NOX4-driven ROS formation mediates PTP inactivation and cell transformation in FLT3ITD-positive AML cells.

Activating mutations of FMS-like tyrosine kinase 3 (FLT3), notably internal tandem duplications (ITDs), are associated with a grave prognosis in acute myeloid leukemia (AML). Transforming FLT3ITD signal transduction causes formation of reactive oxygen species (ROS) and inactivation of the protein-tyrosine phosphatase (PTP) DEP-1/PTPRJ, a negative regulator of FLT3 signaling. Here we addressed the underlying mechanisms and biological consequences. NADPH oxidase 4 (NOX4) messenger RNA and protein expression was found to be elevated in FLT3ITD-positive cells and to depend on FLT3ITD signaling and STAT5-mediated activation of the NOX4 promoter. NOX4 knockdown reduced ROS levels, restored DEP-1 PTP activity and attenuated FLT3ITD-driven transformation. Moreover, Nox4 knockout (Nox4(-/-)) murine hematopoietic progenitor cells were refractory to FLT3ITD-mediated transformation in vitro. Development of a myeloproliferative-like disease (MPD) caused by FLT3ITD-transformed 32D cells in C3H/HeJ mice, and of a leukemia-like disease in mice transplanted with MLL-AF9/ FLT3ITD-transformed murine hematopoietic stem cells were strongly attenuated by NOX4 downregulation. NOX4-targeting compounds were found to counteract proliferation of FLT3ITD-positive AML blasts and MPD development in mice. These findings reveal a previously unrecognized mechanism of oncoprotein-driven PTP oxidation, and suggest that interference with FLT3ITD-STAT5-NOX4-mediated overproduction of ROS and PTP inactivation may have therapeutic potential in a subset of AML.

Final analysis of the prospective WSG-AGO EC-Doc versus FEC phase III trial in intermediate-risk (pN1) early breast cancer: efficacy and predictive value of Ki67 expression.

BACKGROUND: Taxane-based adjuvant chemotherapy is standard in node-positive (N+) early breast cancer (BC). The magnitude of benefit in intermediate-risk N+ early BC is still unclear. WSG-AGO epiribicine and cyclophosphamide (EC)-Doc is a large trial evaluating modern taxane-based chemotherapy in patients with 1-3 positive lymph nodes (LNs) only. PATIENTS AND METHODS: A total of 2011 BC patients (18-65 years, pN1) were entered into a randomized phase III trial comparing 4 × E90C600 q3w followed by 4 × docetaxel 100 q3w (n = 1008) with the current standard: 6 × F500E100C500 q3w (n = 828) or C600M40F600 d1, 8× q4w (n = 175). Primary end point was event-free survival (EFS); secondary end points were overall survival (OS), toxicity, translational research, and quality of life. Central tumor bank samples were evaluable in a representative collective (n = 772; 40%). Ki-67 was assessed centrally in hormone receptor-positive disease as a surrogate marker for the distinction of luminal A/B-like tumors. RESULTS: Baseline characteristics were well balanced between study arms in both main study and central tumor bank subset. At 59-month median follow-up, superior efficacy of EC-Doc [versus FEC (a combination of 5-fluorouracil, epirubicin, and cyclophosphamide)] was seen in EFS and OS: 5-year EFS: 89.8% versus 87.3% (P = 0.038); 5-year OS: 94.5% versus 92.8% (P = 0.034); both tests one-tailed. EC-Doc caused more toxicity. In hormone receptor-positive (HR)+ disease, only high-Ki-67 tumors (≥ 20%) derived significant benefit from taxane-based therapy: hazard ratio = 0.39 (95% CI 0.18-0.82) for EC-Doc versus FEC (test for interaction; P = 0.01). CONCLUSION: EC-Doc significantly improved EFS and OS versus FEC in intermediate-risk BC (1-3 LNs) within all subgroups as defined by local pathology. In HR+ disease, patients with luminal A-like tumors may be potentially over-treated by taxane-based chemotherapy. CLINICAL TRIAL NUMBER: ClinicalTrials.gov, NCT02115204.

Role of endorectal prostate MRI in patients with initial suspicion of prostate cancer.

