bluuebunny/crossref_metadata_2025 huggingface dataset:
Anti‐CD4 monoclonal antibody treatment in combination with total lymphoid irradiation and cyclosporin A in hamster‐to‐rat cardiac transplantation
Authors: STEINBRÜCHEL DANIEL A. , NIELSEN BJARNE , KEMP EJVIND
Published: August 2009

Significantly prolonged graft survival (GS) of hamster hearts transplanted heterotopically into rats can be achieved by different immunosuppressive treatment strategies. The exact mechanism of graft rejection is unclear, but it seems to be a primarily humoral, antibody‐mediated type of rejection. The histopathology of long‐term surviving grafts is controversial and the morphology of lymphoid tissue in spleens and lymph nodes as the possible site of anti‐donor antibody formation has not previously been investigated. This report demonstrates a significantly prolonged GS in hamster‐to‐rat cardiac transplantation after combined treatment with total lymphoid irradiation (TLI), cyclosporin A (CyA) and anti‐CD4 monoclonal antibodies (MAb), where long‐term GS (>100 days) could be achieved in a few animals. The histopathology of heart grafts showed predominantly chronic vascular changes with endothelial proliferation, intimal thickening and vessel obliteration. No substantial cellular reactivity in terms of mononuclear/lymphoid cell infiltration could be demonstrated in rejected grafts. Spleens and lymph nodes were characterized by a profound global reduction in lymphoid tissue after preoperative TLI. Although subsequent lymphoid regeneration was depressed due to postoperative immunosuppression, a significant increase in IgM‐positive plasma cells was observed, supporting evidence of an antibody‐mediated mechanism of graft rejection. The role of CD4+ cells is unclear, but anti‐donor antibody formation might involve T‐cell help.
Mycoplasma contamination of chlamydiae isolated from clinical specimens
Authors: MESSMER TRUDY O. , BLACK CAROLYN M. , THACKER W. LANIER
Published: August 2009

Ten Chlamydia pneumoniae strains were screened for Mycoplasma contamination using two differently designed Mycoplasma‐specific polymerase chain reactions (PCR). The primers of the Mycoplasma‐specific PCR designed by Spaepen et al. (9) cross‐reacted with all of the C. pneumoniae strains giving false‐positive results. When the 10 strains of C. pneumoniae were tested for mycoplasmas with the PCR designed by Harasawa et al. (5), only 3 were positive. Mycoplasmas were cultured from these three C. pneumoniae strains confirmning the latter PCR results. The PCR of Harasawa et al. (5) was highly specific for mycoplasmas and did not cross‐react with C. pneumoniae. These findings suggest that chlamydiae should be periodically screened for Mycoplasma contamination. Careful attention to primer design is important if PCR is chosen as the screening method.
Gastropathies in the Lundehund
Authors: KOLBJØRNSEN Ø. , PRESS C. McL. , LANDSVERK T.
Published: August 2009

The mucin profiles of the gastric mucosa in Lundehunds suffering from intestinal lymphangiectasia were examined and compared to the mucin profiles in control dogs from other breeds. A previous study performed on this material had shown that all examined Lundehunds had gastritis and about 30% had gastric carcinoma. Neutral and acid mucins were identified using the periodic acid‐Schiff (PAS) and Alcian blue (pH 2.5) periodic acid‐Schiff (AB‐PAS) methods. The acid mucins were divided into sialomucins and sulfomucins based on their reaction with high‐iron diamine Alcian blue, pH 2.5 (HID‐AB). In Lundehunds with chronic atrophic gastritis in the fundic and body regions the surface and foveolar epithelium showed a predominantly normal mucin profile although some Lundehunds had a reduced mucin content. The mucous neck cells extended from below the gastric foveolae towards the muscularis mucosae. Morphometric examination showed that the abnormal presence of mucous neck cells occupied 41% of the height of the gastric mucosa in Lundehunds compared to only 19% in the control dogs (p<0.05). Of the four Lundehunds with gastric carcinoma, two possessed neoplastic cells that contained minimal or no mucins. The amount and type of mucin in the neoplastic cells of the remaining two Lundehunds varied both between individuals and within a neoplasm. This study shows that the abnormal presence of mucous neck cells and the associated pseudopyloric metaplasia comprised the predominant changes in the gastric mucin profiles of Lundehunds.
The high‐affinity folate receptor of normal and malignant human colonic mucosa
Authors: HOLM JAN , HANSEN STEEN I. , HØIER‐MADSEN MIMI , SØNDERGAARD KNUD , BZOREK MICHAEL
Published: August 2009

The hypothesis that folate depletion is a risk factor for development of colonic neoplasia prompted us to study the presence of a putative folate receptor in human colon mucosa. Binding of 3H‐folate to normal and malignant mucosa studied by equilibrium dialysis was of a high‐affinity type (K= 1010 L/mol) and displayed apparent positive cooperativity. Radioligand dissociation was slow at pH 7.4, but rapid at pH 3.5. As compared to methotrexate, 5‐formyltetrahydrofolate was a potent inhibitor of binding. Gel filtration revealed a 25 kDa and a 100 kDa peak of folate‐binding activity. Immunoreactivity studies were performed with rabbit antibodies against human 25 kDa milk folate‐binding protein. Immunoblotting showed a single band at 65 kDa, and tissue sections exhibited immunostaining of mucosal areas. The present folate receptor with characteristics similar to those of other high‐affinity folate‐binding proteins may serve as a regulator of intracellular folate concentration in colon mucosa.
Preoperative histological and cytological diagnosis and DNA ploidy assessment of localized prostatic cancer
Authors: HÄGGMAN MICHAEL , DE LA TORRE MANUEL , NORBERG MONA , BUSCH CHRISTER
Published: August 2009

