Male-specific RNA coliphages (FRNA) have been recommended as indicators of fecal contamination and of the virological quality of water. In this study, 16 river water and 183 animal fecal samples were examined for the presence of FRNA coliphages by a plaque assay using Salmonella typhimurium WG49 and WG25 to differentiate between male-specific and somatic phages, a RNase spot test to differentiate between DNA and RNA phages and a reverse transcriptase-polymerase chain reaction (RT-PCR) for the specific identification of FRNA phages. The overall recovery rate for F-specific coliphages was 8.0%. (4.4% from animal fecal matter and 50% from river water samples). Plaque counts were generally low (< 6 x 10(2) pfu per g feces or ml water), with FRNA (6.5%) and Male-specific DNA coliphages (FDNA) (7.0%) phages occurring at almost equal frequencies. The RT-PCR was positive in all FRNA plaques and was able to identify FRNA phages in mixed populations of FRNA, FDNA and somatic phages.
Many plant-derived natural compounds have been reported previously to inhibit the production of important pro-inflammatory mediators such as nitric oxide, prostaglandin E2, TNF-alpha and reactive oxygen species by suppressing inducible enzyme expression via inhibition of the mitogen-activated protein kinase pathway and nuclear translocation of critical transcription factors. This study evaluates the effects of atrovirinone [2-(1-methoxycarbonyl-4,6-dihydroxyphenoxy)-3-methoxy-5,6-di-(3-methyl-2-butenyl)-1,4-benzoquinone)], a benzoquinone that we have previously isolated from Garcinia atroviridis, on two cellular systems that are repeatedly used in the analysis of anti-inflammatory bioactive compounds, namely, RAW 264.7 macrophage cells and whole blood. Atrovirinone inhibited the production of both nitric oxide and prostaglandin E2 from LPS-induced and IFN-gamma-induced RAW 264.7 cells and whole blood, with inhibitory concentration (IC)50 values of 4.62 +/- 0.65 and 9.33 +/- 1.47 micromol/L, respectively. Analysis of thromboxane B2 (TXB2) secretion from whole blood stimulated by either the cyclooxygenase (COX)-1 or the COX-2 pathway showed that atrovirinone inhibits the generation of TXB2 by both pathways, with IC50 values of 7.41 +/- 0.92 and 2.10 +/- 0.48 micromol/L, respectively. Analysis of IC50 ratios showed that atrovirinone was more COX-2 selective in its inhibition of TXB2, with a ratio of 0.32. Atrovirinone also inhibited the generation of intracellular reactive oxygen species and the secretion of TNF-alpha from RAW 264.7 cells in a dose-responsive manner, with IC50 values of 5.99 +/- 0.62 and 11.56 +/- 0.04 micromol/L, respectively. Lipoxygenase activity was also moderately inhibited by atrovirinone. Our results suggest that atrovirinone acts on important pro-inflammatory mediators possibly by the inhibition of the nuclear factor-kappaB pathway and also by the inhibition of the COX/lipoxygenase enzyme activity.
Human Immunodeficiency Virus (HIV)-related pulmonary hypertension is a relatively rare disease that can affect HIV sufferers. This is almost always associated with a poor outcome and death. An 18 month-old girl, probably the youngest on record, was diagnosed to have pulmonary hypertension (PHT) and retrospectively found to have HIV infection. Sildenafil was used to control her PHT and she remains alive even after 2 years.
Tuberculosis remains a major infectious disease with over 8 million new cases and 2 million deaths annually. Therefore, a vaccine more potent than BCG is desperately needed. In this regard, an approximately 800 bp DNA encoding a mycobacterial synthetic gene designated as VacIII (containing ubiquitin gene UbGR and four immunogenic mycobacterial epitopes or genes of ESAT-6, Phos1, Hsp 16.3, and Mtb8.4) was sub-cloned into a bacterial expression vector of pRSET-B resulting in a 6 x His-VacIII fusion gene construction. This recombinant clone was over expressed in Escherichia coli BL-21 (DE-3). The expressed fusion protein was found almost entirely in the insoluble form (inclusion bodies) in cell lysate. The inclusion bodies were solubilized with 8M urea and the recombinant protein was purified by Ni-NTA column and dialyzed by urea gradient dialysis. This method produced a relatively high yield of recombinant VacIII protein and the cloned VacIII gene offers the potential development of other vaccine formats such as DNA vaccine and recombinant vaccine.
