bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of a Salmonella‐specific DNA probe by colony hybridization using non‐isotopic and isotopic labeling
Authors: AABO S. , THOMAS A. , HALL M. L. M. , SMITH H. R. , OLSEN J. E.
Published: August 2009

A 2.3 kilobase (kb) Salmonella probe, JEO402‐1, and two subfragments, F1214 (1.3 kb) and F1217 (0.8 kb), have been evaluated by colony hybridization using pure cultures of Salmonella serovars and non‐salmonella bacteria. JEO402‐1, and its subfragments, F1214 and F1217, hybridized to all of 156 different Salmonella serovars tested, while there was no reaction to 112 non‐salmonella strains belonging to 19 genera and 37 species of Enterobacteriaceae. Together with previously published results, the JEO402‐1 probe has now been shown to detect a total of 396 Salmonella strains belonging to 214 serovars of Slamonella subspecies I‐VI. A total of 178 non‐salmonella strains representing 23 genera and 51 species of Enterobacteriaceae have all tested negative with JEO402‐1. The hybridization results obtained using a digoxigenin‐labeled probe were similar to those obtained with 35S isotopic labeling when complete colony lysis was ensured.
Tetracycline resistance genes in Kenyan hospital isolates of Salmonella typhimurium
Authors: KARIUKI SAMUEL , MIRZA NAZIR BEGUM , WASTESON YNGVILD , SENERWA DANIEL , GATHUMA JOSEPH M. , OLSVIK ØRJAN
Published: August 2009

All 97 strains of Salmonella typhimurium isolated from patients at a hospital in Nairobi, Kenya, during 1988‐90 were resistant to tetracycline. The minimum inhibitory concentration (MIC) showed a large distribution range from 1 μg/ml to 128 μg/ml. The strains were heterogeneous with respect to plasmid content, but initially all strains possessed, in addition to other plasmids, a large 60‐, 63‐or 65‐MDa plasmid. The tetracycline resistance genes were characterized using oligonucleotide probes, and 20% of the resistant strains possessed tetracycline type A (tetrA), 6%tetr B, and 4%tetrC genes. Three strains possessed both type A and B tetracycline resistance determinants, which were shown to be located on the large 65‐MDa plasmid. There was no correlation between strains isolated from stools, blood, cerebrospinal or epidural fluids, pus, or urine, with respect to the tetracycline genotypes, MIC values or plasmid content.
Utility of an internal control for the polymerase chain reaction
Authors: URSI JEAN‐PAUL , URSI DOMINIQUE , IEVEN MARGARETA , PATTYN STEFAAN R.
Published: August 2009

The polymerase chain reaction (PCR) was used to amplify a 209 base‐pair fragment of Mycoplasma pneumoniae DNA. The amplicon was transferred into a plasmid and a 680 base‐pair piece of foreign DNA was inserted between the two amplimer sites. Plasmid DNA was added to the reaction mixture as an internal control for the polymerase chain reaction. Since the original hybridization target sites were included in this construction, one pair of amplimers could be used to amplify both the target DNA and the internal control DNA. Separation of internal control from target DNA after amplification was easily obtained on agarose gel electrophoresis. For the analysis of clinical samples with the polymerase chain reaction, the addition of internal control DNA allowed monitoring of the overall effectiveness of the amplification in each tube. With this technique approximately one‐third of the tests were shown to be unsatisfactory due to technical errors or contaminating inhibitors. Adequate internal controls are necessary to avoid false‐negative results with the polymerase chain reaction.
DNA content in primary tumours and lymph node metastases in colorectal adenocarcinoma
Authors: KOHA M. , BRISMAR B. , WIKSTRÖM B.
Published: August 2009

In 18 consecutive patients operated on for colorectal carcinoma of Dukes' stage C, the DNA patterns were determined in multiple samples of the primary tumours and in all detected lymph node metastases. Single‐cell microspectrophotometry on Feulgen‐stained smears of fine‐needle aspirates was used. When the most aggressive DNA pattern was considered representative, 12 primary tumours (67%) were designated as aneuploid. The frequency of aneuploidy among the metastases was almost the same (63%). In 15 cases (83%) the DNA patterns displayed by the metastatic lymph nodes were also found in the corresponding primary tumour, while in the remaining three cases (17%) the DNA pattern in the lymph node metastases was not seen in any of the multiple samples from the primary tumour. The observed tumour DNA heterogeneity may reflect either the multicellular origin of the tumour cells or the continuous evolution and progression of a neoplasm of unicellular origin, and may partly explain the dissimilarities between the DNA patterns of the primary tumour and the lymph node metastases. Biopsy samples from a number of metastatic lymph nodes are therefore required to ensure representativeness and to permit an adequate analysis of the prognostic role of the DNA ploidy status in lymph node metastases from colorectal cancer.
Detection of Chlamydia trachomatis antigens in first‐void urine to identify asymptomatic male carriers
Authors: GENÇ MEHMET , DUTERTRE YANN , BJÖRK MARIANNE , MÅRDH PER‐ANDERS
Published: August 2009

