Thèses : « Human Profile »

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Devaraj, Revathy. « Validation of the Human Activity Profile ». Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp01/MQ52893.pdf.

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Burden, Helena P. « Prostate cell profile in human male urine ». Thesis, University of Bristol, 2010. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.539764.

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Trudel, Nathalie. « Gene expression profile in human prostate cancer cell lines ». Thesis, McGill University, 2000. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=33449.

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Since the beginning of this work in 1998, it is estimated that 1780 men died of prostate cancer in Quebec. Molecular analysis of prostate cancer will eventually lead to the discovery of key genes involved in its onset and progression. The present project was to compare gene expression profiles in human non-tumorigenic versus tumorigenic prostate cell lines generated in our laboratory. A putative tumor suppressor gene present on 12q13 would be responsible for the non-tumorigenic phenotype of one cell line as discovered earlier by our team.
In order to compare gene expression patterns, expression arrays from Clontech, bearing 588 genes known to be involved in human cancers, were hybridized with cDNA derived from two related cell lines available in our laboratory. This one experiment provided interesting hints on differentially expressed genes that could be involved in human prostate cancer. Interesting clones were confirmed by Northern blots. When commercial antibody was available, analysis was extended at the protein level. A combination of these analyses revealed no striking difference in the level of expression for the genes previously identified by the arrays hybridization.
Simultaneously, differential display PCR techniques, allowing the discovery of unknown differentially expressed molecules and thus complementing the previous approach, were applied to compare related cell lines and unique hybrids. Cloning and sequencing of differential fragments brought us to what could be a new cDNA expressed in many human cell lines.
Prostate cancer is not well characterized enough to allow accurate diagnosis or appropriate therapy strategies. Differentially expressed molecules analyzed in this project as well as the putative new cDNA might fulfil part of this lack in the understanding of this disease.

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Feng, Huichen. « Gene expression profile in human trophoblast and gestational trophoblastic disease ». Click to view the E-thesis via HKUTO, 2004. http://sunzi.lib.hku.hk/hkuto/record/B31384742.

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Warasiha, Benjamart. « Cytochrome P450 mRNA profile in human breast cancer cell lines ». Thesis, Robert Gordon University, 2008. http://hdl.handle.net/10059/364.

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Cytochrome P450 enzymes (P450s) are involved in cancer development and treatment due to their roles in the oxidative metabolism of various endogenous (e.g. oestrogen) and exogenous (e.g. tamoxifen) compounds. It is well-known that intermediate P450 metabolites derived from oestrogen metabolism are associated with breast carcinogenesis. The main aim of this project was to profile the cytochrome P450 and P450-regulatory nuclear receptor mRNAs in a series of breast cancer cell lines (BCCs) and compare this profile with normal breast cells. This study used the qualitative reverse transcriptasepolymerase chain reaction (RT-PCR) to detect mRNA expression of target genes. Results showed CYP1B1, CYP2D6, CYP2J2, CYP2R1, CYP2U1 and CYP4X1 mRNA to be present in all cell lines. CYP2A6, CYP2C8, CYP2C18, CYP2F1 and CYP4Z1 mRNA were expressed in oestrogen receptor (ER)-positiveCaucasian and ER-negative Afro- Caribbean BCCs. Although no differences in P450 mRNA were observed between the different ethnic groups, these preliminary findings suggest potential similarities in the ERpositive Caucasian and ER-negative Afro-Caribbean BCCs which warrant further investigation The CYP4Z1 PCR product was identified as two distinct bands. Specific primer sets were used to demonstrate potential intron retention in CYP4Z1. Using established in vitro models for the study of regulatory mechanisms of CYP4Z1, T47D and ZR-75-1 breast cancer cell lines were used to determine the appropriate nuclear receptors (i.e. progesterone receptor, glucocorticoid receptor or peroxisome proliferator-activated receptor alpha ). These findings suggest that there may be an alternative receptor mechanism involved in CYP4Z1 mRNA induction in these cells. In conjunction, pre-treatment of these two cell lines with the RNA synthesis inhibitor actinomycin D followed by the agonists showed a significant reduction (p < 0.05) of CYP4Z1 mRNA levels and inhibited CYP4Z1 induction by either progesterone, dexamethasone or pirinixic acid, indicating that these agonists have effects on CYP4Z1 mRNA transcription or stability. In contrast, cycloheximide differentially affected the level of CYP4Z1 mRNA induction by these agonists. Taken together, these results suggest that CYP4Z1 mRNA induction in T47D and ZR-75-1 is mediated through differential cell type specific regulatory mechanisms and there is evidence for differential regulation of the splice variants.

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Feng, Huichen, et 馮會臣. « Gene expression profile in human trophoblast and gestational trophoblastic disease ». Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2004. http://hub.hku.hk/bib/B31384742.

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Chang, Yung Ping. « Gesture Analysis for Human-Computer Interface Using Profile-Matching Stereo Vision ». BYU ScholarsArchive, 2013. https://scholarsarchive.byu.edu/etd/3729.

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This thesis presents a novel profile shape matching stereo vision algorithm. This algorithm is able to obtain 3D information in real time from a pair of stereo images. This algorithm produces the 3D information by matching the profile intensity shapes on the same row of the two images from a stereo image pair. The advantage of this profile shape matching algorithm is that the detection of correspondences relies on intensity profile shape not on intensity values, which subject to lighting variations. The user can choose an interval of disparity, and then an object in a desired distance range can be segmented out from the background. In other words, the algorithm detects the object according to its distance to the cameras. Based on the resulting 3D information, the movement and gesture of the control agents, in our test cases the human body and fingers, in space in a desired distance range can be determined. The body movement and gestures can then be analyzed for human-computer interface purposes. In this thesis, the algorithm was applied for human pose and hand gesture estimation. To demonstrate its performance the estimation results were interpreted as inputs and sent to a smart phone to control its functions. While this algorithm does have a trade-off between accuracy and processing speed, we found a balance that can produce the result in real time, and the result has sufficient accuracy for practical use of recognizing human poses and hand gesture. The experimental result shows that the proposed algorithm has higher accuracy and is 1.14× faster than the original version on tested stereo image pairs.

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Arney, Christle Shannon. « Morphological variation in the human tibia and its potential for profile estimation in human skeletal remains ». Thesis, Wichita State University, 2011. http://hdl.handle.net/10057/5159.

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This study evaluates the existence and degree of human variation as it is represented by the morphology of the tibia. Specifically, this research is undertaken in order to quantitatively address the morphological variation of this skeletal element to reveal the inherent variation within the individual, while also evaluating the discrepancies that result due to the sex and age of an individual. It also explores the interaction of tibial morphology with living stature, assessing the ability of the quantified portions of the bone to explain stature. In order to investigate this variation, the tibiae of 382 mature skeletal remains from the Hamann-Todd Osteological Collection in Cleveland, Ohio, were analyzed. These specimens were comprised of 180 females and 202 males whose group affiliation were designated as American Black. Using univariate and multivariate analyses, the morphological variation of the human tibia was assessed in respect to goals outlined above. These analyses revealed the manner by which dimensions of the tibia covaries, providing a better understanding of the innate variation that exists within this bone. These procedures also enabled the evaluation of sex and age effects on the size and shape of the tibia, revealing that the variations due to sex are profound enough to allow accurate classification of the sexes from the morphology of the tibia. While age related changes impact the morphology of the bone, they do not impede the ability of the dimensions to be used as reliable sex indicators. Further, the assessment of the interaction of the tibia and stature demonstrates the degree by which the variables explain the stature variation in the sample, attesting to the capacity of the tibial dimensions to be used as predictors of stature. Finally, the efficacy of particular measurements employed throughout this study to obtain accurate information concerning human variation is established, as well as their applicability to fragmentary remains.
Thesis (M.A.)--Wichita State University, College of Liberal Arts and Sciences, Dept. of Anthropology.

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Harrington, Carl R. « Design and application of a DNA microarray to profile human gut flora ». Thesis, University of East Anglia, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.435018.

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Popova, I. S. « Usage of immunohistochemiсal markers for complex diagnosing of human breast cancer profile ». Thesis, БДМУ, 2021. http://dspace.bsmu.edu.ua:8080/xmlui/handle/123456789/18464.

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Nilsson, Felix. « Joint Human-Machine Exploration of Industrial Time Series Using the Matrix Profile ». Thesis, Högskolan i Halmstad, CAISR Centrum för tillämpade intelligenta system (IS-lab), 2021. http://urn.kb.se/resolve?urn=urn:nbn:se:hh:diva-44717.

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Technological advancements and widespread adaptation of new technology in industry have made industrial time series data more available than ever before. This trend is expected to continue, especially with the introduction of Industry 4.0, where the goal is to connect everything on the industry floor to the cloud and the Industrial Internet of Things. With this development grows the need for versatile methods for mining industrial time series data. Time series motif discovery is a sub-set of data mining and is about finding interesting patterns in time series data. The state of the art in time series motif discovery is the Matrix Profile proposed in 2016. However, there are few publications where the Matrix Profile has been applied to real-life industrial time series data despite its popularity. The goal of the thesis has been to create a tool that enables joint human-machine exploration of industrial time series data using the Matrix profile and present the challenges involved. The result is a human-machine exploration procedure called IUSE that has been applied to three data sets containing real-life industrial time series data. IUSE enables the user to extract semantic information, detect cycles, find deviating patterns and helps the user to get a deeper understanding of a time series. The description of IUSE comes alongside learned lessons, faced challenges and experience from applying the Matrix Profile to actual industrial time series data.

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Clark, Christopher Anderson. « Retinal profile and structural differences between myopes and emmetropes ». Thesis, Indiana University, 2014. http://pqdtopen.proquest.com/#viewpdf?dispub=3616989.

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Refractive development has been shown to be influenced by optical defocus in the eye and the interpretation of this signal appears to be localized in the retina. Optical defocus is not uniform across the retina and has been suggested as a potential cause of myopia development. Specifically hyperopic focus, i.e. focusing light behind the retina, may signal the eye to elongate, causing myopia. This non-uniform hyperopic signal appears to be due to the retinal shape. Ultimately, these signals are detected by the retina in an as yet undetermined manner. The purpose of this thesis is to examine the retinal profile using a novel method developed at Indiana University and then to examine retinal structural changes across the retina associated with myopia.

Myopes exhibited more prolate retinas than hyperopes/emmetropes using the SD OCT. Using the SD OCT, this profile difference was detectable starting at 5 degrees from the fovea, which was closer than previously reported in the literature. These results agreed significantly with results found from peripheral refraction and peripheral axial length at 10 degrees. Overall, the total retina was thinner for myopes than hyperopes/emmetropes. It was also statistically significantly thinner for the Outer Nuclear Layer (ONL), Inner Nuclear Layer (INL) and Outer Plexiform Layer (OPL) but not for other retinal layers such as the Ganglion Layer. Thinning generally occurred outside of 5 degrees.

The SD OCT method provided a nearly 10 fold increase in sensitivity which allowed for detection of profile changes closer to the fovea. The location of the retinal changes may be interesting as the layers that showed significant differences in thickness are also layers that contain cells believed to be associated with refractive development (amacrine, bipolar, and photoreceptor cells.) The reason for the retinal changes cannot be determined with this study, but possible theories include stretch due to axial elongation, neural remodeling due to blur, and/or direct influence on refractive development due to neural cell densities.

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Zschaler, Josefin, Juliane Dorow, Louisa Schöpe, Uta Ceglarek et Jürgen Arnhold. « Impact of Myeloperoxidase-derived oxidants on the product profile of human 5-Lipoxygenase ». Universitätsbibliothek Leipzig, 2016. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-201824.

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Human 5-lipoxygenase (5-LOX) oxidizes arachidonic acid to 5S-hydroperoxy-6E,8Z,11Z,14Z-eicosatetraenoic acid (5-HpETE) and leukotriene (LT) A4. In neutrophils, LTA4 is further converted to the potent chemoattractant LTB4. These cells also contain the heme enzyme myeloperoxidase (MPO), which produces several potent oxidants such as hypochlorous acid (HOCl), which are involved in pathogen defense and immune regulation. Here, we addressed the question whether MPO-derived oxidants are able to affect the activity of 5-LOX and the product profile of this enzyme. Human 5-LOX was incubated with increasing amounts of HOCl or HOBr. Afterward, arachidonic acid metabolites of 5-LOX were analyzed by reverse-phase high-performance liquid chromatography as well as by liquid chromatography-electrospray ionization-tandem mass spectrometry. The incubation of 5-LOX with the MPO-derived oxidants significantly changed the product profile of 5-LOX. Thereby, HOCl and HOBr increased the ratio of 5-H(p)ETE to 6-trans-LTB4 in a concentration-dependent manner. At low oxidant concentrations, there was a strong decrease in the yield of 6-trans-LTB4, whereas 5-HpETE did not change or increased. Additionally, the formation of 8-HpETE and 12-HpETE by 5-LOX rose slightly with increasing HOCl and HOBr. Comparable results were obtained with the MPO-H2O2-Cl– system when glucose oxidase and glucose were applied as a source of H2O2. This was necessary because of a strong impairment of 5-LOX activity by H2O2. In summary, MPO-derived oxidants showed a considerable impact on 5-LOX, impairing the epoxidation of 5-HpETE, whereas the hydroperoxidation of arachidonic acid was unaffected. Apparently, this was caused by an oxidative modification of critical amino acid residues of 5-LOX. Further work is necessary to assess the specific type and position of oxidation in the substrate-binding cavity of 5-LOX and to specify whether this interaction between 5-LOX and MPO-derived oxidants also takes place in stimulated neutrophils.

