From zero to ossified: Larval skeletal ontogeny of the Neotropical Cichlid fish Cichlasoma dimerus.

The identification of skeletal elements, the analysis of their developmental sequence, and the time of their appearance during larval development are essential to broaden the knowledge of each fish species and to recognize skeletal abnormalities that may affect further fish performance. Therefore, this study aimed to provide a general description of the development of the entire skeleton highlighting its variability in Cichlasoma dimerus. Larvae of C. dimersus were stained with alcian blue and alizarin red from hatching to 25 days posthatching. Skeletogenesis began with the endoskeletal disk and some cartilage structures from the caudal fin and the splachnocranium, while the first bony structure observed was the cleithrum. When larvae reached the free-swimming and exogenous feeding stage, mostly bones from the jaws, the branchial arches, and the opercle series evidenced some degree of ossification, suggesting that the ossification sequence of C. dimerus adjusts to physiological demands such as feeding and ventilation. The caudal region was the most variable regarding meristic counts and evidenced higher incidence of bone deformities. In conclusion, this work provides an overview of C. dimerus skeletogenesis and lays the groundwork for further studies on diverse topics, like developmental plasticity, rearing conditions, or phylogenetic relationships.

Effects of atracurium besylate on corneal endothelium of chickens: in vitro study / Avaliação dos efeitos do besilato de atracúrio no endotélio corneano de galinhas: estudo in vitro

The aim of this study was to investigate the acute effects of atracurium besylate on cellular damage in corneal endothelium of chickens. Twenty healthy chicken eyes were assigned to the following groups: Group 1 (G1), experimental group (n=10); and Group 2 (G2), control (n=10). Excised corneoscleral buttons were immediately placed on glass microscopy slides with endothelial region faced up. Corneal endothelium of eyes in G1 were covered with AB (0.2mL, 10mg/mL) for 3 min and then rinsed with balanced salt solution (BSS), while the corneal endothelium of eyes in G2 were covered with BBS for 3 min. Corneas from both groups were stained with alizarin red/trypan blue and visualized by light microscopy. Ten random photographs were taken from each cornea. The area of cellular damage was measured by software in all samples and cell loss of each group was averaged and compared. Endothelial area of denudation and Descemet's membrane exposure were higher in G1 than G2. In conclusion, atracurium besylate induced an acute damage on corneal endothelium of chickens.(AU)'

Objetivou-se avaliar os efeitos agudos do besilato de atracúrio sobre o endotélio corneano de galinhas. Vinte olhos saudáveis de galinhas foram aleatoriamente separados em dois grupos com 10 olhos cada, sendo G1 o grupo controle e G2 o grupo tratamento. Imediatamente após a excisão dos botões corneoesclerais estes foram colocados em lâminas de microscopia de vidro com o lado endotelial voltado para cima. No Grupo 1, o endotélio corneano foi recoberto com 0,2ml de besilato de atracúrio (10mg/ml) durante 3 minutos e depois lavado com solução salina balanceada. No Grupo 2, o endotélio corneano foi recoberto apenas com solução salina balanceada durante 3 min. As córneas de ambos os grupos foram coradas com vermelho de alizarina e azul de tripano e visualizadas com microscópio óptico. Foram obtidas dez fotografias aleatórias de cada amostra. As imagens foram analisadas e com auxílio de um software as áreas com ausência de células endoteliais calculadas. A perda celular endotelial foi significativamente maior no grupo tratamento comparativamente ao grupo controle. Com base nos resultados apresentados foi possível concluir que o besilato de atracúrio induziu dano agudo nas células do endotélio da córnea de galinhas.(AU)

Effects of atracurium besylate on corneal endothelium of chickens: in vitro study / Avaliação dos efeitos do besilato de atracúrio no endotélio corneano de galinhas: estudo in vitro

