Chikungunya is an acute febrile illness caused by chikungunya virus (CHIKV). In this study, the envelope E1 gene of CHIKV was cloned and expressed in a baculovirus system. The recombinant E1 protein with N-term 6-His residues protein was successfully expressed and purified as confirmed by SDS-PAGE and western blot analysis. The seroreactivity of the recombinant protein was evaluated in immunoassay for anti-CHIKV IgM and IgG antibodies. The recombinant antigen showed 69% sensitivity and 100% specificity for anti-CHIKV IgG by dot blot assay. Detection of anti-CHIKV IgM by dot assay showed 79% sensitivity and 100% specificity. No cross reactivity of the antigen was observed with anti-dengue virus serum samples. The results strongly support that the recombinant E1 protein has potential to be used as diagnostic antigen. The used of the antigen in a dot blot assay gives an advantage for laboratory detection without the need of any specialised equipment.
Monoclonal antibodies (mAbs) are unique and specific drug molecules targeting the treatment of various diseases such as arthritis, immune disorders, infectious diseases, and cancer etc. Different methods such as antibody coupled affinity chromatography, hydrophobic interaction chromatography, etc., can be applied to purify mAbs from various sources. This article provides a simple, cost effective, preparative native-polyacrylamide gel electrophoresis (n-PAGE)technique to purify mAbs expressed in H-192 cells (Hybridoma murine cell lines) against an antigen i.e. 17-alpha-hydroxyprogesterone (17-OHP), which further can have diagnostic application to detect Congenital Adrenal Hyperplasia (CAH). Furthermore, different parameters such as concentration and volume of the feedstock (medium containing antibodies), pore size of gel, height of resolving gel etc. were optimized to obtain the maximum purity and yield of mAbs.
MeSH terms: Adrenal Hyperplasia, Congenital; Animals; Antibodies, Monoclonal; Arthritis; Cell Line; Chromatography, Affinity; Hybridomas; Immune System Diseases; Neoplasms; 17-alpha-Hydroxyprogesterone; Mice; Hydrophobic and Hydrophilic Interactions; Native Polyacrylamide Gel Electrophoresis
Palm kernel cake (PKC), a by-product of the palm oil industry is limited in its use as a feed ingredient for poultry due to its high fibre and lignin content. The presence of these antinutritive components is the result of shells contaminating the by-product. The nutritive value of PKC has to be improved in order to increase its inclusion rate in poultry diet. In this study, PKC was subjected to a separation method using static cling and electrostatic separation to removethe shells present in PKC. Response surface methodology based on Box-Behnken design was used to optimize the separation method with moisture content (8 to 18%), particle size distribution (0.5 to 2.5 mm) and feed rate (20 to 200 g/min) as the independent variables evaluated. According to the regression coefficients and significance of the quadratic polynomial model, the optimum separation parameters were as follows: 13% PKC moisture content;
In response to the globalization of drug development, regulatory inspection of Good Clinical Practice (GCP) has recently been conducted not only by International Conference on Harmonisation (ICH) regions but also non-ICH regions. To promote the international implementation of GCP, consistent understanding and interpretation of its concept among regions are important. This article summarizes the background and past activities of the E6 Discussion Group, established under the Regulators Forum.
Investigation on in vivo effects of copper (Cu) on the ultrastructure of P. javanicus liver was
carried out using transmission electron microscopy (TEM). The addition of sublethal
concentration of 5 mg/L of Cu caused abnormalities on the bile canaliculi (BC) including
dilation and elongation compared to control and at lower concentrations of copper with a normal
round shape form. Findings from this study support an alternative histological assessment of the
effects of Cu concentration on P. javanicus liver.
MeSH terms: Bile Canaliculi; Copper; Dilatation; Liver; Microscopy, Electron, Transmission
Observations on the effects of copper on the liver proteome of Puntius javanicus based on the
one dimensional PAGE was carried out. The liver was dissected from each fish, which was
separately treated with different concentrations of copper sulfate ranging from 0.1 to 5.0 mg/L.
The livers were extracted and one dimensional PAGE was performed under nonreducing
(native) and reducing (SDS)-PAGE. Several bands were resolved in the native PAGE with
probable candidates for the effect of copper observed showing an increased in the expression
and downregulation strongly associated with increasing copper concentrations. This study
showed that high concentrations of copper significantly alters P. javanicus liver at the proteome
level, and preliminary screening based on one dimensional PAGE is considered rapid and
simple to assess the toxicity effect of copper before more advanced and extensive assesment
with a second dimensional PAGE is carried out.
The 3D structure of the insecticidal protein Cry1Ba4 produced by B. thuringiensis subsp.
Entomocidus HD-9 was determined using homology modelling. From the model built, we have
been able to identify the possible sites for structure modification by site-directed mutagenesis.
