Small Intestinal Tissue Samples Research Articles

Effect of air-liquid interface on cultured human intestinal epithelial cells.

The intestinal epithelium is a dynamic barrier that allows the selective exchange of ions, hormones, proteins, and nutrients. To accomplish this, the intestinal epithelium adopts a highly columnar morphology which is partially lost in submerged culturing systems. To achieve this, small intestinal tissue samples were utilized to obtain human intestinal crypts to form enteroids. The Transwell system was subsequently employed to form a monolayer of cells that was cultured in either the submerged condition or the air-liquid Interface (ALI) condition. We found that the human intestinal monolayer under the ALI condition exhibited morphology more similar to the normal intestinal epithelium. F-actin localization and brush border formation were observed apically, and the integrity of the tight junctions was preserved in the ALI condition. Fewer apoptotic cells were observed in the ALI conditions as compared to the submerged conditions. The monolayer of cells expressed a higher level of secretory cell lineage genes in the ALI condition. The ALI condition positively contributes toward a more differentiated phenotype of epithelial cells. It serves as an amplifier that enhances the existing differentiation cue. The ALI system provides a more differentiated platform to study intestinal function compared to submerged conditions.

Application of Machine Learning in Predicting Hepatic Metastasis or Primary Site in Gastroenteropancreatic Neuroendocrine Tumors.

Gastroenteropancreatic neuroendocrine tumors (GEP-NETs) account for 80% of gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs). GEP-NETs are well-differentiated tumors, highly heterogeneous in biology and origin, and are often diagnosed at the metastatic stage. Diagnosis is commonly through clinical symptoms, histopathology, and PET-CT imaging, while molecular markers for metastasis and the primary site are unknown. Here, we report the identification of multi-gene signatures for hepatic metastasis and primary sites through analyses on RNA-SEQ datasets of pancreatic and small intestinal NETs tissue samples. Relevant gene features, identified from the normalized RNA-SEQ data using the mRMRe algorithm, were used to develop seven Machine Learning models (LDA, RF, CART, k-NN, SVM, XGBOOST, GBM). Two multi-gene random forest (RF) models classified primary and metastatic samples with 100% accuracy in training and test cohorts and >90% accuracy in an independent validation cohort. Similarly, three multi-gene RF models identified the pancreas or small intestine as the primary site with 100% accuracy in training and test cohorts, and >95% accuracy in an independent cohort. Multi-label models for concurrent prediction of hepatic metastasis and primary site returned >98.42% and >87.42% accuracies on training and test cohorts, respectively. A robust molecular signature to predict liver metastasis or the primary site for GEP-NETs is reported for the first time and could complement the clinical management of GEP-NETs.

7-Hydroxy-6-Methoxycoumarin 8-Glucoside (Fraxin) Confers Protection against Ionizing Radiation-Induced Intestinal Epithelial Cell Death Both In Vitro and In Vivo

The small intestine is highly sensitive to ionizing radiation. Radiation-induced intestinal injury (RIII) limits the therapeutic effect of tumor radiotherapy. However, there are currently few effective treatments available for the prevention or mitigation of RIII. Fraxin, structurally a derivative of a coumarin glucoside extracted from the herbal Cortex Qinpi, has the advantages of low toxicity and relatively low cost. Some studies have proven that Fraxin has anti-inflammatory and antibacterial effects. Our results show that Fraxin can not only promote the regeneration of intestinal stem cells and repair the intestinal structure caused by irradiation but also regulate the composition of gut microbiota and then play an immunomodulatory role through IL-22 signaling pathways, resulting in radioprotective effects. Human intestinal epithelial cells (HIECs) and intestinal organs were used as experimental models in vitro. The experimental group was irradiated with 6 Gy, detected by a cell counting kit, cell cloning, apoptosis, γ-H2AX and 8-OHdG IF. Male C57BL/6 mice aged 6-8 weeks were given a single dose of whole abdominal irradiation (WAI) at 13 Gy, detected by HE staining, IHC, IF and TUNEL methods, and survival and body weight changes were recorded, fecal samples and small intestinal tissue samples were collected 6 hours and 3 days after irradiation, and the gut microbiota was detected by 16S rRNA sequencing. Whole small intestinal tissue was analyzed by genomic transcript spectrum and RNA sequencing. Compared with vehicle treatment, Fraxin administration significantly improved the quality of life after WAI and maintained the body weight of mice. In addition, the crypt-villus architecture of the intestinal tracts in mice treated with Fraxin was well preserved, and the number of goblet cells, Lgr5+ intestinal stem cells (ISCs) and their daughter cells, Ki67+ proliferating cells increased significantly, reducing the structural damage caused by radiation. Meanwhile, Fraxin improved the expression of tight junction proteins such as E-cadherin and Claudin-3 after WAI, strikingly reduced the levels of FITC-dextran in serum, maintained the integrity of the intestinal barrier. In addition, we have confirmed that Fraxin can increase the cell viability of HIECs, reduce the 8OHdG fluorescence intensity induced by IR, and reduce γ H2AX lesions and apoptosis. Fraxin administration recovered the disorder of the structure of gut microbiota after irradiation and promoted the expression of probiotics, and KEGG pathway analysis suggested that it was related to immunity. Transcriptional sequencing showed that the IL-22 signaling pathway was significantly activated in the Fraxin treatment group, suggesting that Fraxin may have a protective effect on intestinal radiation through IL-22. In conclusion, Fraxin treatment attenuate RIII by modulating the composition of the gut microbiota and activate the IL-22 signaling pathway to reduce RIII.

