenal obstruction. Clinicians treating post scoliosis surgery patients should always have a high index of suspicion for this potential life-threatening condition. Early diagnosis will enable a multidisciplinary team approach to be initiated early to provide optimal care for the patient. Nutritional and fluid supplementation is mandatory during conservative treatment. The duration for trial of conservative treatment should not exceed 1 week.
Ice cream mixes containing 33.4% total solids including 10% fat, 11.1% milk solid-non fat (MSNF), 12% sugar, 0.35% commercial blend of emulsifier/ stabiliser and water were produced. The blending of PO with AMF were conducted at three different ratios 30: 70, 50: 50 and 70: 30, respectively. The experimental ice cream mixes were compared with a control ice cream mix prepared from AMF. The flow properties were measured after ageing at 0, 1, 1.5, 2 and 24 h and determined using a controlled stress rheometer (Haake RS 100). The Power Law and Casson equation was employed to estimate the yield stress of an ice cream mixes. The regression coefficients (r) was represented well by the Casson model (r > 0.99) for all the samples, indicating goodness of fit. The profiles of the consistency coefficients (K(c)) were quite similar for all experimental samples, which could be attributed to the fact that all the samples exhibited similar viscoelastic behaviour. The flow behaviour index (n) of an ice cream mix prepared from PO and their blends with AMF were less then 1.0 (range 0.04-0.08) indicating that they were psuedoplastic fluid. The eta(o) at shear rate 20(-1) indicated higher degree of viscosity in AMF.
Between December 2003 and January 2004 highly pathogenic avian influenza (HPAI) H5N1 infections of poultry were declared in China, Japan, South Korea, Laos, Thailand, Cambodia, Vietnam, and Indonesia. In 2004 an outbreak was reported in Malaysia. In 2005 H5N1 outbreaks were recorded in poultry in Russia, Kazakhstan, Mongolia, Romania, Turkey, and Ukraine, and virus was isolated from swans in Croatia. In 2004 HPAI H5N1 virus was isolated from smuggled eagles detected at the Brussels Airport and in 2005 imported caged birds held in quarantine in England. In 2006 HPAI was reported in poultry in Iraq, India, Azerbaijan, Pakistan, Myanmar, Afghanistan, and Israel in Asia; Albania, France, and Sweden in Europe; and Nigeria, Cameroon, and Niger in Africa; as well as in wild birds in some 24 countries across Asia and Europe. In 2003, over 25,000,000 birds were slaughtered because of 241 outbreaks of HPAI caused by virus of H7N7 subtype in the Netherlands. The virus spread into Belgium (eight outbreaks) and Germany (one outbreak). HPAI H5N2 virus was responsible for outbreaks in ostriches in South Africa during 2005. HPAI H7N3 virus was isolated in Pakistan in 2004. Low-pathogenicity avian influenza (LPAI) H5 or H7 viruses were isolated from poultry in Italy (H7N3 2002-2003; H5N2 2005), The Netherlands (H7N3 2002), France (H5N2 2003), Denmark (H5N7 2003), Taiwan (H5N2 2004), and Japan (H5N2 2005). Many isolations of LPAI viruses of other subtypes were reported from domestic and wild birds. Infections with H9N2 subtype viruses have been widespread across Asia during 2002-06.
MeSH terms: Africa/epidemiology; Animals; Asia/epidemiology; Birds/virology; Disease Outbreaks/veterinary; Europe/epidemiology; Influenza in Birds/epidemiology*; Influenza in Birds/virology*; Time Factors; Australasia; Influenza A Virus, H5N1 Subtype/classification; Influenza A Virus, H5N1 Subtype/isolation & purification*; Influenza A Virus, H5N1 Subtype/pathogenicity
Numerous lessons have been learned so far in controlling H5N1 avian influenza in Asia. Early detection of incursions of virus prevented establishment of the disease in several countries, notably Japan, South Korea, and Malaysia. In countries where detection of early cases was delayed, infection is endemic and has been for three or more years. Control measures implemented in these countries need to reflect this finding. Vaccination will continue to be one of the key measures used in these endemically infected countries. Used alone, vaccination will not result in elimination of H5N1 viruses from a country, but, if used correctly, it will markedly reduce the prevalence of and susceptibility to infection. Vaccination has already played a valuable role in reducing the adverse effects of H5N1 viruses. Mass culling also reduces the level of infection in infected areas. However, the long-term benefits are limited in endemically infected countries owing to the high probability of reinfection on restocking unless other measures are used in parallel. Full epidemiological studies have not been conducted in many infected countries. Nevertheless, it is recognized that the number of clinical cases does not truly reflect the levels of infection. Domestic ducks and large live poultry markets have played a key role in the persistence of infection, because they can be infected silently. In tackling this disease, countries should adopt integrated control programs using the combination of measures best suited to the local environment. All surveillance data should be shared, both positive and negative, and should include information on cases of infection and disease. Socioeconomic and ecological implications of all control measures should be assessed before implementation, especially the impact on the rural poor.
