bluuebunny/crossref_metadata_2025 huggingface dataset:
Radiation resistance of microorganisms on unsterilized infusion sets
Authors: Christensen EBBE AHRENSBURG , Kristensen HANNE , Hoborn JAN , Miller ARNE
Published: August 2009

Three different methods were used for detecting and isolating microorganisms with high radiation resistance from the microbial contamination on infusion sets prior to sterilization. By all three methods, microorganisms with a radiation resistance high enough to be a critical factor in a sterilization process (D‐6 value ≥ 30 kGy) were found with a frequency of approximately two colony forming units (cfu) per 100 product items, even though the product items in two of the series of analyses were irradiated with doses of 3–6 kGy. The frequency of occurrence of isolates with D‐6 values > 30 kGy was 0.45 per 1000 cfu of the total aerobic count. Eight different isolates of microorganisms had D‐6 values > 40 kGy when irradiated in dried laboratory preparation. All but one of these were classified according to morphologic criteria as Deinococcus, and all but one had nonlinear dose‐response relationships in semilogarithmic presentation.
Adenocarcinoma of the vermiform appendix. A population study
Authors: Nielsen GUNNLAUGUR PETUR , Isaksson HELGI J. , Finnbogason HANNES , Gunnlaugsson GUNNAR H.
Published: August 2009

We report seven cases of adenocarcinoma of the vermiform appendix occurring in Iceland during 1974–1989. The patients ranged in age from 25–83 years, mean age 55.1 years. There were five males and two females. Five had mucinous adenocarcinoma, two had adenocarcinoma. Four patients presented with symptoms and signs of acute appendicitis and all had surgically resectable disease. Three of these patients were alive with no evidence of disease four months, two years and 15 years after presentation; one death of disease occurred seven years after ileocecal resection. In three cases, the clinical presentation was that of metastatic adenocarcinoma of unknown origin. Of these patients two were diagnosed at autopsy and one after appendectomy for perforated appendicitis. Survival in this group was six weeks, three months and twelve months, respectively. In none of our patients was the diagnosis made preoperatively and no tumors were found in appendices removed incidental to other intra‐abdominal operations. The incidence of adenocarcinoma of the vermiform appendix in Iceland during 1974–1989 was approximately 0.2 cases/100.000/year.
Detection of genital papillomavirus types by polymerase chain reaction using common primers
Authors: Jenkins ANDREW , Kristiansen BJØRN‐ERIK , Ask EIRIK , Oskarsen BENTE , Kristiansen EWY , Lindqvist BJØRN , Trope CLAES , KjØRstad KJELL
Published: August 2009

We describe the detection of eight genital human papillomavirus (HPV) types, including HPV16 and HPV18, by PCR amplification of a 323 base‐pair region of the genome within the LI open reading frame (ORF). The primer sequences are: TGYAAATATCCWGATTWTWT and GTATCWACMA‐CAGTAACAAA. The method will detect purified HPV16 DNA down to a concentration of as little as a single molecule in 100 μl. The method is also applicable to purified DNA and crude lysates from tumour biopsies. Typing of the PCR product can be achieved with specific oligonucleotide probes.
Serum amyloid A levels in acute graft‐versus‐host disease in bone marrow transplant recipients
Authors: Leskinen RITVA , Maury C. P. J. , Teppo ANNA‐MAIJA , Volin LIISA , Ruutu TAPANI , HÄYry PEKKA
Published: August 2009

We studied serum amyloid A levels of 31 bone marrow transplant recipients with and without acute graft‐versus‐host disease. Before transplantation the mean SAA concentration was 5.1±0.8 mg/1 (mean ± SEM). It remained low in patients with no signs of aGVHD but increased significantly during aGVHD to 54.0 ± 8.2 mg/l (p < 0.001 compared to pre‐BMT value). Severe infections also induced high SAA levels.
Hormone replacement therapy prevents coronary artery disease in ovariectomized cholesterol‐fed rabbits
Authors: Haarbo JENS , Hansen BIRGIT FISCHER , Christiansen CLAUS
Published: August 2009

The prevention of coronary artery disease in women is of considerable importance. We have therefore investigated the influence of oestrogen monotherapy and oestrogen‐progestogen replacement therapy on coronary artery disease using a simple morphometric method. We studied sixty‐three cholesterol‐fed rabbits for nineteen weeks. They were randomized to either ovariectomy (51 rabbits) or a sham operation (12 rabbits). The ovariectomized rabbits were randomized to receive either 17 β‐estradiol, 17 β‐estradiol plus norethisterone acetate, 17 β‐estradiol plus levonorgestrel, or placebo. The rabbits with the sham operation received placebo. The hormone therapies reduced the development of coronary artery disease compared to placebo (p<0.0001). Furthermore, the coronary artery disease was attended by atherosclerosis in the more distal parts of the coronary arteries (p< 0.0001), the thoracic aorta (p<0.0001) and the abdominal aorta (p<0.0001), and by a reduced relative heart weight (p<0.05). We conclude that coronary atherosclerosis can be determined quantitatively by morphometry in rabbit arteries. Estradiol monotherapy reduces coronary atherosclerosis in cholesterol‐fed rabbits and the addition of norethisterone acetate or levonorgestrel does not attenuate this beneficial effect.
Antimicrobial resistance of enteropathogenic Escherichia coli strains from a nosocomial outbreak in Kenya
Authors: Senerwa DANIEL , Mutanda LABIUS N. , Gathuma JOSEPH M. , ØLsvik ØRJAN
Published: August 2009

