Number Of Intraepithelial Lymphocytes Research Articles

The Role of Zinc, Inflammation and Duodenal Mucosal Integrity in Cholesterol Gallstone Diseases

Introduction: Cholesterol Gallstone (GSD) is a common multifactorial disease characterized by an aggregation and growth of cholesterol crystals in the gallbladder. The global prevalence of GSD is ˜10-20% in adult population, but in Chile it rises to 28%, 17% men and 30% women, respectively. The small intestine have been identified contribute of developing gallstone, however, the molecular mechanism of the small intestine in the GSD pathogenesis have been poorly studied. Therefore, our aim was to identify the molecular mechanism of small intestine in GSD subject. Methods: Duodenal biopsy samples were obtained from patients with GSD and healthy volunteers. Presence or absence GSD was defined by abdominal ultrasonography. We performed transcriptome studies using the Illumina HiSeq 2500. We used real time PCR and immunofluorescence to validate the differentially expressed genes identified in GSD subjects. For identify number of intraepithelial lymphocytes and lysozymes expression were assessed by immunohistochemistry and immunofluorescence, respectively. Results: We identified 548 differentially expressed genes in GSD compared with control subjects. We observed enriched biological process related to cellular response to zinc, immune and antimicrobial responses, epithelial tube morphogenesis, apoptosis, DNA replication, and phospholipid metabolic process. Patients with GSD showed lower metallothionein and NPC1L1 expression, zinc level and tight junction gene expression, in change, an increased in immune and antimicrobial genes. A significant association between the zinc level and phytosterol concentration were observed. Conclusion: We identify for the first time that upper small intestine of GSD subjects have altered the zinc level, immune and antimicrobial responses and duodenal barrier function. These evidences allow us to hypothesize that the small intestine may have a key impact in GSD pathogenesis. Acknowledgment: Funded by FONDECYT 3160400. FONDAP Center for Genome Regulation 15090007.

Is revision of cutoff values needed when using CD3 immunohistochemical staining in histopathologic diagnosis of lymphocytic colitis?

Lymphocytic colitis (LC) and LC incomplete (LCi) are common causes of chronic watery diarrhea. The diagnosis relies on clinical findings and histopathologic evaluation. The diagnostic criteria of LC are based on hematoxylin and eosin (HE) staining. However, supplementary immunohistochemical staining for highlighting the lymphocytes in borderline cases is now widely used. This change in diagnostics could lead to incorrectly diagnosing patients with LC and LCi if the present histologic criteria are used. The number of intraepithelial lymphocytes (IELs) was estimated and categorized in intervals based on HE- versus CD3-stained slides from patients with an HE diagnosis of normal colonic mucosa (n = 19), mucosa with nonspecific reactive changes (n = 24), LCi (n = 24), and LC (n = 40). The number of IELs was compared with clinical symptoms. Overall, the number of IELs was higher with CD3 stain compared with HE stain in 73% of cases, unchanged in 26% of cases, and lower in 1 case. The number of IELs detected was higher using the CD3 stain in 53%, 79%, 79%, and 75% of cases included as normal colonic mucosa, nonspecific reactive changes, LCi, and LC, respectively. Based on CD3 stain, 58% of the cases with nonspecific reactive changes fulfilled the HE criteria for LCi, and 79% of the cases with LCi fulfilled the HE criteria for LC. Automated image analysis of CD3-stained slides resulted in even higher numbers of IELs in all 4 diagnostic groups. Conclusively, our data support considering increased cutoff values for LCi and LC when assessed in CD3-stained specimens.

