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Fulltext Independent Emergence of the Cosmopolitan Asian Chikungunya Virus, Philippines 2012

Tan KK, Sy AK, Tandoc AO, Khoo JJ, Sulaiman S, Chang LY, et al.

Sci Rep, 2015 Jul 23;5:12279.
PMID: 26201250 DOI: 10.1038/srep12279

Outbreaks involving the Asian genotype Chikungunya virus (CHIKV) caused over one million infections in the Americas recently. The outbreak was preceded by a major nationwide outbreak in the Philippines. We examined the phylogenetic and phylogeographic relationships of representative CHIKV isolates obtained from the 2012 Philippines outbreak with other CHIKV isolates collected globally. Asian CHIKV isolated from the Philippines, China, Micronesia and Caribbean regions were found closely related, herein denoted as Cosmopolitan Asian CHIKV (CACV). Three adaptive amino acid substitutions in nsP3 (D483N), E1 (P397L) and E3 (Q19R) were identified among CACV. Acquisition of the nsP3-483N mutation in Compostela Valley followed by E1-397L/E3-19R in Laguna preceded the nationwide spread in the Philippines. The China isolates possessed two of the amino acid substitutions, nsP3-D483N and E1-P397L whereas the Micronesian and Caribbean CHIKV inherited all the three amino acid substitutions. The unique amino acid substitutions observed among the isolates suggest multiple independent virus dissemination events. The possible biological importance of the specific genetic signatures associated with the rapid global of the virus is not known and warrant future in-depth study and epidemiological follow-up. Molecular evidence, however, supports the Philippines outbreak as the possible origin of the CACV.
Fulltext Incidental pharmacogenetics findings in an HLA-related research: Considerations for primary prevention

Bakhtiar MF, Too CL, Tang MM, Sulaiman S, Tan LK, Ahmad-Fauzi NA, et al.

Clin Exp Allergy, 2019 04;49(4):537-540.
PMID: 30693574 DOI: 10.1111/cea.13347
Fulltext In vitro whole blood clot lysis for fibrinolytic activity study using d-dimer and confocal microscopy

Elnager A, Abdullah WZ, Hassan R, Idris Z, Wan Arfah N, Sulaiman SA, et al.

Adv Hematol, 2014;2014:814684.
PMID: 24660000 DOI: 10.1155/2014/814684

This study aimed to evaluate in vitro whole blood (WB) clot lysis method for the assessment of fibrinolytic activity. Standardized unresected (uncut) retracted WB clot was incubated in pool platelet poor plasma (PPP) for varying incubation times and in streptokinase (SK) at different concentrations. The fibrinolytic activity was assessed by D-dimer (DD), confocal microscopy, and clot weight. DD was measured photometrically by immunoturbidimetric method. There was a significant difference in mean DD levels according to SK concentrations (P = 0.007). The mean DD ± SD according to the SK concentrations of 5, 30, 50, and 100 IU/mL was: 0.69 ± 0.12, 0.78 ± 0.14, 1.04 ± 0.14 and 2.40 ± 1.09 μ g/mL. There were no significant changes of clot weight at different SK concentrations. Gradual loss and increased branching of fibrin in both PPP and SK were observed. Quantitation of DD and morphology of fibrin loss as observed by the imaging features are in keeping with fibrinolytic activity. Combination of DD levels and confocal microscopic features was successfully applied to evaluate the in vitro WB clot lysis method described here.
In vitro ovicidal and larvicidal activity of methanolic leaf extract of Manihot esculenta (cassava) on susceptible and resistant strains of Trichostrongylus colubriformis

Al-Rofaai A, Rahman WA, Sulaiman SF, Yahaya ZS

Vet Parasitol, 2012 Nov 23;190(1-2):127-35.
PMID: 22749290 DOI: 10.1016/j.vetpar.2012.05.028

