Study of Bil-Uridine Diphosphate Glucuronyl Transferase (BILUDGPT- 1) Mutation in Neonatal Hyperbilirubinaemia with Glucose-6-Phosphate Dehydrogenase Deficiency.

Gilbert syndrome is caused by defects in the uridine diphosphate- glucuronosyltransferase 1A1 (UGT1A1) gene. These mutations differ among different populations and many of them have been found to be genetic risk factors for the development of neonatal jaundice

Preliminary comparative analysis of antibacterial effects of activated and non-activated of expired platelet concentrate by disc diffusion method

Background: Platelets release more than 30 cytokines to provide primary hemostatic function. In addition, platelets are also known to release antimicrobial peptides upon activation by thrombin. Materials and Methods: In this study, comparative analysis of antibacterial activity of activated and non-activated expired platelet concentrate was determined against Grampositive and Gram-negative bacteria by Kirby-Bauer disk diffusion method. Thrombin was used to prepare activated platelet concentrate. Gram-positive bacteria tested in this study were S.aureus and S.pyogenes and Gram-negative bacteria were E.coli and K.oxytoca. All the bacteria used in this study were sensitive strains from clinical isolates. Activated and nonactivated platelet showed no zone of inhibition against S.pyogenes and E.coli. Results: Activated platelet showed antibacterial activity against S.aureus and K.oxytoca with the zone of inhibition of 8.3 ± 0.6 mm and 7.7 ± 0.2 mm, respectively. Zone of inhibition observed in non-activated platelet against S.aureus and K.oxytoca were 7.8 ± 0.4 mm and 7.5 ± 0.3 mm, respectively. Conclusions: These findings showed that no significant differences in antibacterial activity produced by activated and non-activated platelet. However, zone of inhibition observed in activated and non-activated platelet indicate the presence of antibacterial property in expired platelet

Association of ABO blood groups with diabetes mellitus

Objective: So far no studies have been performed in Malaysia to look at association of diabetes mellitus (DM) with blood groups. We studied the association of ABO blood groups with DM type 2. Patients and methodology: It was a case control study conducted at Kepala Batas Hospital Batas, Penang, Malaysia in the year 2009, involving 70 patients with DM type 2 and 140 healthy controls. Ethical approval was obtained from Universiti Sains Malaysia. Blood samples were collected from the patients after consent. Samples were tested for ABO blood groups using ID-Card gel method. Results: Chi-square test results showed that there was an association between the ABO blood groups and DM type 2. It was found that A and O blood groups were negatively associated with DM type 2 (P<0.05) with higher percentage of A and O groups individuals were non-diabetic. No significant association was noted between DM type 2 and blood groups B (P=0.423) and AB (P=0.095). It was also noted that B blood group was distributed with highest percentage among patients with DM type 2 (53.71%) compared to controls (22.52%), but no statistical significance achieved. Conclusion: The results obtained suggest that there was a negative association between ABO blood groups A and O with DM type 2, with A and O group having less chances of diabetes. Large studies in other ethnic groups are needed to confirm these results

Sequence-specific inhibition of microRNA-130a gene by CRISPR/Cas9 system in breast cancer cell line

MicroRNAs (miRNAs) are short stranded noncoding RNA that play important roles in apoptosis, cell survival, development and cell proliferation. However, gene expression control via small regulatory RNA, particularly miRNA in breast cancer is still less explored. Therefore, this project aims to develop an approach to target microRNA-130a using the Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas9 system in MCF7, breast cancer cell line. The 20 bp sequences target at stem loop, 3' and 5' end of miR130a were cloned into pSpCas9(BB)-2AGFP (PX458) plasmid, and the positive clones were confirmed by sequencing. A total of 5 μg of PX458-miR130a was transfected to MCF7 using Lipofectamine® 3000 according to manufacturer’s protocol. The transfected cells were maintained in the incubator at 37 ⁰C under humidified 5% CO2. After 48 hours, cells were harvested and total RNA was extracted using miRNeasy Mini Kit (Qiagen). cDNAs were synthesised specific to miR-130a using TaqMan MicroRNA Reverse Transcription Kit (Applied Biosystems). Then, qRT-PCR was carried out using TaqMan Universal Master Mix (Applied Biosystems) to quantify the knockdown level of mature miRNAs in the cells. Result showed that miR-130a-5p was significantly downregulated in MCF7 cell line. However, no significant changes were observed for sequences targeting miR-130a-3p and stem loop. Thus, this study showed that the expression of miR-130a-5p was successfully down-regulated using CRISPR silencing system. This technique may be useful to manipulate the level of miRNA in various cell types to answer clinical questions at the molecular level

