RESUMEN
There is limited knowledge on the biological relatedness between citizens and on the demographical dynamics within villages, towns and cities in pre-17th century Western Europe. By combining Y-chromosomal genotypes, in-depth genealogies and surname data in a strict genetic genealogical approach, it is possible to provide insights into the genetic diversity and the relatedness between indigenous paternal lineages within a particular community at the time of the surname adoption. To obtain these insights, six Flemish communities were selected in this study based on the differences in geography and historical development. After rigorous selection of appropriate DNA donors, low relatedness between Y chromosomes of different surnames was found within each community, although there is co-occurrence of these surnames in each community since the start of the surname adoption between the 14th and 15th century. Next, the high communal diversity in Y-chromosomal lineages was comparable with the regional diversity across Flanders at that time. Moreover, clinal distributions of particular Y-chromosomal lineages between the communities were observed according to the clinal distributions earlier observed across the Flemish regions and Western Europe. No significant indication for genetic differences between communities with distinct historical development was found in the analysis. These genetic results provide relevant information for studies in historical sciences, archaeology, forensic genetics and genealogy.
Asunto(s)
Cromosomas Humanos Y/genética , Variación Genética , Genética de Población/historia , Nombres , Bélgica , Europa (Continente) , Genotipo , Historia del Siglo XV , Historia Medieval , Humanos , Linaje , Análisis de Secuencia de ADNRESUMEN
Recent evidence suggests that seeking out extra-pair paternity (EPP) can be a viable alternative reproductive strategy for both males and females in many pair-bonded species, including humans. Accurate data on EPP rates in humans, however, are scant and mostly restricted to extant populations. Here, we provide the first large-scale, unbiased genetic study of historical EPP rates in a Western European human population based on combining Y-chromosomal data to infer genetic patrilineages with genealogical and surname data, which reflect known historical presumed paternity. Using two independent methods, we estimate that over the last few centuries, EPP rates in Flanders (Belgium) were only around 12% per generation. This figure is substantially lower than the 830% per generation reported in some behavioural studies on historical EPP rates, but comparable with the rates reported by other genetic studies of contemporary Western European populations. These results suggest that human EPP rates have not changed substantially during the last 400 years in Flanders and imply that legal genealogies rarely differ from the biological ones. This result has significant implications for a diverse set of fields, including human population genetics, historical demography, forensic science and human sociobiology.
Asunto(s)
Cromosomas Humanos Y/genética , Genotipo , Paternidad , Polimorfismo de Nucleótido Simple , Bélgica , Cromosomas Humanos Y/metabolismo , Femenino , Haplotipos , Humanos , Masculino , Datos de Secuencia Molecular , Linaje , Reacción en Cadena en Tiempo Real de la Polimerasa , Reproducción , Factores de TiempoRESUMEN
AIMS: Diabetic polyneuropathy (DPN) without or with neuropathic pain (DPN-P) is one of the most frequent complications of diabetes. To better delineate their respective prevalences, we conducted a cross-sectional study that included 1111 patients (767 type 2 and 344 type 1 diabetic patients) followed up in diabetic outpatients clinics. The association of DPN and DPN-P with other diabetic complications, the impact on quality of life (QoL) and pain management were also investigated. METHODS: Two validated tools (Neuropen) and the DN4 questionnaire) were used to diagnose the two conditions. Pain intensity was measured using a visual analogue scale, and participants completed the 12-item Short-Form Health Survey to evaluate the physical and mental components of QoL. Univariate and multivariate models were used for the statistical analyses. RESULTS: The prevalence of DPN was 43% (95% CI 40.1-45.9), and was higher in type 2 (50.8%) than in type 1 (25.6%) diabetic patients. The prevalence of DPN-P was 14% (95% CI 12.1-16.2) which, again, was higher in type 2 (17.9%) than in type 1 (5.8%) patients. These prevalences both increased with age and diabetes duration. Nephropathy, obesity, low HDL cholesterol and high triglyceride levels were independently associated with DPN and/or DPN-P. Physical and mental components of QoL were significantly altered by DPN-P, but not DPN. Only half of the DPN-P patients were using analgesic treatment, while 28% were using anticonvulsants or antidepressants. CONCLUSION: DPN and DPN-P are frequent complications of diabetes, especially in type 2, and can be identified with inexpensive and easy-to-use screening tools. Despite its profound impact on QoL, DPN-P remains undertreated.
