Mutation analysis of glycine decarboxylase, aminomethyltransferase and glycine cleavage system protein-H genes in 13 unrelated families with glycine encephalopathy.

Glycine encephalopathy (GCE) or nonketotic hyperglycinemia is an inborn error of glycine metabolism, inherited in an autosomal recessive manner due to a defect in any one of the four enzymes aminomethyltransferase (AMT), glycine decarboxylase (GLDC), glycine cleavage system protein-H (GCSH) and dehydrolipoamide dehydrogenase in the glycine cleavage system. This defect leads to glycine accumulation in body tissues, including the brain, and causes various neurological symptoms such as encephalopathy, hypotonia, apnea, intractable seizures and possible death. We screened 14 patients from 13 families with clinical and biochemical features suggestive of GCE for mutation in AMT, GLDC and GCSH genes by direct sequencing and genomic rearrangement of GLDC gene using a multiplex ligation-dependant probe amplification. We identified mutations in all 14 patients. Seven patients (50%) have biallelic mutations in GLDC gene, six patients (43%) have biallelic mutations in AMT gene and one patient (7%) has mutation identified in only one allele in GLDC gene. Majority of the mutations in GLDC and AMT were missense mutations and family specific. Interestingly, two mutations p.Arg265His in AMT gene and p.His651Arg in GLDC gene occurred in the Penan sub-population. No mutation was found in GCSH gene. We concluded that mutations in both GLDC and AMT genes are the main cause of GCE in Malaysian population.

Emergence of chikungunya seropositivity in healthy Malaysian adults residing in outbreak-free locations: chikungunya seroprevalence results from the Malaysian Cohort.

BACKGROUND: In 1998, Malaysia experienced its first chikungunya virus (CHIKV) outbreak in the suburban areas followed by another two in 2006 (rural areas) and 2008 (urban areas), respectively. Nevertheless, there is still a lack of documented data regarding the magnitude of CHIKV exposure in the Malaysian population. The aim of this study was to determine the extent of chikungunya virus infection in healthy Malaysian adults residing in outbreak-free locations. METHODS: A cross sectional study of chikungunya (CHIK) seroprevalence was carried out in 2009 amongst The Malaysian Cohort participants living in four states (Kuala Lumpur, Selangor, Pahang and Negeri Sembilan). A total of 945 participants were randomly identified for the study. Potential risk factors for CHIK infection were determined via questionnaires, and IgG antibodies against CHIK were detected by an enzyme-linked immunosorbent assay. Logistic regression identified risk factors associated with CHIK seropositivity, while geographical information system was used for visual and spatial analysis. RESULTS: From the 945 serum samples tested, 5.9% was positive for CHIK IgG. Being male, Malay, rural occupancy and Negeri Sembilan residency were identified as univariate predictors for CHIK seropositivity, while multivariate analysis identified being male and rural occupancy as risk factors. CONCLUSIONS: This study provided evidence that CHIK is slowly emerging in Malaysia. Although the current baseline seroprevalence is low in this country, increasing number of CHIK cases reported to the Malaysia Ministry of Health imply the possibility of CHIK virus becoming endemic in Malaysia.

Missense mutations in MLH1, MSH2, KRAS, and APC genes in colorectal cancer patients in Malaysia.

BACKGROUND: Colorectal cancer (CRC) is the third most common cancer worldwide with approximately 1 million cases diagnosed annually. In Malaysia, CRC is the second most common cancer in women and ranked first in men. The underlying cause of CRC remains unknown. AIMS: The aim of this study was to analyze the mutations in genes involved in CRC including MLH1, MSH2, KRAS, and APC genes. METHODS: A total of 76 patients were recruited. We used the polymerase chain reaction-denaturing high-performance liquid chromatography for the detection of mutations in the mismatch repair (MMR) and APC genes and the PCR single-strand conformation polymorphism for screening of the KRAS gene mutations. RESULTS: We identified 17 types of missense mutations in 38 out of 76 patients in our patients. Nine mutations were identified in the APC gene, five mutations were detected in the KRAS gene, and two mutations were identified in the MSH2 gene. Only one mutation was identified in MLH1. Out of these 17 mutations, eight mutations (47 %) were predicted to be pathogenic. Seven patients were identified with multiple mutations (3: MSH2 and KRAS, 1: KRAS and APC, 1: MLH1 and APC, 2: APC and APC). CONCLUSIONS: We have established the PCR-DHPLC and PCR-SSCP for screening of mutations in CRC patients. This study has given a snapshot of the spectrum of mutations in the four genes that were analyzed. Mutation screening in patients and their family members will help in the early detection of CRC and hence will reduce mortality due to CRC.

Evaluation of rational drug use based on World Health Organization prescribing indicators in a primary care center in Pamekasan East Java, Indonesia.

Background The Government of Indonesia has put in place many interventions for rationalizing drug use at all levels of the health services including in primary care centers (puskesmas). One of the programs for the rational use of drugs at the puskesmas is the monitoring and evaluation of drug use conducted by pharmacists. The purpose of this research was to evaluate the rationality of drug use in Pamekasan puskesmas that use World Health Organization (WHO) prescribing indicators and to find the difference in the percentage of rationality of drug use between puskesmas in Pamekasan. Methods This study reviewed official documents considering reporting of rational drug use. The documents were obtained from the district health office or from the public domain from 2014 to 2018. Data were then collated, extracted, and presented as frequencies. Results The percentage prescribed for antibiotics for acute respiratory infection (ARI) non-pneumonia was 47.27% and percentage prescribing antibiotic drugs in a non-specific diarrhea was 59.85%. The percentage prescribed for injection was 36.15%. The average number of drugs prescribed per treatment was 2.61. Conclusions The majority of WHO guidelines stated that prescribing indicators were not met by the puskesmas in Pamekasan, except for the parameters for the average number of drugs prescribed per consultation. This shows an alarming clarion call for the local healthcare stakeholders to improve such situations.

