Search

BACKGROUND: No consensus exists concerning dosage and duration of prophylactic hepatitis B immunoglobulin and lamivudine for prevention of hepatitis B recurrence after liver transplantation (LT). METHODS: Lamivudine was discontinued 12 months after LT, maintaining hepatitis B immunoglobulin prophylaxis in eight patients who received lamivudine treatment before LT. RESULTS: At LT, six patients were serum hepatitis B virus (HBV)-DNA negative, whereas two patients had low serum HBV-DNA levels. Hepatitis B surface (HBs) antigen and hepatitis B core antigen stained positively by immunohistochemistry in all hepatectomy specimens. All patients remained recurrence free during the 12 months on combination therapy with normal liver histological examination and negative HBs and HB core staining on biopsy specimens. No relapse occurred after lamivudine withdrawal during a median follow-up of 17.5 months (normal transaminases, negative serum HBs antigen, and HBV-DNA). CONCLUSIONS: Discontinuation of lamivudine 12 months after LT is feasible and safe even in patients with ongoing low viral replication at LT, providing adequate prophylaxis with hepatitis B immunoglobulins.

BACKGROUND/PURPOSE: Permanent hyperinsulinemic hypoglycaemia in infancy (PHHI)I is a severe disease that leads to brain damage. Since 1989, pathologists have identified 2 different forms of the disease: a diffuse form (DiPHHI) and a focal form (FoPHHI). The purpose of this study was to adapt surgical techniques in case of FoPHHI to cure these infants without risk of diabetes. METHODS: All patients with PHHI underwent pancreatic venous sampling (PVS) and elective partial pancreatectomy (EPP). Molecular biology and immunohistochemistry were used to ascertain that FoPHHI was a different disease from DiPHHI. RESULTS: 45 EPPs were performed, guided by PVS and peroperative pathology. The lesions were 17 in the head, 4 in the isthmus, 6 in the body, 15 in the tail of the pancreas. Age at surgery ranged from 25 days to 4 years. Two patients already had been operated on elsewhere, and the focal lesion could be found at second operation. All 45 patients except one, were cured with euglycemia at both fasting and hyperglycaemic tests. Molecular biology has shown a specific anomaly in FoPHHI, which never has been encountered in DiPHHI. CONCLUSIONS: PHHI is not a homogeneous disease. In one third of cases, only a small amount of endocrine pancreas is abnormal, and conservative surgery is mandatory. The pre- and perioperative conditions to point out the focal pancreatic lesion are described.

Hyperinsulinism is a heterogeneous disorder characterised by severe hypoglycaemia due to an inappropriate oversecretion of insulin. In a personal series of 175 patients investigated for hyperinsulinaemic hypoglycaemia over the last 20 years, we review clinical presentations, molecular studies and therapeutic management of hyperinsulinism. There were 98 neonatal-onset patients, including 86 permanent hyperinsulinism and 12 transient forms, 68 with infancy-onset and nine with childhood-onset. Hyperammonaemia was found in 12 out of 69 patients tested, 4 neonates and 8 infants. Neonates were clinically more severely affected than infants. Diagnosis of infancy-onset hyperinsulinism was often delayed because of less profound hypoglycaemia and better tolerance to hypoglycaemia. Neonates required higher rates of i.v. glucose than infants to maintain normal plasma glucose levels (16 mg/kg per min versus 12 mg/kg per min). Only 16% of neonates were diazoxide-sensitive compared to 66% of the infants. Neonates with hyperammonaemia or transient hyperinsulinism were diazoxide-sensitive. Most neonates were pancreatectomised whereas 65% of the infants were treated medically. Among surgically-treated patients, 47% had a focal adenomatous hyperplasia (31 neonates and 13 infants) and 53% a diffuse form of hyperinsulinism (39 neonates and 11 infants). Diazoxide-responsiveness in the focal and diffuse forms did not differ in both neonates and infants; it depended only upon the age of onset of hypoglycaemia. One or two mutations, SUR1 or KIR6.2, were found in 41 of 73 neonates who were investigated and in 13/38 infants using polymerase chain reaction-single strand conformational polymorphism analysis of both genes. Almost all patients with SUR1 (38/41) or KIR6.2 (5/7) mutations were resistant to diazoxide. Ten patients with hyperinsulinism-hyperammonaemia syndrome had a mutation in the glutamate dehydrogenase gene (three neonates and seven infants) after reverse transcriptase-polymerase chain reaction and sequence analysis of cDNA. No mutation was found by polymerase chain reaction-single strand conformational polymorphism in the glucokinase gene. Eight of nine patients with childhood onset hyperinsulinism were treated surgically and histological examination confirmed an adenoma in each case. Conclusion: the clinical severity of hyperinsulinism varies mainly with age at onset of hypoglycaemia. The heterogeneity of hyperinsulinism has major consequences in terms of therapeutic outcome and genetic counselling.