The effects of the dietary supplementation of Echinacea purpurea extract and/or vitamin C on the intestinal histomorphology, phagocytic activity, and gene expression of the Nile tilapia

In this study, the influence of the dietary incorporation of Echinacea purpurea (EP) extract and/or vitamin C on the intestinal histomorphology and some immunological indices were tested in the Nile tilapia (Oreochromis niloticus Linn.). O. niloticus were randomly divided into four groups. The control group G1 was fed on a basal diet, while the G2 and G3 were fed on basal diets, supplemented with EP extract and vitamin C at the doses of 500 mg kg−1 and 400 mg kg−1, respectively. Meanwhile, G4 was fed on a basal diet, supplemented with a mixture of EP extract and vitamin C. After 28 days of feeding, the intestinal tissues were collected for histological observation and immune status, was based on an assay for measuring the phagocytic activity. Furthermore, the expression of the transforming growth factor-beta 1 (TGF-β1), interleukin-1beta (IL-1β), and tumor necrosis factor alpha (TNF-α) genes was evaluated in intestine and head kidney. The results revealed that the G4 successfully surpassed the other groups in terms of the heights of intestinal villi, the number of goblet cells and intraepithelial lymphocytes (IELs), and the phagocytic activity, followed by the G3 and G2. The expression of the IL-1β and TNF-α genes were up regulated only in G4 but in the G3 only the expression of the IL-1β gene was up regulated. Hence, EP extract along with vitamin C could be used as a feed additive in order to improve the structure of the intestinal mucosal epithelium and immune response in tilapia.

Insights of Leishmania (Viannia) braziliensis infection in golden hamster (Mesocricetus auratus) intestine

AimThe present study compared and evaluated morphological and quantitative alterations in the ileum of hamsters infected by two L. (V.) braziliensis strains isolated from patients with different lesion aspects and treatment responses. Main methodsHamsters were infected in the left hindpaw with a suspension of promastigotes (2 × 107/100 μl) of two different strains of L. (V.) braziliensis. After 90 or 120 days, the animals were euthanized. Samples of the ileum and mesenteric lymph node were collected for histological examination and quantitative polymerase chain reaction. Key findingsAll infected animals developed similar profile of paw lesions. In peripheral blood there was an increase in the number of mononuclear cells which contributed to elevated global leukocytes count. Increases in the width and height of villi and width and depth of crypts were observed. The thickness of the muscular layers, submucosa, and intestinal wall also increased. Histopathological alterations were observed, including inflammatory infiltrate in crypts and a large number of immune cells in the lamina propria, submucosa, and muscular layer. Immune cells were found inside myenteric ganglia, with an increase in the number of intraepithelial lymphocytes. Leishmania DNA was detected in the ileum and mesenteric lymph node at both times of infection. The presence of amastigotes in the ileum was revealed by immunohistochemistry. SignificanceThe infection with different strains of L. (V.) braziliensis causes morphological and quantitative alterations in the ileum of hamsters and the parasite can migrate to the mesenteric lymph node and intestine.

Functional intraepithelial lymphocyte changes in inflammatory bowel disease and spondyloarthritis have disease specific correlations with intestinal microbiota

BackgroundDysbiosis occurs in spondyloarthritis (SpA) and inflammatory bowel disease (IBD), which is subdivided into Crohn’s disease (CD) and ulcerative colitis (UC). The immunologic consequences of alterations in microbiota, however, have not been defined. Intraepithelial lymphocytes (IELs) are T cells within the intestinal epithelium that are in close contact with bacteria and are likely to be modulated by changes in microbiota. We examined differences in human gut-associated bacteria and tested correlation with functional changes in IELs in patients with axial SpA (axSpA), CD, or UC, and in controls.MethodsWe conducted a case-control study to evaluate IELs from pinch biopsies of grossly normal colonic tissue from subjects with biopsy-proven CD or UC, axSpA fulfilling Assessment of SpondyloArthritis International Society (ASAS) criteria and from controls during endoscopy. IELs were harvested and characterized by flow cytometry for cell surface markers. Secreted cytokines were measured by ELISA. Microbiome analysis was by 16S rRNA gene sequencing from rectal swabs. Statistical analyses were performed with the Kruskal-Wallis and Spearman’s rank tests.ResultsThe total number of IELs was significantly decreased in subjects with axSpA compared to those with IBD and controls, likely due to a decrease in TCRβ+ IELs. We found strong, significant negative correlation between peripheral lymphocyte count and IEL number. IELs secreted significantly increased IL-1β in patients with UC, significantly increased IL-17A and IFN-γ in patients with CD, and significantly increased TNF-α in patients with CD and axSpA as compared to other cohorts. For each disease subtype, IELs and IEL-produced cytokines were positively and negatively correlated with the relative abundance of multiple bacterial taxa.ConclusionsOur data indicate differences in IEL function among subjects with axSpA, CD, and UC compared to healthy controls. We propose that the observed correlation between altered microbiota and IEL function in these populations are relevant to the pathogenesis of axSpA and IBD, and discuss possible mechanisms.Trial registrationClinicalTrials.gov, NCT02389075. Registered on 17 March 2015.

