bluuebunny/crossref_metadata_2025 huggingface dataset:
Are Intestinal bacteria involved in the etiology of rheumatoid arthritis?
Authors: HAZENBERG MAARTEN P. , KLASENM INA S. , KOOL JEANETTE , EMBDEN JOHANNEKE G.H. RUSELER‐VAN , SEVERIJNEN ANTON J.
Published: August 2009

Observations in bowel‐related joint diseases give support to this hypothesis. In Crohn's disease and ulcerative colitis, the bowel wall inflammation is complicated in about 20% of the patients by joint inflammation. Bowel infection bySalmonella, ShigellaandYersiniacan provoke joint inflammation and supports an etiological link between bowel bacteria and arthritis. The arthropathic properties of the most abundant group of intestinal bacteria, i.e. the obligate anaerobic bacteria, were studied in an animal model. Cell wall fragments (CWF), with peptidoglycan as the major component, from someEubacteriumandBifidobacteriumspecies induced a severe chronic polyarthritis in Lewis rats after a single intraperitoneal injection.Eubacteriumwas found in numbers of 108‐109per gram in stools of healthy subjects and rheumatoid arthritis (RA) patients. CWF of isolated strains ofE. aerofacienswere arthropathic. Soluble peptidoglycan polysaccharide complexes (PG‐PS) originating from the obligate anaerobic flora were purified from human intestinal contents. PG‐PS from ileostomy fluid that proved to be less processed by intestinal enzymes induced chronic arthritis in rats after a single administration in oil in the base of the tail. It was concluded that the human intestinal bowel contains soluble bacterial cell wall products that are arthropathic in an animal model. Peptidoglycan (PG) or its subunits was reported to be present in mammalian tissues. Immunohistochemical studies from our group showed the presence of intestinal PG‐PS in sections of normal rat spleen. Bacterial cell wall or PG‐induced joint inflammation in rats is proven to be absolutely dependent on functional T cells. T‐cell lines were isolated from the lymph nodes of rats with anE. aerofaciensCWF arthritis. A helper T‐cell line B13 wasin vivoarthritogenic in knee or ankle joints upon intravenous injection in rats and proliferatedin vitroon syngeneic spleen cells alone, but was additionally stimulated by intestinal PG‐PS andE. aerofaciensCWF. It was postulated that the arthritogenic T cells that seem to be autoreactive are, in fact, recognizing bacterial PG‐PS on antigen‐presenting cells (APC). It is generally accepted that RA is a T‐cell‐dependent process and that therefore the reaction is directed at small peptides bound by the major histocompatibility complex of APC. The only peptides present in arthritis inducing intestinal PG‐PS and in CWF are PG peptides interlinking the sugar chains. We feel that the immunoreaction against PG peptides plays a pivotal role in experimental and human arthritis of an unknown etiology.
Two types of receptors for human plasminogen on group G streptococci
Authors: ULLBERG MANS , KARLSSON INGA , WIMAN BJÖRN , KRONVALL GÖRAN
Published: August 2009

To investigate the nature of plasminogen binding to streptococci, strains selected for high reactivity with human plasminogen were examined for binding pattern against a panel of plasminogen fragments. The strains included human isolates of groups A, C and G as well as bovine isolates of group G. All strains reacted substantially with the plasminogen fragment kringle 1–3. Using the miniplasminogen fragment (kringle 5 and the B chain) a small but reproducible uptake was detected for human group G strains but not for group A or C strains. The group G strains of bovine origin on the other hand demonstrated high uptake of miniplasminogen, suggesting the possibility of an alternative plasminogen receptor for this species. This interpretation was supported by blocking experiments with the lysine analogue EACA where low concentrations (1 mM) completely blocked plasminogen binding to human streptococci, whereas a 100‐fold higher concentration was needed for bovine group G strains. Scatchard plots with human isolates resulted in straight lines and Kd values were generally in the range of 20–80 nM. The number of receptors was estimated to be 45,000 for a selected group A strain and about 10,000 for the selected group C and G strains. Scatchard analysis with bovine group G isolates on the other hand revealed a two phase interaction, supporting the assumption of two different receptor structures on these strains. Kd for the first phase was estimated to be about 20 nM (10,000–20,000 receptors per bacterium), which was similar to the human strains, whereas the second phase was in the range of 400–500 nM (50,000 and 150,000 receptors per bacterium with two selected strains). Scatchard plots with the miniplasminogen fragment as ligand mimicked the phase two reaction with plasminogen, supporting the concept that this reaction represents a new and not previously described receptor. Both the receptor reacting with the kringle 1–3 portion and the one reacting with the miniplasminogen portion bound plasmin and plasminogen with similar affinity.
Local IgA and IgG response to intratracheal immunization with Pseudomonas aeruginosa antigens
Authors: JOHANSEN HELLE KROGH , HØIBY NIELS
Published: August 2009

We have experimentally immunized rats intratracheally with sonicated Pseudomonas aeruginosa to develop a high antibody response systemically and locally. The systemic antibodies were measured by a standard enzyme‐linked immunosorbent assay (ELISA) on serum samples, whereas the local antibodies were measured on eluates from paper discs containing saliva. High levels of IgA antibodies were elicited in saliva whereas only traces of IgG antibodies were detected; the opposite results were found in serum. The saliva disc method permits serial measurements from the same animal and therefore offers the possibility to follow post‐immunization antibody responses.
The glomerular peripolar cell ‐ an immunohistochemical study
Authors: GARDINER D. S. , LINDOP G. B. M.
Published: August 2009

