Masters of Science / The origin of HIV/AIDS has puzzled scientists ever since the illness first became known in the early 1980s. It is now over twenty years down the line, still the pandemic is the subject of fierce debate and the leading cause of death in the world with sub-Saharan Africa being the worst hit region. With almost everything put in place in 21st century, the rates of infection continues to rise, thus this study tries to find out the undermining factors for full utilization of HIV services in higher institutions of learning,particularly the University of the Western Cape.The services are; free HIV testing and counseling, free medication; provision of free condoms to the students in their halls of residences, peer education programs, and making them easy to access, and prevention of mother to child transmissions.This qualitative study was conducted from June to November 2009, using designed questionnaires for sixty three (63) registered students and five (5) HIV program staff .The main reason for this study was to understand the underlying factors for why students may utilize or may not utilize the available HIV services on campus. The willingness of students to express their views was a positive finding in this study. Majority students who answered the questionnaires were quite aware of these HIV services. They also agreed that services provided are good. The study also found out that females utilized these services more than males and majority of students learnt of the HIV services from the HIV programs pamphlets and website thus indicating that the HIV program at UWC is function. However the research study also found out that the though students are aware of these services few utilize them and majority are females thus leaves a question why males do not utilize.

Dopamine receptors belong to the G protein-coupled receptor (GPCR) superfamily and are classified into two families: D1-like (D1R and D5R) and D2-like (D2R, D3R and D4R), based on their ability to stimulate or inhibit adenylyl cyclase (AC). Classically, GPCRs (including D2R and D3R) are desensitized by the activation of the serine/threonine protein kinase C (PKC) upon phorbol-12-myristate-13-acetate (PMA) treatment. Previous studies demonstrate that while human D5R (hD5R) is also strongly desensitized upon PMA treatment, the human D1R (hD1R) undergo a robust PMA-induced sensitization. The aim of this PhD thesis was to explore how the canonical PKC- or phorbol ester-linked pathway can control the responsiveness of two similar GPCRs like hD1R and hD5R in an opposite fashion. Our data indicate that hD1R sensitization and hD5R desensitization are not mediated by a direct modulation of AC activity by PKC. Using a chimeric approach, we identified the third intracellular loop (IL3) as the key structural determinant controlling in an opposite manner the PMA-mediated regulation of hD1R and hD5R. To delineate the potential PKC phosphorylation sites, a series of mutation of serine (Ser) and threonine (Thr) located into IL3 of hD1R and hD5R were used. No hD1R mutation decreased the PMA-mediated sensitization. This suggests that hD1R phosphorylation is not required for PMA-induced sensitization. In contrast, our results indicate that PMA-mediated hD5R desensitization occurs through a hierarchical phosphorylation of Ser260, Ser261, Ser271 and Ser274. Notably, these hD5R mutants exhibited a PMA-induced sensitization, reminiscent of the PMA-induced hD1R sensitization. Additionally, using short hairpin RNAs (shRNAs), we showed that PKCε is the potentiating PKC while the desensitizing isoform is δ. Overall, our work suggests the presence or absence of specific Ser residues on IL3 of hD1-like receptors dictate if phosphorylation-dependent desensitization (through PKCδ) or phosphorylation-independent potentiation (via PKCε) will occur.

The major topics discussed are the phosphorescence sensitization in the lanthanides via energy transfer and in the organics by heavy atom effects. The f-f transitions in lanthanides are parity forbidden and have weak molar extinction coefficients. Upon complexation with the ligand, ttrpy (4'-p-Tolyl-[2,2':6',2"]-terpyridine) the absorption takes place through the ligand and the excitation is transferred to the lanthanides, which in turn emit. This process is known as "sensitized luminescence." Bright red emission from europium and bright green emission from terbium complexes were observed. There is ongoing work on the making of OLEDs with neutral complexes of lanthanide hexafluoroacetyl acetonate/ttrpy, studied in this dissertation. Attempts to observe analogous energy transfer from the inorganic donor complexes of Au(I) thiocyanates were unsuccessful due to poor overlap of the emissions of these systems with the absorptions of Eu(III) and Tb(III). Photophysics of silver-aromatic complexes deals with the enhancement of phosphorescence in the aromatics. The heavy atom effect of the silver is responsible for this enhancement in phosphorescence. Aromatics such as naphthalene, perylene, anthracene and pyrene were involved in this study. Stern Volmer plots were studied by performing the quenching studies. The quenchers employed were both heavy metals such as silver and thallium and lighter metal like potassium. Dynamic quenching as the predominant phenomenon was noticed.

