Gastrointestinal Histology Research Articles

Gastrointestinal Histology in Knoack Out Mouse Models and Temporal Induction of CFTR Protein Function in Mice

Gastrointestinal Histology in Knoack Out Mouse Models and Temporal Induction of CFTR Protein Function in Mice

Role of Fractionation in Local Control of Spinal Metastases Treated with Stereotactic Radiosurgery

Optimal fractionation of spinal stereotactic radiosurgery (SRS) for spine metastases remains unknown. Retrospective data suggest decreased local failure (LF) with fractionated SRS of brain metastases. We evaluated our institutional outcomes of spinal SRS with the hypothesis that fractionation improves the rate of local failure compared to single-fraction treatment. This IRB-approved, retrospective analysis included patients with spine metastases treated with spinal SRS between October 2002 and November 2014 with evaluable follow-up imaging and no prior irradiation to the given segment. The exposure of interest was single- or multi-fraction SRS with a primary endpoint of the cumulative incidence of LF with death as a competing risk. We assessed bivariate associations between fractionation and single-fraction equivalent dose (SFED in Gy10) as well as high-risk features, defined as epidural extension (Bilsky Scale), paraspinous extension, and gastrointestinal (GI) vs non-GI primary. We calculated the rates of LF and vertebral body compression fracture (VCF) at 1-year, and assessed LF by fractionation when limited only to courses receiving SFED>18 Gy. We analyzed the association between fractionation and LF using subdistribution hazard ratios (SHR) estimated from competing risks regression with death as a competing risk and adjusting for lesion-specific characteristics as well as SFED to determine contribution of these variables to the estimated effect of fraction number on LF. We calculated relative attenuation for the contribution of SFED to this association, defined as [SHRfractions-SHRfractions+SFED] ÷ [SHRSFED-1]. In 293 patients with 516 spinal segments, lesions treated with single fraction compared to multi-fraction SRS had less epidural (19% vs 36%, p<0.001) and paraspinous (20% vs 35%, p<0.001) extension, more GI histology (17% vs 10%, p = 0.039), received a higher mean SFED (18.3 Gy vs. 16.6 Gy, p<0.001), and had a lower 1-year LF (8% vs 14%, p = 0.02), with no difference in VCF (7% vs. 5%, p = 0.38). After adjusting for high-risk features, single fraction SRS was associated with lower LF (SHR = 0.45, 95% CI 0.24-0.84, p = 0.02). After adjustment for SFED, this association of fractionation was attenuated by 53% and became insignificant (SHR = 0.78, 95% CI 0.44-1.37, p = 0.38). Overall, 1-year LF for SFED>18 Gy was 6% compared to 15% for <18 Gy (p<0.001). When limited to courses with SFED>18 Gy (n = 261), single fraction SRS had no improvement in 1-year LF compared to multi-fraction (6.6% vs 4.6%, p = 0.77). Single fraction SRS was associated with better local control compared to multi-fraction; however, much of this association was attenuated by SFED but not by high-risk features of treated lesions. To clarify the role of fractionation, we have initiated a prospective, randomized trial of single vs. multi-fraction SRS utilizing the same SFED.

Evaluation of probiotic potentiality of GM-Lac (Lactobacillus and Bifidobacterium) in juvenile Asian seabass Lates calcarifer

This study was conducted to evaluate the probiotic potentiality of GM-Lac (a mixture that comprises four lactic acid bacteria (LAB) strains– Limosilactobacillus fermentum GMNL-93, Lactiplantibacillus plantarum GMNL-141, Lacticaseibacillus casei GMNL-277, and Bifidobacterium lactis GMNL-550) applied in fish aquaculture. First, the four LAB isolates (GMNL-93, −141, −277, and −550) constituting GM-Lac were tested individually for antimicrobial activity in vitro. Then, GM-Lac was incorporated in fish feed at a dose of 5 × 106 cfu (g feed)−1 and fed to Asian seabass (Lates calcarifer) juveniles for eight weeks to assess the effects on growth performance, gastrointestinal (GI) histology and microflora, muscle amino acid composition, and disease resistance. According to the result of the zone of inhibition test, each GMNL strain presented noticeable antimicrobial activity against bacterial pathogens common in aquaculture. After the 8-week feeding trial, the fish in the GM-Lac group had higher final weight (p < 0.05), percent weight gain, specific growth rate, and feed intake than the fish in the Ctrl group, along with higher microvillus length in GI tract (p < 0.05) and greater total amino acid concentration in muscle. Furthermore, the relative abundance of microbial phyla in the GI tract of the fish fed with GM-Lac changed significantly, accompanied by increasing probiotic bacteria and decreasing pathogenic bacteria in the intestine and pyloric caeca. In the challenge test, the fish receiving GM-Lac all survived without any clinical symptoms when challenged with Streptococcus iniae. This study concludes that GM-Lac (the combination of L. fermentum GMNL-93, L. plantarum GMNL-141, L. casei GMNL-277, and B. lactis GMNL-550) is recommended as a probiotic supplement with high potential in aquaculture practice, which has antimicrobial activities against aquatic pathogens and benefits in enhancing growth performance, intestinal health, muscle amino acid composition, and disease resistance in juvenile Asian seabass.

