bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunopurified extracellular Bartonella henselae antigen for detecting specific antibodies by enzyme immunoassay
Authors: ENGBÆK KRÆSTEN , UTTENTHAL LARS OTTO , KOCH CLAUS
Published: August 2009

Protein antigens of Bartonella henselae bacterial sonicate supernatant and concentrated cell‐free culture filtrate were examined by SDS‐PAGE. The sonicate supernatant gave 38 bands and the culture filtrate at least 21, of which 18 were of bacterial origin. Immunoblotting against 13 monoclonal antibodies obtained from mice infected with live B. henselae showed that 10 of these antibodies reacted with a narrow 225 kDa band and varying smears of bands ranging from 36 to 240 kDa in the sonicate, but only with a single 200 kDa band in the culture filtrate. Testing of pre‐ and post‐infection rabbit sera in immunoblotting against culture filtrate demonstrated that the 200 kDa component gave the most prominent specific reaction with post‐infection sera. The 200 kDa antigen was isolated by immunoaffinity chromatography of concentrated culture filtrate, and its molecular size determined by size‐exclusion chromatography as >1000 kDa. The immunopurified antigen was compared with bacterial sonicate as coating antigen in EIA for determining humoral immune responses in rabbits inoculated with live B. henselae. The two antigens gave almost identical results for IgM and IgG responses. The specificity of the immunopurified antigen was tested in EIA against hyperimmune rabbit sera and sera of rabbits inoculated with live B. henselae, B. quintana and Afipia felis. Only the hyperimmune serum against B. henselae and the sera of the rabbits inoculated with live B. henselae reacted with the immunopurified antigen, whereas the B. henselae sonicate cross‐reacted with hyperimmune and post‐infection sera of rabbits inoculated with B. quintana and A. felis.
Rapid detection of Helicobacter pylori infection in serum and whole‐blood samples
Authors: ENROTH HELENA , ENGSTRAND LARS
Published: August 2009

The aims of this study were to evaluate the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and diagnostic accuracy of a rapid serum and whole‐blood test, Quick Vue One‐Step H. pylori test (Quidel Corporation)) for detection of Helicobacter pylori infection. We tested 79 patients with gastrointestinal symptoms, including 8 gastric cancer patients. In total, 116 rapid tests were performed: both serum and whole‐blood samples were tested in 37 patients (74 tests) and serum only was tested in a further 42 patients (42 tests). Serology by ELISA, bacterial culture and histology were also carried out on samples from these patients and used as standards for determining H. pylori infection. Diagnostic accuracy of the tests was 0.76 for both sera and whole‐blood when culture and histology was used as the standard in the calculations. However, using serology by ELISA as the standard raised the diagnostic accuracy to 0.80 in serum samples and to 0.89 in whole‐blood samples. Overall, the H. pylori status observed when using the rapid tests correlated well with serology, bacterial culture and histology in this study.
Influence of aqueous extracts of medicinal plants on surface hydrophobicity of Escherichia coli strains of different origin
Authors: TÜURI MALL , TÜURI ENDEL , KÕLJALG SIIRI , MIKELSAAR MARIKA
Published: August 2009

The adhesion of microbes on host cells is of decisive importance in the development of Gram‐negative microbe‐induced infections and can be influenced by the surface hydrophobicity of the microbial cell. The hydrophobicity of 155 Escherichia coli strains of different origin was determined by the salt aggregation test (SAT). Among the strains isolated from faecal samples of healthy persons only 16.7% showed aggregative properties, whereas among the strains isolated from the urine of patients with pyelonephritis and the faecal samples of calves and pigs with diarrhoea some 40.0%‐60.0% were aggregative. The influence of aqueous extracts prepared from bearberry leaves, St. John's wort herbs, wild camomile and marigold flowers on hydrophobicity of 40 E. coli and 20 Acinetobacter baumannii strains was investigated. The decoctions of bearberry and St. John's wort increased remarkably the hydrophobicity of both microbial species. The infusions of wild camomile and marigold completely blocked the aggregative properties of the investigated strains. Bactericidal action was relatively low in the case of bearberry and St. John's wort and completely lacking in the case of wild camomile and marigold. Thus, one of the probable and potentially important action mechanisms of the four medicinal plants studied is their ability to influence the surface characteristics of the microbial cells and thereby their putative virulence properties.
Prognosis of vulvar dysplasia and carcinoma in situ with special reference to histology and types of human papillomavirus (HPV)
Authors: JUNGE JETTE , POULSEN HEMMING , HORN THOMAS , HØORDING ULLA , LUNDVALL FINN
Published: August 2009

Sixty‐one women with vulvar dysplasia or carcinoma in situ were treated with local laser excision of the initial lesion and of the recurrences, and followed at intervals of from 3 increasing to 12 months. Recurrences were observed in 16 (26%) patients. No case of invasive carcinoma was seen. Patients with recurrences were significantly younger than those without (p<0.02, median age 42.5 and 54 years, respectively). The resection borders were significantly more often involved in the initial lesions in the group with recurrences (36%) than in the group without (9%) (p<0.014). All lesions were classified according to the WHO (mild, moderate, severe dysplasia or carcinoma in situ) and Toki et al. (1991) (warty, basaloid, combined warty/basaloid or mixed (warty, basaloid and simple). No pure types of Toki (1991) could be demonstrated. There were no differences regarding recurrences in any of these groups. HPV DNA was detected in the initial lesions by PCR in 50/56 (89%) (44 with HPV type 16 and 6 with HPV type 33) and by ISH in 23/61 (38%). The same type of HPV could be demonstrated in all first recurrences except in two, where HPV type 33 was shown in specimens harboring HPV type 16 in the initial lesions. In one of these cases, HPV type 16 could again be demonstrated in the second and final recurrence. In no specimen was more than one type of HPV detected. The results indicate that the most important parameter in predicting the recurrence of vulvar dysplasia or carcinoma in situ is the involvement of the resection borders. The location of the lesion, the degree and type of dysplasia, and the type of HPV seem to play a minor role. Local excision and subsequent intensive control with removal of any visible new lesion probably prevents development of vulvar invasive carcinoma.
Viridans streptococci in blood cultures. Can we see any patterns of species related to patient category?
Authors: DWYER R. , RINGERTZ S.
Published: August 2009

