bluuebunny/crossref_metadata_2025 huggingface dataset:
Antigenic structure of the herpes simplex virus type 1 glycoprotein C: Demonstration of a linear epitope situated in an environment of highly conformation‐dependent epitopes
Authors: SJÖBLOM INGER , GLORIOSO JOSEPH C. , SJÖGREN‐JANSSON EVA , OLOFSSON SIGVARD
Published: August 2009

A continuous epitope, situated within or in close proximity to antigenic site II of the herpes simplex virus type 1‐specified glycoprotein C (gC‐1), was identified. The continuous linear nature of the epitope, defined by a monoclonal antibody C2H12, was establised by three independent lines of evidence: (i) The epitope was detectable by immunoblot under denaturing and reducing conditions, (ii) The epitope was detectable by RIPA of extracts from TM‐treated HSV‐infected cells, despite the malfolding caused by this treatment, (iii) The epitope was detected in an approximately 5,000‐dalton papain fragment of gC‐1. A mapping analysis, primarily based on use of mutant virus, expressing truncated gC‐1 molecules, suggested that the mapping position of the epitope was delimited by amino acids 120 and 230. Other epitopes of this region of gC‐1 are highly conformation‐dependent, and the existence of a linear epitope, accessible on native gC‐1, may facilitate the elucidation of the functional anatomy of gC‐1.
Coagulase‐negative staphylococci in a newborn with a cardiac vascular prosthesis
Authors: RYDING JAN , ESPERSEN FRANK , JARLØV JENS OTTO , ROSDAHL VIBEKE THAMDRUP
Published: August 2009

A case of septicemia with coagulase‐negative staphylococci in a newborn with a vascular graft is discussed. Nineteen different isolates of coagulase‐negative staphylococci were isolated from four different blood cultures. Eleven of the isolates belonged to one strain, which showed slight variation; the remaining seven isolates belonged to six different strains. The patient was treated for a prolonged period with antibiotics and seemed to respond. The diagnosis and proper treatment are discussed.
Antibodies against neutrophil cytoplasmic components in Kawasaki disease
Authors: SOPPI E. , SALO E. , PELKONEN P.
Published: August 2009

The occurrence of antibodies against neutrophil cytoplasmic components in 39 children (23 boys, 16 girls, median age 2.0 years) with Kawasaki syndrome was studied. The conventional indirect immunofluorescence test (ANC‐Ab) on alcohol‐fixed neutrophils and two commercially available ELISA tests (ANCA‐EIA and MPO‐EIA) were employed to detect the antibodies. Fourteen (36%) of the 39 patients with Kawasaki disease had antibodies against neutrophil cytoplasmic components in at least one of the three tests used. Eleven patients were identified using the indirect immunofluorescence test. Five patients were positive in the MPO‐EIA test and two additional patients in the ANCA‐EIA test. The IF staining pattern was cytoplasmic in eight patients and perinuclear in three. The cytoplasmic staining pattern in patients with acute Kawasaki disease is different from that seen in patients with Wegener's granulomatosis. The occurrence of antibodies may assist in the diagnosis of some patients with Kawasaki disease, although neither the positivity itself nor the five different antibody profiles seem to have any other clinical relevance.
Pathology of the heart in AIDS
Authors: HANSEN BIRGIT FISCHER
Published: August 2009

Cardiac disease and cardiac death in AIDS patients is seldom reported. In recent years minor cardiac abnormalities have been demonstrated, especially by echocardiography. Cardiac pathology in AIDS patients is here reported from 60 consecutive autopsies where the heart was investigated either using single samples of ventricular myocardium (the first 21 cases) or by an examination of the whole heart (the last 39 cases). Myocarditis according to the Dallas criteria was seen in 25 of 60 cases (42%), and in seven of these cases a probable pathogen (Toxoplasma gondii, cytomegalovirus, fungi) was demonstrated. Diffuse myocardial fibrosis was seen in 40 of 60 cases (67%) and is considered to be partly due to repair after myocyte necrosis/myocarditis. A myocardium thus weakened might not be able to meet an increase in functional demand, and in 15 of the 39 cases (38%) where an examination of the whole heart was performed, there was dilation and/or hypertrophy of the right ventricle. This is in agreement with our knowledge that the main diseases and main causes of death in AIDS patients are pulmonary. Survival time in AIDS is increasing due to ever improving symptomatic treatment, and the results of this study indicate that the prevalence of especially right‐sided heart failure will increase.
The prevalence of human papillomavirus in cervical lesions with koilocytosis and/or CINI
Authors: IVERSEN A. K. N. , DUUN S. , SEBBELOV A. M. , NORRILD B.
Published: August 2009

Thirthy‐one patients with koilocytosis and/or concomitant CIN I were analysed for the presence of HPV types 11, 16 and 18 by in situ hybridization and Southern blot analysis. The prevalence of HPV was 48% and 55%, respectively, when measured by the two methods and among the HPV positive, HPV 11 and 16 were present in 47% and 60%, respectively, whereas HPV 18 was not found.
Structure and function of L‐selectin
Authors: KANSAS GEOFFREY S.
Published: August 2009

