Apmis

WILLEMOE, GRO LINNO; VAINER, BEN

doi: 10.1111/j.1600-0463.2009.02564.xpmid: 20132171

Willemoe GL, Vainer B. AMACR is not applicable as a diagnostic tool in hepatocellular carcinoma. APMIS 2010; 118: 85–90. α‐methylacyl coenzyme A racemase (AMACR or P504S) is a mitochondrial and peroxisomal protein present in a variety of human cells. Demonstration of increased expression is used diagnostically in prostatic adenocarcinoma. AMACR is also produced by normal hepatocytes and it has been postulated that the demonstration of AMACR expression or its pattern of distribution is useful in the diagnosis of hepatocellular carcinoma (HCC) (Jiang et al., Hum Pathol 2003;34, Guzman et al., Appl Immunohistochem Mol Morphol 2006;14, Li et al., J Exp Clin Cancer Res 2008;27). The aim of the present study was to evaluate whether immunohistochemical staining for AMACR can be used in a routine histopathologic setting. Immunohistochemical staining for AMACR was performed on paraffin‐embedded tissue from livers resected for HCC during 1980–2006 at Rigshospitalet, Copenhagen, Denmark (n = 44). Tumor sections as well as surrounding non‐neoplastic tissues were studied. In both tumor and non‐tumor tissues, intracellular localization and staining pattern were assessed and the staining intensity of AMACR was graded. The fraction of stained tumor cells was not significantly different from that of stained non‐tumor cells in the same patients (p = 0.97). A significantly lower staining intensity was observed in clear cell areas (p = 0.005), but the AMACR expression did not correlate with the HCC type and could not distinguish neoplastic from non‐neoplastic liver cells. AMACR is not applicable as a tool in the histopathologic diagnosis of HCC.

KAARTEENAHO, RIITTA; SORMUNEN, RAIJA; PÄÄKKÖ, PAAVO

doi: 10.1111/j.1600-0463.2009.02566.xpmid: 20132172

Kaarteenaho R, Sormunen R, Pääkkö P. Variable expression of tenascin‐C, osteopontin and fibronectin in inflammatory myofibroblastic tumour of the lung. APMIS 2010; 118: 91–100. The aim of this study was to analyse the expression of tenascin‐C, osteopontin and fibronectin in inflammatory myofibroblastic tumour of the lung, which is a rare tumour of unknown aetiology. Nine patients with an inflammatory myofibroblastic tumour of lung were studied by immunohistochemistry for the presence of tenascin‐C, osteopontin, fibronectin and alpha‐smooth muscle actin, which is a common marker for myofibroblasts. The ultrastructure of myofibroblasts was confirmed by transmission electron microscopy. The expression of tenascin‐C, osteopontin, fibronectin and alpha‐smooth muscle actin was also studied by immunoelectron microscopy. All cases displayed all of the studied extracellular matrix proteins and also alpha‐smooth muscle actin‐positive spindle‐shaped fibroblastic cells that were undoubtedly myofibroblasts. The immunoelectron microscopic studies demonstrated labelling for alpha‐smooth muscle actin in intracellular filament bundles within myofibroblasts, for fibronectin in the extracellular filaments of the fibronexus and for tenascin‐C extracellularly often adjacent to myofibroblasts. Labels for osteopontin were observed within myofibroblasts and plasma cells. These results demonstrate that tenascin‐C, osteopontin and fibronectin were expressed in all three kinds of subtypes of inflammatory myofibroblastic tumours of the lung and further, variable amounts of myofibroblasts could be observed by light and transmission electron microscopy as well as by immunoelectron microscopic techniques.