The preoperative biopsy specimens of 49 men who underwent radical prostatectomy were compared with serially step‐sectioned operative specimens, in order to evaluate diagnostic accuracy as regards the presence of cancer, its grade and its DNA ploidy. Both the FNAB and TRUS‐GCB undergraded the cancer by one WHO grade in about 35–40% of cases. In cases where both biopsy types were available, the degree of undergrading was the same. Determination of DNA ploidy by the single‐cell technique from FNAB had a sensitivity for detecting non‐diploid DNA patterns of 59% whereas flow cytometric measurement of core biopsies had a sensitivity of 44.4% as regards non‐diploid DNA, when compared with operative specimens. A comparison of ploidy in core biopsies versus ploidy in fine‐needle aspirates revealed that more non‐diploid DNA patterns were diagnosed in the fine‐needle aspirates. These aneuploid patterns were not all confirmed by flow cytometric evaluation of the operative specimens, in which, however, more aneuploid patterns were diagnosed compared with the single‐cell technique from FNAB. We conclude that the standard technique with 1–3 fine‐needle aspirates or 1–3 TRUS‐GCB has a pronounced tendency by both biopsy methods to underestimate the tumour grade. DNA ploidy analysis by FNAB and the single‐cell cytometric method reveals aneupolid cell lines not found in the flow cytometric evaluation. In order to determine whether this reflects a methodological problem or a true discrimination between the ability of the two methods to find non‐diploid cell lines further studies are needed. However, for a more correct preoperative assessment of tumour grade and DNA ploidy, more extensive sampling is required.
The first fatal case of Vibrio vulnificus infection in Denmark
Authors: BOCK TROELS , CHRISTENSEN NILS , ERIKSEN NIELS H. RIEWERTS , WINTER STEEN , RYGAARD HENRIK , JØRGENSEN FRANK
Published: August 2009

Vibrio vulnificus can cause severe infections in humans and persons with preexisting liver disorders are especially at risk. In this paper we report what is to our knowledge the first fatal case of V. vulnificus infection in Denmark. The patient was a 68‐year‐old man with a history of chronic lymphatic leukemia and hepatic cirrhosis. Physicians should be aware of the clinical manifestations of this disease and should be especially attentive to patients at risk of acquiring the infection if there has been possible exposure to V. vulnificus by contact with seawater or contaminated material such as eels.
Staphylococcal neutral phosphatase
Authors: YOUSIF YEHIA , SCHILTZ EMILE , OKADA KANAME , BATSFORD STEPHEN , VOGT ARNOLD
Published: August 2009

Staphylococcal neutral phosphatase (NPtase) was purified from two Staphylococcus aureus strains by sequential high salt extraction, ultracentrifugation and ion exchange chromatography. The enzyme showed maximum phosphatase activity at neutral pH, appeared as two bands in SDS‐PAGE (31 and 32 kDa), and the isoelectric point was > 10. No close similarity between NPtase and other known bacterial proteins in respect of their N‐terminal amino acid sequences was found. Purified NPtase bound rat and human polyclonal IgG [intact and F(ab')2 fragments], IgM, IgA, intact myeloma immunoglobulins, myeloma light chains, gamma heavy chain and, with a much lower affinity, Fc fragments. Furthermore, NPtase can bind serum albumin. Heparin, a highly negatively charged molecule, significantly inhibited NPtase binding to immunoglobulins and HSA, but did not inhibit the binding of specific antibodies to NPtase; this indicates that charge interactions are important. The newly characterized staphylococcal phosphatase with binding properties for immunoglobulin is an interesting bacterial protein that could be involved in post‐infectious sequelae.
Priming of oxidative response in human neutrophils by anti‐CD18 monoclonal antibodies
Authors: FORSLUND TONY , SUNDQVIST TOMMY
Published: August 2009

Type I collagen, the most abundant protein in the body, after acid extraction adheres to and can induce a respiratory burst from neutrophils. It has been proposed that the effects of collagen are mediated via the CD 18 subfamily of integrins. In the present study, adhesion was measured by affinity chromatography in a column containing collagen‐coated microcarriers, while oxygen metabolite production was measured with luminol‐dependent chemiluminescence. Neutrophil adherence to collagen was attenuated by anti‐CD 18 monoclonal antibodies. The respiratory burst in response to collagen was not affected by the antibodies. Incubation of neutrophils with anti‐CD 18 antibodies prior to stimulation with FMLP increased both the extra‐ and intracellular respiratory burst. Treatment with antibodies prior to PMA stimulation increased only the extracellular respiratory burst. In conclusion, the respiratory burst from neutrophils is primed by pretreatment with anti‐CD 18 monoclonal antibodies. The collagen‐stimulated respiratory burst is probably also primed, but the effect is hidden by the simultaneous attenuation of adhesion.
Prognostic value of Ki‐67 expression in 182 soft tissue sarcomas. Proliferation — a marker of metastasis?
Authors: CHOONG P.F.M. , ÅKERMAN M. , WILLÉN H. , ANDERSSON C. , GUSTAFSON P. , BALDETORP B. , FERNÖ M. , ALVEGÀRD T. , RYDHOLM A.
Published: August 2009