Background: Nuclear and conventional weapons industry workers are at risk for exposures to beryllium, asbestos, high explosives and barium, all of which are implicated in the pathogenesis of pneumoconiosis. Beryllium has also been shown to cause sensitization (BeS) carrying a risk of progression to Chronic Beryllium Lung Disease (CBD). Data are lacking on the epidemiology of beryllium related health effects in conventional munitions workers and limited studies have been published on the prevalence of BeS in workers with minimal exposure. Data on the prevalence of pneumoconiosis in nuclear weapons workers is also lacking. The main objectives of this study were to determine prevalence and risk factors for beryllium sensitization in former nuclear and conventional munitions workers and rates of and risk factors for pneumoconiosis in former nuclear weapons workers, both cohorts from the Iowa Army Ammunition Plant (IAAAP) in Burlington, IA. Methods: Former nuclear weapons workers were offered chest x-ray (CXR) and blood screening for sensitization with beryllium lymphocyte proliferation test (BeLPT) as part of the Department of Energy (DoE) Former Worker Medical Screening Program. Conventional munitions workers were offered BeLPT and clinical follow-up if sensitized, as part of a Department of Defense (DOD) funded study. Chest x-rays were reviewed by three readers according to the International Labour Organization's Classification system for Radiographs for Pneumoconioses (ILO system). Exposures under study were characterized qualitatively by the industrial hygiene team and based on former worker interviews and historical industrial hygiene records. Results: The prevalence of beryllium sensitization in nuclear and conventional munitions workers was found to be slightly higher than in other workforces and weapons worker populations at low risk for exposure. The prevalence of parenchymal disease was higher in these nuclear weapons workers than in other DoE studies, while the prevalence of coincident parenchymal and pleural and isolated pleural disease was lower than in other nuclear weapons populations. Workers who occasionally dressed the copper-beryllium alloy tools were found to have an increased risk of beryllium sensitization, compared to those in administrative or other jobs with insignificant potential for exposure on site. Exposure to beryllium, asbestos, high explosives or barium was not associated with lung disease in this population. Conclusions: The findings from this study have potential policy implications for DOE and DOD to extend or implement beryllium surveillance and lung disease screening for their workforces and better control use of the copper-beryllium alloy tools in their production processes.

京都大学 / 0048 / 新制・課程博士 / 博士(医科学) / 甲第22122号 / 医科博第107号 / 新制||医科||7(附属図書館) / 京都大学大学院医学研究科医科学専攻 / (主査)教授 生田 宏一, 教授 濵﨑 洋子, 教授 杉田 昌彦 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Focused ultrasound (FUS) is a less-invasive medical technique with the potential to improve the treatment outcome of many diseases by utilizing acoustic transducers to generate and concentrate the multiple intersecting ultrasonic waves on a targeted site in the body. The bio-effects induced by FUS are mostly classified into thermal and mechanical effects (mainly focus on cavitation effect). Cavitation is capable of disrupting tumor vasculature and cell membranes. Numerous studies reported that cavitation-induced sonoporation could provoke multiple anti-proliferative effects on cancer cells, including cell-cycle arrest, cell apoptosis, and clonogenicity suppression. Therefore, the combination of FUS-induced cavitation and other treatment modalities like radiation therapy is of great interest, but research in this field is inadequate. A special high-throughput FUS system was used for cancer cell treatment with a customized 1.467 MHz single focused transducer. Characterization of acoustic behavior of gas-filled