Early morning first‐void urine collected from 279 sexually active Swedish male recruits (mean age 19.5 years) was tested by two commercial enzyme immunoassay (EIA) kits, MicroTrak® and IDEIA III®, and by MicroTrak direct fluorescence assay (DFA), to detect Chlamydia trachomatis antigens. A result was assumed to be true‐positive when any of the two non‐culture tests were positive for the same specimen. In one case where only DFA was positive, confirmatory chlamydial testing was performed by isolating the organism from a urethral swab. On these premises, the number of true‐positive men was 26 (9.3% of all men studied). The sensitivity, specificity, positive predictive value and negative predictive value for MicroTrak EIA were 85%, 98%, 85%, and 98%, respectively. IDEIA III was less sensitive than MicroTrak EIA (42% vs 85%). In conclusion, the diagnosis of asymptomatic chlamydial infections in men can be established with reasonable accuracy by the detection of Chlamydia antigens in urine samples using MicroTrak EIA.
Squamous cell metaplasia of the bladder urothelium. A retrospective study of 36 patients
Authors: KVIST EBBE , SJØLIN KNUD‐ERIK , LAURSEN HENNING , ØRNTOFT TORBEN FALCK , STURMER MANUEL ANTONIO SANCHEZ
Published: August 2009

In a retrospective study of 28 women and eight men with squamous cell metaplasia in different parts of the bladder, including the trigone, no histopathological differences were observed among the regions. All the five (female) patients with parakeratosis had a concomitant invasive bladder tumour. Thirty‐eight % of all the patients had a simultaneous neoplastic tumour. The metaplastic lesions were investigated for keratin in 13 patients, and all were positive. In seven out of eight patients, the urothelium adjacent to the squamous cell metaplasia was also positive for keratin, indicating a direct transformation of the urothelium to squamous cell epithelium. The metaplastic cells were investigated for oestrogen receptors in five men and five women, and all were negative, suggesting no relationship between estrogens and squamous cell metaplasia of the bladder. Squamous cell metaplasia in the bladder is not considered a premalignant condition. However, metaplasia and neoplastic tumours are often associated with chronic tissue damage, and the presence of metaplasia may give a warning of conditions that can also cause cancer.
Detection and identification of Mobiluncus species by direct filter hybridization with an oligonucleotide probe complementary to rRNA
Authors: PÅHLSON C. , MATTSSON J. G. , LARSSON P‐G. , GERSDORF H. , GÖBEL U. B. , FORSUM U. , JOHANSSON K‐E.
Published: August 2009

A hybridization assay for direct detection and identification of Mobiluncus species has been developed and tested. A [32P]‐labelled synthetic oligonucleotide probe, complementary to a nucleotide sequence in the variable region V8 of Mobiluncus 16S ribosomal RNA, was utilized. One of the advantages of using rRNA as target molecule for the hybridization assays is the copy number of rRNA, which can be as high as 104, and that additionally three to six sites on the minus strand of the DNA gene can be utilized. This probe was found to be sensitive and to react with 62 of 68 tested typical or atypical Mobiluncus isolates. It was also specific, and was shown not to react with 96 tested unrelated bacterial species and isolates, including taxonomically closely related species like Actinomyces or Bifidobacterium spp., or with bacteria isolated from the vagina of both healthy persons with an undisturbed flora, as well as from patients suffering from the bacterial vaginosis syndrome (BV).
Susceptibility of strains of the Mycobacterium tuberculosis complex to fusidic acid
Authors: FUURSTED KURT , ASKGAARD DORTHE , FABER VIGGO
Published: August 2009

The activity of fusidic acid was studied in 40 strains of M. tuberculosis (of which 20 strains were mono‐ or multiresistant to standard antituberculosis drugs) and 10 strains of M. bovis. Minimum inhibitory concentration (MIC) was determined by the radiometric (BACTEC) broth method. The MIC for the 50 strains varied between 8 and 32 mg/1, with a MIC90 of 16 mg/1 for M. tuberculosis and a MIC90 of 32 mg/1 for M. bovis. Minimal bactericidal concentration (MBC, defined as the lowest concentration of fusidic acid which killed 99% or more of the population) varied between 32 mg/1 and 500 mg/1, with a MBC90 of 250 mg/1 for M. tuberculosis and 500 mg/1 for M. bovis. No cross‐resistance to other antituberculosis drugs (ethambutol, isoniazid, rifampicin, streptomycin, pyrazinamide, ofloxacin, ciprofloxacin) was observed as strains resistant to one or more standard antituberculosis drugs were as susceptible to fusidin as sensitive strains of M. tuberculosis. No synergism or antagonism could be demonstrated when fusidic acid was combined with either ethambutol, isoniazid, rifampicin or streptomycin against strains of M. tuberculosis resistant to one or more standard antituberculosis drugs. Addition of pooled human serum to the medium increased both MIC and MBC by factors of 4 and 8 at serum concentrations of 10% and 50%, respectively. Singlestep mutation to high‐level resistance to fusidic acid at a frequency of < 1.7 times 10‐8 could be readily selected at four times the MIC. These fusidic acid‐resistant organisms had a generation time 2.0–2.7 x longer than their parent organisms.
In situ hybridization: A valuable tool in diagnostic pathology
Authors: DENIJN MARYLENE , SCHUURMAN HENK‐JAN , JACOBSE KAREN C. , DE WEGER ROEL A.
Published: August 2009

In situ hybridization or hybridohistochemistry has evolved in recent years in a new histologic modality. In situ hybridization (ISH) can be used for the detection of DNA (DISH) or RNA (RISH). The potential diagnostic value within a pathologic setting are well recognized. In this review paper, we summarize the use of DISH in a pathologic setting for the detection of chromosomal aberrations and localization of DNA‐viruses like cytomegalovirus and Epstein Barr virus. RISH which is still in a more experimental stage can be applied for the localization of RNA‐virus, like human immunodeficiency virus. However, the most important application of RISH will be the detection of gene‐expression at the level of mRNA. Potentially this has many applications especially in early diagnostics of neoplastic tissues. Finally, we have summarized some pitfalls which may hamper the introduction of in situ hybridization for diagnostic purposes and some future developments in ISH.
Survival, function, morphology and serological aspects of rat renal allografts. Effect of short‐term treatment with cyclosporine A, anti‐CD4 and anti‐interleukin‐2 receptor monoclonal antibodies
Authors: STEINBRüCHEL DANIEL A. , LARSEN SVEND , KRISTENSEN TOM , STARKLINT HENRIK , KOCH CLAUS , KEMP EJVIND
Published: August 2009

