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Inúmeros aspectos da reprodução em Apis mellifera já foram extensamente divulgados, no entanto, os mecanismos reguladores da manutenção do estado estéril das operárias, bem como aqueles que permitem a ativação de seus ovários, ainda estão para serem descobertos. Por exemplo, a organização dos folículos ovarianos em crescimento e a arquitetura e papel das células foliculares neste processo. Além disso, para compreender o processo de ativação dos ovários em um contexto mais amplo, também é necessária uma investigação da síntese e maturação de diferentes classes de RNAs as quais modelam redes de interações gênicas extremamente complexas. Portanto, neste doutorado, tivemos como objetivo realizar 1- uma análise morfológica dos ovários ativos de operárias de A. mellifera obtidos em condições orfandade, com ênfase nas células foliculares e 2- um estudo aprofundado da regulação da expressão gênica (genes estruturais e reguladores) que é de fundamental importância para ligar os genótipos aos fenótipos. A análise morfológica dos ovários de operárias de A. mellifera foi realizada em microscópio de fluorescência ou confocal (priorizou a contagem das células foliculares) e microscópio eletrônico de transmissão, que permitiu a descrição e caracterização, pela primeira vez, da patência em ovários de operárias A. mellifera. Paralelamente, por meio da técnica de RNAseq, foi possível analisar o transcriptoma (miRNAs e mRNAs) de amostras específicas de ovários, em diferentes estados fisiológicos, em rainhas e operárias. Os mRNAs e miRNAs que se destacaram em nossas análises in silico foram validados experimentalmente por RT-PCR com alto grau de reprodutibilidade e em harmonia com o estado fisiológico dos ovários. Os transcritos altamente expressos nos ovários ativados foram: fpps5, cad, obp7, yellow-g e aqueles representados pelo GB42182 e GB44975. Acreditamos estes genes possam fazer parte da rede que regula o processo de ativação dos ovários em A. mellifera. Os miRNAs que se destacaram em nossas análises foram: A) miR-306 e miR-317 - altamente expressos nas amostras de ovários funcionais e B) miR-71 pelo fato de, nas análises in silico, ser o mais forte candidato a alvejar a vitelogenina, e na análise experimental, apresentarem, microRNAs e mRNAs, perfis de expressão antagônicos. A construção de bibliotecas de microRNAs e mRNAs a partir de ovários funcionais e não funcionais de abelhas operárias e rainhas, a análise de expressão, bem como a predição de uma rede de integração nos deu um retrato do sensível equilíbrio reprodutivo que mantém ambas as castas em aparente harmonia dentro da colônia aonde elas assumem, no momento certo, seus papéis nesta sofisticada sociedade empreendendo ou não a reprodução. / Countless aspects of reproduction in Apis mellifera have been widely published, however, the regulatory mechanisms for the maintenance of the sterile state of workers as well as those that allow the activation of their ovaries are still to be discovered, as much as the organization of growing ovarian follicles, the architecture and the role of follicular cells during this process. Furthermore, to understand the activation process of the ovaries in a broader context, it is also necessary to investigate the synthesis and maturation of different classes of RNAs which exemplify networks of gene interactions, extremely complex. Therefore, PhD project, we aimed to approach: 1 - A morphological analysis of active ovaries of A. mellifera workers obtained in queenless conditions, with emphasis on the follicular cells and 2 - A detailed study of the regulation of gene expression (structural and regulatory genes) that is crucial for linking genotypes to phenotypes. Morphologic analysis of workers ovaries of A. mellifera was performed under a fluorescence microscope or confocal (prioritized follicular cell count) and transmission electron microscope, which allowed, for the first time, a description and characterization of the patency of worker ovaries in A. mellifera. Similarly, by RNAseq technique, it was possible to analyze the transcriptome (miRNAs and mRNAs) of specific samples of ovaries at different physiological states, in queens and workers. mRNAs and miRNAs that stood out in our in silico analysis were experimentally validated by RT-PCR with high reproducibility and in harmony with ovaries physiologic state. Transcripts highly expressed in activated ovaries were fpps5, cad, obp7, yellow-g and those represented by GB42182 and GB44975. We believe these genes may be part of the network that regulates ovaries activation process in A. mellifera. miRNAs that stood out in our analysis were: - a) - miR-306 and miR-317 - highly expressed in samples of active ovaries and b) -miR-71 by the fact that the in silico analysis, was the strongest candidate to target vitellogenin, and in experimental analysis, presented antagonistic profile of expression when microRNAs and mRNAs were contrasted. The construction of microRNAs and mRNAs libraries from active and inactive ovaries of worker bees and queens, the analysis expression, as well as the prediction of a integrative network has given us a portrait of the sensitive reproductive balance that keeps both castes of bees in apparent harmony within the colony, where they take each one, at the right time, their roles in this sophisticated society, undertaking or not the reproduction.