The peripolar cell is a recently described glomerular epithelial cell which may be a new secretory component of the juxtaglomerular apparatus (JGA). The purposes of this study were to identify immunohistochemical markers specific for peripolar cells and to elucidate the composition of their cytoplasmic granules. We examined tissue from normal human and sheep kidneys with a variety of monoclonal antibodies and antisera. A minority of peripolar cells showed immunoreactivity for vimentin but no other intermediate filament. Neuroendocrine markers were negative. The granules of peripolar cells contain a variety of plasma proteins with the exception of IgM. It is likely that peripolar cells absorb plasma proteins from the glomerular filtrate. The reason for this is uncertain; however, their unique position at the vascular pole of normal kidneys suggests a specialized function.
Enzymuria in neonates receiving continuous intravenous infusion of gentamicin
Authors: COLDING HANNE , BRYGGE KIRSTEN , BRENDSTRUP LISBETH , BENTZON MICHAEL W. , ANDERSEN GUNNAR E.
Published: August 2009

Urinary excretion of the tubular enzymes NAG and AAP was investigated during gentamicin treatment of 105 newborn infants. The values found for NAG and AAP show a significant positive correlation. The urinary excretion of NAG was on the average 92% higher during gentamicin treatment as compared with non‐treatment periods in the same newborn infant (33 infants). The same tendency applied to AAP. Newborn infants receiving continuous intravenous infusion of gentamicin were not found to be at greater risk of nephrotoxicity than those receiving intermittent gentamicin treatment, using NAG and AAP as an index of nephrotoxicity. The changes in NAG and AAP within treatment periods were studied. During gentamicin treatment an insignificant average increase in the urinary excretion of NAG occurred, whereas a significant decrease was found during non‐treatment periods. A significant negative correlation was found between urinary excretion of NAG and birth weight/gestational age. The long‐term effect of the higher excretion of NAG and AAP in newborn and adult patients during aminoglycoside treatment is unknown.
Effect of interferon‐α on measles virus replication in human peripheral blood mononuclear cells
Authors: LEOPARDI ROSARIO , HYYPIÄ TIMO , VAINIONPÄÄ RAIJA
Published: August 2009

We analyzed the effect of exogenous human leukocyte interferon (IFN)‐α on measles virus (MV) replication in human peripheral blood mononuclear cells (PBMC). The release of infectious virus was progressively reduced by increasing concentrations of IFN‐α, and blocked with an IFN‐α concentration of 1000 U/ml. In order to detect a possible target of this inhibitory effect, viral transcription and translation events were analyzed. The synthesis of MV mRNAs was reduced, but not blocked, in the presence of IFN‐α. However, this effect was not specific on the viral RNAs, but due to a general inhibition of RNA synthesis in IFN‐treated PBMC. The expression of viral polypeptides was also inhibited in a dose‐dependent manner by exogenous IFN‐a, but a low level of protein synthesis was detected by both Western blotting and immunofluorescence techniques, even with the maximum amount of IFN‐α used (1000 U/ml). These findings account for a partial maintenance of the viral replicative cycle, even when the production of infectious virus is blocked. Moreover, the effect of IFN‐α is not specifically targeted on the virus macromolecular synthesis.
Enterotoxin and toxic shock syndrome toxin‐one production by staphylococci isolated from mastitis in sheep
Authors: ORDEN JOSE ANTONIO , CID DOLORES , BLANCO MARIA EMILIA , QUITERIA JOSE ANTONIO RUIZ SANTA , GÓMEZ‐LUCIA ESPERANZA , DE LA FUENTE RICARDO
Published: August 2009

From 160 staphylococci isolated from ovine mastitis, 125 were identified as coagulase‐positive staphylococci (CPS) and 35 as coagulase‐negative staphylococci (CNS). Of these, 108 (87.8%) S. aureus produced at least one of the staphylococcal enterotoxins (SE) described. However, no CNS was found to be enterotoxigenic. Enterotoxin C (SEC) was the type most frequently produced. TSST‐1 was shown to be produced by 91 (74.0%) of S. aureus, almost invariably in combination with SEC. Three CNS strains were also found to produce TSST‐1 (two strains of S. xylosus and one strain of S. epidermidis).;
Relation between enzyme‐linked immunosorbent assay and radioimmunoassay for detection of antibodies to the capsular polysaccharide of Haemophilus influenzae type b
Authors: KRISTENSEN KIM , BENTZON MICHAEL WEIS
Published: August 2009

The measurement of antibodies to the capsular polysaccharide (PRP) of Haemophilus influenzae type b (Hib) is important because vaccines inducing such antibodies are now available. We developed and evaluated an enzyme‐linked immunosorbent assay (ELISA) for detection of these antibodies based on direct coating of the plates with tyraminated PRP. The assay fulfilled the requirements for parallel line assays; it was sensitive, specific, and reproducible with a coefficient of variation between days of 19%. Results from the ELISA were compared with results from radioimmunoassay and a correlation coefficient of 0.93 was found. Results obtained by the two methods were proportional and the relation was independent of the antibody level. The relation between them was also unaffected by the contribution of different antibody isotypes, indicating that these were measured to the same extent by both methods. ELISA employing direct coating of the plates with tyraminated PRP represents a useful alternative for detection of antibodies when studying immunogenicity of Hib vaccines.
Serum resistance in Escherichia coli strains causing acute pyelonephritis and bacteraemia
Authors: JACOBSON STEFAN H. , ÖSTENSON CLAES‐GÖRAN , TULLUS KJELL , BRAUNER ANNELIE
Published: August 2009