Surveillance for SARS-CoV-2 in Norway Rats (Rattus norvegicus) from Southern Ontario

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from wildlife has raised concerns about spillover from humans to animals, the establishment of novel wildlife reservoirs, and the potential for future outbreaks caused by variants of wildlife origin. Norway rats (Rattus norvegicus) are abundant in urban areas and live in close proximity to humans, providing the opportunity for spillover of SARS-CoV-2. Evidence of SARS-CoV-2 infection and exposure has been reported in Norway rats. We investigated SARS-CoV-2 infection and exposure in Norway rats from Southern Ontario, Canada. From October 2019 to June 2021, 224 rats were submitted by collaborating pest control companies. The majority of samples were collected in Windsor (79.9%; n = 179), Hamilton (13.8%; n = 31), and the Greater Toronto Area (5.8%; n = 13). Overall, 50.0% (n = 112) were female and most rats were sexually mature (55.8%; n = 125). Notably, 202 samples were collected prior to the emergence of variants of concern (VOC) and 22 were collected while the Alpha variant (B.1.1.7) was the predominant circulating VOC in humans. Nasal turbinate (n = 164) and small intestinal (n = 213) tissue samples were analyzed for SARS-CoV-2 RNA by RT-PCR. Thoracic cavity fluid samples (n = 213) were tested for neutralizing antibodies using a surrogate virus neutralization test (sVNT) (GenScript cPass); confirmatory plaque reduction neutralization test (PRNT) was conducted on presumptive positive samples. We did not detect SARS-CoV-2 RNA in any samples tested. Two out of eleven samples positive on sVNT had neutralizing antibodies confirmed positive by PRNT (1 : 40 and 1 : 320 PRNT70); both were collected prior to the emergence of VOC. It is imperative that efforts to control and monitor SARS-CoV-2 include surveillance of rats and other relevant wildlife species as novel variants continue to emerge.

Effects of clarithromycin on the pharmacokinetics of tacrolimus and expression of CYP3A4 and P-glycoprotein in rats.

The objective of this study was to investigate the effect of clarithromycin on the pharmacokinetics of tacrolimus in rats and better understand its mechanism. In the control group (n = 6), rats received a single oral dose of 1mg tacrolimus on day 6. In the experimental group (n = 6), rats received 0.25 g of clarithromycin daily for five consecutive days and then a single oral dose of 1mg tacrolimus on day 6. Orbital venous blood (250 μL) was collected at 0, 0.25, 0.50, 0.75, 1, 2, 4, 8, 12, and 24 h before and after tacrolimus administration. Blood drug concentrations were detected via mass spectrometry. Small intestine and liver tissue samples were collected after rats were euthanized via dislocation, and CYP3A4 and P-glycoprotein (P-gp) protein expression was determined using western blotting. Clarithromycin increased the blood tacrolimus concentration and affected its pharmacokinetic properties in rats. Compared with those in the control group, the AUC0-24 , AUC0-∞ , AUMC(0-t) , and AUMC(0-∞) of tacrolimus in the experimental group were significantly increased, whereas the CLz/F was significantly lower (P < 0.01). Simultaneously, clarithromycin significantly inhibited CYP3A4 and P-gp expression in the liver and intestine. Protein expression of CYP3A4 and P-gp in the liver and the intestinal tract was significantly downregulated in the intervention group compared with that in the control group. Clarithromycin significantly inhibited the protein expression of CYP3A4 and P-gp in the liver and intestine, thereby increasing the mean blood concentration and significantly increasing the AUC of tacrolimus.

