Sections Of Small Intestine Research Articles

Successful Retrocecal Hernia Diagnosis and Treatment: A Case Report

Introduction A retrocecal hernia is a rare type of internal hernia that occasionally causes small bowel obstruction and strangulation ileus. We report a case of retrocecal hernia that was preoperatively diagnosed using multidetector raw computed tomography and successfully treated by emergency surgery. Case Presentation A 97-year-old woman presented at another hospital with abdominal distension and nausea. She was diagnosed with ileus and was conservatively treated with long intestinal tube placement. However, the symptoms had not improved after 7 days in the hospital. The patient was then referred to our hospital, where a pericecal hernia was diagnosed using multidetector raw computed tomography. We performed emergency surgery and intraoperatively confirmed the presence of a retrocecal hernia. A 30-cm nonviable section of small intestine was resected, and the hernia orifice was closed routinely. Postoperative recovery was uneventful. Conclusions Patients with pericecal hernia usually require surgical treatment. Multidetector raw computed tomography is useful for precise diagnosis of this type of hernia. Precise diagnosis is necessary to select prompt operative intervention for internal hernia.

Intestinal miRNAs regulated in response to dietary lipids

The role of miRNAs in intestinal lipid metabolism is poorly described. The small intestine is constantly exposed to high amounts of dietary lipids, and it is under conditions of stress that the functions of miRNAs become especially pronounced. Approaches consisting in either a chronic exposure to cholesterol and triglyceride rich diets (for several days or weeks) or an acute lipid challenge were employed in the search for intestinal miRNAs with a potential role in lipid metabolism regulation. According to our results, changes in miRNA expression in response to fat ingestion are dependent on factors such as time upon exposure, gender and small intestine section. Classic and recent intestinal in vitro models (i.e. differentiated Caco-2 cells and murine organoids) partially mirror miRNA modulation in response to lipid challenges in vivo. Moreover, intestinal miRNAs might play a role in triglyceride absorption and produce changes in lipid accumulation in intestinal tissues as seen in a generated intestinal Dicer1-deletion murine model. Overall, despite some variability between the different experimental cohorts and in vitro models, results show that some miRNAs analysed here are modulated in response to dietary lipids, hence likely to participate in the regulation of lipid metabolism, and call for further research.

EFFECT OF SOME HERBAL FEED ADDITIVES ON THE PERFORMANCE OF BROILER CHICKENS

The present experiment was conducted in order to investigate the effect of Moringa Oleifera Leaves powder on the performance, some blood serum components, some histomorphological measurements and economic efficiency of broiler chickens. A total of 225 Cobb unsexed one - day old broiler chicks, were distributed at random into 5 treatments each in 3 replicates, 15 chicks each. Moringa Oleifera leaves powder (MOL) was added at the levels of 0, 0.25, 0.50 and 0.75% to low crud protein diet (LCP, 20 and 17%, negative control) compared to the normal crud protein (23 and 20%, positive control), respectively at the starter (1- 21 d) and at the finisher (22- 35d) period. Results indicated that, chicks fed the low crude protein diet supplemented with Moringa Oliefera leaves powder (MOL) had significantly (P ≤ 0.05) higher body weight, body weight gain, daily feed intake and performance index. Feed conversion ratio, European efficiency index and relative economic efficiency were also improved with the supplementation. Chicks fed low crude protein diet supplemented with 0.25 or 0.50% MOL have the highest dressing weight (1753 and 1738g) with no significant differences between them compared and increased serum total protein and glucose levels, while significantly decreased serum total cholesterol and aspartic transaminase (AST) enzyme compared to the other treatments with increasing the levels of MOL up to 0.50%. Morphological sections of small intestine revealed that villi height, width and height: width ratios were increased by the supplementation of different levels of MOLM (0.25 and 0.50%) but not significant except in duodenum villi width and in the ration between height and width. In general, the obtained results indicated that supplementation of 0.25% Moringa Oliefera Leaves, MOL can be used in broiler diets with low protein level to get the heaviest weight, best feed conversion, performance index, due to an increase in small intestinal absorption and activity. This, in turn, resulted in an increase in European efficiency index and economic efficiency under the experimental conditions.

PERFORMANCE AND HISTOMORPHOLOGICAL PARAMETERS OF BROILER CHICKS FED LOW CRUDE PROTEIN DIET SUPPLEMENTED WITH MORINGA OLEIFERA LEAVES POWDER

