Chromogranin Research Articles

In-depth analysis of proteomic and genomic fluctuations during the time course of human embryonic stem cells directed differentiation into beta cells.

Pluripotent stem cells (PSC) endocrine differentiation at a large scale allows sampling of transcriptome and proteome with phosphoproteome (proteoform) at specific time points. We describe the dynamic time course of changes in cells undergoing directed beta-cell differentiation and show target proteins or previously unknown phosphorylation of critical proteins in pancreas development, NKX6-1, and Chromogranin A (CHGA). We describe fluctuations in the correlation between gene expression, protein abundance, and phosphorylation, following differentiation protocol perturbations at all stages to identify proteoform profiles. Our modeling recognizes outliers on a phenomic landscape of endocrine differentiation, and we describe new biological pathways involved. We have validated our proteomic data by analyzing independent single-cell RNAseq datasets for in-vitro pancreatic islet production and corroborated our findings for several proteins suggestive as targets for future research. The single-cell analysis combined with proteoform data places new protein targets within the specific time point and at the specific pancreatic lineage of differentiating stem cells. We suggest that non-correlating proteins abundances or new phosphorylation motifs of NKX6.1 and CHGA point to new signaling pathways that may play an essential role in beta-cell development. We present our findings for the research community's use to improve endocrine differentiation protocols and developmental studies.

Role of chromogranin A-derived fragments after resection of nonfunctioning pancreatic neuroendocrine tumors.

No single reliable biomarker is available for nonfunctioning pancreatic neuroendocrine tumors (NF-PanNETs). Vasostatin-1 (VS-1), the N-terminal fragment of chromogranin A (CgA), seems to be a more accurate biomarker compared to its precursor. Primary aim was to investigate the ability of VS-1, compared to total-CgA, to assess the effectiveness of surgical resection performed for NF-PanNETs. Secondary aim was to evaluate two additional CgA-derived fragments, pancreastatin (PST) and vasostatin-2 (VS-2), as possible biomarkers for NF-PanNETs. Consecutive patients who underwent surgery for NF-PanNETs at San Raffaele Scientific Institute were included (n = 35). Plasma levels of CgA and CgA-derived fragments were measured by Enzyme-Linked ImmunoSorbent Assay (ELISA), preoperatively and postoperatively. Preoperative VS-1 was significantly higher compared to VS-1 measured on postoperative day 5 (POD5) (pre: 0.338nM versus POD5: 0.147nM, P < 0.001), whereas total-CgA significantly increased after surgery (pre: 1.123nM versus POD5: 1.949nM, P = 0.006). Overall, 24 patients showed ≥ 1 feature of tumor aggressiveness (T3-T4, nodal/distant metastases, Ki67 > 5%, microvascular/perineural invasion, necrosis). The median percentage decrease in VS-1 plasma levels was 63% (IQR 28-88%) among patients with aggressive tumors, compared to 13% (IQR 0-57%) in the remaining population (P = 0.033). No significant differences in terms of PST (P = 0.870) and VS-2 (P = 0.909) were observed between preoperative and postoperative time. VS-1 provides an early assessment of surgical efficacy in patients who undergo resection for NF-PanNETs, especially in those with aggressive neoplasms. Total-CgA, PST and VS-2 have no clinical utility in this setting.

Catestatin induces glycogenesis by stimulating the phosphoinositide 3-kinase-AKT pathway.

