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Of the patients with end-stage cardiomyopathy on a heart transplant list, 95% do not receive a donor heart. Due to this severe shortage of donor organs, an artificial replacement heart has been pursued for several decades. To date, 10 patients have received an Abiocor artificial replacement heart (Abiomed, Inc, Danvers, Mass), the latest artificial heart device available.Anesthetic management often is complicated by the preoperative multisystem organ failure arising from the patients advanced cardiomyopathy. Hemodynamic stabilization before cardiopulmonary bypass is extremely challenging, and the postbyposs period creates a unique situation in which the only control the anesthesia provider has on the hemodynamics is management of the systemic vascular resistance. This article describes the anesthetic management of the recipient of the Abiocor artificial heart with the longest survival time. [ABSTRACT FROM AUTHOR]

Background: Patients undergoing facial rejuvenation surgery are at unique risk of perioperative complications from the anesthetic utilized during the procedure. The ideal anesthetic agent is one that is safe to use in the outpatient population, has analgesic, sedative, and anesthetic properties, yet does not cause respiratory depression or hemodynamic irregularities., Objectives: A retrospective analysis of a large outpatient facelift cohort was performed to determine if dexmedetomidine, an α 2-adrenergic receptor agonist, meets the criteria of an ideal adjunct for propofol in a total intravenous anesthesia protocol., Methods: The charts of 791 patients who underwent rhytidectomy with total intravenous anesthesia were reviewed and data of patients' operative risk factors, perioperative management including medications administered, perioperative vital signs, and postoperative adverse events were recorded. Statistical univariate analyses were performed on the data., Results: Dexmedetomidine resulted in a significant reduction and maintenance of blood pressure from onset of anesthesia until discharge from the postanesthetic recovery unit. The utilization of opioids and anxiolytics was significantly less than previously reported for other anesthetic types. The postoperative nausea/vomiting rate was 0.8% (6 patients). There were no postoperative admissions for inpatient management. Forty-three (5.3%) patients required a conversion to general endotracheal anesthesia and statistically significant risk factors include increased BMI, American Society of Anesthesiologists Class II or higher, preoperative hypertension, and multiple procedures performed., Conclusions: This study demonstrated the safety and efficacy of dexmedetomidine in a large cohort of outpatients undergoing facelift. Dexmedetomidine meets the requirements for an ideal adjunct anesthetic within a total intravenous anesthesia protocol., (© The Author(s) 2022. Published by Oxford University Press on behalf of The Aesthetic Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Serum macrophage inhibitory cytokine-1 (MIC-1/GDF15) concentration has been associated with colonic adenomas and carcinoma., Aims: To determine whether circulating MIC-1/GDF15 serum concentrations are higher in the presence of adenomas and whether the level decreases after excision., Methods: Patients were recruited prospectively from a single centre and stratified into five groups: no polyps (NP); hyperplastic polyps (HP); sessile serrated ademona (SSA); adenomas (AP); and colorectal carcinoma (CRC). Blood samples were collected immediately before and 4 weeks after colonoscopy. MIC-1/GDF15 serum levels were quantified using ELISA., Results: Participants (n=301) were stratified as: NP; n=116 (52%), HP; n=37 (12%), SSA; n=19 (7%), AP; n=68 (23%); and CRC; n=3 (1%). Patients were excluded from the study due to nondiagnostic pathology (n=9, 3%) and exclusion criteria (n=20, 6%). In the 272 remaining subjects (M=149; F=123), age (P=.005), history of colonic polyps (P=.003) and family history of colonic polyps (P=.002) were associated with presence of adenomas. Baseline median MIC-1/GDF15 serum levels increased significantly from NP 609 (460-797) pg/mL, HP 582 (466-852) pg/mL, SSA 561 (446-837) pg/mL to AP 723 (602-1122) pg/mL and CRC 1107 (897-1107) pg/mL; (P<.001). In the pre- and postpolypectomy paired adenoma samples median MIC-1/GDF15 reduced significantly from 722 (603-1164) pg/mL to 685 (561-944) pg/mL (P=.002). A ROC analysis for serum MIC-1/GDF15 to identify adenomatous polyps indicated an area under the curve of 0.71., Conclusions: Our data suggest that serum MIC-1/GDF15 has the diagnostic characteristics to increase the detection of colonic neoplasia and improve screening., (© 2017 John Wiley & Sons Ltd.)

Objective: To identify sociodemographic, knowledge and attitudinal correlates to antibiotic sharing among a community-based sample of adults (age 18 and older) in a low-income setting of the Philippines and to explore community-level data on informal antibiotic distribution in roadside stands (i.e., sari-sari stands)., Methods: Participants (n = 307) completed self-administered surveys. Correlates to antibiotic sharing were assessed using logistic regression with Firth's bias-adjusted estimates. Study staff also visited 106 roadside stands and collected data on availability and characteristics of antibiotics in the stands., Results: 78% had shared antibiotics in their lifetime, most often with family members. In multivariable analysis, agreement with the belief that it is safe to prematurely stop an antibiotic course (OR: 2.8, CI: 1.3-5.8) and concerns about antibiotic side effects (OR: 2.1, CI: 1.1-4.4) were significantly associated with increased odds of reported antibiotic sharing. Antibiotic sharing was not associated with sociodemographic characteristics or antibiotic knowledge. Antibiotics were widely available in 60% of sampled sari-sari stands, in which 59% of antibiotics were missing expiration dates. Amoxicillin and cephalexin were the most commonly available antibiotics for sale at the stands (60% and 21%, respectively)., Conclusions: Antibiotic sharing was common and was associated with misconceptions about proper antibiotic use. Antibiotics were widely available in sari-sari stands, and usually without expiration information. This study suggests that multipronged and locally tailored approaches to curbing informal antibiotic access are needed in the Philippines and similar Southeast-Asian countries., (© 2017 John Wiley & Sons Ltd.)

