Chromogranin A-positive Cells Research Articles

291-LB: Possible Alpha-Cell Regeneration from Mucin-Producing Ducts in Zinc-Binding Site-Mutated CRY1-Expressing Diabetic Mice

C414A-CRY1 transgenic mice (TG) show the symptoms of diabetes due to cellular senescence-like phenotypes in their β-cells as we have reported at ADA Scientific Sessions in 2015. Due to the cellular phenotypes, structural remodeling in islets of TG progresses with age (S. Okano et al., ADA 2016-2022): the increase of the population ratios of α-cell to β-cell and of δ-cell to β-cell as well as the occurrence of fibrillation, angiogenesis and also the generation of mucin-producing intra-islet ductal cells (IIDCs) were observed. Contrary to normal ductal cells, IIDCs only weakly react with DBA lectin, indicating unique characters different from normal ductal cells (ADA 2019). Also, we have shown the results suggesting that δ-cells may play some role as the source for β-cells to supply new β-cells in TG (ADA 2022). To examine the possible roles of IIDCs to maintain of the amount of pancreatic endocrine cells under hyperglycemia for prolonged period in TG, in this study, we conducted detailed immune-staining experiments which focused on fully developed IIDCs, not small ones that still not fully developed. We distinguished IIDCs from normal ducts, PanINs as well as PDGs as the measure of the reactivity to DBA-lectin. Chromogranin A-positive cells were observed in IIDCs, but not in normal ducts, PanINs and PDGs. Then we examined IIDCs with anti-hormone antibodies. In the region of developed IIDCs, cytokeratin 17/19 and glucagon double-positive cells were predominantly observed. Only small portion of somatostatin-positive cells were clearly located adjacent to IIDCs. Cluster of β-cells were located apart from IIDCs. In addition, scrutinized observations suggested the delamination of glucagon-positive cells from the ductal epithelium of the IIDCs. Taken together, these results strongly suggest the possible neogenesis of endocrine cells in IIDCs, that is, IIDCs and their lining cells play roles as the source for endocrine cells, particularly for α, and also for δ cells. Disclosure S. Okano: None. A. Yasui: None. S. Kanno: None. Y. Sasaki: None. K. Satoh: None. M. Igarashi: None. O. Nakajima: None. Funding Grants-in-Aid for Scientific Research (19K07498); Tohoku University Institute of Development, Aging and Cancer

PS-R03-4: TRANSARTERIAL EMBOLIZATION, THORACIC SPINE DECOMPRESSION AND FUSION OF METASTATIC PHEOCHROMOCYTOMA: A CASE REPORT

Surveillance of metastatic lesions and biochemical control of catecholamines in patients with malignant pheochromocytoma remain a fundamental problem to be solved. Here we present a 60-year-old male who visited a local clinic because of headache, sweating, malaise, and loss of body weight. Hypertension, orthostatic hypotension, along with remarkable increase in serum and urine noradrenaline, and urine metanephrine and vanillylmandelic acid were noted, and pheochromocytoma was suspected accordingly. Magnetic resonance imaging failed to reveal adrenal gland or sympathetic plexuses, yet it uncovered metastatic lesions in the liver and spine. Functional imaging study using 131I-Metaiodobenzylguanidine (MIBG) scan detected notable uptake in the spinal lesion, albeit not in the hepatic lesion. On the other hand, positron emission tomography with fluorodeoxyglucose (FDG-PET) disclosed FDG uptake both in the hepatic and spinal masses. Biopsy of the hepatic tumors revealed chromogranin A-positive clusters of cells, a finding consistent with metastatic pheochromocytoma. Transarterial embolization (TAE) of the metastatic tumors in the liver was performed for the purpose of tumor mass reduction. Additionally, TAE followed by resection of the spinal lesion was conducted. Pathological examination of the spinal tumor also showed chromogranin A-positive cells, leading to diagnosis of malignant pheochromocytoma. Crucially, TAE of the liver and spinal tumors resulted in dramatic reduction of catecholamine production, likely constituting an effective therapeutic strategy against metastatic pheochromocytoma (Homma, et al. Hypertens Res 2006). Afterwards, the patient became unable to walk because of weakness in the left leg at the age of 64. It seemed attributed to spinal cord compression from the tumor at the level of thoracic vertebrae 7. The patient underwent posterior decompression and fusion, and his muscle strength of the left leg was recovered. At the age of 65, the patient developed paralytic ileus following a surgery against metastatic tumors in the skull. In this case report, we discuss early detection of metastatic lesions and biochemical control of catecholamines in patients with malignant pheochromocytoma.