The present experiment was conducted in order to investigate the effect of Moringa Oleifera Leaves powder on the performance, some blood serum components, some histomorphological measurements and economic efficiency of broiler chickens. A total of 225 Cobb unsexed one - day old broiler chicks, were distributed at random into 5 treatments each in 3 replicates, 15 chicks each. Moringa Oleifera leaves powder (MOL) was added at the levels of 0, 0.25, 0.50 and 0.75% to low crud protein diet (LCP, 20 and 17%, negative control) compared to the normal crud protein (23 and 20%, positive control), respectively at the starter (1- 21 d) and at the finisher (22- 35d) period. Results indicated that, chicks fed the low crude protein diet supplemented with Moringa Oliefera leaves powder (MOL) had significantly (P ≤ 0.05) higher body weight, body weight gain, daily feed intake and performance index. Feed conversion ratio, European efficiency index and relative economic efficiency were also improved with the supplementation. Chicks fed low crude protein diet supplemented with 0.25 or 0.50% MOL have the highest dressing weight (1753 and 1738g) with no significant differences between them compared and increased serum total protein and glucose levels, while significantly decreased serum total cholesterol and aspartic transaminase (AST) enzyme compared to the other treatments with increasing the levels of MOL up to 0.50%. Morphological sections of small intestine revealed that villi height, width and height: width ratios were increased by the supplementation of different levels of MOLM (0.25 and 0.50%) but not significant except in duodenum villi width and in the ration between height and width. In general, the obtained results indicated that supplementation of 0.25% Moringa Oliefera Leaves, MOL can be used in broiler diets with low protein level to get the heaviest weight, best feed conversion, performance index, due to an increase in small intestinal absorption and activity. This, in turn, resulted in an increase in European efficiency index and economic efficiency under the experimental conditions.

Genetic Identification of <i>Tritrichomonas Foetus</i> Using Paraffin Embedded Sections of Small and Large Intestines, and Characterization of Mucosal Invasion in Piglets

Genetic Identification of <i>Tritrichomonas Foetus</i> Using Paraffin Embedded Sections of Small and Large Intestines, and Characterization of Mucosal Invasion in Piglets

Lysophosphatidic acid enhances intestinal tuft cell differentiation in Myo5b KO mice independent of Pou2f3

We have reported that adult tamoxifen‐induced, intestine‐specific, Myosin Vb knockout (Myo5b KO) mice recapitulate malabsorption syndrome in a congenital diarrheal disease, microvillus inclusion disease, and that daily treatment with lysophosphatidic acid (LPA) stimulated brush border maturation and sodium/glucose absorption in the Myo5b KO mice. Influence of Myo5b loss and LPA in sensory cell differentiation had not been studied. The present study evaluated the global changes in gene expression following Myo5b loss and LPA treatment, focusing on the differentiation of a nutrient sensor cell lineage, tuft cell. VillinCreERT2;Myo5bflox/flox mice were treated with LPA or vehicle for 4 days after a single tamoxifen injection. Immunostaining on small intestinal sections and whole‐mounted enteroids was used to quantify tuft cell density. RNA sequencing was performed on isolated jejunal epithelial cells. The sections of entire small intestine were analyzed by whole‐slide scanning and digital analysis with Cell Profiler. The frequency of DCLK1+ tuft cell was decreased by 80% with Myo5b loss, whereas enteroendocrine cells showed no difference between Myo5b KO mice and littermate controls. Paneth cell numbers were significantly increased in Myo5b KO, correlating with hyperproliferation in crypts. LPA did not alter the increased Paneth cells or Ki67+ cells. Intraperitoneal LPA treatment re‐established the frequency of tuft cells in Myo5b KO mice, indicating that a tuft cell‐specific differentiation mechanism was activated by LPA. In contrast, enteroids that were generated from VillinCreERT2;Myo5Bflox/flox jejunum had no difference in tuft cell number between hydroxytamoxifen‐treated (iKO) and control enteroids. The disrupted tuft cell differentiation by Myo5b loss was not represented in in vitro system, although microvillus formation was immature in iKO enteroids. RNA‐seq revealed that 844 genes in jejunal epithelial cells were significantly down‐regulated, and 657 genes were up‐regulated by tamoxifen‐induced loss of Myo5b compared to control. Three stem cell transcription factors were significantly increased in KO, whereas two tuft cell‐specific transcription factors (Spib and Pou2f3) were decreased. Epithelial Myo5b loss demonstrated a broad impact on epithelial cell differentiation and maturation. Treatment of Myo5b KO mice with ip LPA up‐regulated only 9 genes and had no effect on the expression of transcription factors, suggesting that LPA may modulate transporter trafficking to stimulate nutrient absorption rather than change the transcription. Myo5b loss abolished the differentiation of a sensory tuft cell, and in vivo treatment with LPA re‐established the tuft cell population independent of Pou2f3 expression. These findings suggest unexplored functions for Myo5b and LPA signaling in cell lineage choice in the intestinal epithelial cells, interacting with non‐epithelial cells.Support or Funding InformationR01 DK48370 and R01 DK70856 and a gift from the Christine Volpe Fund to J.R.G. This work was supported by core resources of the Vanderbilt Digestive Disease Center (P30 DK058404), the Vanderbilt‐Ingram Cancer Center (P30 CA68485), VUMC Digital Histology Shared Resource (supported by a VA Shared Equipment Grant 1IS1BX003097).

