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Background: During the last decade, arthroscopic procedures have been replacing open techniques in Haglund disease treatment because of their considerable advantages. Endoscopic calcaneoplasty is a technique that allows resection of posterosuperior calcaneal exostosis and retrocalcaneal bursitis. The objective of this article was to describe this technique and report its clinical and subjective outcome., Methods: A retrospective study was performed of consecutive patients undergoing endoscopic Haglund resection surgery between July 2014 and March 2020 at a single academic institution. All patients were surveyed in person about the level of pain (visual analog scale), its location (central, lateral, medial or diffuse), its relation with rest, or physical activity. Clinical evaluation was assessed using the hindfoot scale designed by the American Orthopaedic Foot & Ankle Society (AOFAS)., Results: In this study, 14 endoscopic calcaneoplasties were performed in 14 patients, with an average follow-up of 40 months. The visual analog scale score improved from a preoperative average value of 9.07 to 1.8 after surgery ( P  > .0001). The AOFAS scale rose from 38.7 before surgery to 94.6 postoperative ( P  > .0001). Good subjective results were observed in 12 patients (85.7%), and all of them would have surgery again. There were no wound complications or infections. No patient required reoperation., Conclusion: In this relatively small cohort, we found that endoscopic calcaneoplasty was associated with good clinical and subjective results with few complications., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article. Disclosure forms for all authors are available online.

Aim: Functional Management and Recovery is a standardized Psychoeducational Intervention, derived from "Integro", an effective salutogenic-psychoeducational intervention for people in recovery journey, designed to improve recovery and functioning of individuals with psychotic disorders in Psychiatric Residential Facilities (PRFs). The aim of this study is to evaluate the primary and secondary outcomes of this intervention elaborated specifically for PRFs where evidence based structured interventions seem rare and desirable., Methods: 66 individuals with psychotic disorders were recruited in 9 PRFs dislocated in the North, Center and South Italy and 63 underwent a multicenter follow-up study with a two time-point evaluation (t0, pre-treatment and t1, 6 months; ). At each time point, social functioning was assessed as primary outcome by the Personal and Social Performance scale (PSP); furthermore, psychopathological status was assessed by Brief Psychiatric Rating Scale (BPRS), Recovery by Recovery Assessment Scale (RAS), Cognitive Functioning by Repeatable Battery for the Assessment of Neuropsychological Status (RBANS), Stress management by Stress-Scale, Cognitive Flexibility by Modified Five-Point Test (M-FPT), Emotional Intelligence by Emotional Intelligence Index (EI-I), the PRF Atmosphere and the Opinion of users about the PFR by an ad hoc questionnaire. The Abilities Knowledge, the Utility and Pleasantness of sessions were measured by an ad hoc list of items., Results: 63 individuals out of 66, 52 (82,5%) affected by schizophrenia and 11 (17,5%) by bipolar I disorder with psychotic symptoms according to DSM-5-TR completed the study. At the end of the study, 43 (68,3%) were male, 57 (90.5%) were single, 5 (7.9%) engaged, 1 (1.6%) married; 45 (71.4%) unemployed. The total scores of PSP, RAS, BPRS, BANS, Stress management, Abilities Knowledge, Utility and Pleasantness of sessions showed a statistically significant improvement at t1 vs. t0. Two sub-scales out of 5 of M-FPT showed a statistically significant improvement. The Emotional Intelligence, the Unit Atmosphere and the Opinion of Users about PFR improved without statistical significance. Six months after the end of the follow-up study 22 individuals of the sample were dismissed with a very high turnover., Conclusions: After a six-month follow-up (a short period of time), these results showed improvement in functioning, the primary outcome, as well as in the following secondary outcome variables: RAS, BPRS, BANS, Stress management, Abilities Knowledge, two sub-scales out of 5 of M-FPT, Utility and Pleasantness of sessions. Overall, a remarkable impact of psychoeducational structured intervention on the key Recovery variables is observed. Further studies are needed to address extent and duration of these improvements., (© 2024. The Author(s).)

Introduction: The modified 5-item frailty index (mFI-5) has been recently proposed as a useful tool for predicting postoperative complications in orthopedic surgery. We aimed to analyze the utility of this score in predicting complications and reoperations after hallux valgus (HV) deformity surgery., Methods: 551 patients undergoing percutaneous HV corrective surgery were retrospectively reviewed. The mFI-5 was calculated based and patients were categorized in three groups: 1) non-frail: patients without any of the 5 comorbidities, 2) pre-frail: patients with one comorbidity and 3) frail: patients with two or more comorbidities. Complications and surgical reoperations were recorded., Results: In the study period 772 percutaneous surgeries were performed to correct HV deformity, 551 patients were included with a median age of 60 (IQR 48-70). Three hundred eighty-nine patients were non-frail (70.6%), 132 were pre-frail (23.9%) and 30 were frail (5.4%). 75 patients suffered complications (13.6%). Even though the rate of complications was higher in frailty patients (23.3%) compared with pre-frail (13.6%) and non frail (12.8%), no significant differences were observed among groups. 48 patients required reoperation (8.7%) but the rate of reoperations among frailty groups was not significantly different (P=.11). Frailty patients had worse AOFAS scores at final follow up (P=.011)., Conclusion: The mFI-5 was not useful to predict postoperative complications and reoperations after hallux valgus corrective surgery. Therefore, other factors should be considered when analyzing the risk of complications after HV corrective surgery., (Copyright © 2023 SECOT. Publicado por Elsevier España, S.L.U. All rights reserved.)

Introduction: The modified 5-item frailty index (mFI-5) has been recently proposed as a useful tool for predicting postoperative complications in orthopedic surgery. We aimed to analyze the utility of this score in predicting complications and reoperations after hallux valgus (HV) deformity surgery., Methods: 551 patients undergoing percutaneous HV corrective surgery were retrospectively reviewed. The mFI-5 was calculated based and patients were categorized in three groups: (1) non-frail: patients without any of the 5 comorbidities, (2) pre-frail: patients with one comorbidity and (3) frail: patients with two or more comorbidities. Complications and surgical reoperations were recorded., Results: In the study period 772 percutaneous surgeries were performed to correct hallux valgus deformity, 551 patients were included with a median age of 60 (IQR 48-70). 389 patients were non-frail (70.6%), 132 were pre-frail (23.9%) and 30 were frail (5.4%). 75 patients suffered complications (13.6%). Even though the rate of complications was higher in frailty patients (23.3%) compared with pre-frail (13.6%) and non frail (12.8%), no significant differences were observed among groups. 48 patients required reoperation (8.7%) but the rate of reoperations among frailty groups was not significantly different (p=0.11). Frailty patients had worse AOFAS scores at final follow up (p=0.011)., Conclusion: The mFI-5 was not useful to predict postoperative complications and reoperations after hallux valgus corrective surgery. Therefore, other factors should be considered when analyzing the risk of complications after HV corrective surgery., (Copyright © 2023 SECOT. Publicado por Elsevier España, S.L.U. All rights reserved.)