Asunto(s)
Diabetes Mellitus Tipo 1/fisiopatología , Diabetes Mellitus Tipo 2/fisiopatología , Neuropatías Diabéticas/fisiopatología , Pacientes Internos , Pacientes Ambulatorios , Dolor/fisiopatología , Enfermedades del Sistema Nervioso Periférico/fisiopatología , Calidad de Vida , Adulto , Bélgica , Presión Sanguínea , Índice de Masa Corporal , Diabetes Mellitus Tipo 1/psicología , Diabetes Mellitus Tipo 2/psicología , Neuropatías Diabéticas/complicaciones , Neuropatías Diabéticas/psicología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Dolor/etiología , Dolor/psicología , Enfermedades del Sistema Nervioso Periférico/psicologíaRESUMEN
We describe the case of a 71-years-old man in chronic hospital hemodialysis who was admitted to the hospital because of symptomatic hypoglycemia. We discovered that this was due to a documented intoxication with cibenzoline, an antiarrhythmic drug, used to treat (supra-)ventricular tachyarrhythmia. In addition we made a short review of the literature concerning cibenzoline intoxication.
Asunto(s)
Antiarrítmicos/efectos adversos , Fibrilación Atrial/tratamiento farmacológico , Hipoglucemia/inducido químicamente , Imidazoles/efectos adversos , Fallo Renal Crónico/terapia , Diálisis Renal , Anciano , Antiarrítmicos/administración & dosificación , Fibrilación Atrial/epidemiología , Comorbilidad , Humanos , Imidazoles/administración & dosificación , Fallo Renal Crónico/epidemiología , MasculinoRESUMEN
BACKGROUND: The internal cardioverter defibrillator (ICD) is increasingly used to treat ventricular tachyarrhythmias in patients with coronary artery disease (CAD). The burden of coronary risk factors and inflammation is however not well studied in these high risk patients. STUDY AIMS: The aim of the present study was to describe the prevalence of coronary risk factors (including lipid values) and inflammation (including high sensitive-C-reactive protein, hs-CRP) in patients with CAD and ICD implants. METHODS: Baseline clinical characteristics and laboratory results of all eligible patients for the Cholesterol Lowering and Arrhythmias Recurrences after Internal Defibrillator Implantation trial (CLARIDI trial) were used. All patients had documented CAD, an ICD implant and were not yet treated with statins. Coronary risk factors, lipid values, glycated haemoglobin (HbA(1c)) and hs-CRP levels were determined. RESULTS: In the 110 included patients (mean age 68+/-9 years, LVEF 40+/-17%, NYHA class II-III in 47%), a high prevalence of coronary risk factors was documented: current smoking in 18%, body mass index > or =30 kg/m(2) in 16%, blood pressure > or =140/90 mm Hg in 40%, history of diabetes in 12%, and HbA(1c) > or =6% in 16% of patients not known with diabetes. A total cholesterol >175 mg/dl was found in 76% of patients and an LDL cholesterol >100 mg/dl in 83%. Finally, median hs-CRP was 4.8 mg/l (interquartile range 2.5-13.9 mg/l). Hs-CRP values > or =2 mg/l were noted in 83% of all patients and in 68% of patients who had an ICD implant more than 6 months before inclusion. CONCLUSION: In CAD patients with ICD implants, the burden of coronary risk factors is high, often unrecognized and/or under-treated. Persistent inflammation is found in the majority of these patients.
