Accurate diagnosis of human filarial infections still remains a problem for clinicians and co-ordinators of filariasis control programs. Diagnosis of filariasis is based on parasitological, histopathological, clinical and immunological approaches. No significant advances have been made for the first three approaches although some refinements in their use and interpretation of results have occurred. For the immunological approach, intradermal tests and antibody detection assays using crude parasite extracts generally lack specificity and/or sensitivity to discriminate between past and present filarial infections in humans. Antigen detection assays would therefore provide a more accurate indication of active filarial infections. Several monoclonal antibodies to various stages of lymphatic filarial parasites have been developed and appear potentially useful for filarial antigen detection.
Introduction: Diabetes mellitus (DM) is one of the top diseases that lead public health concern in Malaysia. It was believed to rise in number up to 4.5 million on cases by year 2020 based on the current figure. Momordica charantia Linn (MC), a climber belonging to family Cucurbitaceae, is well known in treating diabetic-related conditions. In earlier studies related to the hypoglycemic properties of MC mainly utilized the crude extract, which contain a mixture of bioactives (charantins, insulin-like peptides and alkaloids). Till now, there is no conclusive result on the major bioactives that play role in the hypoglycemic effect of MC and research regarding the charantin purification was not well established. Hence, the objectives of this study were to purify the charantin from MC and to characterize the purified charantin before further subjected to in vivo hypoglycemic study. Methods: The crude was first extracted from MC using ethanol as solvent via Soxhlet extraction following by a series of purification steps via washing, centrifugation, and C-18 cartridges. Results: The HPLC analysis showed that the charantin of purified extract after passing out from the cartridge exuded at 12.50 min with a concentration of 500 ppm, which is relatively 20 times higher than the crude extract (25 ppm). The structural properties of purified charantin were studied using FTIR and it showed strong peaks of carboxylic acids (2884 nm), alcohols (1023 nm) and diethyl ether (1114 nm) as compared
with the standard. The compound was reconfirmed in LC-MS analysis. The result displayed mass spectrum in positive mode indicates the presence of similar compound in the purified extract and standard charantin, as presented by ion m/z = 300. Conclusion: The charantin was successfully purified from MC and can act as a potent plant-based hypoglycemic agent for diabetes.
The chemoprophylactic use of diethylcarbamazine citrate at total oral doses of 15--180 mg/kg body weight was tested against subperiodic Brugia malayi infection in the leaf monkey (Presbytis melalophos). A total dose of 45 mg/kg body weight given over 9 days killed all developing infective larvae. Similarly, a total dose of 35 mg/kg body weight given over 7 days killed all fourth stage larvae. The minimum effective dose that prevents infection would be 5 mg/kg body weight daily for 7 days every month.
Two monoclonal antibodies (MAbs), one produced against Plasmodium falciparum (PF-IG8) and the other against P. cynomolgi (PC-IE12) schizont antigens were used in a sandwich ELISA for the detection of circulating plasmodial antigens in sera of patients infected with either P. falciparum, P. vivax or P. malariae. The mean +/- SD optical density (OD) values for the normal control group using PF-108 and PC-1E12 were 0.351 +/- 0.036 and 0.205 +/- 0.044, respectively. Mean OD values for the three infected groups were found to be significantly higher than those of the normal control group for both MAbs. However, ELISA values for individual serum specimens did not correlate with the level of parasitemia in the infected blood. Using a cut-off point of mean OD +/- 3 SD of the normal control group as indicating a positive reading, the specificity of this assay with both MAbs was 100%. The sensitivity of the assay using PF-1G8 was 95% while that obtained with PC-1E12 was 98%.
The objective of this study was to evaluate the precursors of acrylamide formation in sweet potato (SP) (Ipomoea batatas L. Lam) chips and to determine the effect of different types of vegetable oils (VOs), that is, palm olein, coconut oil, canola oil, and soya bean oil, on acrylamide formation. The reducing sugars and amino acids in the SP slices were analyzed, and the acrylamide concentrations of SP chips were measured. SP chips that were fried in a lower degree of unsaturation oils contained a lower acrylamide concentration (1443 μg/kg), whereas those fried with higher degree of unsaturated oils contained a higher acrylamide concentration (2019 μg/kg). SP roots were found to contain acrylamide precursors, that is, 4.17 mg/g glucose and 5.05 mg/g fructose, and 1.63 mg/g free asparagine. The type of VO and condition used for frying, significantly influenced acrylamide formation. This study clearly indicates that the contribution of lipids in the formation of acrylamide should not be neglected.
