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D Murphy

Publications and source records attributed to D Murphy.

At least 55 records · Page 3Linked to original sources
PubMed2001

Species- and tissue-specific physiological regulation of vasopressin mRNA poly(A) tail length.

Transgenic experiments can be used to test the extent to which genes from different species can be swapped around, but still retain function, and be appropriately regulated. A vector has been developed that directs the expression of foreign genes to specific groups of vasopressin (VP) hypothalamic neurons in transgenic rats. Using this vector, we have expressed the bovine VP (bVP) RNA in the rat brain. In contrast to the situation in a mouse host, but like its endogenous rat counterpart, the mRNA encoded by the bVP transgene is subject to posttranscriptional physiological regulation in the hypothalamus; its poly(A) tail dramatically lengthens as a consequence of 3 days of dehydration. Transgene expression is also seen in the adrenal cortex, but here, despite a marked increase in transgene RNA levels with dehydration, there is no change in poly(A) tail length. These data suggest that the mouse hypothalamus and the rat adrenal gland do not have the transcript recognition or enzymatic machinery required for the physiologically responsive poly(A) tail length modulation seen in the rat brain.

Adrenal Cortex↗
PubMed2001

Estradiol induces formation of dendritic spines in hippocampal neurons: functional correlates.

Dissociated cultured rat hippocampal pyramidal neurons respond to estradiol with a time-dependent, twofold increase in density of their dendritic spines. This effect is mediated by an estrogen receptor, probably of the alpha nuclear receptor type. In searching for the molecular mechanisms leading from the initial activation of the estrogen receptor to the final formation of new dendritic spines, we found that estradiol acts on GABAergic interneurons expressing the estrogen receptor by decreasing their inhibitory tone. In culture, this is assumed to cause a shift in the balance between excitation and inhibition toward enhanced excitation, overactivation of the pyramidal neurons, and subsequent formation of novel dendritic spines. The action of estradiol on spine formation is mediated by phosphorylation of cyclic AMP response element binding protein in the pyramidal neurons and is blocked when inhibition is enhanced by diazepam and when excitation is blocked by tetrodotoxin. Progesterone blocks the effect of estradiol on dendritic spines through its conversion to tetrahydroprogesterone, which enhances GABAergic inhibition. Subsequent to formation of novel dendritic spines, there is an increase in the density of glutamatergic receptors in the affected cells, an increase in the cellular calcium response to glutamate, and an increase in network synaptic activity among the cultured neurons.

Animals↗
PubMed2001

In vivo assessment of the effects of estrogen on human brain.

There is increasing evidence from animal and in vitro studies to suggest that estrogen might have neuroprotective effects, and several plausible physiological mechanisms have been proposed. However, it is not yet fully understood how estrogen affects the human brain. There are several techniques that are currently employed for in vivo assessment of brain structure and function in humans, including neuropsychological and neuroendocrine testing, computerized tomography, structural and functional magnetic resonance imaging, magnetic resonance spectroscopy, single photon emission spectroscopy and positron emission tomography. Results from studies investigating the effects of estrogen on the female brain using the above techniques are reviewed here. The current data from humans suggest that the use of estrogen hormone-replacement therapy (HRT) in healthy, postmenopausal women might reduce the risk of developing Alzheimer's disease (AD) and preserve certain aspects of cognitive function. The use of HRT in postmenopausal women might also modulate neurotransmitter function and can increase cerebral blood flow in a regionally specific and task-dependent manner. In addition, the neuroprotective effects of HRT might depend on the length of its use. However, there is very little evidence at present that HRT is an effective treatment for established AD.

Brain↗
PubMed2001

Pain treatment satisfaction in spinal cord injury.

STUDY DESIGN: A survey on pain satisfaction was mailed to 300 individuals with spinal cord injury. Eighty-eight completed surveys were returned, and the results were analyzed. OBJECTIVES: The survey queried the respondents on characteristics of their pain, treatments received, the impact of pain on multiple, life activities and functions and the satisfaction with treatment received to reduce pain. SETTING: Subjects for the study were selected from the Spinal Cord Injury Registry from the Commonwealth of Virginia in the US. METHODS: Information was obtained from a survey sent to the subjects who were chosen randomly with respect to age and gender. At least 1 year had elapsed from the time of injury for each individual. RESULTS: Respondents were typically dissatisfied with the results of the treatments received to manage their pain. CONCLUSION: Pain in individuals with spinal cord injury needs to be addressed in a thorough fashion to reduce the adverse impact on life activities.