The aim of this study was to investigate the acute effects of atracurium besylate on cellular damage in corneal endothelium of chickens. Twenty healthy chicken eyes were assigned to the following groups: Group 1 (G1), experimental group (n=10); and Group 2 (G2), control (n=10). Excised corneoscleral buttons were immediately placed on glass microscopy slides with endothelial region faced up. Corneal endothelium of eyes in G1 were covered with AB (0.2mL, 10mg/mL) for 3 min and then rinsed with balanced salt solution (BSS), while the corneal endothelium of eyes in G2 were covered with BBS for 3 min. Corneas from both groups were stained with alizarin red/trypan blue and visualized by light microscopy. Ten random photographs were taken from each cornea. The area of cellular damage was measured by software in all samples and cell loss of each group was averaged and compared. Endothelial area of denudation and Descemet's membrane exposure were higher in G1 than G2. In conclusion, atracurium besylate induced an acute damage on corneal endothelium of chickens.(AU)'

Objetivou-se avaliar os efeitos agudos do besilato de atracúrio sobre o endotélio corneano de galinhas. Vinte olhos saudáveis de galinhas foram aleatoriamente separados em dois grupos com 10 olhos cada, sendo G1 o grupo controle e G2 o grupo tratamento. Imediatamente após a excisão dos botões corneoesclerais estes foram colocados em lâminas de microscopia de vidro com o lado endotelial voltado para cima. No Grupo 1, o endotélio corneano foi recoberto com 0,2ml de besilato de atracúrio (10mg/ml) durante 3 minutos e depois lavado com solução salina balanceada. No Grupo 2, o endotélio corneano foi recoberto apenas com solução salina balanceada durante 3 min. As córneas de ambos os grupos foram coradas com vermelho de alizarina e azul de tripano e visualizadas com microscópio óptico. Foram obtidas dez fotografias aleatórias de cada amostra. As imagens foram analisadas e com auxílio de um software as áreas com ausência de células endoteliais calculadas. A perda celular endotelial foi significativamente maior no grupo tratamento comparativamente ao grupo controle. Com base nos resultados apresentados foi possível concluir que o besilato de atracúrio induziu dano agudo nas células do endotélio da córnea de galinhas.(AU)

Morphology of endothelial cells from different regions of the swine cornea

Background: The corneal endothelium is a monolayer of polygonal cells which constitute the last layer of the cornea. The integrity of this layer is critical to cornea transparency. The characterization of normal corneal endothelial morphology is important not only to clinical evaluation but also to selection of areas of the cornea with better quality to be employed as donor tissue. The aim of the present study was to evaluate the morphology of endothelial cells from different regions of the swine cornea after alizarin red staining using optical microscopy.Materials, Methods & Results: Twenty-four healthy eyes from 12 swine Large White breed, with 14-monthold, males or females obtained from a licensed Brazilian commercial slaughterhouse were studied. Immediately after humane slaughter, the eyes were enucleated and submitted to ophthalmic examination. Eyes with signs of diseases of the anterior segment were excluded. The cornea, with 3 mm of the sclera, was removed and placed on a glass microscope slide with the endothelial side up. Four radial incisions were made in the periphery of the cornea to better accommodate the cornea on the microscope slide. Alizarin red was diluted in isotonic solution (0.2 g/100 mL) and the pH was adjusted to 4.2 with hydrochloric acid. Three drops of alizarin red were placed on the corneal endothelium. After 90 s, the dye was removed from the cornea with balanced saline solution. The corneal endothelium was examined and photographed using an optical microscope. All evaluations were performed by the same investigator. Photomicrographs were taken of central, superior, inferior, nasal and temporal corneal areas. Parameter studied included endothelial cell morphology. For the statistical analysis, was employed the ANOVA variance test (repeated measures). Differences were considered statistically significant at P < 0.05.[...](AU)