3,4',5-Trihydroxy-3',7-dimethoxyflavanone was isolated from the ligroin extract of the leaves of Blumea balsamifera, while the acetone extract yielded 3',4',5-trihydroxy-7-methoxyflavanone and a new biflavonoid identifed as 3-O-7''-biluteolin (1). The isolation of 1 is significant since a biflavonoid with a C-O-C linkage of the type [I-3-O-II-7] has not been previously reported from a plant.
Transforming growth factor-beta (TGF-beta) has been shown to inhibit the growth of mammary epithelial cells and may play a protective role in mammary carcinogenesis. In contrast, oestrogens promote the development of breast cancer. Oestrone sulphate (E1S) is a huge reservoir of active oestrogens in the breast being converted to the weak oestrogen, oestrone (E1), by oestrone sulphatase. E1 is reversibly converted by oestradiol-17beta hydroxysteroid dehydrogenase to the potent oestrogen, oestradiol (E2). The aim of this study was to assess the effect of the TGF-beta1 isoform on growth and oestrogen metabolism in the hormone-dependent MCF-7 and hormone-independent MDA-MB-231 human breast cancer cell lines. The results showed that TGF-beta1 significantly inhibited cell growth and stimulated the conversion of E1S to E1 and E1 to E2 in the MCF-7 cell line. In the MDA-MB-231 cell line TGF-beta1 significantly stimulated cell growth and inhibited the interconversions between E1 and E2. In conclusion, the growth inhibitory effect of TGF-beta1 on the MCF-7 cell line would appear to confer a protective effect in breast cancer. However, its ability to increase the amount of E2 would increase the risk of breast cancer. Which of these effects predominates in vivo remains to be explored. The growth stimulatory effect of TGF-beta1 on the MDA-MB-231 cell line probably acts through a mechanism independent of the effect of TGF-beta1 on oestrogen concentrations since this cell line is hormone unresponsive.
Oestrogens play an important role in the development of breast cancer. A very important source of active oestrogens in the breast is oestrone sulphate which is converted to oestrone by oestrone sulphatase. The aim of this study was to assess the effects of IGF-I and IGF-II on oestrone sulphatase activity in, as well as cell growth of, MCF-7 and MDA-MB-231 human breast cancer cell lines. Cells were grown in supplemented DMEM and treated with varying concentrations of IGFs. At the end of the treatment period, intact cell monolayers were washed and assayed for oestrone sulphatase activity and the number of cell nuclei determined on a Coulter Counter. Oestrone sulphatase activity was significantly stimulated by IGF-I and II at concentrations of 100 ng/ml and 200 ng/ml in MCF-7 cells. IGF-I had no effect on oestrone sulphatase activity in MDA-MB-231 cells over the range of concentrations tested. Significant inhibition of oestrone sulphatase was observed in MDA-MB-231 cells at higher concentrations of IGF-II (50 ng/ml, 100 ng/ml and 200 ng/ml). Both IGF-I and IGF-II at higher concentrations (100 ng/ml and 200 ng/ml) significantly inhibited MCF-7 and stimulated MDA-MB-231 cell growth. Since IGF-I and II have effects on cell growth and oestrone sulphatase activity in breast cancer cell lines they may play a role in the development and progression of human breast cancer.