Beyond the 2 classical forms of congenital hyperinsulinism, focal and diffuse, we report our experience on the surgical treatment of atypical forms. We define 2 subtypes among these atypical forms of hyperinsulinism: in case of a giant focal form the surgical strategy is the same as in focal forms. In case of hyperinsulinism caused by a mosaic, our experience suggests the benefit of a limited resection from the tail to the body of the pancreas.

BACKGROUND/AIMS: Patients with end-stage liver disease due to chronic hepatitis B infection in whom a YMDD escape hepatitis B virus (HBV) mutant has emerged under lamivudine treatment are generally denied liver transplantation (OLT). METHODS: We report the case of a male patient who was started on prophylactic treatment with lamivudine in the context of recurrent episodes of HBV reactivation during high dose immunosuppressive therapy for relapsing severe pulmonary sarcoidosis. RESULTS: Following the emergence of a YMDD escape mutant virus under lamivudine treatment, he developed subacute liver failure requiring liver transplantation. The patient was treated with a combination of intravenous hepatitis B immune globulin (HBIG) which was started perioperatively and also continued lamivudine after OLT. Twelve months after OLT, there was no evidence of HBV reinfection of the liver graft with the use of HBIG and lamivudine. CONCLUSIONS: This observation suggests that emergence of the YMDD mutation is not a contra-indication to OLT, providing adequate immunoprophylaxis using HBIG and lamivudine combination therapy.

We report the case of a child who developed, 2 yr after orthotopic liver transplantation (OLTx) for biliary atresia, a multi-focal hepatic tumor with lymphonodular metastases, identified as an Epstein-Barr virus (EBV)-associated leiomyosarcoma. Chemotherapy was given without tumor response. Subsequently, slow growth of the tumor was observed. Immunosuppression was tapered and stopped 9 yr after transplantation. At the present time, 12 yr after the discovery of the first hepatic lesions, the patient is alive and completely symptom-free, the abdominal masses are stable, and liver function tests are completely normal. Smooth muscle tumors are increasingly recognized in children with various immunodeficiencies occurring after organ transplantation. This unusual evolution of a clinically aggressive tumor into a stable disease after restoration of immunity confirms that the immune status of the patient is a crucial factor.

Persistent hyperinsulinaemic hypoglycaemia of infancy (PHHI) is a heterogeneous disorder characterized by profound hypoglycaemia due to inappropriate hypersecretion of insulin. An important diagnostic goal is to distinguish patients with a focal hyperplasia of islet cells of the pancreas (FoPHHI) from those with a diffuse abnormality of islets (DiPHHI), because the management differs significantly. The intriguing similarity between islet cell hyperplasia and tumourigenesis prompted us to investigate whether the imprinted genes in the 11p15 region are involved. Results showed that diffuse forms are caused by constitutional homozygous or compound heterozygous mutations of the SUR1 gene. In contrast, focal forms are caused by loss of the maternally inherited 11p15 region, resulting in both loss of the maternally expressed tumour suppressor genes accounting for hyperplasia and somatic reduction to hemizygosity or homozygosity of the paternally inherited SUR1, limited to the lesion. Thus, this somatic disorder, which leads both to β-cell proliferation and to hyperinsulinism, can be considered the somatic equivalent, restricted to a microscopic focal lesion, of constitutional uniparental disomy associated with unmasking of a heterozygous parental mutation.

The syndrome of persistent hyperinsulinaemic hypoglycaemia of infancy (PHHI) was described more than 40 years ago by Mc Quarrie.1 Despite the inordinate amount of interest in this syndrome, the pathogenesis of the disease has not yet been completely elucidated. For decades, the disease has been ascribed to nesidioblastosis. This term, first coined by Laidlaw, describes the persistence of a diffuse and disseminated proliferation of islet cells budding off from ducts.2 The concept of nesidioblastosis as the underlying condition of hyperinsulinism is still deeply rooted in the mind of many clinicians, although it has been questioned by several authors.3-5 Indeed, observations based on quantitative immunohistochemical investigations have shown that nesidioblastosis is a common feature of the pancreas in normoglycaemic neonates and infants (fig 1).3 Figure 1 Nesidioblastosis in the pancreas of a normoglycaemic infant. B cells budding off from ducts stained by an anti-insulin antibody (immunoperoxidase; original magnification, ×140). Progress in genetics and molecular biology has increased our understanding of the syndrome. By means of linkage analysis, hyperinsulinism in familial cases has been mapped to chromosome 11.6 Genetic molecular analyses have demonstrated mutations of the genes encoding both the sulphonylurea receptor7-10 and the Kir 6.2 subunit,11 as well as maternal loss of the imprinted gene at the 11p15 region, which might explain the insulin hypersecretion characteristic of this syndrome.12-14 In addition, physiological analysis has demonstrated the absence of functional ATP dependent potassium channels in some of these infants with hypoglycaemia.10 15 Several studies have clearly demonstrated the existence of two forms of PHHI. One corresponds to a focal pancreatic adenomatous hyperplasia (focal PHHI) and the other is characterised by a diffuse β cell abnormality (diffuse PHHI).16-18 Up until now, these two forms could not be differentiated by clinical or biochemical data, although …

