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Cefotaxime (CTX) is an easily detectable antibiotic pollutant in the water environment, but little is known about its toxic effects on aquatic invertebrates, especially on the intestine. Here, we determined the oxidative stress conditions of A. sinica under CTX exposure with five concentrations (0, 0.001, 0.01, 0.1 and 1 mg/L) for 14 days. After that, we focused on changes in intestinal tissue morphology and gut microbiota in A. sinica caused by CTX exposure at 0.01 mg/L. We found malondialdehyde (MDA) was elevated in CTX treatment groups, suggesting the obvious antibiotic-induced oxidative stress. We also found CTX exposure at 0.01 mg/L decreased the villus height and muscularis thickness in gut tissue. The 16S rRNA gene analysis indicated that CTX exposure reshaped the gut microbiota diversity and community composition. Proteobacteria, Actinobacteriota and Bacteroidota were the most widely represented phyla in A. sinica gut. The exposure to CTX led to the absence of Verrucomicrobia in dominant phyla and an increase in Bacteroidota abundance. At the genus level, eleven genera with an abundance greater than 0.1% exhibited statistically significant differences among groups. Furthermore, changes in gut microbiota composition were accompanied by modifications in gut microbiota functions, with an up-regulation in amino acid and drug metabolism functions and a down-regulation in xenobiotic biodegradation and lipid metabolism-related functions under CTX exposure. Overall, our study enhances our understanding of the intestinal damage and microbiota disorder caused by the cefotaxime pollutant in aquatic invertebrates, which would provide guidance for healthy aquaculture. [ABSTRACT FROM AUTHOR]

This study aimed to explore the impact of SCD Probiotics supplementation on biomolecule profiles and histopathology of ileum and colon tissues during a 30‐day intermittent fasting (IF) program. Male Sprague–Dawley rats, aged 24 months, underwent 18‐h daily fasting and received 3 mL (1 × 108 CFU) of SCD Probiotics. The differences in biomolecule profiles were determined using FTIR Spectroscopy and two machine learning techniques, Linear Discriminant Analysis (LDA) and Support Vector Machine (SVM), which showed significant differences with high accuracy rates. Spectrochemical bands indicating alterations in lipid, protein and nucleic acid profiles in both tissues. The most notable changes were observed in the group subjected to both IF and SCD Probiotics, particularly in the colon. Both interventions, individually and in combination, decreased protein carbonylation levels. SCD Probiotics exerted a more substantial impact on membrane dynamics than IF alone. Additionally, both IF and SCD Probiotics were found to have protective effects on intestinal structure and stability by reducing mast cell density and levels of TNF‐α and NF‐κB expression in ileum and colon tissues, thus potentially mitigating age‐related intestinal damage and inflammation. Furthermore, our results illustrated that while IF and SCD Probiotics individually instigate unique changes in ileum and colon tissues, their combined application yielded more substantial benefits. This study provides evidence for the synergistic potential of IF and SCD Probiotics in combating age‐related intestinal alterations. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Agro-Environment Science is the property of Journal of Agro-Environment Science Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

This study aims to investigate the effects of plasma exchange on the biomolecular profiles and histology of ileum and colon tissues in young and aged Sprague–Dawley male rats. Fourier transform infrared (FTIR) spectroscopy, linear discriminant analysis and support vector machine (SVM) techniques were employed to analyse the lipid, protein, and nucleic acid indices in young and aged rats. Following the application of young plasma, aged rats demonstrated biomolecular profiles similar to those of their younger counterparts. Histopathological and immunohistochemical assessments showed that young plasma had a protective effect on the intestinal tissues of aged rats, increasing cell density and reducing inflammation. Additionally, the expression levels of key inflammatory mediators tumour necrosis factor‐alpha and cyclooxygenase‐2 significantly decreased after young plasma administration. These findings underscore the therapeutic potential of young plasma for mitigating age‐related changes and inflammation in the intestinal tract. They highlight the critical role of plasma composition in the ageing process and suggest the need for further research to explore how different regions of the intestines respond to plasma exchange. Such understanding could facilitate the development of innovative therapies targeting the gastrointestinal system, enhancing overall health during ageing. [ABSTRACT FROM AUTHOR]

Cefotaxime (CTX) is an easily detectable antibiotic pollutant in the water environment, but little is known about its toxic effects on aquatic invertebrates, especially on the intestine. Here, we determined the oxidative stress conditions of A. sinica under CTX exposure with five concentrations (0, 0.001, 0.01, 0.1 and 1 mg/L) for 14 days. After that, we focused on changes in intestinal tissue morphology and gut microbiota in A. sinica caused by CTX exposure at 0.01 mg/L. We found malondialdehyde (MDA) was elevated in CTX treatment groups, suggesting the obvious antibiotic-induced oxidative stress. We also found CTX exposure at 0.01 mg/L decreased the villus height and muscularis thickness in gut tissue. The 16S rRNA gene analysis indicated that CTX exposure reshaped the gut microbiota diversity and community composition. Proteobacteria, Actinobacteriota and Bacteroidota were the most widely represented phyla in A. sinica gut. The exposure to CTX led to the absence of Verrucomicrobia in dominant phyla and an increase in Bacteroidota abundance. At the genus level, eleven genera with an abundance greater than 0.1% exhibited statistically significant differences among groups. Furthermore, changes in gut microbiota composition were accompanied by modifications in gut microbiota functions, with an up-regulation in amino acid and drug metabolism functions and a down-regulation in xenobiotic biodegradation and lipid metabolism-related functions under CTX exposure. Overall, our study enhances our understanding of the intestinal damage and microbiota disorder caused by the cefotaxime pollutant in aquatic invertebrates, which would provide guidance for healthy aquaculture.

