RESUMEN
The aim of this study was to describe the effect of unilateral orchidectomy on testicular characteristics of mature domestic cats. Five, 1- to 2-year-old, cross-bred male cats were unilaterally orchidectomized on day 0 (right) and day 60 (left). The testes were processed for histological and immunohistochemical evaluation and groups compared by Student's t-test. No significant differences (p > 0.05) were found between testes groups for any of the gross (volume and gonadosomatic index) and microscopic (tubular diameters, axis, area, length and perimeter, germinal epithelium height, spermatogonias, spermatocytes, spermatids, spermatozoa, Sertoli and Leydig cells, intertubular compartment, lumen, cellular debris, tubular- intertubular proportion and basement membrane) parameters assessed. According to these biometric and morphometric results, mature cats do not develop compensatory hypertrophy within 60 days of unilateral orchidectomy.
Asunto(s)
Gatos/fisiología , Hipertrofia/veterinaria , Orquiectomía/veterinaria , Testículo/fisiología , Envejecimiento , Animales , Masculino , Orquiectomía/métodosRESUMEN
La Enfermedad de Chagas constituye un problema de salud pública en Venezuela, cuyas alteraciones están representadas fundamentalmente por el desarrollo de miocardiopatías incapacitantes. En la actualidad, las causas de las miocardiopatías no son realmente conocidasy tampoco existe tratamiento; sin embargo, dentro de la teoria neurogénica se ha postulado que el Sistema Colinérgico est involúcrado en su desarrollo. Con el fin de dilucidar el estado de Receptor Colinérgico Muscarínico (RCM) en ésta enfermedad, se establecieron dos grupos de animales: Control (ratas sanas) y Experimental (ratas con Enfermedad de Chagas en etapa crónica), posteriormente las ratas fueron sacrificadas, disecando en corazón Ventrículo Izquierdo (VI), Ventrículo Derecho (VD), Tabique Interventricular (TIV), Aurícula (Au) y en el encéfalo Tronco Encefálico (TE). Se determinó la cantidad de proteínas mediante el método de Lowry, la densidad de RCM y los efectos de agonistas (Carbacol) y antagonistas selectivos (Pirenzepina, Metoctramina, 4-DAMP y Tropicamida) por medio de ensayos de radioligandos usando [³H]-QNB como marcador radioactivo. Los resultados mostraron en TE una disminución significativa de la cantidad de proteínas en ratas con Enfermedad de Chagas; en VD se encontró una supersensibilidad significativa del RCM en ratas Chagásicas, mientras que en TIV, VI y Au no se observaron diferencias significativas entre ambos grupos. En las curvas de desplazamiento de [³H]-QNB por agonistas y antagonistas selectivos, no se encontraron diferencias significativas en el perfil de desplazamiento y en los valores de CI50, al comparar los dos grupos. En conclusión la expresión de proteínas y del RCM se encuentran alteradas en TE y VD, respectivamente.
