Increase In Intercellular Space Research Articles

Effects of apelin on neonatal brain neurogenesis in L-NAME-induced maternal preeclampsia

The aim of this study was to investigate the possible protective effects of apelin, which is known to have antioxidant and anti-inflammatory effects, on changes in neurogenesis in newborns of pregnant rats with L-NAME-induced preeclampsia. Wistar albino female rats were divided into four experimental groups: Control, Apelin, Preeclampsia and Preeclampsia + Apelin. Blood pressure was measured on the 5th, 11th and 17th days of gestation, urine protein was analyzed from urine samples collected for 24 h on the 6th, 12th and 18th days and serum creatinine was analyzed from serum samples. Maternal kidney and placenta tissues were obtained to establish the preeclampsia model, and neonatal brain tissues including the cortex, hippocampus and cerebellum regions were obtained to investigate neurogenesis and examined by histological and immunohistochemical methods. The number of newborns, body weight and brain weight of the newborns were measured. eNOS, IL-10, nNOS and NO levels in the brain analyzed via ELISA. Mean arterial pressure, urine protein and serum creatinine increased in the preeclampsia. Newborn weight decreased in the Preeclampsia group, the values in the Preeclampsia + Apelin group were closer to the Control and Apelin groups. In the Preeclampsia group, edema and dilatation in the proximal and distal tubules of kidneys, perivillous fibrin deposition and increase in syncytial nodules of placenta were observed. VEGF immunoreactivity decreased and iNOS immunoreactivity increased in both kidney and placenta. In neonatal brain tissue examinations, cytotoxic edema accompanied by thinning of cortex, delayed migration and lower cell counts in the hippocampus, and increase in intercellular spaces and EGL thickening in the cerebellum were observed in the preeclampsia. Expression of NeuN, GFAP, MBP, IL-10, eNOS, nNOS and NO levels decreased, whereas expression of Iba-1 increased in the preeclampsia. In the Preeclampsia + Apelin group, these findings were similar to the Control and Apelin groups. Apelin administration was found to be beneficial for preventing the adverse consequences of preeclampsia, but further experimental and clinical studies are needed to better understand these effects.

Aspergillus Fumigatus Spore Proteases Alter the Respiratory Mucosa Architecture and Facilitate Equine Herpesvirus 1 Infection.

Numerous Aspergillus fumigatus (Af) airborne spores are inhaled daily by humans and animals due to their ubiquitous presence. The interaction between the spores and the respiratory epithelium, as well as its impact on the epithelial barrier function, remains largely unknown. The epithelial barrier protects the respiratory epithelium against viral infections. However, it can be compromised by environmental contaminants such as pollen, thereby increasing susceptibility to respiratory viral infections, including alphaherpesvirus equine herpesvirus type 1 (EHV-1). To determine whether Af spores disrupt the epithelial integrity and enhance susceptibility to viral infections, equine respiratory mucosal ex vivo explants were pretreated with Af spore diffusate, followed by EHV-1 inoculation. Spore proteases were characterized by zymography and identified using mass spectrometry-based proteomics. Proteases of the serine protease, metalloprotease, and aspartic protease groups were identified. Morphological analysis of hematoxylin-eosin (HE)-stained sections of the explants revealed that Af spores induced the desquamation of epithelial cells and a significant increase in intercellular space at high and low concentrations, respectively. The increase in intercellular space in the epithelium caused by Af spore proteases correlated with an increase in EHV-1 infection. Together, our findings demonstrate that Af spore proteases disrupt epithelial integrity, potentially leading to increased viral infection of the respiratory epithelium.

Dynamic Prediction Model for Initial Apple Damage.

