bluuebunny/crossref_metadata_2025 huggingface dataset:
A new Danish Listeria monocytogenes phage typing system
Authors: GERNER‐SMIDT P. , ROSDAHL VIBEKE THAMDRUP , FREDERIKSEN W.
Published: August 2009

A phage typing system was developed for Listeria monocytogenes. Phages were released from clinical and from food strains of L. monocytogenes by mitomycin C induction. The system consists of two subsystems, one of which is used to type L. monocytogenes serotype 1 strains, containing 12 phages, and a second which is used to type serotype 4 strains, containing 14 other phages. For the serotype 1 subsystem, the reproducibility was ≥90%, the typability 92%, and the discriminatory power, as judged by the discriminatory index, 80%. The corresponding figures for the serotype 4 subsystem were: reproducibility ≥94%, typability 87%, and discriminatory index 87%. The performance of the whole typing system is sufficient for it to be used for screening purposes.
Comparison of genome fingerprinting with conventional typing methods used on Pseudomonas aeruginosa isolates from cystic fibrosis patients
Authors: OJENIYI B. , PETERSEN U. STEEN , HØIBY N.
Published: August 2009

Two hundred Pseudomonas aeruginosa serial isolates from 61 cystic fibrosis patients were examined using restriction fragment length polymorphism in connection with pulsed field gel electrophoresis. A comparison was made of results obtained by genome fingerprinting and by conventional typing methods. It was possible to subdivide the majority of the genome types with the conventional typing methods, indicating the likelihood of bacterial phenotypes occurring in the lung of the cystic fibrosis patient. Two clusters of strains were observed among the 200 P. aeruginosa isolates from the 61 patients. Strains belonging to one cluster were present in 26 (42.6%) of the 61 patients. Strains belonging to the other cluster were present in 11 (18.0%) of the 61 patients. The occurrence of these clusters indicates that cross‐infection has taken place among CF patients attending the Danish Cystic Fibrosis Centre. Conventional typing methods are based on the presence of specific bacterial surface structures. Therefore, conventional typing methods may sometimes lead to wrong classification of isolates from cystic fibrosis patients, especially if applied alone. A combination of two to six methods decreased the reproducibility of the typing results. The best combination was genome fingerprinting and phage typing, which yielded a reproducibility of 82.5%. Each time a method is added, the reproducibility decreases by an average of 14.4%.
T cells in reactive arthritis
Authors: HERMANN ELISABETH
Published: August 2009

T cells appear to play a major role in the development, maintenance and also resolution of reactive arthritis (ReA). Recent advances in understanding the processes involved in T cell activation now allow us to examine the peripheral blood and synovial fluid T cell responses to given “arthritogenic” microorganisms in terms of antigen specificity, epitope identification, cytokine secretion patterns, HLA restriction and the role of different T cell subsets in ReA. Peripheral blood bulk proliferation and limiting dilution studies provide evidence that the peripheral T cell response against arthritis‐associated gram‐negative bacteria is decreased in patients developing immunological sequelae such as ReA after gastrointestinal infection. Using clonal analysis of synovial fluid CD4+ T cells it has been shown that a polyclonal rather than an oligoclonal response to a variety of bacterial antigens is induced at the site of synovitis and that these CD4+ T cells produce a Thl‐type of cytokine. 65 kD heat shock protein may represent one of the possible linkages of anti‐infectious and autoimmune reactions. Furthermore, a spectrum of killer cells is present in the synovial fluid of patients with ReA. This spectrum of cytotoxic T cells includes antigen‐specific, class I‐restricted αβ‐TCR+CD8+ lymphocytes, antigen‐specific, apparently non‐MHC‐restricted αβ‐TCR+CD8+ lymphocytes and γδ‐TCR+ cells with broad cytolytic activity directed against bacteria‐infected target cells. HLA‐B27‐restricted Yersinia‐ or Salmonella‐specific synovial fluid CD8+ T cells may provide the missing link between genetic disposition (HLA‐B27) and extra‐articular infection with arthritogenic bacteria in these patients.
Expression of vitronectin and fibronectin binding by Candida albicans yeast cells
Authors: JAKAB ERZSÉBET , PAULSSON MARIANNE , ASCENCIO F. , LJUNGH ÅSA
Published: August 2009

Expression of binding to vitronectin (Vn or S‐protein) and fibronectin (Fn) was common among clinical isolates of Candida albicans. Growth at 37°C enhanced expression of both Vn and Fn binding. Some strains expressed higher binding after growth in liquid media and others after growth on solid media. Most strains expressed higher cell surface hydrophobicity after growth on agar media. Vn binding was less influenced by expression of cell surface hydrophobicity than Fn binding. Vn binding to yeast cells was optimal around pH 4 and Fn binding around pH 6. Binding to soluble Vn was inhibited by unlabelled Vn and to a lesser extent by Fn. Fn binding to the same C. albicans strain was inhibited by unlabelled Fn, Vn, fibrinogen and to some extent collagens. C. albicans strain 3248 expressed specific high binding of Vn, and high binding of Fn. Binding of both proteins was sensitive to heat and protease treatment, but in different ways. Vn binding differed significantly from the earlier reported Fn binding and may represent a novel type of tissue adherence.
Chronic Pseudomonas aeruginosa lung infection in normal and athymic rats
Authors: JOHANSEN HELLE KROGH , ESPERSEN FRANK , PEDERSEN SVEND STENVANG , HOUGEN HANS PETTER , RYGAARD JØRGEN , HØIBY NIELS
Published: August 2009

