The Rapid Enumeration of a Mixed Culture of S. cerevisiae & L. plantarum in Beer Using Image-based Cytometry

Mixed cultures are prevalent in the food industry. From sour beer to sourdough bread, mix culture popularity is growing. A number of microbiological mixtures are used in these unique fermenting processes to create distinctive flavor profiles of consumers’ favorite foods. Although mixed cultures seem ubiquitous, they are often not well enumerated. The aim of this thesis was to create a novel rapid image-based cytometry method to enumerate mixed cultures in beer. Imaged-based cytometry can be used to rapidly enumerate mixed microbial cultures, as opposed to traditional plate counting methods that can take days to grow and count. A novel method was developed using the Nexcelom Cellometer X2 (X2) using fluorescent dyes and size exclusion automated counting on a mixed culture containing the microorganisms, lactic acid bacteria and yeast (L. plantarum and S. cerevisiae). Traditional spread plating using MRS and ADPA selective plates were used to validate this new methodology. The enumeration results were not found to be significantly different (P\u3e0.05) when analyzed by ANOVA comparing X2 to traditional plating for log(CFU/mL). The difference between the log(CFU/mL) of the X2 and traditional plating was also within a ±0.5 log(CFU/mL). The difference was noted as having a iv negligible impact on commercial brewing applications. The X2 counts were most precise when the log(CFU/mL) for yeast were between approximately 5 to 7 log(CFU/mL) and 6 to 8 log(CFU/mL) for lactic acid bacteria cells. These ranges are important as they encompass sour beer fermentation pitch rates and can be actively used by brewers today. Overall, the novel image-based cytometry method accomplished the goal of precisely and consistently counting a mixed culture (L. plantarum and S. cerevisiae) in beer media within the identified ranges and correlated well with the standard process of traditional plating methods. This technique enables brewers to make proactive decisions during fermentation, saves brewers time, and can be a cost effective alternative to other enumeration methods

Clinical Decision Support Systems (CDSS) applications in psychological suitability assessments.

Psychological suitability assessments are an integral component of public safety recruitment and selection processes. Psychological suitability assessments can benefit from the implementation of decision support systems. Providing an estimate of the likelihood for outcomes can act to support psychologists in decision making processes. This thesis critiques a content focused approach to psychological suitability assessment, develops a classification algorithm that estimates profile outcome likelihoods of several possible psychological suitability decision categories, and assesses the effect of providing estimates to psychologists on psychological decision making processes when determining profile outcomes. The critique outlines that the assessment model dimensions were developed to suit the needs of the organization at the time and connected psychological data with anticipated behaviours and activities. However, the model was not developed within a systematic process and relied heavily on subjective accounts of on duty officers. The assessment model could be improved with the implementation of new and updated tools, such as the MMPI-2RF or the MPULSE. The classification algorithm established by the discriminant function analysis accurately classified 86.75% of cases (p0.05). Experience conducting these types of assessments was a significant covariate for the time it took to render a decision, and the time it took to render a decision was not affected by the presentation of a likelihood estimate. Overall, this study suggests that decision support systems can be implemented to support a temporary clinical lead in navigating the psychological data such that candidates are evaluated more consistently for psychological suitability evaluations. Implementing a decision support system in practice can act as a guide for interpreting psychological data, reduce the error of both experienced and inexperienced assessors, and improve the integrity of the assessment.Master of Arts (MA) in Psycholog

Does Rubella Immunity Predict Measles Immunity? A Serosurvey of Pregnant Women

Background. This study was undertaken to determine whether rubella immunity infers measles immunity in pregnant women. Methods. Stored serum samples were obtained from the Iowa State Hygienic Laboratory for evaluation of rubella and measles immunities with IgG enzyme-linked immunosorbent assay. Results. Nine hundred serum samples were obtained for testing. The average age of the women at the time of antepartum serum collection was 28 (range, 14 to 44) years. Measles and rubella immunity were 88% and 98%, respectively; there was no effect of immunity status by age identified. Eighty eight percent of those with rubella immunity were also measles immune. There was no association between paired rubella and measles immunity identified, P < .0001. Discussion. Known rubella immunity did not infer measles immunity in our population. Thus, we recommend that pregnant women exposed to measles be tested and appropriately treated if they are found to be nonimmune

Effectiveness and safety of oral HIV preexposure prophylaxis for all populations.

