bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunohistochemical characterization of the cutaneous cellular infiltrate in different areas of chronic leg ulcers
Authors: Rosner K. , Ross C. , Karlsmark T. , Petersen A. A. , Gottrup F. , Vejlsgaard G. LANGE
Published: August 2009

Current understanding of the immunological mechanisms involved in the pathogenesis of venous leg ulcers is insufficient. In this study the cellular composition of skin biopsies taken from the center, the edge, and 2 cm distant from the edge of venous leg ulcers was characterized quantitatively by immunohistochemical staining. In the epidermis the mean numbers of Langerhans cells (CD1a+) were four times lower at the edge of the ulcer compared to clinically intact epidermis 2 cm distant from the edge. In the dermis a statistically significant increase in the mean numbers of macrophages (CD68+) and neutrophils (NP57+) from the distant area towards the center of the ulcer was observed. No significant differences were observed in the distribution of T cells nor in the ratio of CD4+/CD8+ T‐cell subsets between the different regions of the ulcer. About 30% of T lymphocytes were CD8+ in all microenvironments. The center and the edge of the ulcer were dominated by macrophages comprising 63% and 53% of the cells respectively, while T lymphocytes dominated the distant area. The area 2 cm distant from the edge was also heavily infiltrated by macrophages and neutrophils. B cells (CD22+) and NK cells (CD56+) were relatively rare in all areas, comprising less than 3% of the dermal infiltrate. In conclusion, local microenvironments each with a different cellular composition can be defined within venous leg ulcers.
Sample preparation for Chlamydia pneumoniae PCR
Authors: Gnarpe JUDY , Eriksson KARIN
Published: August 2009

Fifty‐three clinical specimens taken from the retropharyngeal mucosa of patients with longstanding respiratory tract infections were analyzed by polymerase chain reaction using two different methods for sample preparation. All specimens were divided into two aliquots, one treated with proteinase K, and the other with the Amplicor sputum sample preparation kit. All tests were run in parallel, employing a primer pair specific for Chlamydia pneumoniae. Of the samples prepared with the Amplicor kit 20.8% were found to be positive, as compared to 7.5% of the samples prepared with proteinase K. The outcome of C. pneumoniae PCR was improved by treatment involving a more complete lysis of cells derived from the specimen.
Human papillomavirus DNA and abnormal p53 expression in carcinoma of the urinary bladder
Authors: Kamel DIA , PÄÄKkÖ PAAVO , PÖLlÄNen RAIMO , VÄHÄKangas KIRSI , Lehto VELI‐PEKKA , Soini YLERMI
Published: August 2009

In this study we analysed 47 bladder carcinomas for the presence of DNA‐HPV subtypes 6, 11, 16, 18, 31 and 33 by nucleic acid in situ hybridization, and for the abnormal accumulation of p53 protein by immunohistochemistry. HPV DNA was found in 27/47 (57%) bladder carcinomas, with multiple subtypes in 20 cases. In squamous cell carcinoma (SCC), HPV DNA was only detected in the superficial layer of the neoplastic epithelium and was found mainly in the nuclear compartment. In contrast, in transitional cell carcinoma (TCC), HPV DNA was also found in deeper parts of the tumour. In about half the cases it was mainly found in the cytoplasmic compartment. In SCC, the HPV DNA labelling occurred in koilocytic cells, while no such association was found in TCC. Abnormal accumulation of p53 protein was found in 24/47 (51%) carcinomas. p53 positivity was found significantly more often in SCC than in TCC (p=0.05). Concurrent HPV positivity and abnormal p53 protein accumulation was found in 18 cases, 14 showing the presence of HPV subtypes 16 and/or 18 DNA. The results demonstrate that HPV DNA occurs widely in urinary tract tumours. Unlike in some other carcinomas, there was no inverse relationship between HPV positivity and abnormal p53 protein accumulation in bladder carcinomas. Thus HPV infection may play a role in the pathogenesis of bladder carcinomas by some mechanism other than inactivation of the p53 protein.
Development of a flow method for susceptibility testing of oral biofilms in vitro
Authors: Larsen TOVE , Fiehn NILS‐ERIK
Published: August 2009

Bacteria in biofilms are known to be more resistant than bacteria in batch cultures to antimicrobial agents. The purpose of the present study was to develop a flow method for formation of oral biofilms permitting susceptibility testing of plaque bacteria. A brain heart infusion (BHI) Streptococcus sanguis 804 culture was pumped through a modified Robbins Device (MRD) with 25 exchangeable silicone disks at 40 ml/h. After 24–48 h disks were removed and biofilm cells dispersed by vortex mixing and low‐output ultrasonication. Colony forming units (cfu)/cm2 were determined after aerobic incubation on blood agar plates. Optimal biofilm formation was found after growth for 48 h at 37°C in BHI + 1% sucrose, using saliva‐coated silicone disks in inverted MRDs, yielding on average 4.4 × 105 cfu/cm2. Similar results were obtained for S. sanguis ATCC 10556 and five clinical isolates. Testing the susceptibility of S. sanguis to chlorhexidine gluconate showed increased resistance of biofilms compared to batch culture. Thus an appropriate biofilm model for susceptibility testing of oral microorganisms has been established.
Experimental chronic Pseudomonas aeruginosa lung infection in rats
Authors: Lange KAI H. W. , Hougen HANS PETTER , HØIby NIELS , Fomsgaard ANDERS , Rygaard JØRGEN , Johansen HELLE KROGH
Published: August 2009

