bluuebunny/crossref_metadata_2025 huggingface dataset:
Tumours in Iceland. 14. Malignant tumours of soft tissues
Authors: AGNARSSON BJARNI A. , BALDURSSON GUDJÓN , BENEDIKTSDÓTTIR KRISTRÚN R. , HRAFNKELSSON JÓN
Published: August 2009

All malignant tumours of soft tissues diagnosed in Iceland between 1955 and 1988 were reviewed histologically. Of a total of 155 tumours, 129 were, on review, soft tissue sarcomas, while 26 had been erroneously diagnosed and were excluded from the study. In 25% of the tumours the original sarcoma diagnosis was changed to another sarcoma type. Malignant fibrous histiocytoma, liposarcoma and leiomyosarcoma together comprise more than half of all tumours diagnosed. Contrary to previous estimations, according to this study, the age‐standardized incidence of soft tissue sarcoma in Iceland is 1.8 per 1000.000 for males and 1.6 per 100.000 per females, which is similar to incidence rates in the other Nordic countries. The majority of sarcomas were of relatively high grade. The tumours were graded using both three and four grades of malignancy. Both systems yielded prognostic information although it was not possible to detect significant differences in survival for grade I and grade II tumours when the four grade system was used. The results of this study show that the epidemiology of soft tissue sarcoma in Iceland is similar to that found in other Western countries.
Langerhans cells and subsets of lymphocytes in the nasal mucosa
Authors: HELLQUIST H. B. , OLSEN K. E. , IRANDER K. , KARLSSON E. , ÖDKVIST L. M.
Published: August 2009

Langerhans cells and different lymphocytes were studied in the nasal mucosa of 39 woodwork teachers and a control group of 14 healthy subjects. Ten of the woodwork teachers were sensitized as determined by skin prick test. A panel of different monoclonal antibodies was applied on the frozen nasal mucosal specimens. Intraepithelial CD1‐positive dendritic cells were found in all specimens. However, there was no difference between the number of these Langerhans cells found in the study group and the number found in the controls. In every specimen the intraepithelial lymphocyte population was dominated by T lymphocytes, and there were relatively few B cells. Similarly the ratio between CD4‐and CD8‐positive lymphocytes in the study group and the controls was the same. In all specimens there was a dominance of T suppressor/cytotoxic cells compared with T helper/inducer cells. The study confirms that Langerhans cells are present in normal nasal surface epithelium, and suggests that there is no basic difference in the number of Langerhans cells between healthy persons, persons with nasal complaints, and persons with nasal allergy. The dominance of T lymphocytes in the epithelium may indicate the existence of a local cell‐mediated immunity other than that associated with the regulation of IgE.
Detection of Chlamydia trachomatis in first‐void urine collected from men and women attending a venereal clinic
Authors: GENÇ MEHMET , STARY ANGELICA , BERGMAN SUZANNE , MÅDH PER‐ANDERS
Published: August 2009

Cervical, urethral and first‐void urine (FVU) specimens from 196 men and 245 women attending a venereal outpatient clinic were studied by culture and a commercial enzyme immunoassay (EIA) (CHLAMYDIAZYME®). Confirmatory chlamydial testing by a direct fluorescence assay (DFA) (MICROTRAK®) was performed on the sediments of the positive EIA samples from culture‐negative patients. Chlamydia trachomatis was isolated from 11% of the men and 12% of the women. Of the women, 67% were positive in both sampling sites and 33% in the cervix only. No further cases were found when a female urethral swab was cultured. All the chlamydia‐positive urine samples were obtained from women who were positive in the urethra. The denominator used to calculate sensitivities was the combination of patients with culture‐ and EIA‐positive results which could be confirmed by DFA. The sensitivity of our culture method was 85% for men and 77% for women. In men, the sensitivity of EIA was greater on urine than on urethral specimens (77% vs 62%; p<0.1). In women, the sensitivity of EIA on urine was significantly poorer than that on cervical specimens (54% vs 85%, p<0.001). The specificity of EIA ranged between 94 and 100%. Our study suggests that it may be worth using FVU in a trial for the diagnosis of genital chlamydial infections in symptomatic men, but not in symptomatic women.
Post‐streptococcal glomerulonephritis: studies on the interaction betwen nephritis strain‐associated protein (NSAP), complement and the glomerulus
Authors: PEAKE PHILIP W. , PUSSELL BRUCE A. , KARPLUS THOMAS E. , RILEY ELIZABETH H. , CHARLESWORTH JOHN A.
Published: August 2009

Nephritis strain‐associated protein (NSAP), a streptokinase produced by strains of streptococci isolated from patients with acute glomerulonephritis, is believed to be a specific antigen which participates in the production of glomerular injury. In order to investigate the mechanisms by which NSAP induces damage we have examined its potential to activate complement in vitro and to bind to isolated human glomeruli. NSAP, both alone and in combination with specific antibody, caused depletion of complement in normal human serum as measured by total haemolytic complement activity and generation of the complement breakdown products, C3a and C4a. Furthermore, Scatchard analysis showed that NSAP bound tightly to human glomeruli (Ka of 400±240 times 106 M) when compared to non‐nephritic streptokinase (Ka of 7.3±4.1 times 106 M) and fully cationized human serum albumin (Ka of 0.6±0.04 times 106 M). These findings are consistent with the hypothesis that the deposition of streptococcal antigens within the glomerulus may precede the fixation of complement and specific antibody.
ELISA for detection of Mycoplasma pneumoniae antigens using monoclonal antibodies
Authors: HIRSCHBERG LOTTA , HOLME TORD
Published: August 2009