Gastroprotective activity of yogurt fortified with purple roselle extract in rats exposed with 2,3,7,8-TCDD

Persistent organic pollutant 2,3,7,8-Tetrachlorodiobenzo-p-dioxin (TCDD), known as harmful congeners of dioxin, has many adverse effects on human or animal health. This TCDD enters the body through inhalation, ingestion, and skin contact. Purple Roselle is a well-known herb-medicinal plant having antioxidant properties. This study aimed to evaluate the antioxidant capacity of purple roselle water extract along yogurt against dioxin exposure. The antioxidant properties of the extract were measured by malondialdehyde (MDA) levels and gastrointestinal histology. For this, 25 white male rats (Rattus norvegicus) were used, and these rats were divided into five groups negative control, positive control (TCDD 200 ng/kg BW), and three treatment groups (TCDD 200 ng/kg BW + yogurt fortified with purple roselle water extract concentrations 0.5, 1, and 1.5 percent), all the treatments were given orally. Gastric MDA was determined quantitatively using the Thiobarbituric Acid (TBA) method and the one-way ANOVA test, continued by a Tukey post hoc test with a confidence level of 95% while the gastric histology was observed descriptively. Results of the study revealed that supplementation of fortified yogurt with 1% purple roselle extract could dramatically reduce MDA levels (p&lt;0,05) and heal histological damage in the lamina propria mucosa of stomach rats subjected to TCDD. Results of the study can be concluded that consuming yogurt with purple roselle extract can reduce MDA levels and repair histological damage to the gastric mucosa caused by dioxin exposure.

Effect of chronic heat stress on gastrointestinal histology and expression of feed intake-regulatory hormones in broiler chickens

Heat stress (HS) dramatically impairs the growth performance of broiler chickens, mainly as a consequence of reduced feed intake due to the loss of appetite. This study was aimed at evaluating the alterations induced by chronic HS conditions on the morphological and morphometric features of the gastrointestinal (GI) tract and on the expression of some enteroendocrine cells (EECs) involved in the regulation of feed intake in chickens. Three hundred male chickens (Ross 308) were divided into two experimental groups and raised either in thermoneutral environment for the whole fattening period (0–41 days) (TNT group) or subjected to chronic HS conditions (30 °C for 24 h/day) from 35 to 41 days (HS group). Samples of proventriculus, duodenum, jejunum and cecum were collected from 24 broilers (12/group). Haematoxylin-eosin was used for the morphometric evaluations, while immunohistochemistry was applied for the evaluation of EECs expressing ghrelin (GHR), cholecystokinin (CCK), neuropeptide Y (NPY), glucagon-like peptide-1 (GLP-1), and serotonin (5-HT). In the proventriculus, HS reduced total wall thickness and mucous layer height (P ≤ 0.01) as well as mean diameter, circumference, and area of the compound tubular glands (P ≤ 0.001) with respect to TNT. The small intestine of HS birds was characterised by decreased villous height and total thickness (duodenum, P ≤ 0.01; jejunum, P ≤ 0.001), whereas crypt depth and width were reduced only in the jejunum (P ≤ 0.01). HS had negligible effects on the morphological aspects of the cecum. In the proventriculus, an increase in GHR and NPY EECs was observed in response to HS (P ≤ 0.001). Similarly, the small intestine villi of the HS group showed greater GLP-1 (P ≤ 0.05), 5-HT (P ≤ 0.001) and CCK (P ≤ 0.01) EECs. Moreover, the expression of 5-HT EECs was higher in the duodenal (P ≤ 0.01) and jejunal (P ≤ 0.01) crypts of HS birds, whereas GLP-1 and CCK EECs increased only in jejunal crypts (P ≤ 0.05). Finally, 5-HT EEC expression was increased in the cecum of HS group (P ≤ 0.01). In conclusion, these outcomes demonstrate that chronic HS induces morphometric alterations not only in the small intestine but also in a key organ such as the proventriculus. Furthermore, HS conditions affect the presence and distribution of EECs, suggesting that some GI peptides and biogenic amine may be implicated in the regulation of appetite and voluntary feed intake in heat-stressed broiler chickens.

