High performance liquid chromatography (HPLC) with phenylisothiocyanate (PITC) is recently used for confirming the diagnosis of inborn errors of metabolism (IEM) especially amino acid disorders in Malaysian children. The method of HPLC used is a precolumn derivatization of amino acids with phenylisothiocyanate and is separated by reversed phase chromatography using 3.9 x 300 mm free amino acid columns and is detected by a UV/Vis detector. The samples are obtained from cases suspected of inborn errors of metabolism, especially of amino acid disorders, which are detected clinically by pediatricians. Initially, samples from patients suspected of inborn errors of metabolism, either urine or serum, are run on one-dimensional thin layer chromatography and supplementary chemical tests to detect the abnormal bands and associated abnormalities respectively. Positive samples are further run on HPLC to determine the specific amino acids abnormality. An examples of a case of maple syrup urine disease is discussed, based on the thin layer chromatography findings and HPLC findings.
Dentists can offer their patients who smoke tobacco assistance with smoking cessation. We conducted this study to assess dental patient knowledge about the effects of smoking and perceptions and attitudes regarding the role of dentists in smoking cessation counselling. We conducted this study to inform tobacco cessation programs that could potentially include dentists. We conducted a cross-sectional study using a self-administered questionnaire among 375 patients. The mean age of subjects was 33.4 years; females comprised 51.5%. Participants were divided into 3 groups: those who never smoked (n = 263, 70.7%), smokers (n = 92, 24.7%), and ex-smokers (n = 17, 4.5%). Significantly more participants (p = 0.046) who never smoked (92.9%) knew smoking can cause bad breath than smokers(86.9%). Significantly more participants (p = 0.002) who never smoked (74.8%) knew smoking can cause periodontal disease than smokers (57.6%). Significantly
more participants (p < 0.001) who never smoked (84.5%) knew smoking can cause
oral cancer than smokers (66.7%). Significantly more participants (p < 0.001) who never smoked (86.7%) knew smoking can cause lung cancer than smokers (69.7%). Significantly more participants who never smoked (85.5%) felt dentists should be interested in the smoking status of their patients (p = 0.004) than smokers (72.6%). Significantly more participants (p = 0.08) who never smoked (69.6%) stated dentists should give smoking cessation advice than smokers/ex-smokers (59.0%). Smoker/ex-smokers had less knowledge about the effects of smoking on oral and general health than non-smokers. Both smokers/ex-smokers and non-smokers felt dentists should provide smoking cessation advice.
Various studies on toxoplasmosis in Malaysia have shown that specific antibodies to Toxoplasma gondii are common among Malaysians. Among the ethnic groups, the Malays have the highest prevalence rate followed by Indians, Orang Aslis (aborigines) and Chinese. Antibody is acquired early in life and increases with age. There is no significant difference in the prevalence rate between males and females. The disease is apparently more prevalent among rural dwellers and those in the lower socioeconomic group. It appears that the prevalence rate is also influenced by environmental conditions, occupation, diet and cultural habits. Studies with animals have shown the presence of antibody to T. gondii, but this does not seem to be the source of infection since Malaysians normally cook their meat well.
IgE-mediated allergic reaction to squid is one of the most frequent molluscan shellfish allergies. Previously, we have detected a 36 kDa protein as the major allergen of Loligo edulis (white squid) by immunoblotting using sera from patients with squid allergy. The aim of this present study was to further identify this major allergen using a proteomics approach. The major allergen was identified by a combination of two-dimensional electrophoresis (2-DE), immunoblotting, mass spectrometry and bioinformatics tools. The 2-DE gel fractionated the cooked white squid proteins to more than 50 different protein spots between 10 to 38 kDa and isoelectric point (pI) from 3.0 to 10.0. A highly reactive protein spot of a molecular mass of 36 kDa and pI of 4.55 was observed in all of the patients' serum samples tested. Mass spectrometry analysis led to identification of this allergen as tropomyosin. This finding can contribute to advancement in component-based diagnosis, management of squid allergic patients, to the development of immunotherapy and to the standardization of allergenic test products as tools in molecular allergology.
The aim of this study was to identify the major allergens of wildflower honey in local patients with atopic disease. SDS-PAGE revealed ten protein bands of 25 to 110 kDa, with a heavy cluster in region of 40-75 kDa. Immunoblotting demonstrated seven IgE-binding bands of 39 to 110 kDa. The 60 kDa protein had the highest frequency of IgE-binding (100%) followed by 54 kDa protein (95%), thus identified as the major allergens of wildflowerhoney. Our findings indicate that the allergen extract used for diagnosis of honey allergy contains both the 54 kDa and 60 kDa proteins.
