Determination toxic effects of Hystrix Brachyura Bezoar extracts using cancer cell lines and embryo zebrafish (Danio rerio) models and identification of active principles through GC-MS analysis

Ethnopharmacological relevance: Porcupine bezoar (PB) is used as folk medicine for various medical conditions including cancer treatment in Malaysia. However, its toxicity profile has never been thoroughly ascertained to confirm its safe nature as an efficacious traditional medicine in the treatment of cancer as well as other ailments. Aim of the study: This study was aimed to reveal three different PBs’ aqueous extracts(viz. PB-A, PB-B, PB-C) chemical constituent’s profile using GC-MS analysis, anticancer property on A375, HeLa and MCF7 cancer cells, toxicity profile on zebrafish embryo morphology, EC50, LC50 and teratogenicity index. Materials and methods: PBs’ extracts characterization was performed through GC-MS analysis, in vitro anticancer effect was carried out on A375, HeLa and MCF7 cancer cell lines and finally and toxicity properties on three different PBs aqueous extracts (viz. PB-A, PB-B, PB-C) were determined using zebrafish embryo model. Results: The GC-MS analysis revealed 10 similar compounds in all PBs’ extracts. Dilauryl thiodipropionate was found to be a major compound in all PBs’ extracts followed by tetradecanoic acid. An in vitro anticancer study revealed PB extracts exerted median inhibition concentration (IC50) <50 μg/mL, on cancer cells viz. A375, HeLa and MCF7 with no significant toxicity on normal cells viz. NHDF cells. In vivo toxicity of PBs extracts found affecting tail detachment, hatching, craniofacial, brain morphology, soft tissues, edema, spinal, somites, notochord and cardiovascular system (brachycardia, disruption of blood circulation) deformities. The LC50 and EC50 demonstrated PB extracts effect as dose and time dependent with median concentration <150.0 μg/mL. Additionally, teratogenicity index (TI) viz. >1.0 revealed teratogenic property for PB extracts. Conclusions: The findings revealed that all three PBs aqueous extracts possessed anticancer activity and exhibited significant toxicological effects on zebrafish embryos with high teratogenicity index. Hence, its use as an anticancer agent requires further investigation and medical attentions to determine its safe dose

Mapping geographical inequalities in access to drinking water and sanitation facilities in low-income and middle-income countries, 2000-17

Background: Universal access to safe drinking water and sanitation facilities is an essential human right, recognised in the Sustainable Development Goals as crucial for preventing disease and improving human wellbeing. Comprehensive, high-resolution estimates are important to inform progress towards achieving this goal. We aimed to produce high-resolution geospatial estimates of access to drinking water and sanitation facilities. Methods: We used a Bayesian geostatistical model and data from 600 sources across more than 88 low-income and middle-income countries (LMICs) to estimate access to drinking water and sanitation facilities on continuous continent-wide surfaces from 2000 to 2017, and aggregated results to policy-relevant administrative units. We estimated mutually exclusive and collectively exhaustive subcategories of facilities for drinking water (piped water on or off premises, other improved facilities, unimproved, and surface water) and sanitation facilities (septic or sewer sanitation, other improved, unimproved, and open defecation) with use of ordinal regression. We also estimated the number of diarrhoeal deaths in children younger than 5 years attributed to unsafe facilities and estimated deaths that were averted by increased access to safe facilities in 2017, and analysed geographical inequality in access within LMICs. Findings: Across LMICs, access to both piped water and improved water overall increased between 2000 and 2017, with progress varying spatially. For piped water, the safest water facility type, access increased from 40·0% (95% uncertainty interval [UI] 39·4–40·7) to 50·3% (50·0–50·5), but was lowest in sub-Saharan Africa, where access to piped water was mostly concentrated in urban centres. Access to both sewer or septic sanitation and improved sanitation overall also increased across all LMICs during the study period. For sewer or septic sanitation, access was 46·3% (95% UI 46·1–46·5) in 2017, compared with 28·7% (28·5–29·0) in 2000. Although some units improved access to the safest drinking water or sanitation facilities since 2000, a large absolute number of people continued to not have access in several units with high access to such facilities (>80%) in 2017. More than 253 000 people did not have access to sewer or septic sanitation facilities in the city of Harare, Zimbabwe, despite 88·6% (95% UI 87·2–89·7) access overall. Many units were able to transition from the least safe facilities in 2000 to safe facilities by 2017; for units in which populations primarily practised open defecation in 2000, 686 (95% UI 664–711) of the 1830 (1797–1863) units transitioned to the use of improved sanitation. Geographical disparities in access to improved water across units decreased in 76·1% (95% UI 71·6–80·7) of countries from 2000 to 2017, and in 53·9% (50·6–59·6) of countries for access to improved sanitation, but remained evident subnationally in most countries in 2017. Interpretation: Our estimates, combined with geospatial trends in diarrhoeal burden, identify where efforts to increase access to safe drinking water and sanitation facilities are most needed. By highlighting areas with successful approaches or in need of targeted interventions, our estimates can enable precision public health to effectively progress towards universal access to safe water and sanitation