Defects in hepatic glycogen synthesis contribute to post-prandial hyperglycaemia in type 2 diabetic patients. Chromogranin A (CgA) peptide Catestatin (CST: hCgA352-372 ) improves glucose tolerance in insulin-resistant mice. Here, we seek to determine whether CST induces hepatic glycogen synthesis. We determined liver glycogen, glucose-6-phosphate (G6P), uridine diphosphate glucose (UDPG) and glycogen synthase (GYS2) activities; plasma insulin, glucagon, noradrenaline and adrenaline levels in wild-type (WT) as well as in CST knockout (CST-KO) mice; glycogen synthesis and glycogenolysis in primary hepatocytes. We also analysed phosphorylation signals of insulin receptor (IR), insulin receptor substrate-1 (IRS-1), phosphatidylinositol-dependent kinase-1 (PDK-1), GYS2, glycogen synthase kinase-3β (GSK-3β), AKT (a kinase in AKR mouse that produces Thymoma)/PKB (protein kinase B) and mammalian/mechanistic target of rapamycin (mTOR) by immunoblotting. CST stimulated glycogen accumulation in fed and fasted liver and in primary hepatocytes. CST reduced plasma noradrenaline and adrenaline levels. CST also directly stimulated glycogenesis and inhibited noradrenaline and adrenaline-induced glycogenolysis in hepatocytes. In addition, CST elevated the levels of UDPG and increased GYS2 activity. CST-KO mice had decreased liver glycogen that was restored by treatment with CST, reinforcing the crucial role of CST in hepatic glycogenesis. CST improved insulin signals downstream of IR and IRS-1 by enhancing phospho-AKT signals through the stimulation of PDK-1 and mTORC2 (mTOR Complex 2, rapamycin-insensitive complex) activities. CST directly promotes the glycogenic pathway by (a) reducing glucose production, (b) increasing glycogen synthesis from UDPG, (c) reducing glycogenolysis and (d) enhancing downstream insulin signalling.

Prognostic stratification for patients with neuroendocrine tumours receiving 177Lu-Dotatate.

177Lu-Dotatate is increasingly used in patients with advanced neuroendocrine tumour (NET). However, few prognostic markers are available to stratify progression-free survival (PFS) of patients who received 177Lu-Dotatate. Clinicopathological data including baseline circulating biomarkers of patients with advanced NET who received 177Lu-Dotatate were routinely collected and were retrospectively analysed. Continuous variables were normalized by dividing them by their upper normal limits. The whole data set was randomly divided into a training set and a validation set. Univariate and multivariate logistic regression analyses were used to identify independent markers and to develop a scoring model to predict treatment failure at 1 year. In total, 195 patients were included. Elevated baseline chromogranin A (CgA), normal creatinine and previous chemotherapy were three risk factors independently associated with 1-year treatment failure. By combining these risk factors, a scoring model was developed which could accurately predict 1-year treatment failure both in the training set (area under curve, AUC, 0.813; 95% CI, 0.731-0.895; P< 0.001) and in the validation set (AUC, 0.816; 95% CI, 0.644-0.968; P< 0.001). After selecting a score of 29.7 as the cut-off value of the scoring model, patients could be stratified into two groups namely low-risk and high-risk with significantly different 1-year treatment failure rate, PFS and overall survival (OS; P< 0.001) both in the training set and validation set. In conclusion, baseline CgA, creatinine level and previous chemotherapy were independently associated with 1-year treatment failure of patients with advanced NET who received 177Lu-Dotatate and the scoring model and prognostic stratification based on these markers could accurately predict 1-year treatment failure, PFS and OS.

Serum chromogranin A correlated with albuminuria in diabetic patients and is associated with early diabetic nephropathy

BackgroundThe kidney is the main site for the removal of chromogranin A (CgA). Previous studies have found that patients with renal impairment displayed elevated concentrations of CgA in plasma and that CgA concentrations reflect a deterioration of renal function. In this study, we aimed to estimate serum CgA levels and to evaluate the role of serum CgA in the early diagnosis of diabetic nephropathy (DN).MethodsA total of 219 patients with type 2 diabetes mellitus (T2DM) were included in this cross-sectional study. These patients were classified into normoalbuminuria (n = 121), microalbuminuria (n = 73), or macroalbuminuria (n = 25) groups based on their urine albumin to creatinine ratios (UACRs). The degree of DN is reflected by UACR. A control group consisted of 45 healthy subjects. The serum CgA levels were measured by ELISA, and other key parameters were assayed.ResultsSerum CgA levels were higher in patients with T2DM than in control subjects, and a statistically significant difference among the studied subgroups regarding CgA was found (P < 0.05). The levels of serum CgA increased gradually with the degree of DN (P < 0.001). Serum CgA levels showed a moderate-intensity positive correlation with UACRs (P < 0.001). A cutoff level of 3.46 ng/ml CgA showed 69.86% sensitivity and 66.12% specificity to detect DN in the early stage.ConclusionThe levels of serum CgA increased gradually with the degree of DN and can be used as a biomarker in the early detection of DN.

