Breast conserving surgery in combination with intraoperative radiotherapy after previous external beam therapy: an option to avoid mastectomy.

PURPOSE: Mastectomy is the standard procedure in patients with in-breast tumor recurrence (IBTR) or breast cancer after irradiation of the chest due to Hodgkin's disease. In certain cases a second breast conserving surgery (BCS) in combination with intraoperative radiotherapy (IORT) is possible. To date, data concerning BCS in combination with IORT in pre-irradiated patients are limited. This is the first pooled analysis of this special indication with a mature follow-up of 5 years. METHODS: Patients with IBTR after external beam radiotherapy (EBRT; treated in two centers) for breast cancer were included. Patients with previous EBRT including the breast tissue due to other diseases were also included. IORT was performed with the Intrabeam™-device using low kV X-rays. Clinical data including outcome for all patients and toxicity for a representative cohort (LENT-SOMA scales) were obtained. Statistical analyses were done including Kaplan-Meier estimates for local recurrence, distant metastasis and overall survival. RESULTS: A total of 41 patients were identified (39 patients with IBTR, 2 with Hodgkin`s disease in previous medical history). Median follow-up was 58 months (range 4-170). No grade 3/4 acute toxicity occurred within 9 weeks. Local recurrence-free survival rate was 89.9% and overall survival was 82.7% at 5 years. Seven patients developed metastasis within the whole follow-up. CONCLUSIONS: BCS in combination with IORT in IBTR in pre-irradiated patients is a feasible method to avoid mastectomy with a low risk of side effects and an excellent local control and good overall survival.

Use of the fluorescent dye tetramethylrhodamine methyl ester perchlorate for mitochondrial membrane potential assessment in human spermatozoa.

Mitochondrial membrane potential (ΔΨm) is an indicator of sperm quality and its evaluation complements the standard semen analysis. The fluorescent dye JC-1 has been widely used to assess sperm ΔΨm; however, some problems have been detected under certain experimental conditions. Another fluorescent compound, tetramethylrhodamine methyl ester perchlorate (TMRM), has been used in somatic cells and bovine spermatozoa but not in human spermatozoa. TMRM accumulates in hyperpolarised mitochondria and the fluorescence intensity of this compound correlates with ΔΨm. Thus, the aim of this study was to evaluate and validate the usefulness of the fluorescent dye TMRM for measuring sperm ΔΨm. The results showed that TMRM accurately detects sperm populations displaying either high or low ΔΨm. Moreover, TMRM was able to measure sperm ΔΨm under the experimental conditions in which JC-1 had previously presented difficulties. Differences in ΔΨm according to sperm and semen quality were properly detected and a positive correlation between ΔΨm and conventional semen parameters was observed. Finally, a positive correlation was found between the ΔΨm measurement by TMRM and by the widely used JC-1. In conclusion, TMRM is a simple, time-effective method, easy to set in laboratories equipped with flow cytometry technology, and can accurately detect changes in ΔΨm with efficiency comparable to JC-1 without its limitations.

Cryopreservation of whole ovine ovaries with pedicles as a model for human: parameters of perfusion with simultaneous saturations by cryoprotectants.

BACKGROUND: The aim of this research was to study the effectiveness of perfusion of intact ovine ovaries with different rates of perfusion and time-period elapsed between extraction of these ovaries and the beginning of perfusion. METHODS: Ovaries were perfused through the arteria ovarica (ovarian arteries) with culture medium supplemented with 5% bovine calf serum, 6% dimethyl sulfoxide, 6% ethylene glycol, 0.15M sucrose, Indian ink, and 100 IU/mL heparin at room temperature (22 degrees C). In the first cycle of experiments, ovaries (n = 96) were perfused for 60 minutes just after extraction of ovaries at the following rates of perfusion (mL/hour): 150, 100, 75, 50, 25, 12.5, and 6.3. In the second cycle of experiments, ovaries (n = 26) were perfused at a rate of 25 mL/hour for 60 minutes after extraction of ovaries and their storage at room temperature for 2, 3, 4, and 5 hours, for groups 1, 2, 3, and, 4, respectively. Successful perfusion of blood vessels was detected visible by a blue coloration of the ovarian tissues. RESULTS: The first cycle of experiments showed that the optimal perfusion rates were 50 mL/hour and 25 mL/hour. In the second cycle of experiments, good perfusion of ovaries was established at a perfusion rate of 25 mL/hour for the ovaries of groups when 2 and 3 hours had elapsed after extraction. CONCLUSIONS: Effective perfusion of ovine intact ovaries with vascular pedicle was established using freezing medium at room temperature at the rate of perfusion of 25 mL/hour or 50 mL/hour. The ovaries must be perfused not later than 3 hours after the death of animals.