Evaluating current practices and policies in the use of injectable medicines for treating myalgia in a primary care center in Pamekasan, Indonesia.

Background Myalgia in patients can be associated with a large array of conditions, including injuries, infections and inflammations. Treatment for myalgia may include the use of oral, topical and injectable medicines (IM). However, the use of IM has been restricted by the World Health Organization due to the common hazards associated with inappropriate medicine use, risks of disease transmission and more expensive spending for using IM. Accordingly, the Ministry of Health of Indonesia (MoH) has limited the use at the level of ≤1% in every primary care center (Puskesmas) across the nation. The aim of this study was to report and evaluate rational drug use injection in myalgia in a Puskesmas in Pamekasan, Indonesia. Methods This study reviewed official documents, such as rational drug use report, medicine use databases and drug request report. The documents were obtained from the district health office or public domain from 2014 to 2018. Data were then collated, extracted and presented as frequencies. Results The average rate of use of IM for myalgia in Pamekasan was 36.15%. All 20 Puskesmas in Pamekasan conducted injection practice above the recommended level, thus reflecting a major gap between practice and policy implementation. This study implies that a substantial effort is needed to enforce the policy. Conclusions The use of IM for myalgia treatment in Pamekasan from 2014 to 2018 was significantly higher than the recommended level, thus reflecting a major problem in the practice. This is an alarming call for the local healthcare stakeholders to improve such a situation.

Exploration of barriers affecting job satisfaction among community pharmacists.

Background Understanding job satisfaction among community pharmacists is important as it may affect roles and performance. Several barriers in practice may affect job satisfaction. Objectives To explore barriers affecting job satisfaction among Indonesian community pharmacists. Methods This study reported data from the Nationwide Community Pharmacy Survey 2018. The results specific to participants in the East Java region were used for this study. Community pharmacists (507) participated in the survey between September 2018 and March 2019. The survey used questionnaire that asked 22 factors, which may act as barriers affecting job satisfaction. Data were analyzed using descriptive statistics. Results The majority of respondents were female (83.43%), and most respondents were aged 31-40 years (41.63%). Most respondents perceived nine barriers such as lack of time for interaction and lack of recognition that were dominant affecting job satisfaction. On the other hand, respondents were not in major agreement perceiving the remaining barriers as dominant affecting job satisfaction. More than half of the respondents disagreed that the remaining barriers were significant affecting job satisfaction. Conclusions This study identified some significant barriers affecting job satisfaction among community pharmacists. In general, barriers can arise from individual, management, and environmental issues suggesting a specific approach to reduce these barriers.

Mutational Profiles of F8 and F9 in a Cohort of Haemophilia A and Haemophilia B Patients in the Multi-ethnic Malaysian Population.

BACKGROUND: Haemophilia A (HA) and Haemophilia B (HB) are X-linked blood disorders that are caused by various mutations in the factor VIII (F8) and factor IX (F9) genes respectively. Identification of mutations is essential as some of the mutations are associated with the development of inhibitors. This study is the first comprehensive study of the F8 mutational profile in Malaysia. MATERIALS AND METHODS: We analysed 100 unrelated HA and 15 unrelated HB patients for genetic alterations in the F8 and F9 genes by using the long-range PCR, DNA sequencing, and the multiplex-ligation-dependent probe amplification assays. The prediction software was used to confirm the effects of these mutations on factor VIII and IX proteins. RESULTS: 44 (53%) of the severe HA patients were positive for F8 intron 22 inversion, and three (3.6%) were positive for intron one inversion. There were 22 novel mutations in F8, including missense (8), frameshift (9), splice site (3), large deletion (1) and nonsense (1) mutations. In HB patients, four novel mutations were identified including the splice site (1), small deletion (1), large deletion (1) and missense (1) mutation. DISCUSSION: The mutational spectrum of F8 in Malaysian patients is heterogeneous, with a slightly higher frequency of intron 22 inversion in these severe HA patients when compared to other Asian populations. Identification of these mutational profiles in F8 and F9 genes among Malaysian patients will provide a useful reference for the early detection and diagnosis of HA and HB in the Malaysian population.

Peripheral blood gene expression profile of atherosclerotic coronary artery disease in patients of different ethnicity in Malaysia.

The molecular basis of coronary artery disease (CAD) has been widely studied in the western world but there is no published work on the Malaysian population. This study looked at the global gene expression profiling of the peripheral blood of patients with CAD from the 3 main ethnic groups in Malaysia. Male subjects selected were based on angiographically confirmed CAD (≥50% stenosis) and normal control subjects (0% stenosis) with age range of 55.6±5.3 and 51.0±5.5 years, respectively. The global gene expression of 12 angiographically documented CAD patients and 11 matched control subjects were performed. The combined group samples identified 6 up regulated differential expression (DE) genes (GHRL, LTA, CBS, HP, ITGA2B, and OLR1) and 12 down regulated DE genes (IL18R1, ITGA2B, IL18RAP, HP, OLR1, SOD2 ITGB3, IL1B, MMP9, PLA2G7, UTS2, and CBS) to be involved in CAD at the fold change of 1.3 with fault discovery rate (FDR) of 1%. Three genes, MMP9, IL1B, and SOD2 were down regulated in all the 3 ethnic groups making them potential biomarker candidates for CAD across all three ethnicities. Further verification in a cohort study is needed.