Seven monoclonal antibodies directed against major antigens of Mycoplasma pneumoniae were selected for the development of an antigen detection assay. Three of these were directed to the 170.000‐dalton adhesin of M. pneumoniae. The test was an antigen‐capture enzyme immunoassay using the different monoclonal antibodies for capture of antigen and a polyclonal rabbit antiserum as detection reagent. With three of the monoclonal antibodies a detection limit of approximately 2 ng M. pneumoniae protein was obtained, as determined by titration of M. pneumoniae organisms in buffer. The detection limit of the assays was only slightly less when the other four monoclonal antibodies were used. In artificially infected nasopharyngeal aspirates the detection limit was approximately 10 times lower. The fact that no significant differences in the detection limit of the assays were recorded using monoclonal antibodies directed against different antigens indicates that these antigens were available for reaction with antibodies irrespective of their location in intact M. pneumoniae cells. In the assay there were no significant cross‐reactions with a number of bacterial species potentially colonizing the respiratory tract, except for a protein A‐positive strain of Staphylococcus aureus. Our test is equally sensitive to another recently described ELISA using polyclonal antibodies. In comparison with other recommended methods such as immunoblot and culture‐amplified antigen detection assays, the ELISA is more rapid and less laborious.
How do animal phagocytes process bacterial lipopolysaccharides?
Authors: MUNFORD ROBERT S.
Published: August 2009

Phagocytic cells, the front‐line defense against bacterial invasion, have enzymes that deacylate and dephosphorylate the toxic lipid A moiety of gram‐negative bacterial lipopolysaccharide (LPS, also called endotoxin). These enzymes may detoxify LPS without destroying its immunogenicity or antigenicity ‐ in fact, the polysaccharide region of LPS often resists degradation in vivo and retains its antigenic epitopes for long periods of time. The net result, detoxification of LPS with preservation of its immunogenic and antigenic properties, may prevent excessive inflammatory responses while promoting the development of antibacterial immunity.
Properties of a deep Proteus R mutant isolated from clinical material
Authors: KRAJEWSKA‐PIETRASIK DANUTA , RÓZALSKI ANTONI , BARTODZIEJSKA BEATA , RADZIEJEWSKA‐LEBRECHT JOANNA , MAYER HUBERT , KOTELKO KRYSTYNA
Published: August 2009

Some biological features of a deep P. mirabilis 17301 R mutant isolated from the urine of a patient with chronic UTI were studied and compared with similar features of P. mirabilis S forms and five induced Proteus R mutants of different chemotypes. There were no differences in lethal toxicity and adhesion to human uroepithelial cells. Of all the R mutants tested, two of them, 17301 and R4, exhibited strong cell‐bound hemolytic activity. The P. mirabilis R 17301 was characterized as the most invasive (tested in L929 mouse fibroblasts) compared to the other Proteus S and R forms. The structure of PS from a clinical R mutant investigated and the results of serological studies prove that this mutant belongs to the Rc chemotype.
Genomic groups and biochemical profiles of clinical isolates of Enterobacter cloacae
Authors: LINDH E. , URSING J.
Published: August 2009

A collection of 123 clinical strains presumptively identified as Enterobacter cloacae and 12 type and reference strains of Enterobacter spp. were genotypically investigated by a quantitative bacterial dot method for DNA‐DNA hybridization, giving an estimate of Δm (difference in thermal denaturation midpoint between homologous and heterologous duplexes). The API 20E system was used for phenotypic characterization. Using discontinuities in the values of Δm as criterion, five genomic groups of E. cloacae could be demonstrated, eleven ungrouped strains representing at least one additional group. Nine API profiles were found, the ideal phenotype of E. cloacae (i.e. the phenotype showing the most common reaction for the species in all tests studied) being the most frequently found. The type strain of E. cloacae and the reference strain CDC 1347–71 represented rather small genomic groups of three and eight strains respectively, most of them inositol positive. A majority of 98 isolates formed one single genomic group, biochemically dominated by the ideal phenotype. The genomic groups could not be differentiated phenotypically. At present there seems to be no reason for an attempt to split E. cloacae into two or more species. The type strain of E. dissolvens showed itself to be closely related to the type strain of E. cloacae (Δm 2.3°C), indicating that the two species may be regarded as subjective synonyms.
Human antibody response to the major adhesin of Mycoplasma pneumoniae: Increase in titers against synthetic peptides in patients with pneumonia
Authors: HIRSCHBERG L. , HOLME T. , KROOK A.
Published: August 2009

Peptides corresponding to parts of the P1 protein (major adhesin) of Mycoplasma pneumoniae were synthesized. On the basis of predicted antigenicity, seven sequences containing 17 to 21 amino acids were selected. In addition, one peptide containing a sequence of 13 amino acids shown to be related to cytadherence of M. pneumoniae was included. Serum samples from 56 patients with pneumonia were tested for a rise in titers of specific IgG during infection, using the peptides as coating antigens in ELISA. A titer rise against one or more peptides was observed in 10 out of 13 patients with serological evidence of mycoplasmal etiology. Specific antibodies to two or more peptides were demonstrated in three patients, whereas seven patients responded to one peptide only. In the sera from patients with pncumonia of non‐mycoplasmal etiology, no titer rises above the cut‐off level were observed. Our results indicate that a combination of four peptides would be possible for use as antigen for serological diagnosis of infections with M. pneumoniae.
The effect of antistaphylococcal agents used alone and in combinations on the survival of Staphylococcus aureus ingested by human polymorphonuclear leukocytes
Authors: RØDER BENT L. , FORSGREN ARNE , GUTSCHIK ERNÖ
Published: August 2009

