Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 24
Filter
1.
Occup Med (Lond) ; 70(4): 224-230, 2020 06 20.
Article in English | MEDLINE | ID: mdl-32377669

ABSTRACT

BACKGROUND: Few fire departments in Québec have a diversified health promotion programme. Yet, many allow firefighters to physically train during working hours. AIMS: To compare the weekly physical activity (PA) level and cardiovascular health indicators of firefighters who physically train on duty to those who do not. METHODS: Participants underwent a cardiovascular health assessment and completed an online questionnaire. RESULTS: One hundred and five full-time male firefighters participated in the study. Two groups were formed: firefighters who physically train while on duty (E, n = 64) and firefighters who do not (NoE, n = 41). Following statistical adjustments, off-duty weekly PA was not different between the two groups (E: 239 ± 224 versus NoE: 269 ± 249 min, P = 0.496); however, total weekly PA was higher (P = 0.035) in E (381 ± 288 min) than in NoE (274 ± 200 min). A difference was also observed in obesity prevalence measured with waist circumference (E: 9% versus NoE: 27%, P = 0.026) and in physical inactivity prevalence (E: 0% versus NoE: 27%, P < 0.001). After statistical adjustments, E firefighters have a significantly lower waist-to-height ratio than NoE firefighters (E: 0.51 ± 0.05 versus NoE: 0.54 ± 0.05, P = 0.017). CONCLUSIONS: Results show that firefighters who physically train while on duty have a higher total PA level on a weekly basis and have better cardiovascular health indicators. Our findings suggest that fire services should promote physical training while on duty to improve firefighters' cardiovascular health.


Subject(s)
Cardiovascular Diseases/prevention & control , Firefighters/statistics & numerical data , Health Status Indicators , Occupational Diseases/prevention & control , Physical Conditioning, Human/methods , Adult , Humans , Male , Occupational Health , Preventive Health Services , Program Evaluation , Quebec
2.
Occup Med (Lond) ; 68(6): 412-414, 2018 Aug 11.
Article in English | MEDLINE | ID: mdl-29846677

ABSTRACT

BACKGROUND: Female firefighters are in the minority in the Québec firefighter population and worldwide. To our knowledge, no study has focused on cardiovascular risk factors in female firefighters, and further research in this area is needed to evaluate and reduce the risk of on-duty sudden cardiac death. AIMS: To evaluate the prevalence of cardiovascular disease (CVD) risk factors in female firefighters in Québec. METHODS: A cross-sectional study using an online questionnaire to evaluate lifestyle and CVD risk factors and symptoms. RESULTS: Forty-one female firefighters (age: 38.2 ± 9.9 years) participated in this study, representing ~7% of all female Québec firefighters. The prevalence of obesity (body mass index ≥ 30 kg/m2), hypertension, dyslipidaemia, type 2 diabetes mellitus, smoking and physical inactivity was 12% (95% confidence interval [CI] 4-26%), 5% (95% CI 0.6-19%), 5% (95% CI 0.6-19%), 3% (95% CI 0.1-14%), 14% (95% CI 5-29%) and 62% (95% CI 5-7%), respectively. Among survey participants, 76% (59-88%) had moderate to high CVD risk according to the 2013 American College of Sports Medicine guidelines. Eighty-two per cent of participants did not meet the National Fire Protection Association's required cardiorespiratory fitness standard of 12 metabolic equivalents. CONCLUSIONS: A high proportion of female firefighters in this study were at moderate to high risk of CVD. These findings suggest that they would benefit from healthy lifestyle initiatives.


Subject(s)
Cardiovascular Diseases/diagnosis , Firefighters/statistics & numerical data , Risk Assessment/methods , Adult , Body Mass Index , Cardiovascular Diseases/epidemiology , Cross-Sectional Studies , Diabetes Mellitus, Type 2/epidemiology , Dyslipidemias/epidemiology , Female , Humans , Hypertension/epidemiology , Middle Aged , Obesity/epidemiology , Occupational Health , Quebec , Risk Assessment/statistics & numerical data , Risk Factors , Surveys and Questionnaires
3.
J Fr Ophtalmol ; 38(7): 588-94, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26025033

