The continuing search for Mycobacterium tuberculosis involvement in sarcoidosis: a study on archival biopsy specimens

Introduction: Increasing evidence indicates that mycobacteria may be involved in the aetiology and pathophysiology of sarcoidosis. Objectives: To investigate the association between Mycobacterium tuberculosis complex infection and sarcoidosis. Methods: Mediastinal lymph node biopsy specimens (formalin‐fixed, paraffin‐embedded) from 52 Danish patients with sarcoidosis, 50 patients with mediastinal lymphadenopathy of other non‐mycobacterial causes (negative controls) and 12 patients with histologically and/or culture‐verified mycobacteriosis (positive controls) were included in the study. Biopsy samples were analysed for the presence of Mycobacterium tuberculosis complex by strand displacement assay and a subset of specimens were examined for bacterial rRNA by fluorescent in situ hybridisation using an eubacterial probe with general bacterial specificity (EUB338). Results: One patient with sarcoidosis displayed a positive M. tuberculosis complex test. All negative controls were negative in the test and 5/12 patients with mycobacteriosis were positive in the test. We detected M. tuberculosis complex DNA in 10‐year‐old biopsy samples. Thirty‐six samples were tested with the eubacterial probe; of these, 67% were positive with no difference between patients and controls. Conclusion: Our results do not support the hypothesis that M. tuberculosis complex infection is involved in the pathogenesis of sarcoidosis. However, we stress the importance of excluding mycobacteriosis in the diagnostic workup of sarcoidosis patients. Please cite this paper as: Svendsen CB, Milman N, Rasmussen EM, Thomsen VØ, Andersen CB and Krogfelt KA. The continuing search for Mycobacterium tuberculosis involvement in sarcoidosis: a study on archival biopsy specimens. Clin Respir J 2011; 5: 99–104.     

Angiotensin converting enzyme (ACE) gene polymorphism in sarcoidosis in relation to associated autoimmune diseases

Abstract. Papadopoulos KI, Melander O, Orho‐Melander M, Groop LC, Carlsson M, Hallengren B (University of Lund, Malmö University Hospital, Malmö, Sweden). Angiotensin converting enzyme (ACE) gene polymorphism in sarcoidosis in relation to associated autoimmune diseases. J Intern Med 2000; 247: 71–77. Objectives. To investigate the significance of ACE gene insertion/deletion (I/D) polymorphism in the frequency of autoimmune manifestations in sarcoidosis. Design. In patients with sarcoidosis the ACE gene I/D polymorphism was detected with PCR on genomic DNA. The patients with sarcoidosis were divided according to the presence (n = 30) or absence (n = 32) of autoimmune manifestations. The former group was subdivided into thyroid autoimmunity (n = 10), gluten immune reactivity (n = 10) and gastric autoimmunity (n = 17). Settings. The patients were recruited at the Department of Pulmonary Medicine, and the study was conducted at the Department of Endocrinology, University of Lund, Malmö University Hospital, Malmö, Sweden. Subjects. Sixty‐two patients with documented sarcoidosis (30 females, 32 males, median age/range at diagnosis of sarcoidosis 31.5/19–75 years, median age/range at study 47.5/22–81 years) were examined. A total of 107 healthy unrelated subjects without sarcoidosis (60 females, 47 males, median age/range at study 58/40–82 years) served as controls. Results. S‐ACE values were significantly increased in patients compared to controls (P = 0.00001). The same was true in the subgroup of sarcoidosis patients with associated autoimmunity compared with those with isolated sarcoidosis (P = 0.0328). A significant association was seen between ACE gene polymorphism (II, ID, DD genotypes) and S‐ACE levels in both patients and controls according to the order II < ID < DD. The observed genotype frequency distributions in the different study groups agreed the Hardy–Weinberg equilibrium without significant differences between the patients and the controls. Within the group with autoimmune manifestations the DD genotype was significantly over‐represented in X‐ray stage III compared to the other X‐ray stages (P = 0.0181) and a significant increase in the DD genotype in X‐ray stage III (P = 0.035) in the group with autoimmune manifestations compared to isolated sarcoidosis was detected. Conclusion. We confirmed that the S‐ACE levels corresponded to the order II < ID < DD in patients with sarcoidosis as well as in healthy controls. S‐ACE levels were significantly higher in sarcoidosis patients with autoimmune manifestations. The frequency of the DD genotype was significantly increased in patients with autoimmune manifestations and major granuloma mass (X‐ray stage III). The ACE D allele in its homozygous form may confer susceptibility for autoimmune manifestations in sarcoidosis, possibly via the high levels of S‐ACE it encodes.     