We present a review of 229 blood cultures with viridans streptococci collected during a period of eight and a half years from 1986 to 1994 at a teaching hospital in Sweden. The clinical significance of the growth of viridans streptococci is always uncertain, since these bacteria can be contaminants from the skin flora. Growth in more than one culture bottle strengthens the clinical value of the finding. The question was whether species identification might also help in the assessment of the clinical relevance of the finding. The results show that Streptococcus mitis occurs significantly more frequently in blood cultures from the departments dealing with cancer patients. Even with polymicrobial growth in blood cultures, S. mitis should be considered a pathogen of clinical relevance, not a contaminant.
Melanoma‐associated antigens recognized by cytotoxic T lymphocytes
Authors: KIRKIN ALEXEI F. , DZHANDZHUGAZYAN KARINE , ZEUTHEN JESPER
Published: August 2009

During the last 7 years significant progress has been made in the identification of melanoma‐associated antigens recognized by cytotoxic T lymphocytes (CTL). These antigens belong to three main groups: cancer/testis‐specific antigens (MAGE, BAGE, GAGE, PRAME and NY‐ESO‐1), melanocyte differentiation antigens (tyrosinase, Melan‐A/MART‐1, gp 100, TRP‐1 and TRP‐2), and mutated or aberrantly expressed antigens (MUM‐1, CDK 4, β‐catenin, gp 100‐in4, p15 and N‐acetylglucosaminyl‐transferase V). In this review we have summarized the available data concerning the characterization of melanoma‐associated antigens, focusing on their immunogenic and protective properties. The development of a strong immune response to differentiation antigens is limited by the existence of tolerance to these “self” ‐antigens, permitting the involvement of only T cells with low affinity T‐cell receptors. Among the melanoma differentiation antigens, only gp 100 has been shown to be a tumor regression antigen. The cancer/testis‐specific antigens such as MAGE and PRAME should potentially be highly immunogenic antigens. They contain several potential HLA class I binding epitopes and are present only in the testes, which are not accessible to the cells of the immune system owing to the lack of direct contact with the immune cells and the lack of HLA class I expression on the surface of germ cells. But only two patients have been found who responded to these antigens in vivo, indicating their genuinely low immunogenicity. A comparison of the predicted secondary structures of these two groups of antigens (cancer/testis‐specific and differentiation antigens) revealed enrichment of long α‐helical stretches in the cancer/testis‐specific antigens. We hypothesize that such highly organized stable structures could, first, reduce denaturation of the protein and, thus, ubiquitinylation as a degradation signal, and, second, diminish the efficiency of the protein unfolding – a necessary step in the proteolytic cleavage by proteasomes. High structural stability could therefore be responsible for the low immunogenicity of these proteins. In this case, modifications decreasing the stability of these proteins might be a means of improving the immune response to these potentially therapeutically useful antigens.
Diagnosis of congenital Toxoplasma gondii infection by polymerase chain reaction (PCR) on amniotic fluid samples
Authors: JENUM P. A. , HOLBERG‐PETERSEN M. , MELBY K. K. , STRAY‐PEDERSEN B.
Published: August 2009

As part of a screening project for detection of Toxoplasma gondii infection among pregnant women in Norway, nested polymerase chain reaction (PCR) aimed at the detection of T. gondii in amniotic fluid samples was included in the diagnostic routine. The results were compared with the routine criteria for congenital infection: i) T. gondii detected in amniotic fluid or cord blood by mouse inoculation, ii) specific IgM or IgA in serum collected after birth, and/or iii) specific IgG persisting beyond one year of age. The PCR was based on the B1 gene with an internal control gene amplified together with the B1 gene. One hundred and two amniotic fluid samples collected during pregnancy and/or at delivery from 67 pregnant women with serological evidence of primary T. gondii infection were available for examination by both B1‐PCR and mouse inoculation. Six samples were positive and 86 samples were negative by both methods (90% concordance). One sample was mouse inoculation positive and B1‐PCR negative while nine samples were B1‐PCR positive and mouse inoculation negative, of which five were associated with four infants without proven infection. 59% and 41% of samples associated with infected infants were positive by B1‐PCR and mouse inoculation, respectively. The difference was mainly due to a lower detection rate by mouse inoculation after antiparasitic treatment. The specificity of B1‐PCR was 94%. Even though B1‐PCR performed on amniotic fluid samples did not detect all infected infants, it represented a valuable tool in addition to conventional methods in the diagnosis of congenital T. gondii infection.
Pulmonary intravascular macrophages appear in rats after long‐term administration of lipid emulsion and amino acid solution
Authors: AKSNES JON , EIDE TOR J. , NORDSTRAND KENNETH
Published: August 2009