EFFECT OF SOME FEED ADDITIVES ON THE PERFORMANCE OF DUCKS

The present study was conducted at a private farm in Sadat City, Menoufia governorate, Egypt, throughout the experimental period from July to September 2018 in order to investigate the effect of dietary graded levels of Proact* supplementation on the Molar ducks performance, carcass traits, serum blood biochemical parameters, some histomorphological measurements and economic efficiency. A total of 240 Molar ducks one - day old chicks, were distributed at random into 8 groups each in 3 replicates, 30 chicks each. Chicks of group 1 (control) was fed the starter and finisher diets that did not supplemented with Proact. Chicks in groups 2, 3, 4, 5, 6, 7 and 8 were fed the control diets plus graded levels of Proact as follow, 0.25, 0.50, 0.75, 1.00, 1.25, 1.50 and 1.75/ kg diet, respectively. The results indicated that: 1- Groups fed the three levels (0.75, 1.00 and 0.50g / kg diet) have significantly the heaviest body weight, average daily weight gain and the best performance index compared to chicks in groups fed the highest additional level (1.75g / kg diet) and the control group. 2- Birds fed different levels of Proact significantly consumed more feed than the control group. Whereas, birds fed Proact level (1.75g/ kg diet) consumed significantly less feed than the other Proact groups and nearly equal to the control group at the same periods 0- 3, 4 – 10 and 0 – 10 weeks of age. 3- Feed conversion ratio, performance index and were significantly improved ( P ≤ 0.05) with the supplementation of Proact. 4- Addition of graded levels of Proact to Molar diets was significantly affecting all carcass traits except for pre-slaughter and dressing percentage and significantly increased both serum glucose and AST enzymes, but significantly reduced serum createnine, cholesterol, triglycerides and total lipids compared to the control group. 5- Morphological sections of Molar ducks small intestine are revealed that villi height, width and villi height : villi width are significantly increased with the Proact supplementation levels up to the level of 1.25g/ kg diet and tend to decreased after that, but still better than the control. 6- Through our inputs and outcomes for 10 weeks old Molar chick's ducks, relative economical efficiency was increased with the addition of Proact up to 0.75g/ kg diet (112%) and net revenue about 95.82 L.E compared to 75.22 L.E in the un supplemented group. 7- In general, the obtained results in the present investigation indicated that supplementation of 0.75g Proact/ kg diet can be used in diets of Molar ducks for 10 weeks to improve performance and efficiency of diet due to an increase in small intestinal absorption and activity. This, in turn, resulted in an increase in European efficiency index and economic efficiency.

Western Diet Increases the Severity of Giardia Infection in Association with Gut Microbiota Dysbiosis

AIMSGiardia duodenalis is a protozoan parasite responsible for giardiasis, a leading cause of diarrheal disease worldwide. Giardia sp. infection disrupts the host’s intestinal barrier, leading to acute, chronic, and post‐infectious gut disorders. Giardia infections are prevalent in countries with poor sanitation as well in developed countries, although disease manifestations may vary. Western diet (WD), characterized by a high intake of fat, is known to promote the development of gut inflammatory conditions. In this context, we investigated the impact of short‐term consumption of a WD on the severity and duration of giardiasis. We hypothesized that WD may contribute to the persistence of Giardia in the gut, in association with increased disease severity and microbiota dysbiosis.METHODSC57/BL6 mice were fed for two weeks with 5TJN Western high fat diet (WD) or 5TJS low‐fat (LF) diet prior to infection with Giardia duodenalis GS/M (human isolate). Gut motility and stool water content were assessed before sacrifice at day 7 post infection (PI). Small intestine and colonic sections were collected at day 7 PI for parasite burden and histopathological study. Hypoxia in the colon was assessed by injecting pimonidazole prior to euthanasia. Gut microbiota composition was determined by Illumina Mi‐Seq 16S RNA sequencing. The effect of oleic and palmitic acids (characteristic of HF diets) on Giardia trophozoite metabolic activity was determined by resazurin assay (0.1–0.25 mM). Tight junction disruption in the small intestinal cell line SCNB (dog) infected with G. duodenalis Assemblage A (NF strain) and Assemblage B (GS/M strain) in presence of oleic or palmitic acid (0.3 mM) or both was determined by immunofluorescence.RESULTSGiardia infected C57/BL6 mice fed with WD exhibited a higher trophozoite burden 7 days PI compared to LF infected mice (p<0.001). Giardia infected mice also exhibited enhanced mucosal damage (villus‐crypt ratio) when fed with WD compared to LF infected mice (p<0.01). Increased gut motility (p<0.01) and increased water stool content (p<0.05) were observed in WD infected mice. Giardia infected WD mice exhibited microbiota dysbiosis, with fecal microbiota analysis indicating an increase of Verrucomicrobia and decrease of Firmicutes. Giardia infection caused colonic mucosal normoxia only in mice given a WD. Giardia trophozoite metabolic activity was enhanced in the presence of oleic acid and palmitic acid. Oleic and palmitic acids worsened the disruption of tight junctional proteins (ZO‐1, Claudin‐1 and 4) in non‐transformed intestinal epithelial SCBN cell monolayers infected with G. duodenalis (NF and GS/M isolates).CONCLUSIONThis study gives new insight into how diet may influence the outcome of parasitic diseases. The findings suggest that short term consumption of WD may increase the severity of giardiasis by, at least in part, promoting the growth of Giardia through fatty acid supplementation, predisposing to loss of epithelial integrity, and facilitating microbiota modulation. Therefore, diet intervention such as low‐dietary fat may be beneficial during parasitic enteritis.Support or Funding InformationNSERC