CT urography is a single term used to refer to different scanning protocols that can be applied for a number of clinical indications. If, on the one hand, this highlights the role of the radiologist in deciding the most suitable technique to perform according to the patient's needs, on the other hand, a certain confusion may arise due to the different technical and clinical variables that have to be taken into account. This has been well demonstrated by a previous work based on an online questionnaire administered to a population of Italian radiologists that brought out similarities as well as differences across the national country. Defining precise guidelines for each clinical scenario, although desirable, is a difficult task to accomplish, if not even unfeasible. According to the prementioned survey, five relevant topics concerning CT urography have been identified: definition and clinical indications, opacification of the excretory system, techniques, post-processing reconstructions, and radiation dose and utility of dual-energy CT. The aim of this work is to deepen and share knowledge about these main points in order to assist the radiology in the daily practice. Moreover, a synopsis of recommendations agreed by the Italian board of genitourinary imaging is provided., (© 2023. Italian Society of Medical Radiology.)

Waxy (WX) and high-amylose (HA) wheat flours have interesting functional and/or nutritional characteristics, but low technological properties compared to regular wheat. Here a set of three wheat lines, having different amylose content but sharing the same varietal background, were compared to shed light on the role of the amylose/amylopectin ratio on the protein conformational changes that lead to gluten formation. Despite the absence of differences in their protein profile, as also confirmed by thiolomic approaches, both WX and HA lines developed a weaker gluten than the control sample. The altered amylose/amylopectin ratio exerts a matrix effect establishing a competition for water with proteins, leading to a different protein structure and three-dimensional organization of the gluten network. These results add a piece to the understanding of the molecular aspects that oversee matrix effects on gluten formation in wheat, which description can be helpful for a rational optimization of the transformation process., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Aim: To assess urologists’ proficiency in the interpretation of multiparametric magnetic resonance imaging (mpMRI). Materials and Methods: Twelve mpMRIs were shown to 73 urologists from seven Italian institutions. Responders were asked to identify the site of the suspicious nodule (SN) but not to assign a PIRADS score. We set an a priori cut-off of 75% correct identification of SN as a threshold for proficiency in mpMRI reading. Data were analyzed according to urologists’ hierarchy (UH; resident vs. consultant) and previous experience in fusion prostate biopsies (E-fPB, defined as <125 vs. ≥125). Additionally, we tested for differences between non-proficient vs. proficient mpMRI readers. Multivariable logistic regression analyses (MVLRA) tested potential predictors of proficiency in mpMRI reading. Results: The median (IQR) number of correct identifications was 8 (6−8). Anterior nodules (number 3, 4 and 6) represented the most likely prone to misinterpretation. Overall, 34 (47%) participants achieved the 75% cut-off. When comparing consultants vs. residents, we found no differences in terms of E-fPB (p = 0.9) or in correct identification rates (p = 0.6). We recorded higher identification rates in urologists with E-fBP vs. their no E-fBP counterparts (75% vs. 67%, p = 0.004). At MVLRA, only E- fPB reached the status of independent predictor of proficiency in mpMRI reading (OR: 3.4, 95% CI 1.2−9.9, p = 0.02) after adjusting for UH and type of institution. Conclusions: Despite urologists becoming more familiar with interpretation of mpMRI, their results are still far from proficient. E-fPB enhances the proficiency in mpMRI interpretation.

Background: Gas in the renal excretory system is described as a consequence of recent procedures but it can be a sign of severe conditions such as fistulas or infections; however, gas-containing renal stones are only rarely encountered., Purpose: To describe the association of gas-containing renal stones and urinary tract infection., Material and Methods: We performed a retrospective evaluation of the clinical and imaging findings in a series of six patients with gas-containing renal stones and compared our findings with those of patients with gas-containing renal stones reported in the literature. Urine and stone cultures were used as a diagnostic standard for urinary tract infection., Results: Including the present series, there is a total of 21 patients with gas-containing renal stones in the literature. Based on clinical presentation, urinary tract infection could be suspected in 10 (57%) patients, while urine and/or stone cultures showed infection in 18 of 19 (95%) patients, with only one case with no bacterial growth in both (5%); in the remaining patient the information was not available., Conclusion: Gas-containing renal stones are a rarely reported entity usually diagnosed with computed tomography. They are a radiological sign often associated with urinary tract infection that can also be encountered in patients with non-specific renal symptoms.

Purpose: To determine the clinical, pathological, and radiological features, including the Vesical Imaging-Reporting and Data System (VI-RADS) score, independently correlating with muscle-invasive bladder cancer (BCa), in a multicentric national setting., Method and Materials: Patients with BCa suspicion were offered magnetic resonance imaging (MRI) before trans-urethral resection of bladder tumor (TURBT). According to VI-RADS, a cutoff of ≥ 3 or ≥ 4 was assumed to define muscle-invasive bladder cancer (MIBC). Trans-urethral resection of the tumor (TURBT) and/or cystectomy reports were compared with preoperative VI-RADS scores to assess accuracy of MRI for discriminating between non-muscle-invasive versus MIBC. Performance was assessed by ROC curve analysis. Two univariable and multivariable logistic regression models were implemented including clinical, pathological, radiological data, and VI-RADS categories to determine the variables with an independent effect on MIBC., Results: A final cohort of 139 patients was enrolled (median age 70 [IQR: 64, 76.5]). MRI showed sensitivity, specificity, PPV, NPV, and accuracy for MIBC diagnosis ranging from 83-93%, 80-92%, 67-81%, 93-96%, and 84-89% for the more experienced readers. The area under the curve (AUC) was 0.95 (0.91-0.99). In the multivariable logistic regression model, the VI-RADS score, using both a cutoff of 3 and 4 (P < .0001), hematuria (P = .007), tumor size (P = .013), and concomitant hydronephrosis (P = .027) were the variables correlating with a bladder cancer staged as ≥ T2. The inter-reader agreement was substantial (k = 0.814)., Conclusions: VI-RADS assessment scoring proved to be an independent predictor of muscle-invasiveness, which might implicate a shift toward a more aggressive selection approach of patients' at high risk of MIBC, according to a novel proposed predictive pathway., (© 2022. The Author(s).)

The recent socio-economic situation requires producers to change the composition of basic foods. The aim of this study was to assess the technological properties of wheat flour enriched with cricket powder (CP) (at 5%, 10%, and 20% levels) for the development of bread and pasta. The hydration (i.e., water absorption capacity, oil absorption capacity, water absorption index, water solubility index, and swelling power), foaming (i.e., foaming capacity and stability),emulsifying (emulsifying activity and emulsion stability), and rheological (during gluten aggregation, mixing, extension, and leavening) properties were investigated. Finally, bread and fresh pasta were prepared and characterized. Emulsifying activity, stability, and foaming capacity decreased in the presence of CP, whereas foaming stability and water solubility increased. The results on dough rheology highlighted the need to increase the amount of water, and to decrease the mixing and leavening time, to keep an acceptable bread volume. Indeed, 10% CP enrichment led to a product characterized by a similar volume and crumb hardness to the control (wheat flour). Despite the decrease in extensibility caused by CP, it was possible to produce fresh pasta enriched with CP, with the best cooking behavior obtained at a 5% replacement level.