ABSTRACT

INTRODUCTION: Selective laser trabeculoplasty (SLT) is an effective and safe procedure to lower intraocular pressure (IOP) in the management of open-angle glaucoma. The post-laser inflammatory reaction could be positively implicated in SLT efficacy and the relevance of postoperative use of topical anti-inflammatory remains controversial. The goal of this study is to determine the effect of various anti-inflammatory treatments on intraocular pressure and on side effects following SLT. MATERIAL AND METHODS: A prospective, randomized, double-blind study with a control group was conducted. Ninety-six eyes of 67 patients with primary open-angle glaucoma who underwent SLT were enrolled in this study between March 2009 and March 2012. Eyes recruited in the study were randomized to receive either prednisolone acetate 1%, diclofenac 0.1% or a placebo. The 3 treatments were administered 4 times a day for 5 days following SLT. The intraocular pressures were measured at regular intervals during the 6-months follow-up period. Side effects were also evaluated with a questionnaire as well as with the ocular exam. RESULTS: The analysis of the relative IOP decrease over the 6-months period revealed a significant difference between the time points of follow-up (P<0.0001), but no group effect (P=0.2980). No significant difference regarding anterior chamber inflammation and discomfort was observed between the 3 groups. CONCLUSION: There was no difference in intraocular pressure reduction, intraocular inflammation or ocular discomfort post-SLT when comparing the 3 treatment modalities.


Subject(s)
Anti-Inflammatory Agents/pharmacology , Diclofenac/pharmacology , Glaucoma, Open-Angle/surgery , Intraocular Pressure/drug effects , Laser Therapy , Postoperative Complications/prevention & control , Prednisolone/analogs & derivatives , Trabeculectomy , Uveitis, Anterior/prevention & control , Aged , Anti-Inflammatory Agents/adverse effects , Anti-Inflammatory Agents/therapeutic use , Anti-Inflammatory Agents, Non-Steroidal/adverse effects , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Clonidine/analogs & derivatives , Clonidine/therapeutic use , Combined Modality Therapy , Diclofenac/adverse effects , Diclofenac/therapeutic use , Double-Blind Method , Eye Pain/drug therapy , Eye Pain/prevention & control , Female , Follow-Up Studies , Glaucoma, Open-Angle/drug therapy , Humans , Male , Middle Aged , Models, Immunological , Pain, Postoperative/drug therapy , Pain, Postoperative/prevention & control , Placebos , Postoperative Complications/drug therapy , Prednisolone/adverse effects , Prednisolone/pharmacology , Prednisolone/therapeutic use , Prospective Studies , Surveys and Questionnaires , Treatment Outcome , Uveitis, Anterior/drug therapy
4.
Appl Ergon ; 47: 229-35, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25479992

ABSTRACT

Firefighting is a hazardous task associated with a heavy workload where task duration may be limited by air cylinder capacity. Increased fitness may lead to better air ventilation efficiency and task duration at a given heavy work intensity. This study compared performance, air ventilation and skeletal muscle oxygen extraction during a maximal graded walking test (GWT), a 10 METS (metabolic equivalent) treadmill test (T10) and a simulated work circuit (SWC). Participants (n = 13) who performed the SWC in a shorter time had significantly lower air cylinder ventilation values on the T10 (r = -0.495), better peak oxygen consumption (r = -0.924) during the GWT and significantly greater skeletal muscle oxygen extraction during the SWC (HbDiff, r = 0.768). These results demonstrate that the fastest participants on the SWC had better air ventilation efficiency that could prolong interventions in difficult situations requiring air cylinder use.


Subject(s)
Firefighters , Muscle, Skeletal/metabolism , Physical Exertion/physiology , Physical Fitness/physiology , Pulmonary Ventilation/physiology , Adult , Exercise Test , Humans , Male , Metabolic Equivalent , Oxygen/metabolism , Respiratory Protective Devices , Task Performance and Analysis , Time Factors , Walking/physiology , Young Adult
5.
Water Environ Res ; 74(5): 480-7, 2002.
Article in English | MEDLINE | ID: mdl-12469952

ABSTRACT

Tremendous advances have recently been made in the development of molecular tools for analysis of microbial populations in the environment. However, an appropriate scientific basis for quantification of new molecular data must exist to effectively use these tools toward increased understanding of complex waste treatment environments and implementation of corrective actions to maintain or improve system performance. In particular, molecular tools are gaining widespread use in the study of activated-sludge microbial communities and have the potential to improve monitoring and control of wastewater treatment processes. The authors have created a Web-accessible database, the Activated Sludge Biomolecular Database, which provides a scientific basis for interpreting activated-sludge biomolecular information. The database achieves its goal by accumulating and disseminating a large body of knowledge relating the presence and quantity of specific biomolecules to process design, operating conditions, and wastewater characteristics.