Natural transformation is the main means of horizontal genetic exchange in the obligate human pathogen Neisseria gonorrhoeae and drives the spread of antibiotic resistance and virulence determinants. Transformation can be divided into four steps: (1) DNA binding, (2) DNA uptake, (3) DNA processing and (4) DNA recombination into the chromosome. The DNA processing enzyme DprA has been shown to shuttle incoming ssDNA to the recombination enzyme RecA during transformation in Bacillus subtilis and Streptococcus pneumoniae. Here, we investigate the role of DprA during transformation in N. gonorrhoeae. Inactivation of dprA completely abrogated transformation of gyrB1-encoding DNA, which confers nalidixic acid resistance. The presence of the DNA uptake sequence enhances DNA uptake and transformation by binding to the minor pilus protein ComP. Loss of transformation in the dprA null mutants was independent of the DNA uptake sequence. DprA mutants exhibited increased RecA-dependent pilin variation suggesting that DprA affects pilin variation. Unlike the exquisite UV sensitivity of a recA mutant, inactivation of dprA did not affect survival following UV irradiation. These results demonstrate that DprA has a conserved function during transformation, and reveal additional effects of DprA in N. gonorrhoeae during pilin variation.

In the years surrounding World War II, solar house heating was seen by many American architects, journal editors, and policymakers as a necessary component of the expansion into suburbia. As the technological and financial aspects of home ownership came to take on broad social implications, design strategies of architectural modernism--including the expansive use of glass, the open plan and façade, and the flexible roof line--were seen as a means to construct suburbs that were responsive to anticipated concerns over materials allocations, over energy-resource scarcity, and over the economic challenges to postwar growth. As this article demonstrates, experiments in passive solar house design were a prominent means for envisioning the suburbs as an opportunity for new kinds of building and new ways of living. The article documents these developments and places them in the context of related efforts to think about the future.

An ecological model for transmission of Salmonella enterica in swine production ecosystems was developed, identifying host species, environmental reservoirs, and temporal, spatial, and functional (i.e., stage of production) dimensions. It was hypothesized that transmission was most likely within spatial and functional compartments, between hosts of the same species and abiotic compartments of the same type. Eighteen swine production systems in Illinois, USA, were sampled in four collection cycles (1998, 1999, 2000, 2003). There were 11,873 samples collected, including feces from swine and other mammals and birds, and samples from insects, pen floors, boots, feed, and water. The 460 Salmonella isolates obtained were genotyped using repetitive sequence PCR with three primers-REP, BOX, and ERIC. All isolates from 2000 and 2003 were serotyped, as well as a subsample from 1998 and 1998. Genetic relatedness was estimated from the similarity of fragmentation patterns after gel electrophoresis of PCR products. Cluster analysis identified genetically related isolates. Linking of isolates in tight clusters (similarity >or=85%) was viewed as evidence for transmission. Five farms had a sufficient number of tight clusters for hypothesis testing. The factors most differentiating isolates genetically were farm of origin and time of sampling. Isolates were also differentiated genetically by site, building, room, and pen. There was no consistent association of genotype with stage of production or host/environment reservoir. Serotype analysis confirmed that Salmonella lineages were differentiated by visit and site. Thus, Salmonella transmission was primarily over short distances, i.e., within the same pen or room, with some transmission between rooms and buildings on the same site, but with limited transmission between sites. Transmission was observed across a variety of ecological niches represented by different host species and environmental reservoirs. Genetic differences over time reflected multiple introductions into the ecosystem of different Salmonella genotypes, as well as evolutionary changes within lineages. Intervention strategies to reduce Salmonella prevalence within swine production ecosystems would be best targeted at maintaining spatial barriers to transmission, whereas intervention targeted at specific biological hosts or environmental reservoirs is less likely to be effective.

Salmonellosis in humans is a costly disease traditionally assumed to be associated with exposure to contaminated food. We have developed a farm-to-fork model that allows estimation of the human health costs and risks associated with Salmonella in pork. This analysis focuses on the stages of the pork production chain up to the point of producing a chilled pork carcass. The model predicts the number of human cases of salmonellosis associated with pork (mean, 99,430; 90% confidence interval, 20,970 to 245,560) and the corresponding social costs (mean, $81.53 million; 90% confidence interval, $18.75 million to $197.44 million). Sensitivity and scenario analyses suggest that changes in Salmonella status during processing are more important for human health risk and have a higher benefit:cost ratio when compared with on-farm strategies for Salmonella control. Specifically, benefit:cost ratios are less than 1 (indicating they are not likely to be profitable from a social economic perspective) for the on-farm strategies of vaccination and meal feeding, whereas rinsing carcasses at various temperatures with and without sanitizer all have benefit:cost ratios greater than 1 (indicating they are profitable from a social economic perspective). This type of modeling is useful for evaluation of the relative cost effectiveness of interventions at different points in the food chain when allocating limited food safety dollars and is best used for examining trends and alternative strategies rather than for providing definitive dollar value estimates of risk. The dollar value estimates must be considered in the context of the wide confidence intervals.

Pulsed field gel electrophoresis (PFGE) using restriction enzymes AvrII, SpeI, and XbaI, and repetitive sequence polymerase chain reaction (Rep-PCR) using BOX, ERIC, and REP primers, were compared with respect to their ability to detect genetic differences among 68 Salmonella isolates from nine Illinois swine farms. Both genotyping methods had high reproducibility of fragment numbers (reliability>0.9) and sizes (reliability>0.85), and sizes [Formula: see text], and produced approximately the same number of DNA fragments, but Rep-PCR fragment profiles had considerably greater variation. Genetic distances between isolates were calculated from fragment size matching. There was good agreement between the genetic distance matrices for the composite (3-enzyme and 3-primer) methods (Mantel's r=0.83). PFGE detected slightly greater variation in genetic distances among isolates, but failed to differentiate seven pairs of isolates, three of which were sampled at least 1 month apart and therefore unlikely to be truly identical genetically. In contrast, Rep-PCR identified no isolates as genetically identical. In cluster analyses based on genetic distances, there were moderate differences between PFGE and Rep-PCR (about 2/3 agreement in tight cluster membership). Both PFGE and Rep-PCR were able to differentiate isolates of the same serotype. However, some serotypes (Agona, Anatum, Derby, Infantis, Worthington) were distributed across clusters. There was less agreement between individual primer/enzyme and composite results for Rep-PCR than for PFGE. This greater independence of results for individual primers for Rep-PCR accounted in part for the greater discriminative ability of the composite method. Both composite methods indicated that most Salmonella transmission occurred within a farm and that there was no preference for transmission between specific ecological compartments. Given the equally high reliability of both genotyping methods, the greater discriminative ability of Rep-PCR recommends it as the preferred method for precise detection of transmission links.

