Assessment of in vitro sperm characteristics and their importance in the prediction of conception rate in a bovine timed-AI program

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Assessment of field fertility and several in vitro sperm characteristics following the use of different Angus sires in a timed-AI program with suckled Nelore cows

In order to provide information that may help researchers to understand the main cause(s) of differences in bull fertility frequently observed in field trials, this study aimed to investigate conception rates as well as several in vitro sperm characteristics of different sires of unknown fertility utilized in a Timed-AI (TAI) program. Suckled Nelore cows submitted to the same TAI protocol were allocated into eight breeding groups of approximately 120 animals each. Frozen semen doses from three Angus bulls and three different batches from each bull were utilized. Approximately 100 doses from each batch were used in TAI. Sires, batches and AI technicians were equally distributed across breeding groups. Cows were examined for pregnancy diagnosis 40 d after TAI. For in vitro sperm analyses, the same thawing procedure was repeated in the laboratory to mimic field conditions. The following in vitro sperm characteristics were assessed: computerized motility, thermal resistance, plasma and acrosomal membrane integrity, lipid peroxidation, morphology, morphometry and chromatin structure. No effect of breeding group, body condition score, AI technician and sire was observed. However, some significant differences among bulls were detected in laboratory analyses. Semen from sire presenting numerically lower (P > 0.05) pregnancy/AI also presented lower (P < 0.05) values in all sperm characteristics analyzed in thermal resistance test at 4 h (Total Motility, Progressive Motility, Average Path Velocity, Straight-Line Velocity, Curvilinear Velocity, Amplitude of Lateral Head Displacement, Beat Cross Frequency, Straightness, Linearity, and Percentage of Rapidly Moving Cells), higher (P < 0.05) Major and Total Defects in sperm morphological test, lower (P < 0.05) Length, Ellipticity and Fourier parameter (Fourier 0) in sperm morphometric analysis as well as higher (P < 0.05) chromatin heterogeneity. It was concluded that, although no bull effect was observed in the field experiment, the sire that presented numerically lower pregnancy/AI also presented lower semen quality according to the laboratory analyses performed. (C) 2012 Elsevier B.V. All rights reserved.

TNF receptors regulate vascular homeostasis in zebrafish through a caspase-8, caspase-2 and P53 apoptotic program that bypasses caspase-3

Although it is known that tumor necrosis factor receptor (TNFR) signaling plays a crucial role in vascular integrity and homeostasis, the contribution of each receptor to these processes and the signaling pathway involved are still largely unknown. Here, we show that targeted gene knockdown of TNFRSF1B in zebrafish embryos results in the induction of a caspase-8, caspase-2 and P53-dependent apoptotic program in endothelial cells that bypasses caspase-3. Furthermore, the simultaneous depletion of TNFRSF1A or the activation of NF-κB rescue endothelial cell apoptosis, indicating that a signaling balance between both TNFRs is required for endothelial cell integrity. In endothelial cells, TNFRSF1A signals apoptosis through caspase-8, whereas TNFRSF1B signals survival via NF-κB. Similarly, TNFα promotes the apoptosis of human endothelial cells through TNFRSF1A and triggers caspase-2 and P53 activation. We have identified an evolutionarily conserved apoptotic pathway involved in vascular homeostasis that provides new therapeutic targets for the control of inflammation- and tumor-driven angiogenesis.

A review of economic analysis in the vaccination program; decision making and governance: Adivisory Committee on Immunization Practices (ACIP)

The Advisory Committee on Immunization Practices (ACIP) develops written recommendations for the routine administration of vaccines to children and adults in the U.S. civilian population. The ACIP is the only entity in the federal government that makes such recommendations. ACIP elaborates on selection of its members and rules out concerns regarding its integrity, but fails to provide information about the importance of economic analysis in vaccine selection. ACIP recommendations can have large health and economic consequences. Emphasis on economic evaluation in health is a likely response to severe pressures of the federal and state health budget. This study describes the economic aspects considered by the ACIP while sanctioning a vaccine, and reviews the economic evaluations (our economic data) provided for vaccine deliberations. A five year study period from 2004 to 2009 is adopted. Publicly available data from ACIP web database is used. Drummond et al. (2005) checklist serves as a guide to assess the quality of economic evaluations presented. Drummond et al.'s checklist is a comprehensive hence it is unrealistic to expect every ACIP deliberation to meet all of their criteria. For practical purposes we have selected seven criteria that we judge to be significant criteria provided by Drummond et al. Twenty-four data points were obtained in a five year period. Our results show that out of the total twenty-four data point‘s (economic evaluations) only five data points received a score of six; that is six items on the list of seven were met. None of the data points received a perfect score of seven. Seven of the twenty-four data points received a score of five. A minimum of a two score was received by only one of the economic analyses. The type of economic evaluation along with the model criteria and ICER/QALY criteria met at 0.875 (87.5%). These three criteria were met at the highest rate among the seven criteria studied. Our study findings demonstrate that the perspective criteria met at 0.583 (58.3%) followed by source and sensitivity analysis criteria both tied at 0.541 (54.1%). The discount factor was met at 0.250 (25.0%).^ Economic analysis is not a novel concept to the ACIP. It has been practiced and presented at these meetings on a regular basis for more than five years. ACIP‘s stated goal is to utilize good quality epidemiologic, clinical and economic analyses to help policy makers choose among alternatives presented and thus achieve a better informed decision. As seen in our study the economic analyses over the years are inconsistent. The large variability coupled with lack of a standardized format may compromise the utility of the economic information for decision-making. While making recommendations, the ACIP takes into account all available information about a vaccine. Thus it is vital that standardized high quality economic information is provided at the ACIP meetings. Our study may provide a call for the ACIP to further investigate deficiencies within the system and thereby to improve economic evaluation data presented. ^