Growth factors and chromogranin A in intra-abdominal adhesions of children

Pathogenesis of adhesions is still unclear and does not give a clear response whether the morphopathogenetic mechanisms of congenital and acquired adhesions in children are different or not. Thus, we searched for the appearance and distribution of growth factors, their receptors and chromogranins in adhesions of children in the ontogenetic aspect. Tissues were obtained from 17 children of group 1 (1 day to 3 years old) and 10 children of group 2 (10 to 16 years old). Visibly intact peritoneum was obtained from teenagers of group 2 and used as control tissue. Routine staining and immunohistochemistry for insulin-like growth factor 1 (IGF1), insulin-like growth factor 1 receptor (IGF IR), transforming growth factor beta (TGF-β), hepatocyte growth factor (HGF), chromogranin A (CgA) was applied. Statistical analysis included semi-quantitative evaluation of immunopositive structures, Wilcoxon signed-rank test and Wilcoxon rank sum test for comparison of data. The results showed patchy fibrosis with round-shaped fibroblasts, neoangiogenesis and inflammation. Adhesions in group 1 demonstrated various appearance of IGF, IGF IR, few to moderate number of TGF-β-, HGF- and moderate number of CgA-containing cells. In both groups, fibroblasts predominated among actively expressing cells. Group 2 showed moderate to numerous IGFI and HGF positive cells, while IGF IR- and TGF-β-containing cells were numerous. Intact peritoneum showed variable TGF-β, CgA, and few to moderate HGF, IGFI, IGF IR cells. In conclusion, an increase in IGFI, IGF IR, TGF-β and HGF characterises the acquired (“aged”) adhesions. Fibroblasts are most active in adhesion formation with the increase of cellular activity during the aging of children and adhesions. Fibroblasts with changes in shape stimulate further development of disease via intensive CgA expression.

Associations between chronic work stress and plasma chromogranin A/catestatin among healthy workers.

ObjectivesPlasma chromogranin A (CgA) may play a critical role on linking work stress to health outcomes. The aim of our study was to investigate the associations between work stress and plasma CgA levels in healthy workers without chronic diseases.MethodsThe study included 260 healthy workers from EHOP study. Work stressors were assessed by the Chinese version of the 23‐item ERI‐Q questionnaire. Plasma CgA and catestatin levels were measured by ELISA kits. The demographic characteristics were collected from medical records.ResultsAmong the final 260 subjects including 173 males (66.5%) and 87 females (33.5%), the average age was 37.6 ± 10.6 years old. Effort, overcommitment, and ERI were positively associated with plasma CgA level, respectively (r = 0.267, 0.319, and 0.304, all p < .001), while reward was negatively associated with CgA level (r = −0.237, p < .001). The workers with high effort, overcommitment, or ERI had significantly higher plasma CgA levels, while the workers with high rewards had significantly lower plasma CgA levels. The workers with both high overcommitment and high ERI had highest plasma CgA levels. In the linear regression analysis, after adjustment for confounders, effort, overcommitment, and ERI were respectively positively related to plasma CgA, while reward negatively related to plasma CgA. The associations between work stress and plasma catestatin was not significant. The ratio of CgA and catestatin was associated with work stress.ConclusionsWork stress is associated with plasma CgA which may be play a crucial role on the pathway from chronic work stress to cardiovascular diseases.

Neuroendocrine tumors (NETs) - experience of a single Center.

Introduction: Neuroendocrine neoplasms (NENs) are a heterogeneous group of tumors arising from cells that are part of the diffuse neuroendocrine system. Patients, Materials and Methods: We conducted a retrospective study in which we included a number of 91 cases diagnosed with neuroendocrine tumors (NETs). Descriptive statistics was performed: number of cases based on location, distribution by gender (male/female), distribution by age, and we also performed a morphological and immunohistochemical (IHC) study. Results: The highest number of cases was found in lungs (60 cases). Tumors located on the skin, breast or bladder have been discovered, locations considered rare for this type of tumor. Of all cases diagnosed in the lungs, 59 were diagnosed as small cell carcinomas (SCCs) and only one case as NET. All surgical specimens were positive for chromogranin A (CgA), with a different expression for the other immunomarkers. For the lung biopsies, the most frequently IHC staining was CgA and cluster of differentiation 56 (CD56), with an increased positivity for the latter. Conclusions: CgA remains the most sensitive immunomarker in the diagnosis of NETs. CD56 is the most widely used immunomarker for diagnosing small cell lung tumors. Positive expression of thyroid transcription factor 1 (TTF1) immunomarker does not confirm pulmonary origin of SCCs.