Antimycobacterial immune responses in patients with pulmonary sarcoidosis

Introduction: Sarcoidosis is a multisystem granulomatous disease of unknown origin. Pathogenetic involvement of Mycobacterium tuberculosis has frequently been discussed in the aetiology of sarcoidosis; however, studies still remain contradictory. Objective: We addressed the question of mycobacterial involvement in the pathogenesis of sarcoidosis by analysing cellular immune responses to mycobacterial antigens. Methods: We examined the interferon (IFN)‐γ production by enzyme‐linked immunospot in response to purified protein derivate (PPD) mycobacterial‐specific antigen early secretory antigenic target (ESAT)‐6 and culture filtrate protein (CFP)‐10 by peripheral blood mononuclear cells (PBMCs) and bronchoalveolar‐lavage mononuclear cells (BALMCs) of patients with pulmonary sarcoidosis, smear‐negative tuberculosis and controls. Results: Release of IFN‐γ in response to ex vivo contact with PPD, ESAT‐6 or CFP‐10 by BALMC and PBMC were comparable among patients with sarcoidosis and controls (PBMC P = 0.2326; BALMC P = 0.1767) and were less frequently observed in both groups compared to patients with tuberculosis (BALMC P < 0.05; PBMC P < 0.0001). Within PBMC, the immunophenotype of sarcoidosis patients differed from that of patients with tuberculosis, as well as from that of controls, while within BALMC it resembled that of patients with tuberculosis. Conclusion: In contrast to patients with tuberculosis, the frequency of mycobacteria‐specific local and systemic immune responses is not elevated in patients with sarcoidosis when compared to controls. The immunophenotype represents the local resemblance of the granulomatous reaction underlying tuberculosis and sarcoidosis while showing systemical difference. These observations do not support a role of an infection with M. tuberculosis in the pathogenesis of sarcoidosis. Please cite this paper as: Hörster R, Kirsten D, Gaede KI, Jafari C, Strassburg A, Greinert U, Kalsdorf B, Ernst M and Lange C. Antimycobacterial immune responses in patients with pulmonary sarcoidosis. The Clinical Respiratory Journal 2009; 3: 229–238.     

Relationship between serum levels of macrophage migration inhibitory factor and the activity of antineutrophil cytoplasmic antibody-associated vasculitides

Macrophage migration inhibitory factor (MIF) is a central proinflammatory cytokine that regulates innate and adaptive immune responses. To evaluate its role in primary vasculitides, we determined MIF by enzyme-linked immunoassay in the sera of patients with Wegener's granulomatosis (WG; n=26), microscopic polyangiitis (MPA; n=10), polyarteritis nodosa (PAN; n=9) and giant cell arteritis (GCA; n=11). Healthy controls (n=26) and patients with sarcoidosis (n=14) were studied in parallel. Serum levels of MIF were significantly higher in patients with WG (median 41.1, range 3.2–120 ng/ml) than those in healthy controls (6.0, 0.015–36.5 ng/ml; P<0.001) and in patients with sarcoidosis (13.8, 0.015–67.1 ng/ml; P<0.05). MIF values were higher in MPA patients (29.5, 9.9–69.4 ng/ml; P<0.01) in comparison with those in healthy controls. In particular, increased levels of MIF were associated with active disease as assessed by the Birmingham Vasculitis Activity Score. Sequential studies showed decreased levels of MIF after initiation of immunosuppressive therapy, with clinical improvement in WG and MPA patients. In contrast, serum levels of MIF were not significantly elevated in patients with PAN and GCA. The results suggest that MIF contributes to the inflammatory process and correlates with disease activity in antineutrophil cytoplasmic antibody-associated vasculitides.     