Histopathological and ultra-structural investigation of the damaging effects of hypoinsulinemia, hyper glycaemia and oxidative stress caused by parenteral nutrition combined with fasting on the small intestine of rabbits

Background and study aimsParenteral nutrition (PN) is a life-saving practice when the use of the gastrointestinal tract is not appropriate. Despite its great benefits, however, PN may cause several complications. In this study, we conducted histopathological and ultra-structural examinations of the effect of PN combined with starvation on the small intestines of rabbits. Materials and methodsRabbits were divided into four groups. A fasting + PN group was left completely unfed and received all its daily required energy by PN through an intravenous central catheter. An oral feeding + PN group received half the necessary daily calories by oral feeding and the other half through PN. A semi-starvation group received only half the necessary daily calories by oral feeding and no PN. The fourth group, serving as a control, was supplied with its entire daily energy requirements through oral feeding. After 10 days, the rabbits were euthanized. Blood and small intestine tissue samples were collected from all groups. Blood samples were biochemically analysed, and tissue samples were examined by light and transmission electron microscopy. ResultsThe fasting + PN group exhibited lower insulin levels, higher glucose levels, and increased systemic oxidative stress than the other groups. Ultra-structural and histopathological examinations revealed a significant increase in apoptotic activity in this group’s small intestines and a significant decrease in villus length and crypt depth. Severe damage to the intracellular organelles and nuclei of enterocytes was also observed. ConclusionPN combined with starvation appears to cause apoptosis in the small intestine due to oxidative stress and hyperglycaemia with hypoinsulinemia, with destructive effects on small intestine tissue. Adding enteral nutrition to PN may reduce these destructive effects.

Peroxiredoxins and Hypoxia-Inducible Factor-1α in Duodenal Tissue: Emerging Factors in the Pathophysiology of Pediatric Celiac Disease Patients.

Celiac disease (CD) is an autoimmune enteropathy. Peroxiredoxins (PRDXs) are powerful antioxidant enzymes having an important role in significant cellular pathways including cell survival, apoptosis, and inflammation. This study aimed at investigating the expression levels of all PRDX isoforms (1-6) and their possible relationships with a transcription factor, HIF-1α, in the small intestinal tissue samples of pediatric CD patients. The study groups consisted of first-diagnosed CD patients (n = 7) and non-CD patients with functional gastrointestinal tract disorders as the controls (n = 7). The PRDXs and HIF-1α expression levels were determined by using real-time PCR and Western blotting in duodenal biopsy samples. It was observed that the mRNA and protein expression levels of PRDX 5 were significantly higher in the CD patients, whereas the PRDX 1, -2, and -4 expressions were decreased in each case compared to the control group. No significant differences were detected in the PRDX 3 and PRDX 6 expressions. The expression of HIF-1α was also significantly elevated in CD patients. These findings indicate, for the first time, that PRDXs, particularly PRDX 5, may play a significant role in the pathogenesis of CD. Furthermore, our results suggest that HIF-1α may upregulate PRDX-5 transcription in the duodenal tissue of CD.

Introduction of a pharmacological neurovascular uncoupling model in rats based on results of mice.

Our aim was to establish a pharmacologically induced neurovascular uncoupling (NVU) method in rats as a model of human cognitive decline. Pharmacologically induced NVU with subsequent neurological and cognitive defects was described in mice, but not in rats so far. We used 32 male Hannover Wistar rats. NVU was induced by intraperitoneal administration of a pharmacological "cocktail" consisting of N-(methylsulfonyl)-2-(2-propynyloxy)-benzenehexanamide (MSPPOH, a specific inhibitor of epoxyeicosatrienoic acid-producing epoxidases, 5 mg kg-1), L-NG-nitroarginine methyl ester (L-NAME, a nitric oxide synthase inhibitor, 10 mg kg-1) and indomethacin (anonselective inhibitor of cyclooxygenases, 1 mg kg-1) and injected twice daily for 8 consecutive days. Cognitive performance was tested in the Morris water-maze and fear-conditioning assays. We also monitored blood pressure. In a terminal operation a laser Doppler probe was used to detect changes in blood-flow (CBF) in the barrel cortex while the contralateral whisker pad was stimulated. Brain and small intestine tissue samples were collected post mortem and examined for prostaglandin E2 (PGE2) level. Animals treated with the "cocktail" showed no impairment in their performance in any of the cognitive tasks. They had higher blood pressure and showed cca. 50% decrease in CBF. Intestinal bleeding and ulcerswere found in some animals with significantly decreased levels of PGE2 in the brain and small intestine. Although we could evoke NVU by the applied mixture of pharmacons, it also induced adverse side effects such as hypertension and intestinal malformations while the treatment did not cause cognitive impairment. Thus, further refinements are still required for the development of an applicable model.

Effect of the extract made from Rhizoma Atractylodis Macrocephalae (RAM) on the immune responses of mice to a commercial foot-and-mouth disease vaccine.