Continuous outbreaks of the highly pathogenic H5N1 avian influenza A in Asia has resulted in an urgent effort to improve current diagnostics to aid containment of the virus and lower the threat of a influenza pandemic. We report here the development of a PCR-based assay that is highly specific for the H5N1 avian influenza A virus.
MeSH terms: Cross Reactions; Humans; Sensitivity and Specificity; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction/methods*; Influenza A Virus, H5N1 Subtype/genetics*; Influenza A Virus, H5N1 Subtype/isolation & purification*
Chinese and Malay subjects have been reported to require less maintenance warfarin than Indians that could not be accounted for by cytochrome P450 (CYP) 2C9 variants. Vitamin K epoxide reductase complex 1 (VKORC1) is the target enzyme of warfarin, and VKORC1 intronic variants and haplotypes have recently been shown to influence VKORC1 activity and warfarin requirements.
Computed tomography (CT) perfusion is a new method to diagnose ischaemic stroke especially in developing countries. It identifies the area and is useful to predict the size of final infarction. The aim of this study was to assess cerebral ischaemia with CT perfusion (CTP) among patients with acute ischaemic stroke in Hospital Universiti Sains Malaysia, a tertiary referral centre in a rural setting.
A 15-year-old girl, who was previously well, complained of a mass in the abdomen after a minor motor vehicle accident. Physical and radiological investigations revealed a mass in the body of pancreas containing proteinaceous material and multiple nodules in both lobes of liver. Serological investigations for malignancy were normal. Histopathological examination of the resected specimen showed pancreatoblastoma. Pancreatoblastoma is an unusual malignant tumour seen in infants and children although rare cases have also been reported in adults. They are clinicopathologically distinct from adult pancreatic ductal carcinoma. The histogenesis, clinical features and treatment options are discussed along with presentation of the case.
Shell morphological characters and allozyme electrophoresis were used to study the relationships among six geographical populations of land snails collected from Peninsular Malaysia. Allozyme electrophoresis was used to study the genetic variations to complement the morphological features studied that included shell lengths, numbers of whorls and shell colour. Ten loci coding for six enzymes (MDH, LAP, ALP, PGM, G6PDH and EST) could be reliably scored in samples from the six populations studied. The dendrogram showed two major clusters with one cluster comprising Subulinidae populations from Perak, Selangor, Johor, Terengganu and Pahang while the other cluster included only the Streptaxidae Huttonella bicolor (red) population. The Subulinidae populations were grouped into two subclusters: one subcluster included the Subulina sp. populations from Perak, Selangor an Johor while the other subcluster included the Opeas sp. populations from Terengganu and Pahang. Morphological features can identify the different families and therefore they can complement the allozyme genetic studies on the land snail populations. Like other reports in the literature, our results also underline the importance of a genetic approach in conjunction with a morphological approach, for discriminating land snail species. The present results suggest that small land snails, which were similar in colour but different in sizes, were not of the same family/genus.
Studies were carried out on heat treatment of bamboo species Gigantochloa scortechinii Gamble using palm oil. The samples were laminated and glued. The adhesion results showed that the delamination of glue line increased as the temperature and duration of oil heat treatment increased. Maximum load and shear strength of the glue line reduced as the heat treatment become more severe. It was found that the palm oil used as the heating medium penetrated in some parts of the cell wall as well as in the cell lumen of the bamboo.
