Field-Portable Immunoassay Instruments and Reagents to Measure Chelators and Mobile Forms of Uranium

Previous studies from our laboratory have demonstrated the feasibility of immunoassays for identification and quantification of specific metal ions. Our ultimate goal for this project is to (1) isolate and characterize antibodies that recognize the most mobile form of uranium, UO22+; (2) assemble, test, and validate a new field-portable immunosensor based on these antibodies; (3) prepare new monoclonal antibodies to the primary chelators (EDTA and DTPA) found in DOE wastes

Field-Portable Immunoassay Instruments and Reagents to Measure Chelators and Mobile Forms of Uranium

The goals for the 3-year project period are (1) to test and validate the present uranium sensor and develop protocols for its use at the NABIR Field Research Center; (2) to develop new reagents that will provide superior performance for the present hand-held immunosensor; and (3) to develop new antibodies that will permit this sensor to also measure other environmental contaminants (chromium, mercury, and/or DTPA). Sensor design modifications are underway via international collaborations. New reagents that will provide superior performance for the present hand-held immunosensor are being prepared and tested. New methods have been developed, to produce recombinant forms of metal-specific monoclonal antibodies for use with the sensor. Site-directed mutagenesis experiments are underway to determine the mechanisms of binding. Immunization experiments with sheep and rabbits to develop new recombinant forms of antibodies to metal-chelate complexes (chromium, mercury, and/or DTPA) have been initiated

Field-Portable Immunoassay Instruments and Reagents to Measure Chelators and Mobile Forms of Uranium

A collaborator in the Chemistry Department at Tulane University, Dr. Harry Ensley, has synthesized a new bifunctional chelator with specificity for ionic mercury (Hg2+) and methymercury (MeHg+). These chelators are based upon phenantholine derivative containing sulfhydryl functional groups. Experiments are underway to generate protein conjugates of this chelator to use in immunizations and screening

Resolution of nucleotide sugars and oligosaccharides by lectin affinity chromatography

Bandeiraea simplicifolia I (BS I) isolectins, immobilized on Sepharose beads, specifically retarded low molecular weight ligands containing terminal [alpha]--galactopyranosyl and 2-acetamido-2-deoxy-[alpha]--galactopyranosyl residues. A BS I lectin-Sepharose column has been used to perform very efficient separations of structurally homologous sugar nucleotides and oligosaccharides. For example, UDP-glucose and UDP-galactose (and the corresponding 2-acetamido-2-deoxy derivatives) were separated by as much as three column volumes by a BS I lectin-Sepharose column. This column has also been used to resolve the alditols of lactose ([beta]--Galp-(1 --> 4)--Glc) and melibiose ([alpha]--Galp-(1 --> 6)--Glc), to analyze the radiochemical purity of UDP-galactose and UDP-N-acetyl--galactosamine preparations, and to analyze the relative proportions of UDP-N-acetyl--glucosamine and UDP-N-acetyl--galactosamine in the UDP-N-acetyl--hexosamine pool isolated from cultured cells.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/23323/1/0000262.pd

Obesity Prevalence, Weight-Related Beliefs and Behaviors among Low-Income Ethnically Diverse National Job Corps Students

The obesity rates of Job Corps students, a predominantly ethnic minority and low income group of youth, are unknown. The purpose of this project was to examine obesity rates among these youth as well as their weight-related perceptions and behavior. First, medical charts (N=641) of all Job Corps students (ages 16-25) who were enrolled in the program in the past year were examined for height and weight. In the second phase of the study, 344 Job Corp students were recruited and information on weight perceptions, knowledge of obesity consequences, and weight loss behavior were examined. Almost half of the participants were overweight or obese. Overweight males were less likely to perceive themselves as being overweight than females. The majority of participants were aware of obesity-related health consequences but of those with past weight loss attempts, only 13% reported using both exercise and diet. High levels of overweight and obesity among Job Corps students are likely to impact employment and career goals. Evidence-based obesity interventions for Job Corps students are needed

Modification of cell membrane in variants of chinese hamster cells resistant to abrin

