Role of ErbB2 and ErbB4 in Cancer Growth, Prognosis and as Targets for Immunotherapy

ErbB receptors (EGFR, ErbB2, ErbB3 and ErbB4) are growth factor receptors that regulate signals of cell differentiation, proliferation, migration and survival. Inappropriate activation of these receptors is associated with the development and severity of many cancers and has prognostic and predictive value in cancer therapy. Drugs, such as therapeutic antibodies, targeted against EGFR and ErbB2, are currently used in therapy of breast, colorectal and head and neck cancers. The role of ErbB4 in tumorigenesis has remained relatively poorly understood. Alternative splicing produces four different isoforms of one ErbB4 gene. These isoforms (JM-a, JM-b, CYT-1 and CYT-2) are functionally dissimilar and proposed to have different roles in carcinogenesis. The juxtamembrane form JM-a undergoes regulated intramembrane proteolysis producing a soluble receptor ectodomain and an intracellular domain that translocates into the nucleus and regulates transcription. Nuclear signaling via JM-a isoform stimulates cancer cell proliferation. This study aimed to develop antibodies targeting the proposed oncogenic ErbB4 JM-a isoform that show potential in inhibiting ErbB4 dependent tumorigenesis. Also, the clinical relevance of ErbB4 shedding in cancer was studied. The currently used monoclonal antibody trastuzumab, targeting ErbB2, has shown efficacy in breast cancer therapy. In this study novel tissues with ErbB2 amplification and trastuzumab sensitivity were analyzed. The results of this study indicated that a subpopulation of breast cancer patients demonstrate increased shedding and cleavage of ErbB4. A JM-a isoform-specific antibody that inhibited ErbB4 shedding and consequent activation of ErbB4 had anti-tumor activity both in vitro and in vivo. Thus, ErbB4 shedding associates with tumor growth and specific targeting of the cleavable JM-a isoform could be considered as a strategy for developing novel ErbB-based cancer drugs. In addition, it was demonstrated that ErbB2 amplification is common in intestinal type gastric cancers with poor clinical outcome. Trastuzumab inhibited growth of gastric and breast cancer cells with equal efficacy. Thus, ErbB2 may be a useful target in gastric cancer.

Effect of hydrodynamics on modelling, monitoring and control of crystallization

Crystallization is a purification method used to obtain crystalline product of a certain crystal size. It is one of the oldest industrial unit processes and commonly used in modern industry due to its good purification capability from rather impure solutions with reasonably low energy consumption. However, the process is extremely challenging to model and control because it involves inhomogeneous mixing and many simultaneous phenomena such as nucleation, crystal growth and agglomeration. All these phenomena are dependent on supersaturation, i.e. the difference between actual liquid phase concentration and solubility. Homogeneous mass and heat transfer in the crystallizer would greatly simplify modelling and control of crystallization processes, such conditions are, however, not the reality, especially in industrial scale processes. Consequently, the hydrodynamics of crystallizers, i.e. the combination of mixing, feed and product removal flows, and recycling of the suspension, needs to be thoroughly investigated. Understanding of hydrodynamics is important in crystallization, especially inlargerscale equipment where uniform flow conditions are difficult to attain. It is also important to understand different size scales of mixing; micro-, meso- and macromixing. Fast processes, like nucleation and chemical reactions, are typically highly dependent on micro- and mesomixing but macromixing, which equalizes the concentrations of all the species within the entire crystallizer, cannot be disregarded. This study investigates the influence of hydrodynamics on crystallization processes. Modelling of crystallizers with the mixed suspension mixed product removal (MSMPR) theory (ideal mixing), computational fluid dynamics (CFD), and a compartmental multiblock model is compared. The importance of proper verification of CFD and multiblock models is demonstrated. In addition, the influence of different hydrodynamic conditions on reactive crystallization process control is studied. Finally, the effect of extreme local supersaturation is studied using power ultrasound to initiate nucleation. The present work shows that mixing and chemical feeding conditions clearly affect induction time and cluster formation, nucleation, growth kinetics, and agglomeration. Consequently, the properties of crystalline end products, e.g. crystal size and crystal habit, can be influenced by management of mixing and feeding conditions. Impurities may have varying impacts on crystallization processes. As an example, manganese ions were shown to replace magnesium ions in the crystal lattice of magnesium sulphate heptahydrate, increasing the crystal growth rate significantly, whereas sodium ions showed no interaction at all. Modelling of continuous crystallization based on MSMPR theory showed that the model is feasible in a small laboratoryscale crystallizer, whereas in larger pilot- and industrial-scale crystallizers hydrodynamic effects should be taken into account. For that reason, CFD and multiblock modelling are shown to be effective tools for modelling crystallization with inhomogeneous mixing. The present work shows also that selection of the measurement point, or points in the case of multiprobe systems, is crucial when process analytical technology (PAT) is used to control larger scale crystallization. The thesis concludes by describing how control of local supersaturation by highly localized ultrasound was successfully applied to induce nucleation and to control polymorphism in reactive crystallization of L-glutamic acid.

