Extensive burn injury causes bone collagen network alteration and growth delay related to RANK-L immunoexpression change.

PURPOSE: Extensive burn injury mainly affects children, and hypermetabolic state can lead to growth delay. This study aimed to investigate bone histopathological and morphometric aspects, collagen fibers network and the immunoexpression of biological markers related to bone development in a young experimental model for extensive burn. MATERIALS AND METHODS: A total of 28 male Wistar rats were distributed into Control (C) and subjected to scald burn injury (SBI) groups. Sham or injured animals were euthanized 4 or 14 days post-lesion and proximal epiphyses of the femur were submitted to histological, morphometric (thickness epiphyseal plate), and RUNX-2 and receptor activator of nuclear factor kappa- ß ligand (RANK-L) immunoexpression methods. RESULTS: Histopathological femoral findings showed delayed appearance of the secondary ossification center in SBI, 14 days post-injury. Collagen fibers 4 days after injury were observed in articular cartilage as a pantographic network with a transversally oriented lozenge-shaped mesh, but this network was thinner in SBI. Fourteen days after the injury, the pantographic network of collagen presented square-shaped mesh in C, but this aspect was changed to a wider mesh in SBI. Morphometric analysis of epiphyseal plate revealed that the SBI group had less thickness than the respective controls (p<0.05). RUNX-2 showed no difference between groups, but RANK-L score was higher in all SBI groups. CONCLUSIONS: Extensive burn injury causes delayed bone growth and morphological changes. Alterations in collagen network and enhancement in immunoreactivity of RANK-L result in increased osteoclastogenesis.

The sphenoidal emissary foramen and the emissary vein: Anatomy and clinical relevance.

Although the sphenoidal emissary foramen (SEF) and its content are anatomically and clinically relevant, accurate description of them in the modern literature is lacking. This study aimed to examine and describe the SEF and its content (the sphenoidal emissary vein [SEV]). We analyzed 1,000 computed tomography (CT) images, 170 dry skulls, 50 formalin-fixed specimens, and three specimens (heads) following guidelines proposed by Dr. Albert L. Rhoton Jr. MD for latex injection. SEV morphology was determined by histological staining and electron microscopy. The SEF was observed in 46.8% of the CTs studied (25.4% bilateral and 21.4% unilateral), and 45.2% of the dry skulls (18.8% bilateral and 26.4% unilateral). In 9.5% of CTs and 21.1% of dry skulls there was a blind channel in the external surface of the cranial base; since there was no communication with the cranial cavity, it was not considered as the SEF. During the dissections, the SEF was found in seven individuals. In three of them, the SEV was an alternative route for venous drainage of the venous plexus of the foramen ovale. Its walls were composed of collagen fibers and its endothelium contained rhomboid cells resembling those commonly found in the superior sagittal sinus. The presence of the SEF and SEV can anatomically explain the spread of certain cranial base pathologies from or toward Meckel's cave or the cavernous sinus, and should be taken into account during procedures in the middle cranial fossa, percutaneous approaches, odontological procedures, and treatment of dural arteriovenous fistulas. Clin. Anat., 33:767-781, 2020. © 2019 Wiley Periodicals, Inc.

Temporal study following burn injury in young rats is associated with skeletal muscle atrophy, inflammation and altered myogenic regulatory factors.

INTRODUCTION: Burn injury (BI) greater than 40% has been associated with protein catabolism and it is characterized by a hypermetabolic response followed for muscle loss. OBJECTIVE: The purpose of this study was to investigate the temporal effects of extensive experimental BI in the skeletal muscle distant from lesion, through morphological analysis, expression of genes related to muscle atrophy, inflammation and the myogenic regulatory factors. MATERIALS AND METHODS: A total of 60 young male wistar rats were distributed into two groups, control (C) and subjected to scald burn injury (SBI). The animals were euthanized 1, 4 and 14 days post-sham or 45% of the total body surface BI. The medial head of gastrocnemii muscles were submitted to histopathological, morphometric (muscle fibers area and density), MyoD and myogenin immunoexpression, and gene expression for TNF-α, iNOS and E3 ubiquitin ligases (MuRF1 and MAFbx). RESULTS: Histopathological findings were consistent with increased amount of connective tissue and inflammatory process. Muscle fiber area of SBI groups was smaller than C and no differences were found in fiber muscle density. TNF-α was higher in SBI groups, one and 14 days post-injury; iNOS expression was higher on the first and fourth day post-injury. MuRF-1 was higher on the day four and MAFbx on the day 14. CONCLUSION: In conclusion, BI causes inflammation, atrophy and myogenesis stimulation in muscle as a result of systemic host response.

