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The investigation was concerned with characterizing effects of this tumor line on lipid metabolism in DBA/1J mice and serum protein levels and cellular changes in DBA/1J and Swiss white mice. Total lipids, lipid phosphorus, neutral lipids, and changes in fatty acids were determined in liver, spleen, skin and tumor of DBA/1J mice bearing the lymphosarcoma at various days after injection of tumor cells.

Hereditary Spherocytosis (HS) is a common inherited haemolytic anaemia with variable clinical expression. Fifty subjects with HS from KwaZulu-Natal were studied with the aim of providing further information on the protein abnormalities of the red blood cell (RBC) membrane and their relationship with clinical presentations. Haematological and biochemical tests were performed by routine procedures. Mean Corpuscular Haemoglobin Concentration ( MCHC) in the HS group was 35.1g /dl. This was significantly higher than in normal control subjects (33.6g /dl) (p value < 0.001); indicating its usefulness for the screening of HS. Mean Red Cell Distribution Width (RDW) was also significantly higher in subjects with HS (p<0.001); thus providing an additional screening tool. Erythrocyte membrane proteins from 21 subjects were analysed by SDS - polyacrylamide gel electrophoresis (SDS-PAGE) using the Laemmli and Fairbanks methods. The most common abnormality was a deficiency of band 3 (10 subjects), followed by a combined spectrin and ankyrin deficiency in five subjects. One subject had increased band 6 and in five cases no abnormality was detected. A decreased ratio of protein 4.1a / 4.1b on the Laemmli SDS PAGE correlated with an increased reticulocyte count. The degree of haemolysis and clinical findings did not correlate with the type of red cell membrane protein defect. In this study red cell membrane analysis did not contribute further to the initial laboratory diagnosis. In addition it did not influence clinical management. The presence of red cell membrane abnormalities, either single or multiple, did not correlate with disease severity. Red cell membrane analysis, however, will play an important role for future management such as gene therapy. Red cell membrane analysis is also useful as a research tool to determine the underlying molecular defect and to assess racial or ethnic differences. It is also of value as a differential diagnostic tool in cases where the clinical and laboratory findings are not conclusive for HS. / Thesis (M.Med.Sci)-University of KwaZulu-Natal, Durban, 2006.

This investigation was concerned with characterizing a tumor line induced and maintained in this laboratory. Various chemical assays, cell counts, and electron microscopy were the methods employed to characterize the blood of mice bearing the tumor at days 3, 6, 9, and 12 after injection of the 1.2 x 10^8 tumor cells.

Regulation of hematopoiesis through the finite control of hematopoietic stem cell (HSC) quiescence and proliferation is critical to the health of the organism since disturbances in blood production can lead to clinical malignancies such as anemia or leukemia. Therefore, elucidating the processes that govern HSCs is vital to advance our understanding of hematological diseases. Interestingly, HSCs can be regulated through a variety of ways. Extrinsic cues from the niche provide signals that govern HSC quiescence, proliferation, self-renewal, and differentiation. These external signals are converted to internal messages through the use of signal transduction pathways that relay information from the cytoplasm to the nucleus. While many pathways contribute to HSC regulation, the PI3K/AKT/mTOR-pathway is especially pertinent because it has been implicated in cell survival, metabolism, proliferation, and death. Many groups have identified key players within PI3K/AKT/mTOR-signaling that regulate HSC quiescence; however, these studies are hindered by the redundancy of multiple isoforms and compensatory signaling mechanisms by other members within the pathway. PDK1 is considered to be a master regulator of PI3K-signaling because it is directly activated by PI3Ks and can govern activity of a variety of substrates within the PI3K-pathway. Because of this, it is an excellent candidate to fully delineate how PDK1-mediated PI3K-signaling functions to maintain HSC quiescence. In the current study, conditional deletion of PDK1 in HSCs was achieved through the use of a hematopoietic specific Vav-Cre line. The loss of PDK1 in HSCs led to reduced numbers and an inability to provide radioprotection in primary BMTs. Furthermore, PDK1-deficient HSCs exhibit impaired quiescence and increased cycling. Strikingly, PDK1-mutant HSCs have markedly high levels of reactive oxygen species (ROS), which led to increased proliferation, DNA damage, and apoptosis in progenitor cells. Administration of a ROS scavenger, N-acetyl cysteine (NAC) partially rescued the increased proliferation and differentiation of phenotype Pdk1Hem-KO cells both in vitro and in vivo, suggesting that abnormal HSC activity in PDK1-deficient cells was in part due to increased ROS. Furthermore, mechanistic studies identified a remarkable decrease in the levels of nuclear HIF1α in HSCs. Immunoblots and phosflow studies demonstrated reduced activation of p-p70S6K, a well defined positive regulator, and increased GSK3β, a key negative regulator of the HIF1α protein. These data suggested that ROS levels are unrestrained since HIF1α is not present in Pdk1Hem-KO HSCs to activate transcription of genes that moderate oxidative stress. In addition, Pdk1Hem-KO HSCs also show increased levels of Hif3α and IPAS mRNA, which are believed to inhibit the transcriptional activation of HIF1α. In essence, the results from these experiments are the first to implicate PDK1 as a critical regulator of HSC quiescence and as a moderator of PI3K-signaling to alleviate oxidative stress. In addition, this study is the first to suggest that HIF3α and IPAS may play a role in HSCs.