Prediction models of damage severity are crucial for the damage expression of fruit. In light of issues such as the mismatch of existing models in actual damage scenarios and the failure of static models to meet research needs, this article proposes a dynamic prediction model for damage severity throughout the entire process of apple damage and studies the relationship between the initial bruise form and impact energy distribution of apple damage. From the experiments, it was found that after impact a "cell death zone" appeared in the internal pulp of the damaged part of Red Delicious apples. The reason for the appearance of the cell death zone was that the impact force propagated in the direction of the fruit kernel in the form of stress waves; the continuous action of which continuously compressed the pulp's cell tissue. When the energy absorbed via elastic deformation reached the limit value, intercellular disadhesion of parenchyma cells at the location of the stress wave peak occurred to form cell rupture. The increase in intercellular space for the parenchyma cells near the rupture site caused a large amount of necrocytosis and, ultimately, formed the cell death zone. The depth of the cell death zone was closely related to the impact energy. The correlation coefficient r between the depth of the cell death zone and the distribution of impact energy was slightly lower at the impact height of 50 mm. As the impact height increased, the correlation coefficient r increased, approaching of value of 1. When the impact height was lower (50 mm), the correlation coefficient r had a large distribution range (from 0.421 to 0.983). As the impact height increased, the distribution range significantly decreased. The width of the cell death zone had a poor correlation with the pressure distribution on the impact surface of the apples that was not related to the impact height. In this article, the corresponding relationship between the form and impact energy distribution of the internal damaged tissues in the initial damage of Red Delicious apples was analyzed. This analysis aimed to provide a research concept and theoretical basis for more reliable research on the morphological changes in the damaged tissues of apples in the future, further improving the prediction accuracy of damage severity.

Effect of ligustrazine on hypoxic-ischemic encephalopathy in neonatal rats by regulating autophagy through the PINK1/Parkin pathway

To study the effect of ligustrazine injection on mitophagy in neonatal rats with hypoxic-ischemic encephalopathy (HIE) and its molecular mechanism. Neonatal Sprague-Dawley rats, aged 7 days, were randomly divided into a sham-operation group with 8 rats, a model group with 12 rats, and a ligustrazine group with 12 rats. The rats in the model group and the ligustrazine group were used to establish a neonatal rat model of HIE by ligation of the left common carotid artery followed by hypoxia treatment, and blood vessels were exposed without any other treatment for the rats in the sham-operation group. The rats in the ligustrazine group were intraperitoneally injected with ligustrazine (20 mg/kg) daily after hypoxia-ischemia, and those in the sham-operation group and the model group were intraperitoneally injected with an equal volume of normal saline daily. Samples were collected after 7 days of treatment. Hematoxylin and eosin staining and Nissl staining were used to observe the pathological changes of neurons in brain tissue; immunohistochemical staining was used to observe the positive expression of PINK1 and Parkin in the hippocampus and cortex; TUNEL staining was used to measure neuronal apoptosis; Western blotting was used to measure the expression levels of the mitophagy pathway proteins PINK1 and Parkin and the autophagy-related proteins Beclin-1, microtubule-associated protein 1 light chain 3 (LC3), and ubiquitin-binding protein (P62). Compared with the sham-operation group, the model group had a significant reduction in the number of neurons, an increase in intercellular space, loose arrangement, lipid vacuolization, and a reduction in Nissl bodies. The increased positive expression of PINK1 and Parkin, apoptosis rate of neurons, and protein expression levels of PINK1, Parkin, Beclin1 and LC3 (P<0.05) and the decreased protein expression level of P62 in the hippocampus were also observed in the model group (P<0.05). Compared with the model group, the ligustrazine group had a significant increase in the number of neurons with ordered arrangement and an increase in Nissl bodies, significant reductions in the positive expression of PINK1 and Parkin, the apoptosis rate of neurons, and the protein expression levels of PINK1, Parkin, Beclin1, and LC3 (P<0.05), and a significant increase in the protein expression level of P62 (P<0.05). Ligustrazine can alleviate hypoxic-ischemic brain damage and inhibit neuronal apoptosis in neonatal rats to a certain extent, possibly by inhibiting PINK1/Parkin-mediated autophagy.

Clinical and morphophysiological hepatic alterations caused by the ingestion of food with added ricinoleic acid esters a bioactive with acaricidal potential

The present study investigated the effects of castor oil ricinoleic acid through analysis of blood biochemical parameters and the liver morphology of female rabbits (animal model). The results of Bioassay 1 and Bioassay 2 showed that the reference indexes of the hepatic enzymes alkaline phosphatase, gamma-glutamyl transferase, and alanine aminotransferase were not significantly altered. However, the hepatic enzyme aspartate aminotransferase of the rabbits tested in Bioassay 2 presented alterations, suggesting a hepatotoxic effect of the esters. The morphological and histological data showed that the liver of the rabbits fed with the ester-added food presented the following alterations: a) disorganization of the hepatic cords; b) an increase in intercellular spaces; c) hepatocytes with barely evident cell limits; and d) hepatocytes with irregular and pyknotic nuclei, in most cases displaced to the periphery of the cell due to extensive vacuolation. The histochemical analysis showed that the hepatocytes of the rabbits fed with commercial food with added esters had a decrease in lipid content and an increase in the levels of stored cytoplasmic glycogen.

