Fetal antigen 1 (FA1) in the human pancreas: cell type expression, topological and quantitative variations during development

Monospecific rabbit anti-human fetal antigen 1 (FA1), was used to examine the distribution of FA1 during the development of the human fetal pancreas and liver using an indirect immunoperoxidase technique. FA1 was expressed by 94% of the glandular epithelial cells of the branching ducts in the pancreatic anlage at week 7 of gestation. This pattern changed during the development of the human pancreas, 64% of the glandular cells being FA1 positive at week 17 of gestation, decreasing to 11% in the infant (4 months after birth). In the infant and adults the FA1 expression was restricted to a subpopulation of -cells within the islets of Langerhans. Insulin immunoreactive cells were scattered throughout the epithelium of primitive branching pancreatic ducts at week 7 of gestation, well before the formation of islets. From the 7th through to the 17th week of gestation, FA1 was found in the cytoplasm of fetal hepatocytes, whereas no staining was observed in the liver from a 4-month-old infant. No FA1 expression was found in the epithelium of the developing gut. The present findings indicate that the glandular epithelial cells in the developing pancreas may serve as stem cells, which, if appropriately induced, may differentiate into endocrine cells. Fetal antigen 1 (FA1) may take part in or be a result of this differentiation.     

Intraperitoneal transplantation of pancreatic anlagen

To determine whether embryonic pancreatic anlagen transplanted to an intraperitoneal site in adult hosts grow, differentiate, and function, we implanted pancreas from embryonic day (E) 12.5 Lewis rat embryos into the omentum of adult Lewis rats or C57Bl/6J mice. E12.5 pancreatic anlagen were relatively undifferentiated except for the presence of condensing tubuloacinar cords. By 2 weeks after implantation, pancreatic anlagen transplanted into rats had enlarged and differentiated such that islets of Langerhans that stained positive for insulin could be delineated. Continued differentiation, as reflected by the presence of "ductal" islets connected to the duct epithelium, was observed at 6 weeks after implantation. At 15 weeks after implantation, "mature" islets had separated from the ducts. Electron microscopy showed eccentric dense bodies within clear vacuoles consistent with insulin granules. Little or no acinar tissue was present in developed anlagen. Within 5 weeks of pancreatic anlagen transplantation, levels of glucose in rats rendered diabetic by an injection of streptozotocin were normalized compared with levels in nontransplanted diabetic controls. Rat pancreatic anlagen underwent growth and development in the peritoneum of C57Bl/61 mice that received costimulatory blocking agents but not in the absence of costimulatory blockade. We concluded that whole E12.5 pancreatic anlagen undergo growth, differentiation, and function after intraperitoneal placement. Implantation of the embryonic pancreas, a "cellular" transplant, is followed by selective differentiation of islet compared with acinar components.     

Intrahepatic bile duct development in the rat: a cytokeratin-immunohistochemical study

The development of the intrahepatic bile ducts was studied in rats from day 12 of gestation until 10 days of age using three antibodies directed against cytokeratins in an immunohistochemical procedure on paraffin-embedded liver tissue. In adult rat liver, both hepatocytes and bile ducts were stained by the monoclonal anti-cytokeratin no. 8, whereas two polyclonal antibodies stained bile ducts only. Hepatocytes in developing rat liver were stained by monoclonal anti-cytokeratin no. 8 from day 12 of gestation on. On day 16, cells strongly immunoreactive for cytokeratin no. 8 were observed in a string of pearl-like arrangement around large vascular branches close to the liver hilum. Over the following days, similar structures appeared throughout the liver. Gradually, lumina were formed in these structures, again starting at the liver hilum and resulting in the formation of individual bile ducts. Immunoreactivity with the polyclonal antibodies was first detected in some of the string of pearl-like structures on day 19 and gradually increased until the intensity observed in adult rat liver was reached on day 1 after birth. Even on day 10, portal spaces still revealed more bile duct branches, rings of cells strongly positive for cytokeratin no. 8 and weakly positive with the polyclonal antibodies were present. It is concluded that the intrahepatic bile ducts develop from hepatocytes. The cells closest to large vascular spaces first become strongly positive for cytokeratin no. 8 and only later on acquire additional ("bile duct type") cytokeratins. This process starts at the liver hilum and spreads through the liver. Even at 10 days of age the bile duct system is still immature: around the smaller portal vein branches, rings of cells are still undergoing transformation into bile duct type cells. These data might be useful for reevaluation of pathologic phenomena.     