This dissertation examines risk factors and outcomes associated with bloodstream infection (BSI). In Chapter One, the problems of BSI are introduced and their significance described. In Chapter Two, the results of a systematic review and meta-analysis synthesizing the prevalence of one of the most rapidly emerging causes of BSI among nursing home residents, multidrug resistant-gram negative bacteria are described. In Chapter Three, a retrospective cohort study identifying the prevalence and risk factors for BSI present on hospital admission (POA) is reported, including an assessment of antimicrobial resistance in isolates causing BSI-POA by admission source (i.e. private homes, other hospitals and skilled nursing facilities). In Chapter Four, a retrospective cohort study explaining risks for hospital-associated infections (HAIs) among the BSI-POA cohort is described. Length of stay and mortality among patients with a BSI-POA who develop HAI and those who do not are reported. Finally, in Chapter Five, findings of the previous chapters are synthesized and the conclusion is provided including strengths, limitations and implications for policy and practice.

Background: Bernard Soulier Syndrome (BSS) is a rare, autosomal recessive inheritance disorder of platelet function. Estimated to affect one per one million, there are currently only 200 cases reported worldwide presenting more commonly in families with parental consanguinity. This syndrome occurs when there is a genetic defect in the subunits (GPIb-alpha, GPIB-beta, and GP9) that form the GPIb-IX-V complex. The result is inadequate binding to von Willebrand factor. The clotting cascade is, therefore, unable to begin, causing symptoms of excessive and prolonged bleeding. Objectives: We report a case with multiple episodes of exaggerated bleeding and easy bruising. Methods: We analyzed complete blood count, coagulation studies, platelet aggregation assays, platelet glycoprotein expression by flow cytometry, as well as screened both patient and parents for relevant genes responsible for BSS. Results: 14-month-old Caucasian male born at 38w3d gestational age, non-consanguineous parents with multiple episodes of exaggerated bleeding and easy bruising from minor injuries. His symptoms started early in life with excessive bleeding after circumcision. No history of intramuscular, joint, or intracranial bleeding. Complete blood counts showed macrothrombocytopenia (98 X109 /L MPV 12.3 fl) no leukocyte inclusion bodies on peripheral smear. Coagulation tests (prothrombin time, activated partial thromboplastin time, vWF antigen, and vW-Ristocetin cofactor activity, platelet function assay) were normal. Platelet glycoprotein expression by flow cytometry revealed significantly reduced binding of monoclonal antibodies to platelet GPIb and normal GPIIb/IIIa. Comprehensive platelet disorder panel revealed two clinically significant variants missense mutations in the GP9 gene (P.Cys135 Tyr and P.Asn61Ser) These variants were on opposite alleles and results were consistent with the diagnosis of Bernard Soulier syndrome (BSS). The mother reported heavy menstrual cycles, the father had no significant bleeding symptoms, and both parents had normal platelet counts. Target genetic testing identified these two distinct missense mutations from both Mother and Father of the child. Conclusion: The two rare variants occurring on the gene for GPIX (GP9) increase the number of known genetic defects associated with the manifestation of Bernard Soulier Syndrome.