PURPOSE: To evaluate the role of conventional endorectal prostate MRI in patients with initial suspicion of prostate cancer. MATERIALS AND METHODS: Ethics board approval was received for this retrospective study of 87 men who underwent 1.5-Tesla conventional prostate MRI with a combination of endorectal and body phased-array coils for suspected prostate cancer before their first systematic 12-core TRUS-guided biopsy. Three radiologists independently analyzed the images, dividing the prostate into 12 regions corresponding to the biopsy scheme and scoring each region for the presence of prostate cancer on a 5-point scale. Results were analyzed by prostate region. ROC analysis was done and descriptive statistics were calculated. The negative predictive value, specificity, sensitivity and positive predictive value were calculated using dichotomized scores (benign tissue = scores of 1 and 2; malignant tissues = scores of 3, 4, and 5). RESULTS: Biopsy revealed cancer in 47/87 patients (26 low-grade [Gleason score 6]; 21 high-grade [Gleason score ≥ 3 + 4]), and 184/1044 cores (77 low-grade and 107 high-grade) with a median of 3 positive cores per cancer patient (range 1 - 12). The areas under ROC curves were 0.65 - 0.67 for cancer detection by region overall and 0.75 - 0.76 for the detection of high-grade cancer by region. Statistic figures for the detection of all cancers/high-grade cancers by region were as follows: negative predictive value, 87.4 - 88.2 %/92.6 - 93.1 %; specificity, 72.3 - 79.4 %/71.5 - 79.8 %; sensitivity, 49.5 - 54.8 %/62.6 - 69.2 %; and positive predictive value, 29.3 - 34.0 %/29.4 - 34.7 %. CONCLUSION: In patients with suspected prostate cancer, negative MRI findings indicate the absence of high-grade prostate cancer on subsequent TRUS-guided 12-core biopsy with high probability. However, agreement between conventional 1.5-T endorectal prostate MRI and systematic 12-core TRUS-guided biopsy for the detection of prostate cancer appears to be moderate.

Role of endorectal prostate MRI in patients with initial suspicion of prostate cancer.

PURPOSE: To evaluate the role of conventional endorectal prostate MRI in patients with initial suspicion of prostate cancer. MATERIALS AND METHODS: Ethics board approval was received for this retrospective study of 87 men who underwent 1.5-Tesla conventional prostate MRI with a combination of endorectal and body phased-array coils for suspected prostate cancer before their first systematic 12-core TRUS-guided biopsy. Three radiologists independently analyzed the images, dividing the prostate into 12 regions corresponding to the biopsy scheme and scoring each region for the presence of prostate cancer on a 5-point scale. Results were analyzed by prostate region. ROC analysis was done and descriptive statistics were calculated. The negative predictive value, specificity, sensitivity and positive predictive value were calculated using dichotomized scores (benign tissue = scores of 1 and 2; malignant tissues = scores of 3, 4, and 5). RESULTS: Biopsy revealed cancer in 47/87 patients (26 low-grade [Gleason score 6]; 21 high-grade [Gleason score ≥ 3 + 4]), and 184/1044 cores (77 low-grade and 107 high-grade) with a median of 3 positive cores per cancer patient (range 1 ­ 12). The areas under ROC curves were 0.65 ­ 0.67 for cancer detection by region overall and 0.75 ­ 0.76 for the detection of high-grade cancer by region. Statistic figures for the detection of all cancers/high-grade cancers by region were as follows: negative predictive value, 87.4 ­ 88.2 %/92.6 ­ 93.1 %; specificity, 72.3 ­ 79.4 %/71.5 ­ 79.8 %; sensitivity, 49.5 ­ 54.8 %/62.6 ­ 69.2 %; and positive predictive value, 29.3 ­ 34.0 %/29.4 ­ 34.7 %. CONCLUSION: In patients with suspected prostate cancer, negative MRI findings indicate the absence of high-grade prostate cancer on subsequent TRUS-guided 12-core biopsy with high probability. However, agreement between conventional 1.5-T endorectal prostate MRI and systematic 12-core TRUS-guided biopsy for the detection of prostate cancer appears to be moderate.

[Nuchal translucency measurement--an effective method for early recognition of fetal disease?]. / Nackentransparenz-Messung--eine effektive Untersuchung zur Früherkennung fetaler Erkrankungen?