Effects of postharvest ripening on physicochemical properties, microstructure, cell wall polysaccharides contents (pectin, hemicellulose, cellulose) and nanostructure of kiwifruit (Actinidia deliciosa)

Kiwifruit undergoes rapid softening and quality changes after harvest, which has an important impact on consumer appeal, shelf life and market value. Effects of postharvest ripeness on physicochemical properties, microstructure, cell wall fraction (pectin, hemicellulose and cellulose) contents and nanostructure, and pectin molecular weight were investigated. As the postharvest time increased from 0 to 6 days, the contents of total soluble solid and water-soluble pectin increased by 77.4% and 113.0%, respectively; while the content of starch, water insoluble pectin, and hemicellulose decreased by 89.9%, 46.5%, and 33.5%, respectively. Disappearance of starch granules and the increase of intercellular space were observed using SEM. AFM analysis showed that both hemicellulose and pectin depolymerized gradually during storage. Measurements of the molecular weight confirmed the occurrence of depolymerization and degradation of pectin, while the cellulose content, nanostructure, and diameter of individual microfibrils did not change significantly. The findings in current work revealed the quality evolution and related mechanism of kiwifruit during storage.

Effect of electroacupuncture on tissue elasticity and expression of muscular MyoD and myogenin in rats with acute gastrocnemius contusion

To observe the effect of electroacupuncture (EA) at "Ashi" acupoint and "Kunlun" (BL60) on elastic modulus, histopathological changes and expression of myogenic regulatory factors in gastrocnemius(GM) contusion rats, so as to explore the therapeutic effect of local acupoint selection and acupoint selection along channel. Male SD rats were randomly divided into blank control (n＝5), model (n＝15), Ashi-point (n＝15) and BL60 (n＝15) groups. The acute GM contusion model was established by striking (free falling) the GM with a homemade hitter. EA (0.5 to 1.0 mA, 2 Hz/10 Hz) was applied to Ashi-point (local focus) and BL60 for 30 min 24 h after muscle injury. The elasticity maximum (Emax) of gastrocnemius muscle was measured by using an ultrasonic device. Histopathological changes were observed after H．E. stain, and the number of Myogenic differentiation(MyoD)- and Myogenin (MyoG)-positive cells was detected by using immunohistochemistry. After mdeling, the Emax value of GM was significantly increased from the 3rd h to 7th day in comparison with pre- injury of muscle (P<0.05), and was markedly increased on the 3rd day and obviously lower on day 7 in the Ashi-point group than in the model group (P<0.05). The numbers of MyoD- and MyoG-positive cells of GM were significantly increased on day 7 in the model group than in the blank control group (P<0.05), and both further increased in Ashi-point on day 3 and 5, and MyoG-positive cells further increased in BL60 group on day 5 and in Ashi-point group on day 7 relevant to the model group(P<0.05). The therapeutic effect of EA-Ashi-point was apparently superior to that of BL60 in up-regulating Emax on day 3 and in up-regulating the number of MyoD-positive cells on day 3 and 5 (P<0.05). H．E. stain showed disordered arrangement of muscle fibers, infiltration of inflammatory cells, increase of intercellular space, and edema on day 3 after modeling (which was milder in the Ashi-point group); and gradual fusion and thickening of new born muscle fibers with obvious connective tissue hyperplasia converged to the lesioned region on day 7 in the model group (convergence of new born muscle cells to the lesion region in both EA groups, and more complete tissues in the Ashi-point group). EA of Ashi-point and BL60 can up-regulate the expression of myogenic regulatory factors MyoD and MyoG of GM tissue in GM contusion rats, which may contribute to its function in promoting recovery of muscle elasticity. The role of EA-Ashi-point is superior to that of EA-BL60.

Effect of Smartphone Light Fluxes on Cornea: A Biophysical Study.

