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1.
Mol Ecol Resour ; 13(4): 674-87, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23496907

ABSTRACT

Massively parallel sequencing has enabled the creation of novel, in-depth genetic tools for nonmodel, ecologically important organisms. We present the de novo transcriptome sequencing, analysis and microarray development for a vertebrate herbivore, the woodrat (Neotoma spp.). This genus is of ecological and evolutionary interest, especially with respect to ingestion and hepatic metabolism of potentially toxic plant secondary compounds. We generated a liver transcriptome of the desert woodrat (Neotoma lepida) using the Roche 454 platform. The assembled contigs were well annotated using rodent references (99.7% annotation), and biotransformation function was reflected in the gene ontology. The transcriptome was used to develop a custom microarray (eArray, Agilent). We tested the microarray with three experiments: one across species with similar habitat (thus, dietary) niches, one across species with different habitat niches and one across populations within a species. The resulting one-colour arrays had high technical and biological quality. Probes designed from the woodrat transcriptome performed significantly better than functionally similar probes from the Norway rat (Rattus norvegicus). There were a multitude of expression differences across the woodrat treatments, many of which related to biotransformation processes and activities. The pattern and function of the differences indicate shared ecological pressures, and not merely phylogenetic distance, play an important role in shaping gene expression profiles of woodrat species and populations. The quality and functionality of the woodrat transcriptome and custom microarray suggest these tools will be valuable for expanding the scope of herbivore biology, as well as the exploration of conceptual topics in ecology.


Subject(s)
High-Throughput Nucleotide Sequencing/methods , Microarray Analysis/methods , Sigmodontinae/genetics , Transcriptome , Animals , Ecology , Herbivory/genetics , Molecular Sequence Data , Norway
2.
Minerva Med ; 96(4): 223-32, 2005 Aug.
Article in English | MEDLINE | ID: mdl-16179890

ABSTRACT

Insulin resistance (IR) has profound, negative effects on the function of arteries and arterioles throughout the body and leads to arterial hypertension and vascular occlusive diseases such as heart attacks and strokes. IR affects arteries and arterioles at both the endothelium and smooth muscle levels. One major, underlying mechanism of vascular dysfunction appears to involve the augmented generation, availability and subsequent actions of reactive oxygen species (ROS). Thus, application of superoxide dismutase (SOD), a specific scavenger of superoxide anion, is able to immediately restore normal dilator responsiveness in IR arteries. In some but not all circulations, however, other factors such as increased production of and actions by constrictor agents such as endothelin also appear to restrict normal dilator responses. The basis of ROS-mediated vascular dysfunction in IR is not completely understood, but inflammatory processes throughout the arterial wall appear to be involved. Treatments involving behavioral approaches, such as changes in diet, weight loss, and regular exercise, and pharmacological approaches, involving the use of insulin-sensitizing agents or statins, appear to offer benefits against the detrimental vascular effects of IR. Nonetheless, the most effective approach appears to involve prevention of IR via adoption of a healthy lifestyle by young people.


Subject(s)
Hypertension/etiology , Insulin Resistance/physiology , Animals , Diabetes Mellitus, Type 2/prevention & control , Endothelium, Vascular/metabolism , Endothelium, Vascular/physiopathology , Humans , Hypertension/physiopathology , Muscle, Smooth, Vascular/physiopathology , Potassium Channels/physiology , Rats , Rats, Zucker , Reactive Oxygen Species/metabolism , Superoxide Dismutase/pharmacology , Vascular Resistance/physiology , Vasodilation/physiology , Vasodilator Agents/pharmacology
3.
J Oral Rehabil ; 32(8): 620-2, 2005 Aug.
Article in English | MEDLINE | ID: mdl-16011642

ABSTRACT

Patients with cleft palates eventually require definitive fixed or removable dental prostheses after the maintenance of arch alignment and occlusal relationship during adolescence. This case report presents application of a resin composite veneered fixed partial denture utilized as a definitive prosthesis for a bilateral cleft palate patient after stable occlusion had been established orthodontically. The composite veneered long span fixed partial denture provides adequate aesthetics and function.


Subject(s)
Cleft Palate/therapy , Denture, Partial, Fixed , Jaw, Edentulous, Partially/therapy , Adult , Composite Resins , Dental Veneers , Denture Design , Female , Humans , Malocclusion/therapy
4.
Am J Physiol Heart Circ Physiol ; 281(4): H1524-31, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11557540

ABSTRACT

We assessed the effect of epoxyeicosatrienoic acids (EETs) in intact mesenteric arteries and Ca(2+)-activated K(+) (BK(Ca)) channels of isolated vascular smooth muscle cells from control and insulin-resistant (IR) rats. The response to 11,12-EET and 14,15-EET was assessed in small mesenteric arteries from control and IR rats in vitro. Mechanistic studies were performed in endothelium intact or denuded arteries and in the presence of pharmacological inhibitors. Moreover, EET-induced activation of the BK(Ca) channel was assessed in myocytes in both the cell-attached and the inside-out (I/O) patch-clamp configurations. In control arteries, both EET isomers induced relaxation. Relaxation was impaired by endothelium denudation, N(omega)-nitro-L-arginine, or iberiotoxin (IBTX), whereas it was abolished by IBTX + apamin or charybdotoxin + apamin. In contrast, the EETs did not relax IR arteries. In control myocytes, the EETs increased BK(Ca) activity in both configurations. Conversely, in the cell-attached mode, EETs had no effect on BK(Ca) channel activity in IR myocytes, whereas in the I/O configuration, BK(Ca) channel activity was enhanced. EETs induce relaxation in small mesenteric arteries from control rats through K(Ca) channels. In contrast, arteries from IR rats do not relax to the EETs. Patch-clamp studies suggest impaired relaxation is due to altered regulatory mechanisms of the BK(Ca) channel.