We have compared a chronic lung infection with Pseudomonas aeruginosa embedded in alginate beads in normal and athymic rats with an acute infection with free live P. aeruginosa bacteria. The following parameters were observed and described: mortality, macroscopic and microscopic pathologic changes, and antibody responses. The rats challenged with P. aeruginosa alginate beads experienced a generally more severe lung pathology and the antibody responses were more homogeneous with less dispersion as compared to the rats having free live P. aeruginosa bacteria. In general, manifestations were more severe in the athymic rats compared to the normal rats. It is, however, notable that the athymic rats developed similar microscopic lung manifestations as the normal rats when given a large number of P. aeruginosa in the beads, with dense accumulation of neutrophil granulocytes and microcolonies comparable to the lesions seen in cystic fibrosis (CF) patients. Early transitory IgM titers were demonstrated in both normal and athymic rats. Whilst athymic rats produced much lower IgG titers than the normal rats, presumably due to the absence of CD4+ cells, higher primary IgA titers were achieved. These two models in normal and athymic rats of the chronic lung infection resembling that seen in CF lungs make it possible to study the influence of the various components of the specific and nonspecific defence systems on the course of the chronic P. aeruginosa lung infection and to evaluate the effect of various immunization schedules and immunomodulatory drugs.
Postmortem findings and prenatal diagnosis of Zellweger syndrome
Authors: LINDHARD A. , GRÆM N. , SKOVBY F. , JEPPESEN D.
Published: August 2009

The postmortem findings of siderosis, renal cortical cysts, pachygyria, cortical heterotopia of the brain and cerebellar hypoplasia in a seven‐week‐old infant with craniofacial dysmorphism and hypotonia prompted the diagnosis of Zellweger syndrome. This was confirmed by analysis of very‐long‐chain fatty acids in blood spots from filter paper, collected in the neonatal period, and allowed first trimester diagnosis in the subsequent pregnancies.
Effect of a human IgG preparation rich in antibodies to a wide range of lipopolysaccharides on gram‐negative bacterial sepsis in burned mice
Authors: FOMSGAARD ANDERS , HOLDER IAN ALAN
Published: August 2009

A human intravenous IgG preparation (Anti‐LPS IgG) rich in antibodies to different lipopolysaccharides (LPS) and a normal human intravenous IgG (NIgG) were investigated for their ability to confer passive immunity. Both preparations were given at the time of infection (prophylaxis) or during sepsis (therapy) to burned mice with lethal infection induced by various clinically relevant gram‐negative bacteria. When given at the time of infection both IgG preparations (5 mg/mouse) inhibited lethality induced by some bacteria (Pseudomonas aeruginosa serogroup G and B), but not others (Serratia marcescens, Klebsiella pneumonia, Proteus mirabilis), indicating a protection by strain‐specific antibodies. However, no significant protection was seen when mice were treated during sepsis. The range of specific antibody titers to the whole live bacteria and heat‐killed (LPS‐preserved) bacteria in the NIgG paralleled that of Anti‐LPS IgG; however, the magnitude of the antibody titers did not accurately reflect the protective capacity in vivo. Thus, the exact specificity of the protective antibodies is still unknown. The protective effect of both IgG preparations was dose‐dependent; at low IgG doses (0.5 mg/mouse) better protection was obtained with Anti‐LPS IgG, whilst at higher doses (> 1 mg/mouse) both preparations exhibited identical effects. Low doses of either IgG preparation in combination with subtherapeutic doses of piperacillin significantly enhanced early survival (day 2 for NIgG and day 2 + 3 for Anti‐LPS IgG) against P. aeruginosa, but the protective effect waned thereafter. We conclude that a strain‐specific antibacterial effect in a compromised mouse infection model can be obtained by early passive immunization with human IgG from large plasma pools. It is suggested that Anti‐LPS IgG or NIgG may be of benefit in some cases of gram‐negative sepsis when administered as prophylaxis together with proper antibiotic treatment.
Simple mucin‐type carbohydrate antigens in pleomorphic adenomas
Authors: THERKILDSEN MARIANNE HAMILTON , MANDEL ULLA , CHRISTENSEN MARIA , CLAUSEN HENRIK , DABELSTEEN ERIK
Published: August 2009

Simple mucin‐type carbohydrate structures, T, Tn and sialosyl‐Tn, are regarded as general markers of carcinomas in several epithelial tissues as a result of incomplete synthesis with precursor accumulation. The structures have a very limited distribution in normal tissues and secretions, including saliva and salivary glands. The expression of simple mucin‐type carbohydrate structures and ABH(O) variants was studied in paraffin‐embedded and frozen tissue sections from 37 pleomorphic adenomas with associated normal parotid tissue, using immunohistology and a panel of MAbs with well‐defined specificity for T, Tn, sialosyl‐Tn, and blood group H and A variants hereof. The immature Tn and sialosyl‐Tn antigen structures were expressed in the epithelial ductular structures of the tumors, whereas they were almost absent from normal parotid tissue, indicating aberrant glycosylation with accumulation of precursor structures. Furthermore, the tumors showed loss of A antigen. The prognostic significance of these results is discussed. The modified myoepithelial cells in periductular and solid areas expressed T and sialosyl‐T antigens, similar to normal myoepithelial cells and basal cells. Thus these modified MEC seem to have retained their normal simple mucin‐type glycosylation pattern, suggesting that T antigen may be used as a marker of MEC in salivary gland tumors.
Effect of Legionella pneumophila sonicate on killing of Listeria monocytogenes by human polymorphonuclear neutrophils and monocytes
Authors: RECHNITZER CATHERINE , BANGSBORG JETTE M. , SHAND GEOFFREY H.
Published: August 2009

