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1.
Biologicals ; 45: 85-92, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27756679

RESUMEN

Maternally Derived Antibodies (MDA) can have a negative effect on the efficacy of live attenuated vaccines against classical swine fever (CSF). For this reason, a marker vaccine candidate CP7_E2alf was tested for its efficacy in the presence of MDA. Pregnant sows were vaccinated four weeks before farrowing with CSF virus (CSFV) strain "Thiverval". A total of 40 piglets with MDAs were included in this study. At six weeks of age the piglets were allocated into three treatment groups using generalized randomized block design (GRBD) blocking on serological status and pen location. Of the 40 piglets with MDAs, 30 piglets were vaccinated either orally (n = 15) or intramuscularly (n = 15) with a single dose of vaccine candidate produced under Good Laboratory Practice (GLP) conditions. The ten remaining piglets were allocated into the untreated control group. All 40 piglets were oronasally challenged with 2 ml of the highly virulent CSFV strain "Koslov" 14 days after vaccination. It was revealed that presence of MDAs negatively influences the efficacy of the live marker vaccine candidate, however, the extent of this negative impact depends on the route of vaccine administration. Based on our observations, intramuscular vaccination is recommended during CSF control programs in order to develop superior immune protection.


Asunto(s)
Peste Porcina Clásica , Intercambio Materno-Fetal/inmunología , Porcinos/inmunología , Vacunas Virales/farmacología , Animales , Biomarcadores , Peste Porcina Clásica/inmunología , Peste Porcina Clásica/prevención & control , Femenino , Masculino , Embarazo , Vacunas Virales/inmunología
2.
Biologicals ; 43(2): 92-9, 2015 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-25637578

RESUMEN

Classical swine fever (CSF) marker vaccine candidate CP7_E2alf produced under Good Laboratory Practice (GLP) conditions by Pfizer was tested on 40 six-week-old MDA-piglets according to the European Pharmacopoeia (Ph.Eur.) requirements. Single doses of CP7_E2alf were given to 15 piglets orally, while 15 other piglets were intramuscularly vaccinated. Ten additional animals were included as unvaccinated controls. All piglets were oronasally challenged with the highly virulent CSF virus (CSFV) strain "Koslov" 14 days after vaccination. CP7_E2alf administered i.m. provided a complete protection, while p.o. administratrion triggered only partial protection. The level of protection was determined by the development of clinical signs, viraemia and rate of mortality. The vaccine candidate met the criteria of Ph. Eur Monograph 0065, "Swine-fever vaccine (live, prepared in cell cultures), classical" 7th Edition, which claims the efficacy test is invalid if fewer than 50 per cent of the control piglets display typical signs of serious infection of CSF or die, and if fewer than 100 per cent of the control piglets show clinical signs of disease within 21 days following challenge. Fulfilling these validity criteria is a key step in the registration procedure for a vaccine candidate to become openly available.


Asunto(s)
Virus de la Fiebre Porcina Clásica/inmunología , Peste Porcina Clásica , Vacunas Virales , Animales , Anticuerpos Antivirales , Biomarcadores , Peste Porcina Clásica/inmunología , Peste Porcina Clásica/prevención & control , Femenino , Masculino , Porcinos , Vacunas Virales/inmunología , Vacunas Virales/farmacología
3.
Acta Physiol Hung ; 100(4): 378-87, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24013937

RESUMEN

The aim of our study was to investigate changes in psycho-physiological parameters evoked by short duration, intensive physical stress on university students practicing judo at different intensities and timely manner. Stability of posture, muscle strength (hand force exertions), attention concentration (choice reaction time), cardiac parameters, (ECG, heart rate, heart rate variability), and oxygen saturation were measured, cardiac state and stress index were computed before and after the physical stress. The actual psychic state of the subjects was evaluated using the Spielberger's STPI-H Y-1 test which determined anxiety, curiosity, anger and depression level. Analysis of psychometric and physiologic parameters indicated significant correlations, among others, between force and cardiac stress (-), force and depression (-), anxiety and errors in actions (+), cardiac state and errors in action (-), cardiac state and depression (-). Paired samples tests showed the influence of intensive physical stress within groups of students, and independent samples tests made it possible to evaluate the power of medical and sport students, performing physical training at a significantly higher level than it is usual among the medical students. Our results proved that higher level physical training influences the psychic state advantageously, limits increases in cardiac stress level, and decreases susceptibility to anxiety and depression.


