Comparison of cytotoxicity between extracts of Clinacanthus nutans (Burm. f.) Lindau leaves from different locations and the induction of apoptosis by the crude methanol leaf extract in D24 human melanoma cells.

BACKGROUND: Clinacanthus nutans (Burm. f.) Lindau leaves are widely used by cancer patients and the leaf extracts possess cytotoxic and antiproliferative effects on several human cancer cell lines. However, the effect of C. nutans leaf extract on human melanoma, which is the least common but most fatal form of skin cancer and one of the most common cancers diagnosed in both sexes worldwide, is unknown. There is also limited information on whether the bioactivity of extracts differs between C. nutans leaves grown in different geographical locations with varying environmental conditions. METHODS: The present study, for the first time, compared and demonstrated the cytotoxicity of the crude methanol extracts of C. nutans leaves from 11 different locations in Malaysia, Thailand and Vietnam, with diverse environmental conditions against D24 melanoma cells through WST-8 assay. The percentage of apoptotic cells following treatment with the most active extract was determined in a dose- and time-dependent manner by a cytofluorometric double staining technique. Biochemical and morphological changes in the treated and untreated cells were examined by fluorescence and transmission electron microscopy techniques, respectively, to further affirm the induction of apoptosis. RESULTS: The leaves of plants grown at higher elevations and lower air temperatures were more cytotoxic to the D24 melanoma cells than those grown at lower elevations and higher air temperatures, with the leaf extract from Chiang Dao, Chiang Mai, Thailand exhibited the highest cytotoxicity (24 h EC50: 0.95 mg/mL and 72 h EC50: 0.77 mg/mL). This most active crude extract induced apoptotic cell death in the D24 cells in a dose- and time-dependent manner. Typical biochemical and morphological characteristics of apoptosis were also observed in the treated D24 cells. CONCLUSIONS: The results, showing the cytotoxicity of C. nutans and the induction of apoptosis in D24 cells, are significant and useful to facilitate the development of C. nutans as a potential novel chemotherapeutic agent for the management of skin melanoma.

Pharmacological properties of Hoya (Apocynaceae): a systematic review.

In tropical forests, Hoya, a plant with significant indigenous medicinal applications, has been underexplored in pharmacological studies. This systematic review meticulously investigates the diverse pharmacological effects exhibited by various Hoya species on human health. A comprehensive literature search, encompassing Scopus, ScienceDirect, and SpringerLink databases, employed specific keyword combinations ('Hoya' and 'pharmacological properties' OR 'pharmacology property'). The included studies exclusively focused on Hoya's impact on human health. The findings underscore Hoya's potential as a medicinal plant, demonstrating promising attributes such as anticancer, antibacterial, antioxidant, anti-inflammatory, anti-diabetic, antinociceptive, and parasympatholytic effects. Despite these promising indications, the review underscores the necessity for further in vivo investigations to fully unlock Hoya's therapeutic potential. A comprehensive understanding of its mechanisms of action, efficacy, and safety in living systems is imperative for realising its holistic therapeutic benefits.

The incidence, and spatial trends of cholera in Sabah over 15 years: Repeated outbreaks in coastal areas.

Vibrio cholerae remains a notable public health challenge across Malaysia. Although the Malaysian state of Sabah is considered a cholera-affected area, gaps remain in understanding the epidemiological trends and spatial distribution of outbreaks. Therefore, to determine longitudinal and spatial trends in cholera cases data were obtained from the Sabah State Health Department for all notified cases of cholera between 2005-2020. A cholera outbreak is defined as one or more confirmed cases in a single locality with the evidence of local transmission. All records were geolocated to village level. Satellite-derived data and generalised linearized models were used to assess potential risk factors, including population density, elevation, and distance to the sea. Spatiotemporal clustering of reported cholera cases and zones of increased cholera risk were evaluated using the tau statistic (τ) at 550m, 5km and 10km distances. Over a 15-year period between 2005-2020, 2865 cholera cases were recorded in Sabah, with a mean incidence rate of 5.6 cases per 100,000 (95% CI: 3.4-7.9). From 2015-2020, 705 symptomatic cases and 727 asymptomatic cases were reported. Symptomatic cases primarily occurred in local Malaysian populations (62.6%, 441/705) and in children and adolescents under 15-years old (49.4%, 348/705). On average, cases were reported in areas with low population density (19.45 persons/km2), low elevations (19.45m) and near coastal areas. Spatiotemporal clustering of cholera cases was identified up to 3.5km, with increased village-level cholera risk within 500m and 5 days of initial case presentation to a health facility (Risk Ratio = 9.7, 95% CI: 7.5-12.4). Cholera incidence has high spatial and temporal heterogeneity within Sabah, with some districts experiencing repeated outbreaks. Cholera cases clustered across space and time, with village-level risk of cholera highest within 5 days and within close proximity to primary case villages, suggesting local transmission.