The intracellular activity of a number of drugs used alone and in combinations against Staphylococcus aureus was investigated using an experimental design which imitates the clinical situation and differs from other published methods. Staphylococci were phagocytosed by human polymorphonuclear leukocytes and, after differential centrifugation and washing, the granulocytes were incubated in 90% pooled human serum with clinically relevant drug concentrations. When exposed to antibiotics, more than 40 50% of the bacteria were located intracellularly. Fusidic acid (100 mg/1), erythromycin (20 mg/1), and clindamycin (20 mg/1) all had a bacteriostatic effect during the first 6 h of incubation, whereas rifampicin (1 and 5 mg/1), vancomycin (5 and 20 mg/1), and ciprofloxacin (2 mg/1) all acted bactericidally with decreases in viable counts between 1.3–1.9 log10. The greatest bactericidal effect was achieved with tobramycin (10 mg/1), which produced more than a 4 log10 decrease in viable counts at 6 h. Combinations of fusidic acid with other antibiotics all resulted in killing kinetics different from those achieved with the drugs used individually. The bactericidal effect of ciprofloxacin and dicloxacillin during the first 6 h was abolished when these drugs were combined with fusidic acid. However, at 24 h no significant difference was found between the effect of dicloxacillin alone versus the combination dicloxacillin and fusidic acid. The combination of fusidic acid and rifampicin resulted in a killing identical to that achieved with rifampicin used alone during the first 6 h, but at 24 h the killing by the combination was significantly greater. The bactericidal effect of the combination dicloxacillin (20 mg/1) and tobramycin (10 mg/1) equalled that obtained with tobramycin (10 mg/1) used alone. Rifampicin (5 mg/1) antagonized the bactericidal effect of ciprofloxacin (2 mg/1) during the first 6 h of incubation but at 24 h the combination acted synergistically. The results obtained are partly in agreement and partly in conflict with previous results.
Characterization of β‐lactam‐resistant Klebsiella oxytoca isolated in a neonatal intensive care unit
Authors: JALAKAS‐PÖRNULL K. , DORNBUSCH K. , KÜHN I. , RANSJÖ U. , JONSSON C. , BROBERGER U.
Published: August 2009

The occurrence of Klebsiella oxytoca resistant to ampicillin, piperacillin, aztreonam and cefuroxime in a neonatal intensive care unit, including two cases of septicemia, was shown to consist of a spread on three consecutive occasions caused by three different biochemical Klebsiella oxytoca phenotypes. All isolates, except six surface isolates from one infant belonging to phenotype 1, were sensitive to cefotaxime (MIC 0.5–4 mg/1) and ceftazidime (MIC 0.25–1 mg/1). Isolates of phenotypes 1 and 2 produced a β‐lactamase with an isoelectric point of 5.5 and isolates of phenotype 3, a β‐lactamase with an isoelectric point of 7.9. The β‐lactamases of all three phenotypes hydrolysed benzylpenicillin and more slowly cephalothin. All phenotype 1 isolates carried a 2.9 Md plasmid and most isolates also a 36 Md plasmid. All phenotype 2 isolates carried a 4.8 Md plasmid and one isolate also a 30 Md plasmid. The phenotype 3 isolates carried only one 85 Md plasmid.
Bacteriological autopsy II. Comparison of a group of cancer patients with a control group
Authors: LARSEN KNUD ERIK , SØGAARD PER , BUHL LINE , LOU HANNE ELISABETH , HENRIQUES ULRIK
Published: August 2009

A study of the results of bacteriological autopsy in a group of cancer patients compared with a control group was carried out. Autopsy was performed on 42 bodies within 24 hours of death. Thirty‐two patients had died with a cancer and the remaining ten had died of cardiovascular disease. Tissue for microscopy and material and culture were taken from the same sites. The results of the inoculations were semiquantified, resulting in a culture score, and the results of the microscopy were classified according to presence of bacteria and kind of inflammation. Furthermore, grading was done according to the degree of inflammation and whether bacteria were seen at microscopy. The culture scores were significantly higher in the cancer group than in the control group. We found a significant correlation between score and presence of acute inflammation. Likewise, a significant correlation was found between score and presence of bacteria at microscopy. As a difference between the cancer group and the control group was seen, further research on bacteriological autopsy seems worthwhile.
An antigenic determinant is shared by psoriasis‐associated p27 antigen and the Fc part of human IgG
Authors: ÅSBAKK KJETIL , RØDAHL EYVIND , LYSVAND HILDE , IVERSEN OLE‐JAN
Published: August 2009

Rabbit antisera against the major internal protein, p27, of retrovirus‐like particles from psoriatic urine, and against the serologically cross‐reacting antigen, pso p27, from psoriatic scale, reacted with the Fc part of human IgG. Evidence indicating that the p27 antigen and the pso p27 antigen are identical has been presented in previous reports. A commercial antiserum against human IgG recognized a component in the pso p27‐containing solution used as the source of antigen for immunization of the rabbits. By means of monoclonal antibodies against the pso p27 antigen, it was demonstrated that the Fc‐reacting antibodies, and the antiserum against human IgG, recognized an epitope on the pso p27 antigen. The data indicated that an antigenic determinant is shared by the p27 antigen(s) and human IgG, suggesting that p27 antigen(s) may act as antigen(s) eliciting the production of antibodies with rheumatoid factor activity in psoriatic patients.
Cyclosporine‐induced renal morphologic and immunohistologic changes in patients with chronic uveitis
Authors: ANDERSEN C. B. , LARSEN S. , ELMGREEN J. , TVEDE N. , ANDERSEN V.
Published: August 2009