The aim of this study was to investigate the effect of short‐term treatment with cyclosporine A (CyA) combined with anti‐CD4 (OX‐38) and anti‐interleukin‐2 receptor (OX‐39) monoclonal antibodies (MAbs) on graft survival, graft function, morphology, and anti‐donor antibody levels in a BN‐to‐LEW rat kidney transplantation model. Spontaneous rejection occurred at 9.3 days (range 9–10 d). Administration of CyA (12.5 mg/kg/d) for 14 days prolonged graft survival to 33 days (range 23–40 d, P<0.02). Supplementing with OX‐38 and OX‐39 100 μg/kg/d, given i.p. from days 0 to 7, further prolonged graft survival to 70 days (range 38‐> 100 d, P<0.02 vs controls and CyA group). One of seven recipients had good graft function for more than 100 days. A three‐fold increase of the MAb dosage did not improve mean graft survival (53.5 d), but three of eight recipients had well functioning grafts for > 100 days. Kidney function was characterized by reduced creatinine clearance, also in the recipients with long‐term graft survival, and a defect in concentrating urine creatinine with subsequent pronounced increase in urinary output. Graft histology showed a complex pattern of interstitial alterations including mononuclear cell infiltration, fibrosis, tubular atrophy and vascular damage with intimal/endothelial cell hyperplasia and perivascular inflammation. In nine of 10 MAb‐treated recipients with graft survival > 60 days, granular deposits of immunoglobulins and C3 were found by immunofluorescence microscopy (IFM). The deposits were localized in the glomerular capillaries and mesangium. IFM in MAb‐treated control animals could not demonstrate any deposits. Flow cytometric evaluation of posttransplant serum samples against donor target cells showed increasing amounts of anti‐donor antibodies until the time of rejection, while recipients with long‐term graft function had moderately positive cross‐matches up to two months after transplantation. Hereafter antibody titres decreased and cross‐matches at the time of sacrifice were again negative. The morphological findings and the flow cytometric cross‐match results seem to indicate a postponed antibody‐mediated type of rejection. The reason why some kidney recipients showed decreasing antibody titres and stable long‐term graft function is unclear.
Increased expression of adhesion proteins (MAC‐1) and altered metabolic response in human leukocytes exposed to surfactant in vitro
Authors: ZETTERBERG GÖRAN , CURSTEDT TORE , EKLUND ANDERS , FORSLID JAN , HED JAN
Published: August 2009

In this study the modulatory effects of a well‐defined surfactant preparation on blood leukocytes were investigated. The expression of the cell surface receptor MAC‐1 was analyzed by flow cytofluorometry, and the metabolic response was measured by a chemiluminescence technique. An increase (p<0.05) in the MAC‐1 receptor expression was observed in the granulocytes but not in the monocytes. There was a decrease in the metabolic response of the leukocytes after stimulation with phorbol myristate acetate (PMA) and a delay (p<0.01 for both) in the peak activity. Formyl‐methionyl‐leucyl‐phenylalanine (fMLP) caused an increased peak (p<0.01). Thus, the surfactant preparation had a modulatory effect on blood leukocytes with regard to the expression of the cell surface receptor MAC‐1 and the metabolic response.
Peripolar cells and other granulated epithelial cells in renal biopsies
Authors: GARDINER D. S. , MORE I. A. R. , LINDOP G. B. M.
Published: August 2009

The peripolar cell is a recently described glomerular epithelial cell which is situated within Bowman's capsule at the vascular pole. It contains cytoplasmic granules which contain plasma proteins, although it may also have a secretory function. The relationship between peripolar cells, other granulated glomerular epithelial cells and tubular epithelial cells is unclear. We have studied 242 biopsies from 19 types of renal disease for peripolar cells, other granulated epithelial cells and granulated tubular epithelial cells. Peripolar cells were most numerous in mesangioproliferative glomerulonephritis, IgA nephropathy, focal segmental glomerulosclerosis, membranous glomerulonephritis and lupus nephropathy. Other granulated glomerular epithelial cells were most prominent in diffuse lupus nephropathy, focal glomerulonephritis, acute vascular transplant rejection, crescentic glomerulonephritis and mesangioproliferative glomerulonephritis. Granulation of the tubular epithelium was most prominent in minimal change nephrotic syndrome and amyloidosis. It is likely that the granules in tubular epithelial cells represent lysosomes containing plasma proteins which have been absorbed from the tubular fluid. However, granulation of glomerular cells may represent a more specific response to glomerular damage. In addition, peripolar cells are prominent in only certain diseases, suggesting a specialized function.
The prevalence of selective IgA deficiency in type 1 diabetes mellitus
Authors: LIBLAU ROLAND S. , CAILLAT‐ZUCMAN SOPHIE , FISCHER ANNE‐MARIE , BACH JEAN‐FRANCOIS , BOITARD CHRISTIAN
Published: August 2009

