Intestinal Sites Research Articles

Clinical Features and Prognosis of Patients with Primary Intestinal B-cell Lymphoma Treated with Chemotherapy with or without Surgery.

This multicenter study reviewed the clinical features and prognosis according to the primary site of involvement and the treatment modality in patients with B-cell primary intestinal lymphoma (PIL). Among 125 consecutive patients diagnosed with PIL, 100 patients were analyzed. The median age was 59 years, and the male to female ratio was 1.86:1. Diffuse large B-cell lymphoma (66/100, 66.0%) was the most common histological subtype. The estimated 5-year survival rate (5-YSR) was 48.5%. The 5-YSR was similar regardless of the type of primary treatment (chemotherapy alone vs. surgery/chemotherapy, 50.7 vs. 45.3%, p=0.582). A comparison of the survival according to the primary site of involvement revealed a 5-YSR of 32.5% (p=0.027), 64.3% (reference), 46.5% (p=0.113), and 49.8% (p=0.024) for the small intestine, ileocecal region, large intestine, and multiple sites, respectively. Multivariate analysis, however, revealed a low hemoglobin level, advanced Ann Arbor stage, and aggressive histological type to be independent prognostic factors for shorter survival but not ileocecal region involvement. The Ann Arbor stage, hemoglobin level, and histological type were independent prognostic factors for survival, while the primary site of involvement and treatment modality did not affect the prognosis in patients with B-cell PIL.

Understanding of the Site-Specific Microbial Patterns towards Accurate Identification for Patients with Diarrhea-Predominant Irritable Bowel Syndrome

ABSTRACTFecal microbial community could not fully represent the intestinal microbial community. However, most studies analyzing diarrhea-dominant irritable bowel syndrome (IBS-D) were mainly based on fecal samples. We aimed to characterize the IBS-D microbial community patterns using samples at multiple intestinal sites. This study recruited 74 IBS-D patients and 20 healthy controls (HC). 22.34%, 8.51%, 14.89%, and 54.26% of them contributed to one, two, three, and four sites: duodenal mucosa (DM), duodenal lumen (DL), rectal mucosa (RM), and rectal lumen (RL) of intestinal samples, respectively. Then 16S rRNA gene analysis was performed on these 283 samples. The result showed that IBS-D microbial communities have specific patterns at each intestinal site differing from that of HC. Across hosts and sites, Bacillus, Burkholderia, and Faecalibacterium were the representative genera in duodenum of IBS-D, duodenum of HC, and rectum of HC, respectively. Samples from mucosa and lumen in rectum were highly distinguishable, regardless of IBS-D and HC. Additionally, IBS-D patients have lower microbial co-abundance network connectivity. Moreover, RM site-specific biomarker: Bacteroides used alone or together with Prevotella and Oscillospira in RM showed outstanding performance in IBS-D diagnosis. Furthermore, Bacteroides and Prevotella in RM were strongly related to the severity of abdominal pain, abdominal discomfort, and bloating in IBS-D patients. In summary, this study also confirmed fecal microbial community could not fully characterize intestinal microbial communities. Among these site-specific microbial communities, RM microbial community would be more applicable in the diagnosis of IBS-D.IMPORTANCE Microbial community varied from one site to another along the gastrointestinal tract, but current studies about intestinal microbial community in IBS-D were mainly based on fecal samples. Based on 283 intestinal samples collected from DM, DL, RM, and RL of HC and IBS-D, we found different intestinal sites had their site-specific microbial patterns in IBS-D. Notably, RM site-specific microbes Bacteroides, Prevotella, and Oscillospira could be used to discriminate IBS-D from HC accurately. Our findings could help clinicians realize the great potential of the intestinal microbial community in RM for better diagnosis of IBS-D patients.