Association between Leptin (G2548A) and Leptin Receptor (Q223R) Polymorphisms with Plasma Leptin, BMI, Stress, Sleep and Eating Patterns among the Multiethnic Young Malaysian Adult Population from a Healthcare University.

Relative leptin resistance in childhood to absolute leptin resistance in maturity suggests sleep, eating behaviour, and the psychological state as probable causes. The current body of research provides inconclusive evidence linking G2548A and Q223R to obesity. Furthermore, we could find very little data that have observed the association between the environment and gene polymorphism, especially in the multiethnic population that exists in Malaysia. This study searched for a possible link between sleeping habits, eating behaviour, and stress indicators with plasma leptin and its genetic variation in young adult Malaysian healthcare students. The study involved 185 first- and second-year medical and dental students from a healthcare university. Polymerase Chain Reaction−Restriction Fragment Length Polymorphism(PCR-RFLP) determined the genotype, Enzyme Linked Immunoabsorbant Assay (ELISA) tested the serum leptin, and a self-administered questionnaire evaluated sleep, eating behaviour, and psychological condition. Gender and ethnicity are linked to fasting plasma leptin levels (p < 0.001). Plasma leptin also affects stress, anxiety, and sadness. Leptin (LEP) and Leptin Receptor (LEPR) polymorphisms were not associated with BMI, plasma leptin, sleep, eating behaviour, or psychological state. Young adult Malaysian Indians were obese and overweight, while Chinese were underweight. These findings imply overweight and obese participants were in stage I of leptin resistance and lifestyle change or leptin therapy could prevent them from becoming cripplingly obese as they age.

Mumps outbreak in university students: first detection of mumps virus genotype F in Borneo.

BACKGROUND: In October 2016, a mumps outbreak occurred among the students living in the on-campus dormitory of a public university located in Kota Kinabalu, Sabah, Malaysia. This study aimed to investigate the outbreak and identify the genotype of the mumps virus (MuV) strain that was involved in the outbreak. MAIN BODY: During the outbreak, one 21-year-old and four 20-year-old males staying in the same dormitory building were reported to have developed symptoms of mumps. Of these, two students were available during the investigation for sample collection to detect MuV by reverse transcription polymerase chain reaction (RT-PCR) of the 639-bp fragment encompassing the entire small hydrophobic (SH) gene. Nucleotide sequencing of the amplicon and phylogenetic analysis using the neighbor-joining method was performed to determine the MuV genotype. Of the two buccal swab samples, one was positive for MuV. The MuV strain in this sample belonged to genotype F and it was clustered together with genotype F strains from China with 96.84-99.68% nucleotide identity. CONCLUSIONS: Genotype F has limited circulation and is endemic in mainland China. Genotype F strains occasionally reported from other countries were epidemiologically linked to China. This study is the first to report a case of genotype F MuV in Malaysia and no epidemiological link could be established with mainland China. The results provide important information that can assist in strategic planning to improve the prevention and control of mumps infection in Malaysia.

High-throughput metabolomics reveals dysregulation of hydrophobic metabolomes in cancer cell lines by Eleusine indica.