人胎胰腺GnRH免疫反应细胞

目的：探讨促性腺激素释的激素(GnRH)免疫反应细胞在人胎胰腺的存在部位和数量变化。方法：用免疫组织化学SABC法,对37例第10-32w人胎胰腺内的GnRH-IR细胞进行观察,并用体视方法分析其数量变化。结果：人胎胰腺GnRH-IR细胞出现于第13w,其数密度随胎龄增加而增大;分布于胰岛及外分泌部的腺泡上皮、导管上皮细胞间。位于胰岛的GnRH-IR细胞呈圆形、卵圆形或多边形。位于腺泡上皮细胞间的GnRH-IR细胞多为锥体形,外分泌部的GnRH-IR细胞均为开放型细胞。结论：胰腺GnRH-IR细胞于胚胎第13w出现,广泛存在于内、外分泌部,其数量随胎龄增加而增加。     

Marked diffuse dilations of the biliary tree associated with intrahepatic calculi,biliary sludges and a mucinous cyst of the pancreatic head in a 99-year-old woman

A 99-year-old woman was admitted to Shizuoka Shimizu Municipal Hospital because of fever and anasarca. Imaging and laboratory tests showed pneumonia, urinary tract infection, and cardiac failure. The patient died 20 days after admission. An autopsy revealed marked diffuse dilations of the biliary tree ranging from the lower common bile duct to intrahepatic bile ducts. Intrahepatic calcium bilirubinate stones and biliary sludges were recognized within the dilated bile ducts. A unilocular cyst (2 cm in diameter) was present in the pancreatic head adjacent to the lower common bile duct, and it appeared to compress the common bile duct. Histologically, the walls of the dilated biliary tree showed proliferation of peribiliary glands, fibrosis, and infiltration of lymphocytes and neutrophils (cholangitis). The lumens of the dilated biliary ducts contained neutral and acidic mucins, fibrinous materials, bacteria, neutrophils, and Aspergillus fungi, in addition to the calculi and sludges. The background liver showed atrophy (400 g). The pancreatic unilocular cyst was composed of mucous columnar cells with a few infoldings, and the pancreas also showed foci of mucinous duct hyperplasia and ectasia; the pathological diagnosis of the cyst was cystic dilations of a pancreatic duct branch (mucinous ductal ectasia or mucinous cyst). Other lesions included aspiration pneumonia, emaciation, atrophy of systemic organs, gastric leiomyoma, serous cystadenoma of the right ovary, and arteriosclerotic nephrosclerosis. The present case suggests that a mucinous cyst of the pancreas may compress the biliary tree and lead to marked diffuse dilations of the biliary tree. Alternatively, the dilations of the bile ducts may be associated with aging or may be of congenital origin. The dilated bile ducts may, in turn, give rise to bacterial and fungal cholangitis and formation of biliary sludges and intrahepatic calcium bilirubinate stones.     

Keratin immunohistochemistry in normal human liver. Cytokeratin pattern of hepatocytes,bile ducts and acinar gradient

