Curcumin piperidone derivatives induce caspase-dependent apoptosis and suppress miRNA-21 expression in LN-18 human glioblastoma cells.

BACKGROUND: Previously, we have reported on the two curcuminoid analogues with piperidone derivatives, namely FLDP-5 and FLDP-8 have more potent anti-proliferative and anti-migration effects than curcumin. In this study, we further investigated the mode of cell death and the mechanism involved in the cell death process induced by these analogues on human glioblastoma LN-18 cells. RESULTS: The FLDP-5 and FLDP-8 curcuminoid analogues induced LN-18 cell death through apoptosis in a concentration-dependent manner following 24 h of treatment. These analogues induced apoptosis in LN-18 cells through significant loss of mitochondrial mass and mitochondrial membrane potential (MMP) as early as 1-hour of treatment. Interestingly, N-acetyl-l-cysteine (NAC) pretreatment did not abolish the apoptosis induced by these analogues, further confirming the cell death process is independent of ROS. However, the apoptosis induced by the analogues is caspases-dependent, whereby pan-caspase pretreatment inhibited the curcuminoid analogues-induced apoptosis. The apoptotic cell death progressed with the activation of both caspase-8 and caspase-9, which eventually led to the activation of caspase-3, as confirmed by immunoblotting. Moreover, the existing over-expression of miRNA-21 in LN-18 cells was suppressed following treatment with both analogues, which suggested the down-regulation of the miRNA-21 facilitates the cell death process. CONCLUSION: The FLDP-5 and FLDP-8 curcuminoid analogues downregulate the miRNA-21 expression and induce extrinsic and intrinsic apoptotic pathways in LN-18 cells.

Precision Medicine: Making It Happen for Malaysia.

Precision medicine is transforming healthcare worldwide and aims to improve the effectiveness of management of many diseases including cancers, other non-communicable diseases (NCDs) and also rare diseases. Precision medicine takes into account the individual patient's genetic, environment and lifestyle data. Developed nations are already embarking on precision medicine initiatives including the 100,000 Genomes England and the Precision Medicine Initiative in the United States (US). The Academy of Sciences Malaysia, the Ministry of Health and the Ministry of Higher Education are working together to put forward a precision medicine initiative for Malaysia. The key drivers that must be put in place include a strong policy agenda, a national large scale genome sequencing project and with it a national genome database, the implementation of the electronic medical record (EMR) system, a payment and reimbursement system to cover for the genetic testing and the targeted treatment, and putting in place an ecosystem that will support precision medicine. Relevant guidelines and Acts will also need to be developed especially with regard to privacy and confidentiality. The future of precision medicine is now and this will certainly bring better outcome and value to the patients.

Prevalence and Histopathological Characteristics of KCNJ5 Mutant Aldosterone-Producing Adenomas in a Multi-Ethnic Malaysian Cohort.

Studies on excised adrenals from primary aldosteronism patients have found that somatic mutations in KCNJ5 frequently cause excess aldosterone production in the culprit aldosterone-producing adenoma (APA). KCNJ5 mutant APAs were reported to be peculiarly overrepresented among young females and in Oriental cohorts, compared to their older male, or Caucasian counterparts. These larger APAs were also reported to have similarities with the zona fasciculata (ZF) in the adrenal both from the steroid production profile and the morphology of the cell. We therefore aimed to corroborate these findings by characterizing the APAs from a multi-ethnic Malaysian cohort. The prevalence of KCNJ5 mutations was estimated through targeted DNA sequencing of KCNJ5 in 54 APAs. Confirmation of APA sample acquisition was performed by CYP11B2 immunohistochemistry (IHC) staining. The ZF steroid production profile was based on the ZF enzyme CYP17A1 IHC staining, and ZF cell morphology was based on a high cytoplasm to nucleus ratio. Seventeen (31.5%) APAs studied, harbored a KCNJ5 mutation. No female over-representation was seen in this cohort though females were found to have a higher expression of CYP11B2 than males (p = 0.009; Mann-Whitney U test). Age at adrenalectomy correlated negatively with the percentage of ZF-like cells in the APA (p = 0.01; Spearman's rho) but not with the KCNJ5 genotype. KCNJ5 mutant APAs had a high percentage of ZF-like cells (and high CYP17A1 expression) but so did the wild-type APAs. In summary, prevalence of KCNJ5 mutant APAs in this cohort was similar to other Caucasian cohorts, however, over-representation of females did not occur, which is similar to some studies in Oriental cohorts.

