Concordance of 3 alternative teratogenicity assays with results from corresponding in vivo embryo-fetal development studies: Final report from the International Consortium for Innovation and Quality in Pharmaceutical Development (IQ) DruSafe working group 2.

An IQ DruSafe working group evaluated the concordance of 3 alternative teratogenicity assays (rat whole embryo culture, rWEC; zebrafish embryo culture, ZEC; and murine embryonic stem cells, mESC) with findings from rat or rabbit embryo-fetal development (EFD) studies. Data for 90 individual compounds from 9 companies were entered into a database. In vivo findings were deemed positive if malformations or embryo-fetal lethality were reported in either species. Each company used their own criteria for deciding whether the alternative assay predicted the in vivo findings. Standard concordance parameters were calculated, positive and negative predictive values (PPV and NPV) were adjusted for the aggregate portfolio prevalence of positive compounds (established by a survey of participating companies), and positive and negative likelihood ratios (LR+ and iLR-) were calculated. Of the 3 assays, only rWEC data were robustly predictive, particularly for negative predictions (NPVadj = 92%). However, both LR+ (4.92) and iLR- (4.72) were statistically significant for the rWEC assay. When analyzed separately for rats, the NPVadj and iLR-values for the rWEC assay increased to 96% and 9.75, respectively. These data suggest that a negative rWEC outcome could defer or replace a rat EFD study in certain regulatory settings.

Gracilis and semitendinosus moment arm decreased by fascial tissue release after hamstring harvesting surgery: a key parameter to understand the peak torque obtained to a shallow angle of the knee.

PURPOSE: Semitendinosus and gracilis muscles whose tendons are used in surgical reconstruction of the anterior cruciate ligament maintain their contractile ability, but the peak torque angle of hamstring muscles shifted to a shallow angle postoperatively. The goal was to quantify the influence of the myofascial structures on instantaneous moment arms of knee muscles to attempt explaining the above-mentioned post-surgical observations. METHODS: Hamstring harvesting procedures were performed by a senior orthopaedic surgeon on seven lower limbs from fresh-frozen specimens. Femoro-tibial kinematics and tendons excursion were simultaneously recorded at each step of the surgery. RESULTS: No significant difference was demonstrated for instantaneous moment arm of gracilis during anterior cruciate ligament surgery (84% of the maximum intact values; P ≥ 0.05). The first significant semitendinosus moment arm decrease was observed after tendon harvesting (61% of the maximum intact values; p ≤ 0.005). After hamstring harvesting, the maximum and minimum moment arm (both gracilis and semi tendinosus) shifted to a shallow angle and 90°, respectively. CONCLUSION: Moment arm modifications by paratenons and the loose connective tissue release are essential to understand the peak torque obtained to a shallow angle. LEVEL OF EVIDENCE: Basic science study, biomechanics.

No impact of blood transfusion on oncological outcome after radical prostatectomy in patients with prostate cancer.

PURPOSE: To assess the association between blood loss, blood transfusion (BT) and biochemical recurrence (BCR)-free, metastasis-free and overall survival after radical prostatectomy (RP) in a large single-center cohort of patients. Perioperative BT at oncologic surgery has been reported to be a potential risk factor for cancer recurrence and survival in several cancer entities. Current studies addressing the relationship between BT, blood loss and BCR-free survival in prostate cancer patients are controversial and include only series with fairly small patient cohorts. MATERIALS AND METHODS: The data of 11,723 patients who underwent RP between 01/1992 and 08/2011 were analyzed. Cox regression analysis, including preoperative PSA level, pT stage, lymph node status, Gleason score, margin status, blood loss, transfusion rate (allogeneic or autologous), tested the relationship between blood loss, transfusion and BCR-free, metastasis-free and overall survival. Additionally, propensity score-matching analysis was performed to adjust differences in tumor characteristics. RESULTS: There was no statistically significant relationship between blood loss or BT and BCR-free, metastasis-free or overall survival. In multivariate analysis PSA level, pT stage, Gleason score, margin status and lymph node status were independent factors for a BCR (p < 0.0001). These results were identical after propensity score matching analysis, comparing patients with and without BT. CONCLUSIONS: This large-scale analysis revealed no correlation between blood loss, blood transfusion and oncological outcome in prostate cancer patients treated with RP. Therefore, the association between higher blood loss or transfusion rate and cancer recurrence as described in other surgical treated tumor entities seems to be irrelevant in prostate cancer patients.

