bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation and identification of Encephatitozoon hellem from an Italian AIDS patient with disseminated microsporidiosis
Authors: SCAGLIA MASSIMO , SACCHI LUCIANO , GATTI SIMONETTA , BERNUZZI ANNA MARIA , POLVER PAOLA DE PICEIS , PIACENTINI ITALO , CONCIA ERCOLE , CROPPO GIAN PIERO , DA SILVA ALEXANDRE J. , PIENIAZEK NORMAN J. , SLEMENDA SUSAN B. , WALLACE SARA , LEITCH GORDON J. , VISVESVARA GOVINDA S.
Published: August 2009

Microsporidia are primitive mitochondria‐lacking spore‐forming eukaryotic protozoa that infect a wide variety of animals and also humans. Of the five genera (Encephalitozoon, Enterocytozoon, Septula, Nosema and Pleistophora) that cause infections in humans, Enterocytozoon bieneusi. Septula intestinulis, and Encephulitozoon hellem are being increasingly identified in patients with acquired immunodeficiency syndrome (AIDS). E. bieneusi causes gastrointestinal disease, S. intestinulis causes gastrointestinal and disseminated disease, and E. hellem causes ocular as well as disseminated disease. We have established in continuous culture a strain of microsporidia isolated from the urine and throat washings of an Italian AIDS patient and identified it as Encephalitozoon hellem, based on its ultrastructural morphology, antigenic pattern, and polymerase chain reaction‐amplified small subunit ribosomal RNA. We believe that this is the first time that a strain of microsporidia has been isolated from the throat washings of a patient with microsporidiosis.
Peritoneal interleukin‐6, interleukin‐8 and granulocyte elastase activity after elective abdominal surgery
Authors: TSUKADA KATSUHIKO , TAKENOSHITA SEI‐ICHI , NAGAMACHI YUKIO
Published: August 2009

In this study we investigated the interleukin‐8 concentration (IL‐8) and granulocyte elastase activity (GE) after elective abdominal surgery. Postoperative interleukin‐6 (IL‐6), IL‐8 concentrations and GE in the peritoneal fluid were examined in 27 patients who underwent various types of elective abdominal surgery. We compared these results with clinical parameters of surgical stress, operating time (OT) and blood loss during the operation (BL). P‐IL‐6 and P‐IL‐8 were significantly correlated with OT (P‐IL‐6; r=0.67, P<0.001: P‐IL‐8; r=0.59, P<0.001) and BL (P‐IL‐6; r=0.61, P<0.001: P‐IL‐8: r= 0.48, P<0.01). P‐IL‐8 was significantly correlated with P‐IL‐6 (r=0.68, P<0.001) and there was a positive correlation between GE and P‐IL‐8 (r=0.37, P<0.05). These findings indicate that IL‐8 might activate granulocytes in the peritoneal cavity after elective abdominal surgery and that assaying P‐IL‐6 and P‐IL‐8 is useful in assessing the host's response to surgical stress.
Immunoglobulin‐containing cells in the colonic mucosa in patients with human intestinal spirochaetosis
Authors: LINDBOE C. F. , ENGESVOLL I. , DARELL M. , KOPSTAD G.
Published: August 2009

The relative proportions of cells containing the various classes of immunoglobulins in the colonic mucosa were determined in eight patients with human intestinal spirochaetosis (HIS) and in eight controls. All specimens were taken from colonic resections performed because of adenocarcinoma. None of the cases with HIS showed an inflammatory reaction in the mucosa as judged subjectively by light microscopy. Cases with HIS had smaller proportions of IgD‐ and IgE‐positive cells and a larger proportion of IgA cells as compared with the controls, whereas the proportions of IgG and IgM cells were similar in the two groups. Taking into account the large individual variations in the proportions of immunoglobulins in both groups, our findings must be interpreted with caution. However, our results do not support the previous demonstration of increased percentages of IgE‐positive cells in patients with HIS.
Comparison of genotypic and phenotypic methods for species identification of coagulase‐negative staphylococcal isolates from blood cultures
Authors: HEDIN GÖRAN
Published: August 2009

Coagulase‐negative staphylococci are often isolated from blood cultures. Simple methods are needed for correct identification of those species most frequently found. In this study, PCR methods were developed for the identification of S. epidermidis and S. haemolyticus, based on DNA sequences in their 16S rDNA. The results obtained by these methods were compared with those obtained using a number of phenotypic methods, including two commercial kits API Staph® and Staphzyme®. Fifteen type collection strains and 133 blood culture isolates were tested. The sensitivity of the PCR for identification of S. epidermidis was 99% compared with API Staph®, but the specificity was lower, 94%, because of positive results also for S. capitis. The results by the PCR for identification of S. haemolyticus correlated closely with the Staphzyme® results, 13 isolates being identified by Staphzyme® and 16 by the PCR. API Staph®, however, identified only four clinical isolates as S. haemolyticus, probably too few. Among the individual phenotypic tests performed, a trehalose‐mannitol agar method and a desferrioxamine disc diffusion test for the identification of S. epidermidis were found to be very accurate. Anaerobic growth after overnight incubation could be used to distinguish S. epidermidis from S. hominis. The conclusion is that a majority of all Gram‐positive, catalase‐positive and coagulase‐negative blood culture isolates can be typed as regards species level using only a few genotypic and/or phenotypic tests.
DNA content and expression of PCNA and p53 in Hodgkin's disease and Hodgkin's‐like B‐ceil lymphoma
Authors: CZADER MAGDALENA , PORWIT ANNA , ÖST ÅKE , AUER GERT
Published: August 2009