The association of serum total sialic acid/total protein ratio with diabetic parameters in young type 1 diabetic patients

Abstract Sialic acid is a terminal component of the non-reducing end of carbohydrate chains of glycoproteins and glycolipids. The purpose of this study was to estimate serum total sialic acid (TSA) concentrations and serum TSA/serum total protein (TP) ratios in young type 1 diabetic subjects and to investigate their association with diabetes-related parameters in that population. Twentyfour young type 1 diabetic patients and 20 healthy controls were enrolled in this study. Serum TSA and serum TSA/TP ratio were measured in both groups. Moreover, we looked for correlation among serum TSA, serum TSA/TP ratio and clinically relevant parameters such as urinary albumin excretion, blood pressure, diabetes duration, HbA1c, daily insulin dose, serum lipids and magnesium in type 1 diabetic patients. Serum TSA concentrations and serum TSA/TP ratio showed no statistical difference between patients and controls (p>0.05). While serum TSA concentrations only correlated with urinary albumin excretion (r=0.44, p=0.028), serum TSA/TP ratio correlated with diastolic blood pressure (r=0.48, p=0.015), diabetes duration (r=0.46, p=0.022) and urinary albumin excretion (r=0.53, p=0.007) in the diabetic subjects. We concluded that serum TSA/TP ratio might be a better indicator than serum TSA as an index of diabetic complications.     

Serodiagnosis of tuberculosis: Detection of mycobacterial antibodies and antigens

Setting: The diagnosis of tuberculosis is based primarily on identification of mycobacteria and on clinical evidence. Recently, serological studies have been widely used experimentally as a diagnostic approach.Objective: The aim of our study was to optimize serodiagnosis of tuberculosis by detecting mycobacterial antigens and antibodies in sera from patients with lung tuberculosis, non-related diseases and healthy controls.Design: Mycobacterium tuberculosis H37Rv was disintegrated by pressure. Cell walls were extracted with 3 M KCL and were subjected to gel filtration in Toyopearl gel. Immune sera were prepared by immunization of rabbits with cell wall material. Anti H37Rv antibodies were purified by affinity chromatography. The reagents obtained were used to detect serum antibodies and antigens (following immune complex dissociation) using ELISA.Results: Using fraction 6 of cell wall extract, antibodies were detected in 72.2% of TB patients; there were no positive reactions in control subjects. By use of affinity-purified antibodies, antigens were detected in 77.1% of TB patients, 10% of patients with unrelated diseases and 6.7% of healthy controls.Conclusion: Effective serodiagnosis of tuberculosis can be achieved only by combining detection of both circulating antibodies and antigens using highly specific purified reagents and immune complex-dissociated sera.     

Th1 and Th17 immune responses to viable Propionibacterium acnes in patients with sarcoidosis

BackgroundPropionibacterium acnes and Mycobacterium tuberculosis have emerged as probable candidates responsible for sarcoidosis. This study was conducted to investigate the Th1/Th17 responses elicited by these pathogens in sarcoidosis and to clarify the causative role of these pathogens.MethodsPeripheral blood mononuclear cells (PBMCs) obtained from patients with sarcoidosis and from healthy volunteers were, respectively, co-cultured with viable P. acnes, with Bacille de Calmette et Guérin (BCG) as a viable M. tuberculosis complex, and with the early secretory antigenic target (ESAT)-6. Th1 cytokine production was measured using RT-PCR and enzyme-linked immunospot (ELISPOT) assays, and interleukin (IL)-17 mRNA expression was measured by RT-PCR.ResultsIL-2 secretion from PBMCs after stimulation with P. acnes was significantly higher in patients with sarcoidosis than in the controls. Similarly, IL-2 and IL-12 mRNA expression after stimulation with P. acnes was significantly higher in PBMCs from patients with sarcoidosis than in PBMCs from controls. In contrast, IL-17 mRNA expression was significantly lower in PBMCs from patients with sarcoidosis than in PBMCs from controls. No significant differences between the groups were observed in the responses to stimulation with BCG or ESAT-6.ConclusionSarcoidosis may arise from an imbalance of Th1/Th17 immune responses against viable P. acnes, but not M. tuberculosis complex.     