O fator tuboperitoneal é a causa mais freqüente de infertilidade na mulher e a sua investigação é fundamental na propedêutica do casal infértil. Neste estudo, avaliou-se o método da sonohisterossalpingografia contrastada (Hycosy), que utiliza o contraste SH U 454 (Echovist), na detecção da permeabilidade tubária, comparando-o com a histerossalpingografia (HSG) e com a laparoscopia (LPS) acompanhada de cromotubagem (padrão ouro). Realizou-se a investigação de 96 tubas uterinas de 48 pacientes, submetidas à Hycosy, HSG e LPS. Todas as avaliações da Hycosy foram realizadas por um único observador que desconhecia o resultado da HSG. O mesmo se refere, também, quanto à realização da HSG, bem como da LPS. Foram avaliados os seguintes instrumentos de medidas: sensibilidade (S), especificidade (E), valor preditivo positivo (VPP), valor preditivo negativo (VPN), concordância observada (CO), razão de verossimilhança positiva (RV+) e razão de verossimilhança negativa (RV-). Comparando-se a avaliação pela Hycosy com a LPS, foram encontrados os seguintes resultados: S=86,6%, E=35,7%, VPP=88,75%, VPN=31,25%, CO=79,2%, RV+=1,35 e RV-=0,38. Quanto à comparação da HSG com a LPS, os valores obtidos foram: S=85,4%, E= 50%, VPP=90,9%, VPN=36,8%, CO=80,2%, RV+=1,71 e RV-=0,29. Entre Hycosy e HSG, a co-positividade foi de 87%, a co-negatividade de 31,6% e a concordância de 76%. Esse estudo demonstrou que a Hycosy é uma técnica segura e confiável para a avaliação da permeabilidade tubária na propedêutica da infertilidade. / Tubal disorders are the main causes of female infertility. Assessment of tubal patency is a very important issue for evaluating an infertile couple. A total of 96 Fallopian tubes from 48 patients was examined with hysterosalpingo-contrast sonography (Hycosy), hysterosalpingography (HSG) and laparoscopic chromopertubation (LPS). Hycosy was performed by transvaginal ultrasound with a new contrast medium (Echovist). Compared with laparoscopic results, we found a sensitivity of 86.6%, a specificity of 35.7%, positive predictive value (VPP) of 88.75%, negative predictive value (VPN) of 31.25% and concordance (Hycosy/LPS) of 79,2%, while the results for HSG were sensitivity of 85.4%, specificity of 50%, VPP of 90.9%, VPN of 36.8% and concordance (HSG/LPS) of 80.2%. In conclusion, the results have shown that Hycosy is a reliable and safe method for evaluating tubal patency.