Subject(s)
8,11,14-Eicosatrienoic Acid/pharmacology , Insulin Resistance/physiology , Mesenteric Arteries/drug effects , Vasodilation , 8,11,14-Eicosatrienoic Acid/analogs & derivatives , Animals , In Vitro Techniques , Male , Patch-Clamp Techniques , Rats , Rats, Sprague-Dawley , Reference Values , Vasodilator Agents/pharmacology
5.
J Prosthet Dent ; 86(1): 24-32, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11458261

ABSTRACT

STATEMENT OF PROBLEM: Chronic implant screw loosening remains a problem in restorative practices. Some implant manufacturers have introduced abutment screws with treated surfaces in an effort to increase preload and reduce potential loosening. Purpose. This study evaluated the materials and surfaces of 4 commercially available abutment screws on preload generation. MATERIAL AND METHODS: Twenty of each of the following abutment screws-Gold-Tite (Gt), TorqTite (Tt), gold alloy (Ga), and titanium alloy (Ta)-were divided into 2 groups. Measurements were recorded for each abutment screw on a mounted 3.75 x 18 mm external hex implant with a titanium abutment. Rotational angle measurements were conducted on the 4 abutment screws at 20 and 32 Ncm. Removal torque values were recorded and used to indirectly generate preload values. Random implant block specimens were sectioned and qualitatively evaluated with an SEM. RESULTS: At 20 and 32 Ncm, the largest rotational angles were recorded for the Tt groups: 21.2 +/- 3.1 degrees and 38.1 +/- 8.7 degrees, respectively. The greatest preload values at 20 and 32 Ncm were calculated for the Gt groups: 596.8 +/- 101.2 N and 1015.3 +/- 191.2 N, respectively. SEM analysis of the 4 implant block specimens revealed mating thread contacts located in the middle portion of the superior surface of the abutment screw thread. The greatest number of mating thread contacts were seen in the Gt implant block specimen (14 of 20 possible thread contacts). CONCLUSION: The Gt and Tt abutment screws with enhanced surfaces that help reduce the coefficient of friction produced greater rotational angles and preload values than the conventional gold alloy and titanium alloy screws.


Subject(s)
Dental Abutments , Dental Implants , Dental Materials/chemistry , Algorithms , Alloys , Analysis of Variance , Dental Alloys/chemistry , Dental Prosthesis Design , Dental Restoration Failure , Friction , Gold Alloys/chemistry , Humans , Materials Testing , Microscopy, Electron, Scanning , Palladium/chemistry , Rotation , Statistics as Topic , Stress, Mechanical , Surface Properties , Titanium/chemistry , Torque
6.
Crit Care Med ; 29(5): 1006-11, 2001 May.
Article in English | MEDLINE | ID: mdl-11378613

ABSTRACT

OBJECTIVE: Determine the effects of hypothermia on defibrillation energy requirements and cardiac electrophysiology. DESIGN: Prospective randomized acute intervention trial. SETTING: Medical center animal laboratory. SUBJECTS: Fifteen domestic farm swine. INTERVENTIONS: Swine were randomized to a hypothermia group (n = 8) or a control group (n = 7). All animals were instrumented with a transvenous defibrillation system connected to a defibrillator that delivers a biphasic-truncated waveform. Values for defibrillation energy requirements were measured at baseline (normothermia, 38-40 degrees C) and during treatment with total body hypothermia (30 degrees C) or no temperature change (sham). Hypothermia was induced by circulating ice-water through anterior and posterior surgical thermal blankets. MEASUREMENTS AND MAIN RESULTS: Defibrillation energy requirement values at 20%, 50%, and 80% were determined by using an up/down method. In the hypothermia group, defibrillation energy requirement values at baseline did not significantly change during hypothermia (defibrillation energy requirements 50% = 14 +/- 2 J vs. 15 +/- 2 J, respectively). Similarly, the defibrillation energy requirement values in the control group did not change from baseline to sham phase (defibrillation energy requirements 50% = 12 +/- 1 J vs. 13 +/- 1 J, respectively). Hypothermia profoundly affected cardiac electrophysiology, decreasing ventricular fibrillation threshold by 72%, conduction velocity by 25% (p < .01), and tissue excitability, while it prolonged ventricular repolarization and refractoriness by 7.5% to 15%, respectively (p < .05). CONCLUSIONS: Total body cooling to 30 degrees C was highly arrhythmogenic, although this unstable electrophysiological state did not alter ventricular defibrillation energy requirements. These data suggest that hypothermia may be used to slow metabolic processes without concern over the ability to successfully defibrillate and treat hypothermia-induced arrhythmias.


