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Objective This study aimed to validate non-invasive RHD genotyping of cell-free fetal DNA (cff-DNA) using different DNA extraction methods and of fresh and frozen extracted cff-DNA. Background Non-invasive RHD genotyping of cff-DNA predicts fetal RhD phenotype, allowing for the rational implementation of antenatal immunoprophylaxis and representing a big step forward in the management of RhD-immunised women. Validation of a diagnostic method is mandatory before its clinical application. Methods RhD-negative pregnant women were recruited at different gestational ages. The cff-DNA extraction was carried out using manual and automatic methods in order to improve cff-DNA yield and optimise the extraction. Fetal RHD genotyping was performed using a commercial real-time polymerase chain reaction (PCR) kit, and the results were compared with postnatal serological RhD determination on cord blood. Results Overall, 133 plasma samples were examined for the validation process, and a total of 423 tests were performed. No differences have been observed between the two extraction methods or between fresh or frozen cff-DNA regarding cff-DNA stability and quality parameters. There was 100% concordance between fetal RHD genotyping of cff-DNA and RhD phenotype on cord blood for both extraction methods on both fresh and frozen cff-DNA. Conclusion Our study shows the reliability of automatic and manual cff-DNA extraction methods and the possibility of freezing extracted cff-DNA when performing RHD genotyping. This result might be relevant for improving laboratory work and organisation through the development of a standardised procedure for fetal RHD genotyping on cff-DNA, laying the foundations for evidence-based use of anti-D Ig prophylaxis in RhD pregnant women.

Stereotactic body radiotherapy (SBRT) is a new treatment modality for prostate cancer. The current study evaluates CyberKnife® SBRT and reports toxicity and early Prostate-Specific Antigen (PSA) kinetics. From June 2006 to August 2009, 45 low-and intermediate-risk prostate cancer patients received Cyberknife SBRT of 35 Gy in five fractions with 95% minimum target coverage. Median follow-up was 20-months (range 6–42-months). Seventeen patients received androgen-deprivation therapy also. Acute complications were mild, short-lived and no greater than Grade 2 by RTOG scale. Late toxicities consisted of one patient (2.2%) experiencing Grade 2 rectal, one patient (2.2%) Grade 3 and four patients (8.8%) with Grade 1 urinary toxicity. PSA in all patients progressively declined from a mean 4.7 ng/ml baseline to 1.48 ng/ml at three months, to 0.68 ng/ml at 12 months and to 0, 35 ng/ml at 24 months. The 28 hormon-naive patients had the mean PSA value of 1.1 ng/ml at one year from a mean 6.65 ng/ml baseline. There was a significant PSA value reduction in 11 hormone therapy patients with low baseline PSA value (< 1 ng/ml) from 0.37 down 0.14 ng/ml (p value 0.0068) at one year. Moreover, 14 low risk patients gave better results of mean PSA value than 17 Intermediate risk patients 0.43 ng/ml vs. 0.93 ng/ml (p value 0.02) at one year. No patient had biochemical failure at last follow-up. Hypofractionated SBRT appears to have potential against prostate cancer. Low toxicity and encouraging biochemical control support its use in early-stage prostate cancer. Results encourage further follow-up and larger studies.

Carbohydrates play an important role in the stabilization of biological components under water stress and, consequently, in the preservation of foods. Water stress is produced either by freezing or drying producing damage in proteins and tissues mainly due to the generation of large gradients of concentration of salts. Some carbohydrates act as protectors but, although some explanations have been proposed, the mechanism is not yet properly known. We have measured the dielectric relaxation times of aqueous solutions of glucose in the time domain with a custom made device. Some results are presented here for a solution of glucose in water at 88% w/w, in the range of the glass transition temperature, including some data on the rubbery state (above T g ). The curves were fitted with a stretched exponential decay (KWW) with relaxation times in the range of 200–500s and stretching parameter ( β ) around 0.5.

Hydraulic head response to stream-stage variations can be used to explore the hydraulic properties of stream-aquifer systems at a relatively large scale. These stream-stage response tests, also called flooding tests, are typically interpreted using one- or two-dimensional models that assume flow perpendicular to the river. Therefore, they cannot be applied to systems that are both horizontally and vertically heterogeneous. In this work, we use the geostatistical inverse problem to jointly interpret data from stream-stage response and pumping tests. The latter tests provide flow data (which are needed to resolve aquifer diffusivity into transmissivity and storage coefficient) and may supply supplementary small-scale information. Here, we summarize the methodology for the design, execution, and joint numerical interpretation of these tests. Application to the Aznalcóllar case study allows us to demonstrate that the proposed methodology may help in responding to questions such as the continuity of aquitards, the role and continuity of highly permeable paleochannels, or the time evolution of stream-aquifer interaction. These results expand the applicability and scope of stream-stage response tests.

