Bone marrow mesenchymal stem cells overexpressing human basic fibroblast growth factor increase vasculogenesis in ischemic rats

Administration or expression of growth factors, as well as implantation of autologous bone marrow cells, promote in vivo angiogenesis. This study investigated the angiogenic potential of combining both approaches through the allogenic transplantation of bone marrow-derived mesenchymal stem cells (MSCs) expressing human basic fibroblast growth factor (hbFGF). After establishing a hind limb ischemia model in Sprague Dawley rats, the animals were randomly divided into four treatment groups: MSCs expressing green fluorescent protein (GFP-MSC), MSCs expressing hbFGF (hbFGF-MSC), MSC controls, and phosphate-buffered saline (PBS) controls. After 2 weeks, MSC survival and differentiation, hbFGF and vascular endothelial growth factor (VEGF) expression, and microvessel density of ischemic muscles were determined. Stable hbFGF expression was observed in the hbFGF-MSC group after 2 weeks. More hbFGF-MSCs than GFP-MSCs survived and differentiated into vascular endothelial cells (P<0.001); however, their differentiation rates were similar. Moreover, allogenic transplantation of hbFGF-MSCs increased VEGF expression (P=0.008) and microvessel density (P<0.001). Transplantation of hbFGF-expressing MSCs promoted angiogenesis in an in vivo hind limb ischemia model by increasing the survival of transplanted cells that subsequently differentiated into vascular endothelial cells. This study showed the therapeutic potential of combining cell-based therapy with gene therapy to treat ischemic disease.

Securin, Cdc20 and Cdc27 in predicting the outcome of human breast cancer

Deregulated proliferation has been recognized among the most important factors promoting breast cancer development and progression. The aim of the project is to gain understanding of the role of specific cell cycle regulators of metaphase-anaphase transition and evaluate their potential in breast cancer prognostication and treatment decisions. Metaphase-anaphase transition is triggered by activation of anaphase promoting complex (APC) which is activated by a cascade of regulatory proteins, among them securin, Cdc20 and Cdc27. These proteins promote the metaphase–anaphase transition and participate in the timely separation of the chromatids. This study is based on a patient material of approximately 600 breast cancer patients and up to 22 years of follow-up. As the main observation, based on DNA cytometric and immunohistochemical methods, securin, Cdc20 and Cdc27 protein expressions were associated with abnormal DNA content and outcome of breast cancer. In the studied patient material, high securin expression alone and in combination with Cdc20 and Cdc27 predicted up to 9.8-fold odds for aneuploid DNA content in human breast cancer. In Kaplan–Meier analyses, high expression of securin systematically indicated decrease in breast cancer survival as compared to low expression cases. The adverse effect of high securin expression was further strengthened by combining it with Cdc20 or Cdc27 expressions, resulting in up to 6.8-fold risk of breast cancer death. High securin and Cdc20 expression was also associated with triple-negative breast cancer type with high statistical significance. Securin, Cdc20 or Cdc27 have not previously been investigated in a clinically relevant large breast cancer patient material or in association with DNA ploidy. The present findings suggest that the studied proteins may serve as potential biomarkers for identification of aggressive course of disease and unfavourable outcome of human breast cancer, and that they may provide a future research aim for understanding abnormal proliferation in malignant disease.

Use of cereal bars with quinoa (Chenopodium quinoa W.) to reduce risk factors related to cardiovascular diseases

Quinoa is considered a pseudocereal with proteins of high biological value, carbohydrates of low glycemic index, phytosteroids, and omega-3 and 6 fatty acids that bring benefits to the human health. The purpose of this study was to investigate the effects of quinoa on the biochemical and anthropometric profile and blood pressure in humans, parameters for measuring risk of cardiovascular diseases. Twenty-two 18 to 45-year-old students were treated daily for 30 days with quinoa in the form of a cereal bar. Blood samples were collected before and after 30 days of treatment to determine glycemic and biochemical profile of the group. The results indicated that quinoa had beneficial effects on part of the population studied since the levels of total cholesterol, triglycerides, and LDL-c showed reduction. It can be concluded that the use of quinoa in diet can be considered beneficial in the prevention and treatment of risk factors related to cardiovascular diseases that are among the leading causes of death in today's globalized world. However, further studies are needed to prove the benefits observed.