Stable and heritable variants of Chinese hamster ovary (CHO) cells which are resistant to different levels (0.1, 1.0 and 10 [mu]g/ml) of the toxin abrin have been isolated and characterized. The frequency of resistant colonies to abrin was increased with the concentration of a chemical mutagen. There was no effect of cell density or cross-feeding on the recovery of variants. In experiments using fluorescein-labeled abrin and ricin which bind to terminal (non-sialylated) galactose residues of cell-surface oligosaccharides, parental cells exhibited strong binding toward both toxins, whereas no fluorescence was observed in the resistant clones. A fluorescein-conjugated lectin, BS II, which is specific for terminal N-acetyl--glucosaminyl residues, did not interact with the parental cells, but did with the resistant clones. This suggests that on the surface of resistant cells the number of terminal galactosyl residues of oligosaccharide chains in glycoproteins was reduced, exposing the penultimate N-acetyl--glucosaminyl residues. The number of available endogenous acceptor sites for galactosyl transferase in the abrin-resistant clones was directly proportional to the degree of resistance. In the presence of great excess of exogenous acceptor, the rates of galactosyl transfer were similar in all the abrin-resistant cell types tested, with levels ranging from 1.4 to 1.7 times parental cell values. Studies with tetraploid cell hybrids reveal that resistance was a recessive trait. Fluctuation analysis showed that abrin resistance occurred in CHO cell populations at a rate of 4-7 x 10-8/cell/generation. The system may serve as a new marker for quantitative mutagenesis studies.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/23138/1/0000062.pd

Refined cut-off for TP53 immunohistochemistry improves prediction of TP53 mutation status in ovarian mucinous tumors: implications for outcome analyses.

TP53 mutations are implicated in the progression of mucinous borderline tumors (MBOT) to mucinous ovarian carcinomas (MOC). Optimized immunohistochemistry (IHC) for TP53 has been established as a proxy for the TP53 mutation status in other ovarian tumor types. We aimed to confirm the ability of TP53 IHC to predict TP53 mutation status in ovarian mucinous tumors and to evaluate the association of TP53 mutation status with survival among patients with MBOT and MOC. Tumor tissue from an initial cohort of 113 women with MBOT/MOC was stained with optimized IHC for TP53 using tissue microarrays (75.2%) or full sections (24.8%) and interpreted using established criteria as normal or abnormal (overexpression, complete absence, or cytoplasmic). Cases were considered concordant if abnormal IHC staining predicted deleterious TP53 mutations. Discordant tissue microarray cases were re-evaluated on full sections and interpretational criteria were refined. The initial cohort was expanded to a total of 165 MBOT and 424 MOC for the examination of the association of survival with TP53 mutation status, assessed either by TP53 IHC and/or sequencing. Initially, 82/113 (72.6%) cases were concordant using the established criteria. Refined criteria for overexpression to account for intratumoral heterogeneity and terminal differentiation improved concordance to 93.8% (106/113). In the expanded cohort, 19.4% (32/165) of MBOT showed evidence for TP53 mutation and this was associated with a higher risk of recurrence, disease-specific death, and all-cause mortality (overall survival: HR = 4.6, 95% CI 1.5-14.3, p = 0.0087). Within MOC, 61.1% (259/424) harbored a TP53 mutation, but this was not associated with survival (overall survival, p = 0.77). TP53 IHC is an accurate proxy for TP53 mutation status with refined interpretation criteria accounting for intratumoral heterogeneity and terminal differentiation in ovarian mucinous tumors. TP53 mutation status is an important biomarker to identify MBOT with a higher risk of mortality.KLG is supported by the Victorian Cancer Agency (MCRF15013) and the Australian National Health and Medical Research Council (APP1045783 and #628434). This study was supported by the Peter MacCallum Cancer Foundation. CS is supported by a University of Melbourne Postgraduate Scholarship. DDB is supported by National Health and Medical Research Council of Australia (NHMRC) grants APP1092856 and APP1117044 and by the US National Cancer Institute U54 programme (U54CA209978-04). ELG and SHK are supported through P50 CA136393-10. The following cohorts that contributed to the GAMuT study were supported as follows: CASCADE: Supported by the Peter MacCallum Cancer Foundation AOCS: The Australian Ovarian Cancer Study Group was supported by the U.S. Army Medical Research and Materiel Command under DAMD17-01-1-0729, The Cancer Council Victoria, Queensland Cancer Fund, The Cancer Council New South Wales, The Cancer Council South Australia, The Cancer Council Tasmania and The Cancer Foundation of Western Australia (Multi-State Applications 191, 211 and 182) and the National Health and Medical Research Council of Australia (NHMRC; ID400413 and ID400281). The Australian Ovarian Cancer Study gratefully acknowledges additional support from Ovarian Cancer Australia and the Peter MacCallum Foundation. The AOCS also acknowledges the cooperation of the participating institutions in Australia and acknowledges the contribution of the study nurses, research assistants and all clinical and scientific collaborators to the study. The complete AOCS Study Group can be found at www.aocstudy.org. We would like to thank all of the women who participated in these research programs. OVCARE receives core funding from The BC Cancer Foundation and the VGH and UBC Hospital Foundation. The Gynaecological Oncology Biobank at Westmead is a member of the Australasian Biospecimen Network-Oncology group, which was funded by the National Health and Medical Research Council Enabling Grants ID 310670 & ID 628903 and the Cancer Institute NSW Grants ID 12/RIG/1-17 & 15/RIG/1-16. COEUR: This study uses resources provided by the Canadian Ovarian Cancer Research Consortium’s - COEUR biobank funded by the Terry Fox Research Institute and managed and supervised by the Centre hospitalier de l’Université de Montréal (CRCHUM). The Consortium acknowledges contributions to its COEUR biobank from Institutions across Canada (for a full list see http://www.tfri.ca/en/research/translational-research/coeur/coeur_biobanks.aspx). The following cohorts that contributed to OTTA were supported as follows: AOV: Canadian Institutes of Health Research (MOP-86727), Cancer Research Society (19319). BAV: ELAN Funds of the University of Erlangen-Nuremberg; DOV: NCI/NIH R01CA168758. Huntsman Cancer Foundation and the National Cancer Institute of the National Institutes of Health under Award Number P30CA042014. HAW: U.S. National 19 Institutes of Health (R01-CA58598, N01-CN-55424 and N01-PC-67001); MAY: National Institutes of Health (R01-CA122443, P30-CA15083, P50-CA136393); Mayo Foundation; Minnesota Ovarian Cancer Alliance; Fred C. and Katherine B. Andersen Foundation; SEA: SEARCH team: Mitul Shah, Jennifer Alsopp, Mercedes Jiminez-Linan SEARCH funding: Cancer Research UK (C490/A16561), the Cancer Research UK Cambridge Cancer Centre and the National Institute for Health Research Cambridge Biomedical Research Centres. The University of Cambridge has received salary support for PDPP from the NHS in the East of England through the Clinical Academic Reserve. JBD: Cancer Research UK Institute Group Award UK A22905 and A15601; STA: NIH grants U01 CA71966 and U01 CA69417; SWE: Swedish Cancer foundation, WeCanCureCancer and årKampMotCancer foundation; TVA: Canadian Institutes of Health Research grant (MOP-86727) and NIH/NCI 1 R01CA160669- 01A1; VAN: M.S. Anglesio is funded through a Michael Smith Foundation for Health Research Scholar Award and the Janet D. Cottrelle Foundation Scholars program managed by the BC Cancer Foundation. The Vancouver study cohort (TVAN) is supported by BC’s Ovarian Cancer Research team (OVCARE), the BC Cancer Foundation and The VGH+UBC Hospital Foundation. WMH: National Health and Medical Research Council of Australia, Enabling Grants ID 310670 & ID 628903. Cancer Institute NSW Grants 12/RIG/1-17 & 15/RIG/1-16

