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Biomedical subjects

N Watson

Publications and source records attributed to N Watson.

At least 55 records · Page 3Linked to original sources
PubMed1998

RGS3 and RGS4 are GTPase activating proteins in the heart.

RGS family members are regulatory molecules that act as GTPase activating proteins (GAPs) for G alpha subunits of heterotrimeric G proteins. RGS proteins are able to deactivate G protein subunits of the Gi alpha, Go alpha and Gq alpha subtypes when tested in vitro and in vivo. Although the function of RGS proteins in cardiac physiology is unknown, their ability to deactivate Galpha subunits suggests that they may inhibit the action of muscarinic, alpha-adrenergic, endothelin, and other agonists. To evaluate the role of RGS family members in the regulation of cardiac physiology, we investigated the expression pattern of two RGS genes in normal and diseased rat heart tissue. RGS3 and RGS4 mRNAs and proteins were detected in adult myocardium. RGS3 and RGS4 gene expression was markedly enhanced in two model systems of cardiac hypertrophy: growth factor-stimulated cultured neonatal rat cardiomyocytes and pulmonary artery-banded (PAB) mice. RGS3 and RGS4 mRNA levels were reduced in failing myocardium obtained from SHHF/Mcc-fa(cp) (SHHF) rats. These findings support the hypothesis that RGS gene expression is highly regulated in myocardium and imply that RGS family members play an important role in the regulation of cardiac function.

Amino Acid Sequence↗
PubMed1998

The relevance of resting tension to responsiveness and inherent tone of human bronchial smooth muscle.

1. In the present study the effects of resting tension on isometric responses of human airway smooth muscle to contractile and relaxant stimuli were investigated. Also, its effects on inherent smooth muscle tone were examined, in an effort to determine a pre-defined resting tension which can be considered optimal for in vitro studies. 2. Bronchial ring preparations (2-4 mm internal diameter) were suspended in tissue baths at a range of levels of resting tension (200-1600 mg). The responses to electrical field stimulation (EFS, 30 V, 0.2 ms, 8-30 Hz), carbachol (3 microM), isoprenaline (1 microM) and the 5-lipoxygenase inhibitor, zileuton (10 microM) were investigated along with the ability of the preparations to recover stable resting tension after 60 min of washing post challenge. 3. EFS induced monophasic contractions at low stimulation frequency (8 Hz). However, with increasing frequency and tension (15 and 30 Hz; > 400 mg) contractile responses were accompanied by a relaxant component. The magnitude of contractile responses increased with increasing resting tension to a plateau at between 500 and 700 mg. Relaxant responses when present, increased in magnitude with increasing resting tension. 4. The level of resting tension did not significantly alter responses to carbachol, but tissues at tensions > or = 1000 mg showed poor tension recovery after washing. Isoprenaline-induced relaxations increased with increasing resting tension (P<0.05) and tension recovery after washing was complete. The 5-lipoxygenase sensitive portion of the tension (33+/-4%) was not altered by the level of resting tension. 5 These results suggest that in human bronchial ring preparations of 2-4 mm internal diameter, resting tensions within the range 400-1000 mg could be considered optimal for isometric tension recordings of protocols involving both contraction and relaxation procedures.

Adult↗
PubMed1998

A comparison of the cardiovascular effects of levobupivacaine and rac-bupivacaine following intravenous administration to healthy volunteers.

