bluuebunny/crossref_metadata_2025 huggingface dataset:
The indoor microfungusTrichoderma viridepotentiates histamine release from human bronchoalveolar cells
Authors: Larsen F. O. , Clementsen P. , Hansen M. , MaltbæK N. , Gravesen S. , Skov P. Stahl , Norn S.
Published: August 2009

Trichoderma viride(Tv) is often found in damp and mouldy buildings where people complain of adverse health effects including mucosal/respiratory symptoms. Inhaled spores can reach the alveoli and may interact with the airway epithelium. An interaction with the mucosal mast cells was studied in cells obtained by bronchoalveolar lavage (BAL) from 18 individuals. The fungal spores were found to trigger histamine release from the BAL cells, but relatively high concentrations (0.1–2 mg/ml) were needed. A similar dose response was obtained in basophil histamine release. The Tv‐induced mediator release was caused by non‐immunological (non‐IgE‐dependent) mechanisms since the histamine release was not changed by removal of IgE from the basophils before exposure of the cells to the spores. However, in very low concentrations (0.1 ng/ml) the fungal spores were found to potentiate IgE‐mediated histamine release triggered by anti‐IgE antibody in suspensions of BAL cells. Potentiation was also obtained in basophil histamine release, but relatively high concentrations of Tv (10−2mg/ml) were needed. Ourin vitroexperiments show that mucosal mast cells from the airways are highly sensitive to the potentiating effect of Tv. Although inhalation studies are needed to determine thein vivoeffect of the spores, the results suggest reinforcement of mediator release to be a mechanism in the adverse health implications observed in mouldy buildings.
B‐ and T‐lymphocyte subpopulations in the adenoids of children with otitis media
Authors: Mattila Petri S. , Tarkkanen Jussi
Published: August 2009

Adenoid hyperplasia is connected with the pathogenesis of otitis media in children, as implicated by the residence of pathogens causing otitis media in adenoids, and by the beneficial effect of adenoidectomy. We have quantitated the proportions of certain T‐ and B‐lymphocyte subsets in the adenoids by three‐color flow cytometry. Three groups were studied: 1) children with persistent effusion in the middle ear (pOME), 2) children with recurrent otitis media with no effusion in the middle ear at the time of surgery. (rAOM), and 3) children with adenoid hyperplasia without otitis media (AH) presenting either with snoring or upper respiratory tract infections. B lymphocytes comprised on average 64, 66 and 58% of all mononuclear cells in the respective patient groups. The majority of the B lymphocytes were of the follicular mantle zone phenotype expressing surface IgD (on average 69, 67 and 70% of all B lymphocytes in the respective patient groups). Almost one third of the B lymphocytes were of the germinal center cell phenotype, expressing CD38 but negative for surface IgD (on average 29, 30 and 25% in the respective patient groups). CD3+T lymphocytes comprised 33, 33 and 40%, respectively, of all mononuclear cells, and the average CD4+/CD8+ratio was 5.2, 4.9 and 5.7, respectively. No statistically significant differences between the patient groups were seen in these T‐ and B‐lymphocyte subpopulations. These results suggest that the relative sizes of the germinal center, follicular mantle and interfollicular compartments of the adenoids do not correlate with the disease status of the middle ear cavity.
Effect of insulin and epidermal growth factors on the synthesis of glycosaminoglycans/proteoglycans in cultured human malignant mesothelioma cells of different phenotypic morphology
Authors: Tzanakakis George N. , Karamanos Nikos K. , Syrokou Alexandra , Hjerpe ANders
Published: August 2009

Two human malignant mesothelioma cell sublines, one with a fibroblast‐like and the other with an epithelial differentiation, were examined for their capacity to synthesize glycosaminoglycans in the presence of IGF‐I, EGF, and their combination. This synthesis depends on the morphology of the mesothelioma cells, with many‐fold higher amounts of both hyaluronan and proteoglycans being produced by the cells with epithelial morphology than by those of the fibroblast phenotype. In both cell lines this synthesis was affected in a dose‐dependent fashion by the exogenously added growth factors and exposure to IGF‐I and EGF in combination showed a synergistic effect. This effect of those factors seems to be mediated via protein tyrosin kinase‐dependent receptors and was different in the fibroblast‐like and epithelial cells. The synthetic rates of the various glycosaminoglycans formed (hyaluronan, galactosaminoglycans and heparan sulfate) were also variously affected by these factors, indicating differences in how the synthesis of the various glycosaminoglycans is regulated. The results obtained suggest a close correlation between the presence of the appropriate growth factor(s), the process of cell differentiation and the synthesis of glycosaminoglycans in these cells.
Antibody reactivities to glutamate‐rich peptides of Plasmodium falciparum parasites in humans from areas of different malaria endemicity
Authors: Jakobsen P. H. , Theander T. G. , Hviid L. , Morris‐Jones S. , Jensen J. B. , Bayoumi R. A. L. , Greenwood B. M. , Bygbjerg I. C. , Heegaard P. M. H.
Published: August 2009