Allergy to different classes of mollusks, including squid, which are members of the class Cephalopods has been reported. Tropomyosin, a major muscle protein, is the only well-recognized allergen in squid. The aim of this study was to characterize IgE-binding proteins of local Loligo edulis (white squid) consumed in Malaysia. Protein profiles and IgE-binding proteins were detected by sodium dodecyl sulfate-polyacrylamide gel-electrophoresis (SDS-PAGE) and immunoblotting using sera from 23 patients with positive skin prick test to raw squid extract. SDS-PAGE of the raw extract exhibited 21 protein bands (10-170 kDa) but those ranging from 19 to 29 kDa and 41 to 94 kDa were not found in the cooked extract. Immunoblotting of raw extract demonstrated 16 IgE-binding bands, ranging from 13 to 170 kDa. A heat-resistant 36 kDa protein, corresponding to squid tropomyosin, was identified as the major allergen of both extracts. In addition, a 50 kDa heat-sensitive protein was shown to be a major allergen of the raw extract. Our findings indicate that the allergen extract used for diagnosis of squid allergy should contain both the 36 kDa and 50 kDa proteins.
Serum IgG levels and complement C3 levels were assayed on Day 0, 1, 3-4, 7 and 56-70 post-treatment with diethylcarbamizine citrate (DEC) in a series to 26 patients with Brugia malayi infection and 6 volunteers without infection. On treatment, the microfilariae were cleared from the blood within 24 hours. The eosinophils decreased dramatically on Day 1 post-treatment but increased rapidly by Day 4 to 7 and then dropped to normal levels in 45 days. The serum IgG mean levels decreased briefly following treatment with DEC but then returned to original levels. However, the complement C3 levels gradually increased over the 2 months period of study reaching statistical significance levels (p less than 0.01) in patients with initial high blood microfilariae. The observation suggests that Brugia malayi infection probably induces a high rate of synthesis of complement C3 and this process continued in the post-treatment phase. Since, DEC treatment did not cause a decrease in complement C3 with the elimination of blood microfilariae, it would appear that the complement C3 is consumed following antibody attachment to the microfilariae as they enter the blood circulation.
Serum tumor necrosis factor alpha (TNF) concentration was assayed in 105 patients with nasopharyngeal carcinoma using a sensitive ELISA technique with detection level of 10 pg/ml. The TNF levels were detectable in 45 of 63 (71.4%) patients newly diagnosed for the malignancy and 29 of 42 (69%) patients in remission following treatment with radiotherapy. In 25 normal controls the TNF were less than 10 pg/ml. While TNF may be present in the majority of the patients with the malignant disease, the TNF concentration appeared to have no clinical significance in diagnosis or prognosis of the patients.
The antibody titres to P. falciparum and Epstein-Barr Virus-associated antigens were assayed in 22 patients with NPC and 43 controls. All, but one patient had antimalarial titres; 14 had titres greater than 80 and 4 patients greater than 640. Compared to controls the mean anti-malarial titre for most age groups were higher in the patients. Those patients with high anti-malarial titres also had high IgA anti-VCA titre, an antibody which has been demonstrated to be diagnostic for NPC. The peak anti-VCA (IgG) and anti-EA (IgG) antibody titres were associated with anti-falciparum titres of 320-640 and 80-160, respectively. The results are discussed in relation to the possible association between malarial infection and etiology of NPC.
The prevalence of antibody to human herpesvirus type 6 (HHV-6) and Epstein-Barr virus (EBV) viral capsid antigens (VCA) were analysed in sera from Kadazans of Sabah, North Borneo. At a serum dilution of 10, about 34% were positive for HHV-6 antibody but in contrast all 95 individuals studied were positive for EBV VCA antibody. The study shows that HHV-6 and EBV infection occur independently. The low frequency of seropositive individuals in this community suggests that other than socioeconomic factors are responsible for the spread of the virus.
Serum immunoglobulin G, A, M, D and E levels were determined in the forest-dwelling Orang Asli of age group 8 to 64 years. The levels are higher than observed for urban Malaysians and comparable to levels reported for populations residing in the tropics. There was no significant difference in serum levels of all the immunoglobulins studied in both sexes. The elevated serum immunoglobulins levels are discussed in terms of the nature of the immune defence developed in the Orang Asli to contend with the many parasites prevalent in their environment.
A questionnaire survey to study the smoking habits and attitudes toward smoking among secondary school teachers in Kelantan, Malaysia was conducted between July and September 1992. Questionnaires were sent to 5,112 teachers through their respective headmasters. Sixty-three percent (3,208 teachers; 61% males, 39% females) responded satisfactorily. Overall, 625 teachers (20%) were current smokers, 141 (4%) were occasional smokers, 317 (10%) were ex-smokers and 2,123 (67%) had never smoked. Only six (0.8%) of the 766 regular and occasional smokers were females. The rates of smoking among parents and siblings of smokers were higher than parents and siblings of non-smokers. Seventy-four percent of the smoking teachers admitted to smoking in the school premises. The teachers' attitudes about the health effects of smoking were statistically different between smokers and non-smokers. However, both smoking and non-smoking teachers had similar views regarding methods to control the smoking habit which included banning cigarette sales, putting a halt to the tobacco industry and banning cigarette advertisements.