This study aimed to represent the first report of the ovicidal and larvicidal activity of the methanolic leaf extract of Manihot esculenta (cassava) against eggs and larvae of susceptible and resistant strains of Trichostrongylus colubriformis. As well as, to determine the total tannin compounds, antioxidant activity and toxicity of the extract. The egg hatch test was used to evaluate ovicidal activity against unembryonated eggs, whereas larval feeding inhibition assay and MTT-formazan assay were used to evaluate larvicidal activity against first (L(1)) and infective (L(3)) larvae, respectively. The results showed no significant differences were detected between the sensitivities of susceptible and resistant strains of T. colubriformis to the extract. Eggs, L(1) and L(3) were significantly affected (P<0.001) compared with negative control, and L(1) were more sensitive than the eggs and L(3). The total tannin compounds were investigated using tannin quantification assay and determined by 254.44 TAE/mg. The antioxidant activity was evaluated using the DPPH radical scavenging assay and the median inhibition concentration (IC(50)) was determined by 2.638 mg/ml. Acute oral toxicity at dose of 5,000 mg/kg, and sub-chronic oral toxicity at 500 and 1,000 mg/kg of the extract were observed in male and female Sprague-Dawley (SD) rats. The acute oral toxicity revealed that the median lethal dose (LD(50)) of methanolic extract of cassava leaves on SD rats was greater than 5,000 mg/kg, whereas the sub-chronic oral toxicity did not show observed adverse effects at 500 and 1,000 mg/kg per day for 28 days. In conclusion, the methanolic extract of cassava leaves has direct ovicidal and larvicidal activity against T. colubriformis strains with a safety margin for animals, and it may be potentially utilized as a source of natural antioxidants.
In vitro activity of neem (Azadirachta indica) and cassava (Manihot esculenta) on three pre-parasitic stages of susceptible and resistant strains of Teladorsagia (Ostertagia) circumcincta

Al-Rofaai A, Rahman WA, Sulaiman SF, Yahaya ZS

Vet Parasitol, 2012 Aug 13;188(1-2):85-92.
PMID: 22455724 DOI: 10.1016/j.vetpar.2012.03.002

Anthelmintic resistance of gastrointestinal nematodes is considered as one of the main limiting factors causing significant economic losses to the small ruminant industry. The anthelmintic properties of some plants are among the suggested alternative solutions to control these parasitic worms. The present study investigated the anthelmintic activity of neem (Azadirachta indica) and cassava (Manihot esculenta) leaf extracts against the susceptible and resistant strains of one of the most important nematodes in small ruminants, Teladorsagia (Ostertagia) circumcincta. Three different in vitro tests: egg hatch test, larval development assay, and larval paralysis assay were used to determine the efficiency of neem and cassava extracts on three pre-parasitic stages of T. circumcincta. The LC(50) was determined for the most potent extract in each plant as well as the phytochemical tests, total tannin quantification and cytotoxicity on peripheral blood mononuclear cells of goats. The results revealed a high anthelmintic activity of neem methanol extract (NME) and cassava methanol extract (CME) on both strains of T. circumcincta without significant differences between the strains. The first stage larvae were more sensitive with the lowest LC(50) at 7.15 mg/ml and 10.72 mg/ml for NME and CME, respectively, compared with 44.20mg/ml and 56.68 mg/ml on eggs and 24.91 mg/ml and 71.96 mg/ml on infective stage larvae.
In situ transesterification of solid coconut waste in a packed bed reactor with CaO/PVA catalyst

Talha NS, Sulaiman S

Waste Manag, 2018 Aug;78:929-937.
PMID: 32559988 DOI: 10.1016/j.wasman.2018.07.015

In this study, solid coconut waste and CaO/PVA was used as raw material and catalyst respectively to produce biodiesel through in situ transesterification. Both, raw material and catalyst were packed in a packed bed reactor. The reaction was fixed for 3 h and the mixing was kept constant at 350 rpm. The highest biodiesel yield of 95% was obtained at reaction temperature of 61 °C with catalyst loading (CaO/PVA) of 2.29 wt% and methanol to solid ratio of 12:1. CaO-waste derived catalyst has been successfully proven to be utilized as heterogeneous base catalyst for the production of biodiesel from solid coconut waste.
Impact of healthy eating practices and physical activity on quality of life among breast cancer survivors

