bluuebunny/crossref_metadata_2025 huggingface dataset:
Chlamydia pneumoniae may be associated with lung cancer:
Authors: Koyi Hirsh , Brandén Eva , Gnarpe Judy , Gnarpe HåKan , Arnholm Birgitta , Hillerdal Gunnar
Published: August 2009

Material from 117 consecutive patients with lung cancer was investigated with respect to serological markers for chronic Chlamydia pneumoniae infection. Specific C. pneumoniae IgA antibodies were found significantly more often in patients with lung cancer than in control groups with coronary heart disease and in healthy controls, even after adjustment for smoking. The results suggest that chronic C. pnmmoniae infection is common in patients with lung cancer.
Fungus‐like hyphochytrids associated with human disease
Authors: Bulajić Nina , Velimirović Slobodanka , Vukojević JElena , Nonković Zorka , Jovanović Dragutin , KuČEra IšTvan , Ilić Srbislav , BrajuŠKović Goran , Bokun Radojka , Pavlićević Goran , Trnjak Zoran
Published: August 2009

We report two cases, with liver and brain abscess, respectively, where fungus‐like organisms belonging to theHyphochytriomycotawere found at the site of inflammation together withPeptococcusin the first andCysticercus cellulosaein the second case. This is the first time these groups of organisms have been reported in human material. The role of hyphochytrids in human pathology remained uncertain as they were found together with already known human pathogens.
Radiation‐induced inflammatory malignant fibrous histiocytoma of the ileum
Authors: Fukunaga Masaharu , Endo Yasuhiko , Ushigome Shinichiro
Published: August 2009

A case of inflammatory malignant fibrous histiocytoma of the ileum seemingly induced by radiation is described. A 50‐year‐old female with a past history of uterine cervical carcinoma and postoperative radiation therapy presented with abdominal pain, fever and leukocytosis. The subserosa of the distal part of the ileum showed a diffuse dense, neutrophilic and lymphocytic infiltrate with dispersed atypical, short spindle‐ or plump oval‐shaped histiocyte‐like cells. Pleomorphic mono‐ or multinucleated giant cells with bizarre nuclei were also intermingled in the lesion. Immunohistochemically, the tumorous atypical cells were positive for vimentin, α‐smooth muscle actin, α‐1 antitrypsin and granulocyte colony‐stimulating factor. No EBV genomic sequences were detected by in situ hybridization. Flow cytometry showed an aneuploid DNA content with high S‐phase fraction. The patient was well with no evidence of tumor at 5 months after surgery. It is important to include this type of tumor in the differential diagnosis of small intestinal lesions accompanied by fever and leukocytosis following radiation.
The fluctuating pattern of various genome types of respiratory syncytial virus in Copenhagen and some other locations in Denmark
Authors: Christensen L. S. , Larsen L. B. , Johansen J. , Andersen E. A. , Wejse C. , KLUG B. , Hornsleth A.
Published: August 2009

A semi‐nested RT‐PCR method based on a region of the F and G glycoprotein genes was established, allowing the simultaneous detection and differentiation of group A and group B isolates of respiratory syncytial virus (RSV). The PCR products were subjected to digestion with restriction endonucleases to further differentiate the isolates. Using, in addition, previously reported studies the prevalence of various genome types in the Copenhagen region over a period of 6 years was established. Furthermore, the prevalence of genome types was determined in a distant region in Denmark during the winters of 1996/97 and 1997/98, and in yet another distant region during the winter of 1997/98. It was shown that the different regions in Denmark to a large extent share the same pool of genome types of RSV. Yet, while the fluctuating patterns of the two groups and various genome types were almost identical at different hospitals in the Copenhagen region, they varied between the different regions. This suggests that epidemics in local communities primarily rely on region‐specific herd immunity parameters and emerge from strains endemically circulating in these local communities. Group B strains in Copenhagen showed an overall predominance, being predominant in three of the six epidemic seasons studied, and of almost equal predominance in one season.
Multidrug resistance and retroviral transduction potential in human small cell lung cancer cell lines
Authors: Theilade Mette Due , Gram Gregers Jacob , Jensen Peter Buhl , Cianfriglia Maurizio , RøRth Mikael , Hansen John‐Erik Stig
Published: August 2009

Multidrug resistance (MDR) remains a major problem in the successful treatment of small cell lung cancer (SCLC). New treatment strategies are needed, such as gene therapy specifically targeting the MDR cells in the tumor. Retroviral LacZ gene‐containing vectors that were either pseudotyped for the gibbon ape leukemia virus (GALV‐1) receptor or had specificity for the amphotropic murine leukemia virus (MLV‐A) receptor were used for transduction of five SCLC cell lines differing by a range of MDR mechanisms. Transduction efficiencies in these cell lines were compared by calculating the percentage of blue colonies after X‐Gal staining of the cells grown in soft agar. All examined SCLC cell lines were transducible with either vector. Transduction efficiencies varied from 5.7% to 33.5% independent of the presence of MDR. These results indicate that MDR does not severely impair transduction of SCLC cells, and that MLV‐A as well as GALV‐1 retroviral vectors are suitable for further development of gene therapy in SCLC.
The use of CHROMagar Orientation as a primary isolation medium with presumptive identification for the routine screening of urine specimens
Authors: Houang Elizabeth T. S. , Tam Po Chu , Lui Sau Lai , Cheng A. F. B.
Published: August 2009