Synthetic P. falciparum peptides were evaluated as tools in epidemiological investigations of malaria. Plasma IgM and IgG antibody reactivities against synthetic peptides covering sequences of glutamate‐rich protein (GLURP) and acidic‐basic repeat antigen (ABRA) were measured by ELISA in individuals from malaria‐endemic areas of Sudan, Indonesia and The Gambia to study antibody responses to these peptides in donors living in areas of different malaria endemicity. IgG and IgM reactivities to the peptides increased with malaria endemicity, although there were no differences in reactivities to the GLURP peptide between non‐exposed donors and donors living in areas of low malaria endemicity. IgG reactivities to the GLURP peptide in Sudanese adults were high one month after treatment in all adults tested, while IgG reactivities to the ABRA peptide were infrequent. IgM responses to the peptides tested were shortlived in most patients. In Gambian children with malaria, IgM reactivities but not IgG antibody reactivities against the ABRA peptide were higher in those with mild malaria than in those with severe malaria. The peptides may be useful in future epidemiological studies, especially in areas of low malaria endemicity.
Histo‐blood group antigens in human fetal thymus and in thymomas
Authors: Engel Peter , Dabelsteen Erik , Francis Dorthe , GræM Niels
Published: August 2009

The glycosylation of epithelial cell surface antigens follows cellular differentiation, and changes in the pattern of expression are seen in various premalignant and malignant epithelial lesions. The distribution of type‐2 chain ABH‐carbohydrate structures (N‐acetyl‐lactosamine, H‐type 2 chain, Le‐y, Le‐x and sialyl‐Le‐x) of the ABO‐histo‐blood group system was investigated in 19 normal fetal thymuses (gestational age 16 to 39 weeks) and in 19 thymomas in order to study possible tumor‐associated changes in the glycosylation pattern. The material was investigated by immunochemical stainings of formalin‐fixed paraffin‐imbedded tissue using monoclonal antibodies with defined specificity. In fetal thymus the epithelial cells of the medulla and the Hassal's bodies strongly expressed elongated carbohydrate structures (Le‐y, Le‐x and sialyl‐Le‐x). In a few cases the cortical epithelial cells weakly expressed Le‐x and sialyl‐Le‐x. Compared with fetal thymus 16 of the thymomas showed a total loss, or a very much reduced expression of elongated carbohydrate structures. Three thymomas, which histologically had been reclassified according to Kirchner & Müller‐Hermelink (14) as high grade thymic carcinomas, revealed strong expression of Le‐y, moderate expression of Le‐x and weak expression of sialyl‐Le‐x. This is of interest as in other tumors Le‐y is correlated with increased cell motility and with poor prognosis.
Microbiologically verified diagnosis of infectious spondylitis using CT‐guided fine needle biopsy
Authors: Rieneck K. , Hansen S.‐E. , Karle A. , Gutschik E.
Published: August 2009

Using a computed tomography (CT)‐guided technique we have been able to obtain fine needle spine biopsies directly from an affected vertebra or disk plate in 14 patients suspected of infectious spondylitis. The bioptic material was cultivated immediately and incubated for 14 days. Cultures from eight patients were positive. No single microbiological agent was predominant though coagulase‐negative staphylococci were frequent. In no case were mycobacteria found. Bioptic material from six patients did not give rise to growth of microorganisms. We were able to successfully treat the eight patients with a culture‐positive biopsy. We think that biopsies are crucial for establishing a microbiological diagnosis. The whole procedure takes less than one hour; it is performed under local anaesthesia and is thus not very stressful for the patient: The success rate for obtaining a positive spine biopsy was 57%.
Aminoglycoside resistance mechanisms in Enterobacteriaceae and Pseudomonas spp. from two Danish hospitals: correlation with type of aminoglycoside used
Authors: Busch‐SØRensen Charlotte , SØNmezoglu Meral , Frimodt‐MØLler Niels , HØJbjerg Tove , Miller George H. , Espersen Frank
Published: August 2009

Sixty‐two aminoglycoside‐resistant Gram‐negative enteric bacteria were isolated over a 3‐year period from two hospitals (Bispebjerg and Esbjerg) among a total of almost 270,000 isolates. These hospitals were selected because of their different aminoglycoside policies during the years investigated. At Bispebjerg Hospital the principal aminoglycoside used was tobramycin, while gentamicin was the first choice at Esbjerg Hospital. Escherichia coli was the most frequently found aminoglycoside‐resistant species. Among the 61 aminoglycoside‐resistant strains studied, resistance was due to aminoglycoside‐modifying enzymes in all except two Xanthomonas maltophilia strains. The ANT(2“) enzyme occurred significantly more often at Esbjerg Hospital (p=0.001), while enzymes of the AAC(3) or AAC(6‘) moieties were more common, but not significantly so, at Bispebjerg Hospital. The phenotypic pattern of aminoglycoside resistance, as determined by disc diffusion, correlated 100% with the ANT(2”) and AAC(3)‐V (the two most common enzymes among the isolates) genotype of the organisms as established using DNA probes. Median minimum inhibitory concentrations (MICs) (mg/1) for clinically utilized aminoglycosides were: amikacin (1.6), gentamicin (25.0), kanamycin (50.0), netilmicin (1.6–25.0) and tobramycin (12.5–50.0). Isolates from Bispebjerg Hospital revealed significantly higher MICs for netilmicin and tobramycin (p<0.01) as compared to isolates from Esbjerg Hospital.
Antigenic determinants in type 1 diabetes mellitus:
Authors: Dotta F. , Di Mario U.
Published: August 2009

Type 1 diabetes mellitus (IDDM) is a disease caused by the autoimmune destruction of insulin‐producing pancreatic ß cells that takes place in genetically predisposed individuals. The results of the studies performed so far during the search for “the target antigen” in ß cell autoimmunity have indicated that, unlike many autoimmune disorders, type 1 diabetes appears to be the result of an autoimmune response to a multiplicity of autoantigens. Autoantibodies and autoreactive T lymphocytes reacting with islet target molecules of protein or glycolipid nature have been shown in the circulation of individuals and of animal models of type 1 diabetes (NOD mouse and BB rat) before and at the onset of the disease. In the present article we have reviewed the data available on the antigenic determinants in type 1 diabetes, with particular reference to those recognized by autoantibodies which represent the best available predictive marker of future disease development in large scale screening studies.
Expression of tumor‐related antigens Lewisa, Lewisb, X, Y, Span‐1 and CEA in relation to differentiation and prognosis in rectal adenocarcinomas
Authors: Sun Xiao‐Feng , Zhang Hong
Published: August 2009