The majority of the 78 enteropathogenic (EPEC) and the 151 non‐EPEC Escherichia coli strains isolated from preterm neonates during an outbreak of gastroenteritis in a hospital in Nairobi, Kenya, were resistant to trimethoprim‐sulfamethoxaxole, chloramphenicol, oxytetracycline and ampicillin, but only a few strains were resistant to cefazolin, cefamandole, cefotaxime, amikacin and nalidixic acid. Fourteen different antimicrobial resistance patterns were observed in the 229 strains of E. coli analysed. Eighty‐two percent of the EPEC strains belonged to two resistance patterns compared with 79% of non‐EPEC strains which exhibited three resistance patterns. There was no consistent relationship between plasmid profile group and antimicrobial resistance pattern, although one resistance pattern was more frequently observed in EAF‐positive strains belonging to the dominant plasmid profile group. Nine percent of the EPEC strains were resistant to gentamicin compared to 37% in the non‐EPEC group. No correlation was observed between administration of gentamicin and percentage of resistant strains isolated. None of the nine neonates receiving gentamicin died during the outbreak. Gentamicin resistance was observed in E. coli strains from six out of these nine neonates. Five out of fourteen neonates who received other antimicrobials, or no antibiotic treatment at all, died.
Prognostic implications of image cytometric assessments of nuclear DNA distribution pattern of neoplastic cells in thyroid medullary carcinoma
Authors: Soares JORGE , Fonseca ISABEL , Limbert EDWARD , Falkmer URSULA G. , Falkmer STURE
Published: August 2009

A modified technique for cytometric analysis of the nuclear DNA distribution pattern of neoplastic cells has been applied on archival histopathological specimens originating from 42 patients who had undergone thyroidectomy for medullary carcinoma of the thyroid gland. Five of the cases were of familial type. The DNA cytometric assessments were made by means of computerized image analysis techniques on Feulgen‐stained, intact, cytodiagnostically identified neoplastic nuclei obtained from histopathologically selected areas of paraffin blocks. The nuclei were enriched by means of a cytospin technique after deparaffinization of pronase‐induced disaggregation of 50 urn thick sections. Only seven of the tumours were found to consist of neoplastic cells where the nuclei showed a DNA distribution pattern of “aneuploid” type; six of these patients had a rapidly progressive neoplastic disease, but the seventh patient did not. Among all the patients whose tumour cell nuclei showed a cytometric DNA ploidy pattern of “euploid” type, not less than about half had a rapidly progressive neoplastic disease. Thus, even when a refined cytometric technique is used, the value of the nuclear DNA ploidy pattern of the neoplastic cells as a prognostic variable in MTC is limited.
Wernicke's encephalopathy in an autopsy material obtained over a one‐year period
Authors: Vege ÅSHILD , Sund STÅLE , Lindboe CHRISTIAN FR.
Published: August 2009

In a material comprising 279 consecutive autopsies obtained over a one‐year period there were four cases of Wernicke's encephalopathy (WE), of which three were inactive (chronic) and one was active (acute). The latter was not related to chronic alcoholism. Our findings show that WE may be regularly present in a routine autopsy series. In only one case was WE suspected clinically and none of the cases revealed macroscopic brain changes indicative of WE. We therefore suggest that sections of the mammillary bodies should be taken routinely to detect all possible cases of WE.
Interleukin‐2 stimulation of blood mononuclear cells from patients with inflammatory bowel disease
Authors: Tvede NIELS , Brynskov JØRN
Published: August 2009

The functional capacity of biologically active, high‐affinity interleukin‐2 receptors (IL‐2R) was studied by means of interleukin‐2 (IL‐2) stimulation of blood mononuclear cells (BMC) from 22 patients with inflammatory bowel disease (IBD) and 24 controls. The spontaneous, as well as the IL‐2‐induced, proliferative responses were significantly decreased in patients with active IBD, whereas the expressions of biologically inactive, low‐affinity IL‐2R (i.e. TAC antigen or CD25) were significantly increased in the same BMC cultures. In contrast, no significant differences were seen between patients and controls when BMC were stimulated with a nonspecific mitogen (phytohemagglutinin). The results suggest that a downregulation of IL‐2 responsiveness may contribute to decreased BMC proliferation in vitro in active IBD.
Central nervous system calcifications following tap water rinsing in autopsy material from children
Authors: Reske‐Nielsen EDITH , Lund ELISABETH , Gregersen MARKIL
Published: August 2009

CNS from 80 children collected consecutively at the Institute of Forensic Medicine, Aarhus, were investigated. Owing to peculiar calcifications of the first 41 CNS examined, we gradually came to suspect that some external factors had influenced the results. Different fixation methods used on the CNS, rinsed or not rinsed in tap water, led us to suggest that the relatively high calcium content in the tap water in our part of the country was the cause. The non‐pathologic calcification could presumably mimic metabolic diseases.
The internal elastic lamina in the basilar artery and its possible significance for intimal thickening
Authors: Heldal K. , Svendsen E. , MæEhle B. O.
Published: August 2009

The degree of intimal thickening, folding of the internal elastic lamina (IEL) and luminal radius/wall thickness ratio (RWR) have been compared between predilectional sites and a non‐predilectional site for atherosclerosis in human basilar arteries. The findings are compatible with the hypothesis that a low degree of folding and a high RWR facilitate intimal thickening. A negative correlation between the fold index and intimal thickening index and a positive correlation between RWR index and intimal thickening index were found. Predilectional areas also have lower fold indices and higher RWR in younger subjects prior to any intimal thickening development.
Shigella flexneri infection: a histopathologic study of colonic biopsies in monkeys infected with virulent and attenuated bacterial strains
Authors: KÄRnell ANDERS , Reinholt FINN P. , Katakura SHIGEHIRO , Lindberg ALF A.
Published: August 2009

