bluuebunny/crossref_metadata_2025 huggingface dataset:
Legionella species of different human prevalence induce different rates of apoptosis in human monocytic cells
Authors: Walz J. M. , Gerhardt H. , Faigle M. , Wolburg H. , Neumeister B.
Published: October 2003

Legionella species of different human prevalence were examined with respect to induction of apoptosis in the human monocytic cell line Mono Mac 6 (MM6). L. pneumophila serogroup 1 (Pontiac), L. pneumophila serogroup 1 (Philadelphia‐1), L. longbeachae serogroup 1, L. gormanii, L. micdadei and L. steigerwaltii were used to infect MM6 cells. Subsequent induction of apoptosis was investigated by enzyme‐linked immunosorbent assay (ELISA), gel electrophoresis of cellular DNA extracts, and staining of cells with the DNA dye 4′, 6‐diamidino‐2‐phenylindole (DAPI). Additionally, the concomitant occurrence of infection and apoptosis was demonstrated by a combination of immunohistochemistry with nuclear DAPI counterstaining. Induction of apoptosis in MM6 cells by a given species of the genus Legionella correlates with their human prevalence rather than with their ability to multiply within this human monocytic cell line. Furthermore, we found that initiation of apoptosis of Mono Mac 6 cells was dependent on direct adherence of the pathogenic bacteria to the host cell and was triggered by extracellular bacteria.
Reduced ammonium chloride haemolysis time enhances the number of isolated functional rabbit polymorphonuclear neutrophils
Authors: Kouoh Ferdinand , Levert Helene , Gressier Bernard , Luyckx Michel , Brunet Claude , Dine Thierry , Ballester Louis , Cazin Micheline , Cazin Jean Claude
Published: October 2003

In the present study we compared seven different methods for isolating rabbit polymorphonuclear neutrophils (PMNs) with a view to assessing viability, lymphocyte contamination and isolation yield. The two methods offering the best isolation yield and functional PMNs were retained. Leukocyte‐containing plasma fraction was obtained after erythrocyte sedimentation with dextran. First, PMNs were isolated from this fraction, using hypotonic ammonium chloride haemolysis followed by Histopaque® density gradient centrifugation (Method‐A). Second, PMNs were obtained from leukocyte‐containing plasma after centrifugation on two Percoll® layers (Method‐B). These processes resulted in a high cellular yield: 2.66±106±0.22 PMNs per ml of blood (Method‐A) and 1.87±106±0.37 PMNs per ml of blood (Method‐B). In both cases the PMNs isolated were of high purity and viability. In comparison, when using the standard techniques for rabbit ‐ consisting of ammonium chloride haemolysis taking at least four times as long ‐ fewer PMNs were isolated. The PMNs isolated by Method‐A and ‐B were able to generate a high amount of reactive oxygen species (ROS) after stimulation with phorbol 12‐myristate 13‐acetate (PMA). These methods to separate PMNs are recommended for in vitro studies.
Prenatal malformed lumbar vertebral corpora in trisomies 21, 18 and 13, evaluated radiographically and histologically
Authors: Nolting Dorrit , Hansen Birgit Fischer , Keeling Jean W. , Reintoft Ingermarie , KjæR Inger
Published: October 2003

The aim of the present study was to compare, both radiographically and histologically, malformed vertebral lumbar corpora in trisomies 21, 18 and 13 with earlier reported normal corporal development in the axial lumbar region. Axial skeletons of human fetuses (GA 15–22 wk) derived from therapeutically induced abortion were investigated in connection with requested autopsy. The number of lumbar vertebral corpora examined for each genotype was as follows: 20 from trisomy 21, 10 from trisomy 18, and 10 from trisomy 13. After radiography in frontal, lateral and axial projections, the individual vertebral corpora were decalcified and horizontally embedded in paraffin. The blocks were serially sectioned and stained with toluidine blue and alcian blue/van Gieson. The radiographic characteristics of the vertebral corpora varied from an almost normal appearance of the corporal bone to complete clefting of the bony corpora. Histological examination showed accumulations of cartilage centrally, in some cases associated with amorphous material. Pronounced metachromatic differences were observed in the cartilaginous ground substance. The study showed identical phenotypic characteristics in the corpora from trisomy 21, trisomy 18, and trisomy 13. It is characteristic of all three genotypes that there are central anomalies, corresponding to the location of the notochord in normal corpora, and marked regional differences in metachromasia in the ground substance of the cartilage.
Sumatriptan increases the proliferation of peripheral blood mononuclear cells from HIV‐infected individuals and healthy blood donors in vitro
Authors: Afzelius Pia , Nielsen Jens Ole
Published: October 2003

HIV infection is characterized by the loss of CD4+ T cells as well as the loss of T‐cell function, leading to severe immunodeficiency. The proliferative capacity of T cells measured in vitro as responses to antigens and mitogens is severely reduced during HIV infection. An increased level of the intracellular second messenger adenosine 3′,5′‐cyclic monophosphate (cAMP) has been shown to cause impaired proliferative capacity of peripheral blood mononuclear cells (PBMC) from HIV‐infected individuals in vitro. Sumatriptan, a 5HT1d receptor agonist, inhibits the activity of adenylyl cyclases, the enzymes responsible for regulation of the intracellular levels of cAMP. In a preliminary study sumatriptan increased the proliferative responses of PBMC to a polyclonal activator in vitro in 9 of 10 HIV‐seropositive individuals (p=0.007), and in 7 of 9 healthy blood donors (p=0.05). This was probably due to a decrease in the intracellular level of cyclic AMP.
Modes of adherence of Helicobacter pylori to gastric surface epithelium in gastroduodenal disease: A possible sequence of events leading to internalisation
Authors: Papadogiannakis Nikos , Willen Roger , Carlen Birgitta , SjÖStedt Svante , WadstrÖM Torkel , Gad Adel
Published: October 2003