Intravascular macrophages have rarely been seen in normal lungs of humans and rats, but in rats endotoxaemia has induced their presence. To study whether substrates used for parenteral nutrition could have a similar stimulatory effect on mononuclear phagocytes, rats were given lipid emulsion (n=5), amino acid solution (n=5), or isotonic saline (n=5) through central venous catheters for 3 weeks. Structural changes in the lung microvessels were evaluated using electron microscopy. The areal fraction of pulmonary intravascular mononuclear phagocytes was 19.6% (SD=8.2) in rats given lipid emulsion (p<0.05) and 8.2% (SD=8.2) in rats given amino acid solution n.s. compared to 2.4% (SD= 4.0) in rats given saline. The increase in areal fraction was mainly due to an increase in cell numbers. In rats given lipid emulsion the intravascular phagocytes were only slightly larger than in rats given saline, but had the morphological features of mature macrophages. The study demonstrates that lipid emulsion recruits pulmonary intravascular macrophages in rats, indicating a stimulatory effect on the mononuclear phagocyte system. The effect was less pronounced with amino acid solution.
The value of autopsy from a clinical point of view
Authors: MIDELFART JANA , AASE STEINAR
Published: August 2009

An enquiry into clinicians' attitudes to autopsies was conducted by questionnaire sent to 166 general practitioners in the county of Sør‐Trøndelag in Norway and to 186 clinicians working at the University Hospital of Trondheim. It was considered especially important for us to include general practitioners. As the result of a government decision taken at the end of 1994, autopsies requested by general practitioners on patients dying outside hospitals have from January 1995 been covered by national health insurance. Answers were obtained from 250 doctors: 110 general practitioners and 140 hospital physicians. One hundred and seventy‐nine (73.1%, n=245) felt that the possibility of having autopsies performed was of great importance in their daily work. Autopsy was considered to be a very important means of quality assurance in the health care system by 158 (66.4%, n=238). One hundred and two (41.2%, n=247) answered that improvements in medicine and technology during the last decades had not reduced the importance of autopsy. One hundred and twenty‐two (81.3%, n=150) felt that especially computer tomography (CT) had reduced the value of autopsy. Among the general practitioners, 73 (68.9%, n=106) welcomed the opportunity to have non‐forensic autopsies performed on patients who died outside hospitals. Our study showed differences in the attitudes of clinicians towards autopsies, but our results still indicate that the value of autopsy for furthering clinical knowledge is acknowledged by most clinicians.
Diagnosis of pulmonary tuberculosis
Authors: THOMSEN VIBEKE ØSTERGAARD
Published: August 2009

The purpose of this investigation was to evaluate the Amplified Mycobacterium tuberculosis Direct Test (AMTDT) for the diagnosis of pulmonary tuberculosis (TB). Sixhundred and forty‐six sputum samples were analysed by microscopy for acid‐fast bacilli, by culture for mycobacterial growth, and by AMTDT for the presence of M. tuberculosis complex rRNA. If there were discrepant results, information as to whether the patient had a history of TB was obtained. The sensitivity was 85.7% (smear positive 100% and smear negative 71.4%, respectively) and the specificity was 96.8% compared with culture. By retesting, 12 of 13 false‐positive samples could be divided into two major categories. The samples in one category were retest positive, had highly positive results, and the patients had previous culture‐proven TB. The samples in the other category had less positive AMTDT results, were retest negative, and the patients were not notified as having TB. This investigation shows that AMTDT is a sensitive supplementary method for rapid detection of M. tuberculosis complex in respiratory specimens, that there is a risk of contamination which has to be monitored closely, and that positive samples should be retested.
S‐100 protein‐positive dendritic cells in follicular lesions of the thyroid
Authors: HIROKAWA MITSUYOSHI , SHIMIZU MICHIO , MANABE TOSHIAKI , SONOO HIROSHI , HARADA TANEKAZU
Published: August 2009

Little is known about the role of S‐100 protein‐positive dendritic cells in follicular lesions of the thyroid. In 36 cases of adenomatous goitre, 18 cases of follicular adenoma, and 7 cases of follicular carcinoma, we investigated the incidence and distribution of S‐100 protein‐positive dendritic cells using immunohistochemical staining. Four cases (10.5%) of adenomatous goitre, five cases (31.3%) of follicular adenoma, and 6 cases (85.7%) of follicular carcinoma contained S‐100 protein‐positive dendritic cells in the subcapsular area and/or the hyalinized stroma. Dendritic cells in the subcapsular area were observed only in lesions with thick capsules and not those with thin capsules. In most cases of follicular carcinoma, ++ to +++ of the dendritic cells was seen together with a few lymphocytes and fibroblasts in the subcapsular areas. In contrast, cases of adenomatous goitre and follicular adenoma showed only a few dendritic cells without any inflammatory cells. Dendritic cells in follicular thyroid lesions may play some role in capsular formation or may be a secondary phenomenon due to capsular formation.
Failure to detect Chlamydia pneumoniae in calcific and degenerative arteriosclerotic aortic valves excised during open heart surgery
Authors: ANDREASEN JAN JESPER , FARHOLT STENSE , JENSEN JØRGEN SKOV
Published: August 2009