Asunto(s)
Animales , Ratas , Cardiomiopatías , Enfermedad de Chagas , Ratas Sprague-Dawley , Salud Pública , VenezuelaRESUMEN
Angiotensin II (Ang II)-induced phospholipase D (PLD) activity is greater in aortic smooth muscle from spontaneously hypertensive rats (SHR) versus normotensive Wistar-Kyoto rats (WKY). Whether and how this signaling pathway is altered in preglomerular microvascular smooth muscle cells (PGSMCs), a cell type that may participate in genetic hypertension, is unknown. The goals of the present study were to determine in SHR and WKY PGSMCs the following: (1) whether Ang II induces PLD activity; (2) whether the effect of Ang II on PLD activity is greater in SHR; (3) which PLD isoform is stimulated by Ang II; (4) what signaling pathway mediates Ang II-induced PLD stimulation; and (5) whether the signaling pathways mediating Ang II-induced PLD activity are different in SHR and WKY. The EC(50) for Ang II-induced PLD stimulation in SHR was 10-fold lower than the EC(50) in WKY, and both were inhibited by L-158,805, an AT(1) antagonist. Inhibitors of phosphoinositol-3-kinase and protein kinase C did not block Ang II-induced PLD activity in SHR and WKY PGSMCs. Catalytically-inactive constructs of PLD2 and RhoA, but not PLD1, ADP ribosylation factor 1 (ARF1), ARF6, or ADP ribosylation factor nucleotide exchange factor (ARNO) blocked Ang II-induced PLD activity in SHR and WKY PGSMCs. Brefeldin A completely blocked Ang II-induced PLD activity in SHR but only slightly reduced Ang II-induced PLD activity in WKY PGSMCs. Therefore, we conclude that in PGSMCs, the effect of Ang II on PLD activity is (1) greater in SHR; (2) mediated by AT(1) receptors signaling to PLD2; (3) transduced primarily by Rho proteins; and (4) inhibited in SHR by brefeldin A.
Asunto(s)
Angiotensina II/farmacología , Músculo Liso Vascular/efectos de los fármacos , Fosfolipasa D/biosíntesis , Arteria Renal/efectos de los fármacos , Angiotensina I/antagonistas & inhibidores , Antagonistas de Receptores de Angiotensina , Animales , Brefeldino A/farmacología , Células Cultivadas , Hipertensión/fisiopatología , Modelos Químicos , Músculo Liso Vascular/enzimología , Fosfolipasa D/antagonistas & inhibidores , Inhibidores de la Síntesis de la Proteína/farmacología , ARN Mensajero/biosíntesis , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Receptor de Angiotensina Tipo 1 , Receptor de Angiotensina Tipo 2 , Arteria Renal/enzimología , Transducción de SeñalRESUMEN
STATEMENT OF THE PROBLEM: Numerous articles emphasize the importance of passivity of implant-prosthetic component interfaces. Nonpassive interfaces can lead to bone loss, abutment fracture, and connecting screw breakage. PURPOSE: The purpose of this study was to evaluate 3 postcasting techniques for the correction of non-passive fit between a cast bar superstructure and its interface with an implant abutment. MATERIAL AND METHODS: Thirty implant Hader bars were fabricated based on a metal model composed of two 3.8/4.5 HL PME titanium implant abutments. Initial measurements were collected on the y-axis of the left implant abutment-bar interface by using a M2001ARS toolmaker microscope. Means were calculated from buccal, distal, and lingual measurements on each specimen. Ten specimens were sectioned, indexed, and corrected by casting the same alloy (group 1). Ten specimens were sectioned, indexed, and corrected by soldering (group 2). The last 10 specimens were submitted to 2 cycles of electrical discharge machining on a MedArc M-2 EDM machine (group 3). Postcorrection measurements were collected on the 3 groups. A 1-way ANOVA and a Tukey-Kramer test at a 0.05 significance level were performed on the 3 groups after the corrective techniques. RESULTS: Initial gap means were 192 microm for group 1, 190 microm for group 2, and 198 microm for group 3. There was a significant difference (P<0.05) in gap means between group 1 (15 microm) and group 2 (72 microm) as well as between group 2 and group 3 (7.5 microm) after each correction technique. No difference was detected between group 1 and group 3. CONCLUSION: The electrical discharge machining group resulted in the smallest mean gap distance of 7.5 microm, thus meeting the criteria of passive fit (within 10 microm) described in the literature.