BACKGROUND & AIMS: Drug-induced immunoallergic hepatitis typically affects a minority of patients exposed to a particular drug. Its rarity is believed to be due to metabolic or immunologic idiosyncrasy. The presence of an immunologic idiosyncrasy might imply an HLA association. Previous studies reporting an HLA association of drug-induced hepatitis included only small numbers of patients and used serological HLA typing. METHODS: We studied 35 patients with biopsy-documented amoxicillin-clavulanate-induced hepatitis. HLA-A and -B were typed using alloantisera and compared with those of 300 controls (volunteer bone marrow donors). HLA-DRB and -DWB were typed by polymerase chain reaction-line probe assay, with 60 volunteer bone marrow donors serving as controls. RESULTS: The study group was characterized by a higher frequency of DRB1*1501-DRB5*0101-DQB1*0602 haplotype (57.1% vs. 11.7% in controls, P < 0.000005; after correction for the large number of comparisons, P < 0.0002). Patients with DRB1*1501-DRB5*0101-DQB1*0602 haplotype were more likely than patients without it to have a cholestatic (70% vs. 60%) or mixed (30% vs. 13%) than a hepatocellular pattern of hepatitis (0% vs. 27%) (P < 0.05). CONCLUSIONS: Amoxicillin-clavulanate-induced hepatitis is associated with the DRB1*1501-DRB5*0101-DQB1*0602 haplotype. The data support the view that an immunologic idiosyncrasy, mediated through HLA class II antigens, plays a role in the pathogenesis of drug-induced immunoallergic hepatitis. HLA association has a limited impact on the expression of hepatitis.

Long-term diabetes mellitus (DM) is characterized by widespread alterations of basal lamina (BL). The purpose of the present work was to verify whether the lung is also a target organ damaged in DM. Electron microscopy was performed on lung and kidney samples (autopsic material) from 6 diabetics and 6 control subjects studying the thickening of BL of different structures (alveolar epithelial BL, endothelial capillary BL, both fused BL, BL of the glomerular capillary endothelium and BL of the renal tubules). The results were as follows: (1) alveolar epithelial BL (mean ± SD) = 121 ± 11 nm in controls and 176 ± 27 nm in diabetics (p < 0.01), (2) endothelial capillary BL = 164 ± 14 nm in controls and 223 ± 27 nm in diabetics (p < 0.001), (3) both BL fused = 222 ± 23 nm in controls and 316 ± 62 nm in diabetics (p < 0.01), (4) BL of the glomerular capillary endothelium = 374 ± 44 nm in controls and 626 ± 249 in diabetics (p < 0.05) and (5) BL of the renal tubules = 602 ± 94 nm in controls and 1,083 ± 376 nm in diabetics (p < 0.05). All parts of the lung are equally affected by DM. The thickening of BL is of the same magnitude in lung and kidney. There is no relationship between the thickening of the lung BL and the known duration and type of DM.

The case of a young woman with GB virus C/hepatitis G virus (GBV-C/HGV) infection and with a severe exacerbation of chronic hepatitis of unknown etiology during pregnancy is described. In the offspring, severe neonatal hepatitis with subsequent mild chronic liver disease of at least 16-month duration was followed by the development of antibodies to the envelope protein (E2) of GBV-C/HGV, suggesting that the child was recovering from GBV-C/HGV infection. There was an improvement in clinical and biochemical parameters in the mother following delivery and alpha-interferon therapy was associated with a transient biochemical response.

Recurrence of autoimmune hepatitis after orthotopic liver transplantation and after reduction in immunosuppressive treatment is reported in a 43-year-old man. Diagnosis was based on biological, serological and morphological data, as well as on a complete biological recovery following steroid resumption. Recurrence was observed only 4 months after transplantation, when an abrupt increase in liver function tests was observed in association with a mainly lobular hepatitis in liver biopsy. Such a histological appearance has been previously described in the case of recurrence after immunosuppressive treatment weaning in non-transplanted patients. In contrast, this histological feature has not been reported in the rare transplanted patients in whom such recurrence has been observed. Thereafter, the evolution of the disease in our patient remained slow, likely dampened by the administration of cyclosporine and azathioprine. Morphological features are detailed, and importance of immunosuppressive therapy in patients transplanted for end-stage autoimmune hepatitis is emphasized.