BRØCHNER, ANNE C.; DAGNÆS‐HANSEN, FREDERIK; TOFT, PALLE

doi: 10.1111/j.1600-0463.2009.02567.xpmid: 20132173

Brøchner AC, Dagnæs‐Hansen F, Toft P. Effect of renal and non‐renal ischemia/reperfusion on cell‐mediated immunity in organs and plasma. APMIS 2010; 118: 101–7. Acute renal failure (ARF) is a common morbidity factor among patients in the intensive care unit, reaching an incidence from 3% to 30% depending on the definition of ARF and the population. Although the majority of the patients with ARF are treated with continuous renal replacement therapy, the mortality rate still remains above 50%. The causes of death are primarily extra‐renal and include infection, shock, septicemia, and respiratory failure. We wanted to evaluate the cell‐mediated inflammatory response of renal ischemia–reperfusion (I/R) and non‐renal I/R, in blood and in distant organs. In our study, 80 mice were divided into four groups. The following surgeries were performed on the groups compared: bilateral renal I/R by clamping, unilateral renal ischemia, anesthesia only, and unilateral hind leg I/R. Half of the animals were killed after 2 h and the other half after 24 h. To assess the inflammatory response, we measured myeloperoxidase (MPO) in the organs, and CD 11b and major histocompatibility complex (MHC) II‐positive cells in the blood. Non‐renal I/R elicited the most elevated levels of MPO in extra‐renal tissue such as the lungs. There was a trend toward higher MPO levels in the kidney following renal I/R. All kinds of I/R induced an upregulation of the adhesion molecule CD 11b and a downregulation of MHC II. Renal and non‐renal I/R induced neutrophil infiltration in distant organs. Renal I/R does not induce a larger cell‐mediated inflammatory response in blood and organs than non‐renal I/R.

BOSSETO, MAIRA CEGATTI; PALMA, PATRICIA VIANNA BONINI; COVAS, DIMAS TADEU; GIORGIO, SELMA

doi: 10.1111/j.1600-0463.2009.02568.xpmid: 20132174

Bosseto MC, Palma PVB, Covas DT, Giorgio S. Hypoxia modulates phenotype, inflammatory response, and leishmanial infection of human dendritic cells. APMIS 2010; 118: 108–14. Development of hypoxic areas occurs during infectious and inflammatory processes and dendritic cells (DCs) are involved in both innate and adaptive immunity in diseased tissues. Our group previously reported that macrophages exposed to hypoxia were infected with the intracellular parasite Leishmania amazonensis, but showed reduced susceptibility to the parasite. This study shows that although hypoxia did not alter human DC viability, it significantly altered phenotypic and functional characteristics. The expression of CD1a, CD80, and CD86 was significantly reduced in DCs exposed to hypoxia, whereas CD11c, CD14, CD123, CD49 and HLA‐DR expression remained unaltered in DCs cultured in hypoxia or normoxia. DC secretion of IL‐12p70, the bioactive interleukin‐12 (IL‐12), a cytokine produced in response to inflammatory mediators, was enhanced under hypoxia. In addition, phagocytic activity (Leishmania uptake) was not impaired under hypoxia, although this microenviroment induced infected DCs to reduce parasite survival, consequently controlling the infection rate. All these data support the notion that a hypoxic microenvironment promotes selective pressure on DCs to assume a phenotype characterized by pro‐inflammatory and microbial activities in injured or inflamed tissues and contribute to the innate immune response.

ZAFIRELLIS, KYRIAKOS; ZACHAKI, AGLAIA; AGROGIANNIS, GEORGE; GRAVANI, KATERINA

doi: 10.1111/j.1600-0463.2009.02569.xpmid: 20132175

Zafirellis K, Zachaki A, Agrogiannis G, Gravani K. Inducible nitric oxide synthase expression and its prognostic significance in colorectal cancer. APMIS 2010; 118: 115–24. Nitric oxide synthases (NOS) are expressed in colorectal cancer. The aim of this study was to examine the inducible NOS (iNOS) expression in colorectal cancer and to investigate its prognostic relevance. Tissue sections of primary tumors from 132 patients undergoing curative resection for colorectal cancer were immunohistochemically examined for iNOS expression. The expression pattern of iNOS was correlated with various clinicopathological characteristics and survival. iNOS immunoreactivity was observed in the cytoplasm of tumor epithelial cells in 60 patients (45.5%) and positively correlated with lymph node involvement (p = 0.019). No significant correlation was found between iNOS expression and various clinicopathological characteristics, including age, gender, tumor location, tumor size, tumor grade, T stage, and Union International Contra la Cancrum (UICC) stage. Survival analysis showed a significant correlation between iNOS‐positive tumors and poor disease‐specific survival (p < 0.0001), with independent prognostic significance in multivariate analysis (HR = 4.42; p < 0.0001). Patients with stage II disease and iNOS‐positive tumors had significantly worse disease‐specific survival than those with iNOS‐negative tumors (p < 0.0001). In addition, patients with stage III disease and iNOS‐positive tumors had significantly worse disease‐specific survival than those with iNOS‐negative tumors (p = 0.001). The ability of iNOS to predict outcome in colorectal cancer patients may be independent of other known prognostic factors, providing a new molecular marker with significant potential for clinical utility.