First reported case of Haemoglobin-M Hyde Park in a Malay family living in Malaysia.

Haemoglobin (Hb)-M Hyde Park, also known as Hb-M Akita is a rare type of hereditary Hb M due to autosomal dominant mutation of CAC>TAC on codon 92 of ß globin gene resulting in the replacement of histidine by tyrosine on ß globin chain. This variant Hb has a tendency to form methaemoglobin (metHb). The iron ion in metHb is oxidized to ferric (Fe3+) which is unable to carry oxygen and the patients manifest as cyanosis clinically. A 9-year-old Malay girl was incidentally found to be cyanotic when she presented to a health clinic. Laboratory investigations revealed raised methaemoglobin levels and Hb analysis findings were consistent with Hb-M Hyde Park. ß gene sequencing confirmed a point mutation of CAC>TAC on codon 92 in one of the ß genes. The family study done on the individuals with cyanosis showed similar findings. A diagnosis of heterozygous Hb-M Hyde Park was made. Patients with this variant Hb usually presented with cyanosis with mild haemolysis and maybe misdiagnosed as congenital heart disease. No further treatment is needed as patients are relatively asymptomatic. Although the disease is harmless in the heterozygous carriers but the offspring of the carriers may suffer severe haemolytic anaemia when the offspring also inherit other ß haemoglobinopathies/thalassemia. This can happen due to high prevalence of ß thalassemia carrier (3.5-4 %) found in Malaysia. At the time of writing, this is the first case of hereditary Hb-M Hyde Park diagnosed in a Malay family living in Malaysia.

Gamma-tocotrienol and hydroxy-chavicol synergistically inhibits growth and induces apoptosis of human glioma cells.

BACKGROUND: Gamma-tocotrienol (GTT), an isomer of vitamin E and hydroxy-chavicol (HC), a major bioactive compound in Piper betle, has been reported to possess anti-carcinogenic properties by modulating different cellular signaling events. One possible strategy to overcome multi-drug resistance and high toxic doses of treatment is by applying combinational therapy especially using natural bioactives in cancer treatment. METHODS: In this study, we investigated the interaction of GTT and HC and its mode of cell death on glioma cell lines. GTT or HC alone and in combination were tested for cytotoxicity on glioma cell lines 1321N1 (Grade II), SW1783 (Grade III) and LN18 (Grade IV) by [3-(4,5-dimethylthiazol-2- yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)- 2H- tetrazolium, inner salt] MTS assay. The interactions of each combination were evaluated by using the combination index (CI) obtained from an isobologram. RESULTS: Individually, GTT or HC displayed mild growth inhibitory effects against glioma cancer cell lines at concentration values ranging from 42-100 µg/ml and 75-119 µg/ml respectively. However, the combination of sub-lethal doses of GTT + HC dramatically enhanced the inhibition of glioma cancer cell proliferation and exhibited a strong synergistic effect on 1321N1 with CI of 0.55, and CI = 0.54 for SW1783. While in LN18 cells, moderate synergistic interaction of GTT + HC was observed with CI value of 0.73. Exposure of grade II, III and IV cells to combined treatments for 24 hours led to increased apoptosis as determined by annexin-V FITC/PI staining and caspase-3 apoptosis assay, showing caspase-3 activation of 27%, 7.1% and 79% respectively. CONCLUSION: In conclusion, combined treatments with sub-effective doses of GTT and HC resulted in synergistic inhibition of cell proliferation through the induction of apoptosis of human glioma cells in vitro.