Abstract Background Mycobacterium avium subsp. paratuberculosis infected animals show a variety of granulomatous lesions, from focal forms with well-demarcated granulomas restricted to the gut-associated lymphoid tissue (GALT), that are seen in the initial phases or latency stages, to a diffuse granulomatous enteritis, with abundant (multibacillary) or scant (paucibacillary) bacteria, seen in clinical stages. Factors that determine the response to the infection, responsible for the occurrence of the different types of lesion, are still not fully determined. It has been seen that regulatory T cells (Treg) play an important role in various diseases where they act on the limitation of the immunopathology associated with the immune response. In the case of paratuberculosis (PTB) the role of Treg lymphocytes in the immunity against Map is far away to be completely understood; therefore, several studies addressing this subject have appeared recently. The aim of this work was to assess, by immunohistochemical methods, the presence of Foxp3+ T lymphocytes in intestinal samples with different types of lesions seen in cows with PTB. Methods Intestinal samples of twenty cows showing the different pathological forms of PTB were evaluated: uninfected controls (n = 5), focal lesions (n = 5), diffuse paucibacillary (n = 5) and diffuse multibacillary (n = 5) forms. Foxp3+ lymphocyte distribution was assessed by differential cell count in intestinal lamina propria (LP), gut-associated lymphoid tissue (GALT) and mesenteric lymph node (MLN). Results A significant increase in the number of Foxp3+ T cells was observed in infected animals with respect to control group, regardless of the type of lesion. However, when the different categories of lesion were analyzed independently, all individuals with PTB lesions showed an increase in the amount of Foxp3+ T lymphocytes compared to the control group but this increase was only significant in cows with focal lesions and, to a lesser extent, in animals with diffuse paucibacillary forms. The former showed the highest numbers, significantly different from those found in cows with diffuse lesions, where no differences were noted between the two forms. No specific distribution pattern was observed within the granulomatous lesions in any of the groups. Conclusions The increase of Foxp3+ T cells in focal forms, that have been associated with latency or resistance to infection, suggest an anti-inflammatory action of these cells at these stages, helping to prevent exacerbation of the inflammatory response, as occurs in diffuse forms, responsible for the appearance of clinical signs.

Increasingly evidence seems that from the biological samples—mainly the summation of DSC scans of the human blood plasma—can be separated into main and well-known components by deconvolution method. This fact alone could be an important advance in thermoanalytical research, but mostly, it has not been detected in soft tissues yet. The other main reason was that in our previous studies, the histological examinations did not show any significant abnormalities in the intestinal wall muscle layer. Thus, the aim of current research was to measure the small intestinal muscular tissue scans by deconvolution method following different long warm and cold ischaemia animal experiments. Retrospectively, DSC curves obtained from the thermoanalysis of intestinal tissue in animal experiments investigated by deconvolution mathematical methods. Different warm ischaemic insults caused mild decrease after 3 and 6 h in Tm3 and Tm4 transitions in the myosin assigned transition and actin filament transition. After cold storage, the separated 5 melting components appeared in similar order, but the decreases were lesser than after warm ischaemia. Meanwhile, the calorimetric enthalpy which is a good monitor of intervention is decreased in a time-dependent manner after 1 and 3 h in warm and cold ischaemia cases. In contrast, the enthalpy increased above the control value after 6-h warm or cold tissue damage. These results confirmed that both warm and cold ischaemic injuries are detectable by deconvolution of DSC curves in the muscular intestinal layers. [ABSTRACT FROM AUTHOR]

Abstract Micro- and nanoplastics (MNP) can be found virtually everywhere around us in the biosphere and food chain, therefore humans are continuously exposed to MNP, mainly via inhalation and ingestion. Here, we have applied physiologically relevant human-based advanced in vitro models representing the lung (MucilAir™) and gut (InTESTine™ and Intestinal Explant Barrier Chip (IEBC)) to study membrane passage of various MNP and their potency to induce cytotoxic effects, barrier disturbances or pro-inflammatory cell activation. Selected MNP of various materials (polystyrene, polyethylene, nylon, car tire, and marine MNP collected from the ocean), shapes (spheres, fragments and fibers), sizes (0.05–100 μm), some of which were fluorescently labelled for tracking, were included. Without affecting cell viability, nylon fibers and (cleaned) HDPE (high density polyethylene) disrupted the MucilAir™ epithelial barrier. Luminal exposure to polystyrene particles (1 and 10 μm) and pristine HDPE fragments significantly decreased human colon tissue functionality. Furthermore, all polystyrene particles (0.05, 1 and 10 μm) affected tissue viability in porcine jejunum, ileum and colon tissue after 5 h exposure, and this was further confirmed in the IEBC after 24 h of exposure to 10 μm polystyrene particles and nylon fibers. Exposure to nylon fibers and its supernatant led to pro-inflammatory cell activation, as shown by increased IL-6 release in MucilAir™ and in human colon tissue after 96 or 24 h, respectively. Regarding transepithelial penetration of the MNP, permeability of 0.05 μm polystyrene spheres in the MucilAir™ lung cell model reached 3.6 ± 1.2% after 24 h. With 3.37 ± 0.46% after 5 h under static conditions and 5.5 ± 1.3% after 24 h under microfluidic conditions MNP permeability across intestinal tissue was highest for the largest (10 μm) polystyrene spheres. Confocal microscopy confirmed the translocation of MNP across the lung and intestinal epithelial barrier. In conclusion, we present a study revealing the passage of MNP over the epithelium of advanced in vitro models for the lung and intestine barrier. Furthermore, pro-inflammatory cell activation and disrupted barrier integrity were observed after exposure to several of the tested MNP. Future research is needed to further identify the effects of shape, size and material on these processes and subsequently the health effects of humans.