We have investigated various modes of adherence of Helicobacter pylori to the human gastric epithelium, using transmission electron microscopy, in biopsies from nine patients with peptic ulcer disease and from four patients with chronic active gastritis. H. pylori was demonstrated in abundance in all cases within the surface mucous layer. In all ulcer‐ and in one out of four gastritis patients H. pylori was shown in close proximity to the gastric epithelium, with concurrent alterations in the configuration of microvilli and the apical cytoplasmic region of gastric cells. Previously described modes of H. pylori adherence were confirmed, such as loose attachment with fibrillar‐like strands, firm attachment with pedestal formation, invasion in the intercellular spaces, and invagination with “cup” formation. Moreover, in many cases a fusion between the bacterial outer layer and gastric cell membranes was evident. In four cases (31%; three with active and one with past ulcer disease) viable H. pylori was found in the cytoplasm of gastric mucous cells. Our results support the hypothesis that the different modes of adherence of H. pylori represent a stepwise, possibly sequential, process which in a significant number of cases leads to internalisation of the organism. The invariable occurrence of adhesion and more frequent internalisation of H. pylori in ulcer patients may suggest a link with the pathogenesis of peptic ulcer disease.
Fibronectin‐binding proteins secreted by Mycobacterium avium
Authors: Kitaura HIDEKI , Ohara NAOYA , Naito MARIKO , Kobayashi KAZUHIDE , Yamada TAKESHI
Published: October 2003

Mycobacterium avium is an intracellular pathogen and a major opportunistic infectious agent observed in patients with acquired immune deficiency syndrome (AIDS). Fibronectin is an extracellular matrix protein and is a virulence factor for several extracellular pathogenic bacteria binding to mucosal surfaces. We investigated the fibronectin (FN)‐binding proteins in the culture filtrate of M. avium by two‐dimensional electrophoresis (2DE). Proteins in Sauton medium of M. avium after 3 weeks were separated by 2DE. The proteins were blotted onto polyvinylidene difluoride membrane and incubated with FN. FN‐binding proteins were detected by Western blotting using anti‐FN antibody. FN bound to five spots (33 kDa, 32 kDa, 31 kDa, 30 kDa and 25 kDa). N‐terminal amino acids of these were determined. The 33 kDa spot corresponded to antigen 85 (Ag 85) C. The 32 and 31 kDa spots were either Ag 85 A or Ag 85 B. The 30 kDa spot corresponded to Ag 85 B of M. avium. The 25 kDa spot corresponded to MPA51 (M. avium MPB51). Thus, FN bound exclusively to the Ag 85 complex and MPA51.
Antimicrobial resistance of Shigella sonnei in Korea during the last two decadesNote
Authors: Lee Je Chul , Oh Jae Young , Kim Ki Sung , Jeong Yong Wook , Cho Jae We , Park Jong Chun , Seol Sung Yong , Cho Dong Taek
Published: October 2003

Eighty‐eight strains of Shigella sonnei isolated in Korea during the period 1980 to 1999 were tested for susceptibility to 13 antimicrobial agents. S. sonnei isolates demonstrated high frequencies of resistance to sulfamethoxazole (97.7%), tetracycline (96.6%), and trimethoprim (95.5%). S. sonnei isolates from the 1990s were more resistant to nalidixic acid than isolates from the 1980s (100 vs 7.7%), while isolates from the 1990s were more susceptible to chloramphenicol than isolates from the 1980s (0 vs 100%). Ampicillin‐resistant S. sonnei isolates produced the TEM‐1 ß‐lactamase with a pI of 5.4. The TEM‐1 gene was located on conjugally transferable plasmids in the majority of isolates. S. sonnei isolates were all susceptible to cefotaxime, cefoxitin, ceftazidime, ciprofloxacin, and norfloxacin. These results indicate that cephalosporins and quinolones may be alternative antibiotics for the treatment of S. sonnei infections in Korea.
Epidemiology of breast cancer: an environmental disease?Note
Authors: Sasco Annie J.
Published: October 2003

Breast cancer is the leading cancer site in women, both in the developed and the developing world. Incidence rates are increasing in many countries, although, in some, mortality may be stable or slightly decreasing. Geographical differences exist, with high rates of disease in North America, North Europe and Oceania, intermediate rates in South and Central America as well as South and East Europe, and low rates in Africa and Asia. Most of the literature reports that genetic inherited factors account for less than 5% of cases, although some authors advance higher figures, up to about 10%. Risk factors for breast cancer are related to the reproductive life of women: early menarche, nulliparity or late age at first birth, late menopause, diet and physical exercise, as well as hormonal factors, be they endogenous (high levels of free or not bound to SHBG estrogens) or exogenous (long‐term use of oral contraceptives or menopausal hormone replacement). The present review does not aim to be exhaustive and fully comprehensive, or to present in detail domains currently well known and accepted by all. On the contrary, it modestly wishes to highlight potentially controversial conditions which could in the future be recognized as new risk factors.
Arbitrarily primed PCR and sequencing of 16S rDNA for epidemiological typing and species identification of Burkholderia cepacia isolates from Swedish patients with cystic fibrosis reveal genetic heterogeneityNote
Authors: Karpati Ferenc , Giedraitis Vilmantas , Thore Magnus , Lindman Runa , Monstein Hans‐Jürg , Hjelte Lena , Jonasson Jon
Published: October 2003