SOX11 is a sensitive and specific marker for pulmonary high-grade neuroendocrine tumors

BackgroundSynaptophysin (SYN), chromogranin A (CGA), CD56 and insulinoma-associated protein 1 (INSM1) are proposed neuroendocrine (NE) markers used for diagnosis of pulmonary NE tumors. These NE markers have been identified in subsets of non-NE tumors requiring differential diagnosis, thus we sought to explore new NE markers.MethodsWe evaluated the immunohistochemical expression of SOX11, a transcription factor involved in neurogenesis, in pulmonary NE tumors and large cell carcinomas (LCCs).ResultsWe found that SOX11 showed a sensitivity similar to INSM1 and CGA, and less than SYN and CD56 in small cell lung carcinomas (SCLCs) and large cell neuroendocrine carcinomas (LCNECs). While SOX11 is more specific than the other four markers for diagnosis of high-grade neuroendocrine carcinomas (HG-NECs) because 1) None of LCCs (0/63), the most challenging non-NE tumor type for differential diagnosis due to overlapped morphology with LCNECs displayed SOX11 positivity. While expression of at least one of SYN, CGA, CD56 or INSM1 was identified in approximately 60% (18/30) of LCCs. 2) SOX11 was only expressed in 1 of 37 carcinoid tumors in contrast to diffuse expression of SYN, CGA, CD56 and INSM1. In HG-NECs, we noticed that SOX11 was a good complementary marker for SCLC diagnosis as it was positive in 7 of 18 SYN−/CGA−/CD56− SCLCs and 3 of 8 SYN−/CGA−/CD56−/INSM1− SCLCs, and SOX11 positivity in 4 of 6 SYN−/CGA−/CD56− cases previously diagnosed as LCCs with NE morphology provides additional evidence of NE differentiation for reclassification into LCNECs, which was further confirmed by electromicroscopical identification of neurosecretory granules. We also found SOX11 expression cannot predict the prognosis in patients with HG-NECs.ConclusionsTherefore, SOX11 is a useful complementary transcriptional NE marker for diagnosis and differential diagnosis of SCLC and LCNEC.

The Effects of Psychological Stress, Depression and Inadequate Coping Behaviors on the Progression of Periodontal Disease

Aim: The aim of the present study was to explore the associations between periodontal diseases, psycho-neuro-immunologic variables like stress and depression by estimating salivary cortisol (CORT), salivary Chromogranin- A (CgA), and salivary C-reactive protein (CRP). Methodology: A total of 100 patients were included in the study and categorized into two groups which included 50 in control group (healthy individuals) and 50 recall periodontal patients. Psycho-neuro-immunologic variables like stress and depression were assessed through the Derogatis Stress Profile (DSP) and Centre of Epidemiologic Studies Depression scale (CES-D). Periodontal parameters like plaque index, gingival index, probing pocket depth and clinical attachment levels were recorded. Saliva samples were collected for estimation of stress markers such as cortisol, Chromogranin- A, and C-reactive protein. Results: Stress, depression and stress markers were correlated with measures of periodontal disease. In addition, oral care neglect during periods of stress and depression was associated with attachment loss and PD &gt;5 mm. Stress markers and CRP levels were also significantly higher in subjects with stress patients when compared to healthy individuals. Conclusions: The results of our study suggest that stress might be associated with periodontal disease through psychologic and behavioral mechanisms. Our result strengthens the suggested hypothesis of association between psychologic factors and markers of periodontal disease.

Performance Evaluation of the KRYPTOR Compact PLUS Analyzer-Based B.R.A.H.M.S. CgA Ⅱ KRYPTOR Assay for Chromogranin A Measurement

