RESUMEN
Background: Topically administered 2% dorzolamide is among the most commonly used agents to lower IOP. As a complication of glaucoma, blind patients may develop corneal ulcers secondary to trauma. Nonetheless, in patients with a hypertensive or glaucomatous eye, in which the cornea has also been ulcerated, medical hypotensive therapy should not be discontinued. Therefore, the present study aimed to determine whether the instillation of a benzalkonium chloride (BAK)-preserved 2% dorzolamide alters corneal wound healing time and the levels of matrix metalloproteinases (MMP-9) in the tears of cats with experimentally induced corneal ulcers. Materials, Methods & Results: Sixteen cats (8/group) were randomly assigned to receive 40 µL of 2% dorzolamide (TG) or saline (CG) 3 times daily until corneal re-epithelialization. Experimental keratectomies were performed under general and topical anesthesia using an operating microscope. For this purpose, a millimitred trephine was calibrated and used to create a temporal paraxial corneal ulcer with a diameter of 6 mm and a depth of 200 µm. After corneal wounding, the ulcerated area, the healing time, blepharospasm, conjunctival hyperemia, and aqueous flare were compared between groups. Tears were collected at baseline and 24 and 48 h after keratectomy, and the total MMP-9 was quantified by ELISA. Data were assessed statistically using unpaired Student's t test, one-way, and two-way ANOVA followed by a Bonferroni post hoc test. Statistical significance was set at P < 0.05 for all analyses. The average time to achieve corneal wound healing did not differ between groups (P = 0.36) and was 65.50 ± 3.62 h in the CG and 71.00 ± 4.58 h in the TG. Twenty-four h after keratectomy, the ulcerated area in the CG was 3.34 mm2 larger than that observed in the TG (P = 0.04); the rest of the comparisons did not reach statistical significance at any time point between groups (P > 0.05). Higher blepharospasm scores were observed in cats of TG (P = 0.04). When compared with baseline of both groups, the levels of MMP-9 increased significantly at 24 and 48 h post-keratectomy (P < 0.001), but differences between groups were not observed at 24 and 48 h post-keratectomy (P > 0.05). Discussion: In cats, 9 mm axial corneal ulcers created by superficial debridement re-epithelize approximately 48 h postwounding. In the present study, re-epithelialization post keratectomy occurred within an average time of 68.25 h in most cats and in a delayed manner in one cat of the TG after 96 h. In the current study, the lesions in both groups healed without corneal scarring, pigmentation, or vascularization. Although BAC was present in all topical medications used in the present study, the authors attribute the higher scores of blepharospasm in the TG to the rheological characteristics and the pH of the dorzolamide ophthalmic solution. Indeed, the pH value of dorzolamide (5.58) may cause signs of irritation, as the tear film has an approximate pH of 7.6. Previous studies showed that ulcerated corneas presented significantly higher levels of MMP-9 in tears at the early stages (8 to 36 h) post-wounding. In the current study, the levels of this enzyme after wounding did not change significantly in the tears of cats treated with 2% dorzolamide when compared to the eyes in the control group. This study showed that the instillation of a BAC-preserved 2% dorzolamide ophthalmic solution did not impair the corneal wound healing time or the early expression of MMP-9 in the tears of cats with experimentally induced corneal ulcers. However, our results warrant further investigation in patients with ocular hypertension or glaucoma presenting concomitant naturally occurring corneal ulcers to certify our findings.
Asunto(s)
Animales , Gatos , Úlcera de la Córnea/veterinaria , Glaucoma/veterinaria , Metaloproteinasa 9 de la Matriz/análisis , Epitelio/fisiología , Compuestos de Benzalconio/uso terapéutico , Inhibidores de Anhidrasa CarbónicaRESUMEN
Cystic fibrosis (CF) is an autosomal recessive disorder, caused by diverse genetic variants for the CF transmembrane conductance regulator (CFTR) protein. Among these, p.Phe508del is the most prevalent variant. The effects of this variant on the physiology of each tissue remains unknown. This study is aimed at predicting cell signaling pathways present in different tissues of fibrocystic patients, homozygous for p.Phe508del. The study involved analysis of two microarray datasets, E-GEOD-15568 and E-MTAB-360 corresponding to the rectal and bronchial epithelium, respectively, obtained from the ArrayExpress repository. Particularly, differentially expressed genes (DEGs) were predicted, protein-protein interaction (PPI) networks were designed, and centrality and functional interaction networks were analyzed. The study reported that p.Phe508del-mutated CFTR-allele in homozygous state influenced the whole gene expression in each tissue differently. Interestingly, gene ontology (GO) term enrichment analysis revealed that only "neutrophil activation" was shared between both tissues; however, nonshared DEGs were grouped into the same GO term. For further verification, functional interaction networks were generated, wherein no shared nodes were reported between these tissues. These results suggested that the p.Phe508del-mutated CFTR-allele in homozygous state promoted tissue-specific pathways in fibrocystic patients. The generated data might further assist in prediction diagnosis to define biomarkers or devising therapeutic strategies.