Adult↗
PubMed2001

HRT and its effect on normal ageing of the brain and dementia.

There are significant gender differences in human brain disease. For example, females are significantly more likely to suffer from Alzheimer's disease (AD) than men (even after correcting for differences in life expectancy), and females on hormone replacement therapy (HRT) are significantly less likely to suffer from Alzheimer's disease than women who do not take HRT. However the neurobiological basis to these differences in clinical brain disease were unknown until relatively recently. In this review we will discuss results of studies that show; (i) gender differences in human brain disease are most likely to be explained by gender differences in brain development and ageing; (ii) sex steroids have a significant effect on the brain; (iii) sex steroids are crucial to the development and ageing of brain regions affected in age-related brain diseases (for example AD); (iv) sex steroids interact with neuronal networks and chemical systems at many different levels; (v) sex steroids affect cognitive function in elderly women. Thus, the current literature supports the hypothesis that sex steroids can modulate brain ageing, and this provides a neurobiological explanation for the significantly higher prevalence of AD in females who do not take HRT, and may lead to new treatment approaches for age-related brain disease including AD.

Aging↗
PubMed2001

Unravelling mechanisms of action of angiotensin II on cardiorespiratory function using in vivo gene transfer.

We review recent and ongoing work from our laboratory that has shed novel insights into the effects of angiotensin II (ANGII) on the baroreflex at the level of the nucleus of the solitary tract (NTS). The NTS is the site of termination for baroreceptor afferents and is a potentially powerful region for neuronal modulation. ANGII applied to this nucleus attenuated the cardiac vagal and cardiac sympathetic components of the baroreceptor reflex. This effect was antagonized by blockade of either gamma-amino butyric acid receptors or nitric oxide synthase within the NTS. Interestingly, nitric oxide donors microinjected into the NTS mimicked the effect of ANGII. Using an adenovirus we showed that ANGII activated the endothelial isoform of nitric oxide synthase. The NTS was transfected to express a dominant negative truncated mutant form of endothelial nitric oxide synthase that prevented the depressant effect of ANGII on the baroreflex. Endothelial nitric oxide synthase was present in both neurones and endothelium in the NTS. A possibility is that ANGII activation of endothelial nitric oxide synthase is calcium dependent. However, in most NTS neurones tested, ANGII failed to elevate intracellular calcium concentration. We conclude that ANGII activates endothelial nitric oxide synthase to release nitric oxide which enhances gamma-amino butyric acid transmission destined for circuitry mediating the baroreflex. We discuss the contribution of endothelial cells within the nucleus of the solitary tract as a potential target for both circulating and/or centrally produced ANGII. These data have relevance to patients with essential hypertension and left heart failure, conditions in which ANGII activity is elevated and the baroreceptor reflex is depressed.

Adenoviridae↗
PubMed2001

Interferon-alpha delays S-phase progression in human hepatocellular carcinoma cells via inhibition of specific cyclin-dependent kinases.

The potential antiproliferative effects of interferon-alpha (IFN-alpha) in the treatment of hepatocellular carcinoma (HCC) are controversial, and the growth inhibitory mechanisms remain poorly understood. Therefore, the current study was designed to delineate the molecular mechanisms responsible for direct antiproliferative actions of IFN-alpha in HCC cells. IFN-alpha receptor expression and signal transduction were examined by RT-PCR, immunoprecipitation, Western analysis, and transient transactivation assays. Effects of IFN-alpha on cell growth and cell-cycle distribution were evaluated based on cell numbers and flow cytometry. Composition and activity of cyclin-dependent kinase complexes were determined by immunoblotting and histone-H1-kinase assays. Expression of IFN-alpha receptors was found in all 3 HCC cell lines. IFN-alpha binding initiated phosphorylation of Jak1 and Tyk2 kinases leading to Stat1/Stat2 activation, nuclear translocation, and transactivation of an ISRE-luciferase reporter gene construct. IFN-alpha treatment resulted in a time- and dose-dependent reduction of proliferation. Cell cycle analysis of G1-synchronized, IFN-alpha-treated HCC cells revealed a substantial delay in S-phase progression but no alteration of G1/S-phase transition or evidence of apoptotic cell death. Reflecting the time course of S-phase accumulation, cell cycle-dependent induction of Cyclin A and Cyclin B was impaired, resulting in reduced activity of Cdk2 and Cdc2 kinases. Furthermore, Cdc25C was selectively down-regulated. IFN-alpha treatment inhibits growth of HCC cells by specifically delaying S-phase progression, most likely because of inhibition of Cyclin A induction, resulting in decreased activity of the associated Cdk2 and Cdc2 kinases.