Computed tomography-urography is currently the imaging modality of choice for the assessment of the whole urinary tract, giving the possibility to detect and characterize benign and malignant conditions. In particular, computed tomography-urography takes advantage from an improved visualization of the urinary collecting system due to acquisition of delayed scan obtained after excretion of intravenous contrast medium from the kidneys. Nevertheless, the remaining scans are of great help for identification, characterization, and staging of urological tumors. Considering the high number of diseases, urinary segment potentially involved and patients' features, scanning protocols of computed tomography-urography largely vary from one clinical case to another as well as selection and previous preparation of the patient. According to the supramentioned considerations, radiation exposure is also of particular concern. Italian radiologists were asked to express their opinions about computed tomography-urography performance and about its role in their daily practice through an online survey. This paper collects and summarizes the results., (© 2022. Italian Society of Medical Radiology.)

Despite being considered a climate-resilient crop, sorghum is still underutilized in food processing because of the limited starch and protein functionality. For this reason, the objective of this study was to investigate the effect of sprouting time on sorghum functional properties and the possibility to exploit sprouted sorghum in bread making. In this context, red sorghum was sprouted for 24, 36, 48, 72, and 96 h at 27 °C. Sprouting time did not strongly affect the sorghum composition in terms of total starch, fiber, and protein contents. On the other hand, the developed proteolytic activity had a positive effect on oil-absorption capacity, pasting, and gelation properties. Conversely, the increased α-amylase activity in sprouted samples (≥36 h) altered starch functionality. As regards sorghum-enriched bread, the blends containing 48 h-sprouted sorghum showed high specific volume and low crumb firmness. In addition, enrichment in sprouted sorghum increased both the in vitro protein digestibility and the slowly digestible starch fraction of bread. Overall, this study showed that 48 h-sprouted sorghum enhanced the bread-making performance of wheat-based products.

The aim of the study is to evaluate the performance of a simplified ABLATE score (sABLATE) in predicting complications and outcome with respect to RENAL, mRENAL, and ABLATE scores. This study included 136 renal lesions in 113 patients (M:F ratio = 2.5; mean age 70.8 years). 98 tumors underwent cryoablation at San Raffaele hospital between 01/2015 and 03/2020, while 37 underwent microwave ablation at San Paolo or Policlinico hospitals between 07/2016 and 03/2020. RENAL, mRENAL, ABLATE, and sABLATE scores were calculated using pre-procedural imaging. Data regarding complications and follow-up were registered. Mann-Whitney U test, ROC analyses, and logistic regression analyses were used for complications. Cox-regression analyses were performed for outcome. Mean tumor diameter was 23.2 mm. Mean and median RENAL, mRENAL, ABLATE, and sABLATE scores were 6.8 and 7, 6.9 and 7, 5.3, and 5, and 3.5 and 3, respectively. During a mean follow-up of 21.9 months (range 1-73), we registered 7 complications, 3 cases of residual disease, and 10 local tumor progressions. Mann-Whitney U test p values for complications for RENAL, mRENAL, ABLATE, and sABLATE were 0.51, 0.49, 0.66, and 0.056, respectively. ROC analyses for complications showed an AUC for RENAL, mRENAL, ABLATE, and sABLATE of 0.57, 0.57, 0.55, and 0.71, respectively. Regarding outcome, HR and p values of Cox-regression analyses were 1.30 and 0.36 for RENAL, 1.33 and 0.35 for mRENAL, 2.16 and 0.01 for ABLATE, 2.29 and 0.004 for sABLATE. sABLATE was the only score close to significance for complications, representing a progress even if not definitive. Regarding outcome, ABLATE confirmed its value, and sABLATE maintained validity despite being a simplification., (© 2021. Springer Science+Business Media, LLC, part of Springer Nature.)

Despite the interest in stone-milling, there is no information on the potential advantages of using the resultant wholegrain flour (WF) in bread-making. Consequently, nutritional and technological properties of WFs obtained by both stone- (SWF) and roller-milling (RWF) were assessed on four wheat samples, differing in grain hardness and pigment richness. Regardless of the type of wheat, stone-milling led to WFs with a high number of particles ranging in size from 315 to 710 μm), whereas RWFs showed a bimodal distribution with large (>1000 μm) and fine (<250 μm) particles. On average, the milling system did not affect the proximate composition and the bioactive features of WFs. The gluten aggregation kinetics resulted in similar trends for all SWFs, with indices higher than for RWFs. The effect of milling on dough properties (i.e., mixing and leavening) was sample dependent. Overall, SWFs produced more gas, resulting in bread with higher specific volume. Bread crumb from SWF had higher lutein content in the wheat cv rich in xanthophylls, while bread from RWF of the blue-grained cv had a moderate but significantly higher content in esterified phenolic acids and total anthocyanins. In conclusion, there was no relevant advantage in using stone- as opposed to roller-milling (and vice versa).

Background: Pre-harvest sprouting of wheat is viewed negatively because of the high level of enzymatic activity, which leads to a deterioration in the bread-making performance of the related flours. On the other hand, improvements in bread properties (i.e. volume and crumb softness) are reported when sprouted wheat under controlled conditions is used in mixtures with a conventional unsprouted flour. However, knowledge about the effects of sprouting on gluten functionality and its relationship with bread features is still limited, especially in the case of whole wheat flour., Results: Under the conditions applied in this study (48 h, 20 °C and 90% relative humidity), proteins of sprouted wheat were still able to aggregate, even if changes in gluten aggregation kinetics suggested gluten weakening. On the other hand, sprouting led to an increase in gluten stretching ability, suggesting an increase in dough extensibility. In the dough system, sprouting was responsible for a decrease in water absorption, development time, and stability during mixing. However, when the values for development time and water absorption indicated by the Farinograph® were followed carefully, sprouting improved bread height (~20%), specific volume (~15%), and crumb softness (~200% after 24 h of storage), even when whole wheat flour was used., Conclusion: It is possible to produce bread with improved volume and crumb softness using whole wheat flour from sprouted kernels. Thus, sprouting can be exploited as a pre-treatment to improve the bread-making performance of fiber-enriched systems. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

RENAL score has been validated on predicting adverse events and relapses in percutaneous treatments of renal lesions. To better fit interventional issues a modified score (mRENAL) has been introduced, but the only difference from the RENAL score is on the dimensional parameter. However, it remains of surgical derivation while a specific interventional score is missing. This study aims to obtain a specific score (ABLATE) to better quantify the risk of complications and relapses in percutaneous kidney ablation procedures compared to the existing surgical scores. Taking inspiration from previous papers, a score was built to quantify the real difficulties faced in percutaneous treatment of renal lesions. The ABLATE score was used on 71 cryoablations to evaluate its predictivity of complications and relapses. Logistic regression was used to predict complication incidence; Cox-regression was used for relapses; ROC analysis was used to evaluate the accuracy of the different scores. Between January 2014 and November 2019, 71 lesions in 68 patients were treated. Overall, malignant histology was found in 62 lesions (87.3%). Mean and median RENAL, mRENAL, and ABLATE scores were 7.04 and 7, 7.19 and 7, and 5.11 and 4, respectively. Out of 71 treatments, we experienced 3 bleeding with anemia (4.2%), only 2 of which needed further treatment (2.82%). The mean and median RENAL, mRENAL, and ABLATE scores in those with complications were 7.66 and 7.01 (p = 0.69), 8.0 and 7.1 (p = 0.54), and 6.6 and 5.0 (p = 0.38), respectively. Out of 62 malignant lesions, we experienced 2 persistent and 6 recurrent lesions (3.2% and 8.4%, respectively). At Cox-regression analyses, mABLATE score outperformed both RENAL and mRENAL scores in predicting recurrences (HR 1.48; p < 0.001 vs. 1.41; p = 0.1 vs. 1.38: p = 0.07, respectively). The ABLATE score showed to be a better predictor of relapses than RENAL and mRENAL. The small number of complications conditioned a lack of statistic power on complications for all the scores. At the moment to quantify the risks in percutaneous kidney ablation procedures, surgical scores are used. A specific score better performs this task.