Expression of Lea (Lewisa), Leb (Lewisb), X (Lewis X), Y (Lewis Y), Span‐1 and CEA was detected by an immunohistochemical method using a panel of antibodies in paraffin‐embedded materials from 45 rectal adenocarcinomas and the corresponding normal mucosa. The relationships between antigen expression and grade of differentiation and survival were analyzed. Compared to the expression in normal mucosa, the expression of Lea was decreased in tumors while the expression of Leb, X, Y. Span‐1 and CEA was increased. The expression of Lea and Span‐1 was associated with the grade of differentiation (p=0.001 and p=0.02, respectively). Positive expression of Lea and Span‐1 correlated with poor prognosis (p=0.0001 and p=0.002, respectively), but positive expression of Y predicated favorable prognosis (p=0.01). Our findings indicate that some of the tumor‐related antigens might provide information important for the diagnosis, determining histologic differentiation and prognosis in rectal adenocarcinomas.
Recovery of skeletal muscle after immobilization of rabbit hindlimb:
Authors: Kauhanen Susanna , Leivo Ilmo , PettilÄ Markus , Michelsson JArl‐Erik
Published: August 2009

The histopathology of the deep portion of the vastus intermedius (VIP) muscle from 13 rabbit hind‐limbs immobilized in shortened position for 2–2.5 (n=4), 3 (n = 3), 14 (n = 3) and 28 (n = 3) days and VIP muscles from 13 rabbit hindlimbs immobilized for the same time periods and subsequently remobilized for 4 weeks were compared. After 3 days of immobilization the VIP muscles displayed a 15% (p<0.004) decline in muscle fibre diameter. By 2 weeks of immobilization fatty change was prominent and muscle fibre diameters had decreased to 56% (p<0.0001) of control values. By 4 weeks of immobilization severe fibrotic damage of myofibres was observed and fibre diameters had decreased to 47% (p<0.0001) of control values. Three days of immobilization followed by 4 weeks of remobiliza‐tion led to marked fatty change as well as an increase in connective tissue in the affected VIP muscles. The muscle fibre diameter in these muscles was 71% (p<0.0001) compared to the VIP immobilized for 3 days and 61% (p<0.0001) compared to the control muscles. By 2–4 weeks of immobilization the subsequent remobilization did not aggravate the initial damage. We conclude that the morphology of VIP muscle immobilized for 3 days suffers more during a subsequent remobilization period than those immobilized for 14–28 days. These findings focus attention on the mechanisms operating at the onset of disuse muscle atrophy.
An experimental model for acute poststreptococcal glomerulonephritis in mice
Authors: Nordstrand Annika , Norgren Mari , Holm Stig E.
Published: August 2009

A number of factors have been implicated in the pathogenesis of acute poststreptococcal glomerulonephritis (APSGN). The lack of a reliable animal model has made it difficult to further examine the role of these factors in the pathogenetic process. In this report, we present a tissue cage model in mice for the study of APSGN. Morphological and immunohistological changes in the kidney, resembling those of APSGN in man, were induced at high frequency in the experimental model after infection with group A streptococcal nephritis isolates. Nephritis‐associated strains induced hypercel‐lularity, occlusion of capillaries, and C3 deposition at high frequencies compared to the changes induced in animals infected with a non‐nephritis‐associated strain and non‐infected controls. In animals infected with a nephritis isolate, hematuria and proteinuria were also detected. If penicillin treatment was initiated on the third day of infection, the development of the nephritis process was prevented. Streptokinase, as well as preabsorbing antigen and streptococcal pyrogenic exotoxin B (SpeB), have been implicated in the pathogenesis of APSGN. These proteins, as well as SpeA and SpeF, were detected in the fluids of the infectious focus, regardless of the origin of the strains and whether or not glomerulonephritis was seen. Antibodies to streptokinase were evoked in the majority of the infected animals. This immune response did not correlate with the nephritic process since hypercellularity was also seen in animals which lacked detectable streptokinase antibodies. The results show that the mouse tissue cage model can be used to study APSGN and to evaluate factors involved in the pathogenesis of the disease.
The role of the Lps gene in experimental ulcerative colitis in mice
Authors: Lange Stefan , Delbro Dick S. , Jennische Eva , Mattsby‐Baltzer Inger
Published: August 2009