Antineoplastic Agents↗
PubMed2001

Surveillance of antibiotic resistance in invasive isolates of Neisseria meningitidis in Australia 1994-1999.

A total of 1434 strains of Neisseria meningitidis isolated from cases of invasive meningococcal disease (IMD) in Australia between 1994 and 1999 were examined by standard methods for susceptibility to antibiotics used for treatment and prophylaxis. The proportion of isolates fully susceptible to penicillin decreased from 45% in 1994 to 26% in 1999 (P<0.001). All the other isolates were less sensitive to penicillin except for two meningococci with a penicillin MIC of 1 mg/l. The geometric mean penicillin MIC increased from 0.045 to 0.065 mg/l from 1994 to 1999. There was no significant difference in the geometric mean penicillin MICs of serogroup B and serogroup C meningococci. Penicillin susceptibility was significantly associated with a poorer outcome. Isolates from survivors of IMD had a higher geometric mean penicillin MIC (0.06 mg/l) than those from fatal cases (0.048 mg/l) (P< 0.001). This suggests that factors other than the decrease in susceptibility to penicillin observed were more relevant to outcome in IMD. All isolates were fully susceptible to ceftriaxone. Rifampicin resistance was infrequent (eight isolates in 6 years) and sporadic. A single isolate had decreased quinolone susceptibility. Despite the significant shift in susceptibility to penicillin recorded, this group of antibiotics remains a suitable treatment for IMD in Australia.

4-Quinolones↗
PubMed2001

Postoperative changes in visual evoked potentials and cognitive function tests following sevoflurane anaesthesia.

We tested the hypothesis that minor disturbance of the visual pathway persists following general anaesthesia even when clinical discharge criteria are met. To test this, we measured visual evoked potentials (VEPs) in 13 ASA I or II patients who did not receive any pre-anaesthetic medication and underwent sevoflurane anaesthesia. VEPs were recorded on four occasions, before anaesthesia and at 30, 60, and 90 min after emergence from anaesthesia. Patients completed visual analogue scales (VAS) for sedation and anxiety, a Trieger Dot Test (TDT) and a Digit Symbol Substitution Test (DSST) immediately before each VEP recording. These results were compared using Student's t-test. P<0.05 was considered significant. VEP latency was prolonged (P<0.001) and amplitude diminished (P<0.05) at 30, 60, and 90 min after emergence from anaesthesia, when VAS scores for sedation and anxiety, TDT, and DSST had returned to pre-anaesthetic levels.

Adolescent↗
PubMed2001

Individual variation in the production of a 'bystander signal' following irradiation of primary cultures of normal human urothelium.

The existence of a bystander effect following both alpha and gamma irradiation of many cell lines is not now in dispute. The significance of this effect for cancer risk assessment and radiotherapy treatment planning requires demonstration of its relevance in vivo. The problem in demonstrating the existence of the effect in vivo is that other systemic effects may mask or confound the effect being investigated and it is practically impossible to attribute an effect in a particular cell to a signal produced in another irradiated cell. To approach this problem, we have developed an assay where fragments of human tissue can be irradiated ex vivo and the media harvested and added to unirradiated, allogenic explants or to a clonogenic cell line which has a well characterized and stable response to the bystander signal. The variation in production of the signal from patient to patient can thus be assessed using molecular and cellular endpoints. A study using tissue from over 100 patients and from mouse strains with well characterized responses to low level radiation exposure shows that there is variation in the effect of the signal produced by irradiated tissue from different patients. Gender, smoking status and the existence of a bladder malignancy influence the expression of the signal by normal urothelium. The effects of exposure to medium containing the signal are transmitted to distant progeny of the exposed cell population. The results may be important not only for understanding radiation risk mechanisms for protection but also for radiotherapy treatment planning where they may open new avenues for development of drugs for combined therapy.

Animals↗
PubMed2001

Gene regulation in the magnocellular hypothalamo-neurohypophysial system.

