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Syfte: Syftet med studien är att studera i hur stor omfattning patienter är omgivna av andra i dödsögonblicket i palliativ vård och undersöka om det finns formulerade omvårdnadsmål i patienters IVP gällande vården i livets slutskede. Metod: Studien är kvantitativ retrospektiv med deskriptiv och analytisk design. I studien ingår samtliga cancerpatienter (n= 316), som vårdats och dött på en vårdavdelning och ASIH (avancerad sjukvård i hemmet) i Stockholm samt registrerats i Svenska palliativregistret (n=295). Resultat: Studien visade att i åldersgrupperna 75-84 år och 85 eller äldre var det fler som dog utan någon närvarande vid dödsögonblicket än i övriga åldersgrupper. Patienter dör utan att någon närvarande vid dödsögonblicket i ungefär lika stor omfattning i urvalet som på övriga palliativa enheter i Sverige. Resultatet visade att 62 % av de granskade journalerna hade omvårdnadsmål dokumenterade i IVP. Konklusion: Studien visar att närvaron vid dödsögonblicket sjunker när åldern stiger och att patienter dör utan någon närvarande vid dödsögonblicket i samma omfattning i urvalet som på övriga palliativa enheter i Sverige. Studien klarlägger även bristen på omvårdnadsmål i patienters IVP, vilket belyser ett förbättringsområde för att säkerställa kvaliteten på vården av patienter i livets slutskede. / Aims: The aim was to study the extent to which patients were surrounded by others at the time of death in palliative care and examine whether there were individual care plans, (IVP) in the patients nursing documentation regarding End-of-Life Care. Method: The study is quantitative retrospective with descriptive and analytical design. And includes all cancer patients (n = 316) who received care and died on a palliative care unit ward and ASIH (advanced medical care at home) in Stockholm and was registered in the Swedish palliative registry (n = 295). Results: In the age groups 75-84 years and 85 years or older, patients are more likely to die without someone present than in the other age groups. Patients are dying without anyone present in the same extent as in other palliative care units around Sweden. Only 62% of the sample had nursing goals documented in the patients IVP. Conclusion: The study illustrates that patients are surrounded by others in the same extent as in other palliative care units around Sweden. The study clarifies the lack of nursing goals in patients IVP, which highlights an area for improvement to ensure the quality in the End-of-Life care.

A predição do desempenho de touros na produção in vitro de embriões (PIVE), pela avaliação seminal, não está plenamente estabelecida. Com este estudo, objetivou-se caracterizar as diferenças nos atributos espermáticos de touros com baixo e alto desempenho, baseado nas taxas de clivagem e de blastocisto, visando estabelecer métodos preditivos de maior eficiência na PIVE. A partir do histórico de três anos da empresa In Vitro Brasil®, foram selecionados touros que haviam resultado nas menores e maiores taxas de clivagem e de blastocisto, estabelecendo-se 4 grupos experimentais: baixa taxa de clivagem (BC, n=5), alta taxa de clivagem (AC, n=5), baixa taxa de blastocisto (BB, n=5) e alta taxa de blastocisto (AB, n=5). Foram adquiridas palhetas de sêmen congelado dos touros de cada grupo, nas quais foram avaliados, antes (Pré-Percoll®) e após centrifugação em gradiente de Percoll® (Pós-Percoll®), os atributos espermáticos: cinética, em sistema CASA; integridade de membranas plasmática e acrossomal (FITC-PSA/PI), potencial de membrana mitocondrial (JC-1), status oxidativo (DHE, CellROXTM green/PI, MitoSox red), apoptose (FITC-FMK-VAD/PI) e DNA (SCSA), por citometria de fluxo; morfologia; atividade mitocondrial (DAB); capacitação (CTC); teste funcional e TBARS induzido. Diferenças entre grupos (BC vs. AC e BB vs. AB) foram estimadas pelo teste t. Para todas as análises estatísticas foi considerado nível de significância de 5%. Na ausência de interações significativas, o efeito dos grupos foi analisado ao fundir os momentos Pré e Pós-Percoll®. Os grupos AC e AB apresentaram valores superiores de VAP, VCL, ALH e DNC e valores inferiores de BCF, STR, LIN e WOB, cinética indicativa de hiperativação. Adicionalmente, estes grupos apresentaram indícios de menor ocorrência de estresse oxidativo, evidenciados pelo percentual de espermatozoides com membrana lesada e estresse oxidativo (CellROXTM green/PI) no grupo AC e nível de DHE no grupo AB. Para os demais atributos estudados, não se obteve