Legionella pneumophila shares with other intracellular pathogens the ability to resist intracellular killing within phagocytes. An increasing number of cellular components of L. pneumophila are proposed as pathogenic factors of the organism. At the site of infection, the phagocytic cells will be exposed to bacterial components, either expressed on the surface of the organisms or released in the environment upon cell lysis. In this study, we have investigated the effect of water‐soluble bacterial components present in L. pneumophila sonicate on the phagocytosis and bactericidal activity of human polymorphonuclear neutrophils and monocytes. Preincubation of neutrophils with L. pneumophila sonicate did not affect phagocytosis of L. monocytogenes, whereas Listeria killing was significantly inhibited at sonicate concentrations of 1 and 2 mg/ml. The phenol phase of a phenol‐water extraction, containing most of the lipopolysaccharide (LPS), had no inhibitory effect on the listericidal activity of neutrophils. Killing of Listeria by monocytes was inhibited in a similar manner. The inhibitory activity was mainly recovered in the sonicate fraction above 100 kDa, suggesting that components organized in larger molecular complexes are most likely to represent the inhibitory factors. The inhibitory activity of L. pneumophila sonic extract appears to be related to inhibition of killing mechanisms since uptake of Listeria was not affected by the sonicate. Our observations indicate that as Legionella infection progresses, bacterial components liberated by cell lysis could exert a detrimental effect on the antimicrobial function of phagocytes, stressing the importance of early treatment of Legionnaires' disease to reduce bacterial numbers in the infected tissues.
Cerebrovascular disease at autopsy and in the death statistics for the county of Hordaland, Norway
Authors: KARWINSKI B.
Published: August 2009

Between 1975 and 1984 a decrease in deaths from cerebrovascular disorders was found both at autopsy and in the official mortality statistics. In the autopsy material (662 cases) the decrease was seen mainly in patients dying of cerebral hemorrhage. Concordance between autopsy and mortality statistics was best in the 30 to 79‐year age group. Discordance rose with age, being highest in those above 80 years. This was mainly due to use of the diagnosis “undetermined vascular lesion” given as the clinical cause of death. The diagnosis was not upheld at autopsy. In such cases the presence of a cerebral lesion was rarely confirmed at autopsy. The decrease in acute cerebrovascular deaths was not accompanied by an increase in the number of patients in the category “stroke with sequelae”, assessed as a fall in number. Thus mortality statistics can be directly misleading in geriatric patients; and the autopsy frequency in this age group is too low to give a satisfactory picture of events.
Late changes in the normal mouse bladder after irradiation alone or in combination with cis‐DDP or cyclophosphamide, assessed by stereological analysis
Authors: LUNDBECK FINN , NIELSEN KARSTEN , STEWART FIONA
Published: August 2009

Stereological analysis has been performed to investigate the late changes in the normal mouse bladder one year after radiation alone (10–25 Gy) or radiation (10–15 Gy) combined with cyclophosphamide (CTX), 100 mg/kg, or cis‐diamminedichloroplatinum (II) (cis‐DDP), 6 mg/kg. A significant increase in total volume of urothelium was found in the group treated with CTX combined with radiation compared with the control. The total volume of connective tissue in the muscular layer in the groups which received X‐rays only showed a significant increase compared to the control. There also seemed to be an increase in the amount of connective tissue in muscular tissue, although this was not statistically significant, in the groups treated with CTX and irradiation compared with CTX alone. This indicates the importance of radiation in the development of late damage. Results from bladders treated with cis‐DDP alone or combined with radiation were statistically inconclusive due to the small number of animals in each group. The group treated with CTX and radiation (15 Gy) showed a significant decrease in the total volume of lamina propria, but no difference was found in the other treatment groups when comparing connective tissue, the muscular layer or the amount of vessels and capillaries with their corresponding control groups. Stereological analysis seems a feasible method to demonstrate late bladder tissue changes encountered after irradiation alone or combined with chemotherapy.
Porcine membranoproliferative glomerulonephritis with intramembranous dense deposits (porcine dense deposit disease)
Authors: JANSEN JOHAN HØGSET
Published: August 2009

A membranoproliferative glomerulonephritis (MPGN) was diagnosed in 10 Yorkshire piglets, aged between 33 and 73 days, that had been submitted for routine autopsy. Matings of animals related to these piglets subsequently produced 15 offspring that had a membranoproliferative glomerulonephritis when examined at between 5 and 48 days of age. The principal light microscopic changes in all examined glomeruli of all affected piglets were thickening of the glomerular capillary walls and a conspicuous mesangial cell proliferation. The glomerular capillary walls of the affected piglets possessed a weak but distinct autofluorescence when stained with thioflavine T and a strong reactivity for complement factor 3 as demonstrated with immunohistochemical techniques. Ultrastructural examination revealed intramembranous dense deposits in the glomerular basement membranes of kidneys from all affected piglets. On the basis of the initial matings and the morphological observations, it is concluded that the described MPGN in Yorkshire piglets represents an inherited renal disease that is comparable to human MPGN type II – dense deposit disease.
Screening for cancer of the uterine cervix in Greenland
Authors: NIELSEN NILS HØJGAARD , JENSEN HENNING
Published: August 2009