In a rat model of chronic Pseudomonas aeruginosa lung infection mimicking cystic fibrosis, we investigated the possibility of preventing chronic lung inflammation or decreasing the progression of the infection. We compared the lethality, pathology, bacterial clearance, and immunogenicity after stimulation of the non‐specific defence mechanisms by Escherichia coli lipopolysaccharide (LPS) or P. aeruginosa sonicate, or the acquired specific immune response by vaccination with the same bacterial antigens. One day prior to challenge with P. aeruginosa embedded in alginate beads, rats were stimulated with either E. coli LPS or P. aeruginosa sonicate. Four and two weeks prior to challenge other rats were vaccinated with either E. coli LPS or P. aeruginosa sonicate. Controls did not receive any stimulation or vaccination. The lethality after challenge was lower in rats stimulated with E. coli LPS (p= 0.02) or vaccinated with P. aeruginosa sonicate (p=0.03) as compared to controls. The histopathology of the surviving rats showed an acute inflammation dominated by polymorphonuclear leukocytes (PMNs), but the offending bacteria were not completely eliminated in any group. The increased survival was probably due to earlier recruitment of PMNs most likely mediated by either cytokines and other chemotactic factors (stimulated group) or the immune response in concert with the complement cascade (vaccinated group). The results of the present and previous vaccination studies show that it is possible to improve survival but not to prevent the chronic P. aeruginosa lung infection and inflammation caused by alginate‐embedded bacteria.
Head and neck cancer in Norway
Authors: Mork JON , Thoresen STEINAR , Faye‐Lund HILDE , Langmark FRØYDIS , Glattre EYSTEIN
Published: August 2009

Data from population‐based cancer registries provide information on the causes and outcome of cancer and form a basis for important decisionmaking in connection with the prevention of cancer and the planning of health services. This makes it of the utmost importance to assess the data at all stages of collection to ensure the highest possible quality. The present study focuses on the quality of the Cancer Registry of Norway's data on head and neck cancer for the period 1953–1991. When the study was started, 16,104 cases of head and neck malignancies had been registered. All histological codes were reviewed. The pathologists' reports were reevaluated for 369 cases selected according to set criteria: 133 cases received a new histological code without being exluded from the data material; 112 cases were excluded. The distribution of histological diagnoses for each location is presented. A reevaluation of 300 cases selected at random from the corrected series indicates discrepancies between the pathologist's classification and the Registry's coding in less than 2% (1.4%) of all cases. The percentage that lacked histological verification fell from 5.7% in the first decade to 2.1% during the last 9‐year period. Completeness of the Cancer Registry's data base was checked against hospital‐based registries and this investigation showed that virtually all new cases are reported. We conclude that the data on head and neck cancer for the studied time period meet standards that justify their use as a basis for epidemiological as well as clinical studies.
Evaluation of staphylococci isolated from a blood culture system (colorbact®)
Authors: JarlØV JENS OTTO , Prag JØRGEN , Rosdahl VIBEKE THAMDRUP , Espersen FRANK
Published: August 2009

During a one‐year period, all blood cultures positive for staphylococci from two of the smaller Danish counties with non‐university hospitals only were evaluated. The isolates were speciated and coagulase‐negative staphylococci (CoNS) were biotyped. Furtermore, antibiogram, phage typing and lectin typing were performed for all isolates. Clinical information was obtained by telephone, and before any bacteriological identification was performed, a preliminary judgment was made as to whether the positive blood culture was of clinical significance. A total of 3,500 blood cultures were evaluated and 426 (12.2%) were positive. One hundred and sixty blood cultures from 137 patients contained staphylococci; 36 of these patients had a pure culture of Staphylococcus aureus. One hundred and twenty‐four CoNS were found and identified as S. epidermidis (81), S. hominis (19), S. haemolyticus (8), S. simulans (1), and Micrococcus species (3), and another 12 staphylococcal isolates which could not be identified to species level. A total of 35 patients had mixed cultures, including 6 with S. aureus as one of the isolates and 15 with mixed CoNS. Clinical relevance was estimated in 90% of cases where the later bacterial identification showed 5. aureus, whereas clinical relevance was absent in the majority of cases including CoNS. Methicillin and gentamicin resistance was absent among S. aureus, but frequent among CoNS, especially S. epidermidis, where 40% were resistant to methicillin and 30% to gentamicin.
Immunogenicity expressed in patients with bacteraemia of an epitope shared by enterobacterial and neisserial porin proteins
Authors: Henriksen ARNE Z. , MæLand JOHAN A.
Published: August 2009

A monoclonal antibody (MAb) against an epitope (Po I) on an Escherichia coli O55 porin protein has shown broad cross‐reactivity with other Enterobacteriaceae and with both pathogenic and nonpathogenic Neisseriaceae. In this study, we have measured antibody levels against the Po I site in patients with bacteraemia in order to examine the immunogenicity of the Po I domain in humans. A MAb‐based competition ELISA (cELISA) was used. Only 20% of healthy controls had detectable levels of anti‐Po I antibodies in serum. Of patients bacteraemic with enterobacteria (n=45), 11% and 58% showed elevated antibody levels compared to healthy controls with the first and second serum specimens, respectively, and 73% of these patients showed ≥10% increase in the antibody levels. Of patients bacteraemic with N. meningitidis (n=20), only 30% showed ≥10% increase in the antibody levels when paired serum specimens were tested. Levels of competing antibodies were similar in the cELISA with N. meningitidis (B: 15: PI, 7, 16) OM coat or E. coli 055 OM coat. The results demonstrated that the highly conserved porin protein domain Po I expressed immunogenicity in humans when present in bacteria which caused bacteraemia. This finding represents a challenge in further investigations on the immunobiological role of the cross‐reacting antibodies.
Impaired function of postoperative macrophages from zinc‐deficient rats decreases collagen contraction
Authors: Ghassemifar M. REZA , Olsson MIKAEL G. , ÅGren MAGNUS S. , FranzÉN LENNART E.
Published: August 2009

