Focal adhesion kinase (FAK), a non-receptor kinase, is a key regulator of integrin and focal adhesion signaling required for cancer cell survival, cell migration, and cell invasion. Amplification/Overexpression of FAK occurs in a wide variety of human cancers, supporting a role in carcinogenesis. Moreover, preclinical studies using cancer models where FAK is genetically inhibited indicate that this kinase is a potential therapeutic target to interfere with cancer progression. However, very little progress has been made in the identification of chemical inhibitors for potential therapeutic applications, in contrast to other kinases. Herein, I report optimization of the high-throughput in vitro Glo kinase assay for screening inhibitors of FAK kinase activity. Screening a large library of small molecule chemicals using these assays identified at least twenty FAK inhibitors, including a new FAK inhibitor developed by Pfizer and undergoing human clinical trials, and the non-specific kinase inhibitor staurosporine. Molecular studies of selective FAK inhibitors are undergoing in my host laboratory. In addition to this in vitro assay, I established similar assays to examine FAK kinase and adapter function in intact cells. The latter consists of ErbB-transformed cells deficient in FAK, and their matched cells where wild-type or kinase-dead FAK was restored. Biological characterization of these models revealed that both FAK kinase and adaptor activities cooperate for the regulation of cell migration, cell invasion, and tumor formation.

Bcr-abl expression being associated with anti-apoptotic signaling and expression of DNA repair enzymes, we surmised that single molecules capable of blocking abl tyrosine kinase (TK) function and damaging DNA should lead to compounds with potency superior to that of GleevecRTM. To this end, we designed novel agents termed "combi-molecules" programmed to not only behave as bcr-abl inhibitors on their own, but also to further degrade to another inhibitor and a DNA damaging species. The released inhibitor was designed to sustain bcr-abl inhibition following degradation of the combi-molecule and the DNA damaging species to activate pathways leading to apoptosis. To model this strategy termed "combi-targeting", we synthesized ZRCM5 (a monoalkyltriazene) that showed antiproliferative activity superior to that of the classical DNA damaging agent TemodalRTM, but not to that of Gleevec RTM. This result was imputed to the rather weak bcr-abl inhibitory activity of ZRCM5 and its strong DNA damaging property. Another prototype designed to contain an aniline mustard moiety (AK04) was a strong bcr-abl inhibitor but a poor DNA alkylating agent. Its cytotoxic activity was again stronger than that of the clinical alkylating agent chlorambucil but inferior to that of GleevecRTM. Further chemical studies directed at structural modification of the benzamide moiety led to the synthesis of ZRF1 with strong potency against bcr-abl TK and strong DNA damaging property. This novel optimized combi-molecule showed a 1.6-3-fold greater potency than GleevecRTM against bcr-abl expressing cells. Further investigation with ZRF1, showed that its cytotoxic potency was dependent on the p53 wild-type status of the cells. In cells expressing wild-type p53, p21 transactivation was associated with cell cycle arrest and that of Bax with apoptosis. In addition to, the pro-apoptotic effect of bcr-abl inhibition, these multiple mechanisms of action may synergistically enhance the cytotoxic potency of ZRF1 in p53 wild-type cells. The study conclusively demonstrated that p53 is a major determinant for the cytotoxic advantage of the novel combi-molecular approach in chronic myelogenous leukemia (CML), a disease in which 70-85% of all cases express wild-type p53.

Wernicke's encephalopathy is a neuropsychiatric disorder resulting from thiamine deficiency (TD) and is characterized by neuronal loss, astrocytic proliferation and microglial activation. Cyclooxygenases (COX) are enzymes which catalyze the first step in the synthesis of prostanoids. COX-1 is expressed constitutively and COX-2 is the inducible isoform. Groups of TD rats and pair-fed controls were killed at presymptomatic and symptomatic stages of encephalopathy. Cresyl violet and NeuN staining showed decreased numbers of neuronal cells in vulnerable regions (medial thalamus and inferior colliculus) but not in a spared region (frontal cortex). Numbers of GFAP-positive and OX-42-positive cells were increased at symptomatic stage of encephalopathy. Expression of COX-2 mRNA and neuronal COX-2 immunoreactivity were selectively increased in vulnerable regions of TD rats at symptomatic stages of encephalopathy. Nimesulide, a highly selective COX-2 inhibitor, lowered PGE2 levels and precipitated the progression of encephalopathy suggesting that COX-2 in this model is conferring neuroprotection.

