Over the past year, many reports have been published on a variety of clinical manifestations related to antiphospholipid antibodies. The low prevalence of anticardiolipin antibodies with the rare occurrence of thrombosis and a low rate of fetal loss in studies in Malaysia and China showed a potential role for local factors. A study of cross-reactive idiotype of the anticardiolipin antibody suggested that anticardiolipin antibodies are derived from a set of natural autoreactive clones. Regarding the pathogenic role of the antiphospholipid antibody, evidence has been presented that the epitopes formed between cardiolipin and beta 2 glycoprotein I are the targets of the antiphospholipid antibody. Complement activation, abnormalities of natural anticoagulants such as protein S deficiency, and genetic association with DR4, DR7, and DRw53 have also been studied.
Eighteen infants clinically suspected to be intolerant of cow's milk were placed on a milk-free formula and six to eight weeks later were orally challenged with cow's milk. Following challenge three groups were recognised. Group A: Four infants tolerated oral feeds ofcow's milk and lacked mucosal abnormality or clinical symptoms. Group B: Seven infants had mucosal deterioration but lacked clinical symptoms and tolerated cow's milk. Group C: Seven infants had mucosal abnormality, developed clinical symptoms and were intolerant of cow's milk. The intestinal transudation of IgA was increased in Group A and unchanged in Group Band C : the IgM levels in the duodenal juice was increased in Group A and B but unchanged in Group C : the IgG levels in the juice were increased in all Groups following challenge. It appears that increased transmission of IgA and IgM or IgM alone in the duodenal juice is associated with lack of development of clinical symptoms. Symptoms are present in infants in whom the IgA and IgM levels in duodenal juice remained unchanged after challenge. It is suggested that patients responding to cow's millt challenge with intestinal production of IgA and IgM (or IgM alone) are able to counter balance the deleterious mechanisms leading to clinical cow's milk intolerance whereas those who, for some unknown reason, do not mount a secretory immune response become ill.
Fifteen independent E. coli strains of avian, bovine and porcine origin in Peninsular Malaysia were tested for antibiotic resistance and conjugative R plasmids. Eight (53%) isolates were found to be antibiotic resistant. Among them, 37.5% were mono-resistant and 62.5% were resistant to three or more antibiotics, i.e., multi-resistant. All of them were resistant to Tc and sensitive to Gm and Nx. Three of the eight antibiotic resistant strains were able to transfer all or part of their resistance to an E. coli K12 recipient by conjugation. The transfer frequencies of Km, Sm and Tc resistance of the three donors varied between 4.5 X 10(-8) to 6.8 X 10(-7). Analysis of the plasmid profiles of all the three donors and their respective transconjugants after agarose gel electrophoresis provided conclusive evidence that the transferable resistance traits were plasmid-mediated.
The effects of cell density and growth in 10% foetal bovine serum and 10% newborn calf serum on the activity of the enzyme (Na+ + K+)-ATPase were studied in 3T3 and SV3T3 cells. The enzyme activity decreases in 3T3 cells grown in foetal bovine serum as the cells approach confluency while in those grown in newborn calf serum the enzyme activity increases. The (Na+ + K+)-ATPase activity does not change with increase in cell density in SV3T3 cells grown in foetal bovine serum while the enzyme activity in those grown in newborn calf serum increases with increase in cells density up to about 1.35 x 10(5) cells/sq. cm. and then decreases with further increase in cell number. At confluency it was found that the enzyme activity is higher in the SV3T3 as compared to the 3T3 cells when the cells were grown in 10% foetal bovine serum, whereas in those grown in 10% newborn calf serum the enzyme activity is higher in the 3T3 as compared to the SV3T3 cells.
Current research is based on the synthesis of novel (E)-4-aryl-2-(2-(pyren-1-ylmethylene)hydrazinyl)thiazole derivatives (3-15) by adopting two steps route. First step was the condensation between the pyrene-1-carbaldehyde (1) with the thiosemicarbazide to afford pyrene-1-thiosemicarbazone intermediate (2). While in second step, cyclization between the intermediate (2) and phenacyl bromide derivatives or 2-bromo ethyl acetate was carried out. Synthetic derivatives were structurally characterized by spectroscopic techniques such as EI-MS, 1H NMR and 13C NMR. Stereochemistry of the iminic double bond was confirmed by NOESY analysis. All pure compounds 2-15 were subjected for in vitro β-glucuronidase inhibitory activity. All molecules were exhibited excellent inhibition in the range of IC50=3.10±0.10-40.10±0.90μM and found to be even more potent than the standard d-saccharic acid 1,4-lactone (IC50=48.38±1.05μM). Molecular docking studies were carried out to verify the structure-activity relationship. A good correlation was perceived between the docking study and biological evaluation of active compounds.