This research investigated the effect of sprouting on wheat bran. Bran from un-sprouted (BUW) and sprouted (BSW) wheat were characterized in terms of chemical composition, enzymatic activities, and hydration properties. In addition, the rheological properties (using GlutoPeak, Farinograph, Extensograph, and Rheofermentometer tests) and bread-making performance (color, texture, volume of bread) of wheat doughs enriched in bran at 20% replacement level were assessed. Sprouting process caused a significant decrease in phytic acid (~20%), insoluble dietary fiber (~11%), and water holding capacity (~8%), whereas simple sugars (~133%) and enzymatic activities significantly increased after processing. As regards the gluten aggregation kinetics, the BSW-blend profile was more similar to wheat than BUW-blend, indicating changes in the fiber and gluten interactions. BSW led to a worsening of the mixing and leavening properties, instead, no significant changes in extensibility were observed. Finally, BSW improved bread volume (~10%) and crumb softness (~52%). Exploiting bran from sprouted wheat might be useful to produce bread rich in fiber with enhanced characteristics.

Mitochondria participate in metabolism and signaling. They adapt to the requirements of various cell types. Publicly available expression data permit to study expression dynamics of genes with mitochondrial function (mito-genes) in various cell types, conditions and organisms. Yet, we lack an easy way of extracting these data for mito-genes. Here, we introduce the visual data mining platform mitoXplorer, which integrates expression and mutation data of mito-genes with a manually curated mitochondrial interactome containing ∼1200 genes grouped in 38 mitochondrial processes. User-friendly analysis and visualization tools allow to mine mitochondrial expression dynamics and mutations across various datasets from four model species including human. To test the predictive power of mitoXplorer, we quantify mito-gene expression dynamics in trisomy 21 cells, as mitochondrial defects are frequent in trisomy 21. We uncover remarkable differences in the regulation of the mitochondrial transcriptome and proteome in one of the trisomy 21 cell lines, caused by dysregulation of the mitochondrial ribosome and resulting in severe defects in oxidative phosphorylation. With the newly developed Fiji plugin mitoMorph, we identify mild changes in mitochondrial morphology in trisomy 21. Taken together, mitoXplorer (http://mitoxplorer.ibdm.univ-mrs.fr) is a user-friendly, web-based and freely accessible software, aiding experimental scientists to quantify mitochondrial expression dynamics., (© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Purpose: Radiofrequency and cryoablation (Cryo) are the most widely used techniques for the treatment of T1a renal tumors in non-surgical candidates, yet microwave ablation (MWA) has been gaining popularity. In this study, we tested the hypothesis that MWA has comparable safety and efficacy to Cryo in the treatment of selected T1a renal masses., Materials and Methods: A retrospective comparative analysis of two patient cohorts was carried out on 83 nodules in 72 consecutive patients treated using image-guided percutaneous ablation with either Cryo or MWA. Patient demographics, tumor histology and characteristics, technical success, procedure time, adverse events and complications, nephrometry score (mRENAL) and renal function were evaluated. Local recurrence was evaluated at 1, 6, 12 and 18-24 months., Results: Fifty-one nodules were treated with Cryo and 32 with MWA (44 and 28 patients, respectively). No statistical differences were observed following Cryo or MWA in median tumor size (p = 0.6), mRENAL (p = 0.1) or technical success (p = 0.8). Median procedure time was significantly lower using microwave ablation (p = 0.003). Median follow-up time was similar in the two groups (22 and 20 months, respectively). Occurrence of complications did not differ (Cryo 5/51, MWA 2/32; p = 0.57), and probability of complications or technical success adjusted for mRENAL did not reach statistical significance (p = 0.6). Renal function was preserved in all patients regardless of techniques. Disease recurrence was observed in 3/47 and in 1/30 treated nodules in the Cryo and MWA groups, respectively, without reaching statistical significance (p = 0.06)., Conclusion: In the patient population studied, MWA showed comparable safety and efficacy relative to Cryo., Level of Evidence: Level 3, Non-randomized cohort study.

The co-self-assembly of proteins and nucleic acids (NAs) produces complex biomolecular machines (e.g., ribosomes and telomerases) that represent some of the most daunting targets for biomolecular design. Despite significant advances in protein and DNA or RNA nanotechnology, the construction of artificial nucleoprotein complexes has largely been limited to cases that rely on the NA-mediated spatial organization of protein units, rather than a cooperative interplay between protein- and NA-mediated interactions that typify natural nucleoprotein assemblies. We report here a structurally well-defined synthetic nucleoprotein assembly that forms through the synergy of three types of intermolecular interactions: Watson-Crick base pairing, NA-protein interactions, and protein-metal coordination. The fine thermodynamic balance between these interactions enables the formation of a crystalline architecture under highly specific conditions., Competing Interests: The authors declare no competing financial interest.

Microtubules are central elements of the eukaryotic cytoskeleton that often function as part of branched networks. Current models for branching include nucleation of new microtubules from severed microtubule seeds or from γ-tubulin recruited to the side of a pre-existing microtubule. Here, we found that microtubules can be directly remodelled into branched structures by the microtubule-remodelling factor SSNA1 (also known as NA14 or DIP13). The branching activity of SSNA1 relies on its ability to self-assemble into fibrils in a head-to-tail fashion. SSNA1 fibrils guide protofilaments of a microtubule to split apart to form daughter microtubules. We further found that SSNA1 localizes at axon branching sites and has a key role in neuronal development. SSNA1 mutants that abolish microtubule branching in vitro also fail to promote axon development and branching when overexpressed in neurons. We have, therefore, discovered a mechanism for microtubule branching and implicated its role in neuronal development.

