bluuebunny/crossref_metadata_2025 huggingface dataset:
Improved identification of Staphylococcus aureus using a new agglutination test Results of an international study
Authors: CROIZE J. , GIALANELLA P. , MONNET D. , OKADA J. , ORSI A. , VOSS A. , MERLIN S.
Published: August 2009

A new reagent for the identification of Staphylococcus aureus, SLIDEX STAPH‐KIT, operates on the principle of a latex and red blood cell combination agglutination: red blood cells are coated with fibrinogen for the detection of clumping factor, and latex particles are sensitized with anti‐S. aureus serotype 18 monoclonal antibody for the detection of protein A and antigen 18. French strains belonging to serotype 18 are methicillin‐resistant. The performance of this reagent was compared with STAPHYSLIDE and STAPHAUREX in Europe (France, Germany, Italy), in the United States and in Japan using 548 methicillin‐resistant S. aureus strains, 392 methicillin‐sensitive S. aureus strains, and 441 non‐aureus staphylococci. The specificity of the three reagents was equivalent (98.8% for SLIDEX STAPH‐KIT, 99,1% for STAPHYSLIDE, 98.1% for STAPHAUREX). SLIDEX STAPH‐KIT (97.3%) was more sensitive than STAPHYSLIDE (93.5%) and STAPHAUREX (89.7%) for all S. aureus strains due to a higher rate of identified methicillin‐resistant S. aureus strains.
Superantigens
Authors: Fleischer BERNHARD
Published: August 2009

“Superantigens” is the term for a group of molecules that have in common an extremely potent stimulatory activity for T lymphocytes of several species. They stimulate CD4+, CD8+ and gammaφ+ T cells by a unique mechanism: they cross‐link variable parts of the T‐cell receptor (TCR) with MHC class II molecules on accessory or target cells. The interaction site on the class II molecule and on the TCR is different from the peptide binding site; on the TCR it is the variable part of the ß chain (Vß). The prototype superantigen is the staphylococcal enterotoxin B (SEB), member of a family of genetically related proteins produced by Staphylococcus aureus and Streptococcus pyogenes. These are soluble exotoxins of approximately 27 kd molecular mass. It is intriguing that this molecular mechanism of T‐cell stimulation has been independently produced at least three times in evolution. Other pathogens producing superantigens are retroviruses (the Mouse Mammary Tumor Viruses) and a mycoplasma (Mycoplasma arthritidis). Many additional candidate superantigens have been proposed, but in most cases unequivocal evidence for superantigen activity is still missing. There are several reasons why these molecules have aroused such tremendous interest in recent years. First, they have provided key information on tolerance mechanisms, both on the deletion of T cells in the thymus and on the induction of peripheral tolerance by anergy and apoptosis. Second, of all polyclonal T‐cell stimulators they are the ones that most closely mimic the recognition of specific antigen. Finally, they have been recognized as important factors in the pathogenicity of the producing pathogens, inducing shock and immunosuppression. Whilst there is evidence that superantigens could be involved in the pathogenesis of certain human diseases, in most cases this is still very preliminary and indirect.
Human interferon‐γ quantified via a sensitive one‐site monoclonal antibody in a sandwich ELISA
Authors: Berg KURT
Published: August 2009

A new, specific and sensitive one‐site ELISA for precise quantification of human interferon‐gamma (HuIFN‐γ) at low levels in 50% human serum samples has been developed. The assay is based on the assumption that biologically active HuIFN‐γ is present exclusively as a dimer. Thus, in contrast to previous reports, the ELISA is based on a single monoclonal antibody (MAb) which is used in two ways: as “catching” antibody and as HRPO‐labelled conjugate. The sensitivity could be improved five‐fold by addition of (NH4)2SO4 to the conjugate solution; the lowest detectable amount of HuIFN‐γ is <0.5 u/ml. Non‐specific interactions were not seen in interferon samples taken from cultures, or samples diluted in ordinary media or 1% BSA. However, >30% of the (serum) samples gave non‐specific false‐positive results when the method was applied to samples containing 50–100% human serum from different donors. The false signals were related to the donors but could ‐ at the expense of the sensitivity which was reduced to 1 u/ml ‐ be abolished by PEG treatment of the (donor) serum samples.
Human papillomavirus and Epstein‐Barr virus in the etiology of testicular germ cell tumours
Authors: Meyts EWA RAJPERT‐DE , HØRding ULLA , Nielsen HENRIK W. , SkakkebæK NIELS E.
Published: August 2009

Epidemiological features suggest that the risk of testicular cancer may be related to exposure to unknown infectious agents, including viruses. Therefore a series of twenty specimens of testicular germ cell tumours, including preinvasive carcinoma in‐situ, were tested for the presence of DNA sequences of two viruses with known transforming abilities, human papillomavirus (HPV) and Epstein‐Barr virus (EBV). The polymerase chain reaction (PCR) technique was used. In none of the 19 successfully amplified samples were DNA sequences of HPV type 16 or type 18 detected. In six cases a faint trace of EBV DNA was revealed in one of two experiments. These samples were examined by imrnunohistochemical staining with specific antibodies raised against the EBV protein products and in‐situ hybridization with specific molecular probes, and were confirmed to be negative. The study indicates that a significant direct involvement of HPV and EBV in human testicular germ cell carcinogenesis is unlikely. However, a putative growth‐stimulating role of EBV‐transformed lymphocytes, which are frequently present in the stromal tissues of testicular tumours, cannot be excluded.
Novel automated microbial screening of platelet concentrates
Authors: Gong J. , HÖGman C. F. , Lundholm M. , Gustafsson I.
Published: August 2009