The aim of the study was to compare the use of a novel differential culture medium CHROMagar, for both primary isolation and presumptive identificaton, with the method currently used in our laboratory for screening mid‐stream‐urine samples (MSU). Routine methods (RM) included blotting paper imprinting of all specimens and additional quantitative culture on cysteine lactose electrolyte‐deficient agar (CLED) for selected samples together with Microbact 12E for further identification. The CHROMagar method (CH) relied on the use of blotting paper imprints, colonial colour and morphology on CHROMagar only. With respect to the 3390 MSU specimens examined, both methods yielded similar results in 3240, including ≤87% of Escherichia coli, Pseudomonas spp., Staphylococcus spp., Proteus mirabilis/ Morganella morganii and Enterobacter/Serratia/Klebsiella/Citrobacter spp. Of the 52 discordant identifications, yeasts were reported as staphylococci on CHROMagar in 10. The overall cost of materials per specimen was US$ 0.30 by RM and $ 0.24 by CH. It took about 3 min to perform each Microbact test. Thus, CHROMagar plus Gram stain and other simple bench tests gave results similar to those using our current method, but had the advantage of saving time and materials.
A histomorphometric method for evaluating topographic differences in degree of early atherosclerosis within the human aorta
Authors: Ageræk Mads , Kristensen Ingrid B. , Pedersen Erik M.
Published: August 2009

The aim of the study was to develop an unbiased topographically oriented method of evaluating early atherosclerotic lesions in the aorta and to apply this to a series of human aortas from young adults. A systematic sampling procedure and histomorphometric analysis of intimal thickening is described. Results from a group of 15 young adults (aged 18–40 years) showed a characteristic pattern with increasing intimal thickening when moving distally from the thoracic to the upper and finally to the lower abdominal aorta, but also a shift in the localization of the most pronounced intimal thickening from the posterior to the anterior and back to the posterior aspect. This pattern was found in aortas both with minimal and with more pronounced atherosclerosis, and supports the view that the early intimal thickening precedes the atherosclerotic lesions and marks the sites of predilection for the more advanced disease processes. An increase in intimal thickness with age could be demonstrated in the aortas without overt atherosclerosis. The simple sampling procedure, well‐defined sampling sites, and ability to demonstrate and quantitate differences in intimal thickening and plaque morphology make this method well suited for relating morphometric data to other parameters of interest when studying the etiology and dynamics of atherosclerotic disease.
Properties and distribution of the putative R3 protein of Streptococcus agalactiae
Authors: Kvam A. I. , Bevanger L. , Mæland J. A.
Published: August 2009

Strain‐variable Streptococcus agalactiae (group B streptococci; GBS) proteins exposed at the bacterial cell surface are important markers in GBS serotyping. These proteins include the c proteins cα and cβ and the R proteins R1 through R4, of which R1 and R4 have been studied most extensively. This study presents the characteristics of a protein which was expressed by a capsular antigen type V GBS strain shown by means of polyclonal and monoclonal antibody testing. Examination of a number of reference and prototype strains by fluorescent antibody testing and Western blotting provided evidence that the serotype V‐derived protein was the R3 protein of GBS, previously defined on the basis of immunoprecipitation assays. The putative R3 protein formed ladder‐like banding patterns on Western blotting with polypeptides in the 30 kDa to ≥ 140 kDa range, was destroyed by pepsin digestion, and partially degraded by trypsin digestion. The protein was expressed by 10 (6.5%) of 153 clinical GBS strains tested, the expression being restricted to isolates of the capsular antigen types II, III, and V. Some isolates expressed both the cβ and the R3 protein. Expression in combination with cα or R4 protein synthesis was not detected. Inclusion of the anti‐R3 monoclonal antibody among antibody reagents for GBS serotyping will enhance the discriminatory power of this typing method.
Expression of in vitro virulence by Entamoeba histolytica: effect of calmodulin inhibitors
Authors: Arias‐Negrete Sergio , Munoz Mari̇a De Lourdes , Murillo‐Jasso Faustino
Published: August 2009

The possible role of calmodulin (CaM) in functions such as cytotoxicity, phagocytosis, and amoebic growth was studied. These are important factors for the in vitro pathogenicity of Entamoeba histolytica HM1 strain. The CaM inhibitors trifluoperazine (TFP), N‐(6‐aminohexyl)‐5‐chloro‐1‐naphtha‐lene‐sulfonamide (W‐7) and N‐(6‐aminohexyl)‐naphthalenesulfonamide (W‐5) were used for these purposes. Cytotoxicity was determined as the ability of amoebae to kill and/or lyse nucleated mouse spleen cells. Phagocytosis by amoebae was assessed by calculating the number of endocytosed human red blood cells. Cytotoxicity by E. histolytica showed a decrease dependent upon the concentration of W‐7 and TFP, whereas W‐5 did not show a significant inhibiting effect. Phagocytosis was roughly sensitive to W‐7; TFP and W‐5 did not have any significant effect. Amoebic growth was sensitive to TFP and W‐7 CaM inhibitors. These results suggest that CaM participates in the expression of in vitro cytotoxicity of E. histolytica.
Demonstration of Chlamydia pneumoniae in tissue by immunohistochemistry
Authors: NääS Jessica , Gnarpe Judy A.
Published: August 2009

Chronic Chlamydia pneumoniae infection may be difficult to diagnose using routine methods such as culture, antigen detection, serology and polymerase chain reaction (PCR). We describe a method for the immunohistochemical staining of tissue biopsies using a C. pneumoniae‐specific monoclonal antibody and a streptavidin/biotin technique. This method provides specific identification of the organism and reveals its actual location in tissues which may or may not be inflamed. Host cells containing C. pneumoniae antigen were easily identified by their bright red color in tissue sections counterstained with hematoxylin.
Signal transduction by the major histocompatibility complex class I molecule:
Authors: Pedersen Anders E. , Skov SøRen , Bregenholt SøRen , Ruhwald Morten , Claesson Mogens H.
Published: August 2009