Dietary astaxanthin modulated the performance, gastrointestinal histology, and antioxidant and immune responses and enhanced the resistance of Litopenaeus vannamei against Vibrio harveyi infection

Astaxanthin (AX) is one of the commonly used feed supplements to enhance the growth performance and provide antioxidant and immune functions of several aquatic animals. In the current study, juveniles of white-leg shrimp (Litopenaeus vannamei) with mean initial weight of 0.340 ± 0.041 g were fed with diets supplemented with 0 (control), 25, 50, 100, and 200 mg/kg feed for 8 weeks. At the end of the feeding trial, shrimps were exposed to Vibrio harveyi, and their mortality rates were observed for additional 10 days. The growth indices in the AX-fed groups were significantly (P < 0.05) higher than what were observed in shrimps in the control group. Dietary AX stimulated the final weight, weight gain, and specific growth rate and optimum growth levels were achieved at 100–200 mg AX/kg feed. Furthermore, the AX-enriched diets significantly enhanced feed intake more than the control diet, and the amount of AX had no effects on feed conversion ratios. In comparison to the control group, the AX-fed animals had significantly (P < 0.05) higher villi length, villi width, and absorption area and their optimum values were observed at 100–200 mg AX/kg feed treatments. Moreover, the intestinal morphometry especially villi and its crypt, both internal and external tunica muscularis, and submucosal tissues did not show any inflammatory and/or degenerative changes in AX-fed shrimp. Furthermore, the dietary AX at escalating levels linearly and quadratically enhanced (P < 0.05) the activities of serum superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and increased levels of total antioxidant capacity and reduced glutathione. In addition, malondialdehyde levels decreased significantly in AX-fed animals, and the highest levels were observed in the control group (without AX). The expression levels of cMn-SOD, CAT, and GPx genes were significantly upregulated in the hepatopancreas of L. vannamei fed with AX-enriched diets (especially in the 200-mg/kg feed treatment) as compared with the control diet. The immunity indices of the AX treatments (hematocyte count, total protein, lysozyme, phagocytic activity, and phenoloxidase) of L. vannamei were linearly (P < 0.05) and quadratically (P < 0.05) increased. This study revealed the antibacterial activity of AX against V. harveyi abundance. After the bacterial challenge, feeding L. vannamei with dietary AX significantly increased (P < 0.05) the relative percentage of survival, especially in the treatment of 200-mg/kg diet (82.7%). The findings of the current study demonstrate that dietary AX (100–200 mg/kg feed) was effective in enhancing the growth, antioxidant status, immune response, and increasing the resistance of L. vannamei against V. harveyi infection.

The gastrointestinal tract histology of Eliomys quercinus and Glis glis

The Garden Dormouse (Eliomys quercinus) and the Edible Dormouse (Glis glis) differ in their omnivorous diet, with the first being more carnivorous and readily consuming small vertebrates and even poisonous invertebrates, while the latter consumes more plant-based foods. These differences could lead to histological differences in their gastrointestinal tract, which we wanted to test. Six animals of each species were caught in nature and fed the same diet for two weeks to diminish the impact of the short-term external factors. Tissue samples were taken of middle oesophagus, glandular stomach, middle small intestines and large intestines. They were stained with H&amp;amp;E for measuring the thickness of layers in oesophagus gastric mucosa and epithelial cell height in the intestines. Alcian blue - PAS stain was used for goblet cell count and type identification in the intestines, differentiating blue (acidic), lilac (neutral) and purple (mixed) cells. Results showed Eliomys quercinus having greater variance in oesophageal layer thicknesses, significantly thicker muscularis mucosae and thinner tunica muscularis in the oesophagus, and thicker gastric mucosa compared to Glis glis. Eliomys quercinus also had a significantly higher total goblet cell count in both intestines and a higher count of each type in the large intestine, although Glis glis had far more mixed and neutral goblet cells in the small intestine. Both showed intriguingly low numbers of neutral goblet cells in the small intestine. These results indicate interesting differences in gastrointestinal histology and goblet cell distribution in comparison with their distinct diets.

Could Dietary Supplementation with Different Sources of N-3 Polyunsaturated Fatty Acids Modify the Rabbit Gut Microbiota?