Does the number of mucosal immune cells differ in irritable bowel syndrome and its subtypes?

Recently, mucosal inflammation has been proposed to be one of the mechanisms underlying the pathophysiology of irritable bowel syndrome (IBS); however, there are controversial results regarding this hypotheses. Our aim was to evaluate immune cell infiltration in rectal and ileal biopsy specimens of patients with IBS and to compare it with those of healthy controls. In total, 36 patients with IBS (15 with diarrhea and 21 with constipation) and 16 healthy volunteers were enrolled. Ileocolonoscopy and ileal/rectal biopsies were performed. Rectal and terminal ileal biopsy specimens were evaluated for mucosal immune cell infiltration using immunohistochemical analysis. Serotonin positivity as well as counts of intraepithelial lymphocytes (IEL) and CD4+, CD8+, CD20+, and CD3+ cells were determined by a single pathologist who is an expert in the gastrointestinal system. CD3+ and CD4+ cell counts in rectal and terminal ileal biopsy specimens were lower in the IBS group than in the controls. Conversely, there was no statistically significant difference between the IBS and control groups in terms of serotonin positivity as well as counts of IEL and CD20+ and CD8+ cells. Comparison between the IBS subgroups revealed a higher number of IEL in rectal biopsy specimens of the diarrhea dominant group. In the IBS subgroups, immune cell counts in terminal ileal and rectal biopsy specimens showed a positive correlation. IBS and its subgroups showed lower immune cell counts than the controls in our study. These results indicate that there is no significant mucosal inflammation in homogeneous groups of patients with IBS. Rectal biopsies may be sufficient for the evaluation of inflammation in IBS.

Quantitation of Colonic Cells as Severity Markers in Patients with Irritable Bowel Syndrome

Background: Irritable bowel syndrome (IBS) is the most common gastrointestinal syndrome. Routine histopathology and immunohistochemistry (IHC) evaluations have shown an increase in the number of different inflammatory cells in the colon of IBS patients. In this study, we have compared the number of intraepithelial lymphocytes (IELs), eosinophils, mast cells and CD3+ T cells, in IBS patients and normal subjects. Materials and Methods: In 2016, seventy-nine patients with IBS and seventy-nine healthy subjects who underwent colonoscopy for other non-specific causes and with no pathologic findings, were enrolled in this cross-sectional study. Biopsy specimens obtained from the colon were stained, using IHC methods to determine the number of IELs, eosinophils, mast cells and CD3+ T cells. Quantitative and qualitative variables were compared between the two groups, using a Chi-square test and Student’s t-test. Results: Seventy-nine patients with IBS, 79.7% females with a mean age of 42.5±14.6 years, were recruited, as the case group, and seventy-nine individuals, 51.9% females with a mean age of 39.7±18.9 years, were enrolled as controls. The average number of IELs per high power fields (hpf) was found to be higher in the IBS group, and this difference was statistically significant (32.8±11.8 vs. 28.6±12.9; P=0.034). Also, the mean count/hpf of CD3+ T lymphocytes (23.1±7.9 vs. 20.2±8.1; P=0.024) and mast cells (7.6±3.1 vs. 6.6±3.0; P=0.041) were significantly higher in the IBS group, compared to the control group. The number of eosinophils was higher in the IBS group, but the differences were not statistically significant (P=0.066). Conclusion: According to the results, we suggest that analysis of immune cells and IELs in intestinal biopsies might be an appropriate method for diagnosis of IBS. [GMJ.2018;7:e1063]