Summary A panel of 2 polyclonal and 7 monoclonal antibodies directed against cytokeratins was tested on cryostat and paraffin sections of 14 normal human liver biopsies using an immunoperoxidase procedure. The staining characteristics of hepatocytes and bile ducts are reported. On cryostat sections, monoclonal antibodies directed against individual cytokeratins n^o8 and n^o18 stained both bile ducts and hepatocytes, whereas monoclonals anti-cytokeratin n^o7 and n^o19 exclusively stained bile ducts. The potential use of these 4 monoclonal antibodies in liver histopathology is briefly discussed. Monoclonal antibody anti-type II cytokeratins and the polyclonal rabbit anti-human keratin stained only bile ducts on both cryostat and paraffin sections. Using monoclonal antibody CAM 5.2 on paraffin sections, both bile ducts and parenchyma were positive. An acinar gradient was apparent in that zone 1 hepatocytes were more intensely stained. Moreover, a rim of hepatocytes around terminal hepatic venules and adjacent to subhepatic veins showed more intense staining. The same gradient could be seen in some paraffin sections stained with the monoclonals anti-cytokeratin n^o18 and KL1, and the rabbit polyclonal anti-keratin wide spectrum screening. The gradient is interpreted as reflecting quantitative differences in keratin content between hepatocytes. Polyclonal rabbit anti-human keratin is proposed as the most reliable antibody for identification of bile ducts in paraffin sections. The usefulness of reliable bile duct staining in several pathological conditions is emphasized.     

Simultaneous alpha-1-antitrypsin accumulation in liver and pancreas

Inclusions positive for periodic acid-Schiff, resistant to diastase, and immunoreactive to alpha-1-antitrypsin (AAT) were found in hepatocytes and pancreatic islet cells of a patient with clinical and pathologic features of AAT deficiency. Alpha-1-antitrypsin was detected in all pancreatic islets, and AAT-positive cells were observed in the excretory pancreatic ducts. These findings suggest that the pancreas synthesizes AAT and possibly serves as a "storage" place in AAT deficiency. Intercalated cells in the excretory pancreatic ducts may be an additional source of AAT.     

Intraductal papillary–mucinous neoplasm of the pancreas associated with polycystic liver and kidney disease

A case of intraductal papillary–mucinous neoplasm (IPMN) of the pancreas associated with polycystic liver and kidney disease is reported. A 63-year-old man was admitted to hospital with heartburn and upper abdominal pain. CT indicated multiple cysts in the liver and bilateral kidneys, and also showed multiple cystic lesions in the pancreatic head. Pancreatoduodenectomy was performed, and a pathological diagnosis of intraductal papillary–mucinous adenoma of branch duct type of the pancreas was made. He died of renal failure and hepatic failure due to recurrent cholangitis 12 years after the operation. An autopsy confirmed multiple cysts in the liver and kidneys. Multiple hepatic cysts were lined by biliary-type epithelium with no connection to the bile ducts. Peribiliary cysts were observed, and histology of the liver indicated von Meyenburg complexes. The kidneys showed that the cysts originated from all segments of the renal tubule. The autopsy findings suggested that the patient had suffered from autosomal-dominant polycystic kidney disease (ADPKD), but he did not have a family history of ADPKD. This case demonstrates that IPMN of the pancreas can occur as an extrarenal complication in patients with polycystic liver and kidney disease.     

Nonsyndromatic paucity of intrahepatic bile ducts in congenital syphilis. A case report

The first case of nonsyndromatic paucity of the intrahepatic bile ducts is reported in congenital syphilis. The patient, a 2-week-old female, was born at the 31st week of gestation, weighing 1,910 g. She had a high titer of IgM antibody to Treponema pallidum and sera from both parents also showed a positive reaction in the hemagglutination test for Treponema pallidum. The patient had hepatosplenomegaly and increasing jaundice, and died of respiratory failure on the 70th hospital day. Autopsy examination revealed paucity of the intrahepatic bile ducts, prominent giant cell transformation of hepatocytes, cholestasis and extramedullary hematopoiesis of the liver. The ratio of the number of intrahepatic bile ducts to that of the portal tracts was approximately 0.2:1. There was marked proliferation of atypical bile ductules at the margin of the portal tracts. The exact relation of this paucity to Treponema pallidum remains unknown.     

Serotonin immunoreactivity in pancreatic endocrine neoplasms (carcinoid tumors).