During 1986–1991 diagnostic Pap smear examinations were performed in 10,998 Greenlandic women, corresponding to 60 per cent of all women over 14 years of age. Of these, more than half had at least two smears taken. Three‐year screening coverage of women over 14 was 40 per cent. Screening intensity was highest in the 25–39 age group, where more than half the population was covered. Compared to the previous 10‐year period, rates for all precursor lesions were considerably lower, most pronounced for carcinoma in situ. Such findings probably reflect previous screening experience rather than reduced background risk. However, cytological screening activity failed to reduce the incidence of invasive cancer – one of the world's highest. Most cancer patients had slipped through the system because of failure to be screened at all (57%) or failure of adequate follow‐up (23%). Infrequent screening and possible false‐negative results seem to account for most of the remaining cases. Cancer patients with a history of failed follow‐up reflected an inadequate follow‐up management over the last 15 years of women with abnormal smear results. The findings of this study were similar to results for 1976–1985. A nationwide, centrally organized mass screening programme is recommended.
Age‐dependent antichlamydial activity of human saliva
Authors: MAHMOUD ELGAALI A. , FRÖMAN GUNNAR , GENÇ MEHMET , MÅRDH PER‐ANDERS
Published: August 2009

Saliva samples from 51 children ranging from 1 month to 12 years of age were studied for their effect on the capacity of Chlamydia trachomatis, serotypes I and L2 elementary bodies (EB), to form inclusions in cycloheximide‐treated McCoy cell cultures. The results were compared to those of tests using saliva from adults. No antibodies against C. trachomatis or Chlamydia pneumoniae could be detected by microimmunofluorescence tests in either group. Saliva of children between 1–4 years of age showed an age‐related decrease in the chlamydial inclusion count (i.e.). Saliva from children older than 4 years of age, like saliva from healthy adults, showed a pronounced reduction of the i.e. (up to 70%). The study indicates that children between 1–4 years gradually develop a natural antichlamydial activity against C. trachomatis, and above that age they exhibit the same level of antichlamydial activity as adults. The inhibitory activity was heat‐resistant.
Detecting methicillin‐resistant Staphylococcus epidermidis– disc diffusion, broth breakpoint or polymerase chain reaction?
Authors: HEDIN GÖRAN , LÖFDAHL SVEN
Published: August 2009

Growth conditions are important for the expression of resistance to methicillin among staphylococci. Consequently a phenotypic susceptibility test has to be chosen carefully to avoid false susceptible results. In this study we wanted to devise rapid and simple phenotypic tests whose results completely correlate with the presence of the methicillin resistance gene, mecA. A simplified polymerase chain reaction (PCR) method not needing separate DNA extraction from the tested bacteria was used to amplify a 449 bp region of the mecA gene. One hundred and ten strains of S. epidermidis were tested. The results were in complete agreement with those from a broth tube breakpoint test, known to identify more strains as resistant than does the method recommended by NCCLS. In disc diffusion tests it was possible to clearly distinguish resistant from susceptible strains by using discs containing oxacillin, cephalexin and cephradine. A 5 μg cephradine disc was further analysed by testing another 441 consecutive clinical isolates of staphylococci. All resistant coagulase‐negative staphylococci grew out to the edge of this disc, whereas susceptible strains showed an inhibition zone at least 10 mm in diameter. The 5 μg cephradine disc is recommended for routine work. The PCR method and broth tube breakpoint test are both reliable reference methods.
Tissue distribution of human IgG Fc receptors CD16, CD32 and CD64: An immunohistochemical study
Authors: TUIJNMAN WOUTER B. , VAN WICHEN DICK F. , SCHUURMAN HENK‐JAN
Published: August 2009

The tissue distribution of human IgG Fc receptors (FcγRs) classified in three clusters of differentiation (CD16, using 5 antibodies, CD32, using 2 antibodies; and CD64, using 3 antibodies) was evaluated by immunohistochemistry on lymphoid (lymph node, spleen, thymus, tonsil) and non‐lymphoid (heart, jejunum, kidney, liver, lung, muscle, stomach, and skin) tissues. Macrophage‐like cells, including Kupffer cells, expressed all three classes of FcγR. Part of the cells coexpressed HLA‐DR. Interdigitating dendritic cells that were present in high density in interfollicular areas of a lymph node showing dermatopathic lymphadenopathy were immunoreactive for CD32, but not for CD16 or CD64 antibodies. In lymphoid tissue, mantle zones of secondary follicles were labelled by CD32 and some CD16 antibodies. The immunolabelling of mantle zones was not present after washing the sections at low pH, which suggests that the molecules detected were passively absorbed on the cell surface (i.e. soluble FcγR). The immunolabelling of tonsil sections by various CD16 antibodies showed three patterns. The first (anti‐Leu‐11b) revealed labelling of solitary macrophage‐like cells. The second (BW209/2 and 3G8) revealed, in addition, labelling of germinal centres. The third (CLBgranI and CLBgranII) revealed labelling of solitary cells and follicle mantles. This labelling on tissue sections was also seen in the analysis of follicular dendritic cells isolated from tonsil. The cells were faintly immunoreactive for 3G8, as well as for CD16 mAb CLBgranl, and both CD32 mAbs. In all tissues investigated there was immunoreactivity for FcγRs in varying intensity on endothelial cells of blood vessels.
Immunohistochemical localization of an inducible form of nitric oxide synthase in various organs of rats treated with Propionibacterium acnes and lipopolysaccharide
Authors: BANDALETOVA TATIANA , BROUET ISABELLE , BARTSCH HELMUT , SUGIMURA TAKASHI , ESUMI HIROYASU , OHSHIMA HIROSHI
Published: August 2009