Esophageal pH and bile monitoring for detecting duodenogastroesophageal reflux and evaluating the effectiveness of treatment

OBJECTIVE: To study methods of diagnosing duo­denogastroesophageal reflux (DGER) and to evaluate the role of bile reflux in gastroesophageal reflux disease (GERD). METHODS: Simultaneous 24‐h esophageal mucosal bilirubin level and pH monitoring were performed in 20 healthy subjects and 52 patients with symptoms suggesting gastroesophageal reflux. Data were gathered by using an ambulatory duodenogastric reflux monitoring system (Bilitec 2000) and an ambulatory pH recorder (Digitrapper MKIII). An absorbance value of ≥0.14 was designated as the threshold value for the presence of bile reflux. Patients suffering from mixed pathological reflux of acid and bile were treated with hydrotalcite and cisapride for 4 weeks, then the monitoring was repeated. RESULTS: No pathological acid reflux was found in the 20 healthy subjects. Based on findings from 24‐h esophageal pH monitoring, 47 of 52 patients (including 12 with esophagitis) were found to have patho­logical reflux. The total fraction of time with bile reflux was greater in the patients (n = 52) than in the healthy subjects (n = 20; 2.67 ± 3.23%vs 0.47 ± 0.71%; P < 0.05) and was greater in the patients with esophagitis (n = 12) than in the patients without esophagitis (n = 35; 5.41 ± 4.93%vs 1.68 ± 1.76%; P < 0.05). Of the 47 patients with GERD, 15 (32%) had abnormal mixed acid and bile reflux. Eight of the 15 patients with esophagitis were found to have mixed reflux and only four of 32 had acid reflux only (P < 0.05). After treatment with hydrotalcite and cisapride, the total fraction of time with acid and bile reflux in the 15 patients with mixed reflux decreased significantly (P < 0.05). Esophagitis was cured in seven of eight patients. CONCLUSION: The ambulatory duodenogastric reflux monitoring system is a useful tool for detecting DGER and evaluating the efficacy of treatment. Approximately 32% (15/47 cases) of patients with GERD were found to have DGER. Bile reflux may play a significant role in causing esophageal mucosal damage in patients suffering from GERD.     

Studies of phagocytic and killing activities of alveolar macrophages in patients with sarcoidosis

Phagocytosis and killing activities of alveolar macrophages were compared in 17 patients with stage 1 sarcoidosis and 6 healthy controls. The average total cell count of bronchoalveolar lavage fluid from patients with sarcoidosis was 7.8 ± 7.5 × 10⁶ cells; 70.4 ± 15% of these cells were alveolar macrophages and 25.9 ± 16.2% lymphocytes. Average total cell count from controls was 8.33 ± 8.6 × 10⁶ cells, with 92.7 ± 5.9% alveolar macrophages and 6.6 ± 4.4% lymphocytes. Purified alveolar macrophages were tested in in vitro antibacterial assays using S. aureus as a test microbe. Moderate decreases in the kinetics of staphylococcal ingestion were detected in the sarcoidosis group. The intracellular killing activity of macrophages was much lower in the patients with sarcoid than in control subjects. In a pilot study, intracellular killing activity of macrophages from 1 patient: with sarcoidosis was greatly enhanced by 24 hr treatment with transfer factor. In summary, alveolar macrophages from patients with radiographic stage 1 sarcoidosis have decreased bacterial ingestion and intracellular killing activities. These results suggest that macrophages undergo complex functional changes in sarcoidosis that may influence both disease development and host defenses. Offprint requests to: P. Orosi     