BACKGROUND: In recent years, an increasing amount of experience has been collected in measuring the nuchal translucency (NT) of the fetus in early pregnancy. While all fetuses develop a measurable collection of fluid in the area of the neck between the 11th and 14th weeks of pregnancy, the fact that fetuses with chromosomal disorders, cardiac defects, and syndromal diseases, in particular, reveal an above-average incidence of increased NT has been noticed. OBJECTIVE: By processing our own patient data from the past two years, we intend to elucidate the question of whether NT measurement is a sensible investigation for the early detection of fetal problems. PATIENTS AND METHOD: NT measurements were carried out in 199 fetuses; these measurements were standardized according to the guidelines of the Foetal Medicine Foundation in London, in whose multicentric study we are participating. The patients were under the care of our prenatal diagnosis and therapy department and were referred to us from external sources. RESULTS: NT within the reference range was determined in 152 fetuses; NT exceeded the reference value in 47 fetuses. Of those fetuses with increased NT, 7 fetuses revealed a chromosomal anomaly, 1 foetus was suffering from a cardiac defect, 3 fetuses were suffering from other organ abnormalities, and 3 fetuses were determined to be suffering from syndromal disease. None of the fetuses whose NT measurements were within the reference range was discovered to be suffering from any of the above-mentioned problems. CONCLUSION: Even this relatively small group of patients reveals that NT measurement is a very effective filter for detecting certain fetal diseases during the early fetal period.

[Empirical report on acute twin-twin transfusion syndrome with single intrauterine death]. / Erfahrungsbericht über ein akutes feto-fetales Transfusionssyndrom mit letalem Ausgang.

We report on a case of acute twin-twin transfusion syndrome in a twin pregnancy in the 26 th under primarily unclear conditions. On admission to the hospital, one fetus was not showing signs of life anymore, while the Doppler indices and CTG of the living fetus showed signs of acute distress. On the scan both fetuses showed adequate and symmetric growth as well as symmetric and normal amniotic fluid amounts, indicating a lack of typical signs for chronic twin-twin transfusion syndrome. The emergency cesarean section performed under the assumption of acute twin-twin transfusion syndrome, which unfortunately could not save the second twin, confirmed our suspected diagnosis.

Bis(1H-2-indolyl)-1-methanones as inhibitors of the hematopoietic tyrosine kinase Flt3.

Aberrant expression and activating mutations of the class III receptor tyrosine kinase Flt3 (Flk-2, STK-1) have been linked to poor prognosis in acute myeloid leukemia (AML). Inhibitors of Flt3 tyrosine kinase activity are, therefore, of interest as potential therapeutic compounds. We previously described bis(1H-2-indolyl)-1-methanones as a novel class of selective inhibitors for platelet-derived growth factor receptors (PDGFR). Several bis(1H-2-indolyl)-1-methanone derivatives, represented by the compounds D-64406 and D-65476, are also potent inhibitors of Flt3. They inhibit proliferation of TEL-Flt3-transfected BA/F3 cells with IC(50) values of 0.2-0.3 microM in the absence of IL-3 but >10 microM in the presence of IL-3. Ligand-stimulated autophosphorylation of Flt3 in EOL-1 cells and corresponding downstream activation of Akt/PKB are effectively inhibited by bis(1H-2-indolyl)-1-methanones whereas autophosphorylation of c-Kit/SCF receptor or c-Fms/CSF-1 receptor is less sensitive or insensitive, respectively. Flt3 kinase purified by different methods is potently inhibited in vitro, demonstrating a direct mechanism of inhibition. 32D cells, expressing a constitutively active Flt3 variant with internal tandem duplication are greatly sensitized to radiation-induced apoptosis in the presence of D-64406 or D-65476 in the absence but not in the presence of IL-3. Thus, bis(1H-2-indolyl)-1-methanones are potential candidates for the treatment of Flt3-driven leukemias.

[Typical forms of choriocarcinoma in clinical practice--diagnosis and therapeutic course in four patients]. / Typische Erscheinungsformen des Chorionkarzinoms in der klinischen Praxis--Diagnosestellung und therapeutischer Verlauf bei vier Patientinnen.

