Biomedical subjects

Michael J Cox

Publications and source records attributed to Michael J Cox.

At least 19 recordsLinked to original sources

PubMed2026

Airway microbiome diversity, intramucosal bacteria, and spatial immunity in asthmatic adults and controls.

RATIONALE: Asthma is characterized by disruption of the thoracic airway mucosae and loss of microbial diversity. Spatial profiling of the mucosal transcriptome may systematically discover mechanisms for microbial influences on immunity. OBJECTIVES: We investigated relationships between clinical measures, microbial communities, and the host mucosal transcriptome within different strata of bronchial biopsies in subjects with and without asthma. METHODS: We performed bronchoscopy in 65 asthmatic adults and 44 healthy controls, quantifying bacterial operational taxonomic units (OTUs) in bronchial brushings by 16S ribosomal RNA (rRNA) gene amplicon sequences. Biopsy histologic features were scored blind to diagnosis. Following 16S rRNA in situ hybridization of 44 biopsies, bacterial foci were scored in epithelium, basement membrane, and stroma. Global human gene expression was quantified in epithelial and stromal compartments using digital spatial profiling. MEASUREMENTS AND MAIN RESULTS: Clinical asthma was independently predicted by basement membrane abnormalities (BaseMA), endobronchial bacterial diversity, and circulating eosinophil counts, but not by specific OTU abundances. 16S rRNA staining revealed bacteria within epithelium and mucosa of all biopsies. Intramucosal bacteria counts correlated negatively with spatially organized coexpression networks encoding antigen-specific immunity, neutrophil functions, and matrix activation, whereas BaseMA correlated positively with the adaptive immunity module. Eosinophil counts correlated with epithelial bacterial counts and senescence pathways. Clinical asthma was accompanied by upregulation of a regulatory T-cell network. CONCLUSIONS: Asthma and its related phenotypes are accompanied by complex mucosal events that extend beyond eosinophilic pathways. Components of diverse airway microbiota may modify immunity by beneficial interactions within the mucosa.

Humans↗

PubMed2006

Attributable cost of catheter-associated bloodstream infections among intensive care patients in a nonteaching hospital.

OBJECTIVE: To determine the attributable cost and length of stay of intensive care unit (ICU)-acquired, catheter-associated bloodstream infections from a hospital-based cost perspective, after adjusting for potential confounders. DESIGN: Patients admitted to the ICU between January 19, 1998, and July 31, 2000, were observed prospectively for the occurrence of catheter-associated bloodstream infections. Hospital costs were obtained from the hospital cost accounting database. SETTING: The medical and surgical ICUs at a 500-bed suburban, tertiary care hospital. PATIENTS: Patients requiring central venous catheterization while in the ICU. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We measured occurrence of catheter-associated bloodstream infection, in-hospital mortality rate, total ICU and hospital lengths of stay, and total hospital costs. Catheter-associated bloodstream infection occurred in 41 of 1,132 patients (3.6 cases per 1000 catheter days). Patients with catheter-associated bloodstream infection had significantly higher unadjusted ICU length of stay (median, 24 vs. 5 days; p < .001), hospital length of stay (median, 45 vs. 11 days; p < .001), mortality rate (21 [51%] vs. 301 [28%], p = .001), and total hospital costs (83,544 dollars vs. 23,803 dollars, p < .001). Controlling for other factors that may affect costs and lengths of stay, catheter-associated bloodstream infections resulted in an attributable cost of 11,971 dollars (95% confidence interval, 6,732 dollars-18,352 dollars), ICU length of stay of 2.41 days (95% confidence interval, 0.08-3.09 days), and hospital length of stay of 7.54 days (95% confidence interval, 3.99-11.09 days). CONCLUSIONS: Patients with catheter-associated bloodstream infection had significantly longer ICU and hospital lengths of stay, with higher unadjusted total mortality rate and hospital cost compared with uninfected patients. After adjusting for underlying severity of illness, the attributable cost of catheter-associated bloodstream infection was approximately 11,971 dollars.

Aged↗

PubMed2005

Folding efficiency is rate-limiting in dopamine D4 receptor biogenesis.