SOUSA, MIREILLE ÂNGELA BERNARDES; MENDES, EDILBERTO NOGUEIRA; APOLÔNIO, ANA CAROLINA MORAIS; FARIAS, LUIZ DE MACÊDO; MAGALHÃES, PAULA PRAZERES

doi: 10.1111/j.1600-0463.2009.02570.xpmid: 20132176

Sousa MÂB, Mendes EN, Apolônio ACM, Farias LM, Magalhães PP. Bacteriocin production by Shigella sonnei isolated from faeces of children with acute diarrhoea. APMIS 2010; 118: 125–35. Shigella is a common agent of diarrhoea, a worldwide major health problem. The bacterium produces bacteriocins; however, the role of these substances as a virulence factor is completely unknown. With the aim to search for colicin production by Shigella sonnei, to evaluate the influence of culture conditions on bacteriocin expression, and to characterize the substance partially, 16 S. sonnei strains isolated from children with diarrhoea were tested for antagonism against members of the intestinal microbiota or agents of diarrhoea. Nine strains exhibited isoantagonism and heteroantagonism against S. flexneri and diarrhoeagenic Escherichia coli. Autoantagonism and antagonism against the intestinal microbiota were not detected. Culture medium and incubation conditions influenced antagonism expression. Antagonism resulting from bacteriophages, low pH, fatty acids, hydrogen peroxide, and chloroform was excluded. The activity of the intracellular fraction obtained with 75% ammonium sulphate was preserved at pH 1.0–11.0, and was found to be reduced by organic solvents and affected by high temperatures and proteases. The antagonistic spectrum and the in vitro conditions for better antagonism expression suggest that the role of colicin in S. sonnei virulence, if any, would be expressed prior to infection, and may regulate population density of enteropathogens by helping in organism transmission.

GRADEL, KIM O.; NØRGAARD, METTE; SCHØNHEYDER, HENRIK C.; DETHLEFSEN, CLAUS; EJLERTSEN, TOVE; KRISTENSEN, BRIAN; NIELSEN, HENRIK

doi: 10.1111/j.1600-0463.2009.02571.xpmid: 20132177

Gradel KO, Nørgaard M, Schønheyder HC, Dethlefsen C, Ejlertsen T, Kristensen B, Nielsen H. Salmonella or Campylobacter gastroenteritis prior to a cancer diagnosis does not aggravate the prognosis: a population‐based follow‐up study. APMIS 2010; 118: 136–42. We hypothesized that preceding zoonotic Salmonella or Campylobacter gastroenteritis aggravated the prognosis in cancer patients. Exposed patients comprised all of those diagnosed with first‐time Salmonella/Campylobacter gastroenteritis from 1991 and with first‐time cancer diagnosis thereafter (through 2003) in two Danish counties. These patients were matched for main cancer type, gender, age and calendar period to unexposed cancer patients, i.e. those without Salmonella/Campylobacter gastroenteritis. We compared cancer stage by age‐ and comorbidity‐adjusted logistic regression analysis, survival by comorbidity‐adjusted Cox’s regression analysis and mortality dependent on the time period between Salmonella/Campylobacter gastroenteritis and cancer by spline regression curves. The study cohort comprised 272 Salmonella/Campylobacter‐exposed cancer patients and 2681 unexposed cancer patients. Prevalence odds ratios (95% confidence intervals (CI)) in exposed as compared with unexposed patients were 0.96 (0.74–1.25) for localized tumours, 1.15 (0.87–1.54) for regional spread and 1.14 (0.84–1.55) for metastases. Adjusted mortality rate ratios (95% CI) were 0.93 (0.75–1.16) for 0–1 year, 1.08 (0.84–1.39) for 2–5 years and 1.02 (0.60–1.73) for the remaining period. Mortality estimates did not change in relation to the time period between gastroenteritis and cancer. Salmonella/Campylobacter gastroenteritis prior to cancer was associated with neither the cancer stage nor a poorer prognosis.

