Public health expenditure, governance and health outcomes in Malaysia

According to the World Health Organization (WHO), government plays a crucial role in providing quality life for its citizens through good health system. There has been less attention given in analysing the relationship between government expenditure, governance and health outcomes particularly in developing countries. This paper aims to study the impact of public health expenditure and governance on health outcomes in Malaysia. An Autoregressive Distributed Lag (ARDL) cointegration framework has been used to analyse data from 1984 to 2009. The results based on the bounds testing procedure show that a stable, long-run relationship exists between health outcomes and their determinants; namely income level, public health expenditure, corruption and government stability. The results also reveal that public health expenditure and corruption affect long- and short run health outcomes in Malaysia. The findings are important to the policy makers in making decisions to improve the citizens’ quality of life. We suggest the Ministry of Health of Malaysia to conduct more consultations with other ministries and other stakeholders in health services as to identify the needs and emphasize on the importance of health program to the society. At the same time, attention should be given to reduce or eliminate the corruption rate as it has adverse effects on the country

MAA-induced ROS generation in normal human cells.

<p><i>A</i>, Schematic presentation of the pathways predicted to be involved in MAA-induced growth arrest/senescence of normal cells and its recovery by the addition of withanone. <i>B</i>, MAA-induced ROS generation in normal human cells as early as 6–12 h of treatment time. <i>C</i>, ROS level in cells treated with 10 mM MAA for 12 h was double compared to the control. Increase in ROS generation with 5 mM and 10 mM MAA were statistically significant (*P<0.05). <i>D</i>, Cells treated with MAA but recovered in withanone-supplemented medium showed reduction in ROS as compared to the control that were recovered in normal medium. The MAA-induced increase and withanone-induced recovery in ROS generation were statistically significant (*P<0.05).</p

Representative phase contrast images of control, 0.2% or 0,5% ASH-WEX and RA treated cells, in which motility was analyzed by Wound-scratch test (a).

<p>Images show the starting (0 h after scratch) and the end (24 h after scratch) point of the analysis. (b) Graph shows that the rate of IMR-32 migration in response to ASH-WEX treatment in comparison to untreated cells. Data are obtained from a set of scratch test analysis (N  = 3) and are expressed as means ± standard error. Representative MMP zymogram from control and treated samples and their densometery analysis is represented as histogram (c). mRNA expression for MMP2 and MMP9 was analyzed by RT-PCR. Relative percentage expression was expressed as histogram (d). “*” represents the statistical significant (p<0.05) difference between control and ASH-WEX treated groups.</p

MOESM2 of Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence

Additional file 2. Cell lines, history of disease and conditions of incubation throughout the experiments

MOESM3 of Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence

Additional file 3. Schematic presentation of the protocol. A. Determination of standard curve and slope equation. B. QCV assay to determine cell viability, colony forming potential and cell morphology after long-term culture of cells

MOESM1 of Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence

Additional file 1. QCV standardization and determination of slope/y-intercept and R2 value in 16 cell lines

<em>Withania somnifera</em> Water Extract as a Potential Candidate for Differentiation Based Therapy of Human Neuroblastomas

<div><p>Neuroblastoma is an aggressive childhood disease of the sympathetic nervous system. Treatments are often ineffective and have serious side effects. Conventional therapy of neuroblastoma includes the differentiation agents. Unlike chemo-radiotherapy, differentiation therapy shows minimal side effects on normal cells, because normal non-malignant cells are already differentiated. Keeping in view the limited toxicity of <em>Withania somnifera</em> (Ashwagandha), the current study was aimed to investigate the efficacy of Ashwagandha water extract (ASH-WEX) for anti-proliferative potential in neuroblastoma and its underlying signalling mechanisms. ASH-WEX significantly reduced cell proliferation and induced cell differentiation as indicated by morphological changes and NF200 expression in human IMR-32 neuroblastoma cells. The induction of differentiation was accompanied by HSP70 and mortalin induction as well as pancytoplasmic translocation of the mortalin in ASH-WEX treated cells. Furthermore, the ASH-WEX treatment lead to induction of neural cell adhesion molecule (NCAM) expression and reduction in its polysialylation, thus elucidating its anti-migratory potential, which was also supported by downregulation of MMP 2 and 9 activity. ASH-WEX treatment led to cell cycle arrest at G0/G1 phase and increase in early apoptotic population. Modulation of cell cycle marker Cyclin D1, anti-apoptotic marker bcl-xl and Akt-P provide evidence that ASH-WEX may prove to be a promising phytotherapeutic intervention in neuroblatoma related malignancies.</p> </div