Cyclooxygenase-2 expression in skeletal muscle of knockout mice suffering Duchenne muscular dystrophy.

The purpose of the present study was to investigate the role of cyclooxygenase-2 (COX-2) expression in fibrotic lesion in mdx mice. A total of six male C57BL/10 mice and six C57BL/10-DMD/mdx were distributed into two groups: control and animals with Duchenne muscular dystrophy (DMD). The medial part of gastrocnemius muscle was evaluated being the specimens stained with hematoxylin and eosin (H&E) and Sirius Red under normal and polarized light to differentiate type I (red and yellow) and III (green) collagen. COX-2 expression was assessed by immunohistochemistry. The results revealed histopathological changes in C57BL/10-DMD/mdx as depicted by regenerating fibers. Sirius Red stain showed a substantial increase in the amount of type I collagen of mdx mice. DMD induced a strong COX-2 immunoexpression in intercellular space. Taken together, our results are consistent with the notion that necrotic and fibrotic lesions are able to increase COX-2 expression in DMD.

Effects of ischemia and reperfusion on P2X2 receptor expressing neurons of the rat ileum enteric nervous system.

PURPOSE: We investigated the effects of ischemia/reperfusion in the intestine (I/R-i) on purine receptor P2X2-immunoreactive (IR) neurons of the rat ileum. METHODS: The superior mesenteric artery was occluded for 45 min with an atraumatic vascular clamp and animals were sacrificed 4 h later. Neurons of the myenteric and submucosal plexuses were evaluated for immunoreactivity against the P2X2 receptor, nitric oxide synthase (NOS), choline acetyl transferase (ChAT), calbindin, and calretinin. RESULTS: Following I/R-i, we observed a decrease in P2X2 receptor immunoreactivity in the cytoplasm and surface membranes of neurons of the myenteric and submucosal plexuses. These studies also revealed an absence of calbindin-positive neurons in the I/R-i group. In addition, the colocalization of the P2X2 receptor with NOS, ChAT, and calretinin immunoreactivity in the myenteric plexus was decreased following I/R-i. Likewise, the colocalization between P2X2 and calretinin in neurons of the submucosal plexus was also reduced. In the I/R-i group, there was a 55.8% decrease in the density of neurons immunoreactive (IR) for the P2X2 receptor, a 26.4% reduction in NOS-IR neuron, a 25% reduction in ChAT-IR neuron, and a 47% reduction in calretinin-IR neuron. The density of P2X2 receptor and calretinin-IR neurons also decreased in the submucosal plexus of the I/R-i group. In the myenteric plexus, P2X2-IR, NOS-IR, ChAT-IR and calretinin-IR neurons were reduced in size by 50%, 49.7%, 42%, and 33%, respectively, in the I/R-i group; in the submucosal plexus, P2X2-IR and calretinin-IR neurons were reduced in size by 56% and 72.6%, respectively. CONCLUSIONS: These data demonstrate that ischemia/reperfusion of the intestine affects the expression of the P2X2 receptor in neurons of the myenteric and submucosal plexus, as well as density and size of neurons in this population. Our findings indicate that I/R-i induces changes in P2X2-IR enteric neurons that could result in alterations in intestinal motility.

The connective tissue of the adductor canal--a morphological study in fetal and adult specimens.

The adductor canal is a conical or pyramid-shaped pathway that contains the femoral vessels, saphenous nerve and a varying amount of fibrous tissue. It is involved in adductor canal syndrome, a claudication syndrome involving young individuals. Our objective was to study modifications induced by aging on the connective tissue and to correlate them to the proposed pathophysiological mechanism. The bilateral adductor canals and femoral vessels of four adult and five fetal specimens were removed en bloc and analyzed. Sections 12 microm thick were obtained and the connective tissue studied with Sirius Red, Verhoeff, Weigert and Azo stains. Scanning electron microscopy (SEM) photomicrographs of the surfaces of each adductor canal were also analyzed. Findings were homogeneous inside each group. The connective tissue of the canal was continuous with the outer layer of the vessels in both groups. The pattern of concentric, thick collagen type I bundles in fetal specimens was replaced by a diffuse network of compact collagen bundles with several transversal fibers and an impressive content of collagen III fibers. Elastic fibers in adults were not concentrated in the thick bundles but dispersed in line with the transversal fiber system. A dynamic compression mechanism with or without an evident constricting fibrous band has been proposed previously for adductor canal syndrome, possibly involving the connective tissue inside the canal. The vessels may not slide freely during movement. These age-related modifications in normal individuals may represent necessary conditions for this syndrome to develop.