AIMS: The aim of this study was to compare the cardiovascular effects of levobupivacaine with those of rac-bupivacaine following i.v. administration to 14 healthy male volunteers. METHODS: Drugs were infused (at 10 mg min(-1)) using a randomized, double-blind, complete crossover procedure with a washout period of at least 1 week. The administration of drug was discontinued on the appearance of defined CNS symptoms or when a total of 150 mg had been given. Parameters measured were arterial blood pressure, heart rate, ECG, ejection fraction, acceleration index, stroke index and cardiac index. RESULTS: The mean doses administered were 56.1 mg and 47.9 mg for levobupivacaine and rac-bupivacaine respectively and the maximum mean plasma concentrations were 2.62 and 2.25 microg ml(-1) respectively. Despite the dose and plasma concentrations being comparable, levobupivacaine produced a statistically significant smaller reduction in mean stroke index (-5.14 vs -11.86 ml m(-2), P=0.001), acceleration index (-0.09 vs -0.20 s(-2), P=0.011) and the ejection fraction (-2.50 vs -4.29%, P=0.024). Both levobupivacaine (non significant) and rac-bupivacaine (significant) produced small increases in the PR interval and the corrected QT interval and although the effects of rac-bupivacaine appeared to be greater the difference between the two drugs was not significant. CONCLUSIONS: In conclusion, this study has shown that following i.v. administration levobupivacaine produces significantly less effects on cardiovascular function than does rac-bupivacaine. In particular the negative inotropic effect for levobupivacaine was less than that for rac-bupivacaine as indicated by changes in stroke index, acceleration index and ejection fraction.

Adult↗
PubMed1998

Histamine hypersensitivity induced by passive sensitization of human bronchus: effect of serum IgE depletion.

BACKGROUND: Sensitivity to specific allergens and increased sensitivity to common spasmogens are characteristic features of allergic asthma and are also features demonstrated by tissues passively sensitized with serum from atopic donors, displaying high levels of IgE. It is evident that the specific response to allergen is related to circulating levels of allergen-specific IgE, but it is unclear whether the histamine hypersensitivity is also related to this immunoglobulin. OBJECTIVE: The objective was to deplete IgE in the serum of a donor with high levels of total and allergen-specific IgE and compare specific-allergen sensitivity and sensitivity to histamine in tissues passively sensitized with either the whole serum or the IgE-depleted serum. METHODS: Serum from a Dermatophagoides farinae-sensitive asthmatic (total IgE = 1047 U/mL, D. farinae-specific IgE > 17.5 U/mL) was subjected to an immunomagnetic separation technique to reduce the levels of IgE (total and specific) to below 10 U/mL. Bronchial tissue from six non-atopic donors was then passively sensitized overnight with either the whole serum or IgE-depleted serum and D. farinae and histamine sensitivity were evaluated the next day using standard organ bath techniques. RESULTS: Passive sensitization with the whole serum resulted in the development of sensitivity to D. farinae and increased sensitivity to histamine (750+/-169 mg contraction to 10 U/mL D. farinae, histamine pEC50 5.64+/-0.16 and max. 813+/-109 mg in sensitized vs 37+/-34 mg contraction to 10 U/mL D. farinae histamine pEC50 5.05+/-0.23 and max. 490+/-84 in non-sensitized tissues, P>0.05). Incubation with IgE-depleted serum still produced histamine hypersensitivity (histamine pEC50 5.57+/-0.16 and max. 737+/-70 mg P>0.05), but no significant response to allergen was detected (20+/-13 mg contraction to 10 U/mL D. farinae). CONCLUSION: These results demonstrate, that increased reactivity to histamine and airway contraction to allergen induced by passive sensitization, occur through independent mechanisms and that, unlike allergen-sensitivity, histamine hypersensitivity is caused by a serum factor other than IgE.

Adolescent↗
PubMed1998

A case of monosomy 21 found to be an unbalanced de novo t(5p;21q) by fluorescence in situ hybridization.

A case is presented in which monosomy 21 was detected by routine cytogenetics and fluorescence in situ hybridization (FISH) studies demonstrated an unbalanced translocation t(5;21). The patient was partially monosomic for both 5p and 21q. The phenotype of the infant showed some features of the 5p- (cri-du-chat) syndrome, but there were also features present which were uncharacteristic of this syndrome. The present findings, combined with similar cases reported in the literature, provide further support for a proximal monosomy 21q syndrome.

Chromosome Aberrations↗
PubMed1998

RGS-GAIP, a GTPase-activating protein for Galphai heterotrimeric G proteins, is located on clathrin-coated vesicles.