Prevention of Reflux Disease After Operations On Gastric And Intestinal Tract

Creating anastomoses between the hollow organs of the abdominal cavity, abdominal formations of the retroperitoneal space and the jejunum always raises the question of preventing reflux from the jejunum into the drained cavity of the esophagus, stomach, gallbladder, external hepatic ducts, cysts of the liver and pancreas. After surgery, any reflux becomes pathological. Reflux is an obligate precancer. Thus, the reflux of bile and pancreatic juices in the stomach, the stump of the stomach and esophagus contributes to the occurrence of reflux esophagitis, reflux gastritis, ulcers and gastric cancer or its stump. After internal drainage of the cavity formation in the jejunum, postoperative reflux disease develops, which is caused by the actions of the surgeon who tried to help the patient sincerely. This allowed such states to be defined as “iatrogenic postoperative reflux disease”. The purpose of this work was to develop and introduce into practice the “plug” on the resulting loop of the jejunum, which does not migrate into the lumen of the intestine, with internal drainage of the hollow organs of the abdominal cavity and abdominal formations of the retroperitoneal space and evaluate the clinical results. As a result, the authors have developed a method for creating a “plug” on the jejunum loop, which is used for drainage, studies are being conducted on its safety, adequacy of functioning, general accessibility, and clinical situations are analyzed. For drainage of the abdominal formation impose a fistula between it and the jejunum loop 40–50 cm from the ligament of Treitz. We form an inter-intestinal fistula according to Brown, above which the length leading to a drained formation of the area of ​​the jejunum is about 10 cm, in the middle of which we impose a “plug”. The length of the small intestine section which diverts from the drained formation to the inter-intestinal brown anastomosis is about 30 cm. To form a “plug”, we use the free area of ​​the greater omentum, through which we perform a ligature of non-absorbable polypropylene material by vcol-vykola. The developed method of forming a "plug" does not cause abrupt ischemic changes in the area of ​​operation, followed by necrosis of the intestinal wall and migration of the "plug" into the intestinal lumen, and its effectiveness has been proven using clinical observations, microcirculation studies, water test results and X-ray examination. The method of creating a "stub" is promising for the internal drainage of abdominal cavity formations and retroperitoneal space, for the formation of areflux nutrient eunostoma.

Transplanted Vasoactive Intestinal Peptide-Secreting Donor Plasmacytoid Dendritic Cells Migrate to Lymphoid Organs and GvHD Target Organs

