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Biomedical subjects

M Murphy

Publications and source records attributed to M Murphy.

At least 145 records · Page 8Linked to original sources
PubMed1999

Pax-3 regulates neurogenesis in neural crest-derived precursor cells.

The peripheral nervous system consists of multiple neural lineages derived from the neural crest (NC). Pax-3 is expressed in the NC and when mutated in the splotch mouse (Sp) results in the loss of derivatives from this precursor cell population. We have investigated the role of Pax-3 in regulating the generation of neurons from NC-derived precursor cells in vitro. Pax-3 mRNA in NC cultures is initially expressed in all NC but is subsequently only retained in neurons, suggesting a role in their generation. To determine whether Pax-3 is involved in neuron development, we first examined the generation of sensory-like neurons in NC cultures from Sp mice. Fivefold less sensory-like neurons were generated in NC cultures from Sp homozygous mice as compared to wild-type littermates. The role of Pax-3 in sensory neuron generation was then directly examined in dorsal root ganglia cultures by down-regulating the expression of Pax-3 protein with antisense oligonucleotides. It was found that antisense oligonucleotides inhibited 80-90% of newly generated sensory neurons; however, there was no significant effect on the survival of sensory neurons or the precursor population. These results suggest that Pax-3 has a role in regulating the differentiation of peripheral neurons.

Animals↗
PubMed1999

Gene expression and production of the monokine induced by IFN-gamma (MIG), IFN-inducible T cell alpha chemoattractant (I-TAC), and IFN-gamma-inducible protein-10 (IP-10) chemokines by human neutrophils.

Monokine induced by IFN-gamma (MIG), IFN-inducible T cell alpha chemoattractant (I-TAC), and IFN-gamma-inducible protein of 10 kDa (IP-10) are related members of the CXC chemokine subfamily that bind to a common receptor, CXCR3, and that are produced by different cell types in response to IFN-gamma. We have recently reported that human polymorphonuclear neutrophils (PMN) have the capacity to release IP-10. Herein, we show that PMN also have the ability to produce MIG and to express I-TAC mRNA in response to IFN-gamma in combination with either TNF-alpha or LPS. While IFN-gamma, alone or in association with agonists such as fMLP, IL-8, granulocyte (G)-CSF and granulocyte-macrophage (GM)-CSF, failed to influence MIG, IP-10, and I-TAC gene expression, IFN-alpha, in combination with TNF-alpha, LPS, or IL-1beta, resulted in a considerable induction of IP-10 release by neutrophils. Furthermore, IL-10 and IL-4 significantly suppressed the expression of MIG, IP-10, and I-TAC mRNA and the extracellular production of MIG and IP-10 in neutrophils stimulated with IFN-gamma plus either LPS or TNF-alpha. Finally, supernatants harvested from stimulated PMN induced migration and rapid integrin-dependent adhesion of CXCR3-expressing lymphocytes; these activities were significantly reduced by neutralizing anti-MIG and anti-IP-10 Abs, suggesting that they were mediated by MIG and IP-10 present in the supernatants. Since MIG, IP-10, and I-TAC are potent chemoattractants for NK cells and Th1 lymphocytes, the ability of neutrophils to produce these chemokines might contribute not only to the progression and evolution of the inflammatory response, but also to the regulation of the immune response.

Apoptosis↗
PubMed1999

Sonic hedgehog promotes neuronal differentiation of murine spinal cord precursors and collaborates with neurotrophin 3 to induce Islet-1.

Sonic hedgehog (Shh) is strongly implicated in the development of ventral structures in the nervous system. Addition of Sonic hedgehog protein to chick spinal cord explants induces floor plate and motoneuron development. Whether Shh acts directly to induce these cell types or whether their induction is mediated by additional factors is unknown. To further investigate the role of Shh in spinal neuron development, we have used low-density cultures of murine spinal cord precursor cells. Shh stimulated neuronal differentiation; however, it did not increase the proportion of neurons expressing the first postmitotic motoneuron marker Islet-1. Moreover, Shh did induce Islet-1 expression in neural tube explants, suggesting that it acts in combination with neural tube factors to induce motoneurons. Another factor implicated in motoneuron development is neurotrophin 3 (NT3), and when assayed in isolated precursor cultures, it had no effect on Islet-1 expression. However, the combination of N-terminal Shh and NT3 induced Islet-1 expression in the majority of neurons in low-density cultures of caudal intermediate neural plate. Furthermore, in explant cultures, Shh-mediated Islet-1 expression was blocked by an anti-NT3 antibody. Previous studies have shown expression of NT3 in the region of motoneuron differentiation and that spinal fusimotor neurons are lost in NT3 knock-out animals. Taken together, these findings suggest that Shh can act directly on spinal cord precursors to promote neuronal differentiation, but induction of Islet-1 expression is regulated by factors additional to Shh, including NT3.

