bluuebunny/crossref_metadata_2025 huggingface dataset:
Mouse cytolytic T cells reactive with rat islet tumour RIN 5AH‐B cells
Authors: SchØLler JØRGEN , Rubin BENT
Published: August 2009

Insulin dependent (type 1) diabetes mellitus is considered to be an autoimmune disease characterized by a specific destruction of the insulin‐producing pancreatic β cells. We have explored the possibilities of raising T cells specific for putative pancreatic β‐cell autoantigens using a xenogeneic system. Mouse T cells were induced against the rat insulinoma cells RIN 5AH‐B (RIN) and tested for their specific reactivity. No MHC class II‐restricted β‐cell‐specific helper T‐cell reactivity could be detected within the bulk cultures as measured by proliferation, but a remarkably high cytotoxicity against the RIN cells was observed. The target antigen on the cell surface recognized by the generated cytotoxic T cells was shown to be the rat class I major histocompatibility antigen RT18, and not a β‐cell or tumour cell‐specific antigen associated with RIN cell MHC molecules. Our results demonstrate that it is feasible to evoke a xenogeneic T‐cell response against the RIN cells. However, the mouse T cells recognize a dominant epitope present on the expressed rat class I major histocompatibility antigen RT1A8 and not a β‐cell‐specific antigen. Hence, we conclude that it appears most unlikely that β‐cell‐specific T cells can be raised in the xenogeneic system.
Urothelial metaplasia of the bladder mucosa
Authors: Kvist EBBE , SjØLin KNUD‐ERIK
Published: August 2009

Three different types of metaplasia of the urinary bladder are described: nephroid metaplasia, colonic metaplasia and signet‐ring cell metaplasia. The latter metaplasia was without any histological signs of malignancy and the patient is still alive without recurrence, 12 years after the diagnosis was made. We believe it is the first published case of benign signet‐ring cell metaplasia. The patients with nephroid metaplasia were controlled from 3–18 months without recurrence. Two of the patients with colonic metaplasia were followed from 16 to 132 months; both developed recurrences. In patients with metaplasia of the urothelium, follow‐up cystoscopy is recommended because of the recurrent nature of the condition and the malignant potential of metaplastic urothelium.
Orchitis and testicular abscess formation caused by non‐typhoid Salmonellosis
Authors: Ejlertsen T. , Jensen H. K.
Published: August 2009

The case of a 36‐year‐old man with epididymo‐orchitis followed by testicular abscess formation 4 weeks later due to Salmonella berta is reported, and the literature on orchitis and testicular abscesses caused by non‐typhoid Salmonella is reviewed.
Phage‐typing of coagulase‐negative staphylococci. Factors influencing typability
Authors: Rosdahl VIBEKE THAMDRUP , Gahrn‐Hansen BENTE , MØLler JENS K. , KjæLdgaard POUL
Published: August 2009

Factors influencing the phage‐typability of coagulase‐negative Micrococcaceae have been studied in 2,778 clinical isolates comprising A) 209 consecutive isolates from one laboratory, B) 2,107 clinical strains submitted for phage‐typing for epidemiological reasons, and C) 462 strains representing all isolates of presumed clinical significance found in two laboratories during one month. The reproducibility was acceptable at duplicate repeated typing of the same strains as well as by typing epidemiologically‐related pairs of strains from the same patient. Strains of Staphylococcus haemolyticus were seldom typable, whereas strains of S. epidermidis and S. hominis had a higher typability. Methicillin‐resistant strains and other multiple‐resistant strains were rarely typable (11–13%). The typability was higher among susceptible strains (36%) and strains resistant to penicillin only (43–50%). The typability of strains of the same species and antibiotic‐resistance pattern differed between hospitals compared and decreased markedly over the years for multiple‐resistant S. epidermidis isolates.
The influence of a histamine2‐receptor antagonist on the healing of an experimentally induced gastric mucosal lesion
Authors: Park PER‐OLA , Alumets JAN , Arvidsson STEFAN , Grimelius LARS , Haglund ULF
Published: August 2009

The effect of an H2‐receptor antagonist (ranitidine) on the healing of gastric mucosal lesions was studied. Mucosal lesions were induced by a standardized thermo‐mechanical technique. The healing process was assessed by macro‐ and light microscopical examination. It was further evaluated by measurements of the tissue contents of hydroxyproline, a chemical compound reflecting collagen, and of DNA and RNA, reflecting cell frequency and protein synthesis respectively, in the gastric wall from both injured and wound‐free areas. The healing process was more rapid in ranitidine‐treated animals than in controls. After four weeks, however, the lesion in nine out of ten animals had healed in the ranitidine‐treated group and seven of nine rats in the control group. At that time the amounts of hydroxyproline, DNA and RNA did not differ between the two groups. These findings may be taken as an indication that the tissue components of the healed lesions were similar in ranitidine‐treated rats and in the saline controls, i.e. the different speeds of the healing process did not seem to influence the components of the scar tissue.
The pathology of Shigella flexneri infection in Rhesus monkeys: An endoscopic and histopathological study of colonic lesions
Authors: Katakura SHIGEHIRO , Reinholt FINN P. , KÄRnell ANDERS , Huan PHAM THI , Trach DANG DUC , Lindberg ALF A.
Published: August 2009