Primary serotonin secreting pancreatic endocrine neoplasms (carcinoid tumors) are extremely rare and may be associated with manifestations of the carcinoid syndrome. Two cases of primary carcinoid tumor of the pancreas with liver metastases showed clinical and biochemical features of the carcinoid syndrome. Both cases demonstrated strong positive immunoreactivity for serotonin within the tumor cells. In an attempt to determine the relationship between pancreatic carcinoid tumors and other pancreatic endocrine neoplasms, immunostains for serotonin were performed on 11 additional islet cell tumors and on non-neoplastic pancreatic tissues. These cases showed serotonin immunoreactivity within islet cell tumors (36%). In addition, focal staining for serotonin was present in non-neoplastic ducts and ductules (88%), acini (22%), and islets of Langerhans (33%). Based on these observations, specific criteria are suggested for the diagnosis of primary pancreatic carcinoid tumor.     

Dynamic distribution of ERK,p38 and JNK during the development of pancreatic ductal adenocarcinoma

We recently discovered that oncogenic c-kit is highly expressed concomitantly with the development of pancreatic ductal adenocarcinoma (PDAC). Since oncogenic c-kit may activate major pathways of protein tyrosine phosphorylation, we decided to investigate this issue in the major protein phosphorylation cascades. In normal pancreas labeling with antiphosphorylated ERK1/2 (pERK1/2) antibody was mainly confined to islets of Langerhans in close overlapping with insulin containing cells. Phosphorylated p38 (pp38) showed a similar pattern of distribution, while only weak labeling was evident for pJNK and no labeling of pMEK was observed. As expected, general ERK1/2 (gERK1/2), general p38 (gp38), general JNK (gJNK) as well as general MEK (gMEK) were all evident in islets of Langerhans and in the exocrine tissue. In early development of PDAC, pERK1/2 and pp38 retained their localization in islets of Langerhans. Intensive staining of pERK1/2 was also evident in the cancerous ducts, while the labeling with antibodies to pp38 was more moderate. While pJNK staining in islets of Langerhans was weak, with no labeling in the cancerous ducts, antibodies to gJNK revealed intensive staining suggesting the weak staining of pJNK is not due to the lack of the enzyme. In a more advanced stage of PDAC the carcinomas were clearly stained with pERK1/2 and pp38, while moderate staining with pJNK was also evident. In liver metastases, the cancer cells were heavily labeled with all three phospho-MAPKs. It should be noted that the localization of all three kinases was mainly in the cell nuclei. In the more advanced stage of PDAC, heavy labeling was evident using antibodies to gERK1/2, gp38, gJNK and gMEK. However, no labeling to pMEK was evident in parallel sections. Our data suggest that both in normal and cancerous pancreas, most of the MAPK activities are located in islets of Langerhans and cancerous ducts. It is suggested that using inhibitors to protein kinases may attenuate the progression of the disease.     

Development of colonic and pancreatic endocrine tumours in mice expressing a glucagon-SV40 T antigen transgene

We report the histological, immunohistochemical and ultrastructural changes in mice containing a chimeric glucagon-simian virus 40 T antigen (SV40Tag) gene. Transgene expression was detected in endocrine cells of pancreas, small and large intestine. Hyperplasia of glucagon-containing cells developed in pancreas and large bowel by gestational day 19. In large bowel, hyperplastic cells increased in number postnatally and invasive carcinomas were identified at 4 weeks; several animals had lymph node metastases. In contrast, no pathology was detected in the small bowel in any of the transgenic mice. Colonic tumours expressed SV40Tag, proglucagon-derived peptides and peptide YY (PYY); scattered cells contained cholecystokinin or glycoprotein hormone -subunit. Somatostatin or serotonin was also detected in some tumours. By electron microscopy, the colonic tumours retained features of endocrine differentiation, but secretory granules were smaller than those of non-tumorous intestinal glucagon-producing L cells. In postnatal pancreas, atypical cells containing SV40Tag and glucagon were initially clustered at the periphery of islets; this atypical hyperplasia progressed to neoplasia by 11–12 weeks. Some neoplastic pancreatic cells contained glucagon, PYY or vasoactive intestinal peptide immunopositivity, but most were negative for all peptides; they contained immunoreactivity for tyrosine hydroxylase and by electron microscopy, pancreatic tumour cells had neuronal features. Pancreatic polypeptide was not detected in the non-tumorous islets of transgenic animals. This line of transgenic mice provides a model for the analysis of endocrine tumour progression in the gut and pancreas.     