Animals↗
PubMed1999

Suppression subtractive hybridization identifies high glucose levels as a stimulus for expression of connective tissue growth factor and other genes in human mesangial cells.

Accumulation of mesangial matrix is a pivotal event in the pathophysiology of diabetic nephropathy. The molecular triggers for matrix production are still being defined. Here, suppression subtractive hybridization identified 15 genes differentially induced when primary human mesangial cells are exposed to high glucose (30 mM versus 5 mM) in vitro. These genes included (a) known regulators of mesangial cell activation in diabetic nephropathy (fibronectin, caldesmon, thrombospondin, and plasminogen activator inhibitor-1), (b) novel genes, and (c) known genes whose induction by high glucose has not been reported. Prominent among the latter were genes encoding cytoskeleton-associated proteins and connective tissue growth factor (CTGF), a modulator of fibroblast matrix production. In parallel experiments, elevated CTGF mRNA levels were demonstrated in glomeruli of rats with streptozotocin-induced diabetic nephropathy. Mannitol provoked less mesangial cell CTGF expression in vitro than high glucose, excluding hyperosmolality as the key stimulus. The addition of recombinant CTGF to cultured mesangial cells enhanced expression of extracellular matrix proteins. High glucose stimulated expression of transforming growth factor beta1 (TGF-beta1), and addition of TGF-beta1 to mesangial cells triggered CTGF expression. CTGF expression induced by high glucose was partially suppressed by anti-TGF-beta1 antibody and by the protein kinase C inhibitor GF 109203X. Together, these data suggest that 1) high glucose stimulates mesangial CTGF expression by TGFbeta1-dependent and protein kinase C dependent pathways, and 2) CTGF may be a mediator of TGFbeta1-driven matrix production within a diabetic milieu.

Amino Acid Sequence↗
PubMed1999

Regulation of human IL-18 mRNA expression.

As virtually nothing is known about the pattern of expression of human IL-18, we investigated certain factors that may contribute to the regulation of IL-18 mRNA accumulation and compared this with regulation of the human gene encoding the p40 chain of IL-12, a cytokine that shares similar biologic activity with IL-18. IL-18 mRNA was expressed constitutively in unstimulated PBMC or monocytes, unlike p40, which required induction by a stimulus. Upon stimulation, IL-18 transcript accumulation was enhanced with an earlier and more transient pattern of expression than IL-12 p40 mRNA. Bacteria-derived stimuli and priming with IFN-gamma or IL-4 also upregulated IL-18 mRNA in a fashion similar to that of IL-12 p40. IL-10 exerted an inhibitory effect on IL-18 mRNA accumulation, though not as markedly as in the suppression of IL-12 p40 by IL-10. Finally, unlike IL-12 p40 mRNA, the constitutive accumulation of IL-18 transcripts by unstimulated cells was amplified in the presence of the translational blocker cycloheximide, which also caused a superinduction of IL-18 expression after Staphylococcus aureus stimulation.

Blotting, Northern↗
PubMed1999

Detection of clonal T-cell receptor gamma gene rearrangements in paraffin-embedded tissue by polymerase chain reaction and nonradioactive single-strand conformational polymorphism analysis.