Topical Insulin Modulates Inflammatory and Proliferative Phases of Burn-Wound Healing in Diabetes-Induced Rats.

The healing time of burn wounds depends on surface area and depth of the burn and associated comorbidities. Diabetes mellitus (DM) causes delays in the healing process by extending the inflammatory phase. Treatment with topical insulin can improve the inflammatory phase, restore metabolic dysregulation, and modulate impaired cellular signaling in burn wounds. The objective of this study was to evaluate markers of the inflammatory and proliferative phases of second-degree burns after topical insulin treatment in diabetic rats. Type I DM was induced with streptozotocin in male Wistar rats. The animals' backs were shaved and subjected to thermal burning. Rats were randomized into two groups: control diabetic (DC) and insulin diabetic (DI). At Days 7 and 14 postburn, rats were euthanized, and wound-tissue sections were evaluated by hematoxylin and eosin, Weigert, and Verhöeff staining, immunohistochemistry-paraffin, and enzyme-linked immunosorbent assay. A significant increase in reepithelialization was seen on Days 7 and 14 in DI versus DC rats. On Day 7, interleukin (IL)-1ß, IL-6, monocyte chemotactic protein (MCP)-1, and F4/80 expression were increased in DI versus DC rats. On Day 14, MCP-1 expression was decreased and F4/80 increased in DI versus DC rats. On Days 7 and 14, Ki-67, transforming growth factor-ß1, vascular endothelial growth factor expression, and formation of elastic fibers were increased in DI versus DC rats. Topical insulin modulates burn-wound healing in diabetic animals by balancing inflammation and promoting angiogenesis and formation of elastic fibers.

Topographic and microscopic anatomical description of the emissary sinus of foramen ovale in adult humans.

OBJECTIVE: Although the Emissary Sinus of Foramen Ovale (ESFO) was first described by Trolard in 1868, its definition remains confused and neglected in the medical literature. This structure represents a vein, two veins, a venous plexus, or a dural sinus? Does it really exist? To understand this topic, this work aimed to describe the anatomy, topography, and microscopic features of the ESFO, precisely characterizing its structure, routes and anatomical correlations. PATIENTS AND METHODS: ESFO from the skull's base of adults were dissected into fifty anatomical blocks and evaluated using Hematoxylin and Eosin, Picro-sirius red and Weigert staining, and by Scanning Electron Microscopy (SEM). RESULTS: ESFO was always present between cavernous sinus and pterygoid plexus on both antimeres, its inferior route passing through the foramen ovale and/or sphenoidal emissary foramen (foramen of Vesalius), anterior to the mandibular branch of trigeminal nerve. Its microscopic arrangement resembled what was found on transverse sinus, that is composed by layers of collagen fibers oriented on transversal and longitudinal planes. It wasn't possible to identify the media and adventitial tunica, features seen in veins, and the elastic layer was very thin near its lumen. SEM analysis showed that, like the transverse sinus, the ESFO was composed by parallel cells that presented a rhombus shape containing central rounded nuclei. CONCLUSION: In summary, the venous channel passing through the foramen ovale and/or sphenoidal emissary foramen (foramen of Vesalius) is a dural venous sinus constituted by dura mater layers and should be considered during surgical approaches near the foramen ovale in the middle cranial fossa.

Human Anatomy in Cinema / La Anatomía Humana en el Cine

SUMMARY: A corpse is the body's final destination and, as a social construction, it challenges Western cultural taboos and practices around the corpse, having been shown in cinema. The present paper aims to analyze 17 films that portray Human Anatomy, identifying which themes specifically related to morphological sciences are addressed, namely: a) reactions to first contact with a cadaver, b) medical school and dissection classes, c) origin of cadavers, d) anatomy and suspense or horror movies, e) romance, drama and comedy, f) body snatchers/ resurrectionists, g) anatomical techniques, h) racism, prejudice and anatomy. This is a critical review of films in which, after thorough triple- blind analysis by the authors, it was observed that the encounter with the cadaver provides a series of emotions to the characters participating in the practical anatomy classes shown in the films, such as apprehension, fear, disgust, curiosity, etc. Generally, this surprising contact between the living and the dead is associated with the practice of dissection, which often results in the accentuation of emotions previously reported. From all this, it is concluded that the history of the teaching of Human Anatomy has been interestingly portrayed in cinema from different points of view.