Ligation of cell surface major histocompatibility class I (MHC‐I) proteins by antibodies, or by their native counter receptor, the CD8 molecule, mediates transduction of signals into the cells. MHC‐I‐mediated signaling can lead to both increased and decreased activity of the MHC‐I‐expressing cell depending on the fine specificity of the anti‐MHC‐I antibodies, the context of CD8 ligation, the nature and cell cycle state of the MHC‐I‐expressing cell and the presence or absence of additional cellular or humoral stimulation. This paper reviews the biochemical, physiological and cellular events immediately after and at later intervals following MHC‐I ligation. It is hypothesized that MHC‐I expression, both ontogenically and in evolution, is driven by a cell‐mediated selection pressure advantageous to the MHC‐I‐expressing cell. Accordingly, in addition to their role in T‐cell selection and functioning, MHC‐I molecules might be of importance for the maintenance of cellular homeostasis not only within the immune system, but also in the interplay between the immune system and other organ systems.
Pathophysiological effects of human TNF‐alpha‐producing tumor xenografts in immunosuppressed mice
Authors: Nagy T. , J´nossy T. , Vizler C. , Bohus K. , Jóa F. , Végh P. , Duda E.
Published: August 2009

Groups of CBA mice immunosuppressed with antithymocyte serum (ATS) treatment were xeno‐transplanted with either HeLa human cervical carcinoma cells or genetically modified cells expressing the human tumor necrosis factor‐α (TNF) gene (All cells). Both cell lines were highly resistant to the cytotoxic effects of TNF. If 3 times 106 tumor cells were inoculated s.c. into female mice, HeLa cells grew progressively into large tumors and killed 74% of the recipients, while TNF‐expressing All cells caused fatal tumor growth only in 22% of the mice. 3times 106 or 1.5times 107 All cells produced progressive tumor growth and lethality in all male recipients. In sera of all the All‐cell‐transplanted mice, biologically active TNF was detected shortly (4.5 h) after tumor inoculation (6–39 U/ml), decreasing to below detection level in the circulation by day 3. In recipients of 15 million All cells, circulating TNF reappeared and reached high levels (12–1000 U/ml) 3 to 7 weeks later, when the animals bore large tumors (14–23 mm). Generally, such mice became cachectic, severely anemic, hypothermic, and soon died. On account of calcium mobilization from bones, their serum Ca levels were high. Electron microscopy revealed severe liver damage, but there were no signs of chronic arthritis. These results suggest that ATS‐treated mice xenotransplanted with TNF‐gene‐transfected All human tumor cells provide a new model for studying the pathophysiological and anti‐tumor effects of TNF.
Experimental corticosteroid induction of Pneumocystis carinii pneumonia in piglets
Authors: Nielsen Jens , Bille‐Hansen Vivi , Settnes Osvald Peter
Published: August 2009

Animal models of Pneumocystis carinii (Pc) pneumonia (PCP) play a central role in research on the Pc microorganism itself and the disease, especially the pathogenesis and the host defence. The classic rat model with corticosteroid‐induced reactivation of a latent infection has been most widely used. In our search for alternative non‐rodent models, six 31/2‐week‐old piglets were injected intramuscularly with methylprednisolone acetate, at 18 mg/kg body weight, once a week for 6 weeks. Six littermate piglets constituted the control group. The principals showed a markedly lower growth rate than the controls. Furthermore, they developed “moon face” and “pot belly”, snoring sounds while eating, and pronounced respiratory distress during handling. Significant changes in haematological parameters, including lymphopenia, were observed in the principal group. The Pc antibody titres of the controls increased to high levels, whereas the principals were all low‐titred or seronegative for Pc at the last blood sampling. At necropsy, the mean body weight of the principals was about half that of the controls. In addition, they had an extreme reduction of the thymus together with dark red consolidations of the frontal lung lobes and/or atelectatic looking diaphragmatic lobes. Histopathologically, there was a focal interstitial pneumonia. Alveolar walls and interstitia had mononuclear cell infiltrations and the alveolar lumina were occluded by foamy acidophilic honeycomb material with a varying number of Pc cysts. The reduced body weight, the thymus involution, and the lymphopenia, together with the reduced levels of specific Pc antibodies and the histomorphology of the PCP, were consistent parameters of the principal group and comparable to the findings of the classic rat model. Thus, the present study is the first to describe that prolonged administration of high doses of methylprednisolone acetate can induce PCP in piglets.
Migration of neutrophils is dependent on mast cells in nonspecific pleurisy in rats
Authors: Nishida Minoru , Uchikawa Ryuichi , Tegoshi Tatsuya , Yamada Minoru , Matsuda Shinji , Hyoh Yukihiko , Arizono Naoki
Published: August 2009