Asunto(s)
Fibrosis Quística/genética , Transducción de Señal/genética , Alelos , Biomarcadores/metabolismo , Epitelio/fisiología , Expresión Génica/genética , Homocigoto , Humanos , Mutación/genética , Mapas de Interacción de Proteínas/genética , Biología de Sistemas/métodosRESUMEN
Asexual reproduction in Trichoplax occurs mainly by binary fission and occasionally by the budding of epithelial spheres called "swarmers". The process that leads to binary fission and the mechanisms involved in this segregation are practically unknown. Trichoplax lacks a defined shape, presenting a constantly changing outline due to its continuous movements and body contractions. For this reason, and due to the absence of anatomical references, it has been classified as an asymmetric organism. Here, we report that a transient wound is formed in the marginal epithelium of the two new individuals produced by binary fission. By tracking the location of this epithelial wound, we can determine that successive dichotomous divisions are orthogonal to the previous division. We also found that LiCl paralyzes the cilia beating movement and body contractions and causes the placozoans to become circular in shape. This effect, as well as a stereotypic body folding behavior observed in detached placozoans and cell labeling experiments of the upper epithelium, indicate a cylindrical body symmetry for Placozoa.
Asunto(s)
Placozoa/fisiología , Animales , Cilios/fisiología , Epitelio/fisiologíaRESUMEN
Background and aims: Mice orally infected with T. gondii develop Crohn's disease (CD)-like enteritis associated with severe mucosal damage and a systemic inflammatory response, resulting in high morbidity and mortality. Previously, helminthic infections have shown therapeutic potential in experimental colitis. However, the role of S. mansoni in T. gondii-induced CD-like enteritis has not been elucidated. Our study investigated the mechanisms underlying T. gondii-induced ileitis and the potential therapeutic effect of S. mansoni coinfection. Methods: C57BL/6 mice were infected by subcutaneous injection of cercariae of the BH strain of S. mansoni, and 7-9 weeks later, they were orally infected with cysts of the ME49 strain of T. gondii. After euthanasia, the ileum was removed for histopathological analysis; staining for goblet cells; immunohistochemistry characterizing mononuclear cells, lysozyme expression, apoptotic cells, and intracellular pathway activation; and measuring gene expression levels by real-time PCR. Cytokine concentrations were measured in the serial serum samples and culture supernatants of the ileal explants, in addition to myeloperoxidase (MPO) activity. Results:T. gondii-monoinfected mice presented dense inflammatory cell infiltrates and ulcerations in the terminal ileum, with abundant cell extrusion, apoptotic bodies, and necrosis; these effects were absent in S. mansoni-infected or coinfected animals. Coinfection preserved goblet cells and Paneth cells, remarkably depleted in T. gondii-infected mice. Densities of CD4- and CD11b-positive cells were increased in T. gondii- compared to S. mansoni-infected mice and controls. MPO was significantly increased among T. gondii-mice, while attenuated in coinfected animals. In T. gondii-infected mice, the culture supernatants of the explants showed increased concentrations of TNF-alpha, IFN-gamma, and IL-17, and the ileal tissue revealed increased expression of the mRNA transcripts for IL-1 beta, NOS2, HMOX1, MMP3, and MMP9 and activation of NF-kappa B and p38 MAPK signaling, all of which were counterregulated by S. mansoni coinfection. Conclusion:S. mansoni coinfection attenuates T. gondii-induced ileitis by preserving mucosal integrity and downregulating the local inflammatory response based on the activation of NF-kappa B and MAPK. The protective function of prior S. mansoni infection suggests the involvement of innate immune mechanisms and supports a conceptually new approach to the treatment of chronic inflammatory diseases, including CD.
Asunto(s)
Coinfección/inmunología , Ileítis/prevención & control , Mucosa Intestinal/fisiopatología , Esquistosomiasis mansoni/inmunología , Terapia con Helmintos , Toxoplasmosis Animal/terapia , Animales , Apoptosis , Enfermedad de Crohn/terapia , Citocinas/sangre , Modelos Animales de Enfermedad , Regulación hacia Abajo , Epitelio/fisiología , Perfilación de la Expresión Génica , Ileítis/etiología , Ileítis/inmunología , Ileítis/patología , Sistema de Señalización de MAP Quinasas/fisiología , Ratones , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Peroxidasa/sangre , Síndrome de Respuesta Inflamatoria Sistémica/etiología , Síndrome de Respuesta Inflamatoria Sistémica/inmunología , Toxoplasmosis Animal/complicaciones , Toxoplasmosis Animal/inmunologíaRESUMEN
Androgens induce rat prostate induction from the urogenital sinus epithelium at embryonic day 17.5. Subsequent morphogenesis, including epithelial cord growth, branching, and canalization, results from concerted paracrine interactions with the stroma. A significant number of paracrine factors bind heparan sulfate (HS). We hypothesized that interfering with overall sulfation could disrupt the signaling mediated by HS-binding factors and that the undersulfated environment would allow investigation of individual exogenous morphogens. First, we investigated whether acinar morphogenesis involved HS-proteoglycan expression and found that syndecans 1 and 3 were upregulated in RWPE1 cells in the transition from two- to three-dimensional (3D) Matrigel, capable of promoting spheroid formation. We then investigated whether sodium chlorate, a general sulfation inhibitor, interfered with spheroid formation by RWPE1 cells and acinar morphogenesis in ex vivo ventral prostate (VP) organ culture. As expected, treatment with sodium chlorate inhibited spheroid formation by RWPE1 cells in 3D culture. Chlorate also inhibited ex vivo VP epithelial branching and canalization, resulting in long branchless epithelial structures. We then investigated whether the HS-binding factors, FGF10, TGFß1, and SDF1, could reverse the effect of sodium chlorate. Although no effect was seen in the FGF10- and TGFß1-treated samples, SDF1 promoted epithelial canalization in the low sulfated environment, highlighting its specific role in lumen formation. Altogether, the results show that sodium chlorate perturbed prostate morphogenesis and allowed investigation of factors involved in branching and/or canalization, implicating SDF1 signaling in epithelial canalization.