Soft tissue sarcomas (STS) are characterized by deregulated proliferation. Ki‐67 is a cell cycle antigen which may be elevated in proliferative states. We analysed Ki‐67 expression in fixed and embedded tissues from STS in order to examine associations between proliferation, primary tumour characteristics, and metastasis. One hundred and eighty‐two adult patients with trunk wall or extremity STS were treated at our institution between 1980 and 1992 (35 developed local recurrence and 56 developed metastases). Median follow‐up time for survivors was 6 years (1–13). We used a semiquantitative score to the assess percentage of Ki‐67‐positive cells: ≥ 10% (n = 86), >10–25% (n = 57), >25–50% (n = 30), >50–75% (n = 7), >75–100% (n = 2). Increasing Ki‐67 expression correlated positively with tumour size, malignancy grade, necrosis, vascular invasion, S‐phase fraction, and metastasis. A Ki‐67 index Ki‐D 10% (n = 86) and >10% (n = 96) defined two groups who had 84% and 56% 3‐year metastasis‐free survival (p= 0.0001), respectively. Tumours with Ki‐D>10 were typically large, high grade, necrotic, DNA aneuploid, and had intravascular invasion and a higher S‐phase fraction. Ki‐67 expression may be helpful in predicting survival of patients with soft tissue sarcomas.
Immunohistochemical study of estrogen receptors in primary breast carcinomas and their lymph node metastases including comparison of two monoclonal antibodies
Authors: Nedergaard LOTTE , Haerslev TORBEN , Jacobsen GRETE KRAG
Published: August 2009

Estrogen receptors (ER) status was investigated in 101 primary breast carcinomas and their axillary lymph node metastases to determine if the malignant cells retained or changed this phenotypic feature during the metastatic process. Immunohistochemistry with the ER‐ICA kit (Abbott Laboratory) on formalin‐fixed paraffin‐embedded tissue was used (paraffin ER‐ICA). The ER status in primary and secondary tumours was concordant in 80 patients (79%) and discordant in 21 (21%). Eighteen of these twenty‐one patients had ER‐positive primary tumours and ER‐negative lymph node metastases. This discordance, which may be due to loss of ERs in the metastatic cells or tumour heterogeneity, could explain the well‐known failure of endocrine treatment in some of the patients with ER‐positive primary tumours. A new monoclonal antibody ID5 (DAKO) against ERs was applied on formalin‐fixed paraffin‐embedded tissue from 83 of these 101 primary carcinomas. These analyses and paraffin ER‐ICA analyses were compared to prior analyses of the same 83 tumours using the ER‐ICA kit on fresh frozen tissue (“gold standard”, frozen ER‐ICA). Kappa coefficient, sensitivity and specificity were 0.74, 0.96 and 0.75 for ID5 antibody, and 0.59, 0.72 and 0.96 for ER‐ICA antibody on paraffin sections.
The impact of the first notification of positive blood cultures on antibiotic therapy
Authors: SchØNheyder H. C. , HØJbjerg T.
Published: August 2009

The impact on antibiotic therapy of the first notification of positive blood cultures was assessed for 735 episodes of bacteraemia detected during 1992 in the County of Northern Jutland, Denmark. A primary focus of infection was defined in 498 episodes, the urinary tract being the most frequent (n = 182, 25%). Twenty‐nine patients (3.5%) had died prior to the initial contact. In 12 episodes antibiotic therapy had either been stopped or data were not available, leaving 694 episodes for further assessment. In 567 episodes antibiotic therapy had been started prior to the initial contact, the most frequent regimen being ampicillin or an ampicillin‐aminoglycoside combination (295 episodes), whereas cephalosporins, thienamycin, and fluoroquinolones were seldom used (41 episodes). The ongoing antibiotic coverage was deemed appropriate in 418 episodes (60%), non‐optimal in 90 (13%), and lacking in 186 (27%). The notification of positive blood cultures elicited changes in antibiotic therapy in 315 episodes (45%), including commencement of antibiotic therapy in 127 (18%). Thus, blood culture results have a measurable impact on antibiotic therapy.
Neuronal degeneration in subacute necrotizing encephalomyelopathy (Leigh's disease)
Authors: Lindboe C. F. , Lie A. K. , Aase S. T. , Schjetne O. B. , Haave I.
Published: August 2009

We report clinical, radiological and pathological findings in a 5‐year‐old girl who died of subacute necrotizing encephalomyelopathy (SNE) after 4 weeks of illness. Autopsy revealed endothelial swelling and vacuolar degeneration of the neuropil in the brain, brain stem and cerebellum. In addition, the affected areas showed degeneration of the neurons which was different from anoxic nerve cell damage both with regard to morphological picture and topographical distribution. This neuronal degeneration was probably due to the underlying metabolic defect in SNE per se and resembled in several aspects the nerve cell changes seen in the thalami and inferior olives in active Wernicke's encephalopathy. It is our opinion that more attention should be paid to the nerve cell degeneration in SNE rather than focusing on the relative preservation of these cells.
Antineutrophil cytoplasmic autoantibodies, autoantigens, and systemic vasculitis
Authors: Gross WOLFGANG L. , Csernok ELENA , Helmchen UDO
Published: August 2009