To investigate whether arbitrarily primed (AP)‐PCR and/or 16S rDNA sequencing could be used as rapid methods for epidemiological typing and species identification of clinical Burkholderia isolates from patients with cystic fibrosis (CF), a total of 39 clinical B. cepacia isolates, including 33 isolates from 14 CF patients, were fingerprinted. ERIC‐2 primer was used for AP‐PCR. The AP‐PCR clustering analysis resulted in 14 different clusters at a 70% similarity level. The AP‐PRC patterns were individual despite considerable similarities. To sequence rDNA, a broad‐range PCR was applied. The PCR product included four variable loops (V8, V3, V4 and V9) of the 16S ribosomal small subunit RNA gene. The multiple sequence alignment produced 12 different patterns, 5 of them including more than one isolate. Heterogeneity of the bases in the V3 region, indicating the simultaneous presence of at least two different types of 16S rRNA genes in the same cell, was revealed in 10 isolates. Most of the CF patients were adults who had advanced disease at follow‐up. Both the sequencing and the AP‐PCR patterns revealed genetic heterogeneity of isolates between patients. According to the results obtained, AP‐PCR could advantageously be used for epidemiological typing of Burkholderia, whereas partial species identification could effectively be obtained by sequencing of the V3 region of the 16S RNA gene.
Macrophage and lymphocyte chimerism in bronchoalveolar lavage cells from human lung allograft recipients
Authors: Wiebe Britt Marie , Mortensen Svend Aage , Petterson Gösta , Svendsen Ulrik Gerner , Andersen Claus Bøgelund
Published: October 2003

Background. Chimerism is suggested to predict a more favourable prognosis in solid organ transplantation. Material and method. Forty‐eight bronchoalveolar lavages from 10 patients (5 females and 5 males) who had received sex‐mismatched donor lungs were monitored for varying periods of time, of up to 2 years, at regular intervals (median 3.0 (0.5–24) months). To investigate the chimerism in macrophages and lymphocytes in bronchoalveolar lavage cells a cloned 2.12 kilobase large biotinylated Y‐chromosome‐specific DNA‐probe was used for in situ hybridization. Results. Donor macrophages disappeared in seven patients within the first 6 months after surgery (median 3.0 (1–6) months). But 15% donor macrophages could be detected in one patient 1 year and 10% in 2 patients two years after surgery. Donor lymphocytes disappeared in all patients within 3 months (median 1 (0.5–3) months). There was no correlation between periods or severity of acute rejection and percentage of donor macrophages and donor lymphocytes in bronchoalveolar lavage. None of the patients developed obliterative bronchiolitis. Conclusion. Macrophage chimerism in lung may exist for several years. Whilst our results do not elucidate the role of local macrophage chimerism, they do not currently support the view that chimerism prevents rejection.
Genotypic detection of enterotoxigenic Escherichia coli colonisation factors
Authors: Grewal Harleen M. S. , Sommerfelt Halvor
Published: October 2003

We have developed a nonradioactive colony hybridisation assay for the detection of enterotoxigenic Escherichia coli (ETEC) that harbor the structural genes for CFA/I, CS1, CS2, CS4, CS17, or PCFO166. Thus, a polynucleotide probe derived from the colonisation factor antigen I (CFA/I) operon hybridised under very low stringency conditions to total DNA from CFA/I‐producing (CFA/I), coli‐surface antigen 1 and 3 (CS1 CS3‐), CS2 CS3‐, CS4 CS6‐, CS17‐, and putative colonisation factor O166 (PCFO166)–producing enterotoxigenic Escherichia coli (ETEC). The probe did not hybridise to DNA from CS3, CFA/III CS6, CS5 CS6, CS6, CS7, or PCFO159 ETEC. Visual registration of colour intensity could be used to differentiate between CFA/I, CS4 and PCFO166‐positive strains on the one hand and strains with the genetic potential to express CS1, CS2, or CS17 on the other. As a confirmatory test, restriction fragment patterns obtained from Sau3AI‐digested ETEC plasmid DNA could be used to distinguish between CFA/I, CS1, CS4, CS17, and PCFO166 ETEC in nonradioactive Southern blot hybridisation. The simultaneous genotypic detection of several ETEC colonisation factors will prove useful in vaccine‐oriented studies of ETEC disease.
Allele 2 of interleukin‐1 receptor antagonist gene increases the risk of thyroid peroxidase antibodies in subacute thyroiditis
Authors: Luotola K. , Mantula P. , Salmi J. , Haapala A.‐M. , Laippala P. , Hurme M.
Published: October 2003

Subacute granulomatous thyroiditis (SAT) is a self‐limiting systemic inflammatory disorder with possible transient expression of thyroid antibodies. Persistent hypothyroidism is uncommon. The interleukin‐1 receptor antagonist IL‐1ra is an inhibitor of IL‐1 activity and allele 2 of the IL‐1ra gene is associated with inflammatory diseases and IL‐1ra production. Forty‐eight subjects with SAT were investigated. Polymorphisms of IL‐1ra, IL‐1β‐511 and TNFα genes were studied with respect to thyroid peroxidase antibodies (TPOab), thyroglobulin antibodies, C‐reactive protein (CRP) and erythrocyte sedimentation rate (ESR). Serum IL‐1ra levels were measured. An increased allelic frequency (43% vs 22%, p=0.039) and carriage rate (79% vs 41%, p=0.018) for IL‐1ra allele 2 were observed with expression of TPOab in 14 (29.2%) subjects compared with those with negative findings. The carriage rate for both IL‐1ra allele 2 and IL‐1β‐511 allele 2 was also increased with TPOab expression (71% vs 27%, p=0.004, respectively). No difference in allele frequency or carriage rate was found compared with healthy controls. Serum IL‐1ra levels correlated with S‐CRP (r=0.41, p=0.004) and ESR (r=0.34, p=0.016), but the association with genes or thyroid antibodies was statistically insignificant. S‐CRP levels and ESR were lower and negatively correlated with expression of TPOab (r=−0.27, p=0.046 and r=−0.32, p=0.017). This study describes the multiplicity of the mechanisms responsible for the severity of the acute‐phase response during the course of SAT. IL‐1ra may have a significant anti‐inflammatory role in SAT. Presence of IL‐1ra allele 2 increases the risk of developing TPOab.
Investigation of an outbreak caused by methicillin‐resistant Staphylococcus aureus in a cardiovascular surgery unit by ribotyping, randomly amplifed polymorphic DNA and pulsed‐field gel electrophoresis
Authors: Fung Chang‐Phone , Ho Mao‐Wang , Wang Fu‐Der , Tsai Kevin , Liu Chun‐Eng , Liu Cheng‐Yi , Siu L. K.
Published: October 2003