Subject(s)
Electric Countershock , Energy Metabolism , Hypothermia/metabolism , Ventricular Fibrillation/therapy , Animals , Electrophysiology , Swine
7.
Am J Physiol Heart Circ Physiol ; 280(2): H522-7, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11158947

ABSTRACT

Hyperinsulinemia, a primary feature of insulin resistance, is associated with increased endothelin-1 (ET-1) activity. This study determined the vascular response to ET-1 and receptor binding characteristics in small mesenteric arteries of insulin-resistant (IR) rats. Rats were randomized to control (C) (n = 32) or IR (n = 32) groups. The response to ET-1 was assessed (in vitro) in arteries with (Endo+) and without (Endo-) endothelium. In addition, arteries (Endo+) were pretreated with the ET(B) antagonist A-192621 or the ET(A) antagonist A-127722. Finally, binding characteristics of [(125)I]ET-1 were determined. Results showed that in Endo+ arteries the maximal relaxation (E(max)) to ET-1 was similar between C and IR groups; however, the concentration at 50% of maximum relaxation (EC(50)) was decreased in IR arteries. In Endo- arteries, the E(max) to ET-1 was enhanced in both groups. Pretreatment with A-192621 enhanced the E(max) and EC(50) to ET-1 in both groups. In contrast, A-127722 inhibited the ET-1 response in all arteries in a concentration-dependent manner; however, a greater ET-1 response was seen at each concentration in IR arteries. Maximal binding of [(125)I]ET-1 was increased in IR versus C arteries although the dissociation constant values were similar. In conclusion, we found the vasoconstrictor response to ET-1 is enhanced in IR arteries due to an enhanced expression of ET receptors and underlying endothelial dysfunction.


Subject(s)
Endothelin-1/metabolism , Hyperinsulinism/metabolism , Insulin Resistance/physiology , Mesenteric Arteries/metabolism , Receptors, Endothelin/metabolism , Animals , Body Weight , Dose-Response Relationship, Drug , Endothelin Receptor Antagonists , Endothelin-1/biosynthesis , In Vitro Techniques , Iodine Radioisotopes , Male , Mesenteric Arteries/drug effects , Muscle, Smooth, Vascular/drug effects , Muscle, Smooth, Vascular/metabolism , Pyrrolidines/pharmacology , Radioligand Assay , Rats , Rats, Sprague-Dawley , Receptor, Endothelin A , Receptor, Endothelin B , Receptors, Endothelin/biosynthesis
8.
J Cardiovasc Electrophysiol ; 12(12): 1363-8, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11797993

ABSTRACT

INTRODUCTION: Lidocaine increases monophasic shock defibrillation energy requirement (DER) values but does not alter biphasic shock DER values. However, the mechanism of this drug/shock waveform interaction is unknown. It may be that lidocaine increases the proarrhythmic actions of monophasic shocks but not biphasic shocks. Thus, lidocaine may increase monophasic shock DER values by increasing myocardial vulnerability to shock-induced ventricular fibrillation. METHODS AND RESULTS: Area of myocardial vulnerability (AOV), defined by a two-dimensional grid according to shock strength (y-axis) and shock coupling interval (x-axis), was assessed for biphasic shocks (n = 11) and monophasic shocks (n = 13) in intact swine hearts. Shocks were randomly delivered during right ventricular pacing at 10 shock strengths (50 to 500 V) and five coupling intervals (160 to 240 msec). AOV was defined as the number of points within the test grid that induced ventricular fibrillation. AOV, upper limit of vulnerability (ULV), and DER values were determined at baseline and during systemic infusion of lidocaine (10 mg/kg/hour). Lidocaine increased AOV, ULV, and DER values by 35%, 23%, and 36%, respectively, for monophasic shocks. However, lidocaine did not alter AOV, ULV, or DER values for biphasic shocks. CONCLUSION: Lidocaine increases the AOV to monophasic shocks, which is directly related to changes in ULV and DER values. This implies that lidocaine increases the proarrhythmic activity of monophasic shocks but not biphasic shocks. This may explain why lidocaine increases monophasic shock DER values.


Subject(s)
Anti-Arrhythmia Agents/pharmacology , Electric Countershock/methods , Lidocaine/pharmacology , Ventricular Fibrillation/etiology , Animals , Cardiac Pacing, Artificial , Electric Countershock/adverse effects , Electric Countershock/instrumentation , Electrocardiography/drug effects , Hemodynamics/drug effects , Sodium Channel Blockers , Swine
9.
J Vasc Res ; 37(5): 426-34, 2000.
Article in English | MEDLINE | ID: mdl-11025406

ABSTRACT

Impaired endothelium-dependent relaxation attributable to nitric oxide/prostacyclin-independent factor (endothelium-dependent hyperpolarizing factor; EDHF) has been demonstrated in the small mesenteric arteries of insulin-resistant rats. The purpose of this study was to determine if modulation of the cytochrome P450 enzyme system would restore EDHF-mediated relaxation in insulin-resistant rats. Sprague-Dawley rats were randomized to control (n = 32) or insulin-resistant (n = 32) groups. Each group was further randomized to treatment (n = 48) or placebo (n = 16). Miconazole (3 days) and phenobarbital (3 and 14 days) achieved cytochrome P450 inhibition and induction, respectively. Following drug treatment, mean arterial pressure was measured and vascular function was assessed in small mesenteric arteries in vitro. Specifically, acetylcholine-induced relaxation alone and in the presence of indomethacin plus N-nitro-L-arginine (LNNA) or KCl was determined. Miconazole reduced the maximal relaxation in response to acetylcholine in control rats. Similarly, in the presence of LNNA plus indomethacin, acetylcholine-induced relaxation was impaired in the miconazole-treated control group versus the placebo group, whereas relaxation in the presence of KCl was unchanged. Miconazole did not affect relaxation in insulin-resistant arteries. In contrast, 3- and 14-day treatment with phenobarbital significantly improved acetylcholine-induced relaxation in insulin-resistant arteries. Likewise, acetylcholine-mediated relaxation in the presence of LNNA plus indomethacin was also improved after phenobarbital treatment, while relaxation in the presence of KCl was unchanged. Phenobarbital treatment did not affect the control group. Miconazole treatment increased the mean arterial pressure in control rats, while 14-day phenobarbital treatment normalized the mean arterial pressure in insulin-resistant rats. Cytochrome P450 induction results in the restoration of EDHF-mediated relaxation in small mesenteric arteries and the normalization of mean arterial pressure in insulin-resistant rats. Thus, endothelial dysfunction secondary to insulin resistance can be reversed by the induction of cytochrome P450.