Background: Mycobacterium avium subsp. paratuberculosis infected animals show a variety of granulomatous lesions, from focal forms with well-demarcated granulomas restricted to the gut-associated lymphoid tissue (GALT), that are seen in the initial phases or latency stages, to a diffuse granulomatous enteritis, with abundant (multibacillary) or scant (paucibacillary) bacteria, seen in clinical stages. Factors that determine the response to the infection, responsible for the occurrence of the different types of lesion, are still not fully determined. It has been seen that regulatory T cells (Treg) play an important role in various diseases where they act on the limitation of the immunopathology associated with the immune response. In the case of paratuberculosis (PTB) the role of Treg lymphocytes in the immunity against Map is far away to be completely understood; therefore, several studies addressing this subject have appeared recently. The aim of this work was to assess, by immunohistochemical methods, the presence of Foxp3+ T lymphocytes in intestinal samples with different types of lesions seen in cows with PTB. Methods: Intestinal samples of twenty cows showing the different pathological forms of PTB were evaluated: uninfected controls (n = 5), focal lesions (n = 5), diffuse paucibacillary (n = 5) and diffuse multibacillary (n = 5) forms. Foxp3+ lymphocyte distribution was assessed by differential cell count in intestinal lamina propria (LP), gut-associated lymphoid tissue (GALT) and mesenteric lymph node (MLN). Results: A significant increase in the number of Foxp3+ T cells was observed in infected animals with respect to control group, regardless of the type of lesion. However, when the different categories of lesion were analyzed independently, all individuals with PTB lesions showed an increase in the amount of Foxp3+ T lymphocytes compared to the control group but this increase was only significant in cows with focal lesions and, to a lesser extent, in animals with diffuse paucibacillary forms. The former showed the highest numbers, significantly different from those found in cows with diffuse lesions, where no differences were noted between the two forms. No specific distribution pattern was observed within the granulomatous lesions in any of the groups. Conclusions: The increase of Foxp3+ T cells in focal forms, that have been associated with latency or resistance to infection, suggest an anti-inflammatory action of these cells at these stages, helping to prevent exacerbation of the inflammatory response, as occurs in diffuse forms, responsible for the appearance of clinical signs. [ABSTRACT FROM AUTHOR]

In intensive aquaculture systems, dissolved oxygen (DO) is the most critical and limiting factor for the development and health of aquatic organisms. Hypoxia is a common phenomenon in aquaculture. Its effects on fish’s growth and immune system have been widely reported, but the molecular mechanisms of hypoxic stress on damage to the fish gut are not fully understood. Additionally, studies on the effects of hypoxic stress on the intestinal metabolism of cobia (Rachycentron canadum) are still limited. Therefore, this study used cobia to explore the effects of hypoxic stress on intestinal metabolism. In this study, the combination of high-throughput RNA sequencing (RNA-Seq) and non-targeted liquid chromatography-mass spectrometry (LC-MS) based metabolomics analyses were used to investigate changes in key genes and metabolites in the intestinal tract of juvenile cobia after 28 days of hypoxic stress (DO: 3.15 ± 0.21 mg/L). Transcriptomics analysis revealed the significant enrichment of 62 metabolic pathways, including glutathione metabolism, fat digestion and absorption, bile secretion, glycerolipid metabolism, tricarboxylic acid cycle, glycolysis/gluconeogenesis, and other pathways. Metabolomics analysis revealed that most of the significantly differential metabolites were related to the metabolism of amino acids and lipids. The combined transcriptomics and metabolomics analyses showed that the sustained hypoxic stress could cause some degree of oxidative damage to the gut of cobia, thereby interfering with its digestive absorption and physiological and metabolic processes, such as abnormalities in antioxidant and detoxification functions, disturbances in the metabolisms of amino acids, lipids, and carbohydrates, and reduction of ion transport capacity. These findings might help understand the molecular mechanisms of the intestinal tract under hypoxic stress in cobia, thereby reducing the damage caused by hypoxic stress to farmed fish.