Introduction: Donor plasmacytoid dendritic cells (pDCs) have important roles in the pathogenesis of graft versus host disease (GvHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Vasoactive intestinal peptide (VIP), a potent anti-inflammatory neuropeptide with pleiotropic effects, has shown promise in managing GvHD through induction of regulatory dendritic cells (Chorny, Blood 2006) and regulatory T cells (Delgado, JLB 2005). Our previous data showed that transplantation of hematopoietic stem cells (HSC) lacking VIP expression in enhanced Th1 polarization and antiviral immunity (Li, JI 2011). Whether VIP produced by pDCs plays a key role in preventing GvHD is the focus of the current study. Method: A reductive allo-HSCT model was established by transplanting purified populations of 5 x 103 hematopoietic stem cells (HSC), 5 x 104 pDCs and 106 T cells from B6 (H-2Kb) to MHC mismatched B10.BR (H-2Kk) recipients. Mice received either VIP knockout pDCs (experimental group) or wild-type pDCs (control). Survival and clinical GvHD scores were monitored for at least 60 days. Serum pro-inflammatory cytokines were measured by luminex on day 3 and day 8. Additional recipients were transplanted for interim analyses of GvHD pathology in histological sections of small intestine and colon on day 8 and day 15. To investigate pDC migration and proliferation status, 5 x 104 luciferase-expressing pDCs, together with 5 x 103 WT B6 HSC, were transplanted to MHC mismatched Balb/c (H2kd) recipients. Bioluminescent imaging was performed on day 14. We used mice carrying an eGFP reporter linked to a VIP promoter to measure VIP production by donor cells in vivo. The construct has the eGFP reporter gene, followed by a polyadenylation sequence, inserted into BAC clone RP23-25A8 at the initiating ATG codon of the first coding exon of the VIP gene, so that eGFP expression is driven by the regulatory sequences of the BAC gene. The BAC is inserted randomly in the genome. We used serial breeding to establish the eGFP-VIP reporter in B6 mice. Bone marrow pDCs were sorted from transgenic eGFP-VIP B6 mice and used in transplant experiments, allowing VIP gene activity from donor-derived pDCs to be detected as GFP expression using confocal microscopy. Results: Compared to recipients of WT pDCs, recipients of VIP-KO pDCs have a significantly lower survival rate with increased GvHD clinical score (Figure 1; *p<0.05). Transplant recipients of VIP-KO donor pDC had higher serum pro-inflammatory cytokine levels on day 3 and day 8 respectively. (Figure 2). At 2 weeks post-transplant donor pDCs migrated primarily to the spleen, with limited presence of donor pDCs in the gastrointestinal tract and lung (Figure 3). Compared to mice transplanted with WT pDCs, recipients of VIP-KO pDCs had increased donor T cell migration to the lung and paw, which are chronic GvHD target organs after two weeks. (Figure 4; *p<0.05). When stimulated with IFN-g, eGFP-VIP transgenic pDCs have activation of the VIP gene promoter in vitro, as detected using eGFP (Figure 5). Confocal microscopy data showed that pDCs have activated the VIP promoter and secreted more VIP following stimulation by IFN-g, consistent with VIP up-regulation by the inflammatory mileu in allo-transplant recipients. Conclusions: These data show that lack of VIP secretion by donor pDCs resulted in lower survival rates and higher GvHD scores in allo-BMT recipients. Recipients of VIP KO donor pDC had increased donor T cell proliferation in GvHD target organs and increased GvHD clinical signs. The major site of donor pDCs migration in recipients is spleen, suggesting that pDCs may interact with other lymphoid cells, such as T cells, inducing a more tolerogenic environment. Donor pDCs were also found, to a lesser extent, in the lung and gastrointestinal tract, which are both GvHD target organs. Thus, pDCs may protect recipients from GvHD by limiting the activity of alloreactive T cells at these sites. The mechanism by which VIP-secreting donor pDCs regulate GvHD will be discussed. Disclosures Waller: Amgen: Consultancy; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Pharmacyclics: Other: Travel expenses, Research Funding; Cerus Corporation: Other: Stock, Patents & Royalties; Chimerix: Other: Stock; Cambium Oncology: Patents & Royalties: Patents, royalties or other intellectual property ; Kalytera: Consultancy.

Effect of bolus viscosity on carbohydrate digestion and glucose absorption processes: An in vitro study

Digestion is the process of breaking down food into smaller nutrient components which can be easily absorbed in the intestinal tract. The aim of this study was to experimentally investigate the influence of bolus (gastric content) viscosity on digestion and nutrient absorption processes, using an in vitro gastrointestinal model, the TIM-1 system. Two types of simple carbohydrates, namely, glucose and maltodextrin, were used as model foods. The initial bolus viscosity was varied (∼1 mPa·s, ∼15 mPa·s, and ∼100 mPa·s) using different glycerol-water proportions. A fluorescent molecular rotor compound (Fast Green For Coloring Food) was used to monitor viscosity changing patterns of the gastrointestinal content during digestion in the in vitro stomach and small intestinal sections. The digested-nutrient absorption data indicated that the initial bolus viscosity did not significantly affect the glucose absorption process in the small intestine. However, an increase in the initial bolus viscosity from ∼1 mPa·s to ∼15 mPa·s reduced the maltodextrin to glucose conversion by 35%. A further increase in the initial bolus viscosity from ∼15 mPa·s to ∼100 mPa·s did not significantly reduce the maltodextrin to glucose conversion.

Therapeutic effect of P-coumaric acid in the experimentally infected rats with Salmonella typhi

Salmonella enterica subspecies enteric serotype typhi which infect the human and cause typhoid fevers (systemic infection). Experimental infection with Salmonella typhi are very important because they explains the pathogenicity of Salmonella typhi through a histopathological findings. The herbal inhibitors like P-Coumaric acid explain anti-inflammatory effect and have a selective effect against type III secretion system formation of Salmonella typhi. our study includes 15 adult male albino rats were divided into three groups each group composed from 5 rats. A group: were orally challenged with 1ml of 3X106 CFU of Salmonella typhi for one day. Group B: were also orally administered with 1ml of 3X106 CFU of Salmonella typhi, additionally, they also orally received 5ml of 100mg/kg of P-Coumaric acid for three consecutive days. Group C: serving as control group, they orally administered with 5ml of normal saline. All the animals are sacrificed after five days post infection, liver, small intestine and colon specimens were taken for histopathological investigation. the histological sections of liver and small intestine of group A animals showed pathological changes like degeneration of hepatic cells and inflammatory cell infiltration of the lamina properia in small intestine, while the group B animals showed normal liver and small intestine sections like the control group