Immunohistochemical localization of an endotoxin‐inducible form of nitric oxide synthase was examined using rabbit polyclonal antibody against the enzyme purified from rat liver. In rats treated with Propionibacterium acnes and lipopolysaccharide, immunostaining was observed in macrophages, occasional lymphocytes, neutrophils and eosinophils in red pulp of spleen, Kupffer cells, endothelial cells and hepatocytes in liver, alveolar macrophages in lung, macrophages and endothelial cells in adrenal glands, and histiocytes, eosinophils, mast cells and endothelial cells in colon. Immunoreactivity was also evident in the following tissues: histiocytes and endothelial cells in kidney; histiocytes and neutrophils in esophagus; macrophages and eosinophils in duodenum; macrophages, some lymphocytes and mast cells in ileum; histiocytes in thymus; and endothelial cells in heart and aorta. Immunoreactivity was not detected in these organs from untreated rats. Positively staining cells in these rat organs appeared within 2.5 h after lipopolysaccharide administration; their number dramatically increased within the next 2.5 h, remained at high levels for a further 19 h, and then decreased over the following 24 h. The number of positive cells appearing correlated well with the nitric oxide synthase activity biochemically determined in the same organs.
Biological properties of bacterial peptidoglycan
Authors: JOHANNSEN LARS
Published: August 2009

Synthetic and natural muramyl peptides have a variety of biological actions in mammals, including the abilities to enhance sleep and body temperature. Although muramyl peptides can be detected constitutively in mammalian organisms, no biochemical synthetic pathways are known for muramyl peptides in mammals. However, muramyl peptides are well known as components of bacterial cell wall peptidoglycan (synonym: murein). Isolated bacterial cell walls elicit host responses similar to those produced by bacterial infections or by purified muramyl peptides. Mammalian cells which phagocytize bacteria can digest bacterial cell walls and release biologically active muramyl peptides. The released muramyl peptides then express some or all of the biological effects observed with synthetic muramyl peptides. Also, cell‐free systems consisting of isolated bacterial cell walls and lysozyme produce substances with similar biological activities.
Role of x‐linked immunodeficiency (xid) and NK activity in rejection of human tumor xenotransplants in nude mice
Authors: MARUO KOHJI , SHIMAMURA KAZUO , HIOKI KYOJI , ITOH MAMORU , UEYAMA YOSHITO , TAMAOKI NORIKAZU
Published: August 2009

In order to elucidate the factors influencing the takes of human tumor xenografts in nude mice, we compared the transplantability of human tumors in nude mice with additional genetic defects in the immune system. The nude mice strains tested were classified as follows by expression of the beige gene and the x‐linked immunodeficiency (xid) gene: 1) high NK nude (C57BL/6N, nu/nu), 2) low NK nude (C57BL/6 bg/bg nu/nu), 3) high NK nude with B‐cell defect (CBA/N nu/nu), and 4) low NK nude with B‐cell defect (NIH(S)III). Takes of human tumor xenografts including gastric carcinoma, T‐cell lymphoma and B‐cell lymphoma were better in nude mice with xid (CBA/N and NIH(S) III nude mice) than in nude mice without xid (B6 and beige nude mice). In addition, among the nude mice with xid expression, the takes were slightly better in nude mice with a CBA/N background than in those with a NIH(S) background. Moreover, the xenotransplantation rate in (CBA/N × C57BL/6N)F1 male nude mice with xid expression was higher than in (C57BL/6N × CBA/N)F1 males without xid expression, but did not react the same level as that in CBA/N nude. On the other hand, introduction of the beige gene into nude mice minimally improved the takes of human tumor xenografts under limited experimental conditions (inoculation of 100 times 105 T‐cell lymphoma and 1 times 105 gastric carcinoma cells) despite the reduction of NK activity. In xenotransplantation of human tumors directly from patients, the take rates of the tumors were also better in CBA/N nude mice than in BALB/cA nude mice. The results in the present report confirmed the effect of xid and CBA/N genetic background on human tumor xenografts in nude mice, suggesting the existence of serum factors, possibly present in serum IgM, mediating rejection of the xenografts.
Serum immunoglobulin (G, A and M) classes and IgG subclasses in sickle cell anaemia
Authors: HEDO CHIKE , AKEN'OVA YETUNDE , OKPALA IHEANYI , SALIMONU LEKAN
Published: August 2009

Serum concentrations of immunoglobulins G, A, M and IgG subclasses were determined by single radial immunodiffusion assay in a population of sickle cell anaemia patients resident in the tropics. Fifty apparently healthy subjects of haemoglobin genotype AA, of comparable age, sex and socioeconomic status (SES), and in the same environment as the patients, were included as controls. Three indices of morbidity in SCA, namely frequency of crisis, degree of anaemia and the number of organ complications, were used to derive a severity score for each patient; and thus categorize the subjects into severity groups. Immunoglobulin levels were then correlated with the indices of morbidity as well as the derived severity score. IgG, IgA, IgM, IgG1 and IgG3 levels were significantly raised in the SCA subjects when they were compared as a group with the controls. When separated into disease severity groups, the mildly affected patients were found to have virtually normal levels of immunoglobulins. Total IgG concentration and level of the IgG3 subclass showed significant positive correlation with frequency of crisis and derived severity score. Markedly raised levels of IgG and IgG3 may be predictive of severity in sickle cell anaemia.
Prognostic value of nucleolar size and size pleomorphism in choroidal melanomas
Authors: SØRENSEN FLEMMING BRANDT , GAMEL JOHN W. , JENSEN OVE A. , LADEKARL MORTEN , McCURDY JEFF
Published: August 2009