Asunto(s)
Adaptación Psicológica/fisiología , Ansiedad/fisiopatología , Depresión/fisiopatología , Actividad Motora/fisiología , Estrés Fisiológico/fisiología , Adolescente , Femenino , Corazón/fisiología , Humanos , Masculino , Fuerza Muscular/fisiología , Psicofisiología , Tiempo de Reacción/fisiología , Adulto Joven
4.
Acta Vet Hung ; 51(4): 539-50, 2003.
Artículo en Inglés | MEDLINE | ID: mdl-14680066

RESUMEN

The healing process of telescopic anastomoses was found in an animal experiment with 12 mongrel dogs. After the division of vessels an ileal segment of different length was invaginated into the lumen of the colon using single-layer interrupted sutures. The following four groups were used: Group A (n = 3): end-to-side ileocolostomy, single-layer interrupted suture (invagination length: 0 mm), survival time: 21 days. Group B (n = 3): invagination length: 20 mm, survival time: 7 days. Group C (n = 3): invagination length: 10 mm, survival time: 21 days. Group D (n = 3): invagination length: 20 mm, survival time: 21 days. At the end of the above survival times the anastomosis area was removed. The bursting pressure was measured and morphological as well as histological examinations were performed. In each case the 0-day look-alikes of anastomoses were performed using the remnant bowels, and bursting pressure measurements were done on these models as well. Anastomosis leakage did not occur. The serosal layer of the intracolonic part of the ileum disappeared during the healing process. The free surface of the intracolonic ileal segment became covered by the sliding mucosa of the colon and the prolapsing mucosa of the ileum. The following could be concluded after the experiments: The inner pressure tolerance of a telescopic ileocolostomy promptly after preparation is better than in case of another single-layer anastomosis. This fact results in increased safety against leakage on the first postoperative days. The inner pressure tolerance of the telescopic ileocolostomy increases during the healing process and it does not depend on the length of the invaginated part (0 day-20 mm: 56 mmHg +/- 6, Group A: 252 +/- 39, Group B: 154 +/- 19, Group C: 249 +/- 20, Group D: 298 +/- 2). There is no difference in pressure tolerance between the telescopic and the end-to-side single-layer interrupted anastomoses after the healing process. The invaginated section within the lumen of the large intestine does not suffer ischaemic or any other kind of damage. This inexpensive and simple anastomosis technique could be useful in the veterinary surgical practice as well.


Asunto(s)
Anastomosis Quirúrgica/métodos , Colon/cirugía , Íleon/cirugía , Cicatrización de Heridas , Animales , Procedimientos Quirúrgicos del Sistema Digestivo/métodos , Perros
5.
Dis Esophagus ; 16(3): 229-35, 2003.
Artículo en Inglés | MEDLINE | ID: mdl-14641315

RESUMEN

The basis of telescopic anastomosis is old, only the practical details of it have changed and improved. The telescopic anastomosis technique is successfully applied in our practice for reconstruction of gastrectomy and esophageal resection. The reason for this study was that data about the healing process of telescopic anastomosis had not been found in the literature. We used four groups of mongrel dogs for our experiments: Group A (n = 3) received 20 mm-long invaginations with a survival time of 7 days; Group B (n = 3) received 10 mm-long invaginations with a survival time of 21 days; Group C (n = 3) received 20 mm-long invaginations with a survival time of 21 days; Group D (n = 3) received 30 mm-long invaginations with a survival time of 21 days. At the end of the above survival times we removed the anastomosing area, measured the bursting pressures and performed morphological and histological examinations. In each case we also performed an anastomosis exactly the same as a completely healed anastomosis and its pressure tolerance was measured (0 day). The pressure tolerance within the anastomosis rises gradually and independently of the length of the invaginated esophageal part. Anastomosis leakage did not occur. The invaginated esophageal part did not suffer any damage. The muscular wall of the intragastric part of the esophagus became covered by the mucosa of the stomach during the healing process and it joined with the esophageal mucosa at the edge of the free end of the esophagus.


Asunto(s)
Anastomosis Quirúrgica/métodos , Esófago/cirugía , Animales , Perros , Esófago/patología , Esófago/fisiología , Presión , Cicatrización de Heridas
6.
Acta Microbiol Immunol Hung ; 49(2-3): 215-26, 2002.
Artículo en Inglés | MEDLINE | ID: mdl-12109151

RESUMEN

Hungary is facing to perform intensive afforestation. New forests will be planted on dry, poorly fertile soils nonprofitable for agricultural use. Applying artificially mycorrhized seedlings may considerably increase the effectivity of afforestation and decrease costs.


Asunto(s)
Conservación de los Recursos Naturales , Micorrizas/fisiología , Árboles/crecimiento & desarrollo , Árboles/microbiología , Ecosistema , Hungría , Simbiosis
7.
Biochem J ; 359(Pt 2): 315-23, 2001 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-11583577