Extracellular vesicles (EVs) are small membranous particles that contribute to intercellular communications. Separating EVs from tissue is still a technical challenge. Here, we present a rigorous method for extracting EVs from intestinal tissue in a mouse intestinal ischemia/reperfusion (I/R) model, and for analyzing their miRNA content. The isolated EVs show a typical cup shape with a size peak of 120–130 nm in diameter, confirmed by TEM and NTA. They also express EV markers such as CD9, CD63, CD81, Tsg101 and Alix. Real-time qPCR confirmed that these pellets contain miRNAs related to I/R injury. Our study presents a practical way to isolate EVs from intestinal tissue which is suitable for downstream applications such as miRNA analysis, and provides a novel method for investigating the mechanism of intestinal I/R injury.

Copyright of Parasite (1252607X) is the property of EDP Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The article presents the results of evaluating commercial methods for extracting nucleic acids from pig intestinal tissues for the diagnosis of PED. The study was based on samples of small intestine tissues and faeces from 3–5 day old pigs which died from PED. Nucleic acid extraction was performed using commercial kits with different nucleic acid separation strategies based on: silicon-sorbent; silicate membrane fixed in a microcentrifuge column and magnetic balls. The studies were conducted in two stages. The first was a comparison of the results of the amplification of the obtained nucleic acid extracts from the homogenate of the intestines of piglets by using the above-mentioned commercial kits for the extraction of nucleic acids. For this purpose, samples of homogenate were used which in weight corresponded to the guideline for the application of the test kits. The second step was directed to determining the efficiency of extraction of DNA and RNA from homogenate samples with a weight of 10, 50, 100 and 200 mg. Determination of the optimal methodological strategy of nucleic acid extraction for the diagnosis of porcine epidemic diarrhea by PCR has been investigated. The results of the PCR studies of RNA of the PED virus and a unique pig DNA fragment indicate that the extraction of nucleic acids by commercial kits has different levels of efficiency and depends on different factors. According to the research, it was found that the most important of them are the adsorption capacity of the solid-phase sorbent, its configuration and nature, which binds RNA and DNA molecules, the type of sample from which extraction takes place, its volume, or the tissue mass used for extraction. Based on the obtained results, it has been found that the most effective PED virus RNA extraction is by “ArtBioTech”, “Bio Extract Column”, and “Viral DNA/RNA Extraction Kit”, and pig genomic DNA extraction by the “ArtBioTech” and “Viral DNA / RNA extraction Kit”.

The present investigation includes the study of protein concentration in intestinal tissue of coccidiosis infected broiler chicks orally inoculated by the infection of Eimeria tenella sporulated oocysts and treated with Mercurius corrosivus by using the methods of histochemical techniques. The treatment of homoeopathic medicine Mercurius corrosivus was administered to the coccidiosis infected group of broiler chicks.For the study of protein concentration in intestinal tissue of the coccidiosis infected and non-infected broiler chicks, the histochemical technique was used. Histochemical study indicates that the presence of different concentration of protein content found in the intestinal tissue of the broiler chicks.Intestinal tissue of all the chicks of different groups have presence of protein content in variable concentrations. It is also observed that the presence of protein concentration was variable in different region and shows different traces like minimum, medium and maximum amount of protein content found in different region of the intestinal tissue.The amount of protein concentration was observed high in group B(INC) in comparison to the other group of chicks observed during the experimental period. The concluded results of the study by using histochemical techniques shows that the protein concentration in the intestinal tissue is demonstrated by the variable staining reaction. The concentration of protein content is maximum in the parasitic stages showing the utilization of the protein from the host tissue. Whereas the infected and treated group with homoeopathic medicine Mercurius corrosivus also showed variable staining reaction according to the presence of protein in the intestinal tissue.

In this study, an in vivo animal experiment was performed, to investigate the intestinal absorption and distribution of naringin, hesperidin, and their metabolites by using UPLC-ESI-MS/MS and HPLC-PDA analysis. The bioavailability of naringin or hesperidin in duodenum, jejunum, ileum, cecum, and colon was calculated. A total of 7 and 13 metabolites were found and tentatively identified in intestinal tissue after orally administered naringin and hesperidin in mice, respectively. Sulfates of naringin could be absorbed by ileum and cecum, de-glycosylation metabolites could be absorbed by duodenum, and phenolic acids could be absorbed by the cecum and colon. Sulfates of hesperidin could be absorbed by duodenum and ileum, de-glycosylation metabolites could be absorbed by cecum, and phenolic acids could be absorbed by ileum, cecum and colon. Jejunum did not absorb metabolites of naringin or hesperidin. Results could be beneficial for understanding the intestinal bioactivities of naringin and hesperidin.

The gastrointestinal (GI) tract is a complex system responsible for nutrient absorption, digestion, secretion, and elimination of waste products that also hosts immune surveillance, the intestinal microbiome, and interfaces with the nervous system. Traditional in vitro systems cannot harness the architectural and functional complexity of the GI tract. Recent advances in organoid engineering, microfluidic organs-on-a-chip technology, and microfabrication allows us to create better in vitro models of human organs/tissues. These micro-physiological systems could integrate the numerous cell types involved in GI development and physiology, including intestinal epithelium, endothelium (vascular), nerve cells, immune cells, and their interplay/cooperativity with the microbiome. In this review, we report recent progress in developing micro-physiological models of the GI systems. We also discuss how these models could be used to study normal intestinal physiology such as nutrient absorption, digestion, and secretion as well as GI infection, inflammation, cancer, and metabolism.