Slow cycling intestinal stem cell and Paneth cell responses to Trichinella spiralis infection

There is limited information regarding responses by slow cycling stem cells during T. spiralis-induced T-cell mediated intestinal inflammation and how such responses may relate to those of Paneth cells. Transgenic mice, in which doxycycline induces expression of histone 2B (H2B)-green fluorescent protein (GFP), were used. Following discontinuation of doxycycline ("chase" period), retention of H2B-GFP enabled the identification of slow cycling stem cells and long-lived Paneth cells. Inflammation in the small intestine (SI) was induced by oral administration of T. spiralis muscle larvae. Epithelial retention of H2B-GFP per crypt cell position (cp) was studied following immunohistochemistry and using the Score and Wincrypts program. Compared to non-infected controls, there was significant reduction in the number of H2B-GFP-retaining stem cells in T. spiralis-infected small intestines. H2B-GFP-retaining stem cells peaked at around cp 4 in control sections, but smaller peaks at higher cell positions (>10) were seen in sections of inflamed small intestines. In the latter, there was a significant increase in the total number of Paneth cells, with significant reduction in H2B-GFP-retaining Paneth cells, but a marked increase in unlabelled (H2B-GFP-negative) Paneth cells. In conclusion, following T. spiralis-infection, putative slow cycling stem cell numbers were reduced. A marked increase in newly generated Paneth cells at the crypt base led to higher cell positions of the remaining slow cycling stem cells.

Grape-Seed Proanthocyanidins are Able to Reverse Intestinal Dysfunction and Metabolic Endotoxemia Induced by a Cafeteria Diet in Wistar Rats

We evaluated the effectiveness of pharmacological doses of grape-seed proanthocyanidin extract (GSPE) in reversing intestinal barrier alterations and local inflammation in female Wistar rats fed a long-term obesogenic diet. Animals were fed a 17-week cafeteria diet (CAF diet), supplemented with daily GSPE doses (100 or 500 mg kg−1 body weight) during the final two weeks. CAF diet enhanced the intestinal permeation of an orally administered marker (ovalbumin, OVA) and increased the plasma levels of tumor necrosis factor-α (TNF-α) and lipopolysaccharides (LPS) in 2–3-fold. Ex vivo Ussing chamber assays showed a 55–70% reduction in transepithelial electrical resistance (TEER) and increased the TNF-α secretions in both small and large intestinal sections with a 25-fold increment in the ileum. Ileal tissues also presented a 4-fold increase of myeloperoxidase (MPO) activity. Both GSPE-treatments were able to restitute TEER values in the ileum and colon and to reduce plasma LPS to basal levels without a dose-dependent effect. However, effects on the OVA permeation and TNF-α secretion were dose and section-specific. GSPE also reduced ileal MPO activity and upregulated claudin 1 gene expression. This study provides evidence of the efficacy of GSPE-supplementation ameliorating diet-induced intestinal dysfunction and metabolic endotoxemia when administered at the end of a long-term obesogenic diet.

Dimethyl Oxalyl Glycine a Prolyl Hydroxylase Inhibitor Decreases Early Gastrointestinal Gvhd Via Two Independent Pathways of Apoptosis after Allogeneic Hematopoietic Cell Transplantation