The effects of the Lps gene on the development of experimental ulcerative colitis were studied in two genetically different mouse strains: C57B1 and C3H. Acute colitis was induced by adding 3% dextran sulfate sodium (DSS) to the drinking water for a 7‐day (C57B1 and C3H) or a 10‐day (C57B1) experimental period. Although the DSS treatment initiated the same type of morphological changes in the colon in all groups of mice, an earlier onset and persistent intestinal bleeding occurred in the Lpsn mice (sensitive to lipopolysaccharide, LPS) in comparison with the Lpsd mice (hyporesponsive to LPS). Rectal bleeding appeared on day 7 in 90% of the Lpsn compared to 13% of the Lpsd mice (p<0.0001). In C57B1 mice, followed for three additional days, 50% of the Lpsn mice died and the surviving animals showed as well as rectal bleeding a large number of Gram‐negative bacteria in the liver and spleen. In contrast, the Lpsd mice of the C57B1 strain appeared unaffected by the treatment, although a transient rectal bleeding occurred in 90% on day 8. Also, significantly fewer Gram‐negative bacteria were found in the liver and spleen. Even though significantly increased serum endotoxin levels were seen in all DSS‐treated groups compared to controls on day 7, the serum levels of TNFoc were significantly increased only in the Lpsn mice. In DSS‐induced colitis the Lps“ genotype conferred on the mice an increased LPS susceptibility, resulting in an augmentation of the inflammatory response to Gram‐negative bacteria and their endotoxins. The results suggest that LPS‐induced host effector mechanisms significantly enhanced the intestinal bleeding, systemic inflammatory response, and mortality in mice with DSS‐induced colitis. In addition, the host defense against the invading and system‐ically spread bacteria most probably involved additional genes.
Migration of IL‐2‐activated natural killer cells in vitro: influence of extracellular matrix proteins
Authors: Unger M. L. , Johansson B. R. , Nannmark U.
Published: August 2009

An experimental set‐up for estimating a) cellular migration under agarose and b) response to chemoattractant gradients built up in the agarose was used in order to explore the behavior of adherent interleukin‐2 (IL‐2)‐activated natural killer (A‐NK) cells on cell culture plastic and after coating with extracellular matrix (ECM) constituents. A‐NK cells were deposited in wells in the agarose and directed migration, chemotaxis, towards aggregates and suspensions of B16F10 melanoma cells, suspensions of YAC‐1 cells, and tumor‐conditioned media, all deposited in wells at a 2.5 mm distance, was tested. A‐NK cell'chemotaxis was exclusively observed when B16F10 aggregates were used as attractants. The substrate influenced chemotaxis considerably, untreated plastic surface being most favorable for a chemotactic response, followed by laminin, fibronectin, and collagen IV pretreatments. Coating with reconstituted basement membrane matrix (Matrigel®) gave lesser random movements, chemokinesis, of A‐NK cells than coating with the purified components laminin and collagen IV, and the least motile response was obtained after collagen I pretreatment. These in vitro observations indicate that melanoma cell aggregates release humoral factors of a probably short‐lived nature with a chemoattractant effect on A‐NK cells, and that ECM composition influences migratory response, both conclusions with a bearing on the understanding of A‐NK cell infiltration into tumors in vivo.
Aminoglycoside resistance among Danish blood culture isolates of coagulase‐negative staphylococci
Authors: Busch‐SØRensen Charlotte , Frimodt‐MØLler Niels , Miller GEORGE H. , Espersen Frank
Published: August 2009

A sample of 137 coagulase‐negative staphylococci isolated from blood cultures in Denmark over a 4‐month period during 1992–1993 were tested for aminoglycoside resistance and for the presence of aminoglycoside‐modifying enzymes. This was done on the basis of minimum inhibitory concentrations (MICs) measured by agar dilution, inhibition zone diameter by disk diffusion, and DNA dot blot analysis. Using the National Committee for Clinical Laboratory Standards (NCCLS) MIC breakpoints, 5%, 46%, 57% and 63% of the strains were resistant to netilmicin, amikacin, gentamicin and tobramycin, respectively. The large majority of resistant staphylococci strains produced the bi‐functional AAC(6)III+APH(2“) enzyme. The presence of AAC(6‘)‐III+APH(2”) explains the high level of resistance to gentamicin, kanamycin and tobramycin. In contrast to our results, Staphylococcus haemolytkus strains are usually reported to be more resistant than Staphylococcus epidermidis strains.
Bacterial colonization and healing of venous leg ulcers
Authors: Madsen SøRen Munk , Westh Henrik , Danielsen Lis , Rosdahl Vibeke Thamdrup
Published: August 2009

The aim of the study was to evaluate a possible influence of selected bacterial species on healing of venous leg ulcers. Fifty‐nine patients with venous leg ulcers were followed via frequent semiquantitative culture of bacteria from the ulcer surface and determination of the ulcer area over a period of 180 days. Occurrences of cellulitis were treated with systemic antibiotics. There was a significant difference in relative areas on days 90 and 180 when ulcers with growth of Pseudomonas aeruginosa were compared to those without (p=0.0080 and 0.0133, respectively). Ulcers with P. aeruginosa were characterized to a great extent by enlargement in contrast to those without. Ulcers with growth of Staphylococcus aureus or haemolytic streptococci healed significantly more slowly than those without when relative areas were compared on day 180 (p=0.0079 and 0.0492, respectively). Complete healing within the observation period of 180 days was observed in 10.5% of patients with P. aeruginosa and 35% of those without (p=0.0631), in 21.6% of patients with S. aureus and 62.5% of those without (p=0.0278), and in 10.5% of patients with haemolytic streptococci and 35% of those without (p = 0.0631). The initial areas of ulcers colonized with P. aeruginosa or 5. aureus were significantly larger than those without, but no significant correlation between initial areas and ulcer healing was revealed. Conclusion: Our results suggest that P. aeruginosa in venous leg ulcers can induce ulcer enlargement and/or cause a healing delay. The results also suggest a healing delay caused by 5. aureus and haemolytic streptococci. However, conclusions have to be treated with caution since P. aeruginosa was found in combination with haemolytic streptococci in 15.3% of the patients.
Human papillomavirus infection in progressive and non‐progressive cervical intraepithelial neoplasia
Authors: Bertelsen BjøRn , Kalvenes May Britt , Hartveit Flora
Published: August 2009