Antimicrobial resistance is an issue of increasing global concern. Several investigators have suggested that antibiotic use in food-producing animals is a major contributor to the increasing incidence of antimicrobial-resistant organisms causing illness in humans (F. J. Angulo, K. R. Johnson, R. V. Tauxe, and M. L. Cohen, Microb. Drug Res. 6:77-83, 2000; P. D. Fey, T. J. Safranek, M. E. Rupp, E. F. Dunne, R. Efrain, P. C. Iwen, P. A. Bradford, F. J. Angulo, and S. H. Hinrichs, N. Engl. J. Med. 342:1242-1249, 2000; S. A. McEwen and P. J. Fedorka-Cray, Commun. Infect. Dis. 34(Suppl. 3):S93-S106, 2002; D. L. Smith, A. D. Harris, J. A. Johnson, E. K. Silbergeld, and J. G. Morris, Jr., Proc. Natl. Acad. Sci. USA 99:6434-6439, 2002; D. G. White, S. Zhao, R. Sudler, S. Ayers, S. Friedman, S. Chen, P. F. McDermott, D. D. Wagner, and J. Meng, N. Engl. J. Med. 345:1147-1154, 2001; W. Witte, Science 279:996, 1998). In this paper, we discuss this and other assumptions relevant to a quantitative risk assessment model for salmonellosis in humans. We also discuss other important aspects of modeling food safety and food-associated antimicrobial resistance risk to humans. We suggest that the role of food-producing animals in the origin and transmission of antimicrobial resistance and "foodborne" pathogens has been overestimated and overemphasized in the scientific literature; consequently, nonfoodborne transmission, including pet-associated human cases, has been underemphasized. Much evidence exists for the potential contribution to infectious disease that may be of human or pet origin (that may contact humans through food but not be of a food origin). Risk analyses that do not acknowledge the potential for these sources of cross-contamination will understate the contribution that origin has in the realm of foodborne and food-associated diseases (e.g., Salmonella) and the resulting uncertainty levels in the food system, thus leading to biased inferences. We emphasize the importance of evaluating both the foodborne and nonfoodborne transmission risk for salmonellosis and outline the basics of an analytical modeling approach in food safety with examples to illustrate strengths and limitations in the modeling. Examples illustrate, on a simplistic level, how varying assumptions and other inputs can influence the output of food-associated quantitative risk models.

The objective of this 2-year field survey was to sample multiple ecological compartments within swine production systems to identify potential sources of Salmonella infection for swine. Twelve single-site production systems within Illinois were identified by slaughter sampling to have detectable Salmonella in swine and therefore selected for study. There were four visits to each farm during a 5-month period. Fecal samples were obtained from swine and other wild and domestic mammals. Arthropods and environmental samples of feed, water, pen floors, boots, and bird feces were also collected. All 8,066 samples obtained were cultured to detect Salmonella. Salmonella was detected on 11 of the 12 farms. There were 206 positive cultures, including samples from swine (83), pen floors (54), boots (32), flies (16), mice (9), cats (3), and birds (3). Swine shedding Salmonella in feces were detected on 9 of the 12 farms. The more Salmonella-abundant ecological compartments were cats (12% of samples positive), boots (11%), bird feces (8%), flies (6%), and mice (5%); 2.1% of 4,024 swine samples were positive. All 221 feed samples were negative for Salmonella. There was a correlation between a farm having a high prevalence of shedding Salmonella in pigs and a high abundance on pen floors, flies, and boots. The most common serotypes detected were Derby, Agona, Worthington, and Uganda, which were distributed throughout the ecosystem, suggesting widespread transmission across ecological compartments. The ubiquitous distribution of Salmonella suggests that an effective control strategy must target multiple compartments of the swine production ecosystem.

Pulsed field gel electrophoresis (PFGE) using 3 enzymes (Spe I, Xba I, Avr II) and repetitive sequence polymerase chain reaction (REP-PCR) with 3 primers (BOX, ERIC, REP) were compared with respect to their validity as a method for identifying transmission of Salmonella on swine farms. Sixty-eight isolates of Salmonella were obtained from feces of swine, cats, mice, and birds, insect body parts, water and floor samples, and boot scrapings collected on 9 swine farms in Illinois USA. Genetic distances between isolates were calculated using the Dice matching coefficient. Cluster analysis of distance matrices was conducted using the UPG-MA algorithm. There was no significant difference between PFGE and REP-PCR in the genetic diversity detected; however, REP-PCR differentiated between 14 pairs of isolates which PFGE identified as identical. There were no significant differences between PFGE and REP-PCR in identifying all or most close genetic links as isolates from the same farm, the same building, and from the same sampling visit, suggesting ecological validity for both methods. Thus, REP-PCR should be considered as an acceptable and perhaps preferable alternative to PFGE as a genotyping method for studies of Salmonella transmission.

Background: Vasoconstriction in response to endothelin-1 has been shown to be primarily related to its effects on the endothelin-A receptor. Experimental hypercholesterolemia is associated with an increase in coronary vasoconstrictor response to endothelin-1 in vivo, although the relative contributions of subtypes of endothelin receptor in this model remain unknown., Objective: To test the hypothesis that there is an increase in coronary vasoconstriction in response to stimulation of endothelin-B receptor in hypercholesterolemia, which might be related to attenuation of activity of endothelin-derived relaxing factor., Methods: We infused 5 ng/kg/min sarafotoxin, a specific endothelin-B receptor agonist, or 50 micrograms/kg/min NG-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of nitric oxide synthase, into the left anterior descending coronary arteries of pigs before and after feeding them a cholesterol-rich diet for 10 weeks., Results: There was a significant increase in serum level of cholesterol. After 10 weeks, infusion of sarafotoxin resulted in an accentuated decrease in coronary blood flow (CBF) compared with baseline (decreases by 60 +/- 7 versus 34 +/- 6%, P < 0.05). There was no significant difference between the effects on diameter of coronary arteries for the two time periods. The effect of L-NMMA on CBF was attenuated after 10 weeks (by 5 +/- 10.1 versus 45.6 +/- 4.7%, P < 0.05). Endothelin-receptor status of epicardial coronary arteries remained unchanged. Sarafotoxin and L-NMMA were co-infused at the above-mentioned doses into normolipidemic animals; the decrease in CBF in response to this co-infusion was comparable to the decrease observed with sarafotoxin alone in hypercholesterolemic animals (decreases of 67 +/- 5 versus 60 +/- 7, NS)., Conclusions: The present results demonstrate that selective stimulation of the endothelin-B receptor increases coronary vasoconstriction in experimental hypercholesterolemia, primarily at the level of the microvasculature. These findings may be related to the attenuation of activity of endothelin-derived relaxing factor in this model, and support the hypothesis that endothelin-B receptor plays a role in the regulation of coronary vascular tone in pathophysiologic states.