Physiological and genetic analysis of multiple sodium channel variants in a model of genetic absence epilepsy.

In excitatory neurons, SCN2A (NaV1.2) and SCN8A (NaV1.6) sodium channels are enriched at the axon initial segment. NaV1.6 is implicated in several mouse models of absence epilepsy, including a missense mutation identified in a chemical mutagenesis screen (Scn8a(V929F)). Here, we confirmed the prior suggestion that Scn8a(V929F) exhibits a striking genetic background-dependent difference in phenotypic severity, observing that spike-wave discharge (SWD) incidence and severity are significantly diminished when Scn8a(V929F) is fully placed onto the C57BL/6J strain compared with C3H. Examination of sequence differences in NaV subunits between these two inbred strains suggested NaV1.2(V752F) as a potential source of this modifier effect. Recognising that the spatial co-localisation of the NaV channels at the axon initial segment (AIS) provides a plausible mechanism for functional interaction, we tested this idea by undertaking biophysical characterisation of the variant NaV channels and by computer modelling. NaV1.2(V752F) functional analysis revealed an overall gain-of-function and for NaV1.6(V929F) revealed an overall loss-of-function. A biophysically realistic computer model was used to test the idea that interaction between these variant channels at the AIS contributes to the strain background effect. Surprisingly this modelling showed that neuronal excitability is dominated by the properties of NaV1.2(V752F) due to "functional silencing" of NaV1.6(V929F) suggesting that these variants do not directly interact. Consequent genetic mapping of the major strain modifier to Chr 7, and not Chr 2 where Scn2a maps, supported this biophysical prediction. While a NaV1.6(V929F) loss of function clearly underlies absence seizures in this mouse model, the strain background effect is apparently not due to an otherwise tempting Scn2a variant, highlighting the value of combining physiology and genetics to inform and direct each other when interrogating genetic complex traits such as absence epilepsy.

Comparison of prostate cancer volume measured by HistoScanning™ and final histopathological results.

PURPOSE: HistoScanning™ (HS) is an ultrasound-based tissue characterization technique with encouraging results in the detection of prostate cancer (PCa). The aim of this study was to evaluate the accuracy of total tumor volume measured by HS (TVHS) in patients with PCa. METHODS: In 148 patients with proven PCa, TVHS was measured prior to radical prostatectomy and compared with the total tumor volume in the final pathological report (TVP) using the rank-based spearman correlation test. Correlation was performed after stratification of the results by d'Amico risk categories, prostate volume, experience of HS examiner, distance of the ultrasound probe to the prostate (≤3.5 and >3.5 mm) and quality of initial HS. In addition, a re-analysis of HS data was performed by a single examiner and the TVHS from the unmodified HS data was acquired. RESULTS: TVP was approximately twofold higher compared to TVHS. Overall, there was no significant correlation (r s = -0.0083, p = 0.9) for the TVP and the TVHS. After adjusting for d'Amico risk categories, prostate volume, experience of examiner, distance of the ultrasound probe to the prostate and quality of initial HS, no significant correlation was found. After re-analyzing of all HS data by 1 examiner, the correlation remained not significant (r s = 0.039, p = 0.6). CONCLUSIONS: TVHS and TVP did not correlate in this cohort of patients. We cannot recommend the use of HS at least for imaging of the total tumor volume at this time. The controversial findings for prostate HS should initiate more studies to clarify these discrepancies.

Does HistoScanning™ predict positive results in prostate biopsy? A retrospective analysis of 1,188 sextants of the prostate.

PURPOSE: The role of HistoScanning™ (HS) in prostate biopsy is still indeterminate. Existing literature is sparse and controversial. To provide more evidence according to that important clinical topic, we analyzed institutional data from the Martini-Clinic, Prostate Cancer Center, Hamburg. METHODS: Patients who received prostate biopsy and who also received HS were included in the study cohort. A single examiner, blinded to pathological results, re-analyzed all HS data in accordance with sextants of the prostate. Each sextant was considered as an individual case. Corresponding results from biopsy and HS were analyzed. The area under the receiver-operating characteristic curve (AUC) for the prediction of a positive biopsy by HS was calculated. Furthermore, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were assessed according to different HS signal volume cutoffs (>0, >0.2 and >0.5 ml). RESULTS: Overall, 198 men were identified and 1,188 sextants were analyzed. The AUC to predict positive biopsy results by HS was 0.58. Sensitivity, specificity, PPV and NPV for HS to predict positive biopsy results per sextant, depending on different HS signal volume cutoffs (>0, >0.2 and >0.5 ml) were 84.1, 27.7, 29.5 and 82.9 %, 60.9, 50.6, 28.8 and 79.7 %, and 40.1, 73.3, 33.1 and 78.8 %, respectively. CONCLUSIONS: Positive HS signals do not accurately predict positive prostate biopsy results according to sextant analysis. We cannot recommend a variation of well-established random biopsy patterns or reduction of biopsy cores in accordance with HS signals at the moment.