Macaca fascicularis monkeys were orally infected with live virulent Shigella flexneri wild‐type strains of either serotype Y (S. flexneri SFL1), 2a (S. flexneri M4243) or lb (S. flexneri SFL27). Clinical signs of shigellosis varied from mild watery diarrhea (SFL1) to dysentery (M4243, SFL27), with a fatal outcome in two monkeys (SFL27). Colonoscopy showed slight pathologic changes in monkeys infected with SFL1, and pronounced changes in monkeys infected with SFL27. In colonic biopsies the most severe acute inflammation, with surface epithelial erosions and ulcerations, was seen after infection with SFL27, followed by SFL1, and M4243. The live S. flexneri serotype Y vaccine strain SFL114, derived from SFL1 and attenuated because of an inactivated aroD gene and hence auxotrophic for/7‐aminobenzoic acid, caused no diarrheal illness in 14 monkeys. In colonic biopsies, SFL114 only elicited a slight acute inflammatory reaction. Vaccinated monkeys were protected against clinical disease when challenged with any one of the three virulent S. flexneri wild‐type strains. Histopathologically, the acute inflammation was of less intensity than that seen in non‐vaccina ted monkeys. A good correlation between clinical signs, endoscopic findings and the degree of acute inflammation was demonstrated for monkeys vaccinated with SFL114 and challenged with either SFL1 or SFL27.
Leukaemia and anaemia in AKR/O mice I
Authors: Hellebostad MARIT , ØStbye KIRSTEN M. , Halvorsen SVERRE
Published: August 2009

AKR/O mice were used as a model for studying the pathogenesis of the anaemia accompanying leukaemia/lymphoma. The leukaemia incidence was 87%. Median age at diagnosis was 11.3 months. At diagnosis most of the mice had normal leukocyte counts. Clinically the mice divided into subgroups depending on the relative organ involvement: 1) thymoma group (n = 98), 2) spleen group (n= 144), 3) combined group (n = 27) and 4) mice with moderate organ changes (n = 216). Mice of group 1 were younger than the others, had a rapidly progressive disease, normal to elevated packed cell volume (PCV), and plasma erythropoietin (Epo) was normal or increased. Mice of group 2 were usually anaemic with high plasma Epo estimates and often elevated reticulocyte counts. Group 4 was the oldest group. Some of these mice were severely affected haematologically. Overall there was an inverse relation between PCV and plasma Epo estimate, indicating a normal Epo response to anaemia. In all groups increasing spleen size was associated with increased severity of anaemia and increased reticulocyte counts. The association between anaemia, elevated reticulocyte counts and spleen enlargement suggests haemolysis as a mechanism for anaemia, and also raises the question of compensatory spleen erythropoiesis.
Characteristics of spontaneously agglutinating Proteus mirabilis strains from bacteriuric patients
Authors: Krajewska‐Pietrasik DANUTA , Larsson PETER , Zych KRYSTYNA , Wlodarczyk JANUSZ , Gromska WIESLAWA
Published: August 2009

Out of 210 Proteus mirabilis isolates from bacteriuric patients a total of eight spontaneously agglutinating strains were found. SDS‐PAGE analysis of their lipopolysaccharides (LPS), the separation of polysaccharide fractions (PS) by gel filtration, and chemical characterization of PSs were performed. Out of the eight strains one S form and one mutant classified as Rc were detected. The remaining six strains were recognized as Ra and intermediate forms. When tested in a hematogenous infection model in mice, the P. mirabilis Rc mutant survived in kidneys for at least two weeks, while the Re mutant used as control was eliminated within 20 h after the challenge. These data indicated that strains of P. mirabilis may be pathogenic even if they express very incomplete LPS.
Ossification in nude mice. 2.
Authors: Smetana KAREL , Holub MIROSLAV , Funda DAVID
Published: August 2009

The autopodia of proximal limbs as well as the proximal growth plates of the tibia of newborn nu/ nu including super nu/nu, nu/+ and +/+mice were studied. No differences in the ossification of proximal limb autopodia (regarding the distribution of alkaline phosphatase, acid phosphatase or glycosaminoglycans) were observed in mice of genotypes studied. On the other hand, a thinner proximal tibial growth plate characterizes one‐month‐old nu/nu mice, and also the architecture and alkaline phosphatase pattern were altered. The results suggest a postnatal secondary effect of the nu/ nu genotype on skeletal development.
Primary retroperitoneal mucinous cystadenocarcinoma
Authors: JØRgensen LARS J. , Vibits HENRIK
Published: August 2009

A case of a primary retroperitoneal mucinous cystadenocarcinoma in a 38‐year‐old female is presented. The literature concerning primary retroperitoneal cystadenocarcinomas is reviewed and it is concluded that close postoperative follow‐up is necessary.
Experimental immunization with Pseudomonas aeruginosa alginate induces IgA and IgG antibody responses
Authors: Johansen HELLE KROGH , HØIby NIELS , Pedersen SVEND STENVANG
Published: August 2009

We tried experimentally to induce a specific antibody response against Pseudomonas aeruginosa locally in the airways and systemically in rats by three different routes of immunization; intragastric feeding, intratracheal inoculation or subcutaneous vaccination. Three groups of rats were immunized with live mucoid P. aeruginosa PAO 579 by intragastric feeding or with killed PAO 579 intratracheally or subcutaneously. Three other groups were immunized with purified P. aeruginosa alginate either by intragastric feeding, intratracheally or subcutaneously. At weekly intervals for four weeks animals were sacrificed and serum and bronchial fluid were obtained. The specific IgA and IgG antibody response in lavage fluid and serum was measured. Only traces of antibodies could be detected in the bronchial lavage fluids. Anti‐alginate IgA and IgG antibodies developed in all rats immunized with alginate but no antibodies against other P. aeruginosa antigens were detected. The highest IgA and IgG titer against alginate was induced by the subcutaneous immunization. IgA and IgG antibodies against other P. aeruginosa antigens developed in rats immunized with live and sonicated bacteria. The highest IgA and IgG titers were obtained after intratracheal and subcutaneous immunization with sonicated bacteria. The present work has shown that IgA and IgG antibodies develop with high specificity after immunization. The different titers obtained do not necessarily reflect different degrees of protection.
Parathyroid cell number and size in hypocalcemic young rats
Authors: Wernerson ANNIKA , Widholm SILWA MENGARELLI , Svensson OLLE , Reinholt FINN P.
Published: August 2009