To determine the role of mast cells in the recruitment of neutrophils and eosinophils, acute nonspecific pleurisy was induced by injecting isologous serum into normal +/+ and mast cell‐deficient Ws/Ws rats. In +/+ rats, neutrophil infiltration peaked 4 h after serum administration, followed by influx of eosinophils after 24–48 h. The levels of neutrophil influx after 4 h as well as the activity of myeloperoxidase (MPO) in pleural lavage‐cell extract were significantly lower in Ws/Ws rats than in +/+ rats. In contrast, numbers of eosinophils as well as activity of eosinophil peroxidase (EPO) did not differ significantly between Ws/Ws and +/+ rats. For local reconstitution of mast cells, +/+ rat peritoneal mast cells (PMC) or mesenteric lymph node cells (MLNC) as a control were transferred into the Ws/Ws pleural cavity. Serum injection into animals with PMC transfer 7 days previously triggered augmented neutrophil influx by approximately 4.7‐fold as compared to that in MLNC‐transferred animals. Mast cells recovered from the pleural cavity of PMC‐transferred rats showed histamine contents equivalent to 20% of that of freshly isolated PMC and retained the reactivity to compound 48/80. These results indicated that dependency of neutrophil recruitment on resident mast cells is greater than that of eosinophils in isologous serum‐induced pleurisy.
Histopathological findings in renal allografts at time of transplantation and correlation with onset of graft function*
Authors: Lehtonen Satu R. K. , Taskinen Eero I. , Isoniemi Helena M.
Published: August 2009

Histopathological changes quantified using the chronic allograft damage index (CADI) have been shown to predict subsequent graft outcome and developing chronic rejection. The aim of the study reported here was to investigate the extent to which cadaveric renal allografts exhibit histopathological changes at time of transplantation, focusing on changes covered by the CADI. We also analysed whether any histopathological change predicts delayed graft function. One hundred and twenty‐eight cadaveric kidney allografts with adequate protocol biopsy were studied. Tubular epithelial anisometric vacuolization was the commonest change, found in 62% of grafts and scored moderate or severe in 28% of these cases. Other prevalent changes were interstitial fibrosis, vascular hyalinosis, glomerular sclerosis and tubular basement‐membrane thickening in 40%, 37%, 28% and 22% of biopsies, respectively. Intensities were, however, scored as mild in over 95% of specimens. The mean CADI for all grafts was 0.74. A significant difference in CADIs was seen between grafts from donors under and over 40 years of age. Grafts with early, delayed and no function had a similar incidence of histopathological changes. Histopathological changes in renal allografts were mostly uncommon and mild at time of transplantation, but some grafts exhibited changes which were quantified using the CADI. Though histopathological changes quantified with the CADI are predictive of subsequent graft function, they did not affect onset of graft function.
Bamboo joint‐like appearance of the stomach: a histopathological study
Authors: Hirokawa Mitsuyoshi , Shimizu Michio , Terayama Kiyomi , Tamai Minako , Takeda Masaharu , Iida Mitsuo , Manabe Toshiaki
Published: August 2009

A bamboo joint‐like (BJL) appearance is an endoscopic finding characterized by swollen longitudinal folds transversed by erosive fissures or linear furrows, and it seems to be associated with Crohn's disease (CD). We performed gastroduodenoscopy in 1781 patients, and found BJL lesions in 15 (65.2%) of 23 CD patients and 20 (1.1%) of 1758 non‐CD patients. Histological examination of the biopsy specimens showed sharp, fissure‐like erosion or mucosal cleft in 7 (50%) out of 14 CD patients and one (20%) out of 5 non‐CD patients. All cases with fissure‐like erosion or, mucosal cleft revealed lymphoid aggregates, eosinophilic infiltration and edema in the superficial portion of the surrounding lamina propria. Epithelioid granuloma was seen at the base of the fissure‐like erosion in two cases with CD. Our results indicated that fissure‐like erosion or mucosal cleft in gastric biopsy specimens reflects the BJL appearance, and that such findings may provide a diagnostic clue to CD.
The 1998 list Proposed new bacterial taxa and proposed changes of bacterial names published during 1998 and considered to be of interest to medical or veterinary bacteriology:
Authors: Frederiksen Wilhelm , Magee John , Ursing JAn
Published: August 2009

A list of names published or validated in 1998 is presented.
Lactoferrin: A multifunctional glycoprotein
Authors: Vorland Lars H.
Published: August 2009

Lactoferrin is an iron‐binding glycoprotein found in milk, exocrine secretions of mammals, and in secondary granules from polymorphonuclear neutrophils. This review describes the wide spectrum of functions ascribed to lactoferrin, with special emphasis on the antimicrobial properties of this protein, and its derived peptides.
High antibiotic consumption in Danish intensive care units?
Authors: Petersen In A Sleimann , Hesselbjerg Lars , JøRgensen Lise , Renstrur Jens , Barnung Steen , Schierbeck Jens , Jepsen Ole Bent
Published: August 2009

Decreased antibiotic susceptibility among microorganisms isolated from intensive care unit (ICU) patients is found to be associated with high total antibiotic consumption or inappropriate use of antibiotics in the ICUs. The aims of this study were: 1) to characterize the antibiotic consumption in Danish ICUs, and in four ICUs with expectedly large differences in levels of antibiotic consumption, 2) to estimate the association between antibiotic susceptibility among isolated microorganisms and antibiotic consumption. This was done by: 1) a retrospective questionnaire study of the annual supply of antibiotics in 1995 to 30 ICUs in Denmark, and 2) a 2‐month prospective study of patients and microbiological samples in four Danish ICUs in 1996. We found that the supply of antibiotics to Danish ICUs was substantial, with a median value of 124 DDD/100 patient days. No association was found between high consumption of antibiotics and decreased antibiotic susceptibility in the four ICUs.
The terminal complement complex is generated in chronic leg ulcers in the absence of protectin (CD59)
Authors: Balslev Eva , Thomsen Henrik Klem , Danielsen Lis , Sheller Jeanet , Garred Peter
Published: August 2009

