Apmis

HAHN‐STRÖMBERG, VICTORIA; EDVARDSSON, HENRIK; BODIN, LENNART; FRANZÉN, LENNART

doi: 10.1111/j.1600-0463.2008.00894.xpmid: 18397460

Adhesion proteins are responsible for the structural integrity of epithelial tissue and in tumors this integrity is often lost, resulting in a disorganization of the tissue. In the present study the complexity of the invasive front of colon carcinomas was correlated with cell adhesion protein expression and with polymorphisms in their genes. A complexity index was constructed from 32 colon carcinomas using computer‐assisted morphometry estimating fractal dimension and tumor cell clusters followed by tree analysis. Immunohistochemical staining of beta‐catenin, E‐cadherin, occludin and claudin 2 was used for assessment of protein expression. Genetic screening of tissue from the tumor invasion front with laser microdissection was performed using SSCP and DNA sequencing. Adhesion protein distribution was significantly disturbed in most carcinomas. A single mutation in the gene of beta‐catenin was found but there was no correlation between protein expression and genetic polymorphism. Nor was there any correlation between the complexity of the invasive border and protein distribution or genetic alterations. The results indicate that the complexity of colon carcinoma invasion is not dependent on genetic derangements in the genes of adhesion proteins or the protein distribution. Rather, aberrations in the function of other proteins related to the adhesive proteins could be responsible.

NIKOLAITCHOUK, NATALIA; ANDERSCH, BJÖRN; FALSEN, ENEVOLD; STRÖMBECK, LOUISE; MATTSBY‐BALTZER, INGER

doi: 10.1111/j.1600-0463.2008.00808.xpmid: 18397461

In the present study the lower genital tract microbiota in asymptomatic fertile women (n=34) was identified and quantified by culturing vaginal secretions. Also, vaginal and cervical samples were analyzed by a semiquantitative checkerboard DNA‐DNA hybridization technique (CDH) based on genomic probes prepared from 13 bacterial species (Bacteroides ureolyticus, Escherichia coli, Fusobacterium nucleatum, Gardnerella vaginalis, Mobiluncus curtisii ss curtisii, Prevotella bivia, Prevotella disiens, Prevotella melaninogenica, Atopobium vaginae, Lactobacillus iners, Staphylococcus aureus ss aureus, Streptococcus anginosus, and Streptococcus agalactiae). The bacterial species found by either culture or CDH were correlated with proinflammatory cytokines (IL‐1α, IL‐1β, IL‐6, IL‐8), secretory leukocyte protease inhibitor (SLPI), and endotoxin in the cervicovaginal samples. Grading the women into healthy, intermediate, or bacterial vaginosis (BV) as based on Gram staining of vaginal smears, the viable counts of lactobacilli (L. gasseri) and of streptococci‐staphylococci combined were highest in the intermediate group. In BV, particularly the high concentrations of Actinomyces urogenitalis, Atopobium vaginae, and Peptoniphilus harei were noted (≥1011 per ml). The total viable counts correlated with both cervical IL‐1α and IL‐1β. A strong negative correlation was observed between L. iners and total viable counts, G. vaginalis, or cervical IL‐1α, while it correlated positively with SLPI. Analysis of vaginal and cervical samples from 26 out of the 34 women by CDH showed that anaerobic bacteria were more frequently detected by CDH compared to culture. By this method, A. vaginae correlated with G. vaginalis, and L. iners with S. aureus. With regard to cytokines, B. ureolyticus correlated with both cervical and vaginal IL‐1α as well as with cervical IL‐8, while F. nucleatum, S. agalactiae, S. anginosus, or S. aureus correlated with vaginal IL‐1α. Furthermore, all Gram‐negative bacteria taken together, as measured by CDH, correlated with vaginal endotoxin and inversely with vaginal SLPI. The significance of the results is discussed. In summary, mapping of the identity and quantity of vaginal bacterial species and their association with locally produced host innate immune factors will help in defining various types of abnormal vaginal microbiota, developing new ways of assessing the risk of ascending subclinical infections, and in treating them. CDH appears to be a suitable tool for future analyses of large numbers of clinical samples with an extended number of bacterial probes.

NØRGAARD, METTE; SKRIVER, METTE VINTHER; SØRENSEN, HENRIK TOFT; SCHØNHEYDER, HENRIK CARL; PEDERSEN, LARS

doi: 10.1034/j.1600-0463.2003.11101031.x-i1pmid: 18397462

Few data exist on the risk of miscarriage after exposure to pivmecillinam. We therefore conducted a population‐based case‐control study in a Danish county with 0.5 million inhabitants during the period 1997–2002. We included 1,599 women with a miscarriage recorded in the Hospital Discharge Registry and selected 10 controls per case among primiparous women who had a live birth during the study period. Controls were selected from the Danish Medical Birth Registry. We obtained data on use of pivmecillinam and sulfamethizole from a prescription database. Five cases (0.30%) and 24 controls (0.15%) were exposed to pivmecillinam in the last week before the miscarriage/index date. After adjustment for maternal age, use of antidiabetics or antiepileptics, the odds ratio for miscarriages among users of pivmecillinam compared with non‐users was 2.03 (95% confidence interval: 0.77–5.33) and the corresponding odds ratio for use of sulfamethizole was 1.53 (95% confidence interval: 0.76–3.09). Exposure within 2 to 12 weeks before the miscarriage was not associated with an increased risk. We concluded that use of pivmecillinam was associated with an increased risk of miscarriage, but the risk was not significantly (p=0.64) different from the risk associated with use of sulfamethizole.