Eleusine indica, which is used in traditional medicine, exhibits antiproliferative activity against several cancer cell lines. However, metabolomic studies to evaluate the metabolite changes induced by E. indica in cancer cells are still lacking. The present study investigated the anticancer effects of a root fraction of E. indica (R-S5-C1-H1) on H1299, MCF-7, and SK-HEP-1 cell lines and analyzed metabolic changes in the treated cancer cells using ultra-high-performance liquid chromatography high-resolution mass spectrometry (UHPLC-HRMS). Cell metabolic activity assays demonstrated that the cell viability of the three cancer cell lines was significantly reduced following treatment with R-S5-C1-H1, with half-maximal inhibitory concentrations values of 12.95 µg/mL, 15.99 µg/mL, and 13.69 µg/mL at 72 h, respectively. Microscopy analysis using Hoechst 33342 and Annexin V fluorescent dyes revealed that cells treated with R-S5-C1-H1 underwent apoptotic cell death, while chemometric analysis suggested that apoptosis was triggered 48 h after treatment with R-S5-C1-H1. Deconvoluted cellular metabolomics revealed that hydrophobic metabolites were significantly altered, including triacylglycerols, phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, and ceramide, suggesting that apoptosis induction by R-S5-C1-H1 potentially occurred through modulation of phospholipid synthesis and sphingolipid metabolism. These metabolomic profiling results provide new insights into the anticancer mechanisms of E. indica and facilitate the overall understanding of molecular events following therapeutic interventions.

Carpaine Promotes Proliferation and Repair of H9c2 Cardiomyocytes after Oxidative Insults.

Carpaine has long been identified as the major alkaloid in Carica papaya leaves that possess muscle relaxant properties. Limited study on the molecular signaling properties of carpaine urges us to conduct this study that aims to elucidate the mechanism underlying the cardioprotective effect of carpaine in embryonic cardiomyocytes of the H9c2 cell line. The 50% inhibitory concentration (IC50) of carpaine was first determined using a colorimetric MTT assay to establish the minimum inhibitory concentration for the subsequent test. Using a 1 µM carpaine treatment, a significant increase in the H9c2 proliferation rate was observed following 24 and 48 h of incubation. A Western blot analysis also revealed that carpaine promotes the upregulation of the cell cycle marker proteins cyclin D1 and PCNA. Carpaine-induced H9c2 cell proliferation is mediated by the activation of the FAK-ERK1/2 and FAK-AKT signaling pathways. In the setting of ischemia-reperfusion injury (IRI), carpaine provided a significant protective role to recover the wounded area affected by the hydrogen peroxide (H2O2) treatment. Furthermore, the oxidative-stress-induced reduction in mitochondrial membrane potential (MMP) and overproduction of reactive oxygen species (ROS) were attenuated by carpaine treatment. The current study revealed a novel therapeutic potential of carpaine in promoting in vitro cardiomyocyte proliferation and repair following injury.

A five-year retrospective study on the epidemiology of hand, foot and mouth disease in Sabah, Malaysia.

Hand, foot, and mouth disease (HFMD) is endemic in Malaysia, with the number of cases increasing. Sabah has experienced several HFMD outbreaks, but information on the epidemiology and molecular characteristics of responsible viruses is scarce. In this study, data of 17,574 reports of HFMD cases in Sabah from 2015 to 2019 were extracted from a public health disease surveillance system and analyzed. Twenty-one swab samples from 13 children were collected from Beaufort, Sabah, during an outbreak in August 2018 for detection and serotyping of causative viruses by semi-nested reverse transcription-polymerase chain reaction (snRT-PCR) of the VP4-VP2 region and consensus degenerate hybrid oligonucleotide primer PCR of the VP1 region, respectively. Nucleotide sequencing and phylogenetic analysis were conducted by the neighbor-joining method. The average annual incidence of HFMD was 94.3 per 100,000 people, with the greatest yearly increase between 2017 and 2018. Swabs from six children were tested positive for enterovirus, of which five were positive for CVA16 and one for EV71. All CVA16 strains belonged to sub-genotype B1a, and the EV71 strain belonged to sub-genotype B5. Phylogenetic analyses indicate that enterovirus genotype shift might be responsible for the increasing trend of HFMD in Sabah, however, further study is needed.

Anticancer activity of Momordica cochinchinensis (red gac) aril and the impact of varietal diversity.