Loss of membrane complement regulators accompanied by complement activation is suggested to be involved in the pathophysiological processes leading to tissue damage in myocardial ischaemia. In the present study we have investigated whether the same phenomenon may occur in ischaemic and/or venous hypertension leg ulcers. The deposition of complement, plasma complement regulators and expression of membrane regulators were detected by immunohistochemical methods, including immunofluorescence with antibodies against C3d, the terminal complement complex (TCC), vitronectin, clusterin, decay‐accelerating factor (CD55) and protectin (CD59). Eleven frozen biopsies from ischaemic leg ulcers, 10 biopsies from venous hypertension leg ulcers, and 10 biopsies from normal skin were studied. In 9 of 11 ischaemic and in 5 of 10 venous hypertension leg ulcers, marked staining for TCC was found around the capillaries, most often at the ulcer margin. No TCC staining was found in normal skin. Staining for TCC was always accompanied by staining for clusterin and vitronectin and C3d. In normal skin, CD59 was found on the elastic fibers in the dermis, on the muscle coat, the Schwann sheath and acinar cells. Semiquantitative measurement of CD59 showed marked increased staining intensity in the endothelium in venous hypertension ulcers and diminished intensity in ischaemic ulcers compared to normal skin. No such difference could be observed for CD55. When TCC was positive in the capillary walls, weak or no staining for CD59 was found. A significantly higher ratio of TCC/CD59 was found in the ischaemic compared to venous ulcers (p=0.018). This was due to a marked difference between the ulcer margins (p=0.013). Localized areas in the venous ulcers had the same pattern as that seen in the ischaemic ulcers. Our results suggest that loss of CD59 may enhance deposition of TCC and that complement‐dependent inflammation may be an important factor in the tissue‐damaging processes seen in chronic leg ulcers.
In situ localization of S‐phase‐specific histone (H3) mRNA in Bowen's disease
Authors: Murakami Masamoto , Mizoguchi Yoshikazu , Horibe Yoshimune , Komori Katsuhiko , Hori Haruna , Kasahara Masao
Published: August 2009

PCNA and Ki‐67 immunohistochemistry has been used to assess cell proliferation in place of tritiated thymidine or BrdU labeling of S‐phase cells. Recently, it has been possible to reliably demonstrate histone H3 mRNA by in situ hybridization in formalin‐fixed and paraffin‐embedded tissue sections. We have compared this new proliferation marker with Ki‐67 and PCNA with regard to distribution of positive cells and labeling indices (LI%) for 22 cases of Bowen's disease.

In normal skin, Ki‐67‐IHC positive cells and histone mRNA positive cells were observed in the basal and suprabasal layers of the epidermis. In Bowen's disease, positive cells with each marker were more frequent in upper neoplastic epidermis than in suprabasal layers, and the average LI%s were markedly elevated with all markers, the scores decreasing in the following order: PCNA‐IHC, Ki‐67‐IHC and H3mRNA‐ISH. However, the results of double staining demonstrated that S‐phase cells do not necessarily show exactly the same distributions as with PCNA and Ki‐67‐IHC labeling. H3mRNA‐ISH showed three different degrees of reaction with significantly different LI%s, whereas PCNA and Ki‐67 LI% did not vary essentially in the same areas. These results strongly suggest that Bowen's disease, which is well known as a low‐grade neoplastic state with malignant potential, also demonstrates clear intratumoral heterogeneity of S‐phase cells using the H3mRNA‐ISH method.
Functional differentiation of acute myeloid leukaemia blast cells
Authors: BassøE Carl‐Fredrik , Halstensen Alfred , BRUSERUD øYSTEIN
Published: August 2009

Little is known of the functional status of blast cells from patients with acute myeloid leukaemia (AML). We have studied phagocytosis and membrane receptors by flow cytometry (FCM), and secretory activities in blast cells from 24 AML patients prior to treatment. Blast cells from 11/16 patients attached N. meningitidis, and internalization occurred in 7/14. The phagocytosis of zymosan particles and N. meningitidis correlated linearly (r=0.9, p<0.01, n=11). Surface membrane expression of CD32 and CD1 1b was sufficient to account for opsonin‐dependent attachment in all except one patient. A significant fraction of the blast cells attached, but did not internalize meningococci. CD32 and CD11b were non‐functional in all the blasts from five patients, and in a subpopulation from seven additional patients. Significantly more large than small blasts expressed CD32, CD35 and CD11b (p<0.001). Phagocytosis was unrelated to the secretion of IL‐1α, IL‐1β, and TNFα. In conclusion, AML blast cell function is related to receptor expression, cell size and granularity, and to FAB‐type.
The relationship between the acid and alkaline phosphatase activity and the adherence of clinical isolates of Candida parapsilosis to human buccal epithelial cells
Authors: Fernanado P. H. R. , Panagoda G. J. , Samaranayake L. P.
Published: August 2009

Candida parapsilosis is an emerging fungal pathogen implicated in many diseases, especially in compromised hosts. Candidal colonization and infection depends on the initial ability to adhere to host surfaces, which in turn depends upon the cell wall components and the allied structures of both the host and the fungus. Examination of a miscellaneous collection of 24 C. parapsilosis isolates, from both superficial and deep infections, for their potential pathogenic traits displayed a relationship between the phosphatase activity measured with p‐nitrophenol phosphate and adhesion of the yeasts to human buccal epithelial cells (BECs). Significant intraspecies differences were seen in both the alkaline and acid phosphatase activity as well as in their adhesion to BECs (p<0.0001). The acid phosphatase activity of the superficial isolates was significantly greater (152%) than that of the systemic isolates (p=0.0352). A highly significant positive correlation was also established between the yeast adhesion to BECs and both the acid (r=0.88, p<0.0001) and alkaline (r=0.9, p<0.0001) phosphatase activity. These relationships, described here for the first time, imply that phosphatases of Candida species may play a crucial role in potentiating their virulence.
Apoptosis and expression of caspases 3,6 and 8 in malignant non‐Hodgkin's lymphomas
Authors: Soini Ylermi , PääKKö Paavo
Published: August 2009