Recovery of cell wall-deficient organisms from blood does not distinguish between patients with sarcoidosis and control subjects

STUDY OBJECTIVE: To determine if cell wall-deficient forms (CWDF) of mycobacteria can be grown in culture of blood from subjects with sarcoidosis. DESIGN: A special multicenter study of sarcoidosis (A Case Control Etiologic Study of Sarcoidosis), supported by the National Heart, Lung, and Blood Institute. Patients and control subjects: Patients and control subjects were recruited at 10 institutions in the United States. Control subjects (controls) were of the same gender and race, and within 5 years of age as matching patients with sarcoidosis (cases). RESULTS: Cultures were incubated from 347 blood specimens (197 cases, 150 controls). Two investigators trained to recognize CWDF mycobacteria examined material obtained from culture tubes after 3 weeks. Structures thought to be CWDF were seen with equal frequency in cases (38%) and controls (41%). Thirty-nine percent of cases and 37% of controls were read as negative for CWDF. CONCLUSION: This study fails to confirm earlier reports that CWDF mycobacteria can be grown from the blood of patients with sarcoidosis, but not from control subjects.     

The BTNL2 A allele variant is frequent in Danish patients with sarcoidosis

Background: The butyrophilin‐like 2 (BTNL2) gene is located on chromosome 6p21.3 close to the HLA‐class II genes. An association has been reported between sarcoidosis and a single nucleotide polymorphism in BTNL2, rs2076530, also termed the A allele. Objectives: To evaluate whether patients with sarcoidosis carry the A allele more frequently than healthy subjects. Methods: The series comprised 87 ethnic Danes with sarcoidosis and 113 healthy control subjects. Analysis of rs2076530 was performed by Taqman assay, polymerase chain reaction and sequencing of genomic DNA. Results: Sarcoidosis patients had a higher frequency of the A allele than controls (73.9% vs 55.8%) (P < 0.025). The frequency of GG, GA and AA genotype was 5.7%, 40.2% and 54.0% in patients vs 16.0%, 56.6% and 27.4% in controls (P < 0.001). The AA genotype was associated with increased risk of sarcoidosis in both a dominant [odds ratio (OR) 3.1; 95% confidence interval (CI) 1.1–8.7; P < 0.03] and a recessive model (OR 3.1; 95% CI 1.72–5.61; P < 0.001). Population attributable fraction for disease was 50% in a dominant model and 25% in a recessive model. Conclusions: The BTNL2 A allele variant occurs with a high frequency in Danish patients with sarcoidosis and the AA genotype is associated with a ～threefold higher risk of sarcoidosis than the GG genotype. Our results should encourage future studies on the interrelationship between the BTNL2 protein and granuloma formation in sarcoidosis. Please cite this paper as: Milman N, Svendsen CB, Nielsen FC and Hansen TvO. The BTNL2 A allele variant is frequent in Danish patients with sarcoidosis. Clin Respir J 2011; 5: 105–111.     

Effects of pulse methylprednisolone and oral methylprednisolone treatments on serum levels of oxidative stress markers in Graves' ophthalmopathy

Background Recent evidence has shown that oxidative stress may play a role in the pathogenesis of autoimmune diseases, and this is an issue of considerable research interest in the field of infiltrative ophthalmopathy. Therefore, we evaluated both the relationship between Graves’ ophthalmopathy (GO) and serum levels of certain indicators of oxidative stress, and the effects of methylprednisolone treatment on serum malondialdehyde (MDA) and glutathione (GSH) levels in patients with euthyroid GO. Materials and Methods We compared GO patients to both Graves’ patients without ophthalmopathy and healthy controls. Ultimately, we assessed four subject groups. Graves’ patients with ophthalmopathy (GO) were subcategorized into two groups: Group A subjects (n = 18) were given intravenous glucocorticoid and Group B patients (n = 15) were given oral glucocorticoid. Graves’ patients without ophthalmopathy comprised Group C (n = 20), and healthy controls comprised Group D (n = 15). Serum levels of MDA and GSH were measured at baseline and after 4 and 24 weeks of observation via spectrophotometric methods. Results We found that serum MDA levels were significantly higher in the two GO groups (Groups A and B) than in GO patients without ophthalmopathy or healthy controls. Conversely, GSH levels were significantly lower in the two GO groups than in Groups C and D. MDA and GSH levels were not different between the latter two groups. MDA levels were strongly and positively correlated with a clinical activity score (CAS). In Group A, MDA levels and the CAS were significantly lower than in Group B at 4 weeks. After 24 weeks, however, MDA levels and the CAS were similar in these two groups. Conclusion Oxidative stress appears to be involved in the pathophysiology of GO. Relative to oral dosing, the intravenous administration of a glucocorticoid seems to yield more rapid improvement in disease activity. MDA might be useful as an indicator of clinical activity.     