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Zschaler, Josefin, Juliane Dorow, Louisa Schöpe, Uta Ceglarek et Jürgen Arnhold. « Impact of Myeloperoxidase-derived oxidants on the product profile of human 5-Lipoxygenase ». Free radical biology & ; medicine (2015) 85, S. 148-156, 2015. https://ul.qucosa.de/id/qucosa%3A14677.

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Human 5-lipoxygenase (5-LOX) oxidizes arachidonic acid to 5S-hydroperoxy-6E,8Z,11Z,14Z-eicosatetraenoic acid (5-HpETE) and leukotriene (LT) A4. In neutrophils, LTA4 is further converted to the potent chemoattractant LTB4. These cells also contain the heme enzyme myeloperoxidase (MPO), which produces several potent oxidants such as hypochlorous acid (HOCl), which are involved in pathogen defense and immune regulation. Here, we addressed the question whether MPO-derived oxidants are able to affect the activity of 5-LOX and the product profile of this enzyme. Human 5-LOX was incubated with increasing amounts of HOCl or HOBr. Afterward, arachidonic acid metabolites of 5-LOX were analyzed by reverse-phase high-performance liquid chromatography as well as by liquid chromatography-electrospray ionization-tandem mass spectrometry. The incubation of 5-LOX with the MPO-derived oxidants significantly changed the product profile of 5-LOX. Thereby, HOCl and HOBr increased the ratio of 5-H(p)ETE to 6-trans-LTB4 in a concentration-dependent manner. At low oxidant concentrations, there was a strong decrease in the yield of 6-trans-LTB4, whereas 5-HpETE did not change or increased. Additionally, the formation of 8-HpETE and 12-HpETE by 5-LOX rose slightly with increasing HOCl and HOBr. Comparable results were obtained with the MPO-H2O2-Cl– system when glucose oxidase and glucose were applied as a source of H2O2. This was necessary because of a strong impairment of 5-LOX activity by H2O2. In summary, MPO-derived oxidants showed a considerable impact on 5-LOX, impairing the epoxidation of 5-HpETE, whereas the hydroperoxidation of arachidonic acid was unaffected. Apparently, this was caused by an oxidative modification of critical amino acid residues of 5-LOX. Further work is necessary to assess the specific type and position of oxidation in the substrate-binding cavity of 5-LOX and to specify whether this interaction between 5-LOX and MPO-derived oxidants also takes place in stimulated neutrophils.

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Childs, Brian Richard. « Use of Personality Profile Assessments in the Construction Industry ». BYU ScholarsArchive, 2015. https://scholarsarchive.byu.edu/etd/5634.

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Various industries are using personality profile assessments as tools to help reduce employee turnover. Employee turnover is a continuing challenge in the construction industry that has negative effects on construction companies. Research found that it was difficult to know if companies in the construction industry are using personality profile assessments as a tool to help reduce employee turnover. After understanding that other industries were using personality profile assessments in their hiring, promoting, team building and leadership development to reduce turnover, it was desired to understand if the construction industry was doing the same. This research performed a survey among the top construction companies to understand if construction companies were using assessments, and if it had any effect on the turnover of those companies. The survey results provided information on the amount of companies using personality profile assessments, as well as additional insights and attitudes among these companies, whether they used assessments or not. The results of this survey and research have provided strong indicators that personality profile assessments are tools that will help construction companies reduce employee turnover.

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Robinson, Ailie. « An investigation into the effects of Plasmodium parasite infection on the human odour profile ». Thesis, London School of Hygiene and Tropical Medicine (University of London), 2017. http://researchonline.lshtm.ac.uk/4539377/.

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Some studies suggest that Plasmodium parasites can change the attractiveness of their vertebrate hosts to Anopheles vectors. This is suggestive of parasite manipulation, where the parasite alters a host phenotype to the parasite’s own fitness benefit. In this instance, Plasmodium parasites may benefit from increased transmission via a greater number of vector-host contacts. Despite evidence that variation in human host attractiveness to biting insects is manifest through differences in the human odour profile (volatile compounds produced by the skin), the association between odour profile and Plasmodium infection has never been studied in humans. The skin odour profile of individuals infected with Plasmodium in both experimental- and natural-infection settings was investigated using gas chromatography (GC). In the experimentally-infected (EI) cohort, adults were infected with Plasmodium falciparum and their odour profile sampled before, during and after infection. In the naturally-infected (NI) cohort, children aged 5-12 years with Plasmodium infections of varied parasite density, stage and species were sampled before and after treatment with antimalarials. Odour samples from both cohorts were further screened for infection-associated compounds using coupled GC-electroantennography (GC-EAG). In both EI and NI cohorts, changes in the production of several compounds in skin odour were found to be associated with the presence of Plasmodium parasites. Of these, production of the aldehydes heptanal, octanal and nonanal (C7-C9) was both increased in association with the presence of parasites in a density-dependent manner, and found to induce antennal response in Anopheles coluzzi. The production of C7-C9 and other infection-associated compounds via malaria-induced oxidative stress is a suggested mechanism. Malaria remains one of the most important diseases worldwide. As the global strategy for malaria control and elimination evolves, to combat both parasite and vector resistance to drugs and insecticides, the need for innovative tools intensifies. If malaria parasites can alter the human odour profile and attractiveness to mosquitoes, the repercussions would be far reaching: this would likely have a profound influence on the way that malaria spreads through populations, and malaria-specific volatile biomarkers could provide a basis for novel diagnostic tools.

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Fraser, Isla. « Establishing the relationship between environmental sources and the stable isotope profile of human tissues ». Thesis, Queen's University Belfast, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.479419.

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Aly, Amir. « Towards an Interactive Human-Robot Relationship : Developing a Customized Robot's Behaviour to Human's Profile ». Palaiseau, École nationale supérieure de techniques avancées, 2014. https://pastel.hal.science/tel-01128923.

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L'importance de considérer l'émotion dans l'interaction homme-robot comme base pour le comportement généré du robot, est la nature floue de l'émotion. Cela peut entraîner le robot à générer un comportement inapproprié au contexte de l'interaction en méconnaissant une émotion observée. Cela ouvre la porte vers une nouvelle méthodologie floue à base pour détecter l'émotion plus précisément. Cette méthodologie décide si l'émotion observée a besoin d'un nouveau comportement à synthétiser au cas où elle constitue une nouvelle classe non apprise auparavant, ou si elle peut être attribuée à un comportement existant dans la mémoire d'action du robot. D'autre part, l'effet à long terme de la personnalité sur le comportement verbal et non verbal de l’homme, le rend fiable pour être considéré comme un facteur déterminant pour le comportement multimodal synthétisé du robot. Par conséquent, l'adaptation du comportement généré verbal et non verbal du robot à la personnalité de l'homme comme étant introverti ou extraverti, pourrait bien améliorer l'attraction de l'homme au robot. Le processus de génération du comportement multimodal synchronisé du robot à travers la parole, les gestes, et les expressions faciales en fonction du profil de l'homme, subit un modèle cognitif de calcul. Ce modèle simule les fonctionnalités cognitives de l'homme, qui apprennent l'objectif et le mécanisme des actions multimodales effectuées par des hommes dans le milieu environnant. Par conséquent, lors d'une interaction, le robot devient capable de synthétiser par lui-même, un comportement multimodal basé sur le profil de l'homme, le contexte de l'interaction, et les expériences enregistrées dans sa mémoire d'action
Robots become more and more omnipresent in our life and society, and many challenges arise when we try to use them in a social context. This thesis focuses on how to generate an adapted robot’s behavior to human’s profile so as to enhance the human-robot relationship. This research addresses a wide range of complex problems varying from analyzing and understanding human’s emotion and personality to synthesizing a complete synchronized multimodal behavior that combines gestures, speech, and facial expressions. Our methodologies have been examined experimentally with NAO robot from Aldebaran Robotics and ALICE robot from Hanson Robotics. The first part of this thesis focuses on emotion analysis and discusses its evolutionary nature. The fuzzy nature of emotions imposes a big obstacle in front of defining precise membership criteria for each emotion class. Therefore, fuzzy logic looks appropriate for modeling these complex data sets, as it imitates human logic by using a descriptive and imprecise language in order to cope with fuzzy data. The variation of emotion expressivity through cultures and the difficulty of including many emotion categories inside one database, makes the need for an online recognition system of emotion as a critical issue. A new online fuzzy-based emotion recognition system through prosodic cues was developed in order to detect whether the expressed emotion confirms one of the previously learned emotion clusters, or it constitutes a new cluster (not learned before) that requires a new verbal and/or nonverbal action to be synthesized. On the other hand, the second part of this thesis focuses on personality traits, which play a major role in human social interaction. Different researches studied the long term effect of the extraversion-introversion personality trait on human’s generated multimodal behavior. This trait can, therefore, be used to characterize the combined verbal and nonverbal behavior of a human interacting with a robot so as to allow the robot to adapt its generated multimodal behavior to the interacting human’s personality. This behavior adaptation could follow either the similarity attraction principle (i. E. , individuals are more attracted by others who have similar personality traits) or the complementarity attraction principle (i. E. , individuals are more attracted by others whose personalities are complementary to their own personalities) according to the context of interaction. In this thesis, we examine the effects of the multimodality and unimodality of the generated behavior on interaction, in addition to the similarity attraction principle as it considers the effect of the initial interaction between human and robot on the developing relationship (e. G. , friendship), which makes it more appropriate for our interaction context. The detection of human’s personality trait as being introverted or extraverted is based on a psycholinguistic analysis of human’s speech, upon which the characteristics of the generated robot’s speech and gestures are defined. Last but not least, the third part of this thesis focuses on gesture synthesis. The generation of appropriate head-arm metaphoric gestures does not follow a specific linguistic analysis. It is mainly based on the prosodic cues of human’s speech, which correlate firmly with emotion and the dynamic characteristics of metaphoric gestures. The proposed system uses the Coupled Hidden Markov Models (CHMM) that contain two chains for modeling the characteristic curves of the segmented speech and gestures. When a speech-test signal is present to the trained CHMM, a corresponding set of adapted metaphoric gestures will be synthesized. An experimental study (in which the robot adapts the emotional content of its generated multimodal behavior to the context of interaction) is set for examining the emotional content of the generated robot’s metaphoric gestures by human’s feedback di- rectly. Besides, we examine the effects of both the generated facial expressions using the expressive face of ALICE robot, and the synthesized emotional speech using the text to speech toolkit (Mary-TTS) on enhancing the expressivity of the robot, in addition to comparing between the effects of the multimodal interaction and the interaction that employs less affective cues on human. Generally, the research on understanding human’s profile and generating an adapted robot’s behavior opens the door to other topics that need to be addressed in an elaborate way. These topics include, but not limited to: developing a computational cognitive architecture that can simulate the functionalities of the human brain areas that allow understanding and generating speech and physical actions appropriately to the context of interaction, which constitutes a future research scope for this thesis

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FANTINI, VALENTINA. « Functional analysis and transcriptome profile of meninges and skin fibroblasts from human aged donors ». Doctoral thesis, Università degli studi di Pavia, 2021. http://hdl.handle.net/11571/1446317.

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Garnett, Shaun. « Generating a proteomic profile of neurogenesis, through the use of human foetal neural stem cells ». Doctoral thesis, Faculty of Science, 2019. http://hdl.handle.net/11427/31143.

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Introduction Neurogenesis, the development of new neurons, starts soon after the formation of the neural tube and is largely completed by birth. Development of the brain after birth is mainly reliant on the formation of new connections between surviving neurons. However, adult neurogenesis does continue in the subgranular zone of the hippocampus from quiescent adult neural stem cells. Traditionally neural stem cells were cultured as neurospheres, a heterogeneous agglomeration of neural cells at various stages of differentiation. This heterogeneity prevented accurate quantitative analysis. In 2008 Sun et al produced the first non-immortalised human foetal neural stem (NS) cell line from nine week old human foetal cortex. These cells are cultured as monolayers, have a radial glia like appearance, self renew and form all three neural cell types, neurons, astrocytes and oligodendrocytes upon differentiation. More recently human foetal neuroepithelial like (NES) stem cells have been produced from five week old human foetal hind-brain, they resemble neuroepithelial cells, with characteristic rosettes, upon differentiation they appear to form a pure population of neurons. These homogeneous monolayer cultures enable quantitative proteomic analysis, to increase our understanding of early brain development Methods Three NES and two NS cell lines were available for analysis. They proliferate by stimulation from FGF and EGF, removal of these growth factors results in spontaneous differentiation. Proliferating NES and NS cells were compared using SILAC labelling. In addition, each cell line was differentiated for 12 days, 6 timepoints were taken and compared using label free quantitation. Results 4677 proteins were quantitated with 473 differentially expressed, revealing fundamental differences between NES and NS cells. NES cells are less differentiated, expressing SOX2 and LIN28, have active cell cycle processes, DNA elongation, histone modification and miRNA mediated gene silencing. Whereas NS cells are more developmentally defined, express multiple membrane proteins, have activated focal adhesion, thereby increasing their binding and interaction with their environment. NS metabolism is more oxidative, utilises lipid metabolism, the pentose phosphate pathway and produces creatine phosphate. Upon differentiation the cell cycle processes are downregulated and neurogenic and gliogenic processes increased. Conclusion This work represent a detailed in vitro characterisation of non immortalised human foetal neural stem cells, it describes the regulatory, metabolic and structural changes occurring within neural stem cells in early brain development. The information herein points towards de-differentiation potentially through LIN28-let7, as a means to produce more neurogenic neural stem cells in vitro thus aiding regenerative therapies, as well as provides a wealth of information for better understanding neurological developmental disorders.