The capacity of Escherichia coli to resist the bactericidal action of serum was examined in 367 clinical isolates obtained from children with acute pyelonephritis (n = 57), adults with acute pyelonephritis (n = 55), non‐diabetic patients with bacteraemia (n = 101), diabetic patients with bacteraemia (n = 65) and from the faecal flora of healthy controls (n = 89). The incidence of serum‐resistant E. coli strains was significantly higher in pyelonephritogenic strains from children and adults (93% and 82%) as compared to faecal control strains (57%, p < 0.001 and p< 0.005 respectively). Strains causing bacteraemia in non‐diabetic and diabetic patients were more often serum resistant (72% and 80%) as compared to control strains (p < 0.05 and p < 0.001 respectively). The frequency of serum‐sensitive strains was similar in diabetic patients with decreased renal fuction or proteinuria compared to those with normal renal function. There were no significant correlations between serum resistance of E. coli and expression of P fimbriae, type I fimbriae or mannose‐resistant haemagglutination, cell surface hydrophobic properties, production of aerobactin, haemolysin or cytotoxic necrotizing factor in 53 pyelonephritogenic strains from adult patients.
Fine structure of neutrophils from turpentine‐induced pleuritis in mice, simulating rheumatoid arthritis cells
Authors: NIELSEN MORTEN H. , FAURSCHOU PETER , FAARUP POUL
Published: August 2009

Pleural effusions were made by intrapleural turpentine installation in mice. The fine structure of inflammatory cells from the effusions was normal except for lipid inclusions. The same type of inclusion was previously found in neutrophils from pleural effusions in patients with tuberculous infection, rheumatoid disease, or carcinomatosis. The lipid inclusions observed in neutrophils from an irritative turpentine‐induced pleurisy should be considered as “fatty change”, and are structurally similar to the rheumatoid arthritis cells seen in patients with different diseases.
Plasmid profiles as an epidemiological marker in Salmonella enterica serovar berta infections
Authors: OLSEN J. E. , SØRENSEN M. , BROWN D. J. , GAARSLEV K. , BISGAARD M.
Published: August 2009

Plasmid profiles of 674 Salmonella berta strains isolated in the period 1985–1989 from 674 cases of human infections were recorded. Five hundred and eighty‐nine of the strains examined (87.4%) contained plasmids and the mean number of plasmids was 1.8 per strain. Sixteen different plasmid sizes were recognized, 13 of which had also been reported from S. berta of poultry origin. The plasmids formed 90 different profiles. Forty‐five of these profiles, comprising 90.7% of isolates, had previously been recorded among poultry strains. Only 12 human isolates (1.8%) carried plasmids which were not demonstrated among isolates obtained from broilers. The prevalences of the 20 most commonly demonstrated profiles were not significantly different among isolates obtained from humans and broilers. The average difference between the prevalences of individual plasmid profiles was 2.1, with a standard deviation of 4.2. Based on the results, indications are that broilers represent the main reservoir for human S. berta infections in Denmark.
Correlation between coxsackie B1 virus replication and enhanced invasiveness of Shigella flexneri
Authors: MODALSLI KRISTIN , BUKHOLM GEIR , MIKALSEN SVEIN OLE , DEGRÉ MIKLOS
Published: August 2009

Coxsackie Bl virus infection enhances the susceptibility of cultured HEp‐2 cells to Shigella flexneri invasiveness. This can be reproduced partially with UV‐inactivated virus, particularly the effect observed shortly after viral inoculation. The following phases of viral multiplication were correlated with the enhancing effect: uncoating of viral particles, synthesis of viral RNA and proteins, and assembly of newly produced virus particles. Uncoating of virus particles was completed within 60 min. This process was not correlated with the development of the early effect on invasiveness. Intact virus capsids seem to be necessary to enhance bacterial invasiveness in the early phase of virus infection. Separated capsid proteins had no effect either when applied to the cell surface or when microinjected into the cells. Virus protein synthesis was not required for the virus effect on bacterial invasiveness in the early infection phase, but it seems to be necessary in the late phase.
The histology at various stages of acute rejection in concordant xenogeneic heart transplantation
Authors: NIELSEN BJARNE , STEINBRÜCHEL DANIEL A. , LÆNKHOLM ANNE VIBEKE , KEMP EJVIND
Published: August 2009

In an attempt to describe early morphologic changes in heterotopic xenogeneic heart transplantation a sequential study was performed in a hamster‐to‐rat model. Mild morphologic changes observed after four to six h were characterized by slight interstitial edema and focal myocyte damage with fragmentation and loss of myofibrillar elements. No lymphocytic infiltration appeared. Moderate morphologic changes observed after 12–24 h were characterized by moderate interstitial edema, and the appearance of mild hemorrhage and scattered extravasated neutrophilic granulocytes. The myocardium had more widespread areas with myocyte damage, sometimes with small foci of necrotic cells and adjacent neutrophilic granulocytes and macrophages. Vascular changes with perivascular edema and swelling of the endothelium were seen and a few neutrophilic granulocytes could be found in the vessel walls. No lymphocytic infiltration appeared. Severe morphologic changes observed after 44–48 h or at the time of complete rejection were characterized by severe interstitial hemorrhage, appearance of widespread necrosis and marked vascular changes with development of leukocytoclastic‐like vasculitis, possibly with thrombosis. Only a few lymphocytes appeared. The findings were essentially different from those observed in allogeneic heart transplantations, where classical first‐set allograft rejection was seen. In normal donor hearts and syngeneic transplanted hearts used as controls, no significant morphologic changes were demonstrated. On the basis of this study we consider xenogeneic acute rejection to be primarily of the humoral type.
The progression of cisplatin‐induced tubulointerstitial nephropathy in rats
Authors: MARCUSSEN NIELS , JACOBSEN N. O.
Published: August 2009

