Sections Of Intestine Research Articles

Effect of Algae Chlorella extracts for treatment of intestinal tissues of mice infected with Cryptosporidium spp.

Cryptosporidium is a protozoan parasite of medical importance that causes gastroenteritis in a variety of vertebrate hosts, so the study was conducted to evaluate the anti-cryptosporidiosis efficacy of alcoholic and aqueous Chlorella algae extracts in comparison with Azithromycin in the intestines of infected mice. Fecal samples were collected from patients at Al-Kut Hospital suffering from diarrhea in the period December 2021 to end of March 2022, and 90 microscopic samples of both sexes were examined using a modified Ziell-Nelson stain to detect oocysts infected with the parasite. Isolation and flotation purification with Sheather's sugar solution and preservation in potassium dichromate for the purpose of infection in mice. The Experimental study was on groups of 57 mice by dealing with oral parasite oral oocysts within 104 oocyst/ ml except for the negative group addressed by a fishery saline solution. To strict injury, the detailed mice has been examined with microscopic parasites using Ziehl-Nelson Stain, and molecular screening was performed using Multiplex PCR technology. After the mice were divided into five groups with the uninfected and untreated group kept as a healthy negative control. The first group which included 21 mice was treated after it was divided into three subgroups A, B, C for each secondary group 7 mice they were treated with alcoholic extract of Chlorella at different concentrations 50, 100, 150 mg/ml on the respectively, while the second group which included 21 mice on three groups A, B, and C was treated with aqueous extract of Chlorella at the previous concentrations for three consecutive days for each concentration. The third group was treated with azithromycin at a concentration of 500 ml, and the positive control group remained infected with the parasite and was not treated. After treatment a microscopic examination was performed by evaluating the excretion average of parasite oocysts using a hematocytometer slide. The results of histological examination showed that treatment with alcoholic and aqueous Chlorella algae extracts led to remarkable repair and regeneration with restructuring in all sections of the small intestine infected with Cryptosporidium spp. parasite to varying degrees according to the concentrations used. Whereas the groups treated with Chlorella algae extracts showed epithelial layers renewed with the formation of non-enlarged cells and less edema, in addition less infiltration of inflammatory cells was observed in the submucosal layer. There was a slight inflammation in the tissues of the duodenum, jejunum, and ileum with a clear decrease in inflation and the degree of inflammation as well as less severe lesions in the intestinal tissues treated with alcoholic and aqueous extract of Chlorella algae in the highest concentrations with complete disappearance of parasite oocysts.Key words: Intestine, Cryptosporidium spp., Chlorella

An atlas of plasma-accessible carbonic anhydrase availability in the model teleost, the rainbow trout.

The unique teleost oxygenation system that permits enhanced oxygen unloading during stress comprises three main characteristics: pH-sensitive haemoglobin, red blood cell (RBC) intracellular pH (pHi) protection, and a heterogeneous distribution of plasma-accessible carbonic anhydrase (paCA). A heterogeneous distribution of paCA is essential; its presence permits enhanced oxygen unloading during stress, while its absence at the gills maintains conditions for oxygen uptake by pH-sensitive haemoglobins. We hypothesised that paCA would be absent in all four gill arches, as has been previously indicated for arch two, and that paCA would be present in all other tissues. Through a suite of biochemical and molecular methods, we confirmed the absence of paCA from all four arches. We also found evidence for paCA in nine other tissues and a lack of paCA availability in the stomach. Expression was highly variable between tissues and suggests these differences may be associated with their respective metabolic activities. Additionally, we analysed the specific CA-IV isoform expressed within each tissue and showed almost complete separation of expression between tissues; CA-IVa was detected in the heart, brain, anterior intestine, and liver, whereas CA-IVb was detected in all intestine sections, pyloric caeca, kidney, and white muscle. This adds to a growing collection of work suggesting CA-IVa and b play divergent roles in gas exchange and ion/acid-base balance, respectively. The current study represents the first comprehensive atlas of paCA availability within the circulatory system of the model teleost, rainbow trout, and fills important gaps in our knowledge of this unique oxygenation system.

Combined RNAscope and immunohistochemistry staining on duodenal paraffin sections as a new tool to reveal cytolytic potential of intraepithelial lymphocytes

Immunohistochemistry (IHC) is a consolidated technique for the identification of surface and cytoplasmic antigens in cells or tissue sections using specific antibodies, yet simultaneous detection of two markers on the same cell may be difficult to achieve. Here we develop a protocol to perform a double staining using RNAscope, a new in-situ hybridization (ISH) technology, to visualize perforin transcripts, and classical IHC to visualize either CD8 or TcRγδ positive intraepithelial lymphocytes (IELs) in small intestinal paraffin sections of celiac disease (CD) patients. This double assay will allow to investigate the cytotoxic properties of two subsets of IELs in different stages of CD, thus contributing to understand the events leading to tissue destruction and healing.

Active glucose transport varies by small intestinal region and oestrous cycle stage in mice.