The diagnosis of T-cell lymphoproliferative disorders, which frequently involve the skin and other extranodal sites, is often problematic because of the difficulty in establishing clonality in paraffin-embedded tissue. To this end, we developed a simple, nonradioactive method to detect T-cell receptor gamma (TCR-gamma) gene rearrangements by polymerase chain reaction single-strand conformational polymorphism (PCR-SSCP) in paraffin-embedded tissue. Jurkat and HSB-2 cell lines and peripheral blood samples from normal individuals were used as monoclonal and polyclonal controls, respectively. DNA was extracted from 24 biopsies of T-cell lymphomas, 12 biopsies of reactive lymphoid infiltrates, and 2 biopsies of primary cutaneous large B-cell lymphomas. Vgamma1-8, Vgamma9, Vgamma10, Vgamma11, and Jgamma1/Jgamma2 consensus primers were used for TCR-gamma gene rearrangement amplification and PCR products were analyzed by nonradioactive SSCP. Monoclonal controls yielded a well-defined banded pattern, whereas all polyclonal T-cell controls showed a reproducible pattern of smears. We detected monoclonality in 20/21 (95%) T-cell lymphoma cases, whereas no dominant T-cell clones were found in any of the reactive lymphoid infiltrates or B-cell lymphomas. Sensitivity of 1-5% was demonstrated by serially diluting Jurkat cells in mononuclear blood cells from normal individuals. We conclude that nonradioactive PCR-SSCP for TCR-gamma gene rearrangement analysis is a useful adjunct to routine histological and immunophenotypic methods in the diagnosis of T-cell lymphoproliferative disorders in paraffin-embedded tissue.

Base Sequence↗
PubMed1999

p27 expression in inflammatory bowel disease-associated neoplasia. Further evidence of a unique molecular pathogenesis.

The cyclin-dependent kinase inhibitor p27 is a negative regulator of the transition from G1 to S phase of the cell cycle, protects against inflammatory injury and promotes epithelial differentiation. Because p27 protein has been shown to be abnormally expressed both in dysplasia associated with Barrett's esophagus and in sporadic colorectal adenomas, we used immunohistochemistry to evaluate p27 expression in inflammatory bowel disease (IBD)-associated dysplasia and carcinomas. Normal, inflamed, and transitional mucosa, sporadic adenomas, and sporadic colonic carcinomas were studied as controls. In normal colonic epithelium p27 expression was restricted to the superficial, terminally differentiated cells. In colitic and inflamed diverticular mucosa p27 was expressed in the base of the crypts in 86 and 70% of cases, respectively. Similarly, in transitional mucosa adjacent to sporadic carcinomas p27 was expressed in the base of the crypts in all cases. Strong p27 expression extended more frequently from the base of the crypts to superficial cells in IBD-associated dysplasia than in sporadic adenomas (P < 0.007). Twenty of 20 (100%) IBD-associated carcinomas showed low p27 expression (<50% nuclei positive) compared to 6 of 20 (30%) stage-matched sporadic colorectal carcinomas (P < 0.001). We conclude (i) aberrant p27 protein expression in inflamed and IBD-associated nondysplastic mucosa is indistinguishable from that found in transitional mucosa adjacent to sporadic carcinomas; (ii) p27 is overexpressed in dysplastic lesions, perhaps as an attempt to counterbalance proliferative stimuli; and (iii) IBD-associated colorectal carcinomas have significantly lower p27 expression, commonly associated with poor prognosis, than stage-matched sporadic colorectal carcinomas. These findings further substantiate the existence of divergent molecular pathogenetic pathways between these types of carcinomas and suggest an intrinsically more aggressive behavior of IBD-associated colon carcinomas compared to sporadic ones.

Cell Cycle↗
PubMed1999

Detection of concurrent/recurrent non-Hodgkin's lymphoma in effusions by PCR.