MeSH terms: Agriculture; Animals; Asia/epidemiology; Birds; Communicable Disease Control/methods*; Disease Outbreaks/veterinary*; Environment; Influenza in Birds/epidemiology*; Influenza in Birds/prevention & control; Influenza in Birds/virology*; Influenza Vaccines/immunology; Population Surveillance; Socioeconomic Factors; Influenza A Virus, H5N1 Subtype/isolation & purification; Influenza A Virus, H5N1 Subtype/physiology*
A fast and simple capillary zone electrophoresis method was developed and validated for the determination of lidocaine in skin using tape samples. Separation was performed in a 350 mm (265 mm to window) x 50 microm i.d. fused silica capillary using a background electrolyte of phosphoric acid-Tris pH 2.5. The extraction of lidocaine from tape samples was achieved using methanol, which was diluted to 50% with water before injection. Procaine was the internal standard. The migration times for procaine and lidocaine were 2.9 and 3.2 min, respectively. The limit of quantification for lidocaine was 50 microg, with signal to noise ratio greater than 10. The calibration curve was linear from 50 to 1000 microg with r(2) greater than 0.99. The CV for both within- and between-assay imprecision and the percentage of inaccuracy for the quality control samples including lower and upper limits of quantitation were 97%. The accuracy and selectivity of this method allowed the measurement of lidocaine in tape samples obtained from a skin tape stripping study of local anesthetics in healthy subjects.
MeSH terms: Anesthetics, Local/analysis*; Calibration; Humans; Lidocaine/analysis*; Sensitivity and Specificity; Skin/chemistry*; Specimen Handling/methods*; Reproducibility of Results; Electrophoresis, Capillary/methods*
The impacts of global change on tropical forests remain poorly understood. We examined changes in tree growth rates over the past two decades for all species occurring in large (50-ha) forest dynamics plots in Panama and Malaysia. Stem growth rates declined significantly at both forests regardless of initial size or organizational level (species, community or stand). Decreasing growth rates were widespread, occurring in 24-71% of species at Barro Colorado Island, Panama (BCI) and in 58-95% of species at Pasoh, Malaysia (depending on the sizes of stems included). Changes in growth were not consistently associated with initial growth rate, adult stature, or wood density. Changes in growth were significantly associated with regional climate changes: at both sites growth was negatively correlated with annual mean daily minimum temperatures, and at BCI growth was positively correlated with annual precipitation and number of rainfree days (a measure of relative insolation). While the underlying cause(s) of decelerating growth is still unresolved, these patterns strongly contradict the hypothesized pantropical increase in tree growth rates caused by carbon fertilization. Decelerating tree growth will have important economic and environmental implications.
We investigate the geographical and historical context of diversification in a complex of mutualistic Crematogaster ants living in Macaranga trees in the equatorial rain forests of Southeast Asia. Using mitochondrial DNA from 433 ant colonies collected from 32 locations spanning Borneo, Malaya and Sumatra, we infer branching relationships, patterns of genetic diversity and population history. We reconstruct a time frame for the ants' diversification and demographic expansions, and identify areas that might have been refugia or centres of diversification. Seventeen operational lineages are identified, most of which can be distinguished by host preference and geographical range. The ants first diversified 16-20 Ma, not long after the onset of the everwet forests in Sundaland, and achieved most of their taxonomic diversity during the Pliocene. Pleistocene demographic expansions are inferred for several of the younger lineages. Phylogenetic relationships suggest a Bornean cradle and major axis of diversification. Taxonomic diversity tends to be associated with mountain ranges; in Borneo, it is greatest in the Crocker Range of Sabah and concentrated also in other parts of the northern northwest coast. Within-lineage genetic diversity in Malaya and Sumatra tends to also coincide with mountain ranges. A series of disjunct and restricted distributions spanning northern northwest Borneo and the major mountain ranges of Malaya and Sumatra, seen in three pairs of sister lineages, further suggests that these regions were rain-forest refuges during drier climatic phases of the Pleistocene. Results are discussed in the context of the history of Sundaland's rain forests.
Once a very slowly developing country in a Southeast Asia region, Malaysia has undergone considerable change over the last 20 years after the government changed its focus from agriculture to developing more industry and technology. The well-known "Vision 2020," introduced by the late Prime Minister, set a target for the nation to be a developed country in the Asia region by the year 2020. As the economy and standard of living have improved, the demand from the public for a better health care system, in particular, emergency medical services (EMS), has increased. Despite the effort by the government to improve the health care system in Malaysia, EMS within the country are currently limited, best described as being in the "developing" phase. The Ministry of Health, Ministry of Education, Civil Defense, and non-governmental organizations such as Red Crescent and St. John's Ambulance, provide the current ambulance services. At the present time, there are no uniform medical control or treatment protocols, communication systems, system management, training or education, or quality assurance policies. However, the recent development of and interest in an Emergency Medicine training program has gradually led to improved EMS and prehospital care.