We have previously shown that administration of cisplatin to rats leads to chronic nephropathy with many atubular glomeruli. Using unbiased Stereological and histochemical methods our purpose was to investigate the progression of renal structural changes, specifically the events preceding and following the formation of atubular glomeruli in rats after increasing total doses of cisplatin. After administration of cisplatin (2 mg/kg body weight i.p. once weekly) the animals were sacrificed 10 days, three, six, 10 and 20 weeks later. After 4 mg/kg the distal part of most proximal S3 segments appeared necrotic with a negative reaction for enzymes characteristic of brush border and mitochondria. After larger doses, atrophy (and possibly necrosis) was observed in clusters of cortical proximal and distal nephron segments corresponding to an increasing amount of non‐functioning nephrons. Hypertrophy/hyperplasia of remaining uninjured nephrons and collecting ducts was observed. The Stereological investigations showed an increasing number of atubular glomeruli with open capillaries or glomeruli connected to atrophic tubules with the total dose of cisplatin (30% and 40%, respectively, after 20 mg/kg). The percentage of glomeruli with damaged tubules was significantly higher in the juxtamedullary cortex compared with the superficial cortex. Sclerotic glomeruli were not observed. The findings suggest that a toxic destruction of most S3 segments leads to atrophy or disappearance of the corresponding nephrons with formation of atubular glomeruli, and that the remaining hypertrophic nephrons are connected to the few surviving or regenerated S3 segments.
Tumours in Iceland 15. Ependymoma
Authors: BIRGISSON SIGURBJÖRN , BLÖNDAL HANNES , BJÖRNSSON JOHANNES , OLAFSDOTTIR KRISTRUN
Published: August 2009

Thirteen ependymomas reported to the Icelandic Cancer Registry during a 32‐year period (1955–1986) were histologically reviewed and reclassified according to the WHO Histological Typing of Tumours of the Central Nervous System. The annual incidence rate of ependymoma was 0.20/100.000. Clinical observations and data on biological behaviour and immunohistochemistry are presented. Four tumours were supratentorial, six infratentorial and three intraspinal. There were ten males and three females with a mean age of 32 years (range 2.5–68). The mean postoperative survival of nine surgically treated patients was 35.5 months. Histologically, eight tumours were classical ependymomas, three anaplastic and two myxopapillary. Of 11 tumours stained for GFAP, nine were positive. Nine of 10 tumours tested were positive for vimentin, five for NSE and four for S‐100. None of the 10 tumours showed reactivity with AFP, CEA, chromogranin, desmin, factor VIII, keratin or neurofilament.
Alloxan cytotoxicity involves lysosomal damage
Authors: ZHANG HONG , ZDOLSEK JOHANN M. , BRUNK ULF T.
Published: August 2009

The cytotoxic effects of alloxan are not understood in any great detail, although they are considered to involve reactions mediated by oxygen‐derived free radicals. These reactive species may form extra‐or intracellularly following alloxan reduction, and result in cell damage through a number of complex interactions with a variety of macromolecules. The purpose of the present study was to elucidate further the early intracellular effects of alloxan on a model system of macrophage‐like cells in culture. Addition of alloxan (15 mM), without reducing agents, to the medium surrounding the cells (phosphate‐buffered saline, PBS, 37°C, pH 7.4) resulted in rapid lysosomal damage (disappearance of the proton gradient over the membrane) followed by severe cellular degeneration (swelling and blebbing) and 50% cell death (trypan blue dye exclusion test) within fifty min. Cells pretreated with the γ‐glutamyl cysteine synthetase‐inhibiting agent BSO, to decrease levels of intracellular glutathione, showed enhanced sensitivity to alloxan. The results are interpreted as indicating the cytotoxicity to result from intracellular formation of superoxide radicals, hydrogen peroxide and hydroxyl radicals, the latter within secondary lysosomes containing trace amounts of reactive iron (inducing Fenton reactions). The ensuing lysosomal membrane damage may result in leakage of lysosomal hydrolases and further cellular degeneration.
Extracellular reduction of alloxan results in oxygen radical‐mediated attack on plasma and lysosomal membranes
Authors: ZHANG HONG , GAO GUIXIAN , BRUNK ULF T.
Published: August 2009

Alloxan participation in extracellular redox processes results in the formation of the reactive oxygen species (ROS) superoxide anions (O2−), hydroxyl radical (OH·) and hydrogen peroxide (H2O2), causing cell damage through a number of complex interactions probably involving several different cellular structures. These involve the plasma membrane, and we have recently presented evidence for lysosomal interference. The present study elucidates the early (within 15 min) events in a model system of macrophage‐like cells (J‐774) in culture. Addition of 2 mM alloxan and 1 mM cysteine to the medium surrounding the cells (phosphate‐buffered saline, PBS, 37°C, pH 7.4) resulted in rapid lysosomal membrane damage with disappearance of the proton gradient as visualized by acridine orange relocalization, as well as plasma membrane alterations leading to increased leakage of fluorescein after fluorescein diacetate staining. These events were later (> 30 min) followed by cellular degeneration in the form of blebbing. Mitochondrial damage (rhodamine 123 relocalization) was a late event. Cells pretreated with desferrioxamine (Des) and superoxide dismutate (SOD) or Des, SOD and catalase (CAT) to induce partial (H2O2formation only) or almost full protection (no ROS formation) showed about the same reactions as when cells were exposed to alloxan and cysteine without scavengers (O2−, H2O2and OH· formation) or with PBS only, respectively. The results are interpreted as indicating that the cytotoxicity is a consequence mainly of H2O2involvement and probably of lysosomal influx of H2O2with ensuing OH· formation within secondary lysosomes containing trace amounts of reactive iron. It is suggested that the resultant lysosomal membrane damage is followed by leakage of lysosomal hydrolases and ensuing cellular degeneration.
Mucosal immunity to Pseudomonas aemginosa alginate in cystic fibrosis
Authors: PEDERSEN SVEND STENVANG , MØLLER HENRIK , ESPERSEN FRANK , SØRENSEN CHRISTIAN HJORTH , JENSEN TIM , HØIBY NIELS
Published: August 2009