diferença entre os grupos. Por meio de análise de regressão logística multivariada, constatou-se que a taxa de clivagem sofre efeito de: VCL, BCF e percentual de espermatozoides com integridade de membrana e sem estresse oxidativo; já a taxa de blastocisto, sofre efeito de: defeitos menores, VSL, DABIII e DHE, os quais foram inclusos nos respectivos modelos preditivos. As taxas obtidas com o modelo preditivo foram comparadas às taxas reais por análise de correlação de Pearson, que resultaram em intensidade moderada tanto para clivagem (r=0,56) quanto para blastocisto (r=0,44). Subpopulações espermáticas com diferentes perfis de cinética em CASA, foram determinadas em três etapas estatísticas segundo Núñez-Martínez et al. (2006). Foram detectadas 4 subpopulações: 1-Rápidos e progressivos; 2- Hiperativados; 3- Lentos e sinuosos; 4- Lentos e progressivos. Touros com alto desempenho (AC e AB) apresentaram maior percentual de subpopulação 2, Pré-Percoll®, e das subpopulações 1 e 2, Pós-Percoll®. Com isto, demonstra-se ser possível diferenciar touros de baixo e alto desempenho na PIVE, pela avaliação dos atributos espermáticos, com o estabelecimento de modelos preditivos para as taxas de clivagem e de blastocisto. Ademais, o estudo das subpopulações espermáticas constitui-se no método mais efetivo para a predição do desempenho in vitro. / The prediction of bull performance on the in vitro production (IVP) of bovine embryos, by the seminal evaluation, is not fully established. Thus, the aim of this study was to characterize the differences in the sperm features of bulls with low and high performance, based on cleavage and blastocyst rates, in order to establish predictive methods for a greater efficiency in the IVP. Based on the results obtained in three years from the company In Vitro Brasil®, bulls were selected by the lowest and highest cleavage and blastocyst rates, establishing 4 experimental groups: low cleavage rate (BC, n=5), high cleavage rate (AC, n=5), low blastocyst rate (BB, n=5) and high blastocyst rate (AB, n=5). Frozen semen were obtained from the bulls of each group, in which were evaluated, before (Pre-Percoll®) and after Percoll® gradient centrifugation (Post-Percoll®), the sperm features: kinetics, in CASA; integrity of plasma and acrosomal membranes (FITC-PSA/PI), mitochondrial membrane potential (JC-1), oxidative status (DHE, CellROXTM green/PI, MitoSox red), apoptosis (FITC-FMK-VAD/PI) and DNA (SCSA), by flow cytometry; morphology; mitochondrial activity (DAB); capacitation (CTC); functional test and induced TBARS. Differences between groups (BC vs. AC and BB vs. AB) were estimated by t test. A significance level of 5% was considered for all statistical analysis. In the absence of significant interactions, the effect of the groups was analyzed by merging the moments Pre and Post-Percoll®. The AC and AB groups presented higher values of VAP, VCL, ALH and DNC and lower values of BCF, STR, LIN and WOB, kinetics indicative of hyperactivation. In addition, these groups showed less evidence of oxidative stress, observed by the percentage of spermatozoa with damaged membrane and oxidative stress (CellROXTM green/PI) in the AC group and level of DHE in the AB group. For the others sperm features, there was no difference between groups. By multivariate logistic regression analysis, it was verified that the cleavage rate is affected by: VSL, BCF and percentage of spermatozoa with membrane integrity and without oxidative stress; and the blastocyst rate, is affected by: minor defects, VSL, DABIII and DHE, which were included in the respective predictive models. The rates obtained with the predictive model were compared to the real rates by Pearson correlation analysis, which resulted in moderate intensity for both cleavage (r=0.56) and blastocyst (r=0.44). Spermatic subpopulations with different kinetic profiles at CASA were determined in three statistical steps according to Núñez-Martínez et al. (2006). Four subpopulations were detected: 1-Fast and progressive; 2- Hyperactivated; 3- Slow and sinuous; 4- Slow and progressive. Bulls with high performance (AC and AB) presented higher percentage of subpopulation 2, Pre-Percoll®, and subpopulations 1 and 2, Post-Percoll®. In summary, it is possible to differentiate low and high-performance bulls at IVP by evaluating the sperm features, with the establishment of predictive models for the cleavage and blastocyst rates. Furthermore, the study of sperm subpopulations is the most effective method for predicting in vitro performance.