Subject(s)
Bioreactors , DNA, Bacterial , Databases, Factual , Sewage/microbiology , Bacteria/genetics , Internet , Reference Values , Waste Disposal, Fluid
6.
J Microbiol Methods ; 50(3): 273-82, 2002 Aug.
Article in English | MEDLINE | ID: mdl-12031577

ABSTRACT

A protocol for production, storage, and use of Shock 1 (Shk1) bioreporter cells for toxicity monitoring in wastewater treatment facilities was developed. Shk1 is a bioluminescent toxicity bioreporter for activated sludge previously constructed by the incorporation of lux genes into an activated sludge microorganism.A number of factors affecting Shk1 growth and bioluminescence were examined including the growth medium, tetracycline concentration, storage conditions, and test media. Based on the results of these experiments, a toxicity testing protocol was developed that involved growth of cultures in nutrient broth with tetracycline, storage of cultures at 4 degrees C, cell activation by reinoculation into nutrient broth, and toxicity testing by cell injection into the test media. Effective use of this approach required standardized time intervals for cell growth, storage, activation and exposure in the test media. Bioluminescence from Shk1 cells was measured in nutrient broth and influent wastewater and activated sludge mixed liquor from a municipal wastewater treatment plant. Using the Shk1 toxicity testing protocol, Zn EC(50) values for bioluminescence in nutrient broth, influent wastewater, and activated sludge mixed liquor were approximately 42, 7, and 32 mg/l, respectively. Zn concentrations as low as 1 mg/l could be detected in influent wastewater. The detection limit in influent wastewater is below the Zn concentrations typically reported to affect the activated sludge process.


Subject(s)
Hazardous Substances/analysis , Pseudomonas fluorescens , Sewage/analysis , Toxicity Tests/methods , Culture Media , Freezing , Genes, Reporter , Hazardous Substances/toxicity , Luminescent Measurements , Models, Biological , Pseudomonas fluorescens/genetics , Pseudomonas fluorescens/growth & development , Pseudomonas fluorescens/metabolism , Refrigeration , Reproducibility of Results , Sewage/chemistry , Sewage/microbiology , Zinc/metabolism , Zinc/toxicity
7.
J Cataract Refract Surg ; 27(7): 1115-8, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11489585

ABSTRACT

We report a case of bilateral keratoconus after laser in situ keratomileusis (LASIK). Before surgery, the patient had a forme fruste keratoconus, which evolved rapidly to a severe form of keratoconus in the months following LASIK. From this case, we conclude that forme fruste keratoconus is a contraindication to LASIK.


Subject(s)
Keratoconus/etiology , Keratomileusis, Laser In Situ/adverse effects , Cornea/pathology , Cornea/surgery , Corneal Topography , Humans , Male , Middle Aged , Refractive Surgical Procedures , Visual Acuity
8.
Can J Appl Physiol ; 25(4): 250-61, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10953063

ABSTRACT

Changes in physiological variables during a 60-min continuous test at maximal lactate steady state (MLSS) were studied using highly conditioned cyclists (1 female and 9 males, aged 28.3 +/- 8.1 years). To determine power at MLSS, we tested at 8-min increments and interpolated the power corresponding to a blood lactate value of 4 mmol/L. During the subsequent 60-min exercise at MLSS, we observed a sequential increase of physiological parameters, in contrast to stable blood lactate. Heart rate drifted upward from beginning to end of exercise. This became statistically significant after 30 min. From 10-60 min of exercise, a change of +12.6 +/- 3.2 bpm was noted. Significant drift was seen after 30 min for the respiratory exchange ratio, after 40 min for the rate of perceived exertion using the Borg scale, and after 50 min for % VO(2)max/kg and minute ventilation. This slow component of VO(2)max may be the result of higher recruitment of type II fibers.