Prevalence and attributable health burden of chronic respiratory diseases, 1990–2017 : a systematic analysis for the Global Burden of Disease Study 2017

Background Previous attempts to characterise the burden of chronic respiratory diseases have focused only on specific disease conditions, such as chronic obstructive pulmonary disease (COPD) or asthma. In this study, we aimed to characterise the burden of chronic respiratory diseases globally, providing a comprehensive and up-to-date analysis on geographical and time trends from 1990 to 2017. Methods Using data from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2017, we estimated the prevalence, morbidity, and mortality attributable to chronic respiratory diseases through an analysis of deaths, disability-adjusted life-years (DALYs), and years of life lost (YLL) by GBD super-region, from 1990 to 2017, stratified by age and sex. Specific diseases analysed included asthma, COPD, interstitial lung disease and pulmonary sarcoidosis, pneumoconiosis, and other chronic respiratory diseases. We also assessed the contribution of risk factors (smoking, second-hand smoke, ambient particulate matter and ozone pollution, household air pollution from solid fuels, and occupational risks) to chronic respiratory disease-attributable DALYs. Findings In 2017, 544.9 million people (95% uncertainty interval [UI] 506.9- 584.8) worldwide had a chronic respiratory disease, representing an increase of 39.8% compared with 1990. Chronic respiratory disease prevalence showed wide variability across GBD super-regions, with the highest prevalence among both males and females in high-income regions, and the lowest prevalence in sub-Saharan Africa and south Asia. The age-sex- specific prevalence of each chronic respiratory disease in 2017 was also highly variable geographically. Chronic respiratory diseases were the third leading cause of death in 2017 (7.0% [95% UI 6.8-7 .2] of all deaths), behind cardiovascular diseases and neoplasms. Deaths due to chronic respiratory diseases numbered 3 914 196 (95% UI 3 790 578-4 044 819) in 2017, an increase of 18.0% since 1990, while total DALYs increased by 13.3%. However, when accounting for ageing and population growth, declines were observed in age-standardised prevalence (14.3% decrease), agestandardised death rates (42.6%), and age-standardised DALY rates (38.2%). In males and females, most chronic respiratory disease-attributable deaths and DALYs were due to COPD. In regional analyses, mortality rates from chronic respiratory diseases were greatest in south Asia and lowest in sub-Saharan Africa, also across both sexes. Notably, although absolute prevalence was lower in south Asia than in most other super-regions, YLLs due to chronic respiratory diseases across the subcontinent were the highest in the world. Death rates due to interstitial lung disease and pulmonary sarcoidosis were greater than those due to pneumoconiosis in all super-regions. Smoking was the leading risk factor for chronic respiratory disease-related disability across all regions for men. Among women, household air pollution from solid fuels was the predominant risk factor for chronic respiratory diseases in south Asia and sub-Saharan Africa, while ambient particulate matter represented the leading risk factor in southeast Asia, east Asia, and Oceania, and in the Middle East and north Africa super-region. Interpretation Our study shows that chronic respiratory diseases remain a leading cause of death and disability worldwide, with growth in absolute numbers but sharp declines in several age-standardised estimators since 1990. Premature mortality from chronic respiratory diseases seems to be highest in regions with less-resourced health systems on a per-capita basis

Prevalence and attributable health burden of chronic respiratory diseases, 1990–2017: A systematic analysis for the global burden of disease study 2017