Patients with cystic fibrosis commonly acquire chronic pulmonary infection with alginate‐producing Pseudomonas aemginosa. The infection remains localized at the mucosal surfaces of the airways. Using enzyme‐linked immunosorbent assays immunoglobulin concentrations and titers of specific antibodies to purified P. aeruginosa alginate and to P. aemginosa sonicated antigens were measured in tears, saliva, sputum and serum. CF patients had significantly higher concentrations of IgG, IgA and SIgA in serum and saliva than controls. They also had significantly higher levels of specific antibodies to alginate and sonicated antigen in secretions and serum. Local production of IgA, IgG and IgM antibodies to P. aeruginosa was demonstrated. Only a minor proportion of specific IgA antibodies were present as secretory IgA in tears, saliva and sputum. The ratio of alginate‐specific SIgA to specific monomeric IgA in sputum was significantly lower than the similar ratio in saliva, whereas the same ratio for specific P. aeruginosa sonicate antigens was found in saliva and sputum.
Immunohistochemical characterization of an amphicrine mucinous islet‐cell carcinoma of the pancreas
Authors: LAINE VELI J. O. , EKFORS TAUNO O. , GULLICHSEN RISTO , NEVALAINEN TIMO J.
Published: August 2009

Immunohistochemical characteristics of a mucinous islet‐cell carcinoma of the pancreas are described. The tumour presented with jaundice in a 59‐year‐old male. It consisted of polygonal atypical cells forming a reticular pattern, and invaded the common bile duct. In DNA flow cytometry, the tumour cells showed a clear‐cut aneuploid peak. Intercellular mucin was abundant. A panel of antisera and monoclonal markers was applied in the immunohistochemical analysis. In addition to general epithelial and endocrine markers, the tumour cells showed a focal positive immunoreaction with anti‐glucagon, anti‐insulin, anti‐vasoactive intestinal polypeptide, anti‐pancreatic secretory trypsin inhibitor and antiphospholipase A2 antigen. At the ultrastructural level, mucous and neuroendocrine granules were demonstrated in the same tumour cells.
Immunohistochemical studies of β‐hexosaminidase in neoplastic and normal tissues with monoclonal antibodies
Authors: ELSAFI MOHAMED ELHASSAN , ISAKSSON ANDERS , STENRAM UNNE
Published: August 2009

A total of 87 human specimens with 10 histological types of primary neoplasm were studied immunohistochemically with monoclonal antibodies specific for β‐hexosaminidase (Hex). High levels of Hex were found in malignant neoplasms of the skin, cervix, colorectum and in benign as well as neoplastic plasma cells, while no activity was detected in normal epidermis, normal colorectal epithelium or benign naevi. The strongest immunohistochemical reaction was revealed in tumor cells of malignant melanoma. Adenomas and adenocarcinomas of the colorectum showed high levels of Hex with a basal pattern of immunoreactivity more frequent in the tumor cells of adenocarcinomas than adenomas. Fibroblasts and macrophages in the tumors often disclosed immunoreactivity. In most of the sections (including those from plasma cell neoplasms), 7E4 antibody showed low immunoreactivity compared to 2E3, except for non‐neoplastic plasma cells, which were as a rule positive with 7E4 and largely negative with 2E3 antibody. This result probably indicated different isoenzymes in benign and neoplastic plasma cells.
Origin and evolution of the vertebrate immune system
Authors: DU PASQUIER LOUIS
Published: August 2009

The immune system is a complex evolutionary unit and it would be simplistic to conclude that the immune systems of all primitive vertebrates are primitive. Because of the large number of elements involved, many evolutionary events must have taken place, some of them neutral, some of them selected, to constitute the systems that we are looking at towards the end of the 20th century. All these systems have perhaps evolved beyond the apparent evolutionary state of the species in which they are found. They have been modulated by factors linked not only to the internal evolution of their elementary genes, but also by coevolution with factors in the internal environment, such as cellular constraints, metabolism, mode of reproduction and progeny size. It seems that good inventions are long lasting, which is the reason why some elements of the invertebrate immune system can be found with similar functions in vertebrates (defensins). It is also the reason why Ig domains have been exploited in so many ways, whether for the immune system or not. Again, they had an evolution of their own. The comparative study of the immune systems carried out on the occasion of this phylogenetic survey shows a world particularly dynamic and diverse. The comparisons between the solutions chosen by the various phyla of the animal kingdom, or closer to us by the various classes of vertebrates, allow us to distinguish the essential features of the immune system. From this viewpoint, this approach is not only of phylogenetic interest, but also has an applied aspect. Increasing our knowledge in this area could help suggest solutions to clinicians when they are faced with deficiencies and abnormalities in the immune system of man.
Superoxide anion release from blood monocytes and alveolar macrophages in patients with diffuse lung fibrosis
Authors: SHERSON D. , NIELSEN H. , FREDERIKSEN J. , MILMAN N. , STRUVE‐CHRISTENSEN E. , PETERSEN B. NUCHEL
Published: August 2009