ObjectivesFoot‐and‐mouth disease (FMD) is an economically important animal disease because of the speed of its transmission. Routine vaccination may not be effective; RAM can be considered as a potential facilitator for this. Present study was designed to evaluate the effects of feeding different treatment of the RAM in different days on the immune responses in mice immunised with FMDV type O vaccine.Material and methodsIn experiment 1, 50 ICR mice were randomly divided into five groups with 10 animals in each group, and the basic diet containing 1% Crush of RAM for 1‐week ad libitum feeding period, 1% Crush of RAM for 6‐week ad libitum feeding period, 1% Decoction of RAM for a 1‐week ad libitum feeding period, 1% Decoction of RAM for a 6‐week ad libitum feeding period, respectively. Blood samples were collected 2 weeks after boosting for measurement of FMDV‐specific IgG level and the IgG subclasses, lymphocyte proliferation as well as production IL‐5 and IFN‐γ. In experiment 2, four groups mice were fed basic diet and basic diet containing 5% Decoction of RAM for 2‐, 4‐ and 6‐day ad libitum feeding periods, respectively. Then we collected blood samples for detecting IgG and IgG subclasses, splenocytes for lymphocyte proliferation as well as production IL‐5 and IFN‐γ, and tissue samples of small intestine for sIgA.ResultsThe results indicated that 1% Decoction of RAM for a 1‐week ad libitum feeding period group and 5% Decoction of RAM for 2‐, 4‐ and 6‐day ad libitum feeding period group enhance the FMDV‐specific immune responses significantly.ConclusionsTaken together, the results demonstrate that doses and feeding time of RAM are important to affect the immune responses.

[Comparison of distribution of verbascoside in normoxic and hypoxic rats].

This study established a high performance liquid chromatography(HPLC) method for the simultaneous determination of verbascoside(VB) and its main metabolite caffeic acid(CA) in rat tissue samples. A low-pressure low-oxygen animal experimental chamber was used to simulate the plateau environment for establishing the hypoxic rat model. After intragastric administration of 300 mg·kg~(-1) VB, the normoxic and hypoxic rats were sacrificed for the collection of heart, liver, spleen, lung, kidney, brain, muscle, large intestine, small intestine, and stomach tissue samples at the time points of 30, 60, and 90 min. VB and CA concentrations in each tissue sample were measured by HPLC, and the distribution of VB and CA in normoxic and hypoxic rats was compared. The results showed that after intragastric administration, VB can be rapidly absorbed and distributed into various tissues including brain in both normoxic and hypoxic rats, indicating that VB can pass through the blood-brain barrier. In the gastrointestinal tract, VB was mainly distributed in small intestine, which suggested that the main absorption site of VB was small intestine. A large amount of VB was detected in muscle and lung, and only a small amount in other tissues. CA was detected in other tissues except brain, heart, and muscle. Small intestine had the most abundant CA, followed by stomach, large intestine, and kidney, and only a small amount of CA was detected in the liver, spleen, and lung(&lt;50 ng·mL~(-1)). The results indicated that VB may be mainly absorbed and metabolized in the gastrointestinal tract to produce CA and was possibly excreted through kidney. Compared with normoxic rats, hypoxic rats had reduced and slow distribution of VB and increased ratio of VB concentration in tissue to plasma, which implied that the relative proportion of VB from systemic circulation to tissues was increased in hypoxic rats. This study provides a basis for the application of VB in anti-hypoxia therapy and for the formulation of anti-hypoxia dosing regimens.

Lymphocyte subsets in the small intestine of piglets fed with probiotic and zinc: a qualitative and quantitative micro-anatomical study.

Piglet mortality is a real concern to the pig farmers. The major cause is due to the late maturation of the immune system and dietary changes in postweaned piglets. The potential role of probiotic and zinc in the stimulation of the immune system is well established. Hence, the present study was undertaken to evaluate alterations of T and B cells in the small intestine after dietary inclusion of probiotic and zinc in pre and post-weaned piglets. A total of 18 healthy Large White Yorkshire (LWY) piglets, irrespective of sex obtained from 3 litters at the age-group of 20, 30 and 60 days. They were divided into a control group fed with basal diet and a treatment group fed with probiotic and zinc supplement along with the basal diet, consisting of three animals in each group. The piglets were weaned at 28 days of age. After sacrificing the animals at day 20, 30 and 60 from both the groups, the abdominal cavity was opened and small intestinal tissue samples were collected, processed and stained by indirect immunofluorescence technique. The slides were evaluated under the fluorescent light microscope. The data were statistically analysed. The different T and B cell subsets were recorded in the lining epithelium, core of villus, crypt area of lamina propria and Peyer's patch area. The number of CD4+, CD8+, IgA+ and IgM+ cells was higher in the treated piglets than the control group of animals, irrespective of segments of intestine and age-group. It can be concluded that the dietary supplementation of probiotic and zinc was found to be good additives as they can stimulate the immune response in piglets, especially during the critical early post-weaning period.

