The research presented in this dissertation harnesses two secondary data sources, administrative databases of patient-level healthcare data and adverse event (AE) data reported in randomized clinical trials (RCTs), to assess the relationship between intravenous immune globulin (IVIg) and the risk of clinically serious thromboembolic adverse events (TEEs). Since 2013, IVIg products have carried a boxed warning concerning TEE risk, a determination supported by numerous case reports, a large claims-based risk assessment, and laboratory evaluations of the thrombogenecity of IVIg products. Questions remain concerning the magnitude of the risk overall and across subgroups of IVIg users. Taken together, our results are compatible with the conclusion that the absolute risk of TEE following IVIg use is likely to be low overall. While these results are reassuring, a clinically meaningful elevation in risk cannot be ruled out in certain patient sub-groups, such as older adults and others with a high baseline risk of TEE. A limitation of our research is that differences in TEE risk across products could not be evaluated with sufficient statistical power.

The cereal seed plays a crucial role in society – both in the “food as medicine” paradigm, but also in food security. It is the starch and proteins present in the seed that lend it importance in these dissimilar anthropomorphic activities. This thesis investigation first characterized the post-translational processing of the potential diabetogen, wheat globulin-3. Globulin-3-like peptides were observed primarily in the embryo. These peptides varied significantly in their molecular masses and isoelectric points, as determined by two dimensional electrophoresis and immunoblotting. Five major polypeptide spots were sequenced by mass spectrometry, allowing for the development of a model of the post-translational events contributing to the globulin-3 processing profile. Three separate investigations of starch granules from different cereal species were performed. In the first series of experiments, pathogen-susceptible maize kernels were injected with either conidia of the fungal pathogen Fusarium graminearum or sterile water controls. Proteins in the desiccated fungal remnants on the surface of the kernels as well as in the endosperm and embryo tissues of the control and infected kernels were isolated and these proteomes were sequenced using tandem mass spectrometry. Approximately 250 maize proteins were identified. These proteins were classified into functional categories. There was an increased representation of defense proteins in the both the embryo and endosperm tissues of infected maize samples. The proteome of the fungal remnants was composed of 18 proteins. Several of these proteins were categorized as being involved in the metabolism of plant-sourced molecules, or in stress response. The second series of experiments detail the investigation of commercially prepared rice and maize starches using tandem mass spectrometry. The majority of identified proteins, in both rice and maize samples, were involved in either carbohydrate metabolism or storage. Markers for seed maturity and for starch mobilization were also documented. Finally, the third series of experiments investigated the non-host proteomes present in commercially-prepared starches. Non-host proteins from a variety of species, including Homarus americanus were found in the starch samples. This documentation of H. americanus proteins in these starch samples may have food safety implications with regards to shellfish allergies.

The cereal seed plays a crucial role in society – both in the “food as medicine” paradigm, but also in food security. It is the starch and proteins present in the seed that lend it importance in these dissimilar anthropomorphic activities. This thesis investigation first characterized the post-translational processing of the potential diabetogen, wheat globulin-3. Globulin-3-like peptides were observed primarily in the embryo. These peptides varied significantly in their molecular masses and isoelectric points, as determined by two dimensional electrophoresis and immunoblotting. Five major polypeptide spots were sequenced by mass spectrometry, allowing for the development of a model of the post-translational events contributing to the globulin-3 processing profile. Three separate investigations of starch granules from different cereal species were performed. In the first series of experiments, pathogen-susceptible maize kernels were injected with either conidia of the fungal pathogen Fusarium graminearum or sterile water controls. Proteins in the desiccated fungal remnants on the surface of the kernels as well as in the endosperm and embryo tissues of the control and infected kernels were isolated and these proteomes were sequenced using tandem mass spectrometry. Approximately 250 maize proteins were identified. These proteins were classified into functional categories. There was an increased representation of defense proteins in the both the embryo and endosperm tissues of infected maize samples. The proteome of the fungal remnants was composed of 18 proteins. Several of these proteins were categorized as being involved in the metabolism of plant-sourced molecules, or in stress response. The second series of experiments detail the investigation of commercially prepared rice and maize starches using tandem mass spectrometry. The majority of identified proteins, in both rice and maize samples, were involved in either carbohydrate metabolism or storage. Markers for seed maturity and for starch mobilization were also documented. Finally, the third series of experiments investigated the non-host proteomes present in commercially-prepared starches. Non-host proteins from a variety of species, including Homarus americanus were found in the starch samples. This documentation of H. americanus proteins in these starch samples may have food safety implications with regards to shellfish allergies.