Ten long‐term cyclosporine‐treated patients with chronic uveitis underwent percutaneous renal biopsy in order to evaluate a) abnormalities of renal morphology and b) the nature of lymphocytic infiltrates by immunohistochemistry. Pretransplant renal biopsies from eleven cadaveric donors served as controls. Eight of the ten patients had lymphocytic infiltrates consisting predominantly of T lymphocytes with a CD4+/CD8+ ratio of 1.46, which is identical to that of peripheral blood in healthy donors. Evidence of immune activation as estimated by the presence of interleukin‐2 receptors, transferrin receptors or by the expression of MHC class 11 antigens was not demonstrated in any of the patients. The severity of morphologic alterations did not correlate with any of the clinical or paraclinical data. Percutaneous renal biopsy should be performed within 18 months of treatment to identify patients susceptible to renal side effects of cyclosporine.
Natural killer cell function and malignant cell phenotype in hairy cell leukaemia
Authors: ÖGMUNDSDÓTTIR H. M. , THORSTEINSSON L. , SIGFÚSSON A. , SVEINSDOTTIR S. , BJÖRNSSON S. , EYJÓLFSSON G. , JÓHANNESSON G. M. , JENSSON Ó.
Published: August 2009

We have followed for 33 months the changes that occurred in natural killer (NK) cell numbers and activity in a patient (A) with hairy cell leukaemia (HCL), using a single cell assay and a microcytotoxicity assay. The composition of the peripheral blood mononuclear cell population and malignant cell phenotype were also analysed. During this period he received treatment with interferon and his grossly enlarged spleen was removed. Four further patients were also studied, two were splenectomized and all had received treatment with interferon. In four of the five patients studied there was an apparent link between low NK activity and presence of a tumour‐infiltrated spleen, and in the fifth patient, who was aleukemic and had no splenomegaly, NK function was related to disease activity. There was no correlation between NK activity and the number of target binding (TB) cells in these five patients. IFN had little direct effect on overall NK activity, but the proportion of killing cells among TB cells was increased. Three patients showed binding of several cells to a single target. Further analysis revealed that in the patients most of the TB cells were not CD56‐positive NK cells, in contrast to TB cells from normal subjects. In patient A a large proportion (84%) of TB cells were identified as malignant cells and in patient E 15% of TB cells were malignant cells. The phenotype of the malignant cells was: CD19 +, HLA‐DR + and CD25(Tac) +, except for patient A. In this patient the hairy cells were positive for the NK marker CD56 as well as the monocyte marker CD14. Furthermore, a change occurred in phenotype as only later samples carried CD25. It is concluded that the level of NK function correlates closely with disease activity in HCL and that competitive target cell binding by malignant cells may be one cause of depressed NK‐cell function in hairy cell leukaemia.
Three‐dimensional reconstruction of human juxtaglomerular apparatus (JGA)
Authors: FARAJ A. H. , LINDOP G. B. M. , MORLEY A. R. , COLEMAN S.
Published: August 2009

A three‐dimensional and morphological study of the human JGA was undertaken to establish a background for understanding the changes in this vital apparatus during various physiological and pathological conditions. Three‐dimensional reconstruction was carried out using a computer program “GLOM”. Serial sections of normal human kidneys were used after staining with specific human renin antiserum. Three‐dimensional reconstruction revealed renin‐positive cells in the afferent and efferent arterioles and interlobular arteries away from the JGA area. A close contact was demonstrated between renin‐positive cells and the macula densa. The frequency of positively stained JGAs was significantly higher in the superficial glomeruli compared to the deep glomeruli. The high renin content of the superficial glomeruli suggests higher generation of angiotensin, which may contribute to the regulation of the GFR as proposed by other workers. This preliminary study on normal human JGA is to be extended to hypertensive and renal failure patients.
Foetal antigen 2 (FA2) in the stromal reaction induced by breast carcinoma
Authors: RASMUSSEN H. BOJE , TEISNER B. , ANDERSEN J. A. , YDE‐ANDERSEN E. , SKJØDT K. , SCHRØDER H. D.
Published: August 2009

An indirect immunoperoxidase technique was used to examine the distribution of foetal antigen 2 (FA2), a recently described basement membrane (BM)‐associated antigen, in invasive breast carcinoma (n = 34), fibroadenoma (n = 5) and normal breast tissue (n = 5), and to compare its distribution with that of laminin and collagen type IV. In normal breast tissue, FA2 was detected in the intralobular stroma as a broad band around acini and ducts, but was not present in the interlobular stroma. In areas of carcinoma in situ, FA2 was present diffusely around and in close contact with the glandular elements, the staining being more intense than that found around normal glandular structures. Two distinct patterns of FA2 distribution were found in adenocarcinomas of the breast. In the fibroblast reaction type, fibroblast staining dominated, whilst in the stromal reaction type, intense and extensive staining of the surrounding stroma dominated. Significant correlation was found between the degree of fibroblast activity and the degree of anaplasia (p=0.005). FA2 extracted from breast carcinoma tissue was shown to be immunologically identical to FA2 fractions extracted from second trimester amniotic fluid (AF). The Mr of FA2 isolated from AF was estimated to be 26 kD, whereas the Mr of FA2 extracted from breast carcinoma tissue was slightly higher. The apparent Mr under reducing conditions were higher and three bands ranging from 26 to 29 kD were seen. FA2 was found to be immunologically distinct from collagen types I, III and IV, laminin, fibronectin and fibrinogen. The increased production and widespread distribution of FA2 in breast carcinomas suggest that FA2 is involved in the stromal changes which occur in response to tumour growth and/or invasion.
Regulation of aspartate carbamoyltransferase in Neisseria and Branhamella species
Authors: JYSSUM SIDSEL
Published: August 2009