Chlamydia pneumoniae has been associated with atherosclerosis, although no causal association has been established. Employing culture and polymerase chain reaction in aortic valves with calcific and degenerative arteriosclerotic changes from 23 non‐consecutive patients undergoing aortic valve replacement, C. pneumoniae was not detected in any of the valves. 19/22 patients had serological evidence of past infection with C. pneumoniae. Our findings do not provide supportive evidence for the hypothesis that C. pneumoniae is associated with calcific or degenerative arteriosclerotic aortic heart valve disease.
Ulcer bed infection
Authors: DANIELSEN LIS , BALSLEV EVA , DÖRING GERD , HØIBY NIELS , MADSEN SØREN MUNK , ÅGREN MAGNUS , THOMSEN HENRIK KLEM , FOS HENRIK H. STEEN , WESTH HENRIK
Published: August 2009

We report a case of ulcer bed infection in an enlarging venous leg ulcer without clinical signs of cellulitis in the surrounding tissues. Signs of infection in the leg ulcer were: 1) cocci‐like structures and bacteria‐like rods around vessel walls in the viable ulcer bed, 2) vasculitis‐like inflammation of deeply situated vessels of the viable tissue, 3) Pseudomonas aeruginosa‐specific antibodies in the serum (other than against exotoxin A), 4) extensive epidermolysis of normal human skin by the wound exudate in vitro, and 5) P. aeruginosa exotoxin A in the wound exudate (23 ng/ml). In an in vitro cell assay, the wound exudate was cytotoxic and rabbit antibodies to exotoxin A, but not a serine proteinase inhibitor, inhibited this cytotoxicity. P. aeruginosa exotoxin A might contribute to the pathogenesis of the ulcer enlargement. The ulcer improved after the third skin graft, probably mainly due to effective treatment with a long‐stretch compression bandage.
Reappraisal of the expression of mast cell proteases of Mongolian gerbils (Meriones unguiculatus)
Authors: KUROKAWA MOTOKI , OGATA KATSUMI , UCHIYAMA FUKUMI , KOBAYASHI TAKAHIKO , ISHIWATA KENJI , NAWA YUKIFUMI
Published: August 2009

Mast cell proteases in the tongue and jejunum of Mongolian gerbils (Meriones unguiculatus) were examined by enzyme‐histochemical methods. Both trypsin‐like (tryptase) and chymotrypsin‐like (chymase) protease activities were demonstrated in mast cells in the tongue of fresh cryosections. When frozen sections of the tongue were post‐fixed in various fixatives, those fixed in Carnoy's fluid showed strongest enzyme activities. Tryptase and chymase activities in paraffin sections of both tissues were well preserved when tissues were fixed in Carnoy's fluid at 4°C for 15 min. However, enzyme activities in both tissues, especially in the tongue, were drastically reduced by longer fixation time and higher temperature. When Carnoy‐fixed (4°C for 15 min) paraffin sections were treated with heparinase I or chondroitinase ABC before enzyme‐histochemical stainings for proteases, tryptase activities were lost entirely in the tongue and mostly in the jejunum by heparinase I digestion, and slightly in both organs by chondroitinase ABC digestion. In contrast, chymase activities at both sites were not influenced by these pretreatments. These results show that although mast cells in the tongue as well as in the jejunum of Mongolian gerbils contain both tryptase and chymase activities, their stability to fixations is variable among organs so that tissue fixation conditions are crucial for the preservation. At least some part of the stability of mast cell proteases is dependent on the proteoglycans present in mast cell granules.
Adhesion to denture acrylic surfaces and relative cell‐surface hydrophobicity of Candida parapsilosis and Candida albicans
Authors: PANAGODA G. J. , ELLEPOLA A. N. B. , SAMARANAYAKE L. P.
Published: August 2009

C. parapsilosis is an opportunistic emerging pathogen which together with C. albicans causes diseases in immunocompromised patients. Adhesion of Candida species to various surfaces is an important event in colonization and pathogenesis, and the relative cell‐surface hydrophobicity (CSH) of the organism is a contributory physical force involved. Therefore, in vitro adhesion to acrylic surfaces and relative CSH of 24 isolates of C. parapsilosis and 10 isolates of C. albicans were studied. There was no significant difference in relative adhesion of C. parapsilosis isolates and C. albicans, although the former demonstrated a tendency for increased adhesion. There was significant intra‐species variation in adhesion among isolates of C. parapsilosis (p=0.0001), but not C. albicans. In general, C. parapsilosis isolates demonstrated a two‐fold greater relative CSH than C. albicans (p=0.0003). When the relative CSH of superficial and systemic isolates of C. parapsilosis were compared, the former showed a significantly higher (49.15%) relative CSH than their systemic counterparts (p<0.01). A highly significant positive correlation between adhesion and relative CSH of C. parapsilosis (p=0.74, p<0.0001) was also noted. Taken together, these data suggest that the attributes of adhesion and relative CSH of Candida species may contribute differentially in varying disease states of the human host, such as superficial and systemic Candida infections.
Resistance to antimicrobial agents used for animal therapy in pathogenic‐, zoonotic‐ and indicator bacteria isolated from different food animals in Denmark: a baseline study for the Danish Integrated Antimicrobial Resistance Monitoring Programme (DANMAP)
Authors: AARESTRUP F. M. , BAGER F. , JENSEN N. E. , MADSEN M. , MEYLING A. , WEGENER H. C.
Published: August 2009

