Effect of atractylenolide III on interstitial cells of Cajal and C-kit/SCF pathway of rats with loperamide-induced slow transit constipation

Purpose: To determine the effect of atractylenolide-III (ATL-III) on loperamide-induced slow transit constipation (STC) in a rat STC model, and to elucidate the mechanisms involved. Methods: Male Wistar rats were divided into five groups (n=6 per group): normal control group (NG), model group, and three STC rat groups treated with different doses of ATL-III, viz, 5, 10 and 15 mg/kg. The rats were treated for 15 days. Feed consumption, fecal excretion and intestinal transit rate were determined. Nitric oxide synthase (NOS), somatostatin (SS), serotonin (5-HT), and vasoactive intestinal peptide (VIP) were measured with enzyme-linked immunosorbent assay (ELISA). The protein and mRNA expressions of C-kit, SCF, PKC, and PI-3K were assayed using Western blot analysis and realtime reverse transcription polymerase chain reaction (RT-PCR), respectively. Results: The amount, weight, and moisture content of stool, and water consumption were significantly higher in ATL-III-treated groups than in the untreated (model) group (p < 0.05), whereas no difference was observed in feed intake. Intestinal transit rate was higher in the ATL-III-treated groups (p < 0.05). Decreased NOS, SS and VIP levels and increased 5-HT level were seen in the ATL-III-treated groups (p < 0.05). ATL-III treatment also induced increases in smooth muscle cells, neuronal cells, and mucous layer (p<0.05). Results from RT-PCR and Western blot revealed that ATL-III–treated groups had elevated c-kit, SCF, PKC, as well as PI-3K mRNA and protein expressions (p < 0.05). Conclusion: These results suggest that ATL-III mitigates loperamide-induced STC in rats via stimulation of NOS, SS, VIP, and 5-HT secretions. It also increases smooth muscle cells, neuronal cells, and mucous layer, and regulates the signaling pathways involving PKC, PI3K, SCF, and c-kit.

P687 Development of a panel of microbial markers to distinguish transient from pathological dysbiosis

Background Dysbiosis is a widely used but unspecific term. It has been defined as any change in the composition of resident microbial communities relative to the ones found in healthy individuals. But it is still unclear which are the appropriate communities to define it and the reference values to measure it. Different studies have described dysbiosis in various diseases such as Inflammatory Bowel Disease (IBD), diabetes, rheumatoid arthritis, and autism, among others. Besides, the microbiota can be altered by transient factors such as antibiotics, diet, stress, or infections. Therefore, different degrees of severity can be associated with the term dysbiosis from which pathological and transient dysbiosis can be differentiated. This work aimed at defining more specifically the term intestinal dysbiosis and to differentiate both transient and pathological dysbiosis. Methods Fifteen key microbial markers belonging to the principal families, classes and orders found in the human intestinal microbiota were accurately selected based on its functionality: F. prausnitzii (Fpra), E. coli (Eco), Firmicutes (Fir), Bacteroidetes (Bac), A. muciniphila (Akk), Ruminococcus sp. (Rum), Roseburia sp. (Ros), Gammaproteobacteria (Gam), Clostridia cluster I (Clo), Clostridia cluster XIV (XIV), Enterococcus sp., Lactobacillus sp. (Lac), C. albicans (Can), M. smithii (Msm), and the total bacterial load (Eub). The dysbiosis was defined using stool samples in a cohort of healthy subjects (n=24) and then validated with 9 patients diagnosed with intestinal diseases 4 IBD and 2 Irritable Bowel Syndrome (IBS)). Total DNA was extracted and the abundance of microbial markers was analysed by qPCR. Together with the establishment of the most common range in which each microbial marker was found, an index to define the pathological dysbiosis was calculated. Results Almost all healthy subjects analysed presented one or two slightly altered markers. The affected microbial markers in this “transient dysbiosis” were Fpra, Akk, and Ros, which are indicative of the mucous layer state, Firm as indicative of the lifestyle and fiber intake, Gam as characteristic of the pro-inflammatory state of the gut, and Msm as an altered intestinal habit and gas production. All patients analyzed (IBD and IBS) presented alterations mainly of the bacteria inhabiting the mucosa and an alteration of the opportunistic species related to the disrupted mucous layer and defining “pathological dysbiosis”. Conclusion This study establishes an appropriate abundance range of key microbial markers in the gut, leading to a specific definition of dysbiosis which allows to differentiate the pathological from the transient dysbiosis. These results need further validation in a larger patient cohort.