Morphology of endothelial cells from different regions of the swine cornea

Background: The corneal endothelium is a monolayer of polygonal cells which constitute the last layer of the cornea. The integrity of this layer is critical to cornea transparency. The characterization of normal corneal endothelial morphology is important not only to clinical evaluation but also to selection of areas of the cornea with better quality to be employed as donor tissue. The aim of the present study was to evaluate the morphology of endothelial cells from different regions of the swine cornea after alizarin red staining using optical microscopy.Materials, Methods & Results: Twenty-four healthy eyes from 12 swine Large White breed, with 14-monthold, males or females obtained from a licensed Brazilian commercial slaughterhouse were studied. Immediately after humane slaughter, the eyes were enucleated and submitted to ophthalmic examination. Eyes with signs of diseases of the anterior segment were excluded. The cornea, with 3 mm of the sclera, was removed and placed on a glass microscope slide with the endothelial side up. Four radial incisions were made in the periphery of the cornea to better accommodate the cornea on the microscope slide. Alizarin red was diluted in isotonic solution (0.2 g/100 mL) and the pH was adjusted to 4.2 with hydrochloric acid. Three drops of alizarin red were placed on the corneal endothelium. After 90 s, the dye was removed from the cornea with balanced saline solution. The corneal endothelium was examined and photographed using an optical microscope. All evaluations were performed by the same investigator. Photomicrographs were taken of central, superior, inferior, nasal and temporal corneal areas. Parameter studied included endothelial cell morphology. For the statistical analysis, was employed the ANOVA variance test (repeated measures). Differences were considered statistically significant at P < 0.05.[...]

Evaluation of equine corneal endothelium after exposure to 0.5% indocyanine green - in vitro study / Avaliação do endotélio corneano de equinos após exposição à indocianina verde 0.5% - estudo in vitro

The purpose of the study was to investigate whether indocyanine green (ICG) dye damages the corneal endothelium of horses. Twenty-four corneas of 12 healthy equines, males or females, of different ages were used in this study. Only eyes with no ocular findings were used. Randomly, one eye was included in the treatment group and one in the control group. The eyes of the treatment group were exposed for 1 minute to dye ICG 0.5%. After that the endothelium of all eyes was stained with trypan blue and alizarin red S and analyzed and photographed under an optical microscope. Areas with damaged endothelial cells were manually measured and quantified using software for morphometric analysis and expressed as a percentage of cell damage. In all eyes examined areas of cell damage were observed in both corneas of the control group and the treatment group. The mean endothelial damage was 0.8 ± 0.37% in the treatment group and 0.97 ± 0.39% in the control. The Qui-square test stated that treatment and control group were not different. The ICG 0.5% did not cause acute damage to equine corneal endothelium.

O objetivo do estudo foi investigar se indocianina verde (ICG) induz dano nas células do endotélio da cَrnea de equinos. Vinte e quatro cَrneas de 12 equinos saudلveis, machos ou fêmeas, de diferentes idades foram estudadas. Somente olhos hيgidos foram utilizados. Aleatoriamente, um olho foi incluيdo no grupo controle e outro no grupo tratamento. Os olhos do grupo tratamento foram expostos durante um minuto à indocianina verde a 0,5%. Posteriormente, o endotélio da cَrnea foi corado com azul de tripano e vermelho de alizarina, analisado e fotografado usando microscَpio َptico. As لreas com células endoteliais danificadas foram aferidas e quantificadas utilizando um software para anلlise morfométrica. Os valores encontrados foram expressos como percentual de perda celular. Em todos os olhos examinados foram observadas لreas de dano celular, tanto no grupo controle quanto no grupo tratamento. A perda celular endotelial média foi de 0,8±0,37% no grupo tratamento e 0,97 ± 0,39% no grupo controle. O teste Qui-quadrado confirmou que os grupos tratamento e controle nمo diferiram. Foi possيvel concluir que a ICG 0,5% nمo causou dano agudo nas células do endotélio da cَrnea de equinos.

Evaluation of equine corneal endothelium after exposure to 0.5% indocyanine green - in vitro study / Avaliação do endotélio corneano de equinos após exposição à indocianina verde 0.5% - estudo in vitro

The purpose of the study was to investigate whether indocyanine green (ICG) dye damages the corneal endothelium of horses. Twenty-four corneas of 12 healthy equines, males or females, of different ages were used in this study. Only eyes with no ocular findings were used. Randomly, one eye was included in the treatment group and one in the control group. The eyes of the treatment group were exposed for 1 minute to dye ICG 0.5%. After that the endothelium of all eyes was stained with trypan blue and alizarin red S and analyzed and photographed under an optical microscope. Areas with damaged endothelial cells were manually measured and quantified using software for morphometric analysis and expressed as a percentage of cell damage. In all eyes examined areas of cell damage were observed in both corneas of the control group and the treatment group. The mean endothelial damage was 0.8 ± 0.37% in the treatment group and 0.97 ± 0.39% in the control. The Qui-square test stated that treatment and control group were not different. The ICG 0.5% did not cause acute damage to equine corneal endothelium.(AU)