The recovery of bacteria and the speed of detection of microbial growth in platelet concentrates (PC) were studied, comparing a novel automated blood culture system (BacT/Alert) with two traditional methods. Full‐scale experiments were performed with a total of 33 units of PC (average content 320 × 109 platelets per unit) obtained from pooled buffy coats. Six strains known as possible PC contaminants were tested: S. epidermidis, S. aureus, P. aeruginosa, B. cereus, E. aerogenes, and S. sanguis. Using an inoculum of 50–98 bacteria/ml, bacterial contamination was detected in all of 11 PCs on 132 sampling occasions. With a small inoculum (0.3–9 bacteria/ml), the bacterial contamination was detected in all PCs in which abundant growth appeared, but failed to be detected in some instances where the bacterial content was very small. All of 11 uninoculated PC controls were negative. With the automated method the time for detection of a culture‐positive PC was 6–16 h, clearly shorter than with the two manual‐visual systems. The new system provides a rapid, reliable, and labour‐saving method for screening of bacterial contamination in PCs. This would increase safety particularly if the present 5‐day shelf life were extended.
Dichotomy in the human CD4+ T‐cell response to Leishmania parasites
Authors: Kemp MICHAEL , Kurtzhals JØRGEN A. L. , Kharazmi ARSALAN , Theander THOR G.
Published: August 2009

Leishmania parasites cause human diseases ranging from self‐healing cutaneous ulcers to fatal systemic infections. In addition, many individuals become infected without developing disease. In mice the two subsets of CD4+ T cells, Thl and Th2, have different effects on the outcome of experimental Leishmania infections. Thl cells producing interferon‐y (IFN‐y) mediate resistance, whereas Th2 cells producing interleukin‐4 (IL‐4) and IL‐10 are associated with susceptibility and exacerbation. Evidence is accumulating that a Th1/Th2 dichotomy in the T‐cell response to Leishmania exists also in humans, and that the balance between subsets of parasite‐specific T cells may play an important regulatory role in determining the outcome of the infections.
Distribution of cathepsin D in human polymorphonuclear and mononuclear blood cells
Authors: Barabasi K. , NÄSsberger L.
Published: August 2009

We have investigated the distribution of cathepsin D in mononuclear and polymorphonuclear cells using an anti‐rabbit human cathepsin D antibody and indirect immunofluorescence. Lymphocytes showed the same fluorescence intensity as control cells incubated with normal serum. Fluorescence staining patterns of polymorphonuclear cells differed according to the fixative used. Monocytes and granulocytes fixed in ethanol, methanol, glutaraldehyde, and Bouin's fixative caused a homogeneous cytoplasmic staining with a distinct nuclear sparing. A perinuclear staining was seen in acetone‐fixed cells in both monocytes and granulocytes. Another interesting difference was observed between monocytes and granulocytes when formalin was used as fixative. Monocytes appeared with a cytoplasmic staining, but in formalin‐fixed granulocytes redistribution to the nuclear membrane was seen.
Staphylococcus aureus fibronectin‐binding proteins (FnBPs)
Authors: Rozalska B. , Sakata N. , WadstrØM T.
Published: August 2009

Polyclonal antibodies against recombinant fibronectin‐binding proteins (gal‐FnBP A and ZZ‐FnBP B) of Staphylococcus aureus were analyzed by both solid‐phase and solution‐phase methods. These antibodies were found to bind homologous antigen and to cross‐react with heterologous antigen. It was also found that antibodies recognize native FnBP on the cell surface. It has been shown, by the inhibition assay, that the majority of antibodies recognize a fibronectin‐binding D1‐D2 sequence of FnBP A. Anti‐FnBP A Fab fails to bind the D3 sequence, though this peptide used in a solution inhibits binding of fibronectin to immobilized FnBP A, similarly to Dl and D2 peptides. Since the anti‐FnBP A antibodies are able to block fibronectin binding to staphylococci by about 50%, it is reasonable to assume that the bacterial receptor has additional binding sites outside the D domain.
A rapid density gradient technique for separating polymorphonuclear granulocytes
Authors: Cabanis ALICE , Gressier BERNARD , Lebegue SOPHIE , Brunet CLAUDE , Dine THIERRY , Luyckx MICHEL , Cazin MICHELINE , Cazin JEAN CLAUDE
Published: August 2009

A gradient separation technique followed by isotonic ammonium chloride haemolysis was compared with two methods for the isolation of polymorphonuclear neutrophils from blood. This technique provided a high yield, excellent purity without lymphocyte and erythrocyte contamination, and made it possible to isolate more than 50 times 106 human neutrophils from 15 ml of blood. The polymorphonuclear neutrophils isolated in this way were capable of generating a large amount of reactive oxygen species. This technique for the separation of polymorphonuclear neutrophils is an effective method for in vitro studies.
Human papillomavirus infection in Norwegian women with cervical cancer
Authors: Kristiansen EWY , Jenkins ANDREW , Kristensen GUNNAR , Ask EIRIK , KæRn JANNE , Abeler VERA , Lindqvist BJØRN H. , TropÉ CLAES , Kristiansen BJØRN‐ERIK
Published: August 2009