MeSH terms: Ambulances/organization & administration; Ambulances/supply & distribution*; Delivery of Health Care; Developing Countries; Emergency Medical Services/organization & administration*; Emergency Medical Services/standards; Emergency Medical Services/supply & distribution; Emergency Medical Technicians/education*; Emergency Medical Technicians/supply & distribution; Emergency Medicine/education; Humans; Malaysia; Quality Assurance, Health Care
The expression of high-abundance serum proteins in newly diagnosed patients with endometrial adenocarcinoma (EACa), squamous cell cervical carcinoma (SCCa) and cervical adenocarcinoma (ACCa), relative to control female subjects, was analyzed by subjecting serum samples to 2-DE followed by image analysis of the silver-stained protein profiles. The three cohorts of cancer patients demonstrated different altered expression of serum high-abundance proteins compared to negative control women. The expression of alpha1-antitrypsin, alpha1-B glycoprotein, cleaved high-molecular-weight kininogen (light chain) and antithrombin III were consistently altered in all the patients. However, clusterin was upregulated only in the patients with EACa, while those with SCCa and ACCa were typically characterized by the upregulated expression of zinc alpha-2-glycoprotein. The aberrant expression of selective serum proteins in the various cohorts of cancer patients was validated by competitive ELISA as well as by lectin detection. Analysis by using the champedak galactose binding lectin further highlighted an unidentified protein that may be differently glycosylated in the sera of the EACa patients that were studied.
The UGT1A1 Taqman MGB probe single nucleotide polymorphism (SNP) genotyping assay was developed to detect nucleotide 211 of the UDP-glucoronocyltransferase 1A1 (UGT1A1) gene. Defects in this enzyme interfere with process of conjugation of bilirubin and cause unconjugated hyperbilirubinemia. Variation at nucleotide 211 in the coding region of the UGT1A1 gene has been shown to be prevalent in Japanese and Chinese. Using an ABI sequence detection system (SDS) 7000, an allele-specific real-time PCR-based genotyping method was established to detect nucleotide G211A. Cord blood from 125 infants without hyperbilirubinemia (controls) were compared with cord blood from 74 infants (cases) with severe hyperbilirubinemia (total serum bilirubin > 300 micromol/L). Homozygous variation of the UGT1A1 gene at nucleotide 211(A/A) is significantly more common in cases (14.9%) than in controls (0.8%) (P<0.001). Direct sequencing from 20 randomly selected samples showed eight samples with homozygous wild type, seven with homozygous variant, and five samples were heterozygous. The result from this assay was in complete concordance with the DNA sequencing result and clearly discriminate wild-type (G/G), homozygous variant (A/A), and heterozygous (G/A). This assay is rapid and robust for screening of SNP G211A to determine if this polymorphism plays a role in causing severe neonatal jaundice in the local context.
Brain tumorigenesis is a complex process involving multiple genetic alterations. Cyclin D1 and BAX genes are two of the most important regulators in controlling the normal proliferation and apoptosis of cells, respectively. In this study, we analysed the possibilities of involvement of cyclin D1 and BAX genes in the gliomagenesis.
MeSH terms: DNA Mutational Analysis/methods; Exons; Glioma/genetics*; Humans; Mutation; Retrospective Studies; RNA, Messenger/metabolism; Central Nervous System Neoplasms/genetics*; Cyclin D1/genetics*; Reverse Transcriptase Polymerase Chain Reaction; bcl-2-Associated X Protein/genetics*
Measurements of external radiation level, radon/thoron daughters concentrations in air and uranium/thorium concentrations in airborne mineral dust at 16 amang plants in Malaysia were carried out for three consecutive months to assess radiation dose to workers. Estimated occupational dose was within the range of 1.7-10.9 mSv y(-1). The mean total dose at the amang plants was 4.1 mSv y(-1). Overall, it was found that the major dose contribution of 80% came from external radiation. Radon/thoron daughters and airborne mineral dust contributed to only 11 and 9% of the total dose, respectively.
Delayed diagnosis of tuberculosis (TB) can lead to an increased period of infectivity in the community, a delay in treatment and a severe form of the disease. The objective of this study was to determine the length of delay, and factors linking the delay from the onset of symptoms of pulmonary tuberculosis (PTB) until the commencement of treatment in Sarawak, Malaysia.