In this study we investigated the immunohistochemical expression of caspases 3, 6 and 8 in 85 malignant non‐Hodgkin's lymphomas and in 4 hyperplastic lymph nodes. The extent of apoptosis and the immunohistochemical expression of bcl‐2 and bax was also studied. Caspase 3 immunoreactivity was seen in 84/85 (99%), caspase 6 in 46/85 (54%), and caspase 8 in 66/85 (78%) lymphomas. The immunoreactivity for caspase 3 was diffuse cytoplasmic while antibodies to caspase 6 and 8 showed granular and fragmented, sometimes also nuclear immunopositivity. High‐grade non‐Hodgkin's lymphomas expressed strong caspase 6 and 8 immunoreactivity significantly more often than low‐grade lymphomas (p=0.016 and p=0.0002, respectively). Strong caspase 3 immunoreactivity was also seen more often in high‐grade lymphomas, but the association did not reach statistical significance (p=0.14). There was a strong association between the expression of caspase 3 and 6 (p=0.032), caspase 3 and 8 (p=0.042), and especially between caspase 6 and 8 (p=<0.00001). There was a significant difference in the apoptotic index between low‐grade (0.59±0.44%) and high‐grade lymphomas (1.96±1.92%) (p<0.001). Strong bcl‐2 expression was seen in 35/80 (44%) and strong bax expression in 20/80 (25%) lymphomas. No significant association was found between the expression of bcl‐2 or bax and the expression of the caspases. According to the results the expression of caspases 6 and 8 is upregulated in high‐grade compared with low‐grade lymphomas and probably contributes to the execution of apoptosis in them. A similar tendency could also be seen with caspase 3. The expression of the three caspases is significantly associated, suggesting that it is mutually regulated. Finally, the results suggest that the expression of bcl‐2 or bax does not influence the expression of caspases 3, 6 and 8 in malignant lymphomas to a significant degree.
Is streptolysin S of group A streptococci a virulence factor?
Authors: Ginsburg Isaac
Published: August 2009

The possible role played by streptolysin S (SLS) of group A streptococci in the pathophysiology of streptococcal infections and in post‐streptococcal sequelae is discussed. The following properties of SLS justify its definition as a distinct virulence factor: 1) its presence on the streptococcus surface in a cell‐bound form, 2) its continuous and prolonged synthesis by resting streptococci, 3) its non‐immunogenicity, 4) its extractability by serum proteins (albumin, alpha lipoprotein), 5) its ability to become transferred directly to target cells while being protected from inhibitory agents in the milieu of inflammation, 6) its ability to bore holes in the membrane phospholipids in a large variety of mammalian cells, 7) its ability to synergize with oxidants, proteolytic enzymes, and with additional host‐derived proinflammatory agonists, and 8) its absence in streptococcal mutants associated with a lower pathogenicity for animals. Because tissue damage in streptococcal and post‐streptococcal sequelae might be the end result of a distinct synergism between streptococcal and host‐derived proinflammatory agonists it is proposed that only cocktails of anti‐inflammatory agents including distinct inhibitors of SLS (phospholipids), gamma globulin, inhibitors of reactive oxygen species, proteinases, cationic proteins cytokines etc., will be effective in inhibiting the multiple synergistic interactions which lead to fasciitis, myositis and the flesh‐eating syndromes, and often develop into sepsis, septic shock and multiple organ failure. The creation of mutants deficient in SLS and in proteases will help shed light on the specific role played by SLS in the virulence of group A hemolytic streptococci.
Serological response to Helicobacter pylori recombinant antigens in Chilean infected patients with duodenal ulcer, non‐ulcer dyspepsia and gastric cancer
Authors: Opazo Patricio , Müller Ilse , Roll´n Antonio , Valenzuela Pablo , Yudelevich Arturo , De La Guarda Ruth García‐ , Urra Soledad , Venegas Alejandro
Published: August 2009

We have previously cloned 10 Helicobacter pylori antigen genes from a Chilean strain including: cytotoxin VacA, a truncated region of CagA (called A17), a species‐specific protein (Ag26), urease subunits (UreA, UreB), a flagellin, (FlaB), heat shock proteins (HspA and HspB), an adhesin (HpaA) and a lipoprotein (Lpp20). Immunogenicity of these antigens was tested by immunoblot with sera of Chilean infected patients, revealing that HpaA, A17, HspB and VacA were more frequently recognized (86%, 82%, 68% and 68%, respectively). According to the clinical condition, it was determined that Lpp20 was preferentially recognized by sera from non‐ulcer dyspepsia patients (80%), A17 and VacA by patients with duodenal ulcer (92% and 83% respectively), and HspB by patients with duodenal ulcer (83%) and gastric cancer (90%). An ELISA was developed with a purified mixture of A17 and VacA antigens to test the different groups of patients. It was found that sera from duodenal ulcer patients showed higher values than those from non‐ulcer dyspepsia patients, but this difference was not significant (p<0.2). Moreover, sera from gastric cancer patients showed values lower than those from non‐ulcer dyspepsia patients (p<0.019). These results indicate that, in the Chilean population, antibodies raised against VacA and A17 are not markers either for duodenal ulcer or for gastric cancer.
The discriminatory power of ribo‐PCR compared to conventional ribotyping for epidemiological purposes
Authors: Severino Patrícia , Darini Ana Lúcia , MagalhäEs Vanda D.
Published: August 2009