O objetivo do estudo foi investigar se indocianina verde (ICG) induz dano nas células do endotélio da cَrnea de equinos. Vinte e quatro cَrneas de 12 equinos saudلveis, machos ou fêmeas, de diferentes idades foram estudadas. Somente olhos hيgidos foram utilizados. Aleatoriamente, um olho foi incluيdo no grupo controle e outro no grupo tratamento. Os olhos do grupo tratamento foram expostos durante um minuto à indocianina verde a 0,5%. Posteriormente, o endotélio da cَrnea foi corado com azul de tripano e vermelho de alizarina, analisado e fotografado usando microscَpio َptico. As لreas com células endoteliais danificadas foram aferidas e quantificadas utilizando um software para anلlise morfométrica. Os valores encontrados foram expressos como percentual de perda celular. Em todos os olhos examinados foram observadas لreas de dano celular, tanto no grupo controle quanto no grupo tratamento. A perda celular endotelial média foi de 0,8±0,37% no grupo tratamento e 0,97 ± 0,39% no grupo controle. O teste Qui-quadrado confirmou que os grupos tratamento e controle nمo diferiram. Foi possيvel concluir que a ICG 0,5% nمo causou dano agudo nas células do endotélio da cَrnea de equinos.(AU)

Morphology of endothelial cells from different regions of the cornea of dogs / Morfologia das células endoteliais de diferentes regiões da córnea de cães

ABSTRACT: The aim of this study was to evaluate the morphology of endothelial cells from different areas of the cornea of dogs. Twenty healthy eyes from 10 dogs, females or males, of different ages were studied. Corneal endothelium morphology of superior, inferior, central, nasal and temporal areas was assessed by 0.2% alizarin red staining using an optic microscope. One hundred endothelial cells from each corneal area were analyzed. In all areas of the cornea studied were found endothelial cells with four sides, five sides, six sides and seven sides. There was no significant difference regarding endothelial cell morphology in all corneal regions evaluated. Thus, the morphology of the central cornea area represents the entire endothelial mosaic and may be applied to peripheral areas. Therefore, analysis of the central area is sufficient to estimate the shape of endothelial cells of peripheral areas of healthy dog corneas.

RESUMO: Objetivou-se avaliar a morfologia das células endoteliais de diferentes regiões da córnea de cães. Vinte olhos saudáveis de 10 cães, fêmeas ou machos, de diferentes idades foram estudados. A morfologia do endotélio corneano das regiões superior, inferior, central, nasal e temporal foi avaliada pela coloração vermelho de alizarina 0,2% com microscópio óptico. Foram analisadas 100 células endoteliais de cada região da córnea. Em todas as regiões da córnea estudadas foram encontradas células endoteliais com quatro lados, cinco lados, seis lados e sete lados. Não houve diferença significativa em relação à morfologia de células endoteliais da córnea em todos as regiões estudadas. Assim, a morfologia da região central da córnea representa todo o mosaico endotelial e pode ser aplicada em áreas periféricas. Portanto, a análise da área central é suficiente para estimar a forma das células endoteliais das áreas periféricas de córneas de cães saudáveis.

Morphology of endothelial cells from different regions of the cornea of dogs / Morfologia das células endoteliais de diferentes regiões da córnea de cães

The aim of this study was to evaluate the morphology of endothelial cells from different areas of the cornea of dogs. Twenty healthy eyes from 10 dogs, females or males, of different ages were studied. Corneal endothelium morphology of superior, inferior, central, nasal and temporal areas was assessed by 0.2% alizarin red staining using an optic microscope. One hundred endothelial cells from each corneal area were analyzed. In all areas of the cornea studied were found endothelial cells with four sides, five sides, six sides and seven sides. There was no significant difference regarding endothelial cell morphology in all corneal regions evaluated. Thus, the morphology of the central cornea area represents the entire endothelial mosaic and may be applied to peripheral areas. Therefore, analysis of the central area is sufficient to estimate the shape of endothelial cells of peripheral areas of healthy dog corneas.(AU)