Twenty‐two Rhesus monkeys were orally fed 1 times 1011 live virulent Shigella flexneri of either serotypes lb, 2a, 4a or Y. On the basis of colonoscopic findings they were classified into: group A – normal endoscopic picture (10 monkeys), and group B – pathological endoscopic picture (12 monkeys). Pathological findings, distributed over the entire colon, were seen as either red patches (± erosions) or diffuse lesions, i.e. fragile red mucosa, mucosal bleeding and broad edemas. Histopathological examination of concomitant biopsies showed an acute inflammation restricted to the mucosa in 8/12 of group B as compared to 2/10 of group A. The Shigellae were most commonly demonstrated in the surface epithelium and more rarely in the deep layer of the lamina propria. Immunohistochemical staining, using monoclonal antibodies directed against Shigella flexneri O‐antigenic polysaccharide, showed a high correlation with histopathological findings. Clinically all 10 monkeys in group A remained healthy, whereas 7/12 (all displaying histopathological signs of acute inflammation) in group B developed dysenteric symptoms. Colonoscopy should be combined with histopathological and immunohistochemical examinations of biopsies to study the pathological events taking place in the colon tissue during the course of a Shigella infection and will be of great value to assess the protective efficacy of S. flexneri vaccine candidates.
Stability of the nucleotide sequence of the phosphoprotein gene of measles virus during lytic infections
Authors: Kalland KARL‐H. , HÅVarstein LEIV SIGVE , Endresen CURT , Haukenes GUNNAR
Published: August 2009

Three clones with cDNA inserts encoding large portions of the measles virus phosphoprotein mRNA were characterized and compared with a previously published sequence of the Edmonston strain of measles virus. The two cloned viruses were separated by more than 100 passages. Only one out of 1477 nucleotides differed in the two sequences reflecting a very low mutation rate of the phosphoprotein gene during dilute lytic passages. The discovery that a third reading frame in the phosphoprotein gene may code for a novel peptide chain in addition to the P and C peptides may explain some of the high stability of the gene. The new reading frame was accessed by a translational shift caused by insertion of one extra G at a particular site in one of three otherwise identical cDNA sequences. A discrepancy was also found between the presumably high error rate of viral RNA polymerases and the stability of nucleotides in which mutations would not lead to amino acid substitutions. A few errors in the previously published sequence were discovered and the corrections are presented.
Detection of Cytomegalovirus‐infected cells in autopsy material by in situ hybridization
Authors: Andersen CLAUS B.
Published: August 2009

A non‐radioactive in situ hybridization (ISH) method was elaborated to detect cytomegalovirus (CMV)‐infected cells in tissue specimens processed for diagnostic routine histopathology. A biotinylated CMV‐DNA probe was hybridized following a) four different enzymatic predigestions, b) progressively increasing denaturation periods, and then detected by c) streptavidin‐biotin, d) a monoclonal antibody against biotin using a three‐stage alkaline phosphatase anti‐alkaline phosphatase (APAAP)‐technique, and e) combining c + d. Autopsy specimens obtained from an infant with acquired CMV‐infection, six patients with AIDS, five patients clinically and serologically without CMV‐infection, and preoperative needle core biopsies from six renal allografts served as material. ISH was specific and more sensitive when compared to immunohistochemical (IMH) detection of CMV‐antigens by a monoclonal antibody. ISH was concluded to be a rapid, practical, and sensitive tool in daily diagnostic histopathology.
Bromodeoxyuridine labeling studies on the proliferation of intestinal mucosal mast cells in normal and athymic rats
Authors: Arizono NAOKI , Shiota TSUNEZO , Yamada MINORU , Matsumoto YOSHITSUGU , Yoshikawa HISAO , Matsuda SHINJI , Tegoshi TATSUYA
Published: August 2009

The proliferation of mucosal‐type mast cells (MMC) in rat small intestine was studied using a bromodeoxyuridine (BrdU)‐labeling method. After 24‐h cumulative injections of BrdU into adult SD rats, 3.5% of MMC were labeled, while only 0.3% and 0.1% of mast cells were labeled in back skin and ear respectively. From the results, it was concluded that MMC division occurred more than 10 times as frequently as the division of skin mast cells. Similar results were obtained in athymic adult rats (F344/N Jcl‐rnu) in which the number of MMC was similar to that in heterologous animals. Thus, thymic factor(s) or T cells may not have an important role in MMC division in normal states. When SD rats were infected with Nippostrongylus brasiliensis, vigorous proliferation of MMC was brought about 13 to 15 days after infection. At that period, 40% of MMC were labeled by a single injection of BrdU, and 85% of MMC were labeled by 9‐h cumulative injections of BrdU, with the result that most MMC rapidly proliferated in the intestinal mucosa during this period. Mitotic figures of MMC were sometimes observed. On the contrary, hyperplasia of MMC was not observed in athymic rats infected with nematodes. Therefore, MMC hyperplasia after nematode infection is dependent on thymic factor or T cells, and its mechanism is different from that of MMC division in normal states, in which thymic factor(s) or T cells are not essential.
Histopathological features in stage I non‐seminomatous testicular germ cell tumours correlated to relapse
Authors: Jacobsen G. KRAG , RØRth M. , ØSterlind K. , VON DER Maase H. , Jacobsen A. , Madsen E. L. , Pedersen M. , Schultz H. ,   THE DANISH TESTICULAR CANCER STUDY GROUP
Published: August 2009