Un cadáver es el destino final del cuerpo y, como construcción social, desafía los tabúes y prácticas culturales occidentales en torno al cadáver, habiendo sido mostrado en el cine. El presente trabajo tuvo como objetivo analizar 17 películas que retratan la Anatomía Humana, identificando qué temas específicamente relacionados con las ciencias morfológicas se abordan, a saber: a) reacciones al primer contacto con un cadáver, b) clases de medicina y disección, c) origen de los cadáveres, d) anatomía y películas de suspenso o terror, e) romance, drama y comedia, f) ladrones de cuerpos/resurreccionistas, g) técnicas anatómicas, h) racismo, prejuicio y anatomía. Se trata de una revisión crítica de películas en las que, tras un minucioso análisis triple ciego por parte de los autores, se observa que el encuentro con el cadáver proporciona una serie de emociones a los personajes que participan en las clases prácticas de anatomía que se muestran en las películas, tales como aprensión, miedo, repugnancia, curiosidad, etc. Generalmente, este sorprendente contacto entre vivos y muertos se asocia con la práctica de la disección, lo que muchas veces resulta en la acentuación de las emociones anteriormente relatadas. De todo ello se concluye que la historia de la enseñanza de la Anatomía Humana ha sido interesantemente retratada en el cine desde diferentes puntos de vista.

Cardiac corpuscles: a “new” morphofunctional entity?

he existence of “transitional muscular structures” between subendocardial branches (Purkinje fibers) and ventricular working muscle fibers (WF) was first described by the German anatomist, Kurt Goerttler, in 1964. He designated them as “subendocardial nucleus organs.” He supposed such fibers functioned as mechanoreceptors, controlling of the intensity of contraction of the ventricular musculature. Brazilian anatomist Ferraz de Carvalho described similar structures in 1993. A thorough literature search failed to identify any other research articles confirming or denying their existence. The objective of this work was to find such structures in subendocardial ventricular walls in human hearts. We collected fifteen formalin-preserved hearts from the Anatomy Department of São Paulo University and sectioned the apical portions on the right and left ventricles according to method used by Goerttler. We utilized conventional histology (light microscopy- LM), scanning electron microscopy (SEM), and a new preservation method called micro- plastination (MP). At the anterior wall of the right ventricle in the subendocardial region between the interventricular septum and moderator band, we found several bundles of fusiform and helicoidal fibers of similar histology to the WF. The bundles measured between 400 and 1150 µm in length and were separated from adjacent muscular fibers by thin collagen fiber, thus acting as a “pseudo capsule.” Some structures seemed to be linked to PF and were appeared to be lymphatic and blood vessels and nerves. We called those structures “cardiac corpuscles” (CC). The observation of the previously “unknown” CC in this initial study confirmed the previous descriptions and its discovery may contribute to new perspectives in the study of cardiac muscle structure and function.

Prototyping and 3D printing of computed tomography images with an emphasis on soft tissues, especially muscles, for teaching human anatomy / Prototipado e impresión 3D de imágenes de tomografía computarizada con énfasis en los tejidos blandos, especialmente músculos, para la enseñanza de la anatomía humana

The study on cadavers, although considered fundamental in the teaching of human anatomy, is limited in several universities, mainly due to the acquisition and manipulation of cadaveric material. Throughout history, several artificial anatomical models have been used to complement the real anatomical pieces. The present study offers a new alternative: the making of three-dimensional models from Computed Tomography (3D-CT) patient image acquisition. CT images from the USP University Hospital database were used. Patients underwent examinations for reasons other than the present study and were anonymized to maintain confidentiality. The CT slices obtained in thin cross-sections (approximately 1.0 mm thick) were converted into three-dimensional images by a technique named Volume Rendering for visualization of soft tissue and bone. The reconstructions were then converted to an STL (Standard Triangle Language) model and printed through two printers (LONGER LK4 Pro® and Sethi S3®), using PLA and ABS filaments. The 3D impressions of the thigh and leg muscles obtained better visual quality, being able to readily identify the local musculature. The images of the face, heart, and head bones, although easily identifiable, although seemed to present lower quality aesthetic results. This pilot study may be one of the first to perform 3D impressions of images from CT to visualize the musculature in Brazil and may become an additional tool for teaching.

Cardiac Corpuscles: A "New" Morphofunctional Entity? / Corpúsculos Cardíacos: ¿Una "Nueva" Entidad Morfofuncional?

