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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Evaluation of fluorescence immunophenotyping and interphase cytogenetics as a tool for investigation of neoplasm (FICTION) in aclinical laboratory and its application on clinical blood and marrowspecimens

鄺錦強, Kong, Kam-keung. January 2009 (has links)
published_or_final_version / Pathology / Master / Master of Medical Sciences
2

Evaluation of fluorescence immunophenotyping and interphase cytogenetics as a tool for investigation of neoplasm (FICTION) in a clinical laboratory and its application on clinical blood and marrow specimens

Kong, Kam-keung. January 2009 (has links)
Thesis (M. Med. Sc.)--University of Hong Kong, 2009. / Includes bibliographical references (p. 44-48).
3

Flow cytometry in diagnostic haematopathology

Glencross, Deborah Kim January 1992 (has links)
A Research Report submitted to the Faculty of Medicine, Department of Haematology, University of the Witwatersrand, Johannesburg for the Degree of Master of Medicine (in the branch Haematology). / WHSLYP2016
4

Immunophenotypic classification of canine malignant lymphoma in formalin-fixed, paraffin wax-embedded specimens using CD3 and CD79a cell markers

Pearson, Joyce. January 2009 (has links)
Thesis (MMedVet (Pathology, Veterinary Science))--University of Pretoria, 1999. / Also available in print format.
5

Immunophenotyping of Lymphoid Cells in Autism

Yonk, L. Jeanne 01 May 1991 (has links)
Research into the cause of autism continues without any clear-cut answers. However, recent studies suggest that abnormalities of the immune system are associated with this disorder and autism results from failure of the immune system to regulate itself. Proper immune regulation requires that the host have the appropriate number and percentage of each population of lymphocytes and monocytes. These populations can be distinguished from one another with monoclonal antibodies that react with unique protein structures on the cell surface designated as the "cluster of differentiation" (CD) antigens. This investigation studied the possibility that the immune abnormalities seen in autism are due to a change in the lymphocytes or monocytes in the subjects with autism . The autistic subjects as compared to age- and sex-matched control subjects exhibited several changes in their cell populations. These included a depression of total lymphocytes, CD2+ (total T cells), CD4+ (helper T cells), and CD4+CD45RA+ (the antigen-inexperienced, suppressor inducer subset of helper T cells). Also analyzed were the siblings of the autistic subjects. A reduced percentage of CD4+ cells was seen in the male siblings as compared to unrelated males. Analyses also compared the cells of mothers and fathers of the autistic subjects with controls of mothers and fathers of normal children. No differences were seen in any of the markers used. Findings in the literature show an increase in memory cells and a decrease in naive cells as a function of age. The data gathered in these experiments uphold this concept and are consistent with the idea that CD45RA and CDw29 are maturational states of helper T cells. The quantitation of different immune markers on lymphoid cells seems to have been useful in the further characterization and investigation of the immune mechanism relevant to the syndrome of autism. Differences in some of the cell types were observed and may account for some of the immune abnormalities seen in autism. These differences may be the result or the cause of the syndrome. Further investigation seems necessary before a direct pathological link can be found between the body's immunity and autism.
6

Diagnosis of haematological malignancies in the era of total laboratory automation: comparison of the Advia 2120 to immunophenotyping and morphology