156 patients with stage I non‐seminomatous testicular germ cell tumour entered a countrywide randomized study comparing the effect of postoperative irradiation of retroperitoneal lymph nodes with surveillance only. A total of 150 patients were included in this investigation of factors associated with increased risk of relapse. Thirty‐four patients (23 percent) had a relapse, i.e. 11/67 (16 per cent) in the former and 23/83 (28 per cent) in the latter group. Histopathological studies of the orchidectomy specimens were performed in order to identify features associated with relapse. Pure embryonal carcinoma and vascular involvement were significantly associated with increased risk of relapse. Thus, the relapse rate in the observation group in patients with pure embryonal carcinomas was 50 per cent (5/10) and 36 per cent (19/53) in all patients with tumours with vascular involvement.
Heat‐shock proteins in autoimmune arthritis: A critical contribution based on the adjuvant arthritis model
Authors: VAN Eden WILLEM
Published: August 2009

Recognition of self protein epitopes, apart from those engaged in idiotypic network interactions and MHC restriction, is probably a physiological event in the normal functioning immune system. Furthermore T and B cells recognizing self antigens can be easily cloned from healthy individuals and sometimes be shown to confer autoimmune disease by passive transfer in the experimental situation. The issue is how potentially autoaggressive cells can become activated and how such activity can be contained safely. Experimentally, autoimmune disease can be evoked by immunization with autoantigens (encephalomyelitis, thyroiditis etc.) or with foreign antigens that feature antigenic relationships with self antigens (adjuvant arthritis). In both situations transfer of disease has been shown with cloned T cells of a single specificity. In addition, specific control of disease using the same cloned T cells has been achieved. Adjuvant arthritis has been illustrative in these respects. By means of specificity analysis of cloned T cells, a 65 kD heat shock protein of mycobacteria was identified as crucial in the disease. Immunization with this antigen has been found to prevent the development of disease, including forms elicited without mycobacterial involvement. Furthermore, vigorous immunological responses to HSP65 were found both in experimental animals and also in humans as a consequence of exposition to various infectious organisms. By their conserved nature HSPs have ample potential for dangerous mimicry. Recent evidence accumulated suggesting that the same HPS65 may be crucial in human chronic arthritis as well. Therefore it is hoped that extrapolation of the experimental findings to the human situation will help the development of specific means, either T cells or antigens, to control spontaneous autoimmune arthritis in man.
Immunosuppression in murine malaria: Suppressor role of macrophages and their products during acute and chronic Plasmodium berghei infection
Authors: Wangoo A. , Ganguly N. K. , Mahajan R. C.
Published: August 2009

Marked suppression of IgM, IgG and IgA plaque forming cells was observed in mice immunized with sheep erythocytes (T‐dependent antigen) during acute Plasmodium berghei infection whereas during chronic infection mild immunosuppression was observed. When mice were immunized with polyvinylpyrrolidone (T‐independent antigen), suppression in the number of plaque forming cells was observed at higher parasitaemias only during acute infection whereas during chronic infection the number remained within normal limits. Adoptive transfer studies suggested that the immunosuppression was mainly mediated by macrophages and their products. The suppressor role of macrophages and their products during acute malarial infection is highlighted.
Different cell surface and phagocytic properties in mononuclear phagocytes from blood and alveoli
Authors: SkÖLd C. M. , Eklund A. , HalldÉN G. , Hed J.
Published: August 2009

Flow cytofluorometry was used to compare blood monocytes (BMs) and alveolar macrophages (AMs) from the same nonsmoking subject (n = 13). Autofluorescence was quantified, cell surface markers (HLA‐DR, CR3) were detected by monoclonal antibodies, and phagocytic ability was determined using C3b‐coated yeast particles. AMs expressed more HLA‐DR (p < 0.001) and CR3 (p < 0.01) on their surfaces than did BMs. The phagocytic capacity was enhanced in AMs compared to BMs (p < 0.001) and the cells showed an increased autofluorescence (p < 0.001) in the alveoli compared to blood. The findings suggest that the mononuclear phagocyte is activated when it migrates from blood to alveoli in order to adapt to the milieu in the alveolar space.
Typing of polyagglutinable Pseudomonas aeruginosa isolates from cystic fibrosis patients
Authors: Ojeniyi B. , Wolz C. , DÖRing G. , Lam J. S. , Rosdahl V. T. , HØIby N.
Published: August 2009

The study assesses the reproducibility, typability and discriminatory power of several typing methods for Pseudomonas aeruginosa isolated from cystic fibrosis patients. 178 polyagglutinable Pseudomonas aeruginosa isolates from cystic fibrosis patients were serotyped using polyclonal sera and monoclonal antibodies, phage typed, pyocin typed and reverse phage typed. 31 of these polyagglutinable isolates, six monoagglutinable isolates and three nontypable isolates were also typed by means of hybridization using a DNA probe. In a comparison of the methods used, on polyagglutinable isolates only, typability was 0% with polyclonal sera, 90% with monoclonal sera, 94% with phage typing, 85% with pyocin typing, 36% with reverse phage typing and 100% with DNA‐prope typing. Using monoclonal antibodies, the reproducibility was 75%, while that of phage typing was 88%, pyocin typing 53% and reverse phage typing 62%. Typing with the DNA probe was not repeated. Using polyclonal sera, repeated typing showed that 94% of the isolates were polyagglutinable. Using phage typing, 40% of the isolates belonged to phage type 31+, while 60% were distributed amongst 32 phage types. Using monoclonal antibodies, 71% of the isolates belonged to 0‐group 3, and these isolates showed 16 different phage types. Subdisivion of the phage types was further achieved by both pyocin typing and reverse phage typing. The DNA probe typing made it possible in some cases to discriminate between isolates which were otherwise found identical with the conventional typing methods, while in other cases typing with the DNA probe recorded as identical isolates which conventional methods had typed as being different. These differences may be due to a high mutation rate caused by the selection pressure of antibiotics, and by the host immune response. According to our results, investigations of reproducibility and typability of old and new typing methods are essential when they are used in clinical situations. The low reproducibility of some of the typing methods in the present study affects the reliability of epidemiological investigations in cystic fibrosis patients. Usage of only one method may not be sufficient in cases of polyagglutinable strains from cystic fibrosis patients.
Distribution in tissue sections of the human groEL stress‐protein homologue
Authors: Evans D. J. , Norton P. , Ivanyi J.
Published: August 2009