Introducing recombinant genes into donor hearts may offer a therapeutic intervention that could potentially attenuate the complications of heart transplantation, including rejection, infection and accelerated atherosclerosis. In the cardiovascular system, reduced bioactivity of endothelial nitric oxide is a feature of atherosclerosis and vascular injury. Nitric oxide is an arterial vasodilator that also inhibits proliferation of vascular smooth muscle cells and platelet aggregation. Experiments were designed to determine the distribution of adenoviral-mediated transfer of recombinant endothelial nitric oxide synthase gene (eNOS) and the effect of recombinant gene expression on the function of transplanted hearts. Adenoviral vectors for (a) bovine eNOS (AdeNOS) or (b) beta-galactosidase (AdLacZ; control) were infused into two groups (n = 12, per group) of explanted rat hearts. The transduced hearts were then implanted heterotopically into the abdomen of syngeneic recipient rats. After four days, the hearts were excised and examined for distribution and function of the recombinant genes. Polymerase chain reaction (PCR) verified the presence of the recombinant eNOS gene in eNOS-transduced but not in beta-galactosidase-transduced hearts; reverse transcriptase-PCR identified mRNA for eNOS in AdeNOS-transduced hearts. NOS activity (conversion of tritiated L-arginine to citrulline) was greater in homogenates of AdeNOS-compared to AdLacZ-transduced hearts. Positive immunoreactivity for eNOS was present in cardiomyocytes predominantly in eNOS-transduced hearts. Myocardial contractility and coronary blood flow, as determined using a Langendorff preparation, were not different between hearts transduced with AdeNOS or AdLacZ. These results suggest that, up to four days post transplantation, adenoviral-mediated transfer of eNOS into transplanted hearts is possible. However, expression of the recombinant protein did not result in measurable changes in myocardial contractility or coronary perfusion.

1. Experiments were designed to determine whether or not interactions of platelets with coronary arteries are affected by gender or oestrogen-status. 2. Platelets and right coronary arteries were isolated from sexually mature male, female and ovariectomized pigs. Arteries were suspended in organ chambers for the measurement of isometric force. Responses of rings, with and without endothelium, were evaluated to aggregating platelets and the platelet products 5-hydroxytryptamine (5-HT) and adenosine diphosphate (ADP). 3. Release of 5-HT, thromboxane A2 (TXA2) and prostacyclin were measured from platelets. 4. Platelets caused relaxations of rings with endothelium from all pigs. However, in rings without endothelium, consistent contractions were observed only in rings from male pigs. 5. The release of 5-HT and prostacyclin was greatest from platelets of ovariectomized pigs compared with male and female pigs. Release of TXA2 was greatest from platelets of male pigs. 6. Endothelium-dependent relaxations to ADP and contractions to 5-HT were similar among the three groups. 7. These results suggest that there may be gender-specific differences in vasomotor responses to autogenous platelets but not to the platelet-derived products 5-HT and ADP. Furthermore, there are gender differences in platelets in the release of cyclo-oxygenase metabolites of arachidonic acid and 5-HT. These products could contribute to gender differences in response to injury in the coronary circulation.

Experiments were designed to determine whether or not relaxations of coronary arterial smooth muscle to C-type natriuretic peptide (CNP) vary according to gender, and if so, to determine mechanisms for the differences. Rings of coronary arteries without endothelium from sexually mature male and female Yorkshire pigs were suspended in organ chambers for measurement of isometric force. Cumulative concentration-responses to CNP (10(-9)-10(-7) M) were obtained in the absence and presence of either K+ channel blockers (charybdotoxin, apamine, or glibenclamide, 10(-7) M) or the clearance-receptor antagonist C-ANP (10(-6) M) during contractions to prostaglandin F2alpha (2 microM). Relaxations to CNP were significantly less in arteries from male compared with female pigs and were significantly attenuated by charybdotoxin and glibenclamide in both sexes. However, apamine reduced relaxations to CNP only in arteries from female pigs. C-ANP significantly potentiated relaxations to CNP only in arteries from male pigs. In separate experiments, cyclic guanosine monophosphate (cGMP) was measured by radioimmunoassay at specified times after the addition of CNP (10(-7) M). Peak increases in cGMP were greater and occurred earlier in arteries from female than from male pigs; these differences were eliminated by the phosphodiesterase inhibitor 3-isobutyl-1-methyl-xanthine (10(-4) M). These results demonstrate three mechanisms that contribute to gender differences in CNP-mediated relaxation of coronary arterial smooth muscle: activation of low conductance Ca2+-activated K+ channels, natriuretic peptide clearance receptors, and activity/regulation of phosphodiesterases.

Takayasu's arteritis, also known as Takayasu's syndrome, is a chronic inflammatory disease, which primarily affects large vessels including the aorta and its main branches. The anesthetic management for the patient presenting with Takayasu's arteritis is complicated by the multiple organ systems affected by the disease. A 59-year-old Chinese female with the diagnosis of Takayasu's arteritis presented with severe progressive occlusion of the thoracic and abdominal aorta and bilateral renal artery stenosis, necessitating an extra-anatomic aorto-aortic bypass and bilateral aorto-renal bypass. The patient tolerated the operative procedure without any untoward events. This case report discusses the pathophysiology of Takayasu's arteritis and the anesthetic management of aortic surgery specific to this particular disease process.