Quantitation of Colonic Cells as Severity Markers in Patients with Irritable Bowel Syndrome

Background: Irritable bowel syndrome (IBS) is the most common gastrointestinal syndrome. Routine histopathology and immunohistochemistry (IHC) evaluations have shown an increase in the number of different inflammatory cells in the colon of IBS patients. In this study, we have compared the number of intraepithelial lymphocytes (IELs), eosinophils, mast cells and CD3+ T cells, in IBS patients and normal subjects. Materials and Methods: In 2016, seventy-nine patients with IBS and seventy-nine healthy subjects who underwent colonoscopy for other non-specific causes and with no pathologic findings, were enrolled in this cross-sectional study. Biopsy specimens obtained from the colon were stained, using IHC methods to determine the number of IELs, eosinophils, mast cells and CD3+ T cells. Quantitative and qualitative variables were compared between the two groups, using a Chi-square test and Student’s t-test. Results: Seventy-nine patients with IBS, 79.7% females with a mean age of 42.5±14.6 years, were recruited, as the case group, and seventy-nine individuals, 51.9% females with a mean age of 39.7±18.9 years, were enrolled as controls. The average number of IELs per high power fields (hpf) was found to be higher in the IBS group, and this difference was statistically significant (32.8±11.8 vs. 28.6±12.9; P=0.034). Also, the mean count/hpf of CD3+ T lymphocytes (23.1±7.9 vs. 20.2±8.1; P=0.024) and mast cells (7.6±3.1 vs. 6.6±3.0; P=0.041) were significantly higher in the IBS group, compared to the control group. The number of eosinophils was higher in the IBS group, but the differences were not statistically significant (P=0.066). Conclusion: According to the results, we suggest that analysis of immune cells and IELs in intestinal biopsies might be an appropriate method for diagnosis of IBS. [GMJ.2018;7:e1063]

Quantitation of Colonic Cells as Severity Markers in Patients with Irritable Bowel Syndrome.

Background: Irritable bowel syndrome (IBS) is the most common gastrointestinal syndrome. Routine histopathology and immunohistochemistry (IHC) evaluations have shown an increase in the number of different inflammatory cells in the colon of IBS patients. In this study, we have compared the number of intraepithelial lymphocytes (IELs), eosinophils, mast cells and CD3+ T cells, in IBS patients and normal subjects. Materials and Methods: In 2016, seventynine patients with IBS and seventy-nine healthy subjects who underwent colonoscopy for other non-specific causes and with no pathologic findings, were enrolled in this cross-sectional study. Biopsy specimens obtained from the colon were stained, using IHC methods to determine the number of IELs, eosinophils, mast cells and CD3+ T cells. Quantitative and qualitative variables were compared between the two groups, using a Chi-square test and Student’s t-test. Results: Seventy-nine patients with IBS, 79.7% females with a mean age of 42.5±14.6 years, were recruited, as the case group, and seventy-nine individuals, 51.9% females with a mean age of 39.7±18.9 years, were enrolled as controls. The average number of IELs per high power fields (hpf) was found to be higher in the IBS group, and this difference was statistically significant (32.8±11.8 vs. 28.6±12.9; P=0.034). Also, the mean count/hpf of CD3+ T lymphocytes (23.1±7.9 vs. 20.2±8.1; P=0.024) and mast cells (7.6±3.1 vs. 6.6±3.0; P=0.041) were significantly higher in the IBS group, compared to the control group. The number of eosinophils was higher in the IBS group, but the differences were not statistically significant (P=0.066). Conclusion: According to the results, we suggest that analysis of immune cells and IELs in intestinal biopsies might be an appropriate method for diagnosis of IBS.

Lymphocytic esophagitis: an update on histologic diagnosis, endoscopic findings, and natural history.