The paper reports on four patients with choriocarcinoma. In two of them, the choriocarcinoma was found after abortion, in one of them following termination of pregnancy, and in the last patient a hydatidiform mole was present. In all patients increased beta-HCG was found. One patient had lung metastasis at the time of diagnosis. In another patient, choriocarcinoma was suspected owing to ultrasonographic vaginal examination. According to the Bagshawe Score, 3 patients were low-risk and were subjected to methotrexate. One patient was medium-risk and received PEB chemotherapy. All four patients are regarded as cured.

[Experiences of the elderly during hospital stay]. / Erfahrungen alter Menschen mit einem Krankenhausaufenthalt.

In Germany the current population structure is characterized by a high proportion of elderly people. This proportion will continue to rise in the future. With the aging of the population the morbidity and the need for ambulatory and, in particular, stationary health care will increase. In order to ensure an appropriate supply for the elderly, it is, among other things necessary to consider the older patients' subjective positive and negative experiences with the health care system. In this paper, the experiences of elderly patients with hospitalisation are presented. In four hospitals 123 patients at the age of 59 to 90 years were asked about their experiences with the stay. For this study a new instrument was developed and applied. Practicability of the new instrument, an oral interview, could be demonstrated. Patients expressed predominantly positive experiences, some aspects of care, however, were judged critically. Furthermore, in the experiences of older patients significant sex-specific differences were seen. Age-specific and disease-specific differences were less pronounced.

Patient with trisomy 6 mosaicism.

Trisomy 6 and trisomy 6 mosaicism were found in chorionic villi cell culture and short term incubation in a prenatal diagnosis at 12 weeks of gestation in a pregnancy with a growth retarded fetus showing nuchal translucency. The child was born in the 25th gestational week with a number of malformations including heart defects, deep-set ears, cleft right hand, cutaneous syndactylies, and overlapping toes of irregular shape and length. Trisomy 6 was not found in peripheral blood lymphocytes but was confirmed in umbilical cord fibroblasts. Currently, at the age of 2-3/4 years, the development of the child is relatively normal despite considerable growth delay. At the age of two years, she developed a papular erythema clinically suggestive of epidermal nevi. Cytogenetic analysis of fibroblast cultures derived from skin from a right hand finger and the inguinal area confirmed the presence of a trisomy 6 mosaicism. This is the first observation of a liveborn with trisomy 6 mosaicism.

Negative regulation of Ros receptor tyrosine kinase signaling. An epithelial function of the SH2 domain protein tyrosine phosphatase SHP-1.

Male "viable motheaten" (me(v)) mice, with a naturally occurring mutation in the gene of the SH2 domain protein tyrosine phosphatase SHP-1, are sterile. Known defects in sperm maturation in these mice correlate with an impaired differentiation of the epididymis, which has similarities to the phenotype of mice with a targeted inactivation of the Ros receptor tyrosine kinase. Ros and SHP-1 are coexpressed in epididymal epithelium, and elevated phosphorylation of Ros in the epididymis of me(v) mice suggests that Ros signaling is under control of SHP-1 in vivo. Phosphorylated Ros strongly and directly associates with SHP-1 in yeast two-hybrid, glutathione S-transferase pull-down, and coimmunoprecipitation experiments. Strong binding of SHP-1 to Ros is selective compared to six other receptor tyrosine kinases. The interaction is mediated by the SHP-1 NH(2)-terminal SH2 domain and Ros phosphotyrosine 2267. Overexpression of SHP-1 results in Ros dephosphorylation and effectively downregulates Ros-dependent proliferation and transformation. We propose that SHP-1 is an important downstream regulator of Ros signaling.

[Pronounced bilateral mammary hypertrophy as a rare manifestation of acute lymphatic leukemia]. / Ausgeprägte bilaterale Mammahypertrophie als ungewöhnliche Manifestation einer akuten lymphatischen Leukämie.

A 33-year-old patient, hitherto healthy, has been admitted for clarification of bilateral mammary hypertrophy. In the course of the usual routine examination a lump of the size of a cherry was identified in the right breast. Within 6 weeks both breasts had become tight and grown symmetrically to three times their original size. A provisionary diagnosis of high malignant non-hodgkin lymphoma was made by multiple high speed needle biopsies and was later confirmed by a surgical tissue specimen of the right breast. For further classification a bone-marrow biopsy was taken from the pelvic bone and an immature acute lymphoblastic leukemia (ALL), known as preB1-ALL, diagnosed. Sonographic examination, computer tomography of the thorax and mammographical findings, as well as symptoms, outcome and differential diagnosis of a proliferative lymphatic disease with first manifestation in both breasts are presented and discussed.