[S3 guidelines on diagnostics and treatment of cervical cancer: Demands on pathology]. / S3-Leitlinie Diagnostik und Therapie des Zervixkarzinoms.

Between 2011 and the end of 2014 the former consensus S2k guidelines for the diagnostics and treatment of cervical cancer were updated and upgraded to S3 level, methodologically based on the regulations of the German Cancer Society (DKG). The present article summarizes the relevant aspects for the sectioning, histopathological workup, diagnostics and reporting for the pathology of invasive cancer of the uterine cervix. The recommendations are based on the most recent World Health Organization (WHO) and TNM classification systems and consider the needs of the clinician for appropriate surgical and radiotherapeutic treatment of patients. Detailed processing rules of colposcopy-guided diagnostic biopsies, conization and trachelectomy as well as for radical hysterectomy specimens and lymph node resection (including sentinel lymph node resection) are given. In the guidelines deep stromal invasion in macroinvasive cervical cancer is defined for the first time as tumor infiltration of > 66% of the cervical stromal wall. Furthermore, morphological prognostic factors for microinvasive and macroinvasive cervical cancer are summarized.

Quality of human spermatozoa: relationship between high-magnification sperm morphology and DNA integrity.

The aim of this work is to establish the relationship between the morphology of Intracytoplasmic Morphologically Selected Sperm Injection (IMSI)-selected spermatozoa and their DNA integrity. The 45 ejaculates were randomly distributed into three treatment groups: normozoospermic, oligoasthenozoospermic and oligoasthenotheratozoospermic samples. The evaluation of DNA integrity was performed using the sperm chromatin dispersion test. It was established that DNA integrity of spermatozoa is strongly dependent on ejaculate quality (P < 0.05). The count of spermatozoa with nonfragmented DNA in normozoospermic samples was high and independent from IMSI-morphological classes (Class 1 versus Class 3, respectively) (P > 0.1). With decreased ejaculate quality, the percentage of spermatozoa with nonfragmented DNA decreased significantly (P < 0.05) independent from morphological class. Nevertheless, the rate of IMSI-selected spermatozoa with fragmented DNA within of Class 1 in normozoospermic (Group 1), in oligoasthenozoospermic (Group 2) and in oligoasthenotheratozoospermic (Group 3) samples was 21.1%, 31.8% and 54.1%, respectively. In conclusion, there is a direct relationship between morphological parameters of spermatozoa and their DNA integrity. However, the IMSI technique alone is not enough for the selection of spermatozoa with intact nuclei.

In vitro perfusion of whole bovine ovaries by freezing medium: effect of perfusion rate and elapsed time after extraction.