The mechanism(s) by which rat immature enterocytes exhibit increased responsiveness to insulin before weaning is unknown. Therefore, we have analyzed the distribution, ontogeny, and molecular properties of insulin receptors (IR) and of related substrates in immature and mature enterocytes. IR were studied by radioligand binding assays, cross-linking labeling, immunohistochemistry, and in vitro phosphorylated substrates by immunoprecipitation. Regardless of age, 125I-insulin binding to IR was five times higher in crypt cells than in villus cells and two times higher in the ileum than in the jejunum. Binding capacity to villus cells from sucklings (day 14) exceeded three times that of older animals (day 30 and day 60). Scatchard analysis of equilibrium binding data confirmed an age-related decrease in low- and high-affinity receptor classes without change in affinity constants. In concordance, both alpha- and beta-IR subunits were more abundant in immature than in mature membranes. In vitro, insulin elicited the phosphorylation of three membrane proteins (96, 60 and 42 kDa), whose signals were virtually inhibited by preincubating membranes with antireceptor monoclonal antibodies. By immunoprecipitation, the 60-kDa signal was rapidly detected as a tyrosine-phosphorylated protein, expressed in mature and immature membranes, and identified as a receptor substrate phosphorylated in vitro by the IR tyrosine kinase. In conclusion, 1) increased responsiveness of rat immature enterocytes to insulin could be related to high membrane concentrations of IR and 2) normal rat enterocytes express a 60-kDa phosphotyrosine protein identified as a direct substrate of the IR tyrosine kinase.

Introduction: Papillary thyroid cancer (PTC) is the commonest endocrine malignancy. PTC frequently carries several alterations in genes coding for proteins that activate the mitogen-activated protein kinase (MAPK) signaling pathway, which plays a key role in the regulation of cell growth, differentiation, and survival. The present study aims to investigate the MAPK signal transduction pathway [extracellular-regulated kinase (ERK), Jun N-terminal Kinase (JNK) and p38] involved in tumorigenesis of PTC. Experimental procedures: Twenty samples of PTC and its follicular variant (11 classic PTC, 9 follicular variant of PTC) were retrieved from the UCL biolibrary. Corresponding paraffin blocks were selected for immunohistochemical analysis of total and phosphorylated forms of ERK, JNK and p38. Percentage of immunolabeled cells in tumors was graded as: slight staining in less than 10% of tumor cells; mild staining in 10-30%; moderate staining in 30-50% and abundant staining in more than 50 % of tumor cells. In addition, frozen sections of the same tumors were solubilized to perform Western blots of total and phosphorylated p38. Results: ERK activation was seen as nuclear immunolabeling in 15/ 20 PTC cases in less than 10% of tumor cells. JNK activation was seen in 7/20 cases in less than 10% of tumor cells as nuclear and membranous immunolabeling. However, total ERK and total JNK was seen as nuclear and cytoplasmic immunolabeling in 10/20 and 16/20 cases respectively in more than 50% of tumor cells. p38 MAPK phosphorylation was seen as abundant nuclear and cytoplasmic immunolabeling in 11/20 cases (6 follicular variants of PTC and 5 classic PTC). Total p38 was immunolabeled in 14/20 cases. A one-way ANOVA test showed significant difference between the ERK, JNK and p38 phosphorylation (p By Western blotting, p38 was detected in all twenty PTC samples and its phosphorylated form was detected in 17/20 samples. The signals were much stronger in the follicular variants of PTC as compared to classic PTC. p38 was also detected in normal thyroid tissue but not its phosphorylated form. Conclusions: There is variable expression and activation of the MAPK pathways in PTC but p38 appears to be activated in a larger proportion of PTC than ERK or JNK. The role of p38 in tumorigenesis of PTC and its variable expression in different variants of PTC will be further investigated. The molecular profiling of PTC should help to better understand the altered biological pathways involved in the genesis of this cancer. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C148. Citation Format: Monika Lamba Saini, Birgit Weynand, Jacques Rahier, Michel Mourad, Marc Hamoir, Etienne Marbaix. Expression and activation of p38 in papillary thyroid carcinoma. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C148.