An outbreak caused by rapid spread of methicillin‐resistant Staphylococcus aureus (MRSA) in an intensive care unit for cardiovascular surgery was investigated by phenotypic and genotypic methods. Fourteen isolates were collected during a 2‐month period from clinical and environmental specimens in the unit recently re‐opened after reconstruction. The isolates were tested for antibiotic susceptibility patterns and genotyped by automated ribotyping, randomly amplified polymorphic DNA‐PCR (RAPD) analysis and pulsed‐field gel electrophoresis (PFGE). Automated ribotyping applying EcoRI digestion proved to be of no value in separating the isolates. In contrast, PFGE grouped the isolates into four clusters different from the reference strain. These results fully correlated with the antibiograms. Twelve of the isolates were grouped into two clonally related clusters. RAPD analyses grouped the isolates into five clusters. Except for two isolates of one patient, which had different RAPD patterns, PFGE and RAPD analyses presented very similar results. The results verified the usefulness of PFGE in studies of MRSA epidemics. A combination of these two methods reduces the time to identification of an outbreak and increases the accuracy in detection of intraspecies differences.
Invitro Candidacidal effect of polymorphonuclear neutrophils of Behçet's patients with or without ocular involvementNote
Authors: TEKELİ ALPER , TEKELİ OYA , ÖZDEMİR ÖZDEN
Published: October 2003

In this study our objective was to evaluate the in vitro candidacidal effect of polymorphonuclear neutrophils of Behçet's patients with or without ocular involvement (inactive phase). Fifteen patients with ocular involvement and 15 patients without ocular involvement were studied. Candidacidal assay was performed to assess the in vitro killing of Candida species by the polymorphonuclear neutrophils of both the study and control groups. In brief, peripheral venous polymorphonuclear neutrophils of healthy volunteers and patients with Behçet's disease were obtained by density gradient centrifugation. These cells were incubated with Candida spp. (Candida albicans, Candida tropicalis, Candida glabrata) for 3 h. Samples were seeded onto yeast extract peptone dextrose agar, incubated at 25 °C in a 5% humidified incubator for another 48 h, and formed colonies were counted. There was no significant difference between the candidacidal activity of polymorphonuclear neutrophils of healthy volunteers and of Behçet's patients with or without ocular involvement. Although various immunological abnormalities are reported in Behçet's disease, in vitro killing of Candida spp., which is one of the parameters for the phagocytic function of polymorphonuclear neutrophils, was not affected.
Pseudoangiomatous hyperplasia of mammary stroma: A case of pure type after removal of fibroadenomaNote
Authors: FUKUNAGA MASAHARU
Published: October 2003

A case of pure pseudoangiomatous hyperplasia of the mammary stroma after removal of a fibroadenoma is described. The lesion, which was found in the right breast of a 40‐year‐old woman, was a well‐circumscribed non‐encapsulated, rubbery, lobulated mass measuring 40×40×35 mm. Histologically, it consisted predominantly of a proliferation of spindle cells with interanastomosing vascular‐like arrangements in the interlobular or interductal stroma. Neither cytological atypia nor mitotic figures were observed. The stroma contained abundant collagen with focal hyalinization. Focally, epithelial elements showed mild ductal hyperplasia. There was no fibroadenoma. Immunohistochemically, the spindle cells were positive for vimentin, CD34, α‐smooth muscle actin, muscle actin, CD34, calponin, and progesterone receptors. Ultrastructurally, many spindle cells had thin elongated cytoplasmic processes, which enveloped pseudovascular spaces containing a few collagenous fibrils, indicating fibroblastic cells. This lesion should be distinguished from other mammary fibrous and vascular lesions with which it may be confused.
Insulin‐like growth factor‐II in endocrine pancreatic tumoursNote
Authors: HÖÖG ANDERS , KJELLMAN MAGNUS , NORDQVIST ANN‐CHRISTIN SANDBERG , HÖÖG CHARLOTTE M. , JUHLIN CLAES , FALKMER STURE , SCHALLING MARTIN , GRIMELIUS LARS
Published: October 2003

In earlier studies a high‐molecular‐weight (HMW) insulin‐like growth factor‐II (IGF‐II) peptide was identified in adult human pancreas and localized to the insulin‐producing B‐cells. This peptide has now been investigated in neoplastic insulin cells. Forty endocrine pancreatic tumours and 17 pancreatic adenocarcinomas of ductal type were included in the study. All cases were investigated with immunohistochemical techniques using antibodies to IGF‐II, insulin, pro‐insulin, glucagon, somatostatin, pancreatic polypeptide, gastrin and vasoactive intestinal peptide (VIP). Frozen tissue from nine tumours and two normal pancreatic glands was extracted, gel separated, and quantified using radioimmunoassay. The tumours were also investigated by in situ hybridization. IGF‐II‐immunoreactive cells were found in nearly all the 18 insulin‐producing tumours (16/18), in a minority of the other endocrine tumours, but not in pancreatic adenocarcinomas. All extracts from the endocrine tumours showed varying amounts of IGF‐II and had different molecular‐weight forms. The immunohistochemical and radioimmunoassay findings are both based on immunological binding and were further confirmed by Northern blot and in situ hybridization. These results show that IGF‐II is expressed in insulin‐producing tumours as well as in pancreatic tumours producing other peptides, in contrast to normal pancreatic islets where IGF‐II is found exclusively in insulin‐producing cells.
Absence of group II phospholipase A2, a Paneth cell marker, from the epididymisNote
Authors: Nevalainen Timo J. , Shah Varsha I. , De Peralta‐Venturina Mariza , Amin Mahul B.
Published: October 2003