Purification of human iPSC-derived cells at large scale using microRNA switch and magnetic-activated cell sorting.

SummaryFor regenerative cell therapies using pluripotent stem cell (PSC)-derived cells, large quantities of purified cells are required. Magnetic-activated cell sorting (MACS) is a powerful approach to collect target antigen-positive cells; however, it remains a challenge to purify various cell types efficiently at large scale without using antibodies specific to the desired cell type. Here we develop a technology that combines microRNA (miRNA)-responsive mRNA switch (miR-switch) with MACS (miR-switch-MACS) to purify large amounts of PSC-derived cells rapidly and effectively. We designed miR-switches that detect specific miRNAs expressed in target cells and controlled the translation of a CD4-coding transgene as a selection marker for MACS. For the large-scale purification of induced PSC-derived cardiomyocytes (iPSC-CMs), we transferred miR-208a-CD4 switch-MACS and obtained purified iPSC-CMs efficiently. Moreover, miR-375-CD4 switch-MACS highly purified pancreatic insulin-producing cells and their progenitors expressing Chromogranin A. Overall, the miR-switch-MACS method can efficiently purify target PSC-derived cells for cell replacement therapy.

ID2 controls differentiation of enteroendocrine cells in mouse small intestine.

The mammalian gut is the largest endocrine organ. Dozens of hormones secreted by enteroendocrine cells regulate a variety of physiological functions of the gut but also of the pancreas and brain. Here, we examined the role of the helix-loop-helix transcription factor ID2 during the differentiation of intestinal stem cells along the enteroendocrine lineage. To assess the functions of ID2 in the adult mouse small intestine, we used single-cell RNA sequencing, genetically modified mice, and organoid assays. We found that in the adult intestinal epithelium Id2 is predominantly expressed in enterochromaffin and peptidergic enteroendocrine cells. Consistently, the loss of Id2 leads to the reduction of Chromogranin A-positive enteroendocrine cells. In contrast, the numbers of tuft cells are increased in Id2 mutant small intestine. Moreover, ablation of Id2 elevates the numbers of Serotonin+ enterochromaffin cells and Ghrelin+ X-cells in the posterior part of the small intestine. Finally, ID2 acts downstream of BMP signalling during the differentiation of Glucagon-like peptide-1+ L-cells and Cholecystokinin+ I-cells towards Neurotensin+ PYY+ N-cells. ID2 plays an important role in cell fate decisions in the adult small intestine. First, ID2 is essential for establishing a differentiation gradient for enterochromaffin and X-cells along the anterior-posterior axis of the gut. Next, ID2 is necessary for the differentiation of N-cells thus ensuring a differentiation gradient along the crypt-villi axis. Finally, ID2 suppresses the commitment of secretory intestinal epithelial progenitors towards tuft cell lineage and thus controls host immune response to commensal and parasitic microbiota.

Extend of neuroendocrine differentiation in adenocarcinoma prostate needle biopsies

Neuroendocrine differentiation in prostatic adenocarcinoma is now one of the emerging prognostic factor that determine treatment outcome. This study aims to identify the extent of neuroendocrine differentiation amidst the tumour cells in adenocarcinoma prostate with the help of immunohistochemical marker, Chromogranin A(CgA) and correlate with its Gleason score. A semiquantative scoring system was utilised to estimate and quantify cells having neuroendocrine differentiation.Out of 40 cases of prostatic adenocarcinomas in needle core biopsies, 93% of patients with a Gleason score >6 showed CgA positivity of more than 5% and 2cases (16.6%) having Gleason score 9 showed >50% of CgA positive cells in the tumour area.Hence higher Gleason score correlated with higher degree of neuroendocrine differentiation in our study.