Morphometric estimates of nucleolar size have been shown to possess a high prognostic value in patients with uveal melanomas. The authors investigated various quantitative estimators of the mean size and pleomorphism of nucleoli in choroidal melanomas from a consecutive series of 95 Danish patients treated solely by enucleation of the melanoma‐bearing eye in the period from 1977 to 1979. Theindependentprognostic value of the quantitative histopathological variables was evaluated using multivariate Cox analysis. Apart from patient data and information about tumor extent, histological type of melanoma, and largest macroscopic tumor dimension (LTD), the following histomorphometric estimates were obtained: mean diameter of the 10 largest nucleoli (MLN), point‐sampled mean nucleolar profile area (nucleolar āo) and the associated standard deviation of nucleolar āo, the volume‐weighted mean nucleolar volume (nucleolar v̄v), and associated variance and relative variance of nucleolar volume (nucleolar Varv(v) and nucleolar CVv(v), respectively). The intra‐observer reproducibility of estimates of nucleolar v̄vwas tested in 20 cases and found acceptable (r = 0.84, 2p = 3.6 · 10−6). Correlation analysis indicated that estimates of nucleolar v̄vand nucleolar Varv(v) are identical in choroidal melanomas, except for a scaling factor. There were no differences of the grouped means of histomorphometric estimates in choroidal melanomas that showed intra‐ or extra‐ocular extension (2p>0.13). LTD was of the same magnitude in all histological types of melanoma (2p = 0.69). Estimates of nucleolar v̄vand associated estimates of nucleolar volume pleomorphism were increased in melanomas of the mixed and epitheloid cell type (2p<0.005). Single factor analyses showed that all quantitative histopathological variables were of prognostic value in the studied patients, in addition to the histological type of tumor. However, using multivariate analysis, only estimates of LTD, MLN, nucleolar v̄v, and of nucleolar volume pleomorphism had independent prognostic value.
Enzyme changes in remodelling epithelial cells: A histochemical study of the rat jejunum in vivo during and following exposure to deoxycholic acid
Authors: MATOVELO JAYRO AMOS , LANDSVERK THOR , SUND REIDAR BREDO
Published: August 2009

Loops of rat jejunum were exposed in vivo to different concentrations of deoxycholic acid (DOC; 0, 2.5, 5, 10 and 20 mM). Following a 30 min exposure period, DOC was washed out of the loops and the intestines were allowed to recover for 15 or 150 min. Frozen tissue for enzyme histochemistry was collected during exposure and following the recovery periods. As shown previously, exposure to DOC caused a dose‐dependent loss of epithelial cells at the villous tips and denudation of the lamina propria. Flattened epithelial cells bordering the denuded areas were, however, responsible for a rapid restoration of epithelial continuity, which was completed within 15 min. In the present study, these flattened cells showed normal reactivity for non‐specific esterase and succinate dehydrogenase. In contrast, following a prolonged recovery period (150 min), a subpopulation of enterocytes at the villous tips that otherwise appeared normal showed decreased reactivity for brush border enzymes and non‐specific esterase, and a positive reaction for mucin. A shutdown in the synthesis of cytoplasmic enzymes and redistribution of cell surface enzymes could be responsible for these late occuring enzyme changes, that were consistently observed after 150 min of recovery from DOC at 20 mM. Alternatively, retention of goblet cells and/or a modification in enzyme synthesis may explain the presence of mucin that was demonstrated in the epithelial cells which had low enzyme reactivity.
Classification of tubulo‐papillary renal cortical tumours using estimates of nuclear volume
Authors: BROOKS BRENDA , SØRENSEN FLEMMING BRANDT , OLSEN STEEN , HOLM‐NIELSEN PETER
Published: August 2009

The classification of renal cortical tumours is problematic, with no clear division of benign from malignant tumours. Unbiased stereological estimates of volume‐weighted nuclear volume (nuclearv̄v) were obtained by point sampling of nuclear intercepts in a retrospective study of 36 variably sized tubulo‐papillary basophilic cell renal cortical tumours. There was no clear pattern of evolution of nuclearv̄v with increasing macroscopic tumour diameter. Estimates of nuclearv̄v could not distinguish between 21 tumours classified as renal adenomas with macroscopic diameters < 3 cm (average nuclearv̄v= 241 μm3) and 15 tumours classified as renal cell carcinomas with diameters > 3 cm, or aggressive histological pattern (average nuclearv̄v= 229 μm3) (2p= 0.68). In this subtype of renal cortical tumours, estimates of nuclearv̄v do not support the historical convention of using a 3 cm tumour diameter as the dividing line between adenomas and carcinomas, but support the theory of a single group of tumours. As most of the truly incidental renal cortical tumours are less than 1 cm in diameter, this limit could be considered. Such small benign cortical nodules have never been reported to metastasize, and would thus be excluded from being diagnosed and registered as malignant. Although this dividing line is again arbitrary, and cannot be justified by the stereological measurements, it is a practical solution to a clinical problem. There were too few examples of disease progression to assess the prognostic significance of nuclearv̄v in these tumours.
Modulation of the effect of humoral‐mediated cytotoxicity on isolated rat pancreatic islets by glucose
Authors: SCHRÖDER D. , KOHNERT K. D. , HEHMKE B. , BESCH W. , SCHMIDT S.
Published: August 2009

