Total concentrations and speciation of cadmium (Cd), copper (Cu), lead (Pb) and zinc (Zn) in surface sediment samples were correlated with the respective metal measured in the total soft tissue of the green-lipped mussel Perna viridis, collected from water off the west coast of Peninsular Malaysia. The aim of this study is to relate the possible differences in the accumulation patterns of the heavy metals in P. viridis to those in the surface sediment. The sequential extraction technique was employed to fractionate the sediment into 'freely leachable and exchangeable' (EFLE), 'acid-reducible,' 'oxidisable-organic' and 'resistant' fractions. The results showed that significant (P .05) was found between Zn in P viridis and all the sediment geochemical fractions of Zn and total Zn in the sediment. This indicated that Zn was possibly regulated from the soft tissue of P. viridis. The present results supported the use of P viridis as a suitable biomonitoring agent for Cd, Cu and Pb.
Malaysia is now a developing country and on her way towards being an industrialised one by the year 2020. Most of her industries and urban areas are located on the west coast of Peninsular Malaysia. In addition, the offshore area of the west coast is now one of the busiest shipping lanes in the world. These two phenomena make the intertidal and offshore areas of the west coast of Peninsular Malaysia interesting for scientific studies. Therefore, this study focused on both the offshore and intertidal sediments of the west coast of Peninsular Malaysia. Sampling for sediment samples were done from the northern to the southern ends of the peninsula and these sediment samples were analysed for Cu and Pb. It was found that total Cu concentrations ranged from 0.25 to 13.8 and 0.40 to 315 microg/g dry weight (dw) for offshore and intertidal sediments, respectively. For Pb, it ranged from 3.59 to 25.4 and 0.96 to 69.8 microg/g dw for the offshore and intertidal sediments, respectively. The ranges of Cu and Pb found from the west coast of Peninsular Malaysia were low in comparison to regional data. However, some intertidal areas were identified as receiving anthropogenic Cu and Pb. Geochemical studies revealed that the 'nonresistant' fraction for Pb contributed about 70.0% to 75.0% and 54.0% of the total Pb concentration in the offshore and intertidal sediments, respectively. As for Cu, the 'nonresistant' fraction contributed about 46.2% to 60.4% and 46.3% of the total Cu concentration in the offshore and intertidal sediments, respectively. The 'nonresistant' fraction contained mostly of anthropogenic metals besides natural origins. These 'nonresistant' percentages indicated that both the offshore and intertidal areas could have received anthropogenic-derived metals, which could be influenced by physico-chemical properties of the sediments. Although the present data indicated that contamination due to Cu and Pb in the west coast of Peninsular Malaysia especially in the offshore areas were not serious, regular biomonitoring studies along the west coast of Peninsular Malaysia are recommended.
Studies on toxicities and tolerances of cadmium (Cd), copper (Cu), lead (Pb) and zinc (Zn) in the brown alga Isochrysis galbana and in the green-lipped mussel Perna viridis were conducted by short-term bioassays using endpoints growth production and mortality, respectively. The 5-day EC(50) and 24-h LC(50) of these heavy metals were determined in the brown alga and mussel, respectively. The EC(50) values calculated for the alga were 0.74 mg/l for Cd, 0.91 mg/l for Cu, 1.40 mg/l for Pb and 0.60 mg/l for Zn. The LC(50) values for the mussels were 1.53 mg/l for Cd, 0.25 mg/l for Cu, 4.12 mg/l for Pb and 3.20 mg/l for Zn. These LC(50) values were within the concentration ranges as reported by other authors who used P. viridis as the test organism. Based on these EC(50) and LC(50) values, the alga was most sensitive to Zn, followed by Cd, Cu and Pb while the mussel was most sensitive to Cu, followed by Cd, Zn and Pb. Differences in the trophic levels, metal handling strategies, biology and ecology of the primary producer (brown alga) and the primary consumer (mussel) are believed to be the plausible causes for the different toxicities and tolerances of the metals studied.