DNA ploidy (by image cytometry) and expression of proliferating cell nuclear antigen (PCNA) and p53 tumor suppressor gene product (by immunohistochemistry) were investigated in 15 cases of Hodgkin's disease (HD) and 12 cases of HD‐like B‐cell lymphoma (HD‐like NHL). Reed‐Sternberg (RS) cells and their variants were DNA aneuploid in all cases. However, the fraction of hyperoctaploid tumor cells was higher in HD than in HD‐like NHL. PCNA expression was high in neoplastic cells (>50%) and variable (5–40%) in reactive lymphocytes in both HD and HD‐like NHL. p53 positivity was found in RS cells and their variants in 64% of HD cases, but only in 25% of cases of HD‐like NHL. Our results support the suggestion that HD‐like B‐cell lymphomas should be considered as highly malignant non‐Hodgkin's lymphomas rather than Hodgkin's disease.
Detection of respiratory syncytial virus in children in the 1993‐94 winter season in Izmir, Turkey, by two diagnostic methods
Authors: DERELI DENIZ , ERTEM EKIN , SERTER DEMIR , ŞADIMENT MUSTAFA , ÇOKER MAHMUT , TANAÇ REMZIYE
Published: August 2009

Respiratory syncytial virus (RSV) infection was searched for in 65 children between 2 months and 2 years of age hospitalized with the presumptive diagnosis of acute bronchiolitis, and in 35 children aged 6 months to 8 years with upper respiratory tract symptoms, by direct fluorescent antibody (DFA) test and cell culture. RSV infection was detected in 29.2% of the first and in 11.4% of the second group. The overall positivity rate was 23%. The sensitivity and the specificity of the commercially available DFA test in comparison with cell culture were 81.8% and 92.5%, respectively.
Current status of telepathology
Authors: EIDE TOR J. , NORDRUM IVAR
Published: August 2009

Telepathology is moving from the experimental stage to become a regular feature of pathology practice. This has been made possible by technical advances in telecommunications and image processing. Since 1990 the University Hospital of Tromsø has provided local hospitals in northern Norway with a remote frozen section service and with access to video conferences for the review of microscopic findings and for the discussion of major diagnostic issues. Several other hospitals in Norway are now participating in this development and practical relations among pathology laboratories for the purpose of consultation and education will be the next step in the procedure. Similar developments in telepathology have taken place in other countries. Standardization of network and telepathology workstations will be needed before extensive international collaboration can be achieved. Progress in high quality video devices, high capacity telecommunication lines and improved image compression techniques will increase the usage of telepathology services and make them cost‐effective. Thus, telepathology will contribute to the development of pathology services in the next century.
Ultrastructural localization of monoclonal antibodies against Pneumocystis carinii
Authors: SUKURA ANTTI , UKKOLA TARJA , LINDER EWERT , LINDBERG LARS‐AXEL
Published: August 2009

The ultrastructural immunolocalization of target antigens recognized by two monoclonal antibodies against Pneumocystis carinii was investigated using both human‐ and rat‐derived organisms. Labelling was seen using both cryo electron microscopy and postembedding in Epon after fixation in glutaralde‐hyde. The antibodies showed different host species‐ and developmental stage specificity. One of the antibodies reacted with the outer membrane of all developmental stages of human‐derived but not rat‐derived organisms, whereas the other reacted only with the cyst form. The distribution of the latter was similar to that reported for methenamine silver, a widely used cytochemical stain for the parasite, and reacted with both human‐ and rat‐derived organisms. The antibodies described appear to be useful markers in studies on the differentiation of Pneumocystis carinii.
Phase variation in streptococci of serological group B
Authors: SALASIA S. ISRINA OCTAVIA , WIBAWAN I. WAYAN TEGUH , LÄMMLER CHRISTOPH , SELLIN MATS
Published: August 2009

Encapsulation is thought to be a critical virulence factor in streptococci of serological group B. In the present study two encapsulated low‐density variants could be separated from their unencapsulated original strains by Percoll gradient centrifugation. The original strains had been isolated from human endocarditis and bovine mastitis. Type antigen preparations of the encapsulated human and bovine group B streptococcus reacted with type III‐ and type IV‐specific antiserum, respectively. No comparable reactions could be observed with their unencapsulated parent strains. In contrast to the original strains, the encapsulated variants grew with uniform turbidity in fluid medium and formed diffuse colonies in soft agar. The original strains grew as granular sediment and formed compact colonies in soft agar. In addition, the original strains appeared to have a more hydrophobic surface and showed significantly greater adherence to epithelial cells. In contrast to the nonencapsulated parent strains, the encapsulated variants were less phagocytosed by polymorphonuclear leukocytes. These findings may help our understanding of the pathogenic importance of phase variants in infections with this bacterial organism.
Immunoelectrophoretic characterization and cross‐reactivity of Rochalimaea henselae, Rochalimaea quintana and Afipia felis
Authors: ENGBÆK KRÆSTEN , KOCH CLAUS
Published: August 2009

The soluble antigens of Rochalimaea henselae, Rochalimaea quintana and Afipia felis were characterized by crossed immunoelectrophoresis using bacterial sonicates as antigens against pooled hyperimmune rabbit sera. A precipitin pattern was drawn for each bacterium and shown to be reproducible and stable even when normal or preimmune rabbit serum was incorporated in the intermediate gel. By this technique 56 antigens were identified from R. henselae, 49 from R. quintana, and 39 from A. felis. The serological cross‐reaction between R. henselae, R. quintana and A. felis, and between these 3 bacteria and 32 pathogenic bacteria was analysed by rocket‐line immunoelectrophoresis, crossed‐line immunoelectrophoresis, and tandem‐crossed electrophoresis. It was concluded that (i) 4–7 antigens distinguish R. henselae, R. quintana and A. felis from each other, (ii) both Gram‐positive and Gram‐negative bacteria cross‐react with R. henselae, R. quintana and A. felis antisera, (iii) the cross‐reacting antigens of Gram‐negative bacteria have both precipitating and non‐precipitating specificities, whereas Gram‐positive bacteria have mainly non‐precipitating specificities, (iv) the cross‐reacting antigens are common to several species, and (v) fewer cross‐reacting antigens are found in phylogenetically disparate species than in more closely related species.
Antibody response in rabbits infected with Rochalimaea henselae, Rochalimaea quintana and Afipia felis
Authors: ENGBÆK KRÆSTEN , KOCH CLAUS
Published: August 2009