Elevated homocysteine levels in patients with Raynaud's phenomenon secondary to systemic lupus erythematosus

The objective of this study was to analyse the association between serum homocysteine (Hcy) levels and the presence of Raynaud’s phenomenon (RP) in a cohort of systemic lupus erythematosus (SLE) patients. We enrolled premenopausal, disease-inactive SLE patients (n= 34) with RP (group I, n= 11) or without RP (group II, n= 23), and age-matched healthy premenopausal women as controls (group III, n= 20). Fasting Hcy levels were determined for all these subjects. The results reveal that group I patients exhibited significantly greater serum Hcy levels (11.68+/–2.98 mmol/l) than group II patients (8.29+/–2.89 mmol/l) (P= 0.003), and group III healthy subjects (8.00+/–1.50 mmol/l) (P= 0.001); however, in this regard there was no significant difference between group II and group III patients (P= 0.927). In conclusion, elevated serum Hcy levels were noted for this cohort of SLE patients with RP compared to those not evidencing RP and healthy controls. Although the pathogenesis of RP still remains obscure, this study suggests that Hcy may play a role in the aetiopathogenesis of SLE patients with RP. Received: 23 May 2001 / Accepted: 22 November 2001     

Serum B-cell maturation antigen is elevated in multiple myeloma and correlates with disease status and survival

Although TNFRSF17 (also designated as B‐cell maturation antigen (BCMA)) is expressed on tumour cells in B‐cell malignancies, it has not been found in serum. The present study found that BCMA concentrations were higher in the supernatants of cultured bone marrow mononuclear cells from multiple myeloma (MM) patients than in healthy subjects. Serum BCMA levels were measured in samples from MM patients (n = 209), monoclonal gammopathy of undetermined significance (MGUS) individuals (n = 23) and age‐matched controls (n = 40). BCMA was detected in the serum of untreated MM patients (n = 50) and levels were higher than in MGUS patients (P = 0·0157) and healthy subjects (P < 0·0001). Serum BCMA levels were higher among patients with progressive disease (n = 80) compared to those with responsive disease (n = 79; P = 0·0038). Among all MM patients, overall survival was shorter among patients whose serum BCMA levels were above the median (P = 0·001). We also demonstrated that sera from mice with human MM xenografts contained human BCMA, and levels correlated with the change in tumour volume in response to melphalan or cyclophosphamide with bortezomib. These results suggest that serum BCMA levels may be a new biomarker for monitoring disease status and overall survival of MM patients.     

Serum sialic acid in subjects with impaired glucose tolerance and in newly diagnosed type 2 diabetic patients