Chemical composition of tempeh from soybean cultivars specially developed for human consumption

Tempeh is a food obtained by fermentation of soybean grains by the fungus Rizophus oligosporus. It is a traditional Indonesian food that presents benefits for human health protecting against diarrhea and chronic diseases. Tempeh processing includes dehulling, cooking, inoculation, and fermentation. In this study, chemical characteristics of tempeh prepared with soybean cultivars specially developed for human consumption (BRS 216, BRS 232, BRS 257, and BRS 267) were investigated. Soybean grains and tempeh obtained from these cultivars were analyzed for oil, protein, antinutrional factors, and isoflavone content. Cultivar BRS 216 presented the highest protein content in the grains (36.81%) and in tempeh (51.99%). On average, the protein content in tempeh increased 16% in relation to that of soybean grains. Isoflavone content was higher in the grains than in tempeh with significant differences among the cultivars. However, the aglycones content increased about 50% in tempeh (49.00 mg.100 g-1 on average) compared to that of raw material (soybean grains - 21.49 mg.100 g-1, on average). The content of Kunitz trypsin inhibitor (KSTI) reduced 83% in tempeh, on average, as compared to the value found in the grains. Phytic acid content was similar in both tempeh and the grains.

The importance of heat against antinutritional factors from Chenopodium quinoa seeds

Chenopodium quinoa seeds have high protein content. The nutritional value of quinoa is superior compared with traditional cereals. Its essential amino acid composition is considered next to the ideal, and its quality matches that of milk proteins. In this study, the seed storage proteins from Chenopodium quinoa were extracted, fractionated, partially purified, and characterized. The structural characterization was performed by Tricine-SDS-PAGE and two-dimensional electrophoresis, and it confirmed the presence of proteins of molecular weight of 30 and 7kDa, probably corresponding to lectins and trypsin inhibitors, respectively. The functional characterization of these proteins evidenced their activity as antinutritional factors due to their in vitro digestibility. Quinoa proteins have an excellent amino acid composition with many essential amino acids. In vitro digestibility evaluation indicated that heat-treated samples showed a more complete digestion than the native state samples. Quinoa seeds can be an important cereal in human diet after adequate heat treatment.

A human-centric service approach for architecting and reengineering global system

Production of a new system in any range is expanding dramatically and new ideas are there upon introduced, the logic stands behind the matter is the growth of application of the internet and granting web-based systems. Before producing a system and distribute to the customer, various aspects should be studied which multiple the profit of the system. The process of productizing a new system from being unprocessed idea until delivers to the final user has been unambiguous. In this thesis, the systematize service in a way that benefits both the customer and provider, along with an effort to establish trust and diminish customer’s risk and increase service productivity are in detail presented. Characteristics of Servitization and Productization as two faces of one coin have been interpreted. Apart from the abovementioned issues state of art, service-oriented architecture (SOA) and New Service Development (NSD) has been included in this report for solving the problem of gradually decline in value of companies.