RGS-GAIP (Galpha-interacting protein) is a member of the RGS (regulator of G protein signaling) family of proteins that functions to down-regulate Galphai/Galphaq-linked signaling. GAIP is a GAP or guanosine triphosphatase-activating protein that was initially discovered by virtue of its ability to bind to the heterotrimeric G protein Galphai3, which is found on both the plasma membrane (PM) and Golgi membranes. Previously, we demonstrated that, in contrast to most other GAPs, GAIP is membrane anchored and palmitoylated. In this work we used cell fractionation and immunocytochemistry to determine with what particular membranes GAIP is associated. In pituitary cells we found that GAIP fractionated with intracellular membranes, not the PM; by immunogold labeling GAIP was found on clathrin-coated buds or vesicles (CCVs) in the Golgi region. In rat liver GAIP was concentrated in vesicular carrier fractions; it was not found in either Golgi- or PM-enriched fractions. By immunogold labeling it was detected on clathrin-coated pits or CCVs located near the sinusoidal PM. These results suggest that GAIP may be associated with both TGN-derived and PM-derived CCVs. GAIP represents the first GAP found on CCVs or any other intracellular membranes. The presence of GAIP on CCVs suggests a model whereby a GAP is separated in space from its target G protein with the two coming into contact at the time of vesicle fusion.

Animals↗
PubMed1998

Inhibition of human airway sensitization by a novel monoclonal anti-IgE antibody, 17-9.

We investigated the effect of a novel mouse IgG2b nonanaphylactogenic anti-human IgE antibody, 17-9, on allergen and histamine responses in passively sensitized human airways in vitro to determine the specific contribution of IgE to the sensitization process. Bronchial rings were sensitized with serum containing high levels of allergen-specific IgE (Dermatophagoides farinae), or with a hapten-specific chimeric humanized IgE (JW8). There was a concentration-dependent contraction of serum-sensitized bronchial rings to D. farinae (517 +/- 188 mg tension at 10 U/ml, n = 8) that was not observed in nonsensitized controls. This response was practically abolished when tissues were sensitized in the presence of 100 microg/ml anti-IgE antibody 17-9 (54 +/- 20 mg). In tissues sensitized with the anti-NIP IgE, JW8, there was a concentration-dependent contraction to the specific antigen NIP-BSA (560 +/- 154 mg at 0.3 microg/ml, n = 5) that was not observed in nonsensitized control subjects and that was substantially inhibited when 17-9 was present in the sensitization buffer (124 +/- 109 mg). The inhibition with 17-9 was specific, as pretreatment with a non-IgE-specific IgG2b antibody did not affect allergen responses. Potency and maximal contractions to histamine in serum-sensitized tissues were significantly elevated compared with nonsensitized controls; this was not affected by the presence of 17-9 during sensitization (pEC50 = 5.1 +/- 0.2 versus 5.0 +/- 0.3 in tissues sensitized in the absence of 17-9). In tissues sensitized with JW8 there was no significant increase in responsiveness to histamine. We conclude that allergen responses in sensitized human airways are dependent on IgE levels in the sensitizing serum while nonspecific (hyper)responsiveness depends on serum factors other than IgE. Nonanaphylactogenic anti-human IgE antibodies effectively inhibit allergen responses of human airways in vitro but may not affect other factors inducing hyperresponsiveness.

Aged↗
PubMed1998

Laryngeal radionecrosis versus recurrent cancer: a clinical approach.

A group of 38 patients was studied by positron emission tomography utilizing fluorine18-labeled deoxyglucose (FDG-PET) scanning to examine the ability to differentiate postirradiation laryngeal recurrent cancer from radionecrosis. The resulting 79% correct diagnosis showed the technique to be far superior to conventional computed tomography scanning (61%) and clinical examination (43%). An algorithmic approach to evaluating and treating patients with radionecrosis and/or recurrent cancer by utilizing FDG-PET scanning is presented.

Algorithms↗
PubMed1997

RGS2/G0S8 is a selective inhibitor of Gqalpha function.