A predição do desempenho de touros na produção in vitro de embriões (PIVE), pela avaliação seminal, não está plenamente estabelecida. Com este estudo, objetivou-se caracterizar as diferenças nos atributos espermáticos de touros com baixo e alto desempenho, baseado nas taxas de clivagem e de blastocisto, visando estabelecer métodos preditivos de maior eficiência na PIVE. A partir do histórico de três anos da empresa In Vitro Brasil®, foram selecionados touros que haviam resultado nas menores e maiores taxas de clivagem e de blastocisto, estabelecendo-se 4 grupos experimentais: baixa taxa de clivagem (BC, n=5), alta taxa de clivagem (AC, n=5), baixa taxa de blastocisto (BB, n=5) e alta taxa de blastocisto (AB, n=5). Foram adquiridas palhetas de sêmen congelado dos touros de cada grupo, nas quais foram avaliados, antes (Pré-Percoll®) e após centrifugação em gradiente de Percoll® (Pós-Percoll®), os atributos espermáticos: cinética, em sistema CASA; integridade de membranas plasmática e acrossomal (FITC-PSA/PI), potencial de membrana mitocondrial (JC-1), status oxidativo (DHE, CellROXTM green/PI, MitoSox red), apoptose (FITC-FMK-VAD/PI) e DNA (SCSA), por citometria de fluxo; morfologia; atividade mitocondrial (DAB); capacitação (CTC); teste funcional e TBARS induzido. Diferenças entre grupos (BC vs. AC e BB vs. AB) foram estimadas pelo teste t. Para todas as análises estatísticas foi considerado nível de significância de 5%. Na ausência de interações significativas, o efeito dos grupos foi analisado ao fundir os momentos Pré e Pós-Percoll®. Os grupos AC e AB apresentaram valores superiores de VAP, VCL, ALH e DNC e valores inferiores de BCF, STR, LIN e WOB, cinética indicativa de hiperativação. Adicionalmente, estes grupos apresentaram indícios de menor ocorrência de estresse oxidativo, evidenciados pelo percentual de espermatozoides com membrana lesada e estresse oxidativo (CellROXTM green/PI) no grupo AC e nível de DHE no grupo AB. Para os demais atributos estudados, não se obteve diferença entre os grupos. Por meio de análise de regressão logística multivariada, constatou-se que a taxa de clivagem sofre efeito de: VCL, BCF e percentual de espermatozoides com integridade de membrana e sem estresse oxidativo; já a taxa de blastocisto, sofre efeito de: defeitos menores, VSL, DABIII e DHE, os quais foram inclusos nos respectivos modelos preditivos. As taxas obtidas com o modelo preditivo foram comparadas às taxas reais por análise de correlação de Pearson, que resultaram em intensidade moderada tanto para clivagem (r=0,56) quanto para blastocisto (r=0,44). Subpopulações espermáticas com diferentes perfis de cinética em CASA, foram determinadas em três etapas estatísticas segundo Núñez-Martínez et al. (2006). Foram detectadas 4 subpopulações: 1-Rápidos e progressivos; 2- Hiperativados; 3- Lentos e sinuosos; 4- Lentos e progressivos. Touros com alto desempenho (AC e AB) apresentaram maior percentual de subpopulação 2, Pré-Percoll®, e das subpopulações 1 e 2, Pós-Percoll®. Com isto, demonstra-se ser possível diferenciar touros de baixo e alto desempenho na PIVE, pela avaliação dos atributos espermáticos, com o estabelecimento de modelos preditivos para as taxas de clivagem e de blastocisto. Ademais, o estudo das subpopulações espermáticas constitui-se no método mais efetivo para a predição do desempenho in vitro. / The prediction of bull performance on the in vitro production (IVP) of bovine embryos, by the seminal evaluation, is not fully established. Thus, the aim of this study was to characterize the differences in the sperm features of bulls with low and high performance, based on cleavage and blastocyst rates, in order to establish predictive methods for a greater efficiency in the IVP. Based on the results obtained in three years from the company In Vitro Brasil®, bulls were selected by the lowest and highest cleavage and blastocyst rates, establishing 4 experimental groups: low cleavage rate (BC, n=5), high cleavage rate (AC, n=5), low blastocyst rate (BB, n=5) and high blastocyst rate (AB, n=5). Frozen semen were obtained from the bulls of each group, in which were evaluated, before (Pre-Percoll®) and after Percoll® gradient centrifugation (Post-Percoll®), the sperm features: kinetics, in CASA; integrity of plasma and acrosomal membranes (FITC-PSA/PI), mitochondrial membrane potential (JC-1), oxidative status (DHE, CellROXTM green/PI, MitoSox red), apoptosis (FITC-FMK-VAD/PI) and DNA (SCSA), by flow cytometry; morphology; mitochondrial activity (DAB); capacitation (CTC); functional test and induced TBARS. Differences between groups (BC vs. AC and BB vs. AB) were estimated by t test. A significance level of 5% was considered for all statistical analysis. In the absence of significant interactions, the effect of the groups was analyzed by merging the moments Pre and Post-Percoll®. The AC and AB groups presented higher values of VAP, VCL, ALH and DNC and lower values of BCF, STR, LIN and WOB, kinetics indicative of hyperactivation. In addition, these groups showed less evidence of oxidative stress, observed by the percentage of spermatozoa with damaged membrane and oxidative stress (CellROXTM green/PI) in the AC group and level of DHE in the AB group. For the others sperm features, there was no difference between groups. By multivariate logistic regression analysis, it was verified that the cleavage rate is affected by: VSL, BCF and percentage of spermatozoa with membrane integrity and without oxidative stress; and the blastocyst rate, is affected by: minor defects, VSL, DABIII and DHE, which were included in the respective predictive models. The rates obtained with the predictive model were compared to the real rates by Pearson correlation analysis, which resulted in moderate intensity for both cleavage (r=0.56) and blastocyst (r=0.44). Spermatic subpopulations with different kinetic profiles at CASA were determined in three statistical steps according to Núñez-Martínez et al. (2006). Four subpopulations were detected: 1-Fast and progressive; 2- Hyperactivated; 3- Slow and sinuous; 4- Slow and progressive. Bulls with high performance (AC and AB) presented higher percentage of subpopulation 2, Pre-Percoll®, and subpopulations 1 and 2, Post-Percoll®. In summary, it is possible to differentiate low and high-performance bulls at IVP by evaluating the sperm features, with the establishment of predictive models for the cleavage and blastocyst rates. Furthermore, the study of sperm subpopulations is the most effective method for predicting in vitro performance.