Diabetes mellitus and fatty liver in a cow: case report

A 5-year-old Holstein–Friesian cow was referred to the Veterinary Clinic of Shiraz University in May 2003 with a history of continuous weight and milk loss for 32 days after calving. On clinical examination the animal was moderately depressed. Pulse, respiratory rate and temperature were normal. Rumen motility was decreased in strength and rate. The faeces were dry, firm and scanty. Polyuria and polydypsia were noticed. The results of biochemical analysis revealed a significant rise in urine ketones and glucose. The blood glucose, cholesterol, triglyceride and -hydroxybutyrate levels were considerably above the normal range. At necropsy the liver was enlarged, pale, and yellow and in the cut section the liver parenchyma had a fatty quality. The gross anatomy of the pancreas and kidneys was normal. On microscopic examination severe fatty change in the liver was observed. Fatty change in the acinar tissue of the pancreas was also seen. The number and size of islets were reduced. On the basis of characteristic staining reaction, depletion of beta cells was diagnosed. According to the history, clinical signs and laboratory findings, a combination of diabetes mellitus and fatty liver was diagnosed.     

Ghrelin in the fetal pancreas - a digital quantitation study

Ghrelin is a hormone produced by specialized neuroendocrine cells located in the fetal pancreas. In the adult, ghrelin has multiple effects, but in the fetus the role of ghrelin and the distribution of ghrelin-producing cells is not well documented. The aim of this study was to describe and quantitate the number of ghrelin positive cells in the pancreas during gestation. The material consisted of pancreatic tissue from 19 fetuses at different gestational ages. Immunohistochemical staining was performed, and the expression was quantitated using an automated digital image analysis system. The results showed ghrelin-producing cells as scattered single cells in ductular structures and acini throughout the gestation. From midgestation they were also found in the periphery of the islets as a rim of cells. A tendency towards a high ghrelin expression during early gestation and a stable expression from midgestation to term was observed. In conclusion, the effects of fetal ghrelin are not fully understood, but the varying distribution of ghrelin positive cells indicates different effects of ghrelin during development.     

Renal-hepatic-pancreatic dysplasia: an autosomal recessive malformation.

We report two brothers with a cystic malformation of the kidneys, liver, and pancreas. In both cases the malformation was fatal and the children died shortly after birth. The pathological findings, consisting of multicystic dysplastic kidneys, dilated and dysgenetic bile ducts, dilated pancreatic ducts, and polysplenia, correspond to those reported by Ivemark as renal-hepatic-pancreatic dysplasia. Many polymalformation syndromes include cystic affectation of these three organs, so this syndrome could be an isolated entity or a final common pathway of response of these organs to a variety of developmental disturbances, which could also include splenic abnormalities. We propose an autosomal recessive pattern of inheritance for renal-hepatic-pancreatic dysplasia.     

NONSYNDROMATIC PAUCITY OF INTRAHEPATIC BILE DUCTS IN CONGENITAL SYPHILIS. A Case Report

The first case of nonsyndromatic paucity of the intrahepatic bile ducts is reported in congenital syphilis. The patient, a 2-week-old female, was born at the 31st week of gestation, weighing 1,910 g. She had a high titer of IgM antibody to Treponema pallidum and sera from both parents also showed a positive reaction in the hemagglutination test for Treponema pallidum. The patient had hepatosplenomegaly and increasing jaundice, and died of respiratory failure on the 70th hospital day. Autopsy examination revealed paucity of the intrahepatic bile ducts, prominent giant cell transformation of hepatocytes, cholestasis and extramedullary hematopoiesis of the liver. The ratio of the number of intrahepatic bile ducts to that of the portal tracts was approximately 0.2:1. There was marked proliferation of atypical bile ductules at the margin of the portal tracts. The exact relation of this paucity to Treponema pallidum remains unknown. ACTA PATHOL JPN 38: 1061 ∼ 1068, 1988.     