What is the central question of this study? Body mass and food intake change during the female ovarian cycle: does glucose transport by the small intestine also vary? What is the main finding and its importance? We have optimised Ussing chamber methodology to measure region-specific active glucose transport in the small intestine of adult C57BL/6 mice. Our study provides the first evidence that jejunal active glucose transport changes during the oestrous cycle in mice, and is higher at pro-oestrus than oestrus. These results demonstrate adaptation in active glucose uptake, concurrent with previously reported changes in food intake. Food intake changes across the ovarian cycle in rodents and humans, with a nadir during the pre-ovulatory phase and a peak during the luteal phase. However, it is unknown whether the rate of intestinal glucose absorption also changes. We therefore mounted small intestinal sections from C57BL/6 female mice (8-9weeks old) in Ussing chambers and measured active ex vivo glucose transport via the change in short-circuit current (∆Isc ) induced by glucose. Tissue viability was confirmed by a positive ∆Isc response to 100µM carbachol following each experiment. Active glucose transport, assessed after addition of 5, 10, 25 or 45mM d-glucose to the mucosal chamber, was highest at 45mM glucose in the distal jejunum compared to duodenum and ileum (P<0.01). Incubation with the sodium-glucose cotransporter 1 (SGLT1) inhibitor phlorizin reduced active glucose transport in a dose-dependent manner in all regions (P<0.01). Active glucose uptake induced by addition of 45mM glucose to the mucosal chamber in the absence or presence of phlorizin was assessed in jejunum at each oestrous cycle stage (n=9-10 mice per stage). Overall, active glucose uptake was lower at oestrus compared to pro-oestrus (P=0.025). This study establishes an ex vivo method to measure region-specific glucose transport in the mouse small intestine. Our results provide the first direct evidence that SGLT1-mediated glucose transport in the jejunum changes across the ovarian cycle. The mechanisms underlying these adaptations in nutrient absorption remain to be elucidated.

Oral supplementation of nicotinamide riboside alters intestinal microbial composition in rats and mice, but not humans

The gut microbiota impacts systemic levels of multiple metabolites including NAD+ precursors through diverse pathways. Nicotinamide riboside (NR) is an NAD+ precursor capable of regulating mammalian cellular metabolism. Some bacterial families express the NR-specific transporter, PnuC. We hypothesized that dietary NR supplementation would modify the gut microbiota across intestinal sections. We determined the effects of 12 weeks of NR supplementation on the microbiota composition of intestinal segments of high-fat diet-fed (HFD) rats. We also explored the effects of 12 weeks of NR supplementation on the gut microbiota in humans and mice. In rats, NR reduced fat mass and tended to decrease body weight. Interestingly, NR increased fat and energy absorption but only in HFD-fed rats. Moreover, 16S rRNA gene sequencing analysis of intestinal and fecal samples revealed an increased abundance of species within Erysipelotrichaceae and Ruminococcaceae families in response to NR. PnuC-positive bacterial strains within these families showed an increased growth rate when supplemented with NR. The abundance of species within the Lachnospiraceae family decreased in response to HFD irrespective of NR. Alpha and beta diversity and bacterial composition of the human fecal microbiota were unaltered by NR, but in mice, the fecal abundance of species within Lachnospiraceae increased while abundances of Parasutterella and Bacteroides dorei species decreased in response to NR. In conclusion, oral NR altered the gut microbiota in rats and mice, but not in humans. In addition, NR attenuated body fat mass gain in rats, and increased fat and energy absorption in the HFD context.

Chlorogenic acid derived from Moringa oleifera leaf as a potential antiinflammatory agent against cryptosporidiosis in mice.

Cryptosporidiosis is a serious illness in immunodeficient patients, and there is still no drug that can completely remove the parasite from the host. The present study represents the first report investigating the impact of the active molecule chlorogenic acid (CGA), naturally isolated from Moringa oleifera leaf extract (EMOLE), on immunosuppressed, Cryptosporidium parvum-infected BALB/c mice. Mice were divided into five groups: normal mice, infected immunosuppressed mice, and infected immunosuppressed mice treated with EMOLE, CGA, and nitazoxanide (NTZ) drugs. Parasitological, immunological, and histopathological investigations were recorded besides differences in the mice' body weight. Infected control mice showed elevated levels of oocyst shedding throughout the study. The EMOLE- and CGA-treated groups showed 84.2% and 91.0% reductions in oocyst shedding, respectively, with no significant difference compared to the drug control. The inflammatory markers IFN-γ, IL-6, IL-1β, and TNF-α were significantly higher in the infected control group. Treatment with 300 mg/kg/day of EMOLE or 30 mg/kg/day of CGA significantly downregulated pro-inflammatory cytokine levels compared to the infected group, although they did not change significantly compared to the NTZ-treated group. Histopathology of intestinal sections showed inflammatory and pathological changes in the infected control group. Low-grade tissue changes and an obvious improvement in villi structure were seen in mice treated with CGA. This study highlighted the role of CGA, isolated and purified from EMOLE, as an effective anti-inflammatory agent in eradicating C. parvum infection.

Screening and Identification of Goat-Milk-Derived Lactic Acid Bacteria with Bacteriocin-like Activity and Probiotic Potentials.