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Frear, Susan W. « A Construct Validity Analysis of the Work Perceptions Profile Data ». Thesis, University of North Texas, 2015. https://digital.library.unt.edu/ark:/67531/metadc799499/.

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As work environments become more complex and demanding, organizations are becoming more interested in measuring the impact of their human resource development programs and initiatives. With this increased attention on data and measurement, human resource professionals have been encouraged to utilize data collection and data analysis techniques to make more objective and rationale human capital decisions and to verify business impact. As a result, the human resource profession has seen a significant increase in the use of surveys to measure anything from training effectiveness to the efficacy of recruitment procedures. The increase in the use of survey instruments requires that more focused attention is placed on the reliability and validity of data from any instrument used to make important human resource and business decisions. One instrument that is currently being used to measure career plateaus and job fit is the Work Perceptions Profile. The purpose of this research study was to conduct a construct validity analysis of the Work Perceptions Profile data and to determine the factor structure of data from its items. The data in this analysis supported a two-factor model structure with the first factor measuring Work Characteristics and a second factor measuring Performance. The results of this analysis will be helpful in exploring further how employees perceive their work place, their careers and their relationships with others within the organization.

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Dumont, Larry Joe. « Human cytomegalovirus reactivation following seasonal allergen exposure and switch to T-helper cell type 2 profile / ». Connect to full text at ProQuest Digital Dissertations. IP filtered, 2005.

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Thesis (Ph.D. in Clinical Sciences) -- University of Colorado at Denver and Health Sciences Center, 2005.
Typescript. Includes bibliographical references (leaves 140-156). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;

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Ding, Peihui, et 丁佩惠. « Expression profile, molecular regulation and immuno-inflammatory function of LPS-binding protein in human oral keratinocytes ». Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2012. http://hub.hku.hk/bib/B49617795.

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Lipopolysaccharide (LPS)-binding protein (LBP) functions as a crucial molecule in innate immune responses to bacterial challenge. Our recent study shows the expression of LBP in human gingiva and its significant association with periodontal condition. Porphyromonas gingivalis is a keystone periodontopathogen with its LPS as a major virulence factor strongly involved in periodontal pathogenesis. Recent study has discovered that P. gingivalis LPS displays a significant lipid A structural heterogeneity. The present study investigated i) the expression profile of LBP in human oral keratinocytes (HOKs) stimulated by P. gingivalis LPS with penta-acylated (LPS1690) and tetra- (LPS1435/1449) lipid A structures as well as E. coli LPS; ii) the involvement of toll-like receptors (TLRs) and downstream signaling mechanisms in LBP expression; and iii) the effects of LBP and its crosstalk with the two isoforms of P. gingivalis LPS on the expression of cytokines and human β-defensins (hBD-2) in HOKs. The expression of LBP mRNA and peptide was significantly up-regulated by P. gingivalis LPS1690 and E. coli LPS, while not by P. gingivalis LPS1435/1449. P. gingivalis LPS1690-induced LBP expression was through both TLR2 and TLR4, and the relevant down-stream signaling mechanisms were then further investigated. Western blot results showed that P. gingivalis LPS1690 activated the phosphorylation of IκBα, p65, p38 MAPK and SAPK/JNK, whereas E. coli LPS phosphorylated IκBα, p38 MAPK and SAPK/JNK. A nuclear translocation of NF-κB transcription factor was confirmed upon stimulation by both forms of LPS. Further blocking assay revealed that P. gingivalis LPS1690 induction of LBP was through NF-κB and p38 MPAK pathways, while E. coli LPS induction of LBP was mediated by NF-κB, p38 MPAK and JNK pathways. The effects of LBP and its crosstalk with P. gingivalis LPS1690 or LPS1435/1449 on the expression of cytokines and hBD-2 were further investigated. Interestingly, recombinant human LBP (rhLBP) per se could significantly up-regulate the expression of IL-6, IL-8 and TNF-α, while down-regulate hBD-2 expression. P. gingivalis LPS1690 or LPS1435/1449 modulated to different extents the rhLBP-induced cytokine expression. Notably, P. gingivalis LPS1690 significantly down-regulated rhLBP-induced IL-8 expression; whereas, P. gingivalis LPS1435/1449 down-regulated IL-8 expression more intensively (around 80% vs. 40% reduction). The key mediators of TLRs and their adaptors like CD180 and MD-1 were significantly down-regulated by rhLBP (fold changes: -2.44 and -9.62, respectively). Both CD180 and MD-1 mRNAs were up-regulated by P. gingivalis LPS1435/1449 (7.11 and 4.05 folds, respectively); while these two genes were reversely modulated by P. gingivalis LPS1690 (20.86 and -6.93 folds, respectively). The present study demonstrates that P. gingivalis LPS with a lipid A structural heterogeneity differentially modulates LBP expression in HOKs. P. gingivalis LPS1690 promotes LBP expression in HOKs through TLR2 and TLR4 as well as NF-κB and p38 MAPK pathways in a way different from E. coli LPS. rhLBP per se significantly up-regulates the expression of IL-6, IL-8 and TNF-α, while down-regulates hBD-2 expression. P. gingivalis LPS with different lipid A structures down-regulates to different extents the rhLBP-induced expression of cytokines in HOKs, likely through fine-tuning of the CD180-MD1 complex and the relevant TLRs.
published_or_final_version
Dentistry
Doctoral
Doctor of Philosophy

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Gravina, Lucia Concetta. « Imunohistochemical profile of the neuroblasts of the peripheral sympathetic nervous system and human neuroblastoma of childhood ». Doctoral thesis, Università di Catania, 2014. http://hdl.handle.net/10761/1537.

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ABSTRACT I focused on the analysis of a protein involved in the development of the peripheral nervous system: the Wilms tumor1 (WT1). My thesis is divided into two experimental phases: I) spatio-temporal distribution of WT1 during human embryonic development, II) expression and functional roles of WT1 during development of the peripheral sympathetic nervous system and gastrointestinal tract. The Wilms tumor (WT1) gene and its protein product are known to exhibit a dynamic expression profile during development and in the adult organism. Apart from a nuclear expression observed in the urogenital system, its precise localization in other developing human tissues is still largely unknown. Accordingly, the aim of this study was to investigate immunohistochemically the temporal and spatial distribution of WT1 in epithelial and mesenchymal developing human tissues from gestational weeks 7 24. For this purpose we used antibodies against the N-terminal of WT1. As might be expected, WT1 nuclear expression was observed in mesonephric/metanephric glomeruli, metanephric blastema, celomderived membranes (pleura, peritoneum, serosal surfaces) and sex cords. With regard to mesenchymal tissues, a similar nuclear staining was also obtained in the mesenchyme surrounding Müllerian and Wolffian ducts, as well as in the submesothelial mesenchymal cells of all celomatic-derived membranes. The most striking finding was the detection of strong WT1 cytoplasmic immunostaining in developing skeletal and cardiac muscle cells and endothelial cells. The tissue-specific expression of WT1, together with its different nuclear/cytoplasmic localization, both suggest that WT1 protein may have shuttling properties, acting as a protein with complex regulator activity in transcriptional/translation processes during human ontogenesis. The reported cytoplasmic expression of WT1 in human rhabdomyosarcomas and in many vascular tumors strongly suggests an oncofetal expression of this protein. Although not specific, WT1 cytoplasmic expression can be used as a marker of skeletal muscle and endothelial differentiation in an appropriate morphological context. Developmental expression of Wilms tumor gene (WT1) and protein is crucial for cell proliferation, apoptosis, differentiation and cytoskeletal architecture regulation. More recently, it has been suggested a potential role of WT1 in the development of neural tissue and in neurodegenerative disorders. We have investigated immunohistochemically the developmentally regulated expression and distribution of WT1 in human fetal (from the 8th to the 28th week gestational age) peripheral sympathetic nervous system (PSNS) and gastro-enteric nervous system (GENS). Interestingly WT1 expression was restricted to the cytoplasm of sympathetic neuroblasts, while it progressively disappeared with advancing morphologic differentiation of these cells along both ganglionic and chromaffin cell lineages. In adult tissues, both ganglion and chromaffin cells lacked any WT1 expression. These findings show that WT1 is a reliable marker of human sympathetic neuroblasts, which can be used routinely in formalin-fixed, paraffin-embedded tissues. The progressive loss of WT1 in both ganglion and chromaffin cells, suggests its potential repressor role of differentiation in a precise temporal window during human PSNS and GENS development.

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MERAVIGLIA, VIVIANA. « Human mesenchymal stromal cells : how the tissue of origin influences plastic properties and microRNA expression profile ». Doctoral thesis, Università degli Studi di Milano-Bicocca, 2014. http://hdl.handle.net/10281/51144.

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MicroRNAs are key modulators at molecular level in different biological processes, including determination of cell fate and differentiation. Herein, microRNA expression profiling experiments were performed on syngeneic cardiac and bone marrow mesenchymal stromal cells cultured in standard growth medium and then in vitro exposed to adipogenic, osteogenic, cardiomyogenic and endothelial differentiation media. Analysis identified a tissue-specific microRNA signature composed by 16 microRNAs that univocally discriminated cell type of origin and that were completely unaffected by in vitro differentiation media: 4 microRNAs were over-expressed in cardiac stromal cells, and 12 were overexpressed or present only in bone marrow stromal cells. Further, results revealed microRNA subsets specifically modulated by each differentiation medium, independently from the cell type of origin, and a subset of 7 microRNAs that were downregulated by all media with respect to growth medium. Finally, we identified 16 microRNAs that were differentially modulated by the media when comparing the two tissues of origin. The existence of a tissue-specific microRNA signature surviving to any differentiation stimuli, strongly support the role if microRNAs determining cell identity related to tissue origin. Moreover, we identified microRNA subsets modulated by different culture conditions irrespective of tissue origin, pointing out their importance during differentiation processes.

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Llewellyn, Bonny C. « A Profile and Analysis of Repeat Bankruptcy Petitioners in the District of Utah 1984-2004 ». DigitalCommons@USU, 2005. https://digitalcommons.usu.edu/etd/2849.

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The purpose of this study was to describe the incidence of repeat filers in Utah and estimate the extent that repeat filers may be abusing the bankruptcy system. This study sought to develop a profile of repeat filers . Demographic and financial variables were examined to detem1ine their association with abuser/nonabuser status. In this study, abuse of bankruptcy was characterized only by the timing and number of filings over 20 years. Debtors with three filings in a 2-year period or less and debtors with four or more total filings were classified as abusers. Nonabusers were defined as debtors who filed only once as well as debtors who had two or three scattered filings over the 20-year period. About I I% of the total sample appeared to be abusing the bankruptcy system by filing repeatedly. The majority (76.2%) of the I 997 cases filed by abusers were dismissed while only 23.8% received a discharge of their unsecured debts. Only five (2.9%) of the I 71 abusers who filed chapter I 3 in I 997 completed their payment plan and received a discharge of their debts. The logistic regression model found chapter (7 versus 13), filing status, unsecured debt, and monthly income to be the most significant variables in estimating abuse. Males and females filing alone were nearly 50% less likely than joint filers to be abusers. Chapter 13 debtors are nearly five times as likely to be abusers when compared to chapter 7 debtors. Filers who had unsecured debt levels above the median were less likely to be abusers, and filers who had incomes above the median were almost twice as likely to be abusers. Realistic repayment plans that pay careful attention to construction of budgets and a financial counselor to work with debtors who miss payments is one approach to combating abuse by repeat filing. Judges may need to discipline attorneys who file cases repeatedly. Perhaps a new Code is not what we need to combat abusers; instead, closer monitoring of cases by trustees, more responsible attorneys, and more responsible lending are needed.

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Tuccitto, Alessandra. « pH regulatory molecules in the tumour microenvironment : modulators of aggressiveness and immune profile of human hepatocellular carcinoma ». Thesis, Open University, 2018. http://oro.open.ac.uk/55985/.