The aim of this study was to evaluate the effects of biofloc (with addition of spoilage date palm extract (SDE) and molasses as sugar sources) on bacterial load of water, growth performance and intestine of Litopenaeus vannamei using diets containing different protein levels. Four biofloc treatments were designated including the biofloc treatment with SDE + a diet containing 25% protein (P25), biofloc treatment with molasses + a diet containing 25% protein (M25), biofloc treatment with SDE + a diet containing 15% protein (P15), biofloc treatment with molasses + a diet containing 15% protein (M15) and the control treatment with a diet containing 38% protein (Control). All treatments were conducted in triplicates. A 35-day study was conducted on 35 juvenile shrimp (with average weight of 5.37±0.33 g) which randomly stocked in fifteen tanks (each 300 L) at a stocking density of 175 shrimp m−3. The highest amounts of growth parameters (weight gain, Feed Conversion Ratio (FCR), growth rate and survival) were observed in P25 treatment and the lowest amounts of growth parameters were observed in the control (P

Immunofluorescence microscopy is an essential tool for tissue‐based research, yet data reporting is almost always qualitative. Quantification of images, at the per‐cell level, enables "flow cytometry‐type" analyses with intact locational data but achieving this is complex. Gastrointestinal tissue, for example, is highly diverse: from mixed‐cell epithelial layers through to discrete lymphoid patches. Moreover, different species (e.g., rat, mouse, and humans) and tissue preparations (paraffin/frozen) are all commonly studied. Here, using field‐relevant examples, we develop open, user‐friendly methodology that can encompass these variables to provide quantitative tissue microscopy for the field. Antibody‐independent cell labeling approaches, compatible across preparation types and species, were optimized. Per‐cell data were extracted from routine confocal micrographs, with semantic machine learning employed to tackle densely packed lymphoid tissues. Data analysis was achieved by flow cytometry‐type analyses alongside visualization and statistical definition of cell locations, interactions and established microenvironments. First, quantification of Escherichia coli passage into human small bowel tissue, following Ussing chamber incubations exemplified objective quantification of rare events in the context of lumen‐tissue crosstalk. Second, in rat jejenum, precise histological context revealed distinct populations of intraepithelial lymphocytes between and directly below enterocytes enabling quantification in context of total epithelial cell numbers. Finally, mouse mononuclear phagocyte—T cell interactions, cell expression and significant spatial cell congregations were mapped to shed light on cell–cell communication in lymphoid Peyer's patch. Accessible, quantitative tissue microscopy provides a new window‐of‐insight to diverse questions in gastroenterology. It can also help combat some of the data reproducibility crisis associated with antibody technologies and over‐reliance on qualitative microscopy. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC. on behalf of International Society for Advancement of Cytometry. [ABSTRACT FROM AUTHOR]

Intestinal mucosal immunity plays a vital role against Vibrio mimicus infection because it is an enteric pathogen causing serious vibriosis in fish. In the previous studies, we developed an oral double-targeted DNA vaccine of V. mimicus and demonstrated that the vaccine could elicit significantly higher intestinal mucosal immune response than did naked DNA vaccine. But, little is known underlying regulatory molecular mechanisms of the enhanced intestinal mucosal immunity. Here the transcriptome and proteome in the intestines of the grass carps immunized or not with the double-targeted DNA vaccine were investigated by using RNA-seq and iTRAQ-coupled LC-MS/MS. Compared with the control group, a total of 5339 differentially expressed genes (DEGs) and 1173 differentially expressed proteins (DEPs) were identified in the immunized fish intestines. Subsequently, the integrated analysis between transcriptome and proteome data revealed that 250 DEPs were matched with the corresponding DEGs (named associated DEPs/DEGs) at both transcriptome and proteome levels. Fifty of all the associated DEPs/DEGs were immune-related and mainly enriched in phagosome, antigen-processing and presentation, complement and coagulation cascades, NLRs and MAPK signaling pathways via Gene Ontology and KEGG pathway analyses, which suggested the coordination of the five activated pathways was essential to the enhanced intestinal mucosal immune response in the immunized fish. The protein-protein interaction analysis showed that 60 of the 63 immune-related DEPs to form an integrated network. Additionally, randomly selected DEGs and DEPs were respectively validated by quantitative real-time RT-PCR and multiple reaction monitoring (MRM) assay, indicating that the both RNA-Seq and iTRAQ results in the study were reliable. Overall, our comprehensive transcriptome and proteome data provide some key genes and their protein products for further research on the regulatory molecular mechanisms underlying the enhanced intestinal mucosal immunity. • First transcriptomic and proteomic analyses of grass carp intestine post-vaccination. • 5339 genes and 1173 proteins were significant differentially expressed. • 250 DEPs including 50 immune-related proteins were associated with their DEGs. • Phagosome, antigen processing-presentation, complement-coagulation cascades, NLRs and MAPK pathways were activated. • The data are useful for further study on the elevated intestinal mucosal immunity mechanism. [ABSTRACT FROM AUTHOR]