A monoclonal antibody (ML30) raised against the 65 kDa heat‐shock protein of mycobacteria showed widespread staining of sections from standard paraffin‐embedded human tissues. The staining had a granular pattern and was particularly marked in cells with abundant mitochondria. Increased staining was observed in the synovial lining, histocytes and in the endothelium of reactive and rheumatoid synovium; it was also increased in the reactive lung alveolar lining. It is suggested that the antibody identifies an epitope in mitochondria of a protein homologous with the groEL heat‐shock protein of bacteria.
A comparison of different challenge methods for induction of atrophic rhinitis in pigs
Authors: Chung WEN‐BIN , BÄCkstrÖM LENNART R. , Conrad THERESA , Collins MICHAEL T.
Published: August 2009

Transmission and development of atrophic rhinitis (AR) was studied in 5‐ to 15‐week‐old pigs (Groups 2–7) originating from a herd free of AR, and compared to unexposed healthy pigs (Group 1), and pigs from a herd with endemic AR (Group 8). At the start of the trial, pigs in Groups 2–5 were challenged intranasally twice a week for 3 weeks with pure cultures of bacteria originating from the endemic AR herd: NontoxigenicPasteurella multocidatype A (PmA) plusBordetella bronchisepticaphase 1 (Bb) (Group 2);PmA+ toxigenicPmtype D (PmD) (Group 3);PmDonly (Group 4); andPmD + Bb(Group 5). Group 6 pigs were challenged with nasal wash of pigs from the endemic AR herd, and Group 7 pigs were challenged by being housed together in the same pen with Group 8 pigs throughout the study. Nasal swabs of all pigs were cultured 5 times during the study. Serum was collected at 6 weeks post challenge. Average daily gain (ADG) and turbinate lesions (turbinate gross lesions by visual scoring and by Turbinate Perimeter Ratio, TPR, scoring, and histopathological lesions) were measured at the time of slaughter at 15 weeks of age. Mean TPR value for the Group 1 pigs was 1.64, which was significantly (P < 0.05) different from the mean TPR value of 0.58 for the pigs from the endemic AR herd (Group 8), the 0.79 value for Group 6 pigs, and 1.03 value for Group 7 pigs. Of pigs challenged with pure bacterial cultures, only Group 5 (PmD + Bb) developed significant AR (mean TPR = 1.24). Only one pig in each of Groups 2 and 3, and two pigs in Group 4 showed TPR values indicative of AR (TPR < 1.30). However, histopathological examination showed that those pigs were recovering from the infection 7 weeks post challenge. Constant exposure to certain bacteria or other factors in nasal washings, stress of crowding or poor environmental conditions might be required to experimentally produce AR in 5‐week and older pigs similar to that in naturally infected pigs. There was no relationship betwen turbinate lesions and the isolation frequency or quantity ofPmA, PmD, orBb.Antibody levels againstPmAorPmDhad moderate to high correlation with TPR values (r = ‐0.694 and ‐0.503 respectively). ELISA values also corresponded well with the type of bacteria inoculated in each group of pigs and appeared to be a sensitive test forPmA, PmD, andBbinfections in pigs. Pigs with TPR values less than 1.30 grew less (ADG = 588 g) than pigs with TPR values greater than 1.30 (ADG = 661 g), but the difference was not significant (P = 0.083). Pneumonic lesions were also present in some of the pigs with low ADG.
Adherence ofBordetella bronchisepticaandPasteurella multocidato swine nasal ciliated epithelial cellsin vitro
Authors: Chung WEN‐BIN , Collins MICHAEL T. , BÄCkstrÖM LENNART R.
Published: August 2009

A bacterial adherence assay using swine nasal turbinate fragments was established. Turbinate fragments were incubated withBordetella bronchisepticaorPasteurella multocidatype D at different concentrations or for different inbucation times at 37°C on a shaker at 120 rev/min.B. bronchisepticaphase I strains exhibited strong adherence to swine nasal ciliated epithelial cells. The number of adherent bacteria per cell increased when the bacterial concentration or incubation time increased (0, 15, 30, and 60 min); however, the number of adherent bacteria decreased after 3 or 6 hours' incubation due to the loss of cilia from cells. The optimal bacterial concentration and incubation time were 1 times 109organisms/ml and one hour respectively, which resulted in 7.48 ± 0.66 (Mean ± SEM;B. bronchisepticastrain 03) and 9.31 ± 0.54 (B. bronchisepticastrain 013) adherent bacteria per cell. In contrast toB. bronchisepticaphase I strains, rough phase strains ofB. bronchisepticaand allP. multocidastrains tested showed no adherence to swine nasal ciliated epithelial cells. AllB. bronchisepticaphase I strains could agglutinate calf RBC but rough phase strains could not. Furthermore, pretreatment ofB. bronchisepticaphase I organisms with 1 mg/ml or 2 mg/ml of trypsin significantly inhibited the adherence ofB. bronchisepticato ciliated epithelial cells; however, trypsin (2 mg/ml) treatment of bacteria did not decrease their ability to agglutinate calf RBC. From these results we conclude that, in addition to hemagglutinin, other proteinaceous components exist on the surface of virulentB. bronchisepticathat are sensitive to 2 mg/ml trypsin; these are suggested to be the adhesins for the adherence ofB. bronchisepticato swine nasal ciliatd epithelial cells.
Antibiotic susceptibility and β‐lactamase production in clinical isolates of Enterobacter spp.
Authors: Lindh ELISABET , Dornbusch KATHRINE , Jalakas KIRSTI , Forsgren ARNE
Published: August 2009