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Background: Hepatocellular carcinoma (HCC) arises in an inflammatory, hypoxic/acidic microenvironment that favours tumour progression and fosters immunosuppression. Tumour cells survive this hostile environment by over-expressing pH regulatory molecules such as carbonic anhydrase (CA) IX, XII and V-ATPase complex, but the relevance of these molecules in HCC is poorly defined. Aim: The aim of this study was to dissect the relationships between pH regulatory molecules and the aggressive behaviour of malignant hepatocytes, and to evaluate how pH regulatory molecules influence the immune microenvironment of HCC. Methods: HCC, non-tumour and normal liver tissue samples were analysed by qRT-PCR for the expression of genes encoding the pH regulatory molecules (CAIX, CAXII and V-ATPase), of genes associated to epithelial-to-mesenchymal-transition (EMT) (TWIST, CDH1, VIM) and those encoding for HCC stem cell-associated markers (CD13, CD24, CD44, CD90, EpCAM, CD133, KRT19, OCT4, NANOG and SOX2). Selected HCC, non-tumour and normal liver tissue samples were evaluated by immunohistochemistry (IHC) to detect the presence and localization of CAIX, CAXII and VATPase and to assess the distribution of macrophages and T cells. Confocal microscopy and flow cytometry were implemented to assess the co-expression of selected markers. HCC cell lines, characterised for the expression of pH regulators, were tested for the sensitivity to the CAIX, CAXII, and V-ATPase specific inhibitors. The effects of V-ATPase specific drug were also studied ex vivo in primary human HCC tumour explants by qRT-PCR and by flow cytometry in HCC single cell suspensions obtained by the enzymatic digestion of HCC specimens. Results: Our mRNA analysis showed that the expression of CA9 was significantly correlated with the expression of the hypoxia-inducible factor 1α-related gene (HIF1A) and of the stem cell-associated markers CD24, CD133, EpCAM and KRT19. Moreover, mRNA for CA9 and for the different CA12 isoforms were associated with tumour grading, thus indicating their possible role in tumour malignancy. Applying a machine learning tool known as the ‘Adaptive Index Model’ the combined expression of different CA12 isoforms, CD209 and CDH1 defined a ‘signature’ classifying HCC patients in groups at different risk of recurrence, thus indicating a link between pH regulators, myeloid and EMT markers likely influencing HCC prognosis. IHC analysis indicated that HCC displays a complex expression pattern for the pH regulatory proteins. Both CAIX and CAXII were detected in transformed, but not in normal hepatocytes. CAIX protein had a focal distribution in the tumour, thus supporting its possible association with hypoxic and the most aggressive tumour area. Conversely, CAXII was homogeneously expressed by all tumour hepatocytes, but mainly retained in the endoplasmic reticulum (ER). The majority of HCC expressed V-ATPase which, importantly, was also present in immune infiltrating cells. This expression pattern qualified the CAIX, CAXII and V-ATPase as possible targetable molecules. Our in vitro data indicated that blockage of their enzymatic activities by specific drugs affected the viability of HCC cell lines in a dose dependent fashion, although with the CAXII specific inhibitor showing low efficacy, likely related to the preferential ER localization of CAXII molecules inside the HCC cells. Ex vivo experiments with HCC tissue explants and HCC cellular suspensions showed that inhibition of VATPase modulated the epithelial/mesenchymal features of HCC cells and the levels of pro- and anti-tumour cytokines expressed by M2 macrophages and T cells infiltrating HCC. Conclusions: Herein, our data demonstrated that the pH regulatory molecules, CAIX, CAXII and V-ATPase are over expressed in the HCC microenvironment and interfering with their pathways exerted anti-tumour activities, although these data also lead to the conclusion that more effective CAXII specific drugs should be designed. The results of this thesis also suggest that pH regulatory molecules might have a role in HCC aggressiveness and prognosis. Importantly, one of these pH regulators, namely V-ATPase complex, influences the mesenchymal features of tumour cells and the immunosuppressive tumour microenvironment (TME). Interfering with tumour metabolism is an emerging strategy for treating cancers that are resistant to standard therapies. Thus, targeting the unique crosstalk between tumour cells and the microenvironment, played by the pH regulatory molecules, can be considered as a new option for HCC treatment and the blockage of the V-ATPase complex might represent a multi-task strategy for the treatment of HCC patients.

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Urquhart, Paula, Susan M. Parkin et Anna Nicolaou. « Profile of eicosanoids produced by human saphenous vein endothelial cells and the effect of dietary fatty acids ». Harcourt, 2009. http://hdl.handle.net/10454/4038.

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no
Human saphenous vein endothelial cells (HSVECs) derived from primary cultures of adult human veins constitute an excellent in vitro model for studying human endothelial metabolism. In this study we report the14C-labelled prostanoid profile of HSVECs under resting and stimulated conditions and the effect of the n-3 polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid on them. Results indicate that HSVECs while under resting conditions produce mainly prostaglandin F2 ¿(PGF2 ¿). After stimulation with calcium ionophore A23187, the cells were found to synthesise PGI2, PGE2and PGF2¿as major products and thromboxane B2and PGD2as minor products. Production of14C-labelled hydroxyeicosatetraenoic acids was not detected. Eicosapentaenoic acid was found to inhibit basal and stimulated prostanoid production whereas docosahexaenoic acid inhibited basal but strongly increased stimulated prostanoid production. These results may offer the basis for further studies aiming to investigate targets for pharmacological intervention in inflammatory conditions.

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Fiorelli, S. « BIOCHEMICAL AND FUNCTIONAL PROFILE OF SPONTANEOUSLY DIFFERENTIATED HUMAN MONOCYTE-DERIVED MACROPHAGES IN PATIENTS WITH ISCHEMIC HEART DISEASE ». Doctoral thesis, Università degli Studi di Milano, 2018. http://hdl.handle.net/2434/544434.

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Background: Atherosclerosis is a chronic inflammatory disease in which oxidative stress and macrophages play fundamental roles. Macrophages are heterogeneous cells in term of morphology and function and the prevalence of a specific morpho-phenotype may influence the progression or the regression of the plaque. Human coronary plaque macrophages are not easily obtainable and monocyte-derived macrophages (MDMs) are widely accepted as a good surrogate. Our previous study has reported that healthy subjects show two dominant MDM morphotypes, spindle and round, present in the same percentage. Aim: The aims of the study were to assess the biochemical and functional profile of MDMs obtained from CAD patients and to investigate the relationship between these characteristics and plaque features detected in vivo. Material and methods: ninety CAD patients and twenty-five healthy volunteers were enrolled. MDMs were obtained by culturing monocytes for 7 days in medium supplemented with 10% autologous serum. Transglutaminase2 (TG2), tissue factor (TF), nuclear factor erythroid 2–related factor 2 (Nrf2) and heme-oxygenase (HO-1) were determined by western blotting and immunofluorescence. The uptake of apoptotic Jurkat T cells was detected by flow cytometry and thrombin formation was assessed by thrombinoscope. Fatty acids composition and oxidative stress status, determined as the ratio between the reduced and oxidized forms of glutathione (GSH and GSSG, respectively) were analyzed by liquid-chromatography tandem mass spectrometry (LC-MS/MS). Plaque features were detected in vivo by optical coherence tomography (OCT). Results: MDMs obtained from CAD patients were characterized by the predominance of round cells. These cells exhibited a lower efferocytic capacity and higher capacity to generate thrombin in respect to those of healthy subjects, reflecting the low expression of TG2 and the enhance of TF levels. Moreover, MDMs of CAD patients showed a higher oxidative stress status, evidenced by the lower GSH/GSSG ratio, in respect to those of healthy subjects. Finally, we found a positive correlation between oxidative stress status and membrane fluidity (MFI) as evidenced by the ratio of C18:1 (oleic acid)/C18:2 (linoleic acid), as well as MFI and capacity to engulf apoptotic cells. Regarding OCT analysis, patients with a higher round MDMs prevalence exhibited more frequently a lipid rich plaque, a TCFA (thin cap fibroatheroma), a greater intra-plaque macrophage content, and a ruptured plaque, characteristics of vulnerable plaque. Furthermore, patients with high TF levels more frequently presented a intra-plaque macrophages accumulation, a ruptured plaque and a presence of thrombus. In addition, vulnerable plaque characteristics were associated with high HO-1 levels. Conclusions: MDMs of CAD patients present a pro-inflammatory and high thrombogenic profile in respect to those of healthy volunteers. The high oxidative stress present in these cells influences the membrane fluidity that in turn influences the efferocytic capability. This specific cellular profile detected in in vitro obtained MDMs is associated with the detection of high-risk and rupture-prone coronary plaques in vivo, at OCT investigation. Different MDM phenotypes might be novel diagnostic and therapeutic target able to counteract the progression of atherosclerosis.

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MEREGALLI, CHIARA. « STUDY OF MULTIPOTENT RENAL PKHHIGH STEM-LIKE CELLS, ISOLATED FROM HUMAN NEPHROSPHERES : REGENERATIVE ABILITIES AND TRANSCRIPTOMIC PROFILE ». Doctoral thesis, Università degli Studi di Milano-Bicocca, 2018. http://hdl.handle.net/10281/199041.

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Il meccanismo alla base della riparazione di un danno a cellule renali è ancora oggi oggetto di dibattito e potrebbe coinvolgere sia cellule terminalmente differenziate che l'esistenza di cellule staminali multipotenti quiescenti. Sfruttando il saggio di formazione di sfere e la tecnica del FACS sorting, il nostro gruppo ha isolato una popolazione di cellule marcate col clorante PKH26 che avessero caratteristiche di cellule staminali renali adulte (PKHhigh). In precedenza abbiamo valutato la capacità di queste cellule di differenziare in vitro in lineage epiteliale, podocitico ed endoteliale. Abbiamo anche dimostrato che la popolazione PKHhigh è eterogenea nella sua composizione e all'interno delle nefrosfere le cellule con capacità simil-staminali sono PKHhigh/CD133+/CD24- (RSC). Abbiamo recentemente pubblicato che le nostre cellule delle nefrosfere, che comprendono cellule PKHhigh e la loro progenie PKHlow/neg, sono in grado di ripopolare scaffold renali umani decellularizzati. Con questa ricerca, ora abbiamo come scopo: i) di dimostrare le capacità rigenerative delle cellule staminali PKHhigh, anche in assenza della loro progenie PKHlow/neg. ii) di trovare la signature molecolare delle RSC che, tra PKHhigh, sono le cellule con capacità staminali più ampie. Per raggiungere questi obiettivi, abbiamo messo in coltura cellule PKHhigh su scaffold acellulari per 30 giorni e le cellule delle strutture ripopolate sono state caratterizzate mediante immunofluorescenza sequenziale, utilizzando specifici marcatori di differenziamento. Alcune strutture hanno indicato un differenziamento terminale in lineage epiteliale tubulare prossimale o distale o in quello endoteliale. Solo poche strutture mostravano invece la coespressione di alcuni o di tutti i marcatori testati, suggerendo un fenotipo ancora immaturo. Per la comprensione della signature molecolare delle RSC, è stata eseguita l'analisi trascrittomica delle RSC stesse, della loro progenie PKHlow/neg e di colture primarie terminalmente differenziate e sono stati evidenziati geni differenzialmente espressi (DEG). L’analisi bioinfomatica mediante Gene Set Enrichment Analysis ha suggerito uno stato immaturo delle nostre RSC, ma comunque diverso da cellule staminali embrionali. Infine, incrociando le liste di DEG sono stati ottenuti potenziali marcatori e alcuni di essi sono stati selezionati e validati. In conclusione, abbiamo evidenziato le capacità differenziative in senso epiteliale sia prossimale che distale ed endoteliale delle cellule PKHhigh. Inoltre, abbiamo dimostrato che le cellule PKHhigh, completamente prive di qualsiasi marcatore endoteliale, sono in grado di dare origine a strutture simil-endoteliali. La coespressione occasionale di marcatori epiteliali ed endoteliali in strutture ripopolate potrebbe indicare uno stato precoce di transizione verso un differenziamento terminale. Il possibile ruolo della progenie PKHlow / neg nel velocizzare il differenziamento terminale delle cellule PKHhigh dovrà essere chiarito. Infine, la signature delle RSC ottenuta potrà aprire la possibilità di isolamento diretto di cellule staminali renali adulte da tessuto renale normale.
The mechanism underlying the recovery of renal cell injury is still a matter of debate that concerns the involvement of fully differentiated cells, or the existence of quiescent scattered multipotent stem cells. By sphere forming assay and sorting, our group isolated a population of PKH26 most fluorescent cells with characteristics of adult renal stem-like cells (PKHhigh cells). We previously assessed the ability of PKHhigh cells to differentiate in vitro along epithelial, podocytic and endothelial lineages. We also demonstrated that PKHhigh population is heterogeneous in composition and within nephrospheres the cells with stem capacities are PKHhigh/CD133+/CD24- (RSC). We recently published that our nephrosphere cells, comprising PKHhigh cells and their PKHlow/neg progeny, are able to repopulate human decellularized renal scaffolds. With this research, we now aim: I) to prove the regenerative capabilities of PKHhigh stem-like cells, even in absence of their PKHlow/neg progeny. II) to find the molecular signature of RSC that among PKHhigh cells are those with the wider stem capacities. To reach these aims, we cultured isolated PKHhigh cells on acellular scaffolds for 30 days and the cells of the repopulated structures were characterized by sequential immunofluorescence using specific markers of differentiation. Some structures indicated a specific lineage differentiation into proximal and distal tubular epithelium and endothelium. Only few structures coexpressed some or all markers tested, indicating still immature phenotypes. For the disclosure of RSC molecular signature, transcriptomic analysis of RSC, of their PKHlow/neg progeny and of terminally differentiated primary cell cultures (PCC) was performed and differentially expressed genes (DEG) were evidenced. Bioinformatic Gene Set Enrichment Analysis suggested a renal immature status of our RSC, but different from embryonic stem cells. Crossing DEG lists potential markers were obtained and some of them were selected and validated. In conclusion, we highlighted the proximal and distal tubular epithelial and endothelial differentiative and regenerative abilities of PKHhigh cells. Moreover, we showed that PKHhigh cells, completely lacking any endothelial marker, were able to give rise to endothelial-like structures. The occasional coexpression of epithelial and endothelial markers in repopulated structures may indicate a transitional early status toward cell differentiation. The eventual role of the PKHlow/neg progeny of PKHhigh cells in speeding up the complete PKHhigh differentiation will be clarified. Finally, the obtained RSC signature would open the possibility for the direct isolation of adult renal stem-like cells from normal kidney tissue.