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Quimiocina CXCL12/metabolismo , Células Epiteliales/metabolismo , Morfogénesis/fisiología , Próstata/metabolismo , Próstata/fisiología , Animales , Línea Celular , Colágeno/metabolismo , Combinación de Medicamentos , Células Epiteliales/fisiología , Epitelio/metabolismo , Epitelio/fisiología , Factor 10 de Crecimiento de Fibroblastos/metabolismo , Regulación del Desarrollo de la Expresión Génica/fisiología , Proteoglicanos de Heparán Sulfato/metabolismo , Heparitina Sulfato/metabolismo , Humanos , Laminina/metabolismo , Masculino , Técnicas de Cultivo de Órganos/métodos , Organogénesis/fisiología , Proteoglicanos/metabolismo , Ratas , Ratas Wistar , Transducción de Señal/fisiología , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
The importance of juvenile hormone regulating insect oogenesis suggests looking for genes whose expression is regulated by this hormone. SPARC is a calcium-binding glycoprotein that forms part of the extracellular membranes, which in vertebrates participates in bones mineralization or regulating cell proliferation in some cancer types. This large number of functions described for SPARC in different species might be related to the significant differences in its structure observed when comparing different species-groups. Indeed, these structural differences allow characterizing the different clades. In the cockroach Blattella germanica, a SPARC homolog emerged from ovarian transcriptomes that were constructed to find genes responding to juvenile hormone. In insects, SPARC functions have been studied in oogenesis and in embryo development of Drosophila melanogaster. In the present work, using RNAi approaches, novel functions for SPARC in the B. germanica panoistic ovaries are described. We found that depletion of SPARC does not allow to the follicular cells to complete mitosis, resulting in giant follicular cells nuclei and in a great alteration of the ovarian follicle cytoskeleton. The SPARC contribution to B. germanica oogenesis occurs stabilizing the follicular cell program and helping to maintain the nuclear divisions. Moreover, SPARC is necessary to maintain the cytoskeleton of the follicular cells. Any modification of these key processes disables females for oviposition.
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Blattellidae/embriología , Citoesqueleto/metabolismo , Epitelio/fisiología , Oogénesis/fisiología , Osteonectina/metabolismo , Folículo Ovárico/embriología , Animales , Proliferación Celular , Femenino , Regulación del Desarrollo de la Expresión Génica , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Hormonas Juveniles/metabolismo , Mitosis/fisiología , Oogénesis/genética , Osteonectina/genética , Folículo Ovárico/citología , Folículo Ovárico/metabolismo , Interferencia de ARN , ARN Interferente Pequeño/genética , Vitelogeninas/biosíntesisRESUMEN
The current study aimed to determine if characteristics observed in vaginal cytology during the estrous cycle of female SYT cavies corresponded with proliferation of the vaginal epithelium, characterized by proliferating cell nuclear antigen (PCNA) immunolocalization, and with follicular development at different phases of the estrous cycle. After determining estrous cycle phases by vaginal cytology, females were euthanized at metestrus, diestrus, proestrus, and estrus. Histological study of the vaginal epithelium and ovary were then performed. Immunohistochemistry for PCNA in vaginal tissue at each cycle phase was also performed. Superficial cornified cells and early post-ovulatory follicles were found at estrus. Few nuclei below the enucleate superficial cells were immunoreactive to PCNA. At metestrus, the vaginal epithelium underwent desquamation and lost the superficial cornified cells; basal and intermediate cells appeared, and the post-ovulatory follicle formed an early corpus luteum. No PCNA immunoreactivity was observed. At diestrus, the corpus luteum was developed, and the vaginal epithelium contained basal and intermediate cells. There was PCNA immunoreactivity in the cellular nucleus in the germinative stratum of the epithelium. Because of the growth and maturation of ovarian follicles, the vaginal epithelium suffered intense proliferation at proestrus. Vaginal cytology revealed large intermediate cells and nucleated and enucleated superficial cornified cells. In the ovary, mature follicles were present. More apparent immunoreactivity of PCNA in the germinative layer was found. In summary, we inferred that vaginal exfoliative findings matched the proliferation process of the vaginal epithelium. PCNA immunolocalization occurred as well as corresponding follicular development in the ovaries.
