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1.  Induction of Apoptosis in MCF-7 Cells via Oxidative Stress Generation, Mitochondria-Dependent and Caspase-Independent Pathway by Ethyl Acetate Extract of Dillenia suffruticosa and Its Chemical Profile 
PLoS ONE  2015;10(6):e0127441.
Dillenia suffruticosa, which is locally known as Simpoh air, has been traditionally used to treat cancerous growth. The ethyl acetate extract of D. suffruticosa (EADs) has been shown to induce apoptosis in MCF-7 breast cancer cells in our previous study. The present study aimed to elucidate the molecular mechanisms involved in EADs-induced apoptosis and to identify the major compounds in the extract. EADs was found to promote oxidative stress in MCF-7 cells that led to cell death because the pre-treatment with antioxidants α-tocopherol and ascorbic acid significantly reduced the cytotoxicity of the extract (P<0.05). DCFH-DA assay revealed that treatment with EADs attenuated the generation of intracellular ROS. Apoptosis induced by EADs was not inhibited by the use of caspase-inhibitor Z-VAD-FMK, suggesting that the cell death is caspase-independent. The use of JC-1 dye reflected that EADs caused disruption in the mitochondrial membrane potential. The related molecular pathways involved in EADs-induced apoptosis were determined by GeXP multiplex system and Western blot analysis. EADs is postulated to induce cell cycle arrest that is p53- and p21-dependent based on the upregulated expression of p53 and p21 (P<0.05). The expression of Bax was upregulated with downregulation of Bcl-2 following treatment with EADs. The elevated Bax/Bcl-2 ratio and the depolarization of mitochondrial membrane potential suggest that EADs-induced apoptosis is mitochondria-dependent. The expression of oxidative stress-related AKT, p-AKT, ERK, and p-ERK was downregulated with upregulation of JNK and p-JNK. The data indicate that induction of oxidative-stress related apoptosis by EADs was mediated by inhibition of AKT and ERK, and activation of JNK. The isolation of compounds in EADs was carried out using column chromatography and elucidated using the nuclear resonance magnetic analysis producing a total of six compounds including 3-epimaslinic acid, kaempferol, kaempferide, protocatechuic acid, gallic acid and β-sitosterol-3-O-β-D-glucopyranoside. The cytotoxicity of the isolated compounds was determined using MTT assay. Gallic acid was found to be most cytotoxic against MCF-7 cell line compared to others, with IC50 of 36 ± 1.7 μg/mL (P<0.05). In summary, EADs generated oxidative stress, induced cell cycle arrest and apoptosis in MCF-7 cells by regulating numerous genes and proteins that are involved in the apoptotic signal transduction pathway. Therefore, EADs has the potential to be developed as an anti-cancer agent against breast cancer.
doi:10.1371/journal.pone.0127441
PMCID: PMC4457850  PMID: 26047480
2.  Prevalence of Oral Mucosal Lesions in Patients with Type 2 Diabetes Attending Hospital Universiti Sains Malaysia 
Objectives:
Diabetes mellitus is associated with a greater likelihood of developing certain oral mucosal disorders. This study was aimed at assessing the prevalence of oral mucosal lesions (OMLs) in patients with type 2 diabetes (DM2) and to investigate the association of such lesions with metabolic control of the disease.
Methods:
This cross-sectional study involved 391 patients with DM2 and 391 non-diabetic control subjects. Demographic information and data on the duration and type of diabetes, glycosylated hemoglobin (HbA1c) values, medical history, and current use of medication were obtained from medical records. Detailed oral examination was performed in accordance with international criteria.
Results:
The prevalence of OMLs was significantly higher among diabetic patients (45.5%) than among control subjects (38.4%) (P = 0.042). Patients with diabetes had a higher prevalence of geographic tongue (GT) (P = 0.017), denture stomatitis (P = 0.018), and angular cheilitis (P = 0.006) than controls. Overall, diabetic patients with poor metabolic control had a significantly higher prevalence of OMLs and xerostomia than patients with moderately and well-controlled disease (P < 0.05).
Conclusion:
The prevalence of OMLs was significantly higher in diabetic patients than in control subjects. Higher occurrence of OMLs was significantly associated with poor metabolic control.
PMCID: PMC3773351  PMID: 24043995
diabetes mellitus type 2; prevalence; mouth mucosa; oral pathology
3.  Detecting and Quantifying Forest Change: The Potential of Existing C- and X-Band Radar Datasets 
PLoS ONE  2015;10(6):e0131079.
This paper evaluates the opportunity provided by global interferometric radar datasets for monitoring deforestation, degradation and forest regrowth in tropical and semi-arid environments. The paper describes an easy to implement method for detecting forest spatial changes and estimating their magnitude. The datasets were acquired within space-borne high spatial resolutions radar missions at near-global scales thus being significant for monitoring systems developed under the United Framework Convention on Climate Change (UNFCCC). The approach presented in this paper was tested in two areas located in Indonesia and Australia. Forest change estimation was based on differences between a reference dataset acquired in February 2000 by the Shuttle Radar Topography Mission (SRTM) and TanDEM-X mission (TDM) datasets acquired in 2011 and 2013. The synergy between SRTM and TDM datasets allowed not only identifying changes in forest extent but also estimating their magnitude with respect to the reference through variations in forest height.
doi:10.1371/journal.pone.0131079
PMCID: PMC4482516  PMID: 26111047
4.  Identification of Subgingival Periodontal Pathogens and Association with the Severity of Periodontitis in Patients with Chronic Kidney Diseases: A Cross-Sectional Study 
BioMed Research International  2015;2015:370314.