The study of glucose-6-phosphate dehydrogenase (G6PD) gene by restriction enzyme by restriction enzyme digestion in the Kelantan population

The most common diseases producing enzymopathy affecting the human population is glucose-6-phosphate dehydrogenase (G6PD) deficiency. It is estimated that about 400 million people worldwide are affected and this disease is commonest seen in the tropical and subtropical zones of the Eastern hemisphere. Molecular analysis has confirmed that the basis for G6PD deficiency is widely heterogeneous. Different mutants are responsible for the G6PD deficiency in the various parts of the world where this abnormality is prevalent. This study involved a sequential analysis whereby the blood from Malay neonates with neonatal jaundice admitted to Hospital Universiti Sains Malaysia and Kota Bharu Hospital were analyzed and polymerase chain reaction based analysis using serial multiplex primer method was done on those DNA samples. Samples that are found to be abnormal were then sequenced. Out of the 45 samples studied, 8 were found to have the Mediterranean mutation, two have the Mahidol mutation, two have Canton mutation and three have Kaiping mutation. Thus the molecular basis for the Malay neonatal jaundice in Kelantan is described with further prospect of population screening

De novo Ring Chromosome 6 in a Child with Multiple Congenital Anomalies

Ring chromosome 6, especially if it is de novo, is a rare occurrence. The phenotype of patients with ring chromosome 6 can be highly variable ranging from almost normal to severe malformations and mental retardation. The size and structure of the ring chromosome as well as the level of mosaicism are important factors in determining the clinical phenotype. Here we report an eight month-old child, a product of a non consanguineous marriage, who presented with developmental retardation, hypertelorism, microcephaly, flat occiput, broad nasal bridge, large ears, micrognathia, wide spaced nipples, protruding umbilicus, short stubby fingers, clinodactyly, single palmar crease, short neck with no obvious webbing, and congenital heart defect. Conventional karyotyping and Whole Chromosome Paint of the peripheral leukocytes showed 46,XY,r(6)(p25q27) karyotype with plausible breakpoints at p25 and q27 end. Conventional karyotyping of both parents showed normal karyotype. To the best of our knowledge, this is the first report of a Malay individual with ring chromosome 6, and this report adds to the collective knowledge of this rare chromosome abnormality

Dual-specificity phosphatase 6 (DUSP6): a review of its molecular characteristics and clinical relevance in cancer

Mitogen-activated protein kinases (MAPKs) are the main regulators of cellular proliferation, growth, and survival in physiologicalor pathological conditions. Aberrant MAPK signaling plays a pivotal role in carcinogenesis, which leads to development andprogression of human cancer. Dual-specificity phosphatase 6 (DUSP6), a member of the MAPK phosphatase family, interacts withspecifically targeted extracellular signal-regulated kinase 1/2 via negative feedback regulation in the MAPK pathway of mammaliancells. This phosphatase functions in a dual manner, pro-oncogenic or tumor-suppressive, depending on the type of cancer. Todate, the tumor-suppressive role of DUSP6 has been demonstrated in pancreatic cancer, non-small cell lung cancer, esophagealsquamous cell and nasopharyngeal carcinoma, and ovarian cancer. Its pro-oncogenic role has been observed in humanglioblastoma, thyroid carcinoma, breast cancer, and acute myeloid carcinoma. Both roles of DUSP6 have been documented inmalignant melanoma depending on the histological subtype of the cancer. Loss- or gain-of-function effects of DUSP6 in thesecancers highlights the significance of this phosphatase in carcinogenesis. Development of methods that use the DUSP6 gene as atherapeutic target for cancer treatment or as a prognostic factor for diagnosis and evaluation of cancer treatment outcome hasgreat potential. This review focuses on molecular characteristics of the DUSP6 gene and its role in cancers in the purview ofdevelopment, progression, and cancer treatment outcome