A significant increase in the prevalence of selective IgA deficiency has been observed in patients with autoimmune disorders such as systemic lupus erythematosus and rheumatoid arthritis. Insulin‐dependent diabetes mellitus (IDDM) is an autoimmune disease and susceptibility to both IDDM and IgA deficiency is associated with HLA DQB1 alleles encoding non‐Asp amino acids at position 57. In order to assess whether the prevalence of selective IgA deficiency is increased in IDDM, we have screened a homogeneous series of adult patients with IDDM for selective IgA deficiency. One patient (1:261) was found to have a selective IgA deficiency. The prevalence of selective IgA deficiency among adult French blood donors is 1:1400. Thus, although IDDM and selective IgA deficiency are both associated with the presence of non‐Asp amino acids at position 57 of the HLA DQβ chain, the frequency of this immunodeficiency in adult IDDM patients is not significantly increased.
Expression of the epidermal growth factor receptor gene in human brain metastases
Authors: TORP SVERRE H. , HELSETH EIRIK , RYAN LIV , STØLAN SIGNE , DALEN ARE , UNSGAARD GEIRMUND
Published: August 2009

Biopsy specimens of human brain metastases were examined for amplification and expression of the proto‐oncogene c‐erbB1 (located on chromosome 7) encoding the epidermal growth factor receptor (EGFR). Moreover, the tumour DNA was also examined for amplification of other cancer‐related genes on this chromosome: the proto‐oncogene c‐met, the gene for platelet‐derived growth factor A‐chain, and the gene for plasminogen activator inhibitor type 1. All 18 brain metastases demonstrated positive binding of biotinylated EGF on cryosections. Three out of 18 metastases had amplification of the EGFR gene; the other chromosome‐7 genes tested were not amplified. Thus, an increased EGFR gene expression seems to be a general finding in a wide range of carcinomas metastatic to the brain, whereas we found only occasional selective EGFR gene amplifications in single cases.
Neuroendocrine differentiation in male breast carcinomas
Authors: ALM PER , ALUMETS JAN , BAK‐JENSEN ELISABET , OLSSON HÅKAN
Published: August 2009

The presence of neuroendocrine differentiation, as expressed by cellular chromogranin immunoreactivity, was investigated in paraffin‐embedded tissue material from 51 consecutive cases of male breast carcinoma. From six of these cases electron microscopic studies were included. Chromogranin‐immunoreactive cells were present in solid cords and delineated tubular structures. Ultrastructurally, dense core secretory granules could be detected. The expression of neuroendocrine differentiation was 45%, which is between two and eight times higher than reported for female breast carcinomas by other investigators. The present findings suggest that male breast carcinoma is an exclusive tumour disease showing both similarities and discrepancies when compared to its female counterpart.
Some bacterial parameters influencing the neutrophil oxidative burst response to Pseudomonas aeruginosa biofilms
Authors: JENSEN ELSEBETH TVENSTRUP , KHARAZMI ARSALAN , HØIBY NIELS , COSTERTON J. WILLIAM
Published: August 2009

Persistence of bacteria in spite of a normal host immune system and relevant antibiotic treatment is a key problem in many chronic infections, such as the bronchopulmonary P. aeruginosa infection in cystic fibrosis patients. The capability of bacteria to establish themselves in microcolonies or biofilms is an important protective mechanism of the microorganisms. We examined the human PMN oxidative burst response to P. aeruginosa in biofilm and in planktonic form. The PMN chemiluminescence response to P. aeruginosa in biofilms was reduced to 30.5–47.5% (p<0.04) and the superoxide response to 85.9% (p<0.02) of the response to equivalent numbers of planktonic bacteria. Mechanical disruption of the biofilms before the assays elicited a significantly increased response in the chemiluminescence experiments and to nonopsonized biofilms in the superoxide anion experiments. We conclude that biofilm bacteria, although able to stimulate the PMN, result in a reduced, suboptimal response leading to lack of efficient eradication of the bacteria in the chronic infection.
Neonatal Proteus meningoencephalitis
Authors: GRAHNQUIST L. , LUNDBERG B. , TULLUS K.
Published: August 2009

Proteus is an uncommon pathogen in neonatal meningitis and has, to our knowledge, not previously been described from Scandinavia. Our case illustrates the typical course of the disease when onset is within the first two weeks of life. The typical patient is a previously healthy, sometimes slightly preterm infant, who develops multiple brain abscesses and has a very poor prognosis. In cases with a later onset, factors predisposing for infection are common and the outcome is less severe. Our patient was a girl born at a gestational age of 36 full weeks, who was a little less alert than normal during the first three days and then became dramatically sick with convulsions and apnoeas. She died at the age of six days with severe brain damage.
Variable natural killer function of tumour‐infiltrating lymphocytes from breast carcinomas
Authors: GUDMUNDSDÓTTIR INGIBJÖRG , ÖGMUNDSDÓTTIR HELGA M.
Published: August 2009

Samples were obtained from 36 breast cancer patients: 24 tissue samples from carcinomas, six samples from non‐cancerous breast tissue and 22 samples of peripheral blood (PB). Natural killer (NK) activity was tested against the classical target K‐562 and the breast carcinoma cell line T‐47D, using a microcytotoxicity assay measuring inhibition of uptake of 3H‐thymidine by target cells. The active effector cells against both target cell types were identified as CD‐56‐positive large granular lymphocytes. PB lymphocytes from breast cancer patients showed normal NK activity against both K‐562 (mean inhibition: 75.4%, controls: 78.1%) and T‐47D (mean inhibition: 77.6%, controls: 78.0%). Tumour‐infiltrating lymphocytes (TIL) from 24 breast carcinomas showed considerable variation in NK activity against K‐562: only two cases (8%) had no activity (≥5% inhibition), 15 cases (63%) had moderate activity (15–48% inhibition), and seven cases (29%) showed full NK activity (defined as ≤ 50% inhibition). TIL from three carcinomas were also tested against T‐47D and gave 21–46% inhibition. The NK activity of TIL declined with time in culture with one notable exception where there was full NK activity after seven days in culture. NK activity of TIL showed significant positive correlation with increasing degree of lymphocytic infiltration of the tumour, and correlated also with PB NK activity. No correlation was apparent with stage of disease. The NK activity was usually lower in the TIL than the PB sample when tested in parallel. This difference was probably caused by experimental conditions as well as different relative numbers of natural killer cells. In six cases tested, infiltrating lymphocytes from carcinoma and normal tissue from the same breast were found to have similar NK activities. We conclude that the non‐specific cytotoxic activity of TIL from breast carcinomas covers the whole range from complete lack of function to full capacity comparable with NK activity of PB lymphocytes. The level of activity was moderately correlated with the degree of lymphoid infiltration but was not related to stage of disease. Differences in functional capacity of TIL may be of importance in predicting possible benefits from immunomodulating therapy.
IgG subclass antibodies against Helicobacter pylori heat‐stabile antigens in normal persons and in dyspeptic patients
Authors: ANDERSEN LEIF PERCIVAL , GAARSLEV KNUD
Published: August 2009