BACKGROUND: Cryopreservation and transplantation of the whole ovary with vascular pedicle would be helpful to prevent posttransplantation ischemia. In fact, perfusion of the intact mammalian ovary through arteries and veins is the most technically difficult part of the whole cryopreservation process because of its complexity. It is important to develop the technology of long-time perfusion of intact ovaries by cryoprotectants at low temperatures because it was established earlier that 24-hour cooling to 5 degrees C before cryopreservation is beneficial for the freezing of human ovarian tissue. The aim of this research was to study the effectiveness of perfusion of intact bovine ovaries with different rates of perfusion and elapsed time between extraction of these ovaries and beginning of perfusion. METHODS: Arteria ovarica was cannulated and ovaries were perfused with Leibovitz L-15 medium + 100 IU/mL heparin + 5% bovine calf serum + 6% dimethyl sulfoxide + 6% ethylene glycol + 0.15 M sucrose + Indian ink at room temperature (22 degrees C). In the first cycle of experiments, ovaries (n = 145) were perfused for 60 minutes during 1 to 1.5 hours after extraction of ovaries in the slaughter house at perfusion rates of 150 mL/hour (2.5 mL/minute), 100 mL/hour (1.67 mL/minute), 75 mL/hour (1.25 mL/minute), 50 mL/hour (0.83 mL/minute), 25 mL/hour (0.42 mL/minute), and 12.5 mL/hour (0.21 mL/minute) for groups 1, 2, 3, 4, 5, and 6, respectively. In the second cycle of experiments, ovaries (n = 29) were perfused with a rate of 25 mL/hour (0.42 mL/minute) for 60 minutes during the following time-periods elapsed after extraction of ovaries in the slaughter house: 3 hours (n = 18), 4 hours (n = 5), 5 hours (n = 3), and 6 hours (n = 3) for groups 1, 2, 3, and 4, respectively. Ovaries in luteal and follicular phase of development were distributed randomly into groups. Successful perfusion of blood vessels was detected visibly by a blue coloration of the vascular pedicle and ovarian tissues. The percentage of Indian ink-perfused tissues was detected. The intensity of the vascular leakage and tissue damage was scored microscopically and noted as follows: lack of disruption (-), weak disruption (+), moderate disruption (++), and strong disruption RESULTS: The first cycle of experiments shows that an optimal perfusion rate was established for groups 4 and 5 (50 and 25 mL/hour, respectively). In the second cycle of experiments, good perfusion of ovaries with the perfusion rate of 25 mL/hour was established only for ovaries of group 1 (3 hours after extraction). The effectiveness of perfusion in group 2 (4 hours after extraction) was sharply decreased. CONCLUSIONS: Effective perfusion of bovine intact ovaries with vascular pedicle with freezing medium (6% ethylene glycol + 6% dimethyl sulfoxide + 0.15 M sucrose) at room temperature includes a rate of perfusion 25 or 50 mL/ hour. Ovaries must be perfused no later than 3 hours after the death of animals.

Long-time cooling of human ovarian tissue before cryopreservation as obvious procedure: stimulation of follicular development and neo-vascularisation.

BACKGROUND: The positive effect of cooling on tissue cells is known. The aim of this research was to study the intensiveness of neo-vascularisation and follicular development in ovarian tissue after 24 hours cooling to 5 degrees C before cryopreservation. METHODS: Fifty six pieces from 7 patients were divided into the following four groups: Group 1: pieces cultured just after operation, Group 2: pieces cooled after operation to 5 degrees C for 24 hours and then cultured, Group 3: pieces frozen-thawed just after operation and then cultured, Group 4: pieces cooled after operation to 5 degrees C for 24 hours, frozen, thawed, and then cultured. Culture of ovarian pieces was performed in a chorioallantoic membrane (CAM)-system for 5 days. The efficacy of the tissue cooling was evaluated by the development of follicles and intensiveness of neo-vascularisation (by Desmin). RESULTS: For Group 1, 2, 3, and 4, mean density of follicles per 1 mm3 was 10.1, 11.1, 9.8, and 12.0, respectively (P1-2, 3-4 < 0.05). For these groups 91%, 92%, 90%, and 90% preantral follicles were morphologically normal (P1-2, 3-4 > 0.1). The immunohistochemical analysis showed that the intensiveness of neo-vascularisation observed in ovarian tissue of Group 2 (pre-cooled before culture) and Group 4 (pre-cooled before cryopreservation) was drastically increased. CONCLUSIONS: The 24 hour cooling to 5 degrees C before cryopreservation is beneficial for cryopreservation of human ovarian tissue.

Cryoprotectant-free vitrification of fish (Oncorhynchus mykiss) spermatozoa: first report.