Background: The extensive use of multiparametric magnetic resonance imaging (mpMRI) has led to an even more widespread use of different targeted biopsy techniques and approaches. The best way of performing targeted biopsies and the effect of operator expertise have still to be defined., Objective: To compare the rate of detection of clinically significant prostate cancer (csPCa) of different mpMRI targeted approaches and to assess the role of operator expertise in the detection of csPCa., Design, Setting, and Participants: We included 244 consecutive patients who underwent both 12-core transrectal ultrasound (TRUS) biopsy and mpMRI targeted biopsy with either a cognitive biopsy (CB) or fusion biopsy (FB) approach during the same session between 2013 and 2016 at a single tertiary referral centre., Intervention: All men underwent 1.5-T mpMRI with an endorectal coil. All biopsies were performed by three operators as their first cases of targeted biopsy. Lesions with a Prostate Imaging Recording and Data System (PI-RADS) v.2 score of ≥3 detected at mpMRI were targeted. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: csPCa was defined as disease with a Gleason score at biopsy of ≥7. Operator expertise was coded as the progressive number of targeted biopsies performed by each physician. Multivariable logistic regression analyses (MVA) were used to assess the association between the targeted biopsy technique (FB vs CB) and operator expertise for detection of csPCa. Covariates consisted of prostate-specific antigen, prostate volume, PI-RADS v.2 (3 vs >3), number of targeted cores per MRI lesion, and digital rectal examination (negative vs positive). The same analyses were performed for patients undergoing FB only after accounting for the FB approach (transrectal vs transperineal). A lowess smoothing weighted function was used to graphically assess the effect of operator expertise on the probability of detecting csPCa, after accounting for all confounders., Results and Limitations: Overall, 157 patients (64%) underwent FB and 87 (36%) underwent CB. The overall csPCa detection rate was 58% for FB and 45% for CB (p=0.07). A significantly higher rate of csPCa detection in targeted samples was observed for FB compared to CB (57% vs 36%; p=0.002). On MVA, FB and operator expertise were significantly associated with a higher probability of csPCa detection in targeted samples (odds ratio [OR] 2.4 and 1.7, respectively; both p≤0.03). When the same analyses were repeated for patients undergoing FB, operator expertise remained an independent predictor of csPCa detection (OR 1.9; p=0.004). An increase in the probability of detecting csPCa with the number of procedures performed was observed after accounting for all confounders., Conclusions: We demonstrated that FB had higher detection rate than CB for csPCa. Moreover, operator expertise was significantly associated with higher detection rates for csPCa., Patient Summary: When different targeted biopsy techniques were compared, fusion biopsy provided a higher detection rate compared to cognitive biopsy for clinically significant prostate cancer (csPCa). Moreover, we found that operator expertise was an important predictor of the detection of csPCa, regardless of the procedure used., (Copyright © 2018 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

Muscles organise pseudo-crystalline arrays of actin, myosin and titin filaments to build force-producing sarcomeres. To study sarcomerogenesis, we have generated a transcriptomics resource of developing Drosophila flight muscles and identified 40 distinct expression profile clusters. Strikingly, most sarcomeric components group in two clusters, which are strongly induced after all myofibrils have been assembled, indicating a transcriptional transition during myofibrillogenesis. Following myofibril assembly, many short sarcomeres are added to each myofibril. Subsequently, all sarcomeres mature, reaching 1.5 µm diameter and 3.2 µm length and acquiring stretch-sensitivity. The efficient induction of the transcriptional transition during myofibrillogenesis, including the transcriptional boost of sarcomeric components, requires in part the transcriptional regulator Spalt major. As a consequence of Spalt knock-down, sarcomere maturation is defective and fibers fail to gain stretch-sensitivity. Together, this defines an ordered sarcomere morphogenesis process under precise transcriptional control - a concept that may also apply to vertebrate muscle or heart development., Competing Interests: MS, CB, AY, XZ, SL, AB, EB, GC, KB, BH, FS No competing interests declared, (© 2018, Spletter et al.)

Background: The ability to identify clinically significant prostate cancer (csPCa) has dramatically improved with the introduction of multiparametric magnetic resonance imaging (mpMRI). Given the growing interest in targeted biopsy and focal therapy, improving our knowledge on the relationship between mpMRI parameters and the ability to predict csPCa multifocality is mandatory., Objective: To assess whether the Prostate Imaging Reporting and Data System (PI-RADS) score for the index lesion (IL) may predict multifocal csPCa undetected by mpMRI., Design, Setting, and Participants: The study included 343 patients who underwent mpMRI of the prostate with subsequent biopsy between 2014 and 2017 at a single tertiary care referral centre., Intervention: Lesions with a PI-RADS v.2 score ≥2 detected at mpMRI (IL) were targeted with a fusion biopsy (Bx) approach (mpMRI-Bx). Moreover, each patient underwent a random extended transrectal ultrasound-guided biopsy (TRUS-Bx) during the same session. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: csPCa outside the IL was defined as disease detected at TRUS-Bx with a Gleason score (GS)≥3+4 and equal to or greater than the GS for the IL. The extent of csPCa detected in target and random cores was reported and stratified according to the GS and PI-RADS score for the IL. The probability of diagnosing csPCa outside the IL according to the PI-RADS score was also assessed in multivariable logistic regression analyses (MVA) after accounting for confounders., Results and Limitations: The detection rate for csPCa outside the IL was 30%. The detection rate for csPCa at TRUS-Bx was 8% for PI-RADS 2, 15% for PI-RADS 3, 36% for PI-RADS 4, and 58% for PI-RADS 5 lesions (p=0.03). Overall, the median length of csPCa found at TRUS-Bx and thus missed at mpMRI was 2.6mm. However, the length significantly increased with PI-RADS score for the IL, and was 1.8, 2.3, 2.8, and 3.8mm for PI-RADS 2, 3, 4, and 5 lesions, respectively (p=0.03). On MVA, PI-RADS 4 (odds ratio [OR] 7.6; p=0.008) and PI-RADS 5 scores (OR 17.3; p<0.001) were independent predictors of the presence of csPCa outside the IL. The study is limited by its retrospective design., Conclusions: Overall, the accuracy of mpMRI in identifying multifocal csPCa is poor, missing low-volume csPCa in approximately 30% of patients. Moreover, the rate and the extent of csPCa undetected by mpMRI significantly increased with the PI-RADS score for the IL, which can thus be considered a proxy for tumour multifocality., Patient Summary: The accuracy of multiparametric magnetic resonance imaging in identifying prostate cancer multifocality is poor. False negative findings were highly related to the PI-RADS score of the index lesion. These findings raise concerns about the indication for targeting the index lesion only when considering prostate biopsy and focal approaches., (Copyright © 2018 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

Genome ejection proteins are required to facilitate transport of bacteriophage P22 double-stranded DNA safely through membranes of Salmonella. The structures and locations of all proteins in the context of the mature virion are known, with the exception of three ejection proteins. Furthermore, the changes that occur to the proteins residing in the mature virion upon DNA release are not fully understood. We used cryogenic electron microscopy to obtain what is, to our knowledge, the first asymmetric reconstruction of mature bacteriophage P22 after double-stranded DNA has been extruded from the capsid-a state representative of one step during viral infection. Results of icosahedral and asymmetric reconstructions at estimated resolutions of 7.8 and 12.5 Å resolutions, respectively, are presented. The reconstruction shows tube-like protein density extending from the center of the tail assembly. The portal protein does not revert to the more contracted, procapsid state, but instead maintains an extended and splayed barrel structure. These structural details contribute to our understanding of the molecular mechanism of P22 phage infection and also set the foundation for future exploitation serving engineering purposes., (Copyright © 2018 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

The combined effects of grain germination and of subsequent fermentation on the physicochemical properties of sorghum flour were investigated by studying the structural changes occurring in the starch and protein fractions and by assessing their effects on physical properties of the resulting materials most relevant to end use. The sequential treatments were more effective than either individual treatment in the modification of starch-related properties, whereas modification of the protein components only occurs in the fermentation step, almost regardless of a previous germination step. The resulting profile of physicochemical traits offers several hints as for the suitability of flour from treated sorghum as an ingredient for various types of gluten-free food products, and provides a basis for expanding the use of processed sorghum in applications other than traditional African foods.