Asunto(s)
Enfermedad de la Arteria Coronaria/sangre , Desfibriladores Implantables , Mediadores de Inflamación/sangre , Inflamación/sangre , Adulto , Anciano , Anticolesterolemiantes/uso terapéutico , Atorvastatina , Bélgica/epidemiología , Biomarcadores/sangre , Proteína C-Reactiva/metabolismo , Enfermedad de la Arteria Coronaria/tratamiento farmacológico , Enfermedad de la Arteria Coronaria/epidemiología , Método Doble Ciego , Femenino , Estudios de Seguimiento , Hemoglobina Glucada/metabolismo , Grecia/epidemiología , Ácidos Heptanoicos/uso terapéutico , Humanos , Lípidos/sangre , Masculino , Persona de Mediana Edad , Péptido Natriurético Encefálico/sangre , Fragmentos de Péptidos/sangre , Prevalencia , Pirroles/uso terapéutico , Proyectos de Investigación , Factores de Riesgo , Taquicardia Ventricular/sangre , Taquicardia Ventricular/terapia , Resultado del Tratamiento , Fibrilación Ventricular/sangre , Fibrilación Ventricular/terapiaRESUMEN
OBJECTIVE: To seek an association between total arterial compliance (TAC) and the extent of ischaemia at stress echocardiography. DESIGN: Cohort study. SETTING: Regional cardiac centre. METHODS: 255 consecutive patients (147 men; mean (SD) age 58 (8)) presenting for stress echocardiography for clinical indications were studied. Wall motion score index (WMSI) was calculated and ischaemia was defined by an inducible or worsening wall motion abnormality. Peak WMSI was used to reflect the extent of dysfunction (ischaemia or scar), and DeltaWMSI was indicative of extent of ischaemia. TAC was assessed at rest by simultaneous radial applanation tonometry and pulsed wave Doppler in all patients. RESULTS: Ischaemia was identified by stress echocardiography in 65 patients (25%). TAC was similar in the groups with negative and positive echocardiograms (1.08 (0.41) v 1.17 (0.51) ml/mm Hg, not significant). However, the extent of dysfunction was associated with TAC independently of age, blood pressure, risk factors, and use of a beta blocker. Moreover, the extent of ischaemia was determined by TAC, risk factors, and use of a beta blocker. CONCLUSION: While traditional cardiovascular risk factors are strong predictors of ischaemia on stress echocardiography, TAC is an independent predictor of the extent of ischaemia.
Asunto(s)
Vasos Coronarios/fisiopatología , Isquemia Miocárdica/etiología , Presión Sanguínea/fisiología , Adaptabilidad , Enfermedad de la Arteria Coronaria/etiología , Enfermedad de la Arteria Coronaria/fisiopatología , Ecocardiografía de Estrés , Femenino , Humanos , Masculino , Persona de Mediana Edad , Isquemia Miocárdica/fisiopatología , Pulso ArterialRESUMEN
BACKGROUND: Patients with end-stage renal disease commonly present with an atherogenic lipid profile characterized by the accumulation of triglyceride-rich, apoprotein B-containing "remnant" lipoproteins, small dense low-density lipoprotein, and low levels of high-density lipoprotein. They are at increased cardiovascular risk and may benefit from drastic lipid-lowering treatment with atorvastatin, a potent, broadacting lipid regulator. This study aims to assess the effects of atorvastatin on the lipid profile in hemodialysis patients, to determine wether atorvastatin is also effective at lowering lipid levels in this particular high-risk subgroup. METHODS: In this randomized, placebo-controlled, double-blind study in hemodialysis patients with hypercholesterolemia (n = 42, mean total cholesterol 243 +/- 33 mg/dl (6.3 +/- 0.8 mmol/l)), the efficacy of 4-weekly increasing doses of atorvastatin (10 - 40 mg daily) was investigated. Lipids and apoproteins were measured in plasma and isolated lipoprotein fractions. RESULTS: Mean total cholesterol and low-density lipoprotein cholesterol progressively decreased with increasing doses of atorvastatin (total cholesterol -33%, low-density lipoprotein cholesterol -43% after 12 weeks), while high-density lipoprotein cholesterol remained unchanged. Plasma levels of apoprotein B and apoprotein E were also significantly reduced by atorvastatin 10 mg, while up-titration to 20 and 40 mg daily provided additional benefits by lowering triglycerides and apoprotein C-III. At week 12, the fraction of small dense low-density lipoprotein was significantly reduced from 23% - 18%, and apoprotein B-containing intermediate-density lipoproteins were no longer detectable. CONCLUSION: In conclusion, atorvastatin not only treated hypercholesterolemia but also favorably affected the uremic lipid profile in patients on hemodialysis. Atorvastatin 4-weekly dose escalation up to 40 mg daily was well-tolerated. Further prospective studies are needed to evaluate the impact of this improved lipid profile on morbidity and mortality.