The regulatory characteristics of aspartate carbamoyltransferase (ACTase EC 2.1.3.2) from various species of Neisseria and Branhamella have been compared. Great differences in the regulatory nature of the enzymes were observed. ATP and GTP were positive effectors in Neisseria meningitidis, N. gonorrhoeae and nine other coccal “true neisseriae” species. In four “false neisseriae” species, including Branhamella catarrhalis, no stimulating effect of ATP or GTP was observed. The rod‐shaped N. elongata behaved as the “false neisseriae” in these respects, despite its taxonomic affinity to the coccal “true neisseriae” species. Except in N. meningitidis and N. lactamica, CTP had no distinct stimulatory effect. CTP had a strong inhibitory effect on ACTases from N. elongata and the “false neisseriae” species N. caviae and B. catarrhalis., The inhibitory effect of CTP was weak in N. cinerea, N. denitrificans, and the “false neisseriae” species N. ovis and N. cuniculi. Thus, there was no sharp reflection of taxonomy in the regulation of ACTase by CTP in these groups of bacteria. The apparent [S]0.5 values for aspartate and carbamoyl phosphate, displayed for five of the eighteen species, showed great variability with [S]0.5 values for aspartate ranging from 6 to 34, and for carbamoyl phosphate from 2 to 9. Treatment of the enzyme from the main test microbe N. meningitidis strain Ml by heat or para‐chloromercuribenzoate (pCMB) showed that both the catalytic and the regulatory functions decreased in parallel as in the class A enzymes found in species of Pseudomonas. An estimation of the molecular weight (Mr) of the ACTase enzyme from N. meningitidis showed it to be about 295,000, which resembles the class B enzymes found in the Enterobacteriaceae.
Characterization of the a antigen of the c proteins of group B streptococci (GBS) using a murine monoclonal antibody
Authors: BEVANGER LARS , IVERSEN OLE‐JAN , NAESS AUGUSTA IRENE
Published: August 2009

A murine monoclonal antibody raised against the a antigen of the group B streptococcal c proteins was analysed by immunofluorescence and whole‐cell ELISA against a collection of 22 c protein‐producing GBS. All the strains showing fluorescence and reactivity in ELISA turned out to be a antigen‐carrying strains as defined by polyclonal rabbit antisera, while none of the strains producing only the p antigen was positive. Western blot analyses of the a antigen released into the culture medium of growing bacteria suggest that the a antigen is present as distinct proteins of variable molecular weights. The upper limit of the molecular weights varies considerably from one strain to another, from approximately 200 kD to 70 kD. With all strains, the bands seen by the MAb occurred at regularly spaced intervals of about 10 kD throughout the gel. Some strains gave rise to 15–16 bands, while others gave rise to only one or two bands. The present investigation suggests that α antigens include several, probably identical, repeating subunits of approximately 10 kD. The epitope recognized by the MAb seems to be located on a 10‐kD fragment, and in addition, it appears to be surface located, making the MAb a suitable tool in serodiagnostic work.
Monoclonal antibody 43‐9F: An immunohistochemical marker of embryonal carcinoma of the testis
Authors: VISFELDT J. , GIWERCMAN A. , SKAKKEBÆK N. E.
Published: August 2009

Discrimination between different types of germ cell tumours may be difficult in routine histological preparations. Additionally, none of the established immunohistochemical markers is completely reliable in diagnosis of embryonal carcinoma. A pilot study indicated that monoclonal antibody 43‐ 9F ‐ a marker of carcinoma‐in‐situ germ cells ‐ may also react with embryonal carcinoma of the testis. In order to elucidate the applicability of 43‐9F in diagnosis of embryonal carcinoma, 42 consecutive testicular germ cell tumours were tested immunohistochemically. Among the 42 tumours, 23 were seminomas and 19 were non‐seminomas with seminomatous components in seven of them. Embryonal carcinomas were found in 15 tumours, two being of pure type and the remaining 13 a part of mixed tumours. Additionally, the material included 11 teratomas, nine yolk sac tumours and one choriocarcinoma. Immunohistochemical stainings were performed with 43‐9F and additionally with antibodies against placental‐like alkaline phosphatase, cytokeratins, alpha‐foetoprotein and human chorionic gonadotropin. Using 43‐9F a strong colour reaction was found in 13 of the embryonal carcinomas, whereas the reaction was moderate in the remaining two cases. A weak positive reaction was found in six seminomas and the remaining 24 did not react at all. 43‐9F exhibited a positive reaction in four of 11 teratomas. The reactivity was generally weak with some focal areas with strong staining. In five cases the yolk sac tumour elements did not stain with this monoclonal antibody. The reaction was weak in three cases and in only one case was the staining intensity scored as moderate. Finally, no reaction was found in the choriocarcinoma element. Compared to the other antibodies tested, including the antibody against cytokeratins, in embryonal carcinoma immunohistochemical staining with 43‐9F was more specific, stronger and more constantly expressed. The monoclonal antibody 43‐9F may be of value in histological diagnosis of germ cell tumours. Additionally, the study confirmed the pathogenetical link between pre‐invasive carcinoma in situ and embryonal carcinoma.
Murine thymocytes with ability to inhibit I1‐2 production: I. Genetic differences between mouse strains and characterization of the model system
Authors: GIGLIOTTI DULCEAYDEE , NIHLMARK EVA LOTTA , WIGZELL HANS , HANSSON MONA
Published: August 2009