It has been widely reported that heavy metal contamination in coastal waters can modify the allozyme profiles of marine organisms. Previous studies have recorded elevated metal concentrations in sediments and mussel tissues off Peninsular Malaysia. In the present study, horizontal starch gel electrophoresis was carried out to estimate the levels of allelic variation of the green-lipped mussel, Perna viridis, collected from one contaminated and three relatively uncontaminated sites off Peninsular Malaysia. Fourteen polymorphic loci were observed. In addition, the concentrations of cadmium, copper, lead, mercury and zinc were determined in the sediments and in the soft tissues of the mussels. Mussels from contaminated site, evidenced by high metal pollution indices (MPI) of the sediment and the mussel tissues, showed the highest percentage of polymorphic loci (78.6%), while those collected from the uncontaminated sites had lower MPI of the sediment and mussel tissue, and exhibited lower percentages of polymorphic loci (35.7-57.1%). The population from the contaminated site showed the highest excess of heterozygosity (0.289) when compared to that of the populations from the three uncontaminated sites (0.108-0.149). Allozyme frequencies at the phosphoglucomutase (PGM; E.C. 2.7.5.1) locus also differed between the contaminated and uncontaminated populations. Previous studies have shown that exposure to heavy metals can select or counter-select for particular alleles at this locus. The present results suggest that allozyme polymorphism in P. viridis is a potential biomonitoring tool for heavy metal contamination but further validation is required.
The rapid and effective method for the isolation of RNA from green microalga Ankistrodesmus convolutus based on homogenization in a simple CTAB buffer and selective precipitation of RNA with lithium chloride is developed. This procedure avoids the use of toxic chaotropic agents and phenol while high concentration of dithiothreitol is used to inhibit RNase activity and prevent oxidative cross-linking of nucleic acids by phenolics. The extraction procedure was able to produce high quality and intact RNA from A. convolutus. The yield of total RNA was 0.69-0.73 mg/g of fresh weight, with A(260)/A(280) ratio of 1.79-1.86. The obtained RNA was of sufficient quality and suitable for downstream application such as RT-PCR and cDNA library construction. The procedure may also have wider applicability for total RNA isolation from other green microalgae species.
Green microalga Ankistrodesmus convolutus Corda is a fast growing alga which produces appreciable amount of carotenoids and polyunsaturated fatty acids. To our knowledge, this is the first report on the construction of cDNA library and preliminary analysis of ESTs for this species. The titers of the primary and amplified cDNA libraries were 1.1×10(6) and 6.0×10(9) pfu/ml respectively. The percentage of recombinants was 97% in the primary library and a total of 337 out of 415 original cDNA clones selected randomly contained inserts ranging from 600 to 1,500 bps. A total of 201 individual ESTs with sizes ranging from 390 to 1,038 bps were then analyzed and the BLASTX score revealed that 35.8% of the sequences were classified as strong match, 38.3% as nominal and 25.9% as weak match. Among the ESTs with known putative function, 21.4% of them were found to be related to gene expression, 14.4% ESTs to photosynthesis, 10.9% ESTs to metabolism, 5.5% ESTs to miscellaneous, 2.0% to stress response, and the remaining 45.8% were classified as novel genes. Analysis of ESTs described in this paper can be an effective approach to isolate and characterize new genes from A. convolutus and thus the sequences obtained represented a significant contribution to the extensive database of sequences from green microalgae.
Over-expression of P-glycoprotein, encoded by the ABCB1 gene, is proposed to be involved in resistance to antiepileptic drugs in about 30% of patients with epilepsy. Here, we investigated the possible association between ABCB1 polymorphisms and sodium valproate (VPA) treatment in Malaysian epilepsy patients. Genotypes were assessed in 249 drug-resistant and 256 drug-responsive Malaysian patients for C1236T, G2677T/A, and C 5T polymorphisms in the ABCB1 gene. No genotypes, alleles, or haplotypes were associated with the response to VPA in either the overall group or Chinese, Indian, and Malay subgroups. Our data suggest that C1236T, G2677T/A, and C3435T polymorphisms in the ABCB1 gene do not contribute to the response to VPA in patients with epilepsy.
Haematococcus pluvialis (Flotow) is a unicellular green alga, which is considered to be the best astaxanthin-producing organism. Molecular markers are suitable tools for the purpose of finding out genetic variations in organisms; however there have been no studies conducted on ISSR or RAPD molecular markers for this organism. The DNA of 10 different strains of H. pluvialis (four strains from Iran, two strains from Finland, one strain from Switzerland and three strains from the USA) was extracted. A genetic similarity study was carried out using 14 ISSR and 12 RAPD primers. Moreover, the molecular weights of the bands produced ranged from 0.14 to 3.4 Kb. The PCA and dendrogram clustered the H. pluvialis strains into various groups according to their geographical origin. The lowest genetic similarity was between the Iran2 and USA2 strains (0.08) and the highest genetic similarity was between Finland1 and Finland2 (0.64). The maximum numbers of bands produced by the ISSR and RAPD primers were 35 and 6 bands, respectively. The results showed that ISSR and RAPD markers are useful for genetic diversity studies of Haematococcus as they showed geographical discrimination.