Asunto(s)
Apolipoproteínas B/sangre , LDL-Colesterol/sangre , Ácidos Heptanoicos/administración & dosificación , Inhibidores de Hidroximetilglutaril-CoA Reductasas/administración & dosificación , Hipercolesterolemia/tratamiento farmacológico , Pirroles/administración & dosificación , Diálisis Renal , Anciano , Apolipoproteínas B/efectos de los fármacos , Atorvastatina , LDL-Colesterol/efectos de los fármacos , Método Doble Ciego , Femenino , Humanos , Hipercolesterolemia/sangre , Hipercolesterolemia/etiología , Fallo Renal Crónico/sangre , Masculino , Persona de Mediana Edad , Factores de Tiempo , Resultado del TratamientoRESUMEN
We tested whether fractional flow reserve (FFR) discriminates between suboptimally and optimally deployed stents. Latex tubes (diameter solidus in circle = 4 mm) with diameter stenosis 40% (n = 3), 50% (n = 3) and 60% (n = 3) were tested in a pulsatile flow system, using water. Measurements were done at baseline (n = 9; FFR/QCA) and after suboptimal (SOD; 3-mm balloon at 8 atm) and optimal (OD; 4 mm balloon at 16 atm) deployment of a 35-mm stent (n = 6; FFR/QCA/IVUS). Varying Q from 150 to 50 ml/min increased FFR by 2-7%. Conversely, at 100 ml/min, FFR increased by only 0.8% from SOD to OD (P < 0.05). Extrapolating data to blood flow, the gain in FFR from SOD to OD is less than 5% for Q = 100 ml/min, while FFR may increase by 15-20% by changes in blood flow from 50 to 150 ml/min. We conclude that IVUS and QCA are more appropriate for the assessment of optimal stent deployment.
Asunto(s)
Velocidad del Flujo Sanguíneo/fisiología , Angiografía Coronaria , Estenosis Coronaria/fisiopatología , Estenosis Coronaria/cirugía , Stents , Ultrasonografía Intervencional , Circulación Coronaria/fisiología , Estenosis Coronaria/diagnóstico , Vasos Coronarios/diagnóstico por imagen , Vasos Coronarios/cirugía , Diseño de Equipo , Humanos , Índice de Severidad de la Enfermedad , Resultado del TratamientoRESUMEN
In humans intimal thickening is aprerequisite of atherosclerosis. Application of a silicone collar around the rabbit carotid artery induces an intimal thickening but in addition it increases the sensitivity to the vasoconstrictor action of serotonin (5-hydroxytryptamine, 5-HT). The 5-HT receptors involved in collar-induced hypersensitivity to 5-HT were investigated using several agonists and antagonists. One week after placement of collars around both carotid arteries of anaesthetized rabbits, rings (2 mm width) from inside (=collar) and outside (=sham) the collars were mounted in organ baths (10 ml) for isometric force measurements at 6 g loading tension. Collared rings were more sensitive to the contractile effect of 5-HT (7.6 fold) and 5-carboxamidotryptamine (31 fold, 5-CT, 5-HT1 agonist) in cumulative concentration response curves. Sumatriptan (5-HT1B/1D agonist) caused concentration-dependent constrictions in collared rings only. Collar placement did not significantly alter pA2 values (Schild regression) or apparent pKb values (non-linear regression) of spiperone and methysergide (mixed 5-HT2A/5-HT1 antagonists) or ketanserin and ritanserin (5-HT2A antagonists), indicating unchanged binding characteristics of the 5-HT2A receptor. However, the reduced slope of the Schild regression pointed to a heterogeneous receptor population in collared rings. In contrast, the apparent pKb value of methiothepin (5-HT1B antagonist) was significantly reduced by collar placement, and its antagonism shifted from non-surmountable in sham rings to surmountable in collared segments. Taken together, this study demonstrates that the serotonergic receptor involved in the hypersensitivity to 5-HT of rabbit collared carotid artery is a 5-HT1B receptor subtype.
Asunto(s)
Tirantes/efectos adversos , Arterias Carótidas/efectos de los fármacos , Hipersensibilidad/etiología , Receptores de Serotonina/fisiología , Serotonina/farmacología , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Animales , Arterias Carótidas/fisiopatología , Relación Dosis-Respuesta a Droga , Técnicas In Vitro , Ketanserina/farmacología , Masculino , Metisergida/farmacología , Oxadiazoles/farmacología , Piperazinas/farmacología , Cloruro de Potasio/farmacología , Conejos , Receptor de Serotonina 5-HT1B , Receptor de Serotonina 5-HT2A , Receptores de Serotonina/efectos de los fármacos , Ritanserina/farmacología , Serotonina/análogos & derivados , Antagonistas de la Serotonina/farmacología , Agonistas de Receptores de Serotonina/farmacología , Espiperona/farmacología , Sumatriptán/farmacología , Triptaminas/farmacología , Vasoconstricción/efectos de los fármacosRESUMEN
Many studies have shown that loss of endothelium-derived nitric oxide is a major factor of ischemic episodes in patients with coronary artery disease and there is increasing evidence to suggest that nitric oxide might exert antiatherosclerotic actions. Based on these concepts, the results of animal studies on the effects of lipid lowering drugs, antioxidants, angiotensin converting enzyme inhibitors, Ca2+ channel blockers, estrogens and agents which modulate nitric oxide bioavailability are presented and compared to the results of patient studies and clinical trials. In spite of encouraging results obtained with antioxidants in animals, clinical trials could only show a clear positive effect of vitamin E treatment on the outcome of cardiovascular disease. Angiotensin converting enzyme inhibitors can ameliorate endothelial dysfunction in coronary heart disease, but their impact on disease progression remains unclear. There is evidence that estrogen replacement therapy in post-menopausal women may increase the bioavailability of nitric oxide. Finally, improved endothelial function and plaque stability clearly contribute to the clinical benefits of lipid lowering interventions, statins in particular. Taken together, these studies lend support to the concept that improving endothelial function and nitric oxide release might serve as valuable elements in the prevention or therapy of cardiovascular disease.
