Quantification of liver steatosis is clinically relevant in various liver diseases but cannot be done by conventional sonography, which only provides a qualitative assessment with significant ...observer variability. The aim of this study was to assess sonography as an objective tool for the quantification of liver steatosis.
Files of 111 patients with chronic liver disease who were referred for sonographically guided liver biopsy were collected. A hepatorenal sonographic index was calculated on the basis of the ratio between the echogenicity of the liver and that of the right kidney cortex using histogram echo intensity. Liver steatosis was graded by histology.
A significant correlation was found between histologic steatosis and the hepatorenal sonographic index (r = 0.82, p < 0.001). The validity of the hepatorenal sonographic index for the diagnosis of fatty liver was compared with liver biopsies with a steatosis level > 5%. The area under the receiver operating characteristic curve was 99.2% (95% CI, 98-100%). The optimal hepatorenal sonographic index cutoff point for the prediction of steatosis > 5% was 1.49, with sensitivity of 100% and specificity of 91%. The optimal hepatorenal sonographic index cutoff point for the prediction of steatosis >/= 25% was 1.86, with sensitivity of 90% and specificity of 90%. The optimal hepatorenal sonographic index cutoff point for the prediction of steatosis >/= 60% was 2.23, with sensitivity of 90% and specificity of 93%.
The hepatorenal sonographic index is a sensitive noninvasive method for steatosis quantification. It can diagnose small amounts of liver fat that would be missed by conventional sonography. It is reproducible and operator independent and can serve as an efficient tool to follow patients with steatosis and evaluate the efficacy of new treatment techniques.
Intestinal epithelial cells (IECs) are the first to encounter luminal antigens and may be involved in intestinal immune responses. Fungi are important components of the intestinal microflora. The ...potential role of fungi, and in particular their cell wall component β‐glucan, in modulating human intestinal epithelial responses is still unclear. Here we examined whether human IECs are capable of recognizing and responding to β‐glucans, and the potential mechanisms of their activation. We show that human IECs freshly isolated from surgical specimens, and the human IEC lines HT‐29 and SW480, express the β‐glucan receptor Dectin‐1. The β‐glucan‐consisting glycans curdlan and zymosan stimulated IL‐8 and CCL2 secretion by IEC lines. This was significantly inhibited by a Dectin‐1 blockade using its soluble antagonist laminarin. Spleen tyrosine kinase (Syk), a signaling mediator of Dectin‐1 activation, is expressed in human IECs. β‐glucans and Candida albicans induced Syk phosphorylation, and Syk inhibition significantly decreased β‐glucan‐induced chemokine secretion from IECs. Thus, IECs may respond to β‐glucans by the secretion of pro‐inflammatory chemokines in a Dectin‐1‐ and Syk‐dependent pathway, via receptors and a signaling pathway described to date only for myeloid cells. These findings highlight the importance of fungi–IEC interactions in intestinal inflammation.
Background: The effect of 5-fluorouracil (5-FU) on activated lymphocytes was explored. Materials and Methods: The in vitro
effects of 5-FU on DNA synthesis in mitogen-activated lymphocytes from ...healthy volunteers were compared to those of the antimetabolites
doxorubicin, cyclophosphamide and 6-mercaptopurine. These effects were assessed by alterations in the phenotypic profile and
the percentage of cells in various phases of the cell cycle, as well as by the secretion of T helper (Th)1 and Th2 cytokines
(ELISA). Results: Unlike 5-FU, the other antimetabolites failed to augment DNA synthesis in activated lymphocytes. The effect
of 5-FU correlated with an increase in the percentage of cells in the S-phase and caused an increased in CD4 + cells, a decrease in CD56 + cells and a shift of the cytokine secretion pattern from Th2 to Th1. Conclusion: 5-FU exhibited a unique effect on DNA synthesis
in activated lymphocytes which was accompanied by selective effects on various lymphocyte subpopulations.
Background & Aims
Primary biliary cholangitis (PBC) is a progressive‐cholestatic autoimmune liver disease. Dendritic cells (DC) are professional antigen‐presenting cells and their prominent presence ...around damaged bile ducts of PBC patients are documented. cDC1 is a rare subset of DC known for its cross‐presentation abilities and interleukin 12 production. Our aim was to assess the role of cDC1 in the pathogenesis of PBC.
Methods
We utilized an inducible murine model of PBC and took advantage of the DC reporter mice Zbtb46gfp and the Batf3−/− mice that specifically lack the cDC1 subset. cDC1 cells were sorted from blood of PBC patients and healthy individuals and subjected to Bulk‐MARS‐seq transcriptome analysis.