The present study evaluated the effects of feed supplemented with two dietary sources of n-3 polyunsaturated fatty acids (PUFAs; fish oil and extruded flaxseed) on the gut microbiota, caecal fermentations, gastrointestinal histology, and histochemistry in rabbits. Fifteen male New Zealand White rabbits were divided into three groups (n = 5/group) and fed with different diets from weaning (35 days of age) until slaughtering (90 days of age): C group, fed with a commercial diet; F group, supplemented with 10% of extruded flaxseed; and O group, supplemented with 3.5% of fish oil. At slaughter, the content of the stomach, duodenum, jejunum, ileum, caecum, and colon was collected and analyzed by Next Generation 16S rRNA gene sequencing. Tissue samples of the same tracts were evaluated with histological and histochemical analysis. Ammonia and lactic acid in the caecum were also quantified. Twenty-nine operational taxonomic units (OTUs) were significantly different between groups. Groups receiving n-3 PUFAs supplementation showed an increase in Bacteroidetes and Lachnospiraceae in several gastrointestinal tracts, while Bacilli abundance, as well as Firmicutes/Bacteroidetes ratio, were reduced compared to the control group (for all p < 0.05). Caecal ammonia was lower in the F than C group (p < 0.032), whereas no difference was found for lactic acid. Finally, histological evaluations revealed a mild hemorrhagic infiltration and vessels ectasia in the stomach mucosa of both F and O groups, but no effect of nutritional treatment was evidenced by the histochemical analyses. In conclusion, n-3 PUFAs supplementation could modify the rabbit gut microbiota and fermentation. The increase in beneficial bacterial populations may, at least partially, explain the positive effects of n-3 PUFAs diet supplementation on human and animals’ health, although the appropriate dosage should be established.

Significance of granulomas in the outcomes of Crohn's disease patients.

BackgroundThe presence of granulomas in the gastrointestinal (GI) tract is one of the characteristic histologic features of Crohn’s disease (CD). The clinical significance of granulomas remains unclear. In this study, we aimed to determine whether the presence of granulomas on endoscopic pinch biopsy or surgical resection from the upper or lower GI tract is associated with worse outcomes among patients with CD.MethodsThis was a retrospective chart review of patients with CD evaluated at a tertiary care center between 1996 and 2019. Patients were divided into 2 groups based on the presence or absence of granulomas on GI histology. Clinical and laboratory data, and outcomes of interest, were obtained from the electronic medical records. Patients’ characteristics and outcomes were compared between the 2 groups.ResultsA total of 237 patients were included in our study; 41 (17.3%) had granulomas on their biopsy/resection specimen. The presence of granulomas in the GI tract was significantly associated with the development of intra-abdominal abscesses and/or fistulas (P=0.037), greater utilization of immunomodulators (P=0.029), and greater use of immunosuppressive medications (immunomodulator and/or biologic therapy) (P=0.015). No significant differences were found between the 2 groups in terms of number of hospitalizations, presence of perianal disease, intestinal resection, mean age, mean age at initial diagnosis of CD, duration of disease, sex, or smoking history.ConclusionsThe presence of granulomas in the GI tract of CD patients may serve as a prognostic biomarker of worse disease severity. Larger studies are needed to better validate this finding.

Dietary fermented rapeseed or/and soybean meal additives on performance and intestinal health of piglets

The aim of the study was to assess the effect of fermented dried soybean (FSBM) and/or fermented rapeseed meal (FRSM) in diets for weaned piglets on production results, nutrient digestibility, gastrointestinal tract histology, and the composition of the gut microbiota. Piglets in the control group received standard diets with soybean meal. Animals in all experimental groups received diets in which a portion of the soybean meal was replaced: in group FR—8% FRSM; in group FR/FS—6% FRSM and 2% FSBM; in group FS/FR—2% FRSM and 6% FSBM and in group FS—8% FSBM. The use of 8% FRSM or 6% FRSM and 2% FSBM in the piglet diets had a positive effect on average daily gains. Piglets from the FR and FR/FS groups had the highest feed conversion rate. Group FS/FR and FS piglets had significantly lower mortality and lower incidence of diarrhoea. Piglets fed a diet with the fermented components, in particular with 8% FRSM or 6% FRSM and 2% FSBM, exhibited a positive effect on the microbiological composition and histology of intestines, which resulted in improved nutrient digestibility coefficients (ATTD and AID).