Asunto(s)
Antioxidantes/farmacología , Arteriosclerosis/prevención & control , Enfermedades Cardiovasculares/tratamiento farmacológico , Enfermedades Cardiovasculares/prevención & control , Endotelio Vascular/metabolismo , Óxido Nítrico/farmacología , Animales , Anticolesterolemiantes/farmacología , Arteriosclerosis/etiología , Ensayos Clínicos como Asunto , HumanosRESUMEN
Oxidized low density lipoprotein (oxLDL) is present in atherosclerotic lesions and has been implicated in the etiopathogenesis of atherosclerosis mainly based on in vitro studies. In view of the lack of data on the activity of oxLDL in vivo, we decided to study its effects in the rabbit by local application at the level of the vascular wall. Intimal thickening was evoked by the placement of a silicone collar around the carotid arteries during 2 weeks. The collar was connected to an osmotic minipump containing human oxLDL (7 micrograms h-1), LDL (7 micrograms h-1) or phosphate-buffered saline. Collar placement resulted in a thickening of the intima thereby increasing the thickness from 5 +/- 1 to 26 +/- 5 microns with the appearance of alpha-actine positive smooth muscle cells. Perivascular infusion of LDL or oxLDL significantly enhanced the intima, containing large amounts of T-lymphocytes, collagen and smooth muscle cells. The placement of the collar and the infusion of oxLDL during 14 days resulted in an increased sensitivity to serotonin and a decreased sensitivity to acetylcholine. The maximal relaxation to acetylcholine was reduced by 50% whereas the endothelium-independent relaxation to nitroglycerin were not affected. These results show for the first time that the local application of oxLDL in vivo promotes intimal thickening and impairs the endothelium-dependent relaxations thereby supporting the suggestion that oxLDL plays an important role in the morphological and functional changes present in atherosclerotic blood vessels.
Asunto(s)
Lipoproteínas LDL/metabolismo , Túnica Íntima/metabolismo , Sistema Vasomotor/fisiología , Animales , Humanos , Oxidación-Reducción , ConejosRESUMEN
OBJECTIVES: Based on in vitro studies, oxidized low-density lipoprotein (oxLDL) has been implicated in atherogenesis and the associated deficiency in endothelium-dependent relaxation. The aim of this study was to investigate the effects of in vivo exposure to oxLDL on intimal thickening and relaxing behaviour. METHODS: Intimal thickening was evoked by the placement of silicone collars around the carotid arteries of the rabbit for 3 or 14 days. OxLDL (Cu(2+)-oxidized, 7 micron/h) or the vehicle phosphate-buffered saline (PBS) was infused in the collars via subdermally implanted osmotic minipumps. RESULTS: The collared vessels receiving PBS developed discrete intimal thickening after 14 days (intima/media (I/M) ratio 11 +/- 2%). OxLDL infusion resulted in intimal thickening after 3 days and significantly enhanced the intimal thickness by 14 days (I/M ratio 98 +/- 16%). Collaring alone for 3 or 14 days and 3 days exposure to oxLDL did not impair the endothelium-dependent relaxations to acetylcholine or calcium ionophore, nor to the NO donors glyceryl trinitrate (GTN) and S-nitroso-N-acetylpenicillamine (SNAP). However, the sensitivity to acetylcholine was decreased after exposure to oxLDL for 14 days (-logEC50 oxLDL 6.95 +/- 0.11 vs. 7.52 +/- 0.11 collar alone) and the maximal relaxation to the endothelium-dependent agonist was reduced by 50%, this in the presence of a virtually intact endothelium. Complete relaxation was still obtained with the nitric oxide donors. CONCLUSION: Our results show for the first time that local vascular exposure to oxLDL in vivo promotes intimal thickening and inhibits endothelium-dependent dilation, thereby supporting an active role for oxLDL in the morphological and functional changes observed in atherosclerotic blood vessels.