This study describes the establishment and first results of a continuous surveillance system of antimicrobial resistance among bacteria isolated from pigs, cattle and broilers in Denmark. The three categories of bacteria tested were: 1) indicator bacteria (Escherichia coli, Enterococcus faecalis, Enterococcus faecium), 2) zoonotic bacteria (Campylobacter coli/jejuni, Salmonella enterica, Yersinia enterocolitica), and 3) animal pathogens (E. coli, Staphylococcus aureus, coagulase‐negative staphylococci (CNS),Staphylococcus hyicus, Actinobacillus pleuropneumoniae). A total of 3304 bacterial isolates collected from October 1995 through December 1996 were tested for susceptibility to all major classes of antimicrobial agents used for therapy in Denmark. Bacterial species intrinsically resistant to an antimicrobial were not tested towards that antimicrobial. Acquired resistance to all antimicrobials was found. The occurrence of resistance varied by animal origin and bacterial species. In general, resistance was observed more frequently among isolates from pigs than from cattle and broilers. The association between the occurrence of resistance and the consumption of the antimicrobial is discussed, as is the occurrence of resistance in other countries. The results of this study show the present level of resistance to antimicrobial agents among a number of bacterial species isolated from food animals in Denmark. Thus, the baseline for comparison with future prospective studies has been established, enabling the determination of trends over time.
Expression of group II phospholipase A2 in Paneth cells of an adenoma of the rectum
Authors: NEVALAINEN TIMO J. , HAAPANEN TIMO J. , RAJALA PERTTI , EKFORS TAUNO
Published: August 2009

A case of tubulovillous adenoma in the rectum of a 51‐year‐old man is presented. The tumour contained numerous Paneth cells which formed well‐developed glands in the basal areas. Group II phospholipase A2 and lysozyme were found in the tumour cells by immunohistochemistry. mRNA of group II phospholipase A2 was localized in the tumour cells by in situ hybridization. It was concluded that a considerable part of this rare type of tumour consisted of Paneth cells which were capable of synthesizing group II phospholipase A2.
Polychlorinated biphenyl (PCB) exposure produces placental vascular and trophoblastic lesions in the mink (Mustela vison): a light and electron microscopic study
Authors: BÄCKLIN BRITT‐MARIE , PERSSON ELISABETH , JONES CAROLYN J. P. , DANTZER VIBEKE
Published: August 2009

Polychlorinated biphenyls (PCBs) cross the placenta and cause fetal death in mink. No indications of impaired implantation have been reported. To study the effects of PCB on mink placental morphology, 2 groups each of 10 animals were orally exposed to Clophen A50 at 0.65 mg (low dose) and 1.3 mg (high dose) per day for 54 days, starting before mating, with 10 control animals. Placentae from mid to late gestation were examined by light and electron microscopy. In the controls, 11% of placentae were degenerate compared to 31% (low dose) and 64% (high dose) in PCB‐exposed mink. All control animals exhibited implantation sites, while one animal in the low dose and four in the high‐dose group exhibited none. However, there was no difference between PCB‐exposed and control animals in the number of placentation sites in implanted animals. Fetal death was markedly increased in PCB‐exposed mink, with only four animals (low dose) having all viable fetuses and eight (low and high dose) having a mixture of viable and dead fetuses. Nine exposed animals displayed maternal vascular lesions in the placental labyrinthine zones of viable fetuses, comprising loss and degeneration of endothelial cells, thrombi and haemorrhages. Extracellular fluid was present between the interstitial layer of maternal vessels and the syncytiotrophoblast, and there was focal degeneration of the trophoblast and fetal vasculature. It appears, therefore, that exposure of the mink placenta to PCBs affects maternal vasculature and produces degenerative changes in the trophoblast and fetal vessels, leading to fetal growth retardation or death.
An epitope shared by enterobacterial and neisserial porin proteins
Authors: HENRIKSEN ARNE Z. , MÆLAND JOHAN A. , WETZLER LEE M.
Published: August 2009

A murine monoclonal antibody (MAb F9–16) raised against a porin protein epitope called Po I of an E. coli O55 strain showed broad cross‐reactivity with bacteria within the Enterobacteriaceae, and also recognized neisseriae and moraxellae. In an immunodot assay, the antibody was bound by 32/33 strains of neisseriae and moraxellae after SDS treatment of the bacteria. Testing intact bacteria, 11/33 isolates showed definite MAb binding, including serogroup A and B meningococci. In Western blotting, the anti‐Po I MAb targeted the gonococcal porin proteins PIA and PIB, and class 1, class 2, and class 3 porins of meningococci. The MAb showed no reactivity against decapeptides which corresponded to the whole length of a meningococcal class 1 porin protein of the subtype P1, 7, 16. These findings accord with the inference that enterobacterial, neisserial and moraxellae porin proteins share an epitope (Po I) which is determined by the three‐dimensional rather than by the primary structure of the proteins and that this epitope is shielded in most isolates but surface‐exposed in some isolates, including some strains of meningococci. Since Po I is broadly distributed among commensal and pathogenic bacteria and has demonstrated immunogenicity in humans, this epitope may play a role in elicitation of “normal” antibodies with immunoprotective activity.
The 1994 human parvovirus B19 epidemic in Denmark: Diagnostic and epidemiological experience
Authors: JENSEN I. P. , SCHOU O. , VESTERGAARD B. F.
Published: August 2009