The in vitro susceptibility of 237 clinical isolates of Enterobacter spp. (E. aerogenes, E. agglomerans and E. cloacae; 41, 64 and 132 respectively) to 16 different antibiotics is described. Four quinolones (ciprofloxacin, lomefloxacin, norfloxacin and ofloxacin), two new cephalosporins (cefpirome and cefepime) and imipenem, all showed high activity against the three Enterobacter species tested (MIC50≤ 0.125 mg/l, MIC90≤ 0.5 mg/l). Also the aminoglycosides gentamicin and tobramycin were highly active antibiotics (MIC50≤ 0.5 mg/l, MIC90≤ 1.0 mg/l). The susceptibility of β‐lactam‐antibiotics to β‐lactamase produced by Enterobacter spp. was evaluated, and imipenem and cefepime were found to be most stable. Different methods for detection of inducible β‐lactamases were used, the agar dilution method being more sensitive than the double‐disc diffusion test. Elevated β‐lactamase production was detected, via induction, in 83% of E. aerogenes strains and 70% of E. cloacae strains, with cefamandole used as the substrate and cefoxitin as the inducer. Constitutive, high level enzyme production was detected in 7 and 13% respectively of the E. aerogenes and the E. cloacae strains. In all the strains of E. agglomerans, 10% of E. aerogenes and 13% of E. cloacae, no β‐lactamases could be detected with the methods studied.
Attachment of staphylococci to silicone catheters in vitro
Authors: Espersen FRANK , Wilkinson BRIAN J. , Gahrn‐Hansen BENTE , Rosdahl VIBEKE THAMDRUP , Clemmensen INGE
Published: August 2009

The adherence of radiolabeled staphylococci to silicone catheters was investigated in vitro. Staphylococcus aureus and Staphylococcus epidermidis strains bound to the same extent to the catheters. Also, S. epidermidis strains isolated from patients with plastic‐related infections showed binding similar to that of other S. epidermidis strains. By preincubation of catheters the influence of purified staphylococcal cell surface components on the binding was evaluated. The most potent inhibitors of the binding of S. aureus were the two surface proteins, clumping factor and protein A, and the cytoplasmic membrane. Surface proteins and the cell membrane of S. epidermidis also blocked the binding. Only protein‐containing surface proteins inhibited the binding. The production of slime correlated with the degree of S. epidermidis binding. Human plasma and serum, as well as purified albumin and IgG, inhibited the binding of both staphylococcal species. Fibrinogen, and to a certain extent fibronectin, inhibited the binding of S. epidermidis, while both these purified plasma proteins enhanced the binding of S. aureus.
The value of immunohistochemistry and in situ hybridization in detecting cytomegalovirus in bone marrow transplant recipients
Authors: Rasing L. A. J. , DE Weger R. A. , Verdonck L. F. , VAN DER Bij W. , Compier‐Spies PH. I. , DE Gast G. C. , VAN Basten C. D. H. , Schuurman H. J.
Published: August 2009

Autopsy tissues of 19 patients with complications after bone marrow transplantation (BMT) were analysed for the presence of cytomegalovirus (CMV) using histochemical methods. CMV antigens were detected by antibodies to CMV Immediate Early Antigen (IEA) or CMV Late Antigen (LA). CMV‐DNA was detected by DNA in situ hybridization (DISH). IEA was detected in one or more tissues in 79% of 14 patients from whom frozen tissue was available. CMV‐DNA was detected on paraffin sections in 84% of all 19 patients. CMV components were present in all organs studied; the highest incidence was found in lung, gastrointestinal tract and kidney. In histology, only 37% of patients showed signs of CMV infection by the presence of cytomegalic cells with nuclear inclusions (or so called “owl eye cells”). In tissue culture, only 33% of 15 patients were CMV positive. Serologically, 68% of all patients had active CMV infection, as indicated by a rise in antibody titres. We conclude that the quick detection of CMV IEA and CMV‐DNA has a high sensitivity and predictive value, which is comparable to or exceeds the serological detection of CMV.
Clinically undiagnosed malignant tumours found at autopsy
Authors: Karwinski B. , Svendsen E. , Hartveit F.
Published: August 2009

In a series of over 20,000 autopsies carried out over a 25‐year period, 700 cancers (11%) were found in patients in whom the diagnosis of cancer had not been considered relevant clinically. In over half of them the unrecognized tumour was considered an incidential finding and thus of no importance to the individual concerned, though of epidemiological concern. As expected from the literature the main organs involved were kidney and prostate, with stomach cancer in third place. In contrast, the unrecognized cancers that caused death were most often from the pancreas or lung, again with the stomach in third place. These patients with stomach or lung cancer were frequently thought to have died of other diseases in the gastrointestinal/respiratory tract, and the disease cited was often present though not the cause of death. These patients tended to be older than those with clinically recognized disease, and had been hospitalized late in the course of their fatal illness, at which time clinical recognition would not have influenced the outcome. Earlier recognition of this type of cancer is thus essential. This cannot be achieved without definition and further analysis of the entities concerned.
Serogroup determination of Neisseria meningitidis by whole‐cell ELISA, dot‐blotting and agglutination
Authors: Rosenqvist EINAR , Wedege ELISABETH , HØIby E. ARNE , FrØHolm L. ODDVAR
Published: August 2009