Asunto(s)
Endotelio Vascular/efectos de los fármacos , Lipoproteínas LDL/farmacología , Túnica Íntima/efectos de los fármacos , Vasodilatación/efectos de los fármacos , Acetilcolina/farmacología , Animales , Calcimicina/farmacología , Cobre/farmacología , Relación Dosis-Respuesta a Droga , Ionóforos/farmacología , Peroxidación de Lípido , Lipoproteínas LDL/metabolismo , Masculino , Nitroglicerina/farmacología , Penicilamina/análogos & derivados , Penicilamina/farmacología , Fenilefrina/farmacología , Conejos , S-Nitroso-N-Acetilpenicilamina , Túnica Íntima/patología , Vasoconstrictores/farmacología , Vasodilatadores/farmacologíaRESUMEN
Oxidized LDL (oxLDL) has been implicated in atherogenesis on the basis of in vitro studies and is present in atherosclerotic lesions. The aim of this study was to investigate the effects of LDL and oxLDL on intimal thickening in vivo. Intimal thickening was evoked by the placement of silicone collars around the carotid arteries of rabbits for 2 weeks. The collars were connected to osmotic minipumps containing LDL (7 micrograms h-1, n = 16 arteries), oxLDL (Cu2+ oxidized, 7 micrograms h-1, n = 16), or phosphate-buffered saline (5 microL h-1, n = 16). Segments proximal to the collars served as controls. Collar placement without lipoprotein application resulted in the appearance of alpha-SMC actin-immunoreactive cells in the intima, thereby increasing the intimal thickness from 5 +/- 1 to 26 +/- 5 microns. The perivascular infusion of LDL or oxLDL within the collar significantly enhanced the development of the intima ninefold and sevenfold, respectively. The large intimas resulting from lipoprotein exposure were infiltrated by macrophages and T lymphocytes, and the intimal collagen area was increased from 5 +/- 2% in the discrete collar-induced intima to approximately 20% in the lipoprotein-evoked lesions. In conclusion, the local vascular application of LDL, oxidized in vitro or possibly in vivo, elicited an inflammatory-fibroproliferative response characteristic of arteriosclerotic lesions, thereby demonstrating an active role for this class of lipoproteins in the disease process.
Asunto(s)
Arterias Carótidas/efectos de los fármacos , Estenosis Carotídea/patología , Lipoproteínas LDL/toxicidad , Túnica Íntima/efectos de los fármacos , Animales , Arterias Carótidas/patología , Estenosis Carotídea/etiología , Colágeno/análisis , Constricción , Humanos , Hiperplasia , Bombas de Infusión Implantables , Peroxidación de Lípido , Lipoproteínas LDL/farmacología , Masculino , Músculo Liso Vascular/patología , Conejos , Sustancias Reactivas al Ácido Tiobarbitúrico/análisis , Túnica Íntima/química , Túnica Íntima/patologíaRESUMEN
Nitric oxide (NO) plays an important role in the cytotoxic activity of macrophages towards tumour cells and microbial pathogens. We investigated whether alteration of intracellular thiol levels modulates the cytotoxic effects of different NO donors and lipopolysaccharide-induced NO in the murine macrophage cell lin J774A.1. The NO-releasing compound S-nitroso-N-acetylpenicillamine caused a significant concentration-dependent loss of viability of the macrophages only under glucose-limiting conditions. The cytotoxic effect of S-nitroso-N-acetylpenicillamine was prevented by the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO). Depletion of total glutathione before exposure to S-nitroso-N-acetylpenicillamine further decrease cell viability while pretreatment with N-acetylcysteine was protective. Comparing equimolar concentrations of various NO donors including S-nitrosoglutathione, S-nitrosocysteine and 3-morpholino-sydnonimine hydrochloride, cytotoxicity appeared to be related to the relative stability of the test compound. Both the order of stability and the order of potency for cell killing was S-nitrosoglutathione > S-nitroso-N-acetylpenicillamine > S-nitrosocysteine = 3-morpholino-sydnonimine hydrochloride. Stimulation of the macrophages with lipopolysaccharide and interferon-gamma resulted in dose-dependent cell injury and NO production. Glutathione depletion prior to stimulation considerably decreased macrophage viability as well as the NO production. In contrast to the protective effect on S-nitroso-N-acetylpenicillamine-mediated injury, pretreatment with N-acetylcysteine did not influence the lipopolysaccharide-mediated cytotoxicity. These results demonstrate that (a) reduction in the availability of glucose and intracellular glutathione renders the cells more vulnerable to the cytotoxic effects of NO donors, (b) in this model of cytotoxicity, long-lived NO donors were more cytotoxic than short-lived NO donors, (c) the differential effects of N-acetylcysteine on S-nitroso-N-acetylpenicillamine-induced and bacterial lipopolysaccharide-mediated cytotoxicity support the existence of other toxic species different from NO or NO-related compounds with a potent cytotoxic activity in immunostimulated macrophages, and (d) other non-protein thiols like N-acetylcysteine may substitute for glutathione as a major component of the cellular antioxidant defense system.