(Phospho)proteomic Profiling of Microsatellite Unstable CRC Cells Reveals Alterations in Nuclear Signaling and Cholesterol Metabolism Caused by Frameshift Mutation of NMD Regulator UPF3A.

DNA mismatch repair-deficient colorectal cancers (CRCs) accumulate numerous frameshift mutations at repetitive sequences recognized as microsatellite instability (MSI). When coding mononucleotide repeats (cMNRs) are affected, tumors accumulate frameshift mutations and premature termination codons (PTC) potentially leading to truncated proteins. Nonsense-mediated RNA decay (NMD) can degrade PTC-containing transcripts and protect from such faulty proteins. As it also regulates normal transcripts and cellular physiology, we tested whether NMD genes themselves are targets of MSI frameshift mutations. A high frequency of cMNR frameshift mutations in the UPF3A gene was found in MSI CRC cell lines (67.7%), MSI colorectal adenomas (55%) and carcinomas (63%). In normal colonic crypts, UPF3A expression was restricted to single chromogranin A-positive cells. SILAC-based proteomic analysis of KM12 CRC cells revealed UPF3A-dependent down-regulation of several enzymes involved in cholesterol biosynthesis. Furthermore, reconstituted UPF3A expression caused alterations of 85 phosphosites in 52 phosphoproteins. Most of them (38/52, 73%) reside in nuclear phosphoproteins involved in regulation of gene expression and RNA splicing. Since UPF3A mutations can modulate the (phospho)proteomic signature and expression of enzymes involved in cholesterol metabolism in CRC cells, UPF3A may influence other processes than NMD and loss of UPF3A expression might provide a growth advantage to MSI CRC cells.

Chronic restraint stress induces serotonin transporter expression in the rat adrenal glands

Chronic restraint stress (CRS) magnifies restraint-induced corticosterone secretion through a mechanism involving increased adrenocortical 5-HT content and turnover. We analysed the impact of CRS on serotonin transporter (SERT) expression and distribution in rat adrenal glands. Male Wistar rats were submitted to CRS (20 min/day) or undisturbed control conditions for 14 days. Exposure to CRS induced a remarkable increase in SERT-like immunoreactivity in the adrenal cortex, which closely matched that of chromogranin A immunostaining, along with a significant increase in SERT protein and mRNA levels in whole adrenals as determined by immunohistochemistry, Western blot and RT-PCR assays, respectively; all these CRS-induced changes occurred almost exclusively in left adrenals. Closely similar results were obtained in animals that received a 14-day chronic corticosterone treatment. These results unravel an interesting association between chronic stress exposure and SERT expression in adrenocortical chromogranin A-positive cells, which seems to be a glucocorticoid-dependent phenomenon.

Neuroendocrine differentiation in usual-type prostatic adenocarcinoma: Molecular characterization and clinical significance.