[The interoperability of IIEF-5 with EPIC-26 : Sexual function after radical prostatectomy]. / Die Übertragbarkeit von IIEF-5 (International Index of Erectile Function 5 Item Short Form) auf EPIC-26 (Expanded Prostate Cancer Index Composite 26) : Sexuelle Funktion nach radikaler Prostatektomie.

BACKGROUND: In the past, the IIEF­5 (International Index of Erectile Function 5 ) was predominantly used to measure erectile function in prostate cancer patients. Following international developments, the domain "sexuality" of the EPIC-26 (Expanded Prostate Cancer Index Composite 26) is increasingly used in Germany. OBJECTIVE: The aim of this work is to create a practicable comparison of the domain "sexuality" of the EPIC-26 with the IIEF­5 for treatment in Germany. This is particularly necessary for the evaluation of historical patient collectives. MATERIALS AND METHODS: For the evaluation, 2123 patients with prostate cancer confirmed by biopsy from 2014-2017 who completed both the IIEF­5 and the EPIC-26 were considered. Linear regression analyses are calculated to convert IIEF­5 sum scores to EPIC-26 sexuality domain scores. RESULTS: The correlation between IIEF­5 and the EPIC-26 domain score "sexuality" was 0.74, suggesting a high degree of content convergence between the constructs measured. While the standard error of the predicted values is relatively small, the prediction intervals are very wide. For example, for the critical IIEF­5 value of 22, the predicted value is 78.88 with a 95% prediction interval of 55.09 to 102.66. CONCLUSION: IIEF­5 and the Sexuality scale of the EPIC-26 measure a similar construct. The analysis shows that conversion of individual values is associated with great uncertainty. However, at the group level, the observed EPIC-26 "sexuality" score could be predicted quite accurately. This opens up the possibility of comparing the erectile function of cohorts of patients/test persons, even if this was collected with different measuring instruments.

[Quality criteria in urology : How to obtain comparable results?] / Qualitätskriterien in der Urologie : Wie kann eine Vergleichbarkeit der Ergebnisse geschaffen werden?

The standardization of procedural flow and medical documentation increasingly allows further possibilities. The best-known example of process standardization is the centralized treatment of complex clinical pictures, while patient-reported outcome measurements (PROMs) enable standardized documentation. Using the example of prostate cancer, existing literature on the topic of quality optimization in medicine is discussed. The following key points are addressed: (1) Increasing use of standardized PROMs for outcome documentation. (2) The transfer of complex clinical pictures to dedicated specialized centers has been shown to increase the quality of patient care as long as standardized PROMs are used. (3) Healthcare policymakers benefit from the use of PROMs and increasingly pursue a "value-based healthcare" approach.

Investigation of reaction force magnitude and orientation during supine thoracic thrust manipulation applied to intervertebral and costovertebral regions.

BACKGROUND: Spinal manipulative techniques are commonly used in manual therapies but quantified descriptive and reliability data are lacking considering supine thoracic thrust manipulation. OBJECTIVES: The purpose of this study is to explore and compare kinetic parameters during supine thoracic thrust manipulation performed at two different thoracic regions. Intra-rater task repeatability and influence of practitioners were estimated. DESIGN: Exploratory and agreement study. METHODS: Kinetic parameters were assessed by examining reaction force magnitude and orientation (on the basis of the zenithal angle) using force platforms. Manipulative procedure (consisting in the application of 3 preloads followed by one thrust adjustment) at both intervertebral and costovertebral region was performed by different practitioners at three sessions. Application of thrust was allowed for experienced practitioners only. Preload force, peak force magnitude and vector force orientation were compared between anatomical sites, sessions and practitioners, and bias with limit of agreement were estimated. RESULTS: Repeatability analysis showed that practitioners achieved similar preload and peak force independent of the session, with comparable force orientation. Differences between practitioners were observed for preload and peak force but not regarding the zenithal angle during the thrust phase. CONCLUSIONS: The present study is the first that explores kinetic parameters for supine thoracic thrust manipulation applied on two different regions of the thorax. Results confirm consistency of performance among practitioners for supine manipulative techniques at intervertebral and costovertebral region. While task repeatability was confirmed, several differences were observed between practitioners. Further investigations would examine velocity, acceleration and potential neurophysiological effect of such manipulative technique.