RGS (regulators of G protein signaling) proteins are GTPase activating proteins that inhibit signaling by heterotrimeric G proteins. All RGS proteins studied to date act on members of the Gialpha family, but not Gsalpha or G12alpha. RGS4 regulates Gialpha family members and Gqalpha. RGS2 (G0S8) is exceptional because the G proteins it regulates have not been identified. We report that RGS2 is a selective and potent inhibitor of Gqalpha function. RGS2 selectively binds Gqalpha, but not other Galpha proteins (Gi, Go, Gs, G12/13) in brain membranes; RGS4 binds Gqalpha and Gialpha family members. RGS2 binds purified recombinant Gqalpha, but not Goalpha, whereas RGS4 binds either. RGS2 does not stimulate the GTPase activities of Gsalpha or Gialpha family members, even at a protein concentration 3000-fold higher than is sufficient to observe effects of RGS4 on Gialpha family members. In contrast, RGS2 and RGS4 completely inhibit Gq-directed activation of phospholipase C in cell membranes. When reconstituted with phospholipid vesicles, RGS2 is 10-fold more potent than RGS4 in blocking Gqalpha-directed activation of phospholipase Cbeta1. These results identify a clear physiological role for RGS2, and describe the first example of an RGS protein that is a selective inhibitor of Gqalpha function.

Escherichia coli↗
PubMed1997

Inherent tone of human bronchus: role of eicosanoids and the epithelium.

1. Airway preparations of different species possess varying degrees of inherent tone which is the result of different metabolites of arachidonic acid in different species. In human bronchial smooth muscle in vitro we have investigated the effects of 5-lipoxygenase inhibition (zileuton, 10 microM), cyclo-oxygenase inhibition (indomethacin, 1 microM) and mechanical epithelium removal on inherent tone. The shunting of arachidonic acid by inhibition of one or other of these enzymes, as a possible explanation for the effects observed, has also been investigated. 2. Zileuton caused a significant fall in tone either alone (-107 +/- 33 mg) or after cyclo-oxygenase inhibition (-203 +/- 48 mg) and this effect was not significantly altered by epithelial removal (-191 +/- 43 mg alone; -333 +/- 88 mg after indomethacin). Indomethacin increased tone when applied alone (160 +/- 94 mg), but this effect only reached statistical significance after 5-lipoxygenase inhibition, (210+/-81 mg; P<0.05). Epithelial removal did not alter the effect of indomethacin when applied alone (213+/-97 mg), but significantly reduced the effect of indomethacin after 5-lipoxygenase inhibition (34 +/- 23 mg; P<0.05). 3. These data suggest that inherent tone in human bronchus is largely the result of contractile 5-lipoxygenase products. However, the involvement of cyclo-oxygenase products cannot entirely be discounted, since in the presence of 5-lipoxygenase inhibition contractile and relaxant eicosanoids originating from the bronchial epithelium appear to influence significantly inherent tone.

Adult↗
PubMed1997

Role of IgE in hyperresponsiveness induced by passive sensitization of human airways.

Incubation of airways from nonatopic patients with serum from patients with high IgE levels confers responsiveness to "specific" (allergen) and hyperresponsiveness to "nonspecific" (histamine) stimuli. We have tested the hypothesis that the level of IgE determines the degree of specific and nonspecific responsiveness. Bronchial rings from nonatopic patients were sensitized overnight with serum containing high levels of allergen-specific IgE, or with an allergen-specific chimeric IgE (JW8) in physiologic buffer. In vitro responsiveness to allergen and histamine was evaluated and compared with non-sensitized tissues from the same patients. Responses to specific allergen were demonstrated in all tissues sensitized with atopic serum or chimeric IgE, but not in nonsensitized tissues. Allergen responses were specific, since tissues sensitized using serum containing high Dermatohagoides farinae-specific IgE only, did not respond to either horse or dog allergens. The potency and magnitude of the maximal contraction to histamine was significantly (p < 0.05) increased in tissues sensitized using atopic serum with high total IgE concentrations compared with nonsensitized preparations, but was unchanged in tissues sensitized using chimeric IgE or serum with low total IgE levels. Therefore, specific IgE determines allergen responsiveness in passively sensitized human airways, but histamine hyperresponsiveness is independent of specific IgE and appears to be related to some other factor associated with serum containing high concentrations of total IgE.