Human papillomavirus (HPV) infection is common in cervical intraepithelial neoplasia (CIN) and is widely held to be responsible for its progression to grade 3. This thesis is examined here. Comparison of the level of HPV changes in 133 lesions that had not progressed to that in those from 197 women with histologically proven CIN 3 failed to reveal significant differences in their level of HPV infection on cytology, histology or in situ hybridization. However, in both these groups, some of the cases that did not show HPV positivity on in situ hybridization with probes reacting with the common HPV types did show evidence of HPV DNA using a general primer‐mediated polymerase chain reaction. This may indicate low‐copy number infections or non‐productive infections. Such reactions were more frequent in the women with progressive lesions, and it is probable that they may also have been at greater risk of cervical infection in general. The present findings suggest that a further factor, a co‐carcinogen, may be involved in progression to CIN 3, HPV being a common forerunner, providing a proliferative environment and thus favoring such an event.
Serum antibodies against Bovrelia afzelii, Borrelia burgdorferi sensu stricto and the 41‐kiloDalton flagellin in patients from a Lyme borreliosis endemic area: analysis by EIA and immunoblot
Authors: Nilsson Ingrid , Von Rosen Inger Andersson
Published: August 2009

To evaluate the possible importance of antigenic heterogeneity in the serological diagnosis of Lyme borreliosis a study was performed using antigens from various Lyme Borrelia strains. Serum samples from 102 patients with clinical signs of the infection, all living in an endemic area in southern Sweden, were evaluated by four enzyme immuno assays (EIA). The sera were initially tested for the immunoglobulin G response to antigens from a local Borrelia afzelii strain (ACAI). Serum samples from healthy blood donors residing in the same region were used to define seropositivity in the ACAI‐EIA. Immunoblotting was performed with the ACAI antigen and the reactive bands were analysed. A serum was defined as positive when at least four of the Borrelia specific polypeptides (OspC., OspA, OspB, p39, p41 [flagellin], p83, p94, 110kDa) were stained. The same sera were then analysed in three other IgG enzyme immunoassays, one based on antigens from Borrelia burgdorferi sensu stricto B31, and another on pooled protein fractions from strains B31 and ACAI. In the third EIA, sera were analysed for antiflagellin reactivity (B. afzelii strain DK‐1). An inconstant immune response was demonstrated in the EIAs and the seropositivity varied between 30–47% when low positive values were excluded, and between 38–73% if all values were included. Fifty sera (50/102) met the criteria for a positive immunoblot, but positive immunoblots were detected with both low positive and negative sera independent of antigen used in the EIAs. Antigens of the local B. afzelii strain were found to detect a higher number of seropositive individuals, which suggests that the antibody reactivity to Lyme Borrelia increases when antigens from a strain endemic in a particular geographical region are used. Data from this study suggest that EIA alone seems insufficient for the serodiagnosis, and antigenic heterogeneity of Lyme Borrelia spp. influences the performance of serum antibody tests. The reliability of serological assays could be increased when the serum antibodies against antigens of Borrelia spp. predominant in the local geographical region are measured.
Pathology of the heart after overhydration with glycine solution in the mouse
Authors: Hahn Robert G. , Zhang Weiben , Rajs Jovan
Published: August 2009

Glycine solution, which is widely used for irrigation in endoscopic surgery, has recently been suspected to have acute or subacute cardiotoxic properties. Therefore, the hearts of 46 mice that survived an intravenous infusion of 0.2 1/kg glycine 1.1%, 1.5% or 2.2% were examined for pathological alterations between 2 h and 7 weeks after the infusions. Half the animals developed extensive dilatation of the interstitium. Hypoxic events were evidenced by leakage of myoglobin from myocytes in 32 animals and correlated with dilatation of the interstitium. Focal necrosis with an inflammatory cell reaction and replacement of the myocardial tissue with granulation and fibrous tissue were observed after five glycine infusions. Pathological changes were mild or absent in five mice given mannitol solution and in three to which no infusion was given.
Fall‐off in immunity following diphtheria revaccination ‐ an 8 year follow‐up study
Authors: Simonsen Ole , Kristiansen Max , Aggerbeck Henrik , Hau Christian , Heron Iver
Published: August 2009

Diphtheria may occur even among previously vaccinated persons and knowledge of the duration of immunity is of crucial importance when designing effective vaccination programmes. In a follow‐up study of 42 representative probands revaccinated 8 years previously, a continuous fall‐off in antitoxic immunity was demonstrated. 98% were still protected (antitoxin concentration >0.01 IU/ml). From the distribution of titres in the group the individual risk of susceptibility 8 years after revaccination was calculated to be 0.8%00 (0.2–2.9%00, 95% confidence limits). Thus, repeated revaccinations are required to secure continuous protection. The fall‐off pattern for diphtheria antitoxin was approximately the same as for tetanus antitoxin. Peak values following revaccination are decisive for the duration of immunity. As peak values following vaccination depend on naturally acquired immunity and consequently decrease as indigenous diphtheria in a population disappears, highly potent vaccines are required to secure long‐term immunity following diphtheria revaccination. The effects of dose and adjuvant are discussed.
Sulphatide and its precursor galactosylceramide influence the production of cytokines in human mononuclear cells
Authors: Buschard Karsten , Diamant Marcus , Bovin Lone F. , MÅNsson Jan‐Eric , Fredman Ram , Bendtzen Klaus
Published: August 2009