Two new methods for serogrouping of meningococci, whole‐cell ELISA and dot‐blotting, with monoclonal antibodies against serogroups A, B, C, Y and W135 were compared with slide‐agglutination applying polyclonal sera. In addition to a panel of strains with previously determined serogroups by slide‐agglutination, two strain collections of meningococci were studied: 1) 50 strains isolated from patients with systemic meningococcal disease in Norway during the winter 1987–1988; 2) 133 throat strains isolated from asymptomatic carriers over the same period. For the disease strains all three methods gave identical results, whereas some carrier strains which were non‐agglutinable or polyagglutinable by slide‐agglutination were serogroupable by the two other methods. All the systemic strains and about half of the carrier strains were serogroupable. We find that whole‐cell ELISA and dot‐blotting are specific, easy to read and more sensitive compared to slide‐agglutination, but the former methods are at present limited by the availability of monoclonal antibodies against only serogroups A, B, C, Y and W135.
Splenic autotransplantation in nude mice
Authors: Sasaki KATSUNORI
Published: August 2009

Splenic fragments were autotransplanted into the greater omentum of nude mice (nu/nu) and normal mice to investigate the formation of white pulp in splenic regeneration. The formation of white pulp began from day 5 after transplantation in both groups. Two patterns were found. In one pattern, follicles containing only B cells were first formed and developed into white pulp with poorly developed periarterial lymphoid sheaths (PALS). Most of the white pulp in nude mice and about one third of white pulp formed at day 28 in normal mice followed this pattern. In the second pattern, PALS were first formed and afterwards follicles appeared in the periphery of PALS. This was mainly seen in normal mice. Thus splenic autografts in normal mice contain white pulp with poorly developed PALS, and the homing of T cells begins later than that of B cells. This suggests the possibility of incomplete cellular immunity.
Neisseria meningitidis group B capsular polysaccharide
Authors: Hoddevik GUNNAR MARTIN , Baastad KIRSTEN LYCHE , Bukholm GEIR , Bryn KLAUS , BØVre KJELL
Published: August 2009

A total of 41 Neissera meningitidis isolates were analyzed for capsular polysaccharide (CP) and lipopolysaccharide (LPS) release and filtrability. Twenty‐two of these isolates were serogroup B from blood or cerebrospinal fluid of patients, five were group B isolates from healthy throat carriers, and 14 were nongroupable (acapsular) meningococcal isolates from healthy carriers. Filtration of liquid whole‐cell cultures through cellulose acetate‐nitrate filters resulted in distinctly lower LPS filtrate activity for acapsular than for capsular meningococci (p < 0.001). On the other hand, when polysulfone membrane filtration was performed, filtrates from acapsular and capsular meningococci contained LPS in similar amounts. These results indicate that LPS‐containing particles released from acapsular isolates are larger or more aggregated than corresponding CP‐ and LPS‐containing particles released from capsular isolates. The LPS released from acapsular isolates apparently are more efficiently retained by adsorption to cellulose acetate‐nitrate. From the capsular isolates comparatively more CP than LPS appeared to be released, as related to cell‐bound amounts. The total amounts of CP and released, filtrable LPS through cellulose acetate‐nitrate filters were both relatively low and had similar values for capsular carrier meningococci and systemic isolates from mild meningococcal disease. The amount of CP released and passing this type of membrane was significantly higher for systemic isolates from severe septicemia than from mild meningococcal disease (p = 0.03).
Quantitation and biological activities of native tumour necrosis factor from LPS‐stimulated human monocytes
Authors: Fomsgaard A. , Freudenberg M. A. , Bendtzen K. , Galanos C.
Published: August 2009

Human mononuclear cells (MNC) were stimulated in culture with LPS to produce tumour necrosis factor (TNF). The natural tumour necrosis factor (nTNF) was quantitated by ELISA and immunoblotting using rabbit antibodies to human recombinant TNF (rTNF) and the biotin‐avidin‐peroxidase system. Biologically active nTNF was determined by its cytotoxic activity for actinomycin‐D treated mouse fibroblasts and by its lethal effect in D‐galactosamine sensitized endotoxin‐resistant mice. Two different LPS preparations (Salmonella abortus equi and Pseudomonas aeruginosa) induced the formation of comparable amounts of nTNF. MNC from different donors, however, showed large variations in their ability to produce nTNF. The amount of nTNF induced in response to LPS could be enhanced by priming the MNC with interferon. The amounts of nTNF determined by ELISA generally correlated well with the activity of the nTNF in the two biological assays. On a weight basis, the lethal activity of nTNF in D‐galactosamine treated mice was very similar to that of human rTNF. Immunoblotting revealed a single band of nTNF with the same molecular weight (17 kD) as human rTNF. The lethality induced by nTNF was inhibited by rabbit anti‐human rTNF antibodies.
Studies on the polyisoprenoid composition in hepatocellular carcinomas and its correlation with their differentiation
Authors: Eggens I. , Elmberger P. G.
Published: August 2009

