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Biomedical subjects

T Sexton

Publications and source records attributed to T Sexton.

At least 19 recordsLinked to original sources
PubMed2005

Environmental reservoirs of methicillin-resistant Staphylococcus aureus in isolation rooms: correlation with patient isolates and implications for hospital hygiene.

Strategies to control and prevent the spread of methicillin-resistant Staphylococcus aureus (MRSA) include early identification of positive patients through screening, patient isolation, hand hygiene, nasal and skin decontamination, and the adequate cleaning and decontamination of clinical areas. However, many national and other guidelines provide few details on environmental decontamination regimens, partly because the role of the environment in the spread of MRSA is not well documented. We prospectively studied the environment of the isolation rooms of 25 MRSA patients for up to four weeks, sampling horizontal surfaces and the air using settle plates as well as an air sampler, while continuing regular daily cleaning according to the hospital protocol. We then typed 20 patient isolates and the corresponding environmental isolates (N=35) to assess the similarity of strains. A high proportion of samples were positive for MRSA; 269/502 (53.6%) surface samples, 70/250 (28%) air samples and 102/251 (40.6%) settle plates. Over half of the surface samples taken from the beds and the mattresses were positive for MRSA. Identical or closely related isolates were recovered from the patient and their environment in 14 (70%) patients, suggesting possible environmental contamination of the isolation rooms, possibly contributing to endemic MRSA. More effective and rigorous use of current approaches to cleaning and decontamination is required as well as consideration of newer technologies to eradicate MRSA and other hospital-acquired pathogens.

Air Microbiology↗
PubMed2004

Implications of colonization of vancomycin-resistant enterococci (VRE) in renal dialysis patients. Learning to live with it?

Vancomycin-resistant enterococci (VRE) commonly colonize, but less frequently infect, debilitated patients, such as those on chronic renal dialysis. The emergence of VRE amongst our cohort of renal replacement therapy patients posed considerable challenges in our attempts to prevent spread. Although 60 of 451 (13%) patients became colonized, only two patients required systemic antibiotics for confirmed or suspected invasive infection. Mortality and inpatient stay was greater in VRE-positive compared with VRE-negative patients (50% versus 10%) and patients who were screened on three or more occasions were likely to remain positive (e.g. 56% of patients screened on six occasions were positive). The application of recommended guidelines for the control of VRE, however, severely disrupted our renal dialysis programme and therefore had to be abandoned. As patients on renal dialysis are more likely to acquire VRE, remain colonized, require antibiotics and require regular inpatient or outpatient care more frequently than other patients, control measures should be adapted to minimize spread but not disrupt important and essential medical services.

Adolescent↗
PubMed2001

mu Opioid receptor-mediated G-protein activation by heroin metabolites: evidence for greater efficacy of 6-monoacetylmorphine compared with morphine.

The efficacy of heroin metabolites for the stimulation of mu opioid receptor-mediated G-protein activation was investigated using agonist-stimulated [(35)S]guanosine-5'-O-(gamma-thio)-triphosphate binding. In rat thalamic membranes, heroin and its primary metabolite, 6-monoacetylmorphine (6-MAM), were more efficacious than morphine or morphine-6-beta D-glucuronide. This increased efficacy was not due to increased action of heroin and 6-MAM at delta receptors, as determined by competitive antagonism by naloxone, lack of antagonism by naltrindole, and competitive partial antagonism with morphine. In agreement with this interpretation, the same relative efficacy profile of heroin and its metabolites was observed at the cloned human mu opioid receptor expressed in C6 glioma cells. Moreover, these efficacy differences were GDP-dependent in a manner consistent with accepted mechanisms of receptor-mediated G-protein activation. The activity of heroin was attributed to in vitro deacetylation to 6-MAM, as confirmed by HPLC analysis. These results indicate that the heroin metabolite 6-MAM possesses higher efficacy than other heroin metabolites at mu opioid receptors, which may contribute to the higher efficacy of heroin compared with morphine in certain behavioral paradigms in vivo.

Analgesics, Opioid↗
PubMed2001

Postnatal growth of intrinsic connections in mouse barrel cortex.