Regionally Distinct Immune and Metabolic Transcriptional Responses in the Bovine Small Intestine and Draining Lymph Nodes During a Subclinical Mycobacterium avium subsp. paratuberculosis Infection.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative infectious agent of Johne’s disease (JD), an incurable granulomatous enteritis affecting domestic livestock and other ruminants around the world. Chronic MAP infections usually begin in calves with MAP uptake by Peyer’s patches (PP) located in the jejunum (JE) and ileum (IL). Determining host responses at these intestinal sites can provide a more complete understanding of how MAP manipulates the local microenvironment to support its long-term survival. We selected naturally infected (MAPinf, n=4) and naive (MAPneg, n=3) cows and transcriptionally profiled the JE and IL regions of the small intestine and draining mesenteric lymph nodes (LN). Differentially expressed (DE) genes associated with MAP infection were identified in the IL (585), JE (218), jejunum lymph node (JELN) (205), and ileum lymph node (ILLN) (117). Three DE genes (CD14, LOC616364 and ENSBTAG00000027033) were common to all MAPinf versus MAPneg tissues. Functional enrichment analysis revealed immune/disease related biological processes gene ontology (GO) terms and pathways predominated in IL tissue, indicative of an activated immune response state. Enriched GO terms and pathways in JE revealed a distinct set of host responses from those detected in IL. Regional differences were also identified between the mesenteric LNs draining each intestinal site. More down-regulated genes (52%) and fewer immune/disease pathways (n=5) were found in the ILLN compared to a higher number of up-regulated DE genes (56%) and enriched immune/disease pathways (n=13) in the JELN. Immunohistochemical staining validated myeloid cell transcriptional changes with increased CD172-positive myeloid cells in IL and JE tissues and draining LNs of MAPinf versus MAPneg cows. Several genes, GO terms, and pathways related to metabolism were significantly DE in IL and JE, but to a lesser extent (comparatively fewer enriched metabolic GO terms and pathways) in JELN suggesting distinct regional metabolic changes in IL compared to JE and JELN in response to MAP infection. These unique tissue- and regional-specific differences provides novel insight into the dichotomy in host responses to MAP infection that occur throughout the small intestine and mesenteric LN of chronically MAP infected cows.

Directly recruited GATA6\u2009+\u2009peritoneal cavity macrophages contribute to the repair of intestinal serosal injury

Recruitment of bone marrow derived monocytes via bloodstream and their subsequent conversion to CX3CR1+ macrophages in response to intestinal injury is dependent on CCR2, Nr4a1, and the microbiome. This process is critical for proper tissue repair; however, GATA6+ peritoneal cavity macrophages might represent an alternative, more readily available source of mature and functional myeloid cells at the damaged intestinal locations. Here we show, using spinning-disk confocal microscopy, that large F4/80hiGATA6+ peritoneal cavity macrophages promptly accumulate at damaged intestinal sites upon intestinal thermal injury and upon dextran sodium sulfate induced colitis in mice via a direct route from the peritoneal cavity. In contrast to bloodstream derived monocytes/macrophages, cavity macrophages do not depend on CCR2, Nr4a1 or the microbiome for recruitment, but rather on the ATP-release and exposed hyaluronan at the site of injury. They participate in the removal of necrotic cells, revascularization and collagen deposition and thus resolution of tissue damage. In summary, peritoneal cavity macrophages represent a rapid alternative route of intestinal tissue repair to traditional monocyte-derived macrophages.

Intestinal metabolism and absorption mechanism of multi-components in Gaultheria leucocarpa var. yunnanensis - An assessment using in situ and in vitro models, comparing gut segments in pathological with physiological conditions