Measurement of serum IgG antibodies against Helicobacter pylori seems to be useful in the diagnosis of H. pylori infections. IgG subclass antibodies against H. pylori have, however, not been investigated thoroughly. In this study IgG1, IgG2, IgG3 and IgG4 antibody levels against H. pylori were measured using an ELISA technique in 187 normal adult persons and in 174 patients with dyspeptic symptoms, of whom 99 patients were H. pylori positive. None of the IgG subclass antibody levels were better than the total IgG level for the diagnosis of H. pylori infection. The discrimination between H. pylori‐positive and H. pylori‐negative patients was better with IgG1, IgG2 and IgG4 antibody levels than with IgG3 antibody level. IgG2 was the IgG subclass antibody that mainly contributed to the age‐dependent increase in the IgG antibody level. This sustains the suspicion that cross‐reactions between lipopolysaccharide (LPS) from H. pylori and LPS from other Gram‐negative bacteria may occur.
Comparison of HPV detection in parallel biopsies and cervical scrapes by PCR
Authors: ASK EIRIK , JENKINS ANDREW , KÆ JANNE , TROPE CLAES , KRISTIANSEN BJØRN‐ERIK
Published: August 2009

PCR was used to detect HPV DNA in cervical scrapes of 23 women with cervical cancer. Compared with PCR‐assisted HPV DNA detection in parallel biopsies, the sensitivity was 81% and the specificity 100%. We conclude that cervical scrapes can be used for detection of HPV DNA in women with cervical cancer.
Prevalence of hepatitis B and C and HIV antibodies in children in a Romanian orphanage
Authors: HAUKENES G. , BRINCHMANN‐HANSEN K. , MACOVEI O.
Published: August 2009

Two hundred and fifteen children in an orphanage in Romania were examined for serum markers of present or past hepatitis B and C virus and HIV infection. In total, 183 children (85.1%) had at least one marker (HBsAg, anti‐HBs or anti‐HBc) of hepatitis B virus infection. An HBsAg carrier state was diagnosed in 38 (20.8%) of the infected children. Among the carriers 24.3% were HBeAg carriers, 51.4% had anti‐HBe and 24.3% had neither HBe antigen nor antibody. Nine children (4.2%) had antibodies to hepatitis C virus. All sera were negative in tests for HIV antibodies. False‐positive reactions represented a considerable problem with these sera. Six percent of the sera gave false‐positive reactions in indirect ELISA tests for hepatitis C and HIV. Sera giving false‐positive reactions had rather high serum IgG levels. The results of this study indicate that these children have been heavily exposed to hepatitis B virus and to a certain degree to hepatitis C virus, while there were no cases of HIV infection in this orphanage.
Experimentally induced ileal ulcers in rats
Authors: WICKBOM GUNNAR , CHA SOON‐OK , DANIELSSON DAN , JÄRNEROT GUNNAR
Published: August 2009

We recently described an experimental model in the rat to create recurring chronic ileal inflammation including ulceration. This model is dependent on an “in vivo culture” of normal intestinal contents. In the present experimental study we examined the effect of a polymeric and a hydrolyzed formula diet on the formation of ulcerating lesions using our rat model. Two groups of rats (twenty in each group) were fed either one of these formula diets eight weeks prior to the experimental procedure and this diet was continued until sacrifice eight weeks later. Twenty control rats also underwent the experimental procedure but were fed standard rat pellets for the same time periods. At sacrifice 60% of the control rats had developed ileal ulcers. None of the rats fed the formula diets developed macroscopic ileal inflammation or ulceration. The effectiveness of formula diets in inducing remission in Crohn's disease in humans may be linked to alterations in the intestinal microflora. We hypothesize that the formula diets in this experiment exerted a protective effect against ileal ulceration by altering the ileal microflora. Preliminary studies support this hypothesis but need to be expanded.
Beta‐cell expression of 65‐kDa heat‐shock protein mRNA is function‐ and age‐dependent
Authors: PEDERSEN C. R. , JOSEFSEN K. , BOCK T. , HANSEN S. V. , BUSCHARD K.
Published: August 2009