Lymphocytic esophagitis is a histologic pattern of injury characterized by increased intraepithelial lymphocytes (>20/high-power field) with rare, or absent granulocytes. Lymphocytes tend to be more numerous in the peripapillary epithelium, and are often associated with evidence of mucosal injury, edema, and scattered dyskeratotic cells. More than a decade following its original description, lymphocytic esophagitis remains an enigmatic entity with variable clinical presentations, associated disorders, etiologies, treatment, and natural history. Most of the confusion regarding the clinical significance of this disorder stems from its diagnostic criteria: lymphocytic esophagitis is currently defined based entirely on histologic criteria, despite the common occurrence of lymphocytosis in a variety of unrelated inflammatory conditions of the esophagus. The goal of this review is to summarize the literature regarding lymphocytic esophagitis and focus on key clinicopathologic features that distinguish it from other esophageal disorders that can show increased numbers of intraepithelial lymphocytes.

Obesity and Intraepithelial Lymphocyte Dysfunction

Abstract Obesity has become prevalent in the United States with over 30% of adults and 17% of children having a BMI over 30. Complications associated with obesity include chronic nonhealing wounds in the skin and increased susceptibility to more severe colitis in the intestine suggesting that obesity leads to barrier dysfunction. Intraepithelial lymphocytes regulate epithelial homeostasis through the production of growth factors and cytokines. Our laboratory has shown that obese mice exhibit reduced numbers of intraepithelial lymphocytes in both the skin and the intestine. The remaining intraepithelial lymphocytes have reduced growth factor and cytokine production resulting in dysregulation of the barrier. To better understand the mechanism of T cell dysfunction, we placed mice on high-fat diet immediately post-weaning when intraepithelial lymphocytes are seeding the intestine. After 7 weeks, mice fed normal chow had normal numbers of intraepithelial lymphocytes seeding the intestine, however mice fed a high-fat diet had almost 50% fewer cells. CCR9 and CD103 were both reduced on the lymphocytes suggesting problems with homing and retention. In addition, mice administered a high-fat diet were more susceptible to DSS-induced colitis. To determine whether intestinal epithelial cells could be restored to normal numbers in obese mice, obese mice were administered normal chow for 7 weeks. Indeed, reducing fat intake was sufficient to restore intraepithelial lymphocytes to normal levels and rescue mice from more severe colitis suggesting that dieting is effective in improving intestinal barrier function in obesity.

Comparative study of effects of assemblages AII and BIV of Giardia duodenalis on mucosa and microbiota of the small intestine in mice

AimsGiardiasis is one of the major causes of diarrhea worldwide and its symptoms vary in intensity, which can be attributed to different parasite assemblages. The goal of the present study was to compare the effects of infection caused by assemblages AII and BIV ofGiardia duodenalis on the response of the small intestine, microbiota, and behavioral parameters in mice. Main methodsSwiss mice were infected with assemblages AII and BIV of G. duodenalis for 15 days. Leucometry, pain, intestinal microbiota and histological parameters of the duodenum and jejunum were evaluated in the experimental groups. Key findingsBoth assemblages modified the composition of the intestinal microbiota. Infection with assemblage AII promoted leukocytosis, reflected by increasing number of polymorphonuclear cells, intraepithelial lymphocytes and pain-related behavior, indicating that this was the more aggressive assemblage with regard to its effects on the intestinal mucosa and duodenum. SignificanceThe specific assemblage of the parasite is an important parameter that affects symptomatology in the host.

The interplay of BMP4 and IL‑7 regulates the apoptosis of intestinal intraepithelial lymphocytes under conditions of ischemia̸reperfusion.