Does texture analysis improve breast ultrasound precision?

OBJECTIVE: To evaluate the possibility of distinguishing between benign and malignant breast tumors using a computer-aided evaluation of echogenicity and echostructure of ultrasound findings at certain focal points. STUDY DESIGN: The ultrasound images from 89 cases of breast tumor were documented under standardized conditions using a linear array machine and 7.5 MHz transducer. In each sonographic image, the maximum area of the 'region of interest' of the tumor was marked and then subjected to consecutive statistical analysis and correlation with the histological findings. For evaluation of tumor status eight parameters of first and second order texture statistics (gray level histogram, Fourier analysis, co-occurrence matrix) were applied. RESULTS: Benign tumors were clearly distinguished from carcinomas in the evaluation of the co-occurrence matrix and the Fourier analysis on the basis of Wilcoxon and Student t-test (P < 0.05) but not in the gray level histogram. Using logistic regression a sensitivity of 73.8% and a specificity of 54.2% were obtained. A statistically significant difference between benign tumors and moderately differentiated together with poorly differentiated carcinomas could be demonstrated. CONCLUSION: This study concludes that texture analysis appears to distinguish between benign and most malignant tumors. A computer texture analyzing system is able to improve the subjective assessment of ultrasound images of the breast but can not replace it. Where the limits of subjective assessment of a given tumor are reached, computerized texture analysis will provide additional information in the differentiation of benign from malignant findings.

Perinuclear localization of the protein-tyrosine phosphatase SHP-1 and inhibition of epidermal growth factor-stimulated STAT1/3 activation in A431 cells.

The SH2 domain protein-tyrosine phosphatase SHP-1 has been shown earlier to bind to the epidermal growth factor receptor and to have the capacity for receptor dephosphorylation. New bi- and tricistronic expression vectors (pNRTIS-21 and pNRTIS-33, respectively) based on the tetracycline system were constructed and employed to generate stable cell lines with inducible expression of SHP-1. Inducible overexpression of SHP-1 in A431 cells led to attenuation of epidermal growth factor (EGF) receptor autophosphorylation and of EGF-induced DNA binding of 'signal transducers and activators of transcription' (STAT) 1 and 3. SHP-1 was localized in the cytoplasm with an enrichment in the perinuclear compartment. Association of SHP-1 with perinuclear structures may form the basis for a partial cofractionation with nuclei observed in different types of transfected cells and also with endogenous SHP-1 in U-937 cells. Treatment of SHP-1-overexpressing A431 cells or of HaCaT human keratinocytes expressing SHP-1 endogenously with the Ca2+-ionophore A23187 resulted in partial nuclear accumulation of SHP-1. Thus, SHP-1 may interact with substrates or regulatory proteins in perinuclear or nuclear structures.

[Terminal CTG as an expression of high degree fetal distress--1 case reports]. / Das terminale CTG als Ausdruck einer hochgradigen Gefährdung des Kindes--zwei Kasuistiken.

The cases of two patients, in whom conspicuous CTGs with restricted oscillation and late decelerations were registered in the final trimester of pregnancy, are presented. Following immediate hospitalisation and the rapid execution of a caesarean section, two depressed, severely acidotic neonates were born. Whilst the course of the pregnancy in the first case had been completely inconspicuous up to that point, and the acute occurrence of placental insufficiency must be assumed, the second patient was subject to pregnancy-induced hypertension with discrete foetal growth retardation. It is shown that two almost identical pathological CTG registrations may have different causes but that one must assume a high degree of sub partu risk to the child on the occurrence of a terminal CTG witch is characterised by line-shaped oscillation, possibly in combination with late decelerations.

[Efficacy of ultrasound screening in pregnancy]. / Qualität des Ultraschall-Screenings in der Schwangerenvorsorge.