The levels of cholesterol, ubiquinone and dolichol and the polyprenol composition of dolichol in human hepatocellular carcinomas (hepatomas) with different degrees of differentiation were analyzed and compared with healthy liver tissue. Dolichols were also analyzed in liver metastases. The total level of cholesterol was increased, while the levels of dolichol and ubiquinone were decreased in all hepatomas, but no correlation between these levels and the degree of differentiation of the hepatomas could be observed. The level of dolichol decreased more in the hepatomas than in the liver metastases. The dolichol fraction from hepatomas with a low degree of differentiation contained higher relative amounts of short polyisoprenols (D17) and slightly lower relative amounts of D21 compared with healthy liver tissue, metastatic liver tumors or hepatomas with a high degree of differentiation. The significance of the lipid values found in the different groups is discussed.
A subdivision of strains of Staphylococcus aureus in the 94,96 complex by means of experimental phages
Authors: Rosendal KIRSTEN , Bentzon MICHAEL WEIS , DE Saxe MAUREEN , Rosdahl VIBEKE THAMDRUP
Published: August 2009

In order to facilitate epidemiological investigations a subdivision of Staphylococcus aureus strains belonging to the 94,96 complex by means of two experimental phages, 16 and 47A, was performed. These phages were selected from the nine experimental phages initially examined because they gave the greatest discrimination. On the basis of reactions with these two phages, 2199 isolates which reacted with phages 94 and 96, and 773 isolates which reacted with phage 96 alone, were each subdivided into two major and two minor groups. Strains with different phage patterns were in a few cases (2/64) isolated from the same deep body site in a patient, and lysogenisation experiments suggested that differences in phage patterns were determined by the presence of prophages. Strains with the phage patterns 94/96 and 96 were found to be unevenly distributed throughout Denmark. This regional distribution suggested that particular strains might predominate in some areas. The extended phage patterns with the experimental phages did not give any retrospectively useful epidemiological information. It is proposed that in future phages 16 and 47A be used for specific investigations into the sources and relatedness of strains involved in small incidents.
A qualitative and quantitative method for in situ characterization of the inflammatory response in experimental myocarditis
Authors: Fohlman J. , Friman G. , IlbÄCk N. G. , ÅKesson Å. , Huber S.
Published: August 2009

Coxsackie virus B3 causes myocarditis in Balb/c inbred mice. In this model mortality is about 60% on day 7 and 95% on day 12 after inoculation. Significant inflammatory infiltrates appear on day 7. Recent research has focused on the immune response to explain the ensuing tissue damage. Thus we were interested in mapping cellular interaction and time course to understand the development of a possible autoimmune attack. By a newly developed immunohistochemical staining technique single lymphocytes could be visualized and different lymphocyte subpopulations enumerated. Stained cells were easily detectable and readily distinguishable from non‐stained cells. Before day 7 Thcells could only occassionally be seen. The number of pan T lymphocytes increased almost 10‐fold from day 7 to 12. The T helper/T killer ratio did not change significantly but indicated a predominant increase in Tk‐cells. The B cell population also increased, roughly about ten times. The number of class II positive cells was constant. No expression of Il‐2 receptor was found. In preliminary studies, exercise and methylprednisolone treatment tended to influence the expression of class II antigens. Cyclophosphamide and cyclosporine A reduced the number of inflammatory cells more than 10‐fold. No clear concurrent reduction in mortality was observed.
The influence of barley fibre on bile composition, gallstone formation, serum cholesterol and intestinal morphology in hamsters
Authors: Zhang JIE‐XIAN , Bergman FRANK , Hallmans GÖRAN , Johansson GERD , Lundin EVA , Stenling ROGER , Theander OLOF , Westerlund ERIC
Published: August 2009

Frequency of gallstones, concentration of bile acids and cholesterol in bile, concentration of cholesterol in serum, and structure of the small intestinal mucosa were analyzed in male Syrian Golden hamsters fed a stone provoking fibre‐free diet with or without supplementation of brewer's spent grain (BSG), a concentrated barley fibre source from the by‐product of brewing. A significantly lower frequency of gallstones was found in the animals with 10% BSG dietary supplementation. Addition of 30% BSG after an initial 6‐week period with a fibre‐free, stone provoking diet seemed to dissolve previously formed gallstones. Total bile acid concentration was higher in bile from animals given a diet supplemented with 10% BSG. In addition, the cholesterol concentration in both serum and bile was lower in the 30% BSG supplemented group. Structurally, a 10% BSG supplementation decreased ileal epithelium height whereas a high supplementation (30%) of BSG induced a decrease in epithelial height both of jejunal and ileal mucosa. The results show that BSG has significant effects on the metabolism of bile acids and cholesterol as well as on the morphology of the small intestinal mucosa.
Genes for MHC, TCR and MIS determine susceptibility to collagen induced arthritis
Authors: David CHELLA S.
Published: August 2009

Type II collagen induced arthritis (CIA) in mice is an animal model for polyarthritis. The susceptibility to the disease is linked to the class II genes of H‐2 gene complex (MHC). The susceptibility requires T cells expressing certain receptors coded by the Vβ genes. Further, the MIs gene products in combination with the class II molecules can up‐or down‐regulate the T cells involved in the disease. The disease is mediated by the presentation of an arthritogenic epitope on the collagen type II peptide by the MHC class II molecule, which is recognized by a T cell expressing certain Vβ receptors, triggering the autoimmune response. These studies point out possible mechanisms of rheumatoid arthritis in humans and suggest various methods of immune intervention to down‐regulate the disease.
Cell‐mediated immune responses to soluble Plasmodium falciparum antigens in residents from an area of unstable malaria transmission in the Sudan
Authors: Hviid LARS , Theander THOR G. , Jakobsen PALLE H. , Abu‐Zeid YOUSIF A. , Abdulhadi NASRELDIN H. , Saeed BAKRI O. , Jepsen SØREN , Bayoumi RIAD A. L. , Bendtzen KLAUS , Jensen JAMES B.
Published: August 2009