BACKGROUND: Momordica cochinchinensis (Cucurbitaceae) is a nutritionally and medicinally important fruit restricted to South East Asia with diverse morphological and genetic variations but there is limited information on its medicinal potential. METHODS: M. cochinchinensis aril from 44 different samples in Australia, Thailand and Vietnam were extracted using different solvents and tested for its anticancer potential. Anticancer activity of M. cochinchinensis aril on breast cancer (MCF7 and BT474) and melanoma (MM418C1 and D24) cells were compared to control fibroblasts (NHDF). The cytotoxicity of the cells following treatment with the aril extract was determined using CCK-8 assay. Biochemical and morphological changes were analysed using flow cytometry, confocal and transmission electron microscopy to determine the mechanism of cell death. RESULTS: The water extract from the aril of M. cochinchinensis elicited significantly higher cytotoxicity towards breast cancer and melanoma cells than the HAE extract. The IC50 concentration for the crude water extract ranged from 0.49 to 0.73 mg/mL and induced both apoptotic and necrotic cell death in a dose- and time-dependant manner with typical biochemical and morphological characteristics. The greatest cytotoxicity was observed from Northern Vietnam samples which caused 70 and 50% melanoma and breast cancer cell death, respectively. CONCLUSIONS: The water extract of M. cochinchinensis aril caused significant apoptosis and necrosis of breast cancer and melanoma cells, with varieties from Northern Vietnam possessing superior activity. This highlights the potential of this fruit in the development of novel anticancer agents against such tumours, with specific regions on where to collect the best variety and extraction solvent for optimum activity.

Extractable impurities from fluoropolymer-based membrane filters - interference in high-throughput, untargeted analysis.

The removal of particles using fluoropolymer-based membrane filters is usually done so to prolong the life span of an analytical column, prevent hardware damage, and reduce signal suppression. Ironically, these membrane filters tend to leach impurities into the samples as the samples are filtered through them. These impurities have the potential to affect the researcher's interpretation in high-throughput, non-targeted analysis. In this study, extractable impurities from different brands of fluoropolymer-based membrane filters present in the filtrate filtered using the said filters were investigated. The results demonstrated that different brand membrane filters and materials tend to elute vastly different numbers of impurities. There were instances whereby the extractable impurities persisted in both the membrane filter and the filtrate despite the filter being pre-conditioned (up to 3 times). Principle component analysis revealed that filtrates at different purge intervals are distant from the unfiltered samples. Pre-conditioning of the PTFE membrane filters could potentially reduce the number of extractable impurities across the tested brands. PVDF filtrates, however, tend to co-cluster with their respective brands, thus suggesting that dissimilarity persists in brands following conditioning. As such, pre-conditioning of the PTFE membrane filters should be encouraged so as to reduce false positive results, while the use of PVDF membrane filters for mass-spectrometry-based untargeted analysis is not advisable as extractable impurities would still persist after 3 rounds of conditioning. Neither the use of different filter brands, nor the use of different filter materials in a sample batch are encouraged as different membrane materials or brands could potentially elute varying impurities.

Norovirus outbreak among students of a boarding school in Kluang, Johor, Malaysia.

INTRODUCTION: Norovirus (NoV) is a contagious virus causing acute gastroenteritis and is mainly responsible for diarrheal outbreak in closed settings. The aims of this study were to describe the epidemiological characteristic of an outbreak in a boarding school, to assess the extent of the outbreak and to implement appropriate control measures. METHODOLOGY: A descriptive study was conducted to describe the epidemiological characteristics of the outbreak. Data on demographic details, onset of abdominal symptoms, food intake history and contact with ill person three days prior to illness were obtained. RESULTS: Twelve fresh stool and 14 food samples were tested for NoV and enteric pathogens, respectively. Out of 745 students, 42 (5.6%) were infected during this outbreak. Predominant clinical features were diarrhea (76.1%), vomiting (71.4%) and abdominal pain (67%). Eight (67%) stool samples and six (43.9%)food samples were positive for NoV and total coliforms, respectively. The dissemination of the disease was due to poor hygiene practices among students. Quarantine was imposed until the last case on September 28, 2016. The outbreak was declared over on September 30, 2016. CONCLUSIONS: A NoV outbreak was determined first time in Malaysia. Environmental assessment showed poor hygienic conditions in the school's kitchen. The number of infected students increased considerably despite the implementation of preventive and control measures. Quarantine was effective to stop the outbreak which is characteristics of NoV outbreak.