Superoxide anion release (O2−) after stimulation with phorbol myristate acetate was measured in alveolar macrophages (AM) obtained by bronchoalveolar lavage and in blood monocytes from 47 patients with diffuse interstitial lung disease: idiopathic pulmonary fibrosis (N = 15), hypersensitivity pneumonitis (N = 7), pneumoconiosis (N = 6) and sarcoidosis (N= 19). Differential cell counts demonstrated a lymphocyte predominance in patients with hypersensitivity pneumonitis (HP) and sarcoidosis while the other groups had neutrophil predominance. No correlation between O2− activity in alveolar macrophages (AM) or blood monocytes (BM) compared to lung function (VC and diffusing capacity) could be demonstrated. Smoking pneumoconiotics had significantly decreased BM Of release (1.25 ± 0.25 (SEM) nmol/min/106 cells) and significantly increased AM/BM O2− ratios (2.04±0.26) compared to smokers with idiopathic pulmonary fibrosis (IPF) who had the following mean values: BM O2− release = 2.58±0.25 and AM/BM O2− ratio=0.86 ± 0.23. When matched for sex and smoking, a significantly increased AM/BM O2− ratio was seen among patients with HP (2.19 ± 0.98) in comparison with patients who had sarcoidosis (0.40±0.18). Patients with either HP or pneumoconiosis had generally elevated AM O2− release and reduced BM O2− release. These results suggest that environmentally related interstial lung disorders (HP and pneumoconiosis) may be associated with elevated AM O2− release relative to BM O2− release in comparison to non‐environmentally related disorders (IPF or sarcoidosis).
Proliferative myositis and fasciitis
Authors: LUNDGREN LARS , KINDBLOM LARS‐GUNNAR , WILLEMS JAN , FALKMER URSULA , ANGERVALL LENNART
Published: August 2009

Two cases of proliferative myositis, four cases of proliferative fasciitis and one mixed form of proliferative myositis and fasciitis have been analyzed in terms of cell differentiation and DNA content. Light microscopically, the lesions were characterized by a mixture of proliferating spindle‐shaped cells and uni‐, bi‐ or occasionally multinucleated ganglion cell‐like cells. The spindle cells showed ultrastructural features and immunohistochemical properties, including an immunoreactivity for smooth muscle‐specific actin, indicative of a myofibroblastic differentiation. The ganglion cell‐like cells displayed some resemblance to active osteoblasts ultrastructurally and differed immunohistochemically from the spindle cells by being non‐immunoreactive for smooth muscle‐specific actin. None of the two cell types showed immunoreactivity for desmin, myoglobin or factor VIII RAG. It is suggested that the two cell types represent different lines of cell differentiation. The cytologic features in smears, as seen in two cases of proliferative fasciitis and one case of proliferative myositis, are considered to be characteristic of these lesions and to permit the diagnosis to be made by fine‐needle aspiration. In two of the cases, the lesion was diagnosed only cytologically and thereafter disappeared spontaneously within a month. Cytometric DNA measurements, using two different image analysis systems on Feulgen‐stained sections and smears, revealed a “diploid” spindle‐shaped cell population with a variable proportion of cells with scattered DNA values. The ganglion cell‐like cells differed from the spindle cells by having a broad DNA peak in the diploid region and additional peaks in the tetraploid region, as well as a higher proportion of cells with scattered DNA values compared with those of the spindle‐shaped cells. The results of the quantitative DNA analysis are well in keeping with the benign and proliferative nature of these lesions. However, with the technique used here, quantitative DNA analysis, does not distinguish these pseudosarcomatous fibrous lesions from diploid and tetraploid soft tissue sarcomas.
Ultrastructure of human osteoblasts and associated matrix in culture
Authors: KASSEM MOUSTAPHA , RUNGBY JØRGEN , MOSEKILDE LEIF , ERIKSEN ERIK FINK
Published: August 2009

The ultrastructure, as visualized by transmission electron microscopy, of cells obtained from human bone explants and subsequently cultured is described along with the electron microscopic appearance of the associated intercellular matrix. The cells were characterized as osteoblasts on the basis of immunohistochemical, enzymatic, and functional criteria. Although the osteoblasts could be cultured in standard culture media and always appeared singly, not forming syncytia, the cultures were eventually confluent and formed multilayers. The cells were fusiform or cuboidal with diameters ranging between 10–15 μm. The cytoplasm was characterized by numerous large mitochondria, and especially by a very prominent RER. The intercellular matrix was woven with collagen fibres surrounding large numbers of matrix vesicles. In areas with matrix vesicles, evidence for osteoblast activity, i.e. mineralization related to matrix vesicles, could be observed after incubation with β‐glycerophosphate. In conclusion, we provide evidence that human osteoblasts cultured in vitro synthesize collagen and produce a matrix with vesicles capable of initiating mineralization processes.
Interleukins increase surface ganglioside expression of pancreatic islet cells in vitro
Authors: KJAER T. W. , RYGAARD J. , BENDTZEN K. , JOSEFSEN K. , BOCK T. , BUSCHARD K.
Published: August 2009