The objective of the present study was to determine the prevalence of human papillomavirus (HPV) infections in Norwegian women with cervical cancer. We used the polymerase chain reaction (PCR) and Southern blot techniques to assess the prevalence of HPV in cervical biopsies of 133 women admitted to the Norwegian Radium Hospital for treatment of cervical cancer. At the time of sampling (from February 1988 to April 1989) about 85% of Norwegian women with cervical cancer were treated at the Norwegian Radium Hospital. HPV was found in biopsies of 91 (68%) of women with cancer; 70 (53%) biopsies contained HPV type 16, 19 (14%) HPV type 18, 4 (3%) HPV type 33, 2 (1.5%) HPV type 11, and 3 (2%) HPV DNA of unknown type (HPVX). Five percent of biopsies were doubly infected, chiefly with HPV 16 + 18. We found a significant association between HPV 18 and low age, poorly differentiated tumors and adenocarcinomas. Our results show that there is an association between HPV types 16 and 18 and cervical cancer also in a Norwegian setting. PCR was more sensitive than Southern blotting for detection of HPV. Thirty‐six (27.5%) of cancer biopsies were positive by PCR but negative by Southern blotting, as against 49 (73.5%) positive by both methods; we also encountered 4 samples positive by Southern blotting and negative by PCR. In 23/53 cancer biopsies positive by Southern blotting we found evidence for integrated or rearranged HPV genomes.
Pathology of renal arteries of dyslipidemic children with congenital nephrosis
Authors: Antikainen M. , Sariola H. , Rapola J. , Taskinen M. R. , HÖLth6Fer H. , Holmberg C.
Published: August 2009

Congenital nephrotic syndrome of the Finnish type (CNF) is well characterized in infants and associated with major lipid risk factors for atherosclerosis. This study was undertaken to investigate if any arterial pathology is present in children with CNF and, if so, to describe its nature in renal arteries collected at nephrectomy at a mean age of 12.5 ± 4.4 months. Denuded endothelial injury and intimal thickening were seen in 9 out of 10 patient specimens of renal arteries. Intimal thickening contained loose abundant extracellular matrix with a few smooth muscle or myofibroblastoid cells. Only a few Sudan black‐ or oil red O‐positive lipid droplets were found in six and seven samples, respectively. Areas immunoreactive with antibodies against apoprotein B were seen in only two specimens. Immunohistochemistry did not reveal any activated T or B cells, or any expression of IL‐1 or IL‐2 receptors. Macrophages were present in only two specimens. No foam cells were seen. We conclude that the vascular pathology together with altered lipoprotein metabolism indicates that children with CNF might be at risk for early atherosclerotic arterial disease, particularly if their hyperlipidemia persists.
Amylase in lung carcinomas
Authors: Grove ANNI
Published: August 2009

The ultrastructure of two lung adenocarcinomas with immunohistochemical and biochemical expression of amylase is reported. Isoenzyme determination performed on tumour tissue showed a salivary‐type amylase. None of the carcinomas was associated with hyperamylasaemia. Ultrastruc‐turally, the adenocarcinomas contained a varying number of heterogeneous granules resembling lysosomes. None of the tumours contained mature zymogen granules. The findings are discussed in relation to previous cases of amylase‐containing lung carcinomas subjected to electron microscopy. The vast majority of these carcinomas were associated with a significant elevation of serum amylase. Various theories which might explain the absence of hyperamylasaemia in the presented cases are proposed. The biological and diagnostic significance of amylase production is discussed.
The modulation of tumour necrosis factor‐α, interleukin‐1α and glucose levels with GMDP and other analogues of muramyl dipeptide
Authors: Adeleye T. A. , Moreno C. , Ivanyi J. , Aston R.
Published: August 2009

Immunomodulatory agent muramyl dipeptide (MDP) and seven of its analogues were tested for ability to counteract the toxic actions of lipopolysaccharide (LPS) in experimental mouse models. Female BALB/c mice were presensitized with Corynebacterium parvum (P. acnes) and given MDP or equimolar doses of one of its analogues after 2 weeks, followed by intravenous challenge with LPS 18 h later. This treatment produced a sharp increase in blood cytokine (TNF‐oc, IL‐la) levels 4 h after LPS administration followed by a decline to control values after 6 h. Four analogues, GMDP, threonylMDP, GMDPBenz and GMDPOBut, were able to reduce the level of cytokines induced with LPS. For most of the analogues, the higher doses reduced the levels of TNF‐a but slightly increased the concomitant IL‐lα levels. GMDP was the most effective compound tested in terms of reduction of TNF‐α and IL‐lα levels, as well as for reduction of the hypoglycaemia caused by the administration of LPS.
Metachromatic, IgE‐bearing and tryptase‐containing cells on the nasal mucosal surface in provoked allergic rhinitis
Authors: Juliusson S. , Karlsson G. , Bachert C. , Enerback L.
Published: August 2009

We studied the relationship between mast cells or basophils and symptoms in provoked allergic rhinitis. Nasal brush and lavage samples were obtained before nasal allergen challenge and every 2 h for 12 h after the challenge in 10 allergies and 3 controls. The cells were identified by their metachromatic staining properties (brush and lavage samples) or with immunohistochemical methods using specific antibodies to IgE and tryptase, a selective mast‐cell marker (brush samples). Histamine was determined in the brush samples using liquid chromatography. After an initial decrease, the numbers of metachromatic, IgE‐bearing and tryptase‐containing cells, as well as the histamine content of the cells in the brush samples, increased during the subsequent observation hours. The prechallenge cell and histamine values correlated with the symptom score 15 min after the challenge. The prechallenge lavage samples lacked metachromatic cells, but these cells were found in increasing numbers after the provocation. Three of the patients differed from the remaining seven in that their prechallenge brush samples contained many positively stained cells. All patients showed a positive cellular response to the allergen challenge, but these three individuals showed the most vivid response. The morphology of the metachromatic cells in the prechallenge brush samples agreed with that of mast cells, but the morphology of metachromatic cells which outnumbered tryptase‐containing cells in samples at 8 to 12 h rather agreed with their being basophils.
An immunological study of chloralkali workers previously exposed to mercury vapour
Authors: Ellingsen DAG G. , Gaarder PER I. , Kjuus HELGE
Published: August 2009