Antibody responses in three pairs of rabbits inoculated with live Rochalimaea henselae, Rochalimaea quintana and Afipia felis were studied by enzyme immunoassay with whole‐cell and bacterial sonicates as antigen. No differences in measured antibody responses were found with the two types of antigen preparation. Two rabbits did not respond with antibody production. In the remaining rabbits there was a low‐titred antibody response that showed no significant cross‐reaction with related bacteria. After rechallenge the antibody response rose significantly and there was significant cross‐reaction with related bacteria. The antigens involved in the antibody response were examined by crossed immuno‐electrophoresis. After the initial inoculation 5–7 precipitin lines of the reference diagrams were deflected, including lines which cross‐reacted with antigens found in related bacterial species. After reinoculation several more precipitin lines were deflected, including additional lines cross‐reacting with antigens present in related bacteria and common bacterial antigens.
The true extent of ulcerative colitis?
Authors: LEIDENIUS MARJUT , KELLOKUMPU ILMO , LINDEN HANS , TASKINEN EERO
Published: August 2009

Twenty‐six radiological, endoscopic and histological examinations of the large bowel were performed in 25 patients with ulcerative colitis. Extensive colitis was observed in 42% of the radiological, in 38% of the endoscopic, and in 27% of the histological examinations. Agreement as to the extent of the colitis between all three examination methods was reached in 53% of the cases. The endoscopic extent of colitis varied between left‐sided and extensive in 41% of the patients during a 7‐year median follow‐up. The frequent change in the extent of ulcerative colitis and the considerable disagreement between radiography, colonoscopy and histology in evaluating the extent of colitis justify the question, “Does a true, unambiguous extent of colitis really exist?” It is more likely that the extent of colitis is just a function of time and the examination method used.
Toxic shock syndrome related to simultaneous Staphylococcus aureus epiglottic abscess and group A streptococcal pharyngitis with bacteremia
Authors: EJLERTSEN TOVE , PORSBORG PER ANDRÉ
Published: August 2009

A case of toxic shock syndrome (TSS) is reported in which both a TSST‐1‐producing Staphylococcus aureus strain and a group A beta‐hemolytic streptococcal strain (GABHS) were involved. The S. aureus group I strain was isolated from an epiglottic abscess, and the GABHS strain from tonsils and blood cultures. Acute intubation was needed because of obstruction of the airways by swollen and hyperemic mucous membranes in the oro‐ and hypopharynx together with external edema around the neck. Intravenous therapy with penicillin G was instituted initially on account of GABHS cellulitis and bacteremia. The patient's condition deteriorated during this treatment, and improvement did not occur until drainage and intravenous dicloxacillin therapy was instituted. It is not possible in this case to draw any conclusions as to which of the two organisms caused the TSS or if they were both involved.
Molecular and cellular aspects of the secretory immunoglobulin system*
Authors: Brandtzaeg PER
Published: August 2009

Adaptive immunological protection of mucous membranes is provided mainly by secretory IgA antibodies. Such “first line” defence is accomplished through a unique cooperation between the mucosal B‐cell system and the secretory component (SC) expressed basolaterally on glandular epithelial cells. This transmembrane glycoprotein is quantitatively the most important receptor of the immune system because it is responsible for external transport of locally produced polymeric IgA (plgA), which is the major product of humoral immunity. Transmembrane SC belongs to the Ig supergene family and functions as a general pig receptor, also mediating the external translocation of pentameric IgM to form secretory IgM. The B cells responsible for local pig production are intitally stimulated in lymphoepithelial structures, particularly the Peyer's patches in the distal small intestine, from which they migrate as memory cells to exocrine tissues all over the body. Mucous membranes are thus furnished with secretory antibodies in an integrated way, ensuring a variety of specificities at every secretory site. There is currently great interest in exploiting this integrated or “common” mucosal immune system for oral vaccination against pathogenic infectious agents. However, much remains to be learned about mechanisms for antigen uptake and processing necessary to elicit mucosal immunity as well as the molecular biology and cytokine regulation of SC‐dependent pig transport. Moreover, evidence is emerging for the existence of subcompartmentalization in the mucosal immune system, particularly a dichotomy in cellular migration between the gut and the upper airway, which may complicate the design of efficient local vaccines.
Gastric localization of Leishmania in a patient with acquired immunodeficiency syndrome
Authors: Gradoni L. , Guaraldi G. , Codeluppi M. , Scalone A. , Rivasi F.
Published: August 2009

We report a case of gastric localization of Leishmania in a 29‐year old man affected by AIDS. Gastric biopsies revealed macrophages infected with intracytoplasmic organisms attributable to Leishmania amastigotes. The authors emphasize the importance of performing random biopsies in the absence of endoscopic abnormalities.
Pathoanatomical aspects of malignant haematological disorders among Danish patients exposed to thorium dioxide
Authors: Visfeldt JAKOB , Andersson MICHAEL
Published: August 2009