Several general population studies and those carried out in diabetic patients with complications have pointed to serum sialic acid as a marker of inflammation in atherosclerosis. In this study we examined whether total sialic acid (TSA) was changed in the sera of 28 newly diagnosed subjects with type 2 diabetes (type 2 DM), 47 subjects with impaired glucose tolerance (IGT) and 72 subjects with normal glucose tolerance (NGT). The associations between sialic acid and other atherosclerotic risk factors such as lipid profile, baseline diene conjugates in low-density lipoproteins (LDL-BDC) and fasting insulin were also investigated. We found a trend to TSA increase in subjects with impaired glucose tolerance and a significant increase in TSA in newly diagnosed patients with type 2 DM (2.2±0.3 vs. 1.9±0.3 mmol/l; p<0.03) when compared to subjects with NGT. Lipid profile and LDL-BDC, as a marker of circulating oxidized LDL, did not differ among glucose tolerance categories. Significant associations between total sialic acid and 2-h post-load glucose level, fasting insulin, insulin sensitivity, HDL-cholesterol and log of triglycerides were found in the examined subjects. Multiple regression analysis showed significant correlations between serum sialic acid and 2-h post-load glucose levels and insulin sensitivity. This study indicates that measurement of TSA as a marker of subclinical inflammation may be valuable as an independent parameter in identifying subjects at higher risk of developing type 2 diabetes and those who might benefit from anti-inflammatory treatment. Received: 10 May 2002 / Accepted in revised form: 15 April 2003 Correspondence to M. Gavella     

Hepatic granulomas: histological and molecular pathological approach to differential diagnosis–a study of 442 cases

Background/Aims: The incidence of hepatic granulomas is reported in 2–15% of liver biopsies. This study was carried out to evaluate the incidence and aetiology of hepatic granulomas in a German Institute of Pathology with specialization in liver diseases. Methods: A retrospective case review was performed on 12 161 liver biopsies of the Institute of Pathology (University of Cologne) between 1996 and 2004. Aetiology was determined according to histomorphological changes, clinicopathological data and liver tissue polymerase chain reaction (PCR) for detection of diverse putative pathogens in the liver tissue. Results: Four hundred and forty‐two liver biopsies revealed granulomatous lesions (3.63%). Two hundred and fifteen cases (1.77% of all biopsies and 48.64% of granulomatous lesions) were diagnosed as primary biliary cirrhosis. In 37 cases (0.3% of all biopsies and 8.37% of granulomatous lesions), the diagnosis of sarcoidosis was established. A positive PCR result for an infectious pathogen was obtained in 15 samples (3.39%) [Bartonella henselae (n=2), Listeria (n=3), Mycobacterium tuberculosis (n=3), Yersinia pseudotuberculosis (n=1), cytomegalovirus (n=2), Epstein–Barr virus (n=4)]. In six cases, a putative diagnosis was established according to the report of clinical conditions. In 11 cases (2.48%), drugs were the putative causative agent. In 158 cases (36%) a definite diagnosis could not be established. Conclusions: Hepatic granulomas have a broad range of underlying aetiologies. With a combined histological, clinical, serological, and molecular approach, we were able to clarify the cause in 64% of the cases. Owing to the diverse prognosis and therapeutic implications, a detailed interdisciplinary workup of all liver biopsies with granulomatous lesions is mandatory.     

A Comparison of Cognitive Function in Patients under Maintenance Treatment with Heroin,Methadone, or Buprenorphine and Healthy Controls: An Open Pilot Study

Introduction: Cognitive impairment has been reported in drug-dependent patients under opioid maintenance treatment. Objectives: To compare cognitive functioning in healthy controls and in opioid-dependent patients treated with Buprenorphine, Heroin, or methadone maintenance. Methods: We used the standardized test battery ART-90 to study cognitive function in patients under long-term heroin treatment (n = 20), Bup (n = 22), or Met (n = 24) maintenance treatment and healthy controls (n = 25). Results: Patients receiving heroin performed significantly worse than healthy controls in most domains. Heroin patients performed worse than patients in the other two treatment groups in subtests measuring psychomotor performance under stress conditions and monotony. Conclusions and Scientific Significance: Although a number of limitations must be taken into account, this study provides some preliminary evidence that cognitive function may be more impaired in patients under heroin maintenance treatment than in patients receiving Bup or Met and in healthy controls.     