Background. The aim of our study was to assess the subgingival profile of 9 periodontal pathogens, by means of real-time PCR, in a group of predialysis chronic kidney disease patients with and without periodontal disease and to identify the risk factors associated with periodontal disease in these patients. Material and Methods. This is a single centre cross-sectional cohort study performed on 70 CKD patients. Patients received a full-mouth periodontal examination and the following parameters were assessed: periodontal pocket depth (PPD), clinical attachment level, bleeding on probing, and plaque index; subgingival biofilm samples were collected from the deepest periodontal pocket of each quadrant and were pooled in one transporting unit. Clinical data were drawn from the medical file of the patients. Results. T. denticola (P = 0.001), T. forsythia (P < 0.001), and P. micros (P = 0.003) are significantly associated with periodontal disease in CKD subjects but in a multivariate model only age and T. forsythia remain independent risk factors for periodontal disease in patients with CKD. Conclusions. In our cohort, age and T. forsythia are independently associated with periodontitis in CKD patients. Within the limits of this study, CKD was not significantly associated with a particular subgingival periodontal pathogens profile in periodontitis patients.
doi:10.1155/2015/370314
PMCID: PMC4398940  PMID: 25922833
5.  The Prevalence of Parasitic Infestation of Small Ruminant Farms in Perak, Malaysia 
Helminthiasis due to strongyles such as Haemonchus contortus, coccidiosis caused by Eimeria sp. and blood parasite diseases such as theileriosis by Theileria sp. have been reported to cause severe morbidity and mortality annually in small ruminants in Malaysia. The aims of this study were to investigate the prevalence of helminthiasis, coccidiosis and theileriosis and to determine the packed cell volume (PCV) value of small ruminants in Perak, Malaysia. Blood and faecal samples were obtained from a total of 175 animals from 7 small ruminant farms in Kampar, Larut Matang and Selama, Kuala Kangsar and Manjung districts in Perak; the samples were examined for parasitic infestations from April to July 2011. The results of this study show that H. contortus was found in 152 (86.86%) animals, Eimeria sp. was found in 162 (92.57%) animals and the blood protozoa Theileria sp. was found in 25 (14.30%) animals. The PCV values of all of these animals were recorded between 7% and 44%. A total of 42 (24%) animals were anaemic, with a PCV of less than 21%. Continuous monitoring of small ruminant farms will provide important information for assisting farmers with managing the spread of parasitic infections and maintaining the productivity of animals.
PMCID: PMC4437319  PMID: 26019746
Helminthiasis; Coccidiosis; Theileriosis; Goat; Sheep; Helminthiasis; Coccidiosis; Theleriosis; Kambing; Kambing Biri-biri
6.  Phytochemicals from Mangifera pajang Kosterm and their biological activities 
Background
Mangifera pajang Kosterm is a plant species from the mango family (Anacardiaceae). The fruits are edible and have been reported to have high antioxidant content. However, the detailed phytochemical studies of the plant have not been reported previously. This study investigates the phytochemicals and biological activities of different parts of Mangifera pajang.
Methods
The plant samples were extracted with solvents of different polarity to obtain the crude extracts. The isolated compounds were characterized using spectroscopic methods. The extracts and isolated compounds were subjected to cytotoxicity tests using human breast cancer (MCF-7), human cervical cancer (HeLa) and human colon cancer (HT-29) cells. The free radical scavenging activity test was conducted using the DPPH assay. Antimicrobial activity tests were carried out by using the disc diffusion method.
Results
Phytochemical investigation on the kernel, stem bark and leaves of Mangifera pajang led to the isolation of methyl gallate (1), mixture of benzaldehyde (2) and benzyl alcohol (3), mangiferonic acid (4), 3β-hydroxy-cycloart-24-ene-26-oic acid (5), 3β,23-dihydroxy-cycloart-24-ene-26-oic acid (6), lupeol(7) lupenone(8), β-sitosterol(9), stigmasterol(10), trans-sobrerol(11) and quercitrin (12). Crude ethyl acetate and methanol extracts from the kernel indicated strong cytotoxic activity towards MCF-7 and HeLa cells with IC50 values of less than 10 μg/mL, while petroleum ether, chloroform and ethyl acetate extracts of the stem bark showed strong to moderate activity against MCF-7, HeLa and HT-29 cancer cell lines with IC50 values ranging from 5 to 30 μg/mL. As for the antimicrobial assays, only the ethyl acetate and methanol extracts from the kernel displayed some inhibition against the microbes in the antibacterial assays. The kernel extracts showed highest free radical scavenging activity with IC50 values of less than 10 μg/mL, while the ethyl acetate and methanol extracts of leaves displayed only weak activity in the DPPH assays.
Conclusions
Phytochemical investigations on various parts of Mangifera pajang have identified terpenoids and a flavonol derivative as major constituents. Bioassay studies have indicated that the crude extracts and isolated compounds have potential as naturally-derived anticancer and antimicrobial agents, besides possess high free radical scavenging activity.
doi:10.1186/s12906-015-0594-7
PMCID: PMC4391605  PMID: 25887035
Mangifera pajang; Bambangan; Phytochemicals; Cytotoxicity; DPPH; Antimicrobial
7.  Perceptions of students in different phases of medical education of the educational environment: Universiti Sultan Zainal Abidin 
Background
The Dundee Ready Education Environment Measure (DREEM) was planned and designed to quantify the educational environment precisely for medical schools and health-related professional schools. DREEM is now considered a valid and reliable tool, which is globally accepted for measuring the medical educational environment. The educational environment encountered by students has an impact on satisfaction with the course of study, perceived sense of well-being, aspirations, and academic achievement. In addition to being measurable, the educational environment can also be changed, thus enhancing the quality of medical education and the environment, and the medical education process. The objective of this study was to assess the educational environment of the Universiti Sultan Zainal Abidin (UniSZA) undergraduate medical program from the students’ perspective. The study expected to explore UniSZA medical students’ overall perceptions, perceptions of learning, teachers, atmosphere, academic self-perception, and social self-perception using the DREEM questionnaire.