Dopamine receptors are G protein-coupled receptors that are critically involved in locomotion, reward, and cognitive processes. The D2 class of dopamine receptors (DRD2, -3, and -4) is the target for antipsychotic medication. DRD4 has been implicated in cognition, and genetic studies have found an association between a highly polymorphic repeat sequence in the human DRD4 coding region and attention deficit hyperactivity disorder. Using DRD4 as a model, we show that antipsychotics can function as potent pharmacological chaperones up-regulating receptor expression and can also rescue a non-functional DRD4 folding mutant. This chaperone-mediated up-regulation involves reduced degradation by the 26 S proteasome; likely via the stabilization of newly synthesized receptor in the endoplasmic reticulum. Dopamine itself can function as a chaperone when shuttled into the cell by means of the dopamine transporter. Furthermore, different repeat variants of DRD4 display differential sensitivity to this chaperone effect. These data suggest that folding efficiency may be rate-limiting for dopamine receptor biogenesis and that this efficiency differs between receptor variants. Consequently, the clinical profile of dopaminergic ligands, including antipsychotics, may include their ability to serve as pharmacological chaperones.

Animals↗

PubMed2005

Effect of cataract surgery incision location and intraocular lens type on ocular aberrations.

PURPOSE: To determine whether Hartmann-Shack wavefront sensing detects differences in optical performance in vivo between poly(methyl methacrylate) (PMMA) and foldable acrylic intraocular lenses (IOLs) and between clear corneal and scleral tunnel incisions and whether optical differences are manifested as differences in visual performance. SETTING: Department of Optometry, University of Bradford, West Yorkshire, United Kingdom. METHODS: This study comprised 74 subjects; 17 were phakic with no ocular pathology, 20 had implantation of a Pharmacia 722C PMMA IOL through a scleral tunnel, 21 had implantation of an Alcon AcrySof IOL through a scleral tunnel, and 16 had implantation of an AcrySof IOL through a corneal incision. Visual acuity and contrast sensitivity testing, ocular optical quality measurement using Hartmann-Shack wavefront sensing, and corneal surface measurement with a videokeratoscope were performed in all cases. RESULTS: There were significant differences between groups in the total root-mean-square (RMS) wavefront aberration over a 6.0 mm pupil (F=3.91; degrees of freedom=3,70; P<.05) mediated at the 4th-order RMS, specifically spherical and tetrafoil aberrations. The PMMA-scleral group had the least aberrations and the AcrySof-corneal group the most. For a 3.5 mm diameter pupil, the total higher-order RMS wavefront aberration was not significantly different between the groups (P>.05). There were no differences between groups in corneal shape, visual acuity, or contrast sensitivity. CONCLUSIONS: Implantation of the spherical PMMA IOL led to a slight reduction in total wavefront aberration compared to phakic eyes. AcrySof IOLs induced more aberrations, especially spherical aberration. Corneal-based incisions for IOL implantation compounded this increase. Studies of the optical performance of IOLs in vivo should use wavefront sensing as the main outcome measure rather than visual measures, which are readily confounded by multiple factors.

Acrylic Resins↗

PubMed2005

Aminobacter ciceronei sp. nov. and Aminobacter lissarensis sp. nov., isolated from various terrestrial environments.

The bacterial strains IMB-1(T) and CC495(T), which are capable of growth on methyl chloride (CH(3)Cl, chloromethane) and methyl bromide (CH(3)Br, bromomethane), were isolated from agricultural soil in California fumigated with CH(3)Br, and woodland soil in Northern Ireland, respectively. Two pesticide-/herbicide-degrading bacteria, strains ER2 and C147, were isolated from agricultural soil in Canada. Strain ER2 degrades N-methyl carbamate insecticides, and strain C147 degrades triazine herbicides widely used in agriculture. On the basis of their morphological, physiological and genotypic characteristics, these four strains are considered to represent two novel species of the genus Aminobacter, for which the names Aminobacter ciceronei sp. nov. (type strain IMB-1(T)=ATCC 202197(T)=CIP 108660(T)=CCUG 50580(T); strains ER2 and C147) and Aminobacter lissarensis sp. nov. (type strain CC495(T)=NCIMB 13798(T)=CIP 108661(T)=CCUG 50579(T)) are proposed.

Agriculture↗

PubMed2005

Use of DNA-stable isotope probing and functional gene probes to investigate the diversity of methyl chloride-utilizing bacteria in soil.