Asunto(s)
Macrófagos/efectos de los fármacos , Óxido Nítrico/antagonistas & inhibidores , Compuestos de Sulfhidrilo/farmacología , Acetilcisteína/farmacología , Animales , Benzoatos/farmacología , Butionina Sulfoximina/farmacología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Inhibidores Enzimáticos/toxicidad , Glutatión/metabolismo , Glutatión/fisiología , Hidroxilamina , Hidroxilaminas/farmacología , Imidazoles/farmacología , Interferón gamma/toxicidad , Lipopolisacáridos/toxicidad , Macrófagos/metabolismo , Ratones , Óxido Nítrico/toxicidad , Penicilamina/análogos & derivados , Penicilamina/antagonistas & inhibidores , Penicilamina/toxicidad , S-Nitroso-N-Acetilpenicilamina , ATPasa Intercambiadora de Sodio-Potasio/antagonistas & inhibidoresRESUMEN
Atherosclerosis is a chronic inflammatory process in the intima of conduit arteries, which disturbs the endothelium-dependent regulation of the vascular tone by the labile liposoluble radical nitric oxide (NO) formed by the constitutive endothelial nitric oxide synthase (eNOS). This defect predisposes to coronary vasospasm and cardiac ischaemia, with anginal pain as the typical clinical manifestation. It is now appreciated that endothelial dysfunction is an early event in atherogenesis and that it may also involve the microcirculation, in which atherosclerotic lesions do not develop. On the other hand, the inflammatory environment in atherosclerotic plaques may result in the expression of the inducible NO synthase (iNOS) isozyme. Whether the dysfunction in endothelial NO production is causal to, or the result of, atherosclerotic lesion formation is still highly debated. Most evidence supports the hypothesis that constitutive endothelial NO release protects against atherogenesis e.g. by preventing smooth muscle cell proliferation and leukocyte adhesion. Nitric oxide generated by the inducible isozyme may be beneficial by replacing the failing endothelial production but excessive release may damage the vascular wall cells, especially in combination with reactive oxygen intermediates.
RESUMEN
Oxidized low-density lipoprotein (LDL) is currently regarded as a tentative key player in atherosclerosis by virtue of its ability to induce intracellular lipid accumulation and to modulate cell functions in the vessel wall. We previously demonstrated that inducible nitric oxide (NO) synthase activity is attenuated in lipid-laden J774 macrophages obtained by incubation with oxidized LDL 200 micrograms ml-1 for 24 h. In the present study we investigated the effect of oxidized LDL in a lower concentration (20 micrograms ml-1) or for a shorter time (6 h) and the possible mediator role of prostaglandin E2 and prostacyclin. Prostaglandins and the NO synthase metabolites citrulline and nitrite were elevated in the 24 h supernatant after immune stimulation with interferon-gamma 100 U ml-1 with or without lipopolysaccharide 10 micrograms ml-1. Pretreatment with oxidized LDL 20 micrograms ml-1 for 18 h decreased nitrite release by 31 +/- 2%, whereas prostaglandin production was not affected. A 6 h pre-exposure to 200 micrograms ml-1 had an opposite effect: it significantly potentiated interferon-gamma-stimulated prostaglandin E2 (10-fold), prostacyclin (7-fold), nitrite (1.5-fold), and citrulline (2.4-fold) release. Indomethacin 10 microM abolished the prostaglandin production and largely prevented the oxidized LDL-dependent increase in NO synthase activity. Acetylated LDL was without effect. The data show that the immune-induced release of NO is potentiated or suppressed, depending on the conditions of exposure to oxidized LDL. The potentiation due to short, high-dose exposure is partly mediated by prostaglandins since indomethacin inhibited both processes.
