Amount Of Mucus Research Articles

Inhaled Simvastatin Nanoparticles for Inflammatory Lung Disease

Current inhaled treatments are not adequate to treat all lung diseases. In this study, a promising nanotechnology has been developed to deliver a potential anti-inflammatory and muco-inhibitory compound, simvastatin, for treatment of inflammatory lung diseases via inhalation. Simvastatin nanoparticles (SV-NPs) encapsulated with poly(lactic-co-glycolic) acid were fabricated using the solvent and anti-solvent precipitation method. SV-NPs were found to be stable up to 9months at 4°C in a freeze-dried form prior to reconstitution. The amount of mucus produced was significantly reduced after SV-NPs treatment on inflammation epithelial cell models and were effective in suppressing the proinflammatory marker expression. This study suggests that SV-NPs nebulization could potentially be used for the treatment of chronic pulmonary diseases.

CON4EI: Slug Mucosal Irritation (SMI) test method for hazard identification and labelling of serious eye damaging and eye irritating chemicals

Assessment of ocular irritancy is an international regulatory requirement in the safety evaluation of industrial and consumer products. Although many in vitro ocular irritation assays exist, alone they are incapable of fully categorizing chemicals. The objective of CEFIC-LRI-AIMT6-VITO CON4EI (CONsortium for in vitro Eye Irritation testing strategy) project was to develop tiered testing strategies for eye irritation assessment that can lead to complete replacement of the in vivo Draize rabbit eye test (OECD TG 405). A set of 80 reference chemicals was tested with seven test methods, one method was the Slug Mucosal Irritation (SMI) test method. The method measures the amount of mucus produced (MP) during a single 1-hour contact with a 1% and 10% dilution of the chemical. Based on the total MP, a classification (Cat 1, Cat 2, or No Cat) is predicted. The SMI test method correctly identified 65.8% of the Cat 1 chemicals with a specificity of 90.5% (low over-prediction rate for in vivo Cat 2 and No Cat chemicals). Mispredictions were predominantly unidirectional towards lower classifications with 26.7% of the liquids and 40% of the solids being underpredicted. In general, the performance was better for liquids than for solids with respectively 76.5% vs 57.1% (Cat 1), 61.5% vs 50% (Cat 2), and 87.5% vs 85.7% (No Cat) being identified correctly.

Uretrolitíase obstrutiva em equino

Background: The presence of macroscopic concretions in the urinary system is called urolithiasis. In horses the occurrence is rare. The calculi are generally large and unique spicule concretions, composed of calcium carbonate crystals. Less commonly, the concretions are of mixed crystals of calcium carbonate and of calcium phosphate. The aim of the this work was to report a case of obstructive urethrolithiasis in a horse.Case: A 1-year-and-5-month-old, undefined breed, uncastrated male presented tenesmus, strangury, and dysuria. The horse died seven days after the onset of clinical signs and was referred for necropsy. At necropsy, samples were collected from organs of the abdominal cavity, thoracic cavity, central nervous system and integumentary system. The fragments were fixed in 10% formaldehyde, cleaved, routinely processed for histology and stained with hematoxylin and eosin. A urinary calculus collected during necropsy was sent for chemical analysis. In the abdominal cavity, a large quantity of citrus liquid of uremic odor was observed. Deposition of fibrin filaments in the peritoneum was observed, as well as in diaphragm and abdominal organs. Serosa of the small and large intestine was red with evidence of the capillaries. Kidneys presented multiple slightly protrusive white areas with a maximum diameter of 3 mm. When cutting, these areas were limited to the cortical region. There was rupture of the urinary bladder, diffuse loss of the mucosa with capillary exposure and evidence, as well as the deposition of fibrin and crude blood clots. The opening of the penile urethra presented a urolith of approximately 3.5 x 3 cm in size, with a rough and porous surface, light brown and of a stony consistency. Necrosis of the mucosa and hemorrhage were also observed in the urethra. Microscopically, the kidney showed multiple foci formed by intense inflammatory infiltrate composed of macrophages, neutrophils and plasma cells in the interstitium of the cortical region. In the urinary bladder, there was diffuse necrosis of the lining epithelium, marked inflammatory infiltrate composed mainly of neutrophils in the submucosa, in addition to fibrin deposition and edema. Penile urethra presented changes similar to those described in the urinary bladder. The chemical analysis of the urinary calculus demonstrated the presence of calcium oxalate, triple magnesium ammonia (struvite) phosphate and ammonium in its composition.Discussion: The diagnosis of obstructive urethrolithiasis was established from the clinical-pathological condition. The occurrence of urolithiasis in horses is rare. The lower occurrence in this species is probably attributed to the large amount of mucus produced by the glands in the renal pelvis and the proximal ureter, which acts as a lubricant to prevent the adhesion of the crystals to the urothelium. The absence of sigmoid flexure and vermiform urethral process in equine also influence the low occurrence of the disease, when compared to ruminants. Rupture of the urinary bladder followed by uroperitonitis and nephritis as observed in this case are common consequences in obstructive urolithiasis. Uroliths in equines are generally composed of a variety of hydrated calcium salts and calcium carbonate, magnesium, and phosphorus salts, with less common struvite compounds. Urethrolithiasis is poorly described in horses and can be caused by struvite stones.