In the current study, we screened 46 isolates of lactic acid bacteria (LAB) derived from goat milk for bacteriocin producers that can inhibit common foodborne pathogens (Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus). The following three strains that showed antimicrobial activity against all indicators were identified: Enterococcus faecalis DH9003 and DH9012, and Lactococcus lactis DH9011. Their antimicrobial products exhibited typical bacteriocin characteristics, such as heat stability and proteinase nature. The bacteriostatic activity of concentrated bacteriocins produced by these LAB was observed at low concentrations (half-minimum inhibitory concentration [MIC50] and 4MIC50), whereas complete inhibition activity against Listeria monocytogenes was detected at high concentrations (16MIC50) of the two Enterococcus faecalis strains (DH9003 and DH9012). Furthermore, the probiotic potentials of the three strains were investigated and described. The results revealed that none of the strains had hemolytic activity, whereas all: were sensitive to ampicillin (50 mg/mL) and streptomycin sulfate (100 mg/mL); were resistant to bile, artificial simulated intestinal tract, and gastric juice at different pH levels (2.5, 3.0, 3.5); and had β-galactosidase activity. Furthermore, all strains exhibited an auto-aggregating phenotype, with self-aggregation ranging from 30% to 55%. DH9003 and DH9012 co-aggregated well with Listeria monocytogenes and Escherichia coli (52.6% and 63.2%, 68.5% and 57.6%, respectively), whereas DH9011 co-aggregated poorly with Listeria monocytogenes (15.6%) and did not co-aggregate with Escherichia coli. Furthermore, our results showed that all three isolates exhibited strong antibacterial activity, tolerance to bile and simulated gastrointestinal environments, adhesion capability, and safety. Finally, DH9003 was selected and used for gavage in rats. By observing the pathological characteristics of rat intestinal and liver tissue sections, DH9003 showed no harmful effects on the intestine and liver of rats, but rather resulted in a denser and longer intestinal mucosa, as well as improving the intestinal mucosa of rats. Considering their substantial prospective applications, we concluded that these three isolates are potential probiotic candidates.

Effect of Algae Spirulina Extraction in treatment of intestinal tissues of mice infected with Cryptosporidium spp.

Cryptosporidiosis caused by Cryptosporidium spp. It is a zoonotic disease that is the most prevalent, so the study was conducted to evaluate the anti-cryptosporidiosis efficacy of alcoholic and aqueous spirulina algae extracts in comparison with Azithromycin in the intestines of infected mice. Fecal samples were collected from patients at Al-Kout Hospital suffering from diarrhea in the period November 2021 to February 2022, and 124 microscopic samples of both sexes were examined using a modified Ziell-Nelson stain to detect eggs infected with the parasite. Isolation and flotation purification with shither sugar solution and preservation in potassium dichromate for the purpose of infection in mice. The Experimental study was on groups of 57 mice by dealing with oral parasite oral oocysts within 104 oocyst/ ml except for the negative group addressed by a fishery saline solution. To strict injury, the detailed mice has been examined with microscopic parasites using Ziehl-Nelson Stain, and molecular screening was performed using Multiplex PCR technology. After the mice were divided into five groups with the uninfected and untreated group kept as a healthy negative control. The first group which included 21 mice was treated after it was divided into three subgroups A, B, C for each secondary group 7 mice they were treated with alcoholic extract of Spirulina at different concentrations 50, 100, 150 mg/ml on the respectively, while the second group which included 21 mice on three groups A, B, and C was treated with aqueous extract of Spirulina at the previous concentrations for three consecutive days for each concentration. The third group was treated with azithromycin at a concentration of 500 ml, and the positive control group remained infected with the parasite and was not treated. After treatment a microscopic examination was performed by evaluating the excretion average of parasite oocysts using a hematocytometer slide. The results of histological examination showed that treatment with alcoholic and aqueous Spirulina algae extracts led to remarkable repair and regeneration with restructuring in all sections of the small intestine infected with Cryptosporidium spp. parasite to varying degrees according to the concentrations used. Whereas the groups treated with Spirulina algae extracts showed epithelial layers renewed with the formation of non-enlarged cells and less edema, in addition less infiltration of inflammatory cells was observed in the submucosal layer. There was a slight inflammation in the tissues of the duodenum, jejunum and ileum with a clear decrease in inflation and the degree of inflammation as well as less severe lesions in the intestinal tissues treated with alcoholic and aqueous extract of Spirulina algae in the highest concentrations with complete disappearance of parasite oocysts.

Therapeutic efficacy of albendazole and berberine loaded on bovine serum albumin nanoparticles on intestinal and muscular phases of experimental trichinellosis