A subset of patients with non-Hodgkin's lymphoma (NHL), present with or subsequently develop lymphocytic effusions. Differential diagnosis between reactive lymphocytosis and recurrent low-grade NHL is difficult by cytology alone. We studied the use of polymerase chain reaction (PCR)-based techniques to detect concurrent/recurrent NHL. Both primary tumors and atypical lymphocytic effusions of 12 low-grade B-NHL patients and 4 T-NHL patients were studied. Six pleural effusions (reactive/carcinomatous), in patients with no history of NHL, were included. Samples were amplified by PCR, using Fr3, Fr2, LJH, and VLJH primers specific for the immunoglobulin heavy chain (IgH) gene and Vgamma-8, Vgamma9, Vgamma10, Vgamma11 and Jgamma1/Jgamma2 consensus primers specific for the T-cell receptor gamma (TCR-gamma) gene. IgH gene PCR products were analyzed by polyacrylamide gel electrophoresis (PAGE). TCR-gamma gene PCR products were analyzed using a novel nonradioactive single-strand conformational polymorphism (SSCP) procedure. IgH gene rearrangement analysis demonstrated monoclonality in 11/12 primary low-grade B-NHLs. Identical monoclonal bands were found in both primary tumor and effusion in 9 patients. TCR-gamma gene rearrangement analysis demonstrated monoclonality in 4 of 4 primary T-NHLs. Identical monoclonal banded patterns were found in both primary tumor and effusion in 3 patients. Our results strongly support the diagnosis of concurrent/recurrent NHL in 13 of 16 (81%) cases of atypical lymphocytic effusions. IgH/PAGE and TCR-gamma/SSCP analyses are useful tools in the diagnoses of lymphocytic effusions in patients with NHL.

Diagnosis, Differential↗
PubMed1999

The intubating laryngeal mask: use of a new ventilating-intubating device in the emergency department.

The intubating laryngeal mask airway (ILM) was introduced in 1997 as a modification of the classic laryngeal mask airway. In addition to serving as an elective or emergency ventilating device, it is designed to allow blind intubation. We report 3 cases of airway management in the emergency department of Yale-New Haven Hospital where the ILM was used to establish ventilation and intubation in patients in whom direct laryngoscopy had failed. The 3 cases are representative of situations commonly seen in the ED: the obtunded and apneic ("crash airway") patient, failed rapid sequence intubation, and the recognized difficult airway/awake intubation. In all 3 cases, a clear airway was established on initial placement of the ILM, and intubation was achieved on the first attempt at blind advancement of the endotracheal tube. Although the ILM may be an important addition to the armamentarium of the emergency physician, proficiency in its use requires practice under controlled conditions. We suggest that the emergency physician seek out elective practice in either a teaching workshop or hospital operating theater.

Adult↗
PubMed1999

Ketamine in the ED: medical politics versus patient care.

The safe and effective use of ketamine for sedation/analgesia by emergency physicians has been validated in the medical literature. Nonetheless, arbitrary restrictions of this medication to anesthesia practitioners have prohibited emergency physician use in some locations. We explore the scientific evidence related to the use of ketamine by emergency physicians for sedation/analgesia, the history of sedation, the operational definitions of conscious sedation and dissociative anesthesia, and the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) related standards. We conclude that ketamine sedation/ analgesia offers many specific advantages for emergency patients and that it is safely administered by emergency physicians in the appropriately monitored setting.

Analgesics↗
PubMed1999

Dietary questions as determinants of mortality: the OXCHECK experience.

OBJECTIVE: To determine whether responses to simple dietary questions are associated with specific causes of death. DESIGN: Self-reported frequency intakes of various classes of foods and data on confounding factors were collected at the baseline survey. Death notifications up to 31 December 1997 were ascertained from the Office for National Statistics. Relative risk (RR) of death and 95% confidence intervals (CI) associated with baseline dietary factors were calculated by Cox regression. SETTING: Prospective follow-up study based on five UK general practices. SUBJECTS: Data were used from 11,090 men and women aged 35-64 years (81% of the eligible patient population) who responded to a postal questionnaire in 1989. RESULTS: After 9 years of follow-up, 598 deaths were recorded, 514 of these among the 10,522 subjects with no previous history of angina. All-cause mortality was positively associated with age, smoking and low social class, as expected. Among the dietary variables, all-cause mortality was significantly reduced in participants who reported relatively high consumption of vegetables, puddings, cakes, biscuits and sweets, fresh or frozen red meat (but not processed meat), among those who reported using polyunsaturated spreads and among moderate alcohol drinkers. These associations were broadly similar for deaths from ischaemic heart disease (IHD), cancer and all other causes combined, and were not greatly attenuated by adjusting for potential confounding factors including social class. CONCLUSIONS: Responses to simple questions about nutrition were associated with mortality. These findings must be interpreted with caution since residual confounding by dietary and lifestyle factors may underlie the associations.