Sulphatide is expressed in the central and peripheral neural system, in islets of Langerhans, and in tissues affected by late diabetic complications. Autoantibodies to sulphatide are present in patients with insulin‐dependent diabetes and the Guillain‐Barré syndrome. Cytokines influence these disease processes, and we therefore studied whether sulphatide and its precursor galactosylceramide (gal‐cer) influence the in vitro production of cytokines by blood mononuclear cells (MNC) originating from 15 healthy persons. Using lipopolysaccharide (LPS)‐stimulated cells, sulphatide increased the IL‐2 production (163±17% of controls without sulphatide, p=0.02), and gal‐cer increased the IL‐1α production (145±13%, p=0.006), whereas neither gal‐cer nor sulphatide had an effect on the production of IL‐6, IL‐10 or TNFα. When stimulating cells with phytohaemagglutinin (PHA), sulphatide decreased the production of IL‐6 (88±5%, p=0.009), IL‐10 (66±3%, p=0.000003), and TNFα (75±9%, p=0.02). Galcer, however, increased the production of IL‐6 (188±13%, p=0.000006), and decreased the production of TNFp (80±6%, p=0.007). Neither galcer nor sulphatide had an effect on the production of IL‐2 or IFNy from PHA‐stimulated cells. Northern blot analysis using an IL‐6 probe similarly showed an increased amount of IL‐6 mRNA after galcer incubation (range 469%‐150%, n=3) of PHA‐stimulated control. Thus, sulphatide and galcer influence the production of several cytokines thought to be involved in immunoinflammatory disease processes.
Cutaneous sparganosis in France: the second case described from Europe
Authors: GARIN Y. J. F. , FROTTIER J. , LAVERGNE‐SLOVE A. , HOUDART R. , POIROT J. L.
Published: August 2009

The first case of sparganosis is reported from France. The patient, a 21‐year‐old man, presented with a subcutaneous lump on the chest, and the diagnosis was made on histological examination after needle biopsy. He achieved a complete recovery.
Microvascular degeneration in hereditary cystatin C amyloid angiopathy of the brain
Authors: WANG ZHEN ZHEN , JENSSON OLAFUR , THORSTEINSSON LEIFUR , VINTERS HARRY V.
Published: August 2009

Hereditary cystatin C amyloid angiopathy (HCCAA), an autosomal dominant form of cerebral amyloid angiopathy (CAA) occurring primarily in Iceland, is characterized by a variant cystatin C amyloid deposition in the walls of cerebral parenchymal and leptomeningeal vessels. Cystatin C is also found to colocalize with amyloid ß/A4 protein in cerebral vessel walls of patients with Alzheimer's disease (AD), sporadic CAA, and hereditary cerebral hemorrhage with amyloidosis, Dutch type (HCHWA‐D). The abundance of cystatin C deposition in cerebral blood vessel walls suggests that cellular elements of the vessel wall itself may play a role in its deposition. Microvascular changes in the brains of HCCAA patients were investigated by single‐ and double‐label immunohistochemistry. We found that cystatin C amyloid immunoreactivity was present not only in cerebral cortical and leptomeningeal vessels, but also in white matter parenchymal vessels. Cystatin C deposition was more prominent in the media of parenchymal vessels and in the adventitia of leptomeningeal vessels. Smooth muscle (sm) cells were few or could not be identified within vessel walls showing extensive cystatin C deposition, suggesting progressive loss of these cells as cystatin C accumulates. However, in less severely affected vessels, cystatin C was present in cells that also had the phenotype of sm, suggesting that sm cells synthesize or process cystatin C. Cystatin C immunoreactivity was in addition, detected in some neuronal cell bodies throughout the cortex in patients with HCCAA and AD‐related CAA. Our results indicate that cellular components of the vessel walls may play an important role in cystatin C deposition, as they do in β/A4 deposition in AD‐related CAA. Cystatin C deposition within the vascular media and adventitia, with associated vessel wall injury as manifested by sm cell loss, represents microvascular degeneration that leads to cerebral hemorrhage.
Interaction between proximal tubular cells and allogenic leukocytes mediated by interleukin‐1 and prostaglandin
Authors: BLAEHR HANNE , ANDERSEN CLAUS B. , LADEFOGED JOERGEN
Published: August 2009

The present study investigates the interaction between proximal tubular cells (TC) and peripheral blood mononuclear cells (PBMC) with respect to interleukin‐1 (Il‐1) and prostaglandin E2 (PGE2) in co‐culture experiments. Stimulator cells were proliferating human TC propagated in culture and characterized by expression of ICAM‐1 and MHC‐class II in response to cytokines. The presence of TC reduced spontaneous growth of PBMC by 42%, with a concomittant 50% reduction of Il‐1 release and a 6‐fold increase of PGE2 release in the growth medium. The growth reduction was partly counteracted by addition of Il‐1 and indomethacin. Addition of Il‐1 stimulated the PGE2 release from PBMC and TC by about 75% and 500%, respectively. Growth of TC cultured alone was inhibited by Il‐1 in a dose‐dependent manner, with maximum effect at 10 U/ml, whereas PGE2 showed no effect. These results suggest that TC have the capacity to interact with PBMC, and this potentially tissue‐protective mechanism induced by TC may have clinical implications.
Correlation between polymerase chain reaction and cervical cytology for detection of human papillomavirus infection in women with and without dysplasia
Authors: GJØEN K. , SAUER T. , OLSEN A. O. , ØRSTAVIK I.
Published: August 2009