Objetivou-se avaliar a morfologia das células endoteliais de diferentes regiões da córnea de cães. Vinte olhos saudáveis de 10 cães, fêmeas ou machos, de diferentes idades foram estudados. A morfologia do endotélio corneano das regiões superior, inferior, central, nasal e temporal foi avaliada pela coloração vermelho de alizarina 0,2% com microscópio óptico. Foram analisadas 100 células endoteliais de cada região da córnea. Em todas as regiões da córnea estudadas foram encontradas células endoteliais com quatro lados, cinco lados, seis lados e sete lados. Não houve diferença significativa em relação à morfologia de células endoteliais da córnea em todos as regiões estudadas. Assim, a morfologia da região central da córnea representa todo o mosaico endotelial e pode ser aplicada em áreas periféricas. Portanto, a análise da área central é suficiente para estimar a forma das células endoteliais das áreas periféricas de córneas de cães saudáveis.(AU)

Mast Cell Mediators Inhibit Osteoblastic Differentiation and Extracellular Matrix Mineralization.

Mast cells are multifunctional immune cells that participate in many important processes such as defense against pathogens, allergic reactions, and tissue repair. These cells perform their functions through the release of a wide variety of mediators. This release occurs mainly through cross-linking IgE (immunoglobulin E) bound to high affinity IgE receptors by multivalent antigens. The abundance of mast cells in connective tissue, surrounding blood vessels, and their involvement in the early stages of bone repair support the possibility of physiological and pathological interactions between mast cells and osteoblasts. However, the participation of mast cell mediators in osteogenesis is not fully understood. Therefore, the objective of this work was to investigate the role of mast cell mediators in the acquisition of the osteogenic phenotype in vitro. The results show that pooled mast cell mediators can affect proliferation, morphology, and cytoskeleton of osteoblastic cells, and impair the activity and expression of alkaline phosphatase as well as the expression of bone sialoprotein. Also, mast cell mediators inhibit the expression of mRNA for those proteins and inhibit the formation and maturation of calcium nodules and consequently inhibit mineralization. Therefore, mast cell mediators can modulate osteogenesis and are potential therapeutic targets for treatments of bone disorders.

Morphology of endothelial cells from different regions of the equine cornea / Morfologia das células do endotélio de diferentes regiões da córnea de equinos

ABSTRACT: The objective of this study was to evaluate the morphology of different regions of the equine cornea using optical microscopy. Both healthy eyes of eight horses, male or female, of different ages were evaluated. Corneas were stained with alizarin red vital dye and subsequently examined and photographed using optical microscopy. Corneal endothelial morphology of central, superior, inferior, temporal and nasal areas was assessed. One hundred endothelial cells from each corneal area were analyzed. The shape of the corneal endothelial cells of each corneal region was analyzed. Statistical data analysis was conducted using the Student's t test. Values of P<0.01 were considered significant. Regarding morphological analysis, no statistically significant differences were reported between the equine corneal regions analyzed. The present research suggested that there are no morphological differences between regions of the equine cornea. The values obtained in any region of the equine cornea can be extrapolated to other regions of the cornea and are representative of the cell morphology present in all regions of the cornea.

RESUMO: O objetivo deste estudo foi o de avaliar a morfologia das diferentes regiões da córnea equina usando microscopia óptica. Foram avaliados ambos os olhos saudáveis ​​de oito equinos, machos ou fêmeas, de diferentes idades. As córneas foram coradas com corante vital vermelho de alizarina, examinadas com microscópio óptico e fotografadas. A morfologia endotélio corneano de áreas centrais, superior, inferior, temporal e nasal foi avaliada. Foram analisadas ​​células endoteliais da córnea de cada área. A forma das células endoteliais da córnea de cada região da córnea foi analisada. Análise estatística dos dados foi realizada por meio do teste t. Valores de P<0.01 foram considerados significativos. Em relação à análise morfológica estatisticamente significativa, não foram encontradas diferenças entre as regiões da córnea equina analisadas. O presente trabalho sugere que não houve diferença entre a morfologia das regiões da córnea de equino. Os valores obtidos em qualquer região da córnea equina podem ser extrapolados para outras regiões da córnea e são representativos da morfologia celular em todas as regiões da córnea.