Paneth cell‐like metaplasia has been reported in the epithelium of the epididymis and prostatic adenocarcinomas. We studied the expression of group II phospholipase A2 (PLA2), a marker of Paneth cell differentiation, in six orchiectomy specimens with Paneth cell‐like metaplasia. Both immunohistochemistry for group II PLA2 protein and in situ hybridization for the mRNA of group II PLA2 gave negative results in all six cases but positive reaction for lysozyme. The results show that the cells of the Paneth cell‐like metaplasia are not true Paneth cells.
A test for cleaning and disinfection processes in a washer‐disinfectorNote
Authors: Ransjö Ulrika , Engström Lars , Håkansson Paul , Ledel Torbjörn , Lindgren Lisa , Lindqvist Anna‐Lisa , Marcusson Eva , Rudbäck Klas
Published: October 2003

Disinfection processes such as heat, aldehydes or alcohols kill vegetative microorganisms but do not necessarily remove other organic contamination. Organic residues impair the result of low‐temperature sterilisation processes. Heat‐stable organic residues may give rise to clinical symptoms in the patient. Standards are available in Britain and in Sweden for the examination of cleaning processes in washer‐disinfectors. The test substances are artificial soil or blood. These standards are based on visual inspection of instruments or equipment. They cannot be used for examination of tubular instruments, nor can they be quantified. For validation of cleaning procedures a simple quantifiable method, which can be performed in an infection control laboratory is needed. We have used suspensions in horse blood of Enterococcus faecalis bacteria and Bacillus subtilis spores to test disinfection and cleaning in a washer‐disinfector. Instruments used for laparoscopic surgery were contaminated with a blood–bacteria suspension containing 107 organisms/ml and then dried and processed in a washer‐disinfector using a regular process. Remaining microbial contamination was cultured quantitatively. Nineteen objects were investigated in 10 experiments each. Cleaning, measured as log reduction >5–6 of B. subtilis, was achieved on surfaces that were adequately in contact with the water flow in the machine. Disinfection (and cleaning) measured as log reduction >5–6 of E. faecalis was successful at all points examined. The test method is simple and quantifiable, and can be used to evaluate and to improve cleaning and disinfection processes.
Antibiotic susceptibility of blood culture isolates of EnterobacteriaceaeNote
Authors: DIGRANES ASBJØRN , SJURSEN HAAKON , MELBY KJETIL K. , SOLBERG CLAUS OLA
Published: October 2003

From May to November 1997 each of six major hospitals throughout Norway collected 72 to 104 consecutive blood culture isolates of Enterobacteriaceae, altogether 563 isolates. Escherichia coli was the predominating organism (69%), followed by Klebsiella spp. (15%), Enterobacter spp. (6%), and Proteus mirabilis (4%). The susceptibility of the isolates to ampicillin, cefuroxime, ceftazidime, imipenem, tobramycin, and ciprofloxacin was determined by the E‐test. 37% and 7% of the isolates were resistant to ampicillin and cefuroxime, respectively, and 1% were resistant to ceftazidime and tobramycin. Only one isolate of P. mirabilis was imipenem resistant. All isolates were susceptible to ciprofloxacin. The prevalence of ampicillin‐resistant isolates at each hospital varied from 21 to 45%, and of cefuroxime‐resistant isolates from 3 to 9%. The results were compared with those of a similar study performed in 1991–1992. No significant changes in the susceptibility to the various agents could be demonstrated. The high frequency of isolates resistant to ampicillin has clearly limited the usefulness of this agent in the treatment of septicemia and other serious infections caused by Enterobacteriaceae.
Production and partial characterization of mouse monoclonal antibodies recognizing common cytokine receptor gamma chain (γc) of human, mouse and primate originNote
Authors: LUNDIN KAROLINA , LEPPIMÄKI PETRA , OETKEN CHRISTINA , NORDSTRÖM TOMMY , LINDQVIST CHRISTER
Published: October 2003

Monoclonal antibodies specific for the common cytokine receptor gamma chain, γc, were produced using traditional hybridoma technology. Fusion of P3X63‐Ag8.653 myeloma cells with splenocytes from Balb/c mice immunized with Spodoptera frugiperda insect cells infected with the recombinant baculovirus VL1392‐hIL‐2Rγ resulted in several hybridoma cell clones producing monoclonal γc‐specific antibodies. Four of these antibody‐producing clones, IIIC3, IIIE8, IG3 and IF10C5, were further characterized by immunoblotting, flow cytometry and ELISA. Data are presented demonstrating that the generated monoclonal antibodies can identify the extracellular domain of the common cytokine receptor γ chain of human and mouse origin, and two of the antibodies recognize γc of primate origin as well.
Apoptotic cell death induced by Acinetobacter baumannii in epithelial cells through caspase‐3 activationNote
Authors: LEE J. C. , OH J. Y. , KIM K. S. , JEONG Y. W. , PARK J. C. , CHO J. W.
Published: October 2003

Epithelial cell death induced by Acinetobacter baumannii infection was investigated using in vitro assays. Eight hours after live A. baumannii infection, HeLa cells exhibited detachment from the dish, rounding morphologies, high proportions of trypan blue‐positive cells and extensive DNA breakdown with faint apoptotic banding, which is indicative of cells undergoing apoptosis. The enzymatic activity of caspase‐3 was increased in cells as early as 2 h after infection. In addition, apoptosis of HeLa cells was induced by treatment with bacterial culture filtrates but not with formalin‐killed bacteria. These results indicate that A. baumannii infection triggers apoptosis in HeLa cells through caspase‐3 activation.
Factors affecting the cytology outcome of Pap smears – a brief approach to internal quality control in private cytopathology laboratory practiceNote
Authors: LINKO SOLVEIG , TASKINEN EERO , SARNA SEPPO , KÄRKKÄINEN PÄIVI
Published: October 2003