Edible bird nests (EBNs) are consumed worldwide for various health benefits. EBNs are nests built from the saliva of swiftlets of Aerodramus species. The global market for EBNs is on the rise, especially from Hong Kong and mainland China. In the past, EBNs were harvested mainly from natural caves; however in the recent years, there has been a rapid growth of swiftlet farming. Little is known about the actual composition of EBNs except for protein, carbohydrate, ash and lipid contents, amino acids, vitamins and macro/ micronutrients. Besides the biochemical components of EBNs, are there any other structures that are associated with EBNs? This paper reports on the structural analysis of raw unprocessed farm and processed commercial EBNs. The raw EBNs were purchased from swiftlet farms in five locations in Peninsula Malaysia: Kuala Sanglang (Perlis; 6° 16' 0"N, 100° 12' 0"E), Pantai Remis (Perak; 4º 27' 0" N, 100º 38' 0" E), Kluang (Johor; 02º 012 303N 103º 192 583E), Kajang (Selangor; 2º 59' 0"N, 101º 47' 0"E) and Kota Bharu (Kelantan; 6º 8' 0"N, 102º 15' 0"E). The commercial nests were purchased from five different Chinese traditional medicinal shops (Companies A-E). A portion of each EBN was randomly broken into small fragments, attached to carbon tape and coated with gold and palladium particles for examination and photography under a scanning electron microscope. Structural analysis revealed the presence of mites, fungi, bacteria and feather strands on both the raw and commercial nests. Mite eggshells and faecal pellets, and body parts of other arthropods were seen only in the raw nests. The commercial nests had a variety of unidentified structures and substances coated on the nests' surfaces that were not found on the raw nests. The presence of these contaminants may jeopardise the quality of EBNs and pose health risks to consumers. Further identification of the mites and their allergens, fungi and bacteria are on-going and will be reported separately.
Biotechnological tools are being used in malaria, filariasis and dengue research. The main emphasis has been on the production of reagents for immunodiagnosis and research. In this respect monoclonal antibodies (McAbs) against various species and stages of the above pathogens have been produced. It is hoped that these McAbs will be useful not only in immunodiagnosis but also for seroepidemiological applications. A DNA probe against Brugia malayi has been tested in Malaysia and was found to be sensitive and specific.
Comparative studies of vector efficiency were done with the Liverpool and Malaysian strains of Aedes (Finlaya) togoi for subperiodic Brugia malayi and Brugia pahangi. The Malaysian strain of A. togoi was found to take in fewer microfilariae under the same experimental conditions than the Liverpool strain. Also, for various microfilarial densities in the host's peripheral blood, the Malaysian strain had less mean infective larvae per fed mosquito than the Liverpool strain. The microfilarial intake of A. togoi was not affected by the site of feeding on the host affected by the site of feeding on the host. Most of the mosquitoes took in fewer microfilariae than expected. It is concluded from these studies that the Malaysian strain of A. togoi is a susceptible and reasonably good vector for subperiodic B. malayi and B. pahangi. Further field studies should be carried out to determine its importance as a natural vector of Brugian filariasis.
Human toxocariasis which is caused mainly by the larvae of Toxocara canis and Toxocara cati, is a worldwide zoonotic disease that can be a potentially serious human infection. The enzyme-linked immunosorbent assay (ELISA) using T. canis excretory-secretory (TES) antigens harvested from T. canis larvae is currently the serological test for confirming toxocariasis. An alternative to producing large amounts of Toxocara TES and improved diagnosis for toxocariasis is through the development of highly specific recombinant antigens such as the T. canis second stage larva excretory-secretory 30 kDa protein (recTES-30). The aim of this study was to evaluate the sensitivity and specificity of a rapid diagnostic kit (RDT, named as iToxocara kit) in comparison to recTES-30 ELISA in Serendah Orang Asli village in Selangor, Malaysia. A total of 133 subjects were included in the study. The overall prevalence rates by ELISA and RDT were 29.3% and 33.1%, respectively, with more positive cases detected in males than females. However, no association was found between toxocariasis and gender or age. The percentage sensitivity, specificity, positive predictive value and negative predictive value of RDT were 85.7%, 90.1%, 80% and 93.2%, respectively. The prevalence for toxocariasis in this population using both ELISA and RDT was 27.1% (36/133) and the K-concordance test suggested good agreement of the two tests with a Cohen's kappa of 0.722, P<0.01. In addition, the followed-up Spearman rank correlation showed a moderately high correlation at R=0.704 and P<0.01. In conclusion, the RDT kit was faster and easier to use than an ELISA and is useful for the laboratory diagnosis of hospitalized cases of toxocariasis.
The dynamics of the transmission of subperiodic Brugia malayi in a typical endemic area in Malaysia was studied over a period of 4 years. Mass chemotherapeutic control with diethylcarbamazine citrate was found to be inefficient, new cases being detected even after the fifth treatment cycle of 6 mg/kg X 6 days per cycle. This is in marked contrast to the situation in periodic b. malayi areas where mass treatment efficiently controlled the infection. The disparity in results in these two areas is attributed to zoonotic transmission of subperiodic B. malayi from non-human primates where a mean infection rate of 76.3% was found.
