Expression of MRP1 and related transporters in human lung cells in culture.

Multidrug resistance type 1 P-glycoproteins (P-gp) and multidrug resistance associated proteins (MRP) were studied in differentiated primary human lung cells in culture, in comparison with permanent human lung cell lines and primary alveolar type II cells from rat lung. AII cells exhibited low basal levels of mdr1b mRNA, that increased over time and after oxygen radical production induced by paraquat. mRNAs coding for antioxidative enzymes catalase (CAT), maganese superoxide dismutase (Mn-SOD) and copper/zinc superoxide dismutase (Cu/Zn-SOD) were not changed. H358, A549, H322 cells expressed low levels of MDR1 mRNA, but the mdr1 substrate rhodamine 123 (Rh 123) was transported out of H358 and H322 cells in a non-invasive, single cell fluorescence assay. The dye efflux could be inhibited by the chemosensitizer, verapamil. Normal human bronchial epithelial cells (NHBEC) expressed immuno-reactive MDR1 P-gp and the MPR protein that was active in the fluorescence assay using the MRP substrate carboxy-dichlorofluorescein (CDF) and MK-571 as an inhibitor. We did observe inter-individual variation of MRP in both the mRNA and the immunoreactive protein in NHBEC culture. Over time (12 weeks) the protein was relatively stable in NHBEC and epithelial cells from peripheral lung (PLC), but the mRNA level was drastically increased when explant cultures were continued (18 weeks).

Microscopic (R1) and macroscopic (R2) residual disease in patients with resected non-small cell lung cancer.

OBJECTIVES: This retrospective study evaluates the probability of survival in patients who had undergone resection for non-small cell lung cancer (NSCLC) and in whom residual disease at the resection margins was found. METHODS: During a period of 6 years, 596 patients with NSCLC were operated upon with curative intention. Residual disease at the resection margin was divided into microscopic (R1) and macroscopic (R2). RESULTS: Twenty-six patients (4.4%) showed R1 and 12 (2%) R2 residual disease. An extrabronchial (thoracic wall, vessels) R1 situation was found in five patients and a bronchial R1 infiltration in 21 cases. The bronchial resection margin was subject to peribronchial infiltration in most cases (16/21). A total of 17/21 (65%) patients with bronchial infiltration had N2 disease. Thirty day lethality was 3.8% in the R1 group. Fifteen patients had postoperative irradiation. The 5-year survival rate for patients with R1 resection was 14%. The differences in survival between patients with extrabronchial vs. bronchial infiltration and N0/N1 vs. N2 were significant using univariate analysis. Adjuvant radiation did not result (especially in N2 disease) in a survival benefit. Among 12 patients with macroscopic residual disease (R2), 3/12 (25%) died within the first 30 days after the operation, and none of the R2 patients survived the first year after the operation. CONCLUSIONS: Patients with an R1 situation have a survival rate of 14% comparable to curative resected patients (RO) in stage III. Adjuvant radiation had no clear effect on survival. Patients with macroscopic tumor (R2) should receive palliative treatment after the operation depending on their condition.

Cerebral heterotopia appearing as an extranasal polyp.

Cerebral heterotopia represents a rare tumorlike lesion related in its development to the encephaloceles but differing from these by non-connection to the cranial cavity. We describe an unusual case of cerebral heterotopia appearing as a pedunculated polyp on the upper lip of a newborn. As the distinction between encephaloceles and cerebral heterotopias is based on the correlation of radiologic and clinical findings, the surgical treatment of tumorlike lesions of the nasal and pharyngeal region in children requires a careful preoperative evaluation of their possible connection to the cranial cavity.

[Prolonged course of tick-borne ulceroglandular tularemia in a 20-year-old patient in Germany--case report and review of the literature]. / Prolongierter Verlauf einer durch Zeckenstich übertragenen ulzeroglandulären Tularämie bei einer 20-jährigen Patientin.

HISTORY AND ADMISSION FINDINGS: A 20-year-old female patient presented with painful axillary lymphadenopathy. She reported a tick bite five months ago in her right hand followed by fever, chills and regional lymphadenopathy. Empiric antibiotic treatment with doxycyclin and ciprofloxacin had led to defervescence but no change in painful lymph node swellings. Surgical removal of a cubital lymph node had already been performed three months after the tick bite. INVESTIGATIONS: Laboratory findings were normal except for moderate elevation of C-reactive protein. Serology confirmed the suspected clinical diagnosis of ulceroglandular tularemia. Retrospective real-time PCR (markers fopA and tul4) for Francisella tularensis from the previously removed lymph node (paraffin tissue blocks) was negative. DIAGNOSIS, TREATMENT AND COURSE: Clinical presentation and serological test results were consistent with a prolonged course of tick-borne ulceroglandular tularemia associated with reactive lymph node swelling. The patient requested surgical removal of the painful axillary lymph node. Histology showed reticulocytic, abscess forming lymphadenitis with pseudotuberculosis type of granulomatosis and negative acid-fast staining. Blood culture, capture ELISA and real-time PCR for Francisella tularensis performed on material from the lymph node preparations were negative. A complete recovery was achieved without renewed antibiotic treatment. CONCLUSIONS: According to recent seroprevalence studies, the emergence of tularemia as a rare zoonosis in Germany is clinically underestimated. This case report illustrates possible appearance of the disease in other than known risk groups (e.g. hunters, lumbermen). Ectoparasites like infected ticks have to be considered as vectors, even in non-endemic regions.