The ability of anti-carbonic anhydrase II antibody to distinguish autoimmune cholangitis from primary biliary cirrhosis in Japanese patients

Serum antibody against carbonic anhydrase (CA) II has been described as a serological marker for distinguishing autoimmune cholangitis (AIC) from primary biliary cirrhosis (PBC). To validate this finding in a Japanese population, we evaluated sera from patients with PBC and AIC for antibody to human CA II. An enzyme-linked immunosorbent assay was employed to quantify serum antibody against CA II in patients with PBC (n = 40), AIC (n = 23), autoimmune hepatitis (n = 10), and extrahepatic obstructive jaundice (n = 10). Compared with the finding of a 4% prevalence of anti-CAII antibody in healthy subjects (n = 24), a significantly higher prevalence of anti-CA II antibody was detected in patients with PBC (35%) and AIC (30%) (P < 0.05), but not in patients with autoimmune hepatitis and patients with obstructive jaundice. No significant difference was observed between PBC and AIC patients. These results showed that AIC and PBC would be indistinguishable by anti-CA II antibody testing in Japanese patients. However, the finding of serum anti-CA II antibody in patients with PBC and AIC supports the disease concept of autoimmune exocrinopathy. Received: July 13, 1998/Accepted: October 23, 1998     

Slide agglutination test for the diagnosis of pulmonary and extra-pulmonary tuberculosis

Objective: Evaluation of a 2 min slide agglutination test to detect the presence of antibodies directed against Mycobacterium tuberculosis antigens.Design: The test utilizes solible antigens extracted from a cultivable new species of non-pathogenic saprophytic mycobacterium, Mycobacterium w, which shares antigenic determinants with M. tuberculosis. The soluble antigens are covalently linked to carboxylated polystyrene latex beads.Results: The sensitivity of the assay was increased from 78% (reported earlier), to 90.2% for pulmonary tuberculosis and 85.7% for extra-pulmonary tuberculosis. The specificity of the test was determined by testing the sera of apparently healthy controls, and patients with other respiratory tract infections and rheumatoid arthritis. Among the apparently healthy controls, 7.3% tested positive. None of the sera from the patients with other diseases gave positive agglutination.Conclusion: This simple and rapid technique could be suitable for mass screening for pulmonary and extra-pulmonary tuberculosis.     

CARD15 single nucleotide polymorphisms 8, 12 and 13 are not increased in ethnic Danes with sarcoidosis

BACKGROUND: Mutations of the caspase-activating recruitment domain 15 (CARD15) gene on chromosome 16 are associated with chronic inflammatory granulomatous bowel disease (Crohn's disease). Sarcoidosis is a systemic granulomatous disease with unknown etiology, which shares histological features with Crohn's disease. OBJECTIVES: To evaluate whether ethnic Danes with sarcoidosis have an increased frequency of CARD15 mutations compared to healthy control subjects. METHODS: Genotyping for CARD15 mutations R702W, G908R, and L1007fsinsC, also designated single nucleotide polymorphism (SNP) SNP8, SNP12 and SNP13, respectively, were performed by capillary electrophoresis single-strand confirmation polymorphism in 53 patients with histologically verified sarcoidosis and in 103 healthy controls. RESULTS: The frequencies of CARD15 mutations in sarcoidosis patients were: SNP8, 4/106 chromosomes (3.8%); SNP12, 2/106 chromosomes (1.9%); SNP13, 2/106 chromosomes (1.9%); SNP8+SNP12+SNP13, 8/106 chromosomes (7.6%). All 8 patients were heterozygous. The frequencies in controls were: SNP8, 9/206 chromosomes (4.4%); SNP12, 2/206 chromosomes (1.0%); SNP13, 4/206 chromosomes (1.9%); SNP8+SNP12+SNP13, 15/206 chromosomes (7.3%). All controls were heterozygous. The differences were not statistically significant (p>0.05). Furthermore, the course of disease was not significantly different in the 8 patients with CARD15 mutations and the 45 patients without mutations. CONCLUSION: The frequency of CARD15 mutations is not increased in ethnic Danish patients with sarcoidosis, and heterozygosity for such mutations apparently has no influence on the course of disease.