Glucagon-Like Peptide-2 Modulates Enteric Paneth Cells Immune Response and Alleviates Gut Inflammation During Intravenous Fluid Infusion in Mice With a Central Catheter.

Background: Glucagon-like peptide-2 (GLP-2) has protective effects on gastrointestinal functions. Our previous study found that GLP-2 could significantly reduce intestinal permeability and bacterial translocation in total parenteral nutrition (TPN) animal model. However, the effects of GLP-2 on the impairment of the intestinal Paneth cells immune function and gut inflammation during intravenous fluid infusion mainly consisted of nutritional materials is currently scattered.Objective: The current study was aimed to investigate the efficacy of the GLP-2 in alleviating gut inflammation and modulating enteric Paneth cells immune response in parenterally fed mice and its underlying mechanisms.Methods: Thirty-six male ICR mice underwent venous catheterization were divided into 3 groups: Chow, TPN, and TPN+GLP-2 groups. GLP-2 was administered intravenously at 60 μg/day for 5 days. The small intestine tissue and serum samples were collected on the 7th day.Results: Compared with the TPN group, the expression of tight junction proteins occludin and claudin-1 were significantly increased in the TPN+GLP-2 group. In addition, the expression of lysozyme, sPLA2, insulin-like growth factor-1, and epithelial protection and repair genes were improved in the TPN+GLP-2 group. The levels of IL-6 and TNF-α proteins and mRNAs in the ileum tissues were remarkably reduced in the TPN+GLP-2 group, while IL-10 protein and mRNA level were elevated in the TPN+GLP-2 group (all p < 0.05). Moreover, the TPN+GLP-2 group has higher levels of serum endotoxin, D-lactic acid, and MPO than those of the TPN group.Conclusions: GLP-2 alleviated gut inflammation and improved enteric Paneth cells immune responses through intravenous fluid infusion, possibly by improving the functioning of epithelial protection and repair, and reducing mucosal inflammatory responses.

Protective Effects of an Oxovanadium(IV) Complex with N2O2 Chelating Thiosemicarbazone on Small Intestine Injury of STZ-Diabetic Rats.

Vanadium compounds are being investigated as potential therapeutic agents in the treatment of many health problems, primarily diabetes. We aimed to provide the effect of N(1)-4-hydroxysalicylidene-N(4)-salicylidene-S-methyl-isothiosemicarbazidato-oxovanadium(IV) (VOL) on small intestinal injury in experimental male diabetic rats. Four groups were created of 3.0-3.5-month-old rats. The rats were made diabetic by a single dose of streptozotocin (STZ) at 65 mg/kg and grouped as follows: control animals, VOL-given control animals, STZ-induced diabetic animals and STZ-induced diabetic animals given VOL. A daily dose of 0.2 mM/kg vanadium complex was administered orally for 12 days after the inducement of diabetes. On the 12th day, small intestine tissue samples were taken. According to the data obtained from the biochemical analysis, reduced glutathione (GSH) level, catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST), superoxide dismutase (SOD), Na+/K+-ATPase and paraoxanase (PON) activities were increased, whereas sialic acid (SA), xanthine oxidase (XO) and disaccharidases (maltase and saccharidase) activities were decreased in the small intestine tissue of VOL-treated diabetic rats. Microscopic examinations revealed a remarkable decrease in the mucosal necrotic areas, discontinuity in the brush border, deterioration of the villi integrity and oedema inside the villi, but with a mild decrease in the inflammatory cells, deterioration and loss of integrity of the gland in the small intestine of VOL-treated diabetic rats. Moreover, VOL treatment markedly decreased the proliferation of villus cells and especially inflammatory cells in the small intestine of diabetic rats. According to the obtained data, the administration of VOL is a potentially convenient strategy to reducing small intestine injury in diabetic rats.