The state of emergency care in the Republic of the Sudan

Sudan is one of the largest African countries; covering an area of 1.9million km2-approximately one fifth of the geographic area of the United States. The population is 30million people; the majority of whom (68) live in rural areas; as compared with the sub-Saharan African average of approximately 62. Sudan is considered a lower-middle income country-with 47 of the population living below the poverty line and a gross domestic product (GDP) of US $62billion in 2010. In addition to excessive burden of communicable diseases such as malaria; tuberculosis; and schistosomiasis; Sudan is particularly susceptible to both natural and manmade disasters. Drought and flood are quite common due to Sudan's proximity to and dependency on the Nile; and throughout history Sudan has also been plagued with internal conflicts and outbreaks of violence; which bring about a burden of traumatic disease and demand high quality emergency care. The purpose of this paper is to describe the state of emergency care and Emergency Medicine education; and their context within the Sudanese health care system. As is the case in most African countries; emergency care is delivered by junior staff: new graduates from medical schools and unsupervised medical officers who handle all types of case presentations. In 2001; increased mortality and morbidity among unsorted patients prompted the Ministry of Health to introduce a new triage-based emergency care system. In late 2005; twenty-one Emergency physicians delivered these new Emergency Services. In 2011; following a curriculum workshop in November 2010; the Emergency Medicine residency program was started in Khartoum. Currently there are 27 rotating registrars; the first class of whom is expected to graduate in 2015

Inhibition of beta-catenin and KRAS expressions by Piper betle in azoxymethane-induced colon cancer of male Fischer 344 rats.

OBJECTIVE: To investigate the chemopreventive effect of Piper betle (PB) on preneoplastic lesions (aberrant crypt foci [ACF]) induced by azoxymethane (AOM) in rats and its effect on colorectal cancer biomarkers (beta-catenin, KRAS, p53 and p21). STUDY DESIGN: A total of 32 male Fischer 344 rats were divided into phase 1 and phase 2 groups (8 and 24 weeks of AOM administration, respectively). Each phase was divided into 4 groups: control or normal saline (NS) (1 mL/kg), AOM (15 mg/kg body weight, once weekly for 2 weeks), PB (75 mg/kg body weight) and AOM + PB. PB was force-fed to rats a week after the second dose of AOM and NS. The colon was cut open longitudinally for methylene blue and immunohistochemistry staining. RESULTS: AOM administration showed formation of ACF at 8 and 24 weeks. PB, however, did not reduce ACF formation at either week, but it managed to reduce beta-catenin expression and KRAS found highly expressed in the AOM group of phase 1 rats. No immunoreactivities of p53 and p21 were detected in phase 2 rats, but instead inflammatory cells were visible in between the lesions. CONCLUSION: PB may act as a potential chemopreventive agent in the early stage of colon carcinogenesis by suppressing the expressions of beta-catenin and KRAS.

Fish oil supplementation is beneficial on caloric intake, appetite and mid upper arm muscle circumference in children with leukaemia.

A randomised trial was carried out to determine the effect of supplementation of fish oil among 51 children with leukaemia aged 4 to 12 years on appetite level, caloric intake, body weight and lean body mass. They were randomly allocated into the trial group (TG) and the control group (CG). At baseline, 30.8% of TG subjects and 44.0% of CG subjects were malnourished and 7.7% of subject from TG and 28.0% from CG were classified as stunted. The majority of subjects from TG and CG were in the mild malnutrition category for mid upper arm muscle circumference (MUAMC)-for-age. The TG group showed significant increment in MUAMC (0.13 cm vs -0.09 cm) compared with CG at 8 weeks (p<0.001). There was a significant higher increase for appetite level (0.12±0.33) (p<0.05) and an increasing trend on energy and protein intake in the TG group (213±554 kcal; 3.64 ±26.8 g) than in the CG group. In conclusion, supplementation of fish oil has a positive effect on appetite level, caloric intake and MUAMC among children with leukaemia.

Gene expression profiling and cancer-related pathways in type I endometrial carcinoma.