Purpose: The combination of irradiation and total mesorectal excision for rectal carcinoma has significantly lowered the incidence of local recurrence. However, a new problem is represented by the patient with locally recurrent cancer who has received previous irradiation to the pelvis. In these patients, local recurrence is very often not easily resectable and reirradiation is expected to be associated with a high risk of late toxicity. The aim of this multicenter phase II study is to evaluate the response rate, resectability rate, local control, and treatment-related toxicity of preoperative hyperfractionated chemoradiation for locally recurrent rectal cancer in patients previously irradiated to the pelvis. Methods and Materials: Patients with histologically proven pelvic recurrence of rectal carcinoma, with the absence of extrapelvic disease or bony involvement and previous pelvic irradiation with doses ≤55 Gy; age ≥18 years; performance status (PS) (Karnofsky) ≥60, and who gave institutional review board–approved written informed consent were treated by preoperative chemoradiation. Radiotherapy was delivered to a planning target volume (PTV2) including the gross tumor volume (GTV) plus a 4-cm margin, with a dose of 30 Gy (1.2 Gy twice daily with a minimum 6-h interval). A boost was delivered, with the same fractionation schedule, to a PTV1 including the GTV plus a 2-cm margin (10.8 Gy). During the radiation treatment, concurrent chemotherapy was delivered (5-fluorouracil, protracted intravenous infusion, 225 mg/m 2 /day, 7 days per week). Four to 6 weeks after the end of chemoradiation, patients were evaluated for tumor resectability, and, when feasible, surgical resection of recurrence was performed between 6–8 weeks from the end of chemoradiation. Adjuvant chemotherapy was prescribed to all patients, using Raltitrexed, 3 mg/square meter (sm), every 3 weeks, for a total of 5 cycles. Patients were staged using the computed tomography (CT)-based F-classification (F0: no side-wall involvement; F1, F2, F3: 1, 2, and 3–4 side-walls involved, respectively). Toxicity was evaluated on the basis of the Radiation Therapy Oncology Group (RTOG) criteria. Results: Fifty-nine patients (38 male, 21 female; median age, 62 years; range, 43–77 years) were enrolled in the study, by 12 different Italian radiotherapy departments. Previous surgery was anterior resection in 45 patients (76.3%) and abdominal-perineal resection in 14 patients (23.7%); previous radiotherapy dosage ranged between 30 and 55 Gy (median, 50.4 Gy); the median interval between prior radiation therapy to the onset of reirradiation was 27 months (range, 9–106 months); 44 patients (74.6%) had received some form of previous chemotherapy (concurrent and/or adjuvant). Fifty-one of 59 patients (86.4%) completed chemoradiation without treatment interruptions: 6 patients (10.2%) had temporary treatment interruption due to toxicity or patient compliance, and 2 patients (3.4%) had definitive treatment interruption. The incidence of Grade 3 lower gastrointestinal acute toxicity was only 5.1%. No patient developed Grade 4 acute toxicity. After chemoradiation, 5 patients (8.5%) had complete response (CR), 21 patients (35.6%) had partial response (PR), 31 patients (52.6%) had no change (NC) and 2 patients (3.4%) showed progressive disease (PD). Overall, the response rate (PR + CR) was 44.1% (95% confidence interval, 29.0–58.9%). Twenty of 24 patients (83.3%) with pelvic pain before treatment had symptomatic response. Tumor resection was performed in 30 of 59 patients (50.8%) including 2 local excisions, 4 anterior resections, 18 abdominoperineal resections, and 6 other. Surgical resection resulted as R0 and R1 in 21 patients (35.6%) and 3 patients (5.1%), respectively. The possibility of radical resection was influenced by tumor response to chemoradiation (PD/NC: 7/33; PR/CR: 14/26; p = 0.009). Thirty-three patients received adjuvant chemotherapy, which was completed in 30 (50.8%). At a median follow-up of 36 months (range, 9–69 months), 28 patients (47.5%) developed local recurrence or tumor progression in the unresected pelvic disease and 18 patients (30.5%) developed distant metastasis. Seven patients showed late toxicity, including 2 skin fibrosis, 2 impotence, 2 urinary complications requiring nephrostomy, and 1 small bowel fistula requiring surgical diversion. Overall median survival was 42 months. Five-year actuarial survival was 39.3%; 66.8% in R0 resected patients and 22.3% in patients treated without surgery or undergoing subtotal tumor removal. Local control and disease-free survival were significantly correlated with the interval between surgical treatment for primary tumor and local recurrence ( p = 0.028 and p = 0.003, respectively). Radical resection significantly influenced local control, disease-free survival, and overall survival ( p = 0.010, p = 0.010, and p = 0.050 respectively). The multivariate analysis confirmed the impact of surgery-relapse interval on local control ( p = 0.016) and disease-free survival ( p = 0.002), and confirmed the correlation between R0 surgery with local control and disease-free survival ( p = 0.016). Conclusions: Use of hyperfractionated chemoradiation was associated with a low rate of acute toxicity and an acceptable incidence of late complications. Pain control was excellent. The overall 5-year survival was 39%. Despite 87.4% of patients having F1–3 stage disease, approximately one-third (35%) achieved R0 resection, and two-thirds of patients in this cohort of patients were alive at the 5-year mark. However, further studies using innovative treatment algorithms are warranted to, hopefully, improve the local tumor response and control.