cavities was performed via a fiber-optic hydrophone (FOH) system and chemical terephthalic acid method helped to define the acoustic parameters, which could lead to occurrence of cavitation at the bottom of 96-well cell culture plates where cancer cells were located. Cavitation occurs at and above the acoustic intensity of 344 W/ cm2 for the 1.467 MHz transducer. The short- and long-term effects of FUS-induced cavitation and adjuvant effects to radiation therapy, standard hyperthermia and testosterone treatment (only for prostate cancer) were investigated comprehensively at the cellular and molecular levels in human prostate cancer (PC-3 and LNCap), glioblastoma (T98G) and head and neck (FaDu) cells in vitro. The long-term additive effects of short FUS shots (with or without cavitation) to radiation therapy (RT) or hyperthermia (HT) were displayed by significantly reduced clonogenic survival in PC-3, T98G and FaDu cells compared to single treatments. The combination treatment of short FUS with cavitation (FUS-Cav) and RT led to a comparable radio-sensitization effect to HT at 45 °C for 30 min and showed a significant reduction in treatment duration, especially for PC-3 cells. The short-term additive effects of short FUS shots to RT or HT are manifested in reducing the potential of cells to invade and decreased metabolic activity. The induction of sonoporation by FUS-Cav was supposed to be the mechanism of cancer cell sensitization to other therapies at the cellular level. The dramatic decline of 5α-reductase type III (SRD5A3) level induced by combination treatment with FUS-Cav and HT is presumed as the underlying mechanism of additive effects of FUS-Cav to HT at the molecular level. Besides, testosterone solutions with normal physiological levels were discovered to inhibit the long-term metabolic activity of androgen-dependent prostate cancer cells LNCap in vitro, while short FUS shots displayed a long-term additive effect to the testosterone treatment. The presented multilevel study demonstrates that short FUS shots using FUS-induced cavitation carry the potential to sensitize cancer cells to other cancer treatment modalities precisely and less-invasively, providing a promising adjuvant therapy to cancer treatments in the future.:1 Abbreviations 2 Summary 3 Introduction 4 Medical and technical background 4.1 The biological basis of prostate cancer treatment 4.1.1 Androgen receptor: an essential signaling pathway for progression of prostate cancer 4.1.2 5α-reductase: a promising therapeutic target for prostate cancer therapy 4.1.3 Testosterone: duality effects in prostate cancer development 4.2 Advantages and disadvantages of current clinical treatments of prostate cancer 4.3 Basics of focused ultrasound (FUS) 4.3.1 Medical application of FUS-induced thermal effects 4.3.1.1 High-intensity focused ultrasound (HIFU) induced thermal ablation 4.3.1.2 Hyperthermia: an alternative heating strategy to sensitize cancer cells for radiation therapy and chemotherapy 4.3.1.3 FUS-induced hyperthermia triggered drug delivery with thermo-sensitive drug carriers 4.3.2 Medical application of FUS-induced mechanical/cavitation effects 4.3.2.1 Cell sonoporation for drug delivery 4.3.2.2 Sonoporation induced anti-proliferative effects for cancer cells 4.3.2.3 Histotripsy 4.3.2.4 Anti-vascular and anti-metastatic effects 4.3.3 The state of art of cavitation detection in medical application 4.3.3.1 Sonoluminescence and sonochemistry 4.3.3.2 Passive cavitation detection 4.3.3.3 Active cavitation detection 4.3.3.4 High-speed sequential photography of cavitation dynamics 4.3.3.5 Laser scattering technique 4.3.3.6 Synchrotron X-ray imaging technique 4.3.3.7 MRI techniques 5 Aims of the thesis 6 Materials and methods 6.1 Materials 6.1.1 Devices 6.1.2 Chemicals and reagents 6.1.3 Consumables 6.1.4 Human cancer cell lines 6.2 Methods 6.2.1 Composition of the FUS system for in vitro treatment of cells 6.2.2 Physical