ObjectivePreexposure prophylaxis (PrEP) offers a promising new approach to HIV prevention. This systematic review and meta-analysis evaluated the evidence for use of oral PrEP containing tenofovir disoproxil fumarate as an additional HIV prevention strategy in populations at substantial risk for HIV based on HIV acquisition, adverse events, drug resistance, sexual behavior, and reproductive health outcomes.DesignRigorous systematic review and meta-analysis.MethodsA comprehensive search strategy reviewed three electronic databases and conference abstracts through April 2015. Pooled effect estimates were calculated using random-effects meta-analysis.ResultsEighteen studies were included, comprising data from 39 articles and six conference abstracts. Across populations and PrEP regimens, PrEP significantly reduced the risk of HIV acquisition compared with placebo. Trials with PrEP use more than 70% demonstrated the highest PrEP effectiveness (risk ratio = 0.30, 95% confidence interval: 0.21-0.45, P &lt; 0.001) compared with placebo. Trials with low PrEP use did not show a significantly protective effect. Adverse events were similar between PrEP and placebo groups. More cases of drug-resistant HIV infection were found among PrEP users who initiated PrEP while acutely HIV-infected, but incidence of acquiring drug-resistant HIV during PrEP use was low. Studies consistently found no association between PrEP use and changes in sexual risk behavior. PrEP was not associated with increased pregnancy-related adverse events or hormonal contraception effectiveness.ConclusionPrEP is protective against HIV infection across populations, presents few significant safety risks, and there is no evidence of behavioral risk compensation. The effective and cost-effective use of PrEP will require development of best practices for fostering uptake and adherence among people at substantial HIV risk

Infrared colours and inferred masses of metal-poor giant stars in the Keplerfield

Intrinsically luminous giant stars in the Milky Way are the only potential volume-complete tracers of the distant disk, bulge, and halo. The chemical abundances of metal-poor giants also reflect the compositions of the earliest star-forming regions, providing the initial conditions for the chemical evolution of the Galaxy. However, the intrinsic rarity of metal-poor giants combined with the difficulty of efficiently identifying them with broad-band optical photometry has made it difficult to exploit them for studies of the Milky Way. One long-standing problem is that photometric selections for giant and/or metal-poor stars frequently include a large fraction of metal-rich dwarf contaminants. We re-derive a giant star photometric selection using existing public g-band and narrow-band DDO51photometry obtained in the Keplerfield. Our selection is simple and yields a contamination rate of main-sequence stars of ≲1% and a completeness of about 80 % for giant stars with Teff ≲ 5250 K - subject to the selection function of the spectroscopic surveys used to estimate these rates, and the magnitude range considered (11 ≲ g ≲ 15). While the DDO51filter is known to be sensitive to stellar surface gravity, we further show that the mid-infrared colours of DDO51-selected giants are strongly correlated with spectroscopic metallicity. This extends the infrared metal-poor selection developed by Schlaufman & Casey, demonstrating that the principal contaminants in their selection can be efficiently removed by the photometric separation of dwarfs and giants. This implies that any similarly efficient dwarf/giant discriminant (e.g., Gaiaparallaxes) can be used in conjunction with WISEcolours to select samples of giant stars with high completeness and low contamination. We employ our photometric selection to identify three metal-poor giant candidates in the Keplerfield with global asteroseismic parameters and find that masses inferred for these three stars using standard asteroseismic scaling relations are systematically over-estimated by 20-175%. Taken at face value, this small sample size implies that standard asteroseismic scaling relations over-predict stellar masses for metal-poor giant stars

A study of cultivation and sowing systems for cereals.