Enrichment and isolation of methyl chloride-utilizing bacteria from various terrestrial environments, including woodland and forest soils, resulted in the identification of seven methyl chloride-utilizing strains belonging to the genus Hyphomicrobium, an Aminobacter strain TW23 and strain WG1, which grouped closely with the genus Mesorhizobium. Methyl chloride enrichment cultures were dominated by Hyphomicrobium species, indicating that these bacteria were most suited to growth under the enrichment and isolation conditions used. However, the application of culture-independent techniques such as DNA-stable isotope probing and the use of a functional gene probe targeting cmuA, which encodes the methyltransferase catalysing the first step in bacterial methyl chloride metabolism, indicated a greater diversity of methyl chloride-utilizing bacteria in the terrestrial environment, compared with the diversity of soil isolates obtained via the enrichment and isolation procedure. It also revealed the presence of as yet uncultured and potentially novel methyl chloride-degrading bacteria in soil.

Bacterial Proteins↗

PubMed2004

Attenuation of oxidative stress and remodeling by cardiac inhibitor of metalloproteinase protein transfer.

BACKGROUND: Matrix metalloproteinase (MMP) and cardiac inhibitor of metalloproteinase (CIMP) are coexpressed in the heart. Although it is known that oxidative stress activates MMP and CIMP inhibits MMP, it is unclear whether CIMP administration attenuates oxidative stress and MMP-mediated cardiac dilatation. METHODS AND RESULTS: Arteriovenous fistula (AVF) was created in C57BL/J6 mice, and CIMP was administered to AVF and sham mice by protein transfer into peritoneal cavity by minipump for 4 weeks. Mice were grouped as follows: sham; sham+CIMP; AVF; and AVF+CIMP (n=6). In vivo left ventricular (LV) pressure was measured. Plasma and LV tissue levels of CIMP were measured by Western analysis. LV levels of NADPH oxidase activity, marker of oxidative stress, were increased in AVF mice and decreased in AVF mice treated with CIMP. Compared with sham, CIMP was decreased in AVF mice, and CIMP protein transfer increased plasma and LV tissue levels of CIMP in AVF mice; there was no increase in sham animals. In situ zymography demonstrated a robust increase in MMP activity in the hearts from AVF mice compared with sham, and treatment with CIMP decreased MMP activity. In AVF mice, the cardiac pressure-length relationship was similar to that observed in sham mice after administration of CIMP. Contractile responses of normal LV rings were measured in the presence and absence of CIMP. CIMP shifted the pressure-length relationship to the left, attenuated LV dilatation, and had no effect on CaCl2-mediated contraction. CONCLUSIONS: Treatment of AVF mice with CIMP significantly abrogated the contractile dysfunction and decreased the oxidative stress in volume overload-induced heart failure.

Animals↗

PubMed2004

Correcting ocular spherical aberration with soft contact lenses.

Following aberroscopy, aspheric front surface soft contact lenses (SCLs) were custom-made to correct spherical refractive error and ocular spherical aberration (SA) of 18 myopic and five hypermetropic subjects (age, 20.5 +/- 5 yr). On-eye residual aberrations, logMAR visual acuity, and contrast sensitivity were compared with the best-correcting spectacle lens, an equally powered standard SCL, and an SCL designed to be aberration free in air. Custom-made and spherical SCLs reduced SA (p < 0.001; p < 0.05) but did not change total root-mean-square (rms) wave-front aberration (WFA). Aberration-free SCLs increased SA (p < 0.05), coma (p < 0.05), and total rms WFA. Visual acuity remained unchanged with any of the SCL types compared with the spectacle lens correction. Contrast sensitivity at 6 cycles/degree improved with the custom-made SCLs (p < 0.05). Increased coma with aspheric lens designs and uncorrected astigmatism limit the small possible visual benefit from correcting ocular SA with SCLs.

Adult↗

PubMed2003

Scatter and its implications for the measurement of optical image quality in human eyes.

PURPOSE: To investigate the effect of scatter on measurements of wavefront aberrations and point-spread functions in a model eye. METHODS: The wavefront aberrations of a model eye were measured using Hartmann-Shack wavefront sensing and crossed-cylinder aberroscope techniques and compared with its measured point-spread function in the presence of scattering media of different concentrations. RESULTS: The point-spread functions became broader as the concentration increased. Forward light scatter on both the light path into the eye and the light path out of the eye contributed to this broadening of the point-spread function. Neither the crossed-cylinder aberroscope nor wavefront sensing, which, respectively, measure the ocular wavefront aberrations for light entering the eye and leaving the eye, were affected by the scatter. CONCLUSION: We predict that by minimizing the contribution of the forward light scatter from one or other of these light paths by manipulating the size of the entrance and exit pupils, it should be possible to objectively assess narrow-angle forward light scatter in the eye by measuring and removing any confounding effect from wavefront aberration.