The selectins are a newly described family of carbohydrate‐binding adhesion molecules involved in the regulation of leukocyte traffic. Selectins are composed of an N‐terminal C‐type lectin domain, a single EGF domain, a variable number of short concensus repeat (SCR) domains, a transmembrane region and a cytoplasmic tail. L‐selectin (LAM‐1/LECAM‐1/LECCAM‐1) is the only selectin expressed on leukocytes, and mediates a number of leukocyte‐endothelial interactions, including the binding of lymphocytes to HEV of peripheral lymph node high endothelial venules (HEV), neutrophil rolling, and leukocyte attachment to cytokine‐treated endothelium in vitro. Stable transfectants expressing a series of chimeric selectins and deletion mutants were functionally analyzed in order to determine the molecular basis of adhesion mediated by L‐selectin. The specificity of adhesion was found to reside entirely within the lectin domain, suggesting that this domain is the only domain of the protein to interact with the carbohydrate ligand. These results make previous observations that certain mAbs which block function map to each of the extracellular domains difficult to interpret. In addition, deletion of the cytoplasmic tail of L‐selectin abolished adhesion, without affecting ligand recognition. Thus, each domain of the selectins has an important, but distinct, role in cell adhesion.
Detection of human papillomavirus (HPV) using dot blot and Southern blot, hybridizing with a mixture of seven probes
Authors: LINDH ELISABET , CHUA KENG‐LING , KATAOKA AKIO , BISTOLETTI PETER , GROFF DENNIS , HJERPE ANDERS
Published: August 2009

A previously presented method for the detection and typing of human papillomavirus (HPV) DNA has been modified for the simultaneous analysis of HPV types 6, 11, 16, 18, 31, 33 and 35. The method has two steps, where a dot blot test is used to exclude cases that do not hybridize with HPV‐DNA. The remaining cases are then analysed by a Southern blot procedure, using a mixture of subgenomic probes for the simple and accurate analysis of HPV types. When the procedure was used for the analysis of clinical samples, patient groups at varying risk were found to differ with regard to the prevalence of HPV infections. Thus, the virus was detected in only 8.8% of otherwise healthy young women ‐ i.e. women without clinical signs of HPV‐related disease ‐ as compared to 48% of women who also had cervical intraepithelial neoplasia (CIN). A similarly high prevalence was found in patients in whom CIN persisted (42%) as compared to those in whom the morphological lesion regressed to normal (8%).
A distinct stratum corneum antigen in psoriasis and its reactions with stratum corneum autoantibodies
Authors: QUTAISHAT SALAH S. , KUMAR VIJAY , BEUTNER ERNST H. , JABLONSKA STEFANIA
Published: August 2009

Stratum corneum antibodies are ubiquitous and can be detected by various immunological methods. Of these, the ones detected by hemagglutination undergo changes in antibody titers and have been implicated in psoriasis. The purpose of our study was to examine if differences exist in the activities of the antigens isolated from psoriatic scales in comparison to normal callus. Stratum corneum antigens were prepared by trypsin‐phenol‐water extraction. The water phase, which contains the stratum corneum antigen, was used to sensitize the red blood cells in the hemagglutination assay. The antibody activity in human sera was determined before and after absorption with antigens isolated from callus, psoriatic scales, and cell envelopes. We found notable differences in the antigens obtained from callus and psoriatic scales. These include higher antibody titers to the antigens of the scales, the presence of unique antigenic determinants on psoriatic scales and the localization of the antigen on cell envelopes. These immunological differences were corroborated by the marked biochemical differences of certain amino acids, most notably glycine and proline, and these differences were unique to psoriatic scales as they were not shared with other hyperproliferative disorders.
Reduced rate of uptake of zinc ions in a calf affected with the lethal syndrome A46 relative to clinically normal calves using whole body radio‐isotope scanning of 62Zn
Authors: BAUER ROGERT , BRUMMERSTEDT ERIK , JENSEN MIKAEL , MEJBORN HEDDIE , SMITH MOGENS
Published: August 2009

Four clinically healthy control calves were scanned and blood clearance was determined from blood samples taken during the first hour after intravenous injection of 62Zn. Furthermore, one clinically healthy control calf and one calf suffering from the lethal hereditary syndrome A46 were scanned and blood samples taken from five min after intravenous and abomasal injection of 62Zn and onwards for a period of up to 26th. The results show a very fast liver uptake of 62Zn. The estimated rate of plasma/blood clearance and liver uptake for the control calves agrees with values derived from studies on other animals. In contrast, the A46 calf had a decreased rate of plasma/blood clearance and liver uptake relative to the control calves.
Adherence of Streptococcus pyogenes M type 5 to pharyngeal and buccal cells of patients with rheumatic fever and rheumatic heart disease during a one‐year follow‐up
Authors: KUMAR K. S. NANDA , GANGULY NIRMAL K. , ANAND INDER S. , WAHI PURSHOTAM L.
Published: August 2009

In vitro adherence of Streptococcus pyogenes M type 5 to isolated pharyngeal and buccal epithelial cells was studied in patients with acute recurrent rheumatic fever (n = 21), chronic rheumatic heart disease (n = 33), streptococcal pharyngitis (n = 12), and in normal controls. Patients were investigated at admission and one, six and 12 months later. Streptococci adhered significantly more to the pharyngeal cells of patients with rheumatic fever and rheumatic heart disease than to the pharyngeal cells of controls. Adherence of streptococci to pharyngeal cells of patients with pharyngitis was not different from age‐matched controls. The adherence of streptococci to the pharyngeal cells of patients with acute rheumatic fever fell during follow‐up but even after one year remained significantly higher than in the control group. These findings suggest that host factor(s) controlling streptococcal adhesion and colonization at the pharyngeal mucosa may be important in the pathogenesis of acute rheumatic fever.
Vascular rejection in cardiac transplantation
Authors: FOERSTER ARNOLD
Published: August 2009