Mohammadi S, Sulaiman S, Koon PB, Amani R, Hosseini SM

Asian Pac J Cancer Prev, 2013;14(1):481-7.
PMID: 23534778

Following breast cancer diagnosis, women often attempt to modify their lifestyles to improve their health and prevent recurrence. These behavioral changes typically involve diet and physical activity modification. The aim of this study was to determine association between healthy eating habits and physical activity with quality of life among Iranian breast cancer survivors. A total of 100 Iranian women, aged between 32 to 61 years were recruited to participate in this cross-sectional study. Eating practices were evaluated by a validated questionnaire modified from the Women's Healthy Eating and Living (WHEL) study. Physical activity was assessed using the International Physical Activity Questionnaire (IPAQ). A standardized questionnaire by the European Organization of Research and Treatment of Cancer Quality of Life and its breast cancer module (EORTC QLQ-C30/+BR-23) were applied to determine quality of life. Approximately 29% of the cancer survivors were categorized as having healthy eating practices, 34% had moderate eating practices and 37% had poor eating practices based on nutrition guidelines. The study found positive changes in the decreased intake of fast foods (90%), red meat (70%) and increased intake of fruits (85%) and vegetables (78%). Generally, breast cancer survivors with healthy eating practices had better global quality of life, social, emotional, cognitive and role functions. Result showed that only 12 women (12%) met the criteria for regular vigorous exercise, 22% had regular moderate-intensity exercise while the majority (65%) had low-intensity physical activity. Breast cancer survivors with higher level of physical activity had better emotional and cognitive functions. Healthy eating practices and physical activity can improve quality of life of cancer survivors. Health care professionals should promote good dietary habits and physical activity to improve survivors' health and quality of life.
Fulltext Impact of Hepatitis B on Human Immunodeficiency Virus Patients in Malaysia: A Retrospective Study

Khan AH, Syed Sulaiman SA, Akhtar A, Adnan AS, Aftab RA

Value Health, 2014 Nov;17(7):A803.
PMID: 27203024 DOI: 10.1016/j.jval.2014.08.505
Fulltext Impact of Cigarette and Alcohol Use on Adverse Drug Reactions of Haart Therapy Among HIV/AIDS Patients

Khan AH, Syed Sulaiman SA, Khan K, Soo CT, Akhtar A

Value Health, 2014 Nov;17(7):A802-3.
PMID: 27203019 DOI: 10.1016/j.jval.2014.08.503
Fulltext Immunogenicity and Safety of a Tetravalent Dengue Vaccine Administered Concomitantly or Sequentially With Quadrivalent Human Papillomavirus Vaccine in Boys and Girls 9-13 Years of Age in Malaysia: A Phase IIIb, Randomized, Open-label Study

Hassan J, Toh TH, Sivapunniam SK, Hasim R, Ghazali NF, Sulaiman S, et al.

Pediatr Infect Dis J, 2021 08 01;40(8):774-781.
PMID: 34250977 DOI: 10.1097/INF.0000000000003164

BACKGROUND: Incorporating dengue vaccination within existing vaccination programs could help improve dengue vaccine coverage. We assessed the immunogenicity and safety of a quadrivalent human papillomavirus (HPV) vaccine administered concomitantly or sequentially with a tetravalent dengue vaccine (CYD-TDV) in healthy children 9-13 years of age in Malaysia.
METHODS: In this phase IIIb, open-label, multicenter study (NCT02993757), participants were randomized 1:1 to receive 3 CYD-TDV doses 6 months apart and 2 doses of quadrivalent HPV vaccine concomitantly with, or 1 month before (sequentially), the first 2 CYD-TDV doses. Only baseline dengue-seropositive participants received the 3 doses. Antibody levels were measured at baseline and 28 days after each injection using an enzyme-linked immunosorbent assay for HPV-6, -9, -16 and -18, and the 50% plaque reduction neutralization test for the 4 dengue serotypes; immunogenicity results are presented for baseline dengue-seropositive participants. Safety was assessed throughout the study for all participants.
RESULTS: At baseline, 197 of 528 (37.3%) randomized participants were dengue-seropositive [n = 109 (concomitant group) and n = 88 (sequential group)]. After the last HPV vaccine dose, antibody titers for HPV among baseline dengue-seropositive participants were similar between treatment groups, with between-group titer ratios close to 1 for HPV-6 and 0.8 for HPV-11, -16, and -18. After CYD-TDV dose 3, dengue antibody titers were similar between treatment groups for all serotypes [between-group ratios ranged from 0.783 (serotype 2) to 1.07 (serotype 4)]. No safety concerns were identified.
CONCLUSIONS: The immunogenicity and safety profiles of CYD-TDV and quadrivalent HPV vaccines were unaffected when administered concomitantly or sequentially in dengue-seropositive children.
Immune-stimulating properties of 80% methanolic extract of Ludwigia octovalvis against Shiga toxin-producing E. coli O157:H7 in Balb/c mice following experimental infection

Kadum Yakob H, Manaf Uyub A, Fariza Sulaiman S

J Ethnopharmacol, 2015 Aug 22;172:30-7.
PMID: 26091966 DOI: 10.1016/j.jep.2015.06.006