Comparison of Nonesophageal Eosinophilic Gastrointestinal Disorders with Eosinophilic Esophagitis: A Nationwide Survey

Eosinophilic esophagitis (EoE) has increased rapidly and has been well characterized. However, no nationwide survey has been conducted regarding non-esophageal eosinophilic gastrointestinal disorders (non-EoE EGIDs), and they remain poorly understood. To compare the clinical features and natural histories of non-EoE EGIDs and EoE by using the same questionnaire, for all ages. We conducted a nationwide hospital-based survey of patients who visited hospitals from January 2013 through December 2017. We randomly selected 10,000 hospitals that perform endoscopy. We analyzed the demographics, symptoms, gastrointestinal histology, treatments, and natural histories of EoE and non-EoE EGIDs. A total of 2906 hospitals responded to the questionnaire. We identified 1542 patients and obtained detailed data for 786 patients, consisting of 39% EoE and 61% non-EoE EGIDs. The clinical characteristics were analyzed for patients who met the "definite" criteria that excluded comorbidities. Non-EoE EGIDs showed no gender difference, whereas EoE was male-predominant. Tissue eosinophilia was often seen in the small intestine (62%) and stomach (49%). The frequency of hypoproteinemia was high (27%) in childhood. Children also had more serious symptoms and complications than adults: restriction of daily life activity (P= .009), failure to grow/weight loss (P= .008), and surgery (P= .01). For both diseases, the most common natural history was the continuous type: 66% (95% confidence interval [CI]: 58-74) in EoE and 64% (95% CI: 55-72) in non-EoE EGIDs. The percentage of persistent patients with non-EoE EGIDs was almost the same as those with EoE. Complications were more frequent in children than in adults.

Novel variants in the stem cell niche factor WNT2B define the disease phenotype as a congenital enteropathy with ocular dysgenesis

WNT2B is a member of the Wnt family, a group of signal transduction proteins involved in embryologic development and stem cell renewal and maintenance. We recently reported homozygous nonsense variants in WNT2B in three individuals with severe, neonatal-onset diarrhea, and intestinal failure. Here we present a fourth case, from a separate family, with neonatal diarrhea associated with novel compound heterozygous WNT2B variants. One of the two variants was a frameshift variant (c.423del [p.Phe141fs]), while the other was a missense change (c.722 G > A [p.G241D]) that we predict through homology modeling to be deleterious, disrupting post-translational acylation. This patient presented as a neonate with severe diet-induced (osmotic) diarrhea and growth failure resulting in dependence on parenteral nutrition. Her gastrointestinal histology revealed abnormal cellular architecture particularly in the stomach and colon, including oxyntic atrophy, abnormal distribution of enteroendocrine cells, and a paucity of colonic crypt glands. In addition to her gastrointestinal findings, she had bilateral corneal clouding and atypical genital development later identified as a testicular 46,XX difference/disorder of sexual development. Upon review of the previously reported cases, two others also had anterior segment ocular anomalies though none had atypical genital development. This growing case series suggests that variants in WNT2B are associated with an oculo-intestinal (and possibly gonadal) syndrome, due to the protein’s putative involvement in multiple developmental and stem cell maintenance pathways.

Glutamine Antagonist JHU-083 Normalizes Aberrant Hippocampal Glutaminase Activity and Improves Cognition in APOE4 Mice

Given the emergent aging population, the identification of effective treatments for Alzheimer's disease (AD) is critical. We investigated the therapeutic efficacy of JHU-083, a brain-penetrable glutamine antagonist, in treating AD using the humanized APOE4 knock-in mouse model. Cell culture studies were performed using BV2 cells and primary microglia isolated from hippocampi of adult APOE4 knock-in mice to evaluate the effect of JHU-083 treatment on LPS-induced glutaminase (GLS) activity and inflammatory markers. Six-month-old APOE4 knock-in mice were administered JHU-083 or vehicle via oral gavage 3x/week for 4-5 months and cognitive performance was assessed using the Barnes maze. Target engagement in the brain was confirmed using a radiolabeled GLS enzymatic activity assay, and electrophysiology, gastrointestinal histology, blood chemistry, and CBC analyses were conducted to evaluate the tolerability of JHU-083. JHU-083 inhibited the LPS-mediated increases in GLS activity, nitic oxide release, and pro-inflammatory cytokine production in cultured BV2 cells and primary microglia isolated from APOE4 knock-in AD mice. Chronic treatment with JHU-083 in APOE4 mice improved hippocampal-dependent Barnes maze performance. Consistent with the cell culture findings,postmortem analyses of APOE4 mice showed increased GLS activity in hippocampal CD11b+ enriched cells versus age-matched controls, which was completely normalized by JHU-083 treatment. JHU-083 was well-tolerated, showing no weight loss effect or overt behavioral changes. Peripheral nerve function, gastrointestinal histopathology, and CBC/clinical chemistry parameters were all unaffected by chronic JHU-083 treatment. These results suggest that the attenuation of upregulated hippocampal glutaminase by JHU-083 represents a new therapeutic strategy for AD.