characterization of the in vitro FUS system 6.2.2.1 Setup of fiber-optic hydrophone system to characterize the FUS apparatus 6.2.2.2 Data analysis in MATLAB 6.2.3 Cavitation measurement with FOH system 6.2.4 Chemical cavitation measurement with terephthalic acid (TA) 6.2.5 Culture of human cancer cell lines 6.2.6 FUS treatment of cancer cells 6.2.7 Water bath hyperthermia treatment 6.2.8 Radiation therapy in vitro 6.2.9 The protocol of FUS\FUS-Cav combined with RT or HT 6.2.10 Evaluation of cell ability to reproduce with clonogenic assay 6.2.11 Measurement of cellular metabolic activity with WST-1 assay 6.2.12 Evaluation of cell invasion ability with Transwell® assay 6.2.13 Detection of sonoporation by cell staining with propidium iodide (PI) 6.2.14 Detection of SRD5A as a therapeutic target for prostate cancer with immunofluorescence microscopy 6.2.15 Quantification for the reduction of SRD5A proteins with flow cytometry 6.2.16 Testosterone treatment 6.2.17 FUS/FUS-Cav combined with testosterone treatment 7 Results 7.1 Physical characterization of the in vitro FUS system 7.2 Cavitation occurs at a certain level of acoustic intensity 7.2.1 Characteristics of ultrasonic spectrograms 7.2.2 Cavitation dose depends on the acoustic intensity 7.3 FUS/FUS-Cav supports RT to reduce the long-term clonogenic survival of cancer cells 7.4 FUS/FUS-Cav increases the effects of HT by reducing the long-term clonogenic survival of cancer cells 7.5 FUS/FUS-Cav enhances the suppressive effects of RT in short-term cell potential to invade and metabolic activity of prostate cancer cells 7.6 FUS/FUS-Cav supports HT to diminish the short-term cell potential to invade and metabolic activity of prostate cancer cells 7.7 FUS-Cav treatment immediately induces sonoporation effects in PC-3 and FaDu cells 7.8 FUS/FUS-Cav enhances the effects of HT by inhibiting the SRD5A protein level in prostate cancer cell lines 7.9 FUS-Cav enhances the effects of the testosterone treatment by reducing the long-term cell metabolic activity of androgen-dependent prostate cancer cell line 8 Discussion 8.1 Cavitation measurement in a defined 96-well plate by PCD technique and sonochemistry method 8.2 Short-term and long-term additive effects of FUS-Cav to RT or HT 8.3 Inhibitory effects of FUS-cav in the potential of prostate cancer cells to invade 8.4 The reduction of the long-term metabolic activity of androgen-dependent prostate cancer cells by the combination treatment of FUS-Cav and testosterone 9 Conclusion 10 References 11 Appendix 11.1 Erklärung über die eigenständige Abfassung der Arbeit 11.2 List of figures 11.3 List of tables 11.4 Curriculum vitae 11.5 Acknowledgments

To determine whether behavioral sensitization produced by prolonged D-amphetamine administration affects susceptibility of nigrostriatal dopaminergic neurons to the neurotoxic actions of 6-hydroxydopamine (6-OHDA), rats were treated daily from the 23 rd day after birth for 11 consecutive days with D-amphetamine (1.0 mg/kg s.c.) or saline. On the last day of treatment, one group primed with D-amphetamine and one control group of rats were tested to confirm behavioral sensitization development. The remaining animals were additionally treated on the 34 th day (one day after the last D-amphetamine injection) with 6-OHDA HBr (300 μg in 10 μl i.c.v., salt form, half in each lateral ventricle) or its vehicle. Four weeks later the levels of dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), 3-metoxytyramine (3-MT), as well as 5-hydroxytrypatmine (5-HT) and its metabolite 5-hydroxyindoleacteic acid (5-HIAA) were assayed in the striatum, by HPLC/ED. In rats with behavioral sensitization, 6-OHDA reduced endogenous dopamine and its metabolites content to a comparable degree in comparison to controls. This finding indicates that presumed up-regulation of the dopamine transporter in the behaviorially sensitized rats did not increase the neurotoxicity of a high dose of 6-OHDA.