The aim of this study was to compare the ability of two methods, the polymerase chain reaction (PCR) and cervical cytology, to detect HPV infection. The study population included 222 randomly selected women without dysplasia (controls) and 91 women with histologically confirmed dysplasia (CIN II‐III) (cases). In women without dysplasia, 8.6% had cytological signs of HPV infection, whereas 15.3% were HPV DNA positive by PCR. In women with dysplasia, 72.5% had cytological signs of HPV infection, whereas 90.1% were HPV PCR positive. The statistical agreement between the two diagnostic methods was low (controls: k= 0.26, cases: k= ‐0.03). In total, PCR failed to detect 17 of 85 women with cytological signs of HPV infection, whereas cervical cytology failed to detect 48 of 116 HPV PCR‐positive women. In women with dysplasia, but not in women without dysplasia, the oncogenic HPV types were associated with cytological signs of HPV infection.
Postmortem blood cultures
Authors: SCHØNHEYDER HENRIK C. , HENRIQUES ULRIK
Published: August 2009

Postmortem blood cultures were obtained by separate puncture of the right and left side of the heart during autopsy of 18 patients with solid cancer and 26 patients with non‐neoplastic diseases. Concordant isolates were obtained from the right and left side in six patients (14%, 95% confidence limits: 5–28%). Growth was unilateral in 7 patients and cardiac aspirates were sterile in 31 patients (70%, 95% CL: 55–83%). One patient had been treated for Staphylococcus aureus bacteraemia and eight patients had negative premortem blood cultures. Separate culture of blood from the right and left side of the heart may add to the information gained by postmortem blood culture.
Antigenic peptides and autoimmunity
Authors: SINIGAGLIA FRANCESCO , STURNIOLO TIZIANA , RADDRIZZANI LAURA , HAMMER JUERGEN
Published: August 2009

MHC class II alleles play a major role in determining resistance or susceptibility to autoimmune disease. Considerable effort is being expended to establish the role polymorphisms play in influencing the binding of antigens, including autoantigens, in the peptide‐binding groove. Single amino acid substitutions in the MHC cleft, for instance at DRβ71 and DQβ57, influence peptide binding. Although candidate autoantigenic peptides have been identified which bind to disease‐associated MHC molecules, several critical questions remain to be answered before the role of these peptides in the autoimmune disease process can be established.
Ultrastructure of colony‐like communities of bacteria
Authors: TETZ VICTOR V. , RYBALCHENKO OKSANA V.
Published: August 2009

Colony‐like communities are poorly studied forms of bacterial growth on agar. These communities are formed after the growth of large amounts of bacteria simultaneously plated onto a limited area of agar, while “classical” colonies are formed as a result of single bacterial cell multiplication. Colony‐like communities of Gram‐negative and Gram‐positive bacteria differ from “classical” microbial colonies in their ultrastructural organization. Almost every cell in colony‐like communities has an individual capsule‐like envelope (glycocalyx). The cells in these communities are characterized by accelerated ageing. In the course of their development both bacterial colony‐like communities and “classical” colonies produce a film, the basic part of which is represented by an elementary membrane. In contrast to “classical” colonies, the thickness of the amorphous layers of this film in colony‐like communities did not significantly increase after 24 h of cultivation. The formation of a three‐dimensional network of cells in colony‐like communities is similar to this process in “classical” colonies. The intercellular matrix of colony‐like communities contains numerous membrane vesicles, and has a more amorphous structure and higher electron density than that of “classical” bacterial colonies.
Comparison of light microscopy, in situ hybridization and polymerase chain reaction for detection of human papillomavirus in histological tissue of cervical intraepithelial neoplasia
Authors: LIE A. K. , SKJELDESTAD F. E. , HAGEN B. , JOHANNESSEN E. , SKARSVÅG S. , HAUGEN O. A.
Published: August 2009

The aims of the study were to estimate the prevalence of HPV infection in patients treated for high grade lesions of the cervix uteri (HG CIN), and to evaluate the validity of the histological criteria used for detection of HPV infection. The study comprised 203 women treated for HG CIN by laser conization. Forty‐three preoperative biopsies and 160 cone specimens were examined for HPV infection using light microscopy (LM), in situ hybridization (ISH), and polymerase chain reaction (PCR). ISH was performed using commercial biotinylated probes for HPV types 6/11, 16/18, and 31/33/35 (Vira‐Type In Situ Kits, Digene Diagnostics, Silver Spring, MD). HPV‐PCR was performed with the L1 consensus primers Gp5+/6+. The prevalence of HPV detected by LM was 70%; by ISH 48% and by PCR 83%. Using PCR as the gold standard, LM had a sensitivity of 0.71 and specificity of 0.41. The corresponding results for ISH were 0.51 and 0.65, respectively. The positive predictive value for both tests reached over 80%, but the negative predictive value was less than 25%. This study demonstrates that morphology is an unspecific method of identifying HPV infections. LM identification of HPV infections has no clinical implications. Our analyses comparing test performances of LM, ISH and PCR show that PCR is the superior method.
Epidemiological study of Acinetobacter species isolated from an intensive care unit
Authors: GARCÍA‐ARATA M. ISABEL , GERNER‐SMIDT PETER , LÓPEZ‐BREA MANUEL
Published: August 2009