Pillay, Dashini January 2015 (has links)
A research report submitted to the Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, in partial fulfilment of the requirements for the degree of Master of Medicine in the branch of Haematology. Johannesburg, March 2015 / The incidence of leukaemia in South Africa is 2.5 per 100 000 and has increased due to HIV. Accurate and timeous diagnosis of leukaemia directly impacts success of patient treatment and consequent survival. Usually the Full Blood count (FBC), white blood cell (WBC) differential count and review of the peripheral blood smear alerts the clinician to the possibility of leukaemia. However the number of qualified and skilled technologists in peripheral and central laboratories is on a continual decline making the performance of the critical function of peripheral blood review a challenge. The Advia 2120 haematology analyser performs a WBC and differential count using principles of flow cytometry and the cytograms generated can be used to classify haematological malignancies through the Peroxidase and nuclear density analysis (PANDA) classification system. The presence of myeloperoxidase (MPO) activity in 3% or more leukaemic blasts confirms acute myeloid leukaemia, and enzyme activity can be detected by immunophenotypic analysis or conventional cytochemistry . Research on the comparison of the Advia 2120 and manual morphologic assessment in the classification of leukaemias is limited in the South African setting, where leukaemia often coincides with infection. The aim of this study was to determine if the FBC, differential count and cytogram assessment by the Advia 2120 using the PANDA classification is as reliable as morphologic assessment in the initial classification of haematological malignancies from peripheral blood samples when using flow cytometry as the gold standard.. 150 cases of confirmed leukaemia were collected. The diagnosis obtained from either PANDA analysis and/or morphological assessment was compared to the diagnosis obtained by immunophenotypic analysis. Secondly, the MPO activity obtained by the Advia peroxidase cytogram was compared to the MPO obtained by conventional methods of immunophenotypic analysis and/or cytochemistry. Using the PANDA analysis system, only 48% (72/150) of cases overall were accurately classified. The inaccuracy was 9.3% (14/150) and 42.7% of cases could not be classified. The positive predictive value (PPV) was 88%. The most significant finding was all of the acute Page | iv promyelocytic leukaemia (APL) cases (8/8) had a distinct pattern and were accurately classified on cytogram analysis alone. Accurate sub-classification of other types of acute myeloid leukaemia using PANDA analysis alone was inconsistent. However, the accuracy in classifying leukaemia was improved when the Advia cytogram was used in conjunction with morphological analysis, as 90% (135/150) of cases were accurately classified. The sensitivity and specificity of the peroxidase cytogram in evaluating myeloperoxidase (MPO) activity was 85% and 88.6% respectively. The agreement between cytogram peroxidase activity and the reference methods was 89.1% and the Cohen’s kappa was 76.9%. To the best of our knowledge, there is no data comparing peroxidase activity on the cytogram to other methods. In conclusion, it was shown that the routine use of the Advia cytograms in conjunction with the morphology findings provides invaluable information in the initial screening of leukaemia. In cases with indistinct morphology, the cytograms have the potential to aid in a provisional classification. The peroxidase activity from the cytogram could be used as a surrogate marker for myeloperoxidase activity in leukaemia. Moreover, a tentative diagnosis of an APL is possible by simple analysis of the cytogram resulting in earlier diagnosis which could be life-saving.
7

Immunophenotypic classification of canine malignant lymphoma in formalin-fixed, paraffin wax-embedded specimens using CD3 and CD79a cell markers

Pearson, Joyce 16 November 2006 (has links)
Inconsistent use of nomenclature in different canine malignant lymphoma (CML)classification systems, which lead to incorrect diagnosis and prognosis, necessitated a retrospective study of 103 cases of CML. Histological classification was done according to the Working Formulation, on H&E sections, after standard processing. Immunophenotyping, using CD3 (T cell) and CD79a (8cell) markers, was carried out on the same sections. Intermediate grade lymphomas were the largest category (49.5%), with 16.5%high grade lymphomas. More than half (53.3%) of the lymphomas were of 8 cell immuno phenotype, and 36.5% were T cell lymphomas. Only 9.7% of the total number of lymphomas exhibited double negative staining. Only two categories, the immunoblastic and medium-sized macro nucleolated (MMC) category (Fournel-Fleury et al.). exhibited constant (8 cell) immunophenotype. All the other categories exhibited mixed immunophenotype. The Working Formulation, with omission of the follicular tyoes (due to the rarity thereof in CML) and addition of the MMC category and immunophenotyping, is best for classifying CML. / Dissertation (MMed Vet (Pathology))--University of Pretoria, 1999. / Anatomy and Physiology / Unrestricted
8

Avaliação leucométrica e citofluorométrica do sangue periférico de cães com linfoma, após uso de rhG-CSF, submetidos à alta dose de ciclofosfamida seguida ou não de transplante autólogo de medula óssea