The Danish Thorotrast Study was recently reestablished and improved. The cohort has been reidentified and followed up, and now comprises 1003 Thorotrast‐exposed patients. For all suspected haematological cases, cytological and histological material has been revised and malignant diseases have been reclassified. The numbers of cases of leukemia and other related haematological disorders were as follows: 16 acute myeloid leukemia (AML); 8 myelodysplasia syndrome (MDS); 1 acute lymphatic leukemia (ALL); 3 chronic myeloid leukemia (CML); 4 non‐Hodgkin's lymphoma (NHL); 2 multiple myeloma (MM); 2 myelofibrosis (MF); 2 chronic lymphatic leukemia (CLL). Except for CLL, all cases might be Thorotrast‐induced. (Expected number of leukemias: <2.5.) The findings in the German, Japanese, Portuguese and Danish studies are very similar. Some of the characteristic features include a high incidence of AML with several erythroleukemias, many cases of MDS, and a relatively low incidence of CML. In several studies of leukemia induced by alkylating agents, erythroleukemia is also described as a prominent feature. The possibility exists that a phase of relative predominance of erythroid elements in the bone marrow may be a common and not an unusual feature in the pathogenesis of these secondary leukemias. The findings are also compared with histopathological data from a Danish control group of de novo leukemia patients and from atomic bomb survivors with radiation‐induced leukemia. The relative frequency of AML is higher among the Thorotrast‐exposed patients than among the Danish control group and the A‐bomb survivors. In contrast, low relative frequencies are seen for ALL and CML in Thorotrast cases in comparison with de novo leukemia cases and A‐bomb survivors. It can be concluded that differences in relative and absolute frequency of leukemias and myelodysplastic syndrome exist not only between the irradiated populations and the unexposed control group, but even between groups exposed to low‐LET (linear energy transfer) and high‐LET radiation.
MUM‐4, a monoclonal antibody reacting with resident peritoneal mouse macrophages
Authors: Agger RALF , Rhodes JOAN M.
Published: August 2009

A new rat monoclonal antibody, MUM‐4, which recognizes a murine antigen/epitope that is absent on monocytes, strongly expressed on resident peritoneal macrophages and almost completely absent from peritoneal macrophages 4 days after an intraperitoneal injection of thioglycollate or heat‐killed Propionibacterium acnes organisms, is described. Immunocytochemistry and flow cytometry have been used to characterize the specificity of the antibody. MUM‐4 did not react with blood granulocytes, peritoneal exudate granulocytes, lymphocytes from blood or peritoneum, isolated spleen dendritic cells, or veiled cells from the thoracic duct of mesenteric lymphadenectomized mice. The MUM‐4 antibody reacted with resident peritoneal macrophages from all the mouse strains studied. MUM‐4 appears to represent a new specificity. The MUM‐4 antibody is of the rat IgG2c isotype and exhibits complement‐mediated cytotoxicity with rabbit complement. The staining achieved with MUM‐4 by FACS or immunocytochemical methods is intense on most resident peritoneal macrophages and the antibody should be a valuable addition to the panel of monoclonal antibodies available for studies on mouse macrophages.
Characterization of an erythromycin resistance (erm) plasmid in Streptococcus pyogenes
Authors: Schalen CLAES , Gebreselassie DANIEL , StÅHl STEN
Published: August 2009

Three erythromyxin‐resistant Swedish isolates of Streptococcus pyogenes, representing different T‐types, were studied. Two of the strains showed constitutive high‐level (MIC>200 μg/ml) and one showed moderate (MIC 6.4 μg/ml) resistance; the latter strain was sensitive to lincosamide and clindamycin, and resistance was not induced by erythromycin. In each of the strains, a plasmid with an estimated Mw of 17.6±0.9×106 was isolated in addition to smaller cryptic plasmids. The three plasmids pSE701, pSE702 and pSE703 had very similar restriction enzyme cleavage patterns. Novobiocin curing of the high‐level resistance strain ER559 showed the resistance to be linked to its 17.6×106 plasmid, pSE703. Furthermore, by electroporation this rather large plasmid was reintroduced into an erythromycin‐sensitive cured derivative, acquiring resistance, and the plasmid was again recovered from the transconjugant. One of the plasmids, pSE702, was shown by filter mating to be conjugative within S. pyogenes. DNA‐DNA hybridization showed that the resistance determinant of the present three isolates was related to the erm gene on plasmid pAMpl of Enterococcus faecalis but not to that of plasmid pE194 of Staphylococcus aureus. The copy numbers of pSE702 and pSE703, derived from the two high‐level resistant strains, were 11±3 and 17±5 compared to 2±1 for pSE701, derived from the moderately resistant strain, possibly accounting for the phenotypic variation observed. The plasmids pSE702 and pAMpM showed about 80% homology in DNA‐DNA hybridization tests and high similarity in their restriction maps.
Acinar cell carcinoma of the pancreas
Authors: Kuopio TEIJO , Ekfors TAUNO O. , Nikkanen VÄINÄMÖ , Nevalainen TIMO J.
Published: August 2009

Pancreatic acinar cell carcinoma is a rare neoplasm (comprising about 1% of pancreatic tumours). We studied three cases (61‐year‐old female; 42‐year‐old male; 57‐year‐old male), whose survival after diagnosis ranged from 1 year 2 months to 6 years 8 months. There were widespread metastases in each case. The tumours had acinar, trabecular and solid growth patterns. By immunohistochemistry, pancreatic acinar cell markers including carboxyl ester lipase, pancreatic secretory trypsin inhibitor and pancreatic phospholipase A2 (group I PLA2) gave a strong positive reaction in all three cases. By electron microscopy, zymogen granules were seen in the cytoplasm of the tumour cells. Immunostaining for prostate‐specific antigen was positive in all three cases. Above‐normal concentrations of pancreatic PLA2 were measured in the serum of one patient and the values decreased during chemotherapy concomitantly with the reduction in the size of the tumour mass. In conclusion, immunohisto‐chemical demonstration of the secretory products of acinar cells including the new marker pancreatic PLA2 is useful in the differential diagnosis of pancreatic acinar cell carcinoma. Determination of the concentration of pancreatic group I PLA2 in serum may be helpful in the evaluation of therapy.
The effect of H1 receptor antagonists on peripheral blood mononuclear cells, adenoid cells and primary cell lines
Authors: Holen E. , Elsayed S. , Nyfors A.
Published: August 2009