Outcome measures in perinatal medicine - pH or BE. The thresholds of these parameters in term infants.

BACKGROUND: Hypoxia and severe foetal acidosis may lead to cerebral injuries and multi-organ failure. Base excess (BE) and actual pH determined in umbilical artery (UA) blood are valid parameters to measure (metabolic) acidosis. Until now there is no consensus worldwide as to which of the two parameters should preferably be used and which thresholds should be applied: the thresholds 7.000, 7.100 and 7.200 are discussed for pH,UA and - 16.0 mmol/l for BE,UA, respectively. The aim of this study was to redefine these thresholds for term infants taking into account the entire spectrum of complications in early neonatal life and to compare the diagnostic power of both variables under investigation. METHODS: 512 foetuses all with a pH,UA < 7.100 were enrolled in this retrospective study. In this paper only term infants (n = 398) without major malformations were analysed. In order to quantify foetal morbidity and mortality an asphyxia complication (AC) score was designed using the Apgar score after 1 min and all possible complications encountered in the neonate until discharge (mainly from the NICU). Routine acid-base (AB) measurements (Radiometer, Copenhagen) were available (UA and UV blood) in nearly all cases. In this context, BE was corrected to 100 % oxygen-saturation (BE (oxy.)) using the (calculated) actual saturation (BE (act.oxy.)) of haemoglobin in each case because sO2 (%) becomes very low in severe acidosis. Oxygen saturation (%) was determined according to Ruiz et al. using haemoglobin F. Matched pairs (pH,UA > 7.10) were defined using the variables (i) gestational age, (ii) birth weight (+/- 100 g), (iii) sex, and (iv) parity. RESULTS: Analysing term infants with a definitely good outcome (n = 389) led to the following AB variables in UA blood: 10 % had a pH < 7.000; the lowest pH was 6.717, the highest pCO2 was 118 mmHg and the lowest BE(oxy.) amounted to - 32.4 mmol / l. 90 % of these neonates had an oxygen saturation that was still > 3.0 %. However, early neonatal morbidity due to hypoxia and acidosis was remarkable; therefore these AB variables could not serve as thresholds. Relying on clinical criteria, no acidotic morbidity was accepted, except for respiration disorders in early neonatal live (C score) together with Apgar scores down to 4 after one minute. This led to an AC score of < or = 8 (n = 378). In this group the lowest pH,UA was 6.890 and BE(oxy.) was - 25.1 mmol / l, respectively. In order to have a "buffer zone" of 10 % in each variable distribution (10 (th) / 90 (th) percentile). the following thresholds could be evaluated in UA blood: pH: 7.000, pCO2: 84 mmHg, sO2: 3.0 % and BE(oxy.) - 20.0 mmol/l. Only 13 (matched) neonates suffered from an AC score > 8 and had all pH values > or = 7.100 (3.3 % overlap). These foetuses seemed to be not at risk concerning injuries and severe complications. In UA blood the actual pH always offers closer correlations with the AC score when compared with the BE (oxy.) value. CONCLUSION: Thresholds in UA blood for pH, pCO2, sO2 and BE(oxy.) in term-infants are: 7.000, 84 mmHg, 3.0 % and - 20 mmol/l, respectively. Delivery of an otherwise healthy baby without getting in touch with these thresholds seems to be safe both for the baby and the obstetrician. In addition, severe neonatal depression (Apgar 1 min: 0 and 1) is usually avoided (0 / 398). BE(oxy.) does not offer a higher diagnostic power when compared with actual pH.

Prediction of the drilling path to surgically pin the femoral neck from the spatial location of pelvic and femoral anatomical landmarks: A cadaver validation study.

