Bovine herpesvirus type 5 infection regulates Bax/BCL-2 ratio.

Bovine herpesvirus 5 (BoHV-5) is an α-herpesvirus that causes neurological disease in young cattle and is also occasionally involved in reproductive disorders. Although there have been many studies of the apoptotic pathways induced by viruses belonging to the family Herpesviridae, there is little information about the intrinsic programmed cell death pathway in host-BoHV-5 interactions. We found that BoHV-5 is able to replicate in both mesenchymal and epithelial cell lines, provoking cytopathology that is characterized by cellular swelling and cell fusion. Viral antigens were detected in infected cells by immunofluorescence assay at 48 to 96 h post-infection (p.i.). At 48 to 72 h p.i., anti-apoptotic BCL-2 antigens were found at higher levels than Bax antigens; the latter is considered a pro-apoptotic protein. Infected cells had increased BCL-2 phenotype cells from 48 to 96 h p.i., based on flow cytometric analysis. At 48 to 96 h p.i., Bax mRNA was not expressed in any of the infected cell monolayers. In contrast, BCL-2 mRNA was found at high levels at all p.i. in both types of cells. BoHV-5 replication apparently modulates BCL-2 expression and gene transcription, enhancing production of virus progeny.

Immunohistochemical and molecular analysis of caveolin-1 expression in canine mammary tumors.

Caveolin-1 (Cav-1) is a structural protein present in invaginations of the cell membrane. In human breast cancer, the cav-1 gene is believed to be a tumor suppressor gene associated with inhibition of tumor metastasis. However, little is known about its expression, regulation and function in canine mammary tumors. Expression levels of cav-1 were investigated using real-time PCR and immunohistochemical detection with an anti-human Cav-1 antibody. Gene expression stability of different samples was analyzed using the geNorm software. Mammary tumors from 51 female dogs were compared to normal mammary tissue from 10 female dogs. Malignant mammary cells showed a loss of Cav-1 expression by quantitative RT-PCR and weak Cav-1 staining by immunohistochemistry compared to normal mammary gland tissue. There was a significant relationship between outcome and immunostaining as well as with tumor size, indicating that caveolin subexpression has a positive predictive value and is related to higher survival and smaller tumor size. Our findings indicate that Cav-1 is a potential prognostic marker for canine mammary tumors.

Apoptotic and developmental effects of bovine Herpesvirus type-5 infection on in vitro-produced bovine embryos.

Bovine Herpesvirus type-5 (BoHV-5), which is potentially neuropathogenic, was recently described to be related with reproductive disorders in cows. The objective was to elucidate mechanisms involved in propagation of BoHV-5 in embryonic cells. For this purpose, bovine embryos produced in vitro were assayed for apoptotic markers after experimental infection of oocytes, in vitro fertilization, and development. Host DNA fragmentation was detected with a TUNEL assay, expression of annexin-V was measured with indirect immunofluorescence, and viral DNA was detected with in situ hybridization. Infective BoHV-5 virus was recovered from embryos derived from exposed oocytes after two consecutive passages on Madin-Darby bovine kidney (MDBK) cells. The viral DNA corresponding to US9 gene, localized between nucleotides 126243 to 126493, was detected in situ and amplified. There was no significant difference between the ratio of TUNEL stained nuclei and total cells in good quality blastocysts (0.87 ± 0.05, mean ± SD), but there were differences (P < 0.05) between infected (0.18 ± 0.05) and uninfected blastocysts (0.73 ± 0.07). The Annexin-V label was more intense in uninfected embryos (0.79 ± 0.04; P < 0.05). The quality of infected and uninfected embryos was considered equal, with no significant effect on embryonic development. In conclusion, we inferred that BoHV-5 infected bovine oocytes, replicated, and suppressed some apoptotic pathways, without significantly affecting embryonic development.

Effects of bovine Herpesvirus Type 5 on development of in vitro-produced bovine embryos.