There has been no treatment for trichinellosis until now. Therefore, this work targeted to investigating the efficacy of albendazole and berberine alone and loaded on bovine serum albumin (BSA) nanoparticles against intestinal and muscular phases of trichinellosis in mice. Mice were divided into nine different groups: negative control, positive control, blank nanoparticle, albendazole, berberine, a combination of albendazole and berberine, albendazole-loaded nanoparticle, berberine-loaded nanoparticle and combination of albendazole and berberine-loaded nanoparticle. Subsequently, they were sacrificed 6 and 35 days after infection. Treatment efficacies were parasitologically, histopathologically and, immunohistochemically assessed. Parasitological counting for the adult worms and encysted larvae with histopathological assessment using H&E for intestinal and muscular sections and picrosirius red stain for muscular sections were used. Also, immunohistochemical expression of the intestinal nod-like receptor-pyrin domain containing 3 (NLRP3) was investigated. The group treated with nano_combined drugs showed a statistically significant reduction in adult and encysted larval count (p<0.005), a remarkable improvement of intestinal and muscular inflammation, and a reduction in the capsular thickness of the larvae. Also, this group showed the highest reduction of NLRP3 expression. This work revealed that berberine might be a promising anti-trichinellosis drug with a synergistic effect when combined with albendazole through modulation of the immune response, inflammation, and larva capsule formation. Furthermore, delivering both drugs in a nanoparticle form improves their therapeutic response.

Simultaneous profiling of host expression and microbial abundance by spatial metatranscriptome sequencing.

We developed an analysis pipeline that can extract microbial sequences from spatial transcriptomic (ST) data and assign taxonomic labels, generating a spatial microbial abundance matrix in addition to the default host expression matrix, enabling simultaneous analysis of host expression and microbial distribution. We called the pipeline spatial metatranscriptome (SMT) and applied it on both human and murine intestinal sections and validated the spatial microbial abundance information with alternative assays. Biological insights were gained from these novel data that showed host-microbe interaction at various spatial scales. Finally, we tested experimental modification that can increase microbial capture while preserving host spatial expression quality and, by use of positive controls, quantitatively showed the capture efficiency and recall of our methods. This proof-of-concept work shows the feasibility of SMT analysis and paves the way for further experimental optimization and application.

Poultry By-Product Meal as a Potential Protein Source in Broiler Diets with Exogenous Protease Supplementation

The objective of this study was to investigate the effect of exogenous protease supplementation in diets formulated with poultry by-product meal on growth performance, small intestine magnitudes, and meat physiochemical characters in broiler chickens from 21 to 35 days post-hatch. A total of 120, one-day-old "Arbor Acres Plus" broiler chickens (male: female - 50:50) were allocated one of two dietary treatments to give six replicates and ten birds per cage. Two dietary treatments included a control diet (CON) and a diet supplemented with protease (CON+Pro). At day 35, body weight and feed intakes were measured to calculate the feed conversion ratio for the entire experiment period. Two birds from each pen were euthanized to measure the relative lengths and relative weights of three small intestine sections and meat samples were collected for physiochemical characteristic analyses at 35 days post-hatch. Exogenous protease supplementation did not influence (<i>P</i>&gt;0.05) growth performance but showed a tendency to improve FCR (<i>P</i>=0.082). Protease supplementation showed a tendency to reduce proximal small intestine length (<i>P</i>=0.091). Broilers offered dietary treatments influenced minced meat color where protease supplementation resulted in lower CIE b^* (<i>P</i>&lt;0.001) colorimetric value for yellowness and showed a significant trend (<i>P</i>=0.059) on reducing meat redness CIE a^*. In conclusion, the addition of exogenous protease to a broiler diet formulated with poultry by-product meal did not affect the growth performance, small intestine magnitudes, and meat physiochemical parameters (except CIE b^*) in broiler chickens.

Domestic researchers who significantly contributed in the study of digestive system diseases: images in art. Vasyl Parmenovich Obraztsov

Vasyl Parmenovich Obraztsov (1849 — 1920) was an outstanding therapist, Professor of Medicine, and innovator in the field of techniques for diagnosing diseases of the cardiovascular and digestive systems. The research interests of V. P. Obraztsov were mainly focused on studying the most important issues in general and specific gastroenterology and cardiology. At the same time, he aimed to invent reliable techniques for the physical diagnosis of diseases of the abdominal organs and heart.&#x0D; Palpation of the abdominal organs, invented and introduced into clinical practice by V. P. Obraztsov, is based on four principles. During deep breathing, especially when the patient is lying down, the abdominal press occasionally relaxes. During the exhalation phase, the researcher tries to reach deep into the abdominal cavity with a palpating hand and press an organ that needs to be felt against the posterior abdominal wall. The first principle of the research technique that V. P. Obraztsov came up with is deep palpation. It is usually possible to palpate any organ in the abdomen when the fingertips of the palpating hand slide off the edge of the organ being examined. The palpable part of the gastrointestinal tract is pressed against the posterior abdominal wall with a strong downward movement of the hand that is perpendicular to the organ’s long axis. When the fingers slip from the organ, the localization of the organ, as well as its consistency, shape, width, and mobility, can be determined; at the same time, the patient may report unpleasant or painful sensations. Sliding palpation is the second principle of the research technique by V. P. Obraztsov. According to topographic anatomy, each organ in the abdominal cavity has its own normal localization. Now, X‑rays confirm this, but V. P. Obraztsov had to do experiments to develop a typical topography of certain segments of the digestive tract in the abdominal cavity of a healthy person and find typical projections of internal organs on the anterior wall of the abdomen. Further studies by his students proved the correctness of his description of the topography of the abdominal organs. Of course, both V. P. Obraztsov and his students repeatedly emphasized the possibility of individual differences or physiological variants in the localization of the digestive tract organs in patients. Topographic palpation has become the third guiding principle of the research technique developed by V. P. Obraztsov. V. P. Obraztsov suggested that a study of the organs in the abdomen be done in a certain sequence. First, the sigmoid colon is examined, then the cecum, the terminal segment of the small intestine, the appendix, the stomach, with an attempt to palpate its small and large curvatures, and the pylorus. After that, the palpation of the transverse colon and the ascending and descending sections of the large intestine starts. The examination is completed by determining the localization, consistency, shape, mobility, and other features of the parenchymal organs — the liver, spleen, and kidneys. Thus, systematic palpation is the fourth principle of the examination of the abdominal organs.&#x0D; V. P. Obraztsov made a significant contribution to the history of cardiology. Systematic cardiac auscultation with a stethoscope and directly with the ear let him describe some features of the tone I, report the presence of a normal tone III, and point out the relatively frequent detection of bifurcation and splitting of heart tones. A series of works by V. P. Obraztsov on the study of the gallop rhythm is of particular importance for cardiology.&#x0D; In 1950, on the centenary of his birth, the government of the Ukrainian SSR decided to publish his «Selected Works», build a monument, and assign his name to the Department of Internal Diseases No. 1 of the Kyiv Medical Institute. By the same date, the country’s medical journals had published his students’ memoirs, as well as articles and essays about his life path.&#x0D;