This study describes the in vitro effect of three H1 receptor antagonists (dexchlorpheniramine, terfena‐dine and loratadine) on human peripheral blood mononuclear cells (PBMC, n = 30) from allergic patients and healthy individuals. The three H1 receptor antagonists significantly inhibited antigen/mitogen‐induced PBMC proliferation in a concentration‐dependent manner. Allergen‐specific T‐cell responses in allergic individuals were similarly inhibited. The effect of the three drugs was also tested in cultures of mononuclear cells derived from adenoid tissue. The growth kinetics were investigated using spontaneously proliferating cell lines to examine whether the inhibition was caused by general toxicity. Three cell lines, HCT 8 (an ileocaecal adenocarcinoma) RPMI 8866 (B‐cell line) and 166 A2 (T hybridoma) were tested. Loratadine (<0.03 μM) and dexchlorpheniramine (<0.62 μM) altered the kinetics of HCT 8 and RPMI 8866, respectively. When testing RPMI 8866 and 166 A2, the growth‐inhibitory effect of terfenadine and loratadine could be neutralized by addition of cell culture filtrate from RPMI 8866 or 166 A2. These culture filtrates are rich in soluble low‐affinity IgE receptor (sCD23) and IgE‐binding factor (IgEBF), respectively. Our findings show that the antihistamines investigated display some non‐convential in vitro anti‐allergic properties possibly not related to their interaction with the Hi receptor. In addition, our results suggest: a) The H1 receptor antagonists used differ in their pattern of cell inhibition; b) The inhibitory effect is completely reversible at low drug concentrations.
Live tularemia vaccine but not proteins purified from Francisella tularensis can confer protection against lethal Listeria infection in mice
Authors: Belyi YURA F. , Petrosov VLADIMIR V. , Tartakovskii IGOR S. , Suchkov YURI G.
Published: August 2009

Immunization of Balb/c mice with Francisella tularensis vaccine strain 15/10 conferred significant protection against subsequent listerial infection. Since immunostimulatory activities could apparently be relevant to surface components of the bacterium, a technique for purification of cell wall proteins was developed. The scheme designed consisted of Triton X‐100 extraction with subsequent FPLC chromatography steps, and resulted in the isolation of homogeneous proteins with molecular masses of 54 kDa (pI=6.7) and 82 kDa (pI = 5.3), and partially purified 50, 85 and 100 kDa components. It was shown that immunization with isolated proteins failed to protect mice against lethal Listeria monocytogenes infection. Possible reasons for failure are discussed.
Cloning and nucleotide sequence comparison of the groE operon of Pseudomonas aeruginosa and Burkholderia cepacia
Authors: Jensen PER , Fomsgaard ANDERS , HØIby NIELS , Hindersson PETER
Published: August 2009

By alignment of GroEL amino acid sequences from four distantly related bacteria two highly conserved domains were identified. Two oligonucleotides complementary to the conserved domains were designed based on the preferred Pseudomonas aeruginosa codon usage. The primers were used in the PCR to amplify a 900‐base fragment of the P. aeruginosa groEL gene. The fragment was sequenced and the partial GroEL sequence was expanded by vectorette PCR upstream and downstream to cover the complete P. aeruginosa groE operon. The same technique was used to sequence the Burkholderia cepacia (formely Pseudomonas cepacia) groE operon and the region immediately upstream of groES. The B. cepacia groE operon is preceded by typical ‐10 and ‐35 heat shock expression signals. A total of 2041 and 2139 bp was sequenced from P. aeruginosa and B. cepacia respectively. Each revealed two open reading frames encoding two proteins with a predicted molecular mass of 10 and 57 kDa, corresponding to GroES and GroEL respectively. The GroEL proteins show an interspecies amino acid homology of 71%, and 73% with E. coli GroEL. Both GroEL proteins are 52% homologous to the corresponding human mitochondrial GroEL protein. The sequence data confirm the existence of highly conserved structures, which could be functionally important for the concerted action of GroEL and GroES in the folding and assembly of other proteins, and possibly in the initiation of autoimmune diseases.
An analysis of antibodies against Aspergillus jumigatus in bovine serum by iminunoblotting and enzyme‐linked immunosorbent assays
Authors: Jensen HENRIK ELVANG , LatgÉ JEAN‐PAUL
Published: August 2009

Serum samples from 20 cows with spontaneously acquired systemic aspergillosis and serum collected consecutively from a cow with experimentally induced aspergillosis were assayed for antibodies against Aspergillus fumigatus in immunoblotting and enzyme‐linked immunosorbent assays. Purified antigens (18 kDa protein and 20 kDa galactomannan) from A. fumigatus as well as complex preparations of metabolic and somatic antigens were used. Antibodies against A. fumigatus were found in both infected and non‐infected cows. High titers and especially an increase in titer in consecutive samples signified infection. Reactivity with the 22 and 32 kDa antigens in preparations of exocellular antigens and the 19, 20, and 39 kDa antigens from intracellular antigens was seen with sera from infected cows only, and is therefore indicative of infection.
Interferon‐γ and interleukin‐4 production by human T cells recognizing Leishmania donovani antigens separated by SDS‐PAGE
Authors: Bahrenscheer JESPER , Kemp MICHAEL , Kurtzhals JØRGEN A. L. , Gachihi GEORGE S. , Kharazmi ARSALAN , Theander THOR G.
Published: August 2009