In agreement with previous work, adult rats given selective lesions to dopamine (DA)-containing neurons as neonates exhibited a greater behavioral sensitization to repeated phencyclidine (PCP) treatment in comparison to sham-lesioned controls. Acute administration of olanzapine (1-5 mg/kg ip) or clozapine (15 mg/kg ip) decreased sensitized PCP-induced activity in both lesioned and control animals. Acute haloperidol (0.5 mg/kg ip) had no impact on PCP responsiveness in lesioned animals, but significantly antagonized PCP effects in sham-lesioned controls. Ketanserin, a selective 5-HT 2A/5-HT2C-receptor antagonist, significantly reduced PCP activation in both lesioned and control rats, suggesting that the efficacy of atypical antipsychotics against PCP-induced sensitized responses may be mediated by one of the 5-HT2-receptor subtypes. A 6-week chronic regimen of orally administered olanzapine, clozapine, or haloperidol failed to block the sensitization induced by repeated PCP exposure. However, a 10-month oral olanzapine treatment significantly blunted the behavioral sensitization to repeated PCP exposure in lesioned animals, even after withdrawal from chronic olanzapine for more than 3 weeks. A 10-month oral haloperidol treatment had no effect on the sensitization induced by repeated PCP dosing. The persistent effect of chronic olanzapine administration on PCP sensitization may be relevant to the chronic therapeutic efficacy of atypical antipsychotics treating schizophrenia - a clinical syndrome linked to enhanced sensitivity to N-methyl-D-aspartate (NMDA)-receptor antagonists.

Hypersensitivity after tissue injury is an expression of neuronal plasticity in the central nervous system. This has been explored most extensively using in vitro preparations and animal models of inflammatory pain and chemical irritation. For pain after surgery, a similar process has been proposed. In the present study, we examined dorsal horn neuron (DHN) sensitization using the plantar incision model for post-operative pain. In behavioral experiments, the effect of a local anesthetic injection (or saline vehicle) 15min before plantar incision on pain behaviors several days after incision was studied. Bupivacaine injection before incision prevented pain behaviors until 4h afterwards; injection after incision produced the same effect. One day after incision, pain behaviors were not different between rats injected with saline or bupivacaine. In neurophysiologic experiments, however, bupivacaine injection blocked activation of DHNs during incision. One hour after incision, expansion of receptive fields (RFs) to pinch and increased background activity occurred in 14 of 16 neurons in the saline group but only in two of 22 neurons in the bupivacaine group. The difference was not due to a systemic effect of bupivacaine. Ten sensitized neurons were studied using the injection of bupivacaine 90min after incision. Increased background activity (n=7) and expanded RFs (n=7) were reversed by bupivacaine. Sensitization was re-established in seven of eight neurons 2h after injection as the local anesthetic dissipated. These results indicate that activation of DHNs during plantar incision and sensitization 1h later are not necessary for subsequent pain behaviors. Because sensitization was reversed 90min after plantar incision and then re-established as the local anesthetic effect diminished, enhanced responsiveness of DHN requires ongoing afferent input during the first day after incision.

This study investigated the effects of repeated administrations of nicotine (0.7 mg/kg) on dendritic morphology in the nucleus accumbens (NAcc), prefrontal cortex (Cg 3), and parietal cortex (Par 1). Animals were habituated for 3 days to a locomotor box, and after habituation, every second day for 5 weeks rats were placed into the locomotor chamber immediately after a subcutaneous injection of nicotine or saline. Rats demonstrated tolerance to an initial hypoactive response after each nicotine injection, and this was followed by an increase in activity after each injection (behavioral sensitization). This increase in activity was still present on a nicotine challenge after a 2-week abstinence period. One week after the nicotine challenge day, all rats were perfused and brains were removed. These brains we stained using Golgi-Cox procedures, and dendrites from the nucleus accumbens (N Acc), medial frontal cortex (Cg 3) and parietal cortex (Par 1) were analyzed using the camera lucida procedure. Results showed that rats receiving nicotine demonstrated an increase in dendritic length and spine density relative to controls in the NAcc and Cg3 brain areas, but not Par 1. The increase observed in the NAcc was significantly greater than what has been found with amphetamine or cocaine, and possible underlying mechanisms were discussed.