Godoy, Aline Vieira [UNESP] 14 May 2010 (has links) (PDF)
Made available in DSpace on 2014-06-11T19:31:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-05-14Bitstream added on 2014-06-13T19:01:28Z : No. of bitstreams: 1 godoy_av_dr_jabo.pdf: 494650 bytes, checksum: d27d9898dce60f32a0eece2e8cdaf032 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O presente estudo teve como objetivos avaliar seqüencial e temporalmente o quadro leucocitário de dez cães portadores de linfoma em remissão, submetidos à alta dose de ciclofosfamida, durante o uso do estimulador de colônia de granulócitos (GCSF, filgrastine) seguido ou não do transplante autólogo de medula óssea (TMO), bem como quantificar células CD34+ no sangue periférico dos referidos cães. Para tal avaliação foram utilizados três grupos experimentais, sendo o grupo 1 (G1) formado por cinco animais em remissão de linfoma que passaram por alta dose de ciclofosfamida e TMO seguido do uso de G-CSF; o grupo 2 (G2) formado por cinco animais em remissão de linfoma que sofreram alta dose de ciclofosfamida seguida do uso de G-CSF e o grupo 3 (G3) formado por dez animais saudáveis que receberam apenas o G-CSF. O transplante consistiu na colheita de medula óssea, preparo e congelamento da bolsa que continha células medulares em suspensão, condicionamento do paciente com 500mg/m2 de ciclofosfamida, infusão das células hematopoéticas e aplicação do G-CSF. Para avaliar a recuperação hematopoética pós-transplante foi realizado leucograma e análise citométrica do sangue nos dias 7, 8, 9, 10 e 11 após condicionamento. O nadir médio dos neutrófilos segmentados (NS) no grupo com transplante de medula óssea (G1) foi 425 NS/mL, e no grupo sem TMO (G2) foi 637,4 NS/μL e ocorreu sete dias após alta-dose de quimioterápico em ambos os grupos. A duração média da neutropenia foi de três dias. Nenhum animal apresentou febre ou sepse após a alta dose de ciclofosfamida. A dose de 5μg/kg/dia durante quatro dias de filgrastine foi insuficiente na mobilização adequada de células CD34+ nos três grupos estudados, sendo necessários novos estudos para este propósito. Desta maneira, pode-se comprovar o fato de que o uso do G-CSF leva a reduções significativas na incidência... / The aims of this research were to provide an analysis of several counts of leucocytes values in dogs with lymphoma in remission phase, undergone to high-dose chemotherapy with cyclophosphamide, during treatment with granulocyte colonystimulating factor (G-CSF, filgrastine), followed or not by autologous bone marrow transplantation (BMT), as well to quantify CD34+ cells of peripheral blood from that dogs. For this purpose, three experimental groups were considered. Five dogs in clinical remission undergone to high-dose chemotherapy with cyclophosphamide and BMT, followed by G-CSF use were included in group 1 (G1), while group 2 (G2) was consisted by five dogs undergone to high-dose chemotherapy with cyclophosphamide, followed by G-CSF. Group 3 (G3) was composed of ten healthy dogs undergone to GCSF only. Transplantation consisted of bone marrow harvest, managing and freezing bags containing lifted marrow cells, cyclophosphamide conditioning (500mg/m2), hematopoietic cells infusion and treatment with G-CSF. After transplantation, hematopoietic recovery was evaluated by means of leukograms and flow cytofluorimetrical analysis on days 7, 8, 9, 10 and 11 after conditioning. Mean nadir neutrophil (NS) counts in group undergone transplantation (G1) was 425 NS/mL versus 637,4 NS/μL in group without BMT (G2). Nadir was observed on the seventh day after high-dose chemotherapy in both groups and neutropenia mean time was three days. Fever or sepsis was not observed in any dog after high-dose cyclophosphamide. Four days of filgrastine treatment in dose of 5μg/Kg, daily, was not enough to mobilize CD34+ cells appropriately in three groups analyzed, requiring new studies for this purpose. Considering these results, it is possible to conclude that G-CSF significantly reduces the incidence, severity and duration of neutropenia
9

