Exercise training decreases oxidative stress in skeletal muscle of rats with pulmonary arterial hypertension.

The effects of exercise training on oxidative stress in gastrocnemius of rats with pulmonary hypertension were studied. Four groups were established: sedentary control (SC), sedentary monocrotaline (SM), trained control (TC), trained monocrotaline (TM). Exercise was applied for 4 weeks, 5 days/week, 50-60 min/session, at 60% of VO2 max. Right ventricular (RV) pressures were measured, heart and gastrocnemius were removed for morphometric/biochemical analysis. Lipid peroxidation (LPO), H2O2, GSH/GSSG, and activity/expression of antioxidant enzymes were evaluated. Increased RV hypertrophy, systolic and end-diastolic pressures (RVEDP) were observed in SM animals, and the RVEDP was decreased in TM vs. SM. H2O2, SOD-1, and LPO were higher in the SM group than in SC. In TM, H2O2 was further increased when compared to SM, with a rise in antioxidant defences and a decrease in LPO. GSH/GSSG was higher only in the TC group. Exercise induced an efficient antioxidant adaptation, preventing oxidative damage to lipids.

Factores de riesgo de enfermedad residual en la re-RTU en una gran cohorte de pacientes con enfermedad T1G3 / Risk factors for residual disease at re-TUR in a large cohort of T1G3 patients

Introducción y objetivos: Los objetivos de la resección transuretral (RTU) del tumor vesical son la resección completa de las lesiones y la realización de un diagnóstico correcto con el objetivo de estadificar adecuadamente al paciente. Es bien sabido que la presencia de músculo detrusor en el espécimen es un requisito previo para minimizar el riesgo de infraestadificación.La persistencia de enfermedad tras la resección de los tumores vesicales no es infrecuente, y es la razón por la que las guías europeas recomiendan una re-resección transuretral (re-RTU) para todos los tumores T1. Recientemente se ha publicado que, en los casos con inclusión de músculo en el espécimen, la re-RTU no afecta la progresión ni la supervivencia específica del cáncer.Presentamos aquí los factores relacionados con el paciente y el tumor que pueden influir en la presencia de enfermedad residual en la re-RTU.Material y métodosDe nuestra cohorte retrospectiva de 2.451 pacientes con tumores T1G3 primarios tratados inicialmente con bacilo de Calnette-Guérin (BCG), están disponibles los resultados patológicos de 934 pacientes (38,1%) que se sometieron a una re-RTU. El 74% tenía tumores multifocales, el 20% de los tumores tenía más de 3 cm de diámetro y el 26% tenía carcinoma in situ (CIS) concomitante. En este subgrupo de pacientes que se sometieron a una segunda RTU, no hubo enfermedad residual en 267 pacientes (29%) y se presentó enfermedad residual en 667 pacientes (71%): Ta en 378 (40%) y T1 en 289 (31%) pacientes. Se analizaron la edad, el sexo, el estado del tumor (primario/recurrente), la terapia intravesical previa, el tamaño del tumor, la multifocalidad del tumor, la presencia de CIS concomitante y la inclusión de músculo en el espécimen para evaluar los factores de riesgo de enfermedad residual en la re-RTU, tanto en los análisis univariantes, como en las regresiones logísticas multivariantes. (AU)

Introduction and objectives: The goals of transurethral resection of a bladder tumor (TUR) are to completely resect the lesions and to make a correct diagnosis in order to adequately stage the patient. It is well known that the presence of detrusor muscle in the specimen is a prerequisite to minimize the risk of under staging.Persistent disease after resection of bladder tumors is not uncommon and is the reason why the European Guidelines recommended a re-TUR for all T1 tumors. It was recently published that when there is muscle in the specimen, re-TUR does not influence progression or cancer specific survival.We present here the patient and tumor factors that may influence the presence of residual disease at re-TUR.Material and methodsIn our retrospective cohort of 2451 primary T1G3 patients initially treated with BCG, pathology results for 934 patients (38.1%) who underwent re-TUR are available. 74% had multifocal tumors, 20% of tumors were more than 3 cm in diameter and 26% had concomitant CIS.In this subgroup of patients who underwent re-TUR, there was no residual disease in 267 patients (29%) and residual disease in 667 patients (71%): Ta in 378 (40%) and T1 in 289 (31%) patients. Age, gender, tumor status (primary/recurrent), previous intravesical therapy, tumor size, tumor multi-focality, presence of concomitant CIS, and muscle in the specimen were analyzed in order to evaluate risk factors of residual disease at re-TUR, both in univariate analyses and multivariate logistic regressions. (AU)

Monitoring the haemodynamic response to visual stimulation in glaucoma patients.

