RESUMEN
AIM: The aim of the present study was to perform a comparative analysis of occlusal contact points in children with and without signs and symptoms of Temporomandibular Disorders (TMD). STUDY DESIGN: Cross-sectional study. One hundred fifty children between 6 and 14 years of age were evaluated using the Helkimo questionnaire and a clinical exam. The occlusal contact points in each child were recorded during maximal intercuspation with the aid of carbon strips. Digital photographs were taken of the upper and lower arches before and after recording the occlusal contacts. The number of contact points between sides were compared and recorded on individual charts (occlusograms). STATISTICS: Student's t-test and Pearson's chi-square test were used for the statistical analysis, with the level of significance set at 0.05, which revealed no statistically significant differences between genders. The Student's t-test revealed a statistically significant difference in the mean number of occlusal contact points between the participants with and without TMD, with a higher number of contact points among those without TMD. There was no significant difference between sides. RESULTS The results of this study show a difference in the number of occlusal contact points in centric occlusion between children with and without TMD. CONCLUSION Regardless of the degree of severity, the number of occlusal contact points is lower among children with TMD.
Asunto(s)
Oclusión Dental Céntrica , Registro de la Relación Maxilomandibular , Trastornos de la Articulación Temporomandibular/patología , Adolescente , Niño , Estudios Transversales , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador/métodos , Registro de la Relación Maxilomandibular/instrumentación , Masculino , Músculos Masticadores/patología , Músculos del Cuello/patología , Palpación , Fotografía Dental/métodos , Proyectos Piloto , Rango del Movimiento Articular/fisiología , Articulación Temporomandibular/patología , Trastornos de la Articulación Temporomandibular/diagnósticoRESUMEN
The primary objective of this study was to compare expression of maternal transcripts in bovine oocyte populations with differential developmental competence: oocytes from prepubertal and pubertal animals; and oocytes from small (3-4 mm) and large (6-10 mm) follicles from pubertal animals. All transcripts were examined in oocytes prior to and after in vitro maturation (IVM). Genes were selected based on their known maternal effect in mouse (ZAR1, STELLA, HSF1, MATER/NLRP5 and its paralogue NLRP9), or their identification as markers of oocyte maturation, either involved in redox metabolism (PRDX1, PRDX2) or meiotic progression (AURKA). Total or polyadenylated forms of the transcripts were followed by reverse transcription coupled to real-time PCR. Six polyadenylated transcripts were found significantly reduced after maturation irrespective of donor age or follicle diameter (p<0.05). Within these six polyadenylated transcripts, ZAR1, NLRP9, HSF1, PRDX1 and PRDX2 were significantly reduced in oocytes from prepubertal animals compared to adult animals (p<0.05). A younger age was also associated with lower abundance (total form) of PRDX2/PRDX1 irrespective of maturation. Total HSF1, PRDX1 and polyadenylated NLRP9 showed a tendency (p values from 0.053 to 0.08) for a higher detection in oocytes from small follicles, thus encouraging further investigation of the follicle diameter model. However, at the present time, follicle size did not significantly affect expression of transcripts examined. In conclusion, this study demonstrates differences in the maternal store of RNA and its regulation during IVM which is dependent on donor age.
Asunto(s)
Bovinos , Perfilación de la Expresión Génica , Expresión Génica , Oocitos/metabolismo , ARN Mensajero/análisis , Maduración Sexual , Envejecimiento , Animales , Femenino , Meiosis/genética , Oocitos/crecimiento & desarrollo , Folículo Ovárico/anatomía & histología , Peroxidasas/genética , Peroxirredoxinas/análisis , Reacción en Cadena de la Polimerasa/veterinaria , Maduración Sexual/genéticaRESUMEN
Malassezia spp. may act as opportunistic skin pathogens in humans and animals. Malassezia pachydermatis proliferation and phospholipase production may play a pathogenic role in the occurrence of skin lesions in dogs. This study investigates the presence of mu-opioid receptor (MOR) in M. pachydermatis strains isolated from healthy dogs and dogs with skin lesions and its effects on phospholipase activity (p.a.). P.a. of 64 M. pachydermatis isolates was evaluated using different concentrations of naloxone (Nx), a MOR antagonist. Isolates were divided into Group A (i.e., 40 isolates from 26 dogs with dermatitis) and Group B (i.e., 24 isolates from 12 healthy dogs). The MOR expression was analyzed by Western blot and immunofluorescence. A statistically higher p.a. than that of the controls was found with isolates in Group A at a Nx concentration of 10(-6) M (P<0.05). No isolate in Group B displayed p.a. in either control samples or in the presence of any Nx concentration. Immunoblotting revealed two positive MOR immunoreactive bands of approximately 65 and 98 kDa. MOR expression and localization was also demonstrated by immunofluorescence in isolates from Groups A and B. This study provides the first evidence of MOR expression on M. pachydermatis cell membranes pointing to its possible role in modulating p.a. production in isolates from dogs with skin lesions.
