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.

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.

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.

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.

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.

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.

The dephosphorylation characteristics of the receptors for epidermal growth factor and platelet-derived growth factor in Swiss 3T3 cell membranes suggest differential regulation of receptor signalling by endogenous protein-tyrosine phosphatases.

Comparison of the phosphotyrosine-specific dephosphorylation of the autophosphorylated receptors for epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) in Swiss 3T3 cell membranes by the endogenous phosphatases revealed striking differences. EGF receptor dephosphorylation was clearly faster than PDGF receptor dephosphorylation and strongly inhibited by Triton X-100 and octylglucoside, whereas PDGF receptor dephosphorylation was to a lesser extent detergent-susceptible. PDGF receptor dephosphorylation was effectively inhibited by phenylarsineoxide, protamine and poly-lysine and partially by N-ethylmaleinimide, whereas EGF receptor dephosphorylation was not affected by these agents. We suggest that these differences in dephosphorylation of EGF and PDGF receptors are due to their differential interaction with membrane-associated protein-tyrosine phosphatases and important for differential regulation of receptor signalling.

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.

Plasma oxytocin but not prostaglandin F2 alpha metabolite levels at cerclage may predict preterm delivery.

Plasma oxytocin and prostaglandin F2 alpha metabolite (PGFM) concentrations were measured in 45 patients admitted for cerclage during the second trimester. Samples were collected before, 3 hours after, and 3 days after the Shirodkar procedure. Uterine activity was recorded by external tocography twice daily for 30 minutes. Twenty-eight women with uncomplicated pregnancy and commensurate gestational age served as controls. Cervical length, measured by ultrasonography, was significantly shorter before cerclage (36 +/- 2 mm) than after cerclage (43 +/- 2 mm) or compared with controls (48 +/- 1 mm). Bishop scores ranged from 3-6 (median 4) in the cerclage group and 0-1 (median 0) in controls. Fifteen cerclage patients and one control delivered preterm 5-22 weeks after the procedure. Initial plasma PGFM levels were significantly higher in cerclage patients than in controls. The cerclage procedure caused an immediate rise in plasma PGFM and a subsequent fall below initial levels to control values. Neither the initial levels of PGFM nor the increments 3 hours after cerclage correlated with the outcome of pregnancy. By contrast, plasma oxytocin levels before cerclage were significantly higher in patients who subsequently delivered preterm than in those who delivered at term. Cerclage resulted in a significant fall in plasma oxytocin at 3 hours in patients with preterm delivery, but after 3 days the oxytocin levels had returned to the precerclage values. Patients who had increased uterine contractions had significantly higher plasma oxytocin levels but lower PGFM levels than those without contractions.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

[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.

[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.

[Vaginal ultrasound follow-up of suspicious smears of the cervix uteri]. / Vaginosonographische Kontrolle bei suspektem Abstrichbefund der Portio uteri.

Using transvaginal sonography (TVS) it is possible to place the scanner probe very close to the organs of the female small pelvis. The near position allows the employment of a higher ultrasound frequency with a better resolving power and thus with more information about the examined structures. A clinical study was performed to investigate whether it is possible to gain some more information about alterations of the uterine cervix surface, after a suspicious cytology result, by applying TVS. A total 59 women with a pathological swab were examined, followed by conisation (using an operating knife) and a D & C. Histological examination showed that in 21 cases with sonographically not suspicious results, 10 cancers in situ and 1 early invasive squamous cell cancer could not be detected. The determined sensitivity of the method was 30.8%; the specificity was 59%. Concluding from the result TVS can be applied only with restriction concerning detection of beginning cancer of the uterine cervix. A statistical significance of vaginosonographical results compared with the suspicious smear results could not be established.

[Vaginal sonography in follow-up of early pregnancies--applications in general practice of established gynecologists]. / Vaginosonographie zur Kontrolle von Frühestschwangerschaften--Einsatz in der Praxis des niedergelassenen Gynäkologen.

In 541 early pregnant women transvaginal sonography (TVS) was performed between 5th and 9th week of gestation. The correct gestational age was verified and information on normal development of the embryo was collected with this routine sonographic control. In one-third of 541 patients disturbances of the pregnancy were found directly or a TVS control was recommended. In 116 cases the gestational age, determined by calculating the period since the last menstruation, had to be corrected by sonographic results. In 12 women ectopic pregnancies without any symptoms were found. Only 47 women of 251 evaluated patients had no risk in pregnancy.

[Vaginal sonographic assessment of a cross section of the amniotic sac in early pregnancy]. / Vaginalsonographische Ermittlung des Fruchtsackquerschnittes in der Frühschwangerschaft.

The present study describes the results of gestation sac measurement in 237 cases in early pregnant women by the technique of transvaginal sonography. Mean distance between uterine cavity and vaginal scanner probe is the reason for early detection of the gestation ring. Comparing with the transabdominal method the gestation sac can be ascertained one week earlier when performing transvaginal sonography, so that a detection of an intrauterine ring-shaped structure is possible to a time when in case of an unknown pregnancy regular menstrual bleeding is expected. Up the 10th week of gestation the amnion sac diameters are constantly smaller than the diameter obtained by transabdominal sonography, from the 11th week on results in both techniques are approximately equal. The early measurement of gestation sac diameter is usefull to determine correct gestation age in case of unknown conception date. From our point of view the gestation ring diameters, obtained by transabdominal technique should not be used father more for pregnancy determination when choosing the transvaginal method.

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.

[Cholesterol granulomatosis of the lungs]. / Cholesteringranulomatose der Lungen.

The cholesterol storage disease of the lung is seldom and therefore we hardly think of it. An evident diagnosis can be given only by histological examinations. 2 cases are described where during life was diagnosed a silicosis and declared as an occupational disease. Autopsy revealed cholesterol storage disease without any signs of silicosis. These findings suggest, that in cases with low occupational quartz-dust-exposure it is advisable to extend the field of diagnostic procedures.

[Differences in proline metabolism of a peptide alkaloid producing and a nonproducing strain of Claviceps purpurea]. / Unterschiede im Prolinstoffwechsel eines Peptidalkaloide-produzierenden und eines nichtproduzierenden Stammes von Claviceps purpurea.

Two strains (pepty 695 and pur 221) of Claviceps purpurea have been used to study the dependence of proline turnover on incubation time (1, 3, 6, and 12 hours) under different culture conditions (synthetic saccharose-citrate medium NL 720 and complex wort medium M 107) by means of tracer technique. In the saprophytically ergotoxine producing strain pepty 695 the proline is utilized in the protein and alkaloid biosyntheses. On the other hand, under non-producing conditions (M 10) the radioactivity of proline-14C is incorporated into many fractions and finally into CO2. In submerged culture (NL 720) the Claviceps strain pur 221 which does not produce ergolines uses proline only to a small extent in the protein biosynthesis, while most of this amino acid is not metabolized. The extent of uptake of proline is not correlated with the alkaloid synthesis. For comparison the turnover of L-glutamate-14C and D,L-ornithine-1-14C by strain pepty 695 cultivated in the NL 720 medium has been studied.