Zinc deficiency impairs connective tissue contraction in the perforated rat mesentery model. Since the rat mesentery is almost avascular, free peritoneal macrophages are important for mesenteric repair. Impairment of contraction may thus be caused either by a direct effect of zinc deficiency on tissue cells or by hampered macrophage function. To further elucidate the role of macrophages in tissue contraction, we studied their effect on lattice contraction. A number of typical functions of macrophages in zinc deficiency were also investigated. Lattice contraction was significantly impaired by conditioned medium from zinc‐deficient macrophages. Zinc deficiency did not influence peripheral blood leukocyte number, but postoperatively the number of peritoneal macrophages increased on days 7 and 10. A significant release of lysosomal enzymes from macrophages was recorded during phagocytosis, whilst no difference was observed between controls and zinc‐deficient macrophages. Superoxide anion generation during phagocytosis was not significantly increased in zinc deficiency. Conditioned medium from zinc‐deficient macrophages was shown to impair lattice contraction in vitro and the results are compatible with impaired macrophage function as a cause of decreased connective tissue contraction in vivo.
Some new aspects of molecular mechanisms of cyclosporin A effect on immune response
Authors: Zav'Yalov VLADIMIR P. , Denesyuk ALEXANDER I. , Lundell JUHANI , Korpela TIMO
Published: August 2009

A few protein targets were found to display a specific high‐affinity interaction with the immunosuppressant cyclosporin A (CsA): cytosolic cyclophilins (CyP)A, B, C, D, E containing from 122 to 174 amino acid residues in a polypeptide chain, and secreted forms of CyP; CyP‐40,40‐kDa CsA‐binding polypeptide complexed with steroid receptor (SR); CyP‐related 150‐kDa receptor of natural killer (NK) cells; interleukin 8 (IL‐8); actin; a family of molecular chaperones hsp70 and P‐glycoprotein (P‐GP). All CyPs possess peptidyl‐prolylcis‐transisomerase activity (PPIase) and may serve as ATP‐independent molecular chaperone proteins. The CsA‐CyP complexes are specific inhibitors of Ca2+‐ and calmodulin‐de‐pendent protein phosphatase calcineurin (CaN). The inhibition of CaN blocks the activation of genes of IL‐2, IL‐2R, IL‐4, etc. in T cells. In addition, immunosuppressive and/or antiinflammatory activity of CsA can be executed via CyP‐40 and hsp70 complexed with SR, and following the interaction with CyP‐related receptor of NK and with IL‐8. CsA binding to CyPC, P‐GP and actin may throw light on the biochemical events leading to nephrotoxicity and graft vessel disease, two major side effects produced by CsA. The discovery of the interaction of human immunodeficiency virus type 1 (HIV‐1) Gag protein with CyP and effective disruption of this interaction by CsA may be important for our understanding of the pathology caused by this immunosuppressive virus and will inspire therapeutic strategies to nip HIV in the bud. Bacterial immunophilins (ImPs) contribute to the virulence of pathogenic microorganisms. Elucidation of molecular mechanisms of microbial ImPs' action in the pathogenesis of bacterial infections may lead to new strategies for designing antibacterial drugs.
Evaluation of Abbott TestPack RSV and an in‐house RSV ELISA for detection of respiratory syncytial virus in respiratory tract aspirates
Authors: Obel N. , Andersen H. K. , Jensen I. P. , Mordhorst C. H.
Published: August 2009

The Abbott TestPack RSV was compared with an in‐house RSV enzyme‐linked immunosorbent assay (ELISA) for detection of respiratory syncytial virus (RSV) antigen. Nasopharyngeal specimens were obtained from 121 inpatients. RSV antigen was detected in 46 specimens by the Abbott TestPack, 42 of these being confirmed by the in‐house RSV ELISA. Of the 75 specimens tested negative in the Abbott TestPack RSV, one was found positive by the in‐house RSV ELISA. The sensitivity and specificity of the Abbott TestPack RSV versus the RSV ELISA were 98% and 95% respectively.
Antibodies to Helicobacter pylori in dyspeptic patients, asymptomatic adults, and children from Costa Rica
Authors: Bartels GISELLE , Herrera ADELITA , Salas PILAR , Sierra RAFAELA , Lomonte BRUNO
Published: August 2009

An enzyme immunoassay for the detection of serum IgG antibodies to H. pylori whole bacterial sonicate was used in a study of dyspeptic adult patients (n=165), apparently healthy blood donors (n=154), and children (n = 66) from Costa Rica, where a very high incidence of gastric carcinoma is observed. The mean antibody levels and frequency distributions were similar in the dyspeptic patients and the blood donors. Meanwhile lower antibody levels to H. pylori were more frequently observed in children than in adults, suggesting an age‐dependent increase in seroprevalence, as described in studies from other populations.
Tumor necrosis factor α plus interleukin 1β treatment protects granulocytopenic mice fromPseudomonas aeruginosalung infection: Role of an unusual inflammatory response
Authors: Amura CLAUDIA R. , Fontan PATRICIA A. , Sanjuan NORBERTO , Nociari MARCELO M. , Buzzola FERNANDA R. , Sordelli DANIEL O.
Published: August 2009