Ethnopharmacological relevanceDianbaizhu (Gaultheria leucocarpa var. yunnanensis) as a Chinese folk medicine exerts significant treatment effects on rheumatoid arthritis (RA) with a long historical time. Our previous reports showed that the anti-rheumatic arthritis fraction (ARF) extracted and enriched from Dianbaizhu possessed good druggability, which was better than its single active ingredients. However, the intestinal transport characteristics and mechanism of ARF have not been elucidated to date. Aim of the studyIn order to illustrate the role of active ingredients of ARF in alleviating RA and promoting the development of dosage forms, the intestinal metabolism, absorption properties and mechanism of ARF in vitro and in situ models were investigated. Materials and methodsFirstly, after incubating with 4 intestinal segments (duodenum, jejunum, ileum, and colon), 7 key components in ARF, including MATG-B, (+)-catechin, MSTG-A, Gaultherin, chlorogenic acid, quercetin, and kaempferol were quantitatively analyzed by a high-performance liquid chromatography (HPLC). Secondly, combining the physiological and pathological rats, the in situ single-pass intestinal perfusion and in vitro everted gut sacs of rats were performed to investigate the absorption features and transport mechanisms of ARF using HPLC and HPLC-Q-TOF-MS/MS. Subsequently, in situ studies were employed to determine the effect of P-glycoprotein (P-gp) inhibitor (verapamil) on the transport characteristics of ARF in RA model rats. ResultsComparing the absorption parameters of ARF incubated in different intestinal segments, data showed that the absorption of ARF in the small intestine was significantly stronger than that of the colon (P < 0.01). The number of characterized prototype components was subjected to the incubation time, drug concentration and rat body condition, but not the intestinal segments. There were no significant differences in the number of metabolites among different intestinal segments, administration concentrations and incubation time. The best small intestinal absorption site of ARF was duodenum and ileum in normal and model rats, respectively. The Peff values of 7 index compounds were all higher than 0.2 × 10−4cm/s, and the Fa values of 7 index compounds were all greater than 20% in the in situ perfusion investigation. The results showed that MSTG-B, MSTG-A and Gaultherin were likely to be substrates of P-gp as verapamil significantly enhanced their Peff and Ka values, while other ingredients were not P-gp substrates. ConclusionsThe intestinal membrane permeability of ARF was good. Its intestinal absorption mechanisms mainly involved active transportation processes and passive diffusion. Besides, this report provided data support and basis for clinical development, bioavailability improvement and formulation design.

Nanofiber could deliver lactic acid bacteria to the intestine of ruminant in vitro experiment.

This study investigates the use of nanofiber microcapsules produced by electrostatic spinning as a carrier for the delivery of lactic acid bacteria (LAB) to the intestine of ruminants. We hypothesized that the LAB encapsulated into nanofiber microcapsules can be delivered to a ruminant's intestinal tract with little effect on the rumen fermentation and related bacteria. The in vitro experiment included three treatments: control group; 0.01g Lactobacillus acidophilus NCFM (L. acidophilus NCFM) encapsulated in nanofiber microcapsules by electrostatic spinning group (ELAN, 2.0×1011 CFU/g); and 0.01g L. acidophilus NCFM powder group (LANP, 2.0×1011 CFU/g), each incubated with 30ml of buffer rumen fluid for 48h to determine the effect on rumen fermentation, then the abundance of L. acidophilus NCFM in the intestine was estimated using the modified in vitro three-step procedure. Treatment responses were statistically analysed using one-way ANOVA. The results showed that compared to the control, the ELAN group had a significant increase in pH (p<0.05), while the LANP group had a non-significant decrease in pH (p>0.05). LANP and ELAN groups had no significant influence on total volatile fatty acid and individual volatile fatty acids (p>0.05), apart from isobutyric acid of both groups, which reduced (p<0.05). ELAN group had a decreasing trend of gas production and dry matter digestion, while the LANP group increased them significantly (p<0.05). During the 16h and 48h rumen incubation, compared with control, there was no significant change in all bacteria in the ELAN group (p>0.05), while the LANP group increased the relative abundance levels of S. bovis, S. ruminantium, M. elsdenii, F. succinogenes, B. fibrisolvens, Lactobacillus, L. acidophilus NCFM (p<0.05). In the intestinal part, compared with control, the relative abundance of L. acidophilus NCFM in the ELAN group increased significantly (p<0.05), while the result was not observed in the LANP group. We concluded based on our findings that L. acidophilus NCFM could be protected by nanofiber microcapsules and delivered to the intestinal site with little influence on the rumen fermentation and bacterial community, suggesting nanofiber microcapsules prepared by electrospinning technology could be used as a carrier for rumen-protected study.

Expression of SARS-CoV-2 entry factors, electrolyte, and mineral transporters in different mouse intestinal epithelial cell types.