<p>The primary aim of this thesis was to modify saphenous vein (SV) harvesting technique and evaluate its clinical importance. A new "no touch" (NT) technique of SV preparation was developed where the vein is harvested with a pedicle of surrounding tissue, which protects the vein from spasm therefore obviating the need for distension.</p><p>Firstly, a prospective randomised study in 156 patients who underwent coronary artery bypass grafting was done to compare this new "no touch" technique to two others, the conventional (C) and the intermediate (I) techniques. A morphological study of the endothelium showed an endothelial integrity of 97% in NT vessels while about half of endothelial surface of veins harvested by the other two techniques was devoid of endothelium. At angiographic follow up, the patency for NT was 95.4%, 88.9% for grafts in group C and 86.2% for grafts in group I. A statistically significant difference in patency rate was found between the NT group and group C (p=0.025) and the poorest result was observed in group I.</p><p>Secondly, the immunohistochemistry assessment using CD31-antibody confirmed a better-preserved endothelium for NT vessels. Putative NOS was identified by NADPH-diaphorase histochemistry and autoradiographic localization of [3H] L-nitroarginine (NOARG) binding. NADPH staining was almost continuous on the luminal aspect and was also present in the intact adventitia of NT vessels, which was markedly reduced in conventionally harvested veins. Autoradiographic analysis of specific NOARG binding showed greater binding in the no-touch vessels, confirming the histochemistry results. All three NOS isoforms were identified in the media of SV grafts. In NT, NOS I was abundant in adventitial nerves; NOS II was found in adventitial vasa vasorum and NOS III was associated with endothelial cells lining both the vessel lumen and microvessels within the adventitia.</p><p>In conclusion, this study demonstrated that the endothelial integrity and NOS activity are better maintained when using the no-touch technique for vein graft harvesting. The vasorelaxant and thromboresistent activities of NO may be responsible for the reduced venospasm and improved early patency rates observed. Furthermore, the mechanical properties provided by the cushion of surrounding tissue in grafts harvested by NT technique may contribute to the observed high patency rate.</p>

The primary aim of this thesis was to modify saphenous vein (SV) harvesting technique and evaluate its clinical importance. A new "no touch" (NT) technique of SV preparation was developed where the vein is harvested with a pedicle of surrounding tissue, which protects the vein from spasm therefore obviating the need for distension. Firstly, a prospective randomised study in 156 patients who underwent coronary artery bypass grafting was done to compare this new "no touch" technique to two others, the conventional (C) and the intermediate (I) techniques. A morphological study of the endothelium showed an endothelial integrity of 97% in NT vessels while about half of endothelial surface of veins harvested by the other two techniques was devoid of endothelium. At angiographic follow up, the patency for NT was 95.4%, 88.9% for grafts in group C and 86.2% for grafts in group I. A statistically significant difference in patency rate was found between the NT group and group C (p=0.025) and the poorest result was observed in group I. Secondly, the immunohistochemistry assessment using CD31-antibody confirmed a better-preserved endothelium for NT vessels. Putative NOS was identified by NADPH-diaphorase histochemistry and autoradiographic localization of [3H] L-nitroarginine (NOARG) binding. NADPH staining was almost continuous on the luminal aspect and was also present in the intact adventitia of NT vessels, which was markedly reduced in conventionally harvested veins. Autoradiographic analysis of specific NOARG binding showed greater binding in the no-touch vessels, confirming the histochemistry results. All three NOS isoforms were identified in the media of SV grafts. In NT, NOS I was abundant in adventitial nerves; NOS II was found in adventitial vasa vasorum and NOS III was associated with endothelial cells lining both the vessel lumen and microvessels within the adventitia. In conclusion, this study demonstrated that the endothelial integrity and NOS activity are better maintained when using the no-touch technique for vein graft harvesting. The vasorelaxant and thromboresistent activities of NO may be responsible for the reduced venospasm and improved early patency rates observed. Furthermore, the mechanical properties provided by the cushion of surrounding tissue in grafts harvested by NT technique may contribute to the observed high patency rate.