A carbonic anhydrase VII gene, encoding 277 amino acids, was identified in the intestinal tissue of pufferfish (Takifugu rubripes). The translated protein with an 833-bp complete coding sequence derived from the 1378-bp cloned sequence showed 83% identity with swamp eel CA VII, 76% with zebrafish CA VII, and 77% with coho salmon CA VII-like protein. The cloned protein also showed 68–69% identity with mammalian CA VII. The predicted molecular weight and iso-electric point of the protein were 30.84 kDa and 6.07, respectively. Active site analysis of the pufferfish CA VII indicated that most of the important residues involved in catalytic activity were highly conserved, whereas four cysteine residues at positions 55, 103, 184, and 275 differed from those in human CA II, and were related to cell-defense mechanisms against oxidative damage. Phylogenetic analysis showed that the cloned sequence was clustered within the fish CA VII clade and close to the swamp eel CA VII. Structural modeling of the pufferfish CA VII protein revealed the conservation of zinc binding histidine residues (zinc ion and histidine residues). Differential expression patterns of the pufferfish CA VII were determined with semiquantitative reverse transcription (RT)-PCR and quantitative PCR (q-PCR) as well. The results of q-PCR revealed that the pufferfish CA VII was highly expressed in intestinal tissue. The pufferfish CA VII was also detected within intestinal tissue sections using an in situ hybridization assay. [ABSTRACT FROM AUTHOR]

Inflammatory bowel disease (IBD) develops through complex interplay of genetic, microbial, immune, and environmental factors. Trace elements alterations are commonly present in IBD and may have influence on IBD development. Heavy metal pollution is one of the major environmental issues nowadays and IBD incidence is rising in countries where industry starts to develop. Metals are implicated in processes that are connected to IBD pathogenesis. The aim of this study was to investigate toxic and trace element levels in pediatric population of IBD patients both in serum and intestinal mucosa. This prospective study enrolled children newly diagnosed with IBD in University children's hospital in Belgrade. Concentrations of thirteen elements: Al, As, Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Se and Zn in serum and intestinal mucosa of 17 newly diagnosed children with IBD (10 Crohn's disease (CD) and 7ulcerative colitis (UC)) and 10 controls were assessed using inductively coupled plasma mass spectrometry (ICP-MS). Intestinal mucosa samples were taken from terminal ileum and six different colon segments (cecum, ascending colon, colon transversum, descending and sigmoid colon and rectum). The results demonstrated significant alterations in serum and intestinal mucosa concentrations of investigated elements. Serum iron was significantly decreased in IBD and CD group, compared to controls while serum Cu significantly differed between three investigated groups with highest concentration observed in CD children. Serum manganese was the highest in the UC subgroup. Terminal ileums of IBD patients contained significantly lower amount of Cu, Mg, Mn and Zn with Mn being significantly decreased also in CD patients compared to control. IBD patients' caecum contained significantly less Mg and Cu while colon transversum tissue samples from IBD and Crohn's patients contained significantly more chromium than controls. Moreover, sigmoid colon of IBD patients were poorer in Mg than controls (p < 0.05). Colon Al, As and Cd were significantly reduced in IBD, and UC children compared to control. Correlations of investigated elements in CD and UC groups were different from controls. Biochemical and clinical parameters showed correlation with element concentrations in intestines. Sera of CD, UC and control children significantly differ in Fe, Cu and Mn levels. Serum manganese was the highest in the UC subgroup creating the most prominent and only significant difference between UC and CD subgroups. Terminal ileum of IBD patients contained significantly lower amount of majority of investigated essential trace elements and toxic elements were significantly reduced in colon of IBD and UC patients. Investigation of macro- and microelement alterations in children and adults has potential to further elucidate IBD pathogenesis. [ABSTRACT FROM AUTHOR]

We investigated the expression pattern of Dkk-3, a secreted Wnt pathway inhibitor, in mouse intestinal tissue and three-dimensional cultured Caco-2 spheroids. Dkk-3 was expressed at the bottom of crypts from the mouse small intestine. Human colon adenocarcinoma Caco-2 cells expressed Dkk-3 under a semi-confluent condition, but Dkk-3 expression was seen in only some of the Caco-2 cells when the cells were sparse. Caco-2 cells formed hollow spheroids in Matrigel, and the layer-forming cells in the hollow spheroids expressed Dkk-3. Our results demonstrated that Dkk-3 might affect intestinal cells when the fate of stem cells changes.

Dihydromyricetin, 3,3′,4′,5,5′,7-hexahydroxyflavanone, is a phytochemical occurring in high quantities in tree Hovenia dulcis. This flavanonol is effective in counteracting acute EtOH intoxication and in reducing excessive EtOH consumption. As dihydromyricetin is considered for a potential human use its interactions with biotransformation enzymes should be examined. In general, ingested foreign compounds (xenobiotics) might stimulate expression (induction) of these enzymes and/or inhibit their activities. Usually the metabolism of xenobiotics proceeds via two phases of sequential enzymatic conversion to facilitate their excretion from the body. As xenobiotics biotransformation enzymes of the phase II, sulfo- and N/ O-acetyltransferases (SULTs and NATs), are involved in the process of carcinogen activation, their induction/inhibition by dihydromyricetin should be examined. Dihydromyricetin was administered to experimental rats by gastric gavages in three consecutive doses (60 mg/kg body weight/day). The induction of sulfo- and N/ O-acetyltransferases was assessed based on the protein levels on Western blots and on their metabolic activity in cytosolic samples of liver, small intestines and colon. Moreover, dihydromyricetin the inhibition of sulfo- and N/ O-acetyltransferase mediated activities was examined with recombinant enzymes. Dihydromyricetin induced SULT1A1 activity in all tissues were studied with the exception of small intestines but had no effect on N/ O-acetyltransferases. While dihydromyricetin did not affect SULT1A1 specific activity, both NAT1 and NAT2 were effectively inhibited (IC <10 μmol dm). Graphical abstract: [ABSTRACT FROM AUTHOR]