The gastroprotective effect of ethyl acetate fraction of hot water extract of Trichosanthes cucumerina Linn and its underlying mechanisms

BackgroundAntacids, anticholinergic drugs, histamine H2- receptor antagonists and irreversible proton pump inhibitors have been used for the treatment of gastric ulcers. However, prolonged use of these drugs may lead to series of adverse effects such as diarrhea, headache, rash, hypertension, muscular and joint pain. Therefore, there is an urgent need of more effective and safer treatments with fewer side effects. The aim of the present study was to scientifically evaluate the gastroprotective activity of fractions of the hot water extract of Trichosanthes cucumerina Linn (Family: Cucurbitaceae) aerial parts with a view to identifying the fraction with the best gastroprotective activity and the possible mechanism/s by which this fraction exert gastroprotection.MethodsGastroprotective activity of hexane fraction (HF), ethyl acetate fraction (EF), butanol fraction (BF) and aqueous fraction (AF) were evaluated by the assessment of ability to reduce the ulcer index in ethanol-induced rat model and the mode of action by which the most active fraction mediating gastroprotection.ResultsEF showed the maximum gastroprotection effect followed by BF and AF. EF (75 mg/kg) exhibited significantly higher gastroprotection compared to the reference drugs. Further investigations with two lower doses of EF confirmed that EF can mediated a significant and dose dependent gastroprotection. The rats treated with the EF showed significant reduction in free acidity (45%), total acidity (by 48%) in the gastric juice, increased the amount of mucus produced by the rat gastro mucosa and potent antihistamine activity (by 25.6%). EF was also rich in phenolic compounds and flavonoids.ConclusionGastroprotective mechanism of EF is possibly involves inhibition of acidity, elevation in mucus content, inhibition of histamine and antioxidant mechanisms.

Ultramorphological changes in gill rakers of Astyanax altiparanae (Characidae) kept in contaminated environments.

Most water bodies in Brazil, and in the world, are contaminated by some types of pollutants, ranging from sewage to metal/chemicals, carcinogenic products, and biodegradable detergents. Despite the extensive knowledge on their effects on fish biology and especially on gill morphology, research that concerns their impacts on gill rakers and implications in parameters such as food consumption cannot be found in the literature. Gill rakers are vital because, together with gills, they are responsible for the defense and protection of the organism and for selecting appropriate food for survival. When detergents, which can act as toxic chemical agents, get in contact with the body of the fish, they can cause severe effects that must be understood. Therefore, our study investigated ultramorphological changes in gill rakers of Astyanax altiparanae (Lambeth) caused by the exposure to biodegradable detergents. Fish were exposed to a 1mg/L dilution of a mixture of detergents and pure water from an artesian well for 5months. Results revealed that the first month of exposure to detergent caused dilation of chemical receptors in taste buds and the rise of a large number of orifices for mucus release among pavement cells in gill rakers, although only a small amount of mucus was found in fish exposed both to pure water and the detergent dilution. After 5months, there was an increase in the dilation of these chemoreceptors, excess mucus on gill rakers of detergent groups, and the emergence of microbridges between microridges in pavement cells.

