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A thesis submitted to the Faculty of Medicine. University of the Witwatersrand. Johannesburg, for the Degree of Doctor of Philosophy / Abnormal cell membrane function and altered cellular cation homeostasis have been implicated in the pathogenesis of essential hypertension. The role of these factors in the aetiology of malignant hypertension is unknown. (Abbreviation abstract) / Andrew Chakane 2019

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Class of 2012 Abstract / Specific Aims: To evaluate disease- and patient-related characteristics, mortality, and charges associated with non-AIDS defining malignancies (NADM) among inpatient settings in the United States from 2005 to 2009. Methods: This retrospective cohort investigation utilized nationally-representative hospital discharge records from the Agency for Healthcare Research and Quality (AHRQ) Healthcare Cost and Utilization Project (H-CUP) Nationwide Inpatient Sample. Inclusion criteria included adult inpatients ≥18 years with a diagnosis of HIV or AIDS and malignant neoplasms. Multivariate regression analyses were used to assess inpatient mortality and charges. Main Results: Overall, 104,488 were included. Average age associated with each case was 46.9 years (±10.66), with 21.9% cases being female (n=22,868). The mean length of stay was 8.6 days (±10.5) and inpatient mortality occurred in 7.7% of cases (n=8,035). The mean number of procedures performed was 2.3 (±2.5) and the mean number of diagnoses on record was 9.5 (±4.4). Charges for each episode of care averaged $59,483 (±85,748), summing to a national bill of $6.14 billion (2011 dollars) over the five-year course. A higher number of cases were associated with teaching hospitals (74.1%), the south (42%), large metropolitan areas (75.1%), median household income in the 0-25th percentile (41.2%), and Medicaid payers (34.3%). Increased mortality was associated with increased age, increased number of diagnoses and procedures, and the comorbidities of anemia, coagulopathy, lymphoma, and fluid and electrolyte disorders. Conclusions: This investigation of NADMs suggest a considerable clinical and economic burden of illness, summing to a 7.7% inpatient death rate and $1.3 billion in charges per year.

Thesis (M.A.)--Boston University / In view of the success of the Danish investigators Christensen and Kjems in inducing regressions of malignant tumors with streptococcus bacteriophage lysates, it was decided to test lysates of hemolytic Staphylococcus aureus upon malignancies in hamsters and mice. [TRUNCATED]

Malignant mesothelioma (MM) is an aggressive and highly chemo-resistant tumour of the mesothelium. Asbestos is indicated as the environmental factor most commonly associated with mesothelioma. The chemo-resistance is possibly due to impaired apoptotic mechanisms since it is known most chemotherapeutic drugs act via apoptosis. Murine MM cell lines that have been derived from tumours induced by inoculation of crocidolite asbestos into mice provide a suitable model, since both phenotypic and biological properties are closely similar to the human disease. Four murine MM cell lines were used in this study, namely ABl, AB12, AC29 and AC34. The aim of this study was to determine susceptibility of those MM cell lines to induction of apoptosis and the expression of key molecules in those cells. Many apoptosis-related genes are known, expression of some of these genes in four murine MM cell lines were investigated in this study. Gene expression was determined using conventional reverse transcription PCR (RT-PCR) and quantitated using real-time RT-PCR with SYBR-Green I detection on a Rotor-Gene 2000 (Corbett Research, N.S.W., Australia). Gene expression data was normalised against the most stable housekeeping genes as determined by the geNonn software. Susceptibility of the four murine MM cell lines to apoptosis induction was determined using cisplatin or TNF-a or IFN-y at different concentrations and at different times in the case of cisplatin. Apoptosis was assessed by a DNA laddering assay. Conventional RT-PCR results showed that all four murine MM cell lines expressed DR5, Bax, Bcl-xL, FLIP-L, c-Myc and caspase-3. Fas mRNA was detected in all cell lines except AC29. Neither FasL nor Bcl-2 was expressed in the four murine MM cell lines. / Quantitation of gene expression showed that there were significant differences in Fas, DRS, Bax, Bcl-xL, c-Myc, FLIP-L and caspase-3 mRNA levels across the cell lines (P<0.05). Absence of Fas receptor in AC29 may play a role in the immunogenicity of this cell line. DNA laddering results indicated that cisplatin induced apoptosis in a dose- and time- dependent manner in those MM cell lines. Susceptibility as reflected by the minimum apoptosis-inducing dose varied among the cell lines from 1 pdml (AB12) to 10 & n l (AC34). Susceptibility to TNF-a or IFN-y also varied among the cell lines where AB12 was sensitive while AC29 was resistant to those cytokines. The AC29 and AB1 cell lines were used to examine cisplatin or TNF-a induced genes related to apoptosis, expression of Fas, caspase-3, Bax and Bcl-xL mRNA was determined using real-time RT-PCR after 6 and 24 hours induction with cisplatin or TNF-a. The results demonstrated that caspase-3 and Bax were up-regulated whereas Bcl-xL was down-regulated in AC29 after 6 hours treatment with cisplatin. Although only Bcl-xL was down-regulated in ABl after 6 hours treatment with cisplatin, the down regulation was more pronounced than in AC29. TNF-a induction of gene expression showed that Bcl-xl decreased and Fas was up-regulated significantly in AB1 after 6 hours whereas only Bcl-xL was down-regulated in AC29 after 6 hours and continued to decrease until 24 hours. The differences in gene expression changes were noticed not only between cell lines but also between two inducer agents. There were several significant results of this study. Firstly, gene expression in murine MM was seen to parallel those that have previously been described for the human disease. / Therefore murine MM is likely to be a useful in in vivo model for future studies targeting apoptotic molecules. Secondly, a number of genes were not previously examined in MM were characterized in the murine model. Finally, differences in basal and induced gene expression between cell lines and inducing agents were characterized which should be followed in further studies at the protein level (eg caspase-3 activity).