Ludwigia octovalvis is an aquatic plant widely distributed throughout the tropical and sub-tropical regions. It is commonly consumed as a health drink and traditionally used for treating various ailments such as dysentery, diarrhea, diabetes, nephritisn and headache. No information is available on its in vivo antibacterial activity against an important foodborne pathogen, Shiga toxin producing Escherichia coli O157:H7.
Immediate compared with delayed oxytocin after amniotomy labor induction in parous women: a randomized controlled trial

Tan PC, Soe MZ, Sulaiman S, Omar SZ

Obstet Gynecol, 2013 Feb;121(2 Pt 1):253-259.
PMID: 23344273 DOI: 10.1097/AOG.0b013e31827e7fd9

OBJECTIVE: To compare immediate with delayed (4 hours) oxytocin infusion after amniotomy on vaginal delivery within 12 hours and patient satisfaction with the birth process.
METHODS: Parous women with favorable cervixes after amniotomy for labor induction were randomized to immediate titrated oxytocin or placebo intravenous infusion in a double-blind noninferiority trial. After 4 hours, study infusions were stopped, the women were assessed, and open-label oxytocin was started if required. Maternal satisfaction with the birth process was assessed with a 10-point visual numerical rating scale (lower score, greater satisfaction).
RESULTS: Vaginal delivery rates at 12 hours were 91 of 96 (94.8%) compared with 91 of 94 (96.8%) (relative risk 0.98, 95% confidence interval [CI] 0.92-1.04, P=.72), and maternal satisfaction on a visual numerical rating scale (median [interquartile range]) was 3 [3-4] compared with 3 [3-5], P=.36 for immediate compared with delayed arm, respectively). Cesarean delivery, maternal fever, postpartum hemorrhage, uterine hyperactivity, and adverse neonatal outcome rates were similar between arms. The immediate oxytocin arm had a shorter amniotomy-to-delivery interval of 5.3±3.1 compared with 6.9±2.9 hours (P
Fulltext Identification of sRNA mediated responses to nutrient depletion in Burkholderia pseudomallei

Mohd-Padil H, Damiri N, Sulaiman S, Chai SF, Nathan S, Firdaus-Raih M

Sci Rep, 2017 12 07;7(1):17173.
PMID: 29215024 DOI: 10.1038/s41598-017-17356-4

The Burkholderia genus includes many species that are known to survive in diverse environmental conditions including low nutrient environments. One species, Burkholderia pseudomallei is a versatile pathogen that can survive in a wide range of hosts and environmental conditions. In this study, we investigated how a nutrient depleted growth environment evokes sRNA mediated responses by B. pseudomallei. Computationally predicted B. pseudomallei D286 sRNAs were mapped to RNA-sequencing data for cultures grown under two conditions: (1) BHIB as a nutrient rich media reference environment and (2) M9 media as a nutrient depleted stress environment. The sRNAs were further selected to identify potentially cis-encoded systems by investigating their possible interactions with their flanking genes. The mappings of predicted sRNA genes and interactions analysis to their flanking genes identified 12 sRNA candidates that may possibly have cis-acting regulatory roles that are associated to a nutrient depleted growth environment. Our approach can be used for identifying novel sRNA genes and their possible role as cis-mediated regulatory systems.
Fulltext Hypertension Treatment in Chronic Kidney Disease Stage 3, 4 and 5: A Hospital Based Cross-Sectional Study in Malaysia

Adnan AS, Salman M, Sulaiman SA, Khan AH, Hamzah DA

Value Health, 2014 Nov;17(7):A757.
PMID: 27202763 DOI: 10.1016/j.jval.2014.08.237
Human amniotic membrane as a chondrocyte carrier vehicle/substrate: in vitro study

Krishnamurithy G, Shilpa PN, Ahmad RE, Sulaiman S, Ng CL, Kamarul T

J Biomed Mater Res A, 2011 Dec 01;99(3):500-6.
PMID: 21913317 DOI: 10.1002/jbm.a.33184