Abstract Title: When the cat is away… A study of pedagogical leadership at a distance Level: Final assignment for Degree of Master of science (one year) in Educational Management Author: Evelyn Joelsson Supervisor: Ulla Sebrant Date: 2013 may Leadership is a subject that has been discussed for many generations. Today’s leadership is something completely different than what has been practiced before, in the more hierarchical companies, when leadership was something you were born to do and the workers had no choice but to follow. In the globalised world that we live in today there are demands of a different kind than before and physical distance between leaders and their followers gets more and more common. Communication is the focus and the catchwords are patency, trust and availability. In our information society it is important to have an open dialog and to aim for continuous development. Often this is built with close and supportive leadership. My question is; how can you be a close and supportive leader when you are physically somewhere else? To find the answer to my questions I began with studying some previous work that has been done in this area, Maria Nordengren &amp; Bengt Olsen (2006) among others. I interviewed ten people who all have some kind of experience with working at a distance and the professional leadership from afar. Two of them were leaders, four were employees that worked at a distance from the leader and four were employees working physically close to the leader. I asked them about communication, trust, patency and availability and found out that educational leadership is possible at a distance but also at a price. If you are a leader placed at a distance from your employees you need to be clearer in your communication than the leader close at hand and give the right information at the right time. Trust is very important as well as the physical meetings they have twice a year. Availability is important and many claimed that it was working satisfactory. The physical contact proved to be irreplaceable even if there are many technical tools available in today’s businesses. Employees at a physical distance from their leaders automatically need to take more decisions on their own and learn to take initiative faster than the ones who can just walk over to the leader and ask. For future research I suggest that you could look at the different kinds of distance between leaders and their followers, not just de physical, and maybe also the gender aspect of leadership at a distance. One could conduct a comparative study between different kinds of companies and see if IT-businesses differ from producing- or service companies. The communication strategies that were used was something I found interesting and is probably something you can look more into, for example when you use what tool for communication. I think my thesis has contributed to the understanding of the complexity of leadership at a physical distance and what it takes to be an educational leader in a scattered company today concerning communication, patency, trust and availability. Keywords: leadership, educational leadership, leadership at a distance, communication, distance, development opportunity, trust, patency and availability.

The role of dopamine in directly modulating somatic motoneuron excitability and hence muscle tone is unknown. We investigated whether dopamine influences the trigeminal motor pool (MoV) that innervates the masseter and tensor palatini muscles, both of which function to maintain upper airway patency. We hypothesized that dopamine facilitates motor outflow at the MoV. We focally applied apomorphine (nonspecific dopamine receptor agonist) at the MoV in anaesthetized rats. We also applied receptor-specific agonists and antagonists to determine the receptor subtype mediating dopaminergic mechanisms of action. We demonstrated that dopaminergic transmission at the MoV potently increased motor outflow via the D1-like receptor and facilitated masseter and tensor palatini muscle tone. It is unknown whether endogenous dopamine release on to airway motoneurons influences their activity to regulate muscle tone in natural sleep-wake behaviours. This issue warrants investigation because the neurochemical basis of upper airway motor dysfunction (e.g. obstructive sleep apnea) remains poorly characterized.

The role of dopamine in directly modulating somatic motoneuron excitability and hence muscle tone is unknown. We investigated whether dopamine influences the trigeminal motor pool (MoV) that innervates the masseter and tensor palatini muscles, both of which function to maintain upper airway patency. We hypothesized that dopamine facilitates motor outflow at the MoV. We focally applied apomorphine (nonspecific dopamine receptor agonist) at the MoV in anaesthetized rats. We also applied receptor-specific agonists and antagonists to determine the receptor subtype mediating dopaminergic mechanisms of action. We demonstrated that dopaminergic transmission at the MoV potently increased motor outflow via the D1-like receptor and facilitated masseter and tensor palatini muscle tone. It is unknown whether endogenous dopamine release on to airway motoneurons influences their activity to regulate muscle tone in natural sleep-wake behaviours. This issue warrants investigation because the neurochemical basis of upper airway motor dysfunction (e.g. obstructive sleep apnea) remains poorly characterized.