Bovine (Bos indicus) herpesviruses have been associated with reproductive disease. Type 1, the most studied species, is best known for its reproductive and respiratory effects. Type 5 (BoHV-5) has been detected in bull semen and aborted fetuses but not in oocytes and embryos. This study consisted of three experiments that evaluated (1) BoHV-5-infected oocytes matured in medium with fetal bovine serum (BoHV-FBS) or polyvinyl alcohol (BoHV-PVA) and fertilized by noninfected sperm; (2) noninfected oocytes fertilized by BoHV-5-infected sperm; and (3) infection of presumptive zygotes by BoHV-5. Each treatment involved nine drops of 15 to 20 oocytes. Infection with BoHV-5 was detected by polymerase chain reaction and in situ hybridization assay, and fertilization capacity and embryonic development were assessed using in vitro culture. Experimentally induced infection was obtained in all experiments, and vertical transmission of BoHV-5 by gametes was confirmed. The cleavage rate was reduced (P=0.0201) in BoHV-FBS (80.4+/-8.9%; mean+/-SD) compared with that of noninfected oocytes (89.9+/-6.5%); neither differed from BoHV-PVA (87.3+/-7.1%), and the resulting embryo production rate was not significantly different among groups. Rates of cleavage (87.5+/-7.5% vs. 92.2+/-5.5%, control vs. infected) and development of embryos (41.7+/-9.9% vs. 44.3+/-7.7% to morula/blastocyst/expanded blastocyst [M/B/EB] and 39.6+/-10.3% vs. 40.8+/-9.2% to blastocyst/expanded blastocyst/hatching blastocyst [B/EB/HB] stages) were not compromised by infected sperm (P=0.1462, P=0.5402, and P=0.8074, respectively). However, presumptive zygotes directly infected 1 d after fertilization produced a lower number (P=0.0140 to M/B/EB and P=0.002 to B/EB/HB stages) of in vitro-produced embryos (31.6+/-4.6 vs. 25.0+/-5.5 and 31.6+/-4.6 vs. 20.2+/-5.4; control vs. infected). In conclusion, BoHV-5 infected gametes and was transmissible to the embryo during in vitro development. As zygotes infected 1 d after fertilization had compromised development, BoHV-5 has the potential to be a pathogen with economic consequences.

Actividad citotoxica de plantas medicinales usadas por el grupo étnico boliviano Tacana / Cytotoxic activity of medicinal plants used by the Takana ethnic group in Bolivia

Pregunta de investigación ¿ Las plantas utilizadas en la Medicina Tradicional por los Tacanas manifiestan actividad citotóxica in vitro con el test de la sulforodamina? Objetivos. Determinar la actividad citotóxica in vitro de extractos de plantas utilizadas en la Medicina Tradicional por los Tacanas. Lugar. Población Tacana, IBBA. Métodos. Cultivo de lineas celulares humanas, KB (carcinoma nasofaringeo), HeLa (carcinoma cervical)Ensayo colorimétrico de sulforodamina B, in vitro. Resultados. 118 extractos etanólicos de especies vegetales estudiadas in vitro utilizadando dos lineas tumorales humanas. 14 especies vegetales mostraron actividad citotóxica in vitro. De la 14 especies citotóxicas, 12 especies vegetales presentaron Cl50<- a 10ug/ml. Conclusión. 13 porciento de las plantas estudiadas presentaron actividad citotóxica, pese a no ser seleccionadas por su uso tradicional como anticancerígenos, pues, la actividad anticancer de las plantas estudiadas no consta en la medicina tradicional de los Tacanas. La actividad antitumoral, antioxidante y antimutagénica dae las especies citotóxicas en este trabajo y otras especies diferentes, per, de los mismos géneros estudiados fue reportada por científicos de otras latitudes, lo que nos podría indicar que los principios activos serian comunes en estos géneros, permitiéndonos indentificar nuevas fuentes de medicamentos. Todas las especies activas merecen estudios complementarios.

[Descriptive epidemiological study of a year-long otologic consultation in a tertiary hospital]. / Estudio epidemiológico descriptivo de una consulta de otología durante un año en un hospital de tercer nivel.