RESUMEN

The side chain of residue Arg(238) in morphinone reductase (MR) is located close to the N-1/C-2 carbonyl region of the flavin isoalloxazine ring. During enzyme reduction negative charge develops in this region of the flavin. The positioning of a positively charged side chain in the N-1/C-2 carbonyl region of protein-bound flavin is common to many flavoprotein enzymes. To assess the contribution made by Arg(238) in stabilizing the reduced flavin in MR we isolated three mutant forms of the enzyme in which the position of the positively charged side chain was retracted from the N-1/C-2 carbonyl region (Arg(238)-->Lys), the positive charge was removed (Arg(238)-->Met) or the charge was reversed (Arg(238)-->Glu). Each mutant enzyme retains flavin in its active site. Potentiometric studies of the flavin in the wild-type and mutant forms of MR indicate that the flavin semiquinone is not populated to any appreciable extent. Reduction of the flavin in each enzyme is best described by a single Nernst function, and the values of the midpoint reduction potentials (E(12)) for each enzyme fall within the region of -247+/-10 mV. Stopped-flow studies of NADH binding to wild-type and mutant MR enzymes reveal differences in the kinetics of formation and decay of an enzyme-NADH charge-transfer complex, reflecting small perturbations in active-site geometry. Reduced rates of hydride transfer in the mutant enzymes are attributed to altered geometrical alignment of the nicotinamide coenzyme with FMN rather than major perturbations in reduction potential, and this is supported by an observed entropy-enthalpy compensation effect on the hydride transfer reaction throughout the series of enzymes. The data indicate, in contrast with dogma, that the presence of a positively charged side chain close to the N-1/C-2 carbonyl region of the flavin in MR is not required to stabilize the reduced flavin. This finding may have general implications for flavoenzyme catalysis, since it has generally been assumed that positive charge in this region has a stabilizing effect on the reduced form of flavin.


Asunto(s)
Proteínas Bacterianas , Oxidorreductasas/química , Oxidorreductasas/metabolismo , Secuencia de Bases , Dominio Catalítico/genética , Cartilla de ADN/genética , Electroquímica , Estabilidad de Enzimas , Flavinas/química , Flavinas/metabolismo , Cinética , Mutagénesis Sitio-Dirigida , NAD/química , NAD/metabolismo , Oxidación-Reducción , Oxidorreductasas/genética , Potenciometría , Pseudomonas putida/enzimología , Pseudomonas putida/genética
8.
J Mol Biol ; 310(2): 433-47, 2001 Jul 06.
Artículo en Inglés | MEDLINE | ID: mdl-11428899

RESUMEN

Pentaerythritol tetranitrate reductase (PETN reductase) degrades high explosive molecules including nitrate esters, nitroaromatics and cyclic triazine compounds. The enzyme also binds a variety of cyclic enones, including steroids; some steroids act as substrates whilst others are inhibitors. Understanding the basis of reactivity with cyclic enones requires structural information for the enzyme and key complexes formed with steroid substrates and inhibitors. The crystal structure of oxidised and reduced PETN reductase at 1.5 A resolution establishes a close structural similarity to the beta/alpha-barrel flavoenzyme, old yellow enzyme. In complexes of oxidised PETN reductase with progesterone (an inhibitor), 1,4-androstadiene-3,17-dione and prednisone (both substrates) the steroids are stacked over the si-face of the flavin in an orientation different from that reported for old yellow enzyme. The specifically reducible 1,2 unsaturated bonds in 1,4-androstadiene-3,17-dione and prednisone are not optimally aligned with the flavin N5 in oxidised enzyme complexes. These structures suggest either relative "flipping" or shifting of the steroid with respect to the flavin when bound in different redox forms of the enzyme. Deuterium transfer from nicotinamide coenzyme to 1,4-androstadiene-3,17-dione via the enzyme bound FMN indicates 1alpha addition at the steroid C2 atom. These studies rule out lateral motion of the steroid and indicate that the steroid orientation is "flipped" in different redox states of the enzyme.


Asunto(s)
Oxidorreductasas/química , Oxidorreductasas/metabolismo , Esteroides/metabolismo , Androstadienos/química , Androstadienos/metabolismo , Sitios de Unión , Cristalografía por Rayos X , Deuterio , Mononucleótido de Flavina/metabolismo , Ligandos , Espectroscopía de Resonancia Magnética , Modelos Moleculares , NADPH Deshidrogenasa/química , Oxidación-Reducción , Oxidorreductasas/antagonistas & inhibidores , Prednisona/química , Prednisona/metabolismo , Progesterona/química , Progesterona/metabolismo , Unión Proteica , Estructura Secundaria de Proteína , Protones , Soluciones , Estereoisomerismo , Esteroides/química
9.
Pediatr Res ; 49(6): 826-33, 2001 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-11385145

RESUMEN

Angiotensin II (ANG II) contracts umbilical arteries and has been hypothesized to regulate fetal blood pressure primarily by altering umbilical vascular resistance. To determine whether systemic arteries in term fetal sheep are sensitive to ANG II, isometric contraction of endothelium-intact isolated fetal renal, mesenteric, and umbilical arteries in response to ANG II was studied. ANG II (10(-7) M) elicited contractile responses in all three vessels (43 +/- 8%, 99 +/- 21%, and 105 +/- 5% of the maximal response seen with 90 mM KCl for renal, mesenteric, and umbilical arteries, respectively). The time course of the contractile responses differed among the vessels: renal and mesenteric arteries exhibited rapid transient contraction followed by relaxation, whereas umbilical artery displayed a more slowly developing but sustained contraction (1 +/- 0%, 3 +/- 1%,and 93 +/- 4% of maximal contractile response at 5 min, for renal, mesenteric, and umbilical arteries, respectively). The AT1 receptor antagonist, losartan (10(-6) M), abolished contractile responses in renal and mesenteric arteries but only slowed the contraction in umbilical artery in response to ANG II and had no effect on maximal tension. AT2 receptor blockade (PD 123319, 10(-7) M) had no significant effect on the response to ANG II in any vessel. Indomethacin (10(-6) M) significantly potentiated contraction to ANG II in renal and mesenteric but not umbilical arteries. Northern and Western blot analyses demonstrated the presence of AT1 mRNA and protein in all three vessels. Immunostaining for the AT1 receptor was present in endothelium and the tunica media. These findings demonstrate the AT1 receptor is present and functionally active in fetal systemic arteries and are consistent with previous findings that the umbilical circulation displays a greater responsiveness to ANG II than the systemic vasculature.