This experiment was conducted in order to investigate whether expression of gangliosides on islet cell surface in vitro is influenced by cytokines, especially interleukin 1. Islets from adult Lewis rats were incubated with different concentrations of recombinant‐derived human cytokines. Following dispase treatment, the single cells were labeled with monoclonal antiganglioside antibodies A2B5 or R2D6, and conjugate. Both are directed against β cells; A2B5 is recognized to bind specifically to pancreatic islet cells, while R2D6 is shown to bind no other pancreatic cells than β cells. Surface labeling was evaluated in blind trials using a fluorescence microscope and a fluorescence‐activated cell sorter (FACS). A2B5 staining demonstrated a significantly higher number of labeled cells after incubation with interleukin la (14.9%±2.8; p<0.005), interleukin 1β (23.2%±4.2; p<0.0005) or TNFα (16.1%± 4.0; p = 0.005) compared to endotoxin controls (4.1%±1.1). Interleukin 1β (9.5%β1.5; p<0.005) showed a significantly increased number of R2D6‐stained cells (control: 2.3%±1.3). A similar but not significant effect was seen with interleukin 1α and TNFα. Interleukin 6 had no effect on the antigen expression. The intensity of labeling was elevated among interleukin 1β‐incubated cells compared to control samples. Thus, treatment of islets with different cytokines, especially interleukin 1β, increases surface antigen expression. We suggest that this mechanism of action in vitro may be of importance for the putative diabetogenic effect of interleukin 1.
Enhancement of normal polymorphonuclear cells respiratory burst in ascitic fluid by fibronectin
Authors: LEBRUN LÉA , PELLETIER GILLES , BRIANTAIS MARIE‐JOSÉ , BRIVET MICHÈLE , KHELLAF AMINA , GALANAUD PIERRE
Published: August 2009

The chemiluminescence (CL) response of normal peripheral blood polymorphonuclear cells (PMN) in ascitic fluids (cirrhotic = 32; malignant = 17) was studied independently of the ascitic fluid complement activity. CL response and fibronectin levels were higher in malignant ascitic fluid than in cirrhotic ascitic fluid (p< 0.001). Addition of pure fibronectin or malignant ascitic fluids to cirrhotic ascitic fluids increased the CL response of normal PMN. These findings suggest that the susceptibility of cirrhotic patients to spontaneous bacterial peritonitis (SBP) is a multifactorial defect involving factors distinct from low C3 levels. Fibronectin is an important factor in the promotion of the respiratory burst of normal PMN stimulated by opsonized zymosan or PMA in ascitic fluid. Our results suggest that low levels of ascitic fluid fibronectin could partly explain the high susceptibility of cirrhotic patients to spontaneous bacterial peritonitis.
Disseminated histoplasmosis in a badger (Meles meles) in Denmark
Authors: JENSEN H. E. , BLOCH B. , HENRIKSEN P. , DIETZ H. H. , SCHØNHEYDER H. , KAUFMAN L.
Published: August 2009

We report the first case of disseminated histoplasmosis in an animal in Scandinavia. Yeast cells compatible with those of Histoplasma capsulatum var. capsulatum were found in the skin, liver, spleen, a kidney, and a lymph node of a wild badger (Meles meles). The diagnosis was confirmed by electron microscopy and immunofluorescence staining of the yeast cells in tissue sections.
HSV‐IgA serum antibodies in cervical intraepithelial neoplasia and invasive cancer patients, and in their spouses: a case control study
Authors: GUPTA MADAN M. , JAIN RENU , PARASHARI ADITYA , SINGH V. , SATYANARAYANA L.
Published: August 2009

Class‐specific IgG and IgA antibodies to HSV were assayed in women with CIN (76), invasive cancer (52) (histological diagnosis) and age‐matched controls (119), employing HSV‐2‐infected HEp‐2 cells as antigen during IFA assay. We observed an elevated geometric mean titre (GMT) of serum antibody (IgG five‐to eight‐fold and IgA four‐to five‐fold) for the entire spectrum of cervical lesions, as compared to controls. The odds of finding HSV‐IgA antibodies were highest with CIN III (OR = 22.0), followed by invasive carcinoma, and CIN I & II (OR = 9.5 and 5.2), respectively. Furthermore, the investigations with respect to married couples (husbands and wives) who volunteered to participate in this study (33 cases and 47 control group) also indicated relatively high antibody titres and increased frequency of HSV sero positivity amongst husbands of cases as compared to their wives, as well as the control group males and females. The contribution of HSV infection in women and/or their husbands to the risk of developing abnormal cervical lesions was analysed after adjusting for the same in respective counterparts. It was observed that the risk was increased 14‐fold with HSV‐IgA positivity of women, and that HSV‐IgA positivity of husbands (male partners) further increased the risk 16‐fold. This preliminary observation shows the importance of serum HSV‐IgA antibodies as a risk indicator in cervical precancer and cancer lesions in women without a history of recent genital herpes lesions. The serum HSV‐IgA may also be taken as an indicator of “high risk” males.
Antigenic change in a human IgG4‐specific CH3 epitope upon binding of a monoclonal antibody against a neighboring IgG4‐specific epitope
Authors: ROLSTAD ANNA KRISTIN , MICHAELSEN TERJE E. , KOLBERG JAN
Published: August 2009

Two sets of monoclonal antibodies (mAbs) probably reacting with two different epitopes in the CH3 domain of the human IgG4 molecule were studied. We observed that the commercially available mAb HP 6011 inhibited the antigen binding of the three mutually inhibitable mAbs, 40‐A2, 41‐E8 and 43‐F11 (40‐series), made by us. However, the 40‐series mAbs, including those with similar affinity such as mAb HP6011, were not able to inhibit mAb HP 6011. When the 40‐series mAbs were preincubated with IgG4, the mAb HP 6011 could partially displace these antibodies. This one‐way inhibition indicates that upon binding mAb HP 6011 changes the antigenic structure of the IgG4 molecule by disrupting the epitope for the 40‐series mAbs. A steric hindrance of this epitope by mAb HP 6011 is more unlikely, since the small Fab fragment of mAb HP 6011 also inhibited the reaction of the 40‐series mAbs.
Growth patterns of pulmonary metastases and primary tumours from five murine fibrosarcoma cell clones
Authors: FALKVOLL KNUT H. , GRIMSTAD IVAR AMUND
Published: August 2009