Inúmeros aspectos da reprodução em Apis mellifera já foram extensamente divulgados, no entanto, os mecanismos reguladores da manutenção do estado estéril das operárias, bem como aqueles que permitem a ativação de seus ovários, ainda estão para serem descobertos. Por exemplo, a organização dos folículos ovarianos em crescimento e a arquitetura e papel das células foliculares neste processo. Além disso, para compreender o processo de ativação dos ovários em um contexto mais amplo, também é necessária uma investigação da síntese e maturação de diferentes classes de RNAs as quais modelam redes de interações gênicas extremamente complexas. Portanto, neste doutorado, tivemos como objetivo realizar 1- uma análise morfológica dos ovários ativos de operárias de A. mellifera obtidos em condições orfandade, com ênfase nas células foliculares e 2- um estudo aprofundado da regulação da expressão gênica (genes estruturais e reguladores) que é de fundamental importância para ligar os genótipos aos fenótipos. A análise morfológica dos ovários de operárias de A. mellifera foi realizada em microscópio de fluorescência ou confocal (priorizou a contagem das células foliculares) e microscópio eletrônico de transmissão, que permitiu a descrição e caracterização, pela primeira vez, da patência em ovários de operárias A. mellifera. Paralelamente, por meio da técnica de RNAseq, foi possível analisar o transcriptoma (miRNAs e mRNAs) de amostras específicas de ovários, em diferentes estados fisiológicos, em rainhas e operárias. Os mRNAs e miRNAs que se destacaram em nossas análises in silico foram validados experimentalmente por RT-PCR com alto grau de reprodutibilidade e em harmonia com o estado fisiológico dos ovários. Os transcritos altamente expressos nos ovários ativados foram: fpps5, cad, obp7, yellow-g e aqueles representados pelo GB42182 e GB44975. Acreditamos estes genes possam fazer parte da rede que regula o processo de ativação dos ovários em A. mellifera. Os miRNAs que se destacaram em nossas análises foram: A) miR-306 e miR-317 - altamente expressos nas amostras de ovários funcionais e B) miR-71 pelo fato de, nas análises in silico, ser o mais forte candidato a alvejar a vitelogenina, e na análise experimental, apresentarem, microRNAs e mRNAs, perfis de expressão antagônicos. A construção de bibliotecas de microRNAs e mRNAs a partir de ovários funcionais e não funcionais de abelhas operárias e rainhas, a análise de expressão, bem como a predição de uma rede de integração nos deu um retrato do sensível equilíbrio reprodutivo que mantém ambas as castas em aparente harmonia dentro da colônia aonde elas assumem, no momento certo, seus papéis nesta sofisticada sociedade empreendendo ou não a reprodução. / Countless aspects of reproduction in Apis mellifera have been widely published, however, the regulatory mechanisms for the maintenance of the sterile state of workers as well as those that allow the activation of their ovaries are still to be discovered, as much as the organization of growing ovarian follicles, the architecture and the role of follicular cells during this process. Furthermore, to understand the activation process of the ovaries in a broader context, it is also necessary to investigate the synthesis and maturation of different classes of RNAs which exemplify networks of gene interactions, extremely complex. Therefore, PhD project, we aimed to approach: 1 - A morphological analysis of active ovaries of A. mellifera workers obtained in queenless conditions, with emphasis on the follicular cells and 2 - A detailed study of the regulation of gene expression (structural and regulatory genes) that is crucial for linking genotypes to phenotypes. Morphologic analysis of workers ovaries of A. mellifera was performed under a fluorescence microscope or confocal (prioritized follicular cell count) and transmission electron microscope, which allowed, for