Results
Histopathology assessment demonstrated peri‐portal inflammation in wild type (WT) mice, whereas only minor abnormalities were observed in Batf3−/− mice. Flow cytometry analysis revealed a two‐fold reduction in hepatic CD8/CD4 T cells ratio in Batf3−/− mice, suggesting reduced intrahepatic CD8 T cells expansion. Histological evidence of portal fibrosis was detected only in the WT but not in Batf3−/− mice. This finding was supported by decreased expression levels of pro‐fibrotic genes in the livers of Batf3−/− mice. Transcriptome analysis of human cDC1, revealed 78 differentially expressed genes between PBC patients and controls. Genes related to antigen presentation, TNF and IFN signalling and mitochondrial dysfunction were significantly increased in cDC1 isolated from PBC patients.
Conclusion
Our data illustrated the contribution the cDC1 subset in the pathogenesis of PBC and provides a novel direction for immune based cell‐specific targeted therapeutic approach in PBC.
The complement system regulator CD55 was initially found to carry the Cromer blood group system antigens, and its complete loss of function was subsequently revealed to cause a severe monogenic ...gastrointestinal syndrome characterized by protein-losing enteropathy and susceptibility to venous thrombosis. Here we present homozygosity to the
CD55
c.596C>T; p.Ser199Leu variant, which was previously described as the Cromer Dr(a−) genotype, in two Bukharan Jewish CD55-deficiency patients with variable disease severity. We confirm that this missense variant causes aberrant splicing and deletion of 44 bp in exon 5, leading to premature termination and low expression of the CD55 protein. Furthermore, Patient 1 exhibited a mildly abnormal B cell immunophenotyping profile. By population screening we established that this variant is highly prevalent in the Bukharan Jewish population, with a carrier frequency of 1:17, suggesting that many similar patients are un- or mis-diagnosed. The phenotypic variability, ranging from abdominal pain when eating a high-fat diet to the full CD55-deficiency phenotype, is likely related to modifiers affecting the proportion of the variant that is able to escape aberrant splicing. Establishing the diagnosis of CD55-deficiency in a timely manner, even in patients with milder symptoms, may have a critical effect on their management and quality-of-life since treatment with the complement inhibitor eculizumab is highly effective in ameliorating disease manifestations. Awareness of founder mutations within certain populations can further guide genetic testing and prevent a diagnostic odyssey, by placing this
CD55
variant high on the differential diagnosis.
As a large number of cancers are caused by nonsense mutations in key genes, read‐through of these mutations to restore full‐length protein expression is a potential therapeutic strategy. Mutations in ...the adenomatous polyposis coli (APC) gene initiate the majority of both sporadic and hereditary colorectal cancers (CRC) and around 30% of these mutations are nonsense mutations. Our goal was to test the feasibility and effectiveness of APC nonsense mutation read‐through as a potential chemo‐preventive therapy in Familial Adenomatous Polyposis (FAP), an inherited CRC syndrome patients. Ten FAP patients harboring APC nonsense mutations were treated with the read‐through inducing antibiotic erythromycin for 4 months. Endoscopic assessment of the adenomas was performed at baseline, after 4 and after 12 months. Adenoma burden was documented in terms of adenoma number, maximal polyp size and cumulative polyp size per procedure. Tissue samples were collected and subjected to molecular and genetic analyses. Our results show that in the majority of patients the treatment led to a decrease in cumulative adenoma burden, median reduction in cumulative adenoma size and median reduction in adenoma number. Molecular and genetic analyses of the adenomas revealed that the treatment led to a reduced number of somatic APC mutations, reduced cellular proliferation and restoration of APC tumor‐suppressing activity. Together, our findings show that induced read‐through of APC nonsense mutations leads to promising clinical results and should be further investigated to establish its therapeutic potential in FAP and sporadic CRCs harboring nonsense APC mutations.
What's new?
Enhanced translational read‐through, in which ribosomes ignore premature stop codons to generate full‐length proteins, is a promising strategy for the treatment of malignancies arising from nonsense mutations. Here, nonsense mutation read‐through of the adenomatous polyposis coli (APC) gene, via treatment with the antibiotic erythromycin, was investigated in patients with inherited colorectal cancer syndrome. Patients treated with erythromycin for four months showed reductions in adenoma burden, including decrease in adenoma size and number. Read‐through treatment was associated with decreased APC mutation percentage and restoration of APC tumor suppressor activity. The results warrant further study of erythromycin read‐through therapy as a chemopreventive approach.
Objectives:
Societies’ guidelines suggest routine tissue sampling in all children undergoing esophagogastroduodenoscopy and ileocolonoscopy, even in the absence of visible endoscopy abnormalities. We ...aimed to determine the agreement between endoscopic and histopathological findings in pediatric endoscopy and to assess the yield of routine biopsies from all sites.
Methods:
Since January 2019, our endoscopy institute protocol has included routine biopsies sampling from the esophagus, stomach, duodenum, ileum, and colon in all diagnostic procedures. Agreement between tests was done using the kappa coefficient (κ). The study included all endoscopies performed during 2019.