Subject(s)
Bicycling/physiology , Lactic Acid/blood , Physical Exertion/physiology , Adult , Analysis of Variance , Exercise Test , Exercise Tolerance , Female , Heart Rate/physiology , Humans , Male , Middle Aged , Oxygen Consumption/physiology
9.
Appl Environ Microbiol ; 66(3): 1167-74, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10698787

ABSTRACT

The bacterial community structure of the activated sludge from a 25 million-gal-per-day industrial wastewater treatment plant was investigated using rRNA analysis. 16S ribosomal DNA (rDNA) libraries were created from three sludge samples taken on different dates. Partial rRNA gene sequences were obtained for 46 rDNA clones, and nearly complete 16S rRNA sequences were obtained for 18 clones. Seventeen of these clones were members of the beta subdivision, and their sequences showed high homology to sequences of known bacterial species as well as published 16S rDNA sequences from other activated sludge sources. Sixteen clones belonged to the alpha subdivision, 7 of which showed similarity to Hyphomicrobium species. This cluster was chosen for further studies due to earlier work on Hyphomicrobium sp. strain M3 isolated from this treatment plant. A nearly full-length 16S rDNA sequence was obtained from Hyphomicrobium sp. strain M3. Phylogenetic analysis revealed that Hyphomicrobium sp. strain M3 was 99% similar to Hyphomicrobium denitrificans DSM 1869(T) in Hyphomicrobium cluster II. Three of the cloned sequences from the activated sludge samples also grouped with those of Hyphomicrobium cluster II, with a 96% sequence similarity to that of Hyphomicrobium sp. strain M3. The other four cloned sequences from the activated sludge sample were more closely related to those of the Hyphomicrobium cluster I organisms (95 to 97% similarity). Whole-cell fluorescence hybridization of microorganisms in the activated sludge with genus-specific Hyphomicrobium probe S-G-Hypho-1241-a-A-19 enhanced the visualization of Hyphomicrobium and revealed that Hyphomicrobium appears to be abundant both on the outside of flocs and within the floc structure. Dot blot hybridization of activated sludge samples from 1995 with probes designed for Hyphomicrobium cluster I and Hyphomicrobium cluster II indicated that Hyphomicrobium cluster II-positive 16S rRNA dominated over Hyphomicrobium cluster I-positive 16S rRNA by 3- to 12-fold. Hyphomicrobium 16S rRNA comprised approximately 5% of the 16S rRNA in the activated sludge.


Subject(s)
Hyphomicrobium/isolation & purification , Industrial Waste , RNA, Bacterial/analysis , RNA, Ribosomal, 16S/analysis , Sewage/microbiology , Waste Disposal, Fluid , Cloning, Molecular , Ecosystem , Hyphomicrobium/classification , Hyphomicrobium/genetics , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology, Nucleic Acid
10.
J Ind Microbiol Biotechnol ; 19(4): 252-62, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9439001

ABSTRACT

Degradation of polychlorinated biphenyls (PCBs) in the environment is limited by their aqueous solubility and the degradative competence of indigenous populations. Field application vectors (FAVs) have been developed in which surfactants are used to both increase the solubility of the PCBs and support the growth of surfactant-degrading strains engineered for PCB degradation. Surfactant and PCB degradation by two recombinant strains were investigated. Pseudomonas putida IPL5 utilizes both alkylethoxylate [polyoxyethylene 10 lauryl ether (POL)] and alkylphenolethoxylate [Igepal CO-720 (IGP)] surfactants as growth substrates, but only degrades the ethoxylate moiety. The resulting degradation products from the alkyl- and alkylphenolethoxylate surfactants were 2-(dodecyloxy)ethanol and nonylphenoldiethoxylates, respectively. Ralstonia eutropha B30P4 grows on alkylethoxylate surfactants without the appearance of solvent-extractable degradation products. It also degrades the 2-(dodecyloxy)ethanol produced by strain IPL5 from the alkylethoxylate surfactants. The extent of degradation of the alkylethoxylate surfactant (POL) was greater for strain IPL5 (90%) than for B30P4 (60%) as determined by the cobaltothiocyanate active substances method (CTAS). The recombinant strain B30P4::TnPCB grew on biphenyl. In contrast, the recombinant strain IPL5::TnPCB could not grow on biphenyl, and PCB degradation was inhibited in the presence of biphenyl. The most extensive surfactant and PCB degradation was achieved by the use of both recombinant strains together in the absence of biphenyl. PCB (Aroclor 1242) and surfactant (POL) concentrations were reduced from 25 ppm and 2000 ppm, respectively, to 6.5 ppm and 225 ppm, without the accumulation of surfactant degradation products. Given the inherent complexity of commercial surfactant preparations, the use of recombinant consortia to achieve extensive surfactant and PCB degradation appears to be an environmentally acceptable and effective PCB remediation option.