the first time, a description and characterization of the patency of worker ovaries in A. mellifera. Similarly, by RNAseq technique, it was possible to analyze the transcriptome (miRNAs and mRNAs) of specific samples of ovaries at different physiological states, in queens and workers. mRNAs and miRNAs that stood out in our in silico analysis were experimentally validated by RT-PCR with high reproducibility and in harmony with ovaries physiologic state. Transcripts highly expressed in activated ovaries were fpps5, cad, obp7, yellow-g and those represented by GB42182 and GB44975. We believe these genes may be part of the network that regulates ovaries activation process in A. mellifera. miRNAs that stood out in our analysis were: - a) - miR-306 and miR-317 - highly expressed in samples of active ovaries and b) -miR-71 by the fact that the in silico analysis, was the strongest candidate to target vitellogenin, and in experimental analysis, presented antagonistic profile of expression when microRNAs and mRNAs were contrasted. The construction of microRNAs and mRNAs libraries from active and inactive ovaries of worker bees and queens, the analysis expression, as well as the prediction of a integrative network has given us a portrait of the sensitive reproductive balance that keeps both castes of bees in apparent harmony within the colony, where they take each one, at the right time, their roles in this sophisticated society, undertaking or not the reproduction.

O fator tuboperitoneal é a causa mais freqüente de infertilidade na mulher e a sua investigação é fundamental na propedêutica do casal infértil. Neste estudo, avaliou-se o método da sonohisterossalpingografia contrastada (Hycosy), que utiliza o contraste SH U 454 (Echovist&#61650;), na detecção da permeabilidade tubária, comparando-o com a histerossalpingografia (HSG) e com a laparoscopia (LPS) acompanhada de cromotubagem (padrão ouro). Realizou-se a investigação de 96 tubas uterinas de 48 pacientes, submetidas à Hycosy, HSG e LPS. Todas as avaliações da Hycosy foram realizadas por um único observador que desconhecia o resultado da HSG. O mesmo se refere, também, quanto à realização da HSG, bem como da LPS. Foram avaliados os seguintes instrumentos de medidas: sensibilidade (S), especificidade (E), valor preditivo positivo (VPP), valor preditivo negativo (VPN), concordância observada (CO), razão de verossimilhança positiva (RV+) e razão de verossimilhança negativa (RV-). Comparando-se a avaliação pela Hycosy com a LPS, foram encontrados os seguintes resultados: S=86,6%, E=35,7%, VPP=88,75%, VPN=31,25%, CO=79,2%, RV+=1,35 e RV-=0,38. Quanto à comparação da HSG com a LPS, os valores obtidos foram: S=85,4%, E= 50%, VPP=90,9%, VPN=36,8%, CO=80,2%, RV+=1,71 e RV-=0,29. Entre Hycosy e HSG, a co-positividade foi de 87%, a co-negatividade de 31,6% e a concordância de 76%. Esse estudo demonstrou que a Hycosy é uma técnica segura e confiável para a avaliação da permeabilidade tubária na propedêutica da infertilidade. / Tubal disorders are the main causes of female infertility. Assessment of tubal patency is a very important issue for evaluating an infertile couple. A total of 96 Fallopian tubes from 48 patients was examined with hysterosalpingo-contrast sonography (Hycosy), hysterosalpingography (HSG) and laparoscopic chromopertubation (LPS). Hycosy was performed by transvaginal ultrasound with a new contrast medium (Echovist&#61650;). Compared with laparoscopic results, we found a sensitivity of 86.6%, a specificity of 35.7%, positive predictive value (VPP) of 88.75%, negative predictive value (VPN) of 31.25% and concordance (Hycosy/LPS) of 79,2%, while the results for HSG were sensitivity of 85.4%, specificity of 50%, VPP of 90.9%, VPN of 36.8% and concordance (HSG/LPS) of 80.2%. In conclusion, the results have shown that Hycosy is a reliable and safe method for evaluating tubal patency.