G6PD VIANGCHAN AND G6PD MEDITERRANEAN ARE THE MAIN VARIANTS I N G6PD DEFICIENCY IN THE MALAY POPULATION OF MALAYSIA

Abstract. Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked red blood cell enzymopathy common in malaria endemic areas. Individuals affected by this disease show a wide variety of clinical signs of acute hemolytic anemia. Mutations of the G6PD gene in the Malay population with G6PD deficiency in Kelantan, a state in North East Malaysia were studied. Ninety-three individuals with G6PD deficiency were subjected to mutation analysis of the G6PD gene using polymerase chain reaction based techniques of multiplex PCR. Of the ninety-three DNA samples studied, molecular defects were identified in 80 cases (86%). Variants were heterogeneous -28.7% were found to have a G to A nucleotide change at nucleotide 871 of the G6PD gene (G871A), corresponding to G6PD Viangchan. The other major mutations were G6PD Mediterranean, G6PD Vanua Lava, G6PD Coimbra, G6PD Kaiping, G6PD Orissa, G6PD Mahidol, G6PD Canton and G6PD Chatham. These results showed that there are heterogeneous mutations of the G6PD gene associated with G6PD deficiency and that G6PD Viangchan and G6PD Mediterranean account for the main variants in G6PD deficiency among the Malay population in Malaysia

Targeting Lung Cancer Stem Cells: Research and Clinical Impacts

Lung cancer is the most common cancer worldwide, accounting for 1.8 million new cases and 1.6 million deaths in 2012. Non-small cell lung cancer (NSCLC), which is one of two types of lung cancer, accounts for 85–90% of all lung cancers. Despite advances in therapy, lung cancer still remains a leading cause of death. Cancer relapse and dissemination after treatment indicates the existence of a niche of cancer cells that are not fully eradicated by current therapies. These chemoresistant populations of cancer cells are called cancer stem cells (CSCs) because they possess the self-renewal and differentiation capabilities similar to those of normal stem cells. Targeting the niche of CSCs in combination with chemotherapy might provide a promising strategy to eradicate these cells. Thus, understanding the characteristics of CSCs has become a focus of studies of NSCLC therapies

Conditioned Medium of Human Menstrual Blood-Derived Endometrial Stem Cells Protects Against MPP+-Induced Cytotoxicity in vitro

Mesenchymal stem cells (MSCs) showed the potential to treat Parkinson’s disease (PD). However, it is unknown whether the conditioned medium of human menstrual blood-derived endometrial stem cells (MenSCs-CM) has the function to alleviate syndromes of PD. In this study, human neuroblastoma SH-SY5Y cells were exposed to neurotoxicant 1-methyl-4-phenylpyridinium (MPP+) for inducing a range of response characteristics of PD. After culturing this cell model with 24 h/48 h collected MenSCs-CM for different days, cell viability, pro-inflammation cytokines, mitochondrial membrane potential (ΔΨm), oxidative stress, and cell apoptosis were detected. Finally, protein assay was performed to detect 12 kinds of neurotrophic factors inside MenSCs-CM. Our results showed that MPP+ caused SH-SY5Y cell viability reduction as an increasing dose and time dependent manner. MPP+ treatment resulted in inflammation, mitochondrial dysfunction, reactive oxygen species (ROS) production accumulation, and apoptosis of SH-SY5Y at its IC50 concentration. Forty-eight hours-collected MenSCs-CM and culturing with the MPP+-treated SH-SY5Y for 2 days are the optimized condition to increase cell viability. Besides, MenSCs-CM was efficacious against MPP+ induced inflammation, ΔΨm loss, ROS generation, and it could significantly decrease cells numbers in late apoptosis stage. What’s more, protein assay showed that MenSCs-CM contained various neuroprotective factors. Our study provided the first evidence that MenSCs-CM has a protective effect on MPP+-induced cytotoxicity in various aspects, and firstly showed that MenSCs can release at least 12 kinds of neurotrophic factors to medium, which may contribute to the protective function of MenSCs-CM to treat PD. This research enlightening that MenSCs-CM is beneficial in the therapy for PD and probably also for other neurodegenerative diseases