We evaluated the effect of intrauterine device (IUD), patient age and hormone replacement therapy (HRT) on cytology outcome in Pap smears together with the important IQC procedures: 1) manual double‐screening by cytotechologists, and 2) retrospective senior pathologist review during 1996–1999. The results from primary double‐screening (119 of 87,409 Pap smears) showed an excellent interobserver correlation. The estimation of hormonal effects showed higher incidence of disagreements (p=0.013) in patients <47 yr. Some individual trends were found in the assessments of both cellular atypia (p=0.012) and Papanicolaou classification (p=0.018). The IUD had no influence on the accuracy when the degree of inflammatory reaction was evaluated (p>0.050), but showed an adverse effect on the estimation of cellular atypia (p=0.001). HRT distinctly equalized the entire sample material, since fewer disagreements were found in the age groups <47 yr and >47 yr when estimating the hormonal effects (p=0.013), inflammatory reaction (p=0.044) or cellular atypia (p=0.006) compared to those without HRT. The continuous cytopathologist supervision had a positive impact on the accuracy of hormonal effect estimation during the 4 years. The senior cytopathologists' reviews (354 of 87,409 Pap smears) showed mutually good interobserver correlation, and diagnostic conclusions of the same specimens differed only slightly between the cytopathologists. We found these state‐of‐the‐art cytopathological IQC procedures to be effective and fit‐for‐purpose when evaluating hormonal effects, inflammatory reaction and cellular atypia.
Poorly differentiated endometrial sarcoma with clear cell featuresNote
Authors: FUKUNAGA MASAHARU
Published: October 2003

A case of poorly differentiated endometrial sarcoma with clear cell features in a 46‐year‐old female is presented. A 9×8×6 cm, transmural, white solid tumor in the uterine fundic wall was characterized histologically by infiltrative growth and a solid arrangement of tumor cells with clear cytoplasm. The tumor showed vascular invasion, moderate to severe cellular atypia, mitotic activity of 20 per 10 high‐power fields and necrosis. It was immunohistochemically positive for vimentin and negative for CAM5.2, epithelial membrane antigen, desmin, alpha‐smooth muscle actin, muscle actin, estrogen and progesterone receptors. Electron microscopy showed that the clear cytoplasm was attributed to the presence of numerous dilated mitochondria and lipid droplets. Flow cytometry showed a diploid phenotype. Pathologists should be aware of the existence of this type of endometrial sarcoma with clear cells and should not confuse it with other neoplasma showing clear cell features.
Angioimmunoblastic T‐cell lymphoma with hyperplastic germinal centers: a clinicopathological and immunohistochemical study of 10 casesNote
Authors: KOJIMA MASARU , NAKAMURA SHIGEO , ITOH HIDEAKI , MOTOORI TADASHI , SUGIHARA SHIRO , SHINKAI HIROKO , MASAWA NOBUHIDE
Published: October 2003

An absence of germinal centers is one of the histological characteristics of angioimmunoblastic T‐cell lymphoma (AITL). We report here 10 unusual cases of AITL with hyperplastic germinal centers. The clinical presentation of each patient was characterized by generalized lymphadenopathy, constitutional symptoms and polyclonal hypergammaglobulinemia. The initial biopsy findings of each patient were similar and were characterized by hyperplastic germinal centers with ill‐defined borders and a proliferation of high endothelial venules (HEV). In the paracortical area there was a mixed infiltrate including irregularly shaped clusters or small nests of clear cells in all cases. Moreover, the clear cells invaded the lymphoid follicles, resulting in expansion of the germinal centers, except for one case. Immunohistochemistry revealed that the tumor cells, including clear cells, were CD4‐expressing T cells. Some of the atypical lymphocytes were also Bcl‐6‐positive. A majority of the follicular dendritic cell networks showed a normal/reactive or an expanded/disrupted pattern in all cases. Moreover, three lesions possessed a few large irregularly shaped proliferations of follicular dendritic cells around the HEV. Four cases progressed to AITL within a few years. The present 10 cases probably represent an early stage of AITL preceding follicular dendritic cell hyperplasia. Detection of clear cells, Bcl‐6‐positive atypical T lymphocytes, and foci of irregularly shaped proliferation of follicular dendritic cells appears to be critical for early diagnosis and treatment of AITL with hyperplastic follicles.
Analysis of Borrelia burgdorferi sensu lato isolated from cerebrospinal fluidNote
Authors: RUŽIĆ‐SABLJIĆ EVA , LOTRIČ‐FURLAN STANKA , MARASPIN VERA , CIMPERMAN JOŽE , PLETERSKI‐RIGLER DUŠICA , STRLE FRANC
Published: October 2003