Subject(s)
Cytochrome P-450 Enzyme System/metabolism , Endothelium, Vascular/physiology , Insulin Resistance , Acetylcholine/pharmacology , Animals , Blood Glucose/analysis , Blood Pressure/drug effects , Enzyme Induction , Enzyme Inhibitors/pharmacology , Indomethacin/pharmacology , Insulin/blood , Male , Mesenteric Arteries , Miconazole/pharmacology , Muscle Relaxation/drug effects , Muscle, Smooth, Vascular/physiology , Nitroarginine/pharmacology , Phenobarbital/pharmacology , Potassium Chloride/pharmacology , Rats , Rats, Sprague-Dawley
10.
Cancer ; 89(6): 1349-58, 2000 Sep 15.
Article in English | MEDLINE | ID: mdl-11002231

ABSTRACT

BACKGROUND: Blacks are less likely than whites to develop bladder cancer; although once diagnosed, blacks experience poorer survival. This study sought to examine multiple biological and behavioral factors and their influence on extent of disease. METHODS: A population-based cohort of black bladder cancer patients and a random sample of frequency-matched white bladder cancer patients, stratified by age, gender, and race were identified through cancer registry systems in metropolitan Atlanta, New Orleans, and the San Francisco/Oakland area. Patients were ages 20-79 years at bladder cancer diagnosis from 1985-1987, and had no previous cancer history. Medical records were reviewed at initial diagnosis. Of the patients selected for study, a total of 77% of patients was interviewed. Grade, stage, and other variables (including age, socioeconomic status, symptom duration, and smoking history) were recorded. Extent of disease was modeled in 497 patients with urothelial carcinoma using logistic regression. RESULTS: Extent of disease at diagnosis was significantly greater in Blacks than in Whites. Older age group, higher tumor grade, larger tumors, and presence of carcinoma in situ were related to greater extent of disease in blacks and in whites. Large disparities between blacks and whites were found for socioeconomic status and source of care. Blacks had greater symptom duration and higher grade. Black women were more likely to have invasive disease than white women; this difference was not seen among men. Blacks in unskilled occupational categories, perhaps reflecting socioeconomic factors, were at much higher risk for muscle invasion than whites. CONCLUSIONS: While specific relationships between variables were noted, an overall pattern defining black and white differences in stage did not emerge. Future studies should examine the basis upon which occupation and life style factors operate by using biochemical and molecular methods to study the genetic factors involved.


Subject(s)
Black People , Carcinoma, Transitional Cell/pathology , Urinary Bladder Neoplasms/pathology , White People , Adult , Black or African American , Aged , Carcinoma, Transitional Cell/diagnosis , Carcinoma, Transitional Cell/epidemiology , Humans , Male , Middle Aged , Multivariate Analysis , Neoplasm Invasiveness , Neoplasm Staging , Smoking , Socioeconomic Factors , Urinary Bladder Neoplasms/diagnosis , Urinary Bladder Neoplasms/epidemiology
11.
IEEE Trans Biomed Eng ; 47(9): 1271-80, 2000 Sep.
Article in English | MEDLINE | ID: mdl-11008429

ABSTRACT

The procedures used to sequence the human genome involve the electrophoretic separation of mixtures of dioxyribonucleic acid (DNA) fragments tagged with reporting groups, usually fluorescent dyes. Each fluorescent pulse which arrives from an optical detector corresponds to a nucleotide (base) in the DNA sequence, and the subsequent process of base detection is known as base calling. Generating longer and more accurate sequences in the base-calling process will reduce the high cost of DNA sequencing. This paper presents an automated base-calling algorithm, referred to as maximum-likelihood base caller (MLB), which is based on maximum likelihood equalization for digital communication channels. Based on 125 experimental datasets, MLB averaged up to 40% fewer errors than the widely used ABI base caller from the Applied Biosystems Division of PE Corporation. MLB's accuracy rivaled that of another well-known base caller, Phred, surpassing it on datasets with high background noise.