Angiotensin‐converting enzyme 2 (ACE2) and transmembrane proteases (TMPRSS) are multifunctional proteins required for SARS‐CoV‐2 infection or for amino acid (AA) transport, and are abundantly expressed in mammalian small intestine, but the identity of the intestinal cell type(s) and sites of expression are unclear. Here we determined expression of SARS‐CoV‐2 entry factors in different cell types and then compared it to that of representative AA, electrolyte, and mineral transporters. We tested the hypothesis that SARS‐CoV‐2, AA, electrolyte, and mineral transporters are expressed heterogeneously in different intestinal cell types by making mouse enteroids enriched in enterocytes (ENT), goblet (GOB), Paneth (PAN), or stem (ISC) cells. Interestingly, the expression of ACE2 was apical and modestly greater in ENT, the same pattern observed for its associated AA transporters B0AT1 and SIT1. TMPRSS2 and TMPRSS4 were more highly expressed in crypt‐residing ISC. Expression of electrolyte transporters was dramatically heterogeneous. DRA, NBCe1, and NHE3 were greatest in ENT, while those of CFTR and NKCC1 that play important roles in secretory diarrhea, were mainly expressed in ISC and PAN that also displayed immunohistochemically abundant basolateral NKCC1. Intestinal iron transporters were generally expressed higher in ENT and GOB, while calcium transporters were expressed mainly in PAN. Heterogeneous expression of its entry factors suggests that the ability of SARS‐CoV‐2 to infect the intestine may vary with cell type. Parallel cell‐type expression patterns of ACE2 with B0AT1 and SIT1 provides further evidence of ACE2's multifunctional properties and importance in AA absorption.

English

BACKGROUND Liver abscess has been recognised since the time of Hippocrates. Liver abscess is defined as collection of purulent material in liver parenchyma. They are usually caused by bacterial and amoebic infections, and less commonly, by other protozoal and helminthic organisms. Amoebic liver abscess is the commonest extra intestinal site of invasive amoebiasis which mainly affects infants and young children. The incidence of pyogenic liver abscess is much higher among children in developing countries than those in developed countries. The purpose of this study was to evaluate culture sensitivity pattern (Blood &amp; Pus) of liver abscess in children. METHODS This prospective observational study was conducted in the Department of Paediatrics, Chacha Nehru Bal Chikistalaya, Delhi from July 2016 –to August 2017. This study has got Institutional Ethics Committee approval (Regd No: IEC/MAMC/78, Dt: 26/07/2016). All children aged 1 month to 12 years admitted with liver abscess (included consecutively) were enrolled after considering inclusion and exclusion criteria. Written and informed consent was taken from parents/guardians of children. Their clinical characteristics, radiological features, laboratory data, clinical management, and outcomes were analysed. RESULTS In our study, out of 70 patients, 3.2 % patients showed growth in the blood culture. Organsims isolated were Methicillin resistant Staphylococcus aureus (MRSA) 1.4 % (1), Salmonella typhi 1.4 % (1), staphylococcus coagulase negative 1.4 % (1). Out of 70 patients of liver abscess enrolled in the study, 36 patients underwent aspiration of pus from the abscess. Out of 36 aspirated cases, gram positive cocci was identified in 1 (1.4 %) patient. In our study, no acid fast bacilli was identified and no fungal culture showed growth of organism. Out of 70 cases of liver abscess, 10 were found to be amoebic liver abscess. In our study, all the 70 patients were started on empirical antibiotics. Out of 70 patients, surgical intervention was done in 36 patients. In our study all the patients were started on empirical antibiotics according to hospital protocol. CONCLUSIONS Liver abscess should be considered in children presenting with fever and abdominal pain. Organisms recovered from liver abscesses vary greatly. Surgical drainage has been the traditional mode of treatment of pyogenic liver abscess, but this was replaced by IV broad-spectrum antibiotics and imaging-guided percutaneous drainage. KEYWORDS Paediatric Liver Abscess, Amoebic Liver Abscess, Pyogenic Liver Abscess, Culture-Sensitivity, Children

Synergy between "Probiotic" Carbon Quantum Dots and Ciprofloxacin in Eradicating Infectious Biofilms and Their Biosafety in Mice.