Optimization of the Isolation Procedure of Genomic DNA from a Mucus Laden Pulmonate Gastropod, Achatina fulica

Genomic DNA of good quality is a prerequisite in molecular biology studies involved in deciphering the information contained in the molecules that are chosen for studying the evolutionary interrelationships among taxa. Genomic DNA should be free of contaminants which may otherwise interfere while performing PCR. This is particularly essential while isolating genomic DNA from gastropods due to the presence of large amounts of mucopolysaccharides in their tissues. In the present study, we have developed a new method for obtaining high quality genomic DNA from a pulmonate snail Achatina fulica, which secretes a large amount of mucus. In this method, we have obtained good quality genomic DNA, compatible for downstream molecular biology applications like PCR and sequencing of 16S rDNA. The yield of the genomic DNA was 2790 µg/ml. This method may be extended for isolating high quality genomic DNA from different gastropods native to Indian sub-continent, which serves as a starting point for studying their phylogenetic affinities.

Effect of soaking seeds of flax on the ultimate strength of flaxes

There are one of ways to save peoples heath is eating food, what rich in fiber. To the recovery of polyunsarurated fatty acids, protein, mineral substances, fibers it recommended to eat flax seeds and its products. One of these products are flaxes. The purpose of the work was a rational choice of the duration of soaking seeds, providing strong reception of finished products. Founds that when hydrated of 10 to 30 minutes of the study setting almost unchanged. After 30 minutes soaking the tensile strength increased, and then decreased. The maximum value observed in the samples in which lasted 40 minutes soaking. Probably, due to the fact that upon contact with water 30 minutes before the moisture is adsorbed on the surface of flax seed are thus formed in a small amount of mucus. From 30 to 40 minutes soaking carbohydrates undergo hydrolysis inner layers of the endosperm and seed shell. While the water absorption capacity of flax reaches limits. Also increases the amount of mucus, and after 40 minutes soaking becomes excessive, resulting in reduced tensile strength of the finished product. Therefore, rational while soaking flax 40 minutes, providing the maximum value of the parameter under study. Flux thus, thanks to the ability of flax seed soaking and store slime forming after drying alpha-linolenic acid is a source of polyunsaturated fatty acids, dietary fibers, vitamins, minerals.

Clinicopathologic analysis of low grade appendiceal mucinous neoplasm

The clinicohistological features of 12 patients with low grade appendiceal mucinous neoplasm admitted from July 2011 to July 2015 were retrospectively analyzed. Ten patients complained lower abdominal pain, 5 cases presented space occupying lesions or abscess in appendix with ultrasound or CT examination, 2 cases presented cystic space occupying lesions in ileocecus on medical examination. The patients were generally characterized by enlargement of appendix, and cystic formation filled with jelly material. Histopathology showed appendix cavity expansion, lining single-layer mucus columnar epithelium with mild atypia and local papillary hyperpasia; appendix cavity partly bursted, and a large amount of mucus filled with appendix wall. Calcifications were found in 6 cases, floating broken epithelioid cells in 2 cases, calcified schistosome eggs in 2 cases. It is significant to understand the clinical and pathological features of LAMN. Key words: Appendiceal neoplasms; Mucus; Clinicopathologic features

Gastroprotective activity of the rhizome ethanol extract of Zingiber simaoense Y. Y. Qian in rats