Molecular typing techniques have become increasingly important for confirmation of epidemiological relationships and delimitation of nosocomial outbreaks. The discriminatory power of the two DNA‐based typing methods, conventional ribotyping and ribo‐PCR, was assessed to distinguish between selected strains of Acinetobacter calcoaceticus, Enterobacter cloacae, Serratia marcescens and Pseudomonas aeruginosa. Overall, conventional ribotyping was more discriminatory than ribo‐PCR.
Low occurrence of antibiotic resistance in Escherichia coli and staphylococci isolated from blood cultures in two Norwegian hospitals in 1991–92 and 1995–96
Authors: Leegaard Truls M. , Vik Einar , Caugant Dominique A. , FrøHolm L. Oddvar , HøIby E. Arne
Published: August 2009

The aim of this study was to investigate the antibiotic resistance rates of major bacterial pathogens causing bloodstream infections in two very different types of hospital in Norway. We examined all Escherichia coli and staphylococci (330 isolates) causing bloodstream infections from one general county hospital and one specialist national cancer hospital during the periods 1991–92 and 1995–96. Minimal inhibitory concentrations (MICs) were determined using the E‐test. E. coli and staphylococci constituted 46.7% of all isolates from bloodstream infections in the two hospitals. Overall, E. coli isolates were resistant to amoxicillin (21%), trimethoprim (21%), doxycycline (20%) and trimethoprim‐sulphamethoxazole (17%), while Staphylococcus aureus strains were resistant to benzylpenicillin (66%). No methicillin‐resistant S. aureus was detected. Coagulase‐negative staphylococci were often multires‐istant, but remained fully sensitive to vancomycin. For a few antibiotics, significantly more resistance was found in the specialist hospital. In our material we found no significant increase in resistance between 1991–92 and 1995–96. In conclusion, antimicrobial resistance still remains low in important bacterial pathogens causing bloodstream infections in Norway.
Early immune response in susceptible and resistant mice strains with chronic Pseudomonas aeruginosa lung infection determines the type of T‐helper cell response
Authors: Moser Claus , Hougen Hans Petter , Song Zhijun , Rygaard JøRgen , Kharazmi Arsalan , HøIby Niels
Published: August 2009

Most cystic fibrosis (CF) patients become chronically infected with Pseudomonas aeruginosa in the lungs. The infection is characterized by a pronounced antibody response and a persistant inflammation dominated by polymorphonuclear neutrophils. Moreover a high antibody response correlates with a poor prognosis. We speculated that a change from this Th2‐like response to a Th1‐like response might decrease the lung inflammation and thus improve the prognosis in CF patients. To investigate this, we infected C3H/HeN and BALB/c mice intratracheally with P. aeruginosa. In addition, we studied the early immune response leading to different Th responses. Mortality was lower in the C3H/HeN mice (p<0.005), they cleared the bacteria faster (day 3 p<0.01, day 7 p<0.02), had a milder lung inflammation (day 7 p<0.01, day 14 p<0.0005) and had a Th1‐like IgG subclass switch. At day 3, the C3H/HeN mice produced less NO and TNF‐α, (p<0.01 and p<0.03) and had the lowest IL‐10/IL12 ratio (p<0.05). At day 7, the C3H/HeN mice had the highest IFN‐γ (p<0.02), and the lowest IL‐4 (p<0.02) production in the lungs. In conclusion, these results show that the Th1‐reacting C3H/ HeN mice with chronic P. aeruginosa lung infection have a better disease outcome compared to the Th2‐reacting BALB/c mice, indicating that a Th1 response might be beneficial in CF patients with chronic P. aeruginosa lung infection.
Immunohistochemical localization of FAP‐1, an inhibitor of Fas‐mediated apoptosis, in normal and neoplastic human tissues
Authors: Lee Sug Hyung , Shin Min Sun , Park Won Sang , Kim Su Young , Kim Hong Sug , Lee Jong Heun , Han Seo Young , Lee Hun Kyung , Park Jik Young , Oh Ro Ra , Jang Ja June , Lee Jung Young , Yoo Nam Jin
Published: August 2009

Fas, a death receptor, is widely expressed in human tissue, but its expression, although a prerequisite for the induction of apoptosis, does not predict its biological function. To understand the mechanisms of Fas resistance in human tissues in vivo, we performed immunohistochemistry using an antibody against Fas‐associated phosphatase‐1 (FAP‐1), which interacts with the cytosolic domain of Fas and inhibits Fas‐mediated apoptosis. In normal human tissues, FAP‐1 immunostaining was easily detected, for example, in renal tubules, skeletal muscle, myocardiocytes, pituitary gland, parathyroid gland, pancreatic islets, hepatocytes, testicular germ cells, prostatic glands, neurons, epithelium of fallopian tube, endometrial glands, trophoblasts, bronchial epithelial cells, and some types of gastrointestinal epithelial cells. In 123 (78%) of 158 cancers of various origins, including breast carcinomas, stomach carcinomas, colon carcinomas, lung carcinomas and several types of sarcomas, variable intensities of FAP‐1 expression were evident. Taken together, these findings demonstrated that FAP‐1 is widely expressed in normal human tissues and partly overlapped with Fas expression described in earlier reports, suggesting that FAP‐1 may have an important role in the regulation of apoptosis in vivo. In addition, FAP‐1 expression in cancers suggests that many cancers may be resistant to Fas‐mediated apoptosis through the action of FAP‐1 in vivo.
Rapid identification of mutations in a multidrug efflux pump in Pseudomonas aeruginosa
Authors: Jalal Shah , Wretlind Gunnar , Gotoh Naomasa , Wretlind Bengt
Published: August 2009