MUC-FIRE: Study protocol for a randomized multicenter open-label controlled trial to show that MUCous FIstula REfeeding reduces the time from enterostomy closure to full enteral feeds

BackgroundAfter enterostomy creation, the distal bowel to the ostomy is excluded from the physiologic passage of stool, nutrient uptake, and growth of this intestinal section. Those infants frequently require long-term parenteral nutrition, continued after enterostomy reversal due to the notable diameter discrepancy of the proximal and distal bowel. Previous studies have shown that mucous fistula refeeding (MFR) results in faster weight gain in infants. The aim of the randomized multicenter open-label controlled MUCous FIstula REfeeding (“MUC-FIRE”) trial is to demonstrate that MFR between enterostomy creation and reversal reduces the time to full enteral feeds after enterostomy closure compared to controls, resulting in shorter hospital stay and less adverse effects of parenteral nutrition.Methods/Design: A total of 120 infants will be included in the MUC-FIRE trial. Following enterostomy creation, infants will be randomized to either an intervention or a non-intervention group.In the intervention group, perioperative MFR between enterostomy creation and reversal will be performed. The control group receives standard care without MFR.The primary efficacy endpoint of the study is the time to full enteral feeds. Secondary endpoints include first postoperative bowel movement after stoma reversal, postoperative weight gain, and days of postoperative parenteral nutrition. In addition adverse events will be analyzed. DiscussionThe MUC-FIRE trial will be the first prospective randomized trial to investigate the benefits and disadvantages of MFR in infants. The results of the trial are expected to provide an evidence-based foundation for guidelines in pediatric surgical centers worldwide. Trial registrationThe trial has been registered at clinicaltrials.gov (number: NCT03469609, date of registration: March 19, 2018; last update: January 20, 2023, https://clinicaltrials.gov/ct2/show/NCT03469609?term=NCT03469609&draw=2&rank=1).

Acrylamide-Induced Changes in the Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) Immunoreactivity in Small Intestinal Intramural Neurons in Pigs.

A particularly pressing problem is determining consumer-safe doses of potentially health- and life-threatening substances, such as acrylamide. The aim of the study was to determine how acrylamide affects the pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive intramural neurons in the small intestine of sexually immature gilts. The study was conducted on 15 sexually immature Danish gilts receiving for 28 days empty gelatin capsules or acrylamide in low (0.5 µg/kg of body weight (b.w.)/day) and high (5 µg/kg b.w./day) doses. After euthanasia, intestinal sections were stained using the double immunofluorescence staining procedure. Studies have shown that oral administration of acrylamide in both doses induced a response of intramural neurons expressed as an increase in the population of PACAP-immunoreactive neurons in the small intestine. In the duodenum, only in the myenteric plexus (MP) was an increase in the number of PACAP-immunoreactive (IR) neurons observed in both experimental groups, while in the outer submucous plexus (OSP) and inner submucous plexus (ISP), an increase was noted only in the high-dose group. In the jejunum, both doses of acrylamide led to an increase in the population of PACAP-IR neurons in each enteric plexus (MP, OSP, ISP), while in the ileum, only supplementation with the higher dose of acrylamide increased the number of PACAP-IR enteric neurons in the MP, OSP, and ISP. The obtained results suggest the participation of PACAP in acrylamide-induced plasticity of enteric neurons, which may be an important line of defence from the harmful action of acrylamide on the small intestines.

Potential determinants of low circulating glucagon-like peptide 2 concentrations in Zambian children with non-responsive stunting.

