Diss. (sammanfattning) Stockholm : Karol. inst., 1999. / Härtill 4 uppsatser.

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 Anatomical Pathology Johannesburg, 2016 / Introduction: Plasmablastic lymphoma (PBL) is an uncommon variant of aggressive B-cell non-Hodgkin lymphoma that occurs in immune-compromised individuals, most commonly secondary to HIV infection. This tumour classically occurs in the oral cavity but has also been described in a variety of extra-oral locations. This is a clinicopathological study of 45 cases of extra-oral PBL (EPBL). Aim: To define the clinical parameters, histology and immunophenotypic features of extra-oral plasmablastic lymphoma (EPBL) and assess the extent to which Epstein–Barr virus (EBV) is associated with this tumour. Materials and Methods: This retrospective study on archival cases of EPBL included the patients‟ age, gender, race, HIV status where available and the site of tumour presentation. Of 49 archival cases retrieved, 4 were discounted owing to reclassification as diffuse large B-cell lymphoma (1 case), multiple myeloma or extramedullary plasma cell tumour (3 cases). The remaining 45 cases were reviewed histologically and classified according to whether they displayed a pure plasmablastic (PBm) morphology or a plasmablastic morphology with plasmacytic differentiation (PBm+PCd), and assessed immunohistochemically with CD45 (LCA), CD20, CD79a, PAX5, CD138, MUM1, BLIMP1, VS38c, Ki-67, BCL6, CD10, HHV8 and CyclinD1 using standard automated procedures. The presence of EBV was assessed by chromogenic in-situ hybridisation. Ethical clearance was obtained (M10750 and M120993). Results: 27of the 45 cases had a pure plasmablastic morphology. The remaining 18 cases showed plasmacytic differentiation. There was no site predilection according to histological pattern. 60% of the tumours were reported in males and 40% in females and all were black African patients. The anus was the favoured extra-oral site of presentation (13 of 45 cases, 28%), followed by soft tissue (11 of 45 cases, 24%). There was no significant difference in the age of presentation between males (38.5 years) and females (35.4 years). Of the 18 patients of known HIV status, 17 were HIV positive (94%). The immunohistochemical profile of EPBL recapitulated that found for both oral and extra-oral PBL in the literature, except for CD45 (leucocyte common antigen) which signalled positively in a higher percentage of cases. 36 of 42 cases (85.7%) were positive for CD45. The positive membrane signal for CD45 was of variable intensity, between 5 and 100% of tumour cells. EBV was positive by in situ hybridisation in 37 of 40 cases tested (92.5%). Conclusion: EPBL is identical to its oral counterpart in gender and age distribution, HIV status, morphological appearances, immunophenotypic profile and association with EBV. The high association with EBV as assessed by in-situ hybridisation studies mirrors that of oral-based PBL reported in the literature. EPBL should be regarded as the same tumour as that arising within the oral cavity. A peculiarity observed within this case cohort is the high level of expression of CD45 (leucocyte common antigen). This has been reported to be of low or near absent expression in most cases of PBL, as defined by the 2008 WHO Classification of Tumours of the Haematopoietic and Lymphoid Tissues. / MT2016

Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 5 uppsatser.

Thesis (doctoral)--University of Uppsala, 1984. / Bibliography: p. 59-66.

AIMS OF THE STUDY: 1. To perform a retrospective, epidemiological analysis of cases of reactive lymphadenopathy and atypical reactive lymphoid hyperplasia (ARLH) received in the Department of Anatomical Pathology, UCT and GSH, over a 5 year period, in order to determine the number of cases of ARLH, and the frequency of the various subtypes of reactive lymphoid. hyperplasia, so as to provide base-line information for further studies. 2. To set up IN SITU HYBRIDIZATION (ISH) for detection of Epstein-Barr virus (EBV)-encoded RNA's (EBERs) in latently infected cells in selected cases, to determine if virus is present in ARLH. 3. To perform immunohistochemical analysis for the detection of EBY-derived latent membrane protein (LMP) in those cases subjected to ISH.

by Hon-wing Tsui. / Thesis (M.Ph.)--Chinese University of Hong Kong, 1988. / Bibliography: leaves 151-203.