Subject(s)
Polychlorinated Biphenyls/metabolism , Pseudomonas putida/metabolism , Surface-Active Agents/metabolism , Alcaligenes/genetics , Alcaligenes/metabolism , Biodegradation, Environmental , Genetic Engineering , Pseudomonas putida/genetics
11.
Appl Biochem Biotechnol ; 54(1-3): 277-90, 1995.
Article in English | MEDLINE | ID: mdl-7486982

ABSTRACT

A variety of modern biotechnical approaches are available to assist in optimizing and controlling bioremediation processes. These approaches are broad-ranging, and may include genetic engineering to improve biodegradative performance, maintenance of the environment, and process monitoring and control. In addition to direct genetic engineering strategies, molecular diagnostic and monitoring technology using DNA gene probing methods and new quantitative mRNA analytical procedures allows direct analysis of degradative capacity, activity, and response under in situ conditions. Applications of these molecular approaches in process developments for trichloroethylene (TCE), polychlorinated biphenyls (PCB), and polynuclear aromatic hydrocarbons (PAH) bio-oxidation in soils, aquifer sediments, and ground-water treatment reactors have been demonstrated. Molecular genetic technologies permit not only the development of new processes for bioremediation, but also new process monitoring, control strategies, and molecular optimization paradigms that take full advantage of vast and diverse abilities of microorganisms to destroy problem chemicals.


Subject(s)
Bacteria/isolation & purification , Molecular Probe Techniques , Soil Microbiology , Bacteria/genetics
12.
J Ind Microbiol ; 13(6): 392-401, 1994 Nov.
Article in English | MEDLINE | ID: mdl-7765670

ABSTRACT

The microbial populations in PCB-contaminated electric power substation capacitor bank soil (TVA soil) and from another PCB-contaminated site (New England soil) were compared to determine their potential to degrade PCB. Known biphenyl operon genes were used as gene probes in colony hybridizations and in dot blots of DNA extracted from the soil to monitor the presence of PCB-degrading organisms in the soils. The microbial populations in the two soils differed in that the population in New England soil was enriched by the addition of 1000 p.p.m. 2-chlorobiphenyl (2-CB) whereas the population in the TVA capacitor bank soil was not affected. PCB degradative activity in the New England soil was indicated by a 50% PCB disappearance (gas chromatography), accumulation of chlorobenzoates (HPLC), and 14CO2 evolution from 14C-2CB. The PCB-degrading bacteria in the New England soil could be identified by their positive hybridization to the bph gene probes, their ability to produce the yellow meta-cleavage product from 2,3-dihydroxybiphenyl (2,3-DHB), and the degradation of specific PCB congeners by individual isolates in resting cell assays. Although the TVA capacitor bank soil lacked effective PCB-degrading populations, addition of a PCB-degrading organism and 10,000 p.p.m. biphenyl resulted in a > 50% reduction of PCB levels. Molecular characterization of soil microbial populations in laboratory scale treatments is expected to be valuable in the design of process monitoring and performance verification approaches for full scale bioremediation.


Subject(s)
Polychlorinated Biphenyls/metabolism , Pseudomonas/metabolism , Soil Microbiology , Soil Pollutants/metabolism , Biodegradation, Environmental/drug effects , Chlorobenzoates/pharmacology , DNA, Bacterial/genetics , New England , Polychlorinated Biphenyls/analysis , Pseudomonas/drug effects , Pseudomonas/genetics , Pseudomonas/growth & development , Soil Pollutants/analysis , Tennessee , Time Factors
13.
Appl Environ Microbiol ; 60(8): 2826-33, 1994 Aug.
Article in English | MEDLINE | ID: mdl-8085825

ABSTRACT

Polychlorinated biphenyl (PCB)-degradative genes, under the control of a constitutive promoter, were cloned into a broad-host-range plasmid and a transposon. These constructs were inserted into a surfactant-utilizing strain, Pseudomonas putida IPL5, to create a field application vector (FAV) in which a surfactant-degrading organism cometabolizes PCB. By utilizing a surfactant not readily available to indigenous populations and a constitutive promoter, selective growth and PCB-degradative gene expression are decoupled from biphenyl. Since PCB degradation via the biphenyl degradation pathway is nonadaptive in the absence of biphenyl, there is no selective pressure for PCB gene maintenance. The recombinant strains exhibited degradative activity against 25 of 33 PCB congeners in Aroclor 1248 in the absence of biphenyl. Whole-cell enzyme assays indicated that PCB-degradative activity of a recombinant strain carrying the PCB genes on a plasmid was approximately twice that of the same strain carrying the PCB genes on a transposon. Plasmid loss rates in the absence of antibiotic selection averaged 7.4% per cell division and were highly variable between experiments. Surfactant-amended slurries of PCB-contaminated electric power plant substation soil were inoculated with approximately 10(5) recombinant cells per ml. The populations of the added strains increased to greater than 10(9) cells per ml in 2 days, and cell growth coincided with PCB degradation. By 15 days, 50 to 60% of the indicator congener 2,3,2',5'-tetrachlorobiphenyl was degraded. The effectiveness of PCB degradation by the plasmid-containing strain depended on plasmid stability. The transposon-encoded PCB genes were much more stable, and in surfactant-amended soil slurries, PCB degradation was more consistent between experiments.