Ethnopharmacological relevanceZingiber simaoense Y. Y. Qian belongs to the Zingiberaceae family. Its rhizome has been used in Thai folk medicine to relieve gastric disorders; however, scientific evidence of its pharmacological activities has not yet been revealed. Aim of the studyThis study was designed to validate the gastroprotective activity and to identify possible mechanisms of gastroprotection of Z. simaoense rhizome ethanol extract (ZSE) in rats. Materials and methodsThe gastroprotective effect of ZSE was tested using models of gastric ulcers induced by acidified ethanol, indomethacin, and restraint water immersion stress. Models for determination of gastric wall mucus secretion and plasma malondialdehyde levels as well as pylorus ligation were used to explore the mechanisms of action. ResultsAfter oral administration by intragastric gavage, ZSE 7.5, 15, and 30mg/kg or cimetidine 100mg/kg significantly inhibited the formation of gastric ulcer in all gastric ulcer models. The gastric wall mucus amount was significantly higher than that of the ulcer control group, plasma malondialdehyde levels were normalized, and gastric secretion was partly inhibited by pretreatment with ZSE. ConclusionsThis study demonstrates the gastroprotective activity of ZSE in rats. The mechanisms of action of ZSE may depend on its ability to maintain the integrity of gastric wall mucus through the protection of gastric mucus, and/or by increasing the gastric mucus synthesis and secretion through prostaglandin synthesis. Moreover, the antioxidant activity of ZSE may also contribute to its mechanism of gastroprotection.

Pyunkang-hwan (Pyunkang-tang) ameliorates air pollutant-induced inflammatory hypersecretion of airway mucus and bleomycin-induced pulmonary fibrosis in rats

OBJECTIVETo investigate the effect of Pyunkang-hwan (Pyunkang-tang) extract (PGT) on secretion of airway mucin in an experimental animal model involving hyperplasia of goblet cells and mucus hypersecretion, and to test its effects on bleomycin (BLM)-induced pulmonary fibrosis in vivo. METHODSThe protective activity of orally administered PGT was assessed in two rat pulmonary disease models. Effects on hypersecretion of pulmonary mucin in sulfur dioxide (SO2)-induced bronchitis in rats were assessed by quantifying the amount of mucus secreted and examining histopathology in the tracheal epithelium. In a rat model for BLM-induced pulmonary fibrosis, toxicity to the pulmonary system was examined by measuring levels of malondialdehyde and hydroxyproline, indicators of lipid peroxides and collagen, respectively, in lung tissue 28 days post-BLM treatment. Serial sections of lung tissue were stained with Masson trichrome to visualize collagen deposition. Effects of PGT on collagen synthesis were also assessed in vitro, in a cell culture model. RESULTSPGT inhibited mucin secretion and normalized SO2-induced increased mucosubstances in goblet cells. In the BLM-induced model, PGT decreased the characteristic histopathological features of lung fibrosis and inhibited fibrotic lesions, as indicated by decreased hydroxyproline content. PGT also inhibited the BLM-induced increase in malondialdehyde levels, demonstrating its protective effect against lipid peroxidation in cell membranes of the lung. In MLg 2908 mouse lung fibroblast cells, PGT decreased transforming growth factor (TGF)-β-stimulated type I collagen synthesis. CONCLUSIONSPGT can inhibit both hypersecretion of airway mucins and pulmonary fibrosis.

Can gelatinous zooplankton influence the fate of crude oil in marine environments?

Gelatinous zooplankton are known for their capacity to excrete copious amounts of mucus that can be utilized by other organisms. The release of mucus is exacerbated by stressful conditions. Despite the recognized importance of cnidarian mucus to production and material flux in marine ecosystems, the role of gelatinous zooplankton in influencing the fate of oil spills is unknown. In this study we used laboratory experiments to observe the influence of mucus from the moon jellyfish (Aurelia aurita) on the aggregation and degradation of crude oil. The results show that jellyfish swimming in a dispersed solution of oil droplets produced copious amounts of mucus and the mucus aggregates that were shed by the animals contained 26 times more oil than the surrounding water. Incubation experiments showed that hydrocarbon degrading bacteria cell densities more than doubled in the presence of mucus and after 14days, resulted in a significant increase in oil degradation. These results suggest that jellyfish can aggregate dispersed oil droplets and embed them within a matrix that favors hydrocarbon degrading bacteria. While this study lends support to the hypothesis that the presence of gelatinous zooplankton can impact oil spills large scale mesocosm studies will be needed to fully quantify the influence on a natural system.