Methods
A cross-sectional survey was conducted to study the perceptions of the students toward the educational environment of UniSZA as a new medical school, using the DREEM questionnaire. All medical students of UniSZA from Years I–V enrolled in the Bachelor of Medicine and Bachelor of Surgery programs were the target population (n=270). Therefore, the universal sampling technique was used. The data were analyzed using the SPSS 20 software. This study obtained ethical clearance from the Faculty of Medicine and Health Sciences, UniSZA.
Results
A total of 195 out of 270 students responded. Respondents included 31% males and 69% females. The overall DREEM scores were significantly higher (P<0.001) for females than males.
Conclusion
The medical students at UniSZA showed a positive perception of their educational environment. The new medical faculty, established for only a few years, has achieved an above-average, conducive educational environment for students. Most of the students showed a positive perception for the entire five domains tested in the DREEM survey. Females were consistently satisfied with UniSZA’s educational environment, and self-perception was high, as compared to male undergraduates.
Video abstract
doi:10.2147/AMEP.S78838
PMCID: PMC4378299  PMID: 25848333
perceptions; medical students; educational environment; DREEM; UniSZA
8.  Assessment of Routine Immunization Coverage in Nyala Locality, Reasons behind Incomplete Immunization in South Darfur State, Sudan 
Little is known about the coverage of routine immunization service in South Darfur state, Sudan. Therefore, this study was conducted to determine the vaccination rate and barriers for vaccination. A cross-sectional community-based study was undertaken in Nyala locality, south Darfur, Sudan, including urban, rural and Internal Displaced Peoples (IDPs) population in proportional representation. Survey data were collected by a questionnaire which was applied face to face to parents of 213 children 12-23 months. The collected data was then analyzed with SPSS software package. Results showed that vaccination coverage as revealed by showed vaccination card alone was 63.4% while it was increased to 82.2% when both history and cards were used. Some (5.6%) of children were completely non-vaccinated. The factors contributing to the low vaccination coverage were found to be knowledge problems of mothers (51%), access problems (15%) and attitude problems (34%). Children whose mother attended antenatal care and those from urban areas were more likely to complete their immunization schedule. In conclusion, the vaccination coverage in the studied area was low compared to the national coverage. Efforts to increase vaccination converge and completion of the scheduled plan should focus on addressing concerns of caregivers particularly side effects and strengthening the Expanded Programmer on Immunization services in rural areas.
PMCID: PMC4340504  PMID: 25729558
Expanded programme on immunization; routine immunization; vaccination coverage
9.  Low Serum High Density Lipoprotein Cholesterol Concentration is an Independent Predictor for Enhanced Inflammation and Endothelial Activation 
PLoS ONE  2015;10(1):e0116867.
Background
Inflammation, endothelial activation and oxidative stress have been established as key events in the initiation and progression of atherosclerosis. High-density lipoprotein cholesterol (HDL-c) is protective against atherosclerosis and coronary heart disease, but its association with inflammation, endothelial activation and oxidative stress is not well established.
Objectives
(1) To compare the concentrations of biomarkers of inflammation, endothelial activation and oxidative stress in subjects with low HDL-c compared to normal HDL-c; (2) To examine the association and correlation between HDL-c and these biomarkers and (3) To determine whether HDL-c is an independent predictor of these biomarkers.
Methods
422 subjects (mean age±SD = 43.2±11.9years) of whom 207 had low HDL-c concentrations (HDL-c <1.0mmol/L and <1.3mmol/L for males and females respectively) and 215 normal controls (HDL-c ≥1.0 and ≥1.3mmol/L for males and females respectively) were recruited in this study. The groups were matched for age, gender, ethnicity, smoking status, diabetes mellitus and hypertension. Fasting blood samples were collected for analysis of biomarkers of inflammation [high-sensitivity C-reactive protein (hsCRP) and Interleukin-6 (IL-6)], endothelial activation [soluble Vascular Cell Adhesion Molecule-1 (sVCAM-1), soluble Intercellular Adhesion Molecule-1 (sICAM-1) and E-selectin)] and oxidative stress [F2-Isoprostanes, oxidized Low Density Lipoprotein (ox-LDL) and Malondialdehyde (MDA)].
Results
Subjects with low HDL-c had greater concentrations of inflammation, endothelial activation and oxidative stress biomarkers compared to controls. There were negative correlations between HDL-c concentration and biomarkers of inflammation (IL-6, p = 0.02), endothelial activation (sVCAM-1 and E-selectin, p = 0.029 and 0.002, respectively), and oxidative stress (MDA and F2-isoprostane, p = 0.036 and <0.0001, respectively). Multiple linear regression analysis showed HDL-c as an independent predictor of IL-6 (p = 0.02) and sVCAM-1 (p<0.03) after correcting for various confounding factors.
Conclusion
Low serum HDL-c concentration is strongly correlated with enhanced status of inflammation, endothelial activation and oxidative stress. It is also an independent predictor for enhanced inflammation and endothelial activation, which are pivotal in the pathogenesis of atherosclerosis and atherosclerosis-related complications.
doi:10.1371/journal.pone.0116867
PMCID: PMC4304817  PMID: 25614985
10.  CT and MR Imaging in a Large Series of Patients with Craniofacial Fibrous Dysplasia 
Polish Journal of Radiology  2015;80:232-240.