A study of the smoking habits and attitudes toward smoking among 120 doctors at the Hospital Universiti Sains Malaysia was conducted between May to August 1991. Eighteen percent of the doctors were smokers, 13% ex-smokers and 69% had never smoked. All the smokers were male and all except one smoked only cigarettes. Three of the 32 female doctors were ex-smokers. Nineteen of the 21 smokers only smoked in areas where they could not be seen by the public. Most doctors (equally among smokers and non-smokers) had first-degree relatives (mostly males) who were smokers and 28% had relative with smoking-related disease. 81% non-smoking and 43% smoking doctors had advised healthy people to stop smoking. 92% non-smoking and 52% smoking doctors support the smoking-ban in the hospital. Seven of the 21 smokers had never attempted to quit smoking.
Malaysian infants would have to receive nine injections during the first few months of life in order to be protected against disease caused by hepatitis B (HBV), diphtheria, tetanus, pertussis and Haemophilus influenzae type b (Hib) if single HBV and Hib vaccines were used. We evaluated a combined DTPw-HBV/Hib vaccine administered at 1.5, 3 and 5 months after a birth dose of hepatitis B vaccine (HBV). One month after completion of the primary vaccination, 99% of subjects had seroprotective anti-HBV antibody levels, and at least 98% had seroprotective antibodies against diphtheria, tetanus, and Hib, and were seropositive for pertussis antibodies. The immune response to the combined vaccine was comparable to that induced by separate injections with DTPw, HBV and Hib vaccines. Overall, the DTPw-HBV/Hib vaccine was as well tolerated as separate administration of DTPw, HBV and Hib vaccines. The combined DTPw-HBV/Hib vaccine induces protection against five diseases as recommended in the Malaysian routine vaccination schedule. Use of the combined DTPw-HBV/Hib vaccine can reduce the required number of injections from nine to four in the first few months of life.
Cell disruption or lysis is a crucial step to obtain cellular components for various biological studies. We subjected different concentrations of Candida albicans to 5, 10, 15 and 20 cycles of disruption. The degree of cell lysis was observed using light microscopy and the yields obtained were measured and analysed. The optimum extraction with 1 x 10(10) yeast cells/ml was achieved after 5 cycles of disruption with 1.0 mm diameter glass beads at 5,000 rpm. Approximately 80% of the cells were lysed and the protein yield was 6,000 microg/ml. SDS-PAGE analysis revealed approximately 25 distinct protein bands with molecular weights ranging from 8 kDa to 220 kDa. We conclude that this mechanical disruption of fungal cells is a rapid, efficient and inexpensive technique for extracting whole cell proteins from yeast cells.
Globally, HIV/AIDS-related stigma and discriminatory attitudes deter the effectiveness of HIV prevention and care programs. This study investigated the general public's perceptions about HIV/AIDS-related stigma and discrimination towards people living with or affected by HIV/AIDS in order to understand the root of HIV/AIDS-related stigma and discriminatory attitudes. Study was carried out using qualitative focus group discussions (FGD). An interview guide with semi-structured questions was used. Participants were members of the public in Malaysia. Purposive sampling was adopted for recruitment of participants. A total 14 focus group discussions (n = 74) was carried out between March and July 2008. HIV/AIDS-related stigma and discrimination towards people living with HIV/AIDS (PLWHA) was profound. Key factors affecting discriminatory attitudes included high-risk taking behavior, individuals related to stigmatized identities, sources of HIV infection, stage of the disease, and relationship with an infected person. Other factors that influence attitudes toward PLWHA include ethnicity and urban-rural locality. Malay participants were less likely than other ethnic groups to perceive no stigmatization if their spouses were HIV positive. HIV/AIDS-related stigma and discrimination were stronger among participants in rural settings. The differences indicate attitudes toward PLWHA are influenced by cultural differences.
Two insular settlements (Kampung Pulau Ketam and Kampung Sungai Lima) were selected to study the population dynamics of Aedes aegypti and Aedes albopictus mosquitoes, vectors of dengue and chikungunya infections. Ovitrap surveillance was conducted between October 2007 and October 2008. There was an inverse negative association between ovitrap index and rainfall at the time of collection, probably because rainfall increased the number of available oviposition sites. Rainfall and ovitrap index were positively associates the 25th day after rainfall occurred. A minor, second peak was observed from the 38th to the 42nd day. The first peak was consistent with the minimum 18-day period between the hatching of eggs to the first oviposition. The second minor peak could be due to the second gonotrophic cycle of the female mosquitoes. Rainfall is an important environmental factor associated with Aedes breeding at the study sites.