Diagnostic Techniques, Ophthalmological↗

PubMed2003

On- and off-eye spherical aberration of soft contact lenses and consequent changes of effective lens power.

BACKGROUND: Soft contact lenses produce a significant level of spherical aberration affecting their power on-eye. A simple model assuming that a thin soft contact lens aligns to the cornea predicts that these effects are similar on-eye and off-eye. METHODS: The wavefront aberration for 17 eyes and 33 soft contact lenses on-eye was measured with a Shack-Hartmann wavefront sensor. The Zernike coefficients describing the on-eye spherical aberration of the soft contact lens were compared with off-eye ray-tracing results. Paraxial and effective lens power changes were determined. RESULTS: The model predicts the on-eye spherical aberration of soft contact lenses closely. The resulting power change for a +/- 7.00 D spherical soft contact lens is +/- 0.5 D for a 6-mm pupil diameter and +/- 0.1 D for a 3-mm pupil diameter. Power change is negligible for soft contact lenses corrected for off-eye spherical aberration. CONCLUSIONS: For thin soft contact lenses, the level of spherical aberration and the consequent power change is similar on-eye and off-eye. Soft contact lenses corrected for spherical aberration in air will be expected to be aberration-free on-eye and produce only negligibly small power changes. For soft contact lenses without aberration correction, for higher levels of ametropia and large pupils, the soft contact lens power should be determined with trial lenses with their power and p value similar to the prescribed lens. The benefit of soft contact lenses corrected for spherical aberration depends on the level of ocular spherical aberration.

Adult↗

PubMed2003

Wavefront aberration and its relationship to the accommodative stimulus-response function in myopic subjects.

BACKGROUND: Autorefractors are increasingly used in myopia research because they are convenient tools to investigate aspects of the accommodation response. The degree to which the autorefractor measures are affected by ocular aberrations has been highlighted by studies that have shown changes in aberration levels through different parts of the pupil and with accommodation. We have compared accommodative accuracy as measured with a Shin-Nippon SRW 5000 autorefractor with wavefront error as measured with a Hartmann-Shack wavefront sensor to investigate how factors such as accommodation demand, ocular aberrations, and pupil size can influence autorefractor measures. METHODS: Accommodation stimulus-response curves were determined (using negative lenses) for 30 young healthy subjects (20 myopic [-0.75 to -6.00 D] and 10 emmetropic). Accommodation levels ranged from 0 to 4 D in 1 D steps. Wavefront aberrations were also determined for the same accommodation levels using a Hartmann-Shack wavefront sensor for both the subjects' natural pupil sizes and for a 2.9-mm pupil. RESULTS: For all subjects, there was a consistent increase in negative spherical aberration with increases in accommodative stimulus. However, there was no consistent change in paraxial spherocylindrical refractive correction with accommodation stimulus. For the emmetropic subjects, accommodation error as measured with the autorefractor was statistically similar to the total spherocylindrical correction for the eye as estimated by the Hartmann-Shack wavefront sensor, but only for a 2.9-mm pupil (the pupil size utilized by the autorefractor). For the myopic subjects, accommodation error as measured with the autorefractor was statistically similar to the higher-order aberrations, but only when measured for a natural pupil size. CONCLUSIONS: The relationship between the accommodation accuracy as measured with the autorefractor and the total wavefront aberration as measured with a Hartmann-Shack wavefront sensor is largely influenced by the higher-order (fourth and above) aberration levels. For the emmetropic subjects, the errors measured by the two methods agree when adjusted to measure at similar pupil sizes. For the myopic subjects with similar pupil sizes, however, the Hartmann-Shack wavefront sensor underestimates the accommodation error at higher accommodation levels (2 to 4 D) compared with the autorefractor.

Accommodation, Ocular↗

PubMed2003

Verification of aspheric contact lens back surfaces.