What is the central question of this study? Non-responsive stunting is characterised by a progressive decline of circulating glucagon-like peptide 2: what are the possible causes of this decline? What is the main finding and its importance? In contrast with the established loss of Paneth and goblet cells in environmental enteropathy, there was no evidence of a parallel loss of enteroendocrine cells as seen by positive tissue staining for chromogranin A. Transcriptomic and genomic analyses showed evidence of genetic transcripts that could account for some of the variability seen in circulating glucagon-like peptide 2 values. Nutrient sensing determines digestive and hormonal responses following nutrient ingestion. We have previously reported decreased levels of glucagon-like peptide 2 (GLP-2) in children with stunting. Here we demonstrate the presence of enteroendocrine cells in stunted children and explore potential pathways that may be involved in reduced circulating levels of GLP-2. At the time of performing diagnostic endoscopies for non-responsive stunted children, intestinal biopsies were collected for immunofluorescence staining of enteroendocrine cells and transcriptomic analysis. Circulating levels of GLP-2 were also measured and correlated with transcriptomic data. An exploratory genome-wide association study (GWAS) was conducted on DNA samples (n=158) to assess genetic contribution to GLP-2 variability. Intestinal tissue sections collected from non-responsive stunted children stained positive for chromogranin A (88/89), alongside G-protein-coupled receptors G-protein receptor 119 (75/87), free fatty acid receptor 3 (76/89) and taste 1 receptor 1 (39/45). Transcriptomic analysis found three pathways correlated with circulating GLP-2: sugar metabolism, epithelial transport, and barrier function, which likely reflect downstream events following receptor-ligand interaction. GWAS analysis revealed potential genetic contributions to GLP-2 half-life and receptor binding. Enteroendocrine cell loss was not identified in stunted Zambian children as has been observed for goblet and Paneth cells. Transcriptomic analysis suggests that GLP-2 has pleiotrophic actions on the intestinal mucosa in malnutrition, but further work is needed to dissect pathways leading to perturbations in nutrient sensing.

Correlation of extracellular enzymes activity of Candida glabrata clinical isolates with in vivo pathogenicity in Galleria mellonella larvae

Objective: To explore the relationship between extracellular enzymes activity and virulence of Candida glabrata clinical isolates based on the infection model of Galleria mellonella larvae. Methods: Using experimental research methods, 71 strains of non-repetitive Candida glabrata were collected from Qinghai Provincial People's Hospital from June 2021 to January 2022. Bovine serum protein agar medium, egg yolk agar medium, sheep blood agar medium, Tween-80 agar medium and triglyceride agar medium were used to detect the aspartyl protease activity, phospholipase activity, hemolysis activity, esterase activity and lipase activity of Candida glabrata. Median lethal concentration (LC50) was calculated by using 1.25×108 CFU/ml,2.50×108 CFU/ml,3.75×108 CFU/ml,5.00×108 CFU/ml suspension of Candida glabrata ATCC2001 to infect Galleria mellonella larvae. Histopathological and etiological analysis was performed to determine whether the infection model was successfully established. The clinical isolates of Candida glabrata were configured to infect Galleria mellonella larvae with LC50 concentration to detect the pathogenicity of Galleria mellonella larvae.Spearman test or Pearson test were used to analyze the correlation between the extracellular enzyme activity of Candida glabrata clinical isolates and the pathogenicity of Galleria mellonella larvae. Results: 71 strains of Candida glabrata isolated clinically were detected to have low hemolytic activity after 2 days of culture. Aspartyl protease was detected after 4 days of culture, among which 7 strains (9.86%), 19 strains (26.76%) and 45 strains (63.38%) showed low, medium and high aspartyl protease activity. After 7 days of culture, 71 strains did not detect phospholipase, esterase and lipase activities. Candida glabrata on Galleria mellonella larvae of LC50=2.5×108 CFU/ml Fungal spore were found in the intestinal tissue pathological section of Galleria mellonella larvae in the experimental group, and Candida glabrata was identified by the microbial Mass Spectrometry after culture, while no fungi were found in the pathological section and culture of the control group. Spearman test shows that, there was a linear positive correlation between aspartyl protease activity and the survival rate of Galleria mellonella larvae (r = 0.73, P<0.01), the difference was statistically significant.Pearson test shows that, there was no significant linear relationship between hemolytic activity and survival rate of Galleria mellonella larvae (r = 0.16, P = 0.34), the difference was not statistically significant. Conclusion: The clinical isolates of Candida glabrata in this study had aspartyl protease activity and low hemolytic activity, but no phospholipase, esterase and lipase activity. The activity of aspartyl aspartyl protease of Candida glabrata was positively correlated with the pathogenicity of Galleria mellonella larvae.