Effect of the Use of Yarrowia lipolytica or Saccharomyces cerevisiae Yeast with a Probiotic in the Diet of Turkey Hens on Growth Performance and Gut Histology

Abstract The aim of the study was to determine whether the alternative yeast species Yarrowia lipolytica in turkey feed would have a more beneficial effect on growth performance and intestinal histology than the commonly used species Saccharomyces cerevisiae. An additional objective of the study was to test whether the addition of a probiotic to feed containing Yarrowia lipolytica or Saccharomyces cerevisiae yeast would enhance its effect on growth performance and intestinal histology in turkeys. The experiment was carried out on 480 turkey hens randomly divided into six groups. Birds from the control group (C) and group P were fed standard feeds but group P additionally received a probiotic (0.05%). Groups Y and YP received feed containing Yarrowia lipolytica fodder yeast (3%), and the YP group received also the probiotic (0.05%). Similarly, in groups S and SP, the turkeys received feed with Saccharomyces cerevisiae fodder yeast (3%), and for the SP group the probiotic was added to the feed (0.05%). Yarrowia lipolytica yeast added in the amount of 3% to the turkey feed may be an alternative to the commonly used Saccharomyces cerevisiae yeast, because it improved growth performance, and above all, had a more beneficial effect on intestinal histology. The use of Yarrowia lipolytica alone can be beneficial for growth performance, while the combined use of 3% Yarrowia lipolytica in the feed and a 0.05% addition of a probiotic containing Bacillus licheniformis and Bacillus subtilis has a more beneficial effect on gastrointestinal histology.

Postbiotic L. plantarum RG14 improves ruminal epithelium growth, immune status and upregulates the intestinal barrier function in post-weaning lambs

We investigate the effects of postbiotic Lactobacillus plantarum RG14 on gastrointestinal histology, haematology, mucosal IgA concentration, microbial population and mRNA expression related to intestinal mucosal immunity and barrier function. Twelve newly weaned lambs were randomly allocated to two treatment groups; the control group without postbiotic supplementation and postbiotic group with supplementation of 0.9% postbiotic in the diet over a 60-day trial. The improvement of rumen papillae height and width were observed in lambs fed with postbiotics. In contrast, no difference was shown in villi height of duodenum, jejunum and ileum between the two groups. Lambs received postbiotics had a lower concentration of IgA in jejunum but no difference in IgA concentration in serum and mucosal of the rumen, duodenum and ileum. In respect of haematology, postbiotics lowered leukocyte, lymphocyte, basophil, neutrophil and platelets, no significant differences in eosinophil. The increase in of IL-6 mRNA and decrease of IL-1β, IL-10, TNF mRNA were observed in the jejunum of lambs receiving postbiotics. Postbiotics also improved the integrity of the intestinal barrier by the upregulation of TJP-1, CLDN-1 and CLDN-4 mRNA. Postbiotic supplementation derived from L. plantarum RG14 in post-weaning lambs enhance the ruminal papillae growth, immune status and gastrointestinal health.

Microanatomy of the digestive tract, hooves and some visceral organs of addax antelope (Addax nasomaculatus) following a concentrate or forage feeding regime.

Subacute ruminal acidosis is a common disease in captive non-domesticated ruminants and is mainly diagnosed by rumen fluid pH and rumen histology. Furthermore, differences in ruminant gastrointestinal histology have been hypothesized to correlate with the browser-grazer continuum. Twelve surplus addax antelope (Addax nasomaculatus) were divided into two groups, fed either their usual diet, consisting of a concentrate feed with a limited amount of hay, or a diet of unlimited hay only, for 3months. After culling, descriptive and morphometric histology and pH measurements were compared between groups. Significant variations in cellular subpopulations were noted between groups, with roughage-fed individuals presenting primarily with balloon cells of the Stratum corneum and living layer cell vacuolization, whereas parakeratosis and intermediate-type cells were more frequent in the concentrate-fed group. Lesions typical of subacute ruminal acidosis were significantly more pronounced in concentrate-fed individuals. Ruminal pH measurements did not differ significantly, but were more consistent in forage-fed individuals, indicating a more stable reticuloruminal environment. The results indicate that ruminal histology may be more appropriate in assessing ruminal health compared to a single post-mortem pH measurement. It is proposed that Stratum corneum balloon cells may indicate cell maturation and not, as previously assumed, hyperfunction. Concentrate-fed individuals scored higher on the presence of inflammatory cells on hoof corium histology. The study further emphasizes the adaptability of ruminant digestive tract microanatomy in adult animals even after a short period of time and the positive effects an increased roughage diet may have in populations of captive grazing ruminants.