Between November 1983 and January 1990 103 orthotopic heart transplants were performed at the National Hospital, University of Oslo, Norway. Twenty‐two patients died. Acute and/or chronic rejection was the cause of death in nine, disseminated infection in eight, cancer in three, and cerebral haemorrhage in one of the patients. Two patients were successfully retransplanted after graft failure due to ABO blood group incompatibility because of a communication error. One patient with a positive lymphocytotoxic crossmatch died shortly posttransplant due to acute circulatory collapse. The cumulative one‐ and five‐year survivals were 82% and 68%. Follow‐up time was 226.6 graft years, survival range from one to 2,306 days (mean 803 ± 4.12 SEM). A total of 1,343 endomyocardial biopsies were performed, which revealed 181 acute cellular rejection episodes, and 22 biopsies revealed acute and/or chronic vascular rejection. Autopsy studies showed three general types of vascular damage: acute (necrotizing) vasculitis, atherosclerotic disease in the epicardial arteries and diffuse proliferative arteriopathy in the intramural and smaller branches. In several cases acute vasculitis was concomitant with either of the two chronic types of accelerated graft sclerosis. Tissue immunofluorescence analysis demonstrated vascular deposition of immunoglobulin and complement in acute vasculitis, indicative of humoral immunoreaction. Postoperatively early chronic vascular rejection may develop.
Chronic gastritis and the distribution of parietal cells in duodenal ulcer patients and normals
Authors: BECK HENNING
Published: August 2009

Pentagastrin‐stimulated acid secretion and parietal cell density were studied in 51 patients with duodenal ulcer (DU) and 26 patients without ulcer disease (non‐DU). Chronic gastritis was assessed for a comparative study between parietal cell density and grade as well as the extent of gastritis. Parietal cell density was estimated with a high degree of accuracy, since the difference in cell counts of two adjacent biopsies and the inter‐ and intra‐observer variation never exceeded 10%. The density calculated from three regional biopsies was representative for the density of the entire corpus mucosa. Parietal cell density was similar in normal mucosa, superficial chronic gastritis and mild atrophic gastritis, while fewer parietal cells were found in severe chronic atrophic gastritis. Nonuniform distribution of chronic gastritis was more frequent in DU than in non‐DU, and may explain the lack of correlation between acid secretion and parietal cell number. If severe chronic atrophic gastritis is found to be localized, parietal cell density investigations will require multiple biopsies.
Vaginal leukocyte characteristics in urogenital trichomoniasis
Authors: BUCHVALD DUŜAN , DEMEŜ PAVOL , GOMBOŜOVÁ ANNA , MRÁZ PETER , VALENT MICHAL , ŜTEFANOVIĈ JÁN
Published: August 2009

Morphological and functional characteristics of vaginal exudate leukocytes were examined in 47 patients with urogenital trichomoniasis. Electron microscopic morphology, viability, phagocytosis of Candida albicans blastospores and ability to undergo respiratory burst in the iodonitrotetrazolium reductase test were evaluated in these cells. Vaginal inflammatory leukocytes were almost exclusively polymorphonuclear neutrophils, and their concentration was positively correlated (r = 0.58; p<0.001) with the number of trichomonads in the exudate. Median leukocyte viability reached 39% and both phagocytic and tetrazolium reductase activities of these cells were significantly reduced in comparison with those of circulating polymorphonuclear leukocytes. Patients with a clinical picture of severe mucosal inflammation had significantly higher vaginal exudate leukocyte concentrations and viability than those without inflammatory signs. The possible role of vaginal leukocytes in the pathogenesis of urogenital trichomoniasis is discussed.
Effect of anti‐interleukin‐1 administration to rats with adriamycin‐induced nephrosis
Authors: BRICIO TERESA , MOLINA ANA , MAMPASO FRANCISCO
Published: August 2009

Rats receiving a single dose of adriamycin (7.5 mg/kg) develop heavy proteinuria and histologic lesions similar to those found in minimal change nephrotic syndrome in humans. We found that whole isolated glomeruli from rats injected with adriamycin secreted an IL‐1‐like cytokine which closely resembled macrophage IL‐1. Maximal IL‐1‐like activity was detected on day 14 of the experiment when rats were heavily proteinuric. Administration of anti‐IL‐1 antiserum to rats with adriamycin‐induced nephrosis provoked a transient but marked reduction in the urinary protein excretion. Our results indicate that IL‐1 could be an important mediator implicated in the development of proteinuria in this experimental nephropathy.
Altered expression of ABO(H) carbohydrate antigens is seen in pleomorphic adenomas
Authors: THERKILDSEN MARIANNE HAMILTON , MANDEL ULLA , CHRISTENSEN MARIA , BARFOED CLAUS , DABELSTEEN ERIK
Published: August 2009

Cell surface carbohydrate antigens show changes in relation to differentiation, maturation and malignant transformation. The expression of type 2 chain ABH carbohydrate structures of the ABO histo‐blood group system was investigated in 28 pleomorphic adenomas (PA) and normal parotid glands in order to study possible changes in the glycosylation pattern. The distribution of carbohydrate structures was investigated by immunohistological stainings of formalin‐fixed paraffin‐embedded material using monoclonal antibodies (MAbs) with well‐defined specificity. A strong interindividual variation was found in the normal tissue as well as in the tumors. In normal tissue, acinus and duct cells all expressed elongated carbohydrate structures. The myoepithelial cells did not stain with any of the MAbs investigated. In the PAs, staining was seen in the ductular structures and myoepithelial cells. In contrast to normal tissue, the tumors expressed the short precursor molecule sialylated N‐acetyllactosamine. Furthermore, the PAs showed loss of H and A antigens, and a reduced expression of Ley compared to normal tissue. The ductular structures as well as the modified myoepithelial cells expressed binary N‐acetyllactosamine, which in the normal tissue could only be found in the striated and excretory ducts. Thus our study has shown that aberrant glycosylation is not only a feature of malignant neoplasms but also occurs in pleomorphic adenomas.
Phagocyte killing of Campylobacter jejuni in relation to oxidative activation
Authors: WALAN ASA , DAHLGREN CLAES , KIHLSTRÖM ERIK , STENDAHL OLLE , LOCK RONNY
Published: August 2009