Plasma protein shave many important functions in the organism. Gamma globulins are carriers of immunoglobulins which play animportant role in the immune response. Their contentis primarily given by the health burden of the organism. The aimofthe study was to determine the individual protein fractions in the blood plasma of ewes and lambs, comparemutual relations between total plasma proteins and thein variol factions and assess the concentrations of individual plasma proteins, mainly globulin, in connection with the aktivity of the thyroidgland, physiological state, and increments. The sampling was carried out in the spring (25.3.) and the autumn (14.10) 2013. The individual protein fractions were determined from the serum by the means of electrophoresis. The results show that the concentration of proteins in the blood plasma of bothewes and ewe lambs and ram lambs was high due to haemoconcentration, heat stress during sampling, grazing young green forage, comprising a large number of protein aceous substances, and increasing demands on energy for milk production, especially in the early stagesoflactation. Theconcentrationofproteins in theblood plasma oflambspertains to thegrowthproduction. It establishes a correlation between the concentration of plasma proteins and the aktivity of the thyroidgland. It also confirmes higher concentrations of plasma proteins of lambs correlating to thein higher daily gain. Due to the high concentration of total protein in the plasma, the level of its fractions was high as well. After the conversion to a percentage, the level of - globulins in blood plasma in all categories was within normal limits in the range of 14-27%. The concentration of - globulins in blood plasma increased in relation to the parasitological findings coccidia of the genus Eimeria and gastrointestinal nematodes. Relations between the kontent of plasma proteins in the blood of the lambs and ewe sobserved were in most casespositive. There was a strong dependence between the total protein and globulins. Even among other fractions of plasma proteins and globulins correlation coefficient was almou always positive.

A transferência de imunidade passiva colostral é muito importante tanto para a saúde do neonato quanto para o seu desempenho. Com o objetivo de estudar a relação entre a eficiência ou não no estabelecimento da imunidade passiva colostral em neonatos e a suas interrelações com aspectos de desempenho e produção destes animais foram colhidas amostras de sangue de 354 bezerras com 30 horas de vida (24 a 36 h) em 10 propriedades localizadas no centro leste do Estado do Paraná, Brasil. Determinações bioquímicas do soro das amostras foram realizadas para avaliar a qualidade de transferência de imunidade passiva. A proteína sérica total foi determinada pelo método do Biureto e a albumina pelo método do verde de bromocresol para o cálculo das taxas de globulinas obtidas pela diferença entre os teores séricos de proteína e albumina. A fração gamaglobulina foi determinada por eletroforese. Uma análise multivariada, incluindo os teores séricos de proteína total, globulinas e gamaglobulinas, foi utilizada para determinar três grupos conforme à qualidade da transferência de imunidade passiva colostral apresentada (Proc cluster, SAS), a saber: baixa, moderada e alta. Durante o estudo, os produtores anotaram as informações referentes a práticas de manejo adotado na atenção a vaca (parturiente) e ao bezerro, incluindo os dados que poderiam ter influência sobre a transferência de imunidade passiva colostral. Os dados das bezerras, enquanto neonatas e depois quando adultas foram colhidos para a determinação da influência a curto e a longo prazo da falha da transferência de imunidade passiva (FTIP) colostral. Observou-se que os dados de escore de condição corporal da mãe da bezerra, da quantidade de colostro ingerida na primeira mamada e o momento em que ela foi realizada, da morbidade e da mortalidade das bezerras estavam relacionados com a FTIP (P<0,05). Os pesos ao nascer e ao primeiro mês de vida, assim como a freqüência de bezerras analisadas que pariram na propriedade não estavam relacionados diretamente com a qualidade da transferência de imunidade passiva. Fatores como a distocia, idade ao primeiro serviço e a produção média diária de leite não diferiram estatisticamente entre os grupos de baixa, moderada e alta transferência de imunidade passiva (P>0,05). Mas quando se correlacionou por regressão os dados de produção de leite das vacas que quando bezerras apresentaram teores de gamaglobulinas menores que 1,6 g/dL obteve-se valores de r2=0,47 (P=0,0005). Com os resultados desta pesquisa pode-se afirmar que práticas muito simples de manejo como fornecer pelo menos 2 L de colostro até 2 horas após o nascimento e a vaca parir numa boa condição corporal (ECC = 3 ou 3,5) podem prevenir a falha na transferência de imunidade passiva. A FTIP tem como conseqüências: maiores taxas de morbidade e mortalidade, primeiro parto mais tardio e diminuição do número de novilhas de reposição, podendo ainda estar correlacionada a menores produções leiteiras. / Adequate passive transfer of maternal immunoglobulin is important for optimal health and performance in newborn calves. Blood samples were collected from 354 dairy calves, ranging from 24 to 36 hours of age, between July 2005 and May 2006 on 10 farms in the middle-eastern region of the state of Parana, Brazil. The objective was to study the relationship and effectiveness of the transfer of colostral passive immunity and its contributing factors as related to the development and production of animals. For each sample collected, total serum protein was determined by the biuret method and albumin by bromocresol green method, and the difference was used to evaluate the globulins. Electrophoresis was used to determine the -globulin fraction of the sample. A multivariable analysis, including total serum protein, globulins and gamma globulin, was used to create 3 groups to classify the quality of the transfer of colostral passive immunity (cluster procedure, SAS): failure or inadequate group, marginal group and adequate group. During the study, breeders were asked to provide information on calf and pre-partum cow management practices, including details on colostrum feeding. Data from the calves while newborn and as heifers was gathered to determine the long and short term effect of the failure of passive immunity transfer (FPIT). Body condition score of the mother at calving, quantity of colostrum ingested, timing of ingestion, morbidity and mortality of calves and age at calving time were related to FPIT (P<0,05). The weight of the calves after birth and at one month of age and the frequency of calves that became cows in the farm were not directly related to failure. Dystocia, age at first service in days, and milk production did not differ statistically (P>0,05). However, when a regression was performed based on data of milk production from calves that had serum gamma globulins levels below 1,6g/dL, a correlation was identified (r2=0.47; P=0.0005). Basic management practices can prevent failure of passive immunity transfer by feeding calves 2 L of colostrum within 2 hours of life and ensuring that the cow calves with a good body condition score (BCS = 3 or 3.5). FPIT is responsible for higher morbidity and mortality rates, a delay in first parturition, a decrease in the number of replacement heifers and it can also be responsible for less milk production.