Stem cells from human extracted deciduous teeth (SHED) have the ability to multiply much faster and double their population in culture at a greater rate, indicating that it may be in a more immature state than other type of adult stem cells. Mesenchymal stem cells (MSC) from human primary molars were isolated and cultured in media supplemented with 20% fetal bovine serum. The MSCs were confirmed using CD 105 and CD 166 and the identification of the osteoblast cells were done using reverse transcriptase polymerase chain reaction (RT-PCR) analysis. Differentiated osteoblast cells (DOC) were characterized by alkaline phosphotase and von Kossa staining followed by immunocytochemistry staining using osteocalcin and osteonectin antibodies. Further validation of SHED was done by RT-PCR to detect the presence of insulin-like growth factor 2 (IGF-2) and discoidin domain tyrosine kinase-2 (DDTK-2) transcripts, while the presence of Runx-2 mRNA was used to characterize DOC. The results showed that SHED was found positive for CD 105 and CD 166 and could differentiate into osteoblast, bone forming cells. The findings revealed the presence of distinct MSC population which had the capability to generate living human cells that could be a possible source for tissue engineering in the future.
To gain insight into the microbiological safety of food products routinely traded across international borders in Southeast Asian countries, beef imported from Malaysia to southern Thailand was examined for contamination with Escherichia coli O157 and its subsequent spread into the imported areas. We screened 31 samples exported from Malaysia and 36 domestic Thai samples. Isolation methods including an O157 antigen-targeted immunomagnetic separation technique, screening on CHROMagar O157 medium, and serotype confirmation of E. coli isolates by specific agglutination tests were employed. Fourteen strains of E. coli O157:H7 were isolated from eight Malaysian samples (25.8%) and six strains from four Thai samples (11.1%). These strains were of the stx1-stx2+eae+ genotype except one Malaysian strain which was of the stx1- stx2- eae+ genotype. All 19 O157:H7 strains possessing the stx2 gene produced little or no Stx2 (reversed passive latex agglutination titer ≤ 4). Of the 19 strains, five Malaysian (38.5%) and two Thai (33.3%) strains exhibited resistance to a set of antibiotics. Finally, the results of two DNA fingerprinting
analyses (O157 IS-printing targeted to IS629 and pulsed-field gel electrophoresis, PFGE) of the O157:H7 strains possessing the stx2 gene, indicated that the Malaysian and Thai strains are closely related. Therefore, E. coli O157:H7 might be transferred from Malaysia to southern Thailand through beef trade.
Three restriction enzymes were used in Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) using the mitochondrial cytochrome b region to establish a differential diagnosis which detect and discriminate between three meat species: pork, cow and chicken. DNA was extracted from samples containing meat of a single animal such as raw pork (Sus scrofa domesticus), chicken (Gallus gallus) and cow (Bos taurus) as well as mixed samples of two species of animals in different ratios. The amplified 359 base pairs (bp) portion of the mitochondrial cyt b gene from pure or mixed samples in different ratios was cut using three different restriction enzymes resulting in species specific restriction fragment length polymorphism (RFLP). This technique proved to be extremely reliable in detecting the presence of low levels of target DNA obtained from a 0.25 mg component in a particular mixed meat sample. This revealed the cyt b region as highly conserved and consequently a good molecular marker for diagnostic studies. Thus, this technique can be applied to food authentication for the identification of different species of animals in food products.
A study was carried out to distinguish mono- (MAG) and di-acylglycerol (DAG) from plant lipids such as sunflower, rapeseed and soybean oil, from those derived from animal fats such as lard, goat fat and beef fat using fatty acid and thermal profile data. MAG and DAG of both plant and animal lipids were synthesized according to a chemical glycerolysis method catalyzed by sodium hydroxide. MAG and DAG of individual lipid were isolated and purified using the standard column chromatography method and subjected to fatty acid analysis by gas chromatography (GC) and thermal analysis by differential scanning calorimetry (DSC). The application of principal component analysis (PCA) to the data collected from the individual instrumental technique showed that it was possible to distinctly classify MAG and DAG of plant lipids from those derived from animal fats.