An Investigation into the Relationship between Xanthan Gum Film Coating Materials and Predicted Oro-Esophageal Gliding Performance for Solid Oral Dosage Forms

Oral drug therapy is generally provided in the form of solid oral dosage forms (SODF) that have to be swallowed and move throughout the oro-esophageal system. Previous studies have provided evidence that the oro-esophageal transit of SODF depends on their shape, size, density, and surface characteristics. To estimate the impact of SODF surface coatings during esophageal transit, an in vitro system was implemented to investigate the gliding performance across an artificial mucous layer. In this work, formulations comprised of different slippery-inducing agents combined with a common film forming agent were evaluated using the artificial mucous layer system. Xanthan gum (XG) and polyethylene glycol 1500 (PEG) were applied as film-forming agents, while carnauba wax (CW), lecithin (LE), carrageenan (CA), gellan gum (GG) and sodium alginate (SA), and their combination with sodium lauryl sulfate (SLS), were applied as slippery-inducing components. All tested formulations presented lower static friction (SF) as compared to the negative control (uncoated disc, C, F0), whereas only CW/SLS-based formulations showed similar performance to F0 regarding dynamic friction (DF). The applied multivariate analysis approach allowed a higher level of detail to the evaluation and supported a better identification of excipients and respective concentrations that are predicted to improve in vivo swallowing safety.

Cytotoxicity Studies on Naproxen and Piroxicam Nanoformulations

Caco-2 cells were used as in vitro models to assess the cell viability characteristics of the carriers Soluplus®, Gelucire 50/13 and PVP K25 and the nanoformulations of Naproxen and Piroxicam. The assessment of cell viability was done using the tetrazolium salt based MTT assay. Gelucire 50/13 and its NFs were observed to have slightly higher cytotoxicity than PVP and Soluplus® and their respective NFs. All the NFs were observed to follow the cytotoxicity trend of the polymers. Our results show that no significant decrease in cell viability was seen until 0.01% concentration of Gelucire 50/13 for 12-h exposure. The NFs as well as the polymers alone had no significant effect on the viability of Caco-2 cells below 0.01% concentrations. The intestine has a protective mucous layer, whereas the cell culture monolayers do not. The intestinal tissues also have more capacity to recover from trauma than the cultured cells. Hence the present NFs can be expected to show lesser cytotoxicity when subjected to in vivo studies.

Septicemic Infection Suspect In Jenkins' Whipray Pateobatis jenkinsii (Annandale, 1909): A Case Report

A wild-captive male Jenkins' whipray was found dead in a quarantine tank with a clinical sign before death in the form of decreased appetite for a week. The treatment history was oral administration of enrofloxacin antibiotic tablets. The therapy period lasts for ten days. The last treatment was the administration of Hepavit® (liver extract) and intramuscular injection of enrofloxacin antibiotic. One day before the death, blood was collected and then examined for the hematocrit and some parameters of chemical blood. The results of blood examination were found a decrease in blood urea nitrogen (BUN), alkaline phosphatase (ALP), and alanine aminotransferase (ALT) levels, increased glucose level, decreased total protein and albumin levels, and increased globulin level. Anatomical pathology examination was found lesions on the tail, around the eyes, and claspers. Hemorrhagic lesions were found in the mucous layer of the esophagus, stomach, and spiral colon. The blood clot was found under the tunica layer of testicular organs. The liver is damaged by showing a non-homogeneous coloration, organ thickening, congestion, and fragile consistency. Based on the results of the blood examination and was supported by the results of the anatomical pathology examination after death, it is suspected that the fish died due to the condition of septicemia infection during the previous few weeks.