Este estudo teve o propósito de validar os diagnósticos de enfermagem da NANDA (2001) \"Disfunção Sexual\" e \"Padrões de Sexualidade Ineficazes\" e de proporcionar uma avaliação objetiva da Sexualidade Humana. A abordagem metodológica de validação adotada foi a de Hoskins (1989): análise de conceito, validação por especialistas e validação clínica, denominada de verificação da incidência das evidências clínicas. A partir da reconstrução dos dois diagnósticos estudados e construção das definições operacionais de suas características definidoras, estes foram validados por 32 enfermeiros considerados peritos nestes diagnósticos, a partir da pontuação recomendada por Fehring (1994). Os resultados demonstraram que no diagnóstico \"Disfunção Sexual\" a definição está adequada ao título, mas merece que se especifique as fases da resposta sexual humana. Quanto às 10 características definidoras propostas, 07 receberam os maiores escores: limitações percebidas impostas pela doença e/ou terapêutica; limitações reais impostas pela doença e/ou terapêutica; alteração no alcance da satisfação sexual; busca de confirmação da qualidade de ser desejável; incapacidade de alcançar a satisfação desejada; percepção de alteração na excitação sexual e percepção de escasso desejo sexual. Ressalta-se que essas duas últimas características foram encontradas na literatura e acrescidas neste diagnóstico. Referente ao diagnóstico \"Padrões de Sexualidade Ineficazes\", a adequação da definição ao título necessitou de ser comprovada pela literatura, pois as sugestões dos enfermeiros peritos que discordaram foram heterogêneas. Quanto às características definidoras, os maiores escores foram atribuídos a dificuldades ou limitações relatadas nos comportamentos ou atividades sexuais, mudanças relatadas nos comportamentos ou atividades sexuais, alterações no desempenho do papel sexual percebido e alteração no relacionamento com pessoas significativas, sendo estas duas últimas foram deslocadas do diagnóstico \"Disfunção Sexual\". Todas as características definidoras do diagnóstico \"Disfunção Sexual\" que receberam escores acima de 0,80 na Validação por Especialistas tiveram incidências acima de 55% no contexto clínico dos pacientes portadores de doenças onco-hematológicas. Porém, referente ao diagnóstico \"Padrões de Sexualidade Ineficazes\", as maiores incidências ocorreram nas características definidoras: dificuldades ou limitações relatadas nos comportamentos ou atividades sexuais e mudanças relatadas nos comportamentos ou atividades sexuais, que constam na NANDA (2001). Conclui-se que os pacientes portadores de neoplasias hematológicas apresentaram comprometimento de uma ou mais fases da resposta sexual humana e mostraram-se preocupados com a influência da doença e do tratamento no seu comportamento sexual, principalmente, no que se refere ao medo de se adquirir uma infecção, resultando na diminuição ou interrupção da sua atividade sexual, após descobrir a doença / This study aimed to validate the NANDA (2001) nursing diagnoses \"Sexual Dysfunction\" and \"Ineffective Sexuality Patterns\" and offer an objective evaluation of Human Sexuality. Our methodological approach of validation was based on Hoskins (1989): concept analysis, expert validation and clinical validation, called verification of clinical evidence incidence. Starting from the reconstruction of the two diagnoses under analysis and the construction of the operational definitions of their defining characteristics, validation was realized by 32 nurses, who are considered experts on these diagnoses, according to the punctuation recommended by Fehring (1994). Results demonstrate that, with respect to the \"Sexual Dysfunction\" diagnosis, the definition is adequate for the title, but human sexual response phases need to be specified. 07 out of 10 defining characteristics obtained the highest score: perceived limitations imposed by disease and/or therapy; actual limitations imposed by disease and/or therapy; alteration in achieving sexual satisfaction; seeking confirmation about being desirable; incapacity to achieve desired satisfaction; perception of altered sexual excitement and perception of scarce sexual desire. We highlight that the last two characteristics were found in literature and added to this diagnosis. The adequacy of the \"Ineffective Sexuality Patterns\" definition to the title had to be proved by means of literature, since the experts nurses displayed heterogeneous suggestions in this respect. The defining characteristics with the highest scores were: reported difficulties or limitations in sexual behaviors or activities; reported changes in sexual behaviors or activities; alterations in achieving perceived sex role and alteration in relationship with significant other. The last two were transferred from the \"Sexual Dysfunction\" diagnosis. All defining characteristics for the \"Sexual Dysfunction\" diagnosis with scores superior to 0.80 in Experts Validation disclosed incidence rates superior to 55% in the clinical context of blood cancer patients. However, with respect to the \"Ineffective Sexuality Patterns\" diagnosis, the highest incidence rates occurred for the following defining characteristics: reported difficulties or limitations in sexual behaviors or activities and reported changes in sexual behaviors or activities, which are listed in NANDA. We conclude that the human sexual response of blood cancer patients was affected in one or more phases. Patients revealed to be worried about the influence of the disease and treatment on their sexual behavior, mainly with respect to the fear of getting an infection, resulting in decreased or interrupted sexual activity after the discovery of the disease