Crude preparations of Leishmania donovani proteins were separated by preparative SDS‐polyacryl‐amide gel electrophoresis. Fractions of separated proteins were recovered by electroelution directly from the gel into separate chambers. The isolated protein fractions were tested for induction of proliferation and interferon‐γ (IFN‐γ) production in cultures of peripheral blood mononuclear cells (PBMC) from individuals who had recovered from visceral leishmaniasis caused by L. donovani. The release of interleukin‐4 (IL‐4) by PBMC stimulated with the isolated L. donovani antigen fractions was measured after treatment with phorbol‐myristate‐acetate and ionomycin. The cells proliferated in response to all protein fractions with molecular weights in the range <12 kDa to 85 kDa. In general, IFN‐γ was secreted in response to stimulation with all the protein fractions, whereas IL‐4 production was infrequently observed. The results show that T cells from individuals who have been cured of visceral leishmaniasis recognize and respond to a wide range of leishmanial antigens. There was no evidence of particular fractions constantly giving either IFN‐γ‐ or IL‐4‐producing responses.
Chemotaxis of human osteoblasts
Authors: Lind MARTIN , Deleuran BENT , Thestrup‐Pedersen KRISTIAN , SØBalle KJELD , Eriksen ERIK F. , BÜNger CODY
Published: August 2009

The in vitro chemotactic response of human osteoblasts was investigated towards the following growth factors: TGF‐β, PDGFs, FGFs and IGFs. Human osteoblasts grown from trabecular bone after enzymatic digestion were studied. TGF‐β stimulated the migration of human osteoblasts in a dose‐dependent manner with a four‐fold increase in migrated cells at 100 pg/ml, which was the optimum concentration. PDGF‐BB also stimulated migration four‐fold in a dose‐dependent manner with a maximum response at 10 ng/ml. PDGF‐AA, IGF‐I and IGF‐II stimulated migration two‐fold at 100 ng/ml. The results show that TGF‐P and PDGF‐BB are important regulators of human osteoblast migration, but other growth factors IGF‐I, IGF‐II and PDGF‐AA may also stimulate osteoblast migration. Our results additionally suggest that TGF‐P and PDGF‐BB may participate in the recruitment of osteoblasts during bone remodeling since both TGF‐P and PDGF‐BB are found in bone matrix and could be released during osteoclastic bone resorption. They furthermore support a possible use of TGF‐P and PDGF‐BB in growth factor‐induced osteogenesis.
Strain differentiation of Neisseria meningitidis by small‐fragment restriction endonuclease analysis (SF‐REA)
Authors: Mylvaganam HAIMA , Gilja ODD HELGE , Halstensen ALFRED , HØIby E. ARNE , Digranes ASBJØRN , Bjorvatn BJARNE
Published: August 2009

Following an outbreak involving 3 cases of serogroup B meningococcal disease in a rural part of Western Norway, 2 clinical and 99 carrier isolates of Neisseria meningitidis were examined by small‐fragment restriction endonuclease analysis (SF‐REA) using EcoRI, to determine its potential for strain differentiation. SF‐REA characterized all isolates and provided reproducible results with acceptable inter‐clonal differentiation. The results of SF‐REA correlated well with those of serological typing and were used to determine clonal diversity and prevalence of invasive strains among the carrier isolates. SF‐REA was also useful in demonstrating acquisition of a new carrier strain after eradication of the initial strain by ofloxacin. Thirty‐one different restriction patterns/pattern complexes were recognized among the 101 isolates. The two clinical isolates had identical restriction patterns and showed 5:90% similarity to those of six carrier isolates. In three out of six apparent treatment failures, successful eradication of the original strain by ofloxacin was demonstrated by SF‐REA. SF‐REA proved valuable in strain differentiation of Neisseria meningitidis, complemented serology, and characterized all isolates which could not be typified by serology.
Quantification of neutrophil chemotaxis: A comparison of stereological and 51Cr‐labelling methods
Authors: Storgaard M. , West M. J. , Nielsen S. L. , Obel N.
Published: August 2009

A stereological method has been used to estimate the number of migrated polymorphonuclear leukocytes in a multipore filter assay. The estimates have been compared to those obtained with conventional 51Cr‐labelling methods. The precision of the estimates in individual subjects with the two methods was similar, and in both cases the contribution to the interindividual variance made by the methods was small compared to the biological variance. However, the 51Cr‐labelling method resulted in a significant reduction in the fraction of neutrophils that migrated as well as a delay in the chemo‐tactic response. Optimal assay conditions with regard to sample size, cell concentration, incubation time and chemoattractant fMLP concentration have been established for the stereological method and the virtues of the two techniques discussed.
Neonatal treatment of BB rats with sulphatide delays development of diabetes but does not change incidence
Authors: Buschard KARSTEN , Hageman IDA , Hansen AKSEL KORNERUP , Fredman PAM
Published: August 2009