Unlabelled: The adeno-associated viruses (AAV) are promising therapeutic gene delivery vectors and better understanding of their capsid assembly and genome packaging mechanism is needed for improved vector production. Empty AAV capsids assemble in the nucleus prior to genome packaging by virally encoded Rep proteins. To elucidate the capsid determinants of this process, structural differences between wild-type (wt) AAV2 and a packaging deficient variant, AAV2-R432A, were examined using cryo-electron microscopy and three-dimensional image reconstruction both at an ∼5.0-Å resolution (medium) and also at 3.8- and 3.7-Å resolutions (high), respectively. The high resolution structures showed that removal of the arginine side chain in AAV2-R432A eliminated hydrogen bonding interactions, resulting in altered intramolecular and intermolecular interactions propagated from under the 3-fold axis toward the 5-fold channel. Consistent with these observations, differential scanning calorimetry showed an ∼10°C decrease in thermal stability for AAV2-R432A compared to wt-AAV2. In addition, the medium resolution structures revealed differences in the juxtaposition of the less ordered, N-terminal region of their capsid proteins, VP1/2/3. A structural rearrangement in AAV2-R432A repositioned the βA strand region under the icosahedral 2-fold axis rather than antiparallel to the βB strand, eliminating many intramolecular interactions. Thus, a single amino acid substitution can significantly alter the AAV capsid integrity to the extent of reducing its stability and possibly rendering it unable to tolerate the stress of genome packaging. Furthermore, the data show that the 2-, 3-, and 5-fold regions of the capsid contributed to producing the packaging defect and highlight a tight connection between the entire capsid in maintaining packaging efficiency., Importance: The mechanism of AAV genome packaging is still poorly understood, particularly with respect to the capsid determinants of the required capsid-Rep interaction. Understanding this mechanism may aid in the improvement of AAV packaging efficiency, which is currently ∼1:10 (10%) genome packaged to empty capsid in vector preparations. This report identifies regions of the AAV capsid that play roles in genome packaging and that may be important for Rep recognition. It also demonstrates the need to maintain capsid stability for the success of this process. This information is important for efforts to improve AAV genome packaging and will also inform the engineering of AAV capsid variants for improved tropism, specific tissue targeting, and host antibody escape by defining amino acids that cannot be altered without detriment to infectious vector production., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

The clinical suspicion of local recurrence of prostate cancer after radical treatment is based on the onset of biochemical failure. The use of multiparametric magnetic resonance imaging (MRI) for prostate cancer has increased over recent years, mainly for detection, staging, and active surveillance. However, suspicion of recurrence in the set of biochemical failure is becoming a significant reason for clinicians to request multiparametric MRI. Radiologists should be able to recognize the normal posttreatment MRI findings. Fibrosis and atrophic remnant seminal vesicles (SV) after radical prostatectomy are often found and must be differentiated from local relapse. Moreover, brachytherapy, external beam radiotherapy, and focal therapies tend to diffusely decrease the signal intensity of the peripheral zone on T2-weighted images due to the loss of water content, consequently mimicking tumor and hemorrhage. The combination of T2-weighted images and functional studies like diffusion-weighted imaging and dynamic contrast-enhanced imaging improves the identification of local relapse. Tumor recurrence tends to restrict on diffusion images and avidly enhances after contrast administration. The authors provide a review of the normal findings and the signs of local tumor relapse after radical prostatectomy, external beam radiotherapy, brachytherapy and focal therapies., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Objectives: The aim of this study is to develop a nomogram of clinical utility based on apparent diffusion coefficient (ADC) from diffusion-weighted imaging to predict extracapsular extension (ECE), and to validate externally its clinical utility., Materials and Methods: A total of 101 men (70 for the creation and 31 for external validation of the nomogram) underwent 1.5T multiparametric magnetic resonance imaging followed by radical prostatectomy at 2 different institutions. ADC values were assessed for normal and pathological tissue. Clinical and pathological variables were investigated by univariate and multivariate logistic regression analyses on 70 patients and logistic regression coefficients were used to develop our nomogram. Receiver operating characteristic curve analysis was performed to determine the optimal ADC cut off for ECE. The nomogram was then externally validated on 31 patients at another institution., Results: At univariate analysis, the following variables were associated with ECE: pathological ADC and Gleason at biopsy (P<0.001) along with tumor volume and ECE at imaging (P = 0.003). At multivariate analysis, pathological ADC (P = 0.027), tumor volume (P = 0.011), and biopsy Gleason (P = 0.040) maintained their independent predictor status and were included in our nomogram together with normal ADC and ECE at imaging. Our nomogram showed a significant higher sensitivity (88%) than T2-weighted imaging (54%; P = 0.010). External validation resulted in an overall accuracy of 81%., Conclusions: ADC represents a potential imaging biomarker to predict side-specific ECE in patients with prostate cancer. Our nomogram could improve the current diagnostic pathway and possibly the therapeutic approach for this disease., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Cervical cancer is the second most prevalent malignant tumor among women worldwide. High-risk human papillomaviruses (HPVs) are believed to be the major causative pathogens of mucosal epithelial cancers including cervical cancer. The HPV capsid is made up of 360 copies of major (L1) and 72 copies of minor (L2) capsid proteins. To date, limited high-resolution structural information about the HPV capsid has hindered attempts to understand details concerning the mechanisms by which HPV assembles and infects cells. In this study, we have constructed a pseudo-atomic model of the HPV59 L1-only capsid and demonstrate that the C-terminal arm of L1 participates in virus-host interactions. Moreover, when conjugated to a scaffold protein, keyhole limpet hemocyanin (KLH), this arm is immunogenic in vivo. These results provide new insights that will help elucidate HPV biology, and hence pave a way for the design of next-generation HPV vaccines., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Two-dimensional (2D) crystalline materials possess unique structural, mechanical and electronic properties that make them highly attractive in many applications. Although there have been advances in preparing 2D materials that consist of one or a few atomic or molecular layers, bottom-up assembly of 2D crystalline materials remains a challenge and an active area of development. More challenging is the design of dynamic 2D lattices that can undergo large-scale motions without loss of crystallinity. Dynamic behaviour in porous three-dimensional (3D) crystalline solids has been exploited for stimuli-responsive functions and adaptive behaviour. As in such 3D materials, integrating flexibility and adaptiveness into crystalline 2D lattices would greatly broaden the functional scope of 2D materials. Here we report the self-assembly of unsupported, 2D protein lattices with precise spatial arrangements and patterns using a readily accessible design strategy. Three single- or double-point mutants of the C4-symmetric protein RhuA were designed to assemble via different modes of intermolecular interactions (single-disulfide, double-disulfide and metal-coordination) into crystalline 2D arrays. Owing to the flexibility of the single-disulfide interactions, the lattices of one of the variants ((C98)RhuA) are essentially defect-free and undergo substantial, but fully correlated, changes in molecular arrangement, yielding coherently dynamic 2D molecular lattices. (C98)RhuA lattices display a Poisson's ratio of -1-the lowest thermodynamically possible value for an isotropic material-making them auxetic.