A phytochemical study of the bark of Alseodaphne perakensis has yielded three aporphine alkaloids: the new compound N-cyanomethylnorboldine (1), and the two known alkaloids N-methyllaurotetanine (2) and norboldine (3). The isolation was achieved by chromatographic techniques and the structural elucidation was performed via spectral methods, notably 1D- and 2D-NMR, UV, IR, and HRFABMS. The vasorelaxation activity of compound 1 has been studied.
An initial study on gene cloning and characterization of unicellular green microalga Ankistrodesmus convolutus was carried out to isolate and characterize the full-length cDNA of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) as a first step towards elucidating the structure of A. convolutus RbcS gene. The full-length of A. convolutus RbcS cDNA (AcRbcS) contained 28 bp of 5' untranslated region (UTR), 225 bp of 3' non-coding region, and an open reading frame of 165 amino acids consisting of a chloroplast transit peptide with 24 amino acids and a mature protein of 141 amino acids. The amino acid sequence has high identity to those of other green algae RbcS genes. The AcRbcS contained a few conserved domains including protein kinase C phosphorylation site, tyrosine kinase phosphorylation site and N-myristoylation sites. The AcRbcS was successfully expressed in Escherichia coli and a ~21 kDa of anticipated protein band was observed on SDS-PAGE. From the phylogenetic analysis of RbcS protein sequences, it was found that the RbcS of A. convolutus has closer genetic relationship with green microalgae species compared to those of green seaweed and green macroalgae species. Southern hybridization analysis revealed that the AcRbcS is a member of a small multigene family comprising of two to six members in A. convolutus genome. Under different illumination conditions, RT-PCR analysis showed that AcRbcS transcription was reduced in the dark, and drastically recovered in the light condition. Results presented in this paper established a good foundation for further study on the photosynthetic process of A. convolutus and other green algae species where little information is known on Rubisco small subunit.
Health economics, a special branch of science applying economic principles to the health delivery system, is a relatively young subdiscipline. The literature is scanty about teaching health economics in the medical and dental fields. Delivery methods of this topic vary from one university to another, with lectures, seminars, and independent learning reported as teaching/learning tools used for the topic. Ideally, debates should foster the development of logical reasoning and communication skills. Health economics in dentistry is taught under the community oral health module that constitutes part of an outcome-based dental curriculum in a private dental school in Kuala Lumpur, Malaysia. For this study, the students were divided into two groups: active participants (active debaters) and supporting participants (nonactive debaters). The debate style chosen for this activity was parliamentary style. Active and nonactive debaters' perceptions were evaluated before and after the activity through a structured questionnaire using a five-point rating scale addressing the topic and perceptions about debate as an educational tool. Cronbach's alpha coefficient was used as a measure of internal consistency for the questionnaire items. Among a total of eighty-two third-year dental students of two successive cohorts (thirty-eight students and forty-four students), seventy-three completed the questionnaire, yielding a response rate of 89 percent. Students' responses to the questionnaire were analyzed with the Kruskal-Wallis analysis of variance test. Results revealed that the students felt that their interest in debate, knowledge of the topic, and reinforcement of the previous knowledge had improved following participation in the debate. Within the limitations of this study, it can be concluded that debate was a useful tool in teaching health economics to dental students.
The continuous increase in number of people living with HIV/AIDS (PLWHA) represents a serious health and economic burden. HIV positive individuals with oral lesions have significantly lower oral health-related quality of life than HIV positive individuals without oral lesions. The objective of this study was to assess the knowledge, attitude and practices (KAP) within a cohort of HIV/AIDS positive patients towards HIV/AIDS associated oral lesions.