Antineutrophil cytoplasmic antibodies (ANCA) encompass a heterogeneous group of autoantibodies targeting antigens in neutrophils (PMN), monocytes, and endothelial cells. ANCA are routinely detected by the indirect immunofluorescence technique (IFT) and at least three different patterns of fluorescence can be distinguished which have been assigned the acronyms cANCA, pANCA and aANCA. cANCA is mostly induced by proteinase 3 (PR3) antibodies (PR3‐ANCA), and pANCA by myeloperoxidase (MPO) antibodies (MPO‐ANCA), while aANCA has unidentified subspecificity. Over the past decade, ANCA have been the subject of extensive investigation. They have proved to be of significant value both as diagnostic tools and for follow‐up in several forms of systemic vasculitis (e.g. Wegener's granulomatosis, WG; microscopic polyarteritis, MPA; Churg‐Strauss syndrome, CSS) which are now termed ‘ANCA‐associated vasculitides’. Furthermore, it is suspected that the presence of ANCA is an important factor in the pathogenesis of these disease groups. Data regarding the detection of ANCA and their diagnostic value and role in the pathogenesis of vasculitic disorders will be discussed in this review. Growing evidence points to a pathophysiological and diagnostic relevance of the distribution of the ANCA target antigens PR3 and MPO (presence in the circulation, on cellmembranes, and in tissue extracellularly). An autoimmune process has been implicated in the pathogenesis of ANCA‐associated vasculitis, but it is uncertain which mechanism underlies the induction of the ANCA‐related immunoresponse. In this paper mechanisms such as antigenic cross‐reactivity between human PMN proteins and extrinsic antigens by molecular mimicry, idiotypic immunoglobulin regulation, and T‐cell reactivity to PR3 and MPO will be discussed.
Inhibitory effect of lactoferrin on the adhesion of Actinobacillus actinomycetemcotnitans and Prevotella intermedia to fibroblasts and epithelial cells
Authors: Alugupalli KISHORE R. , Kalfas SOTIRIOS
Published: August 2009

Adhesion of the periodontitis‐associated bacteria Actinobacillus actinomycetemcomituns and Prevotella intermedia to monolayers of fibroblasts, HEp‐2, KB and HeLa cells was quantified with radiolabeled bacteria. Bacterial adhesion was also examined microscopically with Giemsa‐stained nonradioactive preparations. The degree of bacterial adherence was dependent on the growth phase of the bacteria. Strains at the exponential phase adhered to a greater extent than those at the stationary phase of growth. Both human and bovine lactoferrins competitively inhibited the adhesion of A. actinomycetemcomitans and P. intermedia to all tested cell monolayers. The inhibitory effect was dose‐dependent in the concentration range 0.5–2500 μg/ml and not related to the bacterial growth phase. In the presence of lactoferrin, decreased association of bacteria with the cell monolayers was also found by microscopic examination of the preparations. The present findings indicate that lactoferrin may prevent the establishment of bacteria in periodontal tissues through adhesion‐counteracting mechanisms in addition to its bacteriostatic and bactericidal properties.
Role of interferons and other cytokines in the regulation of the immune response
Authors: Belardelli FILIPPO
Published: August 2009

Cytokines represent the major factors involved in the communication between T cells, macrophages and other immune cells in the course of an immune response to antigens and infectious agents. A number of studies on mouse and human T helper (Th) clones have recently provided extensive evidence for the existence of different activities exhibited by Th cells (called Th1 and Th2), which was apparently inferred from the profile of cytokine secretion. The Thl‐type immune response is generally associated with IgG2a production and the development of cellular immunity, the Th2‐type response with IgE production, eosinophils and mast cell production. This review focuses on the role of different cytokines produced by macrophages (especially interferons (IFNs), TNF‐α, IL‐10 and IL‐12) or T cells (IFNs, IL‐2, IL‐4, IL‐10, IL‐13 and TGF‐β) in macrophage‐T cell interactions and the cytokine relevance in the differentiation of Th cells towards the Thl or Th2 type of immune response. Th1‐derived cytokines (IFN‐gamma, IL‐2, TNF‐α) favor macrophage activation, whereas the Th2 cytokines (IL‐4, IL‐10, IL‐13) exhibit suppressive activities on macrophage functions. A key role in the differentiation towards the Th1‐type response is now attributed to IL‐12, a recently described cytokine produced mainly by macrophages. Its production can be upregulated by IFN‐gamma and is inhibited by IL‐10 and IL‐4. All this emphasizes the importance of macrophage‐cytokine interactions in determining the type of immune response. This article also aims to review recent data concerning the roles of IFNs α/β (type I) and IFN‐gamma (type II) in the regulation of the immune response. While there is much information on the regulatory effects of IFN‐gamma (also called “immune IFN”) on the immune response, little is so far known of the role of type I IFNs. These cytokines, originally described as simple antiviral substances, are now taken to be important regulators of the immune response. Recent data indicate that these molecules (especially IFNs‐α) specifically promote the differentiation towards the Th1‐type response. The stimulatory effects of IFN‐α on the generation of the Th1‐type response may be involved in its therapeutic effects in some human diseases, including early AIDS, hypereosinophilia and certain tumors. It is reasonable to assume that, with the increasing interaction between basic and clinical research, considerably more will be understood about how IFNs and other cytokines interact in the modulation of the immune response, and how this knowledge can be successfully translated into new and more selective therapeutic strategies against human diseases.
Chromosomal restriction endonuclease analysis and ribotyping of Bacteroides fragilis
Authors: Kleivdal H. , Hofstad T.
Published: August 2009

We have analysed the restriction fragment length patterns of chromosomal DNA and of ribosomal RNA (rRNA) genes in order to investigate the clonal distribution within Bacteroides fragilis isolates. Eighteen blood culture isolates from 18 patients and 4 faecal isolates from 4 subjects were examined. Chromosomal restriction endonuclease analysis (REA) was performed by separating BamHI‐gener‐ated DNA fragments using polyacrylamide gel electrophoresis (PAGE). Ribotyping was accomplished by hybridizing EcoRI‐treated genomic DNA subjected to conventional REA on agarose to a radio‐labelled probe obtained from 16+23S rRNA of E. coli. All 22 isolates could be differentiated by their REA patterns with a varying percentage of similar fragments. Analysis of the rRNA gene patterns displayed heterogeneity, and revealed 14 ribotypes among the 18 blood culture isolates and 3 among the 4 faecal isolates. The predominant ribotype among the clinical isolates was also shared by one faecal isolate. The results suggest that no particular clones are predominantly responsible for systemic infection.
Localization of NCAM on NCAM‐B‐expressing cells with inhibited migration in collagen
Authors: Meyer MALENE B. , Bastholm LONE , Nielsen MORTEN H. , Elling FOLMER , Rygaard JØRGEN , Chen WEICHING , ÖBrink BJÖRN , Bock ELISABETH , Edvardsen KLAUS
Published: August 2009