When studying the interaction between Campylobacter jejuni and human neutrophils, we found that four different clinical isolates showed a great variability in this association. Also the ability to induce neutrophil production of oxidative metabolites, measured as chemiluminescence (CL), differed between the strains. Surprisingly, strain 1, which showed weak interaction with neutrophils and high resistance to killing, induced the highest CL response. All strains evoked an intracellular CL response, and three strains also gave rise to an extracellular response. This extracellular release of toxic oxygen species might contribute to the local tissue damage during infection. There was no clear correlation between association, killing and oxidative response. However, one strain that only evoked an intracellular generation of oxygen metabolites also showed the highest sensitivity to killing. Phagocytosis was increased up to ten times after opsonization with normal human serum. The intracellular CL production increased several fold, whereas the extracellular generation of oxygen species disappeared or was considerably decreased after opsonization. These results indicate that complement‐opsonized C. jejuni are phagocytosed and readily attacked by the oxidative defence system within the phagosome.
Plasma proteins in normal neurons
Authors: LØBERG E. M. , TORVIK A.
Published: August 2009

Previous observations have shown that plasma proteins are taken up by damaged neurons after trauma, cold injury and anoxic lesions. The present study was undertaken to test whether such uptake also can occur in normal neurons. Normal human brains at different intervals after death as well as normal rat brains after perfusion and immersion fixation were studied. During the first 24 h after death, the human cases showed practically no immunostaining with anti‐fibrinogen and vague and inconstant staining with anti‐albumin in the cortex and hippocampus, while certain brain stem nuclei were stained with both antisera even at the shortest postmortem intervals. At longer intervals, increased immunostaining of the neurons in the cortex and hippocampus was also demonstrated. Generally, more cells were positive for albumin than for fibrinogen. Perfusion‐fixed rat brains showed no immunostaining, whereas rat brains that were immersion‐fixed 24 h after death showed positive staining, mainly located in the brain stem. It is concluded that the staining of normal neurons demonstrated with the present method is due to postmortem leakage of proteins. Since practically no uptake of fibrinogen occurs in the cortex or hippocampus during the first 24 h, a definite neuronal staining with anti‐fibrinogen at these intervals probably indicates true neuronal damage. In the brain stem, on the other hand, this phenomenon can not be used to demonstrate neuronal damage even at the shortest postmortem intervals.
Early development in healthy children of serum opsonins against nonpathogenic Neisseria meningitidis
Authors: FREDLUND HANS , SELLDÉN HANS , DANIELSSON DAN , OLCÉN PER
Published: August 2009

In an earlier study, with the use of chemiluminescence (CL) and phagocytic killing, we could show that in the presence of serum from healthy adults polymorphonuclear leukocytes (PMNL) efficiently handle nonpathogenic Neisseria meningitidis strains, in sharp contrast to those associated with clinical disease. The major part of this difference was dependent on serum factors. In the present study 84 serum samples from children 1–3, 4–6, 7–9, and 10–14 years old were studied by the CL technique according to their ability to opsonize meningococci. There was a highly significant difference (p < 0.001) in all four age groups when the CL indexes obtained with the pathogenic meningococci of the serogroups A, B and C were compared with those of the nonpathogenic menigococci: serogroup 29E and nongroupable meningococci. These findings imply that the ability to opsonize so‐called nonpathogenic meningococci is developed early in life and may explain why they are only occasionally able to cause disease.
Yersinia enterocolitica serotype 0:3‐induced arthritis in mice: microbiological and histopathological information
Authors: TOYOS JUAN R. DE LOS , MENÉNDEZ PRIMITIVA , SAMPEDRO ANDRÉS , HARDISSON CARLOS
Published: August 2009

Gross anatomical and histopathological changes in arthritic joints resulting from oral challenge with Yersinia enterocolitica serotype 0:3, upon pretreatment with desferrioxamine, were always more severe than those induced by intravenous infection of immunized animals. In all the acute inflammation episodes studied, live Yersiniae were isolated from the arthritic region. Invariably, a heavy mixed infiltration of synovia, joint spaces and soft tissues was observed at this stage. Concurrent fibrous thickening and vascular proliferation, along with erosion of articular cartilages and anomalous bone regeneration, were also apparent. In spite of these significant facts, the bacterium could be histopathologically identified only in bone marrow where it developed microcolonies and caused significant necrosis as well. The live bacterium was also retrieved from two‐ and six‐month‐old arthritic ankles/paws examined, but it could not be seen in histological sections of joints. By this time, no cellular infiltration was evident, but there was extensive fibrosis. Bones were at times greatly enlarged, showing a spongeous‐like structure. Additionally, articular cartilages could be completely lost and were substituted by an anomalous ossification filling the joint spaces. This situation led to bone fusion, resembling articular ankylosing traits. In summary, we present the first experimental evidence that Y. enterocolitica serotype 0:3 is a causal agent of osteoarthritis and osteomyelitis, and that it may survive for prolonged periods of time in osseous structures.
Interferon‐gamma may enhance infection of blood‐derived macrophages with HTV‐1 in the presence of HIV‐positive serum
Authors: DEGRÉ MIKLOS , BECK SOLVEIG , ROLLAG HALVOR
Published: August 2009

