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Background: Chronic stress has been investigated as a risk factor for breast cancer. Maternal separation (MS) of rats has been used as a chronic stress model that alters certain systemic functions, such as the immune response. Objective: The aim of this study was to determine the possible effect of MS on the development of breast tumors induced by 7,12-dimethyl benzo anthracene (DMBA). Methodology: postnatal day (PND) 1 female Wistar rats were divided into four experimental groups that either were or were not subjected to MS and either received or did not receive DMBA. For MS, PND 1 to 21 pups were separated from their mothers for 360 min/day. On PND 30, carcinomas were induced in mammary glands using DMBA. Body weight was evaluated, and the injected region was palpated. In addition, the mammary glands were subjected to histological examination, and corticosterone levels were determined in all groups. Results: DMBA-induced groups had significantly lower body weight gain compared with the non-DMBA-induced groups. Maternal separation increased the incidence of preneoplastic changes and breast carcinogenesis in DMBA-treated animals compared with control animals. Corticosterone levels were increased in both DMBA-induced and MS groups without interaction. Conclusion: MS is a possible risk factor for DMBA-induced preneoplastic changes and breast tumors in rats.

A Zika virus (ZIKV) strain was isolated from an acute febrile patient during the Zika epidemics in Colombia. The strain was intraperitoneally inoculated into BALB/c mice, and 7 days postinoculation, neurological manifestations and ZIKV infection in the brain were demonstrated. The reported genome sequence is highly related to strains circulating in the Americas.

Objetivos: Estudiar el efecto de la infección con rabia sobre la ultraestructura dendrítica de las neuronas piramidales de la corteza cerebral en ratones inoculados con el virus por vía intramuscular. Métodos: Ratones adultos inoculados con el virus de la rabia y ratones inoculados con solución vehículo sin el virus (controles) fueron fijados por perfusión intracardiaca, con una solución que contenía paraformaldehído al 4% y glutaraldehído al 2%, cuando los animales infectados manifestaron signos avanzados de la enfermedad. Los encéfalos fueron extraídos y cortados en plano coronal en un vibrátomo. Fragmentos pequeños y delgados de estos cortes, que contenían el área de la corteza cerebral motora, fueron procesados para microscopía electrónica de transmisión. Resultados: En las dendritas distales de las neuronas piramidales de los animales controles se observaron mitocondrias largas y estrechas, así como abundantes microtúbulos organizados en paralelo con la membrana celular. En las dendritas distales de las neuronas piramidales de los ratones infectados con el virus se observaron unas estructuras electrodensas de forma irregular semejantes a figuras de mielina, pero no se observaron las mitocondrias alargadas y los microtúbulos fueron escasos. Algunas dendritas también exhibieron la formación de vacuolas que interrumpían la continuidad del citoplasma y los microtúbulos. Conclusión: La infección con virus de la rabia generó cambios ultraestructurales en las dendritas de las neuronas piramidales corticales que aparentemente no se conocían. Estos resultados son coherentes con hallazgos previos, usando otras técnicas y modelos experimentales, en donde se ha demostrado patología dendrítica inducida por la infección con rabia. Objectives: To study the effect of rabies infection on the dendritic ultrastructure of pyramidal neurons in the cerebral cortex of mice intramuscularly inoculated with rabies virus. Methods: Adult mice inoculated with rabies virus and mice inoculated with vehicle solution without the virus (controls) were fixed by intracardiac perfusion with a solution containing 4% paraformaldehyde and 2% glutaraldehyde. When infected animals showed advanced signs of disease, their brains were extracted and cut into the coronal plane on a vibratome. Small and thin fragments of these cuts containing motor cortex area were processed for transmission electron microscopy. Results: Distal dendrites of pyramidal neurons of control animals showed long and narrow mitochondria and abundant microtubules arranged in parallel with the cell membrane. In distal dendrites of pyramidal neurons of rabiesinfected mice some irregular shape electrondense structures similar to myelin figures were observed but elongated mitochondria were not observed, and microtubules were scarce. Some dendrites also exhibited vacuole formation interrupting the continuity of cytoplasm and microtubules. Conclusion: Infection with rabies virus produced ultrastructural changes within dendrites of the cortical pyramidal neurons that apparently were not known. These results are consistent with previous findings using other techniques and experimental models where it has been shown dendritic pathology induced by infection with rabies.

SUMMARY: There are few studies of infection by rabies virus in the olfactory bulb (OB). This work was carried out with the purpose of establishing the time required to detect rabies antigens in the OB of mouse, after the intramuscular inoculation of the virus and to evaluate the effect of the infection on the expression of three proteins: calbindin (CB), parvalbumin (PV) and the glial fibrillary acidic protein (GFAP). Mice were inoculated with rabies virus intramuscularly in the hind limbs. Every 8 hours, after 72 hours postinoculation (p.i.), animals were sacrificed by perfusion with paraformaldehyde and coronal sections of OB were obtained for immunohistochemical study. These cuts were used to reveal the entry and spread of viral antigens. Tissue sections obtained in the terminal phase of the disease (144 hours p.i.), and controls of the same age were also processed for immunohistochemistry of CB, PV and GFAP. Rabies virus antigens were initially detected at 80 hours p.i. in a few mitral cells. At 88 hours p.i. the antigens had spread through most of these neurons but until the terminal phase of the disease there was little dispersion of the virus towards other cellular layers of the OB. The CB protein was expressed in cells of the glomerular stratum, the PV in cells of the outer plexiform layer and the GFAP was expressed in all the layers of the OB. Viral infection generated loss of CB expression and increase of PV expression. Immunoreactivity to GFAP was increased in the outer plexiform layer of the OB as a response to infection.