Representative Western blots and their densitometry analysis for GFAP (a) and NF200 (e) for RA differentiated C6 and IMR-32 cells, respectively.

<p>RT-PCR results for GFAP and NF200 mRNA in C6 (b) and IMR-32 (f) cells, respectively and their relative densitometry analysis was represented by histograms. The expression of GFAP in C6 (c) and NF200 in IMR-32 (g) cells was analysed by immunocytostaining and relative intensity was plotted as histogram as analysed by Image pro-plus software. “*” represents the statistical significant difference between all the treatment groups (ASH-WEX alone, glutamate alone or glutamate + ASH-WEX groups) with respect to control group. “#” represents the statistical difference between “glutamate + ASH-WEX” treated groups with their respective “glutamate” treatment groups. “*” and “#” = p<0.05.</p

Evaluation and Selection of Candidate Reference Genes for Normalization of Quantitative RT-PCR in <i>Withania somnifera</i> (L.) Dunal

<div><p>Quantitative real-time PCR (qRT-PCR) is now globally used for accurate analysis of transcripts levels in plants. For reliable quantification of transcripts, identification of the best reference genes is a prerequisite in qRT-PCR analysis. Recently, <i>Withania somnifera</i> has attracted lot of attention due to its immense therapeutic potential. At present, biotechnological intervention for the improvement of this plant is being seriously pursued. In this background, it is important to have comprehensive studies on finding suitable reference genes for this high valued medicinal plant. In the present study, 11 candidate genes were evaluated for their expression stability under biotic (fungal disease), abiotic (wounding, salt, drought, heat and cold) stresses, in different plant tissues and in response to various plant growth regulators (methyl jasmonate, salicylic acid, abscisic acid). The data as analyzed by various software packages (geNorm, NormFinder, Bestkeeper and ΔCt method) suggested that cyclophilin (<i>CYP</i>) is a most stable gene under wounding, heat, methyl jasmonate, different tissues and all stress conditions. <i>T-SAND</i> was found to be a best reference gene for salt and salicylic acid (SA) treated samples, while 26S ribosomal RNA (<i>26S</i>), ubiquitin (<i>UBQ</i>) and beta-tubulin (<i>TUB</i>) were the most stably expressed genes under drought, biotic and cold treatment respectively. For abscisic acid (ABA) treated samples <i>18S-rRNA</i> was found to stably expressed gene. Finally, the relative expression level of the three genes involved in the withanolide biosynthetic pathway was detected to validate the selection of reliable reference genes. The present work will significantly contribute to gene analysis studies in <i>W</i>. <i>somnifera</i> and facilitate in improving the quality of gene expression data in this plant as well as and other related plant species.</p></div

MAA-induced loss in mitochondrial membrane potential (MMP) in normal human cells.

<p><i>A</i>, Cells treated with withanone, MAA and MAA→withanone were immunostained for JC1 and mitotracker. MAA-treated cells showed loss of MMP (loss of red staining). MAA→withanone treated cells showed mitochondrial membrane potential comparable to that of the control cells. <i>B</i>, Quantitation of cells showing red JC1 staining indicative of high mitochondrial membrane potential. MAA treatment caused decrease in the number of cells with high MMP and the recovery in the withanone-supplemented medium resulted in an increase in the number of cells with high MMP (*P<0.05). <i>C</i>, Cell viability analysis of the control, H₂O₂ and H₂O₂→withanone treated cells showing the recovery of cells. Cell viability was increased (statistical significance, *P<0.05) when they were recovered from the oxidative stress by incubation in the withanone-supplemented medium.</p