Summary
Background
In this retrospective review of patients with craniofacial fibrous dysplasia (FD), the clinical and radiological findings of CT and MR scan were analyzed.
Material/Methods
The study material included 32 patients, at 9 to 68 years of age that were directed for differential diagnostics of several disorders in the head. We recorded CT and MRI data related to the lesion number, location, sidedness, appearance, and sex of the cases with craniofacial FD.
Results
Of 32 patients involved in this study, 17 had monostotic and 15 had polyostotic involvement pattern. Bones most commonly involved by monostotic involvement in females were, in descending order, mandibular, maxillary, and sphenoid bones, while the sphenoid bone was involved the most in males. Leontiasis ossea was observed in 2 patients. Sclerotic and mixed lesion types were more common in both females and males. In T1- and T2-weighted MRI sequences, hypointensity was more common compared to hyperintensity or heterogeneous intensity. The type of enhancement of lesions was found similar after contrast medium administration.
Conclusions
In the presence of craniofacial FD during CT or MRI imaging of the head, a detailed description of FD lesions may provide an important clinical benefit by increasing radiological experience during the diagnostics of this rare disorder.
doi:10.12659/PJR.893425
PMCID: PMC4424911  PMID: 26000068
Craniofacial Abnormalities; Fibrous Dysplasia of Bone; Magnetic Resonance Imaging; Tomography Scanners, X-Ray Computed
11.  Detection of airflow limitation using a handheld spirometer in a primary care setting 
Respirology (Carlton, Vic.)  2014;19(5):689-693.
Background and objective
Early diagnosis of chronic obstructive pulmonary disease (COPD) in primary care settings is difficult to achieve chiefly due to lack of availability of spirometry. This study estimated the prevalence of airflow limitation among chronic smokers using a handheld spirometer in this setting.
Methods
This is a cross-sectional study performed on consecutive patients who were ≥40 years old with ≥10 pack-years smoking history. Face-to-face interviews were carried out to obtain demographic data and relevant information. Handheld spirometry was performed according to a standard protocol using the COPd-6 device (Model 4000, Vitalograph, Ennis, Ireland) in addition to standard spirometry. Airflow limitation was defined as ratio of forced expiratory volume in 1 s (FEV1)/forced expiratory volume in 6 s <0.75 (COPd-6) or FEV1/forced vital capacity <0.7. Multiple logistic regression analyses were used to determine predictors of airflow limitation.
Results
A total of 416 patients were recruited with mean age of 53 years old. The prevalence of airflow limitation was 10.6% (n = 44) with COPd-6 versus 6% as gauged using standard spirometry. Risk factors for airflow limitation were age >65 years (odds ratio (OR) 3.732 95% confidence interval (CI): 1.100–1.280), a history of ‘bad health’ (OR 2.524, 95% CI: 1.037–6.142) and low to normal body mass index (OR 2.914, 95% CI: 1.191–7.190).
Conclusions
In a primary care setting, handheld spirometry (COPd-6) found a prevalence of airflow limitation of ∼10% in smokers. Patients were older, not overweight and had an ill-defined history of health problems.
SUMMARY AT A GLANCE
Prevalence of COPD is unknown in Malaysia. The prevalence of COPD using a handheld spirometer (COPd-6TM) was 10.6% versus 6% as gauged using standard spirometry. Predictors of COPD were older age, lower BMI and a history of ‘bad health’. Case-finding for COPD should be targeted in this special population.
doi:10.1111/resp.12291
PMCID: PMC4230390  PMID: 24708063
chronic obstructive pulmonary disease; Malaysia; prevalence; primary care; smoke
12.  A new isometric quadriceps-strengthening exercise using EMG-biofeedback 
A new isometric contraction quadriceps-strengthening exercise was developed to restore the quadriceps strength lost after knee surgery more rapidly. This study evaluated the results of this new method. Patients were taught to perform the isometric quadriceps-strengthening exercise in the unaffected knee in the supine position, and then they performed it in the affected knee. First, patients were taught the classical isometric quadriceps-strengthening exercise, and then they were taught our new alternative method: “pull the patella superiorly tightly and hold the leg in the same position for 10 seconds”. Afterward, the quadriceps contraction was evaluated using a non-invasive Myomed 932 EMG-biofeedback device (Enraf-Nonius, The Netherlands) with gel-containing 48 mm electrodes (Türklab, The Turkey) placed on both knees. The isometric quadriceps-strengthening exercise performed using our new method had stronger contraction than the classical method (P < 0.01). The new method involving pulling the patella superiorly appears to be a better choice, which can be applied easily, leading to better patient compliance and greater quadriceps force after arthroscopic and other knee surgeries.