Submitted by Jordan (jordanbiblio@gmail.com) on 2017-03-02T14:03:04Z No. of bitstreams: 1 DISS_2015_Larissa Alves Berté Barbosa.pdf: 410769 bytes, checksum: 903677a2ce06bd213b48a06034419b79 (MD5) / Approved for entry into archive by Jordan (jordanbiblio@gmail.com) on 2017-03-03T12:03:08Z (GMT) No. of bitstreams: 1 DISS_2015_Larissa Alves Berté Barbosa.pdf: 410769 bytes, checksum: 903677a2ce06bd213b48a06034419b79 (MD5) / Made available in DSpace on 2017-03-03T12:03:08Z (GMT). No. of bitstreams: 1 DISS_2015_Larissa Alves Berté Barbosa.pdf: 410769 bytes, checksum: 903677a2ce06bd213b48a06034419b79 (MD5) Previous issue date: 2015-03-19 / O hormônio de crescimento (GH) é um hormônio secretado pela hipófise e atua no crescimento de vários tecidos, incluindo a sistema reprodutor. Ele não é um hormônio reprodutivo, porém tem uma ligação importante para o desenvolvimento dos folículos ovarianos e no amadurecimento oocitário. Seus receptores estão presentes nas células do cumulus na membrana citoplasmática e nuclear do oócito, e isso faz com que o GH atue diretamente em seu crescimento. Na produção in vitro de embriões o GH vem sendo estudado visando aumentar a quantidade e a qualidade dos embriões, porém a concentração de GH na maturação in vitro ainda não está definida, havendo uma variação na literatura de 10 a 1000 ng/mL. Com isso, esse estudo comparou diferentes dosagens de GH adicionados ao meio de maturação, a fim de definir uma dosagem ideal para que haja um aumento na quantidade e qualidade dos embriões produzidos, que foi mensurada através das taxas de clivagem, produção de embriões e quantificação do número de células embrionárias, como também se comparou a interação do GH com a geração de estresse oxidativo. Os oócitos foram maturados com meio composto por TCM 199 com sais de Earl, suplementado com 10% de soro fetal bovino, hormônio luteinizante, hormônio folículo estimulante, estradiol e amicacina. Diferentes dosagens de GH foram adicionadas ao meio de maturação sendo elas 0 ng/mL, 25 ng/mL, 50 ng/mL, 75 ng/mL e 100 ng/mL. Após 24 horas de maturação os oócitos foram fertilizados e estes foram incubados por 22 horas, para posteriormente passarem para o cultivo, onde ficaram incubadas por 7 dias. No terceiro dia de cultivo foram avaliadas as clivagens e no sétimo dia a produção de embriões. Os embriões em estagio de blastocisto expandido foram fixados em lâmina e corados com Panótico para a contagem de células embrionárias. A análise de estresse oxidativo foi feito pela reação dos meios de maturação, fertilização e cultivo com o acido tiobarbitúrico. Houve diferenças significativas sobre a quantificação de células quando usado 100 ng/ml proporcionando uma melhora na qualidade dos embriões, e também se aumentou o estresse oxidativo quando usado 75 e 100 ng/mL na fertilização e no cultivo in vitro. / Growth hormone (GH) is a hormone secreted by the pituitary gland and acts on the growth of various tissues, including the reproductive system. It is not a reproductive hormone, but has an important link to the development of ovarian follicles and the oocyte maturation. Its receptors are present in cumulus cells in cytoplasmic and nuclear membrane of the oocyte, and this causes the GH acts directly in its growth. In vitro production of embryos GH has been studied to increase the amount and quality of embryos, but the concentration of GH in vitro maturation is not yet defined, with a variation in the literature 10 to 1000 ng/mL. This study compared different dosages of GH added to the maturation medium in order to set an optimal dosage so that there is an increase in the quantity and quality of embryos produced, which was measured by the cleavage rates, embryo production and quantifying the number of embryonic stem cells, as well as comparing the interaction of GH with the generation of oxidative stress. Oocytes were matured in medium composed of TCM 199 with Earl's salts, supplemented with 10% fetal bovine serum, luteinizing hormone, follicle stimulating hormone, estradiol, and amikacin. Different dosages of GH were added to maturation medium these being 0 ng/mL 25 ng /mL, 50 ng/mL, 75 ng/mL and 100 ng/mL. After 24 hours of maturation, oocytes were fertilized and these were incubated for 22 hours to later pass to the culture and they were incubated for 7 days. On the third day of cultivation were evaluated cleavages and on the seventh day the production of embryos. Embryos expanded blastocyst stage were fixed on slides and stained with Panotic for embryonic cell count. The oxidative stress analysis was done by the reaction of the means of maturation, fertilization and cultivation with thiobarbituric acid. There were significant differences in quantitation of cells when used 100 ng/mL giving an improvement in the quality of embryos, and also increased the oxidative stress used when 75 and 100 ng/mL in fertilization and in vitro culture.