Regulation of cell growth, death, and polarization by ERBB4

Regulation of cell growth, death, and polarization by ERBB4 ErbB4 is a member of the epidermal growth factor receptor (EGFR, ErbB) family. The other members are EGFR, ErbB2 and ErbB3. ErbB receptors are important regulators for example in cardiovascular, neural and breast development but control key cellular functions also in many adult tissues. Abnormal ErbB signaling has been shown to be involved in various illnesses such as cancers and heart diseases. Among the ErbBs, ErbB4 has been shown to have unique signaling characteristics. ErbB4 exists in four alternatively spliced isoforms that are expressed in a tissue-specific manner. Two of the isoforms can be cleaved by membrane proteases, resulting in release of soluble intracellular domains (ICD). Once released into the cytosol, the ICD is capable of translocating into the nucleus and participating in regulation of transcription. The functional differences and the tissue-specific expression patterns suggest isoformspecific roles for ErbB4 isoforms. However, the abilities of ErbB4 isoforms to differently regulate cellular functions were discovered only recently and are not well understood. This study aimed to determine the expression patterns of ErbB4 in normal and diseased tissue, and to define whether the cleavable and non-cleavable isoforms could regulate different target genes and therefore, cellular functions. In this study, a comprehensive ErbB4 expression pattern in several normal tissues, various cancers and non-neoplastic diseases was determined. In addition, the data demonstrated that the cleavable and non-cleavable ErbB4 isoforms could regulate different cellular functions and target genes. Finally, this study defined the cellular responses regulated by ErbB4 during kidney development.

RELATIONSHIP BETWEEN CLIMATE VARIABLES, TRUNK GROWTH RATE AND WOOD DENSITY OF Eucalyptus grandis W. Mill ex Maiden TREES

ABSTRACT Climatic conditions stimulates the cambial activity of plants, and cause significant changes in trunk diameter growth and wood characteristics. The objective of this study was to evaluate the influence of climate variables in the diameter growth rate of the stem and the wood density of Eucalyptus grandis trees in different classes of the basal area. A total of 25 Eucalyptus trees at 22 months of age were selected according to the basal area distribution. Dendrometer bands were installed at the height of 1.30 meters (DBH) to monitor the diameter growth every 14 days, for 26 months. After measuring growth, the trees were felled and wood discs were removed at the DBH level to determine the radial density profile through x-ray microdensitometry and then re-scale the average values every 14 days. Climatic variables for the monitoring period were obtained and grouped every 14 days. The effect of the climate variables was determined by maximum and minimum growth periods in assessing trunk growth. These growth periods were related with precipitation, average temperature and relative air humidity. The re-scaled wood density values, calculated using the radial growth of the tree trunks measured accurately with steel dendrometers, enabled the determination of the relationship of small changes in wood density and the effect of the climatic variations and growth rate of eucalyptus tree trunks. A high sensitivity of the wood density to variation in precipitation levels was found.

Towards combining interactive mobile TV and smart spaces : architectures, tools and application development