Several animal studies have suggested that exposure to inorganic mercury compounds may have effects on the immune system. Human data are, however, relatively sparse. The presence of several autoantibodies and the concentration of immunoglobulins and complement components were determined in the present cross‐sectional study in 77 chloralkali workers previously exposed to mercury vapour and 53 age‐matched referents. The mean duration of exposure to mercury vapour was 7.9 years (range 1.1–36.2), and the exposure had ceased on average 12.3 years (range 1.0–35.0) prior to the examination. No increased prevalence of the studied autoantibodies was observed among the exposed subjects as compared with the referents. There were no significant differences in the concentrations of immunoglobulins or complement components between the two studied groups. Nor was any relationship observed between cumulative mercury dose estimates or time since cessation of exposure and the measured concentrations of immunoglobulins or complement components.
Atherosclerosis in Watanabe heritable hyperlipidaemic rabbits
Authors: Hansen BIRGIT FISCHER , Mortensen ALICJA , Hansen JØRGEN FISCHER , Ibsen PER , Frandsen HENRIK , Nordestgaard BØRGE G.
Published: August 2009

The spontaneous development of atherosclerotic disease in 38 homozygous and 34 heterozygous Watanabe heritable hyperlipidaemic rabbits was evaluated by qualitative and quantitative light microscopy in aorta, coronary, pulmonary and renal arteries, by naked eye and macroscopic morphometric estimation of aortic atherosclerosis extent and by biochemical analysis of aortic cholesterol content. No noteworthy atherosclerosis was demonstrated within 19 months in heterozygous rabbits. In homozygous rabbits, atherosclerotic lesions were seen from the age of 4 months and progressed with age. All 19‐month‐old rabbits had severe atherosclerotic disease. As much as 64% of the variation in atherosclerosis extent/severity could be explained by serum cholesterol and age. A highly significant correlation between the various methods for quantitation of atherosclerosis extent and/or severity was demonstrated, suggesting that quantitative microscopy, macroscopic morphometry and determination of aortic cholesterol content may be equally valid as a measure of atherosclerosis in WHHL rabbits and are therefore interchangeable.
An ultrastructural analysis of concordant and discordant cardiac xenografts in unmodified recipients
Authors: Nielsen BJARNE , SteinbrÜChel DANIEL A. , Kemp EJVIND
Published: August 2009

Ultrastructural changes in endothelial cells in a hamster‐to‐rat and guinea pig‐to‐rat heart transplantation model are described. In the hamster‐to‐rat model, changes were minimal with an increase in cytoplasmic vesicles 4–6 h after transplantation. 16–24 h after transplantation larger vesicles appeared and the basement and cell membranes were less well defined. 42–48 h after transplantation the changes had progressed with destruction of cell membranes and the appearance of extravasated erythrocytes. At the time of complete rejection, changes had further progressed with widespread endothelial cell destruction and infiltration of neutrophilic granulocytes and macrophages, and large amounts of fibrin were present. In the guinea pig‐to‐rat model, changes were characterized by the appearance of platelets in close contact with the endothelium of the capillaries 1–3 min after transplantation. 4–6 min after transplantation the basement membranes as well as the cell membranes were affected with indistinct borders and interruption. Occasionally fusion of platelets and endothelial cell membranes was demonstrated. In the grafts examined 7–9 min after transplantation, changes had further progressed. Massive aggregation of platelets now appeared in relation to remnants of endothelical cells. Signs of microvascular damage appeared in both models, but with different morphology. In hamster grafts, endothelial cell activation is indicated by gradual changes in the cell membranes resulting in vascular damage and infiltration of the grafts by macrophages and neutrophilic granulocytes. In the guinea pig grafts, activation of endothelial cells results in platelet aggregation, formation of microthrombi, and subsequent tissue damage. Even though antibody and complement are involved in both types of rejection the basic mechanisms are different.
The characterization of Danish isolates of Salmonella enterica serovar Enteritidis by phage typing and plasmid profiling: 1980–1990
Authors: Brown D. J. , Baggesen D. L. , Hansen H. B. , Hansen H. C. , Bisgaard M.
Published: August 2009

Plasmid profiling, phage typing and antimicrobial resistance typing have been carried out on 736 isolates of Salmonella enterica serovar Enteritidis collected in Denmark during the period 1980 to 1990. Strains originated from cases of human salmonellosis, broiler poultry flocks, layer poultry flocks, quarantined imported poultry, environmental samples from hatchery units, and from bovines. Phage type (PT) 1 was found to be the most common type among isolates of poultry origin (57.6%) followed by PT4 (28.8%). Isolates belonging to PT8 were found exclusively in imported birds. Phage typing of a representative sample of human isolates revealed the predominance, as in most other Western European countries, of PT4 (61.8%). PTI, however, was found in 17.0% of human strains, a much higher incidence than expected. Antibiotic resistance was observed in 4 out of 107 human isolates (3.7%) and 2 out of 205 non‐human isolates (1%) tested.
Restriction endonuclease analysis of adenovirus type 3 isolated in Norway from 1970 to 1991
Authors: Tveten YNGVAR , Ahlquist JøRN , Krogvold LARS , Bruu ANNE‐LISE , StØRvold GUNNAR , ØRstavik IVAR
Published: August 2009

Forty‐three strains of adenovirus type 3 isolated from patients in Norway between 1970 and 1991 were analyzed with four restriction endonucleases. Bgl II was the most discriminative enzyme. Five genotypes were identified and one of these has not been described before (Ad3a12). During both the epidemics in this period, new genotypes were introduced into the population. The same genotypes were identified in Norway as have previously been found in the northern parts of Europe, America and the Soviet Union.
Isoprinosine stimulates granulocyte chemiluminescence and inhibits monocyte chemiluminescence in vitro
Authors: FlØ REINHARDT W. , NæSs ARE , Albrektsen GRETHE , Solberg CLAUS OLA
Published: August 2009