HIV multiplication in blood‐derived macrophages was slightly inhibited by pretreatment of cells with interferon‐gamma or by preincubation of virus with serum containing antibodies against HIV. When these pretreatments were combined, the HIV titres observed a short time after infection were enhanced. This effect was blocked by antibodies against Fc receptors but not by antibodies against CD4 receptors. Interferon enhanced the expression of Fc receptors on macrophages. The results indicate that IFN‐γ, in appropriate combinations with HIV‐antibody‐containing human serum, may enhance the rate of HIV infection of macrophages.
Characterization of a 25,000‐dalton Helicobacter pylori protein, cross‐reacting with a Campylobacter jejuni protein
Authors: BUCHVALD DUŜAN , MÆLAND JOHAN A.
Published: August 2009

Extract obtained by ultrasonic disruption of Helicobacter pylori bacteria contained a protein with subunit molecular mass of 25 kD which bound antibodies in sera from patients with H. pylori associated disease. The protein was purified by gel permeation and elution from SDS‐polyacrylamide gel slices, and was used to raise an anti‐25‐kD protein‐specific rabbit serum. Using the antiserum in experiments, the results indicated the following: The protein exists as covalently linked dimers (45 kD) of the 25‐kD subunits. Variable numbers of non‐covalently linked copies of the dimers make up the native protein. The protein was susceptible to digestion by papain, pronase, and trypsin. Pepsin cleaved off a fragment of approximately 2 kD. A small share of the protein was exposed at the bacterial cell surface, the greatest share being localized internally. The protein was not secreted and it was probably not an integral part of the outer membrane. It was produced in variable quantity by all of 11 H. pylori strains tested and was a major protein in some strains. A cross‐reacting protein with subunit size of 25 kD was also produced by Campylobacter jejuni strains, but not by any of a variety of other bacteria. Since both H. pylori and C. jejuni infection occur with a high frequency, the cross‐reacting 25‐kD protein may interfere unfavourably with the diagnostic specificity of serological tests for infection caused by these bacteria.
Screening method for β‐lactamase substrate profiles
Authors: SCHUMACHER H. , NIR M. , MANSA B. , THOMSEN V. F.
Published: August 2009

A disc diffusion method, based on the idea of Klundert, for screening of substrate profiles of β‐lactamases was developed in order to perform epidemiological studies. The method was tested against 30 different reference β‐lactamases and 59 clinical isolates known to produce TEM‐1, SHV‐1 and BRO‐1. The reproducibility and discriminating power of the disc diffusion method made it possible to differentiate between eight types of substrate profiles for the 30 reference β‐lactamases and to differentiate between TEM‐1, SHV‐1 and BRO‐1 from clinical isolates. In combination with analytical isoelectric focusing the disc diffusion method gives a reliable identification of β‐lactamases.
Northern and Southern blot analysis of human RNA and DNA in autopsy material
Authors: LARSEN SUSANNE , RYGAARD KÅRE , ASNÆS STIG , SPANG‐THOMSEN MOGENS
Published: August 2009

Fresh biopsy material for molecular biological investigations is not obtainable from all relevant normal human tissues. We studied the feasibility of using RNA and DNA from autopsies for Northern and Southern blot analysis. Tissue samples from seven organs were obtained from 10 autopsies performed 21–118 h postmortem. Extracted RNA and DNA were examined by Northern and Southern blot analysis using oligolabelled human DNA probes recognizing gene transcripts of 2–5 kb. The results indicated that, in general, Northern blot analysis was feasible with the applied probes when the tissue was obtained less than two days postmortem. Histological examination showing slight or no autolysis and the presence of ribosomal bands after gel electrophoresis were both indicative parameters of RNA preservation. DNA was appropriate for Southern blotting when the tissue was obtained less than three to five days postmortem, depending on the organ from which the DNA was extracted.
Use of o‐phthalaldehyde to detect O‐phosphorylethanolamine in bacterial lipopolysaccharide
Authors: YEH HUNG‐YUEH , PRICE RAY M. , JACOBS DIANE M.
Published: August 2009

We have developed a method to measure O‐phosphorylethanolamine groups in bacterial lipopolysaccharide using a fluorescent reagent, o‐phthalaldehyde. The optimal excitation and emission wavelengths were 335 nm and 450 nm, respectively. The reaction was pH‐dependent with an optimum at pH 10.5. The maximum fluorescence intensity occurred two min after mixing lipopolysaccharide with the reagent at pH 10.5. The assay was linear over a range of 1 μg to 100 μg of lipopolysaccharide. When we compared the amount of primary amine (as O‐phosphorylethanolamine) in native and p‐hydroxyphenylacetic acid‐derivatized lipopolysaccharide, we found that 97% of amine groups in native lipopolysaccharide were derivatized by p‐hydroxyphenylacetic acid in the presence of 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide.
An ultrastructural and cytochemical study of the pulmonary lesions and multinucleate giant cells in porcine dermatosis vegetans
Authors: EVENSEN ØYSTEIN , BRATBERG BJØRN
Published: August 2009