大鼠胰组织块的三维重建及形态学测量

目的:观察大鼠胰组织块内部各结构的空间位置关系,并描述胰岛的立体形态学特征.方法:经过长序列连续切片、H-E染色、切片图像数字化及配准等步骤,建立正常大鼠胰组织块的数字化图像数据集,对其内部的动脉、静脉、导管及胰岛进行分割及三维重建,并对重建结果进行测量及分析.结果:所得数字化图像数据集分辨率高,配准准确,再现了原组织的结构,重建结果可在三维空间内自由观察.根据重建结果测量多种形态学指标.胰腺泡及结缔组织占胰总体积的绝大部分,胰岛占1.125%.结论:胰内动脉、静脉分支自主干垂直发出后,互相伴行,先于胰表面,随后向小叶深部发出分支.胰导管走行独立,分支间存在吻合.某些部位动脉、静脉与导管并行,形成三联管结构.胰岛与导管系统关系密切.     

Licht- und Elektronenmikroskopische Leberbefunde beim Cerebro-Hepato-Renalen Syndrom nach Zellweger (Peroxisomen-Defizienz)

Summary For histologic differential diagnosis of fibrotic and cirrhotic liver changes in early infancy Zellweger's cerebro-hepato-renal syndrome has to be considered. A case is reported where a female newborn failed to thrive and developed severe muscular hypotonia, defective intelligence, and seizures. At an age of 12 months an open biopsy was taken from the enlarged liver. Biochemical tests for disorders of carbohydrate metabolism were negative. The light microscopic examination showed severe fibrosis, lobular disarray, and single cell necroses. Neither siderin nor bile pigment could be detected. There were no changes of the small bile ducts and no signs of inflammation — the child died at 18 months. A postmortem needle biopsy of the liver showed the advanced stage of micronodular cirrhosis.The electron microscopic examination of the first biopsy revealed hypertrophy of smooth endoplasmic reticulum, probably induced by anticonvulsive drugs. The mitochondria showed an increase in the density of their matrix and in the number of sometimes tubular cristae. In accordance with reports in literature, no regular peroxisomes (microbodies) could be found in the hepatocytes. Only a very few profiles with an average diameter of 0.16 m looked like rather small peroxisomes. The cytoplasmic volume fraction of those organelles, however, amounted to 3×10^–5 which is only 1/500 of the normal value in human hepatocytes.Although the significance of the absence of peroxisomes is not yet definitively proved, the term Peroxisome Deficiency is proposed as a name which probably is more closely related to the pathogenetic mechanism than the hitherto usual designation cerebro-hepato-renal syndrome.

     

Neonatal hemochromatosis--report of an autopsy case.

A case of neonatal hemochromatosis in a 3-hour-old male is described. He presented with hypotonia, mild jaundice, and respiratory difficulty immediately after birth. He had no evidence of congenital infection, immune-related hemolysis or exogenous iron uptake. Postmortem examination revealed abnormal facial features. The organs were of normal weight for his age except a small liver and lungs, and a large spleen. The most prominent changes were in the liver and pancreas. The liver was coarsely nodular and fibrotic. The lobular architecture was totally distorted by innumerable multinucleated giant cells, loss or collapse of the hepatocytes, and diffuse fibrosis. A large amount of hemosiderin was seen in the liver, pancreatic acini and thyroid follicular cells. Scanty amount of hemosiderin was also found in the myocardial fibers and renal tubular cells. The pancreas showed hyperplasia and hypertrophy of the islets. The spleen showed severe congestion and a moderate extramedullary hemopoiesis but no deposits of hemosiderin. This patient had three siblings died in neonatal period, one of which had clinical features of neonatal hemochromatosis.