Cryopreservation of in vivo derived Jersey bovine embryos have resulted in a 10% lower pregnancy rate compared to other dairy breeds. Poor embryo survival after cryopreservation has been partially attributed to the high lipid content of Jersey embryos. In vitro-produced (IVP) bovine embryos have darker cytoplasm than their in vivo-derived counterparts because of higher lipid accumulation. High lipid accumulation is associated with impaired embryo quality. Forskolin is an adenylate cyclase activator that regulates cAMP levels in cells and has been shown to induce lipolysis in IVP embryos. L-carnitine is required for transport of fatty acids from the intermembrane space of the mitochondria into the mitochondrial matrix to support the process of β-oxidation, and enhances ATP production. We hypothesized that the lipid content of in vivo-produced and IVP Jersey embryos is higher than respective Holstein embryos and that forskolin + L-carnitine would reduce lipid content of IVP embryos and vitrification with embryo collapse would improve the cryosurvival of Jersey IVP embryos. The objectives of this experiment were (1) to analyze lipid content of in vivo and IVP Jersey and Holstein cattle embryos, (2) to evaluate the effect of forskolin + L-carnitine added to IVP culture media, and (3) evaluate Jersey IVP survival rates after three cryopreservation procedures. The factorial experimental design for objectives one and two used two breeds (Holstein and Jersey) and three embryo production methods (in vivo, IVP, and IVP + forsk/L-C). In vivo produced embryos (n = 27 blastocysts) were collected from superstimulated donors by routine procedures 7.5 days after AI. IVP embryos (n = 259 blastocysts) were produced by standard procedures; briefly, oocytes were aspirated from 2- to 8-mm follicles from slaughterhouse ovaries and matured for 24 h in SMM medium (BoviPro, MOFA Global, Verona, WI, USA). Matured oocytes were fertilized using semen from two different bulls for each breed, and embryos were cultured in BBH7 medium (BoviPro, MOFA Global) alone or with the addition of 1.5mM L-carnitine during maturation and embryo culture with forskolin (10 µM) added at Day 5 of culture at 38.5°C in 5% O2, 5% CO2, and 90% N2. The lipid content of embryos was quantified by staining Day 7 blastocysts with 1 μg mL–1 Nile red dye (580–596 nm), after which a digital photograph of the equatorial part of the embryo was taken at 40×, and fluorescence intensity (FI) was measured with Image Pro software. Data was analyzed by ANOVA, and means were compared using Tukey’s HSD. For the third objective, grade 1 Jersey IVP blastocysts (n=356) were divided into six treatments using a 2x3 factorial design comparing intact (IB) vs collapsed blastocoele (CB) and three cryopreservation methods: slow freezing (SF) vs vitrification using open pulled straws (OPS) or cryotop (CT). Slow freezing embryos were equilibrated in 0.7 M glycerol and 0.1 M galactose in holding media for 10 min, held for 10 min at -6°C, seeded after 5 min, decreased to -32 °C at 0.5 °C /min, held at -32°C for 5 min, and finally plunged into liquid nitrogen. Vitrified embryos were equilibrated in 1.5 M ethylene glycol (EG) for 5 min, exposed to 7 M EG + 0.6 M galactose for 30 s while loaded into OPS or placed onto CT, then immediately plunged into liquid nitrogen. SF embryos were thawed in air for 10 s and placed in a water bath at 37°C for 45 s. Vitrified embryos were warmed directly into holding medium at 37°C supplemented with 1.0 M, 0.5 M and 0.25 M galactose for 3 minutes each. Subsequently, embryos were cultured in BBH7 and re-expansion rates were assessed at 24 and 48h post warming and data was evaluated by GLIMX. For objective 1, Jersey and Holstein IVP embryos had higher lipid content than Holstein in vivo-produced embryos (P < 0.05), but were not different than Jersey in vivo-derived embryos (P > 0.1). Forskolin + L-carnitine lowered the lipid content (P < 0.05) of both IVP Jersey and Holstein embryos and was not different (P > 0.1) than in vivo-produced embryos. For experiment 2, re-expansion rates were higher for CT, than OPS, and SF (85 vs. 66 vs. 72% ± 0.4, respectively; p<0.05). Main effect means for re-expansion were higher for CB than IB (79 vs 68% ± 0.3; p<0.05). In conclusion, IVP embryos have higher lipid accumulation over Holstein in vivo embryos. Addition of forskolin and L-carnitine to embryo culture media has the potential to lower embryo lipid accumulation and possibly improve embryo viability and cryotolerance of IVP embryos. The CT method and collapsing the blastocoele prior to cryopreservation resulted in higher blastocyst survival rate. Further studies including transfer of embryos to recipients are necessary to corroborate these results.