In this paper, we used time-domain functional near infrared spectroscopy (TD-fNIRS) to evaluate the haemodynamic response function (HRF) in the occipital cortex following visual stimulation in glaucomatous eyes as compared to healthy eyes. A total of 98 subjects were enrolled in the study and clinically classified as healthy subjects, glaucoma patients (primary open-angle glaucoma) and mixed subjects (i.e. with a different classification for the two eyes). After quality check data were used from HRF of 73 healthy and 62 glaucomatous eyes. The amplitudes of the oxygenated and deoxygenated haemoglobin concentrations, together with their latencies with respect to the stimulus onset, were estimated by fitting their time course with a canonical HRF. Statistical analysis showed that the amplitudes of both haemodynamic parameters show a significant association with the pathology and a significant discriminating ability, while no significant result was found for latencies. Overall, our findings together with the ease of use and noninvasiveness of TD-NIRS, make this technique a promising candidate as a supporting tool for a better evaluation of the glaucoma pathology.

Risk factors for residual disease at re-TUR in a large cohort of T1G3 patients.

INTRODUCTION AND OBJECTIVES: The goals of transurethral resection of a bladder tumor (TUR) are to completely resect the lesions and to make a correct diagnosis in order to adequately stage the patient. It is well known that the presence of detrusor muscle in the specimen is a prerequisite to minimize the risk of under staging. Persistent disease after resection of bladder tumors is not uncommon and is the reason why the European Guidelines recommended a re-TUR for all T1 tumors. It was recently published that when there is muscle in the specimen, re-TUR does not influence progression or cancer specific survival. We present here the patient and tumor factors that may influence the presence of residual disease at re-TUR. MATERIAL AND METHODS: In our retrospective cohort of 2451 primary T1G3 patients initially treated with BCG, pathology results for 934 patients (38.1%) who underwent re-TUR are available. 74% had multifocal tumors, 20% of tumors were more than 3 cm in diameter and 26% had concomitant CIS. In this subgroup of patients who underwent re-TUR, there was no residual disease in 267 patients (29%) and residual disease in 667 patients (71%): Ta in 378 (40%) and T1 in 289 (31%) patients. Age, gender, tumor status (primary/recurrent), previous intravesical therapy, tumor size, tumor multi-focality, presence of concomitant CIS, and muscle in the specimen were analyzed in order to evaluate risk factors of residual disease at re-TUR, both in univariate analyses and multivariate logistic regressions. RESULTS: The following were not risk factors for residual disease: age, gender, tumor status and previous intravesical chemotherapy. The following were univariate risk factors for presence of residual disease: no muscle in TUR, multiple tumors, tumors > 3 cm, and presence of concomitant CIS. Due to the correlation between tumor multi-focality and tumor size, the multivariate model retained either the number of tumors or the tumor diameter (but not both), p < 0.001. The presence of muscle in the specimen was no longer significant, while the presence of CIS only remained significant in the model with tumor size, p < 0.001. CONCLUSIONS: The most significant factors for a higher risk of residual disease at re-TUR in T1G3 patients are multifocal tumors and tumors more than 3 cm. Patients with concomitant CIS and those without muscle in the specimen also have a higher risk of residual disease.

Reference Protocol to Assess Analytical Performance of Higher Order Structural Analysis Measurements: Results from an Interlaboratory Comparison.

Measurements of protein higher order structure (HOS) provide important information on stability, potency, efficacy, immunogenicity, and biosimilarity of biopharmaceuticals, with a significant number of techniques and methods available to perform these measurements. The comparison of the analytical performance of HOS methods and the standardization of the results is, however, not a trivial task, due to the lack of reference protocols and reference measurement procedures. Here, we developed a protocol to structurally alter and compare samples of somatropin, a recombinant biotherapeutic, and describe the results obtained by using a number of techniques, methods and in different laboratories. This, with the final aim to provide tools and generate a pool of data to compare and benchmark analytical platforms and define method sensitivity to structural changes. Changes in somatropin HOS, induced by the presence of zinc at increasing concentrations, were observed, both globally and at more localized resolution, across many of the methods utilized in this study and with different sensitivities, suggesting the suitability of the protocol to improve understanding of inter- and cross-platform measurement comparability and assess analytical performance as appropriate.