Asunto(s)
Técnica de Colado Dental , Prótesis Dental de Soporte Implantado/instrumentación , Análisis de Varianza , Pilares Dentales/estadística & datos numéricos , Técnica de Colado Dental/normas , Técnica de Colado Dental/estadística & datos numéricos , Técnica de Impresión Dental , Prótesis Dental de Soporte Implantado/métodos , Prótesis Dental de Soporte Implantado/normas , Prótesis Dental de Soporte Implantado/estadística & datos numéricos , Soldadura Dental/métodos , Diseño de Dentadura/métodos , Diseño de Dentadura/normas , Diseño de Dentadura/estadística & datos numéricos , Dentadura Parcial Removible/normas , Dentadura Parcial Removible/estadística & datos numéricos , Aleaciones de Oro , Humanos , TitanioRESUMEN
This study tests the hypothesis that activation of protein kinase C (PKC) is a critical step for early recovery from spontaneous nystagmus after unilateral ablation of the vestibular periphery. Halothane-NO(2)-O(2)-anesthetized Long-Evans rats received a 5-microl intracerebroventricular bolus of vehicle (distilled water, six rats), PKC inhibitor [Iso-H-7 (10 mM, four rats; 50 mM, five rats) or bisindolemaleimide I (Bis-I, 10 microM six rats)], PKG and PKA inhibitor (A-3, 1 mM, six rats), or the serine-threonine protein kinase inhibitor H-7 (1 mM, five rats; 10 mM, five rats). Surgical unilateral labyrinthectomy (UL) was completed within 15 min. Sham control groups showed no nystagmus. Bis-I and Iso-H-7 significantly retarded the disappearance of spontaneous nystagmus quick phases for 8 h after UL (p<0.05). The effects of Iso-H-7 were dose-dependent: more nystagmus quick phases (p<0.05) were present in the 50 mM than the 10 mM group at 7 and 8 h post-UL. The rats given A-3 showed a delayed retardation of nystagmus loss, which differed significantly (p<0.05) from controls at 4-8 h after labyrinthectomy. The number of nystagmus quick phases was significantly greater than controls (p<0. 05) in the 10 mM H-7 group at 4, 5, 6 and 48 h post-UL, but only at 6 and 24 h post-UL in the 1 mM H-7 group. Thus, PKC activation is an important early requirement for vestibular compensation during the acute post-labyrinthectomy period, while cyclic-nucleotide dependent kinases may be important in a later time frame.
Asunto(s)
Oído Interno/fisiología , Nistagmo Patológico/metabolismo , Proteína Quinasa C/antagonistas & inhibidores , Células de Purkinje/enzimología , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/análogos & derivados , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/farmacología , Animales , Oído Interno/cirugía , Inhibidores Enzimáticos/farmacología , Indoles/farmacología , Inyecciones Intraventriculares , Masculino , Maleimidas/farmacología , Células de Purkinje/efectos de los fármacos , Ratas , Ratas Long-Evans , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiologíaRESUMEN
The behavioral recovery from unilateral labyrinthectomy (UL) in rats is accompanied by asymmetric expression of Protein kinase C (PKC) in parasagittal regions of the flocculonodular lobe within 6 h after UL, which resolves to the control, symmetric pattern within 24 h. These changes consist of a regionally selective increase in the number of PKC-immunopositive Purkinje cells contralateral to the lesion. This study tested the hypotheses (1) that climbing fiber innervation inhibits PKC expression and (2) that climbing fibers are essential for the observed changes in PKC expression within 6 h after UL. The patterns of flocculonodular lobe Purkinje cell PKCdelta expression were analyzed 6 h post-operatively in both UL and sham-operated that had been treated previously with 3-acetylpyridine to destroy the inferior olive. These data were compared with previous results from rats with an intact olive. The results suggest that at least two signals regulate the zonal distribution of Purkinje cell PKCdelta expression in the flocculonodular lobe during the early period of compensation from UL. Climbing fiber activation appears to reduce PKC expression, while extraolivary mechanisms appear to up-regulate PKC expression. It is suggested that the climbing fiber signals may act as a molecular 'filter' or 'automatic gain control' which adjusts the contributions of these kinases to synaptic plasticity within the context of the background activity of climbing fibers.