Oberflächenentigen- und Sehnenmarkerexpression equiner multipotenter mesenchymaler Stromazellen / Surface antigen and tendon marker expression in euqine multipotent mesenchymal stromal cells

Päbst, Felicitas Miriam Thekla 09 May 2016 (has links) (PDF)
1. Einleitung Multipotente mesenchymale Stromazellen (MSC) stellen eine interessante Therapieoption in der regenerativen Medizin verschiedener Erkrankungen dar. Aufgrund ihrer Herkunft aus mesodermalem Gewebe ist ihr Einsatz in der Therapie von Sehnenerkrankungen als günstig anzusehen, wo sie bei Pferden bereits erfolgreich verwendet werden. Da dieser Erkrankungskomplex mit degenerativen Veränderungen der Achillessehne des Menschen vergleichbar ist, wäre eine Translation der gewonnenen Ergebnisse in die Humanmedizin wünschenswert. Die zugrunde liegenden Wirkmechanismen bei der Sehnenregeneration sind allerdings bis zum heutigen Tage noch nicht vollständig geklärt. Unter anderem wird eine tenogene Differenzierung der MSC mit nachfolgender Produktion von extrazellulärer Matrix (EZM) diskutiert. Als Nachweis hierfür wird die Genexpression von Matrixproteinen sowie Transkriptionsfaktoren angesehen. Die Isolation von MSC ist aus verschiedenen Geweben möglich; allerdings haben Untersuchungen deutliche Unterschiede in den in-vitro-Charakteristika zwischen den Zellquellen aufgezeigt. Trotz dieser unterschiedlichen Eigenschaften fasst die International Society for Cellular Therapy (ISCT) seit 2006 humane MSC als plastikadhärente Zellen mit tripotentem Differenzierungspotential sowie einem definierten Antigenprofil zusammen. Um eine Vergleichbarkeit equiner und humaner MSC und somit eine bessere Übertragbarkeit gewonnener Erkenntnisse aus der Pferdemedizin zu erreichen, steht aktuell die Untersuchung der geforderten Antigenexpression noch aus. 2. Ziele der Untersuchung In der vorliegenden Arbeit sollte daher erstmalig eine vollständige Charakterisierung des geforderten Antigenprofils equiner MSC aus fünf verschiedenen Quellen durchgeführt werden, um einen Vergleich mit humanen Zellen zu ermöglichen. Zudem sollte eine vergleichende Darstellung der Sehnenmarkerexpression durchgeführt werden, welche das Wissen um die in-vitro-Eigenschaften von MSC erweitern und in Folge zur Auswahl einer optimal für die Therapie von Sehnenerkrankungen geeigneten Zellquelle beitragen soll. 3. Materialien und Methoden In der ersten Studie wurden equine MSC aus Knochenmark, Fettgewebe, Nabelschnurblut, Nabelschnurgewebe und Sehnengewebe bis zur Passage 3 kultiviert und anschließend mittels Durchflusszytometrie auf das Vorkommen der Antigene CD 29, CD 44, CD 73, CD 90 und CD 105 sowie das Fehlen der Antigene CD 14, CD 34, CD 45, CD 79α und MHC II untersucht. In der zweiten Studie wurde eine Genexpressionsanalyse der Sehnenmarker Kollagen 1A2, Kollagen 3A1, Decorin, Tenascin-C und Skleraxis vergleichend mittels Echtzeitpolymerasekettenreaktion an den isolierten Zellen durchgeführt. In beiden Studien wurde eine Probenzahl von n= 6 für jede Zellquelle untersucht. 4. Ergebnisse Keine der untersuchten Zellquellen erfüllte die MSC-Definition der ISCT bezüglich des Antigenprofils. Insbesondere durch den fehlenden Nachweis CD 73 (< 3,07 %) in allen untersuchten Proben unterscheiden sich equine und humane MSC. Die einzigen stabil exprimierten Antigene sind die zusätzlich untersuchten Proteine CD 29 (37,5 % - 65,42 %) und CD 44 (32,2 % - 97,18 %). Das Vorkommen CD 105 konnte in MSC aus Fett- und Sehnengewebe belegt werden. Zusätzlich war ein Nachweis von CD 90 in MSC aus Fettgewebe möglich, welche somit die größte Ähnlichkeit mit der humanen Zellpopulation aufweisen. Die Studie zur Genexpressionsanalyse weist auf eine Basisexpression von Kollagen 1A2, 3A1 und Decorin in MSC aus verschiedenen Quellen hin, welche über der von nativem Sehnengewebe liegt. Auch hier weisen wiederum MSC aus Fettgewebe die höchste Expression auf. 5. Schlussfolgerungen Die vorliegende Arbeit leistet einen Beitrag zu einer vertiefenden in-vitroCharakterisierung equiner MSC. Das Antigenprofil equiner MSC ist nicht vollständig mit dem humaner identisch. Eine abschließende Beurteilung sollte durch Untersuchungen mit spezies-spezifischen Antikörpern erfolgen. Die Ergebnisse der Genexpressionsanalyse unterstützen die Theorie, dass MSC die Sehnenheilung durch Produktion von extrazellulärer Matrix beeinflussen. Der Einsatz von MSC aus Fettgewebe in der Therapie von Sehnenerkrankungen sollte forciert werden, da ihre hohe Sehnenmarkerexpression einen Hinweis auf eine Verbesserung der Sehnenregeneration darstellt.
10