Surprisingly little is known about the development of connections within a functional area of the cerebral cortex. We examined the postnatal growth of connections in mouse barrel cortex during the second and third weeks after birth, coinciding with the period of rapid synaptogenesis that occurs just after the barrels first form. A barrel is a group of neurons in layer 4 of somatosensory cortex that is part of a cortical column. Each whisker/barrel column is linked anatomically and functionally to a homotopic whisker on the contralateral face. Radial groups of cortical neurons were labeled with the neuronal tracer biotinylated dextran amine in mice ranging in age from postnatal day 8 (P8; P0 is the date of birth) to adulthood. The spatial distributions of retrogradely labeled neurons in different laminae were analyzed. The barrel map in layer 4 was used as a template to compare quantitative data from different animals and to account for substantial changes in barrel and barrel field size during development. Intrinsic projections 1) innervate increasingly more distant targets within barrel cortex up to 3 weeks of age; 2) continue to form in targets after 3 weeks, effectively strengthening existing connections; 3) follow a timetable for growth that is layer-specific; 4) link more distant barrel columns in layer 4 from neurons that are found preferentially in the barrel side and the septa between barrels; and 5) form over the shortest distances between the barrel columns. These data indicate that intrinsic connections in mouse barrel cortex develop by the progressive addition of neuronal connections rather than by sculpting preliminary connections. We describe statistically significant changes in connectivity during development that may be applied to model and assess the development of connections after a variety of experimental perturbations, such as to the environment and/or the genome.

Aging↗
PubMed2001

Thermophilic biotrickling filtration of ethanol vapors.

The treatment of ethanol vapors in biotrickling filters for air pollution control was investigated. Two reactors were operated in parallel, one at ambient temperature (22 degrees C) and one at high temperature (53 degrees C). After a short adaptation phase, the removal of ethanol was similar in both reactors. At a bed contact time of 57 s, the elimination capacity exceeded 220 g m(-3) h(-1) at both temperatures. The experiments performed revealed that the process was most likely limited by biodegradation in the biofilm. The high-temperature biotrickling filter exhibited a higher degree of ethanol mineralization to CO2 (60 vs 46% at ambient temperature); hence, a lower rate of biomass accumulation was observed. Plating and cultivation of biofilm samples revealed that the high-temperature biotrickling filter hosted a process culture composed of both mesophilic and thermotolerant or thermophilic microorganisms, whereas the ambient-temperature reactor lacked microorganisms capable of growing at high temperature. Consequently, the performance of the control biotrickling filter was significantly affected by a short incursion at 53 degrees C. The upper temperature limit for treatment was 62 degrees C. Overall, the results of this study open new possibilities for biotrickling filtration of hot gases.

Air Pollution↗
PubMed2001

Synthesis of 2beta-acyl-3beta-(substituted naphthyl)-8-azabicyclo[3.2.1]octanes and their binding affinities at dopamine and serotonin transport sites.

A series of 3beta-naphthyltropane derivatives were synthesized and found to show high affinity at both the dopamine and serotonin transporter sites, leading to some of the most potent inhibitors known based on the tropane structure. Comparative molecular field analysis (CoMFA) models were developed for both dopamine and serotonin transporter binding data. These models provide insights into those factors that influence binding at the two transporters.

Animals↗
PubMed2000

Characterization of a tropane radioligand, [(3)H]2beta-propanoyl-3beta-(4-tolyl) tropane ([(3)H]PTT), for dopamine transport sites in rat brain.

PTT (2beta-propanoyl-3beta-[4-tolyl] tropane) is a tropane analog relatively selective for dopamine transporters in binding and uptake assays in vitro, with long-acting psychostimulant properties in vivo. To explore its utility in binding to dopamine transporters, [(3)H]PTT was synthesized and assayed for binding in rat striatal membranes and by in vitro autoradiography. In membranes, binding of [(3)H]PTT was saturable to a single class of binding sites with a K(D) value of 3 nM. The pharmacology of [(3)H]PTT binding in striatal membranes was consistent with that of a ligand selective for dopamine transporters, with dopamine-selective compounds being significantly more potent in displacing [(3)H]PTT binding than those for 5-HT or norepinephrine transporters. Although the ability of various transporter inhibitors to displace both [(125)I]RTI-55 and [(3)H]PTT binding correlated significantly with each other, there was a better correlation of inhibitor potencies versus [(3)H]PTT binding and dopamine uptake than versus [(125)I]RTI-55 binding and dopamine uptake. The differences in correlations were most noticeable for compounds relatively selective at the 5-hydroxytryptamine (serotonin) transporter. The autoradiographic distribution of [(3)H]PTT binding in coronal sections was consistent with the known distribution of the dopamine transporter, with high levels of binding evident in caudate nucleus, nucleus accumbens, and olfactory tubercle. Moderate densities of [(3)H]PTT binding were also observed in substantia nigra pars compacta, and ventral tegmental area, as well as in the anterior cingulate cortex and portions of the hypothalamus. In addition, nonspecific binding was less than 5% of total binding. Thus, [(3)H]PTT provides an accurate and convenient marker for the dopamine transporter.