Forty three (n=43) genomic DNA of Escherichia coli (11 isolates from eggs and 32 isolates from imported beef meats) were characterized by shiga toxin 1 (stx1), enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) and random amplified polymorphic DNA-PCR (RAPD-PCR) analyses. In the shiga toxin 1 (stx1) gene detection with primer stx 1F (5’-TTCTTCGGTATCCTATTCCC-3’) and stx 1R (5’- CTGTCACAGTAACAACCGT-3’), 9 E. coli of beef meats isolates were positive toward sxt1 gene. The results of the ERIC-PCR and RAPD-PCR were analyzed using GelCompar II software. ERIC-PCR with primer ERIC1 (5’-CACTTAGGGGTCCTCGAATGTA -3’) and ERIC2 (5’-AAGTAAGTGACTGGGGTGAGCG-3’) discriminated the E. coli into 6 clusters and 10 single isolates at 80% similarity. RAPD-PCR with primer Gen8 and Gen9, produced 10 clusters and 15 single isolates and 12 clusters and 14 single isolates of 80%, respectively. These results demonstrated that both ERIC-PCR and RAPD-PCR are useful and suitable tools for molecular typing of those isolates examined.
Legislation concerning the safety assessment and labelling of foodstuffs has been implemented in many countries. Consequential to a number of cases of food adulteration reported globally, a fast and reliable detection method for the food traceability is required in ensuring effective implementation of food legislation in a country. In this study, PCR-RFLP technique based on cyt b gene has been tested for its suitability for these purposes. This method combines the use of a pair of universal primer that amplifies a 359 bp fragment on the cyt b gene from meat muscle DNA and restriction enzyme analysis. Analysis of experimental beef frankfurter, minced beef, pork frankfurter and pork cocktail samples demonstrated the suitability of the assay for the detection of the beef (Bos taurus) and pork (Sus scrofa), but not applicable for some processed food, particularly detection of mackerel (Rasterelliger brachysoma), sardine (Saedinella Fimbriata) and tuna (Thunnus tonggol) origin in canned food. Commercial frauds through species mislabelling or misdescribed were not detected. The assay is demonstrated applicable for routine analysis of meat traceability of foodstuffs and legislation purposes, if sufficient availability of detectable mtDNA in the foodstuffs is ensured.
There are disturbing trends of emerging and re-emerging infectious diseases, globally and locally thus giving it a real cause of concern. These include diseases by agents hitherto unknown in human such as Severe Acute Respiratory Syndrome (SARS) and Ebola; those that were purely zoonoses but had now affected man such as bovine spongioform encephalitis (BSE) in United Kingdom and avian influenza in Vietnam and Thailand; those that were thought to be eliminated but reappeared such as plague in India and those that begun to show reverse trends such as tuberculosis (TB) and malaria. Malaysia is no exception. Viral fever EV71, Chinkugunya, Conjunctivitis C24 variant or Nipah encephalitis were local examples of unknown or exotic infectious diseases occurring in recent years. In this age of globalization with expanding air travel and industrial trade, Malaysia is vulnerable to a wide array of new and resurgent infectious diseases. Apart from the direct health consequences on morbidity, mortality and its staggering cost, these infections also have far reaching implications upon sustainable development, psychosocial, economic, political and global security.
Protein Efficiency Ratio (PER) is the most widely used method for determining protein quality. The studies involved a few category of products as raw materials namely poultry products, beef burger products, fish and fish products, soy products and palm kernel cake in animal diet preparation were compiled to compare the data. Data from the proximate analysis showed that protein content in soy protein isolate (SPI) was the highest (95.00%) followed by meat such as mackerel fish (89.09%) and beef (88.60%). Results from feed consumption and total protein consumed showed that the rats fed with mechanically deboned poultry meat (MDPM) products (excluding broiler back) consumed more feed, ranging from 469.2g to 422.3g during the study while the lowest total feed consumed (157.7g) was recorded in the rat fed a diet of fermented palm kernel cake (fPKC). The total protein consumed by rat for diets of fish and fish products such as canned sardine was 62.36g, mackerel 61.76g and anchovy at 58.91g, followed by MDPM products. Tempeh (14.72g) and fPKC diet (16.3g) were among the lowest total protein consumed by the rats. Growth and PER data for rats fed a diet of canned sardine, anchovy and mackerel, as well as mechanically deboned turkey meat (MDTM) and mechanically deboned chicken meat (MDCM) had higher mean body weight (154.80g, 145.20g, 144.81g, 148.7g and 142.5g respectively) compared to rats fed with plant protein diet such as SPI, tempeh and PKC (34.79g, 16.34g and 16.60g respectively) whereas rats fed diets containing fPKC had a mean body weight loss of 24.4g. MDPM showed higher PER value (ranging from 3.01 to 3.34) compared to hamburger group, pure beef and fish group. Tempeh and SPI had lower PER of 1.02 and 1.52 respectively while the lowest PER of 0.50 and -1.50 were shown in PKC and fPKC. The highest digestibility was shown in mackerel (96.99%), followed by canned sardine (96.88%), tempeh (91.41%), meat (90.79%) and pure beef burger (90.04%) while digestibility of PKC and fPKC were much lower (45.70% and 22.60%). Lipid profile of rats fed with palm based fat beef burger showed that palm fat(PF) and red PF did not affect the total cholestrol concentration but resulted in higher high density lipoprotein (HDL)- cholesterol concentration in their blood serum. In summary, the utilization of PF and red PF in beef burger increased the HDL-cholesterol and has no effect on the concentration of total cholesterol in rat blood serum.