Objective:Biophysical study to investigate (a) the effects of smartphone light fluxes (SPLF) on isolated mammalian cornea and model protein (insulin), (b) to predict the possible visual interference of SPLF.Materials and Methods:Fresh goat cornea and insulin protein were used as an experimental model system. The energy of absorbed SPLF was measured using chemical dosimeter. The effect of SPLF on the aggregation of model protein was studied using fluorescence spectroscopy and dynamic light scattering (DLS). Fluorescence microscopy, scanning electron microscopy (SEM), DLS, were used for cornea imaging.Results:The spectral emission peak of SPLF was observed at 380 nm and 420 nm. Absorbed radiation of SPLF was found to be 2.82 mWm-2 and 1.92 mWm-2 for collimated (focussed) and noncollimated (nonfocussed) condition, respectively. Secondary structural changes of insulin were observed by fluorescence and zeta potential after SPLF exposure. SEM study revealed the disorganization of the epithelial cell surface, increase in intercellular space, disorganization of primary epithelium layer, and exposure of the second layer is seen in depth. Differential Interference Microscopy showed an optical gradient in images that appears to be changed in specimen structure. Fluorescence microscopy showed disorganization in epithelial cell pattern. A significant difference in bio-molecular permeation was observed in the exposed cornea. Ultraviolet UV-visible spectroscopy study indicated a reduction in light transmission through the cornea.Conclusions:The obtained results indicate changes in physicochemical and morphological modifications in the cornea and insulin modifications after exposed to SPLF.

Morpho-anatomical, physiological and biochemical adaptive responses to saline water of Bougainvillea spectabilis Willd. trained to different canopy shapes

Bougainvillea is widely used as potential ornamental shrub in revegetation and landscaping projects as well as in urban landscape design in Mediterranean areas characterized by saline and sodic soils. The current research aimed at elucidating the morphological, physiological and biochemical changes occurring in Bougainvillea spectabilis ‘Alexandra’ grown under salt stress conditions (1 mM [S1, electrical conductivity EC = 2.0 dS m−1] non-salt control, 40 mM [S40, EC = 6.5 dS m−1] and 80 mM [S80, EC = 11.0 dS m−1] NaCl). The response of plants trained to two canopy shapes (globe [GSh] and pyramid [PSh]) has been compared in order to unravel possible adaptive mechanisms, combining morpho-functional traits at different levels, to cope with saline water. PSh plants under moderate saline water (40 mM NaCl) showed higher leaf dry weight, leaf number and area as well as higher number of flowers per plant than those observed in GSh Bougainvillea plants. Saline water-triggered leaf anatomical changes were also harmonized to physiological adjustments in both canopy shapes in such a way to counteract the reduction in transpiration due to stomatal limitations by adopting traits improving CO2 diffusion in the mesophyll and investing more resources in the assimilatory tissue (e.g. increase in intercellular spaces and incidence of palisade parenchyma over the total mesophyll thickness). Bougainvillea plants enact many-sided strategies to acclimate to S80 salinity by harmonizing several mechanisms including sodium and chloride inclusion in leaves and their compartmentalization in vacuoles, retention of high levels of potassium in the cytosol to osmotically balance the cytoplasm and other organelles within the vacuole, modulation of leaf functional anatomical traits, in addition to a C2 Kranz mechanism of photosynthesis which allow plants to successfully cope with stress.

Investigation of conjunctivochalasis histopathology with light and electron microscopy in patients with conjunctivochalasis in different locations.

To investigate changes in conjunctival tissue of conjunctivochalasis (CCh) patients and to determine the relationship between pathological findings and localization of loose conjunctiva. Our study included nineteen eyes of 19 patients who were referred to Cukurova University Ophthalmology Department based on ocular surface symptoms and CCh detected in ocular examination. Amniotic membrane was applied after conjunctival excision as surgical treatment. The control group was formed with five eyes of five patients who are similar in terms of age and gender distribution with our study group. Tissue samples obtained from the study and control groups were investigated with light and electron microscopy. Results of pathological examination of conjunctival tissues revealed increased inflammation in 13 patients (68%), lymphatic ectasia in 12 patients (63%), and loss of goblet cells in 17 patients (89%). Destruction of elastic fibers was detected in all cases by staining with elastic van Gieson. After semiquantitative assessment, varying degrees of light microscopic findings were noted considering the localization of CCh. No statistically significant relationship was observed between light microscopic findings and CCh location (p > 0.05 for all). Electron microscopic investigation revealed increase in intercellular spaces, increased cytoplasmic electron density, and the presence of slight vacuolization in cell cytoplasm, and heterochromatin clumping in nuclei of cells in conjunctival samples. Mechanical and inflammatory factors induce development of CCh, and signs associated with these factors can be detected with light and electron microscopy of conjunctival tissue. No relationship was observed between CCh localization and pathological changes in tissues examined in our study, and large-scale case series are required to evaluate the possible effect of CCh localization on pathological findings.