Asunto(s)
Angiotensina II/farmacología , Feto/efectos de los fármacos , Arterias Umbilicales/efectos de los fármacos , Vasoconstricción/efectos de los fármacos , Angiotensina II/fisiología , Animales , Femenino , Feto/fisiología , Técnicas In Vitro , Arterias Mesentéricas/efectos de los fármacos , Arterias Mesentéricas/fisiología , Embarazo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptor de Angiotensina Tipo 1 , Receptor de Angiotensina Tipo 2 , Receptores de Angiotensina/genética , Receptores de Angiotensina/metabolismo , Arteria Renal/efectos de los fármacos , Arteria Renal/fisiología , Ovinos , Arterias Umbilicales/fisiología , Vasoconstricción/fisiología
10.
Gene ; 246(1-2): 209-18, 2000 Apr 04.
Artículo en Inglés | MEDLINE | ID: mdl-10767542

RESUMEN

The sequence from a human EST (IMAGE:259322) with homology to the nucleotide-sensitive chloride conductance regulator (ICln) was used to screen a human aortic cDNA library. The probe sequence was from a region of the EST lacking homology to ICln, and the goal was to isolate an ICln-like gene. A 2843bp cDNA clone with an open reading frame coding for a 561 amino acid protein was isolated. This clone had no homology to ICln. PROSITE analysis of the putative protein sequence reveals one tudor and two K homology (KH) domains. The gene has therefore been named TDRKH. Both KH and tudor motifs are involved in binding to RNA or single-strand DNA. PCR analysis demonstrated that TDRKH is alternatively spliced in several ways and alternatively polyadenylated at multiple sites. Northern analysis confirmed the presence of messages of multiple lengths with predominant bands at 2.8 and 4.0 kb and also demonstrated that TDRKH is widely expressed in human tissues. Within an intron of TDRKH, there is a region with 90% homology to ICln. This sequence, which is incorporated into the alternatively spliced message represented by IMAGE:259322, contains a 2 bp deletion that disrupts the ICln reading frame and therefore represents an ICln pseudogene. The TDRKH gene was mapped to the Epidermal Differentiation Complex (EDC) at chromosome 1q21 by radiation hybrid mapping and STS content of genomic clones from that region. The EDC contains a large cluster of related genes involved in terminal differentiation of the epidermis. It remains to be determined whether TDRKH has a specific role in epithelial function.


Asunto(s)
Empalme Alternativo , Canales Iónicos , Proteínas de Unión al ARN/genética , Secuencia de Aminoácidos , Secuencia de Bases , Sitios de Unión , Northern Blotting , Canales de Cloruro/genética , Mapeo Cromosómico , Cromosomas Humanos Par 1/genética , ADN/química , ADN/genética , ADN Complementario/química , ADN Complementario/genética , ADN Complementario/aislamiento & purificación , Feto/metabolismo , Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Genes/genética , Humanos , Intrones , Datos de Secuencia Molecular , Isoformas de Proteínas/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico , Distribución Tisular
11.
Am J Physiol Heart Circ Physiol ; 278(5): H1527-36, 2000 May.
Artículo en Inglés | MEDLINE | ID: mdl-10775130

RESUMEN

Anion currents contribute to vascular smooth muscle (VSM) membrane potential. The substitution of extracellular chloride (Cl) with iodide (I) or bromide (Br) initially inhibited and then potentiated isometric contractile responses of rat aortic rings to norepinephrine. Anion substitution alone produced a small relaxation, which occurred despite a lack of active tone and minimal subsequent contraction of endothelium-intact rings (4.2 +/- 1.2% of the response to 90 mM KCl). Endothelium-denuded rings underwent a similar initial relaxation but then contracted vigorously (I > Br). Responses to 130 mM I (93.7 +/- 1.9% of 90 mM KCl) were inhibited by nifedipine (10(-6) M), niflumic acid (10(-5) M), tamoxifen (10(-5) M), DIDS (10(-4) M), and HCO(-)(3)-free buffer (HEPES 10 mM) but not by bumetanide (10(-5) M). Intact rings treated with N(omega)-nitro-L-arginine (10(-4) M) responded weakly to I (15.5 +/- 2.1% of 90 mM KCl), whereas hemoglobin (10(-5) M), indomethacin (10(-6) M), 17-octadecynoic acid (10(-5) M), and 1H-[1,2, 4]oxadiazole[4,3-a]quinoxalin-1-one (10(-6) M) all failed to augment the response of intact rings to I. We hypothesize that VSM takes up I primarily via an anion exchanger. Subsequent I efflux through anion channels having a selectivity of I > Br > Cl produces depolarization. In endothelium-denuded or agonist-stimulated vessels, this current is sufficient to activate voltage-dependent calcium channels and cause contraction. Neither nitric oxide nor prostaglandins are the primary endothelial modulator of these anion channels. If they are regulated by an endothelium-dependent hyperpolarizing factor it is not a cytochrome P-450 metabolite.