WISING, CATHARINA; MAGNUSSON, MARIA; AHLMAN, KARIN; LINDHOLM, LEIF; LAGERGÅRD, TERESA

doi: 10.1111/j.1600-0463.2009.02573.xpmid: 20132178

Wising C, Magnusson M, Ahlman K, Lindholm L, Lagergård T. Toxic activity of the CdtB component of Haemophilus ducreyi cytolethal distending toxin expressed from an adenovirus 5 vector. APMIS 2010; 118: 143–9. The Haemophilus ducreyi cytolethal distending toxin (HdCDT) catalytic subunit CdtB has DNase‐like activity and mediates DNA damage after its delivery into target cells. We constructed a replication‐deficient adenovirus type 5 (Ad5) vector expressing CdtB and investigated the toxic properties of this vector on HeLa cells. Ad5CdtB caused loss of cell viability, morphologic changes, and cell cycle arrest, findings similar to HdCDT intoxication. This confirmed that CdtB is responsible for the toxicity of the holotoxin when expressed in cells following transduction by an adenoviral vector, and indicated a possible potential of this novel strategy in studies of activity of intracellular products and in gene therapy of cancer.

CAKIR, EBRU; DEMIRAG, FUNDA; AYDIN, MEHTAP

doi: 10.1111/j.1600-0463.2009.02575.xpmid: 20132179

Cakir E, Demirag F, Aydin M. Cytopathologic differential diagnosis of small cell carcinoma and poorly differentiated non‐small cell carcinoma in bronchial lavage specimens using a regression analysis. APMIS 2010; 118: 150–55. The aim of this study was to determine the most significant cytologic features to differentiate small cell carcinoma (SCC) from poorly differentiated non‐small cell carcinoma (NSCC) in bronchial lavage specimens. Bronchial lavage specimens from 35 SCC cases and 63 poorly differentiated NSCC cases were examined and the cytologic parameters reviewed retrospectively. Thirty‐five cytologic features considered useful in differential diagnosis were assessed. Statistical analysis indicated that salt and pepper chromatin, small cell size and nuclear molding have more than 90% sensitivity and 70% specificity for SCC cases. Logistic regression analysis demonstrated that the most effective criteria to differentiate SCC from poorly differentiated NSCC are small cell size, salt and pepper chromatin, prominent nucleolus and papilla formation. When these selected variables were used, sensitivity for predicting SCC was 94.3% and specificity 96.8%, and sensitivity for predicting NSCC was 96.8% and specificity 94.3%. There are several cytologic features, which are highly sensitive and specific for distinguishing SCC from NSCC. Nuclear features such as chromatin pattern, and size of the nucleoli and nuclei are more valuable than cytoplasmic features to distinguish between the two.

WERTHÉN, MARIA; HENRIKSSON, LINA; JENSEN, PETER ØSTRUP; STERNBERG, CLAUS; GIVSKOV, MICHAEL; BJARNSHOLT, THOMAS

doi: 10.1111/j.1600-0463.2009.02580.xpmid: 20132180

Werthén M, Henriksson L, Jensen PØ, Sternberg C, Givskov M, Bjarnsholt T. An in vitro model of bacterial infections in wounds and other soft tissues. APMIS 2010; 118: 156–64. There is growing evidence that bacteria play a crucial role in the persistence of chronic wounds. These bacteria are most probably present in polymer‐embedded aggregates that represent the biofilm mode of growth. Much work has been carried out to study the development of biofilms in vitro, in particular in attachment to solid surfaces. The observations from the chronic wounds indicate that the bacteria are not attached to a solid surface. Consequently, a new in vitro model is required to investigate biofilms in more wound‐like settings. This study describes such a novel in vitro model, with bacteria growing as biofilm aggregates in a collagen gel matrix with serum protein mimicking the wound bed of chronic wounds. The model was verified to comprise important hallmarks of biofilms such as the bacterial embedment in a matrix and increased antibiotic tolerance. Furthermore, we have verified the relevance of the model by comparing the organization of the bacteria in the model with the organization of the bacteria in a real chronic wound. We believe that we have developed an important new model for investigating bacterial biofilms in chronic wounds. This model may be used to study biofilm development in chronic wounds and to develop novel diagnostic tools as well as treatment strategies.