Morphology of endothelial cells from different regions of the equine cornea / Morfologia das células do endotélio de diferentes regiões da córnea de equinos

The objective of this study was to evaluate the morphology of different regions of the equine cornea using optical microscopy. Both healthy eyes of eight horses, male or female, of different ages were evaluated. Corneas were stained with alizarin red vital dye and subsequently examined and photographed using optical microscopy. Corneal endothelial morphology of central, superior, inferior, temporal and nasal areas was assessed. One hundred endothelial cells from each corneal area were analyzed. The shape of the corneal endothelial cells of each corneal region was analyzed. Statistical data analysis was conducted using the Student's t test. Values of P<0.01 were considered significant. Regarding morphological analysis, no statistically significant differences were reported between the equine corneal regions analyzed. The present research suggested that there are no morphological differences between regions of the equine cornea. The values obtained in any region of the equine cornea can be extrapolated to other regions of the cornea and are representative of the cell morphology present in all regions of the cornea.(AU)

O objetivo deste estudo foi o de avaliar a morfologia das diferentes regiões da córnea equina usando microscopia óptica. Foram avaliados ambos os olhos saudáveis de oito equinos, machos ou fêmeas, de diferentes idades. As córneas foram coradas com corante vital vermelho de alizarina, examinadas com microscópio óptico e fotografadas. A morfologia endotélio corneano de áreas centrais, superior, inferior, temporal e nasal foi avaliada. Foram analisadas células endoteliais da córnea de cada área. A forma das células endoteliais da córnea de cada região da córnea foi analisada. Análise estatística dos dados foi realizada por meio do teste t. Valores de P<0.01 foram considerados significativos. Em relação à análise morfológica estatisticamente significativa, não foram encontradas diferenças entre as regiões da córnea equina analisadas. O presente trabalho sugere que não houve diferença entre a morfologia das regiões da córnea de equino. Os valores obtidos em qualquer região da córnea equina podem ser extrapolados para outras regiões da córnea e são representativos da morfologia celular em todas as regiões da córnea.(AU)

Validation of daily microincrement deposition in otoliths of juvenile and adult Peruvian anchovy Engraulis ringens.

Wild adult specimens of the Peruvian anchovy Engraulis ringens were captured and reared to validate the daily periodicity of otolith microincrement formation. The postcapture stress generated spontaneous spawning, making it possible to conduct a rearing trial on larvae first in an artificial nutrient-enriched system (ANES) for 52 days followed by an artificial feeding regime in a culture tank until day 115 post-hatch. Microincrements of the sagittal otoliths of sacrificed juveniles [mean ± S.D. total length (LT ) = 5·13 ± 0·37 cm, range 5-6 cm; c.v. = 7·5%] showed very distinct light and dark zones. The slope of the relationship between the total number of increments after the hatch check and days elapsed after hatching was not significantly different from 1. The transfer from ANES to the artificial feeding regime induced a mark in the sagittal otoliths. The number of microincrements after this induced mark coincided with the number of days elapsed after the transfer date. In parallel experiments, adult E. ringens (mean ± S.D. LT = 14·92 ± 0·55 cm, range 13-16 cm) were exposed to one of two fluorescent marking immersion treatments with either alizarin red S (ARS; 25 mg l(-1) per 6 h) or oxytetracycline hydrochloride (OTC; 200 mg l(-1) per 10 h). The microincrements between fluorescent bands were distinct, ranging from 0·89 to 2·75 µm (mean ± S.D. =1·43 ± 0·28 µm; c.v. = 32%) and from 0·71 to 2·89 µm (1·53 ± 0·27 µm; c.v. = 35%) for ARS and OTC, respectively. The relationship between the number of microincrements between marks and the number of elapsed days for ARS and OCT treatments indicated that there was a significant correspondence between the number of increases observed and the number of days. Hence, daily microincrements of otoliths of E. ringens are likely to be formed in juveniles and adults under natural conditions.

Uncoupling of osteoblast-osteoclast regulation in a chemical murine model of Gaucher disease.

