DigitalUM :: Browsing by Subject "Ethanol"

Browsing by Subject "Ethanol"

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Open Access
Assessing DESS solution for the long-term preservation of nematodes from faecal samples
(Elsevier, 2022-10-18) Ruiz de Ybáñez Carnero, María del Rocío; Rodríguez-Caro, R.C.; Maíz-García, A.; Gómez, L; Giménez Casalduero, Andrés; Graciá, E.; Gonzálvez Juan, Moisés; Sanidad Animal
Preservation of biological samples is a relevant issue for many scientific disciplines. Although traditional preservers, such as formaldehyde or ethanol, imply major disadvantages related to health risks, DNA degradation and distortion of structures, they are widely used. Hence, the search for viable alternatives preserving morphometry and genetics seems necessary. Here we assess the suitability of DESS solution to preserve adult nematodes and their eggs in faeces. Concretely, faecal samples of terrestrial tortoises with oxyurids were used to: (i) compare the 1-month storage efficacy of eggs from different conservation protocols (faeces without preserver at -20 °C, faeces with DESS solution at room temperature, faeces with DESS solution at -20 °C and faeces with ethanol 70% at room temperature); (ii) address morphological nematode identification after 2 years of storage with DESS. We also corroborated that nematode DNA remained viable after 2 years. Overall, our results showed that DESS solution at room temperature is an advisable alternative to conserve both parasite eggs and adult nematodes for morphological identification and genetic purposes. It also offers the advantages of being low-cost, safe and suitable for fieldwork conditions and shipments without refrigeration for nematode preservation.
Open Access
Chronic ethanol feeding alters the epithelial cell proliferation and apoptosis in rat gastric mucosa
(Murcia : F. Hernández, 2007) Ge, Y. B.; Du, J.; Fan, L.L.; Li, Y.C.; Gu, L.
We developed a chronic drinking rat model to investigate the long-term effects of ethanol feeding on cell proliferation and apoptosis in rat stomach. Adult male Sprague-Dawley (SD) rats received either an isocaloric control or drinking water containing 6% (v/v) ethanol as their only water intake for 1, 3, 7, 14 and 28 days. At the end of each feeding period, animals were sacrificed and the stomach was dissected for the sample preparation. The cell proliferation and apoptosis in gastric mucosa of rats in different groups were analyzed by flow cytometer, immunohistochemistry and computer image analysis. In the flow cytometric study, compared with the control, the cell apoptosis in gastric mucosa of the rats was enhanced during the exposure to the ethanol in 3rd to 28th day. Otherwise the cell proliferation was increased in 3rd to 14th days, and decreased in 28th days, respectively. The results were confirmed by immunohistochemistry and computer image analysis studied. This finding suggested that short-term chronic adequate alcohol intake may enhance the cell turnover of gastric mucosa. Long-term stimulus with the low concentration ethanol may cause the impairment of the cell turnover function of the gastric mucosa and may be one of the mechanisms underlying the gastric pathology associated with alcohol abuse.
Open Access
Effect of ethanol on the cerebellar cortex of the chick embryo
(Murcia : F. Hernández, 1990) Quesada, A.; Prada, F. A.; Espinar, A.; Genís-Gálvez, J. M.
Open Access
Effects of chronic administration of either ethanol or pentanol on rat duodenum morphology
(Murcia : F. Hernández, 2002) Vaquera, J.; Vaquera, A.; Girbes, T.
The morphology of the rat duodenum after chronic treatment with 15% (v/v) ethanol and 4% (v/v) pentanol was studied. Male Wistar rats of experimental groups were given ethanol and pentanol for 15 weeks with food and fluid freely available. Ethanol-15% and 4% pentanol-fed rats showed a significantly reduced fluid and food intake as compared with control rats. The study of the mucosa indicated that the number of chronic inflammatory infiltrating (mononuclear cells) and goblet cells was higher in the groups of the ethanol- and pentanol-fed rats than in the control group. There was an increase in the thickness of the brush border in pentanolfed rats. Intervillus adhesion was concurrently observed in the pentanol-fed rats but not in the control or ethanolfed rats. After ethanol feeding many of the villi developed blebs at the apex of the villus or laterally on its upper half. These blebs generally remained intact. In contrast, after pentanol feeding no bleb formation was appreciated. The intake of ethanol and other short chain alcohols present in alcoholic beverages leads to mainfold disturbances on the rat duodenum. These findings suggest that the chronic ingestion of pentanol seems to promote cellular changes but less important than those observed after chronic ethanol ingestion.