Technological developments in microprocessors and ICT landscape have made a shift to a new era where computing power is embedded in numerous small distributed objects and devices in our everyday lives. These small computing devices are ne-tuned to perform a particular task and are increasingly reaching our society at every level. For example, home appliances such as programmable washing machines, microwave ovens etc., employ several sensors to improve performance and convenience. Similarly, cars have on-board computers that use information from many di erent sensors to control things such as fuel injectors, spark plug etc., to perform their tasks e ciently. These individual devices make life easy by helping in taking decisions and removing the burden from their users. All these objects and devices obtain some piece of information about the physical environment. Each of these devices is an island with no proper connectivity and information sharing between each other. Sharing of information between these heterogeneous devices could enable a whole new universe of innovative and intelligent applications. The information sharing between the devices is a diffcult task due to the heterogeneity and interoperability of devices. Smart Space vision is to overcome these issues of heterogeneity and interoperability so that the devices can understand each other and utilize services of each other by information sharing. This enables innovative local mashup applications based on shared data between heterogeneous devices. Smart homes are one such example of Smart Spaces which facilitate to bring the health care system to the patient, by intelligent interconnection of resources and their collective behavior, as opposed to bringing the patient into the health system. In addition, the use of mobile handheld devices has risen at a tremendous rate during the last few years and they have become an essential part of everyday life. Mobile phones o er a wide range of different services to their users including text and multimedia messages, Internet, audio, video, email applications and most recently TV services. The interactive TV provides a variety of applications for the viewers. The combination of interactive TV and the Smart Spaces could give innovative applications that are personalized, context-aware, ubiquitous and intelligent by enabling heterogeneous systems to collaborate each other by sharing information between them. There are many challenges in designing the frameworks and application development tools for rapid and easy development of these applications. The research work presented in this thesis addresses these issues. The original publications presented in the second part of this thesis propose architectures and methodologies for interactive and context-aware applications, and tools for the development of these applications. We demonstrated the suitability of our ontology-driven application development tools and rule basedapproach for the development of dynamic, context-aware ubiquitous iTV applications.

Rice seedling and plant development as affected by increasing rates of penoxsulam under controlled environments

Rice is a major staple in many countries. Weed control is one of the factors limiting higher rice yield. ALS (acetolactate synthase)-inhibiting herbicides are desirable weed control herbicides because of their high efficacy, low toxicity to mammalians, and low rates used. An important herbicide characteristic is high selectivity to the crop, since it facilitates fast crop establishment and greater crop advantage over the weeds. The objectives of this work were to study the effects of increasing rates of the ALS-inhibiting herbicide penoxsulam on seed integrity and germination, and seedling and plant development of rice cv. BRS Pelota under controlled laboratory and greenhouse conditions. The results showed that penoxsulam affected rice germination and seedling and plant growth at rates above 54 g a.i. ha-1, and that penoxsulam is safe for rice seedling development at the currently recommended rates.

Vegetative and reproductive development of Costa Rican weedy rice compared with commercial rice (Oryza sativa)

The variability in the chronology of the vegetative and reproductive development of weedy rice complex has been little studied. However, a field trial was established to study the timing of growth stages of sixteen weedy rice morphotypes and five rice varieties of Costa Rica. Weedy rice presented a wide range of variation for all descriptors among and within morphotypes. Weedy rice was taller than the rice varieties during vegetative phase and showed a growth increase of 14-23 cm every two weeks. Six morphotypes emerged earlier than commercial rice varieties, but no differences where found between samples for the time required for starting tillering. Early emergence of weedy rice morphotypes was not associated with early flowering, thus no correlation was detected between the vegetative and reproductive phases. All weedy rice morphotypes reached anthesis and maturity earlier than the rice varieties. Nevertheless, varieties Setesa-9 and CR-5272 overlapped anthesis with eleven morphotypes and variety CR-4338 overlapped flowering with eight weedy rice morphotypes. In contrast, none of the morphotypes overlapped anthesis with varieties CR-1821 and CR-1113. The results obtained showed the competitive capacity of weedy rice and provided valuable information about flowering overlap between weedy rice morphotypes and rice varieties which will be useful in the design of gene flow studies among them.

Primary and secondary development of Cyperus giganteus Vahl rhizome (Cyperaceae)

In Cyperus giganteus, like in other Monocotyledoneae, the protoderm, procambium, fundamental meristem and primary thickening meristem (PTM) are differentiated from the rhizome promeristem. The PTM produces the inner cortical parenchyma, endodermis, pericycle and amphivasal vascular bundles, which are formed by the procambium too. After the primary body differentiates, cellular divisions continue only in the pericycle, and originate an irregular vascular system with vessel elements shorter and more branched than those found in the primary growth. This change of activity in the pericycle defines a secondary growth, where the secondary thickening meristem (STM) is the pericycle itself.