The alluvial deposits of the Agrio River in SW Spain have been studied using terrace mapping, boreholes, trenching and vertical electrical sounding to select an adequate place for a Permeable Reactive Barrier. Geological and hydrogeological data available prior to the barrier construction suggested a simple geological model based on three terraces, the most modern being deeper than the oldest. The barrier was accordingly trenched through the youngest terrace. However, excavation of the barrier andsubsequent subsoil data demonstrated that the internal structure of the Agrio alluvial deposits does not follow such a simple model. A revised model, less favourable to the existing barrier design, revealed the oldest terrace to be deeper than the youngest, and to form a palaeochannel oblique to the surface terraces and river trends. The study methods used are standard, but proved to have insufficient resolution to detect key features of the alluvial geology. It is concluded that such characterization methods, though widely used, are not appropriate where alluvial terraces may display complex internal structures not reflected in the modern surface geomorphology. In hindsight, the study area should have been larger, so as to encompass at least the width of the alluvial plain and to extend for a hundred metres or so up and downstream from the proposed barrier location. Lithological logs of the boreholes should have been carefully described from both cuttings and downhole geophysical logs, which would have allowed more accurate delineation ofthe stratigraphy of the alluvial deposit. Subsequentgeophysical methods should have been calibrated to this stratigraphy to characterize the internal structure and basal contact of the alluvial deposit.

Static permittivity and dielectric relaxation measurements in microemulsions-based gels (organogels) were performed. Dielectric spectra have been obtained with a time domain spectrometer in the range 100KHz to 1 GHz. These systems were composed of AOT/ water/iso-octane and gelatin and microemulsions were prepared by changing W 0 = [AOT/water] and the AOT molarity (M). Experimental results of dielectric parameters as a function of gelatin concentration are presented. It must be pointed out that unlike other techniques no clear percolative behaviour can be deduced from this kind of experimental technique. Assuming an interfacial polarization mechanism, we were able to calculate the basic electrical length regarding the polarization. These results have been analysed by comparing them with those which have been obtained with other experimental techniques.

A retrospective analysis is reported in 232 patients with clinical Stage I endometrial adenocarcinoma diagnosed between 1980 and 1988, treated with combined surgery and adjuvant irradiation. Tumors were restaged according to the FIGO (1988) surgical staging system (SSS) in order to assess the prognostic value of this revised classification and of the conventional clinico-pathological features (age, performance status, grade and depth of myometrial invasion). The 5-year relapse-free survival (RFS) and overall survival (OS) of the series was 85% and 90.8%, respectively. Both 5-year RFS and OS were not significantly different adopting the FIGO (1971) clinical Stage (Stage IA 93% and 88% vs Stage IB 88.5 and 82%. respectively) whereas they were significant by FIGO SSS (Stage IA 96% and 93% vs Stage IB 94% and 88.5% vs Stage IC 74% and 63%, respectively) (p=0.001 and p=0.0005, respectively). Other factors that significantly affected 5-year RFS or OS at univariate analysis were age (p=0.01 and p

