The Effect Of Moringa Oleifera On The Oncolytic Activity Of Vesicular Stomatitis Virus In Cervical Cancer Cells

Traditional treatment methods for cervical cancer are often not cancer-specific and are associated with adverse side effects. Our lab focuses on developing vesicular stomatitis virus (VSV) as an oncolytic agent due to the natural ability of this virus to target susceptible cancer cells. We are interested in augmenting the oncolytic activity of VSV by treating resistant cancer cells with natural compounds. We hypothesize that Moringa oleifera, which has previously demonstrated anticancer and anti-inflammatory properties, will promote killing of resistant cervical cancer cells by VSV by activating immune cells and promoting anti-tumor immunity. Of the five extracts of M. oleifera examined (aqueous, butanolic, ethanolic, hydroethanolic, and methanolic), the ethanolic extract promoted C4-II and HeLa cell killing alone and in combination with a wild-type strain (rwt) of VSV by decreasing activation of pro-survival pathways. The methanolic extract promoted killing of SiHa cervical cancer cells by the rM51R-M VSV strain. Furthermore, STAT1 levels decreased in VSV-infected cells, suggesting M. oleifera may inhibit induction of an antiviral response. M. oleifera stimulated maturation of dendritic cell (DC) subsets and pro-inflammatory cytokines following infection by VSV, indicating the pretreatment with M. oleifera has the potential to promote clearance of virus infected cervical cancer cells

Determinants of anemia and hemoglobin concentration in haitian school-aged children

Anemia diminishes oxygen transport in the body, resulting in potentially irreversible growth and developmental consequences for children. Limited evidence for determinants of anemia exists for school-aged children. We conducted a cluster randomized controlled trial in Haiti from 2012 to 2013 to test the efficacy of a fortified school snack. Children (N = 1,047) aged 3–13 years were followed longitudinally at three time points for hemoglobin (Hb) concentrations, anthropometry, and bioelectrical impedance measures. Dietary intakes, infectious disease morbidities, and socioeconomic and demographic factors were collected at baseline and endline. Longitudinal regression modeling with generalized least squares and logit models with random effects identified anemia risk factors beyond the intervention effect. At baseline, 70.6% of children were anemic and 2.6% were severely anemic. Stunting increased the odds of developing anemia (adjusted odds ratio [OR]: 1.48, 95% confidence interval [CI]: 1.05–2.08) and severe anemia (adjusted OR: 2.47, 95% CI: 1.30–4.71). Parent-reported vitamin A supplementation and deworming were positively associated with Hb concentrations, whereas fever and poultry ownership showed a negative relationship with Hb concentration and increased odds of severe anemia, respectively. Further research should explore the full spectrum of anemia etiologies in school children, including genetic causes

Constructing a recombinant model of the human pyruvate dehydrogenase complex

The human pyruvate dehydrogenase complex (PDC) is a large macromolecular assembly involved in the oxidative decarboxylation of pyruvate yielding acetyl CoA as the end product of this reaction, which subsequently enters the tricarboxylic acid (TCA) cycle. PDC is composed of multiple copies of various enzyme subunits, termed E1, E2 and E3. Human PDC also contains an additional component, E3BP, which has evolved in order to bind E3 to the core of the complex. The individual components of human PDC have now been cloned and overexpressed in E. coli. A His-tag has been engineered into the N-terminus of each protein to facilitate the rapid purification of these subunits using affinity chromatography. With the exception of E1, an alpha2beta2 heterotetramer, all recombinant proteins are soluble and produced in high yield. By using antibodies specific only for lipoylated E2 and E3BP from PDC, and by assaying the catalytically active subunits for activity, it has been found that these proteins are correctly folded and have been produced in active form. The use of a detergent, N-lauroylsarcosine, was required to produce a soluble E1. However, this component is active, as determined by enzymatic assay, under these conditions. Gel filtration studies have shown that E2 and E3BP must be coexpressed in E. coli in order for them to assemble into the stable E2/E3BP core complex that is central to the structure and organisation of human PDC. When these two proteins are expressed individually and then mixed they cannot form a stable core assembly. This suggests that the association between E2 and E3BP occurs in a co-translational manner and presumably requires their initial association as folding intermediates. Circular dichroism and fluorescence studies have been employed to examine the stability of the independently expressed E3BP. These studies have shown that each domain of E3BP, the N-terminal lipoyl domain, subunit-binding domain and C-terminal inner domain, unfolds at discrete concentrations of GdmCl. This indicates that while each domain is capable of independent folding, they also unfold independently of one another. In the native complex it is unclear how many E3 dimers are associated with human PDC. Isothermal titration calorimetry was utilised in order to assess the stoichiometry of binding between the recombinant E3BP and E3. These studies were performed using both full-length E3BP and a truncated construct, which consists of the subunit-binding domain expressed as a GST-fusion protein. These results suggest that one E3 dimer binds to two E3BP subunits; thus there would be six E3 dimers present per complex. The association constant for these two proteins was in the nanomolar range, indicative of very tight binding as expected. The binding affinity of E3 to E2 was also assessed using this technique. Truncated constructs of E2, specifically the subunit-binding domain expressed as a GST-fusion protein and the E2 didomain, a His-tagged protein containing the lipoyl domain and the subunit-binding domain of E2, were utilised in these studies. It was found that while E2 preferentially binds E1, it has also retained a residual affinity for the E3 subunit. Binding between E2 and E3 is approx. 100-1000 fold weaker than that between E3 and E3BP. The results described here support previous findings from our laboratory, obtained using an alternative technique, surface plasmon resonance and provide a molecular basis as to why E3BP- deficient patients retain residual PDC activity. While pursuing the main aim of this research, to reconstitute a recombinant human pyruvate dehydrogenase complex in vitro, a number of findings have produced interesting results. The unexpected 2:1 stoichiometry determined for the interaction between E3 and E3BP suggests that E3 dimers may form a network of crossbridges linking pairs of E3BP monomers across the 12 faces of the core. Production of sufficient quantities of active E1 is required in order to investigate whether a similar 2:1 stoichiometry exists between the alpha2beta2 E1 heterotetramer and the E2 didomain. If this is indeed the case, this introduces a new level of structure into the human pyruvate dehydrogenase complex, which has not been recognised previously