Involvement of the nervous system in Lyme borreliosis may occur with or without erythema migrans and it may present with a variety of neurological symptoms. In this study we analysed phenotypic and genotypic characteristics of 40 Borrelia strains isolated from cerebrospinal fluid (CSF) of 38 Slovenian patients with different clinical manifestations of Lyme borreliosis. In seven of the patients, Borreliae were also isolated from skin lesions. Species identification and plasmid profiles were determined by pulsed‐field gel electrophoresis and protein profiles by SDS‐PAGE. MluI digestion profiles of Borrelia burgdorferi sensu lato DNA showed that 25 (62.5%) isolates were B. garinii, 14 (35%) B. afzelii, and one (2.5%) B. burgdorferi sensu stricto. All strains, except one, possessed a large plasmid and a varying number of smaller plasmids. Three (7.5%) isolates exhibited an unusual plasmid profile, with a large plasmid dimer or three copies of the large plasmid. In protein analyses, all strains expressed OspA protein. OspB was present significantly more often in B. afzelii than B. garinii strains (p=0.0000), while OspC was more often present in B. garinii than B. afzelii strains (p=0.0052). In the seven patients with Borreliae isolated also from the skin, the CSF and skin isolates were identical, either B. garinii (six patients) or B. afzelii (one patient). Species and plasmid heterogeneity as well as antigen diversity could play a role in the pathogenesis of the infection. When combined with our own earlier data, the results suggest species‐related organotropism.
Malignant neoplasms: discordance between clinical diagnoses and autopsy findings in 3,118 cases
Authors: AVGERINOS DIMITRIOS V. , BJÖRNSSON JOHANNES
Published: October 2003

Background. During the past few decades, hospital autopsy rates have steadily declined throughout the Western world. This decline is mainly attributed to the introduction of advanced diagnostic techniques. Despite technological developments, discrepancy rates between clinical diagnoses and autopsy findings remain high. Few studies have addressed discrepancy rates exclusively with regard to malignant neoplasms. In the present study, we reviewed the records of 3,118 autopsies performed at Mayo Clinic during a 6‐year period (1994–1999) and identified clinically undiagnosed malignancies found at autopsy and clinically diagnosed cancers not confirmed at postmortem examination. Materials and Methods. Autopsy protocols, provisional and final anatomic diagnoses, and data from the Mayo Autopsy Pathology Quality Assurance program were reviewed in an attempt to identify discrepancies between clinical diagnoses and autopsy findings regarding malignant neoplasms. Results. In 3,118 autopsies performed at Mayo Clinic between 1994 and 1999, a malignant tumor was identified in 768 cases (25%). In 128 of 3,118 cases (4.1%), the malignancy was not diagnosed clinically. In 14 of 3,118 cases (0.45%), autopsy failed to confirm a clinically diagnosed cancer. A review of the literature is presented. Conclusions. Autopsy remains an effective tool for the confirmation and refutation of clinical diagnostic findings regarding malignant neoplasms.
An association between chronic infection with Chlamydia pneumoniae and lung cancer. A prospective 2‐year studyNote
Authors: KOYI HIRSH , BRANDÉN EVA , GNARPE JUDY , GNARPE HÅKAN , STEEN BERTIL
Published: October 2003

This study assesses a possible relationship between chronic Chlamydia pneumoniae (Cpn) infection and lung cancer (LC). A total of 210 consecutive patients (136 M, 74 F) were diagnosed with LC during a 2‐year period. Blood was obtained from 128 M and 70 F patients for Cpn serology. Repeat blood specimens were taken after 3 months. Throat specimens for Cpn DNA analysis by PCR were taken from 110/136 M and 63/74 F. Seventy‐four cytobrush specimens were taken and also analyzed by polymerase chain reaction (PCR). Fifty (29 M, 21 F) bronchial biopsies and 8 (6 M, 2 F) tumors resected at surgery were analyzed for Cpn by immunohistochemistry (IHC). Males had significantly more often squamous‐cell carcinoma (SCC) than females. Other types of LC were more equally distributed between males and females. The difference between males and females regarding smoking history was significant, and male LC patients had significantly higher levels of IgG and/or IgA antibodies than female LC patients. Male and female LC patients had significantly higher prevalences of high antibody titers than controls. A high prevalence of unusually high titers of specific Cpn antibodies was found in male LC patients. This could indicate that LC may be induced by chronic Cpn infection, since stable high titers of Cpn antibodies, especially IgA, are a hallmark of chronic infections.
Early dissemination of Borrelia burgdorferi without generalized symptoms in patients with erythema migransNote
Authors: OKSI JARMO , MARTTILA HARRI , SOINI HANNA , AHO HEIKKI , UKSILA JAAKKO , VILJANEN MATTI K.
Published: October 2003

The diagnosis of erythema migrans (EM) is not always easy, and reports of culture‐ or PCR‐confirmed diagnosis as well as reports of EM with simultaneous disseminated disease are few. Characteristics and incidence of EM in addition to frequency of early dissemination of B. burgdorferi were studied in the archipelago of South‐Western Finland prospectively using questionnaires, skin biopsies and blood samples. Clinical EM was recognized in 82 patients (incidence 148/100000 inhabitants/year). Of skin biopsy samples, 35.5% were positive by PCR (the majority B. garinii), and 21.5% by cultivation (all B. garinii). Of blood samples, 3.8% were positive by PCR, and 7.7% by cultivation. Of the patients, 30.9% were seropositive at the first visit, and 52.9% 3 weeks later. Of the patients with laboratory confirmed diagnosis, the EM lesion was ring‐like in 31.8% and homogenous in 65.9%. Dissemination of B. burgdorferi, based on culture or PCR positivity of blood samples, was detected in 11.0% of the patients. The frequency of generalized symptoms was nearly the same in patients with as in those without dissemination (22.2% vs 27.4%). Only 21.4% of the patients with culture‐positive EM recalled a previous tick bite at the site of the EM lesion. We conclude that EM lesions are more often homogenous than ring‐like. B. burgdorferi may disseminate early without generalized symptoms.
Urethritis associated with Chlamydia trachomatis: Comparison of leukocyte esterase dipstick test of first‐voided urine and methylene blue‐stained urethral smear as predictors of chlamydial infectionNote
Authors: HEDIN GÖRAN , ABRAHAMSSON GUDRUN , DAHLBERG ERIK
Published: October 2003