INTRODUCTION: Malignant transformation of type I endometrium involves alteration in gene expression with subsequent uncontrolled proliferation of altered cells. OBJECTIVE: The main objective of the present study was to identify the cancer-related genes and gene pathways in the endometrium of healthy and cancer patients. MATERIALS AND METHODS: Thirty endometrial tissues from healthy and type I EC patients were subjected to total RNA isolation. The RNA samples with good integrity number were hybridized to a new version of Affymetrix Human Genome GeneChip 1.0 ST array. We analyzed the results using the GeneSpring 9.0 GX and the Pathway Studio 6.1 software. For validation assay, quantitative real-time polymerase chain reaction was used to analyze 4 selected genes in normal and EC tissue. RESULTS: Of the 28,869 genes profiled, we identified 621 differentially expressed genes (2-fold) in the normal tissue and the tumor. Among these genes, 146 were up-regulated and 476 were down-regulated in the tumor as compared with the normal tissue (P < 0.001). Up-regulated genes included the v-erb-a erythroblastic leukemia viral oncogene homolog 3 (ErbB3), ErbB4, E74-like factor 3 (ELF3), and chemokine ligand 17 (CXCL17). The down-regulated genes included signal transducer and activator transcription 5B (STAT5b), transforming growth factor A receptor III (TGFA3), caveolin 1 (CAV1), and protein kinase C alpha (PKCA). The gene set enrichment analysis showed 10 significant gene sets with related genes (P < 0.05). The quantitative polymerase chain reaction of 4 selected genes using similar RNA confirmed the microarray results (P < 0.05). CONCLUSIONS: Identification of molecular pathways with their genes related to type I EC contribute to the understanding of pathophysiology of this cancer, probably leading to identifying potential biomarkers of the cancer.

Preimplantation genetic diagnosis for beta-thalassemia using single-cell DNA analysis for codons 17 and 26 of beta-globin gene.

BACKGROUND: Preimplantation genetic diagnosis (PGD) of monogenic autosomal hereditary disorders following assisted conception usually involves the removal of one or two blastomeres from preimplantation embryos. However, the amount of DNA from a single blastomere is insufficient to amplify the region of interest. Hence, the whole genome amplification (WGA) method is performed prior to amplifying the genes of interest before analysis of DNA material through polymerase chain reaction (PCR). METHODS: In the present study we report that WGA from a single blastomere extracted from unwanted preimplantation human embryos (obtained from 10 infertile couples) could positively yield microgram quantities of amplified DNA allowing PCR analysis for codons 17 and 26 of the beta-globin gene that cause the beta-thalassemia disorder. We developed a rapid and highly specific technique of single-cell PCR to amplify a specific region on the beta-globin gene for codon 17 (AAG-->TAG) and codon 26 (GAG-->AAG) by using single-cell PCR. RESULTS: About 249 bp of amplicon for codon 17 and about 200 bp of amplicon for codon 26 were successfully amplified. No mutations were observed. Analyzed embryos were not transferred back to patients because the embryos used as samples were wasted embryos. CONCLUSIONS: Compared to other approaches for prenatal diagnosis, PGD is rapid and suitable as a noninvasive clinical tool for identifying genetic disorders for the purpose of reducing selective miscarriages and moral dilemmas. We opine that DNA extraction and amplification can be successfully performed by using single-cell PCR to diagnose genetic diseases before pregnancy.

Assessing the nutritional status of children with leukemia from hospitals in kuala lumpur.