Background and aimIt has been reported that intestinal ischemia–reperfusion (I/R) injury results from oxidative stress caused by increased reactive oxygen species. Dexpanthenol (Dxp) is an alcohol analogue with epitelization, anti-inflammatory, antioxidant, and increasing peristalsis activities. In the present study, the aim was to investigate protective and therapeutic effects of Dxp against intestinal I/R injury.Materials and methodsOverall, 40 rats were assigned into five groups including one control, one alone Dxp, and three I/R groups (40-min ischemia; followed by 2-h reperfusion). In two I/R groups, Dxp (500 mg/kg, i.m.) was given before or during ischemia. The histopathological findings including apoptotic changes, and also tissue and serum biochemical parameters levels, were determined. Oxidative stress and ileum damage were assessed by biochemical and histological examination. In the control (n = 8) and alone Dxp (n = 8; 500 mg/kg, i.m. of Dxp was given at least 30 min before recording), groups were incised via laparotomy, and electrical activity was recorded from their intestines. In this experiment, the effect of Dxp on the motility of the intestine was examined by analyzing electrical activity.ResultsIn ileum, oxidant levels were found to be higher, while antioxidant levels were found to be lower in I/R groups when compared with controls. Dxp approximated high levels of oxidants than those in the control group, while it increased antioxidant values compared with I/R groups. Histopathological changes caused by intestinal I/R injury and histological improvements were observed in both groups given Dxp. In the Dxp group, electrical signal activity markedly increased compared with the control group.ConclusionsHere, it was seen that Dxp had protective and therapeutic effects on intestinal I/R injury and gastrointestinal system peristaltism. [ABSTRACT FROM PUBLISHER]

Lin Z, Hegarty JP, Cappel JA, Yu W, Chen X, Faber P, Wang Y, Kelly AA, Poritz LS, Peterson BZ, Schreiber S, Fan J-B, Koltun WA. Identification of disease-associated DNA methylation in intestinal tissues from patients with inflammatory bowel disease. Overwhelming evidence supports the theory that inflammatory bowel disease (IBD) is caused by a complex interplay between genetic predispositions of multiple genes, combined with an abnormal interaction with environmental factors. It is becoming apparent that epigenetic factors can have a significant contribution in the pathogenesis of disease. Changes in the methylation state of IBD-associated genes could significantly alter levels of gene expression, potentially contributing to disease onset and progression. We have explored the role of DNA methylation in IBD pathogenesis. DNA methylation profiles (1505 CpG sites of 807 genes) of matched diseased ( n = 26) and non-diseased ( n = 26) intestinal tissues from 26 patients with IBD [Crohn's disease (CD) n = 9, ulcerative colitis (UC) n = 17] were profiled using the GoldenGate™ methylation assay. After an initial identification of a panel of 50 differentially methylated CpG sites from a training set (14 non-diseased and 14 diseased tissues) and subsequent validation with a testing set (12 non-diseased and 12 diseased tissues), we identified seven CpG sites that are differentially methylated in intestinal tissues of IBD patients. We have also identified changes in DNA methylation associated with the two major IBD subtypes, CD and UC. This study reports IBD-associated changes in DNA methylation in intestinal tissue, which may be disease subtype-specific. [ABSTRACT FROM AUTHOR]

This paper describes a means to predict the absorbability of poorly water-soluble drugs in humans based on rat intestinal permeability assessed by the in vitro Ussing-type chamber method. We investigated the correlation between the apparent permeability coefficients (Papp) of 10 water-soluble drugs obtained by the in vitro chamber method, in which the excised rat small intestinal tissue was used as the membrane, and the fractions absorbed (Fa) in humans. Using this correlation, we predicted Fa values of 5 poorly water-soluble drugs based on their Papp obtained through our modified chamber method using an additive. For water-soluble drugs, a good correlation between Papp and Fa, expressed by the equation: Fa=1-exp(-Papp×1.51×105)(r2=0.920), was found. The poorly water-soluble drugs used in the present study could be solubilized with 5% (final concentration) dimethylsulfoxide, and their Papp could be obtained through our modified chamber method. For poorly water-soluble drugs whose dose:solubility ratio ranged from 2500 to 3500 ml, predicted Fa values were favorably comparable with their Fa values reported in humans in the literature. These results showed that the in vitro Ussing-type chamber method was a useful method for predicting the Fa of poorly water-soluble drugs. [Copyright &y& Elsevier]