Master of Pharmacy / Medicines are a fundamental healthcare intervention, but the benefits they provide depend entirely on the way in which they are used. This begins with prescribing, a complex task with substantial risks. Systematic evaluation of biomedical factors may be viewed as an essential component of this task, but prescribers also integrate an array of individual, social, cultural, environmental and commercial factors into their prescribing decisions. Furthermore, social and cultural characteristics of the prescriber’s workplace may influence how well prescribing decisions are carried out. Whilst numerous research efforts have helped to construct an in-depth understanding of non-biomedical influences on GP’s prescribing patterns, the characteristics of corresponding sorts of influences in teaching hospitals have not been well determined. In hospitals, supervised medical trainees, registrars and consultants prescribe within the framework of medicines management systems involving nurses, pharmacists and patients. Currently, little is known about whether each of these groups has distinct beliefs, attitudes and values that may affect either prescribing behaviour or how prescribing skills of medical trainees are acquired. The aim of this study was to explore the social and cultural dynamics of prescribing and prescriber training in teaching hospitals. To do this, established qualitative methods were employed. Junior doctors, registrars, consultants, nurses, and pharmacists from two metropolitan teaching hospitals were sampled purposively and invited to participate in semi-structured interviews. A brief questionnaire was used to collect demographic and contextual information. In the interviews, participants were asked about their attitudes towards prescribing, their perceptions of roles and responsibilities, how they communicated prescribing decisions, their perceptions of influences on prescribing, and their perceptions of factors contributing to prescribing errors. Participants were also asked for their opinions on various aspects of new prescriber training. Sampling proceeded until redundancy of themes was established. A pilot study was conducted with one participant from each professional group to optimise the interview schedule, and then using this tool, a further 38 participants were interviewed. In total, eight consultants, eight registrars, nine junior doctors, eleven pharmacists, and seven nurses participated. Using reiterative content analysis of a third of all transcripts, a coding scheme was developed, which was used to label and categorise the remaining transcripts. Categories were further developed and refined. The resultant core themes were cross indexed against the five different health professional types using thematic charts to explore patterns. The main lines of enquiry for this research were mapped, the properties of these categories and interrelationships explored in detail, and a model of the prescribing process was developed. Prescribing at the teaching hospitals was a complex process consisting of multiple steps undertaken by several different health professionals of varying levels of experience from three different health care disciplines. Because of the intricate separation of responsibilities, the operation of the process was highly reliant on the behaviours of each player and their relationships with each other. Key prescribing decisions associated with patient admissions were made, almost exclusively, by medical teams. Prescribing was therefore chiefly characterised by factors influencing the behaviours of the doctors. Their behaviours were influenced by factors relating to their individual characteristics (eg, knowledge, skills, experience); but also by a web of socio-cultural determinants inherent to the environment in which they worked. These factors were related to: the organisational structure of the prescribing process; the knowledge characteristics of the doctors; the communication patterns they used; the underlying assumptions they made about prescribing; and the work environment.

Master of Pharmacy / Medicines are a fundamental healthcare intervention, but the benefits they provide depend entirely on the way in which they are used. This begins with prescribing, a complex task with substantial risks. Systematic evaluation of biomedical factors may be viewed as an essential component of this task, but prescribers also integrate an array of individual, social, cultural, environmental and commercial factors into their prescribing decisions. Furthermore, social and cultural characteristics of the prescriber’s workplace may influence how well prescribing decisions are carried out. Whilst numerous research efforts have helped to construct an in-depth understanding of non-biomedical influences on GP’s prescribing patterns, the characteristics of corresponding sorts of influences in teaching hospitals have not been well determined. In hospitals, supervised medical trainees, registrars and consultants prescribe within the framework of medicines management systems involving nurses, pharmacists and patients. Currently, little is known about whether each of these groups has distinct beliefs, attitudes and values that may affect either prescribing behaviour or how prescribing skills of medical trainees are acquired. The aim of this study was to explore the social and cultural dynamics of prescribing and prescriber training in teaching hospitals. To do this, established qualitative methods were employed. Junior doctors, registrars, consultants, nurses, and pharmacists from two metropolitan teaching hospitals were sampled purposively and invited to participate in semi-structured interviews. A brief questionnaire was used to collect demographic and contextual information. In the interviews, participants were asked about their attitudes towards prescribing, their perceptions of roles and responsibilities, how they communicated prescribing decisions, their perceptions of influences on prescribing, and their perceptions of factors contributing to prescribing errors. Participants were also asked for their opinions on various aspects of new prescriber training. Sampling proceeded until redundancy of themes was established. A pilot study was conducted with one participant from each professional group to optimise the interview schedule, and then using this tool, a further 38 participants were interviewed. In total, eight consultants, eight registrars, nine junior doctors, eleven pharmacists, and seven nurses participated. Using reiterative content analysis of a third of all transcripts, a coding scheme was developed, which was used to label and categorise the remaining transcripts. Categories were further developed and refined. The resultant core themes were cross indexed against the five different health professional types using thematic charts to explore patterns. The main lines of enquiry for this research were mapped, the properties of these categories and interrelationships explored in detail, and a model of the prescribing process was developed. Prescribing at the teaching hospitals was a complex process consisting of multiple steps undertaken by several different health professionals of varying levels of experience from three different health care disciplines. Because of the intricate separation of responsibilities, the operation of the process was highly reliant on the behaviours of each player and their relationships with each other. Key prescribing decisions associated with patient admissions were made, almost exclusively, by medical teams. Prescribing was therefore chiefly characterised by factors influencing the behaviours of the doctors. Their behaviours were influenced by factors relating to their individual characteristics (eg, knowledge, skills, experience); but also by a web of socio-cultural determinants inherent to the environment in which they worked. These factors were related to: the organisational structure of the prescribing process; the knowledge characteristics of the doctors; the communication patterns they used; the underlying assumptions they made about prescribing; and the work environment.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 5 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2002. / Härtill 5 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv. / Härtill 6 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv. / Härtill 5 uppsatser.

Dissertation (Ph.D.)--University of South Florida, 2007. / Title from PDF of title page. Document formatted into pages; contains 187 pages. Includes vita. Includes bibliographical references.