Subject(s)
Dioxygenases , Genes, Bacterial/genetics , Genetic Vectors , Polychlorinated Biphenyls/metabolism , Pseudomonas putida/genetics , Surface-Active Agents , Aroclors , Biodegradation, Environmental , DNA Transposable Elements , Genes, Bacterial/physiology , Oxidation-Reduction , Oxygenases/genetics , Oxygenases/metabolism , Pseudomonas putida/enzymology , Pseudomonas putida/metabolism , Soil Microbiology
14.
Am J Physiol ; 267(1 Pt 2): H211-6, 1994 Jul.
Article in English | MEDLINE | ID: mdl-7914064

ABSTRACT

The development of heart failure (HF) on peripheral vascular control was studied in 10 conscious dogs with measurements of cardiac output (CO) and left ventricular (LV), arterial, and right atrial pressures. At 3 wk after pacing-induced HF, CO was not decreased from 2.5 +/- 0.2 l/min, whereas LV dP/dt fell (from 2,858 +/- 71 to 1,409 +/- 69 mmHg/s) and LV end-diastolic pressure increased (from 4.8 +/- 0.4 to 27.3 +/- 1.1 mmHg) (P < 0.05). At 4-7 wk after pacing, CO was significantly decreased (to 1.6 +/- 0.1 l/min; P < 0.05), but total peripheral resistance (TPR) did not rise, despite increases in plasma norepinephrine and renin activity (P < 0.05). In the presence of ganglionic blockade, TPR was still not increased in HF. In vitro studies in isolated femoral artery segments demonstrated reduced intrinsic tone (0.028 +/- 0.007 g/mg; P < 0.05) as compared with vessels from sham-operated controls (0.124 +/- 0.023 g/mg), whereas the intracellular calcium level was not altered in HF. Thus, during the development of HF, severe contractile dysfunction precedes the fall in CO, which, in turn, precedes the rise in TPR. The delayed rise in TPR appears to involve a reduction in intrinsic peripheral vascular tone, despite neurohumoral activation.


Subject(s)
Heart Failure/physiopathology , Vascular Resistance/physiology , Animals , Cardiac Output , Dogs , Female , Femoral Artery/physiopathology , Ganglionic Blockers/pharmacology , Heart Failure/blood , Hemodynamics , Hormones/blood , In Vitro Techniques , Male , Vasomotor System/physiopathology
15.
Am J Physiol ; 266(6 Pt 1): C1544-51, 1994 Jun.
Article in English | MEDLINE | ID: mdl-8023886

ABSTRACT

Because of inherent difficulties in maintaining physiological conditions in biochemical assays, the intracellular free Ca2+ concentration ([Ca2+]i) required for activation of protein kinase C (PKC) in intact cells remains unclear. In the present study, [Ca2+]i was measured in freshly isolated vascular smooth muscle cells loaded with fura 2 while, in parallel, the distribution of the Ca(2+)-dependent alpha-PKC isoform was monitored using digital imaging microscopy. The [Ca2+]i alpha-PKC translocation threshold was determined by changing extracellular free Ca2+ concentration in steps while monitoring [Ca2+]i. In the absence of agonists, increasing [Ca2+]i caused < 25% of maximal translocation. In the presence of phenylephrine, maximum translocation occurred at [Ca2+]i > or = 198 nM. Phenylephrine augmented translocation of alpha-PKC primarily by increasing the slope of the [Ca2+]i-PKC translocation relationship. These results indicate that the [Ca2+]i threshold of alpha-PKC translocation in situ is less than that reported in most in vitro assays and are consistent with an effect of agonist-induced generation of other second messengers that cause cooperative interactions leading to translocation.