Asunto(s)
Ciclo Estral/fisiología , Folículo Ovárico/citología , Folículo Ovárico/fisiología , Roedores/fisiología , Vagina/citología , Animales , Proliferación Celular , Diestro/fisiología , Epitelio/inmunología , Epitelio/fisiología , Estro/fisiología , Femenino , Técnicas Histológicas , Inmunohistoquímica , Folículo Ovárico/ultraestructura , Ovario/citología , Ovario/ultraestructura , Proestro/fisiología , Antígeno Nuclear de Célula en Proliferación/análisis , Antígeno Nuclear de Célula en Proliferación/inmunología , Roedores/anatomía & histología , Vagina/fisiologíaRESUMEN
Abstract:The tropical gar A. tropicus plays an important ecological role as it regulates other fish stocks in different water bodies in Southeastern México. Nevertheless, wild populations are declining, and one conservation alternative is the aquaculture production and basic knowledge of reproductive biology; for males, this requires the study of germ and somatic structures of testes, to characterize the reproductive cycle, and to provide basic knowledge for exploitation and conservation models and strategies. With this aim, a total of 24 males with an average sL = 47.2 cm were collected from wild populations from the Laguna Pomposú, municipality of Jalpa de Mendez (18°19' - 93°01'12" W), Tabasco, Mexico. Fish were collected with a trawl net and were transported live to the Tropical Aquaculture Laboratory, División Académica de Ciencias Biológicas (DACBiol), Universidad Juárez Autónoma de Tabasco (UJAT). Males were killed by prolonged immersion in MS222. Testes samples were collected from each specimen and were processed using the standard histological procedures, that consisted of dehydration in an ascending ethanol series, xylol, embedding in paraffin, sectioning at 7 µm, and staining with hematoxylin-eosin (HE). The diameter of 20 seminiferous tubules (Dst), height of germinal epithelium (Hge), gonadosomatic index (GSI) and gonad volume (gV) were determined monthly. Based on morphometric and morpho-physiological characteristics, the testes consisted of a network of anastomosed tubules with non-restricted cystic spermatogenesis, and a permanent germinal epithelium. This is the first report of a permanent germinal epithelium in A. tropicus. Five reproductive classes were histologically identified: Class I Regressed; Class II Early Maturation; Class III Mid Maturation; Class IV Late Maturation; Class V Regression. Monthly GSI, gV and Dst values were lower in January and February, the testis showed spermatozoa remains and a regenerating discontinuous germinal epithelium. In March spermiogenesis increased and proliferation of spermatogonia decreased. Male tropical gar followed a seasonal reproductive cycle, indicated by the monthly variation of the reproductive classes and the reproductive season processes observed, and for which temperature and rainfall seem to stimulate reproductive activity and spermiation. Rev. Biol. Trop. 64 (4): 1597-1609. Epub 2016 December 01.
Resumen:A. tropicus tiene un papel ecológico importante, como regulador de otras poblaciones de peces, en los cuerpos de agua de México, pero sus poblaciones silvestres se reducen. Una alternativa de conservación es el cultivo, el cual requiere caracterizar el ciclo reproductivo por medio del estudio de estructuras germinales y somáticas de los testículos, conocimientos que son básicos para formar modelos de aprovechamiento y conservación. Se capturaron mensualmente tres machos sexualmente maduros (N = 24), con un promedio de sL = 47.2 cm en Laguna de Pomposú, Jalpa de Méndez (18°19´59" N - 93°01´12" W), Tabasco, México, de octubre 2009 a septiembre 2010. La técnica de captura fue red de arrastre, se transportaron vivos al laboratorio de acuicultura tropical, DACBiol, UJAT. Los machos recolectados se sacrificaron con baños de inmersión en sobredosis de MS222, los testículos se procesaron para análisis histológico. Se determinó mensualmente el diámetro de 20 túbulos seminíferos (Dst), altura de epitelio germinal (Hge), índice gonadosomático (GSI) y volumen de gónada (gV). Características morfo-fisiológicas del testículo muestran que está constituido de una red de túbulos anastomosados con espermatogénesis quística no restringida, y un epitelio germinal permanente, de nuestro conocimiento es la primera vez que se reporta este tipo de epitelio en Holostei (Lepisosteiformes: Lepisosteidae). Se identificaron cinco clases reproductivas: Clase I Recrudescencia, Clase II Madurez temprana, Clase III Madurez intermedia, Clase IV Maduración tardía, Clase V Regresión, que al contrastarlo con el valor mensual de los indicadores sexuales "GSI, gV, Dst" muestra un patrón de variación; durante enero-febrero se presentan valores bajos, se observa un epitelio germinal discontinuo en regeneración; durante marzo se incrementa la proliferación de espermatogonias disminuyendo la espermatogénesis. Los machos de A. tropicus muestran una actividad reproductora estacional anual, explicado por las variaciones mensuales de los indicadores reproductores, donde la temperatura y la precipitación parecen tener un papel importante como factores que estimulan la actividad reproductora y por tanto la espermiación.