Small cell neuroendocrine (NE) carcinomas of the prostate classically lose androgen receptor (AR) expression, may harbor loss of the RB1, TP53, and PTEN tumor suppressor genes, and are associated with a poor prognosis. However usual-type adenocarcinomas may also contain areas of NE differentiation, and in this context the molecular features and biological significance are less certain. We examined the molecular phenotype and oncologic outcomes of primary prostate adenocarcinomas with ≥5% NE differentiation (≥5% chromogranin A-positive NE cells in any given tumor spot on tissue microarray) using threeindependent study sets: a set of tumors with paneth cell-like NE differentiation (n = 26), a retrospective case-cohort of intermediate- and high-risk patients enriched for adverse outcomes (n = 267), and primary tumors from a retrospective series of men with eventual castration-resistant metastatic prostate cancer (CRPC) treated with abiraterone or enzalutamide (n = 55). Benign NE cells expressed significantly lower quantified AR levels compared withpaired benign luminal cells (P < .001). Similarly, paneth-like NE carcinoma cells or carcinoma cells expressing chromogranin A expressed significantly lower quantified AR levels than paired non-NE carcinoma cells (P < .001). Quantified ERG protein expression, was also lower in chromogranin A-labeled adenocarcinoma cells compared withunlabeled cells (P < .001) and tumors with NE differentiation showed lower gene expression scores for AR activity compared withthose without. Despite evidence of lower AR signaling, adenocarcinomas with NE differentiation did not differ by prevalence of TP53 missense mutations, or PTEN or RB1 loss, compared withthose without NE differentiation. Finally, NE differentiation was not associated with time to metastasis in intermediate- and high-risk patients (P = .6 on multivariate analysis), nor with progression-free survival in patients with CRPC treated with abiraterone or enzalutamide (P = .9). NE differentiation in usual-type primary prostate adenocarcinoma is a molecularly and clinically distinct form of lineage plasticity from that occurring in small cell NE carcinoma.

Human norovirus targets enteroendocrine epithelial cells in the small intestine

Human noroviruses are a major cause of diarrheal illness, but pathogenesis is poorly understood. Here, we investigate the cellular tropism of norovirus in specimens from four immunocompromised patients. Abundant norovirus antigen and RNA are detected throughout the small intestinal tract in jejunal and ileal tissue from one pediatric intestinal transplant recipient with severe gastroenteritis. Negative-sense viral RNA, a marker of active viral replication, is found predominantly in intestinal epithelial cells, with chromogranin A-positive enteroendocrine cells (EECs) identified as a permissive cell type in this patient. These findings are consistent with the detection of norovirus-positive EECs in the other three immunocompromised patients. Investigation of the signaling pathways induced in EECs that mediate communication between the gut and brain may clarify mechanisms of pathogenesis and lead to the development of in vitro model systems in which to evaluate norovirus vaccines and treatment.

Increased Chromogranin A-Positive Hormone-Negative Cells in Chronic Pancreatitis.

Chronic pancreatitis (CP) is characterized by inflammation, fibrosis, and a loss of pancreatic acinar cells, which can result in exocrine and eventually endocrine deficiency. Pancreatitis has been reported to induce formation of new endocrine cells (neogenesis) in mice. Our recent data have implicated chromogranin A-positive hormone-negative (CPHN) cells as potential evidence of neogenesis in humans. We sought to establish if CPHN cells were more abundant in CP in humans. We investigated the frequency and distribution of CPHN cells and the expression of the chemokine C-X-C motif ligand 10 (CXCL10) and its receptor chemokine C-X-C motif receptor 3 in pancreas of nondiabetic subjects with CP. CPHN cell frequency in islets was increased sevenfold in CP [2.1% ± 0.67% vs 0.35% ± 0.09% CPHN cells in islets, CP vs nonpancreatitis (NP), P < 0.01], as were the CPHN cells found as scattered cells in the exocrine areas (17.4 ± 2.9 vs 4.2 ± 0.6, CP vs NP, P < 0.001). Polyhormonal endocrine cells were also increased in CP (2.7 ± 1.2 vs 0.1 ± 0.04, CP vs NP, % of polyhormonal cells of total endocrine cells, P < 0.01), as was expression of CXCL10 in α and β cells. There is increased islet endogenous expression of the inflammation marker CXCL10 in islets in the setting of nondiabetic CP and an increase in polyhormonal (insulin-glucagon expressing) cells. The increase in CPHN cells in CP, often in a lobular distribution, may indicate foci of attempted endocrine cell regeneration.

Interleukin-17C in Human Helicobacter pylori Gastritis.