We have recently demonstrated that treatment with interleukin 1β (IL‐1β) plus tumor necrosis factor a (TNFα) protects granulocytopenic hosts fromPseudomonas aeruginosaaerosol challenge. In this study we characterized the inflammatory response induced byP. aeruginosain granulocytopenic mice treated with 2,000 U IL‐1β plus 2,000 U TNFα. Treatment with the nonsteroidal anti‐inflammatory agent piroxicam abolished both the protective effect of cytokine treatment and the increase in myeloperoxidase (MPO) pulmonary activity. Histopathological studies revealed that, after aerosol challenge withP. aeruginosa, treatment with these cytokines induced migration and extravasation of mononuclear cells of immature appearance into the lung parenchyma. These cells contained MPO in their cytoplasm and displayed phagocytic capacity. Resident alveolar macrophages exhibited signs of activation and appeared in reduced numbers in bronchoalveolar lavage fluid. We suggest that the inflammatory response promoted by low TNFα plus IL‐1β doses may be one mechanism responsible for protection of granulocytopenic hosts fromP. aeruginosaaerosol challenge.
Evaluation of a method for measurement of intracellular killing of Staphylococcus aureus in human neutrophil granulocytes
Authors: Nielsen SØREN L. , Black FINN T. , Storgaard MERETE , Obel NIELS
Published: August 2009

A modified method for measurement of intracellular killing of Staphylococcus aureus in human neutrophil granulocytes is described. After phagocytosis of S. aureus the extracellular bacteria were sufficiently removed by repeated centrifugations and washings of the granulocytes. The optimal conditions for incubation of granulocytes for measurement of intracellular killing were found to be 37°C in the presence of 5% CO2. Under these conditions, stable pH, the viability and the capacity of the granulocytes for superoxide anion generation were preserved. The number of intracellular viable bacteria was determined after lysis of the granulocytes, which should be done in H2O at pH 11 to ensure sufficient cell lysis. The kinetics and individual variation of the intracellular killing are described. The intra‐ or extracellular location of surviving bacteria was studied. After approximately 8 h incubation we observed intracellular growth of S. aureus followed by lysis of granulocytes and extracellular growth of bacteria. Consequently, the incubation period should not be extended beyond 5 to 8 h when the assay is used to study the effects of antibiotics on intracellular killing.
The role of heparan sulphate‐binding activity of Helicobacter pylori bacteria in their adhesion to murine macrophages
Authors: Chmiela M. , Paziak‐Domanska B. , Rudnicka W. , WadstrÖM T.
Published: August 2009

The aim of this study was to determine the role of heparan sulphate (HS)‐binding activity of Helicobacter pylori microbes in their adhesion to and ingestion by inflammatory peritoneal macrophages. Two H. pylori strains expressing sialic acid‐specific haemagglutinins but differing in the expression of heparan sulphate‐binding capacity were chosen for investigation. The attachment to an ingestion by macrophages of the H. pylori bacteria were estimated by ELISA using anti‐H. pylori antibodies. The adhesion of both H. pylori strains could be inhibited by pretreatment of the bacteria with heparin (H), HS or fetuin, as well as by preincubation of the macrophages with heparinase or neuraminidase. However, detailed analysis of the data on the inhibition of bacterial adhesion to macrophages led to the conclusion that the attachment of H. pylori 25 bacteria, which expressed a high heparan sulphate binding, was mainly determined by HS‐binding structures. In contrast, the adhesion to macrophages of H. pylori bacteria 17874 microbes, which expressed a weak heparan sulphate binding, was more dependent on the exhibition of sialic acid‐dependent haemagglutinins. The described variation in H. pylori bacterial surface structures mediating their adhesion to macrophages could suggest a similar variation in bacterial adhesion to stomach mucosa and maybe in the pathogenicity of H. pylori strains.
Self‐induced bacteremia
Authors: Kristensen BRIAN , Tingsgaard LONE K. , Ejlertsen TOVE
Published: August 2009

We describe a patient with perforated appendicitis who postoperatively suffered repeated episodes of shaking chills and temperature spikes. Initial blood cultures yielded growth of Flavobacterium meningosepticum, Pseudomonas putida and Pseudomonas paucimobilis, and succeeding blood cultures growth of Pseudomonas acidoverans. These bacteria in combination led to a suspicion of self‐inoculation of contaminated water through an intravenous catheter. Antibiotic treatment had no effect on the symptoms, which only ceased when the intravenous catheter was removed.
Pathology of the thymus: Changes induced by xenobiotics and gene targeting
Authors: SCHUURMAN HENK‐JAN , KUPER C. FRIEKE
Published: August 2009

Studies on the thymus in pathologic conditions have been of great help in the elucidation of the function of the organ in T‐cell development. The first examples come from congenital immunodeficiency states in man and laboratory animals. A number of toxic substances affect different components of the thymus already at exposure levels where there is no effect on the peripheral immune system. In some cases, this thymotoxic effect has been causally related to defects in the peripheral immune system (immunodeficiency and autoimmunity). In recent years immunodeficient states have been created in mouse by disruption of genes coding immunologically relevant molecules. Studies on such gene ‘knock‐out’ mice have shown that a number of molecules are indispensable for appropriate T‐cell development at different stages in the thymus, whereas others are dispensable. It is concluded that the experimental approach combining gene targeting and exposure to thymotoxic xenobiotics will present interesting tools for further studies in thymus research.
Interobserver agreement for tumour type, grade of differentiation and stage in endometrial carcinomas
Authors: NEDERGAARD LOTTE , JACOBSEN MARIANNE , ANDERSEN JØRN ERIK
Published: August 2009