Asunto(s)
Animales , Masculino , Reproducción/fisiología , Testículo/anatomía & histología , Testículo/fisiología , Peces/fisiología , Valores de Referencia , Estaciones del Año , Maduración Sexual/fisiología , Espermatogénesis/fisiología , Espermatogonias/fisiología , Factores de Tiempo , Epitelio/fisiología , MéxicoRESUMEN
PURPOSE: Aging is associated with changes in the lung that leads to a decrease in its function. Alterations in structure and function in the small airways are well recognized in chronic lung diseases. The aim of this study was the assessment of cell turnover in the bronchiolar epithelium of mouse through the normal aging process. METHODS: Lungs from CD1 mice at the age of 2, 6, 12, 18, or 24 months were fixed in neutral-buffered formalin and paraffin-embedded. Proliferating cell nuclear antigen was examined by immunohistochemistry. Apoptosis was analyzed by in situ end-labeling of fragmented DNA. Epithelial dimensions were analyzed by morphometry. RESULTS: The 2-month-old mice showed significantly higher number of proliferating cells when compared with mice at all other age groups. The number of apoptotic cells in mice at 24 months of age was significantly greater than in mice at all other age groups. Thus, the number of epithelial cells decreased as the age of the subject increased. We also found reductions in both area and height of the bronchiolar epithelium in mice at 18 and 24 months of age. CONCLUSIONS: We found a decrease in the total number of epithelial cells in the aged mice, which was accompanied by a thinning of the epithelium. These changes reflect a dysregulated tissue regeneration process in the bronchiolar epithelium that might predispose to respiratory diseases in elderly subjects.
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Envejecimiento/fisiología , Bronquiolos/citología , Bronquiolos/fisiología , Células Epiteliales/fisiología , Epitelio/fisiología , Animales , Apoptosis , Proliferación Celular , Senescencia Celular , Epitelio/anatomía & histología , Masculino , Ratones , Antígeno Nuclear de Célula en Proliferación/metabolismoRESUMEN
The tropical gar A. tropicus plays an important ecological role as it regulates other fish stocks in different water bodies in Southeastern México. Nevertheless, wild populations are declining, and one conservation alternative is the aquaculture production and basic knowledge of reproductive biology; for males, this requires the study of germ and somatic structures of testes, to characterize the reproductive cycle, and to provide basic knowledge for exploitation and conservation models and strategies. With this aim, a total of 24 males with an average Lp = 47.2 cm were collected from wild populations from the Laguna Pomposú, municipality of Jalpa de Mendez (18°19' - 93°01'12" W), Tabasco, Mexico. Fish were collected with a trawl net and were transported live to the Tropical Aquaculture Laboratory, División Académica de Ciencias Biológicas (DACBiol), Universidad Juárez Autónoma de Tabasco (UJAT). Males were slaughtered by prolonged immersion in MS222. Testes samples were collected from each specimen and were processed using the standard histological procedures, that consisted of dehydration in an ascending ethanol series, xylol, embedding in paraffin, sectioning at 7 µm, and staining with hematoxylin-eosin (HE). The diameter of 20 seminiferous tubules (Tse), height of germinal epithelium (Egl), gonadosomatic index (IGS) and gonad volume (gV) were determined monthly. Based on morphometric and morpho-physiological characteristics, the testes consisted of a network of anastomosed tubules with non-restricted cystic spermatogenesis, and a permanent germinal epithelium. This is the first report of a permanent germinal epithelium in A. tropicus. Five reproductive classes were histologically identified: Class I Regressed; Class II Early Maturation; Class III Mid Maturation; Class IV Late Maturation; Class V Regression. Monthly GSI, gV and Tse values were lower in January and February, the testis showed spermatozoa remains and a regenerating discontinuous germinal epithelium. In March spermiogenesis increased and proliferation of spermatogonia decreased. Male tropical gar followed a seasonal reproductive cycle, indicated by the monthly variation of the reproductive classes and the reproductive season processes observed, and for which temperature and rainfall seem to stimulate reproductive activity and spermiation.
Asunto(s)
Peces/fisiología , Reproducción/fisiología , Testículo/anatomía & histología , Testículo/fisiología , Animales , Epitelio/fisiología , Masculino , México , Valores de Referencia , Estaciones del Año , Maduración Sexual/fisiología , Espermatogénesis/fisiología , Espermatogonias/fisiología , Factores de TiempoRESUMEN
The advanced glycation end products (AGEs) of proteins are common factors in the pathophysiology of a number of disorders related to aging. The skin generation of AGEs occurs mainly through nonenzymatic glycation reactions of extracellular matrix (ECM) proteins in the dermis. The AGEs have been touted as one of the factors responsible for healing impairment and loss of elasticity of healing skin, affecting growth, differentiation, and cellular motility, as well as cytokines response, metalloproteinases expression, and vascular hemostasis. In this study, we generated an in vitro full-thickness reconstructed skin based on a glycated collagen matrix dermal compartment to evaluate the effects of glycation on dermal ECM and ultimately on the epidermis. Epidermal differentiation and stratification patterns and the glycation-induced ECM changes were evaluated by histology, immunohistochemistry, and mRNA levels. In this study, we reported for the first time that changes in the dermal matrix caused by collagen I in vitro glycation processes also affect the epidermal compartment. We demonstrated that glycation of collagen induces expression of carboxymethyllysine in dermal and epidermal compartments and, consequently, an aging phenotype consisting of poor stratification of epidermal layers and vacuolization of keratinocyte cytoplasm. Increased expression of cell-cell adhesion markers, such as desmoglein and E-cadherin in glycated skins, is observed in the stratum spinosum, as well as an increased compression of dermal collagen matrix. We also submitted our 3D model of reconstructed glycated skin to screening of anti-AGE molecules, such as aminoguanidine, which prevented the glycated morphological status. Controlled human studies investigating the effects of anti-AGE strategies against skin aging are largely missing. In this context, we proposed the use of skin equivalents as an efficient model to investigate cellular interactions and ECM changes in the aging skin, and to elucidate the role of anti-AGEs molecules in this process.