PMCID: PMC4211772  PMID: 25356122
EMG biofeedback; quadriceps; strengthening; exercise
13.  Multiplex amplification refractory mutation system (MARMS) for the detection of β-globin gene mutations among the transfusion-dependent β-thalassemia Malay patients in Kelantan, Northeast of Peninsular Malaysia 
The aim of this study was to adapt MARMS with some modifications to detect beta mutation in our cohort of thalassemia patients. We focused only on transfusion-dependent thalassemia Malay patients, the predominant ethnic group (95%) in the Kelantanese population. Eight mutations were identified in 46 out of 48 (95.83%) beta thalassemia alleles. Most of the patients (54.2%) were compound heterozygous with co-inheritance Cd 26 (G>A). The frequencies of spectrum beta chain mutation among these patients are presented in Table 2. Among the transfusion dependent beta thalassemia Malay patients studied, 26 patients were found to be compound heterozygous and the main alleles were Cd 26 (G>A). Compound heterozygous mutation of Cd 26 (G>A) and IVS 1-5 (G>C) were 12 (46.2%), Cd 26 (G>A) and Cd 41/42 (TTCT) were 9 (34.6%), Cd 26 (G>A) and IVS 1-1 (G>C) were 2 (7.7%) respectively. Meanwhile the minority were made of a single compound heterozygous of Cd 26 (G>A) and Cd 71/72, Cd 26 (>A) and Cd 17 (A>T), Cd 26 (G>A) and -28 (G>A) respectively. Twenty out of forty six patients were shown to have homozygous of IVS 1-5 (G>C) were 2 (10.0%), Cd 26 (G>A) were 15 (75.0%), Cd 19 (A>G) were 1 (5.0%), and IVS 1-1 (G>T) were 2 (10.0%). The beta chain mutations among the Kelantanese Malays followed closely the distribution of beta chain mutations among the Thais and the Malays of the Southern Thailand. The G-C transition at position 5 of the IVS 1-5 mutation was predominant among the Malay patients. In conclusion, this method has successfully identified the mutation spectrum in our cohort of transfusion-dependent beta thalassemia patients, and this method is equally effective in screening for mutation among thalassemia patients.
PMCID: PMC4165115  PMID: 25232503
MARMS-PCR; β-globin gene; thalassemia; Malay; mutation
14.  A simple method to improve adenoma detection rate during colonoscopy: Altering patient position 
BACKGROUND:
Colonoscopy is currently considered to be the gold standard method for detecting and removing adenomatous polyps. However, tandem colonoscopy studies reveal a pooled polyp miss rate of 22%.
OBJECTIVE:
A prospective randomized trial was conducted to assess whether alteration of patient position during colonoscopy withdrawal increases the adenoma detection rate (ADR).
METHOD:
The study group included 120 patients who presented for elective colonoscopic examination. After reaching the cecum, patients were randomly assigned in a 1:1 ratio to examination in either the left lateral position or other positions (left lateral position for the cecum, ascending colon and hepatic flexure; supine for transverse colon; and supine and right lateral position for splenic flexure, descending and sigmoid colon) first. Examination of the colon was performed segment by segment. The size, morphology and location of all polyps were recorded. Polyps were removed immediately after examination of a colon segment when all positions were completed. ADR and polyp detection rates (PDR) were calculated.
RESULTS:
A total of 102 patients completed the study. Examination in the left lateral position revealed 66 polyps in 31 patients (PDR 30.3%) and 42 adenomas in 24 patients (ADR 23.5%). PDR increased to 43.1% (81 polyps in 44 patients) and the ADR to 33.3% (53 adenomas in 34 patients) after the colon was examined in the additional positions (P<0.001 and P=0.002, respectively). The increase in the number of adenomas detected was statistically significant in the transverse and sigmoid colon. The addition of position changes led to a 9.8% increase in the ADR in the transverse colon, splenic flexure, and descending and sigmoid colon. The frequency of surveillance interval was shortened in nine (8.8%) patients after examination of the colon in dynamic positions.
CONCLUSION:
Alteration of patient position during colonoscopy withdrawal is a simple and effective method to improve ADR.
PMCID: PMC3956008  PMID: 24078934
Adenoma detection rate; Adenomatous polyp; Colon; Colonoscopy
15.  Inhibition of Corneal Neovascularization by Topical and Subconjunctival Tigecycline 
Journal of Ophthalmology  2014;2014:452685.
Objective. To investigate the effects of topical and subconjunctival tigecycline on the prevention of corneal neovascularization. Materials and Methods. Following chemical burn, thirty-two rats were treated daily with topical instillation of 1 mg/mL tigecycline (group 1) or subconjunctival instillation of 1 mg/mL tigecycline (group 3) for 7 days. Control rats received topical (group 2) or subconjunctival (group 4) 0.9% saline. Digital photographs of the cornea were taken on the eighth day after treatment and analyzed to determine the percentage area of the cornea covered by neovascularization. Corneal sections were analyzed histopathologically. Results. The median percentages of corneal neovascularization in groups 1 and 3 were 48% (95% confidence interval (CI), 44.2–55.8%) and 33.5% (95% CI, 26.6–39.2%), respectively. The median percentages of corneal neovascularization of groups 1 and 3 were significantly lower than that of the control group (P = 0.03 and P < 0.001, resp.). Histologic examination of samples from groups 1 and 3 showed lower vascularity than that of control groups. Conclusion. Topical and subconjunctival administration of tigecycline seems to be showing promising therapeutic effects on the prevention of corneal neovascularization. Furthermore, subconjunctival administration of tigecycline is more potent than topical administration in the inhibition of corneal neovascularization.
doi:10.1155/2014/452685
PMCID: PMC4150514  PMID: 25197558
16.  Polycomb repressive complex 2 contributes to DNA double-strand break repair 
Cell Cycle  2013;12(16):2675-2683.