Lefebvre S, De Paepe L, Abs R, Rahier J, Selvais P, Maiter D. Subcutaneous octreotide treatment of a growth hormone-releasing hormone-secreting bronchial carcinoid: superiority of continuous versus intermittent administration to control hormonal secretion. Eur J Endocrinol 1995;133:320–4. ISSN 0804–4643 Diagnosis of ectopic acromegaly was made in a 21-year-old female patient who 3 years before had undergone a right pneumectomy for a disseminated bronchial carcinoid. Plasma growth hormonereleasing hormone (GHRH) concentrations were markedly elevated (6440 ng/l; normal value S Lefebvre, Division of Rheumatology, Clinique du Refuge, Rue du Couvent 39, B-7700 Mouscron, Belgium

In situ hybridization represents a major advance in the study of gene expression and, thus, in the evaluation of cellular function in histological sections. The availability of oligonucleotide probes labelled with biotin and sensitive immunohistochemical detection systems makes the study of different types of mRNA by in situ hybridization easier. However, a large number of protocols have been reported, which is sometimes confusing. The present study analyses quantitatively the influence of each important step of in situ hybridization on the staining intensity of rat proinsulin mRNA. The aim was to optimize technical conditions, to make the method sensitive and to evaluate its reproducibility for proinsulin mRNA detection and measurements. The duration of fixation and the digestion have an important impact on the results. The optimal digestion time depends on the fixation. With a digestion of 30 micrograms ml-1 proteinase K for 12 min at 37 degrees C, the optimal fixation time was 24 h. Section thickness also influences the staining intensity. The intensity of the staining increases as the section thickness increases from 3 to 5 microns before slowly decreasing. A weak paraformaldehyde post-fixation (0.4% for 20 min) gives best results in comparison to a stronger post-fixation (4% for 10 min). An increase of probe concentration leads to a higher specific labelling, reaching a plateau at 800 ng ml-1. Hybridization temperature (37-42 degrees C) exerts little influence. However, the temperature of the washes and the immunodetection system have a major effect on the intensity of labelling.(ABSTRACT TRUNCATED AT 250 WORDS)

Paediatric orthotopic liver transplant recipients may develop chronic hepatitis after surgery. To investigate the role of hepatitis C virus in this pathology a cohort of 249 paediatric orthotopic liver transplant recipients was studied. Sixteen children (6.4%) were found to have chronic hepatitis C virus hepatitis after orthotopic liver transplantation. All but one of them had serum transaminase values which were persistently raised two to eight times the upper limit of normal. Thirteen were positive for both serology and serum hepatitis C virus RNA. Serum hepatitis C virus RNA detection occurred five to 33 months before hepatitis C virus antibodies. Liver tissue hepatitis C virus RNA and hepatitis C virus core antigen were detected in five. In one patient, tissue hepatitis C virus core antigen was detected when other tests for hepatitis C were negative. Two patients had positive human cytomegalovirus serum antibodies and RNA before transplantation. Although serum hepatitis C virus RNA was not detected after transplantation, serum enzyme immunosorbent assay and tissue core antigen were still detectable in both patients. In another child, serum hepatitis C virus RNA was positive and hepatitis C virus core antigen was found on a liver biopsy specimen but antihepatitis C virus antibodies were negative as well as liver hepatitis C virus RNA. No patient developed severe liver disease or cirrhosis during a follow up of up to 72 months. It is concluded that hepatitis C virus is a significant cause of morbidity after paediatric orthotopic liver transplantation. Diagnosis cannot rely on serological testing only. The patients remained stable on follow up, but longer prospective histological studies remain necessary to establish prognosis.

The evolution from a characteristic picture of autoimmune chronic hepatitis type I to primary biliary cirrhosis is reported in a middle-aged woman. The initial diagnosis of autoimmune chronic liver disease was based on clinical, bio-serological and histological grounds. It was further confirmed by complete remission following immunosuppressive treatment and prompt relapses at the time of therapy withdrawal. After 7 years, the characteristics of liver disease were altered with increased biochemical cholestasis and serum IgM concentrations, positivation of previously negative anti-M2 antimitochondrial antibodies and the appearance of a typical histological picture of stage I primary biliary cirrhosis. We believe that this is the first reported case in which autoimmune chronic hepatitis highly responsive to immunosuppressive therapy has been followed by the development of a characteristic picture of primary biliary cirrhosis.

A prospective trial was conducted to assess the efficacy of induction immunosuppression with antilymphocyte monoclonal antibodies in 129 primary liver transplant patients who were randomly divided into three groups according to immunosuppression during the first 10 days post-OLT: triple drug therapy only (TDIS: cyclosporine, steroids, azathioprine) (group I: n = 42); TDIS with a 10-day course of OKT3 (group II: n = 44); and LO-Tact-1 (anti-IL-2 receptor mAb) (group III: n = 43). Biopsy-proved acute rejection (AR) was treated using the same biopsy-guided protocol in the 3 groups. One-year patient survival rates were 67%, 84%, and 93% in groups I, II, and III, respectively (I vs. II, NS; I vs. III, P = 0.001; II vs. III, P = 0.044). Incidences of AR were studied in the subgroup of 100 patients who were exposed to the risk of developing rejection, with an overall rate of 89% during the first 3 months post-OLT, similar in the 3 groups. However, incidences of steroid-resistant rejection diagnosed during the 10 first days post-OLT were 54%, 24%, and 34% in groups I, II, and III and 46%, 26%, and 11%, respectively, during the 10-90 days interval. Sixteen patients with CMV had received OKT3, whereas the 5 remaining CMV cases had not (P = 0.019). In summary: (1) mAbs did not modify crude incidence of AR; (2) in the early period (< 10 days), TDIS immunoprophylaxis combined with OKT3 was more efficient than TDIS alone; (3) when compared with groups I and II, LO-Tact-1 apparently better prevented steroid-resistant rejection during the 10-90 days post-OLT; (4) OKT3 significantly increased incidence of CMV infection. In conclusion, TDIS with LO-Tact-1 seemed to achieve the better risk-benefit ratio in induction immunosuppression after OLT.