The extracellular matrix is a key element in neuronal development and tumour invasion, providing a substratum which sustains the adhesion and migration of cells. In order to study interactions between the neural cell adhesion molecule (NCAM) and collagen, we transfected mouse L cells with cDNA encoding the human transmembrane NCAM isoform of 140 kDa (NCAM‐B). An L‐cell/collagen type I system was used to study the influence of NCAM expression on in vitro invasion. We here report that migration of NCAM‐expressing cells in collagen was inhibited compared to that of NCAM‐negative cells transfected with the empty vector. Immunofluorescence confocal laser scanning microscopy (CLSM) and immunogold electron microscopy using anti‐human NCAM antibodies demonstrated a heterogeneous distribution of NCAM on the plasma membrane of transfected L cells grown on collagen. NCAM was preferentially located at the surface of broad cytoplasmic protrusions and slender extensions, some of which were facing the collagen. This was in contrast to the homogeneous surface distribution of NCAM on cells grown on plastic. These data suggest that NCAM and collagen type I interact, and that this might lead to the migration inhibition of NCAM‐expressing cells.
Comparison of tests designed to identify Staphylococcus aureus thermostable nuclease
Authors: Brakstad ODD G. , MæLand JOHAN A. , Chesneau OLIVIER
Published: August 2009

The thermonuclease test has been employed for same‐day identification of Staphylococcus aureus in blood cultures and a seroinhibition test to confirm S. aureus thermonuclease (TNase) by using polyclonal anti‐S. aureus TNase antiserum. However, strains of non‐S. aureus staphylococci may produce TNases which are neutralized by the antiserum. This study evaluated alternative reagents and confirmatory tests for the S. aureus TNase. The tests included seroinhibition by a monoclonal antibody (MAb) against S. aureus TNase, MAb‐based detection of the TNase in a sandwich ELISA, and a polymerase chain reaction for amplification of the nuc gene encoding the S. aureus TNase. All these tests discriminated between TNase produced by S. aureus and TNase produced by strains of the species S. caprae, S. carnosus, S. simulans, S. capitis, and S. intermedius. Specificity for the S. aureus TNase was confirmed by an inhibition ELISA for two out of three MAbs tested. Thus, the MAb‐and nwobased tests will be useful to discriminate between TNases from S. aureus and non‐S. aureus staphylococci.
erm genes in erythromycin‐resistant Staphylococcus aureus and coagulase‐negative staphylococci
Authors: Westh H. , Hougaard D. M. , Vuust J. , Rosdahl V. T.
Published: August 2009

Erythromycin‐resistant S. aureus from general practice and a dermatology ward with a very high consumption of erythromycin was studied for ermA and ermC content by Southern blot analysis. The prevalence of these erm genes was also studied in coagulase‐negative staphylococci from the same dermatology ward and in a collection of 15 S. aureus and 18 coagulase‐negative staphylococci found in the same specimen from 15 different patients. ermA was only found as a chromosomal insert and ermC only on small plasmids. In erythromycin‐resistant S. aureus from general practice ermC was responsible for 84% of erythromycin resistance, while 16% of the strains contained ermA. In 17 of 18 S. aureus strains from the dermatology ward a 2.5 kb plasmid contained ermC. Among 58 erythromycin‐resistant coagulase‐negative staphylococci only 2 (3%) had ermA, whereas 44 strains (76%) had ermC. ermA or ermC genes were common in S. epidermidis (36/38). However, in 20 isolates of erythromycin‐resistant coagulase‐negative staphylococci of other species, 10 had neither ermA nor ermC. ermC was the most common erm gene in both coagulase‐negative staphylococci and S. aureus. In 11 of 15 patients with erythromycin‐resistant S. aureus the co‐isolated erythromycin‐resistant coagulase‐negative staphylococcus had another resistance mechanism, either another gene or a different pheno‐typic expression of the same gene. Resistance to pristinamycin, a streptogramin antibiotic only used in animals, was not found in S. aureus but was found in 14% of erythromycin‐resistant coagulase‐negative staphylococci.
Effects of calprotectin in avridine‐induced arthritis
Authors: Brun JOHAN G. , HÅLand GEIR , Haga HANS‐JACOB , Fagerhol MAGNE K. , Jonsson ROLAND
Published: August 2009

Plasma levels of calprotectin correlate with disease activity and clinical assessments of arthritis in various rheumatic diseases, and high levels have been demonstrated in the synovial fluid of patients with rheumatoid arthritis. However, the role of calprotectin in rheumatic inflammation is unclear. The purpose of the present study was to investigate potential intra‐articular effects of calprotectin. Calprotectin was injected into joints of healthy male Lewis rats and into joints of rats in the latency period before onset of avridine‐induced arthritis. In addition, a group of animals had IgG antibodies to rat calprotectin injected into joints before onset of avridine‐induced arthritis. Injection of 0.2 or 10 μ calprotectin into the ankles of healthy male Lewis rats resulted in histologically minor and reversible inflammatory changes, but without any circulating antibodies to calprotectin. Furthermore, animals with 40 μg calprotectin injected into ankles before the expected onset of avridine‐induced arthritis had lower scores for cellular infiltration than were seen in control joints. This difference did not quite reach statistical significance in the two‐sided test used. However, the induced arthritis increased in joints injected with IgG antibodies to calprotectin. These findings may indicate that increased local concentrations of calprotectin are partially protective against avridine‐induced arthritis. In contrast, reduced local concentrations appear to exacerbate the severity of arthritis. Calprotectin may thus be involved in the regulation of inflammatory processes in joints.
Changes in lymphocyte subpopulations and adhesion/activation molecules following endotoxemia and major surgery
Authors: Toft P. , Hokland M. , Hansen T. G. , TØNnesen E.
Published: August 2009