Several clinical applications rely on accurate guiding information when drilling along the femoral neck (e.g., pin insertion in case of neck fracture). Currently, applications rely on real-time X-ray imaging, which results in irradiation issues for the surgeon conducting the operation. The goal of this paper was to develop an X-ray-free method that would allow for a pathway to be drilled between the lateral aspect of the femoral diaphysis (the so-called piercing point), the femoral neck and the head centres. The method is based on on-the-fly computational predictions relying on a biomechanical database that includes morphological data related to the femoral neck and head and various palpable anatomical landmarks located on the pelvis and the femoral bone. From the spatial location of the anatomical landmarks, scalable multiple regressions allow for the prediction of the most optimal drilling pathway. The method has been entirely validated using in vitro experiments that reproduce surgical conditions. Further, a surgical ancillary prototype that integrates the method of guiding the pin drilling has been developed and used during in vitro and in situ validation using nine hip joints. Pin insertion was controlled after drilling using medical imaging and show successful result for each of the nine trials. The mean accuracy of the estimated hip joint centre and neck orientation was 6.0 ± 2.8mm and 7.1 ± 3.8°, respectively.

Prognostic value of the new Grade Groups in Prostate Cancer: a multi-institutional European validation study.

BACKGROUND: We aimed to assess the prognostic relevance of the new Grade Groups in Prostate Cancer (PCa) within a large cohort of European men treated with radical prostatectomy (RP). METHODS: Data from 27 122 patients treated with RP at seven European centers were analyzed. We investigated the prognostic performance of the new Grade Groups (based on Gleason score 3+3, 3+4, 4+3, 8 and 9-10) on biopsy and RP specimen, adjusted for established clinical and pathological characteristics. Multivariable Cox proportional hazards regression models assessed the association of new Grade Groups with biochemical recurrence (BCR). Prognostic accuracies of the models were assessed using Harrell's C-index. RESULTS: Median follow-up was 29 months (interquartile range, 13-54). The 4-year estimated BCR-free survival (bRFS) for biopsy Grade Groups 1-5 were 91.3, 81.6, 69.8, 60.3 and 44.4%, respectively. The 4-year estimated bRFS for RP Grade Groups 1-5 were 96.1%, 86.7%, 67.0%, 63.1% and 41.0%, respectively. Compared with Grade Group 1, all other Grade Groups based both on biopsy and RP specimen were independently associated with a lower bRFS (all P<0.01). Adjusted pairwise comparisons revealed statistically differences between all Grade Groups, except for group 3 and 4 on RP specimen (P=0.10). The discriminations of the multivariable base prognostic models based on the current three-tier and the new five-tier systems were not clinically different (0.3 and 0.9% increase in discrimination for clinical and pathological model). CONCLUSIONS: We validated the independent prognostic value of the new Grade Groups on biopsy and RP specimen from European PCa men. However, it does not improve the accuracies of prognostic models by a clinically significant margin. Nevertheless, this new classification may help physicians and patients estimate disease aggressiveness with a user-friendly, clinically relevant and reproducible method.

[Primary Tumour Treatment in Patients with Metastatic Prostate Cancer]. / Therapie des Primärtumors beim metastasierten Prostatakarzinom.

Prostate cancer (PCa) with distant metastasis at diagnosis (M1) is associated with an unfavourable prognosis. Therefore, according to current treatment guidelines, local treatment (radical prostatectomy or irradiation of the prostate gland) is not recommended in men with M1 disease. However, M1 patients inevitably develop castration-resistant disease progression leading to local complications in half of men. Hence, local treatment, if reconcilable with a good quality of life, would have the potential to prevent future local symptoms. Furthermore, evolving data from genomic studies suggest that local treatment might have the potential to reduce further spread of lethal cancer clones by eliminating the primary tumour or nodal metastasis. This leads to the experimental concept of cytoreductive preventative local treatment. According to large US-American and German cancer registries, there is a growing body of evidence pointing towards a survival benefit for M1 patients who receive local treatment in combination with systemic therapy. These data provide the basis to generate the hypothesis of a better response to systemic tumour therapy in metastatic prostate cancer, which would be the rationale for prospective randomised trials, but would not suffice for a treatment recommendation to be given. While there is a dynamic paradigm shift in the systemic treatment of hormone-naive M1 prostate cancer, local treatment for primary tumours will only have a chance to be established in this indication if prospective randomised trials are successfully completed.

Birth Control in Clinical Trials: Industry Survey of Current Use Practices, Governance, and Monitoring.