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Envejecimiento de la Piel/patología , Piel/patología , Ingeniería de Tejidos , Animales , Diferenciación Celular , Epitelio/fisiología , Matriz Extracelular/metabolismo , Glicosilación , Humanos , Masculino , Microscopía , Ratas , Piel/anatomía & histologíaRESUMEN
PURPOSE: To evaluate the effects of angico bark extract (Anadenanthera colubrina var. cebil) in the healing process of the skin of rats.METHODS: Twenty adult male rats were divided into four groups of five animals each, according to the respective postoperative days, as follow: G4, G7, G14 and G21. Each group received two incisions on skin and subcutaneous tissue in the right and left antimere of the thoracic region, separated by a distance of 2 cm. The right lesion was treated daily with saline and the left with the angico alcoholic extract (5%). At the end of each experimental period, the animals were euthanized and fragments of the wound area with the edges were removed, fixed in 10% formaldehyde solution and processed for paraffin embedding. Histological sections (5 m of thickness) were stained with hematoxylin and eosin (HE), Gomori trichromic and picrosisirus red for morphological and morphometric analyses. Statistical analysis was done by ANOVA and Tukey-Kramer test (p 0.05).RESULTS: Morphological analysis showed larger fibroblasts and a higher concentration of collagen fibers in skyn wounds treated with the angico extract. Morphometric analysis demonstrated a significant increase in the number of fibroblasts at 7th and collagen in 7th and 14th days (p 0.01) in wounds treated with the angico extract.CONCLUSION: The angico alcoholic extract (Anadenanthera colubrina var. cebil) induces the acceleration of wound healing in skin wounds of rats.(AU)
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Animales , Masculino , Ratas , Epitelio/fisiología , Cicatrización de Heridas/efectos de los fármacos , Cicatrización de Heridas/fisiología , Fabaceae/química , Plantas Medicinales/química , Colágenos Fibrilares , Fibroblastos , FitoterapiaRESUMEN
Gingival wound healing comprises a series of sequential responses that allow the closure of breaches in the masticatory mucosa. This process is of critical importance to prevent the invasion of microbes or other agents into tissues, avoiding the establishment of a chronic infection. Wound healing may also play an important role during cell and tissue reaction to long-term injury, as it may occur during inflammatory responses and cancer. Recent experimental data have shown that gingival wound healing is severely affected by the aging process. These defects may alter distinct phases of the wound-healing process, including epithelial migration, granulation tissue formation, and tissue remodeling. The cellular and molecular defects that may explain these deficiencies include several biological responses such as an increased inflammatory response, altered integrin signaling, reduced growth factor activity, decreased cell proliferation, diminished angiogenesis, reduced collagen synthesis, augmented collagen remodeling, and deterioration of the proliferative and differentiation potential of stem cells. In this review, we explore the cellular and molecular basis of these defects and their possible clinical implications.
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Envejecimiento/fisiología , Encía/fisiología , Cicatrización de Heridas/fisiología , Tejido Conectivo/fisiología , Epitelio/fisiología , Humanos , Inflamación/fisiopatología , Péptidos y Proteínas de Señalización Intercelular/fisiología , Neovascularización Fisiológica/fisiologíaRESUMEN
Este trabajo muestra, desde el punto de vista de la normatividad de la Organización Panamericana de la Salud (OPS), el proceso de gestación, la metodología de implementación y los resultados obtenidos de la iniciativa de formación de recursos humanos en salud vía e-learning a través del Campus Virtual de Salud Pública de la Universidad de Guadalajara, México, a seis años de su inicio. Se trata de un informe especial del trabajo realizado por el comité institucional del campus virtual en la región occidental de México para generar un portal de Internet que se ajustara a los lineamientos del Modelo Estratégico establecido por el Nodo México y la OPS para la Región de las Américas. Este Campus Virtual inició sus actividades en el año 2007. Su filosofía es el uso de software libre y la colaboración entre instituciones. El nodo fue implementado en un año y ha logrado capacitar a más de 500 profesionales de la salud a través de cursos virtuales, su plataforma educativa y un repositorio de recursos virtuales de aprendizaje con interoperabilidad con los repositorios de México y de la Región de las Américas. El comité del Campus Virtual de la Universidad de Guadalajara ha intentado respetar lo más posible al modelo propuesto, lo que ha permitido cumplir la mayoría de los objetivos fijados en el plan de trabajo inicial, aunque ha enfrentado una serie de dificultades administrativas y de motivación de sus integrantes.
