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Biomedical subjects

S M Dunn

Publications and source records attributed to S M Dunn.

At least 73 records · Page 4Linked to original sources

Amyotrophic lateral sclerosis immunoglobulins are ineffective in altering calcium influx through presynaptic voltage-sensitive calcium channels.

Recent work has suggested that one factor in the etiology of the neuromuscular disease, amyotrophic lateral sclerosis (ALS), may be an autoimmune mechanism in which presynaptic voltage-sensitive calcium channels are an antigenic target. We have developed a fluorescence technique to measure rapid Ca2+ influx through presynaptic calcium channels in isolated nerve terminals (synaptosomes) from rat cerebral cortex. Depolarization of the synaptosomes by elevated external K+ concentration caused a rapid increase in cytoplasmic Ca2+, as measured by a change in fluorescence of the Ca2+ chelating dye, Fura-2, which was loaded inside the synaptosomes. Pharmacological characterization suggests that the P- and Q-subtypes of voltage-sensitive calcium channels mediate the majority of this Ca2+ influx. The synaptosome preparation has been used as a model system to investigate the effects of IgG, purified from eight ALS patients, on presynaptic calcium channel function. IgG (1 microgram ml-1 to 1 mg ml-1) was preincubated with the synaptosomes prior to depolarization. IgG, from these eight ALS patients, had no systematic effects on presynaptic Ca2+ influx. Thus, using this system, we find no evidence for an effect of ALS IgG on the function of presynaptic calcium channels.

Adult↗

Medicaid AIDS waivers: seeking cost effective financing of AIDS care.

The epidemic of Acquired Immunodeficiency Syndrome (AIDS) emerged at a time of transition in American health care and health care policy. The waiver of traditional Medicaid limitations on home and community-based health care services for persons with AIDS has been utilized by several states attempting to demonstrate models of care where quality of life as well as financial concerns are addressed. The Medicaid AIDS Waiver programs in New Jersey and Florida note promising results in the cost-effectiveness of such programs when compared with traditional Medicaid. Variations in patient participation by geographic area and demographic characteristics indicate areas for future improvement in program accessibility.

Acquired Immunodeficiency Syndrome↗

Processor quality assurance using digital imaging.

The purpose of this study was to develop and test a computer imaging method for assessing longitudinal processor variability. A technique called adaptive histogram equalization was used to test deviation in automatic processing when a test film is compared with a time-temperature processed standard at two different kVp's and film speeds. In a 14-day study we found that the density ranges for the standard film did not change, while the density range of the automatically processed films changed by a factor of two. These results suggest that if automatic processing is to be used for films taken as part of a longitudinal study-for example, subtraction-then processor variation can potentially lead to incorrect inference of bone gain or loss.

Absorptiometry, Photon↗

Molecular biology of the GABA(A) receptor: functional domains implicated by mutational analysis.

GABA(A) receptors are the major inhibitory neurotransmitter receptors in mammalian brain. They belong to a family of ligand-gated ion channels that also includes the nicotinic acetylcholine receptors, glycine receptors and 5HT(3) receptors. Each receptor in the family is believed to be a pentamer of homologous subunits that assemble to form a central transmembrane ion pore which, in the case of the GABA(A) receptor, is anion-selective. For almost twenty years, there has been tremendous interest in the structure and function of GABA(A) receptors, not only because of their importance in regulating brain excitability but also because these proteins are the specific targets for a wide variety of therapeutic agents including the anxiolytic benzodiazepines and barbiturates. Molecular cloning has revealed that GABA(A) receptors are heterogeneous, being formed by combinations of different isoforms of several subunit classes (alpha, beta, gamma, delta). The physiological and pharmacological properties of individual GABAA receptor subtypes appear to depend on their precise subunit complement. In this review, we focus on the application of modern techniques in molecular biology, particularly mutational analysis, to identify structural domains of these receptors that are important for ligand recognition and receptor function.

Amino Acid Sequence↗

When the diagnosis is cancer: patient communication experiences and preferences.