Subject(s)
Calcium/metabolism , Intracellular Membranes/metabolism , Isoenzymes/metabolism , Protein Kinase C/metabolism , Animals , Biological Transport , Differential Threshold , Ferrets , Fluorescent Antibody Technique , Fura-2 , Muscle, Smooth, Vascular/cytology , Muscle, Smooth, Vascular/metabolism , Osmolar Concentration
16.
Ann N Y Acad Sci ; 721: 407-22, 1994 May 02.
Article in English | MEDLINE | ID: mdl-8010689

ABSTRACT

Molecular diagnostic methods using DNA hybridization with specific gene probes are being developed for the monitoring of microbial populations capable of polychlorinated biphenyl (PCB) degradation in contaminated soils. Evaluation of composite samples from contaminated electrical substation soil by gas chromatography (GC) indicated that the PCBs present in the soil (approximately 200 ppm) resulted from contamination with Aroclor 1248. The PCBs have been weathered or degraded so that the lower molecular weight PCB congeners are no longer present. Microbiological and molecular site characterizations are in progress to determine the abundance of PCB degradative organisms and catabolic genes present. Cloned DNA fragments for the bphC gene (2,3-dihydroxybiphenyl dioxygenase) from the biphenyl/chlorobiphenyl degradative pathways of different organisms were used as gene probes to identify indigenous microorganisms with bphC gene sequences. In colony hybridization experiments, positive signals with the pDA251 gene probe were detected in cultures from both contaminated and uncontaminated soils. The degradative abilities of indigenous microorganisms and an added PCB-degradative bacterial strain were also monitored with [14C]4-chlorobiphenyl mineralization assays and gas chromatography of PCB residues extracted from the soils. Enrichment of the contaminated soil with biphenyl and chlorobiphenyls did not stimulate the indigenous microorganisms to degrade the soil PCB. Nevertheless, enrichment of the contaminated soil with biphenyl and chlorobiphenyl and addition of the PCB-degrading strain Alcaligenes eutrophus GG4202 did result in additional degradation of the soil PCB. The results obtained from these experiments should assist in developing and monitoring a remediation plan for these PCB-contaminated soils.


Subject(s)
Polychlorinated Biphenyls/metabolism , Soil Pollutants/metabolism , Biodegradation, Environmental , Biotechnology , Biphenyl Compounds/metabolism , DNA, Bacterial/genetics , Genes, Bacterial , Genetic Engineering , Nucleic Acid Hybridization , Pseudomonas/genetics , Pseudomonas/isolation & purification , Pseudomonas/metabolism , Soil Microbiology
17.
Appl Environ Microbiol ; 59(6): 1735-41, 1993 Jun.
Article in English | MEDLINE | ID: mdl-8328798

ABSTRACT

Field application vectors (FAVs), which are a combination of a selective substrate, a host, and a cloning vector, have been developed for the purpose of expressing foreign genes in nonsterile, competitive environments in which the gene products provide no advantage to the host. Such gene products are exemplified by the enzymes for the cometabolism of polychlorinated biphenyls (PCBs) through the biphenyl degradation pathway. Attempts to use highly competent PCB-cometabolizing strains in the environment in the absence of biphenyl have not been successful, while the addition of biphenyl is limited by its human toxicity and low water solubility. Broad-substrate-specificity PCB-degradative genes (bphABC) were cloned from a naturally occurring isolate. Pseudomonas sp. strain ENV307, into broad-host-range plasmid pRK293. The resulting PCB-degrading plasmids were transferred to the FAV host Pseudomonas paucimobilis 1IGP4, which utilizes the nontoxic, water-soluble, nonionic surfactant Igepal CO-720 as a selective growth substrate. Plasmid stability in the recombinant strains was determined in the absence of antibiotic selection. PCB-degrading activity was determined by resting cell assays. Treatment of contaminated soil (10, 100, or 1,000 ppm of Aroclor 1242) by surfactant amendment (1.0% [wt/wt]Igepal CO-720 in wet soil) and inoculation with recombinant isolates of strain 1IGP4 (approximately 4 x 10(6) cells per g of soil) resulted in degradation of many of the individual PCB congeners in the absence of biphenyl.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Genetic Vectors/genetics , Plasmids/genetics , Polychlorinated Biphenyls/metabolism , Pseudomonas/enzymology , Soil Pollutants/metabolism , Biodegradation, Environmental , Cloning, Molecular , Environmental Microbiology , Pseudomonas/genetics , Pseudomonas/growth & development
18.
Circ Res ; 72(4): 778-85, 1993 Apr.
Article in English | MEDLINE | ID: mdl-8095186