Isoprinosine may delay disease progression in human immunodeficiency virus infection, presumably through modulation of lymphocyte function. However, the influence of isoprinosine on phagocyte function is largely unknown. This study describes the effects of isoprinosine and azidothymidine on phagocyte chemiluminescence and migration. Incubation with isoprinosine concentrations of 250 μg/ml and above increased the chemiluminescence of granulocytes. Random migration of granulocytes was decreased at isoprinosine concentrations of 50 μg/ml and higher, but chemotaxis was not affected. Azidothymidine exerted no effect on the chemiluminescence or migration of granulocytes. For monocytes, luminol‐enhanced chemiluminescence was reduced at isoprinosine concentrations of 250 μg/ml and above, whereas migration was not affected. These findings suggest that the immunomodulatory properties of isoprinosine may extend to phagocytic cells. This may be of significance in the treatment of immunodeficiency states.
Immunohistochemical reactivity in mesothelioma and adenocarcinoma: A stepwise logistic regression analysis
Authors: Dejmek ANNIKA , Hjerpe ANDERS
Published: August 2009

Histological sections from 103 malignant mesotheliomas and 43 adenocarcinoma metastases in pleural biopsies were investigated for reactivity against a panel of 11 different antibodies. The size of the material allowed the evaluation by stepwise logistic regression analysis, which selected five parameters of major importance: vimentin reactivity in epithelial cells, reactivity to low‐molecular‐weight keratins in fibrous cells, strong membrane accentuation of EM A reactivity, and lack of reactivity to LeuM1 and BerEp4. Three of these criteria were sufficient to identify a mesothelioma with high specificity and with a sensitivity of approximately 70%. Whilst the monoclonal anti‐CEA tested was the most valuable single parameter, it did not add any diagnostic information to the combination of criteria selected by the stepwise logistic regression analysis. However, this antibody can be used to exclude most of the adenocarcinomas from further analysis with the more extensive panel.
Bordetella pertussis diagnosed by polymerase chain reaction
Authors: BirkebæK N. H. , Heron I. , SkjØDt K.
Published: August 2009

The object of this work was to test the polymerase chain reaction (PCR) for demonstration of Bordetella pertussis (BP) in nasopharyngeal secretions. The method was applied to patients with recently diagnosed pertussis, as verified by BP culture. In order to test the sensitivity and specificity of PCR for the diagnosis of BP, we used known concentrations of BP, Bordetella parapertussis and Bordetella bronchiseptica in aqueous solutions. PCR was furthermore carried out on species of bacteria that might be isolated from the nasopharynx. The applicability of PCR to patient specimens was tested in 25 patients in whose nasopharyngeal secretions BP had been demonstrated after 4–7 days of culture. The detection limit of PCR in aqueous solution was 1–2 BP bacteria per reaction tube. PCR was 100% specific for BP, showing no response with other Bordetella species or other bacteria known to colonize the nasopharynx. Of 25 patient specimens, 16 were PCR‐positive 4–7 days after the positive primary culture had been established; only 5 out of 13 patient specimens were positive by repeated conventional nasopharyngeal culture at that time. We conclude that PCR is a possible alternative to culture for the demonstration of BP, as PCR is considerably faster than culture and might be more sensitive.
Alteration of immunoreactivity by hydrated autoclaving, microwave treatment, and simple heating of paraffin‐embedded tissue sections
Authors: Igarashi HISAKI , Sugimura HARUHIKO , Maruyama KEIJI , Kitayama YASUHIKO , Ohta ISAO , Suzuki MAKOTO , Tanaka MASAMITSU , Dobashi YOH , Kino ISAMU
Published: August 2009

The effects of treatment in a hydrated autoclave (121 °C, 2 atm for 20 min), microwave oven (in water), and simple heating (60 °C overnight in distilled water or 90 °C for 10 min in ZnSO4) on the stainability of 56 antigens by commercially available antibodies in formalin‐fixed paraffin‐embedded tissue sections were evaluated. The detectability of nuclear antigens, glycoprotein, lymphocytic surface markers, and chromogranin A was significantly and reproducibly improved by these treatments, whereas the detectability of viral antigens and peptide hormones was attenuated or unchanged. This enhancement includes not only the distinctiveness of the positive staining, but also the number of positive cells, as revealed by comparing serial sections. Among these four heating procedures, microwave heating and autoclaving were more effective than the others on p53, c‐erbB‐2, and CA125, whereas simple heating was best for smooth‐muscle actin (HHF35 and CGA7). Generally the effects of the heating procedures for these antigens were consistent among the cases, but the effects on GFAP varied with the case. The alterations we observed could significantly influence the interpretation of immunohistochemical staining of currently popular tumor markers such as p53 in terms of their prevalence (28%vs 64% in gastric cancer; 36%vs 82% in metastatic liver cancer) and other diagnostically important markers.
Priming of eosinophil and neutrophil migratory responses by interleukin 3 and interleukin 5
Authors: HÅKansson LENA , Venge PER
Published: August 2009