Numerous immunoassays have been developed to measure the levels of chromogranin A (CgA), a useful biomarker for diagnosing and monitoring generally heterogeneous neuroendocrine tumors (NETs). Here, we evaluated the imprecision and linearity of three such assays: KRYPTOR (ThermoFisher Scientific), NEOLISA (EuroDiagnostica), and CgA-RIA (CisBio), using 123 samples for each assay. The correlation coefficients between the assays were 0.932 (CgA-RIA versus NEOLISA), 0.956 (KRYPTOR versus CgA-RIA), and 0.873 (NEOLISA versus KRYPTOR). KRYPTOR showed good precision, with percent coefficients of variation less than 5% for low and high concentration quality controls. Linearity was maintained over a wide concentration range. Comparison of CgA levels from three disease entities (NETs, non-NET pancreatic tumors, and prostate cancer) and healthy controls showed that patients with NETs had significantly higher CgA levels (n = 57, mean: 1.82 ± 0.43 log ng/mL) than healthy individuals (n = 20, mean: 1.51 ± 0.23 log ng/mL; p = 0.018). No other significant differences between groups were observed. All three immunoassays showed strong correlations in measured CgA levels. Because KRYPTOR operation uses a fully automated random-access system and requires shorter incubation times and smaller sample volumes, the KRYPTOR assay may improve laboratory workflow while maintaining satisfactory analytical performance.

Application of serum markers in medullary thyroid carcinoma

Objective: To discuss the diagnostic value of calcitonin(CT), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), pro-gastrin releasing peptide (Pro-GRP) and chromogranin A (CgA) in the identification of medullary thyroid carcinoma (MTC). Methods: The CT levels in 105 cases of MTC, 50 cases of papillary thyroid carcinoma, 10 cases of thyroid follicular carcinoma, 5 cases of undifferentiated thyroid carcinoma, 50 cases of benign thyroid diseases, 30 cases of non-thyroid malignant tumors and 50 cases of healthy controls were measured from February 2017 to August 2019 at the Department of Clinical Laboratory, Cancer Hospital affliated to Fudan University. Additionally, 79 cases of MTC, 30 cases of non-MTC thyroid malignant tumors and 30 healthy controls were selected for the measurement of CEA, NSE, Pro-GRP and CgA levels. The receiver operating curve was utilized to clarify the area under the curve (AUC), sensitivity, and specificity of each indicator to distinguish between different groups. Results: The medians of CT concentrations in the group of MTC patients was 607.2 (152.5,2 777.5)pg/ml, which was statistically significantly higher than that of the subjects in the group of papillary thyroid carcinoma 1.48 (0.5,2.91)pg/ml, follicular thyroid carcinoma 1.90 (0.82,2.99)pg/ml, undifferentiated thyroid carcinoma 0.50 (0.50,4.93)pg/ml, benign thyroid disease 1.30 (0.50,2.79)pg/ml, non-thyroid malignancies 1.36 (0.50,2.89)pg/ml and healthy controls 2.05 (0.89,3.18)pg/ml. The sensitivity, specificity and AUC of CT to distinguish MTC vs. non-MTC patients was 96.2%, 99.3% and 0.99, respectively. The maximum diameter (>1 cm, P=0.001, OR=15.74) and number (>1, P=0.04, OR=3.4) of nodules were two independent risk factors for elevated CT. CEA (AUC=0.94), NSE (AUC=0.65), Pro-GRP (AUC=0.94) and CgA (AUC=0.83) could all distinguish MTC vs. non-MTC thyroid malignancies. The AUC, sensitivity and specificity by combining CT, CEA, NSE, Pro-GRP and CgA to differentiate MTC vs. non-MTC thyroid malignancies was 1, 100% and 100%, respectively. Conclusions: CT, CEA, NSE, Pro-GRP and CgA may be helpful for the auxiliary diagnosis of MTC. The combination of these indicators in the diagnosis of MTC has high sensitivity and specificity.

Neuroendocrine cells in prostate cancer correlate with poor outcomes: a systematic review and meta-analysis.