Continental margin architecture : the palynological signature of glacioeustasy /

Palynomorphs from two siliciclastic margins were examined to gain insights into continental margin architecture. Sea level change is thought to be one of the primary controls on continental margin architecture. Because Late Neogene glacioeustasy has been well studied marine sediments deposited during the Late Neogene were examined to test this concept. Cores from the outer shelf and upper slope were taken from the New Jersey margin in the western North Atlantic Ocean and from the Sunda Shelf margin in the South China Sea. Continental margin architecture is often described in a sequence stratigraphic context. One of the main goals of both coring projects was to test the theoretical sequence stratigraphic models developed by a research group at Exxon (e.g. Wilgus et al., 1988). Palynomorphs provide one of the few methods of inferring continental margin architecture in monotonous, siliciclastic marine sediments where calcareous sediments are rare (e.g. New Jersey margin). In this study theoretical models of the palynological signature expected in sediment packages deposited during the various increments of a glacioeustatic cycle were designed. These models were based on the modem palynomorph trends and taphonomic factors thought to control palynomorph distribution. Both terrestrial (pollen and spores) and marine (dinocysts) palynomorphs were examined. The palynological model was then compared with New Jersey margin and Sunda Shelf margin sediments. The predicted palynological trends provided a means of identifying a complete cycle of glacioeustatic change (Oxygen Isotope Stage 5e to present) in the uppermost 80 meters of sediment on the slope at the New Jersey margin. Sediment availability, not sea meters of sediment on the slope at the New Jersey margin. Sediment availability, not sea level change, is thought to be the major factor controlling margin architecture during the late Pleistocene here at the upper slope. This is likely a function of the glacial scouring of the continents which significantly increases sediment availability during glacial stages. The subaerially exposed continental shelf during the lowstand periods would have been subject to significant amounts of erosion fi:om the proglacial rivers flowing fi-om the southern regions of the ice-sheet. The slope site is non-depositional today and was also non-depositional during the last full interglacial period. The palynomorph data obtained fi-om the South China Sea indicate that the major difference between the New Jersey Margin sites and the Sunda Shelf margin sites is the variation in sediment supply and the rate of sediment accumulation. There was significantly less variation in sediment supply between glacial and interglacial periods and less overall sediment accumulation at the Sunda Shelf margin. The data presented here indicate that under certain conditions the theoretical palynological models allow the identification of individual sequence stratigraphic units and therefore, allow inferences regarding continental margin architecture. The major condition required in this approach is that a complete and reliable database of the contemporaneous palynomorphs be available.

Investigation of the mechanism of transfer of a-tocopherol by the human a-tocopherol transfer protein (H-a-TTP) /

The human a-tocopherol transfer protein (h-a-TTP) is understood to be the entity responsible for the specific retention of a-tocopherol (a-toc) in human tissues over all other forms of vitamin E obtained from the diet. a-Tocopherol is the most biologically active form of vitamin E, and to date has been studied extensively with regard to its antioxidant properties and its role of terminating membrane lipid peroxidation chain reactions. However, information surrounding the distribution of a-tocopherol, specifically its delivery to intracellular membranes by a-TTP, is still unclear and the molecular factors influencing transfer remain elusive. To investigate the mechanism of ligand transfer by the h-a-TTP, a fluorescent analogue of a-toc has been used in the development of a fluorescence resonance energy transfer (FRET) assay. (/?)-2,5,7,8-tetramethyl-2-[9-(7-nitro-benzo[l,2,5]oxdiazol-4-ylamino)-nonyl]- chroman-6-ol (NBD-toc) has allowed for the development of the FRET-based ligand transfer assay. This ligand has been utilized in a series of experiments where changes were made to acceptor lipid membrane concentration and composition, as well as to the ionic strength and viscosity of the buffer medium. Such changes have yielded evidence supporting a collisional mechanism of ligand transfer by a-TTP, and have brought to light a new line of inquiry pertaining to the nature of the forces governing the collisional transfer interaction. Through elucidation of the transfer mechanism type, a deeper understanding of the transfer event and the in vivo fate of a-tocopherol have been obtained. Furthermore, the results presented here allow for a deeper investigation of the forces controlling the collisional protein-membrane interaction and their effect on the transfer of a-toc to membranes. Future investigation in this direction will raise the possibility of a complete understanding of the molecular events surrounding the distribution of a-toc within the cell and to the body's tissues.