The interleukin-17 (IL-17) family of cytokines (IL-17A to IL-17F) is involved in many inflammatory diseases. Although IL-17A is recognized as being involved in the pathophysiology of Helicobacter pylori-associated diseases, the role of other IL-17 cytokine family members remains unclear. Microarray analysis of IL-17 family cytokines was performed in H. pylori-infected and uninfected gastric biopsy specimens. IL-17C mRNA was upregulated approximately 4.5-fold in H. pylori-infected gastric biopsy specimens. This was confirmed by quantitative reverse transcriptase PCR in infected and uninfected gastric mucosa obtained from Bhutan and from the Dominican Republic. Immunohistochemical analysis showed that IL-17C expression in H. pylori-infected gastric biopsy specimens was predominantly localized to epithelial and chromogranin A-positive endocrine cells. IL-17C mRNA levels were also significantly greater among cagA-positive than cagA-negative H. pylori infections (P = 0.012). In vitro studies confirmed an increase in IL-17C mRNA and protein levels in cells infected with cagA-positive infections compared to cells infected with either cagA-negative or cag pathogenicity island (PAI) mutant. Chemical inhibition of IκB kinase (IKK), mitogen-activated protein extracellular signal-regulated kinase (MEK), and Jun N-terminal kinase (JNK) inhibited induction of IL-17C proteins in infected cells, whereas p38 inhibition had no effect on IL-17C protein secretion. In conclusion, H. pylori infection was associated with a significant increase in IL-17C expression in human gastric mucosa. The role of IL-17C in the pathogenesis of H. pylori-induced diseases remains to be determined.

Benign intestinal glandular lesions in the vagina: a possible correlation with implantation.

BackgroundEnteric-type glandular lesions are extremely rare in the vagina. Their histological origin remains a matter of speculation at present.MethodWe review two rectal mucosal prolapse-like polyps and one intestinal-type adenosis in the vagina.ResultsCase 1, a 64-year-old woman, presented with a vaginal polypoid lesion with a size of 4 × 3 × 3 cm. Case 2, an 8-year-old girl, had a 1.5 × 1.5 × 0.8-cm pedunculated polyp in the vaginal navicular fossa and a clinically suspected rectovaginal fistula. Case 1 and 3 had an obsolete severe perineal laceration. On histopathological examination, cases 1 and 2 resembled rectal mucosal prolapse or inflammatory cloacogenic polyp (rectal mucosal prolapse-like polyp). Case 3 had an incidental intestinal-type adenosis in the removed vaginal wall. Immunohistochemistry confirmed the intestinal differentiation in all 3 lesions by showing diffuse CDX2-positive, CK20-positive, and scattered chromogranin A-positive neuroendocrinal cells in the lower compartment of the crypt.ConclusionsIn summary, we report herein three unusual cases of benign intestinal-type glandular lesions in the vagina including two rectal mucosal prolapse-like polyps and one case of intestinal-type adenosis, and discuss possibilities for their histogenetic basis.

Brachyury identifies a class of enteroendocrine cells in normal human intestinal crypts and colorectal cancer.

Normal homeostasis of adult intestinal epithelium and repair following tissue damage is maintained by a balance of stem and differentiated cells, many of which are still only poorly characterised. Enteroendocrine cells of the gut are a small population of differentiated, secretory cells that are critical for integrating nutrient sensing with metabolic responses, dispersed amongst other epithelial cells. Recent evidence suggests that sub-sets of secretory enteroendocrine cells can act as reserve stem cells. Given the link between cells with stem-like properties and cancer, it is important that we identify factors that might provide a bridge between the two. Here, we identify a sub-set of chromogranin A-positive enteroendocrine cells that are positive for the developmental and cancer-associated transcription factor Brachyury in normal human small intestinal and colonic crypts. Whilst chromogranin A-positive enteroendocrine cells are also Brachyury-positive in colorectal tumours, expression of Brachyury becomes more diffuse in these samples, suggesting a more widespread function in cancer. The finding of the developmental transcription factor Brachyury in normal adult human intestinal crypts may extend the functional complexity of enteroendocrine cells and serves as a platform for assessment of the molecular processes of intestinal homeostasis that underpins our understanding of human health, cancer and aging.

Establishment of Leptin-Responsive Cell Lines from Adult Mouse Hypothalamus.