Asunto(s)
Aorta/metabolismo , Endotelio Vascular/metabolismo , Canales Iónicos/metabolismo , Yoduro de Sodio/farmacología , Vasoconstricción/fisiología , Animales , Aorta/efectos de los fármacos , Bromuros/farmacología , Bloqueadores de los Canales de Calcio/farmacología , Canales de Cloruro/antagonistas & inhibidores , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/farmacología , Técnicas In Vitro , Canales Iónicos/efectos de los fármacos , Transporte Iónico/efectos de los fármacos , Masculino , Nifedipino/farmacología , Norepinefrina/farmacología , Cloruro de Potasio/farmacología , Ratas , Ratas Sprague-Dawley , Cloruro de Sodio/farmacología , Compuestos de Sodio/farmacología , Ésteres del Ácido Sulfúrico/farmacología , Vasoconstricción/efectos de los fármacos
12.
Magy Seb ; 53(6): 253-8, 2000 Dec.
Artículo en Húngaro | MEDLINE | ID: mdl-11299490

RESUMEN

During ten years 580 patients have been treated for gastric tumour in our department, 510 of them were operated on. Resection could be performed in 296 cases. 17 resections, 5.7 per cent of all were performed because of primary non-Hodgkin gastric lymphoma. No gastric lymphoma was found among the non-resected patients. The preoperative histological diagnosis was correct only in 8 cases. MALT origin could be proved in 5 patients. Synchronous adenocarcinoma and lymphoma was diagnosed in 2 patients. Staging was decided according to Lugano classification. There were six stage I, four stage II, and seven stage IV patients. 8 subtotal and 9 total gastrectomy was performed, 5 were extended and 2 were combined. R0 resection could be carried out in five stage I, two stage II and in one stage IV patient. We lost 2 patients in the postoperative period. Patients were treated with adjuvant chemotherapy (VEP, CHOP) except for 2 patients with low grade MALT lymphoma. The likelihood of one-year survival is 73 per cent, average two-year survival is 63 per cent. When the tumour is operable by total gastrectomy we suggest to perform splenectomy as well, despite of the fact that some postoperative complications can be related to it. We think it is reasonable to perform palliative resection in cases of locally extended stage IV tumours, which affect the patient's quality of life: to cease the pain, passage troubles, bleeding and to improve the conditions for adjuvant treatment.


Asunto(s)
Linfoma no Hodgkin/cirugía , Cuidados Paliativos/métodos , Neoplasias Gástricas/cirugía , Anciano , Anciano de 80 o más Años , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Quimioterapia Adyuvante , Femenino , Gastrectomía/efectos adversos , Humanos , Linfoma no Hodgkin/tratamiento farmacológico , Linfoma no Hodgkin/patología , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Estudios Retrospectivos , Neoplasias Gástricas/tratamiento farmacológico , Neoplasias Gástricas/patología , Análisis de Supervivencia , Resultado del Tratamiento
13.
J Mol Cell Cardiol ; 31(3): 657-66, 1999 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-10198195

RESUMEN

Chloride (Cl) ion channels play a critical role in the response of both vascular smooth muscle (VSM) and endothelial (ENDO) cells to agonist stimulation. In VSM, agonist-induced Cl currents produce membrane depolarization, resulting in calcium influx through voltage-sensitive channels. ENDO cells also activate Cl currents after either agonist application or perturbation of cell volume. Although some of these currents have been characterized biophysically, the genes involved have not been identified. The CLCN family of voltage-dependent Cl channel genes comprises nine members (CLCN1-7, Ka and Kb) which demonstrate quite diverse functional characteristics while sharing significant sequence homology. We used Northern-blot analysis to study the expression of these Cl channel genes in cultured human aortic and coronary VSM cells and in aortic ENDO cells. CLCN3 is by far the most abundant CLC channel mRNA in both VSM and ENDO cells. Lower levels of expression are seen for CLCN2, CLCN4, CLCN5 and CLCN6. Expression levels were similar in VSM and ENDO cells except for CLCN4 which was more highly expressed in ENDO cells. In situ hybridization was used to confirm the expression of CLCN3 in intact human fetal lung. CLCN3 message was seen in VSM and ENDO cells of both large and small pulmonary vessels, indicating that their detection by Northern blotting was not an artifact of cell culture. CLCN3 is also expressed in pulmonary epithelial and bronchial smooth muscle cells but not in chondrocytes or pulmonary interstitial cells. Recent studies suggest that CLCN3 may encode the swelling-induced Cl conductance. We used whole cell patch clamp recording to demonstrate swelling-induced Cl currents in these cultured VSM cells. This suggests that the CLCN3 protein is expressed; however, the functional role of this current in VSM remains to be determined.