The aim of the study is to determine the effect degranulation of B cells on their immunohistochemical detection and to evaluate whether this effect depends on the technique of immunological detection. The biological model is the pancreatic insulin-containing B cell. To decrease the insulin content of the pancreas, insulin release was stimulated by five intraperitoneal injections of glibenclamide (2 mg/kg). Specimens of the pancreas were taken for insulin extraction and quantitation by radioimmunoassay (RIA), ultrastructural analyses and immunocytochemistry. The sections were treated either by polyclonal or monoclonal anti-insulin serum at various concentrations and peroxidase-antiperoxidase (PAP) technique eventually followed by silver amplification. The B-cell insular fraction or relative B-cell area (RBA) was measured with an automatic image analyser. The reference value for the relative B-cell area was established in control rats and corresponds to the ratio of the insular area occupied by immunostained B cells to the islet area. Its value was confirmed by calculation of the area occupied by non-B cells. RIA indicates a decrease of 83% of the insulin content in treated rats while the number of B granules decreases by 72% at the ultrastructural level. In usual conditions (polyclonal serum, 1/1500) the degranulation leads to a 16% underestimation of the B-cell insular fraction. This underestimation increases when monoclonal antibodies are used and further increases when higher dilutions are tested. The silver amplification does not prevent this underestimation and, in this particular model, exclusively acts by increasing the contrast. The only means of restoring the correct level of detection is to use the serum at a higher concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

A 67-year-old patient, with primary polymyositis and without previous evidence of liver disease, developed clinical and biochemical features of severe cholestasis 3 months after initiation of azathioprine therapy. Liver biopsy showed cholestasis with both cytological and architectural alterations of interlobular bile ducts. Azathioprine withdrawal resulted after 7 weeks in the resolution of clinical and biochemical abnormalities. It is believed that this is the first reported case of reversible azathioprine-induced cholestasis associated with histological evidence of bile duct injury.

Introduction Les sous-unites SUR1 et Kir6.2 contiennent dans leur sequence amino-acide un signal de retention qui controle leur association physique en un complexe hetero-octamerique formant les canaux KATP fonctionnels. Theoriquement, quand ces sous-unites ne sont pas co-exprimees ce signal de retention empeche leur migration vers la membrane cytoplasmique. Nous avons etudie le devenir de la sous-unite Kir6.2 en absence de SUR1 en comparant les cellules endocrines du pancreas de souris controle (WT) et de souris knockout pour le gene codant la sous-unite SUR1 (Sur1-/-). Materiels et methodes Les localisations sub-cellulaires de Kir6.2 et de la calreticuline (marqueur de reticulum) ont ete evaluees par des systemes de detections basees sur l’immunofluorescence en microscopie optique et sur l’immunogold en microscopie electronique. La longueur du reticulum rugueux lamellaire (L-RER) et la densite des particules d’or de Kir6.2 ont ete quantifiees par morphometrie. Resultats Comparee au WT, la surface de l’immunomarquage de la calreticuline est plus grande dans les cellules endocrines Sur1-/-, principalement autour des noyaux, ce qui reflete une plus grande densite du L-RER. L’immunolocalisation de Kir6.2 est superposable a celle de la calreticuline dans les 2 groupes et n’est pas observee a la membrane plasmique. En microscopie electronique, nous confirmons que le L-RER est plus abondant dans toutes les cellules endocrines chez les Sur1-/-, avec une longueur par surface de cellule endocrine 2 a 3x plus elevee, que chez les WT. La densite des particules d’or detectant Kir6.2 est aussi plus elevee dans le L-RER des cellules endocrines Sur1-/-refletant ainsi la sequestration de cette sous-unite dans cet organelle. Cependant, quelques particules d’or sont egalement observees dans les mitochondries et les granules de secretion. Conclusion En absence de SUR1, la majorite des sous-unites Kir6.2 est sequestree dans le L-RER mais une minorite echappe aux systemes de controle et se retrouve dans d’autres compartiments cellulaires en petite quantite.