Polycomb protein histone methyltransferase, enhancer of Zeste homolog 2 (EZH2), is frequently overexpressed in human malignancy and is implicated in cancer cell proliferation and invasion. However, it is largely unknown whether EZH2 has a role in modulating the DNA damage response. Here, we show that polycomb repressive complex 2 (PRC2) is recruited to sites of DNA damage. This recruitment is independent of histone 2A variant X (H2AX) and the PI-3-related kinases ATM and DNA-PKcs. We establish that PARP activity is required for retaining PRC2 at sites of DNA damage. Furthermore, depletion of EZH2 in cells decreases the efficiency of DSB repair and increases sensitivity of cells to gamma-irradiation. These data unravel a crucial role of PRC2 in determining cancer cellular sensitivity following DNA damage and suggest that therapeutic targeting of EZH2 activity might serve as a strategy for improving conventional chemotherapy in a given malignancy.
doi:10.4161/cc.25795
PMCID: PMC3865057  PMID: 23907130
PARP; DNA damage; polycomb group proteins; chromatin; epigenetics
17.  Parasitism Performance and Fitness of Cotesia vestalis (Hymenoptera: Braconidae) Infected with Nosema sp. (Microsporidia: Nosematidae): Implications in Integrated Pest Management Strategy 
PLoS ONE  2014;9(6):e100671.
The diamondback moth (DBM) Plutella xylostella (L.) has traditionally been managed using synthetic insecticides. However, the increasing resistance of DBM to insecticides offers an impetus to practice integrated pest management (IPM) strategies by exploiting its natural enemies such as pathogens, parasitoids, and predators. Nevertheless, the interactions between pathogens and parasitoids and/or predators might affect the effectiveness of the parasitoids in regulating the host population. Thus, the parasitism rate of Nosema-infected DBM by Cotesia vestalis (Haliday) (Hym., Braconidae) can be negatively influenced by such interactions. In this study, we investigated the effects of Nosema infection in DBM on the parasitism performance of C. vestalis. The results of no-choice test showed that C. vestalis had a higher parasitism rate on non-infected host larvae than on Nosema-treated host larvae. The C. vestalis individuals that emerged from Nosema-infected DBM (F1) and their progeny (F2) had smaller pupae, a decreased rate of emergence, lowered fecundity, and a prolonged development period compared to those of the control group. DBM infection by Nosema sp. also negatively affected the morphometrics of C. vestalis. The eggs of female C. vestalis that developed in Nosema-infected DBM were larger than those of females that developed in non-infected DBM. These detrimental effects on the F1 and F2 generations of C. vestalis might severely impact the effectiveness of combining pathogens and parasitoids as parts of an IPM strategy for DBM control.
doi:10.1371/journal.pone.0100671
PMCID: PMC4072679  PMID: 24968125
18.  Induction of cell cycle arrest and apoptosis in caspase-3 deficient MCF-7 cells by Dillenia suffruticosa root extract via multiple signalling pathways 
Background
Dillenia suffruticosa root dichloromethane extract (DCM-DS) has been reported to exhibit strong cytotoxicity towards breast cancer cells. The present study was designed to investigate the cell cycle profile, mode of cell death and signalling pathways of DCM-DS-treated human caspase-3 deficient MCF-7 breast cancer cells.
Methods
Dillenia suffruticosa root was extracted by sequential solvent extraction. The anti-proliferative activity of DCM-DS was determined by using MTT assay. The mode of cell death was evaluated by using inverted light microscope and Annexin-V/PI-flow cytometry analysis. Cell cycle analysis and measurement of intracellular reactive oxygen species (ROS) were performed by using flow cytometry. MCF-7 cells were co-treated with antioxidants α-tocopherol and ascorbic acid to evaluate whether the cell death was mainly due to oxidative stress. GeXP-based multiplex system was employed to investigate the expression of apoptotic, growth and survival genes in MCF-7 cells. Western blot analysis was performed to confirm the expression of the genes.
Results
DCM-DS was cytotoxic to the MCF-7 cells in a time-and dose-dependent manner. The IC50 values of DCM-DS at 24, 48 and 72 hours were 20.3 ± 2.8, 17.8 ± 1.5 and 15.5 ± 0.5 μg/mL, respectively. Cell cycle analysis revealed that DCM-DS induced G0/G1 and G2/M phase cell cycle arrest in MCF-7 cells at low concentration (12.5 and 25 μg/mL) and high concentration (50 μg/mL), respectively. Although Annexin-V/PI-flow cytometry analysis has confirmed that DCM-DS induced apoptosis in MCF-7 cells, the distinct characteristics of apoptosis such as membrane blebbing, chromatin condensation, nuclear fragmentation and formation of apoptotic bodies were not observed under microscope. DCM-DS induced formation of ROS in MCF-7 cells. Nevertheless, co-treatment with antioxidants did not attenuate the cell death at low concentration of DCM-DS. The pro-apoptotic gene JNK was up-regulated whereby anti-apoptotic genes AKT1 and ERK1/2 were down-regulated in a dose-dependent manner. Western blot analysis has confirmed that DCM-DS significantly up-regulated the expression of pro-apoptotic JNK1, pJNK and down-regulated anti-apoptotic AKT1, ERK1 in MCF-7 cells.
Conclusion
DCM-DS induced cell cycle arrest and apoptosis in MCF-7 cells via multiple signalling pathways. It shows the potential of DCM-DS to be developed to target the cancer cells with mutant caspase-3.
doi:10.1186/1472-6882-14-197
PMCID: PMC4096536  PMID: 24947113
Dillenia suffruticosa; Dichloromethane extract; Cell cycle; Apoptosis; Oxidative stress
19.  A Giant Juvenile Nasopharyngeal Angiofibroma 
The Journal of craniofacial surgery  2013;24(3):e207-e209.