The mutation was introduced at the conserved region of -helix 7 by substituting the
hydrophobic motif that comprises alanine 216, leucine 217 and phenylalanine 218 with arginine.
Wild and mutant Cry1Ba4 genes were cloned into pET200/D-TOPO and expressed in the
expression host. The result suggests that mutant Cry1Ba4 protein was less toxic to the larvae
Plutella xylostella compared to the wild-type. In conclusion, alteration in the structure of
Domain I had left an impact on the toxicity of Cry1Ba4 against P. xylostella.
Chemical toxins and organic contaminants such as hydrocarbons and dyes are major global
contaminants with countless tones of those chemicals are created yearly with a significant
amount release to the environment. In this work we screen the ability of a molybdenum-reducing
bacterium isolated from contaminated soil to decolorize various azo and triphenyl methane dyes
independent of molybdenum reduction. Biochemical analysis resulted in a tentative identification
of the bacterium as Enterobacter sp. strain Zeid-6. The bacterium was able to decolorize the azo
dye Orange G. The bacterium reduces molybdate to Mo-blue optimally at pH between 5.5 and
8.0 and temperatures of between 30 and 37 oC. Other requirements include a phosphate
concentration of 5 mM and a molybdate concentration of 20 mM. The absorption spectrum of the
Mo-blue produced was similar to previous Mo-reducing bacterium, and closely resembles a
reduced phosphomolybdate. Molybdenum reduction was inhibited by copper, lead, mercury and
silver which showed 36.8, 16.9, 64.9 and 67.6% inhibition to Mo-reducing activity of
Enterobacter sp. strain Zeid-6, respectively. The resultant molybdenum blue spectrum closely
resembles the spectrum of molybdenum blue from the phosphate determination method. The
ability of this bacterium to detoxify molybdenum and decolorize azo dye makes this bacterium
an important tool for bioremediation.
The volume of contaminated rivers in Malaysia continues to keep rising through the years. The
cost of instrumental monitoring is uneconomical and prohibits schedule monitoring of
contaminants particularly heavy metals. In this work, a rapid enzyme assay utilizing the
molybdenum-reducing enzyme as an inhibitive assay, prepared in crude form from the
molybdenum-reducing bacterium Serratia sp. strain DRY5 has been developed for monitoring
the heavy metals mercury, silver, copper and chromium in contaminated waters in the Juru
Industrial Estate. The crude enzyme extract transformed soluble molybdenum
(phosphomolybdate) into a deep blue solution, which is inhibited by heavy metals such as
mercury, silver, copper and chromium. The IC50 and Limits of Detection (LOD) values for
mercury, copper, silver and cadmium were 0.245, 0.298, 0.367, 0.326, and 0.124, 0.086, 0.088
and 0.094 mg L-1, respectively. The assay is rapid, and can be carried out in less than 10 minutes.
In addition, the assay can be carried out at ambient temperature. The IC50 values for these heavy
metals are more sensitive than several established assays. Water samples from various locations
in the month of November from the Juru Industrial Estate (Penang) were tested for the presence
of heavy metals using the developed assay. Enzyme activity was nearly inhibited for water
samples from several locations. The presence of heavy metals was confirmed instrumentally
using Atomic Emission Spectrometry and a Flow Injection Mercury System. The assay is rapid
and simple and can be used as a first screening method for large scale monitoring of heavy
metals.
The pollution of heavy metals and toxic xenobiotics has become a central issue worldwide.
Bioremediation of these toxicants are being constantly carried out using novel microbes.
Molybdenum reduction to molybdenum blue is a detoxification process and mathematical
modelling of the reduction process can reveal important parameters such as specific reduction
rate, theoretical maximum reduction and whether reduction at high molybdenum concentration
affected the lag period of reduction. The used of linearization method through the use of natural
logarithm transformation, although popular, is inaccurate and can only give an approximate
value for the sole parameter measured; the specific growth rate. In this work, a variety of
models for such as logistic, Gompertz, Richards, Schnute, Baranyi-Roberts, Von Bertalanffy,
Buchanan three-phase and more recently Huang were utilized for the first time to obtain values
for the above parameters or constants. The modified Gompertz model was the best model in
modelling the Mo-blue production curve from Serratia marcescens strain DR.Y10 based on
statistical tests such as root-mean-square error (RMSE), adjusted coefficient of determination
(R2), bias factor (BF), accuracy factor (AF) and corrected AICc (Akaike Information Criterion).
Parameters obtained from the fitting exercise were maximum Mo-blue production rate (μm), lag
time (l) and maximal Mo-blue production (Ymax) of X (h-1), Y (h) and Z (nmole Mo-blue),
respectively. The application of primary population growth models in modelling the Moblue
production rate from this bacterium has become a successful undertaking. The model
may also be used in other heavy metals detoxification processes. The parameters
constants extracted from this work will be a substantial help for the future development
of further secondary models.