Protective effect of astaxanthin against cisplatin-induced gastrointestinal toxicity in rats

The aim of the present study was to demonstrate astaxanthin’s attenuating effects against cisplatin (CIS)-induced gastrointestinal toxicity in a rat model. Thirty-two Wistar rats (weight 200–250 g) were divided into four groups (8 rats/group): control group, CIS group, astaxanthin 25 (ASTA25) group, and astaxanthin 75 (ASTA75) group. Tissue samples of small intestine extracted from rats were histopathologically and immunohistochemically analyzed. Histopathological findings were graded with the Duodenal Histopathological Damage Score. Evaluation of enterocytes showing caspase‑3 positivity was performed using the Immunohistochemistry Positivity Score. Intestinal glutathione (GSH) shows the endogenous antioxidant level. Lipid peroxidation in the intestinal tissue was evaluated by measuring the level of malondialdehyde (MDA), which is the end product of lipid peroxidation. In the ASTA25 group, we observed a significant decrease in the number of necrotic enterocytes, mucosal ulceration, and inflammation. In the ASTA75 group, enterocytes with normal microvilli were noticeable. According to the immunohistochemical results, in the ASTA25 and ASTA75 groups, caspase‑3 positivity scores in enterocytes were lower than the CIS group. Astaxanthin administration significantly inhibited MDA increase in intestinal tissue (p < 0.005). Cisplatin administration caused a significant decrease in GSH levels. Astaxanthin showed a non-significant rise of GSH compared to the CIS group. Based on the data in this study, astaxanthin might play a protective role against cisplatin-induced gastrointestinal toxicity in rats.

Preparing the peritoneal surface for intraperitoneal chemotherapy using high-intensity ultrasound (HIUS): Investigation of technical feasibility, safety and possible limitations.

e16045 Background: The penetration of chemotherapeutic drugs into metastatic peritoneal nodules remains at levels well below 1 mm, thus significantly limiting the antitumor effect of intraperitoneal chemotherapy (IPC). Recently, high-Intensity ultrasound (HIUS) has been discovered as a potential tool to improve peritoneal diffusion rates. Despite promising preliminary data, basic aspects regarding its technical feasibility, safety and possible limitations remain unclear. This study aims to enhance our current understanding of HIUS and test its applicability using an ex-vivo swine model. Methods: Three postmortem swine were subject to laparotomy with consecutive lavage with 0.9%NaCl saline and HIUS application. For this purpose, a large HIUS radiating pen was introduced into the abdominal cavity and HIUS was applied on two of the four abdominal quadrants for 300 seconds each at an output power of 70 W, 50 % amplitude and 20 kHz frequency. Following the procedure, small intestinal tissue samples were removed for further analyses. Results: In all specimen, peritoneal and subperitoneal layers showed structural changes only visible on a microscopic level. The peritoneal layer was transformed into a mash-like structure while the subperitoneal layer (depth of 142 +/- 28 µm) exhibited microcavities and vascular detachment from surrounding tissues. No bowel rupture or vascular perforations were observed. Conclusions: Our data indicate that HIUS is a technically feasible add-on procedure for IPC with measurable microscopic changes on the peritoneal surface and no apparent bowel toxicity. Pretreatment of the abdominal cavity with HIUS could significantly improve IPC efficacy. Further studies are required to optimize and evaluate this novel approach.

Intraperitoneal chemotherapy of the peritoneal surface using high-intensity ultrasound (HIUS): investigation of technical feasibility, safety and possible limitations.

Introduction: The penetration of chemotherapeutic drugs into peritoneal nodules remains at levels well below 1 mm, thus significantly limiting the antitumor effect of intraperitoneal chemotherapy (IPC). Recently, high-Intensity ultrasound (HIUS) has been discovered as a potential tool to significantly improve peritoneal diffusion rates. Despite promising preliminary data, basic aspects regarding its technical feasibility, safety and possible limitations remain unclear. This study aims to enhance our current understanding of HIUS and test its applicability using an ex-vivo swine model.Methods: Three postmortem swine were subject to laparotomy and consecutive lavage with 0.9%NaCl saline and HIUS application. For this purpose, a large HIUS radiating pen was introduced into the abdominal cavity and HIUS was applied on two of the four abdominal quadrants for 300 seconds each at an output power of 70 W, 50 % amplitude and 20 kHz frequency. Following the procedure, small intestinal tissue samples were retrieved for further analyses.Results: Peritoneal and subperitoneal layers showed structural changes only visible on a microscopic level. The peritoneal layer was transformed into a mesh-like structure while the subperitoneal layer (depth of 142 +/- 28 µm) exhibited microcavities and vascular detachment from surrounding tissues. No bowel rupture or vascular perforations were observed.Conclusions: Our data indicate that HIUS is a technically feasible and safe add-on procedure for intraperitoneal chemotherapy (IPC) with measurable microscopic changes on the peritoneal surface. Pretreatment of the abdominal cavity with HIUS could significantly improve IPC efficacy. Further studies are required to optimize and evaluate this novel approach.