The aims of this investigation were to test a novel technology comprising cryoprotectant-free vitrification of the spermatozoa of rainbow trout and to study the ability of sucrose and components of seminal plasma to protect these cells from cryo-injuries. Spermatozoa were isolated and vitrified using three different media: Group 1: standard buffer for fish spermatozoa, Cortland(®) medium (CM, control); Group 2: CM + 1% BSA + 40% seminal plasma; and Group 3: CM + 1% BSA + 40% seminal plasma + 0.125 m sucrose. For cooling, 20-µl suspensions of cells from each group were dropped directly into liquid nitrogen. For warming, the spheres containing the cells were quickly submerged in CM + 1% BSA at 37 °C with gentle agitation. The quality of spermatozoa before and after vitrification was analysed by the evaluation of motility and cytoplasmic membrane integrity with SYBR-14/propidium iodide staining technique. Motility (86%, 81% and 82% for groups 1, 2 and 3, respectively) (P > 0.1) was not decreased significantly. At the same time, cytoplasmic membrane integrity of spermatozoa of Groups 1, 2 and 3 was changed significantly (30%, 87% and 76% respectively) (P < 0.05). All tested solutions can be used for vitrification of fish spermatozoa with good post-warming motility. However, cytoplasmic membrane integrity was maximal in Group 2 (CM + 1% BSA + 40% seminal plasma). In conclusion, this is the first report about successful cryoprotectant-free cryopreservation of fish spermatozoa by direct plunging into liquid nitrogen (vitrification). Vitrification of fish spermatozoa without permeable cryoprotectants is a prospective direction for investigations: these cells can be successfully vitrified with 1% BSA + 40% seminal plasma.

Cryoprotectant-free vitrification of human spermatozoa in large (up to 0.5 mL) volume: a novel technology.

BACKGROUND: The aim of this study was to develop and to test the aseptic technology of cryoprotectant-free vitrification of human spermatozoa in large volume (for intrauterine insemination). Spermatozoa, vitrified by this technology, are free of permeant cryoprotectants and are ready for further use immediately after warming without any additional treatment (centrifugation or separation in the gradient for removal of cryoprotectant). METHODS: Each of 52 swim up-prepared ejaculates were divided into three aliquots and distributed into three treatment groups: Group 1: non-treated control; Group 2: spermatozoa cryopreserved by slow conventional freezing with glycerol-containing medium, and Group 3: spermatozoa vitrified in 0.5 mL insemination "French" straws in culture medium with 0.25 M sucrose. Sperm motility 1, 24 and 48 hours after warming, plasma membrane integrity and capacitation-like changes (spontaneous "cryo-capacitation" and acrosome reaction) were assessed after freezing-thawing. RESULTS: In contrast to conventional freezing, spermatozoa vitrified with aseptic cryoprotectant-free technology displayed superior functional characteristics. The motility rate, integrity rates of cytoplasmic, and acrosomal membranes were significantly higher after vitrification than after conventional freezing (76% vs 52%, 54% vs 28% and 44% vs 30%, respectively) (p < 0.05). However, there were no differences between vitrification and conventional freezing in the presence of glycerol in terms of percentage of spermatozoa expressing CTC-capacitation pattern (11% vs 10%, respectively) (p > 0.1). CONCLUSIONS: A basic protection from cryo-injury can be achieved for human spermatozoa using the novel technology of aseptic cryoprotectant-free vitrification in large volumes.

Serological and immunohistochemical HER-2/neu statuses do not correlate and lack prognostic value for ovarian cancer patients.