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Wollin, Judy A. « Assessing disability : the reliability and validity of the multiple sclerosis disability profile ». Thesis, Queensland University of Technology, 1998. https://eprints.qut.edu.au/36737/1/36737_Digitised%20Thesis.pdf.

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The assessment of people with disabilities is complex. The priorities adopted and issues addressed vary. The Multiple Sclerosis Disability Profile was developed and is currently used to assess people with MS in Australia. However, little is known about its psychometric properties and people with MS have not had a previous opportunity to comment on its relevance to them. Therefore the reliability and validity of the Disability Profile was assessed using widely used assessment methods and the views of people with MS. Standard methods for assessing the characteristics of assessment scales were used and one hundred and three people with MS were asked to nominate important issues that need discussing when assessing the person with MS. Support people and health professionals provided additional views. The research was conducted from a theoretical perspective that considers the relationship between disability and citizenship. The internal consistency of the Disability Profile was found to be excellent (Cronbach's Alpha= 0.9656). Furthermore it was found that the inter-rater reliability of the Multiple Sclerosis Disability Profile can be improved with clearer operational definitions, increased training for health professionals in the use of the instrument and reduced number of responses per item. Moreover the assessment of disability needs to be improved to achieve more sensitivity and to incorporate the handicap score devised in the course of this study. These amendments will enable improved discrimination when assessing handicap. However further research is required to assess the reliability and validity of the amended instrument. As the Disability Profile is a starting point for assessment, therapy and support for the person with MS, the revisions proposed here offer health professionals a holistic assessment instrument with acceptable reliability and viability.

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Mayta-Tristan, Percy, Elías Reneé Pereyra, Juan José Montenegro-Idrogo, Christian R. Mejia, Berrospi Fiorella Inga et Holguín Edward Mezones. « Profile and professional expectations of medical students from 11 Latin American countries : the Red-LIRHUS project ». Biomed Central Ltd, 2017. http://hdl.handle.net/10757/622009.

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Background Latin America is undergoing a human resource crisis in health care in terms of labor shortage, misdistribution and poor orientation to primary care. Workforce data are needed to inform the planning of long-term strategies to address this problem. This study aimed to evaluate the academic and motivational profile, as well as the professional expectations, of Latin American medical students. Results We conducted an observational, cross-sectional, multi-country study evaluating medical students from 11 Spanish-speaking countries in 2011–2012. Motivations to study medicine, migration intentions, intent to enter postgraduate programs, and perceptions regarding primary care were evaluated via a self-administered questionnaire. Outcomes were measured with pilot-tested questions and previously validated scales. A total of 11,072 valid surveys from 63 medical schools were gathered and analyzed. Conclusions This study describes the profile and expectations of the future workforce being trained in Latin America. The obtained information will be useful for governments and universities in planning strategies to improve their current state of affairs regarding human resources for health care professions.

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Archontoulis, Fiona L. « The implementation of a workload allocation system in pursuit of employee health outcomes ». Thesis, Griffith University, 2020. http://hdl.handle.net/10072/395540.

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The impact of increasing global competitiveness and the ensuing decisions taken by organisations to adjust, often have negative impacts on the health and wellbeing of employees. Increasing workloads, time and resourcing constraints are contributing factors to changing dynamics and psychosocial characteristics in many workplaces. There is ample evidence about the cost to organisations, society and individuals when financial performance objectives are prioritised over employee health and well-being. Human resource management (HRM) studies commonly address organisational performance outcomes overshadowing the importance of workplace health policies and procedures. Yet, extant literature reviewed from a variety of disciplines suggest effective HRM requires consideration of equity, fairness, job design, work stress and role expectations are important variables for employee health and well-being. Significantly for this study, the psychological health of employees, like physical safety, has gained increasing focus with legislation protecting employees against psychosocial hazards. Excessive workloads, work intensification, reduced recovery time and technology based systems supporting employees to work from home are all hazards associated with poor employee health and blur the boundaries between work and home. The Australian higher education sector has not been spared the impacts of global competitiveness. Over the last four decades, the notion of a collegial, autonomous academic profession has gradually transitioned to an environment more aligned with corporatisation and managerialism. As a result, the sector has morphed into a highly competitive international market for higher education resulting in poor health and well-being outcomes for academic employees. There is much debate in the literature about the health of academics, increasing workloads and associated effectiveness of workload allocation systems utilised in many universities. Yet there is minimal examination about how the systems could be positively utilised to support health and well-being outcomes for academics. Equally important, yet also not adequately understood is the role of frontline managers in higher education, their implementation of HR practices, the challenges they face in that, and the support they are afforded by senior management. This thesis addresses the issues highlighted above with an integrated HRM and workplace health perspective applied to a large Australian university (The University). A work profile system (WPS) is the HR practice of allocating work to academic employees at The University and is the unit of analysis for this study. An inductive, multi-level case study approach was adopted for this thesis where qualitative data was collected from 52 semi-structured interviews. A review of documentation (both publicly available and confidential to the organisation) contributed to the data collection process. Senior management/HR personnel representatives comprised Study A (Designers) so as to gain a retrospective understanding of the factors considered by The University in its initial decision to introduce a WPS. Secondly, frontline managers (FLMs) comprised Study B (Implementers) providing insights into the implementation challenges and opportunities of the system. Thirdly, full time tenured academic employees (Study C - Experiencers) provided first-hand accounts of working within the stipulations of the WPS. The research design enabled cross study analysis of data, and an investigation of the progression of policy development to implementation of a key HR practice at The University. This is an important contribution to knowledge providing a more holistic understanding of the why (the intention behind the WPS), the how (implementation practicalities) and the what (the actual experiences of the WPS). The thesis also finds internal contextual factors such as The University’s psychosocial climate; unique job design of an academic’s role; unique role of FLMs in academia and the power and authority delegated to them, all have significant influence on how the WPS can support better health outcomes of academic employees. Two theories were used to guide the study. The process model of strategic HRM (Wright and Nishii, 2007) provided a framework for data collection. The Psychosocial Safety Climate (PSC) theoretical model (Dollard & Bailey, 2014) incorporating the Job Demands-Resources (JD-R) model (Bakker & Demerouti, 2007) was the overarching theoretical guide for the study. The PSC theory also provided a foundation for the development of the sub-research questions for each of the three studies. Application of the PSC theory highlighted how management driven HR policies, practices and procedures for the protection of worker health can determine employees’ perceptions of an organisation’s psychosocial safety climate. Additionally, how the PSC can impact the level of receptivity of HR practices by employees. However the PSC theory failed to explain a number of findings including why the PSC at The University can be misinterpreted when there is some evidence health and well-being concerns were behind The University’s decision to introduce the WPS. The PSC theory was also unable to explain how external drivers had contributed to the decision to introduce the WPS. The findings did however highlight the spectrum of views among Designers in Study A suggesting a lack of consensus about the WPS. The JD-R component of the PSC theory was also unable to explain a disconnect between the views of some Designers that the WPS was a useful resource for academics to manage their workloads and the views of most Study B and Study C participants who perceived the WPS as a job demand. As would be expected from the relevant theories and other research, many findings from this study are consistent and support the extant literature. However, due to the PSC theory being unable to fully answer the research question, an extension to the PSC theoretical model is proposed as the contribution to this study. If the principles of the HRM systems strength model, developed by Bowen and Ostroff (2004), are applied as an antecedent to the PSC theory, with more focus on HRM system design, implementation and feedback processes it is proposed both theories combined can provide the answer to the RQ: How can a WPS support health and well-being outcomes of academics? People acting in the role of FLM in academia perform the role, often on a short term basis with the intention of reverting back to a frontline academic position. This arrangement often leads to a perceived lack of relevance of the WPS for operational needs and consequently a lack of consistency in implementation practices. While requiring a high level of management skills in implementing the WPS and managing their teams, FLMs are often not skilled to do so or do not have management inclinations. This was reflected in the data obtained from academic employees where they perceived their FLMs lacked authority in their role and therefore were not able to formally make adjustments to better support the health and well-being of their teams. This study also identified that FLMs did not always have the required support of senior management to formally adjust resourcing where necessary. This resulted in frustrations and many time consuming work arounds often impacting negatively on their own health and well-being. The general view expressed by participants of Study C was the WPS does not accurately reflect all the responsibilities of their role. More so however, reduced autonomy over their work, their FLMs unable to make necessary adjustments to workloads and perceptions senior management were not listening suggested a low psychosocial safety climate. In summary, this thesis identifies the need to adopt the principles of the HRM systems strength approach when introducing and implementing new HR practices. Consensus and consistency in messaging amongst HR/senior management; ongoing support for implementers of the practice; and a perceived distinctive relevance of the HR practice by employees are factors required to achieve intended outcomes. It was also identified improved management driven opportunities for academics to provide feedback about workloads, their workplace health and input for process improvements would contribute to a positive psychosocial safety climate at The University.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
Dept Empl Rel & Human Resource
Griffith Business School
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Mannem, Chandana. « IN-QUEST OF BIOMARKERS FOR ALZHEIMER’S DISEASE AND PHARMACOKINETIC PROFILE OF ANTICANCER AGENTS USING LC-MS IN HUMAN PLASMA ». Cleveland State University / OhioLINK, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=csu1579470084334844.

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Shimura, Kazuya. « Broad antiretroviral activity and resistance profile of the novel human immunodeficiency virus integrase inhibitor elvitegravir (JTK-303/GS-9137) ». Kyoto University, 2008. http://hdl.handle.net/2433/135828.

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Kepenekci, Burcu. « Human Activity Recognition By Gait Analysis ». Phd thesis, METU, 2011. http://etd.lib.metu.edu.tr/upload/12613089/index.pdf.

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This thesis analyzes the human action recognition problem. Human actions are modeled as a time evolving temporal texture. Gabor filters, which are proved to be a robust 2D texture representation tool by detecting spatial points with high variation, is extended to 3D domain to capture motion texture features. A well known filtering algorithm and a recent unsupervised clustering algorithm, the Genetic Chromodynamics, are combined to select salient spatio-temporal features of the temporal texture and to segment the activity sequence into temporal texture primitives. Each activity sequence is represented as a composition of temporal texture primitives with its salient spatio-temporal features, which are also the symbols of our codebook. To overcome temporal variation between different performances of the same action, a Profile Hidden Markov Model is applied with Viterbi Path Counting (ensemble training). Not only parameters and structure but also codebook is learned during training.

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Guéron, David. « Modélisation d'activités et agrégation de profils de vol ». Thesis, Aix-Marseille 3, 2011. http://www.theses.fr/2011AIX30044.

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L'agrégation d'activités pour l'identification de catégories de comportements est un enjeu majeur de tous les systèmes socio-techniques complexes actuels. La question clé consiste à réaliser une synthèse de façons de faire (ou praxies) intégrant la variabilité des opérateurs humains impliqués. Dans un cadre aéronautique, l'agrégation d'activités de pilotage vise à accélérer la détermination de procédures améliorant la sécurité des vols et l'efficacité des missions ; elle repose sur les données objectives des paramètres enregistrés des phases de vol significatives et se structure grâce à une interprétation experte. Un modèle d’Agrégation Supervisée : - décomposition, - maïeutique, - reconstruction, est ainsi établi dans cette thèse. Le cœur en est la 2e étape qui généralise et enrichit le concept de « moyenne » classique des approches probabilistes : une base d'apprentissage, constituée d'activités déterminées et caractérisées par l'interprétation experte, est utilisée pour identifier les motifs significatifs de paramètres enregistrés, c'est à dire les praxies qui agrègent donc les éléments essentiels des activités. Ceux-ci sont choisis au sein d'un ensemble de motifs paramétrables génériques, dont les divers seuils sont ajustés de manière incrémentale. Les motifs sont alors évalués selon les deux critères intrinsèques de cohérence et de pertinence de leurs seuils, ainsi que le critère extrinsèque de la conformité des résultats obtenus par leur utilisation aux vols de la base d'apprentissage. Peuvent à ce niveau se faire jour des groupements parmi les éléments de la base d'apprentissage, selon les motifs rendant compte des activités particulières. L'expertise doit également être généralisable pour permettre l'étude de plusieurs points-clé dans cette étape maïeutique.Ce modèle générique définit une activité comme une structure formelle de praxies, et ouvre la voie à un enrichissement de la 3e étape intégrant la multiplicité des rôles des opérateurs
Aggregating activities in order to identify categories of behaviour is a major topic of actual complex socio-technical systems. The key issue lies in incorporating the variability of implied human operators in the synthesis of ways of doing (or praxis). Aggregation of piloting activities is directed to allow a faster and more secure determination of procedures enhancing flight security and mission efficiency; it is based on the objective data of flight parameters recorded during significant flight phases, and is carried under thorough expert interpretation.A Supervised Aggregation model, consisting in the 3 steps of 1) decomposition, 2) maieutics, and 3) reconstruction, is thus devised in the present PhD. At the heart of this aggregation process, the 2nd maieutic step generalizes and enriches the usual concept of ''mean'', deeply related to probabilistic approaches: a set of activities analyzed and characterized by the expert, the learning basis, is related to significant patterns in the lot of recorded flight parameter values, in other words the praxis resulting of the aggregation of the activities. The patterns are selected from a collection of customizable generic patterns, whose thresholds are incrementally adjusted using the learning basis. The obtained patterns are then assessed according to the three criteria of 1) coherence and 2) likelihood of the thresholds, as well as the 3) conformity of these patterns used on the learning basis. At this stage, groups among the studied behaviours might emerge, gathering those for which an activity would be depicted by similar patterns. Expert-knowledge must be generalized in order to perform the joint analysis of several key points in this maieutic step.This generic model defines an activity as a formal structure of praxis, paving the way towards the further developments of the process, through the enrichment of the 3rd step, incorporating the multiplicity of operating roles

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Kincer, Caroline D. « A Paleodemographic Analysis of a Sample of Commingled Human Skeletal Remains at Ohio University ». Ohio University Honors Tutorial College / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=ouhonors1524843441411637.