The histopathologic evaluation plays a major role in subdividing endometrial carcinomas into treatment groups. We have evaluated the interobserver agreement regarding tumour type, grade of differentiation, stage and stage I low and high risk cases. A total of 177 cases of endometrial carcinoma in which a hysterectomy and a bilateral salpingo‐oophorectomy were performed, were reviewed by three examiners. A variety of features including tumour type, architectural grade, nuclear grade, FIGO grade, and spread/metastases were recorded, and the FIGO stage was determined. Using two different definitions low and high risk groups in stage I tumours were separated. A kappa value was calculated for each of the various parameters. The current study showed a good strength of agreement for tumour type, myometrial invasion, spread/metastases, and FIGO stage (kappa 0.62–1.00). For two of the examiners good agreement was found as to architectural grade (kappa 0.71) while the kappa value for nuclear grade was lower (0.56). As nuclear grading is included in the revised FIGO recommendation a precise definition of nuclear atypia is needed. In stage I tumours very good agreement was demonstrated as to the defined low and high risk group (Kappa 0.64–0.86).
HLA antigens in ulcerative colitis and primary sclerosing cholangitis
Authors: LEIDENIUS MARJUT H. K. , KOSKIMIES SAIJA A. , KELLOKUMPU ILMO H. , HÖCKERSTEDT KRISTER A. V.
Published: August 2009

The aims of this study were to find out whether the alleles of the HLA class I or II region are associated with susceptibility to ulcerative colitis, and to show whether there is a difference or similarity in HLA associations between primary sclerosing cholangitis and ulcerative colitis. HLA‐A, B, C and DR antigens were studied using the standard lymphocyte microcytotoxicity test in 24 Finnish patients with primary sclerosing cholangitis, 77 patients with ulcerative colitis, and 106 controls. HLA‐B8 (54%) and DR3 (60%) were associated with primary sclerosing cholangitis. HLA‐DR1 (46%) and DR6 (20%) seemed more common in ulcerative colitis than in controls. A positive association with Cw7 was common to both ulcerative colitis (25%) and primary sclerosing cholangitis (33%). Our results indicate that ulcerative colitis is more heterogeneous than primary sclerosing cholangitis in its HLA‐DR associations.
Six billion neurons lost in AIDS
Authors: OSTER SOLVEIG , CHRISTOFFERSEN PER , GUNDERSEN HANS‐JØRGEN G. , NIELSEN JENS OLE , PEDERSEN COURT , PAKKENBERG RENTE
Published: August 2009

Human immunodeficiency virus type 1 (HIV1) is neurotropic. One of the morphological changes that is seen in patients with acquired immunodeficiency syndrome (AIDS) is cerebral atrophy affecting various structures including the neocortex. The cause of atrophy is not known. The total number of neocortical neurons was estimated in formalin fixed brains of 12 males with AIDS and 12 male controls matched for age and height. The mean number of neocortical neurons was 16.0times109 (coefficient of variation=0.11) in the AIDS patients compared with 21.9times109 (coefficient of variation=0.22) in the controls, a difference of approximately six billion (p<0.005, 2‐tailed). The global neuronal loss was 37%, and affected all four neocortical lobes. Ten patients did not have a history of central nervous system symptoms; two patients had a history of dementia. The number of neurons in the AIDS cases was not associated with dementia. AIDS is the first disease in which a global loss of neocortical neurons has been demonstrated using unbiased stereological methods. The loss of more than one third of the neurons may partly explain the cortical atrophy. Focal neuron loss has been reported by several authors, but none have been based on unbiased methods. In this group of AIDS patients the severe loss of neurons did not correspond to neurological deficits.
Development of a peroxidase‐antiperoxidase (PAP) technique for the identification ofHaemophilus somnusin pneumonic calf lungs in Denmark
Authors: TEGTMEIER CONNY , JENSEN NIELS E. , JENSEN HENRIK E.
Published: August 2009

A peroxidase‐antiperoxidase (PAP) technique was developed for the identification ofHaemophilus somnusbacteria in lung tissues of calves. Antisera raised against somatic and wall antigens of a Danish and American strain ofH. somnuswere produced. Experimentally infected murine tissues were used for the determination of the sensitivity and specificity of antiserum that had been heterologously absorbed with antigens of cross‐reacting bacteria, i.e.Pasteurella haemolyticaandPasteurella multocida.None of the antisera reacted withActinomyces pyogenes.An antiserum raised against somatic antigens of the Danish strain ofH. somnusrevealed the highest sensitivity in the PAP technique and became specific following absorption. Heterologous absorption also rendered this antiserum specific in crossed immunoelectrophoresis. Subsequently, the PAP technique was applied on formalin‐fixed pneumonic lung tissues of 86 calves. An immunodiagnosis ofH. somnuspneumonia was obtained in 15 of 17 lungs from which the bacterium had been isolated. Moreover, immunostained bacteria were also demonstrated in 20 lungs from whichH. somnushad not been isolated. Thus, application of immunohistochemistry significantly enhanced the diagnostic sensitivity ofH. somnuspneumonia of calves and should be used as a potent supplementary tool for the routine screening of suspected lung tissues of calves from which bacterial isolation is negative.
Sertoli cells of intratubular germ cell neoplasia express β2 microglobulin
Authors: BRÆNDSTRUP OTTO , MØLLER MAJ‐LIS , WERDELIN OLE
Published: August 2009