An epidemiological survey of the increase in Acinetobacter species isolates occurring in the intensive care unit of a Spanish teaching hospital during 1993 and 1994 was carried out. Different laboratory methods were used to find out, whether there was a genetic linkage. The isolates were divided into three main groups according to the resistance patterns to 11 drugs. Using API 20NE biotyping, eight different types were found. The two most common contained 20 and 11 isolates, respectively. Five different plasmid profile types were observed, although plasmids were only demonstrated in 40% of the isolates. Ribotyping with EcoRI, Sa/I and ClaI enzymes revealed 10, 9, and 8 different patterns, respectively. In total, 15 different ribotypes were identified using these three enzymes. Twenty‐one isolates belonged to exactly the same ribotype, and 13 were associated with two highly related ribotypes. In the first ribotype, only five isolates harboured plasmids. The ribotyping method produced 100% typability and ribotypes were easy to compare; it also had taxonomic value. Ribotyping allowed us to determine the genetic linkage between Acinetobacter isolates recovered from ICU patients.
Immunoelectron microscopy on epoxy sections without deplasticizing to detect glomerular immunoglobulin and complement deposits in renal diseases
Authors: BRORSON SVERRE‐HENNING , STRØM ERIK HEYERDAHL , SKJØRTEN FREDRIK
Published: August 2009

Twenty renal biopsies were studied by immunoelectron microscopy (IEM) after embedding in epoxy resin. Immunogold labeling for immunoglobulins and complement C3 was performed on the epoxy sections, which were not subjected to any kind of etching or deplasticizing prior to the immunolabeling. The concentration of accelerator, DMP‐30 (Tri (Dimethyl Amino Methyl) Phenol), was increased in the infiltration and embedding steps far beyond the values normally used to make immunolabeling of these antigens possible on epoxy sections. The sections were stained with tannic acid accompanied by uranyl acetate and lead citrate. Immunofluorescence (IF) for light microscopy was carried out on frozen sections of parallel tissue samples. Some cases with IgA‐nephritis demonstrated a higher sensitivity for IEM than IF, in the sense that smaller amounts of antigen were detectable with IEM. Ultrastructural preservation with this method was approximately the same as that usually seen on epoxy‐embedded material. By combining excellent immunolabeling with nearly optimal ultrastructural morphology in one procedure, this method is useful particularly in situations where the material available is limited, such as in studies of renal biopsies. As far as we know, this is the first time that immunoglobulins have been satisfactorily immunolabeled on epoxy sections without etching or deplasticizing.
Production and characterization of mouse monoclonal antibodies against Afipia felis
Authors: ENGBÆK KRÆSTEN , UTTENTHAL LARS OTTO , KOCH CLAUS
Published: August 2009

A series of 10 monoclonal antibodies reacting with Afipia felis antigens were selected from mice immunized with live organisms of the reference strain ATCC 53690. Immunoblotting against SDS‐PAGE‐separated A. felis sonicate allowed the antibodies to be classified into three groups: 1) 168‐4, ‐6, ‐7 and ‐10 reacted with a 53 kDa antigen, 2) 168‐1, ‐3 and ‐9 reacted with both 53 kDa and 60 kDa antigens, and 3) 168‐2, ‐5 and ‐9 reacted with other antigens. Antibodies of group 1 did not cross‐react with other Afipia species or 36 unrelated bacteria, whereas those of groups 2 and 3 reacted with other Afipia species and some unrelated bacteria. Immunoblots of crossed immunoelectrophoretic patterns of A. felis sonicate against rabbit antiserum showed that antibodies of groups 1 and 2 bound to the same precipitin arcs. Antibodies of group 1 reacted with a species‐specific epitope on the 53 kDa antigen, while those of group 2 reacted with other epitopes shared by the 53 kDa and 60 kDa antigens. The binding of antibodies of group 1 to A. felis sonicate was inhibited by post‐infection rabbit serum, whereas no inhibition was observed for antibodies of group 2. The species‐specific epitope of the 53 kDa antigen and the early appearance of antibodies against this epitope after infection suggest that this antigen can be used in a serodiagnostic test for A. felis infection.
HPV 16 infection in a patient with two primary squamous cell carcinomas: of the uterine cervix and the anal mucosa
Authors: BERNER AASMUND , FRANZÉN SIXTEN , HOLM RUTH
Published: August 2009

A 43‐year‐old woman presented with metrorrhagia and a polypoid anal tumor. Cervical dilatation and endocervical curettage and resection of the anal tumor revealed two primary squamous cell carcinomas: of the uterine cervix and the anal mucosa. HPV 16 was identified in both tumors using an in situ hybridization technique with probes against HPV 6, 11, 16, 18, 31 and 33. The cervical carcinoma was weakly positive for p53 protein, whereas the anal tumor was p53 protein negative. Five years later a recurrence of the cervical carcinoma was diagnosed and a hysterectomy was performed. Of 3,013 patients with squamous cell carcinoma of either the cervix or the anus registered in our files, only one had a primary carcinoma in both locations. It was concluded hat HPV 16‐induced squamous cell carcinoma may occur simultaneously in the cervix and the anus, and carcinoma in either of the two locations has to be ruled out when dealing with HPV infection in the anogenital tract.
Human immunoglobulin isotype and IgG subclass response to different antigens of Moraxella catarrhalis
Authors: RAHMAN M. , HOLME T. , JÖNSSON I. , KROOK A.
Published: August 2009

Enzyme immunoassays were tested for the determination of antibodies of different isotypes and IgG subclasses to Moraxella catarrhalis in human sera. An assay based on an outer membrane protein preparation (OMP) as antigen was compared to assays using whole bacterial cells and a purified lipopolysaccharide preparation. There was a good correlation between the results obtained with the OMP preparation and the whole‐cell antigen. In paired sera, optimal sensitivity was obtained by using the OMP preparation as coating antigen and testing for a rise in IgG3 antibodies. However, patients with high levels of antibodies in acute serum had no or only an insignificant antibody response during infection.