In the present study the influence of interleukin 3 and interleukin 5 on the migration of normal eosinophil and neutrophil granulocytes has been investigated. LTB4, PAF, f‐MLP, C5a and ZAS were used as chemoattractants, and HSA and pooled normal human serum were used as chemokinetic agents. Recombinant human IL‐5 (rh‐IL5) at a concentration of 4 times 10−12 mol/1 was chemotactic for eosinophils, while recombinant mouse IL‐5 (rm‐IL5) attracted both eosinophils and neutrophils. IL‐3 (rh‐IL3) at a concentration around 10−12 mol/1 exerted a priming effect on eosinophil and neutrophil migration, i.e. chemotactic and chemokinetic responses to all agents tested. Human IL‐5 at a concentration of 2 to 20 times 10−12 mol/1 primed the chemotactic and chemokinetic responses of eosinophils to all agents tested. The migration of neutrophils was also primed by rh‐IL5, but at higher concentrations, i.e. around 10−10 mol/1. IL‐5 of mouse origin primed the migration of both eosinophils and neutrophils. In conclusion, IL‐3 primed the migratory function of both eosinophils and neutrophils, while IL‐5 was a more potent primer of eosinophil than of neutrophil migration.
Treatment of shunt‐related cerebral ventriculitis due to Corynebacterium jeikeium with vancomycin administered intraventricularly
Authors: Knudsen JENNY DAHL , Nielsen CARSTEN JUUL , Espersen FRANK
Published: August 2009

A 52‐year‐old female with subarachnoid haemorrhage and hydrocephalus was treated with external and later internal drainage. She developed ventriculoperitoneal shunt‐related ventriculitis caused by Corynebacterium jeikeium. The infection was unsuccessfully treated with intravenous vancomycin. It was controlled only after shunt removal and administration of intraventricular vancomycin as well as systemic vancomycin, rifampicin and fusidic acid. A review of the literature confirmed our experience that vancomycin given intraventricularly is well tolerated and doses can be individualized by measuring vancomycin levels in cerebrospinal fluid.
Resistance characteristics of blood culture isolates of Enterobacter cloacae with special reference to beta‐lactamases and relation to preceding antimicrobial therapy
Authors: Weischer MERETE , Schumacher HELGA , Kolmos HANS JØRN
Published: August 2009

Resistance characteristics of 53 blood culture isolates of E. cloacae were examined and correlated with antimicrobial treatment preceding bacteraemia. Resistance patterns of 22 antimicrobial agents, presence of resistant mutants, and inducibility of beta‐lactamase were investigated; furthermore, population analysis and investigation of beta‐lactamase production of selected isolates were performed. Thirty‐two isolates (60%) were resistant to cephalothin and/or cefoxitin and/or ampicillin, and 14 isolates (26%) had further resistance characteristics, 7 of the 14 being resistant to non‐beta‐lactam antibiotics. All ampicillin‐susceptible and 76% of cefotaxime‐susceptible isolates had resistant mutants in the zone of inhibition when high inoculum was used. All isolates investigated had inducible chromosomal beta‐lactamases, and, in addition, two isolates had an enzyme corresponding with TEM‐1. Correlation of resistance patterns and antimicrobial treatment preceding bacteraemia showed that treatment with a third‐generation cephalosporin was associated with beta‐lactam multiresistance. In conclusion, susceptibility testing of beta‐lactam antibiotics of Enterobacter must be interpreted with caution and monotherapy with an extended‐spectrum cephalosporin should be avoided unless presence of resistant mutants and inducibility of beta‐lactamase can be excluded.
Epstein‐Barr virus in exfoliated cells from the postnasal space
Authors: HØRding ULLA , Albeck HENRIK , Katholm MORTEN , Kristensen HENNING SUND
Published: August 2009

There is a high incidence of undifferentiated nasopharyngeal carcinoma (NCP) in certain populations, including Greenland Eskimos. The cancer appears to have a causal association with Epstein‐Barr virus (EBV), which is regularly found in NPC epithelial cells. With the aim of developing a method of screening or facilitating early diagnosis of NPC, we used the polymerase chain reaction to examine exfoliated nasopharyngeal cells for EBV in 54 Greenland and 17 white Danish subjects, none of whom was suspected of having NPC. EBV DNA was found in 81% of Greenland and 35% of Danish subjects. These findings support the concept of EBV infection leading in most cases to a chronic carrier state. It is concluded that EBV detection in nasopharyngeal cells is not at present a suitable method for identification of individuals at increased risk of developing NPC.
Identification of coryneform and other Gram‐positive rods with several methods
Authors: VON Graevenitz ALEXANDER , PÜNter VERENA , Gruner EVA , Pfyffer GABY E. , Funke GUIDO
Published: August 2009

The identification of 202 isolates of aerobically growing Gram‐positive rods from clinical material was attempted by using a combination of “traditional” morphological and biochemical tests (Hollis & Weaver (20)) plus patterns of cellular and metabolic fatty acids. This system served as the “gold standard” for three others, i.e. API Coryne (Rapid Coryne), MIDI TSBA and MIDI CLIN Aerobic. In addition, several growth, biochemical and susceptibility tests (growth on cystine‐tellurite blood agar, DNase, hippurate and starch hydrolysis, methanethiol formation, API ZYM, CAMP reaction, susceptibility to O/129 and to six antimicrobials) were done in order to check their usefulness for the identification of this group of bacteria. Our system, with the help of chemotaxonomic tests (m‐DAP and mycolic acids), was able to identify 154/202 (76%) of the isolates by species and an additional 41/202 (21%) by genus only; 7 (3%) could not be identified. The API Coryne system identified to species or genus level 140/195 isolates (72%). Corresponding figures for the MIDI TSBA and CLIN systems were 63/195 (32%) and 88/195 (45%); further details of species and genus identification are presented in the text. The main drawback of the commercial systems is the extent and probably the numerical depth of the data base. We recommend the use of our multisystem approach for the identification of Gram‐positive rods until commercial systems are based on a broader and numerically more extensive data base. The additional tests did not prove species‐ or genus‐specific.
Thyroid antibodies in association with thyroid malignancy
Authors: Haapala A. M. , Soppi E. , MÖRsky P. , Salmi J. , Laine S. , Mattila J.
Published: August 2009