OBJECTIVE: The purpose of this study was to examine the effectiveness of ultrasound screening in pregnancy. MATERIAL AND METHODS: Therefore, it was registered whether fetal malformations in a study population of 11,172 deliveries were already diagnosed before birth. RESULTS: 341 defects were found in 297 children from mothers who had had prenatal care. Most anomalies were seen in the urogenital tract (n = 98; 28.7%), the heart (n = 67; 19.6%), the connective tissue (n = 39; 11.4%), the gastrointestinal tract (n = 32; 9.4%), and in the central nervous system (n = 33; 9.7%). Chromosomal anomalies (n = 22; 6.5%) and orofacial defects (n = 21; 6.2%) were more rare. 8.8% of all defects were lethal, 37% severe. 237 (69.5%) were classified as "diagnosable by ultrasound prenatally". 125 of them (53%) were identified prenatally, with high rates of 71% in central nervous system, 65.5% in intestinal and 54% in urogenital tract, while the detection rate was only 13.6% in chromosomal and 3.3% in cardiac defects. Only 14.3% were found before 24 weeks of gestation. CONCLUSIONS: Thus, the effectiveness of ultrasound screening has to be improved by adequate measures.

Oxidation of phenanthrene by a fungal laccase in the presence of 1-hydroxybenzotriazole and unsaturated lipids.

Phenanthrene, a polycyclic aromatic hydrocarbon, was efficiently oxidized by laccase in the presence of both 1-hydroxybenzotriazole and unsaturated lipids. 73% of initially added phenanthrene was degraded within 182 hours to give phenanthrene-9,10-quinone and 2,2'-diphenic acid as the major products. The system was also able to peroxidize linoleic acid to its corresponding hydroperoxides suggesting the involvement of lipid peroxidation in laccase catalyzed phenanthrene oxidation. Lipid peroxidation by laccase required 1-hydroxybenzotriazole and did not depend on Mn2+ and H2O2 suggesting that the chemical reactions involved differ from those previously reported for manganese peroxidase.

Activation of platelet-derived growth factor (PDGF) receptor dephosphorylation in intact Swiss 3T3 cells by elevators of intracellular Ca2+ and cAMP.

We investigated the modulation of platelet-derived growth factor (PDGF) receptor dephosphorylation in Swiss 3T3 cells using a novel assay permitting monitoring of receptor dephosphorylation in intact cells. PDGF treatment of the cells reduced the receptor dephosphorylation rate to 41%, the elevators of intracellular Ca2+, A23187 and thapsigargin increasing it to 227 and 138%, respectively. The cAMP elevators forskolin and isobutylmethylxanthine also accelerated PDGF receptor dephosphorylation. The involvement of Ca(2+)- and cAMP-dependent protein kinases in the regulation of PDGF receptor dephosphorylation is suggested.

[Texture analysis: a new method for evaluating ultrasound imaged lesions of the breast]. / Texturanalyse: Ein neues Verfahren zur Beurteilung sonographisch darstellbarer Herdbefunde der Mamma.

Echogenicity and echostructure in sonomammography are essential criteria to determine the malignancy of a breast tumor. Looking for a correlation between echostructure of an ultrasound image and its histopathology a texture analysis was performed. Ultrasonic examinations were carried out by means of a 7.5-MHz linear scanner under standardized conditions. 71 mammasonographic findings were documented by video tape, each tumor in 2 dimensions. In a second step 'the region of interest', that means the whole tumor area, was marked and statistically analyzed by a dedicated computer system. The values of 23 first- and second-order statistical texture parameters were compared with the histopathology of the tumor. They include grey level histogram, Fourier analysis and cooccurrence matrix. The most important results were obtained in the histogram analysis. An obvious distinction could be worked out concerning carcinomas, adenomas, mastopathic tumors, and fat necrosis. Except the difference between carcinomas and adenomas this distinction was statistically significant. Fourier analysis and coocurrence matrix significantly separate fibrocystic lesions, fat necrosis, scars, and cystic structures from malignant tumors. Furthermore the maximum value method was used to compare the different parameters with regard to their capability of discriminating benign and malignant tumors. The extreme values of 100% were mostly observed in the case of seromas and scars. A significant difference was found by histological grading of carcinomas, too. Highly differenciated carcinomas show capability of results comparable to those of adenomas. By means of a computer texture analysis program it is possible to find a correlation between echostructure and histopathology of breast tumors.(ABSTRACT TRUNCATED AT 250 WORDS)