PURPOSE: To suggest a tolerance level for the degree of asphericity of aspheric rigid gas-permeable contact lenses and to find a simple method for its verification. METHODS: Using existing tolerances for the vertex radius, tolerance limits for eccentricity and p values and were calculated. A keratometer-based method and a method based on sag measurements were used to measure the vertex radius and eccentricity of eight concave progressively aspheric surfaces and six concave ellipsoidal surfaces. The results were compared with a gold standard measurement made using a high-precision mechanical instrument (Form Talysurf). RESULTS: The suggested tolerance for eccentricity and p value and is +/-0.05. The keratometer method was very accurate and precise at measuring the vertex radius (mean deviation +/- SD from Talysurf results, -0.002 +/- 0.008 mm). The keratometer was more precise than and similar in accuracy to the sag method for measurement of asphericity (mean deviation of keratometer method results from Talysurf results, 0.017 +/- 0.018; mean deviation of sag method results from Talysurf results using five semichords, -0.016 +/- 0.032). CONCLUSION: Neither method was precise enough to verify the asphericity within the suggested tolerance. The keratometer can be efficiently used to verify the back vertex radius within its International Organization for Standardization tolerance and the back surface asphericity within an eccentricity/p value tolerance of +/-0.1. The method is poor for progressive aspheres with large edge blending zones. Deriving the eccentricity from sag measurements is a potential alternative if the mathematical description of the surface is known. The limiting factor of this method is the accuracy and precision of individual sag measurements.

Contact Lenses↗

PubMed2003

Outcome and attributable cost of ventilator-associated pneumonia among intensive care unit patients in a suburban medical center.

OBJECTIVE: To determine the attributable cost of ventilator-associated pneumonia from a hospital-based cost perspective, after adjusting for potential confounders. DESIGN: Patients admitted between January 19, 1998, and December 31, 1999, were followed prospectively for the occurrence of ventilator-associated pneumonia. Hospital costs were defined by using the hospital cost accounting database. SETTING: The medical and surgical intensive care units at a suburban, tertiary care hospital. PATIENTS: Patients requiring >24 hrs of mechanical ventilation. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We measured occurrence of ventilator-associated pneumonia, in-hospital mortality rate, total intensive care unit (ICU) and hospital lengths of stay (LOS), and total hospital cost per patient. Ventilator-associated pneumonia occurred in 127 of 819 patients (15.5%). Compared with uninfected, ventilated patients, patients with ventilator-associated pneumonia had a higher Acute Physiology and Chronic Health Evaluation II score on admission (p <.001) and were more likely to require multiple intubations (p <.001), hemodialysis (p <.001), tracheostomy (p <.001), central venous catheters (p <.001), and corticosteroids (p <.001). Patients with ventilator-associated pneumonia were more likely to be bacteremic during their ICU stay (36 [28%] vs. 22 [3%]; p <.001). Patients with ventilator-associated pneumonia had significantly higher unadjusted ICU LOS (26 vs. 4 days; p <.001), hospital LOS (38 vs. 13 days; p <.001), mortality rate (64 [50%] vs. 237 [34%]; p <.001), and hospital costs (70,568 dollars vs. 21,620 dollars, p <.001). Multiple linear regression, controlling for other factors that may affect costs, estimated the attributable cost of ventilator-associated pneumonia to be 11,897 dollars (95% confidence interval = 5,265 dollars-26,214 dollars; p <.001). CONCLUSIONS: Patients with ventilator-associated pneumonia had significantly longer ICU and hospital LOS, with higher crude hospital cost and mortality rate compared with uninfected patients. After we adjusted for underlying severity of illness, the attributable cost of ventilator-associated pneumonia was approximately 11,897 dollars.

Adolescent↗

PubMed2003

An educational intervention to prevent catheter-associated bloodstream infections in a nonteaching, community medical center.