Abstract Introduction: Allogeneic hematopoietic cell transplantation (HCT) is increasingly utilized in the treatment of hematologic malignant and non-malignant diseases. Therapy-related toxicity and graft versus host disease (GVHD) remain major challenges as they significantly contribute to treatment related morbidity and mortality. Prolyl hydroxylase inhibitors (PHI) interfere with signaling cascades of inflammation and cell death. Their beneficial use in experimental models of ulcerative colitis and lung allograft rejection resulted in the hypothesis, that use of the PHI dimethyl oxalyl glycine (DMOG) reduces GVHD after murine allogeneic HCT. We already showed that DMOG treatment of allogeneic recipients resulted in significantly improved survival by day +50 compared to allogeneic controls, and was associated with significant decrease in organ pathology and proinflammatory cytokines (data not shown). We now expanded our study to better understand the underlying mechanisms for this observation. Methods: Acute GVHD was induced in lethally irradiated BALB/c mice followed by the infusion of 5 million bone marrow cells mixed with 4 million splenocytes from allogeneic C57BL/6 donors. DMOG dissolved in PBS was given at 8mg per mice intraperitoneally while controls received PBS only. Animals were monitored for clinical GVHD and survival and analyzed at day +7 and at day +50 post HCT. Formalin-fixed paraffin sections were processed for immunohistochemistry with CD3, Hif-1 alpha, or FADD pSer191 antibodies. Western blot analysis was performed after CD3/CD28 antibody activation of T cells to see the effect of DMOG treatment on Hif-1 α expression. Effect of DMOG on apoptosis and CD137 expression was analyzed by flow cytometry. Results: Our results demonstrate that the protective effect of DMOG is mainly caused by two independent pathways of apoptosis. DMOG treatment of allogeneic recipients resulted in decreased T cells infiltration (p=0.0534) and (Fas associated death domain) FADDpSer191 level (p<0.05) in small intestine sections at day +50. Decreased FADD pSer191 indicates decreased TNF-a induced epithelial cell apoptosis. This was associated with increased Hif1- α expression at day+7 (p<0.05) as well day+50 (p<0.05) as shown by histochemical analysis of small intestine sections. DMOG treatment of activated T cells showed increased expression of CD137 (p<0.005) and Annexin V FITC (p<0.005) staining suggesting that DMOG subjects T cells to activation induced cell death (AICD) via induced CD137 expression. Microscopic scanning of DMOG treated T cells showed condensed chromatin along with numerous fragmented nuclei, indicative of apoptotic cell. In vitro proliferative T cell responses were suppressed (p<0.05) in the presence of DMOG in a dose-dependent manner along with increase in Hif1- α level (p<0.05) as analyzed by western blot. Conclusion: Our data demonstrates protective effects of DMOG on the GI tract early after allogeneic HCT which is associated with improved survival and reduced lung injury at later time point. DMOG treatment results in the induction of CD137 on T cells subjecting them to AICD and represses FADD level in epithelial cells regulating their apoptosis. Taken together, DMOG treatment reduces GVHD via a two pronged mechanism, possibly allowing for novel strategies to prevent and treat this deleterious disease. Disclosures Palaniyandi: Pharmacyclics: Research Funding; Takeda Pharmaceutical International, Inc.: Research Funding; JAZZ: Research Funding.

Abstract 16947: Mitochondrial Calcium Uniporter Channel Blockade Ameliorates Sepsis Induced Cardiomyopathy

Introduction: Mitochondria serve as critical Ca 2+ sinks, regulating the duration and magnitude of c Ca2+ transients and c Ca2+ entry into the mitochondrion. Mitochondrial calcium uniporter (MCU) channel is activated in conditions of inflammation and oxidative stress.MCU and its activity are potential therapeutic targets for cardiac hypertrophy and heart failure. Hypothesis: Blunting of mCa2+ uptake can contribute to increased cCa2+ transients associated with excitation-contraction coupling and offer cardio protection following sepsis induced stress. Methods: MCU C96A KI generated using CRISPR-Cas9 d10 nickase mRNA co-injected with gRNA into wild-type(C57BL6) zygotes and then implanted into pseudo-pregnant female mice.To elucidate mechanistic role of mCa2+ in modulating cardiac contractility and myocardial bioenergetics, adult cardiomyocytes were isolated from mouse model and subjected to hypoxia/re-oxygenation injury.Polymicrobial Sepsis in 6-week-old C57BL/6J mice were created using Cecal ligation and puncture (CLP) and MCU oxidation at 1, 6, 12 and 24 hours post injury was compared with Sham and control mice (n=15). MCU protein quantification was done with western blot, functional assessment of Reactive oxygen species (ROS) by confocal laser microscopy, echocardiography with Vevo2100 imaging system. Mice were pretreated with MCU blocking agent Ru360 1 hr prior to CLP and compared with the control group at 6 hours. Results: Cardiomyocytes exposed to hypoxia/re-oxygenation(h/R ) stress exhibited increased MitoSOX red fluorescence.MCU oxidation was ubiquitously induced in all tissues post CLP surgery and was most significant at 6 hrs (Fig 1a). Oxidation of MCU was marked by increase in ROS in tissue sections of small Intestine of CLP versus the Sham group(p<0.001). (Fig 1 b &c). Pretreatment with Ru360 ameliorated MCU oxidation at 6 hours post CLP injury. MCUC96AKI had significantly decreased ejection fraction and fractional shortening compared to the wild type mice. Further the survival in MCU C96A KI was significantly better than the WT or MCU KO mice ( p<0.05). Conclusions: Inhibition of mitochondrial calcium uniporter is a potential therapeutic target and protects from sepsis induced cardiomyopathy by mitigating mitophagy.

Growth Performance and the Intestine Morphometry of Japanese Quail That Fed Diets Supplemented With Vitamin E and Selenium