Subject(s)
Likelihood Functions , Sequence Analysis, DNA/statistics & numerical data , Algorithms , Base Sequence , Biomedical Engineering , DNA/genetics , Databases, Factual , Humans
12.
Shock ; 14(1): 68-72, 2000 Jul.
Article in English | MEDLINE | ID: mdl-10909896

ABSTRACT

The splanchnic circulation constitutes a major portion of the total capacitance vasculature and may affect venous return and subsequently cardiac output during low output states. This study assessed the effects of rapid (10 microg/kg over 5 min) and slow (10 microg/kg over 60 min) induction of endotoxin (Escherichia coli) shock on splanchnic blood volume in 8 farm swine. Blood volume was measured by using Tc99m-labeled erythrocytes and radionuclide imaging. Baseline arterial pressure (MAP), central venous pressure (CVP), and liver, splenic, mesenteric and total splanchnic volumes were stable during the 30-min baseline. Approximately 30 min after the rapid endotoxin infusion, splenic volume decreased by 45%, whereas liver volume increased by 40% and MAP decreased by 60% (P < 0.01). The reduction in splenic volume occurred within 10 min of the endotoxin infusion, whereas liver volume changes occurred after MAP reduction. The slow endotoxin infusion also reduced splenic volume by approximately 50% (P = 0.05), whereas MAP declined by 30% (P < 0.05). However, the slow endotoxin infusion lowered liver volume (P < 0.05). Mesenteric volume was unaffected by the fast or slow endotoxin infusion. Total splanchnic volume was unaffected by the fast infusion but decreased by 37% in the slow infusion group (P < 0.05). In summary, E. coli endotoxin reduces splenic blood volume and increases liver blood volume after acute hypotension ensues. Endotoxin does not increase total splanchnic blood volume and may actually decrease total splanchnic volume in the absence of circulatory collapse. This endotoxin shock model is not associated with blood volume pooling in the splanchnic capacitance circulation.


Subject(s)
Endotoxemia/physiopathology , Lipopolysaccharides/toxicity , Shock, Septic/physiopathology , Splanchnic Circulation , Vascular Capacitance , Animals , Blood Volume , Drug Administration Schedule , Endotoxemia/chemically induced , Endotoxemia/diagnostic imaging , Female , Hematocrit , Hypotension/etiology , Infusions, Intravenous/methods , Lipopolysaccharides/administration & dosage , Liver/blood supply , Male , Mesentery/blood supply , Radionuclide Imaging , Shock, Septic/chemically induced , Shock, Septic/diagnostic imaging , Spleen/blood supply , Swine
13.
J Cardiovasc Electrophysiol ; 11(6): 634-41, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10868736

ABSTRACT

INTRODUCTION: Increased spatial electrical heterogeneity has been associated with impaired defibrillation efficacy. The current study investigated the relationship between electrical heterogeneity and defibrillation efficacy by manipulating spatial electrical heterogeneity. METHODS AND RESULTS: We increased spatial electrical heterogeneity by infusing potassium chloride (2 to 4 mEq/hour) or placebo in the left anterior descending artery in 13 pentobarbital anesthetized swine. Electrophysiologic measurements at five myocardial sites and defibrillation energy requirement (DER) values were determined at baseline and during regional hyperkalemia (n = 7) or placebo (n = 6). Regional potassium infusion was titrated to a 20% reduction in action potential duration in the perfused region. Regional hyperkalemia increased biphasic DER values by 87% (P = 0.02), whereas infusion of placebo did not alter defibrillation efficacy. Regional hyperkalemia decreased myocardial repolarization and refractoriness in the perfused region by 21% (P < 0.001) and 18% (P = 0.01), respectively. However, regional hyperkalemia increased ventricular fibrillation cycle length (VFCL) by 39% (P = 0.008). Consequently, dispersions of repolarization, refractoriness, and VFCL were significantly increased by 169%, 92%, and 200%, respectively. Regional hyperkalemia also increased ventricular conduction time to the perfused region by 54% (P = 0.006), indicating conduction velocity dispersion, while not affecting local pacing threshold or local voltage gradient. CONCLUSION: Regional hyperkalemia increased DER values. Regional hyperkalemia likely impairs defibrillation by increasing myocardial electrical heterogeneity, which supports the theory that electrical heterogeneity promotes nonuniform propagation of early postshock activations, thereby inhibiting defibrillation.


Subject(s)
Electric Countershock , Hyperkalemia/complications , Ventricular Fibrillation/complications , Ventricular Fibrillation/therapy , Animals , Electricity , Electrophysiology , Heart Conduction System/physiopathology , Hyperkalemia/physiopathology , Osmolar Concentration , Potassium/blood , Refractory Period, Electrophysiological , Swine , Ventricular Fibrillation/physiopathology
14.
Anal Chem ; 72(5): 1045-52, 2000 Mar 01.
Article in English | MEDLINE | ID: mdl-10739210

ABSTRACT

This paper presents results on ultralong read DNA sequencing with relatively short separation times using capillary electrophoresis with replaceable polymer matrixes. In previous work, the effectiveness of mixed replaceable solutions of linear polyacrylamide (LPA) was demonstrated, and 1000 bases were routinely obtained in less than 1 h. Substantially longer read lengths have now been achieved by a combination of improved formulation of LPA mixtures, optimization of temperature and electric field, adjustment of the sequencing reaction, and refinement of the base-caller. The average molar masses of LPA used as DNA separation matrixes were measured by gel permeation chromatography and multiangle laser light scattering. Newly formulated matrixes comprising 0.5% (w/w) 270 kDa and 2% (w/w) 10 or 17 MDa LPA raised the optimum column temperature from 60 to 70 degrees C, increasing the selectivity for large DNA fragments, while maintaining high selectivity for small fragments as well. This improved resolution was further enhanced by reducing the electric field strength from 200 to 125 V/cm. In addition, because sequencing accuracy beyond 1000 bases was diminished by the low signal from G-terminated fragments when the standard reaction protocol for a commercial dye primer kit was used, the amount of these fragments was doubled. Augmenting the base-calling expert system with rules specific for low peak resolution also had a significant effect, contributing slightly less than half of the total increase in read length. With full optimization, this read length reached up to 1300 bases (average 1250) with 98.5% accuracy in 2 h for a single-stranded M13 template.