Leptin resistance is considered to be the primary cause of obesity. However, the cause of leptin resistance remains incompletely understood, and there is currently no cure for the leptin-resistant state. In order to identify novel drug-target molecules that could overcome leptin resistance, it would be useful to develop in vitro assay systems for evaluating leptin resistance. In this study, we established immortalized adult mouse hypothalamus—derived cell lines, termed adult mouse hypothalamus (AMH) cells, by developing transgenic mice in which SV40 Tag was overexpressed in chromogranin A—positive cells in a tamoxifen-dependent manner. In order to obtain leptin-responsive clones, we selected clones based on the phosphorylation levels of STAT3 induced by leptin. The selected clones were fairly responsive to leptin in terms of STAT3, ERK, and Akt phosphorylation and induction of c-Fos mRNA induction. Pretreatment with leptin, insulin, and palmitate attenuated the c-Fos mRNA response to leptin, suggesting that certain aspects of leptin resistance might be reconstituted in this cellular model. These cell lines are useful tools for understanding the molecular nature of the signal disturbance in the leptin-resistant state and for identifying potential target molecules for drugs that relieve leptin resistance, although they have drawbacks including de-differentiated nature and lack of long-time stability.

Tu1712 Analysis of the IL-17C-IL-17RE Cytokine Ligand-Receptor Pair in Human Inflammatory Bowel Disease Reveals Chromogranin a-Positive Epithelial Cells As Major Producers of IL-17C and IL-17RE-Positive Lamina Propria Mononuclear Cells As Main Colonic IL-17C Targets

Background: To date, the role of IL-17C and its specific receptor subunit IL-17RE in inflammatory bowel disease (IBD) settings is entirely based on murine models. Therefore, the aim of this study was to analyze the role of the novel IL-17 family member IL-17C and its receptor IL-17RE in human IBD. Methods: IL-17C expression in intestinal epithelial cells (IEC) was analyzed by qPCR, immunoblotting, immunohistochemistry, EMSA, RNA interference and pharmacological or antibody-mediated inhibition. IL-17C protein and mRNA expression in IBD patient samples was assessed by ELISA, qPCR and immunohistochemistry. IL-17C serum levels were correlated to IL23R genotype status. Results: IL-17A amplifies IL-17C expression in human IEC upon TNF-α stimulation; this depends on TNFα-activated NF-κB, ERK-1/2 and p38 MAP kinases, and IL-17A-activated Akt kinase, MCPIP and C/EBP-δ pathways. IL-17C serum levels are elevated in patients with ulcerative colitis (UC) (P=0.008). IL-17C mRNA is increased in inflamed colonic sections of IBD patients (P=0.005) and correlates significantly with the mRNA expression of Th1 cytokines (e.g., IFN-γ; P<0.01) and Th17 cytokines (e.g., IL-17A, IL-23, CCL20; P<0.01). Genotyping revealed that UC patients carrying a minor allele of the IL23R SNP rs134351 have decreased IL-17C serum levels (P=0.037). Immunohistochemistry displayed increased IL-17C and IL17RE staining in IBD patients with active disease, compared to IBD patients in remission and healthy controls. Chromogranin A-positive IEC are major producers of IL-17C in the colon, whereas the IL-17RE subunit was only present on lamina propria mononuclear cells but not in IEC. In vitro experiments suggested that IEC lines are not responsive to IL-17C stimulation, even when combined with other cytokines, such as IL-22. Conclusions: This is the first comprehensive study involving human IBD patients which demonstrates an upregulation of the novel IL-17C-IL-17RE cytokine ligand-receptor pair during active human IBD. In contrast to murine models, IL-17RE expression is confined to lamina propria leukocytes and not epithelial cells; specialized chromogranin A-positive cells are among the major producers of IL-17C in the epithelium. Our data indicate that serum IL-17C is modulated by IL23R genotype status. In addition, our data reveal potential novel targets for the pharmacological modulation of epithelium-derived IL-17C.