Background: Small intestine and peripheral organs are the main organs for digestion and absorption in the gastrointestinal tract. OBJECTIVES: To investigate effects of different levels of Vitamin E and Selenium on growth performance and small intestine morphometry in Japanese quail. METHODS: A total of 405 mixed sex Japanese quails were randomly allocated into 9 experimental groups by 3×3 factorial design (3 replications and 15 chicken per pen) using Vitamin E (0, 150 and 300 mg/kg) and sodium Selenium (0, 0.2 and 0.4 mg/kg feed) for the entire study. Then, feed intake, body weight gain and food conversion ratio were recorded on days 17-21, 22-35 and 17-35 of the study. At day 35 of the study, 3 birds were randomly selected from each replication, slaughtered and various sections of small intestine (10, 50 and 90% of small intestine length) sampled for morphometry characteristics. Villi height and crypts depth were measured microscopically. RESULTS: According to the results, different levels of Vitamin E and Selenium supplements had no effect on growth performance (P>0.05). Also, supplementation of diet with Vitamin E and Selenium significantly increased villi height and crypts depth in various sections of small intestine on day 35 (P<0.05). CONCLUSIONS: These results suggest Vitamin E and Selenium supplemented diets had beneficial effect on small intestine morphometry characteristics in Japanese quail. Keywords:

Morphological, Mechanical and Mucoadhesive Properties of Electrospun Chitosan/Phospholipid Hybrid Nanofibers.

This study aimed to develop hybrid electrospun chitosan–phospholipid nanofibers and investigate the effect of phospholipid (P) content and chitosans (Ch) molecular weights (Mw) and degree of acetylation (DA), on the morphological, mechanical and mucoadhesive properties of the nanofibers. Electrospun Ch/P nanofibers exhibited a smooth and uniform surface with average diameters ranging from 300 to 1000 nm, as observed by scanning electron microscopy (SEM). The average diameter of the nanofibers was observed to increase with the increase of the Mw and degree of deacetylation of Ch, and phospholipid content. The elastic and adhesive properties of the nanofibers were determined by atomic force microscopy, and displayed higher values for higher Mw and lower DA Ch used. The elastic modulus of electrospun Ch/P hybrid fibers determined for the different conditions tested was found to be in the range of 500 and 1400 MPa. Furthermore, electrospun Ch/P nanofibers displayed mucoadhesive properties expressed by the work of adhesion calculated after the compression of the nanofibers against a section of pig small intestine. Our results showed that the increase in phospholipid content and DA of Ch decrease the work of adhesion, while the increase of Mw resulted in slightly higher work of adhesion of the nanofibers.

Human evolutionary loss of epithelial Neu5Gc expression and species-specific susceptibility to cholera.

While infectious agents have typical host preferences, the noninvasive enteric bacterium Vibrio cholerae is remarkable for its ability to survive in many environments, yet cause diarrheal disease (cholera) only in humans. One key V. cholerae virulence factor is its neuraminidase (VcN), which releases host intestinal epithelial sialic acids as a nutrition source and simultaneously remodels intestinal polysialylated gangliosides into monosialoganglioside GM1. GM1 is the optimal binding target for the B subunit of a second virulence factor, the AB5 cholera toxin (Ctx). This coordinated process delivers the CtxA subunit into host epithelia, triggering fluid loss via cAMP-mediated activation of anion secretion and inhibition of electroneutral NaCl absorption. We hypothesized that human-specific and human-universal evolutionary loss of the sialic acid N-glycolylneuraminic acid (Neu5Gc) and the consequent excess of N-acetylneuraminic acid (Neu5Ac) contributes to specificity at one or more steps in pathogenesis. Indeed, VcN was less efficient in releasing Neu5Gc than Neu5Ac. We show enhanced binding of Ctx to sections of small intestine and isolated polysialogangliosides from human-like Neu5Gc-deficient Cmah-/- mice compared to wild-type, suggesting that Neu5Gc impeded generation of the GM1 target. Human epithelial cells artificially expressing Neu5Gc were also less susceptible to Ctx binding and CtxA intoxication following VcN treatment. Finally, we found increased fluid secretion into loops of Cmah-/- mouse small intestine injected with Ctx, indicating an additional direct effect on ion transport. Thus, V. cholerae evolved into a human-specific pathogen partly by adapting to the human evolutionary loss of Neu5Gc, optimizing multiple steps in cholera pathogenesis.

Re-evaluation of endogenous development of Eimeria bareillyi Gill, Chhabra and Lall, 1963 in water buffalo (Bubalus bubalis).

Water buffalo (Bubalus bubalis) is important for the economy of Asia, South America and parts of Europe. Coccidiosis is an important cause of neonatal mortality in livestock, including buffalo. Of more than 12 species of Eimeria in buffalo, Eimeria bareillyi is the most pathogenic. There are uncertainties concerning its asexual and sexual development. During a previously reported outbreak of fatal enteritis associated with E. bareillyi in buffaloes in the Netherlands, sections of small intestine were re-evaluated histologically and by transmission electron microscopy (TEM) to seek details of endogenous development. Profuse asexual multiplication occurred in the jejunum and ileum. Light microscopic examination revealed that parasites divided in two (probably endodyogeny) or more organisms. There were two or more generations of morphologically different merozoites; some of these observations were confirmed by TEM. Details of gametogonic development, including oocyst wall formation are provided. Schizogonic and gametogonic development described in the present study can serve as a guide for differential diagnosis of Eimeria species in histological sections of intestines of buffaloes.