Rat islets of Langerhans exposed for 20 h at high glucose (20 mmol/l) to 50% or 20% experimentally raised rabbit anti‐rat islet cell surface antiserum (ICSA‐positive serum) plus complement exhibited an irreversible loss of glucose‐stimulated insulin secretion. In contrast, islets treated with 50% ICSA‐positive serum at low glucose (5.5 mmol/l) could overcome this alteration within a subsequent 48 h recovery period at 10 mmol/l glucose in the absence of ICSA, and islets affected at 5.5 mmol/l glucose by 20% ICSA‐positive serum even retained the insulin secretory potential and responded on glucose challenge already immediately after the removal of ICSA. The islet insulin content was reduced by the effect of 50% as well as of 20% ICSA‐positive serum and complement irrespective of whether the glucose level amounted to 5.5 or 20 mmol/l during serum influence. However, islets altered in a normoglycaemic environment at 5.5 mmol/l glucose by 20% ICSA‐positive serum restored their insulin content up to the level of control islets, whereas those islets affected under hyperglycaemic conditions at 20 mmol/l glucose only partially recovered. Thus, β‐cell loss and/or impairment of the insulin secretory mechanisms result from the simultaneous action of humoral‐mediated anti‐islet cytotoxicity and elevated glucose level, and cause the diminished insulin secretory potential of the islets. These results support the hypothesis that decreasing the insulin secretory activity of β cells may protect them from cytotoxic immunological attacks.
Defective HLA class II expression in monocytes of type 1 diabetic patients
Authors: PARKKONEN P. , HYÖTY H. , HUUPPONEN T. , LEINIKKI P. , TUOMILEHTO‐WOLF E. , KNIP M. ,   THE CHILDHOOD DIABETES IN FINLAND STUDY GROUP
Published: August 2009

Activation of T‐helper cells is modulated by the intensity of HLA class II expression on antigen‐presenting cells. We evaluated whether any abnormalities could be found in the expression of HLA‐DR and ‐DQ molecules on monocytes in type 1 diabetic subjects. DR and DQ molecules were induced by human recombinant interferon‐gamma on cultured peripheral blood monocytes obtained from children with type 1 diabetes (N = 28), their siblings (N = 18) and unrelated healthy controls (N = 21). The response in DQ induction varied considerably between different individuals, but the average responsiveness was significantly lower in patients compared to siblings and unrelated controls. In addition to the diabetic subjects deficient DQ induction was also observed in three siblings. One of them had high levels of islet cell antibodies and presented with diabetes 6 months later, and another had active rheumatoid arthritis. The response in DR induction was also slightly lower in patients than in siblings, but did not differ from that in unrelated controls. The results suggest abnormalities in the regulation of HLA class II expression in type 1 diabetic subjects possibly reflecting the ongoing autoimmune process.
Adherence of group B streptococci to human endothelial cells in vitro
Authors: KÄLLMAN JAN , SCHOLLIN JENS , HÅKANSSON STELLAN , ANDERSSON ANGELA , KIHLSTRÖM ERIK
Published: August 2009

A model using human umbilical vein endothelial cells was developed to study the adhesion of group B streptococci. Ten clinical isolates of serotypes Ia, Ib and III, including six blood isolates from neonates with early‐onset disease, were studied. Isogenic variants with different ability to express capsule substance were also included. Clinical isolates of serotype III adhered significantly better than other serotypes. Strains with high ability to adhere to endothelial cells also had high ability to bind to plastic. Isogenic variants of serotype III with low amounts of capsule substance adhered significantly better to cells than variants expressing high amounts of capsule substance. These results show that the higher adherence of serotype III strains may be a virulence factor and contribute to the finding that this serotype dominates in invasive group B streptococcal disease.
Correlations of peritoneal interleukin‐6, serum beta‐2 microglobulin and urinary beta‐2 microglobulin after elective abdominal surgery
Authors: TSUKADA KATSUHIKO , KATOH HIROYUKI , SUZUKI TANJI , TAKENOSHITA SEIICHI , NAGAMACHI YUKIO
Published: August 2009

Postoperative peritoneal interleukin‐6 (P‐IL‐6), serum beta‐2 microglobulin (SBMG) and urinary beta‐2 microglobulin (UBMG) concentrations were examined in 33 patients who underwent various types of abdominal surgery. Similar time‐dependent changes in SBMG and UBMG were observed in five other patients who underwent abdominal surgery. SBMG decreased the first day after the operation (1POD), then increased and reached the preoperative level by the third day after the operation (3POD). UBMG started to increase 1POD, reached the maximum 3POD, then decreased. The 1POD SBMG concentration was significantly correlated with P‐IL‐6, and 3POD UBMG was also significantly correlated with P‐IL‐6. These findings indicate that serum and urinary beta‐2 microglobulin concentrations are closely correlated with P‐IL‐6.
Pathways of complement (C3) activation in rabbits infected with Trypanosoma evansi
Authors: UCHE U. E. , JONES T. W.
Published: August 2009

Immunoelectrophoresis was used to evaluate the pathways of complement (C3) activation with sera from rabbits infected with Trypanosoma evansi. C3 activation via the classical pathway was of a higher magnitude than via the alternative pathway using both the intact parasite and soluble parasite antigens. This observation indicates that although the two pathways may be activated during infection, the activation of the classical pathway may be dominant in a host's response to a T. evansi infection during both the early and late stages of the disease.
Role of protein kinase C during interferon‐γ‐ and phorbol ester‐stimulated immunocytochemical expression of ICAM‐1 in human renal carcinoma cells
Authors: HANSEN ALASTAIR B. , BOUCHELOUCHE PIERRE N. , GIESE BIRGITTE N. , ANDERSEN CLAUS B.
Published: August 2009