The human brain generates different oscillations at different frequencies during various consciousness levels. When these brain waves synchronize with exogenous rhythmic stimulation, the brain experiences strong, yet relaxing emotion that could be involved in the formation of memory. We investigated the character of rhythmic oscillatory dynamics by electroencephalography (EEG) of subjects listening to a short verse of the Holy Quran compared to resting and Arabic news listening. The mean power amplitudes of each frequency band for wavelet-based time-frequency analysis were obtained from 5000 ms of segmented EEG recordings during rest, news and Quran listening conditions. The time series analysis of power from each of three conditions in each frequency band from the grand averaged data was then subjected to autocorrelation study. The results showed significant cyclic overall trends of increasing and decreasing patterns of power in the low frequency brain wave oscillation of different head regions especially global, frontal and temporal sites. These results provided a basis for prediction of the periodicity of the power of the oscillatory brain dynamics of delta and robustly in theta regions which occurred during Quran listening. Despite several limitations, our data offered a plausible scientific basis to the emotional induction during Quran listening that mimics recognized as data from music listening studies. This offered a promising perspective for future studies in translational neurophysiological, cognitive and biofeedback on Quran listening to modify brain behaviour in health and disease.
Isolation of promoter sequences from known gene sequences is a tedious task in genome-related research. An efficient method of obtaining the promoter sequences is necessary in order to successfully use targeted promoters for genetic manipulations. Here, efficiency and usefulness of two PCR-based methods, namely: ligation-mediated PCR and thermal asymmetric interlaced (TAIL) PCR, for isolation of promoter sequences of the ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) gene from green microalgae Ankistrodesmus convolutus (A. convolutus) were evaluated. The results showed that the amplification efficiency of TAIL-PCR was higher than that of the ligation-mediated PCR method, i.e. the amplified promoter fragments of 1.2 and 0.8 kb in length or promoter sequences of 813 and 606 bp (after eliminating the unreadable sequences). The use of TAIL-PCR described here presents a low cost and efficient strategy for the isolation of promoter sequences of known genes, especially in GC-rich regions, and species with little or no available genome information such as A. convolutus.
The availability of highly active homologous promoters is critical in the development of a transformation system and improvement of the transformation efficiency. To facilitate transformation of green microalga Ankistrodesmus convolutus which is considered as a potential candidate for many biotechnological applications, a highly-expressed native promoter sequence of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (AcRbcS) has been used to drive the expression of β-glucuronidase (gusA) gene in this microalga. Besides the determination of the transcription start site by 5'-RACE, sequence analysis revealed that AcRbcS promoter contained consensus TATA-box and several putative cis-acting elements, including some representative light-regulatory elements (e.g., G-box, Sp1 motif and SORLIP2), which confer light responsiveness in plants, and several potential conserved motifs (e.g., CAGAC-motif, YCCYTGG-motifs and CACCACA-motif), which may be involved in light responsiveness of RbcS gene in green microalgae. Using AcRbcS promoter::gusA translational fusion, it was demonstrated that this promoter could function as a light-regulated promoter in transgenic A. convolutus, which suggested that the isolated AcRbcS promoter was a full and active promoter sequence that contained all cis-elements required for developmental and light-mediated control of gene expression, and this promoter can be used to drive the expression of heterologous genes in A. convolutus. This achievement therefore advances the development of A. convolutus as an alternative expression system for the production of recombinant proteins. This is the first report on development of gene manipulation system for unicellular green alga A. convolutus.
Student-generated videos provide an authentic learning experience for students, enhance motivation and engagement, improve communication skills, and improve collaborative learning skills. This article describes the development and implementation of a student-generated video activity as part of a knowledge, observation, simulation, and experience (KOSE) program at the School of Dentistry, International Medical University, Kuala Lumpur, Malaysia. It also reports the students' perceptions of an activity that introduced first-year dental students (n=44) to clinical scenarios involving patients and dental team aiming to improve professional behavior and communication skills. The learning activity was divided into three phases: preparatory phase, video production phase, and video-watching. Students were organized into five groups and were instructed to generate videos addressing given clinical scenarios. Following the activity, students' perceptions were assessed with a questionnaire. The results showed that 86 percent and 88 percent, respectively, of the students agreed that preparation of the activity enhanced their understanding of the role of dentists in provision of health care and the role of enhanced teamwork. In addition, 86 percent and 75 percent, respectively, agreed that the activity improved their communication and project management skills. Overall, the dental students perceived that the student-generated video activity was a positive experience and enabled them to play the major role in driving their learning process.