Weanling rats were fed diets with normal (1%) or low (0.08% or 0.02%, respectively) Ca content for 28 days prior to sacrifice. The total volume of the parathyroids was estimated from serial sections. Volume density of secretory cells was calculated according to conventional stereological techniques, whereas cell number and cell size were estimated by the disector method. Compared with controls the animals of the experimental groups developed moderate and severe hypocalcemia and their parathyroids were enlarged with a proportional growth of parenchyma and interstitium. Related to the body weight, secretory cell volume was highest in animals with severe hypocalcemia. In the enlarged glands the size of parathyroid secretory cells was increased by 30–40%, whereas total cell number was unaltered. Thus, the increased parathyroid size was due to cell hypertrophy rather than hyperplasia.
Pancreatic islet‐cell epitope recognized by an anti‐sulphatide monoclonal antibody
Authors: Buschard KARSTEN , Josefsen KNUD , Rygaard JØRGEN , Spitalnik STEVEN L.
Published: August 2009

Insulin‐dependent (Type 1) diabetes mellitus is recognized as an autoimmune disease and islet‐cell antibody (ICA) is present in the majority of patients at diagnosis. ICA labels both beta and alpha cells and is believed to be directed against a glycolipid. In this study we examine the presence of sulphatide (3′‐sulphogalactosylceramide) or closely related structures (sulpholactosylceramide and seminolipid) in islet cells by means of a monoclonal antibody, Sulph I. Histological examination of pancreatic tissue from Lewis and BB rats, and BALB/c and NOD mice showed a pronounced labelling of the islets of Langerhans with Sulph I. No staining of the exocrine pancreatic tissue, the heart, the liver, the adrenals, the thymus, the spleen or lymph nodes was seen, but staining of some tubular cells and glomerular cells in the kidney as well as of myelin in nerve cells was found. Cytological examination of isolated Lewis islet cells and their cell subpopulations, separated using a fluorescence‐activated cell sorter (FACS), showed positive surface labelling of 97.3 ± 2.2% (SD) of the beta cells and 84.4 ± 3.0% of the non‐beta cells. Thus, the epitope on the glycolipid sulphatide or closely related structures is ‐ with the exception of neural and certain kidney tissue ‐ specifically present in islet cells. Furthermore, the staining pattern of the antibody used, Sulph I., was equivalent to that of ICA.
Two new Escherichia coli O groups: O172 from »Shiga‐like» toxin II‐producing strains (EHEC) and O173 from enteroinvasive E. coli (EIEC)
Authors: ØRSKOV IDA , WACHSMUTH I. KAYE , TAYLOR DAVID N. , ECHEVERRIA PETER , ROWE BERNARD , SAKAZAKI RIICHI , ØRSKOV FRITS
Published: August 2009

Two Escherichia coli strains were established as antigenic test strains for two new O groups, O172 and O173. The O172 strain (EHEC) which produces »Shiga‐like« toxin II (verocytotoxin 2) was isolated from a case of haemorrhagic colitis while the enteroinvasive O173 strain (EIEC) originated from a child with diarrhoea.
Study of transcription in measles virus‐infected Vero cells using cDNA probes prepared from poly(A)RNA from uninfected and infected cells
Authors: KALLAND KARL‐H. , KALVENES MAY BRITT , ØYAN ANNE MARGRETE , HAUKENES GUNNAR
Published: August 2009

From af primary plasmid cDNA library prepared from measles virus‐infected Vero cell poly(A)RNA, 435 clones selected at random were used to examine the sensitivity and specificity of cDNA probes derived from total poly(A)RNA from uninfected and infected Vero cells. The correlation between the abundance level of a particular species in the cDNA probe and the hybridization signal strength generated by the corresponding cDNA clone on a filter was reliably determined only when at least three independently prepared filters were examined. Variation in the amount of target plasmid was the most important cause of spurious signals. Variation in cDNA insert length did not disturb the signal strength within certain limits. cDNA species with abundance levels down to 0.08–0.01% were able to produce a hybridization signal above background. Unspecific cross‐hybridization was shown to define the sensitivity limit of mixed cDNA probes. Despite the many false signals present at different stages, cDNA probes provided valuable information: the cDNA probes were used to monitor relative RNA expression levels and to clone five different measles virus transcripts and 2 host cell transcripts more abundantly expressed in infected cells. The abundance levels of the measles virus nucleocapsid, phosphoprotein, matrix, fusion protein and haemagglutinin genes were 1.5%, 1.5%, 1%, 0.75% and 0.5%, respectively, of the total cDNA library.
Production of antibodies to the constant region of the mouse T‐cell‐receptor β‐chain
Authors: SCHENKEL JOHANNES , SCHÄFER HUBERT , KOCH SUSANNE , BURGER REINHARD
Published: August 2009

To generate antibodies to the constant region of the mouse T‐cell‐receptor β chain, the corresponding cDNA clone 86T5 was subcloned into the bacterial expression vector pEX2. The induced hybrid protein consisting of a 30 kDa T‐cell‐receptor segment and a β‐galactosidase carrier moiety was used for immunization. In Western blots using lysates of a T‐cell‐receptor‐positive T‐cell hybrid, the antisera obtained reacted with a 42 kDa protein under reducing and a 85 kDa protein under nonreducing conditions. The antisera did not react in binding assays with intact T lymphocytes, i.e. membrane‐associated T‐cell‐receptor protein.
Improving on BCG
Authors: STANFORD J. L.
Published: August 2009