Subject(s)
Acrylic Resins/chemistry , Electrophoresis, Capillary/methods , Sequence Analysis, DNA/methods , Base Sequence , DNA , Molecular Sequence Data , Software , Solutions
15.
Hypertension ; 35(1 Pt 1): 108-12, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10642283

ABSTRACT

This study assessed the effect of metformin treatment on insulin, mean arterial pressure (MAP), and endothelial function in insulin-resistant (IR) rats. In addition, we assessed the direct effect of metformin in vitro. Sprague-Dawley rats were randomized to control (n=28) or IR (n=28) groups. Rats were further randomized to receive metformin (300 mg/kg) or placebo for 2 weeks. MAP and insulin were measured. Subsequently, a third-order branch of the superior mesenteric artery was isolated, and endothelial function was assessed. Specifically, dose-response experiments of acetylcholine (ACh) with or without N-nitro-L-arginine (LNNA) were performed. For in vitro experiments, mesenteric arteries were removed from untreated control and IR rats and treated with metformin (100 micromol/L) before ACh+/-LNNA. MAP and insulin levels were improved in IR-metformin compared with IR-placebo rats. Maximal relaxation (E(max)) to ACh was enhanced in IR-metformin (92+/-2%) compared with IR-placebo rats (44+/-4%) (P<0.05). Relaxation in response to ACh+LNNA was greater in IR-metformin (33+/-4%) than in IR-placebo rats (12+/-4%) but remained depressed compared with control rats (E(max)=68+/-5%). The control group was not affected by metformin. In vitro treatment of arteries with metformin in response to ACh produced results similar to those in the experiments with metformin-treated rats. Although metformin improves metabolic abnormality in IR rats, this action does not appear to mediate its effect on vascular function. Both in vivo and in vitro metformin improved ACh-induced relaxation in IR rats to control levels, apparently through nitric oxide-dependent relaxation. These data suggest that metformin improves vascular function through a direct mechanism rather than by improving metabolic abnormalities.


Subject(s)
Hypoglycemic Agents/pharmacology , Insulin Resistance/physiology , Metformin/pharmacology , Acetylcholine/pharmacology , Animals , Blood Pressure/drug effects , Diabetes Mellitus, Type 2/complications , Diabetes Mellitus, Type 2/physiopathology , Diabetic Angiopathies/etiology , Endothelium, Vascular/drug effects , Endothelium, Vascular/physiology , Humans , In Vitro Techniques , Insulin/blood , Male , Nitric Oxide/physiology , Nitroarginine/pharmacology , Rats , Rats, Sprague-Dawley , Triglycerides/blood , Vasodilation/drug effects , Vasodilation/physiology
16.
Circulation ; 100(25): 2534-40, 1999.
Article in English | MEDLINE | ID: mdl-10604892

ABSTRACT

BACKGROUND: This study determined whether dispersion of conduction velocity, refractoriness, or excitability increases biphasic shock defibrillation energy requirements (DERs). METHODS AND RESULTS: Twenty-four swine were instrumented with a mid-LAD perfusion catheter for regional infusion of lidocaine 0.75 mg. kg(-1). h(-1) (n=7), low-dose d-sotalol (0.16 mg. kg(-1). h(-1)) (n=4), high-dose d-sotalol (0.5 mg. kg(-1). h(-1)) (n=6), or saline (n=7). Effective refractory periods (ERPs) were determined at 5 myocardial sites, and regional conduction velocity was determined in LAD-perfused and -nonperfused regions. Regional lidocaine infusion increased DER values by 84% (P=0.008) and slowed conduction velocity by 23% to 35% (P<0.01) but did not affect ERP. Conversely, regional low- and high-dose d-sotalol infusion did not alter DER values or conduction velocity but increased regional ERP by 14% to 17% (P<0.001). Regional lidocaine increased conduction velocity dispersion by 100% to 200% (P=0.01) but did not change ERP dispersion, whereas d-sotalol increased ERP dispersion by 140% (P<0.001) without affecting conduction velocity dispersion. Lidocaine infusion induced ventricular fibrillation (VF) in 6 of 7 animals, whereas regional d-sotalol was not proarrhythmic. Regional infusion of lidocaine and d-sotalol prolonged VF cycle length by 23% to 41% (P<0.05) in the perfused region and increased VF cycle length dispersion by 85% to 240% (P<0.05). Both agents increased pacing threshold (excitability) in the perfused region by 93% to 116% (P<0.05). CONCLUSIONS: Regional conduction velocity slowing increased DER values, which was probably a result of spatial dispersion of conduction velocity. Increasing refractory period dispersion without changing conduction velocity did not alter DFT values. Thus, dispersion of conduction velocity may be a more likely regulator of defibrillation efficacy than dispersion of refractoriness.