Major surgery as well as endotoxin‐induced sepsis is accompanied by lymphocytopenia in peripheral blood. The purpose of this study was to investigate the redistribution of lymphocyte subpopulations and adhesion/activation molecules on lymphocytes. Twenty‐four rats were included in the investigation. Eight rats received an intraperitoneal injection of E. coli endotoxin (2 mg kg−1), eight rats had a sham operation performed while eight rats received isotonic saline and served as a control group. Blood samples were obtained by making an incision in the tail before and 2 and 5 h after surgery or administration of endotoxin or saline. After isolation of lymphocytes by gradient centrifugation, flow‐cytometric immunophenotyping was performed using CD2, CD3, CD4, CD8, CD11/CD18, CD20, CD44 and MHC II monoclonal antibodies. Endotoxemia and surgery were both accompanied by increased serum Cortisol, lymphocytopenia, and a decrease in CD2, CD3 and CD4 lymphocytes. Only endotoxemia was followed by a decrease in CD8, CD11/CD18 and CD44 lymphocytes in peripheral blood. Our results show that several of the changes in lymphocyte subpopulations following surgery and sepsis are associated with increased serum Cortisol. Sepsis is, in addition, accompanied by an upregulation of adhesion receptors.
Trophozoite‐to‐cyst ratio increases during recovery from Pneumocystis carinii pneumonia in rats
Authors: Sukura ANTTI
Published: August 2009

Pneumocystis carinii has two developmental stages: the trophozoite and the cyst. The differential specificity of monoclonal antibodies SMoAb (cysts only) and CMoAb (cysts and trophozoites) was used to analyse the trophozoite‐to‐cyst (T : C) ratio during recovery from P. carinii pneumonia (PCP) in a dexamethasone model in Wistar rats. In tissue homogenates the cysts decreased from week 0 to week 4 (4.10±1.57 106/g vs 0.069±0.021 106/g, p<0.001), whereas the trophozoites decreased much less, leading to a change in T: C ratio from week 0 and 1 (72±33 and 49±14, respectively) to week 2 and 4 (1752±652 and 1163±482, p<0.001). A similar tendency was found in immunofluorescence‐labelled histological samples quantitated with the VIDAS image analyser. These results demonstrate a hitherto unrecognized and significant change in T : C ratio during recovery from PCP. This change could not be monitored using bronchoalveolar fluid, probably due to adherence of the trophozoites to the alveolar wall. Changes in T : C ratio indicate that in particular the cyst form of P. carinii is sensitive to the host response mounted during recovery, and support the theory that trophozoites probably also multiply in an extracystic asexual fission.
c‐erbB‐2 oncoprotein in relation to DNA ploidy and prognosis in colorectal adenocarcinoma
Authors: Sun XIAO‐FENG , Carstensen JOHN M. , StÅL OLLE , Zhang HONG , NordenskjÖLd BO
Published: August 2009

Overexpression of the c‐erbB‐2 onocogene has been demonstrated in a variety of tumours, including colorectal tumours. In breast carcinoma, c‐erbB‐2 overexpression is associated with DNA ploidy, some other prognostic indicators, and unfavourable survival prospects. However, there is little such information available regarding colorectal tumours. In this study, c‐erbB‐2 was analysed retrospectively by immunohistochemistry in 293 primary colorectal adenocarcinomas to assess its relation to DNA ploidy, S‐phase fraction, other prognostic factors, and patient survival. Using the monoclonal antibody NCL‐CB11, we found that 23% of the tumours were strongly c‐erbB‐2 positive, while 36% showed weak expression. The highest frequency of c‐erbB‐2 expression was 81% in DNA tetraploid tumours, compared to 63% in aneuploid and, 53% in diploid tumours (test for heterogeneity, p=0.031). Overexpression of c‐erbB‐2 indicated a favourable prognosis in patients with DNA aneuploid tumours (p=0.0088), but not in those with diploid or tetraploid tumours. The prognostic value of c‐erbB‐2 in DNA aneuploid tumours remained even after adjustment for Dukes' stage (p=0.027). The results suggest that a combination of c‐erbB‐2 expression and DNA ploidy may improve the identification of patients' risk of cancer death.
Helicobacter pylori‐induced lipid peroxidation in peripheral blood lymphocytes
Authors: Fan XUE G. , Yakoob JAVED , Chua ANDREW , Fan XUE J. , Keeling PAUL W. N.
Published: August 2009

Several lines of evidence implicate Helicobacter pylori infection in the pathogenesis of gastritis and peptic ulceration. To investigate whether H. pylori can cause lipid peroxidation in lymphocytes in vitro and to look for experimental evidence of lipid peroxidation induced by H. pylori, the lipid peroxide (LPO) level in peripheral blood lymphocytes was measured using the thiobarbituric acid fluorescence method. In the absence of added H. pylori, the LPO level in lymphocytes was 0.133±0.033 nmol/106 cells, and in the co‐culture of H. pylori with peripheral blood mononuclear cells 0.340±0.097 nmol/ 106 cells. A significant difference was found between the two groups (p<0.001). Antioxidants, either superoxide dismutase or catalase, could inhibit LPO production in lymphocytes. The present data provide further evidence that H. pylori can induce lipid peroxidation, which may be responsible for the pathogenesis of H. pylori‐associated mucosal damage.
New approaches to hepatitis A and B vaccines
Authors: Iwarson STEN
Published: August 2009