The number and function of intestinal intraepithelial lymphocytes (IELs) have been found to be significantly reduced following induction of acute intestinal mucosal damage via intestinal ischemia/reperfusion (I/R). However, the mechanism underlying this reduction remains unclear. Therefore, it was hypothesized that the interplay of bone morphogenetic protein (BMP)4 and interleukin (IL)-7 regulates IEL function and number. Recent studies have demonstrated that the different components of the BMP2/4 signaling pathway are expressed in intestinal epithelial cells (IECs) via the activation of nuclear factor (NF)-κB. In the present study, reverse transcription-polymerase chain reaction analysis and flow cytometry demonstrated that IELs express BMP receptors (BMPRIA, BMPRIB, ActRIA and BMPRII) and non-canonical signal transduction molecules (NF-κB). An in vivo mouse intestinal I/R model was used, and I/R was shown to increase the expression of BMP4 in IECs and upregulate the expression levels of BMPRIA, BMPRIB and phosphorylated NF-κB in IELs. Following isolation and culture of IELs, it was observed that exogenous BMP4 also upregulated the expression of BMPRIA and BMPRIB and activated NF-κB signaling in IELs, inducing IEL apoptosis. In addition, the rate of apoptosis of IELs decreased following treatment with the BMP-specific antagonist NOGGIN or with the NF-κB inhibitor pyrrolidine dithiocarbamate. Furthermore, it was observed that exogenous IL-7 can decrease BMP4 protein expression in IECs and the expression of phosphorylated NF-κB protein in IELs. The findings of the present study suggest that, under conditions of I/R, IEC-derived BMP4 activates NF-κB signaling in IELs, inducing IEL apoptosis, further aggravating the dysfunction of the intestinal mucosal barrier. However, these effects may be alleviated by IL-7 treatment. Therefore, BMP4 and IL-7 appear to be involved in the interaction between IECs and IELs and in the mechanism underlying intestinal mucosal barrier dysfunction.

Edwardsiella tarda outbreak affecting fishes and aquatic birds in Brazil

Background: Edwardsiella tarda infections are frequent causes of severe outbreaks in the fish farming industry besides representing possible zoonotic risks. However, naturally occurring outbreaks that affect various species besides fishes are seldom described.Aim: To report an outbreak of acute mortality caused by E. tarda affecting multiple species that inhabited a natural pond in the state of São Paulo, Brazil.Materials and methods: Three adult tilapias, three Mallard ducks and one Snow egret were necropsied and subjected to further microbiological tests. Gross and microscopic lesions were documented. The antibiotic susceptibility and phylogenetic similarities among fish and avian strains were also determined. The E. tarda species was confirmed through MALDI-TOF, partial sodB sequencing and phylogenetic analysis.Results: Macroscopical findings between the three species included intestinal dilatation, mucosal hyperaemia and mucous to liquid contents. Common histopathology findings included acute enteritis, increased number of intraepithelial lymphocytes with bacteria adhered to the intestinal epithelium and lymphoid depletion in the spleen. E. tarda was isolated from several organs from all affected species. The phylogeny employing amplified fragment length polymorphism (AFLP) of eleven strains revealed high similarity (>90%) among the isolates regardless of the affected species or sampled organs. Ten isolates of E. tarda showed susceptibility to all tested antibiotics.Conclusions: E. tarda was identified as the cause of death of the species examined. Further studies would be necessary to determine the virulence of these strains and the possible risks regarding public health.

Role of biopsy in diagnosis and treatment of adult celiac disease.

Celiac disease (CD) is an immune-mediated enteropathy that characteristically responds to treatment with a gluten-free diet. In most, clinical features improve with resolution of diarrhea and weight loss. Serological studies also tend to normalize. Small intestinal biopsies from the duodenum reveal a severe to moderately severe architectural disturbance showing crypt epithelial hyperplasia with increased numbers of epithelial cell mitotic figures along with villous "flattening", increased numbers of lamina propria plasma cells and lymphocytes and increased numbers of intra-epithelial lymphocytes in untreated disease. With a gluten-free diet, these changes can be expected to resolve to normal. In some patients, this mucosal inflammatory process may persist, especially in the proximal small intestine for variable periods of time. In CD, resolution of histopathological changes can occur within 6 months, but often, more than a year is required, and sometimes, 2 years or more. Changes are not only time-dependent, but appear to be gender-dependent with resolution more readily achieved in females compared to males, and age-dependent with more persistence of the inflammatory process in the elderly compared to younger patients. Future studies need to take into account the individual nature of the normal mucosal healing process in CD treated with a gluten-free diet.