Subject(s)
Electric Countershock , Heart Ventricles/physiopathology , Ventricular Fibrillation/therapy , Action Potentials/drug effects , Animals , Anti-Arrhythmia Agents/pharmacology , Anti-Arrhythmia Agents/therapeutic use , Combined Modality Therapy , Electric Conductivity , Heart Ventricles/drug effects , Ion Channels/drug effects , Ion Transport/drug effects , Lidocaine/pharmacology , Lidocaine/therapeutic use , Sotalol/pharmacology , Sotalol/therapeutic use , Swine , Ventricular Fibrillation/drug therapy , Ventricular Fibrillation/physiopathology
17.
J Vasc Res ; 36(5): 385-92, 1999.
Article in English | MEDLINE | ID: mdl-10559679

ABSTRACT

OBJECTIVE: The insulin resistance syndrome is associated with atherosclerosis and cardiovascular events; however, the underlying mechanism of vascular dysfunction is unknown. The purpose of the current study was to assess endothelium- and smooth-muscle-mediated vasodilation in isolated coronary arteries from insulin-resistant rats and to determine whether insulin resistance alters the activity of the specific endothelium-derived relaxing factors. METHODS: Male Sprague-Dawley rats were randomized to insulin resistance or control. Insulin resistance was induced by a fructose-rich diet. After 4 weeks of diet, coronary arteries were removed and vascular function was assessed in vitro using videomicroscopy. Acetylcholine (10(-9)-3 x 10(-5) M)- or sodium-nitroprusside (10(-9)-3 x 10(-4) M)-induced relaxations were determined. To evaluate the role of the specific endothelium-derived relaxing factors, several inhibitors were used, including N-nitro-L-arginine (LNNA), charybdotoxin/apamin (CTX/apamin), and indomethacin. RESULTS: Studies with nitroprusside showed that smooth-muscle-dependent relaxation did not differ between insulin resistance and control groups. In contrast, maximal relaxation (E(max)) to acetylcholine was decreased in the insulin resistance group (56 +/- 7%) versus control (93 +/- 3%). LNNA pretreatment further impaired E(max) in the IR group from 56 +/- 7 to 17 +/- 2% (p < 0.01). In control, E(max) was only slightly impaired by LNNA (93 +/- 3 to 63 +/- 6%; p < 0.05). The addition of CTX/apamin also decreased relaxation in the control group (93 +/- 3 to 47 +/- 7%; p < 0.05), whereas relaxation in insulin-resistant rats was not affected (45 +/- 5% with CTX/apamin vs. 56 +/- 7% with acetylcholine alone, NS). Pretreatment with indomethacin did not affect relaxation in either group, while pretreatment with the combination of LNNA and CTX/ apamin completely abolished relaxation in both groups. CONCLUSIONS: Endothelium-dependent relaxation is impaired in small coronary arteries from insulin-resistant rats. The mechanism of this defect is related to a decrease in an endothelium-dependent, nitric oxide/prostanoid-independent relaxing factor or endothelium-derived hyperpolarizing factor.


Subject(s)
Coronary Vessels/physiopathology , Endothelium, Vascular/physiopathology , Insulin Resistance , Muscle, Smooth, Vascular/physiopathology , Acetylcholine/pharmacology , Animals , Apamin/pharmacology , Charybdotoxin/pharmacology , Coronary Vessels/drug effects , Endothelium, Vascular/drug effects , Enzyme Inhibitors/pharmacology , In Vitro Techniques , Male , Muscle Relaxation/drug effects , Muscle, Smooth, Vascular/drug effects , Nitroarginine/pharmacology , Rats , Rats, Sprague-Dawley , Vasoconstrictor Agents/pharmacology
18.
J Cardiovasc Pharmacol ; 34(3): 461-7, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10471008

ABSTRACT

Insulin resistance (IR) is associated with endothelial dysfunction. A defect in endothelium-dependent relaxation via outward potassium conductance has been observed in mesenteric arteries from IR rats. The purpose of this study was to assess whether this defect in endothelium-dependent relaxation was due to impaired endothelium-derived hyperpolarizing factor (EDHF) and to determine which specific potassium channel(s) are involved in relaxation. This was accomplished by using specific potassium channel inhibitors in the presence of nitric oxide synthase and cyclooxygenase inhibition. In addition, we sought to assess the function of smooth muscle cell adenosine triphosphate (ATP)-dependent potassium (K(ATP)) channels. Sprague-Dawley rats were randomized to control or IR. To determine EDHF-mediated relaxation, acetylcholine (ACh)-induced (10(-9)-10(-5) M) relaxation was measured (in vitro) in mesenteric arteries in the presence of indomethacin (10(-5) M) and N-nitro-L-arginine (L-NNA) (10(-4) M). Subsequently the combination of charybdotoxin (CTX) (0.1 microM) and apamin (0.5 microM) or glibenclamide (Glib) (10 microM) was added to the bath to inhibit KCa or K(ATP), respectively. In separate experiments, relaxation to pinacidil (10(-13)-10(-5) M), a K(ATP) activator, was assessed in vessels with intact endothelium, endothelium denuded, or with L-NNA. Maximal relaxation to ACh in the presence of L-NNA and indomethacin was 68+/-6% for control and 12+/-3% for IR (p<0.01). The addition of CTX + apamin almost abolished EDHF-mediated relaxation in control (Emax, 8+/-5% vs. 68+/-6%; p<0.01), whereas Glib had little affect. Neither CTX + apamin nor Glib had any affect on IR. Additionally, IR arteries were less sensitive to pinacidil than were controls (EC50, 1.5+/-0.9 microM vs. 5x10(-4)+/-3x10(-4) microM, respectively; p<0.01). Endothelial removal or L-NNA pretreatment of control arteries decreased the response to pinacidil similar to IR, whereas IR vessels were unaffected. EDHF-mediated relaxation is impaired in IR arteries. In addition, the K(Ca) channel appears to be imperative for activity of EDHF in rat small mesenteric arteries. Moreover, activation of K(ATP) channels by pinacidil is impaired in IR, and this appears to be a result of endothelial dysfunction.