Thirty women with locally advanced breast cancer (LABC), but no evidence of distant metastases, were prospectively treated with four fixed cycles of neoadjuvant chemotherapy (CT). This regimen consisted of epidoxorubicin (Epi) alternated every 21 days with cyclophosphamide, methotrexate and 5-fluorouracil (CMF). After this induction CT, subsequent therapy was planned according to the response obtained as follows: (a) modified mastectomy with axillary dissection was performed in patients who had major objective response (complete or partial), followed by four doses of adjuvant CT and radiotherapy (RT); (b) debulking rescue surgery followed by RT and 2nd line CT with mitomycin C were given in patients with stable disease or minor response. The response rate to induction CT was 63% (19 of 30 patients) (95% confidence limits 46-80%). Overall, 43% of patients had no persistance of tumor at the end of the planned therapy. After a median follow-up time of 36 months, disease-free survival (DFS) and overall survival (OS) were 35% and 47%, respectively. The median duration of DFS was 16 + months (4-52+ months). A significantly better OS was observed in complete responders compared to the others (77% versus 23.5%; p=0.01). Compliance to treatment was high, gastrointestinal and hematological toxicities were the most common side-effects. Thus, this multimodal approach is effective in reducing primary tumor size with acceptable morbidity. Five of the 11 (45%) patients non responsive to induction CT obtained a transient local control of disease after debulking surgery, RT and mitomycin C. To assess the role of alternating non cross resistant regimens as induction therapy in LABC vs conventional schedules, phase III comparative studies are warrented.

Primary soft tissue sarcoma of the retroperitoneum is a rare disease. A series of 11 evaluable adult patients with retroperitoneal soft tissue sarcomas is reported. These patients were treated with complete surgery and adjuvant radiation therapy (total dose from 50 to 64 Gy) using an 18 MeV linear accelerator. After a median follow-up of 48 months (range, 6-84), 4 patients had a local-regional recurrence, 3 had distant metastases, and 4 died of progressive disease. Four-year estimated disease-free survival was 54.5% and overall survival was 70%. Treatment was well tolerated by most patients: 7 patients experienced moderate gastrointestinal toxicity, mainly nausea and diarrhea, during radiotherapy; 2 cases had weight loss > 15% at the end of the therapy; and chronic ileitis was observed in 2 cases. We conclude that adjuvant radiotherapy seems to reduce the incidence of local-regional recurrences in these patients. No radiation-induced irreversible injury was observed, but one young woman had amenorrhea after radiotherapy. Controlled clinical trials are warrented to define the role and effiectiveness of adjuvant radiotherapy and/or chemotherapy in retroperitoneal soft tissue sarcomas.

Between January 1987 and June 1991, 173 patients with inoperable non-small cell lung cancer, Stage III, were entered into a randomized trial comparing radiotherapy only (RT) (45 Gy/15 fractions/3 weeks) (arm A) versus RT and a daily low dose of cDDP (6 mg/m2) (arm B). An overall response rate of 58.9% was observed in arm A and 50.6% in arm B, respectively. No differences in the pattern of relapse were noted between the two treatment groups. Median time to progression was 10.6 months for arm A and 14.2 months for arm B. Median survivals were 10.3 months and 9.97 months, respectively. Toxicity was acceptable and no treatment-related death occurred in either treatment schedule. In this study no significant advantage of the combined treatment over radiation therapy only was found. The encouraging results achieved in some trials together with the intractability of the disease suggest that further efforts should be made to optimize clinical trial protocols, perhaps by reviewing the radiobiological and pharmacological basis of the combined treatment.

A multicenter retrospective study was undertaken in northern Italy to assess clinical characteristics and pattern of care of elderly patients (> 70 years) with lung cancer seen in 1984, at 14 non-surgical institutions. Overall, 1 out of 5 patients with lung cancer was elderly. Eighty-four percent of the 264 elderly patients under study were males, and overall, their median age was 76 years. Squamous cell carcinoma was the most frequent histologic type (53%), and most of the patients (61%), after a rather intensive diagnostic workup, had loco-regional disease. Sixty-three percent of patients underwent radiotherapy, which was mostly classified as palliative. Chemotherapy was given to 13% of patients, mostly with small cell histology. Response to radiotherapy and chemotherapy was observed in about 50% of the patients; however, 3 out of 34 (10%) patients treated with chemotherapy died from toxicity. The median survival time from diagnosis was 9.9 months; survival time was not related, in this population of elderly patients, to age. Throughout the study, the quality of data recording during the patient's illness was often found to be suboptimal. In conclusion, elderly patients with lung cancer appear to be actively staged and treated outside a protocol context in the clinical practice of cooperating nonsugical institutions in northern Italy. This stresses the importance of prospective studies aimed to evaluate the cost/effectiveness of the various procedures used and the impact of treatments used (or even of a policy of no treatment) on the survival and quality of life of these patients.