Asunto(s)
Vasos Sanguíneos/metabolismo , Canales de Cloruro/genética , Northern Blotting , Canales de Cloruro CLC-2 , Diagnóstico por Imagen , Humanos , Hibridación in Situ , Pulmón/anatomía & histología , Pulmón/embriología , Pulmón/metabolismo , Familia de Multigenes/genética , Sondas de Oligonucleótidos , Técnicas de Placa-Clamp
14.
Am J Physiol ; 275(1): H151-60, 1998 07.
Artículo en Inglés | MEDLINE | ID: mdl-9688908

RESUMEN

Norepinephrine (NE) increases Cl- efflux from vascular smooth muscle (VSM) cells. An increase in Cl- conductance produces membrane depolarization. We hypothesized that if Cl- currents are important for agonist-induced depolarization, then interfering with cellular Cl- handling should alter contractility. Isometric contraction of rat aortic rings was studied in a bicarbonate buffer. Substitution of extracellular Cl- with 130 mM methanesulfonate (MS; 8 mM Cl-) did not cause contraction. NE- and serotonin-induced contractions were potentiated in this low-Cl- buffer, whereas responses to K+, BAY K 8644, or NE in the absence of Ca2+ were unaltered. Substitution of Cl- with I- or Br- suppressed responses to NE. Inhibition of Cl- transport with bumetanide (10(-5) M) or bicarbonate-free conditions (10 mM HEPES) inhibited NE- but not KCl-induced contraction. The Cl--channel blockers DIDS (10(-3) M), anthracene-9-carboxylic acid (10(-3) M), and niflumic acid (10(-5) M) all inhibited NE-induced contraction, whereas tamoxifen (10(-5) M) did not. Finally, disruption of sarcoplasmic reticular function with cyclopiazonic acid (10(-7) M) or ryanodine (10(-5) M) prevented the increase in the peak response to NE produced by low-Cl- buffer. We conclude that a Cl- current with a permeability sequence of I- > Br- > Cl- > MS is critical to agonist-induced contraction of VSM.


Asunto(s)
Aorta Torácica/fisiología , Canales de Cloruro/fisiología , Cloruros/farmacología , Contracción Isométrica/fisiología , Músculo Liso Vascular/fisiología , Norepinefrina/farmacología , Vasoconstricción/fisiología , Ácido 3-piridinacarboxílico, 1,4-dihidro-2,6-dimetil-5-nitro-4-(2-(trifluorometil)fenil)-, Éster Metílico/farmacología , Ácido 4,4'-Diisotiocianostilbeno-2,2'-Disulfónico/farmacología , Animales , Aniones/metabolismo , Antracenos/farmacología , Aorta Torácica/efectos de los fármacos , Bicarbonatos/farmacología , Permeabilidad de la Membrana Celular , Canales de Cloruro/antagonistas & inhibidores , Técnicas In Vitro , Indoles/farmacología , Contracción Isométrica/efectos de los fármacos , Masculino , Mesilatos/farmacología , Músculo Liso Vascular/efectos de los fármacos , Ácido Niflúmico/farmacología , Potasio/farmacología , Ratas , Ratas Sprague-Dawley , Rianodina/farmacología , Retículo Sarcoplasmático/efectos de los fármacos , Retículo Sarcoplasmático/fisiología , Serotonina/farmacología , Vasoconstricción/efectos de los fármacos , Vasodilatadores/farmacología
15.
Am J Physiol ; 275(1): H161-8, 1998 07.
Artículo en Inglés | MEDLINE | ID: mdl-9688909

RESUMEN

Activation of a Cl- current is critical to agonist-induced activation of rat aortic smooth muscle contraction. Substituting extracellular Cl- with 130 mM methanesulfonate (8 mM Cl-) increases the contractile response to norepinephrine (NE) but not to KCl. We hypothesized that endothelial factors modulate this effect. Removing the endothelium (rubbing) or treatment with N-nitro L-arginine (L-NNA) markedly increased the potentiation of NE-induced contraction by low-Cl- buffer. Indomethacin had no effect. The previously demonstrated ability of Cl--channel blockers (DIDS, anthracene-9-carboxylic acid, niflumic acid) or Cl- transport inhibitors (bumetanide, bicarbonate-free buffer) to inhibit responses to NE was not altered by L-NNA. Low-Cl- buffer alone did not contract intact rings but produced nifedipine-sensitive contractile responses after rubbing or L-NNA treatment. These data suggest that the Cl- conductance of smooth muscle in intact blood vessels is low but increases with withdrawal of reduced nitric oxide (NO') or agonist stimulation. Rubbing or L-NNA increased the sensitivity of rings to KCl but not to NE. Nifedipine reduced both sensitivity and maximum response to NE in intact vessels. L-NNA increased the maximum response to NE in nifedipine-treated rings without changing sensitivity. We conclude that although NO' affects both the voltage-dependent and voltage-independent components of contraction, sensitivity to NE is determined by the voltage-dependent portion. The voltage change required for a full response to NE is dependent on activation of a Cl- current that may be under the tonic regulatory influence of NO'.