Aripiprazole Augmentation in the Treatment of Military-Related PTSD with Major Depression: a retrospective chart review

<p>Abstract</p> <p>Background</p> <p>In this chart review, we attempted to evaluate the benefits of adding aripiprazole in veterans with military-related PTSD and comorbid depression, who had been minimally or partially responsive to their existing medications.</p> <p>Methods</p> <p>A retrospective chart review of patients who received an open-label, flexible-dose, 12- week course of adjunctive aripiprazole was conducted in 27 military veterans meeting DSM-IV criteria for PTSD and comorbid major depression. Concomitant psychiatric medications continued unchanged, except for other antipsychotics which were discontinued prior to initiating aripiprazole. The primary outcome variable was a change from baseline in the PTSD checklist-military version (PCL-M) and the Beck Depression Inventory (BDI-II).</p> <p>Results</p> <p>PTSD severity (Total PCL scores) decreased from 56.11 at baseline to 46.85 at 12-weeks (p < 0.0001 from Wilcoxon signed rank test) and the depression severity decreased from 30.44 at baseline to 20.67 at 12-weeks (p < 0.0001 from Wilcoxon signed rank test). Thirty seven percent (10/27) were considered responders, as defined by a decrease in total PCL scores of at least 20 percent and 19% (5/27) were considered as responders as defined by a decrease in total BDI score of at least 50%.</p> <p>Conclusions</p> <p>The addition of aripiprazole contributed to a reduction in both PTSD and depression symptomatology in a population that has traditionally demonstrated poor pharmacological response. Further investigations, including double-blind, placebo-controlled studies, are essential to confirm and further demonstrate the benefit of aripiprazole augmentation in the treatment of military related PTSD.</p