Open Access
Effects of chronic ethanol administration on the serotonin-producing cells in rat gastric antral and duodenal mucosa
(Murcia : F. Hernández, 1993) Todorovic, V.; Koko, V.; Varagic, J.; Lackovic, V.; Vuzevski, V.D.; Milin, J.
The present study describes our observations on optical and ultrastructural features of serotonincontaining cells in the rat antral and upper duodenal mucosa, utilizing optic morphometric measurements in a model of experimental chronic alcoholism of rat in which nutrition was well controlled. Male Wistar rats were given ethanol to provide 23 per cent of the total calories, while starch replaced ethanol isocalorically in controls. Twenty-five per cent of the calories were provided by protein in both groups. Blood levels of serotonin were significantly raised after chronic ethanol feeding (0.05910.06 vs. 0.15910.012 pglml, p<0.01). Decrease in the number of immunohistochemicallydetectable serotonin-containing cells was found in the pyloric gland mucosal area specimens of the chronically ethanol-treated rats (68.915.2 vs 43.313.0; p<0.001). The immunohistologically-evaluated number of the same cells in the duodenal mucosa specimens was significantly decreased by alcohol feeding. Although total villi and crypt count per whole circular section, and the number of crypts per villus were not significantly changed either in control animals or in chronically ethanol-fed rats, decreased number of these cells per whole circular section (289121.6 vs. 183f 10.5; p
Open Access
Effects of ethanol on the ultrastructure of the hamster femur
(Murcia : F. Hernández, 2001) chen, H.; Hayakawa, D.; Emura, S.; Ozawa, Y.; Taguchi, H.; Yano, R.; Shoumura, S.
Several previous studies have indicated that chronic ingestion of ethanol exerts harmful effects on bones. However, few data are available concerning the effects of ethanol on the ultrastructure of bone. To further elucidate the effects of ethanol on bone, we studied the morphology of femur in golden hamsters after long-term treatment with ethanol. Six-week-old male hamsters were divided into 4 groups. Ethanoltreated animals were given ethanol at a concentration of 7% with food and water freely available, whereas the pair-fed animals (weight-matched to ethanol hamsters) had tap water available as the only drinking fluid. The femur weight, blood ethanol and serum calcium concentrations were determined after 3 and 5 months. The bone mineral density (BMD) of the whole body was measured before and after the experiment. Femurs of both sides were dissected and processed for morphometric measurement, light microscopy, scanning and transmission electron microscopy. In the ethanoltreated hamsters, BMD of the whole body and the weight of femur tended to decrease when compared with those of the controls. Light microscopy and scanning electron microscopy showed that the trabecula in the dista1 end of the femur from ethanol-treated hamsters were thinner than those of the controls. We also observed the disrupted swollen mitochondria of the femoral osteoblasts and osteocytes in the ethanol-treated hamsters. No significant difference in serum calcium levels and femoral osteoclasts was found. These results indicate that long-term treatment with ethanol results in disruption of femoral osteoblasts and reduction of bone mass in trabecular bone.
Open Access
Enhanced autophagy and phagocytosis of apoptotic lymphocytes in splenic macrophages of acute ethanol-treated rats: Light and electron microscopic studies
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2024) Betsuyaku, Tsubasa; Ito, Yuko; Peake, Nicholas; Al Bari, Abdul Alim; El Akabawy, Gehan; Eid, Nabil
Autophagy is a prosurvival mechanism for the clearance of damaged cellular components, specifically upon exposure to various stressors. In lymphoid organs, excessive ethanol consumption increases lymphocyte apoptosis, resulting in immunosuppression. However, ethanol-induced autophagy and related phagocytosis of apoptotic lymphocytes in the spleen have not been studied yet. Adult male Wistar rats were injected intraperitoneally either with 5 g/kg ethanol or phosphate-buffered saline (as a control group) and then sacrificed 0, 3, 6, and 24 hours after injection. Light and transmission electron microscopy (TEM) findings indicated enhanced T cell apoptosis in the white pulps of ethanol-treated rats (ETRs) compared with the control group, which peaked at 6h and was associated with the accumulation of tingible body macrophages (TBMs). These macrophages exhibited an upregulated autophagic response, as evidenced by enhanced LC3-II (a specific marker of autophagosomes) expression, which peaked at 24h. In addition, double labeling immunofluorescence of LC3-II with lysosomal markers revealed the enhanced formation of autolysosomes in TBMs of ETRs, which was associated with suppression of p62 immunostaining, indicating the enhanced autophagic flux. Interestingly, this elevated autophagic response in ETR TBMs was accompanied by evidence of LC3-associated phagocytosis (LAP) of apoptotic splenocytes. This is based on TUNEL/LC3-II double labeling and TEM observations of phagosomes containing apoptotic bodies, enclosed within phagosomal membranes adjacent to the autophagic vacuoles. It can be concluded that enhanced prosurvival autophagy in splenic TBMs of ETRs and clearing of apoptotic lymphocytes via LAP may contribute to preventing secondary necrosis and autoimmune diseases.