End of Project ReportDue to the pressure on cereal margins growers are seeking ways of reducing their costs of production. Reduced cultivation may provide cheaper and faster crop establishment, but in the past has been subject to problems with (poor crop establishment) weeds and soil compaction. With modern cultivation and sowing technology and methodology it may be possible to overcome these difficulties. In addition reduced cultivation is considered to be more environmentally friendly and sustainable than the conventional plough-based system. With these considerations in mind it was decided to start an experiment in autumn 2000 to compare the conventional system (PL) with reduced cultivation (RC) for crop establishment, and to examine the effect of incorporating the straw into then soil or baling and removing it. The treatments were assessed on winter wheat, and winter and spring barley. The parameters assessed included – crop establishment, yield and quality effects on disease levels, invertebrate pests and beneficials (aphids, slugs, earthworms, beetles), and soil conditions. The work reported here was conducted on four trial sites at Oak Park, Carlow and Knockbeg, Co. Laois from 2000 to 2003. the objectives were to establish the efficacy of RC compared with PL and to its effects on the parameters outlined above. Plant establishment was lower after RC in most of the experiments. Winter wheat yields were good irrespective of the number of plants established and there were no significant differences between cultivation or straw disposal method. The relative yields of the PL and RC treatments in winter barley varied over the three years; in 2001 there was no difference, in 2002 PL yielded 1 t/ha more than RC, while in 2003 the PL area produced 2 t/ha extra. Spring barley yields were similar on the PL and RC treatments. Broadleaved weeds were not a problem on the PL or RC treatments but there were more annual grass weeds on the RC plots. This was particularly serious in the winter barley experiments where sterile brome (Bromus sterilis) had a big effect on yield by 2003. annual meadow grass (Poa annua) was a problem in both PL and RC areas if not controlled by timely herbicide application; this was particularly so on the headland areas in the spring barley field. Disease assessment on the winter wheat trail showed no significant differences in Take-all or Eyespot levels between cultivation or straw disposal methods, although Take-all levels tended to be lower on the RC plots. Soil strength as measured by cone penetrometer and shear vane was higher on the RC treatments. In the winter wheat there was no difference in slug numbers between any of the treatments. Leaf damage by slugs increased in RC relative to PL but not significantly, in 2003. No below ground slug damage was found. Earthworm numbers in the winter barley increased significantly on the RC treatment relative to the PL as the study progressed. Straw incorporation increased earthworm numbers on both PL and RC treatments. Allolobophora chlorotia was the most common species in 2004. In the winter wheat the RC and straw incorporation increased earthworm numbers. Cultivation method had a greater effect on ground beetle numbers than method of straw disposal. Large species (e.g. Pterostichies malanarius) were favoured by RC while smaller species (e.g. Bembidon species) were more numerous in the PL plots. Molecular studies on virus diseases, vectors and vector-predators are underway in progress. In the invertebrate pests and beneficials investigations the RC winter barley had 11% fewer aphids and 27% less BYDV than the PL. Incorporating straw reduced aphid numbers and virus incidence by 36% and 32% respectively; the comparable values for the PL + straw treatment were 15% and 15%. In the winter wheat there was less BYDV in the RC plots than the PL + less were the straw was incorporated than where the straw was removed. Aphid infestation of wheat ears was low in the three years of the experiment. The insecticide seed treatment imidacloprid significantly reduced aphid numbers and virus incidence in winter barley but was less effective than a single insecticide spray in controlling the disease. The seed treatment was more effective in controlling BYDV in RC than in the PL plots. Estimations of slug numbers in the winter barley showed that the dominant species was the grey field slug (Deroceras reticulatum). Slug numbers increased significantly on all treatments between 2001 and 2004. In November 2004, slugs were significantly more numerous in the RC treatments than in the PL areas. Leaf damage was proportional to slug populations, but the slugs did not damage the barely seed or reduce plant populations in continuous winter barley under RC. Root and stem diseases (Take-all and Eyespot) were not nay worse under RC than after PL. In fact there was a tendency towards lower disease levels on the RC areas and there was significantly less sharp eyespot on the RC treatment

Reduction of leukocyte microvascular adherence and preservation of blood-brain barrier function by superoxide-lowering therapies in a piglet model of neonatal asphyxia