The growth patterns, including the size, shape and regional preferences, of lung metastases from five murine fibrosarcoma cell clones were studied. Spontaneous metastases developed from tumours formed by subcutaneous inoculation of the cell clones. Lung colonies (experimental metastases) were established by i.v. injection of cells. The numbers of both spontaneously and experimentally formed subpleural lung metastases were counted through a stereomicroscope. The fraction of colonies that was located subpleurally was determined in histological sections of lungs. The growth kinetics of clonally derived primary tumours, and the number of spontaneous and experimental lung metastases, differed greatly between certain cell clones. The number of spontaneous lung metastases was correlated with the maximum size of primary tumours. No close correlation was observed between the size of the primary tumours and the size of experimental metastases. There were differences between the cell clones in the shape and regional preferences of their lung deposits. The subpleural colonies were generally larger than the intrapulmonary ones. Thus, both the regional distribution and the growth pattern of lung deposits differed between the clones.
Rotavirus infection in Tanzania: a virological, epidemiological and clinical study among young children
Authors: SAM N. E. , HAUKENES G. , SZILVAY A. M. , MHALU F.
Published: August 2009

Rotavirus infection in the Dar es Salaam area of Tanzania was studied in 99 hospitalized children with acute diarrhoea and 99 hospitalized non‐diarrhoea referents matched for sex and age. Of the diarrhoea cases 43.4% had rotavirus in the stools as opposed to 15.2% of the referents. The high carrier rate among the referents represents a serious risk of nosocomial transmission. More referents than cases had serum IgG antibodies to rotavirus, 52.5% and 35.4%, respectively (P<0.02), while there was no correlation with serum IgM and IgA or faecal IgA antibodies. The latex agglutination test had a sensitivity comparable to that of electron microscopy (100%) and a specificity of 93.8%. The Slidex test appeared to be superior to the Rotalex test in that it gives very few false‐positive reactions. The SDS‐PAGE patterns of 11 RNA segments were compatible with the presence of group A strains with considerable heterogeneity among the strains. Symptoms and signs and some environmental data were recorded. None of them was clearly associated with rotavirus infection among the diarrhoea cases. It is concluded that rotavirus is a major cause of acute infectious diarrhoea in Tanzania.
Cancer‐associated changes in glycosylation of fibronectin
Authors: MANDEL ULLA , THERKILDSEN MARIANNE HAMILTON , REIBEL JESPER , SWEENEY BETH , MATSUURA HIDEMITSU , HAKOMORI SEN‐ITIROH , DABELSTEEN ERIK , CLAUSEN HENRIK
Published: August 2009

The extracellular matrix adhesion molecule fibronectin exhibits different isoforms derived by alternative splicing as well as recently demonstrated variation in O‐glycosylation. Although fibronectin is widely distributed in normal tissues, the individual isoforms have been found to show restricted tissue distribution and association with malignancies. The monoclonal antibody FDC‐6 defines a cancer‐associated de novo glycosylation of a specific threonine residue in the C‐terminal region of the fibronectin molecule termed oncofetal fibronectin. Here we report an immunohistological study of oral squamous cell carcinomas (n = 33), premalignant lesions (n = 15), and normal oral mucosa (n = 10) using the FDC‐6 antibody. A selective expression of the oncofetal fibronectin epitope was demonstrated in close relation to the invading carcinoma, whereas no staining was observed in premalignant lesions without epithelial dysplasia, or in normal epithelium. Furthermore, we attempted to identify additional carbohydrate‐related epitopes distinguishing fibronectin of human hepatoma cell line HUH‐7 from plasma fibronectin. No novel epitopes were identified, as all generated monoclonal antibodies lacking reactivity with plasma fibronectin showed the same specificity as FDC‐6. Previous studies have indicated that the de novo glycosylation is induced by a novel transferase activity only found in fetal and carcinoma cell lines, placenta and hepatoma tissues. Here we provide further evidence that a purified UDP‐GalNAc:peptide N‐acetylgalactosaminyltransferase from normal bovine thymus and human placentae is incapable of utilizing the hexapeptide VTHPGY as a substrate. The results demonstrate that oncofetal fibronectin is highly associated with malignancy, and appears to be induced by expression of a unique glycosyltransferase or modification of the specificity of the normally expressed transferase.
Estimation of the total number of mast cells in the human umbilical cord
Authors: DAMSGAARD T. M. ENGBERG , NIELSEN B. WINDELBORG , SØRENSEN F. B. , HENRIQUES U. , SCHIØTZ P. O.
Published: August 2009

The aim of the present study was to estimate the total number of mast cells in the human umbilical cord. Using 50 μm‐thick paraffin sections, made from a systematic random sample of umbilical cord, the total number of mast cells per cord was estimated using a combination of the optical disector and fractionated sampling. The mast cell of the human umbilical cord was found in Wharton's jelly, most frequently in close proximity to the three blood vessels. No consistent pattern of variation in mast cell numbers from the fetal end of the umbilical cord towards the placenta was seen. The total number of mast cells found in the umbilical cord was 5,200,000 (median), range 2,800,000–16,800,000 (n = 7), that is 156,000 mast cells per gram umbilical cord (median), range 48,000–267,000. Thus, the umbilical cord constitutes an adequate source of mast cells for further investigation of these cells in the newborn, e.g. for describing their functional and secretory characteristics and possible clinical relevance in relation to the development of allergic, inflammatory and immunological diseases in infancy and childhood.