Development of a physiologically relevant dripping analytical method using simulated nasal mucus for nasal spray formulation analysis

Current methods for nasal spray formulations have been elementary evaluating the dripping characteristics of a formulation and have not assessed the behavior of the nasal formulation in the presence of varying types of mucus depending on the indication or diseased state. This research investigated the effects of nasal mucus on the dripping behavior of nasal formulations and focused on developing an improved in vitro analytical test method that is more physiologically relevant in characterizing nasal formulation dripping behavior. Method development was performed using simulated nasal mucus preparations for both healthy and diseased states as coatings for the dripping experiment representing a wide range of viscosity. Factors evaluated during development of this in vitro test method included amount of mucus, application of mucus, drying times, and compatibility of the mucus on a C18 Thin Layer Chromatography (TLC) substrate. The dripping behavior of nasal formulations containing a range of 1% Avicel to 3.5% Avicel was assessed by actuating the nasal spray on a perpendicular TLC plate coated with either healthy or diseased simulated nasal mucus. After actuation of the nasal spray, the dripping of the formulation on the coated TLC plate was measured after the plate was repositioned vertically. The method that was developed generated reproducible results on the dripping behavior of nasal formulations and provided critical information about the compatibility of the formulation with the nasal mucus for different diseased states, aiding in nasal spray formulation development and physical characterization of the nasal spray.

Mechanisms involved in pearlfish resistance to holothuroid toxins

Holothuroids produce triterpenoid saponins that act as chemical defenses against predators and parasites. These saponins interact with sterols of the plasma membranes, inducing the formation of pores and then cell lysis. To avoid such harms from their own saponins, holothuroids present specific sterols in their tissues. Despite the noxious cytotoxic effect of their chemical defenses, holothuroids host various associates that display specific adaptations to resist to saponin toxicity. Among them, symbiotic carapid fishes (i.e., pearlfishes) are resistant to ichthyotoxic saponins as they display no stress response and a survival time 45 times longer than free-living fishes without any specific gill adaptation. The present study aims at discovering the resistance mechanism(s) developed by carapids by addressing 3 hypotheses: carapids have (1) a mechanical barrier against the toxin constituted by a larger secretion of mucus than other fishes, (2) a bioactive barrier against the toxins constituted by a mucus effective on saponins and (3) a Δ7sterol tissue composition mimicking holothuroids that enable them to resist to saponins. First experiments showed that the mucus has no effective impact on saponin chemical structures. Mass spectrometry analyses showed that carapids, similarly to non-symbiotic fishes but contrary to their hosts, present a Δ5sterol tissue composition. However, two different procedures have shown that carapids produce six to ten times more mucus than control fishes, suggesting that a great quantity of mucus can protect carapids from their host’s saponins and acts as a mechanical barrier against toxins. Therefore, these results provide a new understanding of the carapids–holothuroids relationship.

Direct Visualization of Mucus Production by the Cold-Water Coral Lophelia pertusa with Digital Holographic Microscopy