Role of comprehension on performance at higher levels of Bloom's taxonomy: Findings from assessments of healthcare professional students.

The first four levels of Bloom's taxonomy were used to create quiz questions designed to assess student learning of the gross anatomy, histology, and physiology of the gastrointestinal (GI) system. Information on GI histology and physiology was presented to separate samples of medical, dental, and podiatry students in computer based tutorials where the information from the two disciplines was presented either separately or in an integrated fashion. All students were taught GI gross anatomy prior to this study by course faculty as part of the required curriculum of their respective program. Student responses to the quiz questions were analyzed to assess both the validity of Bloom's cumulative hierarchy and the effectiveness of an integrated curriculum. No statistically significant differences were found between quiz scores from students who received the integrated tutorial and from those who received the separate tutorials. Multiple regression analyses provided partial support for a cumulative hierarchy where scores on the lower levels of Bloom's taxonomy predicted scores on higher levels. Notably, in the majority of regression analyses, the comprehension score was the key foundational predictor for application and analysis scores. This study supports the suggestion that educators increase the number of comprehension level questions, even at the expense of knowledge level questions, in course assessments both to evaluate lower order cognitive skills and also as a predictor of success on questions requiring application and analysis levels of the higher order cognitive skills of Bloom's taxonomy. Anat Sci Educ 11: 433-444. © 2018 American Association of Anatomists.

The Proof Is in the (Liver) Parenchyma: Primary Hepatic Presentation of Multiple Myeloma and Amyloidosis

Multiple myeloma (MM) and amyloidosis have known gastrointestinal (GI) involvement. Both present with protean features, making diagnosis a challenge. GI histology can confirm clandestine diagnoses. We present a patient whose liver biopsy led to diagnosis of systemic amyloidosis and multiple myeloma. It illustrates that primary GI involvement of infrequently encountered systemic illnesses can mimic common primary hepatic disorders. It challenges gastroenterologists to consider atypical systemic disorders when clinical courses do not respond as expected. A 49 year-old African American male with no liver disease presented with parasthesias, pruritis and 15-lb weight loss. Serology revealed alkaline phosphatase 434 IU/L AST 208 IU/L ALT 444 IU/L, total bilirubin 1.4 mg/dL total protein 9.4 g/dL. Direct bilirubin albumin INR, ANA, AMA, and anti-smooth muscle antibody (ASMA) were normal. Abdominal ultrasound (US) had heterogeneous echotexture with no ductal dilation. Alkaline phosphatase rose to 600 IU/L, prompting liver biopsy. Pathology showed mixed neutrophilic and plasmacytic infiltrate, some areas devoid of bile ductules. Prednisone and azathioprine were started for suspicion of autoimmune hepatitis (AIH) and primary sclerosing cholangitis overlap. Given weak response, mycophenolate mofetil and ursodiol were added. During hematology work-up for concomitant paraproteinemia he had cardiogenic shock. In conjunction with hematologic work-up, suspicion for amyloidosis was raised prompting review of prior liver biopsy. Congo-Red stained positive for amyloidosis. Myocardial biopsy depicted AL amyloidosis confirming the diagnosis of multiple myeloma with amylodosis. Prior to starting chemotherapy he died of cardiac arrest. Amyloidosis is a systemic disorder of extracellular protein fibril deposition. Hepatic manifestations are nonspecific including hepatomegaly and heterogenous echotexture on US. Elevated alkaline phosphatase occurs frequently and histologically, amyloidosis involves portal vessels. Our patient had a liver biopsy whose further review revealed amyloidosis. Initial interpretation of lymphocytic infiltrate suggested an autoimmune process however the hepatic infiltrate was likely myeloma as bone marrow biopsy had 20% plasma cell population. Minimal response to AIH treatment and multisystem involvement prompted reevaluation of the patient's disease, with correct diagnosis. This case depicts primary hepatic presentation of systemic MM and amyloid.Figure 1

Effects of alternative plant-based feeds on hepatic and gastrointestinal histology and the gastrointestinal microbiome of sablefish (Anoplopoma fimbria)