Water contamination by herbicides and chelating agents is increasing mainly due to the
increasing agricultural activities. Water contamination by these compounds has become a
concern due to their adverse effects to the environment and humans. Seven sampling sites of
water sources in Selangor and Johor were chosen for the study. Contamination level of
Mecoprop (MCCP), Nitrilotriacetic acid (NTA) and Ethylenediaminetetraacetic acid (EDTA) in
these water body areas was determined by using Gas Chromatography-Electron Capture
Detector (GC-ECD). Our results indicated that water samples of Sungai Melot in Selangor
showed the highest presence of EDTA. MCCP was detected at a high level at Sungai Sarang
Buaya, Johor while NTA showed similar level of concentration at three different sites, Ladang
10, Ladang Sayur and Mardi, Selangor.
The trend of global under-five child mortality rate showed a steady decrease at -3.14% per year
approaching the United Nations millennium development goals target (-4.0% reduction per year
from 1990-2015). This rate is usually inversely proportional to the nations GDP. However, the
progress in the recent years (2006-2012) in Malaysia has raised a disturbing concern with an
annual average increment of +0.81% per year. Thus, there is an urgent need to identify the
reasons of such risky ecosystem. By analyzing recorded data on river pollution from 2007-
2012, we found that river pollution has high correlation with the gross domestic products, not at
the same year, but for two years prior that indicates a lag time. We also found that under-five
child mortality has strong correlation with the river pollutions. The development-pollutionhealth
triangular cycle needs to be put in a balanced to ensure the nation prosperity and
sustainability of the nation.
Quinolines compounds are toxic pollutants. Their biodegradation by microbes represents a tool
for bioremediation. The growth of Klebsiella penumoniae on 2-methylquinoline shows typical
sigmoidal bacterial growth curves. Since there exists a variety of models for describing the
growth profile of microorganism such as logistic, Gompertz, Richards, Schnute, Baranyi-
Roberts, Von Bertalanffy, Buchanan three-phase and more recently Huang models, the growth
curves exhibit under such conditions would be an excellent study for finding the best model.
The Huang model was chosen as the best model based on statistical tests such as root-meansquare
error (RMSE), adjusted coefficient of determination (R2), bias factor (BF), accuracy
factor (AF) and corrected AICc (Akaike Information Criterion). Novel constants obtained from
the modelling exercise would be used for further secondary modelling.
Protein function depends greatly on its structure. Based on this principle, it is vital to study the
protein structure in order to understand its function. This study attempts to build the predicted
model of lipase gene in Rhodococcus sp. NAM81 using homology modelling method. The
predicted structure was then used to investigate the function of protein through several
bioinformatic tools. The DNA sequence of lipase gene was obtained from the Rhodococcus sp.
NAM81 genome scaffold. Blastx analysis showed 100% identity to the target enzyme andthe
appropriate template for homology modelling was determined using Blastp analysis. The 3D
protein structure was built using two homology modelling software, EsyPred3D and Swiss
Model Server. Both structures built obtained LGScore of greater than 4, which means they are
extremely good models according to ProQ validation criteria. Both structures also satisfied the
Ramachandran plot structure validation analysis. The predicted structures were 100% matched
with each other when superimposed with DaliLite pairwise. This shows that both structure
validation servers agreed on the same model. Structure analysis using ProFunc had found seven
motifs and active sites that indicate similar function of this protein with other known proteins.
Thus, this study has successfully produced a good 3D protein structure for the target enzyme.
Bioremediation is a new green economic approach in providing solutions for cleaning up
contaminated sites. Phytoremediation uses plants as a tool for remediation purposes. The usage
of plant species offers higher potential solution to remediate heavy metal contaminated sites.
This study aimed on screening potential plant species for phytoremediation of heavy metal
contaminated water. The potential of three aquatic macrophytes species (Eichorrnia crassipes,
Pistia stratiotes and Ipomoea aquatica) for chromium and nickel phytoremediations was tested.
The plants were exposed for 10 days under hydroponic conditions in heavy metal contaminated
water. E. crassipes showed the highest chromium and nickel concentrations in its biomass, 1.60
and 2.40 μg/L respectively. Meanwhile, P. stratiotes had chromium and nickel concentrations
detected at 0.89 and 0.081 μg/L, respectively; chromium and nickel concentrations of I.
aquatica detected were, 0.49 and 0.08 μg/L, respectively. The ability of these plants to
accumulate heavy metals and survived throughout the experiment demonstrates the potential of
these plants to remediate metal-enriched water. Among the three tested aquatic plants, E.
crassipes was proven to be the most suitable plant species that can phytoremediate heavy metal
contaminated water followed by P. stratiotes and I. aquatica.