Metabolismo e resposta imune celular no sangue de vacas Holandesas no período de transição / Metabolism and cellular immune response in blood of Holstein cows in the transition period

Baldacim, Vinícius Alvim Passos 28 August 2014 (has links)
O objetivo geral desta pesquisa foi avaliar o metabolismo e a resposta imune celular no sangue de vacas leiteiras no período de transição. Foram utilizadas 13 vacas Holandesas, de 2ª a 4ª parição, avaliadas nas semanas M-2, M-1 (pré-parto), M0 (dia da parição), M1, M2 e M3 (pós-parto). Foram realizadas análises das seguintes variáveis: produção leiteira, escore de condição corporal (ECC), mensuração sérica do beta-hidroxibutirato (BHB), ácidos graxos não esterificados (NEFA), IGF-1 (Fator de Crescimento Semelhante à Insulina Tipo 1), glicose (GLIC), colesterol (COL), triglicerídeos, proteína total (PT), albumina (ALB), globulina (GLOB), AST (Aspartato transaminase), GGT (Gamaglutamiltransferase) e cálcio total e cálcio ionizável. Além disso, a resposta imune celular das vacas forma avaliadas pelas interpretações do leucograma e imunofenotipagem dos linfócitos. Foram encontradas medianas do ECC equivalentes a 4,0; 3,8; 3,5; 3,0; 3,5 e 3,5 dos momentos M-2 ao M3. Em relação aos indicadores energéticos, observou-se aumento da concentração de NEFA e BHB no parto e pós-parto, ao contrário, os teores de triglicerídeos, colesterol e IGF- 1 diminuíram. Além disso, foi possível observar concentração máxima de glicose no momento da parição. Em relação ao metabolismo proteico e hepático, observam-se menores concentrações de proteína total e globulina no momento do parto; os teores de albumina diminuíram no pós-parto. A atividade sérica da AST aumentou a partir da parição, porém não foi possível detectar variações para a GGT. As concentrações de cálcio sério total e ionizado foram menores a partir da parição. A análise do leucograma das vacas no período de transição revelou leucocitose por linfocitose no momento do parto, apesar de não ter sido observada variações no número absoluto e relativo dos linfócitos durante o período de transição, foi possível observar que os linfócitos sanguíneos apresentaram-se elevados durante todo o período de estudo. O aumento no número de linfócitos decorreu da elevação dos linfócitos B (CD21+). A partir desse resultado realizou-se exame sorológico para o vírus da Leucose Enzootica Bovina, detectando 12/13 (92,30%) vacas soropositivas. O linfócito T (CD3+) e suas subpopulções auxiliar (CD3+CD4+) e citotóxica (CD3+CD8+) apresentaram ligeiras oscilações durante o período de estudo. Com base nos resultados obtidos pode-se concluir que: a) As vacas Holandesas apresentaram variações nos parâmetros do perfil energético indicadoras de balanço energético negativo e mobilização lipídica, caracterizados especialmente pela diminuição do ECC e elevações nos teores séricos de NEFA e BHB; b) os teores de PT e GLOB apresentaram variações em decorrência da colostrogênese e infecções uterinas pós-parto. A albumina apresentou diminuição no pós-parto, decorrente do aumento da demanda nutricional para a produção de leite e uso de aminoácidos como precursor energético no processo de gliconeogênese; c) os valores de IGF-I apresentaram acentuada redução no momento do parto, sinalizando para mobilização lipídica e desacoplamento do eixo somatotrópico a partir da parição; d) as vacas apresentaram