The cells in intratubular germ cell neoplasia in the vicinity of 38 germ cell tumors of the testis, including 20 pure seminomas, were studied for the expression of β2‐microglobulin (β2m), the constant component of all HLA class I molecules. Immunohistochemistry using antibodies towards β2m, vimentin, placental alkaline phosphatase, and ferritin was employed. Whereas the intratubular cells in normal testis are β2m negative, β2m positive cells were identified in intratubular germ cell neoplasia tubules in 55 per cent of all tumors and in 60 per cent of the seminomas. The tubules with β2m positive cells were located in areas with invasive tumor or in the vicinity of such areas. The β2m positive cells were identified as Sertoli cells by morphology and by their staining with anti‐vimentin. Neoplastic germ cells, identified by morphology and staining with anti‐placental alkaline phosphatase and anti‐ferritin were β2‐microglobulin negative. The most intensely β2m‐stained Sertoli cells were found in tubules with high concentrations of neoplastic germ cells. Intensely stained Sertoli cells were also found in ‘Sertoli cell only’ tubules inside invasive tumors and in areas without lymphocytic infiltration. The cells in adjacent normal tubules were β2m negative.
β2‐microglobulin expression of AIDS‐related and classical Kaposi's sarcoma
Authors: PETERSEN BODIL LAUB , PETERSEN CLAUS LETH , JENSEN ANNETTE , THOMSEN HENRIK KLEM , WERDELIN OLE , BRÆNDSTRUP OTTO
Published: August 2009

The expression of β2‐microglobulin, the invariable light chain of HLA class I molecules, of Kaposi's sarcoma from 11 AIDS patients and from 11 patients without known immunodeficiency was studied by immunohistochemistry using a polyclonal antibody to β2‐microglobulin. The staining intensity of spindle cells in these lesions was scored in a semiquantitative system. We found that the spindle cells of Kaposi's sarcomas from AIDS patients showed significantly increased staining intensity for β2‐microglobulin compared to those of Kaposi's sarcomas from non‐AIDS patients. The results may indicate that Kaposi's sarcomas developing in immunocompromised individuals, such as AIDS patients, are not subject to immune selection by T cells eliminating HLA class I high‐expressing tumor cells, while this may be the case in non‐AIDS patients. Alternatively, the results may be caused by differences in the activity of cytokines, which upregulate the expression of HLA class I molecules on the cell surface.
Metallothionein expression in placental tissue in Menkes' disease
Authors: HÆRSLEV TORBEN , JACOBSEN GRETE KRAG , HORN NINA , DAMSGAARD ELSE
Published: August 2009

Menkes' disease is a recessive X‐linked disturbance of copper metabolism, resulting in accumulation of copper in several extra‐hepatic tissues including the placenta. Metallothionein (MT) is a low‐molecular weight protein with a high affinity for group II metal ions, such as copper. Its synthesis is induced by the presence of the ions. The aim of this study was to investigate the pattern of the MT immunoreactivity in placental tissue obtained from women at‐risk of Menkes' disease in order to examine whether the MT occurrence and distribution may reflect the copper content. Placental tissue from six women with a family history of Menkes' disease, from 4 women without a family history, and from 2 hydatiform moles was studied. Positive MT immunostaining was found to be independent of the length of fixation, whether the tissue samples were fixed in 4% buffered formaldehyde or Bouin's fixative. The avidin‐biotin‐complex (ABC)‐technique was used. The copper content was measured by neutron activation analysis (NAA). In all placental tissue sections positive MT immunostaining appeared only in the trophoblast and only in proliferating cells. In placental tissue sections obtained from foetuses and children affected by Menkes' disease an additional MT immunostaining appeared in the Hofbauer cells of the chorionic villi. This staining was associated with an increased content of copper as measured by NAA. We conclude that the immunohistochemical demonstration of MT reflects the copper content and may be useful in pre‐ and postnatal diagnosis of Menkes' disease.
Postmortem axial skeletal radiography can reveal fetal CNS malformations
Authors: KJER INGER , HANSEN BIRGIT FISCHER
Published: August 2009

The routinely performed autopsy of a macerated fetus will often be of dubious value, particularly as regards the examination of the central nervous system (CNS). Former studies have demonstrated a close relationship between certain CNS malformations and axial skeletal malformations revealed radiographically. In the present report a postmortem examination of a severely macerated fetus demonstrates a transsphenoidal encephalocele. A supplementary histological examination confirmed this condition and furthermore revealed absence of the pituitary gland. The findings were related to a strongly elevated serum alphafetoprotein level in the 18th gestational week. This report emphasizes the value of postmortem axial skeletal radiography of autolyzed fetuses suspected for CNS malformations.
Morphometric studies on treated and untreated heterotransplanted malignant tumours
Authors: ALBRECTSEN J. , RYGAARD J.
Published: August 2009

Short‐term therapy effects on heterotransplanted tumours are important to know in order to study the action of anticancer treatment. They are, however, difficult to assess due to interference from such factors as edema, vascular swelling and necrosis. Non‐invasive methods such as NMR can be of some use, but the applicability is limited since background noise from the host animal organism may confound measurements. We studied a series of Cisplatine‐treated bladder carcinomas heterotransplanted to nude mice and their untreated controls in order to illustrate short‐term chemoterapy effect. We applied simple unbiased morphometric methods on formalin‐fixed and paraffin‐embedded material in order to obtain an unbiased estimate of the volume of the whole tumour and the volume of vital tumour tissue on day 1 and day 3 after treatment, at which time no gross changes in tumour size were evident. Furthermore, we estimated the number of tumour cells in the tumours. The estimate was based on measurement of the mean nuclear volume and the nuclear volume fraction in combination with the measured volume of vital tissue in the tumour. The pronounced therapy effects seen in this experimental system may be of potential interest in monitoring effects at the clinic.
Expression of the intercellular adhesion molecule‐3 (ICAM‐3) in human renal tissue with relation to kidney transplants and various inflammatory diseases
Authors: KNUDSEN H. , ANDERSEN C. B. , LADEFOGED S. D.
Published: August 2009