SUMMARY: The existence of "transitional muscular structures" between subendocardial branches (Purkinje fibers) and ventricular working muscle fibers (WF) was first described by the German anatomist, Kurt Goerttler, in 1964. He designated them as "subendocardial nucleus organs." He supposed such fibers functioned as mechanoreceptors, controlling of the intensity of contraction of the ventricular musculature. Brazilian anatomist Ferraz de Carvalho described similar structures in 1993. A thorough literature search failed to identify any other research articles confirming or denying their existence. The objective of this work was to find such structures in subendocardial ventricular walls in human hearts. We collected fifteen formalin-preserved hearts from the Anatomy Department of São Paulo University and sectioned the apical portions on the right and left ventricles according to method used by Goerttler. We utilized conventional histology (light microscopy- LM), scanning electron microscopy (SEM), and a new preservation method called micro- plastination (MP). At the anterior wall of the right ventricle in the subendocardial region between the interventricular septum and moderator band, we found several bundles of fusiform and helicoidal fibers of similar histology to the WF. The bundles measured between 400 and 1150 µm in length and were separated from adjacent muscular fibers by thin collagen fiber, thus acting as a "pseudo capsule." Some structures seemed to be linked to PF and were appeared to be lymphatic and blood vessels and nerves. We called those structures "cardiac corpuscles" (CC). The observation of the previously "unknown" CC in this initial study confirmed the previous descriptions and its discovery may contribute to new perspectives in the study of cardiac muscle structure and function.

La existencia de "estructuras musculares de transición" entre los ramos subendocárdicos (fibras de Purkinje) y las fibras musculares ventriculares activas(FMV) fue descrita por primera vez por el anatomista alemán Kurt Goerttler en 1964, quien las denominó "órganos del núcleo subendocárdico". Supuso que tales fibras funcionaban como mecanoreceptores, controlando la intensidad de la contracción de la musculatura ventricular. El anatomista brasileño Ferraz de Carvalho describió estructuras similares en 1993. Una búsqueda bibliográfica exhaustiva no logró identificar ningún otro artículo de investigación que confirmara o negara su existencia. El objetivo de este trabajo fue encontrar dichas estructuras en las paredes ventriculares subendocárdicas de corazones humanos. Recolectamos 15 corazones conservados en formalina del Departamento de Anatomía de la Universidad de São Paulo y seccionamos las porciones apicales de los ventrículos derecho e izquierdo según el método utilizado por Goerttler. Utilizamos histología convencional (microscopía de luz-LM), microscopía electrónica de barrido (SEM) y un nuevo método de conservación llamado microplastinación (MP). En la pared anterior del ventrículo derecho en la región subendocárdica entre el tabique interventricular y la banda moderadora, encontramos varios haces de fibras fusiformes y helicoidales de histología similar a la FMV. Los haces medían entre 400 y 1150 µm de longitud y estaban separados de las fibras musculares adyacentes por una fina fibra de colágeno, actuando así como una "pseudocápsula". Algunas estructuras parecían estar vinculadas a la fibras de purkinje y parecían ser vasos linfáticos, sanguíneos y nerviosos. Llamamos a esas estructuras "corpúsculos cardíacos" (CC). La observación del CC previamente "desconocido" en este estudio inicial confirmó las descripciones anteriores y su descubrimiento puede contribuir a nuevas perspectivas en el estudio de la estructura y función del músculo cardíaco.

Prototyping and 3D printing of computed tomography images with an emphasis on soft tissues, especially muscles, for teaching human anatomy / Prototipado e impresión 3D de imágenes de tomografía computarizada con énfasis en los tejidos blandos, especialmente músculos, para la enseñanza de la anatomía humana

SUMMARY: The study on cadavers, although considered fundamental in the teaching of human anatomy, is limited in several universities, mainly due to the acquisition and manipulation of cadaveric material. Throughout history, several artificial anatomical models have been used to complement the real anatomical pieces. The present study offers a new alternative: the making of three-dimensional models from Computed Tomography (3D-CT) patient image acquisition. CT images from the USP University Hospital database were used. Patients underwent examinations for reasons other than the present study and were anonymized to maintain confidentiality. The CT slices obtained in thin cross-sections (approximately 1.0 mm thick) were converted into three-dimensional images by a technique named Volume Rendering for visualization of soft tissue and bone. The reconstructions were then converted to an STL (Standard Triangle Language) model and printed through two printers (LONGER LK4 Pro® and Sethi S3®), using PLA and ABS filaments. The 3D impressions of the thigh and leg muscles obtained better visual quality, being able to readily identify the local musculature. The images of the face, heart, and head bones, although easily identifiable, although seemed to present lower quality aesthetic results. This pilot study may be one of the first to perform 3D impressions of images from CT to visualize the musculature in Brazil and may become an additional tool for teaching.