OBJECTIVE: To evaluate the effectiveness of an evidence-based intervention to prevent catheter-associated bloodstream infections among intensive care unit patients at a nonteaching, community hospital. DESIGN: Nonrandomized pre/post observational trial. SETTING: Two intensive care units at Missouri Baptist Medical Center, Saint Louis, MO. PARTICIPANTS: Nurses and critical care physicians. INTERVENTION: A ten-page, self-study module on the prevention of catheter-associated bloodstream infections, lectures, and posters given between July and September 1999. MEASUREMENTS: The incidence of nosocomial catheter-associated bloodstream infection and patient demographics were measured for patients admitted between March 1998 and July 2000. MAIN RESULTS: Thirty cases of catheter-associated bloodstream infections during 6110 catheter-days were noted in the preintervention period (4.9 cases/1000 catheter-days) vs. 11 cases during the 5210 catheter-days in the postintervention period (2.1 cases/1000 catheter-days). The relative risk for catheter-associated infection in the postintervention period was 0.43 (95% confidence interval, 0.22-0.84). Among catheterized patients, Acute Physiology and Chronic Health Evaluation II score (25.2 preintervention vs. 25.1 postintervention; p =.86), hemodialysis (91 of 647 [14%] patients vs. 69 of 541 [13%]; p =.70), and the mean number of catheter days per patient (9.1 vs. 9.6 days; p =.46) did not differ between the pre- and postintervention periods. CONCLUSIONS: A focused, educational intervention among nurses and physicians in a nonteaching community hospital resulted in a significant, sustained reduction in the incidence of catheter-associated bloodstream infection.

Aged↗

PubMed2002

Remodeling of the left atrium in pacing-induced atrial cardiomyopathy.

Rapid atrial pacing produces atrial systolic and diastolic failure characterized by absent atrial booster pump function, increased atrial chamber stiffness, enhanced atrial conduit function, and atrial enlargement. However, the processes underlying these abnormalities are poorly understood. Therefore, we examined left atrial myocardium from dogs with rapid pacing-induced atrial failure (400 bpm for 6 weeks) and from control dogs. Western blotting was used to determine the levels of proteins involved in calcium homeostasis (SERCA 2A, phospholamban, Na+-Ca2+ exchanger). Matrix metalloproteinase (MMP) activity was measured using gelatin and casein zymography, and levels of tissue inhibitor of metalloproteinase-4 (TIMP-4) and the TIMP-4 complexed with MMPs were measured with Western blot analysis. There were no differences in SERCA 2A or Na+-Ca2+ exchanger protein levels, but phospholamban level was significantly decreased in atrial samples from rapidly paced dogs (51.2 +/- 7.8 vs. 77.0 +/- 10.0, p < 0.01). The activity of MMP-9 was selectively and significantly increased by approximately 50%, and the level of complexed TIMP-4 protein was significantly decreased by approximately 50% in samples from dogs with atrial failure. Thus, rapid pacing-induced atrial failure is associated with differential changes in MMP activity, an unchanged number of calcium pumps, and compensatory changes in the level of phospholamban.

Animals↗

PubMed2002

Potential vision testing--the relationship between visual acuity and Vernier acuity in the presence of simulated cataract.

To evaluate the utility of a computer controlled two-bar Vernier acuity measurement as a predictor of visual function in the presence of cataract we measured logMAR visual acuity and Vernier acuity in a group of 40 young normal observers under various levels of dioptric blur (0-3 D in dioptre steps). The Vernier thresholds were resistant to dioptric blur up to 2 D, but performance degraded with blur of 3 D for non-optimised Vernier stimulus parameters. The stimulus parameters, bar length and bar separation, were further investigated in two subjects under conditions of blur. By extending the Vernier bar length and increasing the bar separation the effect of blur could be further reduced even under the most blurred condition. The relationship between visual acuity and Vernier acuity was determined. Vernier acuity was measured in the presence of Vistech cataract simulating lenses and a prediction of visual acuity was made for three observers, two with no ocular abnormality and one with age-related maculopathy. The cataract simulating lenses affected the measured visual acuity in all three subjects, but had less effect on Vernier acuity. Predicted visual acuities were all within six letters (0.12 log units) of the visual acuity without the simulated cataract. As expected, the subject sufferng from age-related maculopathy, whilst showing similar levels of Vernier acuity to the two ocularly healthy subjects at 1.5 degrees of retinal eccentricity, showed much poorer Vernier acuity for stimuli presented at fixation.

Adult↗

PubMed2002

Generation of nitrotyrosine precedes activation of metalloproteinase in myocardium of hyperhomocysteinemic rats.