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Sugden, Heather. « High resolution palynological, multiple profile and radiocarbon dating studies of early human impacts and environmental change in the Inner Hebrides, Scotland ». Thesis, University of Sheffield, 1999. http://etheses.whiterose.ac.uk/10274/.

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The Inner Hebrides comprise a diverse range of environments and vegetation, andarchaeological evidence suggests that people were interacting with these from early Holocene times. There are relatively few detailed palynological investigations from the islands for the early Holocene and not all the published data include quantification of microscopic charcoal which may assist in the interpretation of human impacts. Radiocarbon dates are also lacking from a number of the published profiles so that inter-site comparisons and comparisons with the archaeological record are difficult. Some pollen profiles from the Inner Hebrides contain possible indications of human impact in the first half of the Holocene (Lowe and Walker, 1986a; Andrews et al., 1987; Herons and Edwards, 1990; Edwards and Berridge, 1994). These profiles lack detail however, and it was clear that a multiple profile approach would provide a clearer picture of vegetation change and allow more confident interpretations of the pollen data. The coring of several sites would assist in defining the spatial differences in early Holocene vegetation within the islands and differences in the scale of human impacts which may reflect different types of interference. Multiple profiles were obtained from Loch a'Bhogaidh (Islay), A'Chrannag bog and Livingstone's Cave bog (Ulva) and Kinloch (Rum), all of which are close to areas of known Mesolithic occupation. A single core was obtained from Loch an t'Suidhe at Bunessan, south west Mull, from where there is currently no archaeological record for the Mesolithic. All cores were analysed for pollen and microscopic charcoal and AMS dates were obtained for all profiles. The results provide evidence for changes in vegetation due to climatic impacts and inferred Mesolithic activity. The possible effects of human influence vary from temporary woodland reductions to the creation of heathland and cereal cultivation. The use of multiple profiles is validated in that it provides an indication of spatial variation in inferred land use patterns over short timescales. The results are compared with previously published studies and the factors influencing the early Holocene spread of arboreal taxa, and the elm decline, are re-evaluated.

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Goeiman, Hilary. « Developing a human resource profile for the nutrition workforce in the public health sector in the Western Cape province, South Africa ». Thesis, Link to the online version, 2008. http://hdl.handle.net/10019/1750.

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Ogunrombi, Modupe Olufunmilayo. « The effect of grapefruit juice, a P-glycoprotein inhibitor, on organic acid and conjugates urinary profile in healthy human subjects / M.O. Ogunrombi ». Thesis, North-West University, 2004. http://hdl.handle.net/10394/411.

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P-glycoprotein (p-gp), a member of the superfamily ATP-binding cassette (ABC) is known to be present in the absorptive enterocytes of the gastro-intestinal tract and many other tissues in the body where it acts mainly as a defence mechanism against exogenous assault. Defects in p-gp is speculated to result in the development of diseases as mutations in genes are causes of numerous diseases in the metabolic mosaic that underlies health. Due to the importance of p-gp, particularly in the intestines, mutation of the gene encoding this protein may lead to the presence of unusual compounds, xenobiotics in the body and the urine. It is thought that defective p-gp in the intestine might also lead to the absorption of some metabolites of bacterial origin and residue of digestion which normally would have been refluxed back into the gut by the p-gp. To investigate if defective p-gp may be involved in the manifestation of unusual compounds and organic acids in the urine, inhibition of intestinal p-gp was proposed. Grapefruit juice (GJ), a natural beverage commonly taken by the majority of the populace has been reported to inhibit p-gp activity in the intestine (Spahn-Langguth & Langguth, 2001). Grapefruit juice was administered to healthy subjects in this study and the sugars and organic acids content of the urine sample after administration was analysed and compared with the controls (urine samples taken from the same set of subjects before grapefruit juice administration). These were determined by thin layer chromatography and gas chromatography-mass spectrophotometry respectively. The thin layer chromatography revealed that there was no difference between the concentrations of sugars in the control and samples after the administration of grapefruit juice. This might indicate that the inhibition of p-gp or mutation of the gene encoding p-gp does not result in the presence of sugars in the urine. The analysis of organic acids by gas chromatography-mass spectrophotometry method showed a remarkable difference between the organic acids present in the controls and urine samples after the administration of grapefruit juice as well as their concentrations. The organic acids solely from microbial origin were statistically analysed and the results gave statistically significant increase in these organic acids in the adults. There was no statistically significant increase in the children. In conclusion, this study confirmed that grapefruit juice inhibits p-gp in the intestine and this resulted in the presence of unusual organic acids from microbial origin in the urine of the adults. The presence of some of these organic acids have been indicated in some metabolic disorders and are also known to give rise to toxic effects on brain, liver, muscle and other tissues. There is the need to do more study on p-gp expression in children so that its functional roles and effect of the mutation of the gene encoding this protein can be known.
Thesis (M.Sc. (Pharm.))--North-West University, Potchefstroom Campus, 2004.

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Carpenter, Melinda. « The expression profile of cytoglobin in human fibrotic lung, and the protective role of cytoglobin in hypoxia and oxidative stress in vitro ». Thesis, University of Birmingham, 2010. http://etheses.bham.ac.uk//id/eprint/623/.

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Cytoglobin (CYGB), a novel member of the globin family, has been shown to be upregulated in response to hypoxia, oxidative stress and fibrogenesis. Presented here is evidence of CYGB expression within cells of fibrotic lesions taken from patients with Idiopathic Pulmonary Fibrosis (IPF) and Chronic Obstructive Pulmonary Disease (COPD). CYGB staining was observed in fibroblasts, endothelial cells, type II pneumocytes, type I pneumocytes, haematopoietic stem cells and inflammatory cells, which were identified using cell specific markers. Cell types which express other members of the globin family, including smooth muscle and red blood cells were negative for CYGB. Fibroblasts were consistently positive for CYGB. CYGB expression was consistently positive within the lesion, and more variable at the edge. This study also provides evidence of an increase in CYGB expression in response to hypoxic and oxidative stress in vitro; however there was no evidence of cytoprotection with over expression of CYGB in response to these insults. There is evidence presented here that the increase in CYGB expression with fibrosis previously reported in the literature, is likely to relate to the hypoxic environment of the lesion and the influx of fibroblasts which are consistently CYGB positive. CYGB is likely to have a role in oxygen and redox homeostasis.

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Castaldo, A. M. « JNK SIGNALLING PATHWAY AND ITS IMPLICATION IN RTT SYNDROME : STUDY ON SYNAPTIC PLASTICITY AND MORPHOLOGICAL PROFILE IN HUMAN AND IN MOUSE MODEL ». Doctoral thesis, Università degli Studi di Milano, 2017. http://hdl.handle.net/2434/491878.

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Rett syndrome (RTT) is a rare and progressive neurodevelopmental disorder that occurs in 1:10,000-15,000 females. RTT is characterized by a normal early growth followed by, above all, motor and cognitive regression. RTT is caused by mutations of MECP2 gene, located on X-chromosome and subjected to random inactivation, that generate a very variable phenotype. Methyl-CpG-binding protein-2 (MeCP2) is a transcriptional factor involved in brain connectivity, neural circuits and importantly, in synaptic plasticity and deficits. However, the molecular mechanisms related with these defects are largely unknown. In previous works, we showed that c-Jun N-terminal protein kinase (JNK), a stress-activated kinase, was strictly involved in synaptic dysfunction related to neurodegenerative disease (Alzheimer’s disease and ischemic stroke) and that its specific inhibition, using the cell permeable D-JNKI1 peptide, led to a recovery of dendritic spines structure and to restoration of functionality supported by a rescue of cognitive deficits. We here proved for the fist time that JNK signalling is powerfully activated in RTT mice and acts as a key modulator of synaptic dysfunctions. The MeCP2 tm 1.1 Bird male mice (referred as Mecp2 y/-) were chosen for our evaluation because, despite not presenting mosaicism, they show an early onset and a most severe phenotype within a homogeneity of genetic background. D-JNKI1 treatment (22mg/kg) was administrated for the first time at 3rd week of age with an intraperitoneal injection and repeated after 3 weeks. Well-being and behavioural studies were effectuated with a weekly examination of food and water intake, weight and locomotor abilities. At 7 weeks, mice were sacrificed and tissues were processed for biochemical evaluations. In brain, we found a strong activation of JNK’s preferential target, c-Jun, a nuclear transcription factor. D-JNKI1 chronic treatment improved general mice wellbeing. Treated mice showed a rescue of motor deficits and an improvement of motor coordination, parameters evaluated with Rotarod and Open field behavioural tests. Since RTT is characterized by locomotor impairment, we checked in cerebellum the synaptic dysfunction evaluating AMPA and NMDA receptors levels. Isolating the post-synaptic region, we found that D-JNKI1 treatment rescued receptor levels. Moreover, PSD95 and Shank3 analysis revealed that the decrement of Mecp2 -/y mice was reported to control level thanks to D-JNKI1 treatment. Moreover, our studies were focused on inflammatory pathway triggered by JNK and we found an astrogliosis and a microgliosis activation, completely rescue by D-JNKI1 treatment. We then move to translational medicine to strengthen the JNK pivotal role in RTT by using Human RTT iPSCs. The mutant neuronal-IPSc presented JNK activation while isogenic control neuronal-IPSc did not; furthermore, we found that D-JNKI1 inhibited JNK activity. The results on iPSCs added value to the clinical relevance of the proposed treatment. RTT is a rare and incurable progressive postnatal female neurodegenerative disorder and the manipulation of JNK pathway may represent the development of an innovative strategy to tackle Rett Syndrome. JNK plays had shown a key role in both mice and human mutated neuronal-IPSc and consequently its relevance in clinical study. We now need to better characterize D-JNKI1 effect in female mosaicist model, closer to the human phenotype.

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Botha, Shani. « The cardiovascular profile of HIV–infected South Africans of African descent : a 5–year prospective study / Botha S ». Thesis, North-West University, 2011. http://hdl.handle.net/10394/7321.

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With great appreciation, I would like to accentuate the substantial contributions of the following people who made this project possible: To Dr. CMT Fourie (my supervisor), Prof. JM van Rooyen (my co–supervisor) and Prof. AE Schutte (my co–supervisor) whose gracious advise, patient guidance, commitment and support have enabled me to plan, analyse, interpret and write this project in a scientific manner. It has been an educational experience for me, thank you. To Mr. LS Wyldbore for the language editing of this dissertation. I thank all the participants, researchers, field workers and supporting staff of the PURE study. The financial assistance of the National Research Foundation (DAAD–NRF) towards this research is hereby acknowledged. A special thanks to my parents, sister, Albert, family and friends, thank you for the never–ending love, support, patience and understanding that you gave me throughout this project. Last, but not the least, a special thank to God for giving me the opportunity, talent, determination and endurance to complete this project.
Thesis (M.Sc. (Physiology))--North-West University, Potchefstroom Campus, 2012.

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Basei, Fernanda Luisa 1983. « Caracterização do perfil de interação da proteína humana Nek4 e sua contextualização funcional = Characterization of the protein interaction profile of the human kinase Nek4 and assignment of its functional context ». [s.n.], 2014. http://repositorio.unicamp.br/jspui/handle/REPOSIP/314361.