Subject(s)
Biological Factors/metabolism , Endothelium, Vascular/physiopathology , Insulin Resistance , Membrane Proteins/metabolism , Potassium Channels/metabolism , Vasodilation , Animals , Diet , Endothelium, Vascular/metabolism , Fructose/pharmacology , Male , Mesenteric Arteries , Muscle, Smooth, Vascular/metabolism , Muscle, Smooth, Vascular/physiopathology , Rats , Rats, Sprague-Dawley
19.
J Cardiovasc Pharmacol ; 33(5): 698-702, 1999 May.
Article in English | MEDLINE | ID: mdl-10226855

ABSTRACT

Insulin resistance, without frank diabetes, is associated with sudden cardiac death. We postulated that a potential mechanism for this association is autonomic dysfunction. Male Sprague-Dawley rats were randomized into one of two groups: (a) insulin resistant (IR; n = 15), or (b) control (n = 11). Animals were made insulin resistant with a fructose-rich diet, whereas control animals received standard rat chow. Four weeks after randomization, arterial pressure and baroreceptor reflex were assessed. Baroreflex sensitivity was defined as the heart-rate response to acute blood pressure changes caused by nitroprusside (0.5-18 micrograms) or phenylephrine (0.2-3 micrograms). To determine the role of vagal stimulation specifically, each animal was randomized to receive atropine sulfate (1 mg/kg) or vehicle (normal saline) before administration of phenylephrine. Mean arterial pressure and fasting insulin concentrations were increased in the insulin-resistant group, whereas there were no differences in body weight, fasting glucose concentrations, or resting heart rate. Phenylephrine increased arterial blood pressure to a maximum of 54 +/- 2 mm Hg for control and 45 +/- 6 mm Hg for IR, p = 0.7. The maximal heart-rate change response to the increased blood pressure was markedly blunted in IR as compared with control (-88 +/- 12 beats/min for IR vs. -238 +/- 18 beats/min for control; p < 0.001). Thus the baroreflex sensitivity (BRS) was threefold less in IR versus the control group (-1.8 +/- 0.2 vs. -4.6 +/- 0.7 beats/min/mm Hg; p = 0.001). Pretreatment with atropine sulfate decreased the BRS in both groups, eliminating the difference between groups (-0.96 +/- 0.5 beats/min/mm Hg for control and -0.56 +/- 0.3 beats/min/mm Hg for IR; p = 0.2). Thus atropine sulfate caused the phenylephrine-induced heart rate and arterial blood pressure response to be equal between groups. On the other hand, BRS to nitroprusside-induced blood pressure changes were similar between groups. Insulin resistance, without the confounding factors of obesity, diabetes, and significant hypertension, is associated with a large reduction in vagal activity, which occurs via attenuation in reflex activity. In contrast, the insulin-resistant syndrome does not affect baroreflex sensitivity via sympathetic reflex.


Subject(s)
Baroreflex , Hyperinsulinism/physiopathology , Insulin Resistance , Vagus Nerve/physiopathology , Animals , Blood Pressure/drug effects , Heart Rate/drug effects , Male , Nitroprusside/pharmacology , Phenylephrine/pharmacology , Random Allocation , Rats , Rats, Sprague-Dawley , Vasoconstrictor Agents/pharmacology , Vasodilator Agents/pharmacology
20.
J Prosthet Dent ; 81(3): 356-64, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10050125

ABSTRACT

STATEMENT OF PROBLEM: Metal ceramic systems are advocated when both esthetics and strength are required. A major drawback to many porcelains is their wear and destruction of opposing natural dentition. PURPOSE: This study evaluated the wear of enamel opposing 1 traditional and 2 recently introduced low-fusing feldspathic dental porcelains. MATERIAL AND METHODS: Six blocks of Ceramco II, Finesse, and Omega 900 feldspathic porcelain materials were fabricated and fired according to manufacturer recommendations. Porcelain blocks were polished through 0.25 microm diamond polishing paste. Thirty-six enamel specimens were obtained and milled to a 2 mm (+0.5 mm) diameter. Enamel specimens were attached to an offset cam motor operating at 60 Hz. Enamel specimens were in constant contact with the stationary porcelain blocks under a load of 600 g and traversed a distance of 8 mm. Diameter and height of enamel specimens were measured at times of 0, 6, 12, 24, and 48 hours and subsequent determination of enamel volume loss recorded. Profilometric surface roughness of the ceramic blocks was also measured for the corresponding times. RESULTS: Statistical analysis revealed that both Finesse and Omega 900 feldspathic porcelains caused significantly less enamel volume loss when compared with Ceramco II porcelain at all time intervals. Surface roughness revealed no consistent significant differences among porcelains. CONCLUSION: Both Finesse and Omega 900 porcelains were not as destructive to human tooth structure as Ceramco II porcelain. These results suggest an advantage of the new, lower-fusing porcelains in conditions where natural dentition wear is a concern.


Subject(s)
Dental Enamel/pathology , Dental Porcelain , Tooth Attrition/etiology , Aluminum Silicates/chemistry , Analysis of Variance , Dental Enamel/ultrastructure , Dental Polishing , Dental Porcelain/chemistry , Humans , Materials Testing , Microscopy, Electron, Scanning , Potassium Compounds/chemistry , Stress, Mechanical , Surface Properties , Time Factors , Tooth Attrition/pathology
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