Orally administrated probiotic bacteria can aid antibiotic treatment of intestinal infections, but their arrival at their intestinal target site is hampered by killing in the gastrointestinal tract and by antibiotics solely intended for pathogen killing. Carbon-quantum-dots are extremely small nanoparticles and can be derived from different sources, including bacteria. Here, we hypothesize that carbon-quantum-dots inherit antibacterial activity from probiotic source bacteria to fulfill a similar role as live probiotics in intestinal infection therapy. Physico-chemical analyses indicated that carbon-quantum-dots, hydrothermally derived from Bifidobacterium breve (B-C-dots), inherited proteins and polysaccharides from their source-bacteria. B-C-dots disrupted biofilm matrices of Escherichia coli and Salmonella typhimurium biofilms through extensive reactive-oxygen-species (ROS)-generation, causing a decrease in volumetric bacterial-density in biofilms. Decreased bacterial densities leave more open space in biofilms and have enhanced ciprofloxacin penetration and killing potential in an E. coli biofilm pre-exposed to probiotic B-C-dots. Pathogenic carbon-quantum-dots hydrothermally derived from E. coli (E-C-dots) did not disrupt pathogenic biofilms nor enhance E. coli killing potential by ciprofloxacin. B-C-dots were biosafe in mice upon daily administration, while E-C-dots demonstrated a decrease in white blood cell and platelet counts and an increase in C-reactive protein levels. Therefore, the way is paved for employing probiotic carbon-quantum-dots instead of viable, probiotic bacteria for synergistic use with existing antibiotics in treating intestinal infections.

Effects of Deoxynivalenol and Fumonisins on Broiler Gut Cytoprotective Capacity.

Mycotoxins are a crucial problem for poultry production worldwide. Two of the most frequently found mycotoxins in feedstuffs are deoxynivalenol (DON) and fumonisins (FUM) which adversely affect gut health and poultry performance. The current knowledge on DON and FUM effects on broiler responses relevant for gut detoxification, antioxidant capacity, and health is still unclear. The aim of this study was to assess a range of selected molecular intestinal biomarkers for their responsiveness to the maximum allowable European Union dietary levels for DON (5 mg/kg) and FUM (20 mg/kg) in broilers. For the experimental purpose, a challenge diet was formulated, and biomarkers relevant for detoxification, antioxidant response, stress, inflammation, and integrity were profiled across the broiler intestine. The results reveal that DON significantly (p < 0.05) induced aryl hydrocarbon receptor (AhR) and cytochrome P450 enzyme (CYP) expression mainly at the duodenum. Moreover, DON and FUM had specific significant (p < 0.05) effects on the antioxidant response, stress, inflammation, and integrity depending on the intestinal segment. Consequently, broiler molecular responses to DON and FUM assessed via a powerful palette of biomarkers were shown to be mycotoxin and intestinal site specific. The study findings could be highly relevant for assessing various dietary bioactive components for protection against mycotoxins.

PSIII-21 Efficacy of deoxynivalenol detoxification by sodium metabisulfite (SMBS) containing nanofibrous mats using an in vitro intestinal epithelial cell IPEC-J2

Abstract Deoxynivalenol (DON) occurs in many commonly used cereal grains. Pigs feed with DON concentrations as low as 0.6–2.0 mg/kg can result in reduced feed intake and growth rate, damage to intestinal epithelial cells, and increase susceptibility to enteric pathogens. Sodium metabisulfite (SMBS) can efficiently detoxify DON by converting it 8-DONS or 10-DONS in vitro. However, if SMBS is added directly to the feed, SMBS rapidly degrades under acidic aqueous conditions (e.g. pig stomach) and little SMBS is delivered to the intestinal absorption site where it can effectively detoxify DON. Thus, the objective of this study was to encapsulate SMBS into Eudragit L100-55 nanofibrous mats to deliver intact SMBS to the small intestine and evaluate the efficacy of DON detoxification in the simulated intestine fluid (SIF) using an in vitro intestinal epithelial cell (IPEC-J2) model. Nanofibrous mats were produced by coaxial electrospinning, with peak loading capacity and loading efficiency of SMBS reaching 32.00% and 80.01%. DON-induced cytotoxicity was not observed during in vitro analysis consisting of incubation of DON in the presence of SMBS-containing nanofibers (0.5% w/w) in simulated gastric fluid (SGF) for 2 h followed by incubation in a mixture of SGF and SIF (1:1) for 20 min. Meanwhile, compared to the DON treatment, incubation of DON in the presence of SMBS-containing nanofiber (0.5% w/w) in SGF for 2 h and SIF for 20 min decreased the gene expression of inflammatory cytokines in the IPEC-J2 cells and maintained the cell integrity. To conclude, SMBS released from Eudragit L100-55 nanofibrous mats in the SIF effectively decreased the adverse effects induced by DON in the IPEC-J2 cells. Nanofibrous mats can release a large amount of SMBS in a short time in SIF to achieve the effect of detoxifying DON.

