SLE is an autoimmune and polygenic disorder characterized by an accumulation and deposition of immune complexes. Several studies have indicated differential impact of FcgammaR polymorphism genotypes in different ethnic groups studied. The Fc receptor for IgG class IIA gene (FcgammaRIIA) occurs in two allelic forms. The allele FcgammaRIIA-H131 encodes a receptor with a histidine at the 131 amino acid position; the other allele FcgammaRIIA-R131 encodes an arginine. This polymorphism is believed to determine the affinity of the receptor for hIgG2 in immune complexes. FcgammaRIIA-H131 has a higher capacity for hIgG2 compared to FcgammaRIIA-R131 as measured by in vitro studies of insoluble immune complex clearance. We have investigated the polymorphism for FcgammaRIIA using a novel polymerase chain reaction-allele specific primer (PCR-ASP) method designed specifically to distinguish the two allelic forms. Our studies were based on 175 Chinese and 50 Malays SLE patients as well as 108 and 50 ethnically matched healthy controls for the respective groups. Analysis of the data (chi2 test with Yates correction factors and odds ratios) revealed that there were no significant differences between SLE patients and controls. We have not found evidence of a protective effect conferred by FcgammaRIIA-H131 in the ethnic groups studied.
This work represents the first application of the amplified fragment length polymorphism (AFLP) technique and the random amplified polymorphic DNA (RAPD) technique in the study of genetic variation within and among five geographical populations of M. nemurus. Four AFLP primer combinations and nine RAPD primers detected a total of 158 and 42 polymorphic markers, respectively. The results of AFLP and RAPD analysis provide similar conclusions as far as the population clustering analysis is concerned. The Sarawak population, which is located on Borneo Island, clustered by itself and was thus isolated from the rest of the populations located in Peninsular Malaysia. Both marker systems revealed high genetic variability within the Universiti Putra Malaysia (UPM) and Sarawak populations. Three subgroups each from the Kedah, Perak, and Sarawak populations were detected by AFLP but not by RAPD. Unique AFLP fingerprints were also observed in some unusual genotypes sampled in Sarawak. This indicates that AFLP may be a more efficient marker system than RAPD for identifying genotypes within populations.
Salbutamol ¿2-(tert-butylamino)-1-[4-hydroxy-3- (hydroxymethyl)phenyl]ethanol¿, also known as albuterol, is clinically the most widely used beta 2-adrenoceptor agonist in the treatment of bronchial asthma. During this study, we evaluated liquid-liquid extraction (LLE) and solid-phase extraction (SPE) in order to develop a reliable extraction method followed by analysis using liquid chromatography and gas chromatography. An assay is described which involves SPE as the clean-up method followed by gas chromatography-mass spectrometry to determine salbutamol levels in human serum after oral administration. The SPE method requires the use of a hyper-cross-linked styrene-divinylbenzene bonded phase (ENV+) without involving any sample pre-treatment to obtain 60-65% recoveries for salbutamol and terbutaline as the internal standard. Distilled water and 1% trifluoroacetic acid in methanol were found to be the most suitable washing solvent and eluting solvent, respectively. A detection limit of 2 ng mL-1 was achieved by derivatization with N-methyl-N-trimethylsilyltrifluoroacetamide to form trimethylsilyl (TMS)-salbutamol (m/z 369) and TMS-terbutaline (m/z 356). The relationship between the ratio of the peak area of salbutamol to that of the internal standard and concentration was linear for the range tested (2-200 ng mL-1) and the correlation of coefficient was 0.9999 with a y-intercept not significantly different from zero. The inter-day relative standard deviation (RSD) was < 10% for all three concentrations. The intra-day RSD was 14% for 2 ng mL-1. This assay was then successfully applied to human serum samples obtained from clinical trials after oral administration of salbutamol.
Isolation and polymerase chain reaction (PCR) were performed for detection of Mycoplasma pneumoniae from respiratory tract specimens obtained from 200 adult and 200 pediatric patients. M. pneumoniae was isolated from bronchoalveolar lavage fluid of 1(0.5%) adult patient and 4(2.0%) tracheal aspirates of pediatric patients. PCR was positive for only one (0.5%) broncoalveolar lavage fluid of an adult patient and fifteen (7.5%) tracheal aspirates of pediatric patients. This study suggested that M. pneumoniae was more frequently detected in pediatric patients and PCR appears to have advantages over isolation, in terms of rapidity and sensitivity.