Despite the spreading of Otologic care Units it is surprising the lack of comprehensive epidemiologic descriptions of patients attended in these Departments. A greater efficacy would be the result of better protocols designs and consequently a much better sanitary behaviour. We make a register of protocols and several aspects epidemiologic and diagnostical of our first consultations in the Unit during 1998. Were studied 595 cases, 320 women and 275 men, middle age was 49.91 years. From each one was controlled and analyzed the age, the sex, where they come from under the sanitary view-point, consult reason and diagnosis achieved.

[Analysis of the resonant frequency of the middle ear in otosclerosis]. / Análisis de la frecuencia de resonancia del oído medio en la otosclerosis.

Multifrequency tympanometry scans pressures and frequencies from 226 to 2000 Hz to analyze the resonant frequency of the middle ear, which determines the balance between stiffness and mass. We studied 143 patients (a total of 200 ears) diagnosed as otosclerosis at the E.N.T. Service of Santiago de Compostela University Hospital Complex, and 136 control subjects. The mean age of patients was 47.7 years; women were predominant (female/male ratio 2.25:1). The mean duration of the disease was 10.4 years. Using the Virtual model 310 tympanometer, we calculated the resonant frequency through the susceptance tympanogram, corresponding to the minimal frequency at which the central notch of the tympanograrn was equal to or less than the negative tail. The mean resonant frequency was 1230 Hz, the tympanometric amplitude was 88.32 daPa, and the admittance of the middle ear was 0.63 cc. We confirmed an increase in the resonant frequency in patients with otosclerosis compared with normal subjects (1132.33 Hz), and a decrease in the admittance of the middle ear. The relation between the two values was significant.

[Parameters of normality in multifrequency tympanometry]. / Parámetros de normalidad en la timpanometría de multifrecuencia.

Multifrequency tympanometry studies consist of tympanography using probe tone frequencies ranging from 200 to 2000 Hz, improving the study of acoustic transmission through the tympano-ossicular system because then two components of admittance, conductance and susceptance, can be separated. The resonance frequency is the frequency at which mass and spring elements of the middle ear cancel each other out, leaving only the friction component. This measurement has been found to be more sensitive to the presence of pathologies that affect the tympano-ossicular system, such as otosclerosis and rheumatoid arthritis. It is necessary to know normal pattents of tympanometric parameters to improve the study of these diseases. Multifrequency tympanometry performed on 136 patients, 91 women and 45 men, age range 11-78 years. The mean resonant frequency of the middle ear was 1132.33 Hz, mean static admittance 0.76 dapa, and mean tympanometric amplitude 94.31 mmhos ac. Age showed no systematic effect of age on any of these measures in this population, and no significant association was found between static admittance or tympanometric amplitude and resonance frequency.

Análisis de la frecuencia de resonancia del oído medio en la otosclerosis / Analysis of the resonant frequency of the middle ear in otosclerosis

La timpanometría de multifrecuencia realiza un barrido de presiones y de frecuencias desde los 226 Hz a los 2. 000 Hz, permitiendo analizar la frecuencia de resonancia del oído medio, que determina el equilibrio entre la rigidez y la masa del mismo. Hemos estudiado 143 pacientes diagnosticados de otosclerosis en el Servicio de Otorrinolaringología del Complejo Hospitalario Universitario de Santiago de Compostela, con un total de 200 oídos estudiados, así como 136 personas normales como controles. La edad media de los pacientes al diagnóstico fue de 47,71 años, con un predominio del sexo femenino (relación mujer/varón de 2,25: 1), con una duración media de la enfermedad de 10,4 años. Utilizamos el timpanómetro Virtual modelo 310, calculando lafrecuencia de resonancia a través del timpanograma de susceptancia, correspondiendo a la mínima frecuencia en la que la muesca central del timpanograma es igual o inferior al extremo negativo del mismo. De esta forma, la frecuencia media de resonancia fue de 1.230 Hz, la amplitud timpanométrica de 88,32 daPa y la admitancia del oído medio de 0,63 cc. Hemos comprobado un incremento de la frecuencia de resonancia en los pacientes con otosclerosis, al compararlos con los valores de normalidad (1.132,33 Hz), así como una disminución de la admitancia del oído medio, existiendo entre los dos valores una relación significativa (AU)