Objective: To study the expression and significance of follistatin-like protein 1(FSTL1) in peripheral blood and intestinal tissues of patients with ulcerative colitis(UC). Methods: Patients with ulcerative colitis in our hospital from March 2013 to March 2014 were selected as the study group(n=60), and subjects with normal pathology and colonoscopy were chosen as the control group(n=60). FSTL1 levels in the peripheral blood of the two groups were detected by ELISA, and the FSTL1 level in the intestinal tissues of the two groups were detected by the immunohistochemical staining. Then the correlation between the ulcerative colitis activity index(DAI) and FSTI1 were analyzed by Pearson correlation analysis. Results: The plasma FSTL1 level of the study group was significantly higher than that of the control group with statistically significant difference(t=11.063, P=0.025). The positive rate of FSTL1 in the intestinal tissues of the study group was significantly higher than that of the control group with statistically significant difference(x2=12.987, P=0.019); DAI and FSTI1 of the study group was positively correlated(r=0.719, P=0.023). Conclusion: The FSTL1 expression level was elevated in patients with ulcerative colitis, and was positively correlated with disease activity. [ABSTRACT FROM AUTHOR]

• Naringin and hesperidin, and their metabolites were found to be widely absorbed in the intestinal tissue. • Both naringin and hesperidin have different bioavailability in each intestinal tissue. • There were different metabolites distributed in different intestinal tissues. In this study, an in vivo animal experiment was performed, to investigate the intestinal absorption and distribution of naringin, hesperidin, and their metabolites by using UPLC-ESI-MS/MS and HPLC-PDA analysis. The bioavailability of naringin or hesperidin in duodenum, jejunum, ileum, cecum, and colon was calculated. A total of 7 and 13 metabolites were found and tentatively identified in intestinal tissue after orally administered naringin and hesperidin in mice, respectively. Sulfates of naringin could be absorbed by ileum and cecum, de-glycosylation metabolites could be absorbed by duodenum, and phenolic acids could be absorbed by the cecum and colon. Sulfates of hesperidin could be absorbed by duodenum and ileum, de-glycosylation metabolites could be absorbed by cecum, and phenolic acids could be absorbed by ileum, cecum and colon. Jejunum did not absorb metabolites of naringin or hesperidin. Results could be beneficial for understanding the intestinal bioactivities of naringin and hesperidin. [ABSTRACT FROM AUTHOR]

Small intestinal anastomoses are commonly performed in veterinary medicine following resection of diseased or devitalized intestinal tissue. Traditionally, suture has been employed to anastomose intestinal ends. However, use of intestinal staplers has become increasingly popular due to the ability to produce a rapid anastomosis with purported superior healing properties. Under normal conditions, intestinal healing occurs in three phases: inflammatory, proliferative, and maturation. Dehiscence, a devastating consequence of intestinal anastomosis surgery, most often occurs during the inflammatory phase of healing where the biomechanical strength of the anastomosis is almost entirely dependent on the anastomotic technique (suture or staple line). The resulting septic peritonitis is associated with a staggering morbidity rate upwards of 85% secondary to the severe systemic aberrations and financial burden induced by septic peritonitis and requirement of a second surgery, respectively. Intraoperative and postoperative consideration of the multifactorial nature of dehiscence is required for successful patient management to mitigate recurrence. Moreover, intensive postoperative critical care management is necessitated and includes antibiotic and fluid therapy, vasopressor or colloidal support, and monitoring of the patient's fluid balance and cardiovascular status. An understanding of anastomotic techniques and their relation to intestinal healing will facilitate intraoperative decision-making and may minimize the occurrence of postoperative dehiscence. [ABSTRACT FROM AUTHOR]

Small intestinal foreign body obstructions occur commonly in dogs, accounting for 80% of all canine intestinal obstructions. Such obstructions induce local aberrations in secretion, absorption, and intestinal motility that can precipitate devastating systemic consequences, including a systemic inflammatory response, sepsis, and multiorgan dysfunction. Radiographic diagnosis is poorly sensitive relative to ultrasonography for diagnosing the presence of obstructive foreign material. Emergent surgical intervention is indicated for dogs with obstructive foreign material due to an inability to assess the degree of compromise of the intestinal wall that may precipitate intestinal perforation and to mitigate progression of life-threatening electrolyte and acid-base imbalances secondary to sequestration and emesis. Intraoperatively, an enterotomy or resection and anastomosis may be required to remove the obstructive material. A number of subjective and objective techniques for assessing the viability of intestinal tissue have been described due to the poor accuracy associated with surgeon assessment of color, peristalsis, pulsation, bleeding, and mural thickness alone. Such techniques have the potential to alter the surgeon's decision-making regarding performance of an enterotomy or resection and anastomosis, potentially reducing morbidity associated with intestinal surgery. [ABSTRACT FROM AUTHOR]

The intestine is often examined histologically in connection with allergies and in search for pathological changes. To be able to examine the intestine histologically with a microscope, it must be sampled and processed correctly. For microscopic analysis, the samples have to be cut into thin sections, stained, and mounted on slides. Since it is not possible to cut fresh samples without damaging them, they must first be fixed. The most common method, which is described herein, is the fixation in formalin with subsequent embedding in paraffin and staining of the slides with hematoxylin and eosin (H&E). Hematoxylin solutions (in this case Mayer's hemalum solution) stain the acidic components of the cell, i.e., cell nuclei, blue. The staining with eosin gives a pink staining of cytoplasm. This chapter describes the method of processing intestinal tissue for paraffin-embedding, sectioning, and staining with H&E. Tissue processing can be done in tissue processing machines or manually. We describe the manual processing that is often used for smaller batches of samples.