Central nervous system manifestations are probably the most frequent extrapulmonary complications of infections due to Mycoplasma pneumoniae, occur mostly in children. In this study, we attempted to isolate M. pneumoniae and to detect the organism by polymerase chain reaction (PCR) from cerebrospinal fluid samples (CSF) of pediatric patients. Of the 244 CSF samples, no M. pneumoniae was isolated. Six (2.5%) of the CSF samples were positive by PCR amplification. More effort are necessary to isolate the organism from CSF samples in order to ascertain the role of M. pneumoniae in causing neurological complications.
FcgammaRIIs are the most widely distributed of the Fcgamma receptor family and play an important role in the clearance of immune complexes. Evidence that the FcgammaRIIa-R131 allotype is less able to process and clear immune complexes effectively suggests that this may be a disease susceptibility factor for systemic lupus erythematosus (SLE). Data from studies published thus far do not agree on the potential role of FcgammaRIIa polymorphism in the genetics of SLE. Most studies in fact show no evidence for any correlation between polymorphism of FcgammaRIIa and risk for SLE. However, it remains to be determined whether FcgammaRIIa polymorphism may play a critical role in certain groups of patients, especially in those of differing ethnic background. Polymorphism of FcgammaRIIa may also be important in determining disease phenotype, and identification of this influence may have important implications in patient care and in identifying patients for more aggressive therapy.
Physico-chemical and sensory characteristics of frankfurters prepared with three types of palm fats (PF60: 40, PF70: 30 and PF80: 20) and palm olein (POo) at 20 and 25% of fat levels were studied. Incorporation of different fats at 20 and 25% did not affect the cooking yields of the frankfurters. Frankfurters incorporated with 25% POo showed the highest value of water-holding capacity (WHC) among eight formulations. The frankfurters containing POo showed the least cooking loss compared to those with palm fats. The incorporation of different type and level of fats resulted in significant changes in the colour (lightness, redness, yellowness) of frankfurters. Texture profiles of both raw and cooked frankfurters were found to be altered by the blending of different type and level of fats. In raw frankfurters, hardness for frankfurters mixed with palm fats were significantly higher than the one with POo but greater values for cohesiveness was observed in raw frankfurters blended with POo. Lowest chewiness was demonstrated by frankfurters mixed with 20% POo. Grilling increased the hardness values of all frankfurters. Contrary to the raw counterparts, cooked frankfurter with POo was the hardest among all formulations. Cohesiveness and chewiness was also found to be significantly higher for cooked frankfurters mixed with POo. Raw frankfurters with fat content of 25% showed greater value in hardness than those of 20%. However, there were no significant differences (P > 0.05) observed for all the texture profile attributes in cooked frankfurters due to fat levels. In sensory evaluation, frankfurters prepared with POo were found to be most acceptable by consumer panels as they scored the highest for hardness rating, chicken flavour, oiliness and overall acceptance attributes.
This study was conducted to determine the factors involved in predicting the onset of paraplegia in tuberculosis of the spine. A cross-sectional, case control review of 85 cases of spinal tuberculosis was conducted at the National Tuberculosis Centre in Kuala Lumpur. Sixty-nine of these cases were normal neurologically, whilst 16 cases experienced neural deficit. The data was analysed using backward logistic regression and Fishers exact probability test. The factors studied included symptoms and signs of spinal tuberculosis, common investigations for tuberculosis, and physical factors of the disease. Only the erythrocyte sedimentation rate (ESR) showed a significant difference between the neural deficit and neurologically normal groups. This suggests that the ESR may be a factor in predicting evolution of paraplegia in spinal tuberculosis. In addition, it was noted that a low proportion of patients had positive sputum smear results and bacterial culture growth for mycobacterium tuberculosis suggesting these tests are of limited value for tuberculosis of the spine.
A new and simple HPLC method using fluorescence detection was developed to determine 9-methoxycanthin-6-one, an active compound of Eurycoma longifolia Jack in rat and human plasma. The method entailed direct injection of plasma sample after deproteinization using acetonitrile. The mobile phase comprised acetonitrile and distilled water (55 : 45, v/v). Analysis was run at a flow rate of 1.0 ml/min with the detector operating at an excitation wavelength of 371 nm and emission wavelength of 504 nm. The method was specific and sensitive with a detection limit of 0.6 ng/ml and a quantification limit of approximately 1.6 ng/ml. The method was applied in a pilot pharmacokinetic/bioavailability study of the compound in rats. Less than 1 % of the compound was found to be absorbed orally.