Desuckering effect of KH2PO4 mixed with paclobutrazol and its influence on banana (Musa paradisiaca AA) mother plant growth

Abstract This study investigated the desuckering effect of a 2:1 mixture of KH2PO4 (KDP) and paclobutrazol (PBZ) and its influence on banana mother plant growth, fruit yield, and quality. The mixed reagent was injected into suckers sprouting from 5-month-old banana (Musa paradisiaca AA) plants after removing the above-ground part of the suckers. Compared to control plants, treatment with the KDP/PBZ mixture significantly reduced superoxide dismutase, peroxidase, and catalase activities but increased malondialdehyde content at sucker growth points and in adjacent tissue after 1 d. There was evidence of damage to the cellular structure at sucker growth points after 2 d of treatment, with an increase in intercellular space as well as rupturing and death of most cells. All suckers died 3 d later, resulting in a mortality rate of up to 100%, and there was no subsequent regrowth of the dead suckers; indicating that a single application of the KDP/PBZ reagent is a viable desuckering practice. Relative to the control, the treatment of suckers also indirectly increased the height and stem diameter of mother plants, which was associated with increased fruit yield (yield per plant, weight per hand, weight per fruit, transverse and longitudinal diameters of fruit fingers) and enhanced nutritional quality of the fruits (edible fraction, soluble solid, titratable acidity, vitamin C, and moisture content). These results indicate that a 2:1 KDP/PBZ mixture can effectively remove banana suckers while improving fruit yield and the quality of the mother plant. The proposed desuckering practice can help to address challenges associated with banana production such as difficulties in desuckering and preventing sucker regrowth, minimizing fertilizer waste, and high costs.

Global Gene Expression Profile of the Hippocampus in a Rat Model of Vascular Dementia.

Vascular dementia (VD) has been one of the most serious public health problems worldwide. It is well known that cerebral hypoperfusion is the key pathophysiological basis of VD, but it remains unclear how global genes in hippocampus respond to cerebral ischemia-reperfusion. In this study, we aimed to reveal the global gene expression profile in the hippocampus of VD using a rat model. VD was induced by repeated occlusion of common carotid arteries followed by reperfusion. The rats with VD were characterized by deficit of memory and cognitive function and by the histopathological changes in the hippocampus, such as a reduction in the number and the size of neurons accompanied by an increase in intercellular space. Microarray analysis of global genes displayed up-regulation of 7 probesets with genes with fold change more than 1.5 (P < 0.05) and down-regulation of 13 probesets with genes with fold change less than 0.667 (P < 0.05) in the hippocampus. Gene Ontology (GO) and pathway analysis showed that the up-regulated genes are mainly involved in oxygen binding and transport, autoimmune response and inflammation, and that the down-regulated genes are related to glucose metabolism, autoimmune response and inflammation, and other biological process, related to memory and cognitive function. Thus, the abnormally expressed genes are closely related to oxygen transport, glucose metabolism, and autoimmune response. The current findings display global gene expression profile of the hippocampus in a rat model of VD, providing new insights into the molecular pathogenesis of VD.

Liver histology and ultrastructure of the Italian newt (Lissotriton italicus): Normal structure and modifications after acute exposure to nonylphenol ethoxylates

We examined, from a morphological and ultrastructural point of view, the liver of the Italian newt (Lissotriton italicus), under basal conditions and after exposure to nonylphenol ethoxylates (NPEs). Nonylphenol ethoxylates are surfactants widely used in a variety of industrial and agricultural processes that may pose a significant risk to aquatic fauna. NPEs, and their degradation intermediates, are known to affect reproductive biology acting as endocrine disruptors; besides estrogenic effects, nonylphenolic compounds may induce organ toxicity, particularly in liver and gonads. We investigated the effects of a nonylphenol ethoxylate (NPE10-ETO) on L. italicus liver using two low concentrations, consistent with the environmental concentrations. For this purpose, animals were exposed to nominal concentrations of 50 and 100μg/L in a short-term experiment (96h). A morpho-functional analysis was performed in order to investigate the amphibian responses to NPEs thus contributing to elucidate other potential mode of action of these compounds; indeed very little attention has been dedicated to amphibians though they are often exposed to such contaminants in aquatic ecosystems. Pathological alterations on liver histology and ultrastructure were observed at both tested concentrations; the main effects recorded were: increase of intercellular spaces, accumulation of large lipid droplets, increase in melanin content, and a degeneration phenomenon. We also detected, through confocal analysis, the induction of caspase-3, a key mediator of apoptosis, and an up-regulation of cytochrome P450-1A. By using both ultrastructural and a morpho-functional approach, we found that a short-term exposure to NPEs negatively affected the amphibian liver.