English

BACKGROUND Amoebic liver abscess is the commonest extra intestinal site of invasive amoebiasis which mainly affects infants and young children. The incidence of pyogenic liver abscess is much higher among children in developing countries than those in developed countries. Diagnosis of liver abscess can be challenging and is often delayed; a high index of suspicion is necessary in children with risk factors. Children have unique set of predisposing causes for liver abscesses. The purpose of this study was to assess the clinical, demographic, and etiological profile of liver abscess in children between 1 month to 12 years of age. METHODS This is a prospective observational study conducted in the Department of Pediatrics, Chacha Nehru Bal Chikistalaya (An Autonomous Institute under Govt. of NCT (National Capital Territory) of Delhi, affiliated to University of Delhi), Delhi from July 2016 to August 2017. All children aged 1 month to 12 years admitted with liver abscess (included consecutively) were enrolled after considering inclusion and exclusion criteria. Written and informed consent was taken from parents/guardians of children aged less than 7 years. Informed assent was taken from children aged more than 7 years, along with written and informed consent from their parents/guardians. Their clinical characteristics, radiological features and laboratory data were analysed. RESULTS Most common age group suffering from liver abscess was 5 - 10 years with male preponderance. Majority of the children belonged to lower socio-economic class and half of them were suffering from malnutrition. Most common clinical presentation of children suffering from liver abscess was fever with pain abdomen and tender hepatomegaly. Majority of the children had leucocytosis, high level of C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). Deranged liver function test with coagulopathy was noted in more than half of the children suffering from liver abscess. Commonest bacterial pathogen was methicillin resistant Staphylococcus aureus followed by Salmonella typhi, Stenotrophomonas maltophilia, coagulase negative Staphylococcus aureus and Staphylococcus hominis. Entamoeba histolytica is a common parasitic agent causing liver abscess in children. CONCLUSIONS Liver abscess should be considered in children presenting with fever and abdominal pain. Most cases involve a single lesion on right lobe of the liver. Methicillin resistant Staphylococcus aureus followed by Salmonella typhi are the two most common pathogens. KEYWORDS Paediatric Liver Abscess, Amoebic Liver Abscess, Pyogenic Liver Abscess, Children

Advancedoral vaccine delivery strategies for improving the immunity.

Infectious diseases continue to inflict a high global disease burden. The consensus is that vaccination is the most effective option against infectious diseases. Oral vaccines have unique advantages in the prevention of global pandemics due to their ease of use, high compliance, low cost, and the ability to induce both systemic and mucosal immune responses. However, challenges of adapting vaccines for oral administration remain significant. Foremost among these are enzymatic and pH-dependent degradation of antigens in the stomach and intestines, the low permeability of mucus barrier, the nonspecific uptake of antigens at the intestinal mucosal site, and the immune suppression result from the elusive immune tolerance mechanisms. Innovative delivery techniques promise great potential for improving the flexibility and efficiency of oral vaccines. A better understanding of the delivery approaches and the immunological mechanisms of oral vaccine delivery systems may provide new scientific insight and tools for developing the next-generation oral vaccine. Here, an overview of the advanced technologies in the field of oral vaccination is proposed, including mucus-penetrating nanoparticle (NP), mucoadhesive delivery vehicles, targeting antigen-presenting cell (APC) nanocarriers and enhanced paracellular delivery strategies and so on. Meanwhile, the mechanisms of delivery vectors interact with mucosal barriers are discussed.

The Development of Ovine Gastric and Intestinal Organoids for Studying Ruminant Host-Pathogen Interactions.