An experimental system has been established to understand the poor interleukin‐2 (I1‐2) production by activated thymocytes. This model system is further characterized here and studies were done on the possible mechanism(s) involved. Thymocytes activated by Concanavalin‐A (Con‐A), or through the CD3 complex of the T‐cell receptor (TCR), inhibit 95–99% of the I1‐2 production by spleen cells, while thymocytes stimulated by rI1‐2 or lipopolyssacharides (LPS) do not. The mechanism of inhibition is not due to production of soluble factors, consumption of available interleukin‐1 (I1‐1) or I1‐2, but is dependent on cell‐to‐cell contact. Although cellular contact is needed, cytotoxicity is not involved. Prostaglandin production is not required for the generation or exertion of the inhibitory activity. Protein and DNA synthesis are necessary for exertion of the suppressive effect. We also demonstrate a genetic difference between different mouse strains in the ability to generate the inhibitory thymocytes. Activated Balb/c thymocytes inhibit spleen cells' I1‐2 production in a non‐MHC‐restricted manner. Our studies demonstrate a regulatory capacity of activated thymocytes in vitro. This ability of the postnatal cells could be of relevance for understanding the later events in T‐cell education in the thymus and the role of I1‐2 during this process.
Detection of nephrotoxicity in cyclosporine‐A amyotrophic lateral sclerosis patients by means of urinary cytology
Authors: PECORELLA IRENE , CIARDI ANTONIO , MONGE ALESSANDRA , BRAGONI MAURA , DI TONDO UGO
Published: August 2009

Exfoliative urinary cytology was used for two amyotrophic lateral sclerosis patients treated with 3 mg/kg/day cyclosporine (CsA) therapy in order to detect the onset of the nephrotoxic side‐effects of the drug before the apparent deterioration of the patients' clinical condition. Of the two patients, only one showed clear morphological features of drug‐related damage in a one year course of cyclosporine therapy, but these followed the increase in the serum kidney and liver laboratory parameters and did not prove useful for the early detection of nephrotoxicity. However, in this patient the renal damage was hallmarked by an increasing number of tubular cells or clusters of ill‐defined renal cells in the urinary specimen, suggesting an ongoing tubular injury. The slight cytological alterations may possibly be due to the low CsA dosage used for these patients. The significance of these observations is tempered by the limited number of patients and specimens studied to date and further studies in non‐transplanted patients are therefore advocated, particularly when higher CsA doses are employed.
Murine monoclonal antibodies against pneumococcal capsular polysaccharide types 4,8,22F and 19A/19F
Authors: KOLBERG JAN , AABERGE INGEBORG S. , JANTZEN ERIK , LØVIK MARTINUS , LERMARK GRO , STEEN TORUNN
Published: August 2009

Monoclonal antibodies (MAbs) were produced against pneumococcal capsular polysaccharides after subcutaneous immunization of BALB/c mice with a 23‐valent vaccine (Pneumovax® N, Merck, Sharp & Dohme). Selected antibodies were tested in ELISA against individual polysaccharides from 23 different pneumococcal types and in a dot blot assay with heat‐killed whole bacteria adhered to nitrocellulose paper. Three MAbs (isotype IgM) were found to be specific for types 4, 8 and 22F, respectively, whereas one (isotype IgA) reacted both with 19A and 19F. Very mild acid hydrolysis of the capsular polysaccharides resulted in loss of reaction with the antibodies.
T‐cell responses in malaria
Authors: HVIID LARS , JAKOBSEN PALLE HØY , ABU‐ZEID YOUSIF ALI , THEANDER THOR G.
Published: August 2009

Malaria is caused by infection with protozoan parasites of the genus Plasmodium. It remains one of the most severe health problems in tropical regions of the world, and the rapid spread of resistance to drugs and insecticides has stimulated intensive research aimed at the development of a malaria vaccine. Despite this, no efficient operative vaccine is currently available. A large amount of information on T‐cell responses to malaria antigens has been accumulated, concerning antigens derived from all stages of the parasite life cycle. The present review summarizes some of that information, and discusses factors affecting the responses of T cells to malaria antigens.
DNA base composition, aerotolerance and enzyme patterns of Leptotrichia buccalis
Authors: HOFSTAD TOR , NÆSS VIGFRID
Published: August 2009

The guanine plus cytosine content of the DNA of Leptotrichia buccalis varied from 28.4 to 29.5 mol% (three strains). Eleven strains examined grew well under anaerobic and microaerobic conditions, but slowly in air in the presence of CO2. When examined for preformed enzymes in the APIZYM Complete Research Kit, positive reactions were obtained for several glucosidases and carboxylic ester hydrolases, and for a few peptidases.
Stereological estimation of nucleolar organizer regions in prostatic tissue with an optical disector
Authors: HANSEN ALASTAIR B. , ANDERSEN CLAUS B.
Published: August 2009