Plasma‐derived vaccines and yeast‐derived recombinant vaccines against hepatitis B virus (HBV) infection have gained an acceptable record of efficacy. However, non‐ or hyporesponsiveness to immunization does not only occur in cases of obesity, renal failure or immune suppression, but also in healthy individuals. There is therefore a rationale for developing more immunogenic vaccines against HBV especially for those populations who are potential non‐ or hyporesponders. Currently used recombinant hepatitis B vaccines consist of antigen particles assembled with the product of 226 amino acids encoded in the S gene. Since proteins encoded in the pre‐S gene are also incorporated in the HBV envelope, pre‐S gene products should, at least in theory, be useful in improving protection with hepatitis B vaccines. Inactivated hepatitis A vaccines are more potent than currently used hepatitis B vaccines. Two injections of a standard dose of HAVRIX (SB) by the intramuscular route, or even a single injection using a higher dose (HAVRIX 1440), will achieve protective levels of antibodies. Therefore, increased potency is not essential with inactivated hepatitis A vaccines. New hepatitis A vaccines are likely to be recombinant or attenuated live types. Another aspect of the improvement of existing hepatitis A and B vaccines is unification into a combined form.
Seasonal variations in the occurrence of environmental mycobacteria in potable water
Authors: Kubalek IGOR , Komenda STANISLAV
Published: August 2009

A fluctuation in the prevalence of environmental mycobacteria in relation to nutritional conditions in nature has been repeatedly described in the literature. A seasonal difference in the potable water supply system has not yet been documented. Potable water samples from the supply systems of 16 identical localities were analyzed. Samples of running water and tap swabs or tap scrapings were collected twice a year, in the spring and in the autumn. The samples were processed as stipulated by the international standards. McNemar's test was used to analyze the difference in the occurrence of environmental mycobacteria between the vernal and the autumnal samples. A significant change in the presence of environmental mycobacteria in the potable water supply system was observed, the vernal samples yielding more positive results, This finding supports other observations respecting surface water. We suggest that this effect on the potable water supply system may be caused by the change in temperature. Contamination rates were similar, with no statistically significant differences between running water samples and those from swabs or scrapings. No time trend in the perod 1984–1989 respecting the prevalence of mycobacteria was detected. Direct microscopy showed massive colonization with environmental mycobacteria of the potable water supply system. The public health consequences of these findings should be further evaluated, as colonization of water pipes can be associated with outbreaks of mycobacterial disease in immunocompromised patients. There has also been an increase in the incidence of mycobacterioses in the North Moravian region in recent years.
An improved, sensitive, non‐radioactive in situ hybridization method for the detection of cytokine mRNAs
Authors: Klein SIGRID C. , VAN Wichen DICK F. , Golverdingen JOKE G. , Jacobse KAREN C. , Broekhuizen ROEL , DE Weger ROEL A.
Published: August 2009

We established an improved non‐radioactive in situ hybridization (ISH) method to detect mRNA of cytokines in cell preparations and tissues. Via this method we could demonstrate various cytokines in stimulated peripheral blood mononuclear cells (PBMC), lymphoid cell lines and human lymphoid tissues. The probes for the in situ hybridization were made by labelling cytokine‐specific PCR products with digoxigenin (Dig) in a repeated PCR. This resulted in an intrinsic labelling of the probe with several Dig‐UTP molecules. Incorporation of Dig‐11‐dUTPs was shown on ethidium bromide‐stained agarose gels by a higher molecular weight of the PCR products with incorporated Dig‐dUTPs when compared to control PCR products without digoxigenin. Cytospin‐centrifuged cells of PHA‐stimu‐lated PBMC or lymphoid cell lines and frozen sections of various human lymphoid tissues were hybridized with the Dig‐labelled cytokine probes and the hybridized probes were detected immuno‐histochemically. In this way, we detected and localized cytokine mRNAs (IL‐2, IL‐4, IL‐6, IL‐8, IL‐10) in PBMC, in the human T‐cell line Jurkat, in the follicular lymphoma cell line DoHH2, and in human lymph nodes and tonsils. The in situ hybridization had a high sensitivity as the results correlated closely with the detection of cytokine mRNA by reverse transcriptase‐PCR (RT‐PCR) data from the same samples. We showed that Jurkat and DoHH2 cells produce several cytokines constitutively and that, after activation with the phorbol ester PMA, expression of several cytokine mRNAS was enhanced.
The glomerular mesangium: studies of its developmental origin and markers in vivo and in vitro
Authors: HÖLthofer HARRY , Sainio KIRSI , Miettinen AARO
Published: August 2009