Gaucher disease (GD) is caused by mutations in the GBA gene that confer a deficient level of activity of glucocerebrosidase (GCase). This deficiency leads to accumulation of the glycolipid glucocerebroside in the lysosomes of cells of monocyte/macrophage system. Type I GD is the mildest form and is characterized by the absence of neuronopathic affection. Bone compromise in Gaucher disease patients is the most disabling aspect of the disease. However, pathophysiological aspects of skeletal alterations are still poorly understood. The homeostasis of bone tissue is maintained by the balanced processes of bone resorption by osteoclasts and formation by osteoblasts. We decided to test whether bone resorption and/or bone formation could be altered by the use of a chemical in vitro murine model of Gaucher disease. We used two sources of cells from monocyte/macrophages lineage isolated from normal mice, splenocytes (S) and peritoneal macrophages (PM), and were exposed to CBE, the inhibitor of GCase (S-CBE and PM-CBE, respectively). Addition of both conditioned media (CM) from S-CBE and PM-CBE induced the differentiation of osteoclasts precursors from bone marrow to mature and functional osteoclasts. TNF-α could be one of the factors responsible for this effect. On the other hand, addition of CM to an osteoblast cell culture resulted in a reduction in expression of alkaline phosphatase and mineralization process. In conclusion, these results suggest implication of changes in both bone formation and bone resorption and are consistent with the idea that both sides of the homeostatic balance are affected in GD.

Evaluation of marking efficiency of different alizarin red S concentrations on body fish structures in Oreochromis niloticus (Perciformes: Cichlidae) juveniles

The use of alizarin red S (ARS) marked tilapias could provide valuable fisheries management information to evaluate fish stocking events and may facilitate aquaculture management practices. As a new technique in fishes, the aim of this study was to compare and evaluate the chemical marks produced in tilapia juveniles by ARS through two treatments: 1) 12 hours of immersion and 2) immersion after osmotic induction. This was analyzed at three concentrations: 50, 75 and 100mg/l, and in three structures: otoliths, fish scales and caudal fin rays of Oreochromis niloticus juveniles. After three culture months 80% of specimens were analyzed and significant differences (p<0.05) in mark intensity were detected between treatments for otoliths and fin rays, but not for fish scales. Significant differences between concentrations were found for the 12h immersion treatment, while no significant differences were detected with osmotic induction. Our results showed that marks appeared at all concentrations, and none of the concentrations produced weak marks. Osmotic induction had a greater mortality than the 12h immersion procedure. After eight culture months the rest of the specimens were analyzed and the mark permanence was observed in all cases. According to the present results we recommend the marking process of 12h immersion treatment at 100mg/L concentration.

El uso de alizarina roja S (ARS) para marcar tilapias podría proporcionar información valiosa para el manejo de su pesquería. Para evaluar pesquerías acuaculturales manejadas con siembras o repoblamientos de peces se comparó y evaluó la marca producida por la alizarina roja S, empleando dos tratamientos: 1) Inmersión en ARS durante 12h; e 2) Inmersión en ARS después de un choque osmótico. El análisis se realizó a tres concentraciones: 50, 75 y 100mg/l y en tres estructuras: otolitos, escamas y radios de la aleta caudal de Oreochromis niloticus. Ochenta por ciento de los ejemplares fueron cultivados durante tres meses y analizados posteriormente. Los resultados mostraron diferencias entre las concentraciones de la marca para el tratamiento de 12h de inmersión mientras que no hubo diferencias entre las concentraciones para el tratamiento con inducción osmótica. Se encontraron diferencias en la intensidad de la marca entre los tratamientos para otolitos y radios de las aletas pero para las escamas no hubo diferencias significativas. Todas las concentraciones produjeron marcas (desde débiles a intensas), sin embargo la concentración de 100mg/l no produjo marcas débiles. El tratamiento por inducción osmótica presentó mayores niveles de mortalidad. Después de ocho meses de cultivo el resto de los ejemplares fueron analizados y se observó la permanencia de las marcas en todos los casos. En vista de lo anterior, para los propósitos de marcaje se recomienda el uso del tratamiento de inmersión por 12h y una concentración de 100mg/l.