In 1994 the first human parvovirus B19 (B19) epidemic to be documented in Denmark was recorded from February 2 to September 30. In total, 10,333 serum samples were tested for specific B19 IgM and IgG antibodies, using IDEIATM Parvovirus B19 IgM and IgG kits. The prevalence of B19 IgM positivity was 11% for the whole period and 29% at the peak of the epidemic in week 14, declining from week 39 and onwards to 1–3%. The prevalence of B19 IgG (IgM‐negative samples) was 60%, indicating an earlier infection, and the same for men and women. The gender distribution of tested patients was the same at the beginning of the epidemic as at the end of the epidemic and a year after its peak, i.e. 86% of samples were from women and only 14% from men. Age distribution for women was the same for the three periods (median age 34 years). For men the median age was 32 years, 39 years and 31 years, respectively. Only a few samples from children were tested. No change in test pattern was observed during the three periods. Approximately 75% of all samples tested were from women of childbearing age (18–45 years old), suggesting a fear of fetal complications in an actual or future pregnancy, rather than a serological verification of clinical symptoms. From the sparse clinical information that accompanied the serum sample we were not able to demonstrate that women were more likely than men to have a symptomatic B19 infection. With reservations we estimate that 14% of adverse pregnancy outcome is correlated with a B19 infection.
Antibodies to pyruvate dehydrogenase in primary biliary cirrhosis: correlation with histology
Authors: KISAND KAI E. , KISAND KALLE V. , KARVONEN ANNA‐LIISA , VUORISTO MATTI , MATTILA JORMA , MÄKINEN JUDIT , UIBO RAIVO
Published: August 2009

Antimitochondrial antibodies to pyruvate dehydrogenase are the hallmark of primary biliary cirrhosis. Their pathogenic role has not been proven, although antibodies to pyruvate dehydrogenase are bound to biliary epithelium. The aim of this study was to characterize serum IgA antibodies to pyruvate dehydrogenase and to evaluate their response to different treatment regimens. We also compared the level of antibodies with severity of histological lesions and the data of biochemical liver tests. Serum samples were collected at baseline and after 24 months from 61 primary biliary cirrhosis patients, whereas 23 patients were treated with ursodeoxycholic acid, 20 with colchicine, and 18 with placebo. ELISA was used to detect antibodies to pyruvate dehydrogenase. IgA and IgG were separated with jacalin and protein‐A, respectively. Capacity of immunoglobulins to inhibit enzymatic activity was detected by spectrophotometric observation of the rate of enzyme reaction. 49 (80.3%) of the 61 patients possessed IgA antibodies to pyruvate dehydrogenase. Significant decrease in IgA antibodies was observed only in the ursodeoxycholic acid group (p7<0.05). 15 of 18 IgA preparations and all 24 IgG preparations of patients' sera were inhibitory towards pyruvate dehydrogenase (mean inhibitory percent ±SD: 42±33.4% and 79±22.4%, respectively, at a protein concentration of 100 μg/ml). The level of serum antibodies to pyruvate dehydrogenase correlated with several histological parameters (fibrosis, inflammatory infiltrate), but not with biochemical parameters. Our data reveal that IgA antibodies to pyruvate dehydrogenase inhibit enzyme function. The correlation between antibodies to pyruvate dehydrogenase and histological parameters might suggest the pathogenic role of these antibodies.
Virulence factors of Staphylococcus aureus strains causing infective endocarditis – a comparison with strains from skin infections
Authors: HOGEVIK H. , SÖDERQUIST B. , TUNG H.‐S. , OLAISON L. , WESTBERG A. , RYDÉN C. , TARKOWSKI A. , ANDERSSON R.
Published: August 2009

The objective was to study potential bacterial virulence factors in S. aureus endocarditis. S. aureus strains isolated from patients with well‐classified episodes of infective endocarditis (IE) (n=26) were compared with control S. aureus strains from consecutive patients with skin infections (n=30). The potential virulence factors studied were Staphylococcal enterotoxin A‐D (SEA, SEB, SEC, SED) and toxic shock syndrome toxin‐1 (TSST‐1) production and binding capacity to the extracellular matrix proteins: fibronectin, collagen type I, collagen type II and bone sialoprotein (BSP). None of the potential virulence factors studied was more prevalent among the IE strains. BSP binding was more often found in the control group with skin infections. Endocarditis patients with previous damage of the heart valves were more often infected by strains not producing any enterotoxin. No correlation was found between the potential bacterial virulence factors studied and IE. Concerning the toxins known to act as superantigens (SEA‐E and TSST‐1), the tendencies in this and other studies indicate that a larger study group might identify them as pathogenic factors in a subgroup of staphylococcal endocarditis.
Correlation of in vitro susceptibility testing results for amoxicillin‐clavulanate and ampicillin‐sulbactam using a panel of β‐lactamase‐producing Enterobacteriaceae
Authors: SIU L. K. , CHENG W. L. , HO P. L. , NG W. S. , CHAU P. Y. , LO J. Y. C.
Published: August 2009

Correlation between in vitro susceptibility results for amoxicillin‐clavulanate (AMC) and ampicillin‐sulbactam (SAM) was studied using 136 clinical and control strains of Enterobacteriaceae harboring TEM‐1, SHV‐1 or OXA‐1‐like β‐lactamases. Determination of minimal inhibitory concentration of antibiotics was performed by agar dilution. The β‐lactamases were initially characterized using isoelectric focusing. Further identification was done by DNA hybridization with or without prior PCR amplification. All strains sensitive to SAM were found to be sensitive also to AMC. In contrast, among those susceptible to AMC, only 50% were sensitive to SAM while 36% gave intermediate results and 14% were resistant. Major differences were found solely among SHV‐producers while minor differences occurred mostly among TEM‐producers. This phenomenon is probably related to the differential activities of clavulanate and sulbactam against various β‐lactamases. In conclusion, testing of Enterobacteriaceae isolates for susceptibility to AMC and SAM should be performed and reported individually to avoid erroneous designation of susceptibility.
Numerical aberrations of chromosome 17 in interphase cell nuclei of breast carcinoma cells: lack of correlation with abnormal expression of p53, neu and nm23 protein
Authors: SAUER TORILL , BERAKI KAHSAI , JEBSEN PETER W. , ORMEROD ELI , NÆSS ODDVAR
Published: August 2009