Unlabelled: Giardia lamblia virus (GLV) is a small, nonenveloped, nonsegmented double-stranded RNA (dsRNA) virus infecting Giardia lamblia, the most common protozoan pathogen of the human intestine and a major agent of waterborne diarrheal disease worldwide. GLV (genus Giardiavirus) is a member of family Totiviridae, along with several other groups of protozoal or fungal viruses, including Leishmania RNA viruses and Trichomonas vaginalis viruses. Interestingly, GLV is more closely related than other Totiviridae members to a group of recently discovered metazoan viruses that includes penaeid shrimp infectious myonecrosis virus (IMNV). Moreover, GLV is the only known protozoal dsRNA virus that can transmit efficiently by extracellular means, also like IMNV. In this study, we used transmission electron cryomicroscopy and icosahedral image reconstruction to examine the GLV virion at an estimated resolution of 6.0 Å. Its outermost diameter is 485 Å, making it the largest totivirus capsid analyzed to date. Structural comparisons of GLV and other totiviruses highlighted a related "T=2" capsid organization and a conserved helix-rich fold in the capsid subunits. In agreement with its unique capacity as a protozoal dsRNA virus to survive and transmit through extracellular environments, GLV was found to be more thermoresistant than Trichomonas vaginalis virus 1, but no specific protein machinery to mediate cell entry, such as the fiber complexes in IMNV, could be localized. These and other structural and biochemical findings provide a basis for future work to dissect the cell entry mechanism of GLV into a "primitive" (early-branching) eukaryotic host and an important enteric pathogen of humans., Importance: Numerous pathogenic bacteria, including Corynebacterium diphtheriae, Salmonella enterica, and Vibrio cholerae, are infected with lysogenic bacteriophages that contribute significantly to bacterial virulence. In line with this phenomenon, several pathogenic protozoa, including Giardia lamblia, Leishmania species, and Trichomonas vaginalis are persistently infected with dsRNA viruses, and growing evidence indicates that at least some of these protozoal viruses can likewise enhance the pathogenicity of their hosts. Understanding of these protozoal viruses, however, lags far behind that of many bacteriophages. Here, we investigated the dsRNA virus that infects the widespread enteric parasite Giardia lamblia. Using electron cryomicroscopy and icosahedral image reconstruction, we determined the virion structure of Giardia lamblia virus, obtaining new information relating to its assembly, stability, functions in cell entry and transcription, and similarities and differences with other dsRNA viruses. The results of our study set the stage for further mechanistic work on the roles of these viruses in protozoal virulence., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Unlabelled: As they mature, many capsids undergo massive conformational changes that transform their stability, reactivity, and capacity for DNA. In some cases, maturation proceeds via one or more intermediate states. These structures represent local minima in a rich energy landscape that combines contributions from subunit folding, association of subunits into capsomers, and intercapsomer interactions. We have used scanning calorimetry and cryo-electron microscopy to explore the range of capsid conformations accessible to bacteriophage HK97. To separate conformational effects from those associated with covalent cross-linking (a stabilization mechanism of HK97), a cross-link-incompetent mutant was used. The mature capsid Head I undergoes an endothermic phase transition at 60°C in which it shrinks by 7%, primarily through changes in its hexamer conformation. The transition is reversible, with a half-life of ~3 min; however, >50% of reverted capsids are severely distorted or ruptured. This observation implies that such damage is a potential hazard of large-scale structural changes such as those involved in maturation. Assuming that the risk is lower for smaller changes, this suggests a rationalization for the existence of metastable intermediates: that they serve as stepping stones that preserve capsid integrity as it switches between the radically different conformations of its precursor and mature states., Importance: Large-scale conformational changes are widespread in virus maturation and infection processes. These changes are accompanied by the release of conformational free energy as the virion (or fusogenic glycoprotein) switches from a precursor state to its mature state. Each state corresponds to a local minimum in an energy landscape. The conformational changes in capsid maturation are so radical that the question arises of how maturing capsids avoid being torn apart. Offering proof of principle, severe damage is inflicted when a bacteriophage HK97 capsid reverts from the (nonphysiological) state that it enters when heated past 60 °C. We suggest that capsid proteins have been selected in part by the criterion of being able to avoid sustaining collateral damage as they mature. One way of achieving this---as with the HK97 capsid-involves breaking the overall transition down into several smaller steps in which the risk of damage is reduced., (Copyright © 2014 Cardone et al.)

This paper reviews the most dependable heat flux sensors, which can be used with InfraRed (IR) thermography to measure convective heat transfer coefficient distributions, and some of their applications performed by the authors' research group at the University of Naples Federico II. After recalling the basic principles that make IR thermography work, the various heat flux sensors to be used with it are presented and discussed, describing their capability to investigate complex thermo-fluid-dynamic flows. Several applications to streams, which range from natural convection to hypersonic flows, are also described.

CUS-3 is a short-tailed, dsDNA bacteriophage that infects serotype K1 Escherichia coli. We report icosahedrally averaged and asymmetric, three-dimensional, cryo-electron microscopic reconstructions of the CUS-3 virion. Its coat protein structure adopts the "HK97-fold" shared by other tailed phages and is quite similar to that in phages P22 and Sf6 despite only weak amino acid sequence similarity. In addition, these coat proteins share a unique extra external domain ("I-domain"), suggesting that the group of P22-like phages has evolved over a very long time period without acquiring a new coat protein gene from another phage group. On the other hand, the morphology of the CUS-3 tailspike differs significantly from that of P22 or Sf6, but is similar to the tailspike of phage K1F, a member of the extremely distantly related T7 group of phages. We conclude that CUS-3 obtained its tailspike gene from a distantly related phage quite recently., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Papillomaviruses are a family of nonenveloped DNA viruses that infect the skin or mucosa of their vertebrate hosts. The viral life cycle is closely tied to the differentiation of infected keratinocytes. Papillomavirus virions are released into the environment through a process known as desquamation, in which keratinocytes lose structural integrity prior to being shed from the surface of the skin. During this process, virions are exposed to an increasingly oxidative environment, leading to their stabilization through the formation of disulfide cross-links between neighboring molecules of the major capsid protein, L1. We used time-lapse cryo-electron microscopy and image analysis to study the maturation of HPV16 capsids assembled in mammalian cells and exposed to an oxidizing environment after cell lysis. Initially, the virion is a loosely connected procapsid that, under in vitro conditions, condenses over several hours into the more familiar 60-nm-diameter papillomavirus capsid. In this process, the procapsid shrinks by ~5% in diameter, its pentameric capsomers change in structure (most markedly in the axial region), and the interaction surfaces between adjacent capsomers are consolidated. A C175S mutant that cannot achieve normal inter-L1 disulfide cross-links shows maturation-related shrinkage but does not achieve the fully condensed 60-nm form. Pseudoatomic modeling based on a 9-Å resolution reconstruction of fully mature capsids revealed C-terminal disulfide-stabilized "suspended bridges" that form intercapsomeric cross-links. The data suggest a model in which procapsids exist in a range of dynamic intermediates that can be locked into increasingly mature configurations by disulfide cross-linking, possibly through a Brownian ratchet mechanism. Importance: Human papillomaviruses (HPVs) cause nearly all cases of cervical cancer, a major fraction of cancers of the penis, vagina/vulva, anus, and tonsils, and genital and nongenital warts. HPV types associated with a high risk of cancer, such as HPV16, are generally transmitted via sexual contact. The nonenveloped virion of HPVs shows a high degree of stability, allowing the virus to persist in an infectious form in environmental fomites. In this study, we used cryo-electron microscopy to elucidate the structure of the HPV16 capsid at different stages of maturation. The fully mature capsid adopts a rigid, highly regular structure stabilized by intermolecular disulfide bonds. The availability of a pseudoatomic model of the fully mature HPV16 virion should help guide understanding of antibody responses elicited by HPV capsid-based vaccines., (Copyright © 2014 Cardone et al.)