BACKGROUND: Discrepancies exist between reported experiences of patients when they have been given a diagnosis of cancer, published guidelines for telling a diagnosis, and patterns of communication patients rate as favorable. Several studies have identified what happened and what is important to cancer patients when told their diagnosis, but no studies have addressed subsequent communications concerning the implications of the diagnosis and treatment choices. This study extended previous research by investigating the experiences and preferences for communication about diagnosis, prognosis, and treatment of patients diagnosed with breast cancer or melanoma. METHODS: A self-report questionnaire was designed for this study based on previous research and qualitative data generated from focus groups. Patients with breast cancer or melanoma answered questions about their experiences with communication at the time of diagnosis and concerning prognosis, treatment and related issues. Comparisons were made between patient experiences, preferences and published guidelines. Differences between the experiences of breast cancer and melanoma patients were tested and the relationship between communication and subsequent psychological adjustment to cancer was assessed. RESULTS: Patient preferences for communication during diagnostic consultation were not always consistent with published guidelines. Type of cancer did not significantly affect patient preferences. Psychological adjustment was related to patient ratings of the quality of doctor discussion about treatment options, but not about the diagnosis of cancer and its implications. Patients who wanted more emotional support at the time of diagnosis subsequently experienced poorer psychological adjustment. CONCLUSIONS: The differences in patient preferences show that a list of prescriptions for how to disclose a cancer diagnosis is too simplistic. Guidelines for clinicians should be derived from patient-based data rather than be limited only to clinical opinion. Guidelines concerning communication at the time of diagnosis also need to address discussions concerning the implications of the diagnosis and making treatment decisions.

Adult↗

The major site of photoaffinity labeling of the gamma-aminobutyric acid type A receptor by [3H]flunitrazepam is histidine 102 of the alpha subunit.

The alpha subunit of the gamma-aminobutyric acid type A (GABA(A)) receptor is known to be photoaffinity labeled by the classical benzodiazepine agonist, [3H]flunitrazepam. To identify the specific site for [3H]flunitrazepam photoincorporation in the receptor subunit, we have subjected photoaffinity labeled GABA(A) receptors from bovine cerebral cortex to specific cleavage with cyanogen bromide and purified the resulting photolabeled peptides by immunoprecipitation with an anti-flunitrazepam polyclonal serum. A major photolabeled peptide component from reversed-phase high performance liquid chromatography of the immunopurified peptides was resolved by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The radioactivity profile indicated that the [3H]flunitrazepam photoaffinity label is covalently associated with a 5.4-kDa peptide. This peptide is glycosylated because treatment with the enzyme, peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase, reduced the molecular mass of the peptide to 3.2 kDa. Direct sequencing of the photolabeled peptide by automated Edman degradation showed that the radioactivity is released in the twelfth cycle. Based on the molecular mass of the peptides that can be generated by cyanogen bromide cleavage of the GABA(A) receptor alpha subunit and the potential sites for asparagine-linked glycosylation, the pattern of release of radioactivity during Edman degradation of the photolabeled peptide was mapped to the known amino acid sequence of the receptor subunit. The major site of photoincorporation by [3H]flunitrazepam on the GABA(A) receptor is shown to be alpha subunit residue His102 (numbering based on bovine alpha 1 sequence).

Affinity Labels↗

Mapping of GABAA receptor sites that are photoaffinity-labelled by [3H]flunitrazepam and [3H]Ro 15-4513.

The GABAA receptor in brain membranes prepared from bovine cerebral cortex and cerebellum has been photoaffinity-labelled by the classical benzodiazepine agonist, [3H]flunitrazepam, or by the partial inverse agonist [3H]Ro 15-4513. Following solubilization and precipitation with trichloroacetic acid, the photoaffinity-labelled receptor preparations were subjected to specific chemical cleavage using hydroxylamine, a reagent which cleaves specifically at a relatively rare Asn-Gly bond. The resulting peptides were resolved by denaturing polyacrylamide gel electrophoresis and mapping of these peptides to the known amino acid sequences of the GABAA receptor subunits has localized the photoaffinity-labelling sites for these two ligands to distinct portions of the alpha subunits. It is shown that the site for [3H]flunitrazepam photoaffinity-labelling in the receptor populations of both the cerebral cortex and cerebellum occurs within residues 1-103 of the bovine alpha 1 subunit sequence (or within analogous segments of homologous alpha subunits). In contrast, the site of photoaffinity-labelling by [3H]Ro 15-4513 in the cerebral cortex and in the diazepam-sensitive GABAA receptor population of the cerebellum lies between residues 104 and the carboxy-terminus of the bovine alpha 1 or homologous alpha subunits. However, the [3H]Ro 15-4513 photoaffinity-labelling site for the diazepam-insensitive receptors of the cerebellum is shown to occur within residues 1-101 (alpha 6 subunit numbering). These results demonstrate that the photoaffinity-labelling sites for [3H]flunitrazepam and [3H]Ro 15-4513 on the GABAA receptor are localized to distinct domains of the alpha 1 subunit and that [3H]Ro 15-4513 photoaffinity labels a site on the alpha 6 subunit that is unique from its site of labelling on the alpha 1 subunit.