Sablefish (Anoplopoma fimbria) is a highly desired seafood product, which encourages development of sustainable aquaculture methods for this marine fish. Conventional marine fish-based feeds provide essential nutrients including long chain fatty acids for piscivorous species such as sablefish. Alternative terrestrial ingredients could reduce fishing pressures on pelagic species that are the source of fish meal and fish oil, and improve source sustainability. Using juvenile sablefish, we compared the effects of a standard fish-based diet to two diets that contained primarily terrestrial plant ingredients with flaxseed or corn oil replacing the added fish oil. After an 8-week trial feeding period, there were striking differences attributable to diet. Fish receiving the alternative feeds had lower weight gain and shorter length than fish receiving the fish-based feed, suggesting sablefish obtained lower nutrients from the alternative feeds. Among the histological differences, the intestinal mucosa was significantly less vacuolated and the frequency of intestinal mucous cells was reduced in alternative feed fish. The most dramatic lesions were observed in the liver, where severe bile duct hyperplasia (53%, flaxseed oil diet; 33%, corn oil diet), and hepatocellular lesions (nuclear pleomorphism/megalocytosis, regeneration, hypertrophy, clear cell foci) occurred in only alternative feed fish. The hepatic and biliary lesions indicate the alternative diets may be deficient or possibly harmful to sablefish. The bacterial community structures from corn oil fish showed much less diversity than those for the other diets, and the microbiome structures from the three diets were distinctly different from each other. The intestinal microbiome for the fish-based diet included the largest number of families (68), and these fish also had the largest number of unique bacterial families (11) compared to those for corn oil (two) or flaxseed oil (one) fish. Regardless of diet, the stomach and intestinal microbiomes differed significantly from each other, and the feed microbiome differed from all gastrointestinal communities, suggesting that feed is not a significant source of gut bacterial diversity. Similar to other teleosts, the sablefish gastrointestinal microbiome is dominated by Bacteroidetes, Firmicutes, and Proteobacteria. These results suggest that diet-induced shifts in microbiome can occur relatively quickly in sablefish, but the shifts may not be sufficiently adaptive or cannot overcome nutrient deficiencies. This multidisciplinary study demonstrates the utility of histology and microbiology in characterizing dietary effects for novel aquaculture species. Statement of relevancePhysiological assessments such as gastrointestinal microbiomes and histopathology can provide a near-term evaluation of the nutritional adequacy of alternative feeds, such as plant-based lipid diets for marine carnivorous finfish. This is the first examination of the microbiome of sablefish, a marine species under development for sustainable aquaculture.

Abstract 4957: Next generation sequencing demonstrates association between tumor suppressor gene aberrations and poor outcome in patients with cancer

Abstract Next generation sequencing is advancing at a breathtaking pace, and is transforming patient care by allowing physicians to customize and match treatment to their patients’ tumor alterations. Our goal was to study the association between key molecular alterations and outcome parameters. We retrospectively reviewed the clinicopathologic and clinical outcomes of 392 patients for whom molecular testing had been performed. Potential associations between clinical characteristics and outcome parameters (overall survival (OS), time to metastasis/recurrence, and best progression-free survival (PFS)) were examined. The Kaplan-Meier method and Cox regression models were used for our analysis, and results were subjected to internal validation using a resampling method (bootstrap analysis). In a multivariable analysis (Cox regression model), the parameters that were statistically associated with a poorer overall survival were the presence of metastases at diagnosis (P = 0.014), gastrointestinal histology (P&amp;lt;0.0001), PTEN (P&amp;lt;0.0001), and CDKN2A alterations (P = 0.0001). The variables associated with a shorter time to metastases/recurrence were gastrointestinal histology (P = 0.004), APC (P = 0.008), PTEN (P = 0.026) and TP53 (P = 0.044) alterations. TP53 (P = 0.003) and PTEN (P = 0.034) alterations were independent predictors of a shorter best PFS. A personalized treatment approach (matching the molecular aberration with a cognate targeted drug) also correlated with a longer best PFS (P = 0.046). Our study demonstrated that, across diverse cancers, anomalies in specific tumor suppressor genes (PTEN, CDKN2A, APC, and/or TP53) were independently associated with a worse outcome, as reflected by time to metastases/recurrence, best PFS on treatment, and/or overall survival. These observations suggest that molecular diagnostic tests may provide important prognostic information in patients with cancer. Citation Format: Maria Schwaederle, Gregory A. Daniels, David E. Piccioni, Paul T. Fanta, Richard B. Schwab, Kelly A. Shimabukuro, Barbara A. Parker, Razelle Kurzrock. Next generation sequencing demonstrates association between tumor suppressor gene aberrations and poor outcome in patients with cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4957. doi:10.1158/1538-7445.AM2015-4957