The use of nucleic acid amplification tests for the diagnosis of C. trachomatis has made it possible to send urine samples instead of urethral swab specimens to the laboratory. The sensitivity is very high, but not 100%, and we continue to perform a test for urethritis at our STD clinic. The aim of this study was to compare the performance of two alternative tests in the diagnosis of urethritis as predictors of C. trachomatis infection: the leukocyte esterase (LE) dipstick test of first‐voided urine and polymorphonuclear leukocyte counts in a methylene blue‐stained (MBS) urethral smear. Urine samples from 480 male patients attending an STD clinic were analysed using the LE test and LCR assay for C. trachomatis; urethral samples were analysed with MBS urethral smear and LCR. The majority (75.8%) of the 480 patients examined were asymptomatic. Chlamydial infection was detected in 50 patients. The sensitivity, specificity and positive predictive value of the LE test for predicting C. trachomatis infection were 46.0, 91.6 and 39.0%, respectively, among all patients examined and 25.9, 95.8 and 33.3%, respectively, among the asymptomatic patients. The corresponding values for the MBS urethral smear were 76.0, 82.1 and 33.0% among all patients and 63.0, 89.6 and 32.7% among the asymptomatic patients. At our STD clinic we chose to perform the examination of MBS urethral smears in the diagnosis of urethritis because of its higher sensitivity relative to the LE test for predicting C. trachomatis.
Telomerase activity, MIB‐1, PCNA, HPV 16 and p53 as diagnostic markers for cervical intraepithelial neoplasiaNote
Authors: HERBSLEB MALENE , KNUDSEN ULLA B. , ØRNTOFT TORBEN F. , BICHEL PETER , NORRILD BODIL , KNUDSEN AAGE , MOGENSEN OLE
Published: October 2003

The present investigation evaluated the relationship between dysplasia of the uterine cervix and telomerase activity, expression of p53, MIB‐1 and PCNA. Telomerase activity was measured on cervical cytobrush material from 126 women suspected of having dysplasia and 61 controls using the telomeric repeat amplification protocol. Immunohistochemistry was used to detect the tumor suppressor protein p53 and cell proliferation, the latter by MIB‐1 and PCNA expression. Infection with human papillomavirus 16 was detected by PCR amplification and Southern blot hybridization of DNA extracted from the same brush material. Positive telomerase activity was found in 5 of 43 (11.6%) normal samples, 12 of 57 (21.1%) samples with inflammation or koilocytosis, 7 of 17 (41.2%) CIN 1 (cervical intraepithelial neoplasia, grade 1), 8 of 20 (40.0%) CIN 2, and 25 of 42 (59.5%) CIN 3/CIS. Telomerase activity was significantly related to the level of dysplasia (p<0.001) and proliferation measured by MIB‐1 (p=0.019), but not to the level of PCNA (p=0.445), HPV 16 status (p=0.098) or staining for p53 (p=0.271). Dysplasia was also related to PCNA, MIB1, p53, and presence of HPV 16. A sequential increase in the examined parameters, paralleling the progression of abnormality, was observed. PCNA and telomerase showed an increase in CIN 1, MIB‐1 and HPV16 in CIN 2, and finally p53 in CIN 3/CIS.
Abnormal venous structures in salivary gland tumors: vasculature characteristics of pleomorphic adenomaNote
Authors: HIROKAWA MITSUYOSHI , TAMURA MEGUMI , HORIGUCHI HIDEHISA , WAKATSUKI SHINGO , SANO TOSHIAKI
Published: October 2003

To clarify the diagnostic significance of abnormal venous structures present in salivary gland tumors, we examined 21 pleomorphic adenomas, 14 Warthin tumors, 1 oncocytic adenoma, 3 myoepitheliomas, 7 basal cell adenomas, 5 mucoepidermoid carcinomas, and 6 adenoid cystic carcinomas. Verhoeff‐van Gieson staining was carried out and the morphology of the veins within the tumors was observed microscopically. Branching veins, thickened intima of the veins, discontinuous elastic membrane and multilayered elastic membrane were seen in 71.4%, 76.2%, 47.6% and 85.7% of pleomorphic adenomas, respectively, and were abundant and easily found in most cases. The abnormal venous structures were also found in other salivary gland tumors examined, but they were few in number and lacked variety. Elastic fibers extending radially into the surrounding stroma were seen in 66.7% of pleomorphic adenomas, and were not seen in other salivary gland tumors. Our results showed that a variety of abnormal venous structures are more abundant and more easily found in pleomorphic adenoma compared with other salivary gland tumors, and, in particular, that perivascular radiating elastic fibers are characteristic of pleomorphic adenoma. We emphasize that the presence of perivascular radiating elastic fibers may be helpful in diagnosing pleomorphic adenoma in small biopsy specimens.
Stone, paste, shell and metal beads from Sharm
Authors: BARKER DIANE
Published: October 2003

Eighty‐seven beads of various materials are discussed according to the twelve typological groups into which they are divided. This typology is largely based on bead shapes, although manufacturing techniques and materials are also discussed. The beads are also compared to other assemblages in southeastern Arabia.
Polymorphic cytokine genotypes as markers of disease severity in adult periodontitis
Authors: Hendley Thomas M. , Sanders John J. , Palesch Yuko , Pandey Janardan P.
Published: October 2003

Abstract. The distributions of the bi‐allelic interleukin‐1β+3953 and tumor necrosis factor‐σ−308 genotypes were determined in 20 patients with advanced adult periodontitis, 20 patients with plaque associated gingivitis, and 45 referent population subjects. A significant increase in IL‐1β+3953 allele 2 frequency was found in patients with advanced periodontitis compared to referent subjects (the Fisher exact test; p=0.013). Furthermore, the frequency of TNF−σ+308 allele 1 was significantly greater in patients with advanced disease compared to those with plaque associated gingivitis (the Fisher exact test; p=0.014). No significant correlation was observed between genotype and cytokine production in these patient populations.