Lophelia pertusa is the dominant reef-building organism of cold-water coral reefs, and is known to produce significant amounts of mucus, which could involve an important metabolic cost. Mucus is involved in particle removal and feeding processes, yet the triggers and dynamics of mucus production are currently still poorly described because the existing tools to study these processes are not appropriate. Using a novel microscopic technique—digital holographic microscopy (DHM)–we studied the mucus release of L. pertusa under various experimental conditions. DHM technology permits μm-scale observations and allows the visualization of transparent mucoid substances in real time without staining. Fragments of L. pertusa were first maintained in flow-through chambers without stressors and imaged with DHM, then exposed to various stressors (suspended particles, particulate food and air exposure) and re-imaged. Under non-stressed conditions no release of mucus was observed, whilst mucus strings and sheaths were produced in response to suspended particles (activated charcoal and drill cuttings sediment) i.e. in a stressed condition. Mucus strings and so-called ‘string balls’ were also observed in response to exposure to particulate food (brine shrimp Artemia salina). Upon air-exposure, mucus production was clearly visible once the fragments were returned to the flow chamber. Distinct optical properties such as optical path length difference (OPD) were measured with DHM in response to the various stimuli suggesting that different mucus types are produced by L. pertusa. Mucus produced to reject particles is similar in refractive index to the surrounding seawater, suggesting that the energy content of this mucus is low. In contrast, mucus produced in response to either food particle addition or air exposure had a higher refractive index, suggesting a higher metabolic investment in the production of these mucoid substances. This paper shows for the first time the potential of DHM technology for the detection, characterization and quantification of mucus production through OPD measurements in L. pertusa.

Ethno-medicinal and Ayurvedic Approach in the Management and Treatment of Asthma (Swash Roga)

In Ayurveda, Asthma is known as 'Shwas Roga'. Samprapti (Pathogenesis) of the Shwas Roga according to Ayurveda is the vitiated 'Pranvayu' combined with vitiated 'Kapha dosha' in the lungs causing obstruction in the 'Pranavaha srotasa' (Respiratory system). This results in gasping and laboured breathing which is known as 'Shwas Roga'. Asthma is a chronic illness involving the respiratory system in which the airway occasionally constricts, becomes inflamed, and is lined with excessive amounts of mucus, often in response to one or more triggers. These episodes precipitate by exposure to an environmental stimulant (or allergen), cold air, warm air, moist air, exercise or exertion, or emotional stress. Ayurvedic medicines are very safe and cure the problem to a great extent. Researchers of various disciplines are working on this problem to find out the solutions. Various modern means and measures have been discovered in this regard. Even then the effective drug without any side-effects has not been established yet. Ayurveda is the rich source of the therapeutic measures that can control the disease. Out of such therapeutic measures, various herbs, poly-herbal and herbo-mineral compounds were selected in different studies for the benefit of the increasing number of asthma patients and have been found to be effective. The aims of the present review are to establish the importance of Ethno-medicinal and Ayurvedic approach in the management and treatment of Asthma and explore any new interventions needed.

Pathologic Features of Colorectal Inflammatory Polyps in Miniature Dachshunds.

The histopathologic characteristics of colorectal inflammatory polyps that formed in Miniature Dachshunds were compared with those of other colorectal proliferative lesions, including adenomas and adenocarcinomas. Fifty-three colorectal polypoid lesions were histopathologically classified into inflammatory polyps (26 cases), adenoma (18 cases), and adenocarcinoma (9 cases). All 26 dogs that were diagnosed with inflammatory polyps were Miniature Dachshunds, indicating that colorectal inflammatory polyps exhibit a marked predilection for this breed. The inflammatory polyps had complex histopathologic features and were classified into 3 stages based on their epithelial composition. In early stage (stage 1), the polyps tended to exhibit a thickened mucosa containing hyperplastic goblet cells, dilated crypts filled with a large amount of mucus, and mild lymphocyte and macrophage infiltration. In later stages (stages 2 and 3), more severe neutrophil infiltration, interstitial mucus accumulation, granulation tissue, and occasional osteoid tissue were seen. Also, a few small foci of dysplastic epithelial cells were detected. The hyperplastic goblet cells, which were a major component of the epithelium of the inflammatory polyps, were positive for cytokeratin 20 (CK20), while the dysplastic epithelial cells found in inflammatory polyps (stage 3) and the tumor cells of the adenomas and adenocarcinomas were negative for CK20. These CK20-negative epithelial cells exhibited cytoplasmic and nuclear immunoreactivity for beta-catenin. In addition, the epithelial cells in the inflammatory polyps demonstrated significantly higher cyclooxygenase 2 and fibroblast growth factor 2 expression than did those of the adenomas and adenocarcinomas, suggesting that the arachidonate cascade is involved in the development of colorectal inflammatory polyps in miniature dachshunds.