ABSTRACT

Little is known about the signaling pathways involved in alpha 2-adrenergic receptor-mediated contraction of vascular smooth muscle. In the present study, we measured intracellular Ca2+ ([Ca2+]i), myosin light chain (MLC) phosphorylation, and myofilament Ca2+ sensitivity during stimulation with the relatively selective alpha 2-agonist UK 14304. These effects were compared and contrasted with corresponding changes during depolarization by elevation of the [K+] in the bathing medium. These studies were performed using spiral strips of the rabbit saphenous vein, a tissue with a relatively high density of postsynaptic alpha 2-receptors. UK 14304 (10(-5) M) caused parallel changes in [Ca2+]i, MLC phosphorylation, and force consisting of an initial phasic, followed by a sustained steady-state response. The steady-state increase in [Ca2+]i, MLC phosphorylation, and force caused by UK 14304 in the presence of 2.5 mM extracellular Ca2+ were indistinguishable from those during 51 mM K+ depolarization. However, when extracellular Ca2+ was removed in the presence of UK 14304, [Ca2+]i and MLC phosphorylation fell to resting levels, but force remained significantly elevated above basal levels. UK 14304 caused no change in the steady-state [Ca2+]i-MLC phosphorylation relation. Thus, the [Ca2+]i sensitization of force was not caused by a sensitization of MLC phosphorylation. These results indicate that in a 2.5-mM Ca2+ bathing medium, the dominant mechanism by which alpha 2-adrenergic receptor stimulation causes an increase in vascular tone is through a relatively large increase in [Ca2+]i and MLC phosphorylation. However, in Ca(2+)-free bathing medium, a second mechanism is unmasked which appears to involve an increased Ca2+ sensitivity and is independent of myosin phosphorylation.


Subject(s)
Muscle, Smooth, Vascular/physiology , Receptors, Adrenergic, alpha/physiology , Signal Transduction/physiology , Vasoconstriction/physiology , Actin Cytoskeleton/drug effects , Adrenergic alpha-Agonists/pharmacology , Aequorin , Animals , Brimonidine Tartrate , Calcium/metabolism , Calcium/physiology , Female , Homeostasis , Male , Myosins/chemistry , Myosins/metabolism , Phosphorylation , Quinoxalines/pharmacology , Rabbits , Saphenous Vein/innervation , Saphenous Vein/metabolism
19.
Am J Physiol ; 263(3 Pt 1): C714-9, 1992 Sep.
Article in English | MEDLINE | ID: mdl-1415520

ABSTRACT

It is generally assumed that smooth muscle contraction is dependent on changes in intracellular Ca2+ concentration ([Ca2+]i); however, we have previously reported that alpha-agonist-induced contraction of aorta smooth muscle cells can occur in the absence of changes in [Ca2+]i [Collins, E. M., M. P. Walsh, and K. G. Morgan. Am. J. Physiol. 262 (Heart Circ. Physiol. 31): H754-H762, 1992]. The mechanism of this [Ca2+]i-independent contraction is controversial. We have now identified the Ca(2+)-independent protein kinase C (PKC) isoforms epsilon and zeta in ferret aorta and have used digital imaging microscopy to determine their subcellular distribution. At rest, epsilon-PKC is diffusely distributed in the cytosol, whereas zeta-PKC is concentrated in the perinuclear region; both isoforms are excluded from the nuclear space. Agonist stimulation causes a [Ca2+]i-independent translocation of epsilon-PKC to the surface membrane and of zeta-PKC to the intranuclear compartment. In comparison, ferret portal vein cells, which display a totally Ca(2+)-dependent agonist contraction, are lacking in epsilon-PKC but display perinuclear zeta-PKC, which translocates intranuclearly on activation. Thus the Ca(2+)-independent vascular contraction appears to be associated with plasmalemmal translocation of epsilon-PKC; in contrast, the intranuclear translocation of zeta-PKC may function in control of gene expression.


Subject(s)
Isoenzymes/metabolism , Muscle, Smooth, Vascular/enzymology , Protein Kinase C/metabolism , Animals , Aorta/cytology , Aorta/enzymology , Biological Transport , Calcium/physiology , Ferrets , Image Processing, Computer-Assisted , Immunoblotting , Muscle, Smooth, Vascular/cytology , Portal Vein/cytology , Portal Vein/enzymology , Tissue Distribution
SELECTION OF CITATIONS
SEARCH DETAIL
...