Effect of abscisic acid on the mobilisation of galactomannan and embryo development of Sesbania virgata (Cav.) Pers. (Leguminosae - Faboideae)

Galactomannans (GM) are storage cell wall polysaccharides present in endospermic seeds of legumes. They are thought to be storage polymers, since it has been observed for a few species (among them Sesbania virgata) that they are completely broken down after germination and their products are transferred to the growing embryo. We examined the effect of 10-4 M abscisic acid (ABA) on the degradation of galactomannan in isolated endosperms and intact seeds of S. virgata. We found that after seed germination the initial embryo growth was retarded. Ultrastructural analysis showed that the embryo is completely surrounded by an endosperm which displays very thick galactomannan-containing cell walls. Although an inhibitory effect has been observed on the increase of fresh mass of the embryo, the effect of ABA on the dry mass was weaker and transitory (from 48 to 96 h). Endosperm dry mass and galactomannan degradation were significantly inhibited and the activity of alpha-galactosidase was strongly affected. The addition of ABA before and/or after the start of mobilisation in intact seeds or isolated endosperms, showed that whereas addition before mobilisation did not affect dry mass decrease in intact seeds, it was strongly affected in isolated endosperms. On the other hand, whereas it affected embryo fresh mass increase in intact seeds, but not in isolated embryos, no significant effect was observed on dry mass. These results suggest that ABA affects galactomannan degradation and by doing so, prevents water absorption by the embryo, rather than affect its dry mass. As ABA has been detected in the endosperm of seeds of S. virgata, it is proposed that it probably acts as a modulator of galactomannan mobilisation and consequently synchronises it with early growth of the embryo.

Megaspore germination and initial development of Regnellidium diphyllum Lindman (Pteridophyta, Marsileaceae) sporophytes in the presence of cadmium

Regnellidium diphyllum has its distribution restricted to Southern Brazil and adjoining localities in Uruguay and Argentina. Currently it is on the list of threatened species of Rio Grande do Sul. The conversion of wetlands into agricultural areas or soil contamination by the introduction of waste products and fertilizers may compromise the establishment and survival of this species. Among the pollutants are heavy metals, such as cadmium (Cd). Megaspores were germinated in liquid culture medium, with concentrations 0 (control), 0.39; 0.78; 1.56; 3.12; 6.25; 12.5; 25; 50 and 100 mg L-1 of Cd, starting from a standard solution of Titrisol® at 1000 mg L-1. The increase of Cd in the growth medium to 50 mg L-1 resulted in low germinability (58%), and no germination was observed on 100 mg L-1. In apomictical sporophytes, the growth of primary root and leaf was significantly reduced and no secondary leaf was formed at Cd concentrations of 12.5 and higher than this. The results indicated that R. diphyllum is tolerant to the presence of Cd up to considerably higher concentrations (0.78 mg L-1) than that normally found in unpolluted aquatic ecosystems (0.01 mg L-1), although the sensitivity to higher concentrations might endanger the establishment and permanence of this species in habitats exposed to contamination with this metal.

Do intrauterine growth restriction and overweight at primary school age increase the risk of elevated body mass index in young adults?

Obesity is one of the rising public health problems characterized as a risk factor for many chronic diseases in adulthood. Early life events such as intrauterine growth restriction, as well as life style, are associated with an increased prevalence of this disease. The present study was performed to determine if intrauterine growth restriction interacts with overweight at primary school age to affect body mass index (BMI) in young adults. From June 1, 1978 to May 31, 1979, 6827 singleton liveborns from Ribeirão Preto, São Paulo State, Brazil, corresponding to 98% of all births at the 8 maternity hospitals, were examined and their mothers were interviewed. Samples from the initial cohort were examined again at primary school age (8 to 11 years of age) and at the time of military service (18 years of age). There were 519 male individuals with complete measurements taken in the three surveys. Intrauterine growth-restricted individuals had a BMI 0.68 kg/m² lower than that of individuals who were not restricted (95%CI = -1.34 to -0.03) and overweight at primary school age showed a positive and strong effect on BMI at 18 years of age (coefficient 5.03, 95%CI = 4.27 to 5.79). However, the increase in BMI was much higher - 6.90 kg/m² - when the conscript had been born with intrauterine growth restriction and presented overweight at primary school age (95%CI = 4.55 to 9.26). These findings indicate that the effect of intrauterine growth restrictionon BMI at 18 years of age is modified by later weight gain during school age.