To perform a systematic review and meta-analysis of the literature to understand the variation in the reporting of neuroendocrine staining and determine the influence of reporting neuroendocrine staining at diagnosis on patient outcomes. Medical databases were searched to identify studies in which adenocarcinoma specimens were stained with any of the following four neuroendocrine markers: chromogranin A (CgA), neuron-specific enolase (NSE), synaptophysin and CD56. The prevalence of neuroendocrine staining and correlation of the prevalence of neuroendocrine staining to patient outcomes were analysed using a random-effects model. All statistical tests were two-sided. Sixty-two studies spanning 7616 patients were analysed. The pooled prevalence for the most common marker, CgA (41%), was similar to that of NSE (39%) and higher than that of synaptophysin (31%). The prevalence of CgA staining was significantly influenced by reporting criteria, where objective thresholds reduced the variation in prevalence to 26%. No correlation was found between CgA prevalence and tumour grade. Patients positive for CgA staining using objective criteria had more rapid biochemical progression (hazard ratio [HR] 1.98, 95% confidence interval [CI] 1.49 to 2.65) and poorer prostate cancer-specific survival (HR 7.03, 95% CI 2.55 to 19.39) compared to negative patients, even among those with low-risk cancers. Discrepancies in the reported prevalence of neuroendocrine cells in adenocarcinoma are driven by the inconsistent scoring criteria. This study unequivocally demonstrates that when neuroendocrine cell staining is assessed with objective criteria it identifies patients with poor clinical outcomes. Future studies are needed to determine the exact quantifiable thresholds for use in reporting neuroendocrine cell staining to identify patients at higher risk of progression.

Characterization of novel neuropeptide proteolytic processing involved in dementia pathophysiology

AbstractBackgroundNeuropeptides are neurotransmitter‐like molecules with multiple roles in neuronal activity that are processed into functional peptides (aka proteoforms) by prohormone convertases and other currently unidentified proteases. Both neuropeptides and their proteoforms are implicated in the pathogenesis of different forms of dementia. The neuropeptide VGF is decreased in the cerebrospinal fluid (CSF) and brain of patients with Alzheimer’s disease (AD) and other forms of dementia. VGF356‐375 and VGF1‐40 decrease in the CSF of patients with AD and frontotemporal dementia (FTD), respectively, compared to healthy age‐matched controls. Although many such neuropeptide proteoforms are thought to be involved in dementia, current research has focused on the neuropeptides themselves, leaving the proteoforms and their physiological processing pathways understudied and their contribution to dementia pathogenesis unclear.MethodUsing recombinant proteolysis assays, we aimed to investigate the proteolytic processing of several neuropeptides involved in dementia. Neuropeptides of interest were identified by mass spectrometry analysis of pooled CSF samples obtained from five AD patients and five healthy controls. Proteasix, the MEROPS, and the human protein atlas databases were used to identify novel neuronally‐expressed proteases that may cleave these identified neuropeptides; proteases of interest were selected based on the predicted probability of cleavage. Recombinant protein assays and immunoblots with neuropeptide‐specific antibodies were used to investigate the ability of these proteases to cleave the neuropeptides, and banding patterns were confirmed in triplicate.ResultTwenty‐two neuronally expressed proteases previously linked to dementia were identified that may cleave the neuropeptides chromogranin A (CHGA), secretogranin‐1 (SCG1), secretogranin‐2 (SCG2), secretogranin‐3 (SCG3), and VGF. The proteases a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4), calpain‐1 (CAPN1), cathepsin S (CTSS), granzyme A (GZMA), and matrix metalloprotease‐3 (MMP‐3) displayed high probability for neuropeptide proteolysis. These proteases cleaved the neuropeptides into N‐terminal, mid‐region, or C‐terminal neuropeptide proteoforms, and all neuropeptide‐protease combinations except the SCG3‐CAPN1 pair demonstrated cleavage.ConclusionOverall, this work provides mechanistic insight into neuropeptide processing and expands our understanding of neuropeptide proteoform involvement in dementia. Future projects aim to confirm and expand upon these results using mass‐spectrometry, cell culture systems, and CSF and brain protein analysis.

Welfare and performance of sows and piglets in farrowing pens with temporary crating system on a Spanish commercial farm