El estudio en cadáveres, a pesar de considerarse un aspecto fundamental en la enseñanza de la anatomía humana, se encuentra limitado en varias universidades, principalmente por la adquisición y manipulación de material cadavérico. A lo largo de la historia se han utilizado varios modelos anatómicos artificiales para complementar las piezas anatómicas reales. El presente estudio ofrece una nueva alternativa: la elaboración de modelos tridimensionales a partir de la adquisición de imágenes de pacientes por Tomografía Computarizada (3D-CT). Se utilizaron imágenes de TC de la base de datos del Hospital Universitario de la USP. Los pacientes se sometieron a exámenes por razones distintas al presente estudio y fueron anonimizados para mantener la confidencialidad. Los cortes de TC obtenidos en secciones transversales delgadas (aproximadamente 1,0 mm de grosor) se convirtieron en imágenes tridimensionales mediante una técnica denominada Volume Rendering para la visualización de tejido blando y hueso. Luego, las reconstrucciones se convirtieron a un modelo STL (Standard Triangle Language) y se imprimieron a través de dos impresoras (LONGER LK4 Pro® y Sethi S3®), utilizando filamentos PLA y ABS. Se obtuvo una mejor calidad visual de las impresiones 3D de los músculos del muslo y la pierna, pudiendo identificar fácilmente la musculatura local. Las imágenes de la cara, el corazón y los huesos de la cabeza, aunque fácilmente identificables, parecían presentar resultados estéticos de menor calidad. Este estudio piloto puede ser uno de los primeros en realizar impresiones 3D de imágenes de TC para visualizar la musculatura y podría ser en una herramienta adicional para la enseñanza.

Age-related changes in urinary bladder intramural neurons.

A quantitative morphometric evaluation of the intramural plexus of the urinary bladder of adult and aged guinea-pigs was performed by histological analysis, scanning electron microscopy, and hystochemical methods, such as NADH-diaphorase and acetylcholinesterase (AChE). The round or oval shaped intramural neurons were revealed among the bundles of the smooth detrusor muscle in clusters containing a variable number of cells in the groups. In both adult control and aged animals, the ganglia were enveloped by a ganglionar capsule of connective tissue mainly composed of type I collagen fibers. The number of neurons NADH-diaphorase positives estimated in the intramural plexus was 1433+/-187.71 and 1107+/-120.67 in the adult control and aged groups, respectively. The perikaryon areas of the NADH-diaphorase neurons reactives ranged from 216.40 to 1809.30 microm(2) in adult control group and from 198.20 to 2096.25 microm(2) in aged group. The nuclear area showed an increase in aged animals. The number of AChE-positive neurons estimated in the intramural plexus was 3294.67+/-415 microm(2) in the adult control group and 1960.33+/-526 microm(2) in the aged group, showing a significant decrease in the latter group. This age-related morphological change in intramural neurons may contribute to changes in urinary bladder activities in the elderly.

Atrial natriuretic peptide (ANP)-granules in the guinea pig atrial and auricular cardiocytes: an immunocytochemical and ultrastructural morphometric comparative study.

The atrial natriuretic peptide (ANP) is a peptide hormone that is mainly produced in the cardiac atria, where it is stored within granules. It is known that the four regions of the atrial-auricular complex (two atria and two auricles) produce and store ANP in the granules. However, no report has been presented comparing the presence of ANP, and the number and diameter of atrial granules in the atria and auricles. ANP immunoreactivity was detected in cardiocytes from the four regions of the atrial-auricular complex. No differences were observed among the regions. The number of granules was greatest in the right atrium followed by the left atrium and left auricle and right auricle, in this order. The diameter of granules in the cardiocytes was significantly largest in the right atrium and reduced via the left auricle to the left atrium and right auricle. Both the number and diameter of the granules are larger in the right atrium in comparison with the other regions of the atrial-auricular complex, which leads to the supposition that this region is the one that most synthesizes and stores the ANP.

Protein Malnutrition Pre- and Postnatal and Nutritional Rehabilitation Modulates the Morphology of Muscle Fibers in Wistar Rats.

This study investigated the effects of pre- and postnatal conditions of protein deficiency followed to nutritional rehabilitation in the morphology of skeletal muscle. Twelve Wistar male rats were distributed in two groups: nourished (N), with normal protein diet and undernourished (U), with low protein diet. The respective diet was maintained until animals completed 21 days of life. After that, part of group U (n = 4) received normal protein diet, forming a third group, renourished group (R). Forty-two-day-old animals were euthanized and we performed histopathological and morphometric analysis of the soleus muscle. Analysis stained in hematoxylin and eosin (H&E) of the group N revealed polygonal and equidistant muscle fibers, with normal distribution in muscle fascicles. However, D group had rounded and disorganized fibers with different distances between them in muscle fascicles. R group presented muscle fibers with several formats, polygonal and rounded, and some muscle fascicles starting the reorganization process. In N group, analysis of the connective tissue showed predominance of type I collagen and a lower amount collagen type III, both well organized. Whereas U group had a predominance of disorganized type III collagen, in R group, there was return of type I collagen, but partially organized. Muscle fiber area of U (163.18 ± 52.55 µm2) and R (381.79 ± 26.62 µm2) groups was smaller than N (1229.2 µm2 ± 61.12 µm2). Muscle fibers density of groups U (3369 ± 1226 fibers/mm2) and R (1979 ± 28 fibers/mm2) was larger than N (830 ± 113 fibers/mm2). The nutritional rehabilitation in the present study showed an attempt of reorganization of the muscle tissue.