MeSH terms: Adolescent; Adult; Aged; Cross-Sectional Studies; Ethnic Groups/statistics & numerical data; Female; Humans; Malaysia/epidemiology; Male; Middle Aged; Rural Population/statistics & numerical data; Self Care/utilization; Severity of Illness Index; Time Factors; Tuberculosis, Pulmonary/diagnosis*; Tuberculosis, Pulmonary/epidemiology*; Tuberculosis, Pulmonary/therapy; Logistic Models; Sex Distribution; Health Care Surveys; Continental Population Groups/statistics & numerical data
Avian influenza viruses are pathogens of economical and public health concerns. However, infections caused by low pathogenic avian influenza particularly H9N2 subtype are not associated with clear clinical features. Hence, rapid detection and subtyping of the virus will enable immediate measures to be implemented for preventing widespread transmission. This study highlights the development of a multiplex real-time reverse-transcriptase polymerase chain reaction (RRT-PCR) assay using SYBR Green 1 chemistry for universal detection of avian influenza viruses and specific subtyping of H9N2 isolates based on melting temperatures (T(m)) discriminations. Three melting peaks generated simultaneously at temperatures 85.2+/-1.0, 81.9+/-0.9 and 78.7+/-0.9 degrees C represent NP, H9 and N2 gene products, respectively. The RRT-PCR assay was about 10-100-fold more sensitive when compared to the conventional RT-PCR method using reference H9N2 isolate. In addition, the RRT-PCR assay was 100% sensitive as well as 92% specific according to the standard virus isolation method in detecting experimentally infected specific-pathogen-free (SPF) chickens.
MeSH terms: Animals; Chickens*; Influenza in Birds/diagnosis*; Organic Chemicals; RNA, Viral/analysis; Sensitivity and Specificity; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction/methods*; Influenza A Virus, H9N2 Subtype/classification; Influenza A Virus, H9N2 Subtype/genetics; Influenza A Virus, H9N2 Subtype/isolation & purification*
The present work examined the effect of chronic oral administration of quercetin, a flavonoid antioxidant, on blood glucose, vascular function and oxidative stress in STZ-induced diabetic rats. Male Wistar-Kyoto (WKY) rats were randomized into euglycemic, untreated diabetic, vehicle (1% w/v methylcellulose)-treated diabetic, which served as control, or quercetin (10mgkg(-1) body weight)-treated diabetic groups and treated orally for 6 weeks. Quercetin treatment reduced blood glucose level in diabetic rats. Impaired relaxations to endothelium-dependent vasodilator acetylcholine (ACh) and enhanced vasoconstriction responses to alpha(1)-adrenoceptor agonist phenylephrine (PE) in diabetic rat aortic rings were restored to euglycemic levels by quercetin treatment. Pretreatment with N(omega)-nitro-l-arginine methyl ester (l-NAME, 10microM) or methylene blue (10microM) completely blocked but indomethacin (10microM) did not affect relaxations to ACh in aortic rings from vehicle- or quercetin-treated diabetic rats. PE-induced vasoconstriction with an essentially similar magnitude in vehicle- or quercetin-treated diabetic rat aortic rings pretreated with l-NAME (10microM) plus indomethacin (10microM). Quercetin treatment reduced plasma malonaldehyde (MDA) plus 4-hydroxyalkenals (4-HNE) content as well as increased superoxide dismutase activity and total antioxidant capacity in diabetic rats. From the present study, it can be concluded that quercetin administration to diabetic rats restores vascular function, probably through enhancement in the bioavailability of endothelium-derived nitric oxide coupled to reduced blood glucose level and oxidative stress.
To evaluate the perioperative complications associated with surgical correction in neuromuscular scoliosis and to identify the risk factors associated with these complications.
The first stage in Peyer's patch development in the fetal lamb is characterized by the colonization of the rudimentary Peyer's patches by precursor cells expressing the IgM surface receptor. In the fetal lamb, the spleen has been implicated as the source of gene-rearranged IgM(+) B lymphocytes. This study was intended to quantitate IgM(+) lymphocytes in the spleen, lymph nodes and liver of fetal lambs at various gestational ages between 63 and 110 days using flow cytometry. Flow-cytometric analysis revealed that IgM(+) lymphocytes were rare in the liver being consistently less than 1% at every gestational age examined. IgM(+) lymphocytes were detected in the spleen (mean 9.18%) and prescapular lymph nodes (mean 11.89%) as early as 63 days. In both spleen and lymph nodes, the highest representation of IgM(+) lymphocytes occurred between 70 and 86 days gestation. The highest mean percentage of IgM(+) lymphocytes was observed in the spleen (22.63%) and lymph nodes (17.02%) at 75 days gestation. From 98 days onwards, B-lymphocyte density gradually decreased in both spleen and prescapular lymph nodes. This study indicates that substantial populations of IgM(+) lymphocytes were present in both the spleen and prescapular lymph nodes from 70 days gestation and implies that both of these locations could be potential sources for the normal colonization of the ileal Peyer's patches.