In cryo-electron microscopy and single particle analysis, data acquisition and image processing are generally carried out in sequential steps and computation of a three-dimensional reconstruction only begins once all the micrographs have been acquired. We are developing an integrated system for processing images of icosahedral particles during microscopy to provide reconstructed density maps in real-time at the highest possible resolution. The system is designed as a combination of pipelines to run in parallel on a computer cluster and analyzes micrographs as they are acquired, handling automatically all the processing steps from defocus estimation and particle picking to origin/orientation determination. An ab initio model is determined independently from the first micrographs collected, and new models are generated as more particles become available. As a proof of concept, we simulated data acquisition sessions using three sets of micrographs of good to excellent quality that were previously recorded from different icosahedral viruses. Results show that the processing of single micrographs can keep pace with an acquisition rate of about two images per minute. The reconstructed density map improves steadily during the image acquisition phase and its quality at the end of data collection is only moderately inferior to that obtained by expert users who processed semi-automatically all the micrographs after the acquisition. The current prototype demonstrates the advantages of integrating three-dimensional image processing with microscopy, which include an ability to monitor acquisition in terms of the final structure and to predict how much data and microscope resources are needed to achieve a desired resolution., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Despite being essential for successful infection, the molecular cues involved in host recognition and genome transfer of viruses are not completely understood. Bacterial outer membrane proteins A and C co-purify in lipid vesicles with bacteriophage Sf6, implicating both outer membrane proteins as potential host receptors. We determined that outer membrane proteins A and C mediate Sf6 infection by dramatically increasing its rate and efficiency. We performed a combination of in vivo studies with three omp null mutants of Shigella flexneri, including classic phage plaque assays and time-lapse fluorescence microscopy to monitor genome ejection at the single virion level. Cryo-electron tomography of phage 'infecting' outer membrane vesicles shows the tail needle contacting and indenting the outer membrane. Lastly, in vitro ejection studies reveal that lipopolysaccharide and outer membrane proteins are both required for Sf6 genome release. We conclude that Sf6 phage entry utilizes either outer membrane proteins A or C, with outer membrane protein A being the preferred receptor., (© 2014 John Wiley & Sons Ltd.)

The resolution of density maps from single particle analysis is usually measured in terms of the highest spatial frequency to which consistent information has been obtained. This calculation represents an average over the entire reconstructed volume. In practice, however, substantial local variations in resolution may occur, either from intrinsic properties of the specimen or for technical reasons such as a non-isotropic distribution of viewing orientations. To address this issue, we propose the use of a space-frequency representation, the short-space Fourier transform, to assess the quality of a density map, voxel-by-voxel, i.e. by local resolution mapping. In this approach, the experimental volume is divided into small subvolumes and the resolution determined for each of them. It is illustrated in applications both to model data and to experimental density maps. Regions with lower-than-average resolution may be mobile components or ones with incomplete occupancy or result from multiple conformational states. To improve the interpretability of reconstructions, we propose an adaptive filtering approach that reconciles the resolution to which individual features are calculated with the results of the local resolution map., (Published by Elsevier Inc.)

Single particle analysis is a valuable tool in cryo-electron microscopy for determining the structure of biological complexes. However, the conformational state and the preparation of the sample are factors that play a critical role in the ultimate attainable resolution. In some cases extensive analysis at the microscope of a sample under different conditions is required to derive the optimal acquisition conditions. Currently this analysis is limited to raw micrographs, thus conveying only limited information on the structure of the complex. We are developing a computing system that generates a three-dimensional reconstruction from a single micrograph acquired under cryogenic and low dose conditions, and containing particles with icosahedral symmetry. The system provides the microscopist with immediate structural information from a sample while it is in the microscope and during the preliminary acquisition stage. The system is designed to run without user intervention on a multi-processor computing resource and integrates all the processing steps required for the analysis. Tests performed on experimental data sets show that the probability of obtaining a reliable reconstruction from one micrograph is primarily determined by the quality of the sample, with success rates close to 100% when sample conditions are optimal, and decreasing to about 60% when conditions are sub-optimal. The time required to generate a reconstruction depends significantly on the diameter of the particles, and in most instances takes about 1min. The proposed approach can provide valuable three-dimensional information, albeit at low resolution, on conformational states, epitope binding, and stoichiometry of icosahedral multi-protein complexes., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Objective: To assess the complication rates and quality of life in patients eligible for focal therapy who underwent template-assisted transperineal prostate biopsy (TTPB)., Materials and Methods: Eighty-seven patients with low-risk prostate cancer (clinical stage T1c-T2a, prostate-specific antigen level ≤10 ng/mL, biopsy Gleason score ≤6), who were candidates for focal therapy, underwent TTPB. The study details are available from http://clinicaltrials.gov (NCT00928603). The primary outcomes were the complication rates, according to the Clavien-Dindo classification, and changes in the quality of life, evaluated using the International Prostate Symptom Score, International Index of Erectile Function, and Functional Assessment of Cancer Therapy-Prostate questionnaires, before and 1 month after TTPB., Results: The median patient age was 63.9 years (range 46-78), with a median Charlson comorbidity index of 2.2 (range 0-4). No statistically significant differences were observed when comparing the general and/or specific domains of the International Prostate Symptom Score, International Index of Erectile Function, and Functional Assessment of Cancer Therapy-Prostate results before and 1 month after TTPB (P >.05 for all). Using the Clavien-Dindo classification, we observed 37 cases of grade 1 complications, including 5 (6.1%) cases of macrohematuria, 13 (16%) of hemospermia, 11 (13.5%) of perineal hematoma, 3 (3.7%) of perineal hematoma and hemospermia, and 5 (6.1%) of macrohematuria and hemospermia. Three patients (3.7%) developed a grade II complication (ie, acute urinary retention). Prostate cancer was detected in 54 patients (62.1%). Of 57 patients, 16 (29.6%) were upgraded from Gleason score 3+3/atypical small acinar proliferation to Gleason score 7. Of the 54 patients with positive TTPB findings, 18 (25.3%) showed an anatomic correspondence between the results of previous biopsies and TTPB., Conclusion: TTPB did not appear to have a significant effect on the quality of life of candidates for focal therapy, and the Clavien-Dindo complication rate was negligible., (Copyright © 2013 Elsevier Inc. All rights reserved.)