O objetivo do presente estudo foi o de determinar em qual fase da onda de crescimento folicular são obtidas maiores taxas de recuperação e competência de oócitos obtidos por aspiração folicular (OPU) em novilhas cruzadas (Bos indicus x Bos taurus; Capítulo 1) ou em novilhas bubalinas (Bubalus bubalis), zebuínas (Nelore; Bos indicus) e taurinas (Holandês; Bos taurus; Capítulo 2). No Capítulo 1, 30 novilhas cruzadas foram submetidas à OPU aos 2 (D2), 4 (D4) e 6 (D6) dias após a emergência da onda de crescimento folicular (n=10/ grupo), em delineamento &quot;cross-over&quot; realizado em 3 réplicas. Os oócitos recuperados foram selecionados e os viáveis submetidos aos procedimentos de produção in vitro. Os dados foram analisados por ANOVA, utilizando o PROC MIXED, considerando os efeitos fixos de momento de OPU, de réplica e da interação entre eles, e o efeito aleatório de doadora. Foram produzidos maior taxa de blastocistos aos 6 dias pós-FIV, bem como maior número de blastocistos aos 9 dias pós-FIV e maior número de núcleos dos embriões eclodidos quando procedeu-se com aspiração no D2. Contudo, a OPU realizada no D6 também influenciou positivamente a taxa de blastocistos 6 dias pós FIV e não diferiu dos outros momentos com relação ao número de núcleos dos embriões eclodidos. No Capítulo 2, foram realizados 2 experimentos (1 e 2). No Experimento 1, foi avaliado o efeito de 50 mg de progesterona injetável na sincronização de emergência folicular em novilhas Nelore, Holandesas e bubalinas submetidas a protocolo de sincronização à base de progestágeno associado a 2 mg de benzoato de estradiol (n=10/ grupo). Os dados foram analisados por regressão logística, utilizando o PROC GLIMMIX. O momento da emergência folicular não foi afetado pela administração de progesterona injetável, ocorrendo entre 4,2 a 4,5 dias após o início do protocolo de sincronização para os três grupos genéticos. No Experimento 2, foram avaliados os efeitos de momento de sincronização da emergência folicular para a aspiração folicular (1, 3 ou 5 dias após a emergência) e de grupo genético (Nelore, Holandês e búfalo) na produção de embriões in vitro. Um total de 27 novilhas (9 de cada grupo genético, subdivididas nos 3 momentos de aspiração folicular) foram submetidas a 6 sessões de OPU, em delineamento &quot;cross-over&quot;. Os oócitos recuperados foram selecionados e os viáveis submetidos aos procedimentos de produção in vitro. Os dados foram analisados por ANOVA, utilizando o PROC MIXED, considerando os efeitos fixos de momento de OPU, de grupo genético, de réplica e da interação entre eles, e o efeito aleatório de doadora. Os resultados demonstraram que novilhas Nelore apresentaram melhor eficiência para a produção de embriões in vitro do que as novilhas Holandesas ou bubalinas, e que a aspiração folicular realizada em diferentes momentos da onda folicular sincronizada não influenciou a produção in vitro. / The objective of the present study was to determine in which phase of follicular wave higher recovery rates and better oocyte competence can be obtained by ovum pickup (OPU) in crossbred (Bos indicus x Bos taurus; Chapter 1) or buffalo (Bubalus bubalis), Nelore (Bos indicus) and Holstein heifers (Bos taurus; Chapter 2). In Chapter 1, 30 crossbred heifers were submitted to OPU on days 2 (D2), 4 (D4) or 6 (D6) after follicular wave emergence (n=10/ group), in a cross-over design performed in 3 replicates. Oocytes recovered were selected and those viable were submitted to in vitro embryo procedures. Data were analyzed by ANOVA, using PROC MIXED, in which time of OPU, replicate and interaction between these factors were considered as fixed effects, and donor was considered as a random effect. Higher blastocyst rate (6 days after IVF), number of blastocysts (9 days after IVF), and number of nuclei of hatched embryos were produced when OPU was done on D2. However, OPU on D6 also produced a higher blastocyst rate (6 days after IVF), and did not differ from other times of OPU concerning number of nuclei of hatched embryos. In Chapter 2, two experiments were performed. In Experiment 1, the effect of 50 mg of injectable progesterone was evaluated in the synchronization of follicular wave emergence