Affinity Labels↗

Crystallization and preliminary diffraction studies of NodL, a rhizobial O-acetyl-transferase involved in the host-specific nodulation of legume roots.

The NodL specified O-acetyltransferase from the microbial symbiont Rhizobium leguminosarum has been over-expressed in Escherichia coli and purified using affinity-elution dye chromatography as the key step. The protein has been crystallized at 20 degrees C in 18% PEG 600, 0.1 M Tris/HCl buffer, pH 8.5, containing 1% dioxane, 0.25% octyl-beta-glucoside, and 5 mM coenzyme A using the hanging drop vapor diffusion method. Ambient temperature X-ray diffraction studies reveal the space group to be hexagonal (P6(3)22) with lattice constants a = b = 77.08 A, c = 160.6 A, and alpha = beta = 90 degrees, gamma = 120 degrees. Crystals that are flash-frozen to 120 K diffract beyond 2.7 A.

Acetyltransferases↗

Patient psychological and information needs when the diagnosis is diabetes.

People receiving a diabetes diagnosis must absorb information, change habits and adopt new behaviours almost immediately. Guidelines thoroughly cover the medical aspects of initial management however the information and psychological needs of patients are not addressed. This study aimed to identify diabetes patient needs for information and emotional support at the time of diagnosis. A sample of 1159 patients completed a questionnaire which assessed their concerns and information preferences at the time of diagnosis (response rate 64.4%). A sample of 100 general practitioners (GPs) were interviewed about their perceptions of these issues using a subset of items from the patient questionnaire. The diagnosis of diabetes was distressing for 60% of responders and 23% wanted more emotional support. Immediate issues such as injections were major concerns for patients and GPs correctly identified these. However GPs significantly overestimated complications as a concern for patients at diagnosis. Patients preferred diabetes educators and courses as sources of information. Satisfaction with information at diagnosis was high (80%), however younger patients were dissatisfied with their discussions concerning diabetes therapy. The findings suggest that clinicians giving a diagnosis of diabetes should: (1) be aware of patient variability in needs for emotional support and information preferences, (2) ask patients for their preferences and offer choices if available and (3) provide more information about treatment and increase patient involvement in discussions about therapy.

Adult↗

Identification of domains in human recombinant GABAA receptors that are photoaffinity labelled by [3H]flunitrazepam and [3H]Ro15-4513.

We have used [3H]flunitrazepam and [3H]Ro15-4513 as photoaffinity labelling agents in combination with a chemical cleavage technique to localize the benzodiazepine recognition sites of specific human recombinant alpha 1 beta 1 gamma 2, alpha 1 beta 3 gamma 2 and alpha 6 beta 3 gamma 2 GABAA receptor subtypes. The chemical agent utilized was hydroxylamine, whose substrate is a relatively rare asparagine-glycine amide bond that occurs only in the alpha subunits of the receptors examined in this study. Cleavage products were resolved using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The results of these experiments show that, in the alpha 1 subunit-containing receptors, incorporation of [3H]flunitrazepam occurs within residues 1-103 of the alpha 1 subunit, while incorporation of [3H]Ro15-4513 occurs within the region of the alpha 1 subunit that lies between residue 104 and the C-terminus. Photolabelling of membranes prepared from the alpha 6 beta 3 gamma 2-expressing cell line with [3H]Ro15-4513 resulted in the incorporation of radiolabel into two major protein species of M(r) 56,000 and M(r) 48,000, indicating incorporation into the alpha 6 subunit and possibly also the gamma 2 subunit. Hydroxylamine cleavage of alpha 6-containing receptors labelled with [3H]Ro15-4513 produced a gel profile consistent with the incorporation of the label occurring between residue 125 and the C-terminal. Thus, we have shown that the recognition sites for the agonist [3H]flunitrazepam and the inverse agonist [3H]Ro15-4513 occur within distinct domains of the human GABAA receptor.