Asunto(s)
Dermatomicosis/veterinaria , Enfermedades de los Perros/microbiología , Proteínas Fúngicas/análisis , Regulación Fúngica de la Expresión Génica , Malassezia/enzimología , Fosfolipasas/biosíntesis , Receptores Opioides mu/análisis , Animales , Western Blotting , Dermatomicosis/microbiología , Perros , Técnica del Anticuerpo Fluorescente , Proteínas Fúngicas/química , Proteínas Fúngicas/fisiología , Malassezia/química , Malassezia/aislamiento & purificación , Malassezia/fisiología , Peso Molecular , Naloxona/metabolismo , Receptores Opioides mu/química , Receptores Opioides mu/fisiologíaRESUMEN
The mu-opioid receptor (MOR) is expressed in bovine, human, equine and canine oocytes, and in seasonal breeders, it is expressed with higher intensity during the anoestrous phase. Supplementation of in vitro maturation (IVM) medium with opioid agents, agonists or antagonists, was shown to affect oocyte maturation in several species such as rat, bovine and equine. This study reports the effects of supplementing IVM medium with naloxone (Nx), an opioid antagonist, on nuclear and cytoplasmic maturation rate of oocytes recovered from anoestrous bitches. Cytoplasmic maturation was examined in terms of mitochondrial (mt) distribution. In order to confirm the receptor-mediated action of Nx, in oocytes of anoestrous bitches, MOR expression was analyzed by Western blot. Cumulus-oocyte complexes, recovered from the ovaries of bitches in anoestrous, were cultured in vitro and Nx was added at the concentrations of 1 x 10(-6), 1 x 10(-8) and 1 x 10(-10) M. The rate of oocytes resuming meiosis after culture in presence of 1 x 10(-6) M Nx (29%) was significantly higher than that of oocytes of control group (12%; p < 0.05). However, treatment with Nx did not affect mt distribution pattern. In denuded oocytes and in corresponding cumulus cells, a doublet of 65 and 50 kDa was observed. We conclude that, in oocytes of anoestrous bitches, MOR is expressed and Nx significantly improves nuclear maturation rate. Further studies should be performed to elucidate the expression of other opioid receptors, such as delta and kappa, and possible interactive effects of their antagonists on canine oocyte maturation.
Asunto(s)
Regulación de la Expresión Génica/fisiología , Naloxona/farmacología , Oocitos/efectos de los fármacos , Oocitos/metabolismo , Receptores Opioides mu/metabolismo , Anestro/fisiología , Animales , Células Cultivadas , Perros , Femenino , Antagonistas de Narcóticos/farmacología , Receptores Opioides mu/genéticaRESUMEN
Equine cumulus-oocyte complexes (COCs) were analyzed by means of 13 lectins to evaluate their glycoconjugate patterns and to verify differences between COCs recovered with compact (Cp) and expanded (Exp) cumulus. Cumulus cells showed a similar staining pattern in both Cp and Exp COCs with all lectins used, except for a higher reactivity with SNA and GSA II in Cp COCs and SBA in Exp COCs. The zona pellucida (ZP) showed (1) uniform staining with MAL II, RCA(120), and SBA in both Cp and Exp COCs, (2) trilaminar binding pattern with WGA as well as higher Con A reactivity in the outer region of both types of COCs, (3) uniform staining with PNA only in Exp COCs, (4) uniform and trilaminar binding pattern with SNA in Cp and Exp COCs, respectively, and (5) major reactivity with GSA II in Exp COCs. Ooplasm showed similar staining intensity with Con A, HPA, GSA I-B(4), and WGA in both Cp and Exp COCs, with stronger reactivity to GSA II in Exp COCs, whereas SNA, UEA I, and LTA binding sites were present only in Cp COCs. Oocyte cortical granules of both Cp and Exp COCs reacted with Con A and WGA. These results suggest that, in the mare, viable (Cp) and atretic (Exp) COCs display different glycoconjugate staining pattern, which may account for the different maturation and developmental competence of COCs.