Toxoplasma gondii infection causes structural changes in the jejunum of rats infected with different inoculum doses

AimTo evaluate the mucosal tunic and submucosal plexus of the jejunum of rats infected with different inoculum doses of Toxoplasma gondii. Main methodsRats were infected with different inoculum doses (50, 500, 1000 and 5000 oocysts) of the T. gondii for 30days, while a control group (CG) received saline solution. Blood and feces were collected before euthanasia for analysis of blood and fecal leukocytes (LEs). Histological analysis of the mucosa, submucosa, villi, crypts and enterocytes were performed. Goblet cells, intraepithelial lymphocytes (IELs) and Paneth cells were quantified. Immunohistochemistry was used to assess enteroendocrine serotonergic (5HT-IR) cells, proliferative cells (PCNA+) and mast cells. Whole mounts were obtained to determine the total submucosal neurons by Giemsa staining and metabolically active neurons (NADH-d+), nitrergic neurons (NADPH-d+) and glial cells (S100). Key findingsAn increase in blood LEs was observed 30days post-infection (dpi). Fecal LEs were more abundant in the feces in all infected groups at 21 dpi when compared to the CG. The number of IELs, sulfomucin-producing goblet cells, Paneth cells, PCNA+ cells and mast cells increased, whereas the number of 5HT-IR cells decreased. The jejunal architecture was altered, with atrophy of the mucosa, submucosa, villi and crypts. The number of total submucosal neurons decreased, but the NADPH-d+ subpopulation increased. SignificanceThe results show how chronic toxoplasmic infection affects the tissue and cellular composition of the rat jejunum. These structural changes tend to intensify with the inoculum dose, demonstrating the importance of the parasitic load on intestinal alterations.

Flow cytometry of duodenal intraepithelial lymphocytes improves diagnosis of celiac disease in difficult cases

Diagnosis of celiac disease is difficult when the combined results of serology and histology are inconclusive. Studies using flow cytometry of intraepithelial lymphocytes (IELs) have found that celiac patients have increased numbers of γδ IELs, along with a decrease in CD3-CD103 + IELs. The objective of this article is to assess the role of flow cytometric analysis of IELs in the diagnosis of celiac disease in difficult cases. A total of 312 patients with suspicion of celiac disease were included in the study. Duodenal biopsy samples were used for histological assessment and for flow cytometric analysis of IELs. In 46 out of 312 cases (14.7%) the combination of serology and histology did not allow the confirmation or exclusion of celiac disease. HLA typing had been performed in 42 of these difficult cases. Taking into account HLA typing and the response to a gluten-free diet, celiac disease was excluded in 30 of these cases and confirmed in the remaining 12. Flow cytometric analysis of IELs allowed a correct diagnosis in 39 out of 42 difficult cases (92.8%) and had a sensitivity of 91.7% (95% CI: 61.5% to 99.8%) and a specificity of 93.3% (95% CI: 77.9% to 99.2%) for the diagnosis of celiac disease in this setting. Flow cytometric analysis of IELs is useful for the diagnosis of celiac disease in difficult cases.

Pathological Changes in Gut and Liver Due to Dietary Inclusion of Betaine and Chromium Pico Linate in Nile Tilapia Fish

1. Abstract A complementary study was conducted to investigate histomorphometric changes in gut and liver due to dietary supplementation of betaine and chromium pico linate (Cr- Pic) or blend of them in Nile tilapia. Fish were divided into four groups; first group was fed a Cr- Pic or betaine free basal diet, other three groups were fed on basal diet supplemented with 800 μ g/kg Cr-Pic, 10 g/kg of betaine and betaine+Cr-Pic (10 g/kg of betaine and 800 μ g Cr-Pic). Morphometric measurements made in the intestine included counting numbers of Goblet Cells (GCs), Eosinophilic Granulocytes (EGs) and Intra Epithelial Lymphocytes (IEL), villous height, height of the enterocytes (EH) and thickness of muscular coat. Histomorphometric analysis showed that betaine increased number of GCs and IEL. Cr-Pic increased number of GCs, EGs and <span style=font-size:10.0pt; font-family: times= new= roman,serif;mso-bidi-language:ar-eg=> thickness of muscular coat. Betaine+Cr-Pic increased number of EGs, length of EH, villous height, perimeter and area , thickness of muscular coat and decreased Hepatocytes/Nuclear (H/N) ratio . We concluded that the dietary inclusion of both betaine and Cr-Pic was beneficial for liver and gut health in Nile tilapia. 2. Keywords: Betaine; Chrome; Fish; Liver; Pathology