Adult↗
PubMed1999

Subjective efficacy of nasal CPAP therapy in obstructive sleep apnoea syndrome: a prospective controlled study.

The response to nasal continuous positive airways pressure (nCPAP) of a wide variety of symptoms recognized to be associated with obstructive sleep apnoea syndrome (OSAS) was examined. Fifty-six consecutive patients with OSAS, confirmed by polysomnography (mean (SD) apnoea-hypopnoea index (AHI) 49.6 (22.6) events x h(-1), Epworth score 15.4 (5.0)), were asked to complete paired symptom evaluation questionnaires, before treatment and again after 4 months of nCPAP. The response rate was 80%. A control group of 21 consecutive OSAS patients of similar age, body mass index (BMI), AHI and Epworth score to the treated group but managed with conservative measures, completed the same questionnaires on two occasions, 4 months apart. The nCPAP-treated group showed significant reductions (Wilcoxon matched pairs test) in the symptoms of daytime sleepiness, restless sleep, heartburn, nocturia, enuresis, headache and nocturnal sweating, whereas controls showed no significant changes in these symptoms. There were no changes in BMI, smoking, alcohol consumption or exercise habits in either group. It was concluded that, in addition to improvements in symptoms of daytime sleepiness and restless sleep, a wide range of other symptoms may improve significantly with nasal continuous positive airways pressure therapy.

Case-Control Studies↗
PubMed1999

Genetic basis of transferase-deficient galactosaemia in Ireland and the population history of the Irish Travellers.

Transferase-deficient galactosaemia, resulting from deficient activity of galactose-1-phosphate uridyltransferase (GALT), is relatively common among the Travellers, an endogamous group of commercial/industrial nomads within the Irish population. This study has estimated the incidence of classical transferase-deficient galactosaemia in Ireland and determined the underlying GALT mutation spectrum in the Irish population and in the Traveller group. Based upon a survey of newborn screening records, the incidence of classical transferase-deficient galactosaemia was estimated to be 1 in 480 and 1 in 30,000 among the Traveller and non-Traveller communities respectively. Fifty-six classical galactosaemic patients were screened for mutation in the GALT locus by standard molecular methods. Q188R was the sole mutant allele among the Travellers and the majority mutant allele among the non-Travellers (89.1%). Of the five non-Q188R mutant alleles in the non-Traveller group, one was R333G and one F194L with three remaining uncharacterized. Anonymous population screening has shown the Q188R carrier frequency to be 0.092 or 1 in 11 among the Travellers as compared with 0.009 or 1 in 107 among the non-Travellers. The Q188R mutation was shown to be in linkage disequilibrium with a Sac I RFLP flanking exon 6 of the GALT gene. This represents the first molecular genetic description of classical transferase-deficient galactosaemia in Ireland and raises intriguing questions concerning the genetic history of the Irish Travellers.

Cohort Studies↗
PubMed1999

Psychological needs of patients with HIV disease: reviewing the literature using Nichols' (1985) Adjustment Reaction Model as a framework.

Since the first manifestations of HIV infection presented in the United Kingdom in the early 1980s, there has been a dramatic increase in the incidence of HIV infection and a proliferation of diseases associated with HIV infection. HIV infection is now associated with physical, psychological, mental and neurological conditions. Many of these conditions are serious and pose a major threat to physical and mental health, with rapid deterioration of health status necessitating multiple hospital admissions to acute hospitals. The majority of people hospitalized with HIV diseases are currently being cared for on general wards. While physical needs are often well met, psychological needs are to a certain extent neglected. The literature cites numerous cases of neglect, discrimination and threats to confidentiality by health professionals, including nurses. Such psychological needs, emanating out of hospitalization, compound existing psychological needs. Nichols' (1985) Adjustment Reaction Model describes a number of stages patients may go through during the course of their illness, and to some extent the patient's psychological needs can be traced throughout the trajectory of the disease. The purpose of this paper is to review the literature and present a theoretical argument, using Nichols' model as a framework, on psychological needs of patients with HIV diseases, and attempt to outline to what extent psychological needs of patients with HIV are met in institutional care. The authors recommend further research to determine nurses' perception of the importance of meeting psychological needs of patients with HIV disease.

Adaptation, Psychological↗