We report the case of a patient with bone marrow transplantation in whom progressive cholestasis together with an extensive intrahepatic bile duct involvement, mimicking that usually observed in primary sclerosing cholangitis, occurred and was attributed to chronic GVHD

Relaxin has been localized in corpora lutea (CL) of pregnant NMRI mice using the avidin-biotin complex immunocytochemical procedure and an antiserum against highly purified porcine relaxin. The immunostaining was measured by immunodensitometry. Relaxin immunostaining was first observed in luteal cells of type I gestational CL on day 11.5 (D11.5). For each investigated day, all CL were identically stained, and immunostaining was evenly dispersed all over the CL. Seventy-five percent of cells were stained at D11.5, and nearly all cells were stained between D13.5 and D18.5. The staining intensity increased throughout the last half of pregnancy, reaching a maximum at D18. A few hours before parturition, at D18.5, relaxin immunostaining decreased dramatically and reached the background level shortly after delivery. From our results we may conclude that, in murine CL, the number of relaxin-secreting cells and the intracellular storage of the peptide increase during pregnancy. The disappearance of relaxin with the cells occurs rapidly +/- 12 h before parturition.

Of 139 children who received an orthotopic liver transplant in our center between March 1984 and July 1989, a total of 17 patients (12%) had transplants before their first birthday (mean age 10.3 months; range 8 to 11). The mean weight was 7.3 kg (range 5.2 to 13). Nine retransplantations were performed in five children because of primary nonfunction (three children), hepatic artery thrombosis (four), or rejection (two). A reduced donor liver was used for 11 of 26 transplants. Baseline immunosuppression included cyclosporine, prednisone, and azathioprine with OKT3 or anti-thymocyte globulin for steroid-resistant rejection episodes. Survivors were discharged after a mean hospital stay of 47 days (range 22 to 87), and nonsurvivors died within a mean of 40 days (range 0 to 120). The 1 year actuarial survival rate was 64.7%, in comparison with 75.8% in the whole series. One patient died perioperatively, two died from primary nonfunction, one from adenovirus infection, two from rejection, and one from bone marrow aplasia. Eighteen rejection episodes, of which 11 were steroid resistant, occurred in 11 patients. Our series shows that liver transplantation can be successful in this age group.

The exact nature of the beta-cell defect in type 2 diabetic patients is still unclear. beta-Cell mass reduction has been reported but remains controversial. A preliminary study of a large series of patients has demonstrated that in most, the beta-cell defect is not related to a decreased beta-cell mass. Amyloid deposits are observed in the islets of some type 2 diabetic patients but also in normoglycemic subjects. Because it has been claimed that these deposits interfere with beta-cell function, we evaluated in situ the effect of insular amyloid deposits on beta-cell transcription and translation. Pancreases were obtained at autopsy from 28 normoglycemic patients and 41 type 2 diabetic patients. Staining with hemaluneosin and Congo red was used to analyze the general features of the islets and the presence of amyloid deposits, respectively. Immunohistochemistry for proinsulin was performed with an antibody recognizing the junction between B-chain and C-peptide, thus specifically labeling the Golgi area where proinsulin is produced. In seven patients, we evaluated insulin gene transcription by in situ hybridization of proinsulin mRNA combined with Congo red staining, and we evaluated insulin storage by double immunostaining for insulin and amylin. In many type 2 diabetic patients, the islets appeared entirely normal. Amyloid deposits were found in 57% of diabetic subjects and 33% of normoglycemic age-matched control subjects. The percentage of amyloid-infiltrated islets varied from 0.4 to 74%. beta-Cells from amyloid-containing islets still had specific Golgi proinsulin labeling. In obese type 2 diabetic patients, the number of beta-cells with abnormal expression of proinsulin in the whole cytoplasm was significantly higher than in normoglycemic control subjects. Proinsulin mRNA was significantly reduced in islets with amyloid deposits when compared with amyloid-free islets, but the mean reduction did not exceed 16%. Insulin was still present in the beta-cells of amyloid-containing islets, and its amount, estimated by measurement of the insulin-labeling optical density, was not statistically different from that in amyloid-free islets. In conclusion, even in amyloid-containing islets, beta-cells maintain active insulin transcription and translation and normal insulin storage. Taking into account that in most cases only a small proportion of islets are infiltrated by amyloid, the limited reduction in proinsulin mRNA is unlikely to play a major role in the pathogenesis of diabetes.