Growth and deposition of amyloid fibrils, polymers of proteins with a cross beta-sheet structure, are associated with a significant number of human pathologies including Alzheimer’s disease, Parkinson’s disease, prion diseases, type II diabetes, and senile systematic or dialysis-related amyloidoses. The broader objective of my research is to identify the basic mechanisms regulating nucleation and growth of amyloid fibrils. There is increasing evidence that amyloid formation may proceed along at least two distinct assembly pathways for the formation of rigid fibrils. One pathway involves the nucleated polymerization of the characteristic rigid fibrils from partially denatured monomers and the other proceeds via the growth of globular oligomers and their associated curvilinear fibrils (also known as protofibrils) which, in ways yet to be determined, transform into late-stage rigid fibrils. These oligomeric intermediates of fibril assembly, in particular, have been implicated as the predominant aggregate species causing cellular toxicity in amyloid diseases. Yet, amyloid oligomers and curvilinear fibrils are considered transient, metastable aggregates. This raises the question whether and how such transient aggregate species can be responsible for most of the cell/tissue toxicity? In this dissertation, I report on my investigation of several basic questions related to the mechanisms of amyloid formation. Using the model amyloid hen egg-white lysozyme, I participated in research to characterize the distinct kinetics of amyloid formation along distinct assembly pathways, to determine the morphological features of the various aggregate species emerging along either pathway, and to investigate the structural evolution of the monomers from their native state to the amyloid cross- sheet structure (chapter 3). Chapters 4-6 represent the core of my dissertation work. There I investigated whether amyloid aggregates from three different amyloid proteins, formed under denaturing condition, could undergo prion-like proliferation upon return to physiological solution conditions. I was also intimately involved in a project on the conditions inducing amyloid spherulites formation by polyglutamic acid and the mechanisms resulting in the formation of this often-overlooked amyloid aggregate structure (chapter 7). In the appendix I provide a short summary of the various experimental techniques I have used in the above experiments.