Asunto(s)
Lipoproteínas LDL/farmacología , Macrófagos/metabolismo , Óxido Nítrico/biosíntesis , Prostaglandinas/biosíntesis , Animales , Células Cultivadas , Citrulina/metabolismo , Indometacina/farmacología , Interferón gamma/farmacología , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Ratones , Óxido Nítrico Sintasa/metabolismo , Nitritos/metabolismo , Oxidación-Reducción , Prostaglandinas/farmacologíaRESUMEN
L-Arginine is converted to the highly reactive and unstable nitric oxide (NO) and L-citrulline by an enzyme named nitric oxide synthase (NOS). NO decomposes into other nitrogen oxides such as nitrite (NO(2) (-)) and nitrate (NO(2) (-)), and in the presence of superoxide anion to the potent oxidizing agent peroxynitrite (ONOO(-)). Activated rodent macrophages are capable of expressing an inducible form of this enzyme (iNOS) in response to appropriate stimuli, i.e., lipopolysaccharide (LPS) and interferon-gamma (IFNgamma). Other cytokines can modulate the induction of NO biosynthesis in macrophages. NO is a major effector molecule of the anti-microbial and cytotoxic activity of rodent macrophages against certain micro-organisms and tumour cells, respectively. The NO synthesizing pathway has been demonstrated in human monocytes and other cells, but its role in host defence seems to be accessory. A delicate functional balance between microbial stimuli, host-derived cytokines and hormones in the microenvironment regulates iNOS expression. This review will focus mainly on the known and proposed mechanisms of the regulation of iNOS induction, and on agents that can modulate NO release once the active enzyme has been expressed in the macrophage.
RESUMEN
Activated cholesterol-laden macrophages in atherosclerotic lesions are believed to influence the progression of this disease. The induction of nitric oxide synthase (iNOS) activity was investigated in control and cholesterol-laden J774 macrophages, obtained by pre-incubation with oxidized or acetylated low density lipoproteins (oxLDL, acLDL). Loading with oxLDL caused a small induction of NOS activity in unstimulated cells, as indicated by nitrite and citrulline accumulation in the supernatant. However, it suppressed the iNOS activity resulting from stimulation of the cells with lipopolysaccharide with or without interferon-gamma. AcLDL had no inhibitory effect, indicating that cholesterol accumulation as such was not responsible. Since the induction of NOS in macrophages is inhibited by glucocorticoids, the possibility that a glucocorticoid-like factor, formed during oxidation of LDL, may cause the inhibition, was investigated. However, addition of the glucocorticoid receptor antagonist mifepristone did not prevent the oxLDL-dependent NOS inhibition, indicating that the glucocorticoid receptor is not involved in the suppressive effect of oxLDL.
RESUMEN
Swan-Ganz semifloating balloon-tipped catheters were introduced in 6 endurance-trained subjects, 7 untrained volunteers, 29 patients suffering from coronary heart disease, 8 patients with right heart insufficiency induced by chronic obstructive syndromes, and 8 patients with idiopathic congestive cardiomyopathy. All subjects except the patients with resting cardiac insufficiency performed graded ergometric tests during the catheter investigation. Hemodynamic values, plasma noradrenaline and adrenaline (in all subjects), and beta-adrenergic receptor density on intact polymorphonuclear leucocytes (in 6 endurance-trained subjects, 5 untrained healthy volunteers, and 6 patients with left heart insufficiency) were determined. For all subjects investigated, significant correlations were observed between resting plasma catecholamine levels and resting hemodynamic values, such as stroke volume (r = 0.47, p less than 0.001), cardiac output (r = 0.32, p less than 0.05), heart rate (r = 0.37, p less than 0.01), pulmonary vascular resistance (r = 0.52, p less than 0.001), and total vascular resistance (r = 0.40, p less than 0.01). An inverse relationship existed between the resting catecholamine levels and the performance ability. Catecholamine levels were approximately three- to fourfold higher and the beta-adrenergic receptor density approximately two- to threefold lower in patients with left heart insufficiency than in healthy untrained subjects. The plasma catecholamine levels were lower and the beta-adrenergic receptor density approximately 60% greater in trained subjects. The beta-adrenergic receptor density may be a critical dynamic parameter for the modulation of sympathetic effects.