The genes for p53, neu (c‐erbB‐2) and nm23 are all located on chromosome 17. Abnormal expression of their protein products is an important prognostic parameter. The aim of this study was to investigate if numerical aberrations of chromosome 17 are reflected in the expression of these markers. The immunohistochemical expression was analysed on histological specimens from 33 breast carcinomas. In situ hybridization (ISH) was performed on interphase cell nuclei in air‐dried fine‐needle aspirates from the same cases using a digoxigenin‐labelled α‐satellite probe for chromosome 17. ISH for chromosome 6, 7 and 12 was used additionally to give an estimate of ploidy. Of the carcinomas 76% were aneuploid, and numerical abnormalities of chromosome 17 were found in 34%. Abnormal p53 protein was expressed in 15% (five cases). All of these were aneuploid, but only one of them revealed aneusomy of chromosome 17. Neu overexpression was found in 18% of the tumours (six cases). Five of these were aneuploid, whereas two were aneusome for chromosome 17. Four cancers showed full (normal) expression of nm23 protein, whereas 29 had reduced expression. Reduced expression was found in 23 of 25 aneuploid tumours. Numerical aberrations of chromosome 17 were found equally in carcinomas with reduced and full nm23 protein expression. Abnormal numbers of chromosome 17 seem only to have a minor impact on these markers and are not reflected significantly in their expression.
Chlamydia pneumoniae and possible relationship to asthma
Authors: LARSEN F. O. , NORN S. , MORDHORST C. H. , SKOV P. STAHL , MILMAN N. , CLEMENTSEN P.
Published: August 2009

Chlamydia pneumoniae (C.pn.) is claimed to be of importance for the development of bronchial asthma in previously healthy individuals. This is a new and speculative theory. Earlier studies have mainly focused on C.pn. and exacerbation of asthma. If this new theory were true, one would expect titres of C.pn.‐specific IgG to be higher or more common in patients compared with controls. It would also seem probable that pathobiological mechanisms as found in connection with other microorganisms could be demonstrated, i.e. presence of C.pn.‐specific IgE and the capability of C.pn. to induce or enhance histamine release from basophil leukocytes. We therefore examined C.pn.‐specific IgE, IgG and IgM in sera from 22 adults with bronchial asthma and 25 healthy controls. IgE was verified by passive sensitization of basophils from umbilical cord blood. The prevalence of IgE was approx. 69% and IgG approx. 23% in both groups. IgG‐titres were between 1:16 and 1:64 in both groups. No IgM was found. Further, C.pn. could neither induce nor enhance histamine release from basophil leukocytes of patients or controls. We conclude that patients with bronchial asthma and healthy controls do not differ in relation to 1) C.pn.‐specific IgE in sera, 2) the capability of C.pn. to induce or enhance histamine release from basophil leukocytes, since no such effect was found, or 3) previous C.pn. infection judged by the presence of specific IgG antibodies. Our results cannot support the theory that C.pn. is a cause of adult‐onset asthma.
Expression of porcine major histocompatibility antigens in cardiac tissue
Authors: MIRZAIE MASOUD , MEYER THOMAS , BERGER DANIELA , SAALMÜLLER ARMIN , DALICHAU HARALD
Published: August 2009

In this study we have used monoclonal antibodies directed against antigens of the major histocompatibility complex (MHC) class I and class II to reveal the detailed cellular distribution of swine lymphocyte alloantigens (SLA) in cardiac tissue. By applying a sensitive immunophosphatase staining reaction we detected the ubiquitous expression of SLA class I and class II on the vascular endothelium. Endothelial cells of capillaries and blood vessels were intensely stained structures in the examined swine hearts. Similar staining patterns were observed in control experiments with anti‐von Willebrand factor serum and Dolichus biflorus lectin used as immunohistochemical markers for endothelial cells. The luminal layer of studied pulmonary valves exhibited a strong staining reaction with anti‐SLA class I and class II antibodies. In contrast, normal cardiac myocytes failed to express immunodetectable amounts of either of the SLA determinants. Intercalated discs in porcine heart tissue did not react with either anti‐SLA class I or class II antibodies. Our data showing the abundant expression of SLA molecules on endothelial cells in normal swine heart may have physiological implications in cell‐mediated rejection occurring in xenotransplanted hearts.
Chromosome length polymorphism in clinical isolates of Candida parapsilosis
Authors: FERNANDO P. H. P. , SAMARANAYAKE L. P.
Published: August 2009

Chromosome length polymorphism among 24 clinical isolates of Candida parapsilosis obtained from several human sources was analysed using pulsed‐field gel electrophoresis. The isolates, from both superficial and deep infections, comprised a miscellaneous collection from the oral cavity, blood cultures, ear infections, wound secrete, a venous catheter and peritoneal dialysis fluid. Contour‐clamped homogenous field electrophoresis using a hexagonal electrode was used for pulsed field gel electrophoresis. The chromosome numbers varied from seven to nine and their sizes ranged from 0.75 to 2.6 Mb. According to the electrophoretic karyotype patterns the 24 isolates could be divided into 9 profiles. However, the majority (18 isolates) fell into 3 groups comprising 7, 8 and 9 chromosomes, containing 5, 11, and 2 isolates, respectively. The remaining six isolates, all of which were from either an oral or another superficial site of isolation, could be categorized into a further six groups. These data confirm previous observations on the genomic heterogeneity of clinical isolates of C. parapsilosis, and illustrate the possible commonality in strains from related clinical habitats.
Cryptorchidism as a caudal developmental field defect
Authors: CORTES D. , THORUP J. M. , BECK B. L. , VISFELDT J.
Published: August 2009