The pulmonary lesions and multinucleate giant cell formations in porcine dermatosis vegetans were studied ultrastructurally and cytochemically in seven affected pigs that ranged from one to 120 days of age. At birth, no pulmonary lesions were observed. By seven days of age, there was a moderate infiltration of monocytes/macrophages in the alveoli and interstitium. These changes progressed, and by two weeks of age there was pronounced infiltration of macrophages and lymphocytes in the alveoli. Close apposition of macrophages and lymphocytes was observed, and cellular contact was demonstrated. By three to four weeks of age, small aggregates with epithelioid cells, macrophages, and lymphocytes resembling granulomas were present. In an older pig, aged four months, the inflammatory changes waned. Large macrophages, and large and small multinucleate cells shared common morphological characteristics of cytoplasm endowed with organelles, primary and secondary lysosomes, Golgi profiles, and granular endoplasmic reticulum. Cytochemically, macrophages and MGCs were positive for acid phosphatase. The present study showed that the initial pulmonary changes share morphological characteristics with a granulomatous inflammatory response, and evolve into small granulomas with macrophages, epithelioid cells and lymphocytes. Morphologically, these changes share characteristics with epithelioid cell granulomas. Macrophages and MGCs share common ultrastructural traits, and are positive for acid phosphatase. MGCs seem to evolve in the course of the granulomatous inflammatory response and are probably of monocyte/macrophage origin.
Inducible beta‐lactamases in clinical isolates of non‐aeruginosa Pseudomonas
Authors: RAVAOARINORO MADELEINE , TOMA EMIL , FALLARA ALAIN
Published: August 2009

The incidence of antimicrobial resistance and expression of imipenem‐inducible beta‐lactamase were examined in 22 strains of non‐aeruginosa Pseudomonas isolated from clinical specimens. The percentage of strains resistant to from one to eight antibiotics was 45. The most active antibiotics against all strains were norfloxacin, ciprofloxatin and imipenem. Eighteen out of the 22 strains were positive for beta‐lactamase in a spectrophotometric assay using nitrocefin as substrate. A low inducible beta‐lactamase specific activity (0.001‐0.999 nmoles nitrocefin hydrolyzed/min/mg protein) was found in twelve strains whereas six strains had a relatively high specific activity (3.5–159.8 nmoles nitrocefin hydrolyzed/min/mg protein). Five strains representing different Pseudomonas spp. and showing high beta‐lactamase activity were studied further. Crude enzymes from two species (Pseudomonas mendocina, Pseudomonas acidovorans) hydrolyzed cefazolin at a higher rate than penicillin and ampicillin. All enzymes from the five species were inhibited by cloxacillin and p‐chloromercuribenzoate (1 mM), but were insensitive to inhibition by clavulanic acid, ethylenediamine acetic acid (EDTA) at the same concentration. The isoelectric point and molecular weight of the main beta‐lactamase band from the 5 species were 6.5–6.8 and 47000 respectively.
Ultrastructural study of the response of cells infected in vitro with causative agent of spotted fever group rickettsiosis in Japan
Authors: IWAMASA KIKUE , OKADA TAKANORI , TANGE YOSHIKI , KOBAYASHI YUZURU
Published: August 2009

The response of host cells L929 infected with causative agent of spotted fever group (SFG) rickettsiosis in Japan, the Katayama strain, was studied by electron microscopy. The rickettsiae penetrated the cytoplasm and multiplied here and after prolonged incubation progressed into the dilated cisternae of rough endoplasmic reticulum (rER), the perinuclear space, the deep invaginated nuclear membrane, and then the nucleoplasm of the host cells. The intranuclear rickettsiae showed different states: one type was enclosed by the double membrane of the host cell and the other type was free in the nucleoplasm. In addition to these double membrane‐bound and membrane‐free intranuclear rickettsiae, various membrane structures, including rER‐like structures, were also found in the nucleus. The cells infected with the rickettsiae underwent distinctive morphological alterations which occurred mainly within intracellular membranes of the host cells. These findings indicate the possibility that the intracellular membranes are characteristic cytopathological sites in rickettsia‐host cell interaction, and that these alterations may be related to a possible route of rickettsial penetration into the nucleus: passage through vesicles formed from invaginations in the nuclear membrane.
In vitro activity of teicoplanin and vancomycin against Grampositive bacteria from human clinical and veterinary sources
Authors: JENSEN KJELD TRUBERG , SCHØNHEYDER HENRIK , PERS CHARLOTTE , THOMSEN VILLY FRØLUND
Published: August 2009

The minimum inhibitory concentration (MIC) of teicoplanin and vancomycin was determined by the agar dilution method for 186 Gram‐positive bacteria from human clinical and veterinary sources. Teicoplanin MIC values were ≤4 μg/ml for 94% of staphylococci (group A, n = 52) and ≤2 μg/ml for all streptococci, enterococci, aerococci and pediococci (group B, n = 75). Seventy‐eight percent of Gram‐positive rods, Rhodococcus and Leuconostoc spp. (group C, n = 59) were inhibited by 4 ug/ml. Teicoplanin resistance (MIC ≥ 16 μg/ml) was demonstrated for all Nocardia strains and for some strains of Lactobacillus, E. rhusiopathiae, Leuconostoc, and S. haemolyticus. Cross‐resistance between teicoplanin and vancomycin was observed for all Nocardia strains and for some strains of Lactobacillus, E. rhusiopathiae, and Leuconostoc. Three methicillin‐resistant S. haemolyticus strains were either resistant or intermediately susceptible to teicoplanin and susceptible to vancomycin. Eight strains (motile enterococci four, E. rhusiopathiae three and Leuconostoc sp. one) were susceptible to teicoplanin and resistant to vancomycin. Teicoplanin disc diffusion on Danish Blood Agar with NeoSensitabs® (Rosco), PDM® AB Biodisc and locally prepared discs revealed a wide range of zone diameters in groups B and C. The relation between MIC values and zone diameters for teicoplanin was analysed by the error‐rate bounded method. Zone size interpretive criteria as suggested by the manufacturers (≥ 15 mm) produced 2.7% (95% confidence limits 0.9‐6.2%) and 1.6% (95% confidence limits 0.3–4.6%) very major errors for NeoSensitabs and PDM‐disc, respectively. Using a zone size breakpoint for susceptibility of ≥ 25 mm for NeoSensitabs and ≥20 mm for PDM‐disc, the proportions of very major errors were 0.5% (95% confidence limits 0.0–3.0%) at the expense of 5.9% (95% confidence limits 3.0–10.3%) indeterminate strains that belonged to E. rhusiopathiae, Leuconostoc, Lactobacillus and S. haemolyticus. However, using these zone size breakpoints five major errors (β‐haemolytic streptococci, group B three, S. aureus one, Leuconostoc sp. one) were observed for NeoSensitabs and two major errors (β‐haemolytic streptococcus, group B one, Leuconostoc sp. one) were observed for PDM‐disc. Susceptibility testing against teicoplanin among these taxa should therefore include a determination of MIC.
Dietary effects of barley fibre, wheat bran and rye bran on bile composition and gallstone formation in hamsters
Authors: ZHANG JIE‐XIAN , LUNDIN EVA , HALLMANS GÖRAN , BERGMAN FRANK , WESTERLUND ERIC , PETTERSON PETER
Published: August 2009