Long-Term Complications of Neonatal Necrotic Enterocolitis: Twisting of Stenosed Small Intestine

Neonatal necrotic enterocolitis (NEC) is a nonspecific inflammatory disease of unknown etiology with multifactorial pathogenesis, which development is explained as a result of hypoperfusion of the immature intestinal mucosa of a newborn who has undergone perinatal hypoxia and, as a result, a change in blood flow in the mesenteral vascular system. This disease more often affects the intestinal wall of premature babies. In typical cases, mucosal necrosis develops in the terminal iliac and right parts of the colon, and when the process progresses, it can spread to the entire thickness of the intestinal wall, causing its perforation, so peritonitis is a frequent complication of NEC. But in addition, distant complications of NEC are distinguished, which include the development of intestinal obstruction in view of a decrease in the contractility of the intestinal wall section due to its fibrosis and, as a result, narrowing of the lumen.We report the clinical case of surgical treatment of the twisting of stenosed portion of the ileum in a child who underwent NEC in the early neonatal period. A mother sought assistance with a child of 3 months at the Reginal Children’s Hospital (born at 28 weeks of gestation). After birth, there was a violation of the absorption of enteral nutrition (periodic posseting, bloating), blood in the stool was determined. On the 14th day of life a pediatric surgeon examined the baby: necrotizing enterocolitis II A. After stabilizing the condition (2 months), the child was transferred from the perinatal center to a pediatric hospital with a diagnosis of bronchopulmonary dysplasia, a new form, a severe course, and a period of exacerbation. At the age of 2.5 months, he was discharged in a satisfactory condition to the outpatient stage. On the 15th day from the moment of discharge (3 months of life), the mother noted the expressed anxiety of the baby, bloating, stool retention. On the 16th day from the moment of discharge during feeding, the child began to suck sluggishly, did not absorb the age norm. He was examined by a pediatrician, the consultation of a pediatric surgeon was recommended. Due to the severity of the condition, the baby was hospitalized.In the initial examination, no data for intestinal obstruction were detected. The patient received parenteral nutrition, antibacterial therapy. Ultrasound and radiography of abdominal organs were performed daily. On the third day of observation, deterioration is noted (abdomen bloated mainly in the upper parts, soft upon with palpation, the baby was anxious, peristalsis was reduced), with ultrasound: between the loops of the enlarged intestine, an echogenic band of up to 18 mm (adhesion?) was determined, there was the liquid component between the loops. A laparotomy was performed, during the revision there was a displacement of the large intestine into the left parts of the abdominal cavity. Ten cm from the ileocecal angle, a section of the ileum with a length of up to 15 cm was found, which was like a “double trunk” wrapped at the base around its axis around a cord-like adhesive stretching to the posterior abdominal wall. Visually, the loop was dark bard in color, its walls were swollen, infiltrated, and their cartilaginous density was determined upon palpation in the contact area of the walls of the intestine. The resection of this loop was performed, end-to-end anastomosis was formed according to the method of J. Louw. After the operation, the baby received treatment in the intensive care unit, enteral feeding on day 5, on day 9, after expanding the volume of feeding, he was transferred to the department of pediatric surgery. Discharged in satisfactory condition on the 12th day after surgery.

ЛИПОМА КАК СЛУЧАЙНАЯ НАХОДКА В АБДОМИНАЛЬНОЙ ХИРУРГИИ

Primary neoplasms of the abdominal cavity are rare and usually have a mesenchymal origin. Lipoma is a benign mature tumor of adipose tissue, it occurs more often in women aged 30 to 50 years. Retroperitoneal lipoma is a rare variant of lipoma, manifested by various signs and symptoms that can mislead specialists. Lipomas are more common in abdominal surgery in the gastrointestinal tract. In this localization, the main pathognomonic complication is intestinal intussusception. Diagnosis is difficult, due to the relatively slow growth of education, characteristic clinical manifestations may be absent for a long time. X-ray imaging is the preferred diagnostic method. To date, surgical resection is the priority method of treatment of this pathology. The analysis of scientific literature on the occurrence of abdominal lipomas is carried out. The diagnostic significance in assessing the condition and complications of lipomas has been studied. Clinical cases from personal practice on the basis of the State Clinical Hospital named after him are presented. Yudina. Patients with diagnoses of giant retroperitoneal lipoma and complicated small intestine lipoma by female intussusception aged 62 and 78 years, respectively. The first clinical case demonstrates the randomness of the determination of lipoma in a patient admitted to the clinic with a closed craniocerebral injury and a wound of the soft tissues of the head. Ultrasound of the abdominal cavity showed the presence of a large retroperitoneal formation of an inhomogeneous consistency. During laparotomy, a giant fatty tumor with clear boundaries was revealed, closely adjacent to the adipose tissue of the retroperitoneal space. Lipoma was removed, drainage of the abdominal cavity and retroperitoneal space was performed. Histopathological examination confirmed the diagnosis of retroperitoneal lipoma. The weight of the lipoma was 10.610 kg. The postoperative period proceeded without complications. The second clinical case shows how a long-term undiagnosed lipoma of the gastrointestinal tract leads to a complicated course, namely intussusception. Patient A. 78 years old with discomfort and abdominal pain during the last few weeks, small intestine invagination was detected using ultrasound. Diagnosis upon admission: acute pancreatitis. A resection of the altered section of the small intestine was performed. The postoperative period was favorable. The patient was discharged on the 10th day. The presented clinical cases of this disease once again confirm the importance of timely dispensary diagnosis of abdominal tumors.