The Health and Environmental Sciences Institute (HESI) Developmental and Reproductive Toxicology Technical Committee sponsored a pharmaceutical industry survey on current industry practices for contraception use during clinical trials. The objectives of the survey were to improve our understanding of the current industry practices for contraception requirements in clinical trials, the governance processes set up to promote consistency and/or compliance with contraception requirements, and the effectiveness of current contraception practices in preventing pregnancies during clinical trials. Opportunities for improvements in current practices were also considered. The survey results from 12 pharmaceutical companies identified significant variability among companies with regard to contraception practices and governance during clinical trials. This variability was due primarily to differences in definitions, areas of scientific uncertainty or misunderstanding, and differences in company approaches to enrollment in clinical trials. The survey also revealed that few companies collected data in a manner that would allow a retrospective understanding of the reasons for failure of birth control during clinical trials. In this article, suggestions are made for topics where regulatory guidance or scientific publications could facilitate best practice. These include provisions for a pragmatic definition of women of childbearing potential, guidance on how animal data can influence the requirements for male and female birth control, evidence-based guidance on birth control and pregnancy testing regimes suitable for low- and high-risk situations, plus practical methods to ascertain the risk of drug-drug interactions with hormonal contraceptives.

[Systematic analysis of treatment results as a quality control instrument using the example of a large European center]. / Systematische Ergebnisanalyse als Qualitätsinstrument am Beispiel des größten europäischen Zentrums.

BACKGROUND: Measurement of the quality of treatment has become increasingly important in hospitals. An easy and practical solution in data acquisition is the Patient-Reported Outcome Measurement (PROM). METHODS: In this article the historical development, general conditions, and difficulties of using the outcome measurement in our patients are describe. In addition, we illustrate the wide range of benefits due to our survey. Based on data from 2008-2013, the functional and oncological results of treatment in our clinic are shown. The main focus lies on the PROMs, e.g., urinary continence evaluated with the daily pad use, sexual function with the IIEF-5 questionnaire, and postoperative complications. CONCLUSIONS: A systematic and standardized outcome measurement may help to improve the quality of treatment, provides factual information for patients, and supports medical development.

["Expanded prostate cancer index composite" (EPIC-26): Results of functional treatment in patients with localized prostate cancer]. / "Expanded prostate cancer index composite" (EPIC-26) : Funktionelles Behandlungsergebnis bei Patienten mit lokalisiertem Prostatakarzinom.

The standardized collation of the quality of treatment is a subject of discussion both nationally and internationally. This article presents the work of the International Consortium for Health Outcomes Measurement (ICHOM) and the validated German translation of the expanded prostate cancer index composite (EPIC-26). This questionnaire allows a standardized interdisciplinary collation of the quality of treatment for all therapy modalities of localized prostate cancer. Use of the ICHOM standard set and the EPIC-26 achieves a possibility for comparison of each form of therapy with respect to the curative success and the effect on health and quality of life of patients.

[DaVinci robot-assisted laparoscopic prostatectomy: benefit for obese men? - A matched-pair analysis]. / Roboterassistierte radikale Prostatektomie : Vorteil bei adipösen Männern? - Eine Matched-pair-Analyse.

BACKGROUND: Open radical retropubic prostatectomy (RRP) in obese patients (BMI ≥30) is associated with increased perioperative morbidity. The aim of the study was to evaluate the possible benefit of DaVinci robotic-assisted laparoscopic prostatectomy (RARP) compared to RRP in obese patients. PATIENTS AND METHODS: We identified 255 patients with a localized prostate cancer (PCa) and BMI ≥30 treated with radical prostatectomy from January 2009 to December 2011. To adjust for risk factors of increased perioperative morbidity (nerve-sparing, pelvic lymph node dissection, prostate volume), a propensity score-based matching was performed between RRP and RARP (n=115 each group). Both groups were compared by taking into consideration histopathological outcomes as well as peri- and postoperative (30 days) morbidity. RESULTS: There were no differences in histopathological characteristics (pT/pN-stage, Gleason score, R-stage; all p>0.05) in both groups. Mean blood loss (276 ml vs. 937 ml), transfusion rate (0.9% vs. 8.7%) and 30-day complications according to the Clavien classification system (Clavien ≥ 2; 9.5% vs. 22.6%) were decreased in RARP (all p<0.05). In a multivariate logistic regression model, RARP vs. RRP was associated with a significantly reduced risk of a Clavien ≥ 2 complication during follow-up (OR 0.3; p= 0.0047). Recovery of continence was significantly better for RARP patients after 3 months (p= 0.02). There was no difference in erectile function 12 months postoperatively. CONCLUSION: Our findings of decreased transfusion and complication rates and a trend of better early recovery of continence in RARP should be considered in obese patients (BMI >30) scheduled for radical prostatectomy.