Mass production of fish broodstock with high quality eggs requires the knowledge on the chemical composition and physiochemical properties of vitellogenin (Vtg) during ovulation. Vtg is an egg yolk precursor phospholipoglycoprotein, and has been analysed to evaluate the reproductive conditions and determine the spawning period in captive and wild fish. In this study, Vtg was induced in male H. nemurus through three intramuscular injections of 17-estradiol (E2). The Vtg was purified from the serum using gel filtration chromatography and the purified protein was reduced via SDS-PAGE. One major polypeptide corresponding to 130 kDa was observed. Vtg identification was done using peptide mass fingerprint (PMF) from the trypsin digestion of male H. nemurus Vtg induced with E2. The sequence homology of H. nemurus AYLAGAAADVLEVGVR matched the Vtg of other fish species when analysed using MALDI-TOF. Vtg was confirmed by MASCOT at 95% significant level. The potential protein that controls the reproductive process and oocyte development isolated from this study was discussed to understand the structure and function of Vtg.
This article define and discuss the issue of plagiarism by nursing students either in academic or clinical settings. It describes and explore the scenario of plagiarism among nursing students and implications for the qualification of the students. Currently, prevention is required to avoid negative culture in nursing education due to plagiarism.
This quantitative-oriented research was conducted to identify factors that contributed to errors in dispensing medication among nurses and to understand why nurses did not report their errors in dispensing. In this study a total of 284 U29 nurses participated in focusing on factors contributing to medication errors and failure to report the errors. In this study, analysis of the data collected was made in two sections; dispensing errors and failure to report the errors in giving medication. According to Evans et al. (2006) although nurses may not admit directly to such errors, they expressed their perceptions towards situations described in the questionnaire items as contributing to medication errors among nurses. Almost all in the sample of 284 chose not to report medication errors because they could not identify the cause of dispensing errors; other nurses perceived that the individual involved is not competent in performing the task. Other reasons include fear that the action will be exposed by the management, to avoid publicity from the media, and there is no difference in reporting or not reporting the medication errors. This study was done not only for exploring factors of medication errors; it also aspires to identify problems that arise in hospital services and in order to maintain the quality of health care. The management should consider the impact of medication errors and failure to report medication errors on the nursing profession and quality image of the hospital.
This article is a brief personal reflection on the ongoing development of professional education and practice in nursing. The reflection and its narratives are anecdotal and are based on the author's recent experience of teaching and working with fellow nursing lecturers in Malaysia in a nursing faculty within a local University. The author has almost 40 years career in nursing, nurse education and curriculum planning in the United Kingdom before her retirement. The study reflects on the key aspects of nursing and nurse education from her wealth of experiences while in the UK, and also highlights some positive and valuable insights gained from her brief experience in Malaysia. She also provides some reflections of the challenges that lie ahead, specifically, the emphasis on feelings and emotions in nursing, that provide the inspiration to continue with passion, dedication, and determination in our ongoing quest in the pursuit of nursing excellence for all the nurses, in practice and in education. This will hopefully and ultimately lead to a higher standard of nursing care for all patients, and continue the long journey towards the unique professional identity that we have been seeking, and that we can all proudly claim to be our own.
Family members' satisfaction has become an important measurement in determining the quality of care of patients in the critical care setting. The aims of the study were to determine the satisfaction level of family members, to determine the reliability of family satisfaction and its subscales, to identify mean of each question and their subscales. A cross-sectional study was conducted on 30 family members who accompanied patients during intensive care admission. The study was conducted in a critical care setting in
a high dependency unit (HDU) and cardiac intensive care unit (CCU), Hospital University Sains Malaysia (Hospital USM). The data was collected from October 2012 to January 2013. The Critical Care Family Satisfaction Survey (CCFSS) Malay version was used to measure family satisfaction in addition to the collection of demographic data. The statistical analysis used a descriptive approach. Ethical approval was obtained prior to data collection. Majority of the respondents were female: 23 (76.7%). Mean age was 35.67 years (SD=12.83) and the score of overall satisfaction level was 78.80 (SD=13.88). The Cronbach's alpha was 0.96. A majority of the respondents, 19 (63.3%) were satisfied with the care provided. This study showed that a majority of family members were slightly satisfied with the care that was provided in a critical
care setting at HUSM. It also showed that families play an important role in giving support and hope to patients.
Study site: HDU and CCU, Hospital Universiti Sains Malaysia (HUSM), Kelantan, Malaysia
MeSH terms: Adult; Critical Care; Cross-Sectional Studies; Demography; Female; Hospitals, University; Humans; Intensive Care Units; Malaysia; Personal Satisfaction; Surveys and Questionnaires; Universities; Reproducibility of Results