Tu1713 Role of miR-19b-Targeting SOCS3 in Modulating Production of Chemokine in Colonic Epithelial Cells and Is Associated With the Pathogenesis of Crohn's Disease

Background: To date, the role of IL-17C and its specific receptor subunit IL-17RE in inflammatory bowel disease (IBD) settings is entirely based on murine models. Therefore, the aim of this study was to analyze the role of the novel IL-17 family member IL-17C and its receptor IL-17RE in human IBD. Methods: IL-17C expression in intestinal epithelial cells (IEC) was analyzed by qPCR, immunoblotting, immunohistochemistry, EMSA, RNA interference and pharmacological or antibody-mediated inhibition. IL-17C protein and mRNA expression in IBD patient samples was assessed by ELISA, qPCR and immunohistochemistry. IL-17C serum levels were correlated to IL23R genotype status. Results: IL-17A amplifies IL-17C expression in human IEC upon TNF-α stimulation; this depends on TNFα-activated NF-κB, ERK-1/2 and p38 MAP kinases, and IL-17A-activated Akt kinase, MCPIP and C/EBP-δ pathways. IL-17C serum levels are elevated in patients with ulcerative colitis (UC) (P=0.008). IL-17C mRNA is increased in inflamed colonic sections of IBD patients (P=0.005) and correlates significantly with the mRNA expression of Th1 cytokines (e.g., IFN-γ; P<0.01) and Th17 cytokines (e.g., IL-17A, IL-23, CCL20; P<0.01). Genotyping revealed that UC patients carrying a minor allele of the IL23R SNP rs134351 have decreased IL-17C serum levels (P=0.037). Immunohistochemistry displayed increased IL-17C and IL17RE staining in IBD patients with active disease, compared to IBD patients in remission and healthy controls. Chromogranin A-positive IEC are major producers of IL-17C in the colon, whereas the IL-17RE subunit was only present on lamina propria mononuclear cells but not in IEC. In vitro experiments suggested that IEC lines are not responsive to IL-17C stimulation, even when combined with other cytokines, such as IL-22. Conclusions: This is the first comprehensive study involving human IBD patients which demonstrates an upregulation of the novel IL-17C-IL-17RE cytokine ligand-receptor pair during active human IBD. In contrast to murine models, IL-17RE expression is confined to lamina propria leukocytes and not epithelial cells; specialized chromogranin A-positive cells are among the major producers of IL-17C in the epithelium. Our data indicate that serum IL-17C is modulated by IL23R genotype status. In addition, our data reveal potential novel targets for the pharmacological modulation of epithelium-derived IL-17C.

Distribution of chromogranin A in human fetal pancreas.

Immunohistochemical study of distribution of chromogranin A (neuroendocrine cell marker) in the developing human pancreas was carried out. Immunopositive reaction to chromogranin A was detected in the primordial pancreas from the early period of development (8 weeks). The count of chromogranin A-positive cells in early fetuses significantly surpassed that of insulin- and glucagon-containing cells. Chromogranin A immunoreactive cells, but not cells reacting with antibodies to insulin and glucagon were detected in the pancreatic islets and pancreatic ducts during all periods of gestation. These results suggested that hormone synthesis by pancreatic endocrine cells was preceded by the expression of chromogranin A and hence, this marker could be used for studies of the mechanisms of development of the endocrine part of the pancreas.

The effect of ileal interposition surgery on enteroendocrine cell numbers in the UC Davis type 2 diabetes mellitus rat

AimTo investigate the short-term effect of ileal interposition (IT) surgery on gut morphology and enteroendocrine cell numbers in the pre-diabetic UC Davis type 2 diabetes mellitus (UCD-T2DM) rat. Study designTwo-month old male UCD-T2DM rats underwent either sham (n=5) or IT (n=5) surgery. Intestines were collected 1.5months after surgery. The jejunum, ileum and colon regions were processed for histochemical and immunohistochemical labeling and stereological analyses of changes in gut morphometry and number of enteroendocrine cells. ResultsStereological analysis showed that intestinal volume, luminal surface area and the number of all chromogranin A-positive enteroendocrine cells were markedly increased in the IT rats compared with sham-operated animals. Subanalyses of the glucagon-like peptide 2, cholecystokinin, serotonin cells and the neurotensin immunoreactive sub-pool of enteroendocrine cells in the IT region revealed an increase in numbers across phenotypes. However, the density of the different cell types varied. ConclusionIT surgery in the UCD-T2DM rat leads to rapid alterations in gut morphometry and an increase in the number of enteroendocrine cells. This effect may potentially explain why IT surgery delays the onset of type 2 diabetes in the UCD-T2DM rat.