Comparative molecular dynamics simulations of n-octyl-beta-D-galactopyranoside (beta-C8Gal) and n-octyl-beta-D-glucopyranoside (beta-C8Glc) micelles in aqueous solution have been performed to explore the influence of carbohydrate stereochemistry on glycolipid properties at the atomic level. In particular, we explore the hypothesis that differences in T(m) and T(c) for beta-C8Gal and beta-C8Glc in lyotropic systems arise from a more extensive hydrogen bonding network between beta-C8Gal headgroups relative to beta-C8Glc, due to the axial 4-OH group in beta-C8Gal. Good agreement of the 13 ns micelle-water simulations with available experimental information is found. The micelles exhibit a similar shape, size, and degree of exposed alkyl chain surface area. We find net inter- and intra-headgroup hydrogen bonding is also similar for beta-C8Gal and beta-C8Glc, although n-octyl-beta-D-galactopyranoside micelles do exhibit a slightly greater degree of inter- and intra-headgroup hydrogen bonding. However, the main distinction in the calculated microscopic behavior of beta-C8Glc and beta-C8Gal micelles lies in solvent interactions, where beta-d-glucosyl headgroups are considerably more solvated (mainly at the equatorial O4 oxygen). These results agree with preceding theoretical and experimental studies of monosaccharides in aqueous solution. A number of long water residence times are found for solvent surrounding both micelle types, the largest of which are associated with surface protrusions involving headgroup clusters. Our simulations, therefore, predict differences in hydrogen bonding for the two headgroup stereochemistries, including a small difference in inter-headgroup interactions, which may contribute to the higher T(m) and T(c) values of beta-C8Gal surfactants relative to beta-C8Glc in lyotropic systems.
Chemical modification of natural rubber (NR) has frequently been attempted to improve the performance in specific application. 30% poly(methyl metacrylate) (PMMA) grafted into NR (MG30) has been explored as a potential candidate for polymer electrolytes. The complexation effect of salt and plasticizer in polymer host electrolytes had been investigated using FTIR. The carbonyl stretch of MG30 locates at 1729 cm-1, with the addition of lithium trimethanesulfonate (LiCF3SO3) salt, new band evolves at lower frequency region at 1643-1645 cm-1. The nondegenerate vibrational mode of nus(SO3) of salted electrolytes appearing at 1031-1034 cm-1 comes from 'free' trimethanesulfonate anions and the 1040-1046 cm-1 absorption from the monodentate ion paired with triflates. These indicate MG30-salt interaction. When MG30 and ethylene carbonate (EC) formed film, the CH3 asymmetric bend of MG30 appearing at 1447cm-1 is shifted to 1449 cm-1 in the EC-polymer complex. The CO stretching at 1729 cm-1 also shifted to 1728 cm-1. Hence, the EC-MG30 system is complexed to each other. EC-LiCF3SO3 interactions are indicated by the shifting of CO bending band of EC from 718 cm-1 being shifted to 720 cm-1 in the complex. In Li+-EC interaction where the ring breathing region at 897 cm-1 in EC has shifted to 899 cm-1 in EC-salt spectrum. The band appearing at 1643-1645 cm-1 due to the coordination of Li+
The incidence of candidaemia among immunocompromised patients in Malaysia is increasing at an alarming rate. Isolation of clinical strains that are resistant to fluconazole has also risen markedly. We report here the repeated isolation of Candida tropicalis from the blood of a neonatal patient with Hirschsprung's disease. In vitro fluconazole susceptibility tests of the eight isolates obtained at different time points showed that seven of the isolates were resistant and one isolate was scored as susceptible dose-dependent. Random amplification of polymorphic DNA fingerprinting of the isolates using three primers and subsequent phylogenetic analysis revealed that these isolates were highly similar strains having minor genetic divergence, with a mean pairwise similarity coefficient of 0.893+/-0.041. The source of the infectious agent was thought to be the central venous catheter, as culture of its tip produced fluconazole-resistant C. tropicalis. This study demonstrates the utility of applying molecular epidemiology techniques to complement traditional mycological culture and drug susceptibility tests for accurate and appropriate management of recurrent candidaemia and highlights the need for newer antifungals that can combat the emergence of fluconazole-resistant C. tropicalis strains.