This study examined the expression of mRNA coding for the 65‐kDa heat‐shock protein (HSP) in rat islet cells of different functional states and different ages. In addition, beta cells and non‐beta cells purified by fluorescence‐activated cell sorting were studied. Total RNA from islet cells and insulin‐producing RINm5F cells was isolated and analyzed by Northern blotting using a cDNA probe coding for the human homologue to the mycobacterial 65‐kDa HSP, after which blots were quantified by densitometric scanning. Isolated beta cells were found to express 65‐kDa HSP mRNA. The expression was increased in Lewis islet cells exposed to heat shock or high glucose concentration, four‐and three‐fold, respectively (p<0.01). In isolated beta cells cultured at high glucose concentration a doubling in the content of 65‐kDa HSP mRNA was seen compared with islets cultured at low glucose concentration (p<0.05). In islets from Lewis rats fasted for 24 h, the content of 65‐kDa HSP mRNA was 42% lower than in islets isolated from normally fed Lewis rats (p<0.01). Both in BB rats and Wistar Furth rats the content of 65‐kDa HSP mRNA was found to be higher in the 30‐ and the 60‐day‐old rats compared with the neonatal animals (p<0.01). The expression of 65‐kDa HSP mRNA was increased in RINm5F cells following heat shock, while no induction was seen after stimulation with glucose, TPA or IBMX. It is concluded that the 65‐kDa heat‐shock protein belongs to the family of inducible functional antigens in beta cells, which strengthens the interest in 65‐kDa HSP as an antigen possibly involved in the initiation of autoimmune beta‐cell destruction.
Prevalence of antibodies against heat‐stable antigens from Helicobacter pylori in patients with dyspeptic symptoms and normal persons
Authors: ANDERSEN LEIF PERCIVAL , RASKOV HANS , ELSBORG LARS , HOLCK SUSANNE , JUSTESEN TAGE , HANSEN BIRGIT FISCHER , NIELSEN CARSTEN MØLLER , GAARSLEV KNUD
Published: August 2009

Heat‐stable antigens from Helicobacter pylori were investigated for the detection of serum IgG, IgA and IgM antibodies against H. pylori by an ELISA technique. Antibody titers against H. pylori were measured in 167 dyspeptic patients, of whom 96 were H. pylori positive confirmed by culture or microscopy, and in 482 controls (0–98 years). Increased IgG antibody titers were found significantly more often in dyspeptic patients with active chronic gastritis than in patients with normal morphology, as well as in H. pylori‐positive patients as compared to H. pylori‐negative patients, independent of the endoscopic findings. The heat‐stable antigens were compared with acid glycine‐extracted antigens and a high degree of concordance was found in the results obtained with the two antigen preparations. The differences in the IgA antibody titers against H. pylori between H. pylori‐positive and H. pylori‐negative dyspeptic patients were significant and may be useful to confirm a borderline IgG result. No differences were found in IgM antibody titer between H. pylori‐positive and ‐negative patients. The greatest age‐dependent increase in IgG and IgA antibody titers was found in children, and if a lower cut‐off level is used for children than for adults, as has been proposed, the proportion of people with increased antibody titers against H. pylori would be almost constant from the age of between five and 10 years until the time between 61 and 80 years. Comparison of H. pylori IgG antibodies with IgG antibodies against Campylobacter jejuni and total antibodies against cytomegalovirus (CMV) showed a greater similarity between H. pylori and C. jejuni (R = 0.51) than between H. pylori and CMV (R = 0.22). This may possibly be caused by cross‐reactions between H. pylori and C. jejuni. The H. pylori heat‐stabile antigen seems not to be very different from other crude H. pylori antigens like acid glycine‐extracted antigens, but purification and characterization of the antigens are needed to improve antibody assays.
Expression of epidermal growth factor receptor in human meningiomas and meningeal tissue
Authors: TORP SVERRE HELGE , HELSETH EIRIK , DALEN ARE , UNSGAARD GEIRMUND
Published: August 2009

The aim of this study was to examine meningeal tissue under normal, reactive, and neoplastic conditions for expression of epidermal growth factor receptor (EGFR) using an improved histochemical method, namely biotinylated epidermal growth factor. EGFR was found in all the examined meningiomas (12 benign and three anaplastic) and in neonatal rat meninges, whereas normal and injured adult human and rat meninges did not exhibit detectable EGFR. These observations indicate that EGFR is involved in the development of meningeal tissue. Further, EGFR is abnormally expressed in meningeal tumours, indicating a role of EGFR in the neoplastic process of these tumours. The regular expression of EGFR in human meningiomas suggests EGFR as a tumour marker for this tumour type.
Immunity against diphtheria and tetanus in human immunodeficiency virus‐infected Danish men born 1950‐59
Authors: KURTZHALS JØRGEN A. L. , KJELDSEN KELD , HERON IVER , SKINHØJ PETER
Published: August 2009

To evaluate the possible need for vaccination against diphtheria and tetanus of patients infected with the human immunodeficiency virus (HIV), antibodies were measured in blood samples from 78 Danish HIV‐infected men, born 1950‐59, who could be expected to have received primary vaccination before they contracted the HIV infection. No patients (95% confidence interval: 0–4) had tetanus antibodies below the protective level, whereas 24 of the 78 patients (16–33) were unprotected against diphtheria. In the background population of the same age group and sex, 5% and 10% have been found unprotected against tetanus and diphtheria, respectively. No relationship between disease stages and antibody levels could be found. Neither was there any difference between patients with normal and reduced numbers of CD4+ lymphocytes. From 25 patients two blood samples were taken at an interval of at least one year. Anti‐tetanus titres showed a decrease comparable to that found in the background population, whereas the change in anti‐diphtheria titres was more variable with rising antibody concentrations in nine patients. The fall‐off in antibodies did not increase with progression of the disease. It is concluded that HIV‐positive younger men who have followed the vaccination program against tetanus prior to the HIV infection can be expected to be protected, whereas revaccination against diphtheria must be considered.
Melanotic neuroectodermal tumour as a predominant component of an immature testicular teratoma
Authors: ANAGNOSTAKI LOLA , JACOBSEN GRETE KRAG , HORN THOMAS , SENGELØV LISA , BRÆNDSTRUP OTTO
Published: August 2009