In arteries, adrenomedullin (ADM) causes relaxations of rings with and without endothelium by stimulating accumulation of cyclic nucleotides resulting from activation of the ADM and calcitonin gene-related peptide (CGRP) receptors. Experiments were designed to determine the mechanism(s) of relaxation to ADM in veins. Rings of canine femoral vein with and without endothelium were suspended in organ chambers for measurement of isometric force. Rings were contracted with prostaglandin F2alpha (2 x 10(-6) M), and cumulative dose-responses to ADM (10(-11) to 10(-7) M) were obtained in the absence or presence of indomethacin (10(-5) M), indomethacin + N(G)-monomethyl-L-arginine (10(-4) M), methylene blue (10(-5) M), particulate guanylate cyclase inhibitor HS-142-1 (10(-5) M), tetraethylammonium (TEA, 10(-2) M), CGRP-receptor antagonist (CGRP 8-37, 10(-6) M), ADM-receptor antagonist (ADM 26-52, 10(-6) M), diphenhydramine (10(-6) M), 8-phenyltheophylline (3 x 10(-6) M), or superoxide dismutase (150 U/ml) plus catalase (1,200 U/ml). ADM produced concentration-dependent relaxations only in veins with endothelium. Relaxations to ADM in rings with endothelium were significantly inhibited only by methylene blue and HS-142-1. In separate experiments, incubation of rings with ADM (10(-8) M) and 3-isobutyl-1-methyl-xanthine (10(-4) M) for 3 min did not significantly affect the accumulation of cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP). These data suggest that ADM-mediated relaxation in veins is endothelium dependent and is not associated with activation of CGRP receptors or currently defined ADM receptors. Further, relaxations are not mediated by nitric oxide, indomethacin-sensitive prostanoids, TEA-sensitive hyperpolarizing factors, oxygen free radicals, or accumulation of cyclic nucleotides.

Experiments were designed to determine whether normal fluctuations in endogenous sex steroid hormones and/or gender affect endothelium-dependent relaxations in coronary arteries, and, if so, to identify endothelium-derived factors contributing to these differences. Coronary arteries from sexually mature, gonadally intact male and female pigs or ovariectomized pigs were prepared either for study of isometric force in organ chambers or for measurement of prostanoids and activity of nitric oxide (NO) synthase. In organ chamber studies, neither the magnitude nor the sensitivity of endothelium-dependent relaxations correlated with endogenous estrogen or progesterone in female pigs. However, relaxations to bradykinin and UK-14304 were significantly greater and/or shifted leftward in arterial rings from female compared with male pigs. Indomethacin (10(-5) mol/l) increased endothelium-dependent relaxations only in arteries from male pigs. N(G)-monomethyl-L-arginine reduced endothelium-dependent relaxations to a similar extent in coronary arteries from either sex. Neither production nor response to thromboxane A2 or prostacyclin differed in coronary arteries from male compared with female pigs. Activity for calcium-dependent or -independent NO synthase was similar in both sexes. These results suggest that normal fluctuations in endogenous sex steroid hormones do not affect endothelium-dependent relaxations in coronary arteries from female pigs. There are, however, gender differences in endothelium-dependent relaxations that are indomethacin sensitive and may be due to cyclooxygenase products other than thromboxane A2 or prostacyclin.

Background: Adventitial gene transfer may serve as a tool to study vascular biology and may have therapeutic potential. We investigated the hypothesis that adenovirus-mediated transfer of the gene for endothelial nitric oxide synthase (eNOS) to the adventitia would alter vascular reactivity., Methods and Results: Rabbit carotid arteries were surgically isolated and adenoviral vectors encoding eNOS (AdeNOS) or beta-galactosidase instilled into the periarterial sheath at a concentration of 1 x 10(10) pfu/mL. Arteries were harvested 4 days later for immunostaining, NOS enzymatic assay, measurement of cGMP, and vasomotor studies. Transgene expression in the adventitia was confirmed by histochemistry for beta-galactosidase and immunostaining for eNOS with a monoclonal antibody. Calcium-dependent NOS enzymatic activity and cGMP levels were significantly greater in the AdeNOS-transduced arteries. Maximal contractions to phenylephrine (10(-5) mol/L) were diminished in the AdeNOS-transduced arteries (4.6+/-0.2 versus 5.6+/-0.2 g; P<.05), but in the presence of the eNOS inhibitor N(G)-monomethyl-L-argininc (3x10(-4) mol/L) there was no difference between the two groups (7.1+/-0.2 versus 7.5+/-0.3 g; P=NS). Relaxations to calcium ionophore obtained during submaximal contractions to phenylephrine were significantly enhanced in the AdeNOS-transduced arteries (-log EC50, 7.77+/-0.08 versus 7.45+/-0.07; P<.02)., Conclusions: We conclude that eNOS gene transfer to the adventitia alters vascular reactivity, as demonstrated by diminished contractile responses to phenylephrine and enhanced relaxations to calcium ionophore. This may represent a therapeutic strategy for vascular diseases characterized by decreased bioavailability of NO.

Experiments were designed to determine whether normal fluctuations in sex steroid hormones alter gene transcription for endothelial nitric oxide synthase (NOS) and preproendothelin-1 (prepro-ET-1). Aortic endothelial cells were removed from adult, gonadally intact male and female or ovariectomized Yorkshire pigs. Endothelial cells were prepared for Northern blot analysis, Western blot analysis or enzyme activity. Nitric oxide products (NOx) and endothelin-1 (ET-1) in plasma were measured by chemiluminescence and radioimmunoassay, respectively. Northern blot analysis identified single bands corresponding to endothelial NOS and prepro-ET-1. Quantification of the blots showed an increase in expression of mRNA for both endothelial NOS and prepro-ET-1 in ovariectomized pigs compared with gonadally intact male and female pigs. There were no differences in amount of endothelial NOS protein identified by Western blot analysis among groups. On the contrary, plasma concentrations of NOx were significantly decreased in ovariectomized pigs, and there were no differences either in the concentrations of ET-1 in the plasma or extracts from the coronary arteries. These results suggest that expression of endothelial NOS and prepro-ET-1 may be regulated at transcriptional level by ovarian hormones. In addition, the ovarian hormones may regulate production of these endothelium-derived factors at the posttranscriptional level.