"May 2002" Includes bibliographical references (leaves 229-251) Describes the investigation of the alteration of ABH antigen expression on the surface of red blood cells in patients with haematological malignancies.

[Truncated abstract] Iron is essential to most biological systems. Under pathological conditions affecting the iron metabolic pathway, iron can be deposited in the tissue in various forms. The work presented in this thesis has exploited the relationship between the magnetic and the chemical properties of tissue iron deposits to further understanding of two major pathologies, the haemoglobinopathies termed thalassaemias and the malaria parasite Plasmodium falciparum, both amongst the most common health concerns in tropical countries. The iron-specific magnetic susceptibilities ¿Fe for spleen tissue samples from 7 transfusion dependent ß-thalassaemia (ß-thal) patients and 11 non-transfusion dependent ß-thalassaemia/Haemoglobin E (ß/E) patients were measured at 37°C. Both groups of patients were iron loaded with no significant difference in the distribution of spleen iron concentrations between the two groups. There was a significant difference between the mean ¿Fe of the spleen tissue from each group. The ß/E patients had a higher mean (± standard deviation) spleen ¿Fe (1.55 ± 0.23 × 10-6 m3.kgFe -1) than the ß-thal patients (1.16 ± 0.25 × 10-6 m3.kgFe -1). Correlations were observed between ¿Fe of the spleen tissue and the fraction of magnetic hyperfine split sextet in the 57Fe Mössbauer spectra of the tissues at 78 K (Spearman rank order correlation ¿ = -0.54, p = 0.03) and between ¿Fe of the spleen tissue and the fraction of doublet in the spectra at 5 K (¿ = 0.58, p = 0.02) indicating that ¿Fe of the spleen tissue is related to the chemical speciation of the iron 2 deposits in the tissue. The biological variability of the iron-specific magnetic susceptibility of the tissue iron examined would contribute a random uncertainty of 19% to magnetic susceptibility based non-invasive measurements of tissue iron concentration. ... Magnetic susceptibility measurements were also performed on malaria parasitised red blood cells. In vitro cultures of P. falciparum were magnetically enriched up to 61-fold using high field gradient magnetic separation columns, and the magnetic susceptibility of cell contents was directly measured. Forms of haem iron were quantified spectroscopically. Further fractionations were performed such that, by controlling the fluid velocity through the column, cells with more than a critical amount of paramagnetic 3 iron were preferentially extracted. A chloroquine-sensitive (CQS) laboratory strain of parasites converted approximately 60% of host cell haem iron to haemozoin and this product was the primary source of the increase in cell magnetic susceptibility. The volumetric magnetic susceptibility of the magnetically enriched cells was found to be 0.15 ± 0.03 × 10-7 relative to the suspension medium, accounting for the enrichment of mature parasites. Comparisons of fractionation samples of two pairs of CQS and chloroquine resistant (CQR) strains showed enrichment of mature parasites was significantly greater in the CQS than the CQR strains. The results suggest the possibility of using magnetic separation columns in identifying CQR strains of P. falciparum, potentially in a diagnostic or research setting. The study also underlines the need to identify and quantify the forms of iron in CQR and CQS parasite strains as the fate of haem iron will have implications in understanding the mechanisms of chloroquine resistance.