Open Access
Ethanol enhances thymocyte apoptosis and autophagy in macrophages of rat thymi
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Betsuyaku, Tsubasa; Eid, Nabil; Ito, Yuko; Tanaka, Yoshihisa; Otsuki, Yoshinori; Kondo, Yoichi
Tingible body macrophages (TBMs) play essential roles in the phagocytosis of apoptotic lymphocytes, specifically under exposure to various stressors. Although excessive ethanol consumption may enhance thymocyte apoptosis, reports investigating the autophagic response of the thymus to ethanol toxicity are still lacking. We investigated apoptosis and autophagy in thymi of an animal model of binge ethanol exposure. Adult male Wistar rats were injected intraperitoneally either with 5 g/kg ethanol or phosphate buffer saline (for the control group) and sacrificed 0, 3, 6 and 24 hours after injection. Light and transmission electron microscopy (TEM) studies revealed enhanced formation of TBMs phagocytosing many apoptotic thymocytes in the thymic cortex of the ethanol-treated rats (ETRs), and this formation was particularly marked at 24 h. The macrophages showed signs of activation under TEM and immunofluorescence double labeling with RM4 (a macrophage marker) and iNOS. Additionally, in comparison to the control group, autophagy was enhanced in ETR thymic TBMs as evidenced ultrastructurally by accumulation of autophagic vacuoles, immunohistochemical increases in LC3 puncta, Western blot analysis of the latter protein, and colocalization of LC3 and RM4 in immunofluorescence double labeling. Immunoelectron microscopy also revealed LC3-labeled autophagic vacuoles and apoptotic cell phagosomes in ETR TBMs, suggesting the possibility of LC3-related phagocytosis. This was confirmed by enhanced colocalization of LC3 with lysosomal cathepsins in double labeling. These results indicate that enhanced autophagy in ETR thymic TBMs is not only a cytoprotective mechanism but could also be involved in the clearance of apoptotic thymocytes, thus preventing autoimmune reactions and suppressing inflammatory response.
Open Access
Ethanol-induced mitophagy in liver is associated with activation of the PINK1-Parkin pathway triggered by oxidative DNA damage
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Eid, Nabil; Ito, Yuko; Horibe, Akio; Otsuki, Yoshinori
Mitophagy is a cytoprotective mechanism against mitochondrial damaging agents. Studies demonstrating morphological evidence for the involvement of the PINK1-Parkin pathway in the hepatocyte mitophagic response to ethanol toxicity, and potential links to apoptosis and mitochondrial alterations such as spheroid formation are still lacking. We addressed these unresolved issues using a rat model of binge alcohol exposure. Adult rats were injected with ethanol (5g/kg) and liver samples were taken at 0, 3, 6, and 24 hours after ethanol administration and processed for light and electron microscopic studies. Ethanol induced a low level of hepatocyte apoptosis, peaking at 3 h and decreasing significantly by 24 h. In contrast, there was enhanced formation of mitophagic vacuoles in the majority of normal hepatocytes of ethanol-treated rats (ETRs), which peaked at 6 h and was maintained up to 24 h based on electron microscopy and TUNEL/LC3 double labelling. Moreover, enhanced mitophagy in ETR hepatocytes was confirmed by increased LC3 puncta formation, and co-localization of Parkin and LC3 with mitochondrial and lysosomal markers. Immunoelectron microscopy demonstrated the localization of PINK1 and Parkin to damaged mitochondria of ETR hepatocytes, which was consistent with co-localization of Parkin with 8-OHdG, a marker of oxidative mitochondrial DNA damage. Furthermore, electron microscopy showed enhanced formation of mitochondrial spheroids in ETR hepatocytes. These data are the first direct morphological evidence linking PINK1-Parkin pathway activation to the enhanced mitophagic response of hepatocytes to ethanol toxicity. Ethanol-induced hepatic mitophagy may be a prosurvival mechanism, which may have therapeutic implications.