The capillary supply of human skeletal muscle in health and disease

BACKGROUND: Capillaries function to provide a surface area for nutrient and waste exchange with cells. The capillary supply of skeletal muscle is highly organized, and therefore, represents an excellent choice to study factors regulating diffusion. Muscle is comprised of three specific fibre types, each with specific contractile and metabolic characteristics, which influence the capillary supply of a given muscle; in addition, both environmental and genetic factors influence the capillary supply, including aging, physical training, and various disease processes. OBJECTIVE: The present study was undertaken to develop and assess the functionality of a data base, from which virtual experiments can be conducted on the capillary supply of human muscle, and the adaptations of the capillary bed in muscle to various perturbations. METHODS: To create the database, an extensive search of the literature was conducted using various search engines, and the three key words - "capillary, muscle, and human". This search yielded 169 papers from which the data for the 46 variables on the capillary supply and fibre characteristics of muscle were extracted for inclusion in the database. A series of statistical analyses (ANOVA) were done on the capillary database to examine differences in skeletal muscle capillarization and fibre characteristics between young and old individuals, between healthy and diseased individuals, and between untrained, endurance trained, endurance welltrained, and resistance trained individuals, using SAS. RESULTS: There was a significantly higher capillarization in the young compared to the old individuals, in the healthy compared to the diseased individuals, and in the endurance-trained and endurance well-trained compared to the untrained individuals. CONCLUSIONS: The results of this study support the conclusion that the capillary supply of skeletal muscle is closely regulated by factors aimed at optimizing oxygen and nutrient supply and/or waste removal in response to changes in muscle mass and/or metabolic activity.

The Importance and achievement of work values and locus of control as factors in work motivation

The present research study examined the relationships in a work motivation context among perceived importance and achievement of work values, locus of control and internal work motivation. The congruence of a work value was considered to be the discrepancy between the importance of a work value and the perceived achievement of that value. The theoretical framework utilized was based on a self-perpetuating cycle of motivation which included the perceived importance and achievement of work values and internal work motivation as separate and distinct, yet interrelated factors. It was hypothesized that individuals who experienced high congruence of work values would experience higher levels of internal work motivation than individuals who had low congruence of work values. It was also hypothesized that individuals who had an internal locus of control would experience more internal work motivation individuals well, the and have higher congruence of work values than who had an external locus of control. As possibility of locus of control as a moderator between importance of work values and internal work motivation was explored. Survey data were collected from 184 managerial level employees of the XYZ company during an ongoing training session. The following instruments were employed to measure the variables: Elizur's (1984) Importance of Work Values, Hunt and Saul's (1985) Achievement of Work Values, Hatfield, Robinson and Huseman's (1975) Job Perception Scale, a modified version of Rotter's (1966) I-E Locus of Control Scale and the Internal Work Motivation Scale (Hackman & Oldham, 1980) which is a part of the Job Diagnostic Survey. The findings indicated that locus of control was not a significant factor in determining congruence between work values or internal work motivation for this sample. Furthermore, locus of control was also found not to be a moderator between the importance of work values and internal work motivation. All individuals in this study had relatively high levels of internal work motivation. However, individuals who had higher congruence of work values did have significantly higher internal work motivation than those who had low congruence of work values for a majority of the 21 values. This was particularly true for the intrinsic values which included responsibility, meaningfulness and use of abilities. In addition, the data were analysed into a hierarchy of needs to indicate possible organizational development or human resource development needs for the XYZ corporation.

Taenia solium transmission in a rural community in Honduras : an examination of risk factors and knowledge

Taenia soliurn taeniasis and cysticercosis are recognized as a major public health problem in Latin America. T. soliurn transmission not only affects the health of the individual, but also social and economic development, perpetuating the cycle of poverty. To determine prevalence rates, population knowledge and risk factors associated with transmission, an epidemiological study was undertaken in the rural community of Jalaca. Two standardized questionnaires were used to collect epidemiological and T. soli urn general knowledge data. Kato-Katz technique and an immunoblot assay (EITB) were used to determine taeniasis and seroprevalence, respectively. In total, 139 individuals belonging to 56 households participated in the study. Household characteristics were consistent with conditions of poverty of rural Honduras: 21.4% had no toilet or latrines, 19.6% had earthen floor, and 51.8% lacked indoor tap water. Pigs were raised in 46.4% of households, of which 70% allowed their pigs roaming freely. A human seroprevalence rate of 18.7% and a taeniasis prevalence rate of 2.4% were found. Only four persons answered correctly 2: 6 out of ten T. soliurn knowledge questions, for an average passing score of 2.9%. In general, a serious gap exists in knowledge regarding how humans acquire the infections, especially neurocysticercosis was identified. After regression analysis, the ability to recognize adult tapeworms and awareness of the clinical importance of taeniasis, were found to be significant risk factors for T. soliurn seropositivity. These results demonstrate a high level of transmission and a low level of kn~,wledge about Taenia soliurn in Jalaca. Consequently, intervention measures integrated with health education are necessary to decrease the burden caused by this parasite.