The study aimed to compare the welfare and performance of sows and piglets in three different farrowing systems (farrowing crate (FC) and farrowing pen with temporary crating (TC): SWAP and JLF15). One batch of crossbred Duroc was followed in every season. There were 18 sows (183 piglets) in FC, 23 sows (243 piglets) in SWAP, and 23 sows (237 piglets) in JLF15. Farrowing day was Day (D) 0 and weaning occurred on D24. Crating period was from entry to weaning in FC, and from 1-day pre-expected farrowing day to D3 in TC. Social interactions between littermates, between sows and piglets, and exploration by sows and piglets were observed on D2, D4, D12, and D23. Video recordings of crushing events which led to piglets’ death were studied. Piglets were weighed on D3 and D19, and foreleg abrasions were assessed on D19. Sow saliva samples were collected on 1 day before and after temporary confinement, D2, D4, D12, and D23 to evaluate cortisol (CORT) and chromogranin A (CgA) levels. TC piglets initiated naso-naso contacts with sows more frequently than FC on D2 (P ≤ 0.01). TC sows interacted with piglets more frequently than FC on D12 (P ≤ 0.05). TC sows explored the pens more frequently than FC on D4 (P ≤ 0.05). Crushing rate (i.e. number of piglets per sow) of SWAP (1.2 ± 0.3) was higher than those of JLF15 (0.6 ± 0.2) and FC (0.3 ± 0.1) (P ≤ 0.02), and crushing rate of JLF15 was higher than that of FC (P < 0.0001). Autumn batch showed the lowest crushing rate amongst different batches (P < 0.001). Number of crushing incidents was similar when TC sows were crated or loose (P = 0.54). The percentage of occasions when sows used a support from the pen when changing posture but still crushed the piglets was higher in FC than in TC (P = 0.05). No difference of growth and foreleg abrasion in piglets were found between systems. CORT in SWAP peaked on D2 (P = 0.02), and CgA in JLF15 peaked on D4 and remained elevated on D12 (P ≤ 0.05). CORT and CgA in FC remained similar during lactation. Our results suggested that TC facilitated mother-young interactions; TC showed a higher crushing rate; sows changed their posture differently between systems and batches; the practice of temporary crating did not alter the level of stress biomarkers in sows.

INSM1, a Novel Biomarker for Detection of Neuroendocrine Neoplasms: Cytopathologists' View.

Background: Insulinoma-associated protein 1 (INSM1) has been considered as a novel immunostain for neuroendocrine tumors (NETs) and is hypothesized to be more reliable than first-generation NET biomarkers, such as CGA (chromogranin A), SYP (synaptophysin) and CD56 (neural cell adhesion molecule). In this review, we summarize existing literature on INSM1′s reliability as an immunostain for detection of various NETs, its results in comparison to first-generation NET biomarkers, and its expression in both non-NETs and benign tissues/cells on cytology specimens (cell blocks/smears).

The Prognostic and Predictive Role of Chromogranin A in Gastroenteropancreatic Neuroendocrine Tumors - A Single-Center Experience.

Chromogranin A (CgA) is a non-specific biomarker excreted by neuroendocrine tumor (NET) cells. Elevation of circulating CgA level can be detected in gastroenteropancreatic (GEP)-NET patients and has been shown to correlate with tumor burden. The prognostic and predictive roles of CgA level and the change of CgA level are controversial. In this study, we retrospectively analyzed 102 grade 1/2 GEP-NET patients with available baseline or serial follow-up CgA levels from the National Cheng Kung University Hospital to evaluate the association between circulating CgA level and the tumor extent, overall survival (OS), and tumor response prediction. The baseline characteristics, baseline CgA level, and change of CgA level during follow-up and their association was analyzed. Sixty cases had baseline CgA levels available prior to any treatment and ninety-four cases had serial follow-up CgA levels available during treatment or surveillance. Baseline CgA levels were associated with stage and sex. Higher baseline CgA levels were associated with worse OS after adjusting for sex, stage, grade, primary site, and functionality (hazard ratio=13.52, 95% confidence interval (CI), 1.06-172.47, P=0.045). The cross-sectional analysis for the change of CgA level during follow-up showed that a ≥ 40% increase of CgA meant a higher probability of developing tumor progression or recurrence than those with a < 40% increase of CgA level (odds ratio=5.04, 95% CI, 1.31-19.4, P=0.019) after adjusting for sex, age, grade, stage, and functionality. Our study results suggest that CgA may be a predictive marker for tumor burden, OS, and tumor progression in GEP-NET patients.