Occurrence of thyroid antibodies and thyroiditis in association with thyroid malignancy, suspected malignancy or other thyroid diseases was studied in 177 patients. Retrospective clinical analysis revealed that 137 patients had thyroid carcinoma (108 papillary carcinomas, 10 occult papillary carcinomas, 14 follicular carcinomas, and 5 other carcinomas) and 40 had other thyroid diseases. Thyroid microsomal (AMC) and thyroglobulin antibodies (ATG) were measured by the particle agglutination method. Clinically significant thyroid antibody titers and histological or clinical features of thyroiditis were seen in nine patients with thyroid carcinoma, being equally prevalent in follicular and papillary carcinomas. Taking into account the prevalence of positive AMC antibodies and thyroid carcinoma in Finland, the highest predicted prevalence of simultaneously occurring thyroid carcinoma and thyroiditis in the under 40 age group should be 0.002% and in the over 40 age group 0.006%. In the present material the observed occurrences in corresponding age groups were unexpectedly high (4.7% and 5.2%). The results may partly be due to patient selection, but they also suggest that there might be a link between thyroiditis and thyroid carcinoma.
Detection of the BC 24 transforming fragment of the herpes simplex virus type 2 (HSV‐2) DNA in cervical carcinoma tissue by polymerase chain reaction (PCR)
Authors: Yamakawa Y. , Forslund O. , Chua K. L. , Dillner L. , Boon M. E. , Hansson B. G.
Published: August 2009

The polymerase chain reaction (PCR) was used to investigate samples from Indonesian and Swedish patients with cervical intraepithelial neoplasia grade III (CIN III), squamous cell carcinoma or adenocarcinoma of the cervix for the presence of a transforming fragment (BC 24) of herpes simplex virus type 2 (HSV‐2) DNA. The PCR test for HSV‐2 DNA was more sensitive than the infectivity endpoint titer in a cell culture system and no cross reactivity was found with either varicella‐zoster virus, cytomegalovirus, Epstein‐Barr virus, human papillomavirus 16 or 18, or human genomic DNA. Using this PCR test, 2 out of 5 cases with CIN III, 10 of 71 squamous cell carcinomas, and 3 of 11 adenocarcinomas of the uterine cervix were found to contain DNA sequences homologous to the BC 24 fragment of the HSV‐2 genome. Only two of the samples containing this transforming region of the HSV‐2 DNA were positive in a PCR assay for the HSV‐2 DNA polymerase gene. The great majority of the HSV‐2 BC 24 DNA positive (12 of 15) came from the Indonesian group of patients. All 15 CIN III or cancer samples positive for the HSV‐2 BC 24 fragment were also positive for papillomavirus DNA. In line with observations made by others, our data support the hypothesis that HSV infection could represent one of several possible oncogenic cofactors leading to cervical carcinoma. The HSV cofactor might be more important in the Indonesian than in the Swedish population.
Calcium‐enhanced aggregation of serum amyloid P component and its inhibition by the ligands heparin and heparan sulphate
Authors: Nielsen E. HOLM , SØRensen I. JUUL , Vilsgaard K. , Andersen O. , Svehag S. E.
Published: August 2009

Serum amyloid P component (SAP) is a pentraxin found in the circulation and in all forms of amyloid deposits. Its physiological and pathophysiological functions are largely unknown. Electron microscopy showed purified human SAP to consist of double pentameric discs compatible with the results of size chromatography. The formation of double pentamers did not require calcium ions. The outer diameter of the discs arranged face‐to‐face was 11.6 nm and the inner diameter 3.2 nm. The thickness of single and double pentamers was 4.1 and 8.7 nm, respectively. Quadruple pentamers were occasionally seen. The self‐aggregation of human SAP molecules was investigated in the presence and absence of calcium ions at different concentrations. In calcium‐free solutions few and mostly small SAP aggregates were seen. After addition of calcium at increasing concentration the aggregates grew in size and crystallinelike structures were formed already at 2 mM calcium. At 25 mM calcium, large aggregates with a crystalline array occasionally exhibiting cylinders predominated. Binding of the ligands heparin and heparan sulphate to SAP completely abolished the calcium‐enhanced aggregation, but the distribution of the SAP molecules was affected, resulting in strands or groups of adjacent molecules. The electrophoretic mobility of SAP was moreover significantly altered after its calcium‐dependent reaction with these ligands. We conclude that purified SAP has a tendency to double pentamer formation and self‐aggregation also in the absence of calcium ions. However, aggregation is greatly enhanced even at low concentrations (2 mM) of calcium. SAP's tendency to self‐aggregation is abolished after its binding to heparin or heparan sulphate. Furthermore, our TEM studies indicate that purified human SAP freed of its natural ligands has the double pentameric form, whereas the electrophoretic investigations suggest that SAP's interaction with low‐molecular‐weight natural ligands in serum prevents homodimerization and self‐aggregation.
Effect of serum from HIV‐infected subjects on superoxide production by polymorphonuclear neutrophils
Authors: Pieri ELSA , Orsilles MIGUEL A.
Published: August 2009