Paternal mutation of ATP-sensitive K+ (KATP) channel genes and loss of heterozygosity (LOH) of the 11p15 region including the maternal alleles of ABCC8 , IGF2 , and CDKN1C characterize the focal form of persistent hyperinsulinemic hypoglycemia of infancy (FoPHHI). We aimed to understand the actual nature of FoPHHI in comparison with insulinoma. In FoPHHI, the lesion consists in clusters of β-cells surrounded by non–β-cells. Compared with adjacent islets, proinsulin mRNA is similar and proinsulin production higher ( P ≤ 0.02), indicating regulation at a translational level, with slightly lower insulin stock and lower ABCC8 peptide labeling ( P

Young infants are often refused for liver transplantation on the sole basis of young age. Over a total of 141 chidren who received an orthotopic liver transplantation in our centre (March 1984-July 1989), 17 patients (12%), were transplanted before they 1st birthday (15 biliary atresia, 1 Byler disease & 1 tyrosinemia). Mean age was 10,3 months (range 8-11) and mean weight 7,3 kg (range 5,2 - 13). A reduced liver was used 11 times over a total of 26 transplantations (42%). Immunosuppression included cyclosporine A, prednisone and azathioprine & OKT3 or ATG for steroid resistant rejection. Survivors were discharged after a mean hospital stay of 47 days (range 22-87) and the one year actuarial survival is 65% versus 77% in the whole serie. Reasons for retransplantation (29%) were primary non function (3), hepatic artery thrombosis (4) and rejection (2). 1 patient died perioperatively, 2 from primary non function, 1 from adenovirus infection, 2 from rejection & 1 from bone marrow aplasia. 18 rejection episodes, (11 corticoresistant) occured in 11 patients. Liver transplantation can be proposed to young infants without age limits being imposed.

Antibodies to different peptides produced from the gastrin precursor have been used in light microscopic- and electron microscopic-immunogold studies of gastrinoma tissue and normal antral mucosa. Antibodies to the C terminus of progastrin, which are known to react with the intact precursor, revealed immunoreactive material in the rough endoplasmic reticulum, Golgi region, and electron-dense granules in gastrinoma cells. In normal antrum these antibodies again revealed the Golgi region and a population of electron-dense granules. Other antibodies that react with the products of progastrin processing, but not the precursor, e.g., C-terminal and N-terminal gastrin 17 specific antibodies, revealed only granules. In addition to electron-dense granules already mentioned, the latter antibodies also revealed electron-lucent and intermediate granule populations, which in antrum were the major granule types. It is proposed that the intact precursor occurs in rough endoplasmic reticulum and Golgi; thereafter, in the immature electron-dense granules, and subsequently in electron-lucent granules, biosynthetic processing liberates gastrin 17 and gastrin 34, which are the major active products of gastrin gene expression.

In the present study, we investigated whether the analysis of cells and proteins collected by bronchoalveolar lavage (BAL) could accurately reflect the degree of functional impairment in pulmonary sarcoidosis. Eighteen patients with biopsy-proven sarcoidosis were prospectively evaluated. An inverse relationship was demonstrated between BAL coefficient of excretion relative to albumin (RCE) values of IgG and IgA and diffusion for carbon monoxide (Dco). A similar negative correlation existed with PaO2 at the end of a maximal exercise. Steroid therapy in five patients lowered concomitantly BAL RCE of IgA and IgG while Dco values increased. Immunoperoxidase studies in three lung biopsies revealed numerous Ig-containing cells within the lung parenchyma. We suggest that these BAL Ig values reflected the mononuclear cell infiltration of the bronchiolovascular sheaths and lung interstitium. This cellular infiltration likely induces a distortion of the capillary bed and may affect the gas exchanges in a reversible way.

The serum concentration of secretory component was monitored in 9 patients with massive liver necrosis. During acute cytolysis, no increase in serum secretory component levels was observed. Later on, patients with fulminant evolution displayed minor elevations. Values as high as those usually found in acute hepatitis were only observed in cases with delayed liver failure. In these patients, levels were elevated before failure, dropped as liver failure developed, and rose again during recovery. This evolution of serum secretory component level cannot be accounted for by a defective liver clearance of circulating secretory component. It rather suggests a release into the blood of secretory component produced in the liver by cells other than damaged hepatocytes, possibly during liver regeneration. In 3 cases, secretory component localization in the liver was demonstrated by immunocytochemistry and was compared with that in normal liver and in obstructive jaundice. In normal liver, it was restricted to portal bile duct cells and lumens. In obstructive jaundice, additional secretory component staining was found in bile canaliculi. After fatal liver necrosis, secretory component was localized to portal ducts and to a variable proportion of the cholangiocytelike cells belonging to the numerous extraportal proliferating ducts. In these structures, 0.1%, 21%, and 4% of the cells were stained 3, 8, and 30 days after maximal cytolysis, respectively. Remaining hepatocytes and bile canaliculi or bile plugs were unstained. Therefore, portal cholangiocytes and extraportal cholangiocytelike cells are probably essential sources of secretory component in human liver. We propose that proliferation of extraportal cholangiocytelike cells, expression of secretory component synthesis by these cells, and their inability to secrete secretory component in a disorganized biliary tree result in the elevated serum concentration of secretory component observed after acute liver necrosis.