The gene mexR regulates negatively the expression of the MexA‐MexB‐OprM efflux pump in Pseudomonas aeruginosa, and mutations in mexR cause a multiple antibiotic resistance phenotype. Five hundred and forty resistant clones of P. aeruginosa PAO503 were isolated after selection for resistance to chloramphenicol or tetracycline. All isolates showed similar phenotypes and were resistant to tetracycline, chloramphenicol and norfloxacin. Nineteen randomly selected isolates were analyzed. Since mutational analysis by direct sequencing of all regions of interest in several strains is time‐consuming and expensive, a screening method, Non‐Isotopic RNase Cleavage Assay (NIRCATM), was applied to identify mutant genes so that they could be targeted for DNA sequencing. NIRCA is a simple but rapid method for mutational analysis and can be performed in 3–4 h. Results of NIRCA analysis were compared with DNA sequencing. Both NIRCA and DNA sequencing analysis showed mexR gene mutations in 11 of 19 isolates but no alterations in 8 strains. An immunoblot assay showed overexpression of OprN, a component of another multidrug efflux pump, MexE‐MexF‐OprN, in those eight isolates. Nucleotide sequencing of quinolone resistance‐determining regions of DNA gyrase (gyrA) or topoisomerase IV (parC) showed no alterations in any of the 19 mutants. The data indicate that two efflux pump systems, MexA‐MexB‐OprM and MexE‐MexF‐OprN, were involved in multidrug resistance including quinolones and that NIRCA is a sensitive method for screening mutations.
Proinflammatory activation of neutrophils and monocytes byHelicobacter pyloriis not associated withcagA,vacA orpicB genotypes
Authors: Hansen Per Syrak , Go Mae F. , Varming Kim , Andersen Leif P. , Graham David Y. , Nielsen Henrik
Published: August 2009

ChronicHelicobacter pyloriinfection is associated with mucosal inflammation. The aim of the present study was to assess human neutrophil and monocyte activation induced byH pyloristrains with different virulence genotypes. Bacterial sonicates from 12 strains were used to induce phagocyte up‐regulation of adherence molecule CD11b, assessed by fluorescence flow cytometry, and oxidative burst responses, assessed by chemiluminescence. A dose‐dependent induction of the expression of CD11b was observed with sonicate from allH. pyloristrains on both neutrophils and monocytes. Strains negative forcagA andpicB genes had the same inducing activity of upregulation of CD11b as strains positive for these genes. AvacA‐S2 type strain had the same activity asvacA‐S1 type strains. The induction of toxic oxygen radicals byH. pylori‐activatedneutrophils gave higher median values for thecagA‐positive strains than for thecagA‐negative strains. For the monocyte chemiluminescence response,cagA‐negative strains gave higher median values compared tocagA‐positive strains. We conclude that upregulation of the neutrophil and monocyte adherence molecule CDllb induced byH. pylorisonicates is not associated with the presence ofcagA,picB or mosaic pattern ofvacA, and thatcagA,picB‐negative strains andvacA‐S2 strains retain their inflammatory capacity.
Blood levels of CD11bH+ memory T lymphocytes are selectively upregulated in patients with active rheumatoid arthritis
Authors: Nielsen Henrik , Petersen Ane Ahm , SkjøDt Henrik , HøRslev‐Petersen Kim , Bendtzen Klaus
Published: August 2009

The adhesion molecules CD11b (a β2‐integrin component) and CD54 (ICAM‐1) on blood leukocytes were studied by flow cytometry in patients with rheumatoid arthritis (RA). The fractions of CD4+ cells co‐expressing CD11b were elevated in 16 patients with active RA compared with those in 16 RA patients who improved during therapy and 8 healthy controls: 0.8±0.12% (mean±M) versus 0.3±0.06% (p<0.002) and 0.3±0.06% (p<0.005), respectively. Increased levels of CD1 1b+CD45R0+ cells were observed in patients with active RA compared to those with improved RA and controls: 12.6±3.9% versus 4.8±2.7% (p<0.002) and 6.1±1.2% (p<0.003), respectively. Disease activity, determined by C‐reactive protein, correlated with the numbers of CD11b+CD45R0+ cells: r=0.62 (p<0.001). Seven patients were followed during induction of remission with methotrexate and glucocorticoids. The numbers of CD11b+CD4+ and CD11b+CD45R0+ cells fell significantly after clinical improvement. The levels of CD11b+CD14+ cells (monocytes) did not differ between the groups. The number of CD11b+CD15+ cells (neutrophils) was elevated in patients with RA irrespective of disease activity. The levels of CD54+ cells were not different between the RA and control groups. We conclude that the increased numbers of CD11b+ memory T cells may arise from exposure to stimuli outside the synovial compartment.
Endoproteinase‐protein inhibitor interactions
Authors: BODE WOLFRAM , FERNANDEZ‐CATALAN CARLOS , NAGASE HIDEAKI , MASKOS KLAUS
Published: August 2009

Nature uses protein inhibitors as important tools to regulate the proteolytic activity of their target proteinases. Most of these inhibitors for which 3D structures are available are directed towards serine proteinases, interacting with their active‐sites in a substrate‐like “canonical” manner via an exposed reactive‐site loop of conserved conformation. More recently, some non‐canonically binding serine proteinase inhibitors, two cysteine proteinase inhibitors, and three zinc endopeptidase inhibitors have been characterized in the free and complexed state, displaying novel mechanisms of inhibition with their target proteinases. These different interaction modes are briefly discussed, with particular emphasis on the interaction between matrix metalloproteinases (MMPs) and their endogenous tissue inhibitors of metalloproteinases (TIMPs).