Effects of Dietary Fiber Level in Weaning Pig Diets on Growth Performance, Nutrient Digestibility and Intestinal Morphology

Insoluble dietary fiber is benefit on gut volume and morphology of the pigs. Rice hull is the by-product from rice processing that rich of dietary fiber content. The objectives of our study were to evaluate the effects of dietary fiber level from grounded rice hull on growth performance, nutrient digestibility, intestinal morphology and bacterial cell count in hindgut of weaning pigs. Thirty weaning pigs (21±3 days of age) were fed randomly with three experimental diets consisting of three levels of total Dietary Fiber (DF); 130, 140 and 150 g/kg with equal ratios of insoluble per soluble dietary fiber. On the 29th day, four pigs per treatment were euthanized for the collection of small intestinal tissue samples for morphological determination and digesta samples were collected for microbial counts. For digestibility determination, 12 weaned pigs were used to determine nutrient digestibility. The Feed Conversion Ratio (FCR) pigs at Week 1 was improved, however, there was overall no significant difference for growth performance and diarrhea incidence among the groups. The nutrient digestibility of energy and protein did not differ among treatments, whereas the digestibility of crude fat and fiber tended to increase (P = 0.082 and 0.074). Increasing of dietary fiber level tended to increase villus height and enhance villus height per crypt depth ratio (VH:CD) of ileum (VH:CD ratio were 1.94, 2.10 and 2.35 in pigs fed 130, 140 and 150 g/kg dietary fiber diets respectively; P = 0.062). The bacterial count in the gut content was not affected by dietary fiber level. In conclusion, the dietary fiber level between 130 to150 g/kg was not altering overall growth performance nutrient digestibility, intestinal morphology intestinal bacteria population. However, the dietary fiber 140-150 g/ kg in diet could improve FCR in the first week after weaning and tended to improve the ileal morphology.

Typhlocolitis by Edwardsiella tarda in a Cow

Background: Edwardsiella tarda has been report as etiology of gastroenteritis in both human and veterinary medicine, usually associated with exposure to aquatic environments in immunocompromised individuals. The present report describes a case of typhlocolitis in a cow in the west region of Santa Catarina state, Brazil. Case: After clinically evaluation of animal and proceeding euthanasia and realized the necropsy. Tissue samples were collected, and routinely processed for histological examination. Histopathological lesions were described as mild, moderate and severe. Tissue and swabs samples of small and large intestine were sent to standard microbiological culture processing. At necropsy, cattle presented severe dehydration and emaciation. Eye and vaginal mucosa were severely congested. The opening of the abdominal cavity revealed a great amount of greenish fluid and a large amount of fecal material, associated with diffuse severe peritonitis, evidenced by hyperemia and severe deposition of fibrin in the visceral and parietal peritoneal surface. In the serosa of the cecum, there were two points of rupture, observed in the proximal colon. The mucosa of cecum and colon were severely edematous, hyperemic, and presented diffusely distributed pinpoint round hemorrhages, as well as fibrinonecrotic material adhered to the surface. Histologically, in the mucosa of cecum and colon, moderate to severe diffuse inflammatory infiltrate of neutrophils, lymphocytes and plasma cells associated with multifocal severe necrosis were observed. Moderate diffuse fibrin deposition was evidenced in the submucosa and muscular, as well as multifocal moderate necrosis in the muscular layer. In the serous, severe diffuse inflammatory infiltrate of neutrophils associated with fibrin deposition and innumerous coccoid bacterial colonies were observed. The samples subjected to bacterial isolation showed growth of Edwardsiella tarda. All samples were negative for Salmonella spp. and Yersinia spp.Discussion: The final diagnosis was established through the association of clinical history, clinical signs, gross and histopathological lesions, as well as, bacterial isolation of the etiological agent, Edwardsiella tarda. In this case, it is conjectured that the reservoirs which cattle had access represented the source of infection. The fact that the animal was in the immediate postpartum period may have predisposed to the development of clinical disease due to immunosuppression. In domestic animals, Edwardsiella tarda has been reported in swine, and as a cause of septicemia in calves. Clinically, intestinal manifestations observed in edwardsiellosis in cattle are indistinguishable from several other conditions that cause diarrhea, such as infectious, nutritional or parasitic diseases. The main differential diagnoses are salmonellosis and yersiniosis due to the similarities regarding to gross and histopathological lesions in these cases compared to cases of edwardsiellosis. Salmonellosis is characterized by grey to yellowish, fetid diarrhea in which blood and mucus are oftentimes observed. At necropsy, catarrhal, hemorrhagic or fibrinous enteritis may be evidenced. The lesions initially are seen in the ileum. However, in the chronic stages of infection, foci of necrosis and ulceration may be noted mainly in the cecum and colon. Histologically, a fibrin layer associated with necrosis and mucosal ulceration can be observed in the small intestine and initial portion of large intestine. Inflammatory infiltrate composed predominantly by neutrophils, as well as fibrin thrombi in capillaries and venules are also observed. Lesions observed are similar that described in ulcerative colitis by E. tarda in human patients. In conclusion, Edwardsiella tarda can lead to a fatal typhlocolitis in cattle, being an important differential diagnosis in cases of acute diarrhea.Arya A.V., Rostom A., Dong W.F. &amp; Flynn A.N. 2011. Crohn’s Disease Exacerbation Induced by Edwardsiella tarda Gastroenteritis. Gastroenterology. 5(3): 623-627.Engel J.J. &amp; Martin T.L. 2006. Edwardsiella tarda as a cause of postdysenteric ulcerative colitis. International Journal Colarectal Disease. 21(2): 184-185.Ewing W.H., McWhorter A.C., Escobar M.R. &amp; Lubin A.H. 1965. Edwardsiella, a new genus of enterobacteriaceae based on a new species, E. tarda. 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Comparison of Oxidative Effects of Two Different Administration Form of Oxybutynin in the Potential Target Tissues.