P068 Polyunsaturated fatty acid excess fuels pro-inflammatory transcriptional networks in GPX4 deficient intestinal epithelial cells

Abstract Background There is increasing evidence that the rise in Crohn’s disease (CD) incidence is partially driven by constituents of a Western-style diet such as polyunsaturated fatty acids (PUFA). Recently we could demonstrate that PUFA excess induces a CD-like enteritis in mice lacking antioxidant GPX4 in their intestinal epithelial cells (IECs) and that CD patients clinical disease activity positively correlates with PUFA intake. In this project we investigated the role of epithelial lipid composition and lipid mediators during PUFA induced metabolic gut inflammation. Furthermore, we transcriptionally profiled GPX4 deficient small intestinal cells after stimulation with ω3- and ω6-PUFA. Methods Mice lacking one allele of Gpx4 in their IECs (GPX4+/-IEC) and wild-type controls (WT) were fed a PUFA (10% fish oil) enriched Western diet (PUFA-WD) for 12 weeks. Intestinal inflammation was assessed by histopathology. IECs were harvested by scraping of small intestinal sections and lipid profiles and mediators were analysed with LC/MS. GPX4 in murine small intestinal cells (MODE-K) was knocked down by siRNA (siGpx4) scrambled siRNA served as control (siCtrl). MODE-K were stimulated with ω3-, ω6-PUFA, 5,6- and 8,9-DHET or vehicle. Cytokine expression in cell culture supernatant was analysed by ELISA and transcriptome was analysed by bulk RNA sequencing. Results Histopathology revealed gut inflammation in GPX4+/-IEC but not WT mice after 12 weeks PUFA-WD feeding. LC/MS analysis of IECs derived from GPX4+/-IEC and WT mice fed the PUFA-WD depicted a significant increase in eicosapentaenoic acid (20:5), docosapentaenoic acid (22:5) and docosahexaenoic acid (22:6) containing phosphatidylethanolamine (PE) and phosphatidylcholine (PC) species (Fig 1a). All other PE and PC species showed a reduced amount, confirming the enrichment of the specific PUFAs within the IECs by the PUFA-WD. Lipid mediator analysis further revealed an increase in 5,6-DHET and 8,9-DHET only in IECs derived from GPX4+/-IEC but not WT mice (Fig 1b). When stimulated with PUFA, siGpx4 MODE-K produced high amounts of neutrophil attracting chemokine CXCL-1 which is attenuated by the supplementation of anti-inflammatory lipid mediators 5,6-DHET and 8,9-DHET (Fig 2). Transcriptome analysis revealed the upregulation of pro-inflammatory pathways including TNFa, MAPK and NF-κB in siGpx4 vs. siCtrl MODE-K after PUFA stimulation (Fig 3). Conclusion By integrating lipidomic and transcriptomic data of a reductionist cell culture system and a model organism, we could show that PUFA excess in a GPX4 deficient individual fuels a pro-inflammatory transcriptional network which outweighs anti-inflammatory ω3-PUFA metabolites influence culminating in gut inflammation.

Impact of a 24 h feed withdrawal on active nutrient transport, intestinal morphology and gene expression in the equine small and large intestine

Abstract Horses are often subjected to short term feed withdrawal (FW) pre- or post-surgery to reduce anesthetic complications. However, removing nutrients from the intestinal lumen may negatively impact intestinal health. Thirteen horses were used to determine the effects of a 24 h FW on gut barrier function, active nutrient transport, transporter gene expression and intestinal morphology. Following 0 or 24 h FW (0FW or 24FW, respectively), horses were euthanized via overdose of sodium- pentobarbital and –phenytoin, and segments of proximal jejunum (PJ), mid jejunum (MJ), ileum (Il) and right ventral colon (RVC) were harvested for histology (PJ and Il), gene expression, and active nutrient transport analysis. Active transport measurements were determined using modified Ussing chambers following the addition of glucose, phosphorus, glutamine and Gly-Sar. Carbachol induced chloride (Cl) ion secretion was measured to examine the diarrhetic response. Messanger RNA expression of the intestinal Na-dependent glucose cotransporter (SGLT1), fructose transporter (GLUT5), di- and tri-peptide transporter (PepT1) and neutral AA/glutamine transporter (ASCT2) were determined using RT-PCR. The GLM procedure of SAS was used to determine the effects of FW and responses among various intestinal sections. The horse served as the experimental unit. Villus heights (P &amp;lt; 0.002) and crypt depths (P &amp;lt; 0.02) in the Il were larger than in the PJ, though no differences were observed between 0FW and 24FW horses. Active glutamine absorption increased 82% in the PJ of 24FW horses compared to 0FW horses (P &amp;lt; 0.02). The mRNA expression of SGLT1 decreased (P &amp;lt; 0.05) moving aborally in the gastrointestinal tract. Horses subjected to 24FW had 82% less GLUT5 (P &amp;lt; 0.05) and 61% less PepT1 mRNA expression in the PJ, compared to 0FW horses. Interestingly, ASCT2 mRNA expression increased 164% from PJ to RVC (P = 0.05). However, a 36% decrease in ASCT2 mRNA expression was observed overall for 24FW horses. This data indicates that SGLT1, GLUT5, PepT1 and ASCT2 are expressed throughout the small intestine and RVC of the horse at varying concentrations, and that they can be differentially regulated by a 24 h FW. Data from this experiment also indicates that a 24 h FW results in up regulation of active glutamine absorption, presumably in an effort to supply glutamine as an energy substrate for enterocytes.