Focal neuroendocrine carcinoma mixed with adenocarcinoma of the gallbladder with aggressive lymph node metastasis in a patient who did not meet the mixed neuroendocrine\u2013non-neuroendocrine neoplasm criteria

A 70-year-old Japanese woman who was treated for interstitial pneumonia (IP) with steroid therapy developed cholecystitis. A serial computed-tomography (CT) imaging showed irregular thickness of the fundus wall of the gallbladder and two rapidly enlarged lymph nodes (LNs): number (no.) 12 and no. 8a. Positron-emission tomography-computed tomography (PET-CT) scan showed an abnormal uptake at the site of the gallbladder tumor and those LNs. We subsequently performed open radical cholecystectomy and LN dissection of the no. 12 and 8a LNs, following complete remission of IP. The histology showed gallbladder adenocarcinoma, with a single focus of neuroendocrine carcinoma (NEC) component of less than 30%; Ki-67 index > 80%, synaptophysin (Syn) (+), chromogranin A (CgA) (+), and clusters of differentiation (CD) 56 (+) (T2bN1M0, Stage IIIB). LN no. 8a was diffusely metastatic with NEC components. LN no. 12c, which was adjacent to the cystic duct, revealed necrosis without apparent tumor cells, but was highly suspicious for tumor necrosis. The final diagnosis was adenocarcinoma of the gallbladder with focal NEC (< 30%), which did not meet the criteria for mixed neuroendocrine–non-neuroendocrine neoplasm (MiNEN). Postoperatively, she completed 4 cycles of adjuvant chemotherapy for NEC (Cisplatin plus Etoposide), and no recurrence was observed after 12 months.

Сравнительное исследование хромогранина А и хромогранина В у больных с нейроэндокринными опухолями желудка и поджелудочной железы

Introduction. Immunoenzyme assay of biochemical markers is one of the most important methods for examination of pa- tients with neuroendocrine tumors (NETs). Along with the generally accepted NET marker chromogranin A (CgA), another member of the granin family, chromogranin B (CgB), can serve as a complementary marker. Objectives. Analysis of CgB as an additional to CgA biochemical marker in the blood serum of patients with gastric and pancreatic neuroendocrine tumors. Materials and methods. We examined 79 patients with gastic ( n = 14) and pancretic ( n = 65) NETS, and 42 particularly healthy people, who were included in the control group. CgB and CgA were determined with ELISA method using the Human Chromogranin B (USCN, China) and Chromogranin A NEOLISA (Eurodiagnostica, Sweden) test systems. , Results. CgB levels in gastric and pancreatic NETs were significantly higher than in control group. CgB concentrations were independent of tumor spread and its biological activity. ROC analysis in common group of NETs relative to control group showed AUC for CgB = 0.869 and for CgA AUC = 0.82. According to results in common group of NET patients when used isolated, CgA and CgB have comparable diagnostic sensitivity, which increases in complex use to 82.5 %. In the group of NET patients with low levels of CgA (&lt;100 ng/ml), an increase in CgB concentration above the cut-off level (&gt;15.7 ng/ml) was observed in 53.6 % of cases. Conclusion. The combination of CgB and CgA in gastric and pancreatic NETs could increase the diagnostic efficacy of bio- chemical diagnostics. The received data confirms the significance of CgB as a complementary biomarker of NETs.

Perspectives on the diagnostic, predictive and prognostic markers of neuroendocrine neoplasms (Review).

Neuroendocrine neoplasms (NENs) are a heterogeneous group of rare tumors with different types of physiology and prognosis. Therefore, prognostic information, including morphological differentiation, grade, tumor stage and primary location, are invaluable and contribute to the formulation of treatment decisions. Biomarkers that are currently used, including chromogranin A (CgA), serotonin and neuron-specific enolase, are singular parameters that cannot be used to accurately predict variables associated with tumor growth, including proliferation, metabolic rate and metastatic potential. In addition, site-specific biomarkers, such as insulin and gastrin, cannot be applied to all types of NENs. The clinical application of broad-spectrum markers, as it is the case for CgA, remains controversial despite being widely used. Due to limitations of the currently available mono-analyte biomarkers, recent studies were conducted to explore novel parameters for NEN diagnosis, prognosis, therapy stratification and evaluation of treatment response. Identification of prognostic factors for predicting NEN outcome is a critical requirement for the planning of adequate clinical management. Advances in ‘liquid’ biopsies and genomic analysis techniques, including microRNA, circulating tumor DNA or circulating tumor cells and sophisticated biomathematical analysis techniques, such as NETest or molecular image-based biomarkers, are currently under investigation as potentially novel tools for the management of NENs in the future. Despite these recent findings yielding promising observations, further research is necessary. The present review therefore summarizes the existing knowledge and recent advancements in the exploration of biochemical markers for NENs, with focus on gastroenteropancreatic-neuroendocrine tumors.