BCG is the only vaccine for tuberculosis and leprosy known to be effective in at least some places. Unfortunately it tends to be less successful in just those areas of the developing world where a vaccine is most needed. Although molecular biology offers the prospect of alternatives, these still lie in the indefinite future, and the best use has to be made of BCG. A number of preparations are available from different manufacturers, and a vaccine should be selected with good evidence of efficacy, and a low incidence of complications. Selection of the optimal age for administering BCG should be based on factors pertaining in the area where it is to be used. The influence of contact with environmental mycobacteria, the age at which mycobacterial diseases occur, and the logistics of vaccine delivery must be taken into account. The addition of a suspension of killed Mycobacterium vaccae to BCG may increase its efficacy. Skin test data show that recognition of antigens common to all mycobacterial species and thought to be the first step in the protective immune response, is significantly enhanced by the additive. M. vaccae also contains a substance, or substances, “switching off” the tissue destructive aspect of the Koch phenomenon that is part of the immunopathology of tuberculosis. A suspension of killed M. vaccae alone can be used to enhance immune responses of persons unsuitable for BCG vaccination, such as those already tuberculin positive, and those with scars of earlier BCG vaccination.
The juxtaglomerular apparatus of the spontaneously hypertensive rat
Authors: FARAJ A. H. , MORLEY A. R. , LINDOP G. B. M.
Published: August 2009

Spontaneously hypertensive rats (SHR) are used to study the pathogenesis of essential hypertension. This study investigates the role of the renin‐angiotensin system (RAS) in the pathogenesis of hypertension in SHRs and the morphometry of the JGA by a three‐dimensional computer reconstruction program “GLOM” and electron microscopy. Systolic blood pressure (SBP) (tail cuff method) was higher in SHRs compared to controls (P<0.001). Plasma renin concentration (PRC) was lower in SHRs than in controls (P<0.001). Reconstruction of the JGA revealed granulated JG cells in the afferent and efferent arterioles and in the vascular tree away from the JGA area. Electron microscopy showed granulated JG cells in the afferent and efferent arterioles. The percentage volume of the granulated JG cells in SHR was significantly higher than in controls (P<0.01). A relationship was found between the percentage volume of granulated JG cells and the SBP in SHRs (r = 0.933, P<0.05). The wall/lumen perimeter ratio was also significantly higher in the SHRs compared to the controls (P<0.05). Low PRC in SHRs has been reported by several workers. The apparent hyperactivity of the JGA may indicate failure of renin release or an abnormal synthesis/secretion rate.
A flow cytometric method to measure the stimulated mobilization and the intracellular pool of the adhesion promoting glucoprotein Mac‐1
Authors: LUNDAHL J. , HALLDÉN G. , HED J. , JOHANSSON S. G. O.
Published: August 2009

The receptor for complement factor C3bi (Mac‐1 or CR3) belongs to a complex of leukocyte surface glucoproteins (CD11/CD18) that are essential for chemotaxis and adhesion of human polymorphonuclear leukocytes (PMN). Granulocytes can increase their surface expression of Mac‐1 upon stimulation and it is proposed that this depends on a rapid mobilization of an intracellular pool of Mac‐1. In the present study we describe a cell membrane permeabilization method that enables the detection of the intracellular pool of Mac‐1 in granulocytes by flow cytometry. The method is based on the use of the non‐ionic detergent n‐octyl‐beta‐D‐glucopyranoside (OG) to permeabilize the cell membranes of paraformaldehyde‐prefixed leukocytes. It is shown that fMLP (5 times 10‐7 M)‐treated cells expose 85% of the total detectable amount of Mac‐1 molecules on the surfaces. The method makes it possible to measure the total detectable pool, the efficiency of Mac‐1 mobilization and the in vivo expression of the receptor. This could be of value when evaluating the role of adhesion proteins in the inflammatory response.
Grouping of some clinically relevant Gram‐positive rods by automated fatty acid analysis
Authors: VON GRAEVENITZ ALEXANDER , OSTERHOUT GERARD , DICK JAMES
Published: August 2009

One hundred and ninety‐four strains of aerobically growing Gram‐positive rods of the genera Corynebacterium, Actinomyces, Arcanobacterium, Erysipelothrix, Listeria, Oerskovia, Nocardia, Rhodococcus, and of unnamed Center for Disease Control (CDC) groups were checked for cellular fatty acid profiles with the Microbial Identification System (Microbial ID, Newark, Del., USA). In order to obtain unified data usable for the clinical laboratory, 24 or 48 h sheep blood agar cultures were used. It was thought that grouping and perhaps identification could be aided by this approach. With the aid of numerical analysis, four groups (two consisting of two subgroups each) were established. The discriminatory ability of the scheme, however, was only 77.2%, indicating that grouping of an unknown isolate could be done with some accuracy, but that speciation would still require biochemical testing.
Bone marrow stroma in idiopathic myelofibrosis and other haematological diseases.
Authors: LISSE IDA , HASSELBALCH HANS , JUNKER PETER
Published: August 2009

Bone marrow stroma was investigated immunohistochemically in 31 patients with haematological diseases, mainly idiopathic myelofibrosis (n = 8) and related chronic myeloproliferative disorders (n = 14). The bone marrow from patients with idiopathic myelofibrosis and some CML patients showed marked staining reactions with antibodies against type III procollagen (pN collagen), type IV collagen, fragment PI of laminin and factor VIII. Patients with osteomyelosclerosis had particularly increased collagen content, including both newly deposited type III collagen (pN collagen) and mature collagen fibres. As in normal bone marrow, argyrophilic fibres and type III collagen displayed a close co‐distribution, which was also demonstrated for type IV collagen and laminin. While normal bone marrow sinusoids had discontinuous basement membranes, fibrosing bone marrow was characterized by endothelial cell proliferation and capillarization, with the development of continuous sheets of basement membrane material beneath endothelial cells.
Frequency of SV40‐specific cytotoxic T‐lymphocyte precursors in two SV40 T‐antigen transgenic mouse lines
Authors: JURETIC ANTONIO , KNOWLES BARBARA B.
Published: August 2009