A case of melanotic neuroectodermal tumour (MNT), or so‐called retinal anlage tumour, as a predominant component of an immature testicular teratoma is presented. The patient was a 17‐year‐old man who furthermore had a mature mediastinal teratoma. The MNT was composed mainly of two cell types: small immature neuroblast‐like cells and large columnar or cuboidal epithelial‐like cells with or without melanin granules. The tumour cells were arranged in solid formations, nests, cords, alveolar and pseudoglandular structures with cleft‐like or glomeruloid‐like spaces. Myogenic differentiation was found in minor foci. Immunohistochemistry showed both neuroepithelial and mesenchymal features with positive staining reaction for neuron‐specific enolase (NSE), S‐100 protein (S‐100), melanoma antigen (HMB45), cytokeratin and vimentin. Vimentin, desmin and actin were present in the myoid cells. To the best of our knowledge this is the first reported case of MNT originating in the testis. As this tumour component occurred in an immature teratoma, neuroectodermal differentiation of germ cell origin is considered most likely.
Vegetable oils instead of xylene in tissue processing
Authors: RASMUSSEN BIRGITTE BRUUN , HJORT KARINA NORRING , MELLERUP INGER , SETHER GRETE , CHRISTENSEN NILS
Published: August 2009

In an effort to improve working conditions in the histopathologic laboratory, we have investigated whether less toxic substances may be substituted for the toxic organic solvent xylene currently in use. Xylene is used in the clearing as well as the deparaffinization process. We have tested whether the substitution of xylene by olive oil and coconut oil leads to any difference in the quality of the histologic sections. Two tissue blocks from each of 232 specimens sent for histopathologic evaluation were subjected to parallel processing in xylene and oil. The specimens represented a broad spectrum of tissue types. All sections were stained with hematoxylin‐eosin to permit evaluation of histologic and cytologic details. Furthermore, a range of histochemical and immunohistochemical stainings were applied to a subgroup of tissue sections. The results showed qualitative differences between xylene‐processed and oil‐processed tissue in only a minority of cases. In no cases were the oil‐processed samples considered less suitable for histologic diagnosis. In the histochemical and immunohistochemical stainings, no differences were registered. Although the long‐term stability of oil‐processed tissue remains to be clarified, we conclude that less toxic vegetable oil may probably be substituted for xylene without losing valuable diagnostic information.
Human proximal tubular cells modulate allogen responses of leukocytes in vitro
Authors: BLAEHR HANNE , ANDERSEN CLAUS B. , LADEFOGED JØRGEN , LANGHOFF ERIK
Published: August 2009

The interaction between proximal tubular cells and leukocytes was examined. In co‐cultures of tubular cells and allogenic leukocytes, tubular cells expressed MHC antigens and ICAM‐1. A small number of leukocytes adhered to tubular cells and induced cytotoxic damage. Thus, 65% of the tubular cells were viable after co‐culturing with allogenic leukocytes. These cells had low alloreactive capacity, which was not due to lack of interleukin‐1 or interferon‐gamma. The presence of tubular cells modulated the immune response of leukocytes by reducing the effect of mitogen by 80%, allogens by 65% and interleukin‐2 by 40%. A soluble factor released in the co‐cultures was a likely mediator, since addition of supernatants from co‐cultures suppressed mitogen responses by 27% compared to leukocytes cultured alone. This mediator might be prostaglandin, because addition of indomethacin to co‐cultures increased the growth response of leukocytes.
Histological examination of the rabbit middle ear in secretory otitis media with and without a ventilation tube
Authors: OVESEN THERESE , PAASKE PETER B. , BLEGVAD SUSANNE , ELBRÖND OLE
Published: August 2009

To study the time‐related histopathological lesions in secretory otitis media as it develops into the chronic stage, including the influence of ventilation of the middle ear, we examined the middle ear mucosa in a rabbit model. The observation period was from two weeks until 18 months. The histological method applied included haematoxylin‐eosin and Sirius red staining for morphometric quantification of the goblet cells, the thickness of the connective tissue layer, the area fractions of blood vessels, inflammatory cells, fibroblasts and collagen fibres. In conclusion, the middle ear mucosa in secretory otitis media exhibited inflammatory changes correlating with the duration of the disease. Ventilation of the middle ear in secretory otitis media did not improve the changes.
Hepatitis C virus antibodies and hepatitis C virus RNA in Chinese blood donors determined by ELISA, recombinant immunoblot assay and polymerase chain reaction
Authors: ZHANG YONG‐YUAN , HANSSON BENGT GÖRAN , WIDELL ANDERS , NORDENFELT ERIK
Published: August 2009

Antibodies to hepatitis C virus (anti‐HCV) were determined in Chinese blood donors from the city of Wuhan by a second generation ELISA screening test and a confirmatory recombinant immunoblot assay (RIBA II). Two materials of 281 and 222 sera were sampled under similiar conditions in 1989 and 1990, respectively. The first collection of sera was sent to Sweden in lyophilized form, the second directly as fresh unfrozen sera. A high proportion (7.1%) of the lyophilized sera reacted positively in the anti‐HCV screening assay, but only seven (2.5%) were positive by the RIBA confirmatory test. In four of these sera HCV‐RNA could be detected by polymerase chain reaction (PCR) analysis. In the second material of fresh sera six reacted positively in the screening anti‐HCV ELISA, but only one was RIBA positive and four were RIBA indeterminate. None of these sera was positive for HCV‐RNA. Thus, the overall prevalence of anti‐HCV among the 503 Chinese blood donors, as identified by RIBA, was 1.6%, and of HCV‐RNA by PCR 0.8%. The confirmed antibody prevalence is higher than reported from the Western world. Caution about using data from the screening ELISA only, especially if sera have been handled in an unorthodox way, is emphasized.
Oncogene activation in rheumatoid synovium
Authors: TRABANDT ANDREAS , GAY RENATE E. , GAY STEFFEN
Published: August 2009