© 2020 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license Background: Previous attempts to characterise the burden of chronic respiratory diseases have focused only on specific disease conditions, such as chronic obstructive pulmonary disease (COPD) or asthma. In this study, we aimed to characterise the burden of chronic respiratory diseases globally, providing a comprehensive and up-to-date analysis on geographical and time trends from 1990 to 2017. Methods: Using data from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2017, we estimated the prevalence, morbidity, and mortality attributable to chronic respiratory diseases through an analysis of deaths, disability-adjusted life-years (DALYs), and years of life lost (YLL) by GBD super-region, from 1990 to 2017, stratified by age and sex. Specific diseases analysed included asthma, COPD, interstitial lung disease and pulmonary sarcoidosis, pneumoconiosis, and other chronic respiratory diseases. We also assessed the contribution of risk factors (smoking, second-hand smoke, ambient particulate matter and ozone pollution, household air pollution from solid fuels, and occupational risks) to chronic respiratory disease-attributable DALYs. Findings: In 2017, 544·9 million people (95% uncertainty interval [UI] 506·9–584·8) worldwide had a chronic respiratory disease, representing an increase of 39·8% compared with 1990. Chronic respiratory disease prevalence showed wide variability across GBD super-regions, with the highest prevalence among both males and females in high-income regions, and the lowest prevalence in sub-Saharan Africa and south Asia. The age-sex-specific prevalence of each chronic respiratory disease in 2017 was also highly variable geographically. Chronic respiratory diseases were the third leading cause of death in 2017 (7·0% [95% UI 6·8–7·2] of all deaths), behind cardiovascular diseases and neoplasms. Deaths due to chronic respiratory diseases numbered 3 914 196 (95% UI 3 790 578–4 044 819) in 2017, an increase of 18·0% since 1990, while total DALYs increased by 13·3%. However, when accounting for ageing and population growth, declines were observed in age-standardised prevalence (14·3% decrease), age-standardised death rates (42·6%), and age-standardised DALY rates (38·2%). In males and females, most chronic respiratory disease-attributable deaths and DALYs were due to COPD. In regional analyses, mortality rates from chronic respiratory diseases were greatest in south Asia and lowest in sub-Saharan Africa, also across both sexes. Notably, although absolute prevalence was lower in south Asia than in most other super-regions, YLLs due to chronic respiratory diseases across the subcontinent were the highest in the world. Death rates due to interstitial lung disease and pulmonary sarcoidosis were greater than those due to pneumoconiosis in all super-regions. Smoking was the leading risk factor for chronic respiratory disease-related disability across all regions for men. Among women, household air pollution from solid fuels was the predominant risk factor for chronic respiratory diseases in south Asia and sub-Saharan Africa, while ambient particulate matter represented the leading risk factor in southeast Asia, east Asia, and Oceania, and in the Middle East and north Africa super-region. Interpretation: Our study shows that chronic respiratory diseases remain a leading cause of death and disability worldwide, with growth in absolute numbers but sharp declines in several age-standardised estimators since 1990. Premature mortality from chronic respiratory diseases seems to be highest in regions with less-resourced health systems on a per-capita basis. Funding: Bill & Melinda Gates Foundation

Tracking development assistance for health and for COVID-19: a review of development assistance, government, out-of-pocket, and other private spending on health for 204 countries and territories, 1990-2050

Background The rapid spread of COVID-19 renewed the focus on how health systems across the globe are financed, especially during public health emergencies. Development assistance is an important source of health financing in many low-income countries, yet little is known about how much of this funding was disbursed for COVID-19. We aimed to put development assistance for health for COVID-19 in the context of broader trends in global health financing, and to estimate total health spending from 1995 to 2050 and development assistance for COVID-19 in 2020. Methods We estimated domestic health spending and development assistance for health to generate total health-sector spending estimates for 204 countries and territories. We leveraged data from the WHO Global Health Expenditure Database to produce estimates of domestic health spending. To generate estimates for development assistance for health, we relied on project-level disbursement data from the major international development agencies' online databases and annual financial statements and reports for information on income sources. To adjust our estimates for 2020 to include disbursements related to COVID-19, we extracted project data on commitments and disbursements from a broader set of databases (because not all of the data sources used to estimate the historical series extend to 2020), including the UN Office of Humanitarian Assistance Financial Tracking Service and the International Aid Transparency Initiative. We reported all the historic and future spending estimates in inflation-adjusted 2020 US$, 2020 US$ per capita, purchasing-power parity-adjusted US$per capita, and as a proportion of gross domestic product. We used various models to generate future health spending to 2050. Findings In 2019, health spending globally reached$8. 8 trillion (95% uncertainty interval UI] 8.7-8.8) or $1132 (1119-1143) per person. Spending on health varied within and across income groups and geographical regions. Of this total,$40.4 billion (0.5%, 95% UI 0.5-0.5) was development assistance for health provided to low-income and middle-income countries, which made up 24.6% (UI 24.0-25.1) of total spending in low-income countries. We estimate that $54.8 billion in development assistance for health was disbursed in 2020. Of this,$13.7 billion was targeted toward the COVID-19 health response. $12.3 billion was newly committed and$1.4 billion was repurposed from existing health projects. $3.1 billion (22.4$2.4 billion (17.9%) was for supply chain and logistics. Only $714.4 million (7.7$1519 (1448-1591) per person in 2050, although spending across countries is expected to remain varied. Interpretation Global health spending is expected to continue to grow, but remain unequally distributed between countries. We estimate that development organisations substantially increased the amount of development assistance for health provided in 2020. Continued efforts are needed to raise sufficient resources to mitigate the pandemic for the most vulnerable, and to help curtail the pandemic for all. Copyright (C) 2021 The Author(s). Published by Elsevier Ltd