Malarial antibodies in 80 patients were measured using the diffusion-in-gel enzyme linked immunosorbent assay (DIG-ELISA), enzyme-linked immunosorbent assay (ELISA) and the indirect fluorescent antibody (IFA) test. Good correlations were obtained between all three tests in terms of sensitivity and reliability. DIG-ELISA has the advantage of being a rapid diagnostic tool for the detection of malarial antibodies.
In spite of more than 30 years of control activities, malaria continues to be the most important parasitic infection in Malaysia, accounting for 39,189 confirmed cases in 1991, giving an annual parasite incidence rate of 2.2 per 1,000 population. Some factors contributing to the continued transmission of malaria are the development of drug resistant Plasmodium falciparum, changes in vector behavior, and ecological changes due to socio-economic reasons. Malaria parasite rates are higher among the Aborigines, land scheme settlers and those in intimate contact with the jungle, like loggers. There has been no substantial change in the proportion of the three common malaria species responsible for infections, P. falciparum, P. vivax, P. malariae and mixed infections accounting for about 70%, 28%, 1% and 1%, respectively of all infections. Drug resistant P. falciparum is unevenly distributed in Malaysia, but based on clinical experience and in vitro drug sensitivity studies, chloroquine resistance is frequently encountered. There has been clinical and laboratory evidence of resistance to sulfadoxine/pyrimethamine combination as well as quinine, but all these have so far been successfully treated with a combination of quinine and tetracycline. The eradication of the disease is impossible in the near future but there is confidence that with better surveillance techniques and the use of alternative control measures like permethrin impregnated bed-nets to complement existing ones, the target of bringing down the annual parasite incidence to 2 per 1,000 population during the Sixth Malaysian Plan period (1991-1995) can be achieved.
Mass drug administration via 3 modes of delivery reduced the incidence and prevalence rates and intensity of Brugia malayi infection in 3 rural villages in the Bengkoka Peninsula, Sabah, in 1982-1983. A dosage of 6 mg diethylcarbamazine citrate (DEC-C)/kg body weight was administered either daily or weekly (total of 6 doses, 36 mg/kg body weight), and impact on B. malayi cases were comparable in the 3 villages. A total of 384 people participated in the DEC-C regimens, and all pregnant women and children under 2 years were excluded from the study. Bekessy's method of estimation of incidence and recovery rates was applied to data on B. malayi microfilaremia before drug administration. Treatment with DEC-C by any of the 3 modes of delivery drastically reduced the number of episodes of patent microfilaremia, incidence and prevalence, and median microfilarial density. Reduction was sustained for at least 18 to 24 months after treatment.
Studies on parasite populations in Antarctic soils are scarce and thus little is known about the threat of these parasites towards either the natural fauna or human visitors. However, human presence in Antarctica, mainly through research and tourism, keeps increasing over time, potentially exposing visitors to zoonotic infections from Antarctic wildlife and environment. Most available literature to date has focused on faecal samples from Antarctic vertebrates. Therefore, this study addressed the possible presence of parasites in Antarctic soil that may be infectious to humans. Soil samples were obtained from five locations on Signy Island (South Orkney Islands, maritime Antarctic), namely North Point and Gourlay Peninsula (penguin rookeries), Pumphouse (relic coal-powered pump house), Jane Col (barren high altitude fellfield) and Berntsen Point (low altitude vegetated fellfield close to current research station). Approximately 10% of the soil samples (14/135) from 3 out of the 5 study sites had parasites which included Diphyllobotridae spp. eggs, Cryptosporidium sp., an apicomplexan protozoa (gregarine), Toxoplasma gondii, helminths (a cestode, Tetrabothrius sp., and a nematode larva) and mites. The presence of parasites in the 3 sites are most likely due to the presence of animal and human activities as two of these sites are penguin rookeries (North Point and Gourlay Peninsula) while the third site (Pumphouse Lake) has human activity. While some of the parasite species found in the soil samples appear to be distinctive, there were also parasites such as Cryptosporidium and Toxoplasma gondii that have a global distribution and are potentially pathogenic.