Cytotoxic T‐lymphocyte precursors (CTL‐P) from control C57BL/6 mice and from mice of two simian virus 40 (SV40) T‐antigen transgenic lines, 427 and 419, specifically nonresponsive and responsive, respectively, to SV40 T antigen, were quantitated by limiting dilution (LD) after immunization with SV40. CTL‐P frequencies for the SV40 T antigen‐responsive 419 line transgenic mice were within the range established in C57BL/6 mice, whereas no CTL‐P could be demonstrated for the SV40 T antigen‐tolerant 427 line mice. These results suggest that deletion or anergy of SV40 T antigen‐responsive clones underlies the specific profound tolerance of 427 line mice.
Redistribution of myeloperoxidase and elastase in human neutrophil granulocytes
Authors: NÄSSBERGER L.
Published: August 2009

Both myeloperoxidase (MPO) and elastase were found to be redistributed in ethanol‐and Bouin‐fixed human neutrophil granulocytes. Air‐dried and formalin‐fixed cells, stained for MPO, appeared with a cytoplasmic fluorescence and unstained nuclei. In elastase air‐dried cells a faint perinuclear staining was also detected. Otherwise the redistribution pattern was very similar for MPO and elastase. It was found that both elastase and MPO were most efficiently extracted with Bouin's fixative. A discontinuous fluorescence was seen especially for MPO in ethanol‐fixed cells. Nuclear membrane staining was verified with laser scan microscopy. This technique also visualized a scattered fluorescence in the nuclear matrix for MPO.
Formation of osteoclasts and osteoblast‐like cells in long‐term human bone marrow cultures
Authors: KASSEM M. , MOSEKILDE Li. , RUNGBY J. , MOSEKILDE Le. , MELSEN F. , ERIKSEN E. F.
Published: August 2009

A culture system for human osteoclasts is described. It is based on incubation of mononuclear marrow cells for three weeks in the presence of 1,25 dihydroxyvitamin D3. The resulting cells exhibited all the classic characteristics of osteoclasts. They formed a true syncytium with up to 25 nuclei per cell. The cells revealed a strong positive reaction for tartrate‐resistant acid phosphatase and a ruffled border. When these cells were cultured on 100 μm thick human dentine slices for three weeks, typical resorption pits were formed. In the culture, alkaline phosphatase‐positive osteoblast‐like cells were formed in the vicinity of osteoclasts. This culture system permits studies of human osteoclasts and their interaction with osteoblast cell precursors in the human bone marrow.
Formation of osteoblast‐like cells from human mononuclear bone marrow cultures
Authors: KASSEM M. , RISTELI L. , MOSEKILDE L. , MELSEN F. , ERIKSEN E. F.
Published: August 2009

Osteoblast‐like cells are commonly found in the vicinity of osteoclasts formed in long‐term human bone marrow cultures, and they are believed to be derived from osteogenic cell precursors belonging to the stromal cell system. This paper describes a new culture method for human osteoblasts from the adherent cell population of long‐term human mononuclear bone marrow cultures. The cells obtained exhibited all the classic characteristics of osteoblasts. They contained high intracellular concentrations of alkaline phosphatase and they secreted the osteoblast‐specific marker bone Gla protein. Collagen production was mainly (95–98%) procollagen type I propeptide and only minute quantities of procollagen type III propeptide were detectable by radioimmunoassay in the conditioned medium. After eight weeks the cells formed a mineralized matrix on exposure to β‐glycerophosphate and ascorbic acid. This system provides a model for the study of osteoblast differentiation in vitro and may form the basis for the use of defined media in bone cell cultures due to the presence of high concentrations of osteoblast precursors.
Effect of interferon treatment on expression of gC and gE glycoproteins in herpes simplex virus‐infected cells
Authors: HOLBERG‐PETERSEN MONA , BÖHMER ELLEN , NAMORK ELLEN , DEGRE MIKLOS
Published: August 2009

The effect of interferon treatment on the herpes simplex virus type 1 (HSV‐1)‐specific glycoproteins gC and gE in homologous and heterologous cells has been investigated. In human embryonic fibroblastic cells, human leukocyte interferon inhibited virus multiplication and expression of the HSV‐1‐specific glycoproteins gC and gE on the cell surface in a dose‐dependent manner. In heterologous baby hamster kidney cells, the human interferon had no effect on virus multiplication. However, the surface expression of the HSV‐1‐specific glycoproteins was reduced, as shown by erythrocyte rosette formation, by attachment of monodisperse polystyrene particles coated with antibodies and by immunogold scanning electron microscopy.
Rapid identification of Legionella pneumophila zinc metalloprotease using chromogenic detection
Authors: McINTYRE MICHAEL , QUINN FREDERICK D. , FIELDS PATRICIA I. , BERDAL BJORN P.
Published: August 2009