Si bien lo ideal es llevar a cabo la preservación de los tejidos en el menor tiempo posible luego de la muerte de un animal objeto de un estudio neurohistoquímico, con frecuencia es inevitable trabajar con tejido nervioso obtenido varias horas post mórtem.

Resumen Introducción. Es escasa la información sobre los detalles neuroanatómicos del transporte del virus de la rabia en su ascenso por la médula espinal. Objetivos. Identificar la ruta neuroanatómica de diseminación del virus de la rabia en cada uno de los niveles de la médula espinal de ratón, después de ser inoculado por vía intramuscular. Materiales y métodos. Se inocularon ratones en los músculos isquiotibiales, con virus de la rabia. A partir de las 24 horas después de la inoculación, cada ocho horas se sacrificaron cinco animales por perfusión con paraformaldehído, se les extrajo la médula espinal y se hicieron cortes transversales en los niveles lumbosacro, torácico y cervical. Estos se procesaron mediante inmunohistoquímica para detectar antígenos virales. Resultados. Los primeros antígenos de la rabia se observaron como partículas agregadas, en la médula espinal lumbar, a las 24 horas después de la inoculación, dentro del asta ventral ipsilateral a la extremidad inoculada. A las 32 horas después de la inoculación, se hicieron visibles las primeras motoneuronas inmunorreactivas al virus. A las 40 horas después de la inoculación, se revelaron las primeras neuronas inmunorreactivas en la médula torácica, localizadas en la lámina 8 y, a las 48 horas después de la inoculación en la médula cervical, también en la lámina 8. A las 56 horas después de la inoculación, el virus se había diseminado por toda la médula espinal pero los animales aún no revelaban signos de la enfermedad. Conclusión. En el modelo de ratón aquí utilizado, el virus de la rabia ingresó a la médula espinal por las motoneuronas y, probablemente, utilizó la vía propioespinal descendente para su transporte axonal retrógrado hasta el encéfalo. Abstract Introduction: Information about the neuroanatomical details of the ascendant transport of the rabies virus through the spinal cord is scarce. Objective: To identify the neuroanatomical route of dissemination of the rabies virus at each of the levels of the spinal cord of mice after being inoculated intramuscularly. Materials and methods: Mice were inoculated with the rabies virus in the hamstrings. After 24 hours post-inoculation, every eight hours, five animals were sacrificed by perfusion with paraformaldehyde. Then, the spinal cord was removed, and transverse cuts were made at the lumbosacral, thoracic, and cervical levels. These were processed by immunohistochemistry for the detection of viral antigens. Results: The first antigens of rabies were observed as aggregated particles in the lumbar spinal cord at 24 hours post-inoculation, within the ventral horn in the same side of the inoculated limb. At 32 hours post inoculation the first motoneurons immunoreactive to the virus became visible. At 40 hours post-inoculation the first immunoreactive neurons were revealed in the thoracic level, located on lamina 8 and at 48 hours post-inoculation in the cervical cord, also on lamina 8. At 56 hours post-inoculation the virus had spread throughout the spinal cord, but the animals still did not show signs of the disease. Conclusion: In the mouse model we used, the rabies virus entered the spinal cord through the motoneurons and probably used the descending propriospinal pathway for its retrograde axonal transport to the encephalus.

Previous studies using the Golgi technique have demonstrated alterations in the dendritic morphology of pyramidal neurons of the cerebral cortex of mice inoculated with the rabies virus. However, knowledge about the fine structure of dendrites in rabies infection is scarce. This work had the aim of studying the ultrastructure of dendrites in cortical pyramidal neurons of rabies-infected mice. Mice were inoculated intramuscularly with a street rabies virus of canine origin. The animals that showed an advanced stage of disease were fixed by perfusion with glutaraldehyde and paraformaldehyde. Brains were removed and cut on a vibratome to obtain coronal slices of 200 micrometers of thickness. Vibratome slices were subjected to the following treatment: postfixation, dehydration, embedding in epoxy resin and polymerization between glass slides. Ultrathin sections of oriented tissue fragments from the cerebral cortex were obtained and observed under electron microscope. The most significant ultrastructural findings were located within distal dendrites of cortical pyramidal neurons: loss of mitochondria, disorganization and loss of microtubules, formation of vacuoles interrupting the continuity of the cytoplasm and formation of myelin-like figures. These strange myelin figures, which apparently had not been reported in previous studies of rabies, were the most noticeable ultrastructural feature. They also differ from the best known myelin figures formed by concentric lamellae. The possible origin of these myelin figures as result of mitochondrial degeneration is discussed.

Rabies is a fatal infectious disease of the nervous system; however, the knowledge about the pathogenic neural mechanisms in rabies is scarce. In addition, there are few studies of rabies pathology of the spinal cord.To study the distribution of calcium binding proteins calbindin and parvalbumin and assessing the effect of rabies virus infection on their expression in the spinal cord of mice. MATERIALES Y METHODS: Mice were inoculated with rabies virus, by intracerebral or intramuscular route. The spinal cord was extracted to perform some crosscuts which were treated by immunohistochemistry with monoclonal antibodies to reveal the presence of the two proteins in normal and rabies infected mice. We did qualitative and quantitative analyses of the immunoreactivity of the two proteins.Calbindin and parvalbumin showed differential distribution in Rexed laminae. Rabies infection produced a decrease in the expression of calbindin. On the contrary, the infection caused an increased expression of parvalbumin. The effect of rabies infection on the two proteins expression was similar when comparing both routes of inoculation.The differential effect of rabies virus infection on the expression of calbindin and parvalbumin in the spinal cord of mice was similar to that previously reported for brain areas. This result suggests uniformity in the response to rabies infection throughout the central nervous system. This is an important contribution to the understanding of the pathogenesis of rabies.