Animals↗
PubMed1999

Regulation of rat dopamine transporter mRNA and protein by chronic cocaine administration.

This study describes a direct comparison of dopamine transporter (DAT) mRNA and protein, as well as its binding sites, in tissue from the same animals after chronic cocaine administration. Rats were treated twice daily with 25 mg/kg cocaine or with saline. After 8 days of cocaine administration, changes in DAT mRNA levels in the substantia nigra pars compacta and ventral tegmental area were measured by in situ hybridization, and DAT protein in the striatum was quantified by immunoblotting. Whereas chronic cocaine treatment significantly reduced levels of DAT mRNA in the substantia nigra pars compacta and ventral tegmental area as compared with vehicle-treated controls, cocaine treatment did not alter DAT protein levels in the striatum. Furthermore, the density of DAT binding sites was also measured in the striatum by quantitative autoradiography using two DAT radioligands, 33-(4-[125I]iodophenyl)tropane-2-carboxylic acid methyl ester ([125I]RTI-55) and [3H]propanoyl-3beta-(4-tolyl)tropane ([3H]PTT). Similar to the results of immunoblotting of DAT protein, [1251]RTI-55 and [3H]PTT binding site levels also remained unaltered. These results indicate a dissociation in the regulation of DAT mRNA and its protein levels as a result of cocaine administration in rats. This study also indicates that the DAT ligands [3H]PTT and [125I]RTI-55 provide an accurate assessment of DAT protein levels.

Animals↗
PubMed1998

Long-acting blockade of biogenic amine transporters in rat brain by administration of the potent novel tropane 2beta-propanoyl-3beta-(2-Naphthyl)-tropane.

2beta-Propanoyl-3beta-(2-naphthyl)-tropane (WF-23) is a potent cocaine analog with activity at dopamine and serotonin transporters. The purpose of these experiments was to characterize the time course of effects of acute administration of WF-23 on spontaneous locomotion and biogenic amine transporters. Rats received injections i.p. with WF-23 (1 mg/kg), cocaine (30 mg/kg) or vehicle and locomotor activity was measured at various times postinjection. Animals were killed immediately after behavioral activity. Locomotor activity was significantly increased by WF-23 administration, reaching maximum at 4 hr and persisting for 24 hr. Cocaine-elicited elevations in locomotor activity occurred only at the earliest times. WF-23 decreased DA transporter binding in striatal membranes ([125I]RTI-55 binding), with >50% loss in binding for up to 49 hr postinjection. WF-23 increased the Kd of the high affinity site, with no effect on Bmax. Cocaine depressed binding (20%) only at the earliest times. WF-23 decreased levels of [3H]WIN 35,428 binding sites up to 95% of control in both dorsal and ventral striatum with a similar time-course when assessed autoradiographically. WF-23 also reduced [3H]citalopram binding to serotonin transporter sites throughout the brain. The slow onset and very long duration of action of WF-23, taken together with its actions at dopamine and serotonin transporters, suggest a potential role for treatment of disorders characterized by their involvement of these neural systems.

Animals↗
PubMed1996

Synthesis of 3 beta-aryl-8-azabicyclo[3.2.1]octanes with high binding affinities and selectivities for the serotonin transporter site.

A novel entry to tropane analogs of cocaine was developed based on the reaction of rhodium-stabilized vinylcarbenoids with pyrroles. These analogs were tested in binding to dopamine, serotonin (5-HT), and norepinephrine transporters in membranes from rat striatum and frontal cortex. In all the analogs, the aryl group at the 3 position was directly bound to the tropane ring and an ethyl ketone moiety was present at the 2 position. By appropriate modification of the aryl and nitrogen substituents, highly potent and 5-HT selective tropanes were prepared. The most potent and selective compound was 3 beta-[4-(1-methylethenyl)phenyl]-2 beta-propanoyl-8-azabicyclo[3.2.1]octane (13b) which had a Ki of 0.1 nM at 5-HT transporters and was 150 times more potent at 5-HT vs dopamine transporters and almost 1000 times more potent at 5-HT vs norepinephrine transporters.

Animals↗
PubMed1995

Novel 2-substituted cocaine analogs: uptake and ligand binding studies at dopamine, serotonin and norepinephrine transport sites in the rat brain.