Criteria for implementation of optimum integration algorithm into the WRECKER program. Volume 1, summary final report.

National Highway Traffic Safety Administration, Washington, D.C.

Criteria for implementation of optimum integration algorithm into the WRECKER program. Technical final report. Volume II.

National Highway Traffic Safety Administration, Washington, D.C.

Safety compliance test program: FMVSS 212, 219, 301-75, NCAP, Accident Investigation Division data. Final report.

National Highway Traffic Safety Administration, Office of Research and Development, Washington, D.C.

Producibility guidelines : a blueprint for product integrity.

"August 1993."

Proceedings of the Second Workshop on Containment Integrity /

"SAND84-1514."

Ecological indicators for system-wide assessment of the greater everglades ecosystem restoration program

Developing scientifically credible tools for measuring the success of ecological restoration projects is a difficult and a non-trivial task. Yet, reliable measures of the general health and ecological integrity of ecosystems are critical for assessing the success of restoration programs. The South Florida Ecosystem Restoration Task Force (Task Force), which helps coordinate a multi-billion dollar multi-organizational effort between federal, state, local and tribal governments to restore the Florida Everglades, is using a small set of system-wide ecological indicators to assess the restoration efforts. A team of scientists and managers identified eleven ecological indicators from a field of several hundred through a selection process using 12 criteria to determine their applicability as part of a system-wide suite. The 12 criteria are: (1) is the indicator relevant to the ecosystem? (2) Does it respond to variability at a scale that makes it applicable to the entire system? (3) Is the indicator feasible to implement and is it measureable? (4) Is the indicator sensitive to system drivers and is it predictable? (5) Is the indicator interpretable in a common language? (6) Are there situations where an optimistic trend with regard to an indicator might suggest a pessimistic restoration trend? (7) Are there situations where a pessimistic trend with regard to an indicator may be unrelated to restoration activities? (8) Is the indicator scientifically defensible? (9) Can clear, measureable targets be established for the indicator to allow for assessments of success? (10) Does the indicator have specificity to be able to result in corrective action? (11) What level of ecosystem process or structure does the indicator address? (12) Does the indicator provide early warning signs of ecological change? In addition, a two page stoplight report card was developed to assist in communicating the complex science inherent in ecological indicators in a common language for resource managers, policy makers and the public. The report card employs a universally understood stoplight symbol that uses green to indicate that targets are being met, yellow to indicate that targets have not been met and corrective action may be needed and red to represent that targets are far from being met and corrective action is required. This paper presents the scientific process and the results of the development and selection of the criteria, the indicators and the stoplight report card format and content. The detailed process and results for the individual indicators are presented in companion papers in this special issue of Ecological Indicators.

Defining the Role of the Histone Methyltransferase, PR-Set7, in Maintaining the Genome Integrity of Drosophila Melanogaster

The complete and faithful duplication of the genome is essential to ensure normal cell division and organismal development. Eukaryotic DNA replication is initiated at multiple sites termed origins of replication that are activated at different time through S phase. The replication timing program is regulated by the S-phase checkpoint, which signals and repairs replicative stress. Eukaryotic DNA is packaged with histones into chromatin, thus DNA-templated processes including replication are modulated by the local chromatin environment such as post-translational modifications (PTMs) of histones.

One such epigenetic mark, methylation of lysine 20 on histone H4 (H4K20), has been linked to chromatin compaction, transcription, DNA repair and DNA replication. H4K20 can be mono-, di- and tri-methylated. Monomethylation of H4K20 (H4K20me1) is mediated by the cell cycle-regulated histone methyltransferase PR-Set7 and subsequent di-/tri- methylation is catalyzed by Suv4-20. Prior studies have shown that PR-Set7 depletion in mammalian cells results in defective S phase progression and the accumulation of DNA damage, which may be partially attributed to defects in origin selection and activation. Meanwhile, overexpression of mammalian PR-Set7 recruits components of pre-Replication Complex (pre-RC) onto chromatin and licenses replication origins for re-replication. However, these studies were limited to only a handful of mammalian origins, and it remains unclear how PR-Set7 impacts the replication program on a genomic scale. Finally, the methylation substrates of PR-Set7 include both histone (H4K20) and non-histone targets, therefore it is necessary to directly test the role of H4K20 methylation in PR-Set7 regulated phenotypes.