of Nelore, Holstein, and buffalo heifers submitted to a progestin plus 2 mg of estradiol benzoate protocol (n=10/ group). Data were analyzed by logistic regression, by PROC GLIMMIX. The time of follicular wave emergence was not affected by administration of injectable progesterone. Besides, follicular wave emergence occurred between 4.2 and 4.5 days after the beginning of synchronization protocol for all genetic groups. In Experiment 2, the effects of time of OPU relative to follicular wave emergence (1, 3 or 5 days) and of genetic group (Nelore, Holstein, and buffalo) on in vitro embryo production were evaluated. A total of 27 heifers (9 of each genetic group, assigned in the 3 times of OPU) were submitted to 6 OPU sessions, in a cross-over design. Oocytes recovered were selected and those viable were submitted to in vitro embryo procedures. Data were analyzed by ANOVA, using PROC MIXED, in which time of OPU, genetic group, replicate and interaction between these factors were considered as fixed effects, and donor was considered as a random effect. Results demonstrated that Nelore heifers showed a better efficiency for in vitro embryo production than Holstein or buffalo heifers. Additionally, OPU performed at different times of synchronized follicular wave did not influence in vitro embryo production.

O objetivo do presente estudo foi o de determinar em qual fase da onda de crescimento folicular são obtidas maiores taxas de recuperação e competência de oócitos obtidos por aspiração folicular (OPU) em novilhas cruzadas (Bos indicus x Bos taurus; Capítulo 1) ou em novilhas bubalinas (Bubalus bubalis), zebuínas (Nelore; Bos indicus) e taurinas (Holandês; Bos taurus; Capítulo 2). No Capítulo 1, 30 novilhas cruzadas foram submetidas à OPU aos 2 (D2), 4 (D4) e 6 (D6) dias após a emergência da onda de crescimento folicular (n=10/ grupo), em delineamento &quot;cross-over&quot; realizado em 3 réplicas. Os oócitos recuperados foram selecionados e os viáveis submetidos aos procedimentos de produção in vitro. Os dados foram analisados por ANOVA, utilizando o PROC MIXED, considerando os efeitos fixos de momento de OPU, de réplica e da interação entre eles, e o efeito aleatório de doadora. Foram produzidos maior taxa de blastocistos aos 6 dias pós-FIV, bem como maior número de blastocistos aos 9 dias pós-FIV e maior número de núcleos dos embriões eclodidos quando procedeu-se com aspiração no D2. Contudo, a OPU realizada no D6 também influenciou positivamente a taxa de blastocistos 6 dias pós FIV e não diferiu dos outros momentos com relação ao número de núcleos dos embriões eclodidos. No Capítulo 2, foram realizados 2 experimentos (1 e 2). No Experimento 1, foi avaliado o efeito de 50 mg de progesterona injetável na sincronização de emergência folicular em novilhas Nelore, Holandesas e bubalinas submetidas a protocolo de sincronização à base de progestágeno associado a 2 mg de benzoato de estradiol (n=10/ grupo). Os dados foram analisados por regressão logística, utilizando o PROC GLIMMIX. O momento da emergência folicular não foi afetado pela administração de progesterona injetável, ocorrendo entre 4,2 a 4,5 dias após o início do protocolo de sincronização para os três grupos genéticos. No Experimento 2, foram avaliados os efeitos de momento de sincronização da emergência folicular para a aspiração folicular (1, 3 ou 5 dias após a emergência) e de grupo genético (Nelore, Holandês e búfalo) na produção de embriões in vitro. Um total de 27 novilhas (9 de cada grupo genético, subdivididas nos 3 momentos de aspiração folicular) foram submetidas a 6 sessões de OPU, em delineamento &quot;cross-over&quot;. Os oócitos recuperados foram selecionados e os viáveis submetidos aos procedimentos de produção in vitro. Os dados foram analisados por ANOVA, utilizando o PROC MIXED, considerando os efeitos fixos de momento de OPU, de grupo genético, de réplica e da interação entre eles, e o efeito aleatório de doadora. Os resultados demonstraram que novilhas Nelore apresentaram melhor eficiência para a produção de embriões in vitro do que as novilhas Holandesas ou bubalinas, e que a aspiração folicular realizada em diferentes momentos da onda folicular sincronizada não influenciou a produção in vitro. / The objective of the present study was to determine in which phase of follicular wave higher recovery rates and better oocyte competence can be obtained by ovum pickup (OPU) in crossbred (Bos indicus x Bos taurus; Chapter 1) or buffalo (Bubalus bubalis), Nelore (Bos indicus) and Holstein heifers (Bos taurus; Chapter 2). In Chapter 1, 30 crossbred heifers were submitted to OPU on days 2 (D2), 