The cereal seed plays a crucial role in society – both in the “food as medicine” paradigm, but also in food security. It is the starch and proteins present in the seed that lend it importance in these dissimilar anthropomorphic activities. This thesis investigation first characterized the post-translational processing of the potential diabetogen, wheat globulin-3. Globulin-3-like peptides were observed primarily in the embryo. These peptides varied significantly in their molecular masses and isoelectric points, as determined by two dimensional electrophoresis and immunoblotting. Five major polypeptide spots were sequenced by mass spectrometry, allowing for the development of a model of the post-translational events contributing to the globulin-3 processing profile. Three separate investigations of starch granules from different cereal species were performed. In the first series of experiments, pathogen-susceptible maize kernels were injected with either conidia of the fungal pathogen Fusarium graminearum or sterile water controls. Proteins in the desiccated fungal remnants on the surface of the kernels as well as in the endosperm and embryo tissues of the control and infected kernels were isolated and these proteomes were sequenced using tandem mass spectrometry. Approximately 250 maize proteins were identified. These proteins were classified into functional categories. There was an increased representation of defense proteins in the both the embryo and endosperm tissues of infected maize samples. The proteome of the fungal remnants was composed of 18 proteins. Several of these proteins were categorized as being involved in the metabolism of plant-sourced molecules, or in stress response. The second series of experiments detail the investigation of commercially prepared rice and maize starches using tandem mass spectrometry. The majority of identified proteins, in both rice and maize samples, were involved in either carbohydrate metabolism or storage. Markers for seed maturity and for starch mobilization were also documented. Finally, the third series of experiments investigated the non-host proteomes present in commercially-prepared starches. Non-host proteins from a variety of species, including Homarus americanus were found in the starch samples. This documentation of H. americanus proteins in these starch samples may have food safety implications with regards to shellfish allergies.

Following rabies virus (RABV) exposure, a combination of thorough wound washing, multiple-dose vaccine administration and the local infiltration of rabies immune globulin (RIG) are essential components of modern post-exposure prophylaxis (PEP). Although modern cell-culture-based rabies vaccines are increasingly used in many countries, RIG is much less available. The prohibitive cost of polyclonal serum RIG products has prompted a search for alternatives and design of anti-RABV monoclonal antibodies (MAbs) that can be manufactured on a large scale with a consistent potency and lower production costs. Robust in vitro neutralization activity has been demonstrated for the CL184 MAb cocktail, a 1:1 protein mixture of two human anti-RABV MAbs (CR57/CR4098), against a large panel of RABV isolates. In this study, we used a hamster model to evaluate the efficacy of experimental PEP against a lethal challenge. Various doses of CL184 and commercial rabies vaccine were assessed for the ability to protect against lethal infection with representatives of four distinct bat RABV lineages of public health relevance: silver-haired bat (Ln RABV); western canyon bat (Ph RABV); big brown bat (Ef-w1 RABV) and Mexican free-tailed bat RABV (Tb RABV). 42–100% of animals survived bat RABV infection when CL184 (in combination with the vaccine) was administered. A dose-response relationship was observed with decreasing doses of CL184 resulting in increasing mortality. Importantly, CL184 was highly effective in neutralizing and clearing Ph RABV in vivo, even though CR4098 does not neutralize this virus in vitro. By comparison, 19–95% survivorship was observed if human RIG (20 IU/kg) and vaccine were used following challenge with different bat viruses. Based on our results, CL184 represents an efficacious alternative for RIG. Both large-scale and lower cost production could ensure better availability and affordability of this critical life-saving biologic in rabies enzootic countries and as such, significantly contribute to the reduction of human rabies deaths globally.

No description available.

Binding parameters of soluble Type I and Type II receptors were assessed in hippocampus of adult, adrenalectomized, male and female rats. No sex differences in the number of either Type I or Type II receptors could be demonstrated between gonadally intact animals. When females treated with 17β-estradiol benzoate (10 μg/day) were compared with males, a statistically significant reduction in Type II receptors was observed in the females; progesterone produced no further decrease in receptor numbers. The amount of tissue-associated corticosteroid-binding globulin in gonadally intact animals (perfused with dextran-saline) was twice as great in females as males. Sex-dependent differences in these gonadally intact rats were found in the affinity, measured as the dissociation constant (Kd), of both the Type I and Type II receptors. For both receptors, affinity in cytosols from females was reduced. The difference for the Type II receptor was slight, but the Kd value of the type I receptor was several-fold higher in females. The difference in affinity was evident with both natural and synthetic steroid ligands. There appears to be little, if any, difference in affinity between the hippocampal Type I and the Type II receptors in females. This suggests that the occupancy of Type I receptors in females is substantially less than that of males at low circulating concentrations of corticosteroids.