Aged↗
PubMed1997

The local anaesthetic activity of levobupivacaine does not differ from racemic bupivacaine (Marcain): first clinical evidence.

The local anaesthetic efficacy of levobupivacaine was compared with racemic bupivacaine (Marcain) in healthy male volunteers who were undergoing ulnar nerve blockade. Levobupivacaine, like racemic bupivacaine, produced blockade of nerve function with evidence of a dose response relationship for levobupivacaine. There were no statistically significant differences with respect to duration of sensory pain, sensory touch or motor block when the adjusted mean for the levobupivacaine groups (0.125, 0.25 and 0.5%) were compared with the 0.25% racemic bupivacaine control group. It is concluded that levobupivacaine is an effective local anaesthetic in humans with a dose-related duration of effect. Its local anaesthetic effect did not differ from that of racemic bupivacaine.

Journal Article↗
PubMed1996

Two-way cell traffic between mother and fetus: biologic and clinical implications.

The bilateral trafficking of nucleated cells between the fetus and the mother was studied using polymerase chain reaction (PCR)-based systems sensitive enough to detect 1 target cell in 100,000 background cells. Sixty-six mother-baby pairs were recruited; maternal and cord blood samples were collected at delivery for DNA extraction. Cell trafficking was studied in informative cases using PCR-genotyping of polymorphic regions in the beta-globin cluster, the glutathione S-transferase M1 locus and the angiotensin converting enzyme gene. In addition, Y-PCR was also used in conjunction with these systems for the detection of fetal cells in maternal circulation. Fetal cells were detected in maternal peripheral blood in 26 of 51 cases whereas maternal cells were detected in 16 of 38 fetal umbilical cord blood samples. The proportion of umbilical samples with detectable maternal sequences was much higher than previously reported. In the 28 cases informative for both mother and baby, there was no obvious correlation between the cell traffic from mother to baby as compared to that from baby to mother. These findings may have implications for the use of cord blood for bone marrow transplantation, the vertical transmission of infectious agents, and the physiology of the feto-maternal relationship.

Blood Cells↗
PubMed1996

RGS family members: GTPase-activating proteins for heterotrimeric G-protein alpha-subunits.

Signaling pathways using heterotrimeric guanine-nucleotide-binding-proteins (G proteins) trigger physiological responses elicited by hormones, neurotransmitters and sensory stimuli. GTP binding activates G proteins by dissociating G alpha from G beta gamma subunits, and GTP hydrolysis by G alpha subunits deactivates G proteins by allowing heterotrimers to reform. However, deactivation of G-protein signalling pathways in vivo can occur 10- to 100-fold faster than the rate of GTP hydrolysis of G alpha subunits in vitro, suggesting that GTPase-activating proteins (GAPs) deactivate G alpha subunits. Here we report that RGS (for regulator of G-protein signalling) proteins are GAPs for G alpha subunits. RGS1, RGS4 and GAIP (for G alpha-interacting protein) bind specifically and tightly to G alphai and G alpha0 in cell membranes treated with GDP and AlF4(-), and are GAPs for G alphai, G alpha0 and transducin alpha-subunits, but not for G alphas. Thus, these RGS proteins are likely to regulate a subset of the G-protein signalling pathways in mammalian cells. Our results provide insight into the mechanisms that govern the duration and specificity of physiological responses elicited by G-protein-mediated signalling pathways.

Animals↗
PubMed1996

Isolated, electrically-stimulated airway preparations--their use in determining beta-adrenoceptor agonist activity.