Superoxide (O2−) production by polymorphonuclear neutrophils (PMN) was assessed in 17 subjects with asymptomatic infection (AI), 16 patients with acquired immunodeficiency syndrome (AIDS), and 12 healthy normal subjects. The effect of patients' serum on the oxidative activity of normal and patients' PMN was also investigated. The O2− production, nonstimulated and stimulated with zymosan particles in the presence of normal serum, was similar to that of normal controls in both groups of patients. The serum from 11 out of the 17 AI subjects (65%) induced an increase in the stimulated O2− production in normal and patients' PMN, while the serum from 8 out of the 16 AIDS patients (50%) induced a diminution. These effects did not appear to be related to complement C3 and circulating immune complex levels, but suggest that PMN of HIV‐seropositive patients do not present an intrinsic dysfunction and that the impact of serum factor(s) affects the normal oxidative activity of these cells depending on the stage of infection.
Carcinoembryonic antigen is expressed on endothelial cells
Authors: Majuri MARJA‐LEENA , Hakkarainen MERJA , Paavonen TIMO , Renkonen RISTO
Published: August 2009

Carcinoembryonic antigen (CEA) is a highly glycosylated cell surface protein. It is produced in large amounts in essentially all colon and several other adenocarcinomas. It has therefore been widely used as a clinical tumor marker. CEA is also a member of the immunoglobulin superfamily. Members of this family, such as ICAM‐1, ICAM‐2, VCAM‐1 and NCAM, are known to participate in cell‐cell adhesion. Similarly, the intercellular adhesion properties of CEA have been documented: it has been shown to mediate homotypic adhesion of cultured human colon adenocarcinoma cell lines. In this study we show for the first time that CEA is expressed on cultured human umbilical vein endothelial cells and on the endothelial cell line Ea.hy926. The expression of CEA on cultured endothelial cells can be enhanced by TNF‐α or IFN‐γ, and decreased by IL‐4. We demonstrate using immunohisto‐chemistry that anti‐CEA monoclonal antibody reacted with FVIII‐positive endothelium in tissue sections prepared from lymph nodes. Finally, we were able to show that CEA‐positive breast carcinoma cells bind to purified CEA protein, whereas CEA‐negative breast carcinoma cells do not. These results reveal for the first time that endothelial cells express CEA on the cell surface and suggest that CEA‐expressing adenocarcinomas could adhere to endothelial cells via CEA‐CEA interaction, thus facilitating tumor cell extravasation and hematogenic metastasis.
Antigen‐specific suppression of antibody responses by T lymphocytes cytotoxic for antigen‐presenting cells
Authors: Kagaya KEIKO , Shang HONG , Miyakawa YOZO , Fukazawa YOSHIMURA
Published: August 2009

In this study we demonstrated an alternative model of the antigen (Ag)‐specific suppression of antibody response in mice. Splenocytes that were taken from BALB/c mice immunized by i.v. injection of soluble human serum albumin (HSA) or ovalbumin exhibited MHC‐restricted Ag‐specific cytotoxicity for the respective antigen‐presenting cells (APC). When HSA‐primed splenocytes cultured with Ag and interleukin‐2 (IL‐2) were treated with anti‐CD4 or anti‐CD8 monoclonal antibody (mAb) plus complement, CD8+ and CD4+ T cells exhibited nearly the same level of cytotoxicity against APC. Furthermore, HSA‐primed CD4+ and CD8+ T cells released the same amount of interferon‐γ (IFN‐γ) when stimulated with Ag and IL‐2. Recombinant IFN‐γ was shown to suppress the in vitro plaque‐forming cell (PFC) response to sheep red blood cells (SRBC) only when it was added within 24 h after addition of Ag. The supernatants from both HSA‐primed CD4+ and CD8+ T cells suppressed the PFC response to SRBC in vitro, and the suppressive activity was abrogated by anti‐IFN‐γ mAb, but increased by anti‐IL 4 mAb. These results suggest that in our system the effector cells for Ag‐specific suppression of the antibody response in mice are both the cytotoxic type 1 clones (IFN‐γ‐producing) of CD4+ and CD8+ T cells for APC, and that IFN‐γ is a major extracellular effector molecule for such suppression.
Evaluation of the E‐test for susceptibility testing of the Bacteroides fragilis group
Authors: Bremmelgaard A. , Jansen J. E. , Justesen T. , Gottschau A. ,   Danish study group
Published: August 2009

The susceptibilities of 100 clinical isolates belonging to the Bacteroides fragilis group to 9 antibiotics, i.e. ampicillin, piperacillin, ceftriaxone, cefotaxime, cefoxitin, imipenem, erythromycin, clindamycin, and metronidazole, were tested using a standard agar dilution method and the E‐test. Overall, 81% of the E‐test MICs were within one log2 dilution step of the agar dilution MICs and 95% were within two log2 dilution steps. The E‐test showed significantly lower MIC values than the agar dilution method for ampicillin, cefotaxime, and imipenem, and significantly higher MIC values for piperacillin, ceftriaxone, erythromycin, clindamycin, and metronidazole. However, this only resulted in minor discrepancies that did not change the susceptibility status. The E‐test is easy to perform and read, and the MIC values correlated well with the MICs obtained by the agar dilution method when testing susceptibility of the Bacteroides fragilis group.
Mycotic and algal bovine mastitis in Denmark
Authors: AalbæK BENT , Stenderup JØRGEN , Jensen HENRIK ELVANG , Valbak JESPER , Nylin BRITTA , Huda ANNA
Published: August 2009

A one‐year examination of mammary secretions (n = 2,896) from Danish cattle with clinical or subclinical mastitis revealed 45 strains of fungi and algae. The strains originated from 44 mammary secretions of 42 cows in 40 herds. The following species of fungi were identified: Candida catenulata (n = 2), Candida kefyr (n = 6), Candida krusei (n=17), Candida rugosa (n = 6), Candida tropicalis (n = 3), Candida valida (n=1), Geotrichum capitatum (n = 5). The algal species Prototheca zopfii was demonstrated in five samples.