Multifrequency tympanometry scans pressures and frequencies from 226 to 2000 Hz to analyze the resonant frequency of the middle ear, which determines the balance between stiffness and mass. We studied 143 patients (a total of 200 ears) diagnosed as otosclerosis at the E.N.T. Service of Santiago de Compostela University Hospital Complex, and 136 control subjects. The mean age of patients was 47.7 years; women were predominant (female/male ratio 2.25:1). The mean duration of the disease was 10.4 years. Using the Virtual model 310 tympanometer, we calculated the resonant frequency through the susceptance tympanogram, corresponding to the minimal frequency at which the central notch of the tympanograrn was equal to or less than the negative tail. The mean resonant frequency was 1230 Hz, the tympanometric amplitude was 88.32 daPa, and the admittance of the middle ear was 0.63 cc. We confirmed an increase in the resonant frequency in patients with otosclerosis compared with normal subjects (1132.33 Hz), and a decrease in the admittance of the middle ear. The relation between the two values was significant (AU)

Parámetros de norma lidad en la timpanometría de multifrecuencia / Parameters of normality in multifrequency tympanometry

La timpanometría de multifrecuencia realiza un barrido timpanométrico entre 226 y 2000 Hz, mejorando el análisis de la transmisión del sonido a través del sistema tímpano-osicular, ya que posibilita el estudio independiente de los dos componentes de la admitancia del oído medio, la susceptancia y la conductancia. La frecuencia de resonancia del oído medio es la frecuencia en la que los elementos distensibles y de masa de la caja timpánica están en equilibrio; por tanto este parámetro se muestra útil en el estudio de patologías que alteren las características del sistema tímpano osicular como la otosclerosis y la artritis reumatoide. Es preciso conocer los parámetros de normalidad con el objeto de poderlos comparar con estas patologías. Realizamos la timpanometría de multifrecuencia a 136 pacientes entre 11 y 78 años. La frecuencia de resonancia media de oído medio fue de 1132.33 Hz, la admitancia estática media de 0, 76 dapa, y la amplitud timpanométrica media de 94,31 nmohs ac. Ninguno de estos parámetros mostró variaciones significativas en los distintos grupos de edad, y tampoco observamos asociación entre la frecuencia de resonancia de oído medio y la admitancia estática o la amplitud timpanométrica (AU)

Multifrequency tympanometry studies consist of tympanography using probe tone frequencies ranging from 200 to 2000 Hz, improving the study of acoustic transmission through the tympano-ossicular system because then two components of admittance, conductance and susceptance, can be separated. The resonance frequency is the frequency at which mass and spring elements of the middle ear cancel each other out, leaving only the friction component. This measurement has been found to be more sensitive to the presence of pathologies that affect the tympano-ossicular system, such as otosclerosis and rheumatoid arthritis. It is necessary to know normal pattents of tympanometric parameters to improve the study of these diseases. Multifrequency tympanometry performed on 136 patients, 91 women and 45 men, age range 11-78 years. The mean resonant frequency of the middle ear was 1132.33 Hz, mean static admittance 0.76 dapa, and mean tympanometric amplitude 94.31 mmhos ac. Age showed no systematic effect of age on any of these measures in this population, and no significant association was found between static admittance or tympanometric amplitude and resonance frequency (AU)

Diagnostic value of the multifrequency tympanometry in active rheumatoid arthritis.

We carried out a functional evaluation of the malleus-incus and incus-stapes joints in 37 patients with rheumatoid arthritis using multifrequency tympanometry. As a reference point, we calculated the resonance point using two different methods, and the results obtained were classified into two groups according to the phase of the disease: active, with clinical affection of the joint and with no medication; and inactive, asymptomatic. The results were compared with previously published criteria indicating normality, which underlined a significant correlation in the analysis of the resonance point between both groups, and which determined the value of multifrequency tympanometry in that stage and reactivation of the rheumatoid arthritis in active phase.