ZENG, XIN; XIONG, CHUNRONG; WANG, ZHI; JIANG, LU; HOU, XIAOHUI; SHEN, JUN; ZHOU, MIN; CHEN, QIANMING

doi: 10.1111/j.1600-0463.2008.00741.xpmid: 18397463

Candida albicans carriage has been found to be increased in patients with oral lichen planus. In the present work we have investigated the genotypic profiles of 112 C. albicans strains isolated from patients with erosive or nonerosive OLP, and from healthy controls. The virulence attributes of the isolated strains were compared to elucidate the pathogenetic mechanisms through which C. albicans may cause OLP. We have characterized the genotypic profiles of these isolated strains using a randomly amplified polymorphic DNA assay. In addition, we used assays to measure adhesion to buccal epithelial cells and phospholipase activity to evaluate the virulence attributes of these isolates. Our RAPD analyses revealed four different genotypes, named type A to D, among all isolates, and identified statistically significant associations with disease conditions. Specifically, type A (58.1%) and C (29.0%) were primarily found in erosive OLP, while type A (33.3%) and D (58.3%) were identified in nonerosive OLP. However, the healthy group seemed to have type B (38.5%) and D (61.5%). Using adhesion to BEC assay, we demonstrated that the isolates with genotype A had the strongest adherence among the four genotypes (P=0.000<0.05). The phospholipase activity of the isolates with genotype A and C was higher than for those with genotype B and D (P=0.000<0.05). In conclusion, some C. albicans isolates with special genotypic profiles and virulence attributes may contribute to the pathogenesis of OLP.

VOGEL, T.; VADONIS, R.; KÜHN, J.; EING, B. R.; SENNINGER, N.; HAIER, J.

doi: 10.1111/j.1600-0463.2008.00447.xpmid: 18397464

Anastomotic leakage and septic complications are the most important determinants of postoperative outcome after major surgical resections. Malignant diseases and surgical trauma can influence immune responses and the ability to react against infectious factors, such as bacteria and viruses. Comparable immune suppression can cause viral reactivation in transplantation and trauma patients. In this prospective study, patients who underwent major surgical resections for oesophageal or pancreatic cancer were investigated for the potential involvement of viral reactivation in the development of septic complications. 86 patients (40 oesophageal resections, 27 pancreatic resections, 19 surgical explorations) were included. Viral antigens, viral DNA, antibodies against viral structures (IgG, IgM, IgA) and, in part, viral cultivation were performed for CMV, EBV, HSV1, HSV2, HZV6 and VZV in serum, urine, sputum and swabs from buccal mucosa preoperatively and at postoperative days 1, 3 and 5. Test results were compared with the postoperative outcome (30‐day morbidity, in‐hospital mortality) and clinical scores (SOFA, TISS). For statistical analyses Student's t‐tests and Chi2‐tests were used. The overall complication rate was 19.8% (30‐day morbidity) with an in‐hospital mortality of 1.2% (1/86 patients). Postoperatively, anti‐CMV‐IgG titres were significantly reduced (p<0.05) and remained suppressed in patients with septic complications. Anti‐CMV‐gB‐IgG were also reduced, but showed considerable interindividual differences. Anti‐CMV‐IgA and ‐IgM did not show significant alterations in the postoperative course. In addition, direct viral detection methods did not support viral reactivation in patients in any of the investigated groups. The reduction of anti‐CMV antibodies is likely caused by an immune suppression, specifically by reduced B‐cell counts after major surgical interventions. Viral reactivation, however, did not occur in the early postoperative period as a specific risk for septic complications.

