METHODS: Raw and cooked extracts of the giant freshwater prawn were prepared. The IgE reactivity pattern was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting technique with the sera of 20 skin prick test (SPT) positive patients. The major allergen identified was then characterized using the proteomics approach involving a combination of two-dimensional (2-DE) electrophoresis, mass spectrometry and bioinformatics tools.
RESULTS: SDS-PAGE of the raw extract showed 23 protein bands (15-250 kDa) but those ranging from 40 to 100 kDa were not found in the cooked extract. From immunoblotting experiments, raw and cooked extracts demonstrated 11 and 5 IgE-binding proteins, respectively, with a molecular mass ranging from 15 to 155 kDa. A heat-resistant 36 kDa protein was identified as the major allergen of both extracts. In addition, a 42 kDa heat-sensitive protein was shown to be a major allergen of the raw extract. The 2-DE gel fractionated the prawn proteins to more than 50 different protein spots. Of these, 10 spots showed specific IgE reactivity with patients' sera. Matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis led to identification of 2 important allergens, tropomyosin and arginine kinase.
CONCLUSIONS: It can be concluded that the availability of such allergens would help in component-based diagnosis and therapy of prawn allergies.
METHODS: The PCR technique was established based on published information and validated using blood sample of laboratory animals of which their whole gene sequences are available in GenBank. PCR was next performed to compile gene sequences of different species of wild rodents. The primers used were complementary to the conserved region of the cytb gene of vertebrate's mtDNA. A total of 100 blood samples, both from laboratory animals and wild rodents were collected and analyzed. The obtained unknown sequences were compared with those in the GenBank database using BLAST program to identify the vertebrate animal species.
RESULTS: Gene sequences of 11 species of wild animals caught in 9 localities of Peninsular Malaysia were compiled using the established PCR. The animals involved were Rattus (rattus) tanezumi, Rattus tiomanicus, Leopoldamys sabanus, Tupaia glis, Tupaia minor, Niviventor cremoriventor, Rhinosciurus laticaudatus, Callosciurus caniseps, Sundamys muelleri, Rattus rajah and Maxomys whiteheadi. The BLAST results confirmed the host with exact or nearly exact matches (>89% identity). Ten new gene sequences have been deposited in GenBank database since September 2010.
CONCLUSIONS: This study indicates that the PCR direct sequencing system using universal primer sets for vertebrate cytb gene is a promising technique for blood meal identification.
METHODS: Dietary intake of vitamins was assessed by 131 food frequency questionnaire items in both hypertensive participants and normotensive age-sex matched healthy controls. The associated changes in serum antioxidants and lipid peroxidation were also assessed along with lipid profile and anthropometric measurements in both groups of subjects under study.
RESULTS: Dietary vitamins intake was higher in hypertensive participants excepting for vitamin B2 and ascorbic acid compared to normotensive controls. Anthropometric variables in the hypertensive showed significant differences in weight, body mass index, waist circumference, hip circumference, waist-hip ratio and mid-arm circumference. The total cholesterol, low-density lipoprotein cholesterol, triglyceride were significantly higher (P<0.001) in hypertensive except high-density lipoprotein cholesterol which was significantly higher (P<0.001) in normotensive. The serum endogenous antioxidants and enzyme antioxidants were significantly decreased in hypertensive except serum albumin levels compared to normotensive along with concomitant increase in serum lipoprotein (a) malondialdehyde and conjugated diene levels.
CONCLUSIONS: Based on the observations, our study concludes that hypertension is caused due to interplay of several confounding factors namely anthropometry, lipid profile, depletion of endogenous antioxidants and rise in oxidative stress.