Cultured mesangial cells are widely used to explore their role in kidney glomerular functions, but methods to reliably identify these cells in vivo and in vitro are lacking. Furthermore, the proposed relationship of mesangial cells to e.g. fibroblasts and smooth muscle cells has not been systematically studied. Here we wanted to search for markers of practical use also in identifying cultured mesangial cells, and to apply these markers in a study of the origin of glomerular mesangium. No epitopes specific for only mesangial cells could be identified, and no evidence of their true relationship with neural or lymphocytic lineages could be found. Findings with the variety of markers used suggest that mesangial cells may be indistinguishable from smooth muscle cells and fibroblasts. A panel of antibodies, including those against Thy 1.1, smooth muscle actin, desmin, cellular fibronectin and pi integrin al and a5 chains, and Wistaria floribunda (WFA) and Ricinus communis (RCA I) lectins, were found useful for mesangial cell detection in vivo and in vitro. The origin of glomerular mesangial cells could not be conclusively determined, although the results indirectly suggest that mesangial cells together with endothelial cells migrate to the glomerulus from the outside.
Antimicrobial screening of zinc in the absence or presence of oleoresins and various resin acids
Authors: Johansson ANDERS , Sunzel BO , Holm STIG E. , SÖDerberg THOR , Gref ROLF
Published: August 2009

Zinc and oleoresins are the main components of several wound dressings, and are also frequently used in root canal treatment. The in vitro antibacterial effects of zinc, six highly purified resin acids and two commercial oleoresins alone or combined in varying proportions were analysed. Oleoresins are composed of approximately 90% resin acids and the most common acids were included in this study. The antibacterial activity of the various chemicals was estimated using a Bioscreen robot analyser, which allowed 24 h kinetic documentation of bacterial growth. The bacteria employed were reference species commonly occurring on human skin or of oral origin. Zinc as well as the oleoresins and the pure resin acids all showed antibacterial activity when present in growth media, but the sensitivity of the bacteria varied. The presence of resin acids and oleoresins increased the antibacterial effect of zinc to varying degrees depending on the combination and the bacterial species tested. The results of the present study indicate that zinc, resin acids, or oleoresins alone, as well as combined, show antibacterial activity against selected aerobic Gram‐positive and anaerobic Gram‐negative bacteria.
Fcγ‐receptor activity in the developing human placenta
Authors: Jensen T. SKEIE , Matre R.
Published: August 2009

The expression of Fey receptors (FcγR) and annexin II in 20 placentae (range 8–27 weeks' gestation) and in 3 full‐term placentae was studied using monoclonal antibodies (mAbs) and soluble immune complexes. Functionally active FcyR was detected on the trophoblast, endothelial cells, and stromal cells (Hofbauer cells) from the 8th week of gestation. FcγRI (mAb 32.2) and FcγRIII (mAb 3G8) were detected only on Hofbauer cells, whereas FcγRII (IV3 and C1KM5) were detected both on Hofbauer cells and endothelial cells. FcγRIII (anti‐Leu‐lib) and the IgG‐binding molecule annexin II (mAb B1D6) were expressed by Hofbauer cells, endothelial cells, and the trophoblast. There was some variation in staining among the different specimens, but the number of positive cells as well as the staining intensity increased from the first to the second trimester. In first trimester placenta, staining was localized both to the syncytio‐ and cytotrophoblast, with the strongest intensity in the cytotrophoblast and at the boundary between the two cell layers. In second and third trimester placenta, staining was localized to the syncytiotrophoblast. The localization and distribution of FcγR on the trophoblast during ontogeny is of interest with regard to its presumed role in the transport of IgG from mother to fetus.
Comparison of adherent and non‐adherent staphylococci in the induction of polymorphonuclear leukocyte activation in vitro
Authors: Riber ULLA , Espersen FRANK , Kharazmi ARSALAN
Published: August 2009

The ability to consume complement and activate neutrophils was investigated for staphylococci adherent to silicone surfaces and non‐adherent staphylococci. Staphylococcus epidermidis strain ATCC 14990 and Staphylococcus aureus strain E 2371 were used in this study. The bacteria were allowed to adhere to silicone catheter segments for 2 h at 37 °C. Complement consumption was measured by reduction in serum haemolytic activity against sheep red blood cells. The induction of chemilumi‐nescence was measured after opsonization of the staphylococci in 20% AB‐positive human serum for 60 min at 37 °C. The bacteria consumed complement to approximately the same extent when adherent to the catheter segments, but more slowly in comparison with planktonic bacteria. When planktonic bacteria were compared, complement was consumed more quickly by S. epidermidis than by S. aureus. Measuring the induction of chemiluminescence by planktonic bacteria, S. epidermidis induced a lower response than S. aureus, while when adherent to the catheter segments the bacteria induced similar responses. These responses were only 15 to 20% of those induced by planktonic bacteria and only slightly higher than the spontaneous chemiluminescence by the neutrophils. Inter‐strain variation was found, but all strains induced about the same low chemiluminescence when adherent to the catheter segments. The reduction in inflammatory response caused by adherence of staphylococci to catheter segments may interfere with phagocytosis and elimination of S. epidermidis during the early establishment of a foreign body infection.
p53 immunostaining in combined type small cell lung cancer
Authors: Higashiyama MASAHIKO , Doi OSAMU , Yokouchi HIDEOKI , Kodama KEN , Tateishi RYUHEI
Published: August 2009

Abnormalities of the p53 oncogene in lung cancer have recently been reported to be more frequent in small cell lung cancer (SCLC) than in non‐small cell lung cancer (non‐SCLC), but their status in combined type SCLC is as yet unknown. In this study, immunohistochemical analysis using a polyclonal antibody against p53 protein was performed in 12 surgically resected specimens of combined type SCLC. Immunoreactivity of the p53 protein was found in 5 (42%) of the 12 cases, and the immunostaining pattern of the p53 protein in areas of the non‐small cell carcinoma type was the same as in those of the small cell carcinoma type. Thus, it seems that the incidence of p53 abnormalities in combined type SCLC is slightly lower than in ordinary type SCLC. It is also suggested that abnormalities of the p53 oncogene in this histological type may not be a specific event related to the morphological difference between small cell carcinoma and non‐small cell carcinoma.