Because of the failure so far to find effective treatment for patients with advanced stages of melanoma, increasing efforts have been made to find prognostic factors identifying patients in the risk zone for development of metastasis. This thesis investigates the prognostic powers of a few selected serological and immunohistochemical biomarkers. In the first and second study, patients operated on for localized malignant melanoma were investigated regarding the prognostic impact of angiogenic serological markers and circulating levels of S100. We concluded that the S100 assays, especially S100BB, are potential biomarkers in patients with malignant melanoma, correlated to both survival and disease free survival. However, no such conclusion could be drawn from the first study, where we found no correlation to survival and investigated angiogenic markers. In the third and fourth study four new potential immunohistochemical biomarkers where investigated in collaboration with the Swedish Human Protein Atlas Program, and those where TRP-1, galectin-1, DLG5 and syntaxin-7. We found that TRP-1 correlated inversely with tumor stage and galectin-1 correlated to Ki-67. DLG5 showed a significant inverse correlation to Ki67 and the expression of STX7 was inversely correlated to tumor stage, suggesting that decreased expression is associated with more aggressive tumors. None of the investigated markers in study III and IV correlated with disease free survival or overall survival. In the fifth and last study, we examined the expression of SOX10, a transcription factor, in different melanocytic lesions. Also, a proliferation assay was carried out in a human melanoma cell line. The results reveal the presence of SOX10 in different melanocytic lesions, with a weak inverse correlation to survival and a significant inverse correlation to T-stage. A significant decrease in proliferation rate for SOX10 silenced cells was found and our data also suggests an increased migratory response in SOX10 silenced cells.

Cancer has become the first among the 10 major death causing factors in Taiwan. Glioblastoma multiforme (GBM) is the most common malignant tumor in adult human brain tumors. Previously, heat shock or all trans retinoic acid (ATRA) treatment has been shown to be effective in inducing cell apoptosis and cell cycle arrest in several cancer cell lines. In this study, human brain tumor cell line GBM8401 was exposed to 43¢J for 30 min followed by incubation with ATRA. The treatment resulted in up to 50% inhibition of cell growth rate and 50% reduction of cell survival rate . Analysis of cell apoptosis related gene expression and protein expression with RT-PCR and Western blot has showen that p21, p27, pro-caspase 3, phospho-JNK and phospho-p38 were overexpressed after treatment of tumor cells with 43¢J for 30 min followed by addition of ATRA for 15 min to 8 hr. The immunocytochemistry assay revealed that overexpression of phospho-p53 in the nuclei after tumor cells were treated with 43¢J for 30 min followed by addition of ATRA for 8 hr. Results from this study show that treating tumor cells with heat shock before incubation with ATRA will enhance cell apoptosis and inhibit cell growth.