The objective of this study was to examine the effect of surfactant replacement therapy on hospital resource uti-lization in a well defined cohort of preterm infants with respiratory distress syndrome (RDS). Thirty preterm infants 800g with RDS requiring mechanical ventila-tion with Fi02 of 0.4 given modified bovine surfactant (Survanta) were compared with an equal number of his-torical controls. The total cost of neonatal care was cal-culated in a detailed survey covering all aspects of resource use. Surfactant-treated infants had an improved survival rate (90.0% vs 66.7%, p=0.03) and a trend towards shorter ventilator days (11.8 vs 19.0 days, p=0.17). There were no significant differences in the number of laboratory and radiological investiga-tions, use of disposable items, equipment, medications and other therapies. The total hospital cost per livebirth for surfactant-treated and control infants were R/V120,281 and R1V121,785 respectively. Personnel salaries represented the largest sector of resource uti-lization. When analysed by birthweight categories, the cost per livebirth for surfactant-treated and control infants in the 800-999g category were RM37,315 and RM14,760 respectively. As for the surfactant-treated and control infants in the 1000-1499g category, the cost per livebirth were RM21,426 and RM32,327 respectively. We concluded that surfactant replacement therapy did not increase overall hospital resource uti-lization and may decrease the cost for infants weighing 1000g.
Natural preservatives having the great antioxidant and antimicrobial activity have been utilized in the food industry for many years. In the present study, the effect of of two brands of commercial Assam green tea infusion (represented by A and B) and 0.02% BHA/BHT on microbial growth, anti-lipid oxidation and color change were investigated in cooked beef. The green tea concentration has influenced to the results. It was found that A and B at the concentration of 250 mg/mL significantly reduced the population of Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium and E. coli in the cooked beef to an undetectable level within 2 days of storage at 4oC. A and B also exhibited higher anti-lipid oxidation activity compared to 0.02% BHA/BHT, and control. Assam green tea infusions in cooked beef significantly increased ∆ L*
value and decreased ∆ a* and ∆ b* value (p ≤ 0.05). These indicate that Assam green tea infusion might be a potential candidate as a natural preservative for beef and other types of food.
Escherichia coli and Escherichia coli O157 were identified from “selom” (Oenanthe stolonifera), “pegaga” (Centella asiatica), beef, chicken, lamb, buffalo, “ulam Raja” (Cosmos caudatus) and “tenggek burung” (Euodia redlevi). The bacteria were recovered using chromagenic agar. Isolated Escherichia coli and Escherichia coli 0157 were further characterized by plasmid profiling and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). The virulence genes of the isolates (VT1, VT2, LT, ST, eaeA, inV) that produces pathogenic Escherichia coli and 16S rRNA gene were screened by a multiplex PCR assay. The plasmid profiling analysis showed that out of 176 isolates, only 103 isolates contained plasmids. ERIC-PCR analysis generated amplified products in the range of ~150 bp to > 1000 bp categorizing isolates into a total of 52 different profiles. Multiplex PCR showed that 20 (32.3%) of the isolates carried eaeA gene, 6 (9.7%) isolates possessed inV genes, only 1 (1.6%) have VT2 genes and 1 (1.6%) as well carried VT1 genes, 2 (3.2%) of the isolates harboured LT genes, and only 1 (1.6%) isolate possessed ST genes. There were no correlation between plasmid, ERIC-PCR and virulence genes profiles.