Oxybutynin is an important anticholinergic agent that prevents uncontrolled contractions in the treatment of overactive bladder (OAB). However, drugs containing oxybutynin have significant side effects such as dry eyes, dry mouth, increased heart rate, constipation, blurred vision, and confusion. In recent years, new delivery methods for this agent are being searched. One of them is vaginal delivery. In this study, we aimed to compare the effects of oxybutynin on oxidative parameters in the potential target tissues of the oral and vaginal delivery. Female New Zealand white rabbits (n=12) were divided into two groups: oral delivery and vaginal delivery. The animals were sacrificed 48 h after administration and nitric oxide (NOx), thiobarbituric acid-reactive substances (TBARs), and glutathione (GSH) levels were determined spectrophotometrically in the aorta, salivary gland, and small intestine tissue samples. Vaginal delivery significantly decreased NOx levels in all tissue samples as compared to oral delivery (p < 0.05). Moreover, it reduced TBARs levels in salivary gland and aorta tissue samples (p < 0.05). In the light on these findings, it can be said that vaginal delivery may decrease the oxidant-induced side effects of oxybutynin as compared to oral delivery.

Effects of sugar beet pulp and roasted canola seed, on performance, rumen and small intestine digesta volatile fatty acid concentrations, and small intestine morphology of Arabian lambs

The effects of partial replacement of barley (B) with sugar beet pulp (SBP) with and without roasted canola seed (RCS) on performance, rumen fluids and small intestine digesta volatile fatty acids (VFA) concentrations, digesta viscosity, and small intestine morphology were investigated. Twenty-four Arabian male lambs (118 ± 10 d, 23.7 ± 2.5 kg BW) were fed with diets containing 1) 68% barley (B-RCS); 2) 68% B plus 7% RCS (B + RCS); 3) 36% SBP (SBP-RCS); 4) 36% SBP plus 7% RCS (SBP + RCS). Feeding (fattening) was undertaken for 99 days including 14 day adaptation period. Rumen fluid was collected on d 27, 54, and 81 of fattening period. At slaughter, small intestine tissue samples and digesta were collected. Results showed that lamb performance was not affected (P > 0.05) by carbohydrate source, however it was improved by inclusion of RCS. Lambs fed with SBP had higher (P < 0.05) total VFA and molar proportions of acetate, isobutyrate, and acetate to propionate ratio compared to those lambs fed without SBP diet. Inclusion of RCS increased (P < 0.05) propionate and decreased acetate and isobutyrate in the rumen fluid but there was no interaction of SBP × RCS treatments observed for any of the time points measured. In duodenum and jejunum digesta, lambs fed SBP had higher (P < 0.05) acetate, total VFA molar concentrations, and acetate to propionate ratio than lambs fed without SBP diet. Inclusion of RCS increased (P < 0.05) acetate, total VFA concentration in duodenum and jejunum. Both pH and viscosity of the small intestine digesta in duodenum and jejunum were higher (P < 0.05) in lambs fed SBP than those fed without SBP diet. Partial replacement of barley with SBP increased crypt depth throughout small intestine and also increased thickness of tunica muscularis and villus height in jejunum and ilium. Feeding lambs with diets containing SBP + RCS increased (P < 0.05) number of Brunner's glands, number of villi, and epithelium thickness. In conclusion, partial replacement of barley by SBP along with RCS altered molar proportion of VFA in the rumen, intestine and improved small intestinal morphology of fattening Arabian lambs.