Human amniotic membrane (HAM) is an established biomaterial used in many clinical applications. However, its use for tissue engineering purposes has not been fully realized. A study was therefore conducted to evaluate the feasibility of using HAM as a chondrocyte substrate/carrier. HAMs were obtained from fresh human placenta and were process to produced air dried HAM (AdHAM) and freeze dried HAM (FdHAM). Rabbit chondrocytes were isolated and expanded in vitro and seeded onto these preparations. Cell proliferation, GAG expression and GAG/cell expression were measured at days 3, 6, 9, 12, 15, 21, and 28. These were compared to chondrocytes seeded onto plastic surfaces. Histological analysis and scanning electron microscopy was performed to observe cell attachment. There was significantly higher cell proliferation rates observed between AdHAM (13-51%, P=0.001) or FdHAM (18-48%, p = 0.001) to chondrocytes in monolayer. Similarly, GAG and GAG/cell expressed in AdHAM (33-82%, p = 0.001; 22-60%, p = 0.001) or FdHAM (41-81%, p = 0.001: 28-60%, p = 0.001) were significantly higher than monolayer cultures. However, no significant differences were observed in the proliferation rates (p = 0.576), GAG expression (p = 0.476) and GAG/cell expression (p = 0.135) between AdHAM and FdHAM. The histology and scanning electron microscopy assessments demonstrates good chondrocyte attachments on both HAMs. In conclusion, both AdHAM and FdHAM provide superior chondrocyte proliferation, GAG expression, and attachment than monolayer cultures making it a potential substrate/carrier for cell based cartilage therapy and transplantation.
Human amnion as a novel cell delivery vehicle for chondrogenic mesenchymal stem cells

Tan SL, Sulaiman S, Pingguan-Murphy B, Selvaratnam L, Tai CC, Kamarul T

Cell Tissue Bank, 2011 Feb;12(1):59-70.
PMID: 19953328 DOI: 10.1007/s10561-009-9164-x

This study investigates the feasibility of processed human amnion (HAM) as a substrate for chondrogenic differentiation of mesenchymal stem cells (MSCs). HAM preparations processed by air drying (AD) and freeze drying (FD) underwent histological examination and MSC seeding in chondrogenic medium for 15 days. Monolayer cultures were used as control for chondrogenic differentiation and HAMs without cell seeding were used as negative control. Qualitative observations were made using scanning electron microscopy analysis and quantitative analyses were based on the sulfated glycosaminoglycans (GAG) assays performed on day 1 and day 15. Histological examination of HAM substrates before seeding revealed a smooth surface in AD substrates, while the FD substrates exhibited a porous surface. Cell attachment to AD and FD substrates on day 15 was qualitatively comparable. GAG were significantly highly expressed in cells seeded on FD HAM substrates. This study indicates that processed HAM is a potentially valuable material as a cell-carrier for MSC differentiation.
High 5-hydroxymethylfurfural concentrations are found in Malaysian honey samples stored for more than one year

Khalil MI, Sulaiman SA, Gan SH

Food Chem Toxicol, 2010 Aug-Sep;48(8-9):2388-92.
PMID: 20595027 DOI: 10.1016/j.fct.2010.05.076

5-Hydroxymethylfurfural (HMF) content is an indicator of the purity of honey. High concentrations of HMF in honey indicate overheating, poor storage conditions and old honey. This study investigated the HMF content of nine Malaysian honey samples, as well as the correlation of HMF formation with physicochemical properties of honey. Based on the recommendation by the International Honey Commission, three methods for the determination of HMF were used: (1) high performance liquid chromatography (HPLC), (2) White spectrophotometry and (3) Winkler spectrophotometry methods. HPLC and White spectrophotometric results yielded almost similar values, whereas the Winkler method showed higher readings. The physicochemical properties of honey (pH, free acids, lactones and total acids) showed significant correlation with HMF content and may provide parameters that could be used to make quick assessments of honey quality. The HMF content of fresh Malaysian honey samples stored for 3-6 months (at 2.80-24.87 mg/kg) was within the internationally recommended value (80 mg/kg for tropical honeys), while honey samples stored for longer periods (12-24 months) contained much higher HMF concentrations (128.19-1131.76 mg/kg). Therefore, it is recommended that honey should generally be consumed within one year, regardless of the type.
Healthy eating index and breast cancer risk among Malaysian women

Shahril MR, Sulaiman S, Shaharudin SH, Akmal SN

Eur J Cancer Prev, 2013 Jul;22(4):342-7.
PMID: 23702680 DOI: 10.1097/CEJ.0b013e32835b37f9