Gastrointestinal (GI) infections in sheep have significant implications for animal health, welfare and productivity, as well as being a source of zoonotic pathogens. Interactions between pathogens and epithelial cells at the mucosal surface play a key role in determining the outcome of GI infections; however, the inaccessibility of the GI tract in vivo significantly limits the ability to study such interactions in detail. We therefore developed ovine epithelial organoids representing physiologically important gastric and intestinal sites of infection, specifically the abomasum (analogous to the stomach in monogastrics) and ileum. We show that both abomasal and ileal organoids form self-organising three-dimensional structures with a single epithelial layer and a central lumen that are stable in culture over serial passage. We performed RNA-seq analysis on abomasal and ileal tissue from multiple animals and on organoids across multiple passages and show the transcript profile of both abomasal and ileal organoids cultured under identical conditions are reflective of the tissue from which they were derived and that the transcript profile in organoids is stable over at least five serial passages. In addition, we demonstrate that the organoids can be successfully cryopreserved and resuscitated, allowing long-term storage of organoid lines, thereby reducing the number of animals required as a source of tissue. We also report the first published observations of a helminth infecting gastric and intestinal organoids by challenge with the sheep parasitic nematode Teladorsagia circumcincta, demonstrating the utility of these organoids for pathogen co-culture experiments. Finally, the polarity in the abomasal and ileal organoids can be inverted to make the apical surface directly accessible to pathogens or their products, here shown by infection of apical-out organoids with the zoonotic enteric bacterial pathogen Salmonella enterica serovar Typhimurium. In summary, we report a simple and reliable in vitro culture system for generation and maintenance of small ruminant intestinal and gastric organoids. In line with 3Rs principals, use of such organoids will reduce and replace animals in host-pathogen research.

Intestinal congestion and reperfusion injury: damage caused to the intestinal tract and distal organs.

In clinical practice, intestinal autologous diseases, ailments and organ transplants can cause severe congestive damage to the intestinal tract. However, after the etiological factor is gotten rid of and blood flow is free without any hinderance, further damage to the intestinal wall often occurs, causing other related organ dysfunctions. This ultimately results in intestinal congestion reperfusion injury (ICRI). When the structure and function of the intestine are destroyed, bacteria, metabolites and endotoxins in the intestinal tract perfuse and enter the portal vein through the already compromised intestinal mucosa, to the other organs via the liver. Nevertheless, this gives rise to further aggravation of the injury, and reperfusion injury syndrome occurs. ICRI is a very common complication encountered by clinicians, and its harm is more severe and serious as compared with that caused by ischemia–reperfusion. Quite a few number of studies on ICRI have been reported to date. The exact mechanism of the injury is still idiopathic, and effective treatment strategies are still limited. Based on recent studies, this article is aimed at reviewing the destruction, damage mechanisms resulting from ICRI to the intestinal anatomical sites and distant organs. It is geared towards providing new ideas for the prevention and therapeutic approaches of ICRI.

Surgery for a small intestine haemorrhage with use of a laparoscope for localization of the bleeding site

Unlike in other parts of the digestive tract, the localization of a haemorrhage site in the small intestine is difficult. In the case of massive bleeding and the need for an urgent operation, localization of the area or site of bleeding in the small bowel may be challenging for the surgeon. The authors report a method of finding sites of bleeding using a laparoscope in a patient with massive gastrointestinal bleeding. Endoscopic diagnostics were inconclusive and an angio-CT revealed bleeding in the small bowel in the left middle-abdomen. Emergency laparotomy was performed with enterotomy and an inspection of the suspected part of the intestine with a laparoscope. Three sites of bleeding were found, of which one with active bleeding was ligated. Additionally, the small intestine parts over the bleeding sites were devascularized by ligation of arterial branches supporting them. This resulted in the bleeding stopping and the patient eventually recovering. The authors believe that the presented technique may be helpful in intraoperative localization of bleeding sites in the small bowel.

Histopathology of human intestinal anastomosis

The faultless seal of the anastomosis is an important aspect of abdominal surgery but opportunities to make a histological evaluation of human intestinal anastomosis sites are rare. This retrospective study examined 30 anastomoses that had been resected following complications or postmortem; the aim was to describe the histological changes at various stages of healing ranging from 4 days to 3.5 years post-surgery. Anastomosis dehiscence showed features of extensive mucosal necrosis and bad submucosal apposition. Old healed anastomoses continued to have chronic inflammatory cells and muscular discontinuity with areas of intervening fibrosis. It would be useful to understand more about how the intraluminal contents are propelled through this scarred area.

Probiotics Modulate Mouse Gut Microbiota and Influence Intestinal Immune and Serotonergic Gene Expression in a Site-Specific Fashion.