Incubation of the human renal carcinoma cell line CaKi‐1 with interferon‐γ (IFN‐γ) or the phorbol ester, phorbol‐12‐myristate 13‐acetate (PMA) strongly stimulated the immunocytochemical expression of the intercellular adhesion molecule‐1 (ICAM‐1) in a dose‐dependent manner. Since PMA is capable of activating the Ca2+/phospholipid‐dependent protein kinase C (PKC), we investigated the role of this kinase during IFN‐γ signal transduction. Calcium ionophore A23187 significantly enhanced IFN‐γ‐ and PMA‐induced ICAM‐1 staining. While staurosporine, H7 and sphingosine, three known PKC inhibitors, blocked the PMA effect, only staurosporine abrogated the action of IFN‐γ. Finally, 24 h of PMA pretreatment with subsequent IFN‐γ stimulation enhanced ICAM‐1 staining above values from cultures where IFN‐γ was omitted. This occurred despite the fact that 24 h of PMA pretreatment abolished the effect of IFN‐γ on PKC activation, as determined by acetylated myelin basic protein 4–14 phosphorylation. In conclusion, these results suggest that additional events other than PKC activation are required for complete regulation of ICAM‐1 antigen by IFN‐γ in the whole cell population. Hence, other Ca2+‐dependent signalling pathway(s) mediated by IFN‐γ receptors must act. Further studies are needed to elucidate these specific pathway(s) activated during IFN‐γ stimulation in our model.
Antimycobacterial activity of methdilazine (Md), an antimicrobic phenothiazine
Authors: CHAKRABARTY A. N. , BHATTACHARYA C. P. , DASTIDAR SUJATA G.
Published: August 2009

Methdilazine (Md) could inhibit various Mycobacterium spp. at 5–15 μg/ml concentrations in vitro as well as in vivo. When Md was tested in combination with streptomycin (Sm), rifampicin (Rf) or methyl‐DOPA (m‐D), it showed synergistic effects only with respect to methyl‐DOPA.
Single lung transplantation Morphological surveillance by transbronchial biopsy
Authors: FOERSTER ARNOLD , BJØRTUFT ØYSTEIN , GEIRAN ODD , ROLLAG HALVOR , LEIVESTAD TORBJØRN , FRØYSAKER TOR
Published: August 2009

Seven cases of single lung transplantation are reported. The recipients were all below 60 years of age and severely disabled with end‐stage lung disease. Transplantation was performed according to ABO blood group compatibility and negative lymphocytotoxic cross‐match between donor and recipient irrespective of HLA mismatch. Recipients' diagnoses were sarcoidosis (3), alfa‐1 antitrypsin deficiency (3), and idiopathic emphysema (1). Mean recipient age was 48 ± 2.4 years (range 45–52). Donor age was 29.7 ± 5.6 years (range 16–49). The immunosuppressive regimen included cyclosporin A, azathioprine, steroids and rabbit antithymocyte globulin. Excellent graft function was achieved. Six patients survived the postoperative period and are alive 4–18 months posttransplant. One patient died after the operation due to pneumonia with respiratory distress syndrome. Graft function was also monitored by transbronchial biopsy, and 57 biopsy procedures were performed without fatal complications. Acute cellular rejection was seen in 16 biopsy specimens from 5 recipients (grade 1 and 2 rejection in 14, grade 3 rejection in 2). Neither severe rejection with septal necrosis (grade 4) nor obliterative bronchiolitis was seen. The rejection rate was 0.03 episodes per patient/month. In contrast to other reports, episodes of cellular rejection occurred throughout the observation period, and were not mainly limited to the first 4 months posttransplant. Graft vascular occlusive disease or chronic vascular rejection was found in 6 biopsy specimens from one recipient. Five patients experienced 7 episodes of cytomegalovirus infection. The cytomegalovirus infection rate was 0.01 episodes per patient/month. The incidence of infection was significantly lower compared to previous studies of rejection in other lung graft combinations. Both infections and rejection episodes may contribute to the development of obliterative bronchiolitis. Almost one third of the specimens (30%) showed lymphocytic bronchitis without perivascular inflammation. The absence of perivascular infiltrates and exclusion of infectious agents leaves in question the aetiology of this inflammation. The lymphocytic bronchitis could be ischaemic, related to aspiration, or represent recurrent sarcoidosis, or, in fact, express bronchial rejection. All biopsy specimens regarded as rejection with cellular infiltrates in the lung parenchyma also showed a lymphocytic bronchitis. The impact of HLA mismatch on cellular and vascular rejection is unclear. Transbronchial biopsy is a reasonably safe and reliable method in the diagnosis of rejection and infection in single lung transplantation.
Defective T‐cell stimulatory pathways in patients after allogeneic bone marrow transplantation (BMT) in man
Authors: MØLLER JARL , HOFMANN BO , JACOBSEN NIELS , RYDER LARS P. , PLATZ PER , DICKMEISS EBBE , SVEJGAARD ARNE
Published: August 2009

Immunological reconstitution after allogeneic bone marrow transplantation in man is characterized by a decreased lymphocyte transformation response to various mitogens and antigens during a period of from months to years. One reason for the decreased proliferative capability could be an inverted CD4/CD8 ratio; however, the present investigation demonstrates that this is not the only explanation for the immunodeficiency, since the CD4 as well as the CD8 subset, when studied in isolation, have qualitative defects, as evidenced by a reduced response of both subsets to stimulation with PHA, anti‐CD2 and anti‐CD3 MABs. The reason for the qualitative defect is unknown but a distorted composition of the CD4+ as well as the CD8+ T‐cell subsets is suggested by the present investigations. We also observed that the PHA response was almost completely reconstituted one year after BMT, while the PWM response was still severely affected. The present study suggests that T‐cell subsets which differ in their capacity to respond to PHA and PWM have different kinetics of reconstitution after BMT.