A novel scheme utilizing vinylcarbenoid precursors has been developed for the synthesis of tropane analogs of cocaine. Specificities of these compounds for dopamine (DA), serotonin (5-HT) or norepinephrine (NE) transporters were determined by both uptake inhibition and binding assays. In each of the analogs, the aryl group at position 3 was bound directly to the tropane ring (as in WIN-35,428), and methyl or ethyl ketone moieties were present at position 2 in lieu of the ester linkage present in cocaine. The addition of methyl or ethyl ketone groups in position 2 did not affect potency compared to ester groups in the same position, but substituents on the benzene ring greatly affected potencies. The analogs could be categorized according to their relative specificity, which consisted of those selective for DA and NE transporters with little affinity for the 5-HT transporter (e.g., WF-39), those selective for only the DA transporter (e.g., WF-29) and those selective for the 5-HT transporter (e.g., WF-31) with much less affinity for the DA and NE transporters. There also were those analogs (e.g., WF-23) with high affinity, but with equal affinity at all three transporters. The analogs displayed significant correlation between uptake inhibition and binding displacement at DA, 5-HT and NE transport sites. These results suggest that large variations in the tropane structure do not result in a differentiation between binding at biogenic amine transporters and inhibition of amine uptake.

Adrenergic Uptake Inhibitors↗
PubMed1994

Synthesis of 2 beta-acyl-3 beta-aryl-8-azabicyclo[3.2.1]octanes and their binding affinities at dopamine and serotonin transport sites in rat striatum and frontal cortex.

A novel entry to tropane analogs of cocaine was developed on the basis of the reaction of rhodium-stabilized vinylcarbenoids with pyrroles. These analogs were tested in binding to dopamine and serotonin (5-HT) transporters in membranes from rat striatum and frontal cortex. In all the analogs, the aryl group at the 3-position was directly bound to the tropane ring (as in WIN-35,428), and methyl or ethyl ketone moieties were present at the 2-position instead of the typical ester group. The series of analogs containing a 2-naphthyl group at the 3-position were most potent, with Ki values < 1 nM in binding to both dopamine and 5-HT transporters. Although the unsubstituted 2-naphthyl analog was nonselective at dopamine and 5-HT transport sites, other compounds were selective for either site. In general, compounds with relatively small substituents on the aromatic moiety (such as p-methyl or p-fluoro) were relatively selective for the dopamine transporters, while a p-isopropylphenyl derivative was selective for the 5-HT transport sites. This latter compound represents the first N-methyltropane derivative specific for 5-HT transporters. Resolution of two of the most significant analogs was achieved by HPLC on a chiral stationary phase; the active enantiomer of a 2-naphthyl analog exhibited Ki values of < 0.1 nM at both dopamine and 5-HT transporter sites.

Animals↗
PubMed1994

Effects of anandamide on cannabinoid receptors in rat brain membranes.

Anandamide (arachidonylethanolamide) is a compound recently isolated from porcine brain as a putative endogenous ligand at cannabinoid receptors. The present studies examined the effects of anandamide on cannabinoid receptor binding sites and adenylyl cyclase in rat brain membranes. Receptor binding experiments, conducted at 25 degrees for 90 min, apparently resulted in significant degradation of anandamide, since anandamide (10 microM) had little effect on [3H]WIN 55212-2 binding in cerebellar membranes. Addition of the general serine protease inhibitor phenylmethylsulfonyl fluoride (PMSF) protected against this degradation, resulting in an IC50 value of 90 nM for anandamide versus [3H]WIN 55212-2 binding. Anandamide inhibited adenylyl cyclase in cerebellar membranes in a GTP-dependent manner, exhibiting a maximal inhibition level slightly less than that of WIN 55212-2 and CP-55,940, with an IC50 value of 1.9 microM. The effect of anandamide on adenylyl cyclase was region-specific, with maximal inhibition occurring in cerebellum and striatum. These results suggest that anandamide acts at G-protein-coupled cannabinoid receptors in brain with properties similar to those of exogenous cannabinoids.

Adenylyl Cyclase Inhibitors↗
PubMed1993

Novel 2-substituted cocaine analogs: binding properties at dopamine transport sites in rat striatum.

A novel scheme utilizing vinylcarbenoid precursors has been developed for the synthesis of novel tropane analogs of cocaine. Using this method, 15 analogs were prepared and tested for activity in binding to dopamine transporters in rat striatal membranes using [125I]RTI-55. In all the analogs, the aryl group at the 3 position was directly bound to the tropane ring (as in WIN 35,428), and methyl or ethyl ketone moieties were present at the 2 position instead of the typical ester group. The most potent analog was a 2-naphthyl derivative (IC50 value of 0.2 nM, vs. 170 nM for cocaine), while replacement of the aryl with either ethyl or cyclohexyl drastically reduced potency (to > 50 microM and 5 microM, respectively).

Animals↗
PubMed1993

Biochemical and genetic basis for the ETS (enterotoxin sensitivity) phenotype in mice.