Scanning Electron Microscopic Study of Microstructure of Gala Apples During Hot Air Drying

Microscopic changes that occur in plant food materials during drying significantly influence the macroscopic properties and quality factors of the dried food materials. It is critical to study the microstructure to understand the underlying cellular mechanisms to improve the performance of food drying techniques. However, there is limited research on such microstructural changes of plant food material during drying. In this work, Gala apple parenchyma tissue samples were studied using a scanning electron microscope for gradual microstructural changes as affected by temperature, time, and moisture content during hot air drying at two drying temperatures: 57 and 70°C. For fresh samples, the average cellular parameter values were as follows: cell area, 20,000 µm2; ferret diameter, 160 µm; perimeter, 600 µm; roundness, 0.76; elongation, 1.45; and compactness, 0.84. During drying, a higher degree of cell shrinkage was observed with cell wall warping and an increase in intercellular space. However, no significant cell wall breakage was observed. The overall reductions in cell area, ferret diameter, and perimeter were about 60, 40, and 30%. The cell roundness and elongation showed overall increments of about 5% and the compactness remained unchanged. Throughout the drying cycle, cellular deformations were mainly influenced by the moisture content. During the initial and intermediate stages of drying, cellular deformations were also positively influenced by the drying temperature and the effect was reversed at the final stages of drying, which provides clues regarding case hardening of the material.

Exposure and post-exposure effects of endosulfan on Bufo bufo tadpoles: Morpho-histological and ultrastructural study on epidermis and iNOS localization

Endosulfan is a persistent organic pollutant (POP) that has lethal and sublethal effects on non-target organisms, including amphibians. In a laboratory study, we investigated direct and post-exposure effects of endosulfan on Bufo bufo tadpoles. For this purpose we exposed the tadpoles to a single short-term contamination event (96h) at an environmentally-realistic concentration (200μg endosulfan/L). This was followed by a recovery period of 10 days when the experimental animals were kept in pesticide-free water. The endpoints were assessed in terms of mortality, incidence of deformity, effects on behavior, and the morpho-functional features of the epidermis. We found that a short-term exposure to the tested concentration of endosulfan did not cause mortality but induced severe sublethal effects, such as hyperactivity, convulsions, and axis malformations. Following relocation to a pesticide-free environment, we noted two types of response within the experimental sample, in terms of morphological and behavioral traits. Moreover, by using both ultrastructural and a morpho-functional approach, we found that a short-term exposure to endosulfan negatively affected the amphibian epidermis. We also observed several histo-pathological alterations: increased mucous secretion, an increase in intercellular spaces and extensive cell degeneration, together with the induction of an inducible isoform of nitric oxide synthase (iNOS).Following the post-exposure period, we found large areas of epidermis in which degeneration phenomena were moderate or absent, as well as a further increase in iNOS immunoreactivity. Thus, after 10 days in a free-pesticide environment, the larval epidermis was able to partially replace elements that had been compromised due to a physiological and/or a pathological response to the pesticide. These results highlight the need for both exposure and post-exposure experiments, when attempting to assess pollutant effects.