The serodiagnostics of extracellular domain (ECD) HER-2/neu has turned into an evidenced-based tumour marker for HER-2/neu-positive breast cancer patients. This study investigated the clinical relevance of immunohistochemical and serum HER-2/neu in 44 patients with advanced ovarian cancer. The Hercept-Test® from DAKO Diagnostics was used to analyse immunohistochemical HER-2/neu expression. The HER-2/neu ECD in serum was determined quantitatively by Bayer Immuno 1™ Immunoanalyser. The HER-2/neu serum values were correlated to the clinical course of disease and to established prognostic factors, i.e. progression-free and overall survival. Some 23% of patients (n = 11) expressed HER-2/neu serum levels higher than 15 ng/mL, whereas only 7.7% (n = 2) of the patients examined by immunohistochemistry showed a HER-2/neu overexpression of the tissue. None of them revealed an overexpression of HER-2/neu ECD by serodiagnostics. HER-2/neu overexpression did not correlate significantly to any of the analysed prognostic factors. According to progression-free and overall survival, there was no significant difference between serologically HER-2/neu-positive or negative patients. For ovarian cancer patients, neither high HER-2/neu serum levels, nor immunohistochemically determined HER-2/neu positivity, appear to predict the course of disease. This study shows a lack of association between the immunohistochemical HER-2/neu status and the serum level of solute extracelluar HER-2/neu domain.

Apoptosis in human ovarian tissue after conventional freezing or vitrification and xenotransplantation.

One of the new emerging techniques to preserve reproductive potential of cancer patients is cryopreservation of ovarian fragments prior to medical treatment and their retransplantation after healing. In order to investigate and compare apoptosis in human ovarian tissue after conventional ("slow") freezing and vitrification, we used a xenograft model in which conventionally frozen, vitrified and fresh non treated human ovarian tissue pieces were subcutaneously transplanted in SCID mice. The tissue samples were weekly, during four weeks, recovered from scarified SCID mice. The apoptosis was examined by immunohistochemical staining with the anti-caspase-3 antibody. There was a significant difference between the amount of apoptotic cells in cryopreserved ovarian tissue independent from mode of cooling compare to the control. The ovarian tissue after vitrification showed a significantly higher amount of apoptotic cells, than in slow frozen. The results obtained after comparative study of two different cryopreservation methods show that vitrification of human ovarian tissue could become a practice-relevant alternative to slow cryopreservation only after further improvement.

Relevance of anti-Mullerian hormone measurement in a routine IVF program.

BACKGROUND: Diminished ovarian reserve has become a major cause of infertility. Anti-Mullerian hormone (AMH) seems to be a promising candidate to assess ovarian reserve and predict the response to controlled ovarian hyperstimulation (COH). This prospective study was conducted to evaluate the relevance of AMH in a routine IVF program. METHODS: Three hundred and sixteen patients were prospectively enrolled to enter their first IVF/ICSI-cycle. Age, FSH-, inhibin B- and AMH-levels and their predictive values for ovarian response and clinical pregnancy rate were compared by discriminant analyses. RESULTS: A total of 132 oocyte retrievals were performed. A calculated cut-off level < or =1.26 ng/ml AMH alone detected poor responders (< or =4 oocytes) with a sensitivity of 97%, and there was a 98% correct prediction of normal response in COH if levels were above this threshold. With levels <0.5 ng/ml, a correct prediction of very poor response (< or =2 oocytes) was possible in 88% of cases. Levels of AMH > or =0.5 ng/ml were not significantly correlated with clinical pregnancy rates. CONCLUSIONS: AMH is a predictor of ovarian response and suitable for screening. Levels < or =1.26 ng/ml are highly predictive of reduced ovarian reserve and should be confirmed by a second line antral follicle count. Measurement of AMH supports clinical decisions, but alone it is not a suitable predictor of IVF success.

"Occult" neuroendocrine component and rare metastatic pattern in cervical cancer: report of a case and brief review of the literature.

Distant metastases in small cell carcinomas of the uterine cervix are rare, and a disseminated manifestation of the disease is uncommon. This is a case report of a 40-year-old woman treated with platin-based radio-chemotherapy for a moderately differentiated squamous cell cervical cancer FIGO Stage IB 1 (with positive paraaortic lymph nodes). One year later she presented with remarkably unusual cutaneous metastases of the left thumb and scalp as the first signs of spread of disease, including kidney, lung and brain metastases. An advanced retrospective immunohistochemical staining of the cervical biopsy discovered a small neuroendocrine component of the carcinoma as the presumably causative factor for the rare metastastic pattern and poor prognosis.