Chromogranin a staining as a prognostic variable in newly diagnosed Gleason score 7-10 prostate cancer treated with definitive radiotherapy

To demonstrate the association of neuroendocrine differentiation, as identified by chromogranin A (CgA) staining, with clinical outcomes in newly diagnosed prostatic adenocarcinoma treated with definitive radiotherapy (RT). Patients with Gleason score ≥7 adenocarcinoma were identified from our outcomes database. RT consisted of external beam, brachytherapy, or external beam with brachytherapy boost. Biopsy specimens were stained for neuroendocrine differentiation with CgA. Results were interpreted by a single pathologist. CgA staining was quantified as 0%, <1%, 1-10%, or >10% of tumor cells. Clinical outcomes were blinded at the time of pathologic evaluation. CgA staining was performed on 289 patients. 149 patients had Gleason score 7, and 140 were Gleason score 8-10. Median follow-up was 6.5 years. For patients with <1% versus >1% CgA staining, pretreatment characteristics were well-balanced. CgA staining was detected in 90 cases (31%). 58 patients had focal positive (<1%) CgA staining, and 32 cases had >1% of tumor cells CgA positive. Patients with >1% CgA staining had inferior biochemical control, clinical failure, distant metastases (DM), and cause-specific survival (CSS) rates. Ten-year rates of DM were 8% versus 48% for patients with <1% versus >1% CgA positive cells, respectively (P < 0.001). CSS at 10 years was 95% versus 76%, respectively (P < 0.001). Local control was equivalent in the two patient cohorts. Patients with <1% CgA staining had similar outcomes to those patients with 0% staining. Neuroendocrine differentiation involving >1% of tumor cells on prostate cancer biopsies is a predictor of DM and CSS in patients treated with primary RT.

Distinctive Histopathologic Findings of Pancreatic Hamartomas Suggesting Their “Hamartomatous” Nature

Pancreatic hamartoma is a rare tumor, and its characteristic histopathologic features have not yet been fully evaluated. In this study, we collected 9 cases of pancreatic hamartoma to elucidate distinctive histopathologic features that can serve to establish this tumor as a clear disease entity and thus formulate useful histopathologic criteria for this tumor. The cases comprised 4 men and 5 women with a mean age of 62.7 years. The average tumor diameter was 3.3 cm. All patients underwent surgical treatment, and none showed any recurrence postoperatively. Macroscopically, pancreatic hamartomas were well-demarcated tumors with a solid or solid and cystic appearance. Microscopically, these tumors comprised mature acini and small-sized to medium-sized ducts showing a distorted architecture with various amounts of fibrous stroma. Strikingly, the tumors consistently lacked concentric elastic fibers in their duct walls, peripheral nerves, and well-formed islets of Langerhans, all of which exist in both the normal and atrophic pancreas. Immunohistochemically, scattered chromogranin A-positive neuroendocrine cells were observed in the acinar and ductal components. Ductal components were positive for S-100 protein. Spindle-shaped stromal cells expressed CD34 and/or c-kit. These histopathologic features were distinct from those of 5 cases of pancreatic ductal adenocarcinoma, 3 cases of type 1 autoimmune pancreatitis (lymphoplasmacytic sclerosing pancreatitis), 3 cases of alcoholic chronic pancreatitis, and 5 cases of normal pancreas. In conclusion, pancreatic hamartomas share some distinctive histopathologic features and clinical outcomes (neither recurrence nor metastasis) that allow them to be interpreted as malformative lesions. The term "hamartoma" is appropriate for these unique lesions.