Ultrosonogarphic and histological charac-teristics of the digestive tract in dogs with parvoviral enteritis

Since the clinical manifestation of enteri-tis of various etiologies has a similar picture, and parvovirus enteritis has a high mortality rate, early diagnosis of this disease is im-portant. A number of publications demon-strate changes in ultrasound parameters dur-ing inflammatory bowel diseases. The article is devoted to comparison of ultrasound and histological changes in the wall of the stom-ach and small intestine with parvovirus en-teritis in order to distinguish the most spe-cific ultrasound markers. Object of the study – 53 dogs of mixed breeds of both sexe with a confirmed diagnosis of parvovirus enteri-tis by polymerase chain reaction in real time from the age of 6 weeks to 7 months. Re-search were conducted at the Stavropol Vet-erinary Center named after Pirogov and Vet-erinary Center of the Stavropol State Agrari-an University, by the use of SIUI Apogee 1100 Omni scanner (Shantou Institute of Ultrasonic Instruments Co., Ltd., Guang-dong, China), according to the generally accepted method using a multi-frequency linear sensor with a frequency of 5-13 MHz in B-mode. The most characteristic ultrason- ic manifestations of canine parvovirus enteri-tis are hypotension and ectasia of the stom-ach and loops of the small intestine with the presence of anechogenic fluid content in their cavity; thickening of the surface layer of the mucosa in the form of a hyperechoic strip, a decrease in the ratio of the mucous layer to the entire wall thickness of the duo-denum and jejunum by more than 2 times and increased echogenicity of the mucous layer. This picture is a result of the partial necrosis and desquamation of villi, prolifera-tion of cell infiltrate in the mucous layer itself, consisting of lymphocytes, macro-phages, a small number of histiocytes and fibroblasts.

Evaluasi Pola Makan sebagai Upaya Pengurangan Kambuh pada Penderita Gastritis Usia Remaja

Gastritis is an inflamation that occur in the stomach mucous layer. It can happens because improper dietary habits that include frequency of meals, type and amount of food. The aim of this study is to evaluate dietary habits as an attempt to reduce relapse frequency in gastritis sufferer of adolescent. The study was descriptive using purposive sampling technique. Instrument used in this research was questionnaire. The research was held on June 2019 with 12 respondents of karang taruna Sinar remaja organization. This study found that more than a half of respondent have Improper dietary habits especially the type of food. They consumed irritative food that cause increasing the frequency of relapse. On the other hand, frequency of meals and amount of food don’t have direct effect on frequency relapse of gastritis

Parasites and Fungi Characteristics on Short Finned Eel Anguilla marmorata in Central Sulawesi, Indonesia

Parasitic infections are often not the direct cause of death of fish but the presence of wounds in the fish's body due to parasitic attacks is a trigger factor for secondary infection. Secondary infections can be caused by fungi, bacteria or viruses, which ultimately cause the death of fish. This study aimed to determine the characteristics of parasites and fungi that infected eel (Anguilla sp.) in the waters of Poso, Buol, Toli-toli and Donggala, Central Sulawesi as an important part of the diagnosis of fish disease in the framework of teraupetic strategies. Eel were taken as many as 30 individuals / location for observation of parasites and fungi. Parasitological examination was carried out for external and internal parasites on the mucous layer of the body, gills, intestines and stomach, while fungal isolations were carried out on muscles, skin and tissues that have abnormalities. After identification, the prevalence and intensity were carried out. The results showed that the highest prevalence of parasites were Camallanus sp (70%), Proteacephalus sp (50%) and Gyrodactylus sp (40%), and the nematode Camallanus sp the tapeworm Proteocephalus sp had the highest intensity of 57.5 and 30.8 respectively. Fungal prevalence were found highest in Saprolegnia sp (36%), and Fusarium sp (32%).