Background: Experimental hypercholesterolemia is associated with coronary vasomotor dysfunction. This study was designed to test the hypothesis that experimental hypercholesterolemia is characterized by altered coronary vasomotor responses to endothelin and inhibition of the endogenous NO pathway., Methods and Results: Endothelin-1 (ET-1) at 5 ng x kg(-1) x min(-1) or N(G)-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of nitric oxide synthase (NOS), at 50 microg x kg(-1) x min(-1) was infused into the left anterior descending coronary artery in pigs before and after 10 weeks of cholesterol diet. There was a significant increase in serum cholesterol. At 10 weeks, ET-1 resulted in an accentuated decrease in coronary blood flow (CBF) and coronary artery diameter (CAD) compared with baseline (-88+/-6% versus -45+/-9%, P<.05, and -77+/-14% versus -18+/-8%, P<.05, respectively) and an increase in coronary vascular resistance (CVR) (242+/-18% versus 110+/-17%, P<.05); ET receptor density and binding affinity in epicardial coronary arteries were unchanged. The effect of L-NMMA on CBF, CAD, and CVR was attenuated at 10 weeks (-7+/-8% versus -48+/-4%, -2+/-3% versus -17+/-5%, and 16+/-10% versus 125+/-32%; each P<.05). Immunohistochemistry staining for constitutive NOS revealed a decrease in immunoreactivity in the coronary arteries of hypercholesterolemic pigs., Conclusions: The present study demonstrates an enhanced coronary vasoconstrictive response to pathophysiological doses of endothelin and an attenuated response to the inhibition of endogenous NO activity, suggesting an alteration in coronary vascular reactivity in experimental hypercholesterolemia.

Experiments were designed to characterize endothelin receptors in bronchi and parenchyma of transplanted lungs during acute rejection. Third-order bronchi from autografted or allografted lungs were either cut into rings and suspended in organ chambers for the measurement of isometric force or frozen for isolation of membrane proteins. Lung parenchyma was prepared for histology or isolation of membrane protein. The grade of rejection was 2.74+/-0.17 (n= 19) in allotransplanted lungs; evidence of infection was present in 58% of the transplanted lungs. In organ chamber experiments, endothelin 1 (which stimulates endothelin A receptors) caused comparable contraction of bronchi from autotransplanted and allotransplanted rejecting lungs. Endothelin 3 (which stimulates endothelin A and B receptors) caused contractions of bronchi from autotransplanted lungs which were not different from those caused by endothelin 1. In contrast, contractions caused by endothelin 3 were reduced in bronchi from rejecting allotransplanted lungs. The magnitude of contractions caused by endothelin 3 was reduced further when infection was present with rejection. Competitive inhibition of 125I-endothelin 1 by endothelin 3 was significant for a two-site binding model in membranes prepared from all bronchi and lung parenchyma. The total number of binding sites (Bmax) was reduced significantly in bronchi and parenchyma from rejecting lungs with or without infection. The relative proportions of high-affinity and low-affinity binding sites did not change. Affinities of both high- and low-affinity receptors were not altered with rejection. These results indicate that at least two subtypes of endothelin receptors are present on canine bronchial smooth muscle and parenchyma. The number of endothelin receptors associated with bronchial contractions is reduced with rejection of lung allografts.

Purpose: Experiments were designed to characterize endothelin receptors in human varicose veins. Three groups of veins were studied: (1) varicose vein (VV) tributaries of the greater saphenous vein from patients who were undergoing vein stripping for primary varicosity; (2) greater saphenous veins (SVs) from the same patients; and (3) greater saphenous veins from patients without varicosity who were undergoing arterial reconstruction (control)., Methods: Veins were either cut into rings and suspended in organ chambers for measurement of isometric force, prepared for receptor binding of membrane proteins, or were prepared for measurement of preproendothelin mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR)., Results: Endothelin-1 (10(-11) to 10(-7) mol/L) produced similar concentration-dependent contractions in rings with or without endothelium. Maximal tensions were significantly greater in control veins compared with either SVs or VVs. Sarafotoxin S6c (10(-11) to 3 x 10(-7) mol/L), which is selective for the endothelin-B receptor, also produced concentration-dependent increases in tension in all veins. Sarafotoxin S6c responses in VVs were shifted significantly rightward compared with either SVs or control. Maximal tensions to sarafotoxin S6c also were significantly greater in control veins compared with either SVs or VVs. In receptor binding studies, the number of binding sites as defined by competitive inhibition of 125I-endothelin-1 by endothelin-1 was less in VVs than control veins. Competitive inhibition of 125I-endothelin-1 with endothelin-3 (both A and B receptors) or sarafotoxin S6c (B receptors only) suggests that the difference in receptor number between varicose and nonvaricose veins is attributable to differences in the endothelin-B receptor subtype. Binding affinities were not significantly different for either of the receptor subtypes in all veins studied. Preproendothelin mRNA as quantitated by RT-PCR tended to be higher in VVs compared with either SVs or control veins., Conclusions: Decreased contractions to endothelin-1 in both varicose and saphenous veins of patients with primary varicosity may be associated with a decrease in the number of receptors. These receptors may be downregulated in response to increased production of endothelin-1, which is regulated at the transcriptional level.

Experiments were designed to determine whether endogenous physiological fluctuations in sex steroid hormones affect expression or functional responses of endothelin receptors. Coronary arteries from sexually mature male, female, and ovariectomized pigs were prepared either for receptor binding or measurement of isometric force in organ chambers. Competitive binding of 125I-labeled endothelin-1 was significant for a one-site model with unlabeled endothelin-1 and a two-site model with unlabeled endothelin-3 and sarafotoxin S6c in all pigs. The total number of binding sites for all endothelin ligands was not different between male and female pigs. Binding affinities for the high-affinity binding site for both endothelin-3 and sarafotoxin S6c were significantly greater (lower inhibition constant) in membranes prepared from female pigs with high endogenous estrogen. In organ chamber experiments, contractions to endothelin-1 but not endothelin-3 or sarafotoxin S6c were significantly greater in coronary arterial rings from female compared with male pigs and were not affected significantly by removal of the endothelium or by treatment of the rings with either indomethacin (10(-5) mol/l) or the combination of indomethacin and NG-monomethyl-L-arginine (10(-4) mol/l). These results suggest endogenous fluctuations in estrogen are associated with an increase in affinity of a high-affinity endothelin receptor in coronary arterial smooth muscle of female pigs. In addition, independent of endogenous estrogen status, coronary arteries from female pigs generate significantly greater contractions to endothelin-1 compared with male pigs. This phenomenon occurs at the level of smooth muscle and is not dependent on the endothelium or synthesis of nitric oxide or prostaglandins.