4 (D4) or 6 (D6) after follicular wave emergence (n=10/ group), in a cross-over design performed in 3 replicates. Oocytes recovered were selected and those viable were submitted to in vitro embryo procedures. Data were analyzed by ANOVA, using PROC MIXED, in which time of OPU, replicate and interaction between these factors were considered as fixed effects, and donor was considered as a random effect. Higher blastocyst rate (6 days after IVF), number of blastocysts (9 days after IVF), and number of nuclei of hatched embryos were produced when OPU was done on D2. However, OPU on D6 also produced a higher blastocyst rate (6 days after IVF), and did not differ from other times of OPU concerning number of nuclei of hatched embryos. In Chapter 2, two experiments were performed. In Experiment 1, the effect of 50 mg of injectable progesterone was evaluated in the synchronization of follicular wave emergence of Nelore, Holstein, and buffalo heifers submitted to a progestin plus 2 mg of estradiol benzoate protocol (n=10/ group). Data were analyzed by logistic regression, by PROC GLIMMIX. The time of follicular wave emergence was not affected by administration of injectable progesterone. Besides, follicular wave emergence occurred between 4.2 and 4.5 days after the beginning of synchronization protocol for all genetic groups. In Experiment 2, the effects of time of OPU relative to follicular wave emergence (1, 3 or 5 days) and of genetic group (Nelore, Holstein, and buffalo) on in vitro embryo production were evaluated. A total of 27 heifers (9 of each genetic group, assigned in the 3 times of OPU) were submitted to 6 OPU sessions, in a cross-over design. Oocytes recovered were selected and those viable were submitted to in vitro embryo procedures. Data were analyzed by ANOVA, using PROC MIXED, in which time of OPU, genetic group, replicate and interaction between these factors were considered as fixed effects, and donor was considered as a random effect. Results demonstrated that Nelore heifers showed a better efficiency for in vitro embryo production than Holstein or buffalo heifers. Additionally, OPU performed at different times of synchronized follicular wave did not influence in vitro embryo production.

Adaptive path planning of an autonomous marine vehicle (surface or subsurface) in the role of a communication and navigation aid (CNA) for multiple autonomous underwater vehicles (AUVs) for survey missions is studied. This path planning algorithm can be run before deployment, based on the planned paths of the survey AUVs, or underway, based on information transmitted by the survey AUVs. The planner considers the relative depth of the CNA and survey AUVs (not previously done) allowing the CNA to better aid survey AUVs that maintain a set distance over the ocean floor while surveying. Results are presented from simulations and in-water trials for both pre-deployment and underway planning modes, the latter being preferred since it can adapt to the survey AUV path during the mission. The necessity of bounding the distance between the CNA and any survey AUV in order to bound survey AUV position error is also described.

The purpose of this paper is to examine the role of logotypes within the visual identity and identify possible reasons that motivate companies to change their logo. This paper should also clarify how different parts of the visual identity affect each other and how they are interconnected. The thesis research question is formulated with the purpose in mind and has led to the following question: How can a logo change contribute to enhanced visual identity?   Since we wanted to obtain a better understanding of the chosen research area with help of interviews have we chosen a qualitative method for this thesis. During the process we have let the empirical material control the chosen theories. Our gathered theories are from secondary data and the empirical material has been collected from semi-structured interviews.  In order to obtain a wider range of our research topic, we have selected the interviewed respondents from different industries and areas of work. In our work process, we have analyzed and discussed the impact of a logo and how a logo change could impact on the visual identity. Our research showed that a single logo change would not necessarily have an impact on the visual identity but would improve with a combined match of the identities other components. During our process we also identified three main ingredients that a good logo should contain. Finally, we have developed a theory that shows how visual identity components are interconnected.