The proteomics approach was adopted to study the simultaneous expression of serum proteins in patients with nasopharyngeal carcinoma (NPC). We have subjected unfractionated whole sera of ten newly diagnosed Malaysian Chinese patients with WHO type III NPC to two-dimensional gel electrophoresis (2-DE) and image analysis. The results obtained were then compared to that generated from sera of ten normal healthy controls of the same ethnic group and range of age. Our data demonstrated that the serum high abundance 2-DE protein profiles of NPC patients were generally similar to that of the controls, with exception of the ceruloplasmin (CPL) spots (identified by mass spectrometric analysis and MASCOT database search), which showed higher expression. The enhanced expression of CPL in the patients' sera was confirmed by competitive ELISA. Immunohistochemical analysis of nasopharyngeal lesions of NPC patients demonstrated moderate to strong positive CPL staining in the cytoplasm of cells at the regions of malignancy but only weak cytoplasmic staining at normal epithelial lining areas. When follow-up 2-DE and ELISA studies were performed on five of the NPC patients who responded positively to six months treatment, the difference in CPL expression was no longer significant.
A Web-based service was set up to link subspecialist ophthalmologists with those seeking advice on particular clinical problems in ophthalmology. The service operated between countries, with centres in Malaysia, Ghana, Tanzania, South Africa and Gambia seeking advice from Moorfields Eye Hospital in the UK. It also operated within country, where a rural clinic in The Gambia sought advice from the central hospital on difficult cases or cases for possible referral. Provision of Web access and training in image capture and manipulation were undertaken in each participating centre. During the first 12 months, 132 cases were posted to the Website from five of the six centres participating. The rate of case referral rose to about 12-14 cases per month by the end of the study. Overall, 24% of referrals did not use images. In the first four months the response time was 13 days, and in the last four months it was three days. Most cases were answered with a single response from the specialist. The main problem was the amount of Internet down-time in each of the locations. The main benefit was safe and reliable access to specialist advice for practitioners.
MeSH terms: England; Eye Diseases/diagnosis*; Eye Diseases/therapy*; Gambia; Ghana; Humans; Interprofessional Relations; Malaysia; Medical Audit; Ophthalmology/methods*; Ophthalmology/statistics & numerical data; South Africa; Tanzania; Telemedicine/methods*; Telemedicine/utilization; Internet
Ant-garden (AG) associations are systems of epiphytic plants and arboricolous (i.e., tree-living) ants, in which the ants build fragile carton nests containing organic material. They collect and incorporate seeds or fruits of epiphytes that then germinate and grow on the nest [sensu Corbara et al. (1999) 38:73-89]. The plant roots stabilize the nest carton. AGs have been well-known in the neotropics for more than 100 years. In contrast, reports on similar associations in the paleotropics are scarce so far. After discovering a first common AG system on giant bamboo [Kaufmann et al. (2001) 48:125-133], we started a large-scale survey for AGs in Peninsular Malaysia, Borneo, Java, and southern Thailand. A great variety of AG systems (altogether including 18 ant species and 51 plant species) was discovered and is described in the present paper. The high number of species participating in AG associations was reflected by a great variability in the specific appearances of the nest gardens. Frequently, further groups of organisms (e.g., hemipteran trophobionts, fungi) were also involved. Preference patterns of particular ant and epiphyte species for each other and for particular phorophytes (carrier trees) were detected. We integrate domatia-producing, so-called ant-house epiphytes in our study and compare their phases of establishment, as well as other characteristics, to "classical" AGs, coming to the conclusion that they should be regarded only as a special type of AG epiphyte and not as a separate ecological category.