Strains of Legionella spp. produce extracellular proteases that can be detected using synthetic chromogenic peptides. Chromogenic tri‐ and tetrapeptides show a high degree of sensitivity, specificity and reagent stability when linked to para‐nitroaniline (pNA). For example, SucOMe‐Arg‐Pro‐Tyr · pNa (S‐2586) is specifically hydrolysed by proteases of Legionella pneumophila and some other Legionella species. A paper disc method to sample protease directly from agar plates has been used to evaluate chromogenic peptides as reagents for diagnostic purposes. Strains of Legionella spp., Pseudomonas spp. and Enterobacteriaceae were examined, together with a recombinant Escherichia coli strain containing the cloned 38 kDa zinc metalloprotease from L. pneumophila. S‐2586 was hydrolysed by 282 out of 283 L. pneumophila strains, and by the recombinant E. coli. Two of the six strains representing other Legionella species, and 22 of the 50 strains from the Pseudomonas group were also positive. No reaction was seen with any of the Enterobacteriaceae strains. Although there was functional homology between proteases from several bacterial groups, the high prevalence of S 2586‐hydrolysing proteases within L. pneumophila indicates a potential usefulness for phenotypic identification.
Autoantibodies in patients with IgA and IgG2 deficiencies
Authors: JIMÉNEZ ADOLFO , ALVAREZ‐DOFORNO RITA , RODRÍGUEZ M. CRUZ GARCÍA , FERREIRA ANTONIO , LÓPEZ‐TRASCASA MARGARITA , FONTAN GUMERSINDO
Published: August 2009

Patients with primary immunodeficiencies have a high incidence of autoantibodies, mainly of no clinical significance. It has recently been suggested that patients with a combined IgA‐IgG2 deficiency have more autoantibodies than those patients with isolated deficiencies. We have studied 42 patients with selective IgA deficiency, nine with isolated IgG2 deficiency and 13 with combined IgA‐IgG2 deficiency, and have found that the combined IgA‐IgG2 deficiency has no influence on autoantibody prevalence, except for anti‐IgA antibodies. The presence of chronic respiratory infections (a clinical feature commonly associated with both selective IgA and IgG2 deficiencies) is unrelated to the prevalence of autoantibodies. The most frequent autoantibodies found are anti‐IgA and anti‐cardiolipin. Most of the autoantibodies have been found to be devoid of actual clinical significance. Only three patients had overt autoimmune disease.
Fibronectin in bronchoalveolar lavage fluid and plasma of dogs with acute inflammation of the lungs
Authors: NAGY B. , KATONA É. , ERDEI J. , KARMAZSIN L. , FACHET J.
Published: August 2009

Bronchoalveolar inflammation, which was generated in dogs by Broncho‐Vaxom instilled into the right lower lobe, was characterized first of all by an increased influx of macrophages. In this nonpurulent acute‐subacute inflammatory reaction, the lavage fibronectin decreased rapidly three hours after the incubation and then a marked gradual elevation was observed, which persisted throughout the whole two‐week process, while plasma fibronectin concentrations were not altered significantly. Changes in the levels of lavage fibronectin may be an important sign for the control of the inflammatory reaction activity in the lungs.
Cytokeratin intermediate filament pattern in uterine cervical biopsies
Authors: NIELSEN L. N. , HØRDING U. , DAUGAARD S. , RASMUSSEN L. P. , JOHANNESEN C. L. , NORRILD B.
Published: August 2009

Twenty‐seven uterine cervical biopsies with histological diagnoses ranging from normal through dysplasia to invasive carcinoma were analysed for cytokeratin pattern in two‐dimensional gel electrophoresis. No direct correlation between histological diagnosis and cytokeratin pattern was observed.
The effects of blockade of interleukin 2 receptors and interleukin 4 receptors on cytokine production
Authors: LAI K. N. , LEUNG J. C. K. , LI P. K. T. , LARCHE M. , RITTER M. A.
Published: August 2009

The understanding of the regulation of cytokine production and T‐cell proliferation by other cytokines is far from complete. We have examined the supernatant concentration of different cytokines in cultured peripheral blood lymphocytes (PBL) incubated with monoclonal antibodies against the interleukin‐2 receptor (IL‐2R) or interleukin‐4 receptor (IL‐4R) complex in order to analyse the importance of these receptors in the production of other cytokines. The PBL cultures were stimulated with phytohaemagglutinin and phorbol myristate acetate. The addition of anti‐Tac to PBL cultures resulted in increased free IL‐2 due to blockade of IL‐2R. MR6 (a monoclonal antibody against IL‐4R) did not affect the supernatant concentration of IL‐2 in cultured PBL. Blockade of IL‐2R or IL‐4R both resulted in increased IL‐4 in supernatant. The increase in supernatant concentration of IL‐4 in PBL cultured with MR6 was due to a direct interaction with the IL‐4R complex but the increase in IL‐4 with anti‐Tac was due to the accumulation of free IL‐4 as an effective blockade of IL‐2R prevented the full expression of IL‐4R. Neither anti‐Tac nor MR6 affected the IL‐6 production by activated PBL, suggesting the existence of an IL‐2‐ and IL‐4‐independent pathway. Blockade of IL‐2R or IL‐4R both resulted in a reduction of interferon‐gamma (IFN‐gamma) production, indicating that both endogenously generated IL‐2 and IL‐4 are important mediators of IFN‐gamma induction in PBL cultures stimulated with T‐cell mitogens.
Turpentine‐ and fibrin‐induced pleuritis in nude mice
Authors: FAARUP P. , FAURSCHOU P. , GRUNNET N.
Published: August 2009

Turpentine was injected into the right pleural cavity of nude immune‐incompetent mice, causing a temporary irritative exudative pleuritis. A transient occurrence of so‐called rheumatoid arthritis cells was observed in the pleural fluid together with parallel characteristic biochemical changes. In similar experiments in nude mice, however, immunization followed by intrapleural application of bovine fibrin showed irritative “dry” pleuritis without the presence of rheumatoid arthritis cells. This is in contrast with previous results from similar experiments done using normal mice. The conclusion from the present experiments is that in nude immune‐incompetent mice only the non‐immunological, turpentine‐induced pleuritis will generate cellular and biochemical changes typical of the rheumatoid disease in patients, while the fibrin‐induced pleuritis fails to show similar changes. This suggests that the rheumatoid‐like pleural changes described in the present experiments in nude mice have a non‐immunological basis.
Macrophages in trauma‐induced myositis ossificans
Authors: ARO HANNU T. , VILJANTO JOUKO , AHO HEIKKI J. , MICHELSSON JARL‐ERIK
Published: August 2009