Asunto(s)
Aorta Torácica/fisiología , Canales de Cloruro/fisiología , Endotelio Vascular/fisiología , Contracción Isométrica/fisiología , Músculo Liso Vascular/fisiología , Norepinefrina/farmacología , Ácido 4,4'-Diisotiocianostilbeno-2,2'-Disulfónico/farmacología , Animales , Antracenos/farmacología , Aorta Torácica/efectos de los fármacos , Canales de Cloruro/antagonistas & inhibidores , Técnicas In Vitro , Indometacina/farmacología , Contracción Isométrica/efectos de los fármacos , Masculino , Modelos Cardiovasculares , Músculo Liso Vascular/efectos de los fármacos , Nifedipino/farmacología , Ácido Niflúmico/farmacología , Óxido Nítrico/fisiología , Nitroarginina/farmacología , Ratas , Ratas Sprague-Dawley , Vasoconstricción/efectos de los fármacos
16.
Orv Hetil ; 139(51): 3079-83, 1998 Dec 20.
Artículo en Húngaro | MEDLINE | ID: mdl-9914729

RESUMEN

GIP is a rarely occurring disorder. There is only few literature from its first description. Authors observed the course of GIP in the case of a 54 year old female patient in the form of bilateral disseminated microfocal pulmonary shadows, increased reticular outline with associated respiratory insufficiency. Open fine needle pulmonary biopsy proved giant cell desquamative alveolitis with help of light- and electronmicroscopical and histochemical examinations. Although possibility of exogenic, inhalative factor or/and infectious origin arose in causing the disease, disposition from the patient's actual immunological status could had helped the evolution of the disease. This fact seemed to be supported by the histologically proven associated dermatitis purpurica pigmentosa (Schamberg disease). With methylprednisolon therapy full radiological recovery occurred, while Schamberg disease was little influenced by the above mentioned therapy. The patient is pulmonologically symptom-free and without complaint after 1 year without any steroid-medication.


Asunto(s)
Neumonía en Organización Criptogénica/complicaciones , Enfermedades Pulmonares Intersticiales , Fibrosis Pulmonar/inmunología , Neumonía en Organización Criptogénica/diagnóstico por imagen , Neumonía en Organización Criptogénica/tratamiento farmacológico , Neumonía en Organización Criptogénica/inmunología , Femenino , Células Gigantes , Humanos , Enfermedades Pulmonares Intersticiales/complicaciones , Enfermedades Pulmonares Intersticiales/diagnóstico por imagen , Enfermedades Pulmonares Intersticiales/tratamiento farmacológico , Enfermedades Pulmonares Intersticiales/inmunología , Metilprednisolona/uso terapéutico , Microscopía Electrónica , Persona de Mediana Edad , Alveolos Pulmonares/patología , Fibrosis Pulmonar/diagnóstico por imagen , Fibrosis Pulmonar/tratamiento farmacológico , Radiografía Torácica , Resultado del Tratamiento
17.
Biochemistry ; 36(49): 15489-500, 1997 Dec 09.
Artículo en Inglés | MEDLINE | ID: mdl-9398278

RESUMEN

A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA was screened for galactanase-positive recombinants. The nine galactanase positive phage isolated contained the same galactanase gene designated galA. The deduced primary structure of the enzyme (galactanase A; GalA) encoded by galA had a Mr of 42 130 and exhibited significant sequence identity with a galactanase from Aspergillus aculeatus, placing GalA in glycosyl hydrolase family 53. The enzyme displayed properties typical of an endo-beta1, 4-galactanase and exhibited no activity against the other plant structural polysaccharides evaluated. Analysis of the stereochemical course of 2,4-dinitrophenyl-beta-galactobioside (2,4-DNPG2) hydrolysis by GalA indicated that the galactanase catalyzes the hydrolysis of glycosidic bonds by a double displacement general acid-base mechanism. Hydrophobic cluster analysis (HCA) suggested that family 53 enzymes are related to the GH-A clan of glycosyl hydrolases, which have an (alpha/beta)8 barrel structure. HCA also predicted that E161 and E270 were the acid-base and nucleophilic residues, respectively. Mutants of GalA in which E161 and E270 had been replaced with alanine residues were essentially inactive against galactan. Against 2,4-DNPG2, E161A exhibited a much lower Km and kcat than native GalA, while E270A was inactive against the substrate. Analysis of the pre-steady-state kinetics of 2,4-DNPG2 hydrolysis by E161A showed that there was an initial rapid release of 2,4-dinitrophenol (2,4-DNP), which then decayed to a slow steady-state rate of product formation. No pre-steady-state burst of 2,4-DNP release was observed with the wild-type enzyme. These data are consistent with the HCA prediction that E161 and E270 are the acid-base and nucleophilic catalytic residues of GalA, respectively.