Asunto(s)
Núcleos Cerebelosos/enzimología , Núcleos Cerebelosos/fisiología , Fibras Nerviosas/fisiología , Proteína Quinasa C/biosíntesis , Vestíbulo del Laberinto/fisiología , Animales , Núcleos Cerebelosos/citología , Oído Interno/fisiología , Procesamiento de Imagen Asistido por Computador , Inmunohistoquímica , Masculino , Vías Nerviosas/citología , Vías Nerviosas/fisiología , Células de Purkinje/fisiología , RatasRESUMEN
The objectives of the present study were to determine whether angiotensin II (Ang II) modifies beta-adrenoceptor-induced cAMP production in preglomerular microvascular smooth muscle cells (PMVSMCs), to determine whether the Ang II/beta-adrenoceptor interaction on cAMP production differs in PMVSMCs from normotensive Wistar-Kyoto (WKY) rats vs. PMVSMCs from spontaneously hypertensive rats (SHR), and to elucidate the mechanism of Ang II/beta-adrenoceptor interactions on cAMP production in PMVSMCs. In cultured PMVSMCs, isoproterenol increased cAMP levels and this effect was markedly enhanced by Ang II. The Ang II enhancement of isoproterenol-induced cAMP was significantly greater in SHR PMVSMCs compared with WKY PMVSMCs. Neither inhibition of calcineurin with FK506, inhibition of calcium-calmodulin with W-7 and calmidazolium, nor inhibition of Gi proteins with pertussis toxin attenuated Ang II enhancement of isoproterenol-induced cAMP in PMVSMCs from either SHR or WKY rats. Moreover, the effect of Ang II on isoproterenol-induced cAMP was not mimicked by alpha-2 adrenoceptor stimulation. In contrast, chelation of intracellular calcium with BAPTA-AM attenuated, increasing intracellular calcium with A23187 augmented, and inhibition of protein kinase C with either calphostin C or chelerythrine chloride abolished Ang II enhancement of isoproterenol-induced cAMP. We conclude that in cultured PMVSMCs Ang II enhances the cAMP response to beta-adrenoceptor agonists via a mechanism that involves coincident activation of adenylyl cyclase by stimulatory G proteins and protein kinase C. Thus, protein kinase C-mediated activation of adenylyl cyclase may attenuate Ang II-induced vasoconstriction in the renal microcirculation by raising the intracellular levels of cAMP, and this mechanism may be augmented in genetic hypertension.
Asunto(s)
Angiotensina II/farmacología , AMP Cíclico/biosíntesis , Hipertensión/metabolismo , Isoproterenol/farmacología , Glomérulos Renales/irrigación sanguínea , Músculo Liso Vascular/metabolismo , Animales , Calcio/metabolismo , Calmodulina/fisiología , Proteínas de Unión al GTP/fisiología , Masculino , Microcirculación/metabolismo , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKYRESUMEN
Protein kinase C (PKC) is a family of intracellular signal transduction enzymes, comprising isoforms that vary in sensitivity to calcium, arachidonic acid, and diacylglycerol. PKC isoforms alpha, gamma, and delta are expressed by cerebellar Purkinje cells and neurons in the cerebellar nuclei and vestibular nuclei of the Long-Evans rat. In control rats, these PKCs are distributed symmetrically in the flocculonodular-lobe Purkinje cells. Behavioral recovery from vestibular dysfunction produced by unilateral labyrinthectomy (UL) is accompanied by asymmetric expression of PKC isoforms in these regions within 6 hr after UL. These expression changes were localized within parasagittal regions of the flocculus and nodulus. The distribution of PKCalpha, -gamma, and -delta were identical, suggesting that they are coregulated in cerebellar Purkinje cells during this early compensatory period. The pattern of Purkinje cell PKC expression returned to the control, symmetric distribution within 24 hr after UL. It is hypothesized that these regional changes in Purkinje cell PKC expression are an early intracellular signal contributing to vestibular compensation. In particular, regulation of PKC expression may contribute to changes in the efficacy of cerebellar synaptic plasticity during the acute post-UL period.