Adhesion molecules are important for immune regulatory mechanisms concerning antigen presentation, lymphocyte activation, localisation and migration as well as effector‐target cell interactions in inflammatory processes. The immunohistochemical expression of ICAM‐3, a recently cloned new member of the immunoglobulin family which also binds leucocyte function antigen 1 (LFA‐1), was examined in 80 needle core biopsies from 35 renal allografts, 7 patients with mesangioproliferative glomerulonephritis, 5 patients with extracapillary glomerulonephritis, 4 patients with interstitial nephritis and 5 patients with diabetic nephropathy and 20 normal kidneys. In all types of lesions ICAM‐3 was constitutively expressed on the majority of infiltrating leucocytes without detectable upregulation or presentation on possible target structures during inflammation indicating its possible role to be mainly in initiation of the inflammatory response.
Primary paraganglioma of the lung Report of two cases and review of the literature
Authors: SKØDT VERA , JACOBSEN GRETE KRAG , HELSTED MORTEN
Published: August 2009

Two cases of primary paragangliomas of the lung are presented. The first occurred in a 69‐year‐old woman and was a grossly and histologically benign tumour. The other case occurred in a 33‐year‐old woman who had metastases to the peribronchial lymph nodes. This seems to be the second malignant case reported. No evidence of recurrence nor metastatic disease was found at follow‐up, 8 months for the first case and 7 years for the second, respectively. The morphological and immunohistochemical findings are discussed and the literature comprising 20 cases is reviewed.
A case of cerebral paragonimiasis in Denmark
Authors: MØLLER A. , SETTNES O. P. , JENSEN N. O. , KRUSE‐LARSEN C.
Published: August 2009

A case of cerebral paragonimiasis with severe neurological symptoms is presented. The patient, a 45‐year‐old woman, recovered completely after resection of a large cyst at the C3 level. The pathogenesis is discussed.
Fastidious Gram‐negative bacteria: Meeting the diagnostic challenge with nucleic acid analysis
Authors: TØNJUM TONE , BØVRE KJELL , JUNI ELLIOT
Published: August 2009

The extended panorama of fastidious Gram‐negative bacteria (FGNB) as opportunistic etiological agents of infectious diseases in immunocompromised patients is largely due to improved medical expertise and technology. The heightened awareness of infectious diseases due to FGNB species mandates comprehensive classification and identification systems as a basis for rapid and reliable diagnostics. The most useful approaches are combinations of nucleic acid techniques such as hybridization, genetic transformation, amplification and base sequence analysis with selected conventional criteria. Among these approaches, the widely distributed feature of natural competence in these organisms facilitates the use of the biological method of genetic transformation as a valuable addition to the more common nucleic acid techniques. We describe the development of the taxonomy of FGNB through the last four decades, with particular emphasis on the families Neisseriaceae, Moraxellaceae, and Pasteurellaceae.
An electron microscopic study of clinical Paget's disease of the nipple
Authors: JAHN HENRIK , OSTHER PALLE JÖRN , NIELSEN ELLEN HOLM , RASMUSSEN GURLI , ANDERSEN JOHAN
Published: August 2009

An ultrastructural study of the epidermis from eight patients with clinical Paget's disease of the nipple supports the epidermotropic theory. There was no evidence that the Paget's cells originated from keratinocytes. We propose the hypothesis that Paget's cells represent transformed ductal cells, from the subjacent lactiferous ducts of the nipple, which have migrated into the epidermis, either as neoplastic cells or as normal ductal cells with secondary in situ transformation in the epidermis.
Intermediate filament expression and lectin histochemical features of canine transmissible venereal tumour
Authors: GIMENO E. J. , MASSONE A. R. , MARINO F. P. , IDIART J. R.
Published: August 2009

Immunocharacterization of intermediate filament proteins and lectin‐binding studies were carried out in canine transmissible venereal tumour (TVT), a unique neoplasia sharing some epidemiological features with Kaposi's sarcoma in humans. Neoplastic cells express vimentin, but neither cytokeratin nor desmin. Regarding lectins, TVT cells express receptors for Triticum vulgaris (WGA), Concanavalia ensiformis (Con A) and Ricinus communis I (RCA‐I). They appear to be negative for Ulex europaeus‐I (UEA‐I), Arachis hypogaea (PNA), Glycine maximus (SBA) and Dolichos biflorus (DBA).
Bcl‐2 oncoprotein is widespread in lymphoid tissue and lymphomas but its differential expression in benign versus malignant follicles and monocytoid B‐cell proliferations is of diagnostic value
Authors: WANG TSAILING , LASOTA JERZY , HANAU CHERYL A. , MIETTINEN MARKKU
Published: August 2009