The effects of brewer's spent grain (BSG), wheat bran and rye bran on bile composition, gallstone formation and serum cholesterol were studied in Syrian golden hamsters. The frequency of gallstone formation in the animals fed diets supplemented with low (10%) and high (20%) concentrations of BSG or wheat bran was significantly lower than that of the animals fed a stone‐provoking, fibre‐free diet. The ratios of secondary to primary bile acids were lower in the animals fed the diets supplemented with a high dose of BSG, wheat bran and rye bran than in the controls. The ratio of LCA to DCA was reduced only in the animals fed the diet supplemented with rye bran as compared with controls. No significant changes in bile and serum cholesterol levels were observed in the experiment.
Tumours classified as “malignant histiocytosis” in children are T‐cell neoplasms
Authors: ORNVOLD KIM , CARSTENSEN HENRIK , JUNGE JETTE , GYHRS ANETTE , RALFKIAER ELISABETH
Published: August 2009

During the last five years increasing evidence has accumulated that many tumours classified as ‘histiocytic’ in the past do not originate from macrophages, but from transformed (or anaplastic) large lymphoid cells. Most of these studies have focused upon adult neoplasms. Knowledge concerning the lineage of ‘histiocytic’ tumours in the paediatric age group is more limited. In this study we have examined the clinical, morphological and immunophenotypical features of six childhood malignancies originally diagnosed as being of histiocytic origin. Three patients showed an aggressive course with involvement of internal organs and very short survival times. Two patients were brought into remission: one is alive without active disease after seven years; the other died after seven years due to treatment‐related cardiomyopathy. The remaining patient had a protracted course for two and a half years, but subsequently deteriorated and died three years after diagnosis. The histomorphological features in five cases were those of anaplastic large cell lymphomas. The remaining case consisted of pleomorphic (rather than anaplastic) large lymphoid cells. In all cases the immunophenotypical examination showed features characteristic of activated T lymphocytes. All cases were positive for Ki‐1 (CD30), and three were positive for epithelial membrane antigen (EMA). Histiocyte‐associated markers were positive in residual reactive macrophages, but nowhere could unequivocal positivity for macrophage‐associated markers be seen in the neoplastic cells. It is concluded that most childhood malignancies in the past classified as ‘histiocytic’ are examples of anaplastic large cell (Ki‐1) lymphomas of T‐cell type and that true histiocytic malignancies are exceedingly rare in the paediatric age group.
Role of streptococcal IgG Fc receptor in tissue deposition of IgG in rabbits immunized with Streptococcus pyogenes
Authors: BUROVA LARISSA A. , KOROLEVA IRINA V. , OGURTZOV ROBERT P. , MURASHOV SERGEJ V. , SVENSSON MAJ‐LIS , SCHALÉN CLAES
Published: August 2009

Induction of anti‐IgG during hyperimmunization of rabbit with Streptococcus pyogenes (group A streptococci; GAS) was previously shown to require the presence of IgG Fc receptors (FcR) in the vaccine strain. In the present work, we examined whether streptococcal FcR activity might also be of importance for heart and kidney deposition of IgG, known to occur in poststreptococcal sequelae as well as during experimental immunization of animals. Each of three IgG‐binding (GAS types M1, M12 and M22) and two non‐binding (GAS type T27 and S. agalactiae (GBS) type Ia) streptococcal strains were used for intravenous immunization of rabbits during two periods of eight and six weeks, respectively, separated by an interval of one month. Before use, vaccine strains were treated with KSCN and carefully washed in order to remove any surface‐bound immunoglobulins. No deaths occurred among injected rabbits. No tissue deposition was elicited by the GAS type T27 or the GBS strain. In contrast, the strains of types M1, M12 and M22 all induced deposits of IgG in kidney and heart tissue, beginning during the first immunization period. In two tested animals, receiving GAS of types M1 or M22, circulating immune complexes containing anti‐IgG antibodies were also detected. Finally, serum autoantibodies reacting with preparations of heart and kidney, but not lung or liver, were demonstrated in each of six animals receiving Ml or M22, reaching maximum levels during reimmunization; such antibodies were not evoked by the two strains not binding IgG. Our results suggest that, in GAS with capacity for non‐immune binding of IgG, triggering of anti‐IgG acted to enhance tissue deposition of IgG or immune complexes in immunized rabbits. Furthermore tissue‐specific antibodies were elicited only by the IgG‐binding strains and occurred comparatively late during immunization, suggesting that those antibodies might have been triggered due to the exposition of hidden kidney and heart determinants.
Lysozyme localization in normal and diseased human gastric and colonic mucosa
Authors: SANTINI DONATELLA , PASQUINELLI GIANANDREA , MAZZOLENI GUIDO , GELLI MARIA CAROLINA , PREDA PAOLA , TAFFURELLI MARIO , MARRANO DOMENICO , MARTINELLI GIUSEPPE
Published: August 2009