The hypothesis is that homocysteine decreases endothelial nitric oxide (NO) availability by generating nitrotyrosine. In the absence of NO, and in an attempt to reduce endocardial load by dilatation, the matrix metalloproteinase (MMP) is activated. To address this hypothesis, homocysteine (0.67 mg/ml) was administered in drinking water of Sprague-Dawley rats for 8 weeks. To elicit the reversible effects of homocysteine, homocysteine was removed from the water after 8 weeks. The plasma levels of homocysteine were 2.79 +/- 0.5 microM in control (n = 6), measured by spectrofluorometry. The levels of homocysteine increased to 22 +/- 1.3 and 17 +/- 2.8 microM following 4 (n = 6) and 8 (n = 6) weeks of homocysteine treatment, respectively. The level of homocysteine decreased to 5.8 +/- 1.0 microM (n = 6) when homocysteine was removed from the drinking water. The mean arterial pressure (MAP) of control rats was 108 +/- 10 mm Hg and increased to 128 +/- 2 and 130 +/- 3 mm Hg following 4 and 8 weeks of homocysteine treatment, respectively. When homocysteine was removed from the drinking water, the MAP was decreased to 118 +/- 3 mm Hg. Left ventricle (LV) parameters were measured by a catheter in the LV through right common carotid artery in anesthetized rats. The LV tissue was analyzed for MMP activity by zymography. Levels of nitrotyrosine and cardiospecific tissue inhibitor of metalloproteinase-4 (TIMP-4/CIMP) were measured by western blot analysis using the respective antibodies. The specific bands in zymographic gel and western blot were scanned and normalized with beta-actin. The results suggest a continuous increase in nitrotyrosine levels at 4 and 8 weeks after homocysteine administration. The removal of homocysteine did not decrease the levels of nitrotyrosine. The zymographic analysis revealed a temporal increase in MMP-2 activity from 4 to 8 weeks post homocysteine administration. However, removal of homocysteine did not decrease the MMP-2 activity. The cardiac active diastolic function, -dP/dt, was decreased at 4 weeks and stayed depressed up to 12 weeks. The end-diastolic pressure started increasing at 8 weeks; at this point the MMP-2 activity was also increased. The results suggest that in the absence of endothelial NO, and in an attempt to reduce LV load, MMP-2 is activated and CIMP is inactivated, by increasing nitrotyrosine.

Animals↗

PubMed2002

Apoptosis in the left ventricle of chronic volume overload causes endocardial endothelial dysfunction in rats.

The hypothesis is that chronic increases in left ventricular (LV) load induce oxidative stress and latent matrix metalloproteinase (MMP) is activated, allowing the heart to dilate in the absence of endothelial nitric oxide (NO) and thereby reduce filling pressure. To create volume overload, an arteriovenous (A-V) fistula was placed in male Sprague-Dawley rats. To decrease oxidative stress and apoptosis, 0.08 mg/ml nicotinamide (Nic) was administered in drinking water 2 days before surgery. The rats were divided into the following groups: 1) A-V fistula, 2) A-V fistula + Nic, 3) sham operated, 4) sham + Nic, and 5) control (unoperated); n = 6 rats/group. After 4 wk, hemodynamic parameters were measured in anesthetized rats. The heart was removed and weighed, and LV tissue homogeneates were prepared. A-V fistula caused an increase in heart weight, lung weight, and end-diastolic pressure compared with the sham group. The levels of malondialdehyde (MDA; a marker of oxidative stress) was 6.60 +/- 0.23 ng/mg protein and NO was 6.87 +/- 1.21 nmol/l in the LV of A-V fistula rats by spectrophometry. Nic treatment increased NO to 13.88 +/- 2.5 nmol/l and decreased MDA to 3.54 +/- 0.34 ng/mg protein (P = 0.005). Zymographic levels of MMP-2 were increased, as were protein levels of nitrotyrosine and collagen fragments by Western blot analysis. The inhibition of oxidative stress by Nic decreased nitrotyrosine content and MMP activity. The levels of tissue inhibitor of metalloproteinase-4 mRNA were decreased in A-V fistula rats and increased in A-V fistula rats treated with Nic by Northern blot analysis. TdT-mediated dUTP nick-end labeling-positive cells were increased in A-V fistula rats and decreased in fistula rats treated with Nic. Acetylcholine and nitroprusside responses in cardiac rings prepared from the above groups of rats suggest impaired endothelial-dependent cardiac relaxation. Treatment with Nic improves cardiac relaxation. The results suggest that an increase in the oxidative stress and generation of nitrotyrosine are, in part, responsible for the activation of metalloproteinase and decreased endocardial endothelial function in chronic LV volume overload.

Animals↗