Morphophysiological characterization of rhizobia isolated from wild genotypes of the common bean

This study aimed to characterize, based on the morphology and on the use of carbon sources, rhizobial isolates obtained from nodules of wild common bean genotypes cultivated on soil samples from the states of Goias, Minas Gerais and Parana. Based on morphological and carbon source use, similarity matrixes were generated to perform clustering analysis based on the Jaccard index. A total of 523 isolates were obtained with the predominance of isolates that acidify the culture medium and showed fast growth. Isolates from the soils of Araucaria and Prudentopolis produced a greater amount of mucus than those from Jussara, Nova Veneza, Uberlândia and Unai soils. The rhizobia isolates obtained from soils of Goias and Minas Gerais showed greater phenotypic diversity than those from the soils of Parana. Analysis based on the use of different carbon sources revealed that rhizobia isolates obtained from soils of Goias have higher metabolic diversity. Overall, the most part of isolates grouped with the reference strains, especially with the Rhizobium tropici and R. freirei strains used as inoculants for the common bean.

Gastroprotective effects of thymol on acute and chronic ulcers in rats: The role of prostaglandins, ATP-sensitive K+ channels, and gastric mucus secretion

Thymol, a monoterpene phenol derivative of cymene, is found in abundance in the essential oils of Thymus, Origanum, and Lippia species. The present study investigated the gastroprotective actions of thymol (10, 30, and 100 mg/kg, p.o.) in the acute (ethanol- and nonsteroidal anti-inflammatory drug-induced ulcers) and chronic (acetic acid-induced ulcers) ulcer models in rats. Some of the mechanisms underlying to the gastroprotective effect of thymol were investigated in the ethanol-induced ulcer model. Gastric secretion parameters (volume, pH, and total acidity) were also evaluated by the pylorus ligature model, and the mucus in the gastric content was determined. The anti-Helicobacter pylori activity of thymol was performed using the agar-well diffusion method. Thymol (10, 30, and 100 mg/kg) produced dose dependent reduction (P < 0.01) on the total lesion area in the ethanol-induced ulcer model. The gastroprotective response caused by thymol (30 mg/kg) was significantly attenuated (P < 0.001) by intraperitoneal treatment of rats with indomethacin (a non-selective inhibitor of cyclo-oxygenase, 10 mg/kg) and glibenclamide (ATP-sensitive K+ channel blocker, 10 mg/kg), but not by dl-Propargylglycine (PAG, a cystathionine-γ-lyase inhibitor, 25 mg/kg) and Nw-nitro-l-arginine methyl ester hydrochloride (l-NAME, a non-selective inhibitor of nitric oxide synthase, 70 mg/kg). Thymol (30 and 100 mg/kg) also reduced the ulcer index (P < 0.05) and the total lesion area (P < 0.001) in the indomethacin- and acetic-acid-induced ulcer models, respectively. In the model pylorus ligature, the treatment with thymol failed to significantly change the gastric secretion parameters. However, after treatment with thymol (30 and 100 mg/kg), there was a significant increase (P < 0.01) in mucus production. Thymol no showed anti-H. pylori activity in vitro. Collectively, the present results provide convincing evidence that thymol displays gastroprotective actions on the acute and chronic ulcer models through mechanisms that involve increased in the amount of mucus, prostaglandins, and ATP-sensitive K+ channels.