by Shik Yuen Lo. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 102-124). / Abstract / List of Illustrations / List of Tables / Acknowledgements / Chapter 1. --- Introduction --- p.1 / Chapter 2. --- Literature Review / Chapter 2.1 --- Anatomy of the Human Nasopharynx --- p.4 / Chapter 2.2 --- Histology of the Human Nasopharynx --- p.6 / Chapter 2.3 --- Intra-epithelial Lesions of the Nasopharyngeal Epithelium --- p.10 / Chapter A. --- Hyperplasia / Chapter B. --- Metaplasia / Chapter C. --- Koilocytes / Chapter D. --- Nasopharyngeal intra-epithelial neoplasia / Chapter 2.4 --- Nasopharyngeal Carcinoma --- p.19 / Chapter A. --- Histopathological classification of NPG / Chapter B. --- Epidemiology / Chapter C. --- Etiological factors / Chapter 2.5 --- Epstein-Barr Virus and Nasopharyngeal Carcinoma --- p.27 / Chapter A. --- Serological / Chapter B. --- EBV genome in NPC / Chapter C. --- EBV encoded latent gene products / Chapter 2.6 --- Cancer Genes in Nasopharyngeal Carcinoma --- p.28 / Chapter A. --- "Tumours suppressor Gene, p53" / Chapter B. --- "Oncogenes, c-myc, ras and bcl-2" / Chapter 2.7 --- Immunohistochemical methods --- p.33 / Chapter A. --- Avidin-Biotin Complex method (ABC) / Chapter B. --- Alkaline phosphotase Anti-alkaline phosphotase method (APAAP) / Chapter C. --- Unmasking of antigens / Chapter 2.8 --- Techniques in ISH --- p.40 / Chapter 3. --- Material and Methods --- p.42 / Chapter 3.1 --- Tissue Samples --- p.42 / Chapter A. --- "Samples for ras, c-myc and p53 studies" / Chapter B. --- Samples for LMP-1 study / Chapter C. --- Samples for bcl-2 study / Chapter D. --- Samples for EBER-RNAs study / Chapter 3.2 --- Monoclonal Antibodies --- p.47 / Chapter 3.3 --- Tissue Processing --- p.49 / Chapter A. --- Tissue processing for formalin fixed tissue / Chapter B. --- Tissue processing for frozen section / Chapter 3.4 --- IHC Techniques --- p.50 / Chapter A. --- Pretreatment of Laboratory Wares / Chapter B. --- Determination of optimum dilution and incubation time for p53antibody / Chapter C. --- Determination of optimum dilution and incubation time for bcl-2 and LMP-1 antibodies / Chapter D. --- Determination of optimum dilution and incubation time for c-myc and ras / Chapter E. --- "Detection of p53, c-myc and ras by ABC method" / Chapter F. --- Detection of bcl-2 and LMP-1 by APAAP method / Chapter 3.6 --- ISH --- p.57 / Chapter A. --- Pretreatment of laboratory wares / Chapter B. --- FITC conjugated EBER oligonucleotide probe / Chapter C. --- Determination of PK dilution for paraffin section / Chapter D. --- Determination of PK dilution for frozen section / Chapter E. --- Determination of the choice of fixative for frozen section / Chapter F. --- Detection of EBER-RNAs by ISH method / Chapter 3.7 --- Statistical analysis --- p.62 / Chapter A. --- p53 / Chapter B. --- c-myc and ras / Chapter 4. --- Results --- p.63 / Chapter A. --- ras / Chapter B. --- c-myc / Chapter C. --- p53 / Chapter D. --- LMP-1 / Chapter E. --- Bcl-2 / Chapter F. --- EBER-RNAs / Chapter 5. --- Discussion --- p.86 / Chapter 6. --- Conclusion and Summary --- p.97 / Appendix --- p.99 / Reference --- p.102

by Suet-ching Leung. / Thesis (M.Ph.)--Chinese University of Hong Kong, 1989. / Bibliography: leaves 170-198.

by Ka-chun Yiu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1989. / Bibliography: leaves 162-192.