Experiments were designed to determine whether nitric oxide was the mediator of increased endothelium-dependent relaxations in veins proximal to an arteriovenous fistula. A fistula was prepared between femoral arteries and veins in dogs. After 6 wk, veins proximal to the fistula were removed, cut into rings, and suspended for the measurement of isometric force in organ chambers. In some rings the endothelium was removed deliberately. NG-monomethyl-L-arginine (L-NMMA) caused contraction in three of six fistula-operated veins with and without endothelium. In rings contracted submaximally with prostaglandin F2 alpha, acetylcholine and the alpha 2-adrenergic agonist UK-14,304 cause e tylcholine and the alpha 2-adrenergic agonist UK-14,304 caused endothelium-dependent, concentration-dependent relaxations that were greater in fistula compared with sham-operated veins. These relaxations were reduced by L-NMMA. Calcium ionophore A23187 caused comparable endothelium-dependent relaxations in fistula- and sham-operated veins that were unaffected by L-NMMA. There were no differences in either calcium-dependent or -independent activity of nitric oxide synthase isolated from fistula- and sham-operated veins. Positive staining for nitric oxide synthase was present in both the endothelium and media of fistula-operated veins. These results indicate that nitric oxide mediates increased endothelium-dependent relaxations to acetylcholine and alpha 2-adrenergic agonists in fistula-operated veins. Therefore, chronic increases in blood flow and oxygen tension modify selectively receptor-coupled production of nitric oxide in endothelium and smooth muscle of veins.

1. Experiments were designed to determine how sex hormonal status may influence production and response to endothelins. 2. Circulating concentrations of endothelin-1 were not different between sexually mature male and female pigs and did not differ between females of low- and high-oestrogen status. 3. Expression of messenger RNA for preproendothelin was not different in aortic endothelial cells derived from sexually mature male and female pigs. 4. Endothelin-1, endothelin-3 and sarafotoxin S6c caused concentration-dependent contractions of rings of coronary arteries suspended for the measurement of isometric force. Only in response to endothelin-1 were contractions greater in arteries from female than in male pigs. This difference was not related to oestrogen levels in females. 5. The intrinsic force-calcium relationship defined in smooth muscle cells permeabilized with either triton-X or beta-escine was not different in coronary arteries from male and female pigs. The calcium sensitivity was increased to the same extent by endothelin-1 in smooth muscle from male and female pigs. 6. Competitive inhibition of radiolabelled endothelin-1 by unlabelled endothelin-1 was not different in membranes prepared from coronary arteries from male and female pigs. However, competitive inhibition of radiolabelled endothelin-1 by unlabelled endothelin-3 showed two binding sites. The affinity of the high affinity binding site was increased only in females with high oestrogen levels. 7. These results suggest that there are gender differences in contractions of porcine coronary arteries to endothelin-1. These differences may not relate to transcriptional regulation for the production of the peptide in endothelial cells, to the regulation of the number or affinity of endothelin receptors or to intrinsic calcium-sensitivity of the contractile proteins. Rather they are probably due to differences in the regulation of intracellular calcium.

Experiments were designed to characterize endothelin receptors in arteries after chronic increases in blood flow. A fistula was created between the femoral artery and vein in one hindlimb of dogs; contralateral blood vessels were sham operated. Sham- and fistula-operated arteries were removed 6 wk postoperatively. Some arteries were prepared for measurement of isometric force or for isolation of membrane proteins. Other arteries were used for histological staining with an endothelin-B (ETB) receptor antibody. In arteries suspended for the measurement of isometric force, endothelin-1 produced concentration-dependent increases in tension that were significantly greater in fistula- than in sham-operated arteries without endothelium. The ETB-receptor-selective peptide sarafotoxin S6c produced concentration-dependent increases in tension only in fistula-operated arteries. In receptor-binding studies of membrane proteins, Scatchard analysis of saturation binding with 125I-labeled endothelin-1 (125I-endothelin-1) indicated that the total number of receptors was greater in fistula-operated arteries; affinity was threefold less in fistula- than in sham-operated arteries. Competitive displacement of 125I-endothelin-1 by endothelin-3 was significant for a two-site model in membranes prepared from sham-and fistula-operated arteries. Competitive inhibition of 125I-endothelin-1 binding by sarafotoxin S6c was significant for a one-site binding model in all arteries. Sarafotoxin S6c binding sites were elevated significantly in fistula-operated arteries. Immunohistochemical staining for the ETB receptor was significantly greater in both the endothelium and smooth muscle of fistula- than in sham-operated arteries. These results suggest that chronic increases in blood flow upregulate endothelin receptors, including ETB receptors in arterial smooth muscle.

The use of lucigenin-enhanced chemiluminescence (CL) for the detection of superoxide (O2.-) has grown in popularity due to an increased demand for a simple and specific system capable of measuring superoxide. In this study we report a lucigenin-CL signal emanating from human saphenous veins (SV) that was not inhibited by superoxide dismutase (SOD) and lasted for more than 24 h. A larger CL-signal with similar properties was produced by saphenous veins that had been dehydrated. A similar, non-SOD-inhibitable lucigenin-CL was also produced with a variety of phospholipids and phosphatidic acid. The chemical moiety responsible for the phospholipid CL is oxygen dependent but remains unidentified because a variety of lipids and phosphate containing species failed to produce such a signal. These results suggest that the use of lucigenin as a specific CL enhancer for O2.- must be clearly discriminated with a specific O2.- inhibitor when used in biological systems.