54 IMMUNOLOCALIZATION OF STEROID SULFATASE AND ESTROGEN-SPECIFIC SULFOTRANSFERASE IN BOVINE FOLLICLES

Steroid hormones are regulators in the fine-tuned mechanism of follicular development in cattle. Their concentration and property can be modulated via different processes. Sulfoconjugation via sulfotransferases (SULT) changes them from being hydrophobic to hydrophilic molecules, thereby preventing them from diffusing freely across the lipid bilayer and necessitating a transport system like the sodium-dependent organic anion transporter (SOAT; SLC10A6). In addition, sulfated steroids can no longer bind to their nuclear receptors, rendering them biologically inactive. Steroid sulfatase (STS) removes the sulfate moiety from conjugated steroids, transforming them to the free active forms. Data about the concentration of sulfated steroids in follicular fluid and the expression of the related enzymes are limited in horses and cattle. Recently, transcripts of the corresponding enzymes have been detected in cumulus cells of immature bovine cumulus-oocyte complex. Furthermore, it has been shown that small amounts of sulphated steroids are present in bovine follicular fluid. The objective of the study was to investigate the steroid metabolizing enzymes at the protein level via immunohistochemistry (IHC). Bovine ovaries collected at a slaughterhouse were categorized according to their oestrus cycle stage. Three pairs of ovaries of each cycle stage (proestrus, oestrus, postestrus, interestrus) were collected. Morphological criteria (ovaries: presence/absence of follicles and/or CL, size and number of follicles; closure of the cervix, amount of mucus) were employed to categorize them. Samples for IHC were fixed in formalin. After washing and dehydration, the samples were finally embedded in paraffin and mounted onto blocks. Indirect immunoperoxidase staining methods were applied using the streptavidin-biotin technique for signal enhancement following standard procedures. Tissue sections of 3 µm were mounted onto glass slides, and a polyclonal rabbit anti-human STS and a polyclonal rabbit anti-bovine SULT1E1 primary antibody were used to detect STS or SULT1E1, respectively. Negative controls were included using nonspecific rabbit IgG. The evaluation of the staining was descriptive in relation to a positive control (bovine placenta). Immunostaining for STS was detected in granulosa cells of antral and secondary follicles and in the endothelium of blood vessels, irrespective of the stage of the oestrus cycle. Moreover, staining for SULT1E1 was restricted to granulosa cells in antral follicles, again irrespective of the stage of the oestrus cycle. However, staining intensity for STS and SULT1E1 was only weak. These data indicate for the first time the presence of the steroid metabolising enzymes STS and SULT1E1 in bovine ovaries at the protein level. However, due to the low expression level, further studies are needed to clarify the function. We gratefully acknowledge the financial support of the German Research Foundation (DFG; FOR 1369, WR 154/3–1).

Clinical Characteristics of Dry Eye Patients With Chronic Pain Syndromes

To investigate clinical characteristics of dry eye disease (DED) patients with a chronic pain syndrome. Cross-sectional study. Four hundred twenty-five patients of a tertiary care DED patient cohort in the Netherlands were included. Chronic pain syndromes irritable bowel syndrome, chronic pelvic pain, and fibromyalgia were assessed by questionnaires. Outcome variables were the Ocular Surface Disease Index (OSDI) symptom questionnaire, tear osmolarity, Schirmer test, tear breakup time, conjunctival hyperemia, staining of the cornea and conjunctiva, and amount of mucus. Outcomes were cross-sectionally compared between DED patients with a chronic pain syndrome and those without. A total of 74 out of 425 DED patients (17%) had at least 1 chronic pain syndrome. The total symptom score was significantly higher in DED patients with a chronic pain syndrome than in those without (45.8 vs 33.8, P < .0005). Moreover, patients with a chronic pain syndrome scored higher on every single subscale of the 12-item OSDI symptom questionnaire. However, ocular signs were similar or even less severe in these patients. Similarly, in 64 DED patients from the population-based cohort TwinsUK, patients with a chronic pain syndrome (n= 24, 38%) had higher subscale and total (34.1 vs 14.4, P= .001) symptom scores. In DED patients, chronic pain syndromes are common and are associated with increased severity of DED symptoms across all domains of the OSDI, even though objective ocular surface signs are no worse. In clinical practice, more awareness of chronic pain syndromes might help in understanding the discrepancy between signs and symptoms in DED.