Silver‐stained nucleolar organizer regions (AgNORs) were studied in 10 hyperplastic, 10 inflamed, nine prostatic intraepithelial neoplastic (PIN) and 20 malignant prostatic lesions. Optical disector measurement on 15 urn sections showed that upper frequency limits for 1.5 um and 2.0 μm AgNOR particles were 10% and 2% in benign lesions and 18% and 9% in PIN. Using these cut‐off values for diagnosis we found a low sensitivity and high specificity in distinguishing benign lesions from PIN and PIN from carcinoma. A high sensitivity and high specificity were obtained in separating benign lesions from carcinoma. AgNOR typing on routine “two‐dimensional” sections showed that upper frequency limits for types B2, B3 and C2 were 25%, 3% and 0% in benign lesions and 38%, 23% and 11% in PIN. Using these cut‐off values for diagnosis we found lower sensitivity and specificity values among all the differential diagnostic categories. We conclude that although both methods may contribute to the differential diagnosis between benign and malignant prostatic lesions, Stereological estimation was a technically simple method, ensuring unbiased sampling and resulting in higher sensitivity and specificity. It may thus prove helpful in the differentiation of borderline cases in routine diagnostic pathology.
Redistribution of lymphocytes after cortisol administration
Authors: TOFT P. , TØNNESEN E. , SVENDSEN P. , RASMUSSEN J. W.
Published: August 2009

Major surgical procedures awake an endocrine metabolic stress response characterized by increased secretion of cortisol and lymphopenia. The purpose of this study was to clarify to which tissues the lymphocytes are redistributed after cortisol administration. Lymphocytes were isolated from 16 rabbits, labelled with indium‐111‐tropolone and reinjected into the rabbits. Eight of the rabbits received 25 mg of cortisol intravenously (group I), while eight received isotonic saline (group II). The redistribution of lymphocytes was imaged with a gamma camera and calculated by a connected computer before and two, four, and seven h after cortisol or saline administration. The radioactivity of cells from the spleen and bone marrow decreased to 84% and 56% of the initial values seven h after cortisol administration. The activity of the lymphatic tissues increased to 121% of initial values. It is concluded that during cortisol‐induced lymphopenia the lymphocytes are redistributed from peripheral blood, spleen and bone marrow to lymphatic tissue.
Induction of polyclonal antibodies to the S1 subunit of pertussis toxin by synthetic peptides coupled to PPD: Effect of conjugation method, adjuvant, priming and animal species
Authors: IBSEN P. H. , HOLM A. , RAASCHOU‐NIELSEN M. , PETERSEN J. W. , CAPIAU C. , HERON I.
Published: August 2009

Two peptides, designated L and K, covering a sequence near the NH‐terminal end of the S1 subunit of pertussis toxin (PT) were conjugated to the PPD (purified protein derivative) of M. tuberculosis by either glutaraldehyde (GLUT) or succinimidyl 4‐(N‐maleimidomethyl) cyclohexane‐1‐carboxylate (SMCC) and N‐succinimidyl 3‐(2‐pyridyldithio) propionate (SPDP) and injected into groups of mice and guinea pigs. Initially, the effect of priming the animals with BCG vaccine and the use of aluminium hydroxide as adjuvant for the anti‐peptide antibody response was studied. The group of BCG‐primed mice immunized with adsorbed peptide conjugates showed the highest anti‐peptide conjugate antibody response. Based on this finding, groups of BCG‐primed mice were immunized four times with either adsorbed peptide L‐GLUT, peptide L‐SMCC/SPDP or peptide K‐SMCC/SPDP conjugates and the fine peptide specificity as well as the PT and SI cross‐reactivity was investigated in ELISA. Mice immunized with peptide L‐GLUT showed a significant antibody response to the homologous conjugate, only, whereas the group injected with the peptide L‐SMCC/SPDP conjugate gave a significant response to both peptide K and L conjugated by the SMCC‐SPDP method. Likewise, mice immunized with the peptide K‐SMCC/SPDP conjugate reacted with the homologous and peptide L‐SMCC/SPDP conjugate, although only the response to the former conjugate was significantly greater than the response to PPD. All groups showed a strong anti‐PPD response. The anti‐PT/S1 cross‐reactivity of the antisera varied considerably within each group but was found to be highest in the peptide L‐GLUT‐immunized animals. The results of the present study not only stress the importance of BCG priming and use of aluminium hydroxide adjuvants for the immunogenicity of the peptides in question but also point to the specificity of the conjugation methods employed as low cross‐reactivity between the anti‐peptide L‐GLUT and L‐SMCC/SPDP antisera was noted. Moreover, it appeared that the choice of conjugation method may have an effect on the ability of the peptide conjugates to induce an antibody response cross‐reacting with the native protein.
Carcinoma in situ, gonadoblastoma, and early invasive neoplasia in a nine‐year‐old girl with 46, XY gonadal dysgenesis
Authors: MÜLLER JØRN , VISFELDT JAKOB , PHILIP JOHN , SKAKKEBÆK NIELS E.
Published: August 2009

Carcinoma in situ (CIS), gonadoblastoma, and early invasive neoplasia were detected in the dysgenetic gonad of a nine‐year‐old girl with 46, XY gonadal dysgenesis. A close relationship between the three neoplastic components was supported by morphological and immunohistochemical studies. Our findings support the hypothesis that all germ cell tumours, including gonadoblastomas, originate from CIS germ cells formed during early embryonic life.
Lipopolysaccharide is present in immune complexes isolated from sputum in patients with cystic fibrosis and chronic Pseudomonas aeruginosa lung infection
Authors: KRONBORG GITTE , SHAND GEOFFREY H. , FOMSGAARD ANDERS , HØIBY NIELS
Published: August 2009