We have assessed the suitability of electrically-stimulated superfused preparations of guinea-pig trachea, cat trachea and human bronchus for investigating the relaxant activity of the beta-adrenoceptor agonist, isoprenaline. Superfused strips of guinea-pig trachea, cat trachea and human bronchus all contracted in response to electrical stimulation. Guinea-pig trachea possesses inherent tone, and in its presence, electrical stimulation caused biphasic responses, comprising a modest, transient contraction, usually followed by a longer lasting relaxation. Human bronchus also possesses inherent tone, but responses were variable, generally monophasic, comprising a transient contraction of variable magnitude, but a longer lasting relaxation was occasionally observed after the transient contraction. Cat trachea possesses no inherent tone, and electrical stimulation of this preparation caused simple monophasic contractile responses. On guinea-pig trachea, addition of indomethacin (2.8 microM) abolished the inherent tone, and under these conditions, electrical stimulation caused monophasic contractile responses similar to those observed in cat trachea. On human bronchus, however, indomethacin enhanced inherent tone, which tended to uncover or exaggerate any relaxant component in the responses to electrical stimulation. The 5-lipoxygenase inhibitor, zileuton (10 microM), reduced, but did not abolish, the tone and converted the electrically-induced response to a monophasic contraction. In all preparations in which inherent tone was low or absent, whether naturally (cat trachea) or through pharmacological intervention (guinea-pig trachea with indomethacin, or human bronchus with zileuton), isoprenaline (1-100 nM) inhibited electrically-stimulated contractions in a concentration-related fashion (EC50s: 9-100 nM). In preparations exhibiting inherent tone (guinea-pig trachea with indomethacin or human bronchus with or without indomethacin), this tone was inhibited by isoprenaline. This relaxant activity, on guinea-pig trachea at least, was concentration-related (EC50: 5.4 nM). Such isoprenaline-induced relaxations complicated the analysis of inhibitory effects against electrically-induced contractions. Thus, in such experiments, only at higher concentrations did isoprenaline reliably inhibit these contractions (EC50: 23-119 nM), lower concentrations of isoprenaline often resulting in an apparent enhancement. The enhancement was probably artefactual, resulting from the fact that the electrically-induced contractions originated from a lower baseline. These data suggest that electrically-stimulated airway preparations are suitable for evaluating the relaxant activity of beta-adrenoceptor agonists, but the relaxant potency should be assessed in preparations lacking inherent tone, such as cat trachea, guinea-pig trachea in the presence of cyclo-oxygenase inhibition, or human bronchus in the presence of 5-lipoxygenase inhibition.

Adrenergic beta-2 Receptor Agonists↗
PubMed1996

The effect of temperature and growth rate on the susceptibility of Listeria monocytogenes to environmental stress conditions.

The effect of growth temperature and growth rate on the susceptibility to heat and pH stress were investigated in Listeria monocytogenes grown in continuous culture where these two growth variables could be varied independently of each other, and in batch culture. After growth at 30 degrees C or 10 degrees C at constant growth rate, or at 30 degrees C at different growth rates, cells did not differ in their resistance to heat at 55 degrees C. Cells grown at 30 degrees C were more resistant to acid stress at pH 2.5 than cells grown at the same growth rates at 10 degrees C. Cells grown at low growth rate at 30 degrees C gave greater resistance to acid stress than those grown at high growth rate. Growth temperature and growth rate had independent effects on the susceptibility of L. monocytogenes to acid stress conditions. This may have implications for the survival of L. monocytogenes in acidic foods.

Food Microbiology↗
PubMed1996

Selective muscarinic receptor agonists and antagonists.

Muscarinic receptors are composed of a family of four subtypes each of which can be distinguished pharmacologically and structurally. The physiological role of each subtype in the central and peripheral nervous systems remains to be clarified, due, in part, to a lack of agonists and antagonists with adequate subtype selectivity. Nonetheless, several agonists with functional selectivity for M1 receptors are now in advanced clinical evaluation for Alzheimer's disease, while selective M1/M3 antagonists may prove useful in the treatment of disorders of smooth muscle function. These novel compounds thus provide an advance over earlier therapeutics with which the clinical efficacy was compromised by the side effect profile. This review attempts to assess novel, selective agonists and antagonists, both in terms of their use in defining muscarinic receptor subtypes and their potential clinical utility.

Animals↗