Protein malnutrition during gestation and early life decreases neuronal size in the medial prefrontal cortex of post-pubertal rats.

Retrospective studies in human populations indicate that protein deprivation during pregnancy and early life (early protein malnutrition, EPM) is associated with cognitive impairments, learning disabilities and may represent a risk factor for the late onset of some psychiatric disorders, fundamentally schizophrenia, a condition where the prefrontal cortex plays an important role. The purpose of this study was to analyze whether EPM affects structural aspects of the rat medial prefrontal cortex (mPFC), such as cortical volume, neuronal density and neuronal soma size, which seem altered in patients with schizophrenia. For this, a rat model of EPM (5% casein from conception to postnatal day 60) was adopted and the rat mPFC volume, total number of neurons and average neuronal volume were evaluated on postnatal day 60 (post-pubertal animals) by histo- and immunohistochemical techniques using unbiased stereological analysis. EPM did not alter the number of NeuN+ neurons in the rat mPFC. However, a very significant decrease in mPFC volume and average neuronal size was observed in malnourished rats. Although the present study does not establish causal relationships between malnutrition and schizophrenia, our results may indicate a similar structural phenomenon in these two situations.

Supplementation action with ascorbic acid in the morphology of the muscular layer and reactive acetylcholinesterase neurons of ileum of mdx mice.

The Duchenne Muscular Dystrophy (DMD) is a genetic disorder characterized by the absence of dystrophin protein, causing severe myopathy from increases of oxidative stress. Injuries of intestinal muscle can compromise the myenteric plexus. This study aimed to evaluate the disorders occurred in the muscular layer and in the acetylcholinesterase myenteric neurons (ACHE-r) of ileum of mdx mice, and the effects of supplementation with ascorbic acid (AA) in both components. 30 male mice C57BL/10, and 30 male mice C57BL/10Mdx were separated according to the age and treatment (n=10/group): 30-days-old control group (C30); 30-days-old dystrophic group (D30); 60-days-old control group (C60); 60-days-old dystrophic group (D60); 60-days-old control group supplemented with AA (CS60); and 60-days-old dystrophic group supplemented with AA (DS60). The animals were euthanized and the ileum was collected and processed. Semi-serial sections were stained by Masson's trichrome, and acetylcholinesterase histochemical technique in whole-mounts preparations to identify the myenteric neurons. The muscular layer thickness and the area of smooth muscle of ileum were lower in dystrophic groups, especially in D30 group. The DS60 group showed the muscular layer thickness similar to C60. The density of ACHE-r neurons of myenteric plexus of ileum was lower in D30 animals; however, it was similar in animals of 60-days-old without treatment (C60 and D60) and, higher in DS60. The cell body profile area of ACHE-r neurons was similar in C30-D30 and C60-D60; however, it was higher in DS60. DMD caused damage to the ileum's musculature and myenteric plexus, and the AA prevented the ACHE-r neuronal loss.

Combination of a high-fat diet with sweetened condensed milk exacerbates inflammation and insulin resistance induced by each separately in mice.

Obesogenic diets increase body weight and cause insulin resistance (IR), however, the association of these changes with the main macronutrient in the diet remains to be elucidated. Male C57BL/6 mice were fed with: control (CD), CD and sweetened condensed milk (HS), high-fat (HF), and HF and condensed milk (HSHF). After 2 months, increased body weight, glucose intolerance, adipocyte size and cholesterol levels were observed. As compared with CD, HS ingested the same amount of calories whereas HF and HSHF ingested less. HS had increased plasma AST activity and liver type I collagen. HF caused mild liver steatosis and hepatocellular damage. HF and HSHF increased LDL-cholesterol, hepatocyte and adipocyte hypertrophy, TNF-α by macrophages and decreased lipogenesis and adiponectin in adipose tissue (AT). HSHF exacerbated these effects, increasing IR, lipolysis, mRNA expression of F4/80 and leptin in AT, Tlr-4 in soleus muscle and IL-6, IL-1ß, VCAM-1, and ICAM-1 protein in AT. The three obesogenic diets induced obesity and metabolic dysfunction. HS was more proinflammatory than the HF and induced hepatic fibrosis. The HF was more detrimental in terms of insulin sensitivity, and it caused liver steatosis. The combination HSHF exacerbated the effects of each separately on insulin resistance and AT inflammatory state.