"I might be a duck, but I'm human" : an analysis of clothing in Disney cartoons

Mickey Mouse, one of the world's most recognizable cartoon characters, did not wear a shirt in his earliest incarnation in theatrical shorts and, for many years, Donald Duck did not wear pants and still rarely does so. Especially when one considers the era in which these figures were first created by the Walt Disney Studio, in the 1920s and 1930s, why are they portrayed without full clothing? The obvious answer, of course, is that they are animals, and animals do not wear clothes. But these are no ordinary animals: in most cases, they do wear clothing - some clothing, at least - and they walk on two legs, talk in a more or less intelligible fashion, and display a number of other anthropomorphic traits. If they are essentially animals, why do they wear clothing at all? On the other hand, if these characters are more human than animal, as suggested by other behavioral traits - they walk, talk, work, read, and so on - why are they not more often fully clothed? To answer these questions I undertook three major research strategies used to gather evidence: interpretive textual analysis of 321 cartoons; secondary analysis of interviews conducted with the animators who created the Disney characters; and historical and archival research on the Disney Company and on the times and context in which it functioned. I was able to identify five themes that played a large part in what kind of clothing a character wore; first, the character's gender and/or sexuality; second, what species or "race" the character was; third, the character's socio-economic status; fourth, the degree to which the character was anthropomorphized; and, fifth, the context in which the character and its clothing appeared in a particular scene or narrative. I concluded that all of these factors played a part in determining, to some extent, the clothing worn by particular characters at particular times. However, certain patterns emerged from the analysis that could not be explained by these factors alone or in combination. Therefore, my analysis also investigates the individual and collective attitudes and desires of the men in the Disney studio who were responsible for creating these characters and the cultural conditions under which they were created. Drawing on literature from the psychoanalytic approach to film studies, I argue that the clothing choices spoke to an idealized fantasy world to which the animators (most importantly, Walt Disney himself), and possibly wider society, wanted to return.

HUMAN GENOME VARIATIONS AND EVOLUTION WITH A FOCUS ON THE ANALYSIS OF TRANSPOSABLE ELEMENTS

Genome sequence varies in numerous ways among individuals although the gross architecture is fixed for all humans. Retrotransposons create one of the most abundant structural variants in the human genome and are divided in many families, with certain members in some families, e.g., L1, Alu, SVA, and HERV-K, remaining active for transposition. Along with other types of genomic variants, retrotransponson-derived variants contribute to the whole spectrum of genome variants in humans. With the advancement of sequencing techniques, many human genomes are being sequenced at the individual level, fueling the comparative research on these variants among individuals. In this thesis, the evolution and functional impact of structural variations is examined primarily focusing on retrotransposons in the context of human evolution. The thesis comprises of three different studies on the topics that are presented in three data chapters. First, the recent evolution of all human specific AluYb members, representing the second most active subfamily of Alus, was tracked to identify their source/master copy using a novel approach. All human-specific AluYb elements from the reference genome were extracted, aligned with one another to construct clusters of similar copies and each cluster was analyzed to generate the evolutionary relationship between the members of the cluster. The approach resulted in identification of one major driver copy of all human specific Yb8 and the source copy of the Yb9 lineage. Three new subfamilies within the AluYb family – Yb8a1, Yb10 and Yb11 were also identified, with Yb11 being the youngest and most polymorphic. Second, an attempt to construct a relation between transposable elements (TEs) and tandem repeats (TRs) was made at a genome-wide scale for the first time. Upon sequence comparison, positional cross-checking and other relevant analyses, it was observed that over 20% of all TRs are derived from TEs. This result established the first connection between these two types of repetitive elements, and extends our appreciation for the impact of TEs on genomes. Furthermore, only 6% of these TE-derived TRs follow the already postulated initiation and expansion mechanisms, suggesting that the others are likely to follow a yet-unidentified mechanism. Third, by taking a combination of multiple computational approaches involving all types of genetic variations published so far including transposable elements, the first whole genome sequence of the most recent common ancestor of all modern human populations that diverged into different populations around 125,000-100,000 years ago was constructed. The study shows that the current reference genome sequence is 8.89 million base pairs larger than our common ancestor’s genome, contributed by a whole spectrum of genetic mechanisms. The use of this ancestral reference genome to facilitate the analysis of personal genomes was demonstrated using an example genome and more insightful recent evolutionary analyses involving the Neanderthal genome. The three data chapters presented in this thesis conclude that the tandem repeats and transposable elements are not two entirely distinctly isolated elements as over 20% TRs are actually derived from TEs. Certain subfamilies of TEs themselves are still evolving with the generation of newer subfamilies. The evolutionary analyses of all TEs along with other genomic variants helped to construct the genome sequence of the most recent common ancestor to all modern human populations which provides a better alternative to human reference genome and can be a useful resource for the study of personal genomics, population genetics, human and primate evolution.