LYTSY, BIRGITTA; SANDEGREN, LINUS; TANO, EVA; TORELL, ERIK; ANDERSSON, DAN I.; MELHUS, ÅSA

doi: 10.1111/j.1600-0463.2008.00922.xpmid: 18397465

Between May and December 2005, 64 multidrug‐resistant isolates of Klebsiella pneumoniae were detected from patients admitted to Uppsala University Hospital. This represented a dramatic increase in ESBL‐producing K. pneumoniae compared to previous years. To investigate the epidemiology and to characterize the resistance mechanisms of the isolates, a study was initiated. Antibiotic susceptibility was determined by means of the Etest and the disc diffusion method. Extended‐spectrum beta‐lactamase (ESBL) production was identified by clavulanic acid synergy test and confirmed with PCR amplification followed by DNA sequencing. DNA profiles of the isolates were examined with pulsed‐field gel electrophoresis (PFGE). All isolates were resistant or exhibited reduced susceptibility to cefadroxil, cefuroxime, cefotaxime, ceftazidime, aztreonam, piperacillin/tazobactam, ciprofloxacin, tobramycin, and trimethoprim‐sulfamethoxazole. They produced ESBL of the CTX‐M‐15 type, and the involvement of a single K. pneumoniae clone was shown. This is the first major clonal outbreak of multiresistant ESBL‐producing K. pneumoniae in Scandinavia. The outbreak demonstrates the epidemic potential of enterobacteria containing ESBLs of the CTX‐M type, even in a country with a relatively low selective pressure and a low prevalence of multiresistant bacteria.

YANG, CHING‐HSIU; HUNG, WEN‐CHUN; WANG, SHENG‐LAN; KANG, WAN‐YI; CHEN, WAN‐TZU; HUANG, YA‐CHUN; SU, YUE‐CHIU; CHAI, CHEE‐YIN

doi: 10.1111/j.1600-0463.2008.00905.xpmid: 18397466

The aim was to investigate the expression of human telomerase reverse transcriptase (hTERT) and cyclin D1 in correlation with clinicopathologic features of urothelial carcinoma (UC). Tissue microarrays (TMA) were constructed from paraffin‐embedded specimens of 94 UC patients and immunohistochemical staining was used. High hTERT expression was found in 50 (53%) of the 94 tumors and was significantly associated with tumor invasiveness and tumor grade (P=0.008 and 0.0190, respectively). High cyclin D1 expression was found in 69 (73%) of the 94 tumors and was significantly associated with non‐invasiveness and smaller tumor size, but there was no correlation with tumor grade. Kaplan–Meier analysis indicated that patients with low hTERT and high cyclin D1 levels had longer local recurrence‐free survival (P=0.0482 and 0.0123, respectively). In addition, patients with high cyclin D1 levels had longer disease‐free survival (P=0.0195). In conclusion, this study demonstrated that hTERT and cyclin D1 may be of recurrence predictive value for UC, thus providing clinicians with ancillary information when deciding on suitable therapeutic strategies in UC.

RAFIEFARD, FARIDEH; ÖRVELL, CLAES; BONDESON, KÅRE

doi: 10.1111/j.1600-0463.2008.00758.xpmid: 18397467

Genotyping of respiratory syncytial (RS) virus group A, by means of a novel method based on PCR, FRET (fluorescence resonance energy transmission) detection and two‐dimensional melting curve analysis, was carried out on 80 RS virus samples of group A collected in Stockholm from 1976 to 2005. The Tm values were assessed for three different genotypes (GA2, GA5 and GA7) circulating in Sweden. Two pairs of probes were used and results of subsequent data analysis were plotted in a two‐dimensional system. The results obtained were compared to genotyping using conventional nucleotide sequencing and phylogenetic tree analysis. It was found that the new assay was able to correctly identify genotype in about 89% of the isolates; it identified the remaining 11% as untypeable and as candidates for conventional nucleotide sequencing. The new method constitutes a complement to nucleotide sequencing and could be useful in studies of large numbers of samples in epidemiological studies.

Karimi, Shirin; Mohammadi, Forouzan; Pejhan, Saviz; Zahirifard, Soheila; Seyfollahi, Leila; Bahadori, Moslem

doi: 10.1111/j.1600-0463.2008.00848.xpmid: 18397468

MORTENSEN, ANOUCK LEUBA; HEEGAARD, STEFFEN; CLEMMENSEN, OLE; PRAUSE, JAN ULRIK

doi: 10.1111/j.1600-0463.2008.00880.xpmid: 18397469

We report the clinical and histopathological characteristics of two cases of signet ring cell carcinoma of the eye lids, and discuss the histogenesis of this neoplasm. Two 72‐year‐old Caucasian males both presented with slowly growing tumours of the eyelids. The tumours were excised and specimens were examined using light‐ and transmission electron microscopic techniques. Clinically, the tumours infiltrated both eyelids on one side of the face with swelling and periocular inflammation, creating a monocle‐like appearance. Extensive clinical work‐up excluded periocular metastases. Histopathologically, the tumours were composed of rather bland cells with mainly histiocytoid morphology. A minor proportion had a signet ring cell appearance. The cytoplasmic inclusions giving the signet ring morphology were PAS‐ and colloidal iron positive. The tumour cells reacted with antibodies against cytokeratins, carcinoembryonic antigen, epithelial membrane antigen, gross cystic disease fluid protein‐15 and lysozyme. Transmission electron microscopy demonstrated tumour cells containing intracytoplasmic vacuoles lined by microvilli. The tumour cells aggregated in duct‐like clusters. A diagnosis of primary signet ring cell carcinoma was made in both cases. Histopathological, immunohistological and ultrastructural findings indicated that the tumours were of sweat gland origin.