This paper describes immune responses to P. falciparum infection in individuals living in an area of highly seasonal, unstable malaria transmission. The in vitro cellular immune responses of peripheral blood mononuclear cells (PBMC) from 36 Sudanese donors to a complex of affinity purified soluble P. falciparum antigens (SPag) and two components thereof (Ag1 and Ag7) were examined and compared to humoral immune parameters. In 29/36 Sudanese donors, SPag induced a significant lymphoproliferative response in vitro. In contrast only 3/27 Danish donors never exposed to malaria responded to SPag. Ag1 and Ag7 induced significant lymphoproliferation in 9/34 and 13/36 Sudanese donors respectively, whereas no Danish donors responded. Significant interferon‐gamma production was observed in 16/27, 1/5 and 3/12 Sudanese donors when stimulated by SPag, Ag1 and Ag7 respectively. Lymphoproliferative responses to SPag correlated with proliferative responses to Ag1 and Ag7, and with Spag‐induced interferon‐gamma production. These results indicate that T‐cell clones recognizing epitopes on Ag1 and Ag7 have been expanded in the studied population as a result of exposure to P. falciparum infection. The T‐cell parameters did not correlate with the presence of antibodies to SPag, Pf155/RESA or a crude parasite sonicate or with the schizont IFA titers of the plasma. This indicates that parameters outside the degree of exposure to P. falciparum influence the cellular immune responses to malaria.
Proliferation of adherent synovial fluid cell cultures is modified by eicosanoids
Authors: Andreasen A. , Hansen E. S. , Fogh K. , Kragballe K. , Herlin T.
Published: August 2009

Synovial fluid cells obtained from a carragheenan‐induced chronic arthritis in the juvenile dog knee were allowed to adhere and proliferate in culture flasks. After twelve days secondary cultures were made and either 10−8M leukotriene B4 (LTB4), 25 μM 15‐hydroxy‐eicosatetraenoic acid (15‐HETE), or both, were added and the cells were cultured for another 6 days. LTB4 is generated via the 5‐lipoxygenase pathway of arachidonic acid metabolism and stimulates a number of phagocyte functions. Compared to control cells LTB4 increased proliferation in 9 out of 10 cell cultures (p > 0.05). The 15‐lipoxygenase product, 15‐HETE, is not proinflammatory and is an endogenous inhibitor of 5‐lipoxygenase. Addition of 15‐HETE decreased proliferation of cell cultures by 23% (p > 0.01). It is speculated that LTB4 in addition to its effect on phagocytes may play a role in synovial hyperproliferation observed in arthritis.
Influence of whole human milk, and fractions thereof, on inclusion‐formation of Chlamydia trachomatis in McCoy cells
Authors: Elbagir ADIL , Petterson MARGARETA , Lindahl MATS , Genc MEHMET , FrÖMan GUNNAR , MÅRdh PER‐ANDERS
Published: August 2009

A study of the effect of human breast milk, and components thereof, on the capacity of Chlamydia trachomatis to form inclusions in cycloheximide‐treated McCoy cells, was undertaken. Pooled whole milk collected during the first week of breast feeding caused a concentration‐dependent inhibition of the chlamydial inclusion‐formation. The activity resided in the fat and fat globule membrane (FGM) components of the milk. The active principle in the FGM fraction is heat‐stable and pronase‐sensitive, but resistant to both neuraminidase and periodate. Immunoglobulins was not responsible for the inhibition. Whey and casein fractions of milk increased the chlamydial inclusion‐formation. The activity of the whey was heat‐stable, dose‐related, and had a mol.wt. of 12,000. The casein fraction was still active after heat treatment. Whey samples collected up to 28 days after delivery varied slightly in their stimulatory activity, with an optimum between the 7th and 14th days. The present study demonstrated a multieffect of breast milk on chlamydial inclusion‐formation: an inhibitory activity due to a protein compound as well as another factor in the fat fraction and an enhancing effect due to a heat‐stable factor(s).
Fibronectin: A discrimination marker between small invasive carcinomas and benign proliferative lesions of the breast
Authors: Christensen LISE
Published: August 2009

The connective tissue glycoprotein, fibronectin (FN) is known to be increased in the stromal desmoplastic response associated with invasive breast carcinoma (IBC). In a previous study of 77 IBCs we found that all tumours, irrespective of desmoplasia, displayed an intense stromal immunoreactivity for FN, although those with a high metastatic potential tended to lack this in foci along their infiltrating border (Chrislensen et al. 1988). In order to test the diagnostic value of this observation, FN immunoreactivity was estimated semiquantitatively in 131 small primary IBCs, 23 in situ carcinomas, 20 borderline lesions, 142 benign proliferative breast lesions and 35 samples of normal breast tissue. All IBCs, including those without apparent desmoplasia, were accompanied by an intense stromal staining for FN forming a diffuse or pericellular pattern around the tumour cells (FN positive reaction). This was not seen in the benign lesions (FN negative reaction) with the exception of an FN positive, linear staining around some cysts and ectatic ducts and a diffuse staining of the connective tissue core of some intraductal papillomas. Intraductal carcinomas, multiple papillomas and radial scars could display a substantial staining for FN, sometimes enclosing small islands or tubules of epithelial cells. However, the fact that all IBCs of this study produced an overall reliable and consistent FN positive stromal reaction, which was only interrupted in minute foci along the infiltrating border in a few of the largest tumours (11/131), makes immunohistochemical staining for FN an excellent adjunct to morphology in tracing small, primary IBCs.