Methylene blue is occasionally applied to the adventitia of blood vessels during coronary artery bypass and other vascular procedures to assist in the orientation of the vessel. Inherent in this method is the assumption that extravascular application of methylene blue is innocuous with regard to vascular function. In the first part of this study, the in vitro vascular reactivity of methylene blue-labeled saphenous veins was compared with that of veins that were not marked with methylene blue. The vasoactive agents tested were designed to examine multiple pathways. They included potassium chloride, prostaglandin F2 alpha, phenylephrine, serotonin, angiotensin II, BHT-933 (alpha 2-adrenergic agonist), sodium nitroprusside, acetylcholine, isoproterenol, and verapamil. Compared with unmarked veins, those marked with methylene blue demonstrated a significant impairment of both vasoconstrictor and vasodilator function. These observations were made on a relatively small number of patients and could therefore be attributed to inherent differences between patients or surgical procedures. In the second part of this study, these variables were eliminated by dividing a single vein from one patient into three segments for a 45-minute exposure to external only methylene blue, internal and external methylene blue, or no methylene blue. The segments were then evaluated for vasoreactivity in vitro. Externally applied methylene blue reduced vasoconstriction regardless of the agonist. Further, both endothelium-dependent and -independent vasodilation was diminished by external methylene blue exposure. In veins exposed to methylene blue both internally and externally the results were similar but the magnitude of impairment greater. It is concluded that surgical marking of blood vessels with methylene blue has the potential to adversely affect vascular reactivity and therefore the use of alternative dyes should be considered.

The arcuate nucleus is the bed nucleus for the pro-opiomelancortin system of the brain with important connections with other nuclei involved in cardiovascular function. Clonidine has been reported to produce its cardiovascular effects through an interaction with opioid and alpha 2-adrenergic receptors. The present study examined the arcuate nucleus as a site of action of clonidine. Male spontaneously hypertensive rats were anesthetized with pentobarbital and were instrumented for the measurement of blood pressure and heart rate. Cannulae were placed either through the cisterna magna (IC) or in the arcuate nucleus. Administration of clonidine (0.03-3.75 micrograms, IC) produced a dose-dependent hypotension and bradycardia. Pretreatment with naloxone (30 micrograms, IC) prior to clonidine administration resulted in a significant attenuation of both the clonidine-induced hypotension and bradycardia. In contrast, administration of naloxone (100 ng) into the arcuate nucleus prior to the central administration of clonidine did not alter the cardiovascular effects of clonidine. These results support the role of central opioidergic receptors in the cardiovascular effects of clonidine but do not support the arcuate nucleus as the site of action.

In 1989, nearly 43% of deaths in the United States were due to some form of cardiovascular disease, and 23% were caused by cancer. Thus, two of every three people in this country die from either cardiovascular disease or cancer. Based on both experimental and epidemiological evidence, investigators believe that free radicals play a critical role in the development of both diseases. Low levels of antioxidants, which increases free radical activity, are clearly associated with an increased risk of these diseases. This link has led to the conclusion that use of antioxidant vitamin supplements to scavenge free radicals could potentially decrease the risks of cancer and cardiovascular disease. Results from numerous studies to date have been very promising, although a true protective or preventive causal relationship has not yet been established. Numerous primary and secondary intervention trials currently underway should more definitively assess the role of antioxidants in disease prevention. In the interim, many people feel the evidence is now strong enough to begin supplementation on their own. The pharmacist is in a position to advise patients on the safe and moderate use of antioxidants. The antioxidants discussed in this article are relatively non-toxic, with the exception of vitamin A. The possible benefits of vitamin A are better achieved with the use of beta-carotene. Megadose antioxidant supplementation does not appear to provide any additional benefit beyond what a more moderate supplement can provide and should therefore be discouraged. Taking a trace mineral with antioxidant potential is generally a waste of money, provided the patient is not initially deficient in the element.(ABSTRACT TRUNCATED AT 250 WORDS)

Norcocaine is the N-demethylated metabolite of cocaine. It is present in the CNS and is reported to be pharmacologically active. The present study was designed to evaluate the cardiovascular actions of norcocaine following central administration. Wistar Kyoto (WKY) rats were anesthetized with pentobarbital and instrumented for measurement of blood pressure and renal and hindlimb blood flow (via Doppler flowprobes). A cerebroventricular cannula was placed in the lateral ventricle for drug administration. Cocaine or norcocaine was administered centrally in a dose range of 0.025 to 4.0 mg/kg. Under the above experimental conditions, 4.0 mg/kg of norcocaine decreased blood pressure without a significant change in either hind limb or renal blood flow. Central administration of cocaine also produced a similar depressor response. In conscious, unrestrained rats, cocaine produced a pressor response while norcocaine did not significantly alter blood pressure. The depressor response to both cocaine and norcocaine in the anesthetized animal is speculated to be due to the local anesthetic properties of the drugs.

Topsentin, a bis(indolyl)imidazole marine natural product, inhibited the proliferation of cultured human and murine tumor cells at micromolar concentrations (IC50 values ranged from 4 to 40 microM) and was active against in vivo P388 leukemia (%T/C = 137, 150 mg/kg, QD1-5) and B16 melanoma (%T/C = 144, 37.5 mg/kg, QD1-9) tumors. Effects of 30 microM topsentin (1-hr exposures) on incorporation of radiolabeled precursors by P388 cells indicated inhibition of DNA synthesis (91%) and to a lesser extent RNA synthesis (57%), whereas synthesis of protein was unaffected (0%). Fluorescence spectral changes and competitive binding experiments with ethidium bromide indicated that topsentin interacted with DNA. No evidence for intercalation was observed in DNA unwinding studies, but competitive binding experiments with Hoechst 33342 and CC-1065 indicated that topsentin bound to DNA in the minor groove.

Progesterone receptor-containing T47D human breast cancer cells are responsive to progestins but fail to respond to other steroid hormones, in particular dexamethasone, because they have no measurable levels of receptors for estrogens, androgens, or glucocorticoids. To quantitatively study dual responsiveness of the mouse mammary tumor virus (MMTV) promoter to progestins and glucocorticoids, we have stably transfected T47D cells with a glucocorticoid receptor (GR) expression vector. A cloned derivative (A1-2) was isolated that expresses a normal, full length GR, as assessed by steroid binding and Western immunoblot with a monoclonal anti-GR antibody. Moreover, GR is expressed at levels (80,000-100,000 molecules per cell) comparable to the high levels of endogenous progesterone receptor (200,000 molecules per cell). In A1-2 cells transiently transfected with an MMTV-chloramphenicol acetyl transferase reporter gene, induction by glucocorticoid was substantially greater (5-fold) than induction mediated by progestins. These results suggest that glucocorticoids may be the primary regulator of MMTV.