The basic cellular mechanisms of different forms of myositis ossificans are poorly known. In the current experiment the nature of the early (24 168 h) inflammatory cell reaction preceding traumainduced myositis ossificans was studied. New bone formation was induced in the vastus intermedius region of the rabbit quadriceps muscle by means of immobilization and daily passive mobilization. Before the start of treatment, a cell harvesting device (viscose cellulose sponge in a silastic tube) was implanted in the region of interest. The opposite intermedius muscle and a standardized surgical skin wound served as the control sites. The results showed a significantly prolonged invasion of macrophages into the ossifying intermedius muscle as compared with the control intermedius muscle. It is hypothesized that microinjury and subsequent muscle necrosis cause the invasion of macrophages, and these cells respond to the conditions of the traumatized muscle under passive mobilization by releasing osteogenic growth factors.
Genome fingerprinting as a typing method used on polyagglutinable Pseudomonas aeruginosa isolates from cystic fibrosis patients
Authors: OJENIYI B. , HØIBY N. , ROSDAHL V. T.
Published: August 2009

Phenotypical changes occur in the surface of Pseudomonas aeruginosa during the chronic lung infection of cystic fibrosis patients. It is difficult with the classical typing methods, such as serotyping, phage typing and pyocin typing, to decide if a patient has been colonized with a new strain or whether it is the same strain which has reappeared, for instance after chemotherapy in the lungs. This investigation was carried out to evaluate genome fingerprinting as a typing method and to see how it correlated with classical methods and with DNA probe typing. Forty Pseudomonas aeruginosa isolates, 34 polyagglutinable and six monoagglutinable, from 14 cystic fibrosis patients were analysed using genome fingerprinting. The bacterial chromosomes were digested with the restriction endonucleases Dra 1 and Xbal, and separated by field inversion gel electrophoresis. The results were compared with those of a previous work (Ojeniyi et al. 1990) concerning typing with a DNA probe, serotyping using both polyclonal and monoclonal sera, phage typing, pyocin typing and reverse phage typing. The results of genome fingerprinting and DNA probe typing showed the best correlation, followed by pyocin typing. The correlation between the results of genome typing and the other typing methods was low. The discriminatory effect of genome fingerprinting was higher than that of DNA probe typing, and genome fingerprinting was found to be the best single method for epidemiological investigations of polyagglutinable isolates from cystic fibrosis patients.
Mechanism of aminoglycoside resistance in Danish Staphvlococcus aureus strains during the years 1979–1987
Authors: MADSEN OLE RINTEK , LORENTZEN JAN SØBO , FRIMODT‐MØLLER NIELS , MORTENSEN IDA , ROSDAHL VIBEKE THAMDRUP
Published: August 2009

A total of 49 gentamicin‐resistant strains of Staphylococcus aureus isolated from blood (n = 26) or from other sites (n = 23) during the years 1979 to 1987 were evaluated for the presence of aminoglycoside‐inactivating enzymes on the basis of minimum inhibitory concentrations measured by agar dilution as well as inhibition zone diameters determined by disc diffusion. Enzymatic activity was caused by AAC (6′)III/APH (2″) in 45 strains, and by AAC (6′)III/APH (2″)+APH (3′) in four strains. No changes in the distribution of enzymatic activity were observed during the eight years.
Epithelial changes following bone marrow transplantation: cytological atypia and epithelial endocrine cells
Authors: DESPERBASQUES MARTA , DHILLON AMAR , HAMID QUATAYBA
Published: August 2009

This report describes the histological and immunocytochemical findings in gastrointestinal biopsies and skin of a patient with chronic granulocytic leukaemia which progressed to blastic transformation, who was then treated with chemotherapy and total body irradiation followed by allogeneic bone marrow transplantation. The gut showed endocrine cells in the lamina propria and these had an immunophenotype similar to the glandular epithelium at the sites studied (stomach, duodenum and rectum), supporting the idea that the endocrine cells of the gut are more resistant to the effect of radiation, chemotherapy and graft‐versus‐host disease (GVHD) than are other cell types in the epithelium, and that lamina proprial endocrine cells are epithelially derived in this situation. Epidermis and gut epithelium also showed marked atypia due to the conditioning regimen and GVHD, and this case illustrates the possibility of misdiagnosis of carcinoma in this increasingly common situation. An additional, unusual feature of this case was the presence of a B‐cell UCHL1+ve lymphoma of the colon at autopsy, 122 days post‐transplantation.
Multivariate analyses of cellular fatty acids and carbohydrates of 1:2:1 and 2:4:2 spirochetes
Authors: BRONDZ ILIA , FIEHN NILS‐ERIK , OLSEN INGAR , SJÖSTRÖM MICHAEL
Published: August 2009

Delimitation of small‐sized spirochetes of the oral cavity can be difficult. The endoflagella pattern has long served as the main criterion for taxonomic distinction. For approved species, e.g. Treponema denticola, several endoflagella patterns are observed, which indicates that this criterion is inadequate. The present study with GC and GC‐MS used fatty acids and carbohydrates of whole‐cell methanoly‐sates to distinguish 1:2:1 and 2:4:2 subgingival spirochetes. Thirteen fatty acids: C12:0, C13:0, C14:0, Ciso‐14:0, C2‐OH‐14:0, C15:0, Cantc‐15:0, C16:0, Ciso‐16:0, C16:1, C18:0, C18:1, and C18:2, and three carbohydrates: rhamnose, glucose, and glucosamine, were detected. The carbohydrate contents did not differ between the two groups. While 1:2:1 spirochetes contained Ciso‐14:0 and Cantc‐15:0 acid, 2:4:2 spirochetes did not. Also multivariate analyses of quantitative fatty acid data distinguished between these groups.