257 TRPV4 IS INVOLVED IN CELL JUNCTION FORMATION IN THE UROGENITAL TRACT. AN ULTRASTRUCTURAL STUDY

You have accessJournal of UrologyBladder and Urethra: Anatomy, Physiology and Pharmacology I1 Apr 2012257 TRPV4 IS INVOLVED IN CELL JUNCTION FORMATION IN THE UROGENITAL TRACT. AN ULTRASTRUCTURAL STUDY Dick Janssen, Kees Jansen, John Heesakkers, and Jack Schalken Dick JanssenDick Janssen Nijmegen, Netherlands More articles by this author , Kees JansenKees Jansen Nijmegen, Netherlands More articles by this author , John HeesakkersJohn Heesakkers Nijmegen, Netherlands More articles by this author , and Jack SchalkenJack Schalken Nijmegen, Netherlands More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.314AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Transient receptor potential vanilloid subtype 4 (TRPV4) is a non specific cation channel that is located on the epithelial cell membranes of the urinary bladder, the ureter and the distal section of the kidney tubuli. Transgenic TRPV4 deficient mouse have a phenotype that displays bladder dysfunction. Therefore, TRPV4 channels are being investigated as a mechanoreceptor in the bladder and as a potential pharmacological target for OAB. Besides this, numerous TRP-channels have recently been associated with carcinogenesis. Previous research has demonstrated a molecular connection between TRPV4 and barrier forming Adherence Junctions (AJ's) in all above mentioned tissues. Other groups have shown that the skin epithelium of TRPV4 deficient mice have an abnormal cell junction formation. The aim of this study is to investigate cell junction formation in the urogenital tract of humans and in TRPV4 knockout mice. METHODS The location of TRPV4, AJ's and Tight Junctions (TJ) was investigated with immunofluorescence assays, Western blotting and qPCR techniques on non cancerous tissue sections from human cystectomies (n=4), ureterectomies (n=1), nefrectomies (n=2) and kidney and bladder tissues from wild type and TRPV4 −/− mice. Subsequent evaluation of epithelial cell junctions (AJ and TJ) was investigated in bladder tissues from wild type and TRPV4 −/− mice with Transmission Electron Microscopy (TEM). RESULTS TRPV4 co localizes with adherence junctions throughout the urogenital tract (fig; top). Immunofluorescence assays demonstrated a qualitative and quantitative reduction of cell junction formation (both AJ and TJ) in TRPV4 −/− kidney and bladder epithelia (fig; bottom A,B). TEM evaluation confirmed these results and showed a remarkable increase in intercellular space between adjacent urothelial cells of the TRPV4 −/− mouse bladder (fig; bottom C). CONCLUSIONS TRPV4 channels are connected to epithelial adherence junctions throughout the urogenital tract and plays a role in cell junction formation. These result suggest that TRPV4 channels, besides being important for sensory functions, is also involved in epithelial barrier formation. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e104-e105 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Dick Janssen Nijmegen, Netherlands More articles by this author Kees Jansen Nijmegen, Netherlands More articles by this author John Heesakkers Nijmegen, Netherlands More articles by this author Jack Schalken Nijmegen, Netherlands More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

2,3,7,8-Tetrachlorodibenzo- p-dioxin induced autophagy in a bovine kidney cell line

The administration of 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD) to a variety of cultured cells may alter their ability to proliferate and die. In a previous study we demonstrated that TCDD induced proliferation in Madin-Darby Bovine Kidney (MDBK) cells where no signs of apoptosis were observed, but herein, analysis of MDBK cell morphology, in a large number of exposed cells, revealed some alterations, as expanded cytoplasm, an increase of intercellular spaces and many pyknotic nuclei. Hence, the aim of the current study was to elucidate the influences of dioxin on cell proliferation and cell death. We found that dioxin increased proliferation, as well as, activated cell death with autophagy, as we detected by increased amount of LC3-II, an autophagosome marker. Furthermore, formation of acidic vesicular organelles was observed by fluorescence microscopy following staining with the lysosomotropic agent acridine orange. These results were accompanied by down-regulation of telomerase activity, bTERT and c-Myc. Key tumor-suppressor protein p53 and expression of cell cycle inhibitor p21Waf1/Cip1 were activated after TCDD exposure. These changes occurred with activation of ATM phosphorylation in the presence of a decrease in Mdm2 protein levels. Taken together, these results support the idea that TCDD in MDBK cells, may exert its action, in part, by enhancing cell proliferation, but also by modulating the incidence of induced cell death with autophagy.

Alterações morfo-histológicas em larvas de Aedes aegypti (linnaeus, 1762) (Diptera, Culicidae) causadas pelo tanino catéquico isolado da planta do cerrado Magonia pubescens (Sapindaceae)