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Orientador: Jörg Kobarg
Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
Made available in DSpace on 2018-08-25T14:15:32Z (GMT). No. of bitstreams: 1 Basei_FernandaLuisa_D.pdf: 43645148 bytes, checksum: 4cb7da11417bc57aea32c2db81dddc18 (MD5) Previous issue date: 2014
Resumo: As Neks são proteínas quinases similares a NIMA, proteína que é indispensável para a entrada em mitose de células de Aspergillus nidulans. Em humanos foram identificadas 11 Neks (1-11) e, estudos crescentes vêm demonstrando a participação destas em diversas funções celulares além do controle do ciclo e divisão celular. A Nek4 é um dos maiores membros dessa família e sua relação com a manutenção ciliar e resposta ao DNA danificado já foi demonstrada. Contudo, seus parceiros de interação e substratos são ainda desconhecidos. Para melhor compreender o papel da Nek4 foi realizado um estudo de interatoma para identificar novos processos biológicos com os quais a Nek4 está envolvida. Inicialmente foi identificada uma nova isoforma para a Nek4 e assim, realizou-se o estudo de interatoma para as duas isoformas com a finalidade de comparar o perfil de interação das duas proteínas. As duas isoformas da Nek4 foram expressas em células humanas e após imunoprecipitação seguida de identificação por espectrometria de massas, foram identificadas 474 proteínas que interagem com a isoforma 1 da Nek4, Nek4.1 e 149 para a isoforma 2, Nek4.2. Dentre as proteínas identificadas, 102 interagem com ambas isoformas da Nek4. Nossos resultados confirmam o envolvimento da Nek4 com a resposta ao DNA danificado, função ciliar, estabilização dos microtúbulos e ainda sugerem o envolvimento da Nek4 em funções completamente novas, como processamento de RNAm, resposta ao estresse, controle de qualidade das proteínas e apoptose. Entre os parceiros de interação encontramos importantes proteínas como TRAP-1, Whirlin, PCNA, 14-3-3?, Btf, PARP-1, SRSF1, PAI-RBP1 e KAP-1. As duas isoformas compartilham funções que não foram ainda descritas para os membros da família Nek e a isoforma 1 ainda apresenta funções que já foram descritas para outros membros da família. Aliado ao resultado da imunoprecipitação ainda foram realizadas imunofluorescências que permitiram verificar a localização da Nek4 em diferentes estruturas celulares, como os speckles nucleares e a mitocôndria, corroborando com a função no processamento de RNAm e apoptose. O experimento de imunoprecipitação seguido de identificação por espectrometria de massas também apontou para a possibilidade de autofosforilação e dimerização da Nek4. Além disso, foi possível observar diferenças entre o perfil de interação das duas isoformas da Nek4, sendo que a isoforma 1 interage com proteínas que mantém funções biológicas similares a outras Neks, que a isoforma 2 não apresenta
Abstract: Neks are serine-threonine kinases that are similar to NIMA, a protein found in Aspergillus nidulans which is essential for cell division. In humans there are eleven Neks (1-11) which are involved in different biological functions besides the cell cycle control. Nek4 is one of largest members of the Neks family and has been related to the primary cilia formation and in DNA damage response. However, its substrates and interaction partners are still unknown. Thus in an attempt to better understand the role of Nek4 we performed an interactomics study to find new biological processes in which Nek4 is involved. Besides, we described here a novel Nek4 isoform and compared the interactomics profile of these two Nek4 proteins. Isoform 1 and isoform 2 of Nek4 were expressed in human cells and after an immunoprecipitation followed by mass spectrometry, 474 interacting proteins were identified for isoform 1 and 149 for isoform 2 of Nek4. 102 proteins are common interactors between both isoforms. Our results confirm Nek4 involvement in the DNA damage response, cilia maintenance and microtubules stabilization, and raise the possibility of new functional contexts including mRNA processing, apoptosis signaling, stress response, translation and protein quality control. Among the interaction partners, we found important proteins such as TRAP-1, Whirlin, PCNA, 14-3-3?, Btf, PARP-1, SRSF1, PAI-RBP1 and KAP-1. We could observe that both isoforms share functions that are new to the Nek family, and isoform 1 apparently has also maintained functions which have already been established to other Nek family members. From our immunoprecipitation followed by mass spectrometry experiment a possible site for Nek4 autophosphorylation and dimerization was identified. This study provides new insights into Nek4 functions, identifying new interaction partners, localization to new cellular compartment and further suggests an interesting difference between isoform 1 and the novel isoform 2 of Nek4. Nek4 isoform 1 may have maintained similar roles compared to other Neks and these roles are not related to isoform 2
Doutorado
Bioquimica
Doutora em Biologia Funcional e Molecular

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Afzal, Mansoor. « Graph-Based Visualization of Ontology-Based Competence Profiles for Research Collaboration ». Thesis, Tekniska Högskolan, Högskolan i Jönköping, JTH. Forskningsmiljö Informationsteknik, 2012. http://urn.kb.se/resolve?urn=urn:nbn:se:hj:diva-20123.

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Information visualization can be valuable in a wide range of applications, it deals with abstract, non-spatial data and with the representation of data elements in a meaningful form irrespective of the size of the data, because sometimes visualization itself focuses on the certain key aspects of the data in the representation and thus it helps by providing ease for the goal oriented interpretation. Information visualization focuses on providing a spontaneous and deeper level of the understanding of the data. Research collaboration enhances sharing knowledge and also enhances an individual’s talent. New ideas are generated when knowledge is shared and transferred among each other. According to (He et al, 2009) Research collaboration has been considered as a phenomenon of growing importance for the researchers, also it should be encouraged and is considered to be a “good thing” among the researchers. The main purpose of this thesis work is to prepare a model for the competence profile visualization purpose. For this purpose the study of different visualization techniques that exist in the field of information visualization are discussed in this thesis work. The study and discussion about the visualization techniques motivates in selecting appropriate visualization techniques for the visualization of Ontology-based competence profiles for research collaboration purpose. A proof of concept is developed which shows how these visualization techniques are applied to visualize several components of competence profile.

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Bonoli, Laura. « Profiling the molecular mechanisms driving the fate of human B cells in response to vaccination ». Doctoral thesis, Università degli studi di Padova, 2015. http://hdl.handle.net/11577/3424514.

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Antigen (Ag) encounter activates B cells to proliferate and mature through the formation of germinal centers. Here somatic hypermutation of the variable regions and Immunolgobulin (Ig) isotype switching lead the high affinity Ag-specific clones to two possible differentiation outcomes: antibody (Ab) secreting plasmablasts (PB) or quiescent memory B cells (MBC). The molecular mechanism that drives the fate of a human B cell to differentiate into PB or MBC is poorly understood. Recent studies have provided new insights into the transcriptional program responsible for B cell maturation in mice or human bulk populations. The limited availability of samples and the difficulties in isolating Ag-specific MBCs from peripheral blood make this analysis particularly challenging in humans. We collected samples from human donors that received the seasonal influenza vaccine; those were processed and sorted immediately after the bleed at two different time points: day 8 and day 22 post vaccination, namely the peaks of PBs and MBCs response respectively. The blood samples were used to collect PBs, Ag-specific MBCs and naive B cells (NAIVE) by flow cytometry sorting, exploiting classical surface markers strategies. A new protocol was set up to allow qPCR analysis of multiple genes from sorted single human B cells. This protocol was first used in a pilot study on cells sorted from a first vaccinee, to perform gene expression profiling of 21 relevant genes that allowed us to discriminate the three different B cell populations. Then we up-scaled and optimized the protocol taking advantage of the 96.96 Fluidigm Dynamic Array technology, which enables to perform RT-qPCR for 96 single cells against 96 target genes in one single reaction. This new high-throughput approach was then applied to 240 single cells belonging to Ag-specific MBCs, PBs and NAIVE B cells (80 each) of a second vaccinee, to perform gene expression profiling of 96 genes involved in several pathways of B cell differentiation. By performing unsupervised hierarchical clustering on all the cells, we observed that NAIVE, PBs and MBCs clustered separately and it was possible to identify signatures of gene expression characterizing the three populations. Linear Discriminant Analysis, a dimensionality-reduction analysis, shows that PBs are particularly different from MBCs and NAIVE, that instead share more similarities. By performing statistical analysis we identified the significant differentially expressed genes, which include genes involved in known B cell expression networks and, interestingly, also novel observations (FOXP1, POU2AF1, IRF2). We then compared the gene expression profile of Ag-specific MBCs with MBCs isolated from a healthy donor, to investigate possible differences in the expression patterns of recently activated MBCs and steady-state MBCs. With this analysis we identified 16 genes with a significant differential expression level, denoting a more active profile for the recently activated MBCs isolated from the vaccinee. To further investigate the heterogeneity of Ag-specific MBCs we also recovered immunoglobulin VH sequences from the same cells by sequencing the specific PCR products. Correlation studies showed only weak association between B cell receptor (BCR) maturation (in terms of VH mutation rate) and gene expression data. Conversely, significant association was found between the expression of two genes and the Ig isotype. In particular RORα is associated with IgA, while TBX21 with IgG, in accordance to previous studies performed on mouse bulk B cell populations. The genes identified with this study could be further investigated as they represent potential markers of B cell response to human vaccination.
Nell’ambito del processo di attivazione dovuto all’interazione con l’antigene (Ag), le cellule B proliferano e iniziano un processo di maturazione terminale attraverso la formazione dei centri germinativi (GC). All’interno dei GC, a seguito dell’ipermutazione somatica delle regioni variabili del recettore delle cellule B (BCR) ed il cambiamento di isotipo delle immunoglobuline, i cloni che hanno raggiunto alta affinità per l’Ag possono andare incontro a due possibili destini: differenziamento in plasmablasti (PB) che secernono anticorpi (Ab) o in cellule B della memoria quiescenti (MBC). Il meccanismo molecolare che determina il destino delle cellule B umane durante il differenziamento tardivo in PB o MBC è poco conosciuto. Studi recenti hanno rivelato nuovi aspetti del programma trascrizionale responsabile della maturazione di cellule B in topo o in popolazioni di cellule umane, ma la disponibilità limitata di campioni e la difficoltà nell'isolamento di MBC Ag-specifiche da sangue periferico hanno reso l’analisi di questi tipi cellulari particolarmente complicata. Per questo sono stati raccolti campioni di sangue da donatori sottoposti a vaccinazione stagionale contro l’influenza. Questi campioni sono stati processati immediatamente dopo il prelievo, effettuato in corrispondenza di due particolari momenti: 8 e 22 giorni dopo la vaccinazione, rispettivamente picchi della risposta mediata da PB e da MBC . I campioni di sangue periferico sono stati usati per l’isolamento di PB, NAIVE e MBC Ag-specifiche sfruttando marcatori di superficie. Per l’analisi del profilo di espressione genica è stato ottimizzato un metodo che permette di effettuare qPCR di numerosi geni in cellule B umane isolate come singola cellula. Tale approccio è stato usato inizialmente per uno studio pilota dell’espressione di 21 geni di interesse su cellule isolate da un primo soggetto, permettendoci di discriminare cellule appartenenti alle tre diverse popolazioni. Successivamente questo protocollo è stato ottimizzato sfruttando la tecnologia del 96.96 Dynamic Array prodotto da Fluidigm, sistema che permette di effettuare RT-qPCR su 96 singole cellule per 96 geni in una singola reazione. Con questo metodo ad alta resa abbiamo analizzato 240 singole cellule appartenenti alle popolazioni di MBC Ag-specifiche, PB e NAIVE (80 cellule ciascuna) di un secondo soggetto, permettendoci di analizzare il profilo di espressione di 96 geni coinvolti nelle vie di differenziamento delle cellule B. Attraverso un’analisi statistica di raggruppamento gerarchico dei dati di espressione appartenenti a tutti i campioni processati, abbiamo riunito sotto tre gruppi diversi per espressione genica le cellule appartenenti alle tre diverse popolazioni e abbiamo identificato i geni che le caratterizzano. La Linear Discriminant Analysis, una tecnica di riduzione dimensionale supervisionata, sottolinea come i PB siano particolarmente differenti da MBC Ag-specifiche e NAIVE, che invece condividono un profilo più simile. Sfruttando diversi metodi di analisi statistica, sono stati identificati i geni significativamente espressi in maniera diversa tra le tre popolazioni. Così facendo sono stati individuati sia geni il cui ruolo nella maturazione delle cellule B è noto, sia geni conosciuti principalmente per la loro funzione in altri processi o altre fasi dello sviluppo di queste cellule (FOXP1, POU2AF1, IRF2). Inoltre abbiamo confrontato i profili di espressione delle MBC Ag-specifiche con MBC isolate da un donatore sano non vaccinato, per identificare possibili differenze nei profili di espressione di MBC recentemente attivate e MBC circolanti, lontane dall'attivazione Ag-specifica. Tale analisi ha identificato 16 geni espressi differentemente in maniera significativa, evidenziando un profilo di espressione che denota uno stato di attivazione per le MBC recentemente contattate dall'Ag. Per studiare ulteriormente l’eterogeneità delle MBC Ag-specifiche, tramite PCR abbiamo amplificato e sequenziato le regioni variabili delle catene pesanti (VH) delle immunoglobuline espresse dalle stesse cellule, ma gli studi di correlazione mostrano solo deboli associazioni tra maturazione del BCR (in termini di tasso di mutazione delle VH) e dati di espressione genica. Al contrario, è stata individuata associazione significativa tra la selezione dell'isotipo del BCR e l’espressione di due geni, in particolare l’espressione di RORα è associata alla classe IgA, mentre TBX21 all’IgG, in accordo con studi precedenti effettuati in popolazioni di cellule B murine. In conclusione, i geni identificati da questo studio come discriminanti delle MBC recentemente attivate dall'Ag potrebbero essere ulteriormente studiati in qualità di potenziali marker della risposta B alla vaccinazione in uomo.