[Histopathology after implantation of lid weights]. / Histopathologie nach Implantation von Lidgewichten.

BACKGROUND: Various implants are used to treat lagophthalmos in facial paralysis. The implantation of gold or platinum eyelid weights (lid loading) is being used increasingly. Histological changes have not yet been systematically examined in the implant capsule during explantation of the eyelid weight. MATERIAL AND METHODS: Of 26 explanted implants, 18 implants in 17 patients were followed-up using light microscopy from 1997 to 2004. We semi-quantitatively evaluated the density of the inflammatory infiltrate and recorded the occurrence of foreign bodies in the tissue. The inflammatory infiltrate was also immunohistochemically examined to characterize cellular composition. RESULTS: In the semi-quantitative examination of inflammatory infiltration density, 60% of the gold implants showed a moderately dense or dense inflammatory infiltrate, while there was moderately dense infiltration in only 12.5% of the platinum implants. The immunohistochemical examination revealed a mixed lymphocytic inflammatory infiltrate which tended to a preponderance of B lymphocytes (humoral response). This is typical of unspecific chronic inflammation. CONCLUSIONS: Both gold and platinum are suitable for treating lagophthalmos, however, platinum has a higher density and better biocompatibility than gold.

[Morphological variability of synovial sarcoma in childhood]. / Morphologische Variabilität des synovialen Sarkoms im Kindesalter.

We report on an unusual locally recurrent plantar soft tissue sarcoma in a 7-years-old boy. Due to an unusual morphology a clear diagnosis was initially not possible. The histologically different recurrent tumor was classified as synovial sarcoma by means of immunohistochemistry, ultrastructural and molecular analysis. This case demonstrates the variable morphological appearance of synovial sarcoma and the value of modern diagnostic procedures in such circumstances.

[Malignant epithelioid hemangioendothelioma of the liver - a very rare tumor in children]. / Das maligne epitheloide Hämangioendotheliom der Leber. Ein sehr seltener Tumor im Kindesalter.

We report on a 12-year old boy suffering from malignant epithelioid hemangioendothelioma of the liver, which is a very rare tumor in childhood. The tumor was detected by ultrasound examination at the age of 10 and appeared at that time as a solitary intrahepatic nodular lesion. During the following 2 years multiple nodular lesions developed in both hepatic lobes. There were neither any suspect anamnestic findings nor abnormal clinical or laboratory data. The tumor showed the typical histomorphological, immunohistochemical, and ultrastructural features of this entity, which is usually seen in older patients. We investigated proliferative activity, apoptotic regulation, and expression of VEGF and VEGF-receptor flk-1 by means of immunohistochemical techniques. According to the known slow growth activity of these tumors we found only a few Ki-67 positive tumor cells. We did not detect any apoptotic cells using TUNEL technique. The positive immunoreaction of the tumor cells with antibodies against VEGF and VEGF-receptor flk-1 may indicate the regulation of tumor growth by angiogenetic factors. We present our findings together with a summary of the most important publications of recent years concerning these tumors.

Expression of cytochrome P450 2E1 in normal human bronchial epithelial cells and activation by ethanol in culture.

Serum-free primary cultures of human bronchial epithelial cells and freshly isolated samples of human bronchial epithelium were used to investigate basal expression of the cytochrome P450 enzyme CYP2E1 and its activation or induction by ethanol in bronchial epithelial cells. The cultures consisted of > or =95% cells of epithelial characteristics as determined by transmission electron microscopy and immunohistochemical staining. Monolayers were obtained from explants over a period of several months via transfer of tissue into new dishes ('generations'1-5). Using RT-PCR analysis, basal expression of mRNAs coding for CYP2B7, CYP2F1 and CYP2E1 were detected in cultures from several donors. The basal expression of CYP2E1 protein and mRNA showed differences between the donors. The mRNA was detected even in cultures from higher generations and increased in some cultures over time. The CYP2E1 protein content was low and in most cultures of generations 2-5 could not be detected by immunoblot analysis of native protein extracts. Nevertheless, in some cases immunoreactive CYP2E1 protein was present in monolayers obtained from the fourth and fifth transfer (18-week 'generation'). CYP2E1 activity was measured via 6-hydroxylation of chlorzoxazone either by a destructive assay using cell lysate or by a non-invasive assay using the medium of cell cultures. In short-term cultured isolated bronchial epithelium, ethanol treatment increased CYP2E1 activity by up to 5-fold within 4 days but with inter-individual differences. In cells up to 4 weeks in culture, CYP2E1 activity remained inducible by a single dose of ethanol. Differentiated primary human cells in culture may be useful tools as model systems for the evaluation of CYP2E1-driven processes in man.