Cancer chemoprevention study of Luffa Aegyptiaca seed extract on human breast cancer cell lines (MCF-7)

Objectives/Research Problem: Breast cancer is a major health problem in Malaysia and the world. It is the commonest malignancy in women and the second leading cause of cancer deaths. There are many conventional treatments available for breast cancer patients, such as chemotherapy, radiation therapy, immune therapy, hormone therapy and many more. However, drug resistance acquired by cancer cells has led to treatment failure. Alternative treatment with minimum or no side effect is highly demanded. Therefore, the aim of this study was to determine the chemopreventive effect of Luffa aegyptiaca seed extract (LSE) on breast cancer through anti-proliferative effect and its mechanisms on breast cancer cell (MCF-7). It can be evaluated by result of IC₅₀ determination, cell proliferation assay and flow cytometer analysis on apoptosis induction and cell cycle arrest. Materials and Method: MCF-7 cells were seeded at a density of 4.7 x 10⁴ cells/well. Trypan blue exclusion method was used to determine the viable cells using haemocytometer. The IC50 assay was determined against MCF-7 cell at different concentrations of LSE administration. The morphological changes of MCF-7 cells also were observed. MCF-7 cell and 3T3-L1 cell were treated with LSE at IC50 to evaluate the cell proliferation assay and cytotoxicity assay, respectively. The induction of apoptosis and alteration on cell cycle regulation will be assessed by flow cytometer. Results and Discussion: LSE showed the growth inhibitory effect with IC₅₀ value at 0.0625 mg/ml after 72 hours exposure. Morphological changes displaying apoptosis also had been observed upon treatment with LSE at IC50. Moreover, LSE showed no toxicity effect on normal cells (3T3-L1). Thus proved the potential chemopreventive agent of the extracts on MCF-7. The results for flow cytometric analysis are yet to be determined as the works are still in progress. Conclusion: Overall, the data collected provided new insight of using the Luffa aegyptiaca seed which can be used as chemopreventive agents on MCF-7. KEYWORDS: Cancer Chemoprevention, Breast Cancer, Luffa aegyptiaca, Anti-Proliferative Effect, IC₅₀ Determination, Apoptosis, Cell Cycle, MCF-

Cancer chemoprevention study of luffa aegyptiaca (sponge gourd) seed extract on human breast cancer cell lines (MCF-7)

Natural products from plant have been used for centuries to prevent various diseases including cancer. Many chemopreventive agents are believed to inhibit or delay the progression of cancer cells through suppression of cell proliferation, induction of apoptosis and cell cycle arrest. In Malaysia, breast cancer is the most common form of cancer affecting women. The prognosis for breast cancer is still poor and conventional treatment is not always effective. Therefore, this study aims to determine the chemopreventive effect of Luffa aegyptiaca seed extract (LSE) on breast cancer through anti-proliferative effect and its mechanisms on breast cancer cell (MCF-7) as evidenced by the result of IC50 determination, cell proliferation assay and flow cytometer analysis on apoptosis induction and cell cycle arrest. MCF-7 cells were plated at a density of 4.7 x 104 cells/ml in 6–well plate. Trypan blue exclusion method was used to determine cell viability. Untreated and LSE-treated MCF-7 cells were observed for morphological changes with an inverted-phase contrast microscope. The induction of apoptosis and cell cycle arrest was analysed using flow cytometry. LSE showed growth inhibitory effect with IC50 value at 0.0625 mg/ml. Moreover, LSE, show no toxic effect on normal cells possibly showing that it is a potential chemopreventive agent on MCF-7 via the induction of apoptosis and cell cycle arrest. Overall, the data collected provides new insight for the use of Luffa aegyptiaca seed as a chmopreventive agent on MCF-7. Keywords: Luffa aegyptiaca; anti-proliferative effect; apoptosis; cell cycle; MCF-

Understanding of anti-cancer properties of neolamarckia cadamba leaves extract on breast cancer cell