El riesgo hidrológico asociado a las crecidas de proyecto para el caso de cuencas pequeñas y medianas puede representarse a través de la probabilidad de excedencia del caudal pico, pero esto no ocurre en el caso de grandes cuencas con regímenes de crecidas complejos, dando lugar a que a una misma crecida real le corresponda un período de retorno diferente según se efectúe el análisis de frecuencia del caudal máxima, el volumen o la duración de la misma. Se estudian las relaciones que existen entre las variables mencionadas a partir de los caudales diarios disponibles en el tramo media del Río Paraná en la Argentina y se efectuan los análisis de frecuencia de extremos. En la zona de estudios las medidas estructurales y no estructurales de control y protección de crecidas se han proyectado en forma aislada, utilizando la característica de la crecida más apropiada para cada caso. Con el fin de verificar cual es el comportamiento de dichas obras existentes con crecidas de un mismo nivel de riesgo y adoptar la misma como crecida de proyecto de medidas complementarias, a partir de las relaciones estudiadas se definió un índice de crecida como el cociente entre el caudal máxima y el media de la crecida. Con este índice que integra las distintas variables en juego se efectuó el análisis de frecuencia y se estableció un procedimiento para adoptar valores de caudal máximo, volumen y duración que permiten la determinación de hidrogramas de diseño correspondientes a un determinado período de retorno.

From June 1991 to August 1994, 61 patients with stage III unresectable non-small-cell lung cancer (NSCLC; 16 cases of stage IIIA with N2 bulky disease and 45 cases of stage IIIB) were treated with ifosfamide given i.v. at 3 g/m2 on day 1, carboplatin given i.v. at 200 mg/m2 on days 1 and 2, etoposide given i.v. at 120 mg/m2 on days 1-3 (ICE) and recombinant human granulocyte colony-stimulating factor (rhG-CSF) given s.c. at 5 micrograms/kg on days 4-13. Chemotherapy was given every 3 weeks for up to three cycles and, unless the disease progressed, was followed by thoracic radiotherapy on the tumor volume (total dose 60 Gy) and mediastinum (40 Gy). All patients had measurable or evaluable unresectable disease and a performance status (Eastern Cooperative Oncology Group) of 0-1. Only 61% of the enrolled patients received the full program of chemoradiotherapy according to the study design. At the end of sequential chemo-radiotherapeutic treatment, 41% of the patients had an objective response (24 partial responses and 1 complete response), 31% showed no change and 28% had progressive disease. The response rate noted for patients in stage IIIA with N2 bulky disease and that recorded for patients in stage IIIB did not differ significantly. The median time to progression was 5.4 months and the median survival was 8.2 months, with the 1-year survival rate being 31%. Sites of progression were mostly intrathoracic. Haematological toxicity was the main side effect, with grade III-IV thrombocytopenia being reported in 24% of the 165 courses of intensive ICE chemotherapy given. Febrile neutropenia was described in six courses (three patients). Non-haematological toxicities and radiotherapy-related side effects were generally mild and easily manageable. In conclusion, in unresectable stage III NSCLC a short program of moderately intensified ICE chemotherapy with rhG-CSF protection followed by sequential radiotherapy failed to increase the percentage of objective responses and reached a median survival comparable with that previously achieved with standard doses.

9 Background: The purpose of this study is to report biochemical relapse-free survival (bRFS) rates for a group of localized prostate cancer patients from a pooled multi-institutional dataset with at least 5 years follow-up after stereotactic body radiotherapy (SBRT). Methods: The outcome data from 1101 patients treated with SBRT between 2003 and 2011 were pooled from 8 institutions. A subset of 135 cases had a minimum 5 years follow-up. All 135 cases had clinical stage T1 or T2A disease. The distribution by Gleason score (GS) was 38 Gy in 11%. Androgen deprivation therapy was given to 21% of patients. Biochemical relapse, defined as a rise > 2 ng/ml above nadir, was determined in a total of 4 failures. Results: The median follow-up for all 135 cases was 60 months (range 60 to 72). For all patients, the bRFS rate at 5 years was 97%. The 5-year actuarial bRFS rates for GS < 6, and Gleason score 7 were 98%, and 92%, respectively (p=0.15). The 5-year actuarial bRFS rates for low versus intermediate/high-risk patients were 99% and 93%, respectively (p=0.11). The 5-year actuarial bRFS rates for patients receiving 35 Gy versus >36.25 Gy were 93% and 100%. No difference in bRFS was observed with the use of androgen deprivation (p=0.78). Multivariate analysis showed only GS to be an independent predictor of relapse (p=0.03); iPSA (p=0.10) and radiation dose (0.97) were not. Conclusions: In a relatively large cohort of localized prostate cancer patients treated with SBRT, long follow-up period (>5 years), excellent efficacy was demonstrated with 97% of patients being free from relapse. For low and intermediate risk cases, these results compare favorably with other modalities with similar follow-up periods. Although a trend for worse outcome was seen with total radiation doses of 35 Gy, this was not confirmed on multivariate analysis.