Background: Asphyxia is the most common cause of brain damage in newborns. Substantial evidence indicates that leukocyte recruitment in the cerebral vasculature during asphyxia contributes to this damage. We tested the hypothesis that superoxide radical (O2⋅_) promotes an acute post-asphyxial inflammatory response and blood-brain barrier (BBB) breakdown. We investigated the effects of removing O2⋅_ by superoxide dismutase (SOD) or C3, the cell-permeable SOD mimetic, in protecting against asphyxia-related leukocyte recruitment. We also tested the hypothesis that xanthine oxidase activity is one source of this radical.Methods: Anesthetized piglets were tracheostomized, ventilated, and equipped with closed cranial windows for the assessment of post-asphyxial rhodamine 6G-labeled leukocyte-endothelial adherence and microvascular permeability to sodium fluorescein in cortical venules. Asphyxia was induced by discontinuing ventilation. SOD and C3 were administered by cortical superfusion. The xanthine oxidase inhibitor oxypurinol was administered intravenously.Results: Leukocyte-venular adherence significantly increased during the initial 2 h of post-asphyxial reperfusion. BBB permeability was also elevated relative to non-asphyxial controls. Inhibition of O2⋅_ production by oxypurinol, or elimination of O2⋅_ by SOD or C3, significantly reduced rhodamine 6G-labeled leukocyte-endothelial adherence and improved BBB integrity, as measured by sodium fluorescein leak from cerebral microvessels.Conclusion: Using three different strategies to either prevent formation or enhance elimination of O2⋅_ during the post-asphyxial period, we saw both reduced leukocyte adherence and preserved BBB function with treatment. These findings suggest that agents which lower O2⋅_ in brain may be attractive new therapeutic interventions for the protection of the neonatal brain following asphyxia

Rocaglates as dual-targeting agents for experimental cerebral malaria

Cerebral malaria (CM) is a severe and rapidly progressing complication of infection by Plasmodium parasites that is associated with high rates of mortality and morbidity. Treatment options are currently few, and intervention with artemisinin (Art) has limited efficacy, a problem that is compounded by the emergence of resistance to Art in Plasmodium parasites. Rocaglates are a class of natural products derived from plants of the Aglaia genus that have been shown to interfere with eukaryotic initiation factor 4A (eIF4A), ultimately blocking initiation of protein synthesis. Here, we show that the rocaglate CR-1-31B perturbs association of Plasmodium falciparum eIF4A (PfeIF4A) with RNA. CR-1-31B shows potent prophylactic and therapeutic antiplasmodial activity in vivo in mouse models of infection with Plasmodium berghei (CM) and Plasmodium chabaudi (blood-stage malaria), and can also block replication of different clinical isolates of P. falciparum in human erythrocytes infected ex vivo, including drug-resistant P. falciparum isolates. In vivo, a single dosing of CR-1-31B in P. berghei-infected animals is sufficient to provide protection against lethality. CR-1-31B is shown to dampen expression of the early proinflammatory response in myeloid cells in vitro and dampens the inflammatory response in vivo in P. berghei-infected mice. The dual activity of CR-1-31B as an antiplasmodial and as an inhibitor of the inflammatory response in myeloid cells should prove extremely valuable for therapeutic intervention in human cases of CM.We thank Susan Gauthier, Genevieve Perreault, and Patrick Senechal for technical assistance. This work was supported by a research grant (to P.G.) from the Canadian Institutes of Health Research (CIHR) (Foundation Grant). J.P. and P.G. are supported by a James McGill Professorship salary award. D.L. is supported by fellowships from the Fonds de recherche sante Quebec, the CIHR Neuroinflammation training program. J.P. is supported by CIHR Research Grant FDN-148366. M.S. is supported by a CIHR Foundation grant. J.A.P. is supported by NIH Grant R35 GM118173. Work at the Boston University Center for Molecular Discovery is supported by Grant R24 GM111625. K.C.K. was supported by a CIHR Foundation Grant and the Canada Research Chair program. (Canadian Institutes of Health Research (CIHR); James McGill Professorship salary award; Fonds de recherche sante Quebec; CIHR Neuroinflammation training program; FDN-148366 - CIHR Research Grant; CIHR Foundation grant; R35 GM118173 - NIH; Canada Research Chair program; R24 GM111625