Sputum samples from seven patients with cystic fibrosis and chronic P. aeruginosa lung infection were investigated for immune complexes by PEG precipitation and in two different complement binding assays. All seven patients were immune complex positive. The components involved in immune complex formation were identified by SDS‐PAGE and immunoblotting. We found P. aeruginosa lipopolysaccharide as a major antigen. Both core and O‐specific saccharide antigens could be demonstrated. IgG and IgA were the immunoglobulins involved, with IgG2 as the dominating IgG subclass. Lipopolysaccharide has a number of biological activities and its presence in sputum may have consequences for the pathogenesis of lung disease in cystic fibrosis.
Leukaemia and anaemia in AKR/O mice
Authors: HELLEBOSTAD MARIT , ØSTBYE KIRSTEN M. , HALVORSEN SVERRE
Published: August 2009

In order to study the role of the spleen in erythropoiesis during AKR/O leukaemogenesis, we have cultured bone marrow and spleen erythroid colony‐forming units (CFU‐E) and burst‐forming units (BFU‐E) from AKR/O mice (n = 40) with leukaemia of varying severity and type of manifestation. Mice with leukaemia/lymphoma had reduced concentrations of bone marrow CFU‐E and BFU‐E as compared to healthy, age‐matched AKR/O mice. The spleen content of CFU‐E was increased, and highest in mice with a spleen size between 500 and 1000 mg. The largest spleens had a somewhat lower CFU‐E content. Mice with the highest spleen CFU‐E content most often had a normal PCV; however, 4/7 had a normal bone marrow CFU‐E concentration. During AKR/O leukaemogenesis with development of spleen enlargement, the spleen may act as an erythropoietic organ, and contribute to maintaining a normal PCV. This may be a temporary ability which is reduced or lost with further progress of the disease.
Leukocyte traffic to sites of inflammation
Authors: JUTILA MARK A.
Published: August 2009

The adhesion of circulating leukocytes to the vascular endothelium is essential for effective host inflammatory and immune responses. Adhesion proteins expressed by both the leukocyte and endothelial cell have been well characterized, and studies of these molecules have shown that both cell types are actively involved in regulating these binding events. Most leukocyte (leukocyte integrins) and endothelial cell (vascular selectins, ICAM‐1, and VCAM) adhesion proteins increase in expression and function in response to mediators released by inflamed tissues. In contrast, the expression and function of one type of leukocyte molecule, L‐selectin (previously called LECAM‐1, LAM‐1, gp90MEL‐14), is “down‐regulated” by inflammatory signals. The purpose of this review is to summarize in vitro and in vivo regulatory and functional studies of some of the molecular mechanisms which regulate leukocyte‐endothelial cell adhesion, with particular emphasis on L‐selectin, and to present a hypothetical model of how these molecules may be orchestrated in vivo resulting in the control of host inflammatory responses.
Enumeration of organ‐associated natural killer cells in mice; application of a new stereological method
Authors: BASSE P. H. , HOKLAND P. , GUNDERSEN H. J. G. , HOKLAND M.
Published: August 2009

We here describe a method for estimation of the absolute number of NK cells in various organs of C57BL/6 mice. Using anti‐asialo‐GM1 heteroantisera to identify NK cells in tissue sections routinely stained by two‐layer immunoperoxidase techniques, scoring was performed using light microscopy. Extrapolation to the absolute number of NK cells/organ was done after counting 10 systematically sampled sections of each organ. Due to the thinness of the sections (8 μm) many cells are cut and will appear in at least two sections, and a correction factor was constructed to eliminate this bias. With this method we show that spleens of eight‐week‐old C57BL/6 male mice contain about 4 times 106 NK cells, which is consistent with previous findings obtained by other methods. However, application of this stereological method to liver and lungs revealed the existence of as many as 0.75 and 2.5 times 106 NK cells in these organs, respectively, i.e. three to 10 times as many as previously found in single‐cell suspensions obtained by enzymatic dissociation of liver and lung tissue. This stereological method for enumeration of the absolute number of small subpopulations of cells is generally applicable to other organs and to other antigens, and does not require the application of cumbersome single‐cell isolation procedures, during which an unknown number of cells might be lost.
Monocyte phagocytosis of opsonized Neisseria meningitidis serogroup B
Authors: SJURSEN HAAKON , LEHMANN VIDAR , NÆSS ARE , HERVIG TOR , FLØ REINHARDT W. , MÆHLE BJØRN , HALSTENSEN ALFRED I. , FRØHOLM L. ODDVAR
Published: August 2009

The chemiluminescence (CL) was examined when peripheral blood monocytes were incubated with opsonized Neisseria meningitidis, serogroup B, serotype 15:P1.16 or serotype 2a:P1.2. The monocytes were separated from a mononuclear cell suspension by an immunomagnetic negative selection technique using magnetic polystyrene microspheres coated with monoclonal antibodies specific for T and B lymphocytes. More than 90% of the lymphocytes were removed, yielding a suspension containing 93% monocytes. Optimal sensitivity for phagocytosis was obtained using 1% serum (10 μl), 72 bacteria per monocyte cell, and 7.5 min opsonization and incubation time during continuous agitation at 37°C. The CL was amplified by lucigenin. Preliminary experiments suggest that convalescent sera from patients with group B meningococcal disease induced increased CL responses compared to acute sera. Sera from volunteers immunized with an outer membrane complex vaccine from serogroup B, serotype 15:P1.16 or 2a:P1.2 meningococci also induced increased CL activity compared to preimmune sera. No such response was shown when a group B capsular polysaccharide vaccine was given. This response pattern was also demonstrated by a flow cytometric phagocytosis technique (FCM). Internalization of meningococci by monocytes was demonstrated by a FCM quenching technique and by transmission electron microscopy. CL and FCM represent rapid and reproducible methods for the measurement of opsonophagocytosis of meningococci by monocytes and may be performed with minute amounts of sera.