The distribution of lysozyme in normal and pathological human gastric and colonic mucosa was studied by light and electron microscopic immunocytochemical techniques and compared with histological and histochemical features. Lysozyme was localized in pyloric glandular epithelial cells, mucous neck cells of fundic glands, Paneth cells and some crypt cells of the mature colonic mucosa. In addition, lysozyme was detected in a large spectrum of “immature” or “regenerative” epithelium: neck cells of the gastric regenerative zone, undifferentiated columnar cells of surface and hyperplastic interfoveolar crests of the stomach, regenerative cells in a healed gastric ulcer, some goblet cells in incomplete intestinal metaplasia, cells of the regenerative zone at the bottom of colonic crypts and, finally, fetal intestinal epithelium. Electron microscopically, we localized lysozyme in the central core of mucous granules in the pyloric gastric glandular epithelium and in the dense mucous granules in gastric mucous neck cells. Lysozyme was also detected in some immature mucin‐producing cells of the gastric regenerative zone and in the rough endoplasmic reticulum of surface hyperplastic columnar gastric cells. At the electron microscopic level, a peculiar correlation between the immunopattern of lysozyme and the morphology of mucous granules has been postulated. All our data support and extend the view that the presence of lysozyme may be related to cell immaturity as well as to a regenerative state of the cell. Finally, the lysozyme distribution and its relation to mucosubstances in gastric and colonic carcinoma suggest that lysozyme should not be considered an exclusive marker of cells of gastric derivation.
The redistribution of lymphocytes during adrenaline infusion
Authors: TOFT P. , TØNNESEN E. , SVENDSEN P. , RASMUSSEN J. W. , CHRISTENSEN N. J.
Published: August 2009

Adrenergic activation is known to occur in sepsis and after major surgery or trauma. An elevated serum concentration of adrenaline is followed by lymphocytosis in peripheral blood even in splenectomized patients. The purpose of the present study was to clarify the redistribution of lymphocytes in the tissues during adrenaline infusion. Lymphocytes were isolated from 24 rabbits, labelled with indium‐111‐tropolone and reinjected into the rabbits. The next day the rabbits were anaesthetized. Eight rabbits received 3 μg of adrenaline i.v. followed by 0.2 μg/min, eight received 300 μg of adrenaline i.v. followed by 20 μg/min, while eight received a saline infusion and served as a control group. The activity of labelled cells was imaged with a gamma camera and computer before, during and after adrenaline infusion. The activity of the spleen decreased to 90% and 94% of initial values during low and high doses of adrenaline. The activity of the bone marrow decreased to 91% and 96%, respectively, while the activity of the heart/lung and the liver increased to 107% and 106% with the high dose of adrenaline. In peripheral blood the lymphocytes increased 10%. It is concluded that lymphocytes are redistributed from spleen and bone marrow to peripheral blood, lungs and liver during adrenaline infusion in this animal model.
Aspergillus fumigatus fungaemia and myocarditis in a patient with acquired immunodeficiency syndrome
Authors: SCHØNHEYDER HENRIK , HOFFMANN STEEN , JENSEN HENRIK ELVANG , HANSEN BIRGIT FISCHER , FRANZMANN MARIA‐B.
Published: August 2009

Necrotizing myocarditis due to Aspergillus fumigatus was a contributory cause of death in a patient with acquired immunodeficiency syndrome and non‐Hodgkin lymphoblastic malignant lymphoma of the Burkitt type. A transient remission of the lymphoma had been obtained by cytostatic treatment. A. fumigatus was isolated from blood two weeks before death, but myocarditis was not diagnosed until autopsy.
Detection of mycobacteria from blood and bone marrow: a decade of experience
Authors: ASKGAARD DORTHE , FUURSTED KURT , GOTTSCHAU ADAM , BENNEDSEN JØRGEN
Published: August 2009

This study reports our experience with methods used at our department from 1981 through 1990 for detection of mycobacteria in blood and bone marrow specimens. Direct inoculation on Lowenstein Jensen media was replaced by Isolator® lysis‐centrifugation followed by inoculation on conventional solid media, and the Bactec 12B and Bactec 13A systems. A total of 3033 specimens were analyzed. A total of 137 mycobacterial isolates were obtained from 42 patients, all HIV‐positive except one. Mycobacteremia caused by M. avium‐intracellulare (83%), M. tuberculosis, M. scrofulaceum and M. kansasii was found. Of 680 blood specimens tested by the last three methods, 7.6% were found to be positive by at least one method and revealed recovery rates of 6.8% for the Isolator®‐solid media system, 3.4% for the Isolator®‐12B system and 6.9% for the 13A system (all isolates MOTT). Mean detection times for 21 cultures found positive by all three methods were 23.6, 23.3 and 17.7 days for the Isolator®‐solid media, Isolator®‐12B and 13A systems, respectively, with a significantly shorter detection time for the 13A system. Low degree (< 1 cfu/ml) mycobacteremia (MOTT) caused delay in the Isolator®‐solid media and the 13A systems and no detection in the Isolator®‐12B system. Antituberculous therapy significantly prolonged the detection times for MOTT in the 13A system in contrast to the other systems.