Prenatal stress caused by movement restriction induces changes in the appendicular osseous development of CF-1 mice progeny / El estrés prenatal causado por restricción de movimiento induce cambios en el desarrollo óseo apendicular de la progenie de ratones CF-1

Prenatal stress has been associated with alterations in weight and body size, as well as disturbances in the process of developing skeletal ossification, occurring during childbirth and the early stages of life. However, the effect evidence of prenatal stress on bone grow thand development during the gestation period has been low; therefore, it is unknown whether these alterations are associated with potential for growth disorders Because of this, the study aims to determine the short-term effects of prenatal stress on the CF-1mouse bone structure growth in your date of birth. The female mice were divided randomly in two groups: controlled (n=2) and stressed (n=3). The latter was put under stress by means of movement restriction during the last week of gestation. Second, an evaluation of their gestational development was made, obtaining measurements of their weight. Finally, diaphanization with KOH and staining with Alizarin red was used to measure the length of their appendicular bones and their flat pelvic bones, of 53 P0 mice (25 control; 28 stressed during gestation). The stressed mice's body weight (p=0.02) and the length of their appendicular bones (radii, p=0.0011; ulnae, p= 0.0001; humeri, p=0.0001; femorae, p=0.0006; tibiae, p=0.0015) were affected significantly in contrast with the controlled group. On the other hand,there were nosignificant differences inmaternal bodyweightand length ofthe mice pelvic bones (isquium, ilium; p>0.05). The prenatal stress by means of movement restriction alters the osseous appendicular morphology of the CF-1 mouse evaluated at birth...

El estrés prenatal se ha asociado con alteraciones en el peso y el tamaño corporal, así como trastornos en el proceso osificación en el esqueleto en desarrollo, que se producen durante el parto y las primeras etapas de vida. Sin embargo, la evidencia de los efectos del estrés prenatal sobre el crecimiento de los huesos y el desarrollo durante el período de gestación ha sido baja, desconociéndose si estas alteraciones están relacionadas con trastornos del crecimiento. El objetivo fue determinar los efectos a corto plazo del estrés prenatal en la estructura ósea del ratón CF-1 en el día de nacimiento. Las hembras gestantes fueron divididas aleatoriamente en dos grupos: control (n=2) y estresado (n=3). Este último fue puesto bajo estrés por restricción de movimiento durante la última semana de gestación. Evaluando el peso corporal en la progenie al nacer (grupo control n=25; grupo estresado n=28), para posteriormente diafanizar mediante KOH y teñir con alizarina roja, midiendo la longitud de huesos largos apendiculares y huesos planos de pelvis en el plano coronal. El peso de los ratones estresados (p = 0,02) y la longitud de los huesos apendiculares fueron significativamente menores en comparación con el grupo control (radio, p = 0,0011; ulna, p = 0,0001; húmero, p = 0,0001; fémur, p = 0,0006; tibia, p = 0,0015). Por otro lado, no hubo diferencias significativas en el peso corporal de la madre y la longitud de los huesos pélvicos de las crías (isquion, íleon, p> 0,05). El estrés prenatal por restricción de movimiento altera la morfología ósea apendicular del ratón CF-1, evaluados al nacimiento...

Petroleum ether extract of Cissus quadrangularis (LINN) stimulates the growth of fetal bone during intra uterine developmental period: a morphometric analysis

OBJECTIVE: The aim of the present study was to analyze the effect Cissus quadrangularis plant petroleum ether extract on the development of long bones during the intra-uterine developmental stage in rats. METHODS: Pregnant rats (n=12) were randomly assigned into either a control group (n=6) or a Cissus quadrangularis treatment (n=6) group. Pregnant rats in the Cissus quadrangularis group were treated with Cissus quadrangularis petroleum ether extract at a dose of 500 mg/kg body weight from gestation day 9 until delivery. The animals in the control group received an equal volume of saline. Newborn pups were collected from both groups for alizarin red S - alcian blue staining to differentiate ossified and unossified cartilage. The ossified cartilage (bone) was morphometrically analyzed using Scion image software. RESULTS: Morphometric analysis revealed that the percentage of the total length of ossified cartilage (bone) in pups born to treated dams was significantly higher (P<0.001- -0.0001) than that of the control group. CONCLUSION: The results of the present study suggest that maternal administration of Cissus quadrangularis petroleum ether extract during pregnancy can stimulate the development of fetal bone growth during the intra-uterine developmental period.