Probiotic microorganisms may benefit the host by influencing diverse physiological processes, whose nature and underlying mechanisms are still largely unexplored. Animal models are a unique tool to understand the complexity of the interactions between probiotic microorganisms, the intestinal microbiota, and the host. In this regard, in this pilot study, we compared the effects of 5-day administration of three different probiotic bacterial strains (Bifidobacterium bifidum MIMBb23sg, Lactobacillus helveticus MIMLh5, and Lacticaseibacillus paracasei DG) on three distinct murine intestinal sites (ileum, cecum, and colon). All probiotics preferentially colonized the cecum and colon. In addition, probiotics reduced in the ileum and increased in the cecum and colon the relative abundance of numerous bacterial taxonomic units. MIMBb23sg and DG increased the inducible nitric oxide synthase (iNOS) in the ileum, which is involved in epithelial homeostasis. In addition, MIMBb23sg upregulated cytokine IL-10 in the ileum and downregulated the cyclooxygenase COX-2 in the colon, suggesting an anti-inflammatory/regulatory activity. MIMBb23sg significantly affected the expression of the main gene involved in serotonin synthesis (TPH1) and the gene coding for the serotonin reuptake protein (SERT) in the ileum and colon, suggesting a potential propulsive effect toward the distal part of the gut, whereas the impact of MIMLh5 and DG on serotonergic genes suggested an effect toward motility control. The three probiotics decreased the expression of the permeability marker zonulin in gut distal sites. This preliminary in vivo study demonstrated the safety of the tested probiotic strains and their common ability to modulate the intestinal microbiota. The probiotics affected host gene expression in a strain-specific manner. Notably, the observed effects in the gut were site dependent. This study provides a rationale for investigating the effects of probiotics on the serotonergic system, which is a topic still widely unexplored.

Regulation of Oxygen Homeostasis at the Intestinal Epithelial Barrier Site.

The unique biology of the intestinal epithelial barrier is linked to a low baseline oxygen pressure (pO2), characterised by a high rate of metabolites circulating through the intestinal blood and the presence of a steep oxygen gradient across the epithelial surface. These characteristics require tight regulation of oxygen homeostasis, achieved in part by hypoxia-inducible factor (HIF)-dependent signalling. Furthermore, intestinal epithelial cells (IEC) possess metabolic identities that are reflected in changes in mitochondrial function. In recent years, it has become widely accepted that oxygen metabolism is key to homeostasis at the mucosae. In addition, the gut has a vast and diverse microbial population, the microbiota. Microbiome–gut communication represents a dynamic exchange of mediators produced by bacterial and intestinal metabolism. The microbiome contributes to the maintenance of the hypoxic environment, which is critical for nutrient absorption, intestinal barrier function, and innate and/or adaptive immune responses in the gastrointestinal tract. In this review, we focus on oxygen homeostasis at the epithelial barrier site, how it is regulated by hypoxia and the microbiome, and how oxygen homeostasis at the epithelium is regulated in health and disease.

Pharmacodynamics of Aloe vera and acemannan in therapeutic applications for skin, digestion, and immunomodulation.

Scientific studies of Aloe vera have tentatively explained therapeutic claims from a mechanistic perspective. Furthermore, in vitro outcomes demonstrate that the breakage of acemannan chains into smaller fragments enhances biological effects. These fragments can intravenously boost vaccine efficacy or entrain the immune system to attack cancer cells by mannose receptor agonism of macrophage or dendritic cells. With oral consumption, epithelialisation also occurs at injured sites in the small intestine or colon. The main advantage of dietary acemannan is the attenuation of the digestive process, increasing satiety, and slowing the release of sugars from starches. In the colon, acemannan is digested by microbes into short-chain fatty acids that are absorbed and augment the sensation of satiety and confer a host of other health benefits. In topical applications, an acemannan/chitosan combination accelerates the closure of wounds by promoting granular tissue formation, which creates a barrier between macrophages or neutrophils and the wound dressing. This causes M2 polarisation, reversal of inflammation, and acceleration of the re-epithelialisation process. This review summarises and explains the current pharmacodynamic paradigm in the context of acemannan in topical, oral, and intravenous applications. However, due to contradictory results in the literature, further research is required to provide scientific evidence to confirm or nullify these claims.