Effect of conditioned medium of mesenchymal stem cells on the in vitro maturation and subsequent development of mouse oocyte

Mesenchymal stem cells (MSCs) secrete a variety of cytokines and growth factors in addition to self-renewal and multiple forms of differentiation. Some of these secreted bioactive factors could improve meiotic maturation in vitro and subsequent embryo developmental potential. The aim of the present study was to determine whether in vitro maturation (IVM) of mouse oocyte with or without cumulus cells could be improved by contact with conditioned medium (CM) of MSCs as well as the efficiency of CM to support follicular growth and oocyte maturation in the ovarian organ of mice cultured on soft agar. The developmental potential of matured oocyte was assessed by blastocyst formation after in vitro fertilization (IVF). Germinal vesicle stage oocytes with or without cumulus cells were subjected to IVM in either CM, Dulbecco's modified Eagle's medium (DMEM), α-minimum essential medium (α-MEM) or human tubal fluid (HTF). Approximately 120 oocytes were studied for each medium. CM produced a higher maturation rate (91.2%) than DMEM (54.7%), α-MEM (63.5%) and HTF (27.1%). Moreover, CM improved embryo development to blastocyst stage significantly more than DMEM and HTF (85 vs 7% and 41.7%, respectively) but there was no significant difference compared with α-MEM (85 vs 80.3%). The behavior of cortical granules of IVM oocytes cultured in CM revealed cytoplasmic maturation. Moreover, CM also supported preantral follicles growth well in organotypic culture on soft agar resulting in the maturation of 60% of them to developmentally competent oocytes. The production of estrogen progressively increased approximately 1-fold every other day during organ culture, while a dramatic 10-fold increase in progesterone was observed 17 h after human chorionic gonadotropin stimulus at the end of culture. Thus, CM is an effective medium for preantral follicle growth, oocyte maturation, and sequential embryo development.

States and Economic Development

Today the Washington Consensus on development lies in tatters. The recent history of the developing world has been unkind to the core claim that a nation that opens its economy and keeps government's role to a minimum invariably experiences rapid economic growth. The evidence against this claim is strong: the developing world as a whole grew faster during the era of state intervention and import substitution (1950-1980) than in the more recent era of structural adjustment (1990-2005); and the recent economic performance of both Latin America and Sub-Saharan Africaregions that truly embraced neoliberalismhas lagged well behind that of many Asian economies, which have instead pursued judicial and unorthodox combinations of state intervention and economic openness. As scholars and policy makers reconstruct alternatives to the Washington Consensus on development, it is important to underline that prudent and effective state intervention and selective integration with the global economy have been responsible for development success in the past; they are also likely to remain the recipes for upward mobility in the global economy in the future."

Capacity and transformational development within the 2005 Canada Summer Games host society

Although capacity has been used in recent federal government accords and policies related to the voluntary and amateur sport sectors, there is little consensus over the meaning of the term. Consequently, the purpose of this qualitative case study was to explore the concept of organizational capacity within a temporary voluntary sport organization. Specifically, the nature of organizational capacity was examined within the case of the Volunteers Division of the 2005 Canada Summer Games (CSG) Host Society. Data were collected from executive planning and middle management CSG volunteers through the use of a variety of methods: verbal journals, interviews, observations, documents and a focus group. Findings indicated several challenges associated with the volunteer management model utilized by the host society, varying levels of importance among six elements of capacity, and key aspects of the relationship between organizational capacity and transformational development. Implications focused upon the importance of highlighting individuals rather than the organizational as a whole in order to build capacity, and utilizing a brain or hybrid brain-machine organizational form to enhance capacity. Recommendations are provided for both the Canada Games Council and Canada Games host societies.

Effect of a high fat maternal diet on body composition and bone development in male offspring

Direct high fat (HF) feeding has adverse effects on body composition and bone development in rodents. However, it is unclear whether maternal HF feeding has similar effects in male rat offspring. The objectives of this thesis were to determine if maternal HF feeding altered body composition, plasma hormones, bone development, and bone fatty acid composition in male offspring at weaning and 3 months of age. Maternal HF feeding increased bone mass and altered femur fatty acid composition at weaning, without differences in fat mass, lean mass, plasma hormones, or bone mass (femur or lumbar vertebrae). However, early differences did not persist at 3 months of age or contribute to lower bone strength – following consumption of a control diet post-weaning. These findings suggest that maternal HF feeding can alter body composition and bone development in weanling male offspring, without long-lasting effects if a healthy control diet is consumed post-weaning.