Systematic analysis of protein complexes involved in the human RNA polymerase II machinery

La transcription, la maturation d’ARN, et le remodelage de la chromatine sont tous des processus centraux dans l'interprétation de l'information contenue dans l’ADN. Bien que beaucoup de complexes de protéines formant la machinerie cellulaire de transcription aient été étudiés, plusieurs restent encore à identifier et caractériser. En utilisant une approche protéomique, notre laboratoire a purifié plusieurs composantes de la machinerie de transcription de l’ARNPII humaine par double chromatographie d’affinité "TAP". Cette procédure permet l'isolement de complexes protéiques comme ils existent vraisemblablement in vivo dans les cellules mammifères, et l'identification de partenaires d'interactions par spectrométrie de masse. Les interactions protéiques qui sont validées bioinformatiquement, sont choisies et utilisées pour cartographier un réseau connectant plusieurs composantes de la machinerie transcriptionnelle. En appliquant cette procédure, notre laboratoire a identifié, pour la première fois, un groupe de protéines, qui interagit physiquement et fonctionnellement avec l’ARNPII humaine. Les propriétés de ces protéines suggèrent un rôle dans l'assemblage de complexes à plusieurs sous-unités, comme les protéines d'échafaudage et chaperonnes. L'objectif de mon projet était de continuer la caractérisation du réseau de complexes protéiques impliquant les facteurs de transcription. Huit nouveaux partenaires de l’ARNPII (PIH1D1, GPN3, WDR92, PFDN2, KIAA0406, PDRG1, CCT4 et CCT5) ont été purifiés par la méthode TAP, et la spectrométrie de masse a permis d’identifier de nouvelles interactions. Au cours des années, l’analyse par notre laboratoire des mécanismes de la transcription a contribué à apporter de nouvelles connaissances et à mieux comprendre son fonctionnement. Cette connaissance est essentielle au développement de médicaments qui cibleront les mécanismes de la transcription.

Globin Gene Expression: Role of Transcription Factors

La dérégulation de l'expression génétique est une base pathophysiologique de plusieurs maladies. On a utilisé le locus du gène β-globine humain comme modèle pour élucider le mécanisme de régulation de la transcription génétique et évaluer son expression génétique durant l'érythropoïèse. La famille des protéines 'E' est composée de facteurs de transcription qui possèdent plusieurs sites de liaison au sein de locus du gène β-globine, suggérant leur rôle potentiel dans la régulation de l’expression de ces gènes. Nous avons montré que les facteurs HEB, E2A et ETO2 interagissent d’une manière significative avec la région contrôle du Locus (LCR) et avec les promoteurs des gènes de la famille β-globine. Le recrutement de ces facteurs au locus est modifié lors de l'érythropoïèse dans les cellules souches hematopoitiques et les cellules erythroides de souris transgéniques pour le locus de la β-globine humain, ainsi que dans les cellules progénitrices hématopoïétiques humaines. De plus par cette étude, nous démontrons pour la première fois que le gène β-globine humain est dans une chromatine active et qu’il interagit avec des facteurs de transcriptions de type suppresseurs dans les cellules progénitrices lymphoïdes (voie de différentiation alternative). Cette étude a aussi été faite dans des souris ayant une génétique mutante caractérisée par l'absence des facteurs de transcription E2A ou HEB.