Juvenile nasopharyngeal angiofibroma (JNA) are locally growing highly vascular tumours. They are treated primarily by surgical excision ranging from open approach to endoscopic approach. We presented a 20-year-old male with a giant nasopharyngeal juvenile angiofibroma obliterating the pterygopalatine fossa bilaterally, invasing the sphenoid bone and extending to the left nasal passage. His complaints were epistaxis and nasal obstruction. After embolization, the patient was treated surgically with endoscopic approach and discharged as cured without any complication.
doi:10.1097/SCS.0b013e318268cf2e
PMCID: PMC3668308  PMID: 23714961
Juvenile nasopharyngeal angiofibroma; Endoscopic surgery; Bipolar cautery
20.  Indications of Brain Computed Tomography Scan in Children Younger Than 3 Years of Age with Minor Head Trauma 
Objective. To investigate the indications to receive brain computed tomography (CT) scan and to define the pathological findings in children younger than three years of age with minor head trauma in emergency departments. Methods. In this study, hospital case notes of 1350 children attending the emergency department of Bitlis State Hospital between January 2011 and June 2013 were retrospectively reviewed. 508 children under 3 years of age with minor head trauma were included in this study. We also asked 37 physicians about the indications for requiring CT in these children. Results. This study included 508 children, 233 (45,9%) of whom were female and 275 were male. In 476 (93,7%) children, the brain CT was completely normal. 89,2% of physicians asked in the emergency department during that time interval reported that they requested CT scan to protect themselves against malpractice litigation. Conclusion. In infants and children with minor head trauma, most CT scans were unnecessary and the fear of malpractice litigation of physicians was the most common reason for requesting a CT.
doi:10.1155/2014/248967
PMCID: PMC3958781  PMID: 24724031
21.  The Effects of Babesiosis on Oxidative Stress and DNA Damage in Anatolian Black Goats Naturally Infected with Babesia ovis 
Background
A reactive oxygen and nitrogen intermediate produced during an inflammatory response is the important part of host-defense strategies of organisms to kill the parasite. However, it is not well known whether these intermediates cause DNA damage and oxidative stress in goats infected with Babesia ovis. The purpose of this study was to clarify the effects of babesiosis on basal levels of DNA damage and oxidative status of goats naturally infected with B.ovis.
Methods
DNA damage and antioxidant parameters were determined in B. ovis infected goats. Ten infected Anatolian Black Goats with B. ovis diagnosed via clinical signs and microscopic findings and ten healthy were used in the study.
Results
The Babesia infection increased the levels of DNA damage, malondialdehyde (MDA), protein carbonyl content (PCO) and plasma concentration of nitric oxide metabolites (NOx), and decreased total antioxidant activities (AOA) and reduced glutathione (GSH). A significant positive correlation between DNA damage, MDA, PCO, and NOx concentrations was found in the infected goats. DNA damage showed a negative association with AOA and GSH concentrations in the infected goats.
Conclusion
The Babesia infection increases oxidative stress markers and DNA damage and decreases AOA in goats. These results suggest that the increases in the production of free radicals due to Babesia infection not only contribute to host-defense strategies of organisms to kill the parasite but also induce oxidative damage in other cells.
PMCID: PMC4289885  PMID: 25642264
Babesia ovis; oxidative stress; DNA damage; Antioxidant activity
22.  Induction of apoptosis through oxidative stress-related pathways in MCF-7, human breast cancer cells, by ethyl acetate extract of Dillenia suffruticosa 
Background
Breast cancer is one of the most dreading types of cancer among women. Herbal medicine has becoming a potential source of treatment for breast cancer. Herbal plant Dillenia suffruticosa (Griff) Martelli under the family Dilleniaceae has been traditionally used to treat cancerous growth. In this study, the anticancer effect of ethyl acetate extract of D. suffruticosa (EADs) was examined on human breast adenocarcinoma cell line MCF-7 and the molecular pathway involved was elucidated.
Methods
EADs was obtained from the root of D. suffruticosa by using sequential solvent extraction. Cytotoxicity was determined by using MTT assay, mode of cell death by cell cycle analysis and apoptosis induction by Annexin-FITC/PI assay. Morphology changes in cells were observed under inverted light microscope. Involvement of selected genes in the oxidative stress-mediated signaling pathway was explored using multiplex gene expression analysis.
Results
The treatment of EADs caused cytotoxicity to MCF-7 cells in a dose- and time-dependent manner at 24, 48 and 72 hours with IC50 of 76 ± 2.3, 58 ± 0.7 and 39 ± 3.6 μg/mL, respectively. The IC50 of tamoxifen-treated MCF-7 cells was 8 ± 0.5 μg/mL. Induction of apoptosis by EADs was dose- and time- dependent. EADs induced non-phase specific cell cycle arrest at different concentration and time point. The multiplex mRNA expression study indicated that EADs-induced apoptosis was accompanied by upregulation of the expression of SOD1, SOD2, NF-κB, p53, p38 MAPK, and catalase, but downregulation of Akt1.
Conclusion
It is suggested that EADs induced apoptosis in MCF-7 cells by modulating numerous genes which are involved in oxidative stress pathway. Therefore, EADs has the potential to act as an effective intervention against breast cancer cells.
doi:10.1186/1472-6882-14-55
PMCID: PMC3927215  PMID: 24524627
Dillenia suffruticosa; Breast cancer; Cytotoxic; Apoptosis; Oxidative stress pathway
23.  Effect of pheniramine maleate on reperfusion injury in brain tissue 
Background
The aim of this study was to investigate the protective effects of methylprednisolone (Pn), which is a potent anti-inflammatory agent, and pheniramine maleate (Ph), which is an antihistaminic with some anti-inflammatory effects, on reperfusion injury in brain developing after ischemia of the left lower extremity of rats.