Adherence of the indication to European Association of Urology guideline recommended pelvic lymph node dissection at a high-volume center: Differences between open and robot-assisted radical prostatectomy.

PURPOSE: Contemporary adherence of the indication to European Association of Urology (EAU) guideline recommendation for pelvic lymph node dissection (PLND) at either open (ORP) or robot-assisted radical prostatectomy (RARP) at a high-volume center is unknown. To assess guideline recommended and observed PLND rates in a high-volume center cohort. METHODS: We relied on the Martini-Clinic database and focused on patients treated with either ORP or RARP, between 2010 and 2013. Actual performed PLND was compared to European Association of Urology (EAU) guideline recommendation defined by nomogram predicted risk of lymph node invasion >5%. Categorical and multivariable logistic regression analyses targeted two endpoints: 1) probability of guideline recommended PLND and 2) probability of no PLND, when not recommended by EAU guideline. RESULTS: Within 7868 PCa patients, adherence to EAU PLND guideline recommendation was 97.1% at ORP and 96.8% at RARP (p = 0.7). When PLND was not recommended, it was more frequently performed at RARP (71.6%) than at ORP (66.2%) (p = 0.002). Gleason score, PSA and number of positive biopsy cores were independent predictors for both either PLND when recommended, or no PLND when not recommended (all p < 0.05). Clinical tumor stage, age and surgical approach were also independent predictors for no PLND when not recommended (all p < 0.05). CONCLUSIONS: Adherence of the indication to EAU guideline recommended PLND is high at this high-volume center. Neither ORP nor RARP represent a barrier for PLND, when recommended. However, a high number of patients underwent PLND despite absence of guideline recommendation. Possible staging advantages and PLND related complications needs to be individually considered, especially, when LNI risk is low.

Prime region subsite specificity characterization of human cathepsin D: the dominant role of position 128.

In order to contribute to our understanding of cathepsin D (CatD) active site specificity, two series of chromogenic octapeptides with systematic substitutions in positions P2' and P3' were synthesized. This panel was characterized with native human liver cathepsin D (nHuCatD) and yielded information concerning specificity trends within the S2' and S3' subsites. The pepstatin inhibited crystal structure of nHuCatD (Baldwin et al., 1993) was then utilized in conjunction with these subsite preference data to identify residues suspected of contributing to "prime" side subsite specificity. These residues were targeted for site-directed mutagenesis using the re-engineered recombinant model, "short" pseudocathepsin D (Beyer & Dunn, 1996). As a result of these analyses it was determined that prime region subsites do contribute to the unique specificity of human CatD. Furthermore, it was ascertained that the poly-proline loop does not have an active role in S3' subsite specificity. Lastly, it appears that Ile128 has a dominant role on S2' subsite specificity whereas Val130 does not.

A novel recombinant single-chain hepatitis C virus NS3-NS4A protein with improved helicase activity.

Hepatitis C virus (HCV) nonstructural protein 3 (NS3) has been shown to possess protease and helicase activities and has also been demonstrated to spontaneously associate with nonstructural protein NS4A (NS4A) to form a stable complex. Previous attempts to produce the NS3/NS4A complex in recombinant baculovirus resulted in a protein complex that aggregated and precipitated in the absence of nonionic detergent and high salt. A single-chain form of the NS3/NS4A complex (His-NS4A21-32-GSGS-NS3-631) was constructed in which the NS4A core peptide is fused to the N-terminus of the NS3 protease domain as previously described (Taremi et al., 1998). This protein contains a histidine tagged NS4A peptide (a.a. 21-32) fused to the full-length NS3 (a.a. 3-631) through a flexible tetra amino acid linker. The recombinant protein was expressed to high levels in Escherichia coli, purified to homogeneity, and examined for NTPase, nucleic acid unwinding, and proteolytic activities. The single-chain recombinant NS3-NS4A protein possesses physiological properties equivalent to those of the NS3/NS4A complex except that this novel construct is stable, soluble and sixfold to sevenfold more active in unwinding duplex RNA. Comparison of the helicase activity of the single-chain recombinant NS3-NS4A with that of the full-length NS3 (without NS4A) and that of the helicase domain alone suggested that the presence of the protease domain and at least the NS4A core peptide are required for optimal unwinding activity.