Open Access
Microscopic correlates of adaptive cytoprotection in an ethanol injury model
(Murcia : F. Hernández, 1989) Schmidt, Carmen L.; Smith, Gregory S.; Miller, Thomas A.
The present study histologically investigated the efficacy of pretreating rat gastric mucosa with the mild irritants, 10% and 25% ethanol (EtOH), against the known damaging effects of 100% EtOH. Fasted rats received 1 m1 of either water, 10% EtOH, or 25% EtOH by orogastric intubation. Fifteen minutes later, a portion of these animals was sacrificed and tissue samples of the oxyntic region of the stomach were excised and processed for quantitative histologic analysis. Remaining animals received a 1 m1 oral bolus of the necrotizing agent, 100% EtOH. Five minutes later, these animals were sacrificed and tissues were prepared in a like manner. In a separate series of experiments, the aforementioned protocols were repeated, except that al1 animals received the prostaglandin synthetase inhibitor, indomethacin (5.0 mg/kg intraperitoneally), 30 min before administration of the mild irritant. Microscopically, the administration of water or 10% EtOH alone caused a small and comparable amount of superficial injury to the gastric mucosa. Moreover, both substances failed to induce protection in stomachs subseqently exposed to 100% EtOH. Indomethacin pretreatment did not significantly alter any of these findings. In marked contrast, 25% EtOH alone elicited a substantial degree of superficial damage to the gastric mucosa. Nevertheless it significantly reduced the depth of injury in animals subsequently challenged by 100% EtOH. Indomethacin failed to aggravate the effects of 25% EtOH alone, but partially inhibited the protective effect of this mild irritant against 100% EtOH induced damage. Our findings indicate that adaptive cytoprotection is a real phenomenon that can be demonstrated microscopically. Such protection is limited primarily to the deep mucosa1 layers (i.e. gastric glands), appears in part to be prostaglandin mediated and seems to require the generation of moderate surface cell damage (as occurred with 25% EtOH, but not 10% EtOH) to induce its initiation.
Open Access
Modificaciones estructurales y ultraestructurales en la mucosa gastrointestinal por acción de radicales libres. Posible efecto protector del etanol
(Asociación Española de Toxicología, 2001) Gómez Zapata, M.; Vicente Ortega, V.; Martínez Díaz, F.; Ordóñez Escudero, D.; Luna Maldonado, A.; Falcón Romero, María; Oftalmología, Optometría, Otorrinolaringología y Anatomía Patológica
El objetivo de este trabajo es estudiar como se modifica la estructura de la mucosa gastrointestinal al verse expuesta a radicales libres y determinar si el etanol puede disminuir los daños producidos por estos radicales libres. Para ello hemos estudiado las alteraciones histológicas (estructurales y ultraestructurales) de la mucosa gastroduodenal de ratas tratadas con el reactivo de Fenton (generador de radicales libres) con y sin etanol. Nuestros resultados muestran que a nivel estructural, el grupo de ratas a las que se administró etanol junto con el reactivo de Fenton presentaban menos lesiones de tipo inflamatorio que el grupo de animales tratados solo con el reactivo de Fenton sin etanol, lo que indica que el etanol sí amortigua la acción de estos radicales libres. Sin embargo a nivel ultraestructural no podemos hacer diferenciaciones entre los dos grupos ya que todas las lesiones encontradas son lesiones celulares inespecíficas y aparecen con igual frecuencia en los mismos. Por lo tanto, debido a la inespecificidad de las alteraciones histológicas producidas por los radicales libres, estas solo nos pueden reflejar la intensidad del daño producido pero no su etiología.------------

Browsing by Subject "Ethanol"

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