Material/Methods
Twenty-eight randomly selected male Sprague-Dawley rats were divided into 4 groups: Group 1 was the control group, Group 2 was the sham group (I/R), Rats in Group 3 were subjected to I/R and given Ph, and rats in Group 4 were subjected to I/R and given Pn. A tourniquet was applied at the level of left groin region of subjects in the I/R group after induction of anesthesia. One h of ischemia was performed with no drug administration. In the Ph group, half of a total dose of 10 mg/kg Ph was administered intraperitoneally before ischemia and the remaining half before reperfusion. In the Pn group, subjects received a single dose of 50 mg/kg Pn intraperitoneally at the 30th min of ischemia. Brains of all subjects were removed after 24 h for examination.
Results
Malondialdehyde (MDA) levels of the prefrontal cortex were significantly lower in the Ph group than in the I/R group (p<0.05). Superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzyme activities were found to be significantly higher in the Ph group than in the I/R group (p<0.05). Histological examination demonstrated that Ph had protective effects against I/R injury developing in the brain tissue.
Conclusions
Ph has a protective effect against ischemia/reperfusion injury created experimentally in rat brains.
doi:10.12659/MSMBR.889570
PMCID: PMC3867399  PMID: 24309384
ischemia/reperfusion injury; pheniramine maleate; brain
24.  Noise-Induced Hearing Loss Among Quarry Workers in a North-Eastern State of Malaysia: A Study on Knowledge, Attitude and Practice 
Oman Medical Journal  2013;28(5):331-336.
Objectives
Noise is known to be one of the environmental and occupational hazards listed in the Factory and Machinery Act 1967. Quarries with loud deafening sounds from trucks and machineries pose the risk of noise-induced hearing loss to workers. This study was designed to assess the knowledge, attitude and practice towards noise-induced hearing loss and to determine the prevalence of noise-induced hearing loss and its associated factors among quarry workers in a north-eastern state of Malaysia.
Methods
A cross-sectional study was conducted at six quarries in a north-eastern state of Malaysia, with 97 consented respondents who answered a validated version of a questionnaire and underwent pure tone audiogram. The respondents were male, aged between 18 to 50 years, working in the quarry area for at least 6-months duration with no family history of ear diseases.
Results
The mean percentage scores of knowledge, attitude and practice were 44 (11), 70 (10) and 28 (16) percent, respectively. The prevalence of noise-induced hearing loss was found to be 57 (95% CI: 47, 67) with 46 (84%) having mild and moderate noise-induced hearing loss, and 34 (62%) involved both ears. Multiple logistic regressions showed that age and practice score were the associated factors with odd ratios of 1.1 (95% CI: 1.1, 1.2; p<0.001) and 0.9 (95% CI: 0.8, 1.0; p=0.008), respectively.
Conclusion
The knowledge, attitude and practice scores of the respondents were poor and the high prevalence of noise-induced hearing loss was contributed by factors such as poor practice and old age.
doi:10.5001/omj.2013.96
PMCID: PMC3769125  PMID: 24044059
Noise-induced hearing loss; Knowledge, attitude and practice; Prevalence; Quarry workers
25.  Ethyl acetate extract of germinated brown rice attenuates hydrogen peroxide-induced oxidative stress in human SH-SY5Y neuroblastoma cells: role of anti-apoptotic, pro-survival and antioxidant genes 
Background
There are reports of improved metabolic outcomes due to consumption of germinated brown rice (GBR). Many of the functional effects of GBR can be linked to its high amounts of antioxidants. Interestingly, dietary components with high antioxidants have shown promise in the prevention of neurodegenerative diseases like Alzheimer’s disease (AD). This effect of dietary components is mostly based on their ability to prevent apoptosis, which is believed to link oxidative damage to pathological changes in AD. In view of the rich antioxidant content of GBR, we studied its potential to modulate processes leading up to AD.
Methods
The total phenolic content and antioxidant capacity of the ethyl acetate extract of GBR were compared to that of brown rice (BR), and the cytotoxicity of both extracts were determined on human SH-SY5Y neuronal cells using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) Assay. Based on its higher antioxidant potentials, the effect of the GBR extract on morphological changes due to hydrogen peroxide (H2O2)-induced oxidative damage in human SH-SY5Y neuronal cells was examined using inverted light microscope and fluorescence microscope by means of acridine orange-propidium iodide (AO/PI) staining. Also, evaluation of the transcriptional regulation of antioxidant and apoptotic genes was carried out using Multiplex Gene Expression System.
Results
The ethyl acetate extract of GBR had higher total phenolic content and antioxidant capacity compared to BR. The cytotoxicity results showed that GBR extract did not cause any damage to the human SH-SY5Y neuronal cells at concentrations of up to 20 ppm, and the morphological analyses showed that the GBR extract (up to 10 ppm) prevented H2O2-induced apoptotic changes in the cells. Furthermore, multiplex gene expression analyses showed that the protection of the cells by the GBR extract was linked to its ability to induce transcriptional changes in antioxidant (SOD 1, SOD 2 and catalase) and apoptotic (AKT, NF-Kβ, ERK1/2, JNK, p53 and p38 MAPK) genes that tended towards survival.
Conclusions
Taken together, the results of our study showed that the ethyl acetate extract of GBR, with high antioxidant potentials, could prevent H2O2-induced oxidative damage in SH-SY5Y cells. The potential of GBR and its neuroprotective mechanism in ameliorating oxidative stress-related cytotoxicity is therefore worth exploring further.
doi:10.1186/1472-6882-13-177
PMCID: PMC3726323  PMID: 23866310
Germinated brown rice; Antioxidant; Oxidative stress; Neuroprotective; SH-SY5Y

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