Technologies of cryoprotectant-free vitrification of human spermatozoa: asepticity as criterion of effectiveness.

This review describes 120 years history of technology for cryoprotectant-free cryopreservation of human spermatozoa by direct plunging into liquid nitrogen (vitrification). It is presented an explanation why cryoprotectant-free vitrification for some human ejaculates is better than conventional freezing and vitrification with the presence of cryoprotectants. Special attention is given to the extremely high viability of viruses, bacteria and micoplasmas after cryoprotectant-free cryopreservation in culture medium and even in distilled water. This fact increases the potential risk of disease transmission through liquid nitrogen. It is concretized the concept "asepticity" as obvious parameter for any medical assisted reproduction technology which includes the cooling of cells in liquid nitrogen. It is described the role of nonpermeable compounds of mediums for cryoprotectant-free vitrification: carbohydrates, proteins, lipoproteins, antioxidants. This review summarizes concerned data regarding two groups of different current technologies for cryoprotectant-free vitrification of human spermatozoa: with direct contact of spermatozoa with liquid nitrogen as well as with full isolation of these cells from liquid nitrogen (aseptic technologies).

New Screening Proposals: the Federal Joint Commission Defines the Parameters for Cervical Cancer Screening from 2018: Statement of the Gynecology Oncology Working Group (AGO).

The Gynecology Oncology Working Group (AGO e. V.) unequivocally welcomes the decision taken by the German Federal Joint Commission (Gemeinsamer Bundesausschuss, G-BA) on March 19, 2015 regarding screening for cervical cancer. AGO is convinced that, in view of recent medical advances, this evidence-based decision will improve screening for cervical cancer.

Induction of Labour in Late and Postterm Pregnancies and its Impact on Maternal and Neonatal Outcome.

INTRODUCTION: This study aimed to determine the effects of induction of labour in late-term pregnancies on the mode of delivery, maternal and neonatal outcome. METHODS: We retrospectively analyzed deliveries between 2000 and 2014 at the University Hospital of Cologne. Women with a pregnancy aged between 41 + 0 to 42 + 6 weeks were included. Those who underwent induction of labour were compared with women who were expectantly managed. Maternal and neonatal outcomes were evaluated. RESULTS: 856 patients were included into the study. The rate of cesarean deliveries was significantly higher for the induction of labour group (33.8 vs. 21.1 %, p < 0.001). Aside from the more frequent occurrence of perineal lacerations (induction of labour group vs. expectantly managed group = 38.1 % compared with 26.4 %, p = 0.002) and all types of lacerations (induction of labour group vs. expectantly managed group = 61.5% vs. 52.2 %, p = 0.021) in women with vaginal delivery, there were no significant differences in maternal outcome. Besides, no differences regarding neonatal outcome were observed. CONCLUSIONS: Our study suggests that induction of labour in late and postterm pregnancies is associated with a significantly higher cesarean section rate. Other maternal and fetal parameters were not influenced by induction of labour.

Diagnosis, Therapy and Follow-up Care of Vulvar Cancer and its Precursors. Guideline of the DGGG and DKG (S2k-Level, AWMF Registry Number 015/059, November 2015.

Purpose: This is an official guideline, published and coordinated by the Arbeitsgemeinschaft Gynäkologische Onkologie (AGO, Study Group for Gynecologic Oncology) of the Deutsche Krebsgesellschaft (DKG, German Cancer Society) and the Deutsche Gesellschaft für Gynäkologie und Geburtshilfe (DGGG, German Society for Gynecology and Obstetrics). The number of cases with vulvar cancer is on the rise, but because of the former rarity of this condition and the resulting lack of literature with a high level of evidence, in many areas knowledge of the optimal clinical management still lags behind what would be required. This updated guideline aims to disseminate the most recent recommendations, which are much clearer and more individualized, and is intended to create a basis for the assessment and improvement of quality care in hospitals. Methods: This S2k guideline was drafted by members of the AGO Committee on Vulvar and Vaginal Tumors; it was developed and formally completed in accordance with the structured consensus process of the Association of Scientific Medical Societies in Germany (Arbeitsgemeinschaft der Wissenschaftlichen Medizinischen Fachgesellschaften, AWMF). Recommendations: 1. The incidence of disease must be taken into consideration. 2. The diagnostic pathway, which is determined by the initial findings, must be followed. 3. The clinical and therapeutic management of vulvar cancer must be done on an individual basis and depends on the stage of disease. 4. The indications for sentinel lymph node biopsy must be evaluated very carefully. 5. Follow-up and treatment for recurrence must be adapted to the individual case.