Cytotoxic tumour-infiltrating T lymphocytes influence outcome in resected pancreatic ductal adenocarcinoma

BackgroundWe studied the prognostic effect of CD3-, CD8- and CD103-positive T lymphocytes in a cohort of 165 patients with resected pancreatic ductal adenocarcinomas (PDACs) of the treatment group (adjuvant gemcitabine) and the untreated control group of the CONKO-001 study. MethodsImmunohistochemical stainings on tissue microarrays (TMAs) against CD3, CD8 and CD103 were performed according to standard procedures. ResultsA high number of CD8-positive lymphocytes were significantly and independently associated with longer disease-free survival (DFS) and overall survival (OS) in the overall study population. Median DFS/OS were 7.4/18.1 months for patients with a low number of CD8-positive intratumoural lymphocytes (≤42 per 1 mm tissue core) and 12.7/25.2 months for patients with high numbers (>42 per 1-mm tissue core; p = 0.008/0.020; HR 0.62/0.65). The ratio of intraepithelial to total CD103-positive lymphocytes, but not total numbers of CD103-positive lymphocytes or CD103-positive intraepithelial lymphocytes, was associated with significantly improved DFS and OS in the overall study population (p = 0.022/0.009). Median DFS/OS was 5.9/15.7 for patients with a ratio of intraepithelial to total CD103-positive intratumoural lymphocytes higher than 0.3 and 11.6/24.7 for patients with a lower ratio. ConclusionT-lymphocyte subpopulations might be prognostic in resectable PDAC but need standardization and verification by further studies.

Probiotics and Probiotic Metabolic Product Improved Intestinal Function and Ameliorated LPS-Induced Injury in Rats.

In the present study, we sought to determine the effects of Bacillus subtilis (BAS) and Bacillus licheniformis (BAL) in rats after lipopolysaccharide (LPS)-induced acute intestinal inflammation. We also determined whether the B. subtilis metabolic product (BASM) is as effective as the live-cell probiotic. 60 male SD rats were randomly assigned to five groups and administered a diet containing 0.05% B. licheniformis (BAL group), 0.05% B. subtilis (BAS group), 0.5% B. subtilis metabolic product (BASM group), or a basic diet (PC group and NC group) for 40days. On day 40, BAL, BAS, BASM, and NC groups were injected with 4mg/kg body weight LPS. 4h later, all rats were anesthetized and sacrificed. The results showed that the administration of B. licheniformis and B. subtilis improved intestinal function as evidenced by histology, increased enzyme activity, and mucosal thickness. They also increased the number of intraepithelial lymphocytes and decreased mucosal myeloperoxidase activity and plasma TNF-α. In addition, the cecal content of B. subtilis-treated rats had significantly increased microbial diversity, decreased numbers of Firmicutes, and increased numbers of Bacteroidetes as compared to rats fed basic diets. Similar to BAS group, the cecal content of B. licheniformis-treated rats decreased the number of Firmicutes. Administration of B. subtilis metabolic product had similar effects on intestinal function, inflammation response, and microbial diversity as B. subtilis but these effects were attenuated. In conclusion, administration of probiotic strains B. licheniformis or B. subtilis improved intestinal function, ameliorated the inflammation response, and modulated microflora after LPS-induced acute inflammation in rats. Non-living cells also exerted probiotic properties but live cells tended to function better.