Affinity Labels↗

On the receiving end. V: Patient perceptions of the side effects of cancer chemotherapy in 1993.

BACKGROUND: A study conducted in 1983 to identify and rank the symptoms experienced by patients receiving cancer chemotherapy reported that vomiting and nausea were the most important symptoms experienced. With the advent of new antiemetic regimens and changes in cancer chemotherapy, it was anticipated that changes may have occurred in patient perception of symptoms. The study was therefore repeated in 1993. PATIENTS AND METHODS: One hundred and fifty-five cancer patients receiving chemotherapy at a large urban teaching hospital participated in the study. Patients selected from cards listing symptoms all those experienced and the five most troublesome. RESULTS: Patients reported experiencing an average of 20 symptoms (13 physical and 7 psychosocial). Nausea was reported as the most severe symptom followed by tiredness and loss of hair. Vomiting which was the most severe symptom in 1983, now ranked 5th. Differences were detected in the symptoms experienced and reported as most severe, between chemotherapy regimens, between older and younger patients and between males and females. CONCLUSIONS: The results suggest a reduction in the severity of some symptoms experienced while receiving chemotherapy and a shift from concerns about physical to psychosocial issues.

Adult↗

A controlled study of anxiety and morbid cognitions at initial screening for human immunodeficiency virus (HIV) in a cohort of people with haemophilia.

AIM: This study examines the relationship between anxiety, psychological state and Human Immunodeficiency Virus (HIV) stages as defined by the Centers for Disease Control at the time of initial screening for HIV in a cohort of people with haemophilia who were at risk of prior exposure to HIV transmission from blood products. METHOD: Psychological scores, immunological measures, and clinical data from case notes for 116 potentially HIV exposed people with haemophilia attending initial screening for HIV infection in 1984-1985, were used to examine the relationship between psychological variables, clinical state and their clinical classification under the Centres for Disease Control categorization. Psychometric test results were obtained for 63 HIV seronegative patients and 53 HIV seropositive patients. Planned comparisons, multiple and logistic regressions, were used to explain observed differences between seronegative and seropositive subjects. The potential confounders of sex, age, severity of haemophilia, haemophilia type and blood product usage were controlled. RESULTS: The major finding of this study was that higher levels of State Anxiety at the time of initial screening for HIV, were observed in those patients who lacked recognized symptoms of HIV infection and were seropositive, compared with seronegative subjects. The State Anxiety scores were predicted by HIV infection or alternatively CD4+ T-cell levels. CONCLUSION: The findings of this study suggest that HIV infection can produce psychological effects prior to any physical symptoms of infection being apparent.

AIDS Serodiagnosis↗

Barriers and challenges in training health care providers for patient education.

Patient education and chronic illness are linked inextricably in the training of health care providers as we move to the year 2000. However, professional training remains organised around the traditional clinical interview with its focus on acute illness. Research into aspects of the health care consultation is well advanced, but we are only just beginning to understand the implications of the long-term interaction between patient and provider in the management of illnesses which cannot be cured. The challenges for health care providers are: to relinquish control of the consultation when it is appropriate; to be aware of the human biases affecting clinical judgment; to develop a professional approach to the human aspects of their relationships with patients and other professionals as part of a management team; and to focus on the longer term relationship and long-term benefit to the patient.

Female↗

Prediction of the location and time of spontaneous termination of reentrant ventricular tachycardia for radiofrequency catheter ablation therapy.