Rheumatoid arthritis (RA) is a chronic systemic disorder that is dominated by the debilitating sequelae associated with the progressive destruction of articular joints. The molecular and cellular basis of rheumatoid joint destruction is characterized by an abnormal expression of oncogenes modulating cellular proliferation and the induction of lysosomal and metalloproteinases. Based on the observation that the synovial hyperplasia in RA is associated with the proliferation of transformed‐appearing synovial lining cells and an overexpression of such oncogenes, the possibility that a hitherto unknown HTLV related retrovirus is involved in the etiopathogenesis of RA is discussed.
Factors affecting quantitative assessment of Pseudomonas aeruginosa adherence to buccal epithelial cells
Authors: FERGUSON MARBETH I. , SCOTT EILEEN M. , COLLIER PAUL S.
Published: August 2009

The influence of experimental protocol on estimation of adherence of P. aeruginosa to buccal epithelial cells is highlighted. Use of membrane filtration to remove non‐adherent bacteria was significantly affected by membrane pore size and rinse volume, but was not affected by inoculum size or test isolate. Even with optimum filtration conditions, over 58% of the non‐adherent bacterial population were retained on the filters. Separation of non‐adherent bacteria was greatly improved when density centrifugation was employed, with less than 30% of non‐adherent bacteria present in the buccal epithelial suspension. Adherence values measured by a radiological and fluorometric method were significantly higher than those determined by microscopic counting. Radiological assays had the lowest variance. Gradient centrifugation was used to prepare a standard suspension of buccal epithelial cells. However, cells collected on different days showed significantly different adherence measurements. Adherence of bacteria to trypsinized buccal epithelial cells and to buccal epithelial cells from cystic fibrosis patients was similar, whereas adherence to normal buccal epithelial cells was significantly lower.
Serum levels of fetal antigen 2 in hyperthyroidism and primary hyperparathyroidism
Authors: RASMUSSEN H. BOJE , TEISNER B. , GRAM J. , BRIXEN K. , YDE‐ANDERSEN E. , BOLLERSLEV J.
Published: August 2009

Serum concentration of fetal antigen 2 (FA2) in patients with hyperthyroidism (n = 18) (median: 12.9 mAU/1; range: 3.2–22.4 mAU/1) was significantly (p<0.002) higher than in age‐ and sex‐matched healthy controls (median: 4.1 mAU FA2/1; range: 2.4–10.0 mAU FA2/1). Serum FA2 was positively correlated with thyroxine (T4) (RS= 0.51; p<0.05), triiodothyronine (T3) (RS= 0.64; p<0.01), bone‐Gla protein (BGP) (RS= 0.70; p<0.01), total alkaline phosphatase (total‐AP) (RS= 0.62; p<0.01), bone isoenzyme alkaline phosphatase (bone‐AP) (RS= 0.63; p<0.01), N‐terminal procollagen type III (PIIINP) (RS= 0.65; p<0.01) and urine OH‐proline (OHP) (RS= 0.79; p<0.01). In patients with hyperparathyroidism the pretreatment levels of FA2 (n = 8) (median: 17.6 mAU/1; range: 5.2–35.0 mAU/1) were significantly (p<0.001) higher than those of age‐ and sex‐matched controls (median: 3.7 mAU FA2/1; range: 3.4–9.0 mAU/1). The pretreatment level of FA2 was positively correlated with the parathyroid hormone (PTH) (RS= 0.80; p<0.05). Following surgical treatment the serum concentrations of FA2, PTH, and BGP decreased compared to pretreatment levels and the fall in these three parameters revealed parallelism. These data indicate that serum FA2 can be used as a marker in the evaluation of metabolic bone diseases.
Haemagglutination patterns of Helicobacter pylori
Authors: LELWALA‐GURUGE JANAKI , LJUNGH ÅSA , WADSTRÖM TORKEL
Published: August 2009

Thirty‐two Helicobacter pylori strains were screened for haemagglutination (HA) activity with erythrocytes of 11 different animal species. Twenty‐three strains (72%) that agglutinated human erythrocytes exhibited a broad‐spectrum HA profile. Human, guinea pig and bovine erythrocytes high in sialoglycoconjugates were strongly agglutinated by most strains. Except for two, seven strains (22%) that did not agglutinate human erythrocytes exhibited a narrow‐spectrum HA profile, commonly not inhabitable by sialoglycoconjugates or N‐acetylneuraminlactose (NANLac). Strains were classified into three major HA classes. HA of 10 strains (31%) in class I was inhibited by different combinations of NANLac, orosomucoid or fetuin, but not by asialofetuin, suggesting the presence of sialic acid‐specific HAs probably recognizing NeuAc α‐(2–3)‐ Gal isomer. Twelve strains (38%) in class II exhibited a different receptor specificity binding to different combinations of NANLac, orosomucoid and fetuin, as well as asialofetuin. No inhibition was observed with 10 strains (31%) in class III; thus, this receptor seems different from both the other classes. Of 21 strains (66%) in classes I and II, HA of 11 strains (34%) was inhibited with NANLac, 14 strains (44%) with orosomucoid and 15 strains (47%) with fetuin. The great heterogeneity observed in HA patterns indicates that the HAs of different strains may recognize a heterogeneous class of sialoglycoconjugates on the erythrocyte membrane.