Strong evidence that the common variant S384F in BRCA2 has no pathogenic relevance in hereditary breast cancer.

INTRODUCTION: Unclassified variants (UVs) of unknown clinical significance are frequently detected in the BRCA2 gene. In this study, we have investigated the potential pathogenic relevance of the recurrent UV S384F (BRCA2, exon 10). METHODS: For co-segregation, four women from a large kindred (BN326) suffering from breast cancer were analysed. Moreover, paraffin-embedded tumours from two patients were analysed for loss of heterozygosity. Co-occurrence of the variant with a deleterious mutation was further determined in a large data set of 43,029 index cases. Nature and position of the UV and conservation among species were evaluated. RESULTS: We identified the unclassified variant S384F in three of the four breast cancer patients (the three were diagnosed at 41, 43 and 57 years of age). One woman with bilateral breast cancer (diagnosed at ages 32 and 50) did not carry the variant. Both tumours were heterozygous for the S384F variant, so loss of the wild-type allele could be excluded. Ser384 is not located in a region of functional importance and cross-species sequence comparison revealed incomplete conservation in the human, dog, rodent and chicken BRCA2 homologues. Overall, the variant was detected in 116 patients, five of which co-occurred with different deleterious mutations. The combined likelihood ratio of co-occurrence, co-segregation and loss of heterozygosity revealed a value of 1.4 x 10-8 in favour of neutrality of the variant. CONCLUSION: Our data provide conclusive evidence that the S384F variant is not a disease causing mutation.

Mitoxantrone combined with paclitaxel as salvage therapy for platinum-refractory ovarian cancer: laboratory study and clinical pilot trial.

This report describes preclinical and early clinical investigations of the mitoxantrone/paclitaxel combination (NT) for patients with platinum-refractory ovarian cancer. The preclinical activity of NT was studied ex vivo, evaluating native tumor specimens with the ATP tumor chemosensitivity assay. Of 24 tumors tested, 20 (83%) were sensitive to NT, whereas 7 (29%) responded to mitoxantrone and 8 (33%) responded to paclitaxel. In the majority of tumors assayed (19 of 24), potentiating or major independent effects between both agents were found. Subsequently, a clinical pilot trial of NT was initiated for patients with platinum-refractory ovarian cancer. Patients had failed one to four (median, two) prior chemotherapy regimens. In 11 cases, NT was administered every three weeks with 8 mg/m2 mito-xantrone and 180 mg/m2 paclitaxel (NT-I). Seven patients were treated biweekly with 6 mg/m2 mitoxantrone and weekly with 100 mg/m2 paclitaxel (NT-II). During 92 NT courses, myelosuppression with leucopenia, anemia, and thrombocytopenia was the limiting toxicity, occurring more frequently with NT-II. No patient required hospitalization due to any life-threatening complication. Five complete and nine partial remissions were observed with both NT-I and NT-II, accounting for an overall 78% response rate, with a median progression-free survival of 40 weeks. One patient showed early progression during therapy. Currently, three patients (NT-I, two; NT-II, one) have died due to progressive relapsed ovarian cancer, so that the median overall survival is not reached after a median follow-up of 40.5+ weeks. Both schedules were found to be equal in terms of response rate and overall survival. NT is highly active and practical for salvage treatment of ovarian cancer. NT-II may be preferred due to both clinical activity and patients' acceptance. However, NT-I seems to be a less myelotoxic alternative. Both schedules warrant further clinical investigation.