Sulphatide is a newly described autoantigen in insulin‐dependent diabetes mellitus; it is present in the islets of Langerhans, and anti‐sulphatide antibodies are found in diabetic patients and in spontaneously diabetic BB rats. The aim of the study was to treat neonatal BB rats with sulphatide in order to induce tolerance and thereby possibly influence later diabetes development. One hundred and twelve newborn BB rats, divided into three groups, were treated once daily during the first 6 days of life with intrathymic injections of sulphatide, galactosyl‐ceramide (which is similar to sulphatide but without sulphate) or phosphate buffer alone. Although the results showed no difference in diabetes incidence among the three groups, there was a delayed onset of diabetes in the sulphatide‐treated group, which developed diabetes on average at 77 ± 1 days of age, compared to 70 ± 2 days (p<0.02) for the galactosyl‐ceramide‐treated group and 70±1 days (p<0.01) for the buffer‐treated group. The degree of insulitis and the size of islets of Langerhans were studied histologically for the diabetic animals in all three groups; there were no significant differences although the sulphatide‐treated group tended to have a more normal histology. The blood glucose levels for the diabetic BB rats were similar in all three groups. Thus, neonatal treatment with the diabetic autoantigen sulphatide at the chosen dosage does not influence the incidence of diabetes, but delays the onset of disease.
Direct identification of Staphylococcus aureus in blood cultures by detection of the gene encoding the thermostable nuclease or the gene product
Authors: Brakstad ODD G. , MæLand JOHAN A.
Published: August 2009

This study compares methods for direct identification of S. aureus in blood cultures by detection of the thermonuclease (TNase) of this bacterium or the nuc gene encoding it. The protein was detected by an enzyme diffusion test in o‐toluidine blue DNA agar with a test time of at least 4 h, by a monoclonal antibody (MAb)‐based sandwich enzyme‐linked immunosorbent assay (sELISA) with a test time of ˜4 h, and by a MAb‐based sandwich enzyme‐linked immunofiltration assay (sELIFA) with a test time of 25–30 min (sample preparation included). The nuc gene was amplified by a polymerase chain reaction (PCR) with a test time (amplification plus detection) of ‐3.5 h. The tests were optimized for direct examination of blood‐containing cultures. All tests were positive with 67/67 blood cultures which grew S. aureus, negative with 35/35 cultures which grew coagulase‐negative staphylococci, and negative with 37/37 cultures with various other bacteria. These results showed positive agreement with those of the commercial AccuProbe test but not with the StaphAurex agglutination kit. With an artificially seeded blood culture, minimum total times required (incubation plus testing) were as follows: nuc‐PCR, 9.5 h; sELIFA, 12.5 h; enzymatic test, 16–36 h; AccuProbe, 14 h. Direct examination of both the nuc gene and the mecA gene encoding methicillin resistance demonstrated the mecA gene in all the coagulase‐negative staphylococci (48.6%) which showed oxacillin resistance. The sELIFA had the particular advantage of its short test time, the PCR its high sensitivity and the possibility of simultaneous detection of the species‐specific nuc gene and genes encoding other clinically important characters of the bacteria. These tests offer the prospect of direct application to a variety of clinical specimens for rapid diagnosis of S. aureus infection.
IgA and mucosal defense
Authors: Lamm MICHAEL E. , Nedrud JOHN G. , Kaetzel CHARLOTTE S. , Mazanec MARY B.
Published: August 2009

The traditional role of IgA antibodies in mucosal defense has been considered as providing an immune barrier to keep exogenous substances, including microbial pathogens, from penetrating the mucosa. In this way infections can be prevented. More recently, studies in vitro and in vivo are providing evidence to suggest that IgA may have additional roles in mucosal defense. For example, during their passage through the lining epithelial cells of mucous membranes en route to the secretions, IgA antibodies may have an opportunity to neutralize intracellular pathogens like viruses. Also, IgA antibodies in the mucosal lamina propria have opportunities to complex with antigens and excrete them through the adjacent mucosal epithelium, again by the same route to the secretions that is taken by free IgA. These latter functions could aid in recovery from infection.
Prevalence of antibodies to Borvelia burgdorferi in Danish rodents
Authors: Frandsen FLEMMING , Bresciani JOSÉ , Hansen HANNA G.
Published: August 2009

Six rodent species from six Danish localities were examined for antibodies to Borrelia burgdorferi by immunofluorescence assay (IFA). A total of 1097 specimens were tested using a serum titre of 1:8. The wild mice Apodemus sylvatieus and A. flavieollis had high prevalences of B. burgdorferi antibodies of 42.1% and 27.9% respectively, but the vole Mierotus agrestis also showed an exceptionally high prevalence of 32.7%. Clethrionomys glareolus had a low average prevalence of 17.4%. The lowest and highest prevalences of rodents seropositive for B. burgdorferi of 6.5% and 100% were found for Mieromys minutus and Mus musculus respectively. Twice as many of the Microtidae voles (M. agrestis and C. glareolus) as of the family Muridae were caught, and in Denmark the actual number of seropositive Microtidae was 1.6 times higher than for Muridae. The results indicate that in Denmark the family Microtidae species plays an important role as an animal reservoir host for B. burgdorferi. The prevalence of antibodies against B. burgdorferi varied from locality to locality, but seropositive rodents were found at all localities examined, indicating that the spirochete was widely present in Denmark.
Shigella flexneri adherence to and multiplication in coxsackie B1 virus‐infected HEp‐2 cells
Authors: Modalsli KRISTIN R. , Bukholm GEIR , Holberg‐Petersen MONA , DegrÉ MIKLOS
Published: August 2009

Coxsackie B1 virus infection enhances the susceptibility of in vitro cultured HEp‐2 cells to invasiveness by Shigella flexneri. We have studied the effect of viral infection on two phases of the invasiveness. Only a minor part was mediated by enhanced bacterial adherence to the cells, and the intracellular multiplication was unaffected by the virus. Enhanced adherence was not dependent on the presence of the gene product of the 140 Md virulence associated plasmid. Our data indicate that enhanced invasiveness induced by viral infection is mediated by an effect on other phases of the invasiveness.
Histology of chronic pilonidal sinus
Authors: SØNdenaa K. , Pollard M. L.
Published: August 2009