Ventricular tachycardia caused by reentrant excitation can lead to cardiac arrest and sudden death. Drug treatment and surgical procedures have been used with limited effectiveness. Catheter ablation methods are more promising because they are less invasive than surgery. Although ablation has come to be highly effective in the treatment of supraventricular tachycardias, the overall success rate remains low for ventricular tachycardias, which may be due in part to an inaccurate localization of the reentrant pathway. The authors hypothesize that a site in the myocardium exists that is critical for the maintenance or reentry and that when ablated, will result in permanent cessation of the tachycardia. The authors also hypothesize that this is the same site where the reentrant impulse blocks during spontaneous termination of tachycardia. A series of experiments has been designed to determine if there are specific properties of extracellular electrograms recorded from reentrant circuits that would enable the circuits to be identified without activation maps and, more specifically, allow the site of block causing spontaneous termination to be localized. For quantitative analysis of electrograms, a paradigm is developed to characterize electrogram morphology using a canine infarct model. Changes in morphology (shape, size, and location of signal deflections) can be considered (1) motions of a coordinate system and/or (2) conformational changes of shape. To a first approximation, stationarity over short time segments is assumed so that the motions and conformations can be parameterized. These parameters were extracted for 50 cardiac cycles during an episode of nonsustained ventricular tachycardia, in which 196-bipolar electrode pairs were positioned in an array format across the epicardial surface of the heart. The results of these studies of changes in electrogram morphology suggest that during cycles 5 to 49 of ventricular tachycardia, in many electrograms near the circuit, the cycle length increases linearly, the amplitude increases, and the duration of activation decreases. During cycles 50 to 54, the cycle length increases much more markedly, the amplitude decreases, and the duration of activation increases. These observations suggest that cycle lengthening may be an important property of some spontaneous terminations, and moreover that other morphologic characteristics are affected differently at different stages of cycle lengthening. Further, all motion parameters tended to oscillate from cycle to cycle in either an alternans pattern or longer oscillation. The variations in morphology were typically only a few percent from cycle to cycle. Such variability would not be evident using only ruler-and-caliper measurements made by hand because of the lack of precision and the sheer volume of data. It is expected that this approach for characterization of electrogram morphology will be extremely useful clinically to (1) increase speed and accuracy of ablation site selection and (2) reduce multichannel electrogram recording complexity during ablation site selection.

Algorithms↗

Computer-based interaction analysis of the cancer consultation.

There are few data available on which to base recommendations for effective communication in the cancer consultation. This paper describes a computerised interaction analysis system designed specifically for the cancer consultation and its application in a study investigating the relationship between doctor-patient behaviour and patient outcomes. One hundred and forty-two cancer patients attending their first consultation with a cancer specialist were audio taped and a copy of the tape was retained for interaction analysis. Before the consultation patient anxiety and information and involvement preferences were measured. Outcomes included recall of information, patient satisfaction with the consultation and psychological adjustment to cancer. Doctor behaviour was shown to vary significantly according to the age, sex, involvement preferences and in/out-patient status of the patient. The ratio of doctor to patient talk was related to satisfaction with communication, while patients whose questions were answered showed better psychological adjustment at follow-up. The results suggest that patient-centred consultations lead to improved satisfaction and psychological adjustment. These data provide precise information about consultation behaviour which can be used in the documentation of current practice and the evaluation of new interventions to improve communication.

Adolescent↗

Writing to referring doctors after a new patient consultation. What is wanted and what was contained in letters from one medical oncologist?

BACKGROUND: There are no Australasian data on the preferred information content of letters from consultant physicians to referring doctors. AIMS: To survey referring general practitioners (GPs) and specialists concerning their preferences for letters from a consultant physician after a new patient consultation to include different categories of information. To investigate the information content of letters written by one medical oncologist to referring doctors after a new patient consultation and contrast with that of individualised letters written to patients. METHODS: Fifty-five referring GPs and 53 specialists were sent a questionnaire seeking their views on the information content of letters from consultant physicians. Ninety-four letters after a new patient consultation were selected at random and analysed for their information content, and compared with 182 individualised letters sent to patients. RESULTS: Referring doctors wanted letters to contain details of diagnosis, clinical findings, test results and recommended future tests, treatment options, side effects and prognosis. Letters to referring doctors contained 19 items of information (range 8-33), while letters to patients contained a mean of 5.6 'salient' points (range 5-7). Both letters almost always stated the diagnosis, the presenting history and recommended treatment. Letters to patients more commonly presented information about prognosis, further tests, and explanation of symptoms than letters to doctors. In contrast, letters to referring doctors contained more information concerning the past medical, family, and drug history, clinical findings and test results. CONCLUSIONS: Letters sent by a consultant oncologist are not well tailored to the information needs of the referring clinician. Summary letters sent to patients may be modified to include information required by referring doctors.

Cross-Over Studies↗