Young people's views about the purpose and composition of research ethics committees:findings from the PEARL qualitative study

BACKGROUND: Avon Longitudinal Study of Parents and Children (ALSPAC) is a birth cohort study within which the Project to Enhance ALSPAC through Record Linkage (PEARL) was established to enrich the ALSPAC resource through linkage between ALSPAC participants and routine sources of health and social data. PEARL incorporated qualitative research to seek the views of young people about data linkage, including their opinions about appropriate safeguards and research governance. In this paper we focus on views expressed about the purpose and composition of research ethics committees. METHODS: Digitally recorded interviews were conducted with 48 participants aged 17–19 years. Participants were asked about whether medical research should be monitored and controlled, their knowledge of research ethics committees, who should sit on these committees and what their role should be. Interview recordings were fully transcribed and anonymised. Thematic analysis was undertaken, assisted by the Framework approach to data management. RESULTS: The majority of interviewees had little or no specific knowledge of ethics committees. Once given basic information about research ethics committees, only three respondents suggested there was no need for such bodies to scrutinise research. The key tasks of ethics committees were identified as monitoring the research process and protecting research participants. The difficulty of balancing the potential to inhibit research against the need to protect research participants was acknowledged. The importance of relevant research and professional expertise was identified but it was also considered important to represent wider public opinion, and to counter the bias potentially associated with self-selection possibly through a selection process similar to ‘jury duty’. CONCLUSIONS: There is a need for more education and public awareness about the role and composition of research ethics committees. Despite an initial lack of knowledge, interviewees were able to contribute their ideas and balance the rights of individuals with the wider benefits from research. The suggestion that public opinion should be represented through random selection similar to jury duty may be worth pursuing in the light of the need to ensure diversity of opinion and establish trust amongst the general public about the use of ‘big data’ for the wider public good

Age-related mitochondrial DNA depletion and the impact on pancreatic beta cell function

Type 2 diabetes is characterised by an age-related decline in insulin secretion. We previously identified a 50% age-related decline in mitochondrial DNA (mtDNA) copy number in isolated human islets. The purpose of this study was to mimic this degree of mtDNA depletion in MIN6 cells to determine whether there is a direct impact on insulin secretion. Transcriptional silencing of mitochondrial transcription factor A, TFAM, decreased mtDNA levels by 40% in MIN6 cells. This level of mtDNA depletion significantly decreased mtDNA gene transcription and translation, resulting in reduced mitochondrial respiratory capacity and ATP production. Glucose-stimulated insulin secretion was impaired following partial mtDNA depletion, but was normalised following treatment with glibenclamide. This confirms that the deficit in the insulin secretory pathway precedes K+ channel closure, indicating that the impact of mtDNA depletion is at the level of mitochondrial respiration. In conclusion, partial mtDNA depletion to a degree comparable to that seen in aged human islets impaired mitochondrial function and directly decreased insulin secretion. Using our model of partial mtDNA depletion following targeted gene silencing of TFAM, we have managed to mimic the degree of mtDNA depletion observed in aged human islets, and have shown how this correlates with impaired insulin secretion. We therefore predict that the age-related mtDNA depletion in human islets is not simply a biomarker of the aging process, but will contribute to the age-related risk of type 2 diabetes

Randomized, Double-blinded, Placebo-controlled Trial of Amoxicillin/Clavulanic Acid to Prevent Preterm Delivery in Twin Gestation

Objective: The objective of this study was to determine whether prophylactic treatment with oral broad-spectrum antimicrobial therapy improves pregnancy outcomes in twin gestations

Evaluating Alternatives for Communicating About Food Risk

This article describes the development and preliminary evaluation of model materials designed as one-step in helping consumers understand how scientists assess food risk, how that information is used in food safety policy decisions and what individuals can do to protect themselves from residual risks

The Small Farmer-Tuskegee University-Walmart Project: Observations of the Steps within Commercial Supply

Abstract Observations of the various efforts necessary in an initiative, the Small Farmer-Tuskegee University-Walmart Project, to assist small farmers to comply with produce industry standards and supply produce to a major retailer over a six-year period were documented through an illustrative case study. The observations were taken from meetings with commercial buyers and farmers, site visits to processing centers and corporate farms, conference calls, and, mainly, from the authors’ “hands-on” participation with the functioning and preservation of this initiative. Consequently, these observations were organized into a framework of criteria that must be successively satisfied to be able to supply produce commercially. These criteria were capacity, capability, quality, food safety, consistency, sustainability, and marketability. A key finding was that for small farmers to meet these criteria, they required organization and support. It was concluded that although the effort was successful, the information gained through the effort was perhaps more valuable. Keywords: Small Farmers, Produce Markets, Commercial Supply, Capacit