Ng Dean Yew, Dennis. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 85-98). / Abstract --- p.i / Acknowledgments --- p.iii / Table of contents --- p.iv / List of figures --- p.vii / List of tables --- p.ix / List of abbreviation --- p.x / Chapter Chapter 1 --- Introduction / Chapter 1.1. --- History --- p.1 / Chapter 1.2. --- Classification and structure of Epstein-Barr Virus --- p.2 / Chapter 1.3. --- Genomic organization of EBV --- p.3 / Chapter 1.4. --- Replication cycle of EBV --- p.5 / Chapter 1.5. --- EBV latent and lytic cycle proteins --- p.6 / Chapter 1.6. --- Clinical diseases associated with EBV Infection --- p.11 / Chapter 1.7. --- Association of EBV and NPC --- p.13 / Chapter 1.8. --- EBV serological markers in the diagnosis of NPC --- p.13 / Chapter 1.9. --- Sources of EBV-specific DNase --- p.15 / Chapter 1.10. --- Characteristics of Epstein-Barr virus alkaline DNase --- p.15 / Chapter 1.11. --- Aim of the project --- p.18 / Chapter Chapter 2 --- Materials & Methods / Chapter 2.1. --- Molecular cloning --- p.19 / Chapter 2.1.1. --- Cell culture --- p.19 / Chapter 2.1.2. --- mRNA purification --- p.19 / Chapter 2.1.3. --- First strand cDNA synthesis --- p.21 / Chapter 2.1.4. --- Polymerase chain reaction (PCR) of cDNA --- p.21 / Chapter 2.1.5. --- Purification of PCR product after gel electrophoresis --- p.22 / Chapter 2.1.6. --- Ligation of PCR amplified DNase gene into pUC18 Sma/BAP vector --- p.23 / Chapter 2.1.7. --- Transformation by electroporation --- p.24 / Chapter 2.1.7.1. --- Cell preparation --- p.24 / Chapter 2.1.7.2. --- Electroporation procedure --- p.25 / Chapter 2.2. --- Extraction ofplasmid DNA --- p.28 / Chapter 2.2.1. --- Boiling preparation --- p.28 / Chapter 2.2.2. --- Plasmid digestion --- p.29 / Chapter 2.3. --- Large-scale purification ofplasmid --- p.29 / Chapter 2.4. --- Small-scale purification ofplasmid --- p.32 / Chapter 2.5. --- DNA sequencing --- p.33 / Chapter 2.5.1. --- Annealing of primer to template DNA --- p.33 / Chapter 2.5.2. --- Labelling reaction --- p.34 / Chapter 2.5.3. --- Sequencing termination reaction --- p.35 / Chapter 2.5.4. --- Prepartion of sequencing gel --- p.36 / Chapter 2.5.5. --- Autoradiography of sequencing gel --- p.38 / Chapter 2.6. --- Epitope mapping --- p.39 / Chapter 2.6.1. --- Processing of EBV- specific DNase peptides --- p.39 / Chapter Chapter 3 --- Results / Chapter 3.1. --- Molecular cloning --- p.41 / Chapter 3.1.1. --- Cell culture --- p.41 / Chapter 3.1.2. --- mRNA purification --- p.42 / Chapter 3.1.3. --- PCR amplification --- p.42 / Chapter 3. 1.4 --- DNA purification of PCR product --- p.42 / Chapter 3.1.5. --- Molecular cloning of PCR amplified DNase gene into pUC18 SmaI/BAP vector --- p.44 / Chapter 3.1.6. --- Transformation by electroporation --- p.46 / Chapter 3.1.7. --- Extraction of plasmid DNA --- p.48 / Chapter 3.1.7.1. --- Boiling preparation --- p.48 / Chapter 3.1.8. --- Plasmid digestion --- p.51 / Chapter 3.2. --- DNA sequencing --- p.51 / Chapter 3.2.1. --- Comparison of B95-8 EBV-speicific DNase gene with gene sequence of EBV in GeneBank --- p.50 / Chapter 3.2.2. --- Comparison of 5' end of Raji & B95-8 EBV derived EBV-specific DNase gene --- p.57 / Chapter 3.2.3. --- Comparison of the 3'end of the Raji and B95-8 denved EBV-specific DNase gene --- p.63 / Chapter 3.2.4. --- Amino acid sequence homology between B95-8 & Raji EBV-specific DNase --- p.64 / Chapter 3.2.5. --- Amino acid sequence comparison between the 3' end of the B95-8 EBV DNase protein with that of the Raji EBV DNase protein --- p.62 / Chapter 3.3. --- Epitope mapping --- p.67 / Chapter 3.3.1. --- Amino acid key --- p.67 / Chapter 3.3.2. --- Amino acid sequence of peptides --- p.73 / Chapter 3.3.2. --- O.D. readings at 492nm of five histologically proven NPC sera --- p.74 / Chapter Chapter 4 --- Discussions / Chapter 4.1. --- Overall strategy --- p.75 / Chapter 4 2 --- Significance of EBV-specific DNase as marker for NPC --- p.76 / Chapter 4.3. --- Characterization of EBV-specific DNase --- p.76 / Chapter 4.4. --- Molecular cloning of PCR amplified gene into PUC18 SmaI/BAP vector --- p.77 / Chapter 4.4.1. --- Cell culture --- p.77 / Chapter 4.4.2. --- PCR amplification --- p.73 / Chapter 4.4.3. --- "Blunting,kinasing and ligation of EBV-specific DNase cDNA into pUC18 vector" --- p.78 / Chapter 4.4 .4 --- .Transformation by electroporation --- p.80 / Chapter 4.4.5. --- Restriction enzyme digestion of pUC18/EBV-DNase plasmid … --- p.81 / Chapter 4.5. --- DNA sequencing --- p.81 / Chapter 4.6. --- Epitope mapping --- p.83 / Reference --- p.85