Many theories concerning the development of chronic pilonidal sinus have been proposed. A histologic study of primary pilonidal sinus in 53 patients is presented. Subcutaneous tissue contained sinuses surrounded by chronic inflammation. Hair in sinuses was found in three quarters of the specimens examined. Examination showed that hair entered via one of the sinus openings created. Pits (denned as darker spots of varying width in the midline of the internatal cleft) were found to be indentations of the skin containing keratin plugs and debris. They may be isolated or connected with hair follicles. Pilonidal sinuses are chronic inflammatory processes of the skin caused by keratin plugs and debris clinically observed as pits, having penetrated the dermis.
Significance of inflammatory cervical smears
Authors: Parashari ADITYA , Singh VEENA , Gupta MADAN M. , Satyanarayana L. , Chattopadhya D. , Sodhani PUSHPA , Sehgal ASHOK
Published: August 2009

Two hundred and fifty‐seven women attending a Maternal and Child Health Centre (MCH) were examined for different colposcopic and histological patterns associated with cervical inflammation as detected by cytology and for their association with different gynaecological infections. The cytodiag‐nosis revealed inflammation in 207 women (80.5%) and non‐inflammation in 49 (19.5%); one smear was inadequate for evaluation. Fifty‐six per cent of the women with inflammation and 20% with non‐inflammation had an atypical transformation zone (ATZ), the risk of ATZ being 4.9‐fold higher in those with inflammation. Biopsies from 128 women with abnormal colposcopy revealed morphological changes suggestive of human papillomavirus (HPV) in 89 (69.5%) and dysplasia of varying grades in 8 (6.3%). Seventy per cent of histologically diagnosed HPV lesions stained immunohistochemically, whereas 84% reacted with a biotinylated Pan‐HPV probe by DNA in situ hybridization (DISH). In addition to HPV, chlamydia (OR 15.6, 95% CI 2.2, 311.6), T. vaginalis (OR 18.4), bacterial vaginosis (OR 24.7, 95% CI 3.5, 492) and herpes simplex virus (OR 4.9, 95% CI 1.4, 20.9) were significantly associated with inflammatory smears. Of 11 dysplasias detected by colposcopy and confirmed by biopsy, 8 (72.7%) had inflammatory cytology in the initial Pap smears. Thus a large proportion of women with inflammatory smears had multiple gynaecological infections and may be at increased risk of developing preneoplastic or neoplastic changes. Furthermore, they risk transmitting the infections to their partners.
The prognostic significance of immunohistochemically detectable metallothionein in primary breast carcinomas
Authors: Haerslev TORBEN , Jacobsen GRETE KRAG , Zedeler KARIN
Published: August 2009

Metallothionein (MT) is a low‐molecular‐weight protein with a high affinity for group II metal ions, especially zinc and copper. MT serves as an intracellular reservoir of these ions, but may also be involved in the detoxification of certain toxic metalions such as cadmium. In addition, high MT contents might protect tumour cells from alkylating agents and irradiation. The aim of this study was to evaluate the prognostic significance of immunohistochemically detected MT overexpression in patients with primary breast carcinoma: 478 patients with primary breast carcinoma diagnosed during the period 1980–1985 were included. Formalin‐fixed and paraffin‐embedded tissue was used. Immuno‐reactivity for MT was found to be independent of the length of formalin fixation if the sections were microwave processed before incubation with the primary antibody. Patients were divided into two groups: those with MT overexpression (more than 10% positive tumour cells) and those with low expression (less than 10% positive tumour cells). MT overexpression was correlated with postmenopausal status, large tumour size, presence of lymph node metastases, high number of mitoses, severe nuclear pleomorphism, high histological grade (poor differentiation), and absence of PgR. In univariate analysis of survival data, MT overexpression was a predictor of poor overall survival in the entire group of patients. In multivariate analysis, MT overexpression failed to be of prognostic significance, whereas classical histopathological parameters such as tumour size, histological grade, and PgR were of independent prognostic significance.
An easy microtiter assay for quantitation of cytokine induction by lipopolysaccharide (LPS) and activity of LPS‐binding serum components
Authors: Garde ANNE HELENE , Fomsgaard ANDERS , HØIby NIELS
Published: August 2009

Lipopolysaccharide (LPS, endotoxin) is a major inducer of cytokines, such as interleukin 1 (IL1), IL6, IL8 and tumor necrosis factor (TNF). A convenient microtiter assay was developed to measure such activity. LPS coated onto a plastic surface was used to stimulate purified human mononuclear cells (MNC) in microtiter plates. Following stimulation the supernatants were assayed for presence of TNF by ELISA. Purified rough and smooth LPS from Pseudomonas aeruginosa gave a dose‐dependent TNF release over a range of 0.1–1.0 μg LPS/well. The assay was subsequently used to investigate the biological activity of anti‐LPS antibodies and other LPS‐specific serum components in sera from patients with cystic fibrosis (CF). As a group, sera from 10 CF patients chronically infected with P. aeruginosa did not affect the LPS‐induced TNF release, while sera from normal controls inhibited this biological activity. When individual CF patients with or without chronic lung infection are considered, the antibodies appear to either enhance or inhibit the LPS‐stimulated TNF release (range: 73–120%), while all antibodies from healthy controls inhibit the activity of LPS (range: 76–97%). Only a weak correlation (rho=0.491, p=0.037, n=19) was found between the antibody titer in ELISA and the biological activity of sera. This new assay is suggested for convenient measurement of interference with cytokine induction from human MNC by patient or therapeutic anti‐LPS antibodies and other LPS‐specific serum components.