Cryptorchidism is a feature of abnormalities in the hypothalamo‐pituitary‐testicular axis, and almost all disorders of sexual differentiation in which a testis is present. We found cryptorchidism to be associated with malformations and dysplasias of the kidneys, the ureters and the spine from T10 to S5. The description of this association is new. The association was seen in 18% of cryptorchid boys younger than 3 years of age in a department of paediatric surgery, in 34% of cryptorchid foetuses who died in the third trimester, in 65% of cryptorchid patients with imperforate anus, and in all individuals with tritonmelia, the male variant of sirenomelia. Sirenomelia/tritonmelia is an extreme degree of abnormal differentiation of the caudal developmental field, also called caudal dysplasia, the caudal regression syndrome and the caudal regression malformation sequence. Caudal developmental field defects were also the predominant abnormalities in the other groups of patients. Thus, cryptorchidism may be a feature of abnormal differentiation of the caudal developmental field. Position and histology of the undescended testes of the patients included in the association were similar to in cryptorchidism in general. In the literature the association was reported in 5–10% of boys considered to suffer from cryptorchidism only. Furthermore, our observations are concordant with recent theories about cryptorchidism. Consequently, we propose that cryptorchidism in general may be a caudal developmental field defect. Study of cryptorchid patients exhibiting malformations or dysplasias of the kidneys, the ureters or the spine from T10 to S5 is essential in order to isolate new genetic disorders and to spot environmental factors causing cryptorchidism.
Papillary squamous cell carcinoma of the uterine cervix: diagnostic pitfalls
Authors: NAKAMURA ETSUKO , SHIMIZU MICHIO , FUJIWARA KEIICHI , YAMAUCHI HIDEAKI , MONOBE YASUMASA , HIROKAWA MITSUYOSHI , KOHNO ICHIRO , MANABE TOSHIAKI
Published: August 2009

A case of papillary squamous cell carcinoma (PSCC) of the uterine cervix is reported. The patient was a 73‐year‐old Japanese woman with acute renal failure and bilateral hydronephrosis. A cauliflower‐like mass was found in the uterine cervix. A uterine cervical biopsy specimen revealed PSCC in situ, while clinically it was an invasive carcinoma. Uterine cervical biopsy was performed a second time to confirm its stromal invasion. However, only small fragments were obtained because of heavy bleeding from the tumor and they showed PSCC in situ again. Following this, computed tomography of the pelvis revealed a 5 cm mass in the uterine cervix, invading the vagina and urinary bladder. Though deep‐wedge biopsy, loop electrosurgical excision, or cone biopsy is recommended to evaluate PSCC, it may be impossible to perform any of these procedures because of bleeding such as that seen in our case. In these circumstances, good communication between pathologists and clinicians is important since lack of communication may cause PSCC to be microscopically misinterpreted as in situ carcinoma rather than invasive carcinoma.
AgNORs in benign, borderline and transitional cell neoplasms of the urinary bladder
Authors: MISRA VATSALA , MUKHERJEE JAYATI , GUPTA SURESH C. , TANDON SATYAPRIYA , GUPTA ANIL K.
Published: August 2009

Argyrophilic nucleolar organizer regions (AgNORs) were studied in 106 tissue samples from the urinary bladder (6 normal transitional epithelium, 5 cystitis, 12 hyperplastic, 14 dysplastic lesions, 12 carcinoma in situ, 4 transitional cell carcinoma grade 0, 12 grade 1, 15 grade II and 12 grade III) to evaluate their role in differentiating benign, borderline and malignant lesions. The NOR counts presented a rising scale from normal (2.21), inflammatory (3.93 for both cystitis and hyperplasia), dysplastic (4.16), carcinoma in situ (5.08) to malignant lesions (5.28 grade I, 6.59 grade II and 8.37 grade III). It was concluded that AgNORs do not have a diagnostic role in these lesions, but that they can act as a reliable adjunct to existing parameters in the early detection of tumour recurrence and grading of malignant neoplasms.
Sarcomatoid renal cell carcinoma with chromophobe cell foci
Authors: HIROKAWA MITSUYOSHI , SHIMIZU MICHIO , SAKURAI TAKAKI , TERAYAMA KIYOMI , MANABE TOSHIAKI
Published: August 2009

We report a case of sarcomatoid renal cell carcinoma with chromophobe cell foci. The patient was a 57‐year‐old Japanese woman with a renal mass measuring 12×8×8 cm in diameter. Histologically, the tumor was composed predominantly of sarcomatoid renal cell carcinoma with foci of chromophobe cell carcinoma. The patient died of multiple metastases 2 months after operation. To the best of our knowledge, there have been only two reports of the coexistence of sarcomatoid renal cell carcinoma and chromophobe renal cell carcinoma in the literature. Our case indicates that chromophobe cell carcinoma may be connected with sarcomatoid renal cell carcinoma. In such a case, the prognosis may be related to the latter component.