Incomplete petrosphenoidal foramen: morphological and morphometric analysis and the proposal of a classification study in Brazilian dry skulls / Foramen petrosfenoidal incompleto: análisis morfológico y morfométrico y propuesta de clasificación: estudio en cráneos secos brasileños

SUMMARY: The complete petrosphenoidal foramen, or canal, is an eventual and atavistic bony formation at the boundary between the posterior and middle cranial fossa, by occurrence of ossification of the superior petrosphenoidal ligament. This ligament ossification, which has important clinical and surgical significance, can be complete or incomplete, in variable degrees, and is associated with the passageway of neurovascular structures, such as the abducens nerve and the inferior petrosal sinus. This study, conducted with 175 dry skulls that belong to the University of São Paulo's collection (USP), São Paulo, Brazil, established criteria for a morphological classification of the incomplete petrosphenoid foramen in nine types. In addition, anatomical parameters were established for the morphometric determination of two diameters: the Oblique Diameter (ObDi) and the Maximum Transverse Diameter (MTD). Thus, of the 175 skulls, 146 (83.42 %) presented some of the incomplete forms of the petrosphenoid foramen, and 43 skulls (29.45 %), due to their conservation characteristics, were habilitated to the morphological study, in the classification and in the morphometry (the types I and II of our classification). The type II (incomplete foramen with bony projections of the petrosal tubercle, of the margin of the dorsum of the hypophyseal fossa or of the posterior clinoid process with a distance between them greater than 1mm) and type V (incomplete foramen with a bony projection only in one of the referential structures - posterior clinoid process) were the most common in this study (50 % of the 86 hemiskulls). Morphometry was attributed only to the types: I selar (incomplete foramen with bony projections from the petrosal tubercle and the margin of the dorsum of the hypophyseal fossa with a distance between them less than or equal to 1mm) and to the type II of this classification. The type I selar (9.3 % of the 43 skulls) resulted in an average of 3.25 mm of MTD and 4.63 mm, on average, of ObDi. The type II (25.58 % of the 43 skulls) showed, on average, 4.93 mm of MTD and 7.01 mm of ObDi.

RESUMEN: El foramen o canal petroesfenoidal completo es una formación ósea eventual y atávica en el límite entre las fosas craneal posterior y media, por osificación del ligamento petroesfenoidal superior. Esta osificación del ligamento, que tiene un importante significado clínico y quirúrgico, puede ser completa o incompleta, en grados variables, y está asociada al paso de estructuras neurovasculares, como el nervio abducente y el seno petroso inferior. Este estudio se realizó en 175 cráneos secos pertenecientes a la colección de la Universidad de São Paulo (USP), São Paulo, Brasil. Se establecieron criterios para una clasificación morfológica del foramen petrosfenoidal incompleto en nueve tipos. Además, se establecieron parámetros anatómicos para la determinación morfométrica de dos diámetros: el Diámetro Oblicuo (ObDi) y el Diámetro Transversal Máximo (MTD). Así, de los 175 cráneos, 146 (83,42 %) presentaron alguna de las formas incompletas del foramen petrosfenoidal, y 43 cráneos (29,45 %), por sus características de conservación, fueron habilitados para el estudio morfológico, en la clasificación y en la morfometría (los tipos I y II de nuestra clasificación). El Tipo II (foramen incompleto con proyecciones óseas del tubérculo petroso, del margen del dorso de la fosa hipofisaria o del proceso clinoides posterior con una distancia entre ellos mayor de 1 mm) y el Tipo V (foramen incompleto con proyección ósea solamente en una de las estructuras referenciales - proceso clinoides posterior) fueron los más comunes en este estudio (50 % de los 86 hemiscráneos). La morfometría se atribuyó únicamente al Tipo I selar (foramen incompleto con proyecciones óseas desde el tubérculo petroso y el margen del dorso de la fosa hipofisaria con una distancia entre ellos menor o igual a 1mm) y al Tipo II de esta clasificación. El Tipo I selar (9,3 % de los 43 cráneos) resultó en un promedio de 3,25 mm de MTD y 4,63 mm, en promedio, de ObDi. El Tipo II (25,58 % de los 43 cráneos) mostró, en promedio, 4,93 mm de MTD y 7,01 mm de ObDi.