This paper discusses the gestation process, implementation methodology, and results obtained from the initiative to use e-learning to train human resources for health, six years after the launch of the Virtual Campus of Public Health of the University of Guadalajara (Mexico); the discussion is framed by Pan American Health Organization (PAHO) standards and practices. This is a special report on the work done by the institutional committee of the Virtual Campus in western Mexico to create an Internet portal that follows the guidelines of the strategic model established by Nodo México and PAHO for the Region of the Americas. This Virtual Campus began its activities in 2007, on the basis of the use of free software and institutional collaboration. Since the initial year of implementation of the node, over 500 health professionals have been trained using virtual courses, the node's educational platform, and a repository of virtual learning resources that are interoperable with other repositories in Mexico and the Region of the Americas. The University of Guadalajara Virtual Campus committee has followed the proposed model as much as possible, thereby achieving most of the goals set in the initial work plan, despite a number of administrative challenges and the difficulty of motivating committee members.
Asunto(s)
Animales , Perros , Hierro/toxicidad , Túbulos Renales/efectos de los fármacos , Adenilil Ciclasas/metabolismo , /metabolismo , División Celular/efectos de los fármacos , Línea Celular , Epitelio/efectos de los fármacos , Epitelio/patología , Epitelio/fisiología , Compuestos Férricos/toxicidad , Hierro/metabolismo , Túbulos Renales/patología , Túbulos Renales/fisiología , Células LLC-PK1 , Microscopía Electrónica , Porcinos , Cicatrización de Heridas/efectos de los fármacosRESUMEN
The exchange of substances between metazoan and the environment takes place across transporting epithelia that have two fundamental differentiated features: tight junctions (TJ) and apical/basolateral polarity. Usually, reviews of the structure and function of transporting epithelia follow a historical description of major biological findings, but seldom refer to the fact that it also required fundamental theoretical changes in the physics and chemistry involved. We make a brief description of the concatenation of both types of achievements, in which it becomes clear that the major source of conflicts was the enzyme Na(+),K(+)-ATPase (also referred to as "the pump"), because of its intrinsic mechanisms and its asymmetric expression on one side of epithelial cells only (polarity). This enzyme is also the receptor of the newly recognized hormone ouabain, whose chief function is to modulate cell contacts, such as TJs, several types of cell-cell contacts participating in polarization (as gauged through ciliogenesis).
Asunto(s)
Células Epiteliales/fisiología , Epitelio/fisiología , Ouabaína/metabolismo , Uniones Estrechas/fisiología , Transporte Biológico , Cilios/metabolismo , Claudina-2 , Humanos , Permeabilidad , ATPasa Intercambiadora de Sodio-PotasioRESUMEN
The vas deferens is a simple bioassay widely used to study the physiology of sympathetic neurotransmission and the pharmacodynamics of adrenergic drugs. The role of ATP as a sympathetic co-transmitter has gained increasing attention and furthered our understanding of its role in sympathetic reflexes. In addition, new information has emerged on the mechanisms underlying the storage and release of ATP. Both noradrenaline and ATP concur to elicit the tissue smooth muscle contractions following sympathetic reflexes or electrical field stimulation of the sympathetic nerve terminals. ATP and adenosine (its metabolic byproduct) are powerful presynaptic regulators of co-transmitter actions. In addition, neuropeptide Y, the third member of the sympathetic triad, is an endogenous modulator. The peptide plus ATP and/or adenosine play a significant role as sympathetic modulators of transmitter's release. This review focuses on the physiological principles that govern sympathetic co-transmitter activity, with special interest in defining the motor role of ATP. In addition, we intended to review the recent structural biology findings related to the topology of the P2X1R based on the crystallized P2X4 receptor from Danio rerio, or the crystallized adenosine A2A receptor as a member of the G protein coupled family of receptors as prototype neuro modulators. This review also covers structural elements of ectonucleotidases, since some members are found in the vas deferens neuro-effector junction. The allosteric principles that apply to purinoceptors are also reviewed highlighting concepts derived from receptor theory at the light of the current available structural elements. Finally, we discuss clinical applications of these concepts.
Asunto(s)
Unión Neuroefectora/fisiología , Conducto Deferente/fisiología , Animales , Epitelio/fisiología , Humanos , Masculino , Unión Neuroefectora/anatomía & histología , Receptores Purinérgicos/metabolismo , Conducto Deferente/anatomía & histologíaRESUMEN
The reproductive system of some fish species presents elaborate mechanisms by which the females store spermatozoa inside their ovaries, keeping them viable for fertilization for an extended period of time. However, as intriguing as this sperm storage is, it is not yet understood how the sperm can remain viable in the ovary. Aiming to understand this phenomenon, the epithelium covering the ovarian lamellae, that is, the germinal epithelium, of the Cangati (Trachelyopterus galeatus), an inseminating catfish, was evaluated taking into account the different stages of the annual reproductive cycle. The germinal epithelium morphology changed during the annual reproductive cycle, presumably in preparation to receive the spermatozoa and keep them viable until fertilization. There was a progressive increase of the epithelium height. Also the number of intercellular junctions, desmosomes, and extended tight junctions, apparently increased forming chains that could be regarded as a barrier to isolate the sperm from the female immune system. Synthetic organelles were active releasing cytoplasmic granules and secretion in the epithelial enfolds in which the spermatozoa were deeply embedded. Concomitantly, oogonium nests were formed in the germinal epithelium during early folliculogenesis.