hipocalcemia, especialmente na parição e primeira semana pós-parto; e) a infecção pelo Vírus da Leucose Enzoótica Bovina influenciou na resposta imune observada no período de transição, caracterizada por linfocitose e aumento da população de linfócitos B (CD21+). / The general aim of this research was to evaluate the metabolism and cellular immune response in blood of dairy cows in transition period. Was evaluated 13 Holstein cows, from 2nd to 4th parturitions in the weeks M-2, M-1 (pre-partum), M0 (parturition day), M1, M2 and M3 (post-partum). Paremeter analysis were performed: milk production, body condition score (BCS), beta-hydroxybutyrate (BHB), nonesterified fatty acids (NEFA), IGF-1 (growth factor Insulin-like similar to insulin type 1), glucose (GLUC), cholesterol (CHOL), triglycerides, total protein (TP), albumin, globulin (GLOB), AST (aspartate transaminase), GGT (gamma-glutamyl transferase), total calcium and ionized calcium. Furthermore, cows cellular immune response was evaluated by WBC and lymphocyte immunophenotyping. Was find for BCS median 4.0; 3.8; 3.5; 3.0; 3.5 and 3.5 between the moments M-2 to M3. Regarding to energy indicators, an increase of NEFA and BHB in parturition and postpartum was observed, in contrast, the values of triglycerides, cholesterol and IGF-1 decreased. Moreover, it was possible to obtain maximum glucose concentration at parturition time. Regarding to protein and hepatic metabolic was find lower values of total protein and globulin was observed in pre-partum and parturition in relation to postpartum; values of albumin decreased in post-partum. The Aspartate aminotransferase activity increased from calving, but it was not possible to detect GGT variations. The concentration of total and ionized calcium serum were lower from calving. The analysis of cows leukogram in the transition period revealed leukocytosis by lymphocytosis at delivery, although no variation in the absolute and relative number of lymphocytes was observed during the transition period. The increase of lymphocytes number occurred due to the increase of B lymphocytes (CD21 +). From this result, serological test for the Enzootic Bovine Leukosis Virus was developed, detecting 12/13 (92,30%) of seropositive cows. T lymphocytes (CD3 +) and subpopulations T helper (CD3+ CD4+) and T cytotoxic (CD3+ CD8+) showed slight fluctuations during period. Based on the results, we can conclude the following: a) Holstein cows showed variations in the parameters of the energy profile indicator of negative energy balance and lipid mobilization, characterized especially by the decreasing of BCS and elevations in serum NEFA and BHB; b) the levels of TP and GLOB showed variations due to colostrogenesis and postpartum uterine infections. The albumin decreased in postpartum, due to increased nutrient requeriment for milk production and use of amino acids as energy precursor in the gluconeogenesis; c) the amounts of IGF-I showed significant reduction at delivery, signaling lipid mobilization and uncoupling of the somatotropic axis after calving; d) cows showed hypocalcemia, especially in calving and first week post-partum; e) Infection with Enzootic Bovine Leukosis Virus influenced the immune response observed in the transition period, characterized by lymphocytosis and the increase of B lymphocytes (CD21 +) population.

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