A cross-sectional study was carried out to evaluate the nutritional status of 51 subjects with leukemia aged 4 to 12 years from the Haematology and Oncology Paediatric Ward, Universiti Kebangsaan Malaysia Medical Centre (PPUKM) and the Paediatric Institute of Kuala Lumpur. Nutritional status was assessed using anthropometric measurements, biochemical and haematological parameters. Subjects comprised 32 (62.7%) males and 19 (27.3%) females. Most of the subjects (41.2%) were in the age group of 4 to 6 years. More than half of the children were Malays (70.6%) followed by Indians (15.7%) and Chinese (13.7%). The subjects were diagnosed as acute lymphoblastic leukemia (ALL) (84.3%) followed by acute myelogenous leukemia (AML) (13.7%) and chronic myelogenous leukemia (CML) (2.0%) respectively. Most of the children were in remission status (54.9%). Underweight (<-2 SD for weight-for-age) was observed in 37.3% of the children while 17.6% of them were stunted (<-2 SD for height-for-age), and sign(s) of malnutrition (<-2 SD) for mid upper arm circumference (MUAC)-for-age was observed in 15.7% of the subjects. Approximately 20.0% of the subjects were in the severe malnutrition category with respect to low serum albumin levels (<3.5g/dl). All subjects had hemoglobin levels of less than the normal range. While the results indicated no significant differences in the nutritional status of subjects with leukemia at different stages of treatment, it was observed that the prevalence of malnutrition was higher in children with newly diagnosed leukemia. Thus, the nutritional status of children with leukemia should be monitored closely as there is a likelihood of deterioration owing to the disease.

Assessing the Nutritional Status of Children with Leukemia from Hospitals in Kuala Lumpur

A cross-sectional study was carried out to evaluate the nutritional status of 51 subjects with leukemia aged 4 to 12 years from the Haematology & Oncology Paediatric Ward, Universiti Kebangsaan Malaysia Medical Centre (PPUKM) and the Paediatric Institute of Kuala Lumpur. Nutritional status was assessed using anthropometric measurements, biochemical and haematological parameters. Subjects comprised 32 (62.7%) males and 19 (27.3%) females. Most of the subjects (41.2%) were in the age group of 4 to 6 years. More than half of the children were Malays (70.6%) followed by Indians (15.7%) and Chinese (13.7%). The subjects were diagnosed as acute lymphoblastic leukemia (ALL) (84.3%) followed by acute myelogenous leukemia (AML) (13.7%) and chronic myelogenous leukemia (CML) (2.0%) respectively. Most of the children were in remission status (54.9%). Underweight (<-2 SD for weight-for-age) was observed in 37.3% of the children while 17.6% of them were stunted (<-2 SD for height-for-age), and sign(s) of malnutrition (<-2 SD) for mid upper arm circumference (MUAC)-for-age was observed in 15.7% of the subjects. Approximately 20.0% of the subjects were in the severe malnutrition category with respect to low serum albumin levels (<3.5g/dl). All subjects had hemoglobin levels of less than the normal range. While the results indicated no significant differences in the nutritional status of subjects with leukemia at different stages of treatment, it was observed that the prevalence of malnutrition was higher in children with newly diagnosed leukemia. Thus, the nutritional status of children with leukemia should be monitored closely as there is a likelihood of deterioration owing to the disease.

Totally implantable vascular access devices for cystic fibrosis.

BACKGROUND: Totally implantable vascular access devices are widely used in people with cystic fibrosis to provide intermittent venous access for therapeutic infusions. Their use is associated with some complications such as thrombosis, embolism and infection. OBJECTIVES: To assess if totally implantable venous access devices are a safe and effective route for providing venous access for intermittent administration of intravenous antibiotics in people with cystic fibrosis, also to assess strategies to reduce possible complications of totally implantable venous access devices (e.g. anticoagulants to reduce the risk of thrombosis). SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group trials register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and abstract books of conference proceedings. Date of the most recent search: May 2003. SELECTION CRITERIA: Randomised and quasi-randomised controlled trials which compared the use of totally implantable venous access devices in people with cystic fibrosis to other means of vascular access, trials which compared the different types of these devices against each other and trials which assessed strategies to reduce complications of these devices. DATA COLLECTION AND ANALYSIS: No relevant trials were identified. MAIN RESULTS: No trials were included in this review. REVIEWER'S CONCLUSIONS: Totally implantable vascular access devices are widely used in people with cystic fibrosis to provide intermittent venous access for therapeutic infusions. Reports of their use in people with cystic fibrosis suggest that they are safe and effective. These reports also suggest that certain interventions might reduce the risk of complications; however, it is disappointing that these reports have not been assessed by randomised controlled trials. This systematic review identifies the need for a multicentre randomised controlled trial assessing both efficacy and possible adverse effects of totally implantable venous access devices in cystic fibrosis.