The hypothalamo-neurohypophysial system (HNS) is the major peptidergic neurosecretory system through which the brain controls peripheral physiology. The hormones vasopressin and oxytocin released from the HNS at the neurohypophysis serve homeostatic functions of water balance and reproduction. From a physiological viewpoint, the core question on the HNS has always been, "How is the rate of hormone production controlled?" Despite a clear description of the physiology, anatomy, cell biology, and biochemistry of the HNS gained over the last 100 years, this question has remained largely unanswered. However, recently, significant progress has been made through studies of gene identity and gene expression in the magnocellular neurons (MCNs) that constitute the HNS. These are keys to mechanisms and events that exist in the HNS. This review is an inventory of what we know about genes expressed in the HNS, about the regulation of their expression in response to physiological stimuli, and about their function. Genes relevant to the central question include receptors and signal transduction components that receive and process the message that the organism is in demand of a neurohypophysial hormone. The key players in gene regulatory events, the transcription factors, deserve special attention. They do not only control rates of hormone production at the level of the gene, but also determine the molecular make-up of the cell essential for appropriate development and physiological functioning. Finally, the HNS neurons are equipped with a machinery to produce and secrete hormones in a regulated manner. With the availability of several gene transfer approaches applicable to the HNS, it is anticipated that new insights will be obtained on how the HNS is able to respond to the physiological demands for its hormones.

Animals↗
PubMed2001

Production of transgenic rodents by the microinjection of cloned DNA into fertilized one-cell eggs.

Transgenic technologies that enable rapid movement between genotype and phenotype through specific loss-of-function, overexpression, or misexpression phenotypes will be crucial in the elucidation of gene sequences emerging from genome projects. This article describes detailed procedures for the generation of transgenic mice and rats by the injection of cloned DNA into the pronuclei of fertilized one-cell eggs.

Animal Husbandry↗
PubMed2001

Feasibility of a nurse-run asthma education program for urban African-Americans: a pilot study.

The objective of the study was to assess the feasibility of implementing and evaluating a culturally appropriate in-patient asthma education program specifically targeted for African-Americans. A consecutive sample of 28 African-American patients ages 18-50 who were hospitalized for asthma were randomized to an intervention group, which received three one-on-one sessions on chronic asthma management, or a control group, which received the usual care. Data on symptom frequency, self-management behaviors, quality of life, depression, and health care resource use were collected at baseline and at 3 and 6 months. Although the time required to recruit our sample took longer than anticipated, 28 subjects agreed to be in the study (70% acceptance rate) and complete the baseline interview. We observed no statistically significant differences from baseline or changing trends in frequency of asthma symptoms, self-management behaviors, and health care resource use between the intervention and control groups at 3 and 6 months. However patients in the intervention group demonstrated a greater average increase in asthma-related quality of life and a greater average decrease in depression than the control group. Feasibility issues included shortened length of stay, which necessitated conducting all three self-management sessions together; multiple interruptions during the sessions, and retention issues at 3- and 6-month follow-ups. The lessons learned from this pilot study are invaluable in that they will enable us to make changes in our existing protocol to ensure the success of a larger clinical trial.

Adolescent↗
PubMed2000

A novel function for the tumor suppressor p16(INK4a): induction of anoikis via upregulation of the alpha(5)beta(1) fibronectin receptor.

The tumor suppressor gene p16(INK4a) inhibits the kinase activity of the cyclin-dependent kinase 4-6/cyclin D complexes and subsequent phosphorylation of critical substrates necessary for transit through the G1 phase of the cell cycle. Recent studies suggested that control of the G1/S boundary might not be the sole biological function of p16(INK4a). We hypothesized that p16(INK4a) might influence hitherto unknown critical features of a malignant epithelial phenotype, such as anchorage dependence. Here we provide evidence that stable transfection of p16(INK4a) restitutes apoptosis induction upon loss of anchorage (anoikis) in a variety of human cancer cells. Anoikis in p16(INK4a)-transfected cells was evidenced by DNA fragmentation and poly(ADP-ribose) polymerase cleavage upon cultivation on polyhydroxyethylmethacrylate-coated dishes and was associated with suppression of anchorage-independent growth as well as complete loss of tumorigenicity. p16(INK4a)-mediated anoikis was due to selective transcriptional upregulation of the alpha(5) integrin chain of the alpha(5)beta(1) fibronectin receptor as detected by FACS((R)) analysis, immunoprecipitation, Northern blotting, and nuclear run-on assays. Addition of soluble fibronectin and inhibitory alpha(5) antibodies to nonadherent cells completely abolished p16(INK4a)-mediated anoikis, whereas laminin was ineffective. Furthermore, antisense-induced downregulation of the alpha(5) integrin chain in p16(INK4a)-transfected cells restored resistance to anoikis. These data suggest a novel functional interference between a cell cycle-regulating tumor suppressor gene and membrane-bound integrins, thus regulating a hallmark feature of an epithelial transformed phenotype: susceptibility to anoikis.

Animals↗