Asunto(s)
Proteína Quinasa C/metabolismo , Células de Purkinje/enzimología , Vestíbulo del Laberinto/fisiología , Animales , Especificidad de Anticuerpos , Vías Auditivas/fisiología , Western Blotting , Núcleos Cerebelosos/citología , Núcleos Cerebelosos/enzimología , Oído Interno/fisiología , Oído Interno/cirugía , Masculino , Proteína Quinasa C/análisis , Proteína Quinasa C/inmunología , Ratas , Ratas Endogámicas , Transducción de Señal/fisiologíaRESUMEN
The mechanism by which lovastatin lowers cholesterol levels is well characterized but little is known about its anti-mitogenic and anti-tumorigenic mechanism. Here we demonstrate that lovastatin disrupts early events in the mitogenic signaling pathways of insulin. Insulin treatment (200 mM) of quiescent HIR rat-1 fibroblasts results in an 8-fold stimulation of phosphatidylinositol-3-kinase (PI-3-K) activity. Overnight pretreatment of cells with lovastatin (20 microM) inhibits insulin stimulation of PI-3-K activity by 75%. Immunoprecipitation and immunoblotting experiments using antibodies against the regulatory subunit of PI-3-K (p85), phosphotyrosine, and insulin receptor alpha and beta subunits demonstrate that lovastatin inhibits the association of p85 with tyrosine phosphorylated insulin receptor substrate-1 and the beta subunit of the insulin receptor. Furthermore, lovastatin dramatically reduces (70-100%) the level of tyrosine phosphorylated insulin receptor beta subunit following insulin stimulation. These results clearly demonstrate that lovastatin disrupts early events of insulin mitogenic signaling by reducing the levels of tyrosine phosphorylated beta subunit and suggest that this disruption is a potential mechanism for the anti-mitogenic effect of lovastatin.
Asunto(s)
Insulina/farmacología , Lovastatina/farmacología , Receptor de Insulina/fisiología , Transducción de Señal/efectos de los fármacos , Animales , Línea Celular , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Humanos , Proteínas Sustrato del Receptor de Insulina , Cinética , Sustancias Macromoleculares , Fosfatidilinositol 3-Quinasas , Fosfoproteínas/metabolismo , Fosfotransferasas (Aceptor de Grupo Alcohol)/metabolismo , Fosfotirosina , Ratas , Receptor de Insulina/biosíntesis , Transducción de Señal/fisiología , Transfección , Tirosina/análogos & derivados , Tirosina/metabolismoRESUMEN
Twenty-six isolates of Leishmania parasites of Peruvian origin were studied by isoenzyme electrophoresis of four marker enzymes (ASAT, ALAT, G6PD and GPI), kinetoplast DNA hybridization and monoclonal antibody binding and compared with marker strains of the New World organisms L. b. braziliensis, L. b. guyanensis, L. m. mexicana and L. m. amazonensis. 12 of the isolates studied were of Andean origin; 11 of these were isolated from patients with Andean cutaneous leishmaniasis. The organisms originating from the Peruvian Amazonian forest were isolated from patients with cutaneous (12 cases) or mucocutaneous (2 cases) leishmaniasis. One of the Andean isolates was obtained from an infected phlebotomine vector. 25 of the new isolates were identified as L. braziliensis ssp. according to the three techniques employed. The results of monoclonal antibody binding showed that 23 of the isolates were indistinguishable from L. b. braziliensis. Two isolates identified as L. braziliensis ssp. according to their isoenzyme profiles and k-DNA hybridization patterns could not be classified at the subspecies level. The isolate obtained from the phlebotomine vector could not be identified. No evidence of the existence of parasites of the L. mexicana complex in Peruvian territory was found in this study. The results obtained show a remarkable similarity between Leishmania of Andean origin and L. b. braziliensis.