Asunto(s)
Glicósido Hidrolasas/metabolismo , Pseudomonas fluorescens/enzimología , beta-Galactosidasa/metabolismo , Secuencia de Aminoácidos , Proteínas Bacterianas , Secuencia de Bases , Catálisis , ADN Recombinante , Galactanos/metabolismo , Regulación Bacteriana de la Expresión Génica , Regulación Enzimológica de la Expresión Génica , Glicósido Hidrolasas/química , Hidrólisis , Espectroscopía de Resonancia Magnética , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Homología de Secuencia de Aminoácido , beta-Galactosidasa/química
18.
Orv Hetil ; 137(10): 507-11, 1996 Mar 10.
Artículo en Húngaro | MEDLINE | ID: mdl-8713664

RESUMEN

Authors investigated the presence of antibody to hepatitis C virus (anti-HCV) at 67,534 blood donations, and alanin-amino-transferase (ALT) levels were analysed at the same time. The prevalence of anti-HCV antibody was 0.73% at 15,864 blood donors. Frequency of positive reactions of anti-HCV antibody increased paralelly with advance of age. Increased or high serum ALT level was found at more than 50% of samples with positive HCV reactivity. At the base of follow-up the blood donors were screened out from blood donation temporarily or definitively. Anti-HCV donors were taken to hepatic care. The treatment with interferon of HCV seropositive donors proved by PCR (polimerase chain reaction) was started. Transfusion and tattou are underlined as risk factors of HCV infection. The data were analysed according to sex and ages.


Asunto(s)
Donantes de Sangre , Transfusión Sanguínea , Hepacivirus/inmunología , Anticuerpos contra la Hepatitis C/inmunología , Adolescente , Adulto , Anciano , Alanina Transaminasa/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Hepatitis C/inmunología , Hepatitis C/transmisión , Humanos , Hungría , Masculino , Persona de Mediana Edad , Prevalencia , Factores de Riesgo
19.
Orv Hetil ; 137(1): 9-14, 1996 Jan 07.
Artículo en Húngaro | MEDLINE | ID: mdl-8721857

RESUMEN

Authors analyse complaints of 166 patients suffering in gastroesophageal reflux disease, and results of different objective diagnostic methods (X-ray swallowing, endoscopy, 24 hour pH-monitoring, esophageal manometry). The degree of severity of typical complaints and changes found by diagnostic procedures were scored. Individual diagnostic proceduresindicated the characteristic changes of gastroesophageal reflux with great deviations. Hiatal hernia identified by x-ray, esophagitis observed by endoscopy, abnormal reflux revealed by pH-metry together occurred only at 36% of patients. Significant correlation could be revealed only between the degree of changes found by x-ray or pH-metry--applying them one by one--and the complaints of the patients, but with the combination of four diagnostic methods a tight, positive correlation could be demonstrated with the severity of the complaints of the patients in every case. Applying of diagnostic methods as much as possible completing each others, with consideration to the complaints of the patients is suggested for having the diagnosis been precise of the gastroesophageal reflux disease, and for measuring of efficacy of treatment. The importance of tight collaboration of internist, radiologist and surgeon and the significance of complex examination is underlined.


Asunto(s)
Reflujo Gastroesofágico/diagnóstico , Esofagoscopía , Reflujo Gastroesofágico/fisiopatología , Humanos , Concentración de Iones de Hidrógeno , Manometría , Monitoreo Fisiológico
20.
Anesthesiology ; 76(2): 236-44, 1992 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-1346560

RESUMEN

Neuromuscular blockers may affect cerebral blood flow (CBF) regulation in the newborn. We studied the effects of d-tubocurarine (0.1 mg.kg-1, n = 8), pancuronium (0.1 and 0.4 mg.kg-1, n = 6 and 7), and vecuronium (0.1 and 0.4 mg.kg-1, n = 6 and 7) on CBF measured over the same range of mean systemic blood pressure ([BP] 15-122 mmHg) in each group of newborn pigs; controls received normal saline (n = 7). The levels of BP during hypotension and hypertension were scaled at intervals of 5 +/- 1.6 mmHg and adjusted by inflating balloon-tipped catheters placed in the aorta. After saline, the low dose of pancuronium (0.1 mg.kg-1), and the two doses of vecuronium, CBF was constant over the BP range of 50-90 mmHg (r = -0.07-0.35, P greater than 0.20) but varied directly with BP beyond this range (tau = 0.38 - 0.60, P less than 0.05). In contrast, in pigs treated with d-tubocurarine and high-dose pancuronium, CBF remained constant from 35 to 122 mmHg of BP (r = 0.14 - 0.37, P greater than 0.10) and changed minimally (4-12%) with BP greater than 105 mmHg compared to the other groups (41-59%, P less than 0.01). When BP was reduced below 30 mmHg, CBF also decreased less (20-38%) in animals treated with d-tubocurarine and high dose-pancuronium than after the other treatments (58-67%, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Asunto(s)
Animales Recién Nacidos/fisiología , Circulación Cerebrovascular/fisiología , Homeostasis/efectos de los fármacos , Pancuronio/farmacología , Tubocurarina/farmacología , Bromuro de Vecuronio/farmacología , Animales , Presión Sanguínea/fisiología , Circulación Cerebrovascular/efectos de los fármacos , Estimulación Química
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