Evidence has been accumulating that humans are genetically predisposed to cholera gravis. Using the sealed adult mouse model, we found that certain inbred mice were also hypersensitive to cholera toxin (CT). Such mice were designated ETS+ (enterotoxin sensitive), and the trait was linked to the K end of the mouse H-2 histocompatibility complex. Cells isolated from ETS+ mice bound more CT and accumulated more cyclic adenosine monophosphate (cAMP) after intoxication. Analysis of ETS+ cells showed that they express lesser amounts of the non-GM1 gangliosides that block or compete for relevant CT binding sites in ETS- cells. Conversion of ETS- non-GM1 gangliosides to GM1 with neuraminidase increased CT binding and cAMP responses. Reconstitution of nonreactive ganglioside-deficient cells with ETS+ or ETS- gangliosides caused them to bind CT like the original ETS+ or ETS- cells. Ganglioside expression genes known to map to the same H-2-linked region as the ETS phenotype seem to be involved in controlling murine susceptibility to CT.

Animals↗
PubMed1992

Opioid and cannabinoid receptor inhibition of adenylyl cyclase in brain.

Both opioids and cannabinoids bind to G-protein-coupled receptors to inhibit adenylyl cyclase in neurons. These reactions were assayed in brain membranes, where maximal inhibitory activity occurred in the following regions: mu-opioid inhibition in rat thalamus, delta-opioid inhibition in rat striatum, kappa-opioid inhibition in guinea pig cerebellum, and cannabinoid inhibition in cerebellum. The inhibition of adenylyl cyclase by both cannabinoid and opioid agonists was typical of G-protein-linked receptors: they required GTP, they were not supported by non-hydrolyzable GTP analogs, and they were abolished (in primary neuronal cell culture) by pertussis toxin treatment. The immediate targets of this system were determined by assaying protein phosphorylation in the presence of receptor agonists and App(NH)p, a substrate for adenylyl cyclase. In striatal membranes, opioid agonists inhibited the phosphorylation of at least two bands of MW 85 and 63 kDa, which may be synapsins I and II, respectively. Other experiments determined the long-term effects of this second messenger system. In primary neuronal cultures, opioid-inhibited adenylyl cyclase attenuated forskolin-stimulated pro-enkephalin mRNA levels, thus providing a feedback regulation of opioid synthesis. Finally, in cerebellar granule cells, both cannabinoid and opioid receptors may exist on the same cells. In these cells, agonists which bind to different receptor types may produce similar biological responses.

Adenylyl Cyclase Inhibitors↗
PubMed1992

Opioid-inhibited adenylyl cyclase in rat brain membranes: lack of correlation with high-affinity opioid receptor binding sites.

Opioid agonists bind to GTP-binding (G-protein)-coupled receptors to inhibit adenylyl cyclase. To explore the relationship between opioid receptor binding sites and opioid-inhibited adenylyl cyclase, membranes from rat striatum were incubated with agents that block opioid receptor binding. These agents included irreversible opioid agonists (oxymorphone-p-nitrophenylhydrazone), irreversible antagonists [naloxonazine, beta-funaltrexamine, and beta-chlornaltrexamine (beta-CNA)], and phospholipase A2. After preincubation with these agents, the same membranes were assayed for high-affinity opioid receptor binding [3H-labeled D-alanine-4-N-methylphenylalanine-5-glycine-ol-enkephalin (mu), 3H-labeled 2-D-serine-5-L-leucine-6-L-threonine enkephalin (delta), and [3H]ethylketocylazocine (EKC) sites] and opioid-inhibited adenylyl cyclase. Although most agents produced persistent blockade in binding of ligands to high-affinity mu, delta, and EKC sites, no change in opioid-inhibited adenylyl cyclase was detected. In most treated membranes, both the IC50 and the maximal inhibition of adenylyl cyclase by opioid agonists were identical to values in untreated membranes. Only beta-CNA blocked opioid-inhibited adenylyl cyclase by decreasing maximal inhibition and increasing the IC50 of opioid agonists. This effect of beta-CNA was not due to nonspecific interactions with G(i), Gs, or the catalytic unit of adenylyl cyclase, as neither guanylylimidodiphosphate-inhibited, NaF-stimulated, nor forskolin-stimulated activity was altered by beta-CNA pretreatment. Phospholipase A2 decreased opioid-inhibited adenylyl cyclase only when the enzyme was incubated with brain membranes in the presence of NaCl and GTP. These results confirm that the receptors that inhibit adenylyl cyclase in brain do not correspond to the high-affinity mu, delta, or EKC sites identified in brain by traditional binding studies.

Adenylyl Cyclase Inhibitors↗