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VOLPE, ELISABETTA. « Gene expression profiling of mycobacterium tuberculosis and human macrophage during host-pathogen interaction ». Doctoral thesis, Università degli Studi di Roma "Tor Vergata", 2004. http://hdl.handle.net/2108/208538.

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I macrofagi giocano un ruolo essenziale nella risposta immune a Mycobacterium tuberculosis (Mtb), ma Mtb ha evoluto una serie di meccanismi per superare le risposte macrofagiche e può sopravvivere a lungo all’interno del macrofago umano. Lo studio della risposta trascrizionale all’infezione sia dell’ospite che del patogeno, potrebbe essere interessante per conoscere meglio questa interazione. Con questo scopo noi abbiamo analizzato, a livello trascrizionale, la relazione tra virulento Mtb e macrofago umano dopo 7 giorni di infezione, usando la tecnica del macroarray. Noi abbiamo settato una procedura sperimentale per arricchire di RNA micobatterico, l’RNA totale estratto dalle cellule infettate. Inoltre abbiamo ottimizzato il processo di retrotrascrizione per la generazione del cDNA di Mtb, usando primers specifici per il batterio. Quindi abbiamo analizzato le alterazione di Mtb intracellulare rispetto a Mtb cresciuto in un terreno di coltura sintetico, usando un macroarray con l’intero genoma di Mtb, e abbiamo studiato il profilo di espressione genica dei macrofagi infettati, rispetto ai non infettati, usando un macroarray contenente 858 geni umani coinvolti in processi immunoregolatori. L'analisi globale del trascrittoma di Mtb descritta in questo studio evidenzia un batterio che sente attivamente l’ambiente che lo circonda e che si adatta alle condizioni ostili intracellulari. Dal punto di vista del macrofago, invece, l’infezione determina un’up-regolazione di geni codificanti principalmente molecole con ruolo chemotattico, indicando che il macrofago umano mantiene, dopo 7 giorni di interazione col patogeno, la propria capacità di reclutare altre cellule al sito di infezione. I dati di alcuni geni ottenuti dall’ array sono stati confermati in real-time polymerase chain reactions (PCR). In questo contesto noi abbiamo sviluppato un saggio SYBR Green real-time PCR più specifico e sensibile per rilevare mRNAs micobatterici, rari e ricchi in GC, da campioni di cellule infettate.
Macrophages play an essential role in the immune response to Mycobacterium tuberculosis (Mtb), but Mtb evolved effective mechanisms to survive most macrophage effector functions and it can persist within macrophage longtime. The study of transcriptional response to infection of both host and pathogen, should be interesting to better understand this interaction. To this aim we analyzed, at transcriptional level, the relationship of virulent Mtb and human macrophages after 7 days of infection by macroarrays technique. We set up the experimental procedure to enrich in mycobacterial RNA the total RNA extracted from Mtb-infected cells. We optimized also, the reverse transcription process for Mtb cDNA generation, using Mtb specific primers. Then, we studied simultaneously the gene expression profile of the host and the pathogen. We analyzed the alterations in intracellular Mtb, respect to Mtb grown in synthetic medium, using a macroarray with the whole Mtb genome and the gene expression profile of infected macrophages, versus uninfected ones, using a macroarray containing 858 human genes involved in immunoregulation. The global Mtb transcriptome described in this study suggests an intracellular Mtb that actively sense and adapt itself to hostile environment. On the other hand, human macrophages up-regulate, mainly, genes encoding for molecules with a chemotactic role, indicating their maintenance of capacity to recruit other cells at the site of infection, after 7 days of interaction with the pathogen. The data for a selected group of the modulated genes were confirmed by real-time polymerase chain reactions (PCR). In this context we developed a more sensitive and more specific SYBR Green real-time PCR assay to detect rare and GC-rich mycobacterial mRNAs from infected samples.

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MANCINO, ROBERTA. « Influence of cow diet on nutritional profile of milk and dairy products and effects on alterations of human gut microbiota by an in vitro digestion model ». Doctoral thesis, Università di Foggia, 2018. http://hdl.handle.net/11369/363264.

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I consumatori richiedono costantemente la presenza sul mercato di alimenti salutistici, che in termini di latte e prodotti lattiero-caseari si traducono in un latte con percentuali più elevate di acidi grassi “sani” come gli acidi grassi polinsaturi (PUFA), ed una più bassa percentuale di acidi grassi saturi (SFA). Il latte e prodotti lattiero-caseari contribuiscono in modo significativo all’assorbimento di sostanze nutritive essenziali nella dieta. Nonostante ciò, il consumo di latte ha fatto registrare un’inflessione a causa delle linee guida nutrizionali che suggeriscono di limitare il consumo pro capite di SFA, che per una parte significativa provengono da latte e derivati (USDA e HHS, 2010). Una strategia per migliorare il profilo di FA di latte e prodotti lattiero-caseari è l'integrazione della dieta bovina con semi oleaginosi, che diminuiscono la percentuale di SFA nel latte, diminuendo la sintesi de novo di FA nella ghiandola mammaria. Da precedenti sperimentazioni emerge che la supplementazione di semi di lino nella dieta delle bovine da latte riduce le concentrazioni di acidi grassi a corta catena corta e acidi grassi a media catena, ed aumenta il contenuto di acidi grassi a lunga catena nel grasso del latte. Tuttavia, l’inclusione dei semi oleaginosi, in particolare dei semi di lino, nella dieta della vacca da latte, scoraggia gli allevatori dal loro utilizzo, a causa del costo molto elevato. É necessario, quindi, trovare un compromesso tra costi aggiuntivi stimati e la giusta quantità di semi oleaginosi da somministrare nella dieta degli animali per migliorare la produzione di latte e la sua composizione. In Italia, circa l'80% delle aziende lattiero-casearie utilizza vacche di razza Frisona per la produzione lattea da destinare sia al consumo diretto che alla produzione di formaggio. Le bovine di razza Jersey vengono utilizzate per migliorare l'efficienza del settore della produzione di formaggio in diverse parti del mondo. Il tratto gastrointestinale costituisce la più grande interfaccia del corpo con l'ambiente esterno ed è esposto ad una grande quantità di materiale estraneo, inclusi batteri patogeni e commensali, così come antigeni alimentari. La tolleranza orale è una proprietà importante del sistema immunitario intestinale; l’omeostasi intestinale richiede interazioni equilibrate tra il microbiota intestinale e gli antigeni alimentari. Alla nascita, siamo colonizzati da una complessa comunità di cellule mcrobiche che arriva fino a una densità di 1 × 1012 cellule batteriche per grammo di microbi contenuti nel colon adulto. Queste cellule microbiche vivono in una relazione simbiotica con l'ospite e sono determinanti in materia di salute e di malattia, influenzando l'assorbimento dei nutrienti, la funzione di barriera e lo sviluppo del sistema immunitario. Sulla base delle considerazioni precedenti gli scopi del presente lavoro di tesi sono: 1. cercare di ridurre la quantità giornaliera di semi di lino somministrati agli animali per incoraggiare il loro utilizzo da parte degli allevatori per aumentare il contenuto di acidi grassi polinsaturi nel latte a scapito degli acidi grassi saturi con una connotazione salutistica del latte prodotto; 2. testare gli effetti della somministrazione di semi di lino su due diverse razze da latte: Frisona e Jersey; 3. valutare il trasferimento di acidi grassi polinsaturi in due diversi prodotti lattiero-caseari (Caciotta vs Caciocavallo) con tempi di stagionatura diversi; 4. valutare gli effetti dei prodotti lattiero-caseari naturalmente arricchiti in acidi grassi polinsaturi sulla salute umana di un modello di digestione in vitro con la valutazione di: a) variazioni di profilo degli acidi grassi di prodotti lattiero-caseari dopo la digestione in vitro; b) acidi grassi a catena corta (SCFA) prodotti dal gut microbiota; c) variazioni della flora intestinale tramite fermentazione fecale seguito da pirosequenziamento. Dai risultati ottenuti il contenuto elevato di latte C18:3n3 nel latte suggerisce che la riduzione della quantità di supplementazione di semi di lino nella dieta delle bovine può migliorare latte profilo di acidi grassi con una consistente riduzione dei costi di produzione; le razze sottoposet alla sperimentazione, Frisona e Jersey hanno risposto in modo diverso alla stessa supplementazione di semi di lino. Gli acidi grassi polinsaturi sono stati trasferiti nei prodotti lattiero caseari, soprattutto nella Caciotta, suggerendo un ruolo determinante del protocollo di caseificazione nel trasferimento della tipologia di acido grasso della dieta. Dopo la digestione in vitro, gli acidi grassi rimangono nel digerito gastro-intestinale; la loro presenza può avere effetti benefici sul tratto gastrointestinale e di conseguenza sulla salute umana. Inoltre la presenza e la quantità di acidi grassi a catena corta (SCFA) ritrovata nei fermentati fecali suggeriscono alcuni cambiamenti delle popolazioni microbiche indotte dalla presenza dei prodotti lattiero caseari sperimentali, lasciando intravedere importanti effetti benefici sulla salute umana.
Health-conscious consumers are demanding milk with higher proportions of healthy fatty acids as polyunsatured fatty acids (PUFA), and lower proportion of saturated fatty acids (SFA). Milk and dairy products contribute significantly to the consumption of essential nutrients in human populations. Despite its important roles in human nutrition, consumption of milk has declined, because nutritional guidelines have limited capita consumption of SFA, which to a significant proportion originate from milk and dairy products (USDA and HHS, 2010). A strategy to improve the FA profile of milk and dairy products is the supplementation of cow’s diet with oilseeds, which decrease the proportion of SFA, by decreasing de novo FA synthesis in the mammary gland. Feeding flaxseed to dairy cows decreases the concentrations of short-chain fatty acids and medium chain fatty acids and increases the long-chain fatty acid content in milk fat. However, oilseeds, and in particular flaxseed, have a very high costs that discourage farmers in their utilization. It’s necessary, therefore to find a compromise between costs and the right amount to be administered in the diet to the animals to ameliorate milk yield and composition. In Italy, about 80% of dairy farms produce milk of Friesian cows both for direct consumption and for cheese production. Jersey breed and it has been used to improve the efficiency of the cheesemaking sector in different part of the world, and is characterized by improved longevity, superior udder health, higher cheese yield, reduced feed and water requirement. The gastrointestinal tract constitutes the body’s largest interface with the external environment and is exposed to a vast amount of foreign material, including pathogenic and commensal bacteria, as well as food antigens. Oral tolerance is an important property of the gut immune system; intestinal homeostasis requires balanced interactions between the gut microbiota, dietary antigens. At birth, we are colonized with a complex community of microbes that reaches up to a density of 1 × 1012 bacterial cells per grams of content in the adult colon. These microbes live in a symbiotic relationship with the host and they are determinants in health and disease influencing nutrient absorption, barrier function and immune development. On the basis of the previous considerations and considering that oil seeds are expensive and many farmers are reluctant to use them the aims of this PhD thesis are: 1. trying to reduce the daily amount of flaxseed administered to animals in order to increase the content of polyunsaturated fatty acids in milk at the expense of saturated fatty acids, and to encourage its utilization by farmers as supplements to dairy cows with a reduction of management costs; 2. testing the effects of flaxseed administration on two different dairy cows breeds: Friesian and Jersey; 3. evaluating the transferring of polyunsaturated fatty acids in two different dairy products (Caciotta vs Caciocavallo) at different ripening time; 4. evaluating the effects of dairy products naturally enriched in polyunsaturated fatty acids on human health by an in vitro digestion model with the evaluation of changes in: a) fatty acid profile of dairy products after in vitro digestion; b) short chain fatty acids (SCFA) produced by gut microbiota; c) changes in gut microbiota populations by fecal fermentation followed by pyrosequencing. The higher milk content of C18:3n3 in milk suggests that the reduction in the amount of flaxseed supplementation can also improve milk fatty acid profile with a consistent reduction of production costs; however, Friesian and Jersey cows replied differently to the same flaxseed supplementation; Polyunsatured fatty acids are transferred into dairy products, especially in Caciotta cheese, suggesting that probably the different cheese making influenced the transferring. After in vitro digestion, fatty acids remain in the digest; their presence can have beneficial effects on the gastrointestinal tract and consequently on human health. Moreover the presence and the amount of short chain fatty acids (SCFA) could suggest some changes of microbiological populations that could have beneficial effects on human health.

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Danielová, Tereza. « Obrazová analýza mitotických chromosomů ». Master's thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2014. http://www.nusl.cz/ntk/nusl-220846.

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This master’s thesis is focused on digital image analysis of mitotic chromosomes. It deals with the design of the processing of digital images - from image preprocessing to clasification of each chromosomes, including testing on a set of images. This work introduces used cytogenetic methods, that are used to visualize chromosomes. In its practical part describes morphology operations and clasification procedure. Classification of the chomosomes was divided into 5 groups (A-G). All algorithms were created in the MATLAB program.

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