I employed genetic, cytological, and genomic approaches to better understand the role of H4K20 methylation in regulating DNA replication and genome stability in Drosophila melanogaster cells. Depletion of Drosophila PR-Set7 by RNAi in cultured Kc167 cells led to an ATR-dependent cell cycle arrest with near 4N DNA content and the accumulation of DNA damage, indicating a defect in completing S phase. The cells were arrested at the second S phase following PR-Set7 downregulation, suggesting that it was an epigenetic effect that coupled to the dilution of histone modification over multiple cell cycles. To directly test the role of H4K20 methylation in regulating genome integrity, I collaborated with the Duronio Lab and observed spontaneous DNA damage on the imaginal wing discs of third instar mutant larvae that had an alanine substitution on H4K20 (H4K20A) thus unable to be methylated, confirming that H4K20 is a bona fide target of PR-Set7 in maintaining genome integrity.

One possible source of DNA damage due to loss of PR-Set7 is reduced origin activity. I used BrdU-seq to profile the genome-wide origin activation pattern. However, I found that deregulation of H4K20 methylation states by manipulating the H4K20 methyltransferases PR-Set7 and Suv4-20 had no impact on origin activation throughout the genome. I then mapped the genomic distribution of DNA damage upon PR-Set7 depletion. Surprisingly, ChIP-seq of the DNA damage marker γ-H2A.v located the DNA damage to late replicating euchromatic regions of the Drosophila genome, and the strength of γ-H2A.v signal was uniformly distributed and spanned the entire late replication domain, implying stochastic replication fork collapse within late replicating regions. Together these data suggest that PR-Set7-mediated monomethylation of H4K20 is critical for maintaining the genomic integrity of late replicating domains, presumably via stabilization of late replicating forks.

In addition to investigating the function of H4K20me, I also used immunofluorescence to characterize the cell cycle regulated chromatin loading of Mcm2-7 complex, the DNA helicase that licenses replication origins, using H4K20me1 level as a proxy for cell cycle stages. In parallel with chromatin spindown data by Powell et al. (Powell et al. 2015), we showed a continuous loading of Mcm2-7 during G1 and a progressive removal from chromatin through S phase.

Academic Integrity and Senior Nursing Undergraduate Clinical Practice

Background: Academic integrity (AI) has been defined as the commitment to the values of honesty, trust, fairness, respect, and responsibility with courage in all academic endeavours. The senior years of nursing studies provide an intersection for students to transition to professional roles through student clinical practice. It is essential to understand what predicts senior nursing students’ intention to behave with AI so that efforts can be directed to initiatives focused on strengthening their commitment to behaving with AI. Research Questions: To what extent do students differ on Theory of Planned Behaviour (TPB) variables? What predicts intention to behave with academic integrity among senior nursing students in clinical practice across three different Canadian Schools of Nursing? Method: The TPB framework, an elicitation (n=30) and two pilot studies (n=59, n=29) resulted in the development of a 38 question (41-item) self-report survey (Miron Academic Integrity Nursing Survey—MAINS: α>0.70) that was administered to Year 3 and 4 students (N=339). Three predictor variables (attitude, subjective norm, perceived behavioural control) were measured with students’ intention to behave with AI in clinical. Age, sex, year of study, program stream, students’ understanding of AI policies, and locations where students accessed AI information were also measured. Results: Hierarchical multiple regression analyses revealed that background, site, and TPB variables explained 32.6% of the variance in intention to behave with academic integrity. The TPB variables explained 26.8% of the variance in intention after controlling for background and site variables. In the final model, only the TPB predictor variables were statistically significant with Attitude having the highest beta value (beta=0.35, p<0.001), followed by Subjective Norm (beta=0.21, p<0.001) and Perceived Behavioural Control (beta=0.12, p<0.02). Conclusion: Student attitude is the strongest predictor to intention to behave with AI in clinical practice and efforts to positively influence students’ attitudes need to be a focus for schools, curricula, and clinical educators. Opportunities for future research should include replicating the current study with students enrolled in other professional programs and intervention studies that examine the effectiveness of specific endeavours to promote AI in practice.

Chase-Lloyd House at 250: significance of function and integrity of form

(M. Historic Preservation) -- University of Maryland-College Park, 2015 Faculty Advisor: Dr. Constance Werner Ramirez. Program Director: Dr. Donald W. Linebaugh

Variable integrity testing of software

This paper provides a method of variable integrity testing to detect the faults of software. By checking the definition domain of the variable itself and the restriction condition of consistency between variables, we only need to determine the bound of each output data, rather than the exact value. Although the output of testing case cannot be easily caught, this method can improve the efficiency of detecting faults. Furthermore, the checking point is not limited to the final output place, but spreads over the reference places in the program code to key variables just like those at the checkpoints under debugging. As a consequence, the testing becomes more precise.