Results:
In total, 541 diagnostic endoscopies were done during the study period with 434 (80%) esophagogastroduodenoscopy and 107 (20%) were ileocolonoscopy. Compared to histology, endoscopic findings performance were: esophagus—sensitivity 33%, specificity 98%; stomach—sensitivity 60%, specificity 89%; duodenum—sensitivity 50%, specificity 97%; duodenal bulb—sensitivity 47%, specificity 89%; terminal ileum—sensitivity 82%, specificity 100%; colon—sensitivity 84%, specificity 96%. Assessment of concordance between endoscopic and histopathologic findings reveals an overall low level of agreement in esophagogastroduodenoscopy (κ of 0.39, 0.51, 0.53, and 0.24 for the esophagus, stomach, duodenal second part, and bulb, respectively), and good agreement in ileocolonoscopy (κ of 0.88 and 0.81 for the ileum and colon, respectively).
Conclusions:
Endoscopy findings are highly specific for histologic pathology, whereas the absence of findings correlates poorly with histologic findings. Ileocolonoscopy shows better agreement than esophagogastroduodenoscopy. Our data support routine tissue sampling in pediatric endoscopy.
Abstract only
e14634
Background: "Personalized medicine,” is the tailoring of medical treatment to a single person aiming to maximize efficacy and minimize toxicity. Currently there is no good ...prediction for response to therapy. The Chick Chorioallantoic Membrane (CAM) is naturally immuno-deficient and rich in vascularity therefore an ideal system, allowing generate 3D cancerous “organoids” in a very efficient, reproducible and cost-effective manner and translates basic research to the clinic. Aims: Generate a “personalized” HTP system for a quick, reliable and effective evaluation of different therapeutic options using 3D tumors in a "humanized egg" instead of mouse PDX model. Methods: Fertilized eggs were incubated until day 3 (37°C, 75-90% humidity). Then, 2ml of albumin was pulled out to separate the CAM from the eggshell and a small window in the eggshell has been made. To destroy the chicken immune system development, the eggs were irradiated at day 5 and human immune cells were then inoculated onto the CAM. On day 7, single cells suspension or tissues, derived from cancer patients, were transplanted onto the CAM and visible tumors were performed ("CAM-PDX"). Drugs, mAbs and chemotherapy, were applied via the yolk sac. Tumor growth was measured, weighted and monitored by caliper and IVIS fluorescent imaging platform. IHC was performed and confirmed the response of the particular specimens to the tested regiment. Results: Histology and IHC analysis confirmed that the established tumors retained their characteristics. Positive Ki-67 staining confirmed that cancer cells proliferate while the treated tumors showed reduced staining. Anti-CD24 mAb, FOLFOX, cetuximab, Foflorinox and Gemcitabine, given as single agent or combinations, successfully inhibited CR and pancreatic tumors (by 70-75%). Detection of active caspase 3 confirmed those results. Biopsies from human specimens, were successfully established and expanded by serial passages allows generation of bio-bank. The stimulated human PBMCs demonstrated enhanced proliferation in vitro and in ovo, even after 5 days in the egg. Irradiated eggs showed no functional immune system even after 2 weeks of development. Conclusions: The CAM is an ideal, effective, economical and powerful avatar-based precision medicine approach to predict the best protocol for cancer therapy.
To evaluate molecular profiles in the small bowel (SB) mucosa proximal to the pouch in ulcerative colitis (UC) patients after pouch surgery.
Patients were prospectively recruited and stratified ...according to disease behaviour: normal pouch (NP), chronic pouchitis (CP), and Crohn's-like disease of the pouch (CLDP). Biopsies obtained from the pouch and the normal-appearing proximal SB (40 cm proximal to the anal verge) were compared to ileal biopsies from normal controls (NC). A histopathological score based on the degree of polymorphonuclear and mononuclear infiltrates was used to assess inflammation in the pouch and the proximal SB. Gene expression analysis was performed using microarrays, and validated by real-time PCR. Gene ontology and clustering were evaluated by bioinformatics.
Thirty-six subjects were recruited (age 18-71 years, 16 males). Histopathology scores demonstrated minimal differences in the normal-appearing proximal SB of all groups. Nonetheless, significant (fold change ≥2, corrected p FDR ≤ 0.05) molecular alterations in the proximal SB were detected in all groups (NP n=9; CP n=80; and CLDP n=230) compared with NC. The magnitude of DUOX2 alteration in the proximal SB was highest. An increase of 6.0, 9.8 and 21.7 folds in DUOX2 expression in NP, CP, CLDP, respectively was observed. This was followed by alterations in MMP1, SLC6A14 and PGC. Gene alterations in the proximal SB overlapped with alterations within the pouch (76% and 97% overlap in CP and CLDP, respectively). Gene ontology analysis in the proximal SB and pouch were comparable.
Significant gene expression alterations exist in an apparently unaffected proximal SB. Alterations in the pouch and the proximal SB were comparable, suggesting that inflammation may not be limited to the pouch, but that it extends to the proximal SB.
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