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Bagres/fisiología , Ovario/fisiología , Espermatozoides/citología , Animales , Bagres/anatomía & histología , Supervivencia Celular , Epitelio/fisiología , Femenino , Fertilización , Inseminación , Masculino , Ovario/anatomía & histología , Espermatozoides/fisiologíaRESUMEN
PURPOSE: To assess the viability of cultured epithelium and preserved by freezing for periods varying from one month to one year. METHODS: Samples of cultured epithelium were incubated in cryoprotectant medium (Group A), packed in aluminum envelopes and packed in polystyrene boxes. The boxes were subjected to a temperature of-70 ºC. After freezing for a period of time ranging from one to 12 months, cultured epithelial samples were assessed for their viability by vital staining (Trypan blue) and metabolic analysis based on glucose consumption and lactate production. Samples of not frozen cultured epithelium (Group B) were also tested for viability and the results obtained were used as comparison parameter for the variation of viability. RESULTS: Statistical analysis between the group A and B indicate that the mean age of the donors (p=0.51) and the culture time (p=1.18) showed no statistical difference. In 30 days we obtained 37% of the original viability of cultured epithelium, 25% at six months and one year, less than 15%. This trend was confirmed statistically with a reduction of approximately 1.8% of the original viability epithelium cultured every 30 days of storage. In the analysis by lactate production, similar results were observed. In the analysis by the glucose consumption results were not significant. The viability indices show statistically significant difference between the group A and B (p<0.0001). CONCLUSIONS: Although cryopreserved cultured epithelium showed significant reduction of viability, all samples remained viable. It was also found that the viability of cryopreserved cultured epithelial decreased as a function of storage time.
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Criopreservación/métodos , Piel , Supervivencia Tisular/fisiología , Adulto , Aloinjertos/fisiología , Supervivencia Celular/fisiología , Células Cultivadas/fisiología , Crioprotectores/farmacología , Medios de Cultivo , Epitelio/fisiología , Femenino , Humanos , Masculino , Valores de Referencia , Reproducibilidad de los Resultados , Estadísticas no Paramétricas , Factores de Tiempo , Adulto JovenRESUMEN
PURPOSE: To assess the viability of cultured epithelium and preserved by freezing for periods varying from one month to one year. METHODS: Samples of cultured epithelium were incubated in cryoprotectant medium (Group A), packed in aluminum envelopes and packed in polystyrene boxes. The boxes were subjected to a temperature of-70ºC. After freezing for a period of time ranging from one to 12 months, cultured epithelial samples were assessed for their viability by vital staining (Trypan blue) and metabolic analysis based on glucose consumption and lactate production. Samples of not frozen cultured epithelium (Group B) were also tested for viability and the results obtained were used as comparison parameter for the variation of viability. RESULTS: Statistical analysis between the group A and B indicate that the mean age of the donors (p=0.51) and the culture time (p=1.18) showed no statistical difference. In 30 days we obtained 37% of the original viability of cultured epithelium, 25% at six months and one year, less than 15%. This trend was confirmed statistically with a reduction of approximately 1.8% of the original viability epithelium cultured every 30 days of storage. In the analysis by lactate production, similar results were observed. In the analysis by the glucose consumption results were not significant. The viability indices show statistically significant difference between the group A and B (p<0.0001). CONCLUSIONS: Although cryopreserved cultured epithelium showed significant reduction of viability, all samples remained viable. It was also found that the viability of cryopreserved cultured epithelial decreased as a function of storage time.
Asunto(s)
Adulto , Femenino , Humanos , Masculino , Adulto Joven , Criopreservación/métodos , Piel , Supervivencia Tisular/fisiología , Aloinjertos/fisiología , Medios de Cultivo , Supervivencia Celular/fisiología , Células Cultivadas/fisiología , Crioprotectores/farmacología , Epitelio/fisiología , Valores de Referencia , Reproducibilidad de los Resultados , Estadísticas no Paramétricas , Factores de TiempoRESUMEN
The feral pig (Sus scrofa sp) also known as Monteiro pig, originated from a domestic pig breed that was introduced into Pantanal region in Brazil in the eighteenth century. Although the feral pig has commercial potential, there are few reports in the literature concerning the reproductive biology of this species. Therefore, the aim of this study was to further describe the feral pig testis parenchyma as well as characterize the stages of the seminiferous epithelium cycle by tubular morphology method, and to evaluate the number of differentiated spermatogonia generations in this species. Eight sexually mature feral pigs were analyzed. Fragments of testes were embedded in plastic resin and used to prepare slides for morphometrical studies. It was concluded that the feral pig has six generations of differentiated spermatogonials (A1, A2, A3, A4, In, B) and that the cellular composition in the eight stages of the seminiferous epithelium cycle of these animals were very similar to those reported in species of suidae and tayssuidae already studied.