Objectives/Research Problem: The increasing of breast cancer cases from year to year is really worrying. It is ranked as the most common cancer occurs and the leading cause of cancer death among women worldwide. Although there are many advance treatments available, the long term effect of the treatments on the patients cannot be ensured. Effective, safe and non-toxic treatment of natural product is one of the best approaches in treating breast cancer and act as chemopreventive agent. Neolamarckia cadamba, an Ayurvedic medicinal plant, has been used by Indian folklore in treating illnesses. The variety of phytochemical compounds found and pharmacological activities in this Rubiaceae family plant indicates that it has a strong potential in treating cancer. Therefore, there is a need to evaluate anti-cancer properties of this plant on breast cancer cell. The present study is carried out to obtain a better understanding on chemopreventive effect of N. cadamba’s leaves on human breast cancer cell lines (MCF-7). Materials and Method: N. cadamba leaves were extracted with 80% ethanol by sonication method. Different concentrations of N. cadamba leaves extract (NCE) were prepared by serial dilution (1:2). Then, the extracts were administered on breast cancer cell line (MCF-7). The 50% inhibitory concentration (IC50) assay was conducted by Trypan blue exclusion method to determine the cell viability. The IC50 value will be used to measure antiproliferative effect. The induction of apoptosis and cell cycle arrest will be evaluated using flow cytometry. The mechanism pathway which regard to apoptosis and cell cycle will be determined using Quantitative Polymerase Chain Reaction (qPCR). Results and Discussion: The result found that the IC50 value of this extract was 0.125 mg/ml. It showed that the NCE have ability to inhibit growth of breast cancer cell. Meanwhile, the results for antiproliferative assay, flow cytometric analysis and qPCR analysis are yet to be determined as the works are still in progress. Conclusion: The present study demonstrates that the N. cadamba leaves extract has a potential against breast cancer cell line, MCF-7. KEYWORDS: Neolamarckia. cadamba, Leaves Extract, IC50, Breast Cance

<i>Hystrix Brachyura</i> Bezoar Characterization, Antioxidant Activity Screening, and Anticancer Activity on Melanoma Cells (A375): A Preliminary Study

Porcupine bezoars (PBs) are masses of undigested calcareous concretions formed within the gastrointestinal tract. There are undocumented claims that PBs have antioxidant activity and can treat cancers. However, limited scientific study has been carried out to verify these traditional claims. Hence, this study was conducted to characterize the chemical profile and validate the antioxidant and anticancer activity against melanoma cells (A375). PB extract was initially subjected to Fourier-transform infrared spectroscopy (FTIR), gas chromatography&#8315;mass spectrometry (GCMS), total phenolic content (TPC), and total flavonoid content (TFC) analyses. The bioautography of antioxidant assays, namely 2,2&#8242;-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazy (DPPH), and &#946;-carotene was performed. An in vitro A375 cell viability assay, apoptosis assay, cell cycle arrest assay, and gene expression assay were carried out as well. The experimental finding revealed 5,10-diethoxy-2,3,7,8-tetrahydro-1H,6H-dipyrrolo[1,2-a:1&#8242;,2&#8242;-d]pyrazine, ursodeoxycholic acid, and cholest-5-en-3-ol (3 beta)-, carbonochloridate are major compounds detected in PB extract. PB extract has low phenolic content, viz. 698.7 &#177; 0.93 (&#181;g GAE/5 mg dry weight). The bioautography antioxidant assays revealed a potent antioxidant effect (ABTS &gt; DPPH &gt; &#946;-carotene), with free radical scavenging activity. Furthermore, PB extract exhibited dose- and time-dependent inhibition of cancer activity on A375 cells due to the exhibition of apoptosis via an intrinsic pathway

Understanding of anti-cancer properties of Neolamarckia cadamba (Kalempayan) leaves extract on breast cancer cell

Natural products have been used in treating illnesses since ancient times. Neolamarckia cadamba (N. cadamba) plant, locally known as Kalempayan, is an Ayurvedic medicine. It has been used by Indian folklore for treatment such as fever, anaemia, stomatitis, and infectious diseases. This Rubiaceae family plant indicates that it has a therapeutic potential against diseases, including cancer. Therefore, this study aimed to investigate the chemopreventive effect of N. cadamba’s leaves extract (NCE) with regard to the induction of apoptosis and cell cycle arrest against human breast adenocarcinoma cancer cell-line (MCF-7). MCF-7 cells were seeded at 4.7 x 104 cells/well in 6-well plates. The cells were treated with series concentration of NCE. The 50% inhibitory concentration (IC50) of NCE and its antiproliferative effect were measured using Trypan blue exclusion method was determined as 0.125 mg/ml. The flow cytometric analysis was performed in MCF-7 revealed the NCE inhibited the cell proliferation through induction of apoptosis and cell cycle arrest. The present study demonstrates that the NCE has a positive potential as a chemopreventive agent. Further research need to be done to identify the molecular mechanism related to induction of apoptosis and cell cycle arrest in breast cancer cells