Asunto(s)
Células del Cúmulo/citología , Células del Cúmulo/metabolismo , Glicósidos/metabolismo , Caballos/fisiología , Lectinas/metabolismo , Oocitos/metabolismo , Animales , Sitios de Unión , Metabolismo de los Hidratos de Carbono/fisiología , Separación Celular , Células Cultivadas , Citoplasma/metabolismo , Femenino , Caballos/metabolismo , Oocitos/citología , Zona Pelúcida/metabolismoRESUMEN
Lipid droplets (LDs) and mitochondria in the ooplasm are essential for energy production required for maturation, fertilization and embryo development. This study investigates the correlations between cytoplasmic LDs polar aggregation and: (1) nuclear maturation (Experiment 1); (2) mitochondrial (mt) distribution pattern and localization (Experiment 2); (3) fertilization and embryonic development after intracytoplasmic sperm injection (ICSI; Experiment 3) in equine oocytes recovered from slaughtered mares and matured in vitro. Morphologically normal oocytes were selected after culture and categorized as having polar (P) aggregation or uniform (U) distribution of LDs. In Experiment 1, the maturation rate was significantly higher in P compared with U oocytes (69%, 40/58 vs. 32%, 13/41; P<0.001). In Experiment 2, it was observed that P and U oocytes showed heterogeneous mt distribution at comparable rates (68%, 25/37 vs. 50%, 2/4 for P and U respectively; NS). Moreover, only in 8/25 (32%) of P oocytes, LDs overlapped with mt aggregates in the area containing meiotic spindle. In Experiment 3, normal fertilization (51%, 19/37 vs. 60%, 6/10, for P and U) and cleavage rates (83%, 20/24 vs. 67%, 4/6, for P and U) did not differ between groups, also in oocytes with LDs located nearby the polar body. Overall, P aggregation of LDs was related to cumulus expansion at collection. In conclusion, in equine matured oocytes, P aggregation of LDs is related with cumulus expansion and nuclear maturation. However, it is not related with heterogeneous mt distribution and cannot be considered a predictive indicator for normal fertilization and embryo development.
Asunto(s)
Citoplasma/metabolismo , Caballos/embriología , Lípidos/análisis , Mitocondrias/fisiología , Oocitos/citología , Inyecciones de Esperma Intracitoplasmáticas/veterinaria , Animales , Citoplasma/química , Desarrollo Embrionario/fisiología , Fertilización/fisiología , Oocitos/fisiologíaRESUMEN
The interleukin-1 (IL-1) system is thought to be involved in periovulatory events in the mare. Previous in vivo studies have demonstrated that IL-1beta induces oocyte maturation, but depresses the pregnancy rate 14 days after ovulation. To better understand the role of IL-1 in oocyte maturation and fertilization, the effects of IL-1 on the in vitro maturation rate of equine oocytes in pure follicular fluid were evaluated and fertilization rate assessed following intracytoplasmic sperm injection (ICSI). Oocytes collected from slaughterhouse ovaries were cultured in four different media for 30 h prior to fertilization. Two experiments were performed, each using three maturation media as the experimental treatments. Medium 1 was pure follicular fluid from subordinate follicles. Medium 2 was medium 1 plus 50 ng/ml recombinant human IL-1beta. Medium 3 was pure follicular fluid collected from mares administered crude equine gonadotropin (CEG). Medium 4 was medium 2 plus 50 ng/ml of recombinant human IL-1 receptor antagonist. Media 1, 2 and 3 were compared in experiment 1. In experiment 2, media 1, 2 and 4 were compared. After maturation, metaphase II oocytes were submitted to microinjection and assessed for signs of fertilization. In experiment 1, 101 oocytes were evaluated. The rate of polar body extrusion was 66, 51 and 68% and the proportions of normally fertilized oocytes after ICSI were 40, 18 and 38% for media 1, 2 and 3, respectively. In experiment 2, 122 oocytes were evaluated. The rate of polar body extrusion was 55, 48 and 42% and the proportions showing normal fertilization after ICSI were 14, 25 and 29% for media 1, 2 and 4, respectively. There was no positive effect of IL-1beta on maturation in both experiments, but the fertilization rate and percentage of embryos reaching four-cell were low in the presence of IL-1beta, indicating that this cytokine may interfere with fertilization and early embryo development.