The control of Aedes aegypti is nowadays one of the major public health problemsin Brazil, being the main vector of dengue and urban yellow fever. Of Africanorigin, this mosquito is well adapted to Brazilian climatic conditions and is nowdistributed in all states. The usual methods to combat and control the mosquito havebeen highly questioned. The chemical insecticides currently used have led to theemergence of resistant populations of mosquitoes and the consequent increase ofits density, causing significant epidemics of dengue. Insecticides of botanical originhave shown to be alternative candidates to their control. This work presents forthe first time, morphohistologic changes caused by the larvicidal activity of tannincatechin extracted from Magonia pubescens on larvae of A. aegypti, showing thepotential of this natural larvicidal compound. Larvae of the 3rd stage were submittedto a water solution of tannin catechins, at 37ppm, where they remained for up to 24h. Larvae that reached lethargic state were collected and fixed in paraformaldehydeat 4%, in sodium cacodylate buffer at 0.1 M and pH 7.2, included in resin, mountedon slides, stained with hematoxylin-eosin and analyzed by light microscope. Themain cell toxic effects of tannin catechin on larvae of A. aegypti were the highvacuolation and absence of cytoplasmic limits, vesicular apical formation withrelease of cytoplasmic content, increase in intercellular spaces and detachment ofcells from the basal membrane.

Impact of metals on histopathology and expression of HSP 70 in different tissues of Milk fish ( Chanos chanos) of Kaattuppalli Island, South East Coast, India

Histological and Immunohistochemical studies were carried out to document the possible impact of heavy metal contamination in different tissues of Chanos chanos. Heavy metals such as Cu, Pb, Zn, Cd, Mn and Fe were predominant in water, sediment and biota of Kaattuppalli Island and varied significantly between two different sites. Histological changes such as swelling of muscle fiber and break down of muscle bundles were noted in the muscle. Similarly gill filament cell proliferation, increase in intercellular spaces and primary and secondary lamellar epithelium were evident in gills. The hepatocytes showed damage of central vein and rupture of irregular hepatic plate with more number of vacuoles in the fish collected from polluted site compared to that from the less polluted site. The impact of pollution was also assessed in different tissues by immunohistochemistry using primary antibody (mouse monoclonal HSP70 antibody 1:2000) and secondary antibody (HRP conjugated antibody) for expression of stress protein. Immunostaining analysis showed expression of HSP70 with high intensity in the tissues of fish collected from polluted site compared to less polluted sites. Further, HSP70 positive cells were analyzed from six locations per fish tissue section. One-way analysis of variance (ANOVA) followed by least significant difference (LSD) was used to check if the expression was significant. Results indicate that the values are statistically significant at the two different sampling sites ( P < 0.05) .

Pathological evaluation of pancreatic exocrine glands in experimental fluorosis

Objective To monitor the pathological alterations in pancreas of rat during experimental fluorosis. Methods Sixty Sprague Dawley albino rats of both sexes were divided into 12 experimental groups and one control group. The rats of control group were administered subcutaneously double distilled water 1 mL/kg bw daily. The experimental groups were injected with 30, 45, and 75 mg NaF/kg bw/day. The experimental period was divided into 4 phases at interval of 15, 22, 30, and 36 days. Animals were sacrificed from each group at the end of 16, 23, 31, and 37 days. Results The following changes were observed in this study: (1) Pathological examination of pancreas after 15 days of fluoride treatment revealed: hypertrophy of acini, leucocytes infilteration and pycnotic nuclei due to necrosis of acini in group 1; uremic alterations, invulsion and infoldings of reticular layer of islets of Langerhans in group 2; and a decrease in number of acini and interlobular connective tissues resulted in an increase in intercellular spaces in pancreas of rats in group 3. (2)Hyalinization and hypertrophy in the lobules of acini and hyperplasia and hypertrophy in intercalated duct with mucinous secretion in pancreas of rat of group 4; squamous metaplasia of pancreatic duct, adenoma of pancreas, hemorrhagic necrosis in group 5; and hyperplasia of acini and reduction in number of pancreatic islets in group 6. (3) Disorganization and atrophy of pancreatic lobules and presence of vacuoles in a group of six were visible in pancreas of rats in group 7; acute pancreatic and lamellated inflammatory cells in test rats of group 8; and islet adenoma and decrease in number of islets cells, and exudation in acini were noticed in experimental rats of group 9. (4) In the last phase of experimentation, atrophic alterations in pancreatic acini, invulsions, and necrosis was prominent in group 10, deep inflammation and proliferation of connective tissue of pancreas in experimental group 11, and periodical fibrosis, hyperplasia of acini, degenerative changes in pancreas of rats in group 12. Conclusion The histopathological examination of pancreas of fluoridated rats exhibited structural alterations in the exocrine glands. The acini revealed hypertrophy, pyknotic nucleus, necrosis and uremic alterations. Acini became lobulated and reveal increased pigmentation.