The distribution of Bcl‐2 oncoprotein was studied immunohistochemically in formaldehyde‐fixed and paraffin‐embedded reactive and neoplastic lymphoid tissue. The potential of Bcl‐2 for the differential diagnosis of follicular lesions was emphasized, and the results on follicular lesions were correlated with those of polymerase chain reaction (PCR) assay of the immunoglobulin heavy chain gene rearrangement. In hyperplastic lymphoid tissue, Bcl‐2 reactivity was widespread, including germinal center surroundings, scattered cells within the germinal centers, and the T‐cell areas in general. Distinctively negative lymphoid populations included the majority of germinal center cells, and the negative staining pattern was maintained in cases of florid hyperplasia. In contrast, follicular lymphoma cells were consistently Bcl‐2 positive. The immunohistochemical Bcl‐2 reactivity of lymphoma follicles correlated with the clonal PCR amplification pattern of the immunoglobulin heavy chain gene; all Bcl‐2‐negative hyperplasias revealed a non‐clonal pattern. Clusters of monocytoid B cells were Bcl‐2 negative, whereas monocytoid B‐cell lymphomas and closely related MALT lymphomas were positive. All other small cell non‐Hodgkin's lymphomas of B‐cell types showed nearly uniform Bcl‐2 reactivity, whereas large cell B‐cell lymphomas were variably positive (74%). In Hodgkin's cells, Bcl‐2 reactivity was seen in the neoplastic populations of most cases of nodular sclerosis and mixed cellularity types, whereas the L&H and Reed‐Sternberg cells in lymphocyte predominance Hodgkin's disease were negative in most cases. Bcl‐2 immunohistochemistry thus appears very valuable in the differential diagnosis of follicular hyperplasia and neoplasia, and it may help to distinguish between reactive and neoplastic monocytoid B cells. However, Bcl‐2 immunohistochemistry is not useful in the subtyping of B‐cell lymphomas.
Typing of coagulase‐negative staphylococci from peritonitis in CAPD‐patients by the PhP‐CS system and REA
Authors: JUNG K. , BRAUNER A. , KÜUHN I. , RANSJÖO U. , HYLANDER B. , FLOCK J.‐I. , MÖOLLBY R.
Published: August 2009

Coagulase‐negative staphylococci (CNS) were the most common bacteria causing peritonitis in patients treated with continuous ambulatory peritoneal dialysis (CAPD). In order to investigate if the same clone was responsible for the peritonitis in the different patients and if the exit site was the source of infection we followed 68 patients on CAPD for 2 years. During this period 9 patients had 12 episodes of peritonitis caused by CNS. Cultures were taken from exit site and peritoneal fluid in all patients at peritonitis and during the first study year at monthly intervals. In each culture up to 10 isolates of CNS were randomly collected and frozen. All 437 CNS isolates from the patients with CNS peritonitis were typed using a biochemical typing method and 41 isolates identical by this method were further discriminated by a DNA fingerprinting method. Identical strains were in no case isolated from different patients, indicating that no virulent strain was spread between the patients. The isolates causing the peritonitis were never found at the exit sites before the first day of the peritonitis in any patient. In only two patients was the same strain found at the exit site and in the peritoneal fluid on the first day of peritonitis. It thus seems that no virulent clone of CNS was infecting the patients and we found no evidence of CNS at the exit site causing the peritonitis.
Survey of extraintestinal nontyphoid Salmonella infections in a Danish region
Authors: SCHØONHEYDER H. C. , EJLERTSEN T.
Published: August 2009

A total of 790 cases of nontyphoid salmonellosis distributed among 44 serotypes was registered in the County of Northern Jutland, Denmark (population 487,000) during a 3‐year period from January 1992 to December 1994. There were 45 cases of extraintestinal infection, 30 of which had a positive blood culture, 12 a positive urine culture, and 5 a positive culture of either pus or tissue. For the 9 serotypes represented by ≥5 isolates the proportion of extraintestinal isolates was inversely related to overall frequency of isolation (chi‐square test, f=1, p=0.00003). Although limited, our study is consistent with surveys in the UK and US pointing to a modifying effect of exposure on the invasiveness of nontyphoid Salmonella serotypes.
Serodiagnosis of Helicobacter pylori infection in patients with human immunodeficiency virus infection
Authors: NIELSEN HENRIK , ANDERSEN LEIF P.
Published: August 2009

In contrast to the established role of Helicobacter pylori gastritis in gastritis and duodenal ulcer in general, conflicting results have been reported in patients with human immunodeficiency virus (HIV) infection and the acquired immunodeficiency syndrome. The seroprevalence during early HIV infection is unknown. We retrospectively studied 102 patients with HIV infection early during the infection and in most cases in asymptomatic patients. Serological IgG antibody response to H. pylori was assessed by ELISA. Compared with an age‐matched control group the seroprevalence of H. pylori positivity was not significantly different (19% vs 25%). We observed no association with CD4 counts, p24 antigen, antibiotic prophylaxis with sulfamethoxazole/trimethoprim or antiretroviral treatment. In 54 of 83 patients initially seronegative a second examination was performed after a median of 24 months (range 3–60 months) and 2 patients had H. pylori seroconverted, indicating an incidence of new infection of 2%/year. In conclusion, previous reports have underestimated the prevalence of H. pylori infection in HIV patients, which seems to be similar to that in an HIV‐negative population.
Intracellular and surface distribution of CD9 in human eosinophils
Authors: FERNVIK EVA , HALLDÉEN GUNILLA , HED JAN , LUNDAHL JOACHIM
Published: August 2009

Expression of CD9 is a feature of both eosinophils and platelets. We have investigated the CD9 expression on resting and activated eosinophils with regard to possibly interacting platelets. Mixed leukocytes were obtained from the platelet‐containing (PC) and platelet‐depleted (PD) peripheral blood of healthy donors. A cell membrane permeabilization technique, the FOG method, enabled us to detect the eosinophils as a separate population and permitted flow cytometric analysis of both surface and intracellular antigens. Monoclonal antibodies against CD61 were used to identify platelets. The CD9/CD61 ratio indicated that CD9 on resting eosinophils originates mainly from eosinophils and not from adhered platelets. No difference in CD9 expression was obtained between resting eosinophils in PC and PD blood. However, the expression of CD9 was decreased (p<0.05) on eosinophils in PMA‐activated PD blood but increased (p=0.001) in PC blood, probably due to interacting platelets since CD61 increased simultaneously. In addition, we were able to detect an intracellularly stored pool of CD9 in eosinophils which decreased after activation with PMA. Together these results indicate a translocation of intracellularly stored CD9 to the cell membrane upon activation, probably followed by a subsequent shedding.