Healthy Eating Index-2005 (HEI-2005), an index-based dietary pattern, has been shown to predict the risk of chronic diseases among Americans. This study aims to examine the ability of HEI-2005 in predicting the probability for risk of premenopausal and postmenopausal breast cancer among Malaysian women. Data from a case-control nutritional epidemiology study among 764 participants including 382 breast cancer cases and 382 healthy women were extracted and scored. Multivariate odds ratios (OR) with 95% confidence intervals (CI) were used to evaluate the relationship between the risk of breast cancer and quartiles (Q) of HEI-2005 total scores and its component, whereas the risk prediction ability of HEI-2005 was investigated using diagnostics analysis. The results of this study showed that there is a significant reduction in the risk of breast cancer, with a higher HEI-2005 total score among premenopausal women (OR Q1 vs. Q4=0.34, 95% CI; 0.15-0.76) and postmenopausal women (OR Q1 vs. Q4=0.20, 95% CI; 0.06-0.63). However, HEI-2005 has a sensitivity of 56-60%, a specificity of 55-60%, and a positive predictive value and negative predictive value of 57-58%, which indicates a moderate ability to predict the risk of breast cancer according to menopausal status. The breast cancer incidence observed poorly agrees with risk outcomes from HEI-2005 as shown by low κ statistics (κ=0.15). In conclusion, although the total HEI-2005 scores were associated with a risk of breast cancer among Malaysian women, the ability of HEI-2005 to predict risk is poor as indicated by the diagnostic analysis. A local index-based dietary pattern, which is disease specific, is required to predict the risk of breast cancer among Malaysian women for early prevention.
Fulltext Guideline adherence and control of diabetes mellitus with co-morbidities in a tertiary-care hospital in Malaysia [Abstract]

Iqbal MZ, Iqbal MS, Khan AH, Sulaiman SA, Iqbal MW

Value Health, 2014 Nov;17(7):A353.
PMID: 27200693 DOI: 10.1016/j.jval.2014.08.744

Conference abstract:
OBJECTIVES.. To evaluate doctors’ adherence to Malaysian Clinical Practice Guideline (CPG) 2009 in the management of diabetes mellitus with co-morbidities in Malaysia.
METHODS. Cross-sectional study was done at a tertiary-care hospital in Malaysia. Total 51 physicians and 1020 patients’ prescriptions written by same physicians (20 prescriptions per physician) were analyzed. All patients had diabetes mellitus with co-morbidities. Depending on the recommendations of CPG 2009, the prescriptions were clustered as adherent and non-adherent prescriptions. All obtained data were analyzed using descriptive and inferential statistics.
RESULTS. A statistically significant negative association (Ф= 0.094, p-value=0.003) was observed between diabetes mellitus control and co-morbidities. CPG adherent had statistically weak negative association (Ф= - 0.081, p-value=0.010) with patients having co-morbidities (41.6%). No statistically significant association was observed between CPG adherence and any other co-morbidity. Majority of the patients received guidelines-compliant pharmacotherapy. The overall good level of physician adherence with CPG 2009 was observed in the management of diabetes mellitus with co-morbidities.
CONCLUSIONS. The study explored several features of prescription pattern of the physicians involved in the management of diabetes mellitus with co-morbidities and recognized the need for improvement in their prescription pattern for treating the diabetes mellitus.
Growth arrest and non-apoptotic programmed cell death associated with the up-regulation of c-myc mRNA expression in T-47D breast tumor cells following exposure to Epipremnum pinnatum (L.) Engl. hexane extract

Tan ML, Muhammad TS, Najimudin N, Sulaiman SF

J Ethnopharmacol, 2005 Jan 15;96(3):375-83.
PMID: 15619555

Epipremnum pinnatum (L.) Engl. hexane extract produced a significant growth inhibition against T-47D breast carcinoma cells and analysis of cell death mechanisms indicated that the extract elicited a non-apoptotic programmed cell death. T-47D cells exposed to the extract at EC(50) concentration (72 h) for 24 h failed to demonstrate typical DNA fragmentation associated with apoptosis, as carried out using a modified TUNEL assay. In addition, acute exposure to the extract produced an insignificant regulation of caspase-3 and p53 mRNA expression but increased in the c-myc mRNA expression. Ultrastructural analysis using transmission electron microscope demonstrated distinct vacuolated cells, which strongly indicated a Type II non-apoptotic cell death although the changes in chromatin were also detected. The presence of non-apoptotic programmed cell death was then reconfirmed with annexin-V and propidium iodide staining. These findings suggested that up-regulation of c-myc mRNA expression may have contributed to the growth arrest and Type II non-apoptotic programmed cell death in the Epipremnum pinnatum (L.) Engl. hexane extract-treated T-47D cells.

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