Influence of Laboratory Index on match performance. A comparison study to evaluate physical performance in professional soccer players of an Italian Elite Team.

OBJECTIVE: It was hypothesized that activity exercise at professional levels could lead to an increase in metabolic levels and a decrease in performance parameters. These trends are explained by physical activity as a cellular stressor. SUBJECTS AND METHODS: We used an algorithm, Laboratory Index which evaluates salivary cortisol, CK and d-ROMs, collected previously from elite Italian soccer players, compared to InStat Index. The last one estimates analytically the athlete's performance in soccer pitch, applying the Heath Maps. RESULTS: A good agreement between the two Index was obtained, especially for two players, who showed an ideal combined trend. CONCLUSIONS: We would investigate the clinical and activity profile of soccer players with the aim of providing information for the development of training strategies. Also, the performances, during training and match time, are an objective evaluation of the athlete's physical preparation. As a consequence, the combination of two Index could be used for a new approach to the sports world.

Hypoalbuminemia in COVID-19: assessing the hypothesis for underlying pulmonary capillary leakage.

BACKGROUND: Since the first observations of patients with COVID-19, significant hypoalbuminaemia was detected. Its causes have not been investigated yet. OBJECTIVE: We hypothesized that pulmonary capillary leakage affects the severity of respiratory failure, causing a shift of fluids and proteins through the epithelial-endothelial barrier. METHODS: One hundred seventy-four COVID-19 patients with respiratory symptoms, 92 admitted to the intermediate medicine ward (IMW) and 82 to the intensive care unit (ICU) at Luigi Sacco Hospital in Milan, were studied. RESULTS: Baseline characteristics at admission were considered. Proteins, interleukin 8 (IL-8) and interleukin 10 (IL-10) in bronchoalveolar lavage fluid (BALF) were analysed in 26 ICU patients. In addition, ten autopsy ultrastructural lung studies were performed in patients with COVID-19 and compared with postmortem findings in a control group (bacterial pneumonia-ARDS and H1N1-ARDS). ICU patients had lower serum albumin than IMW patients [20 (18-23) vs 28 (24-33) g L-1 , P < 0.001]. Serum albumin was lower in more compromised groups (lower PaO2 -to-FiO2 ratio and worst chest X-ray findings) and was associated with 30 days of probability of survival. Protein concentration was correlated with IL-8 and IL-10 levels in BALF. Electron microscopy examinations of eight out of ten COVID-19 lung tissues showed loosening of junctional complexes, quantitatively more pronounced than in controls, and direct viral infection of type 2 pneumocytes and endothelial cells. CONCLUSION: Hypoalbuminaemia may serve as severity marker of epithelial-endothelial damage in patients with COVID-19. There are clues that pulmonary capillary leak syndrome plays a key role in the pathogenesis of COVID-19 and might be a potential therapeutic target.

Feasibility and relevance of urine culture during stone fragmentation in patients undergoing percutaneous nephrolithotomy and retrograde intrarenal surgery: a prospective study.

PURPOSE: We evaluated if, during lithotripsy, bacteria may be detected in the irrigation fluid of percutaneous nephrolithotomy (PCNL) and retrograde intrarenal surgery (RIRS). The concordance between urine culture from stone fragmentation (SFUC), bladder (BUC), renal pelvic (RPUC) and stone (SC) was analyzed. We also assessed the correlation between variables and cultures and their association with systemic inflammatory response syndrome (SIRS) and of a positive SC. METHODS: We included 107 patients who underwent PCNL (n = 53) and RIRS (n = 54) from January 2017 to May 2018. Samples for RPUC were obtained by renal catheterization. Stone fragments and irrigation fluid sample were sent for culture. RESULTS: SFUC was positive in 17 (15.9%), BUC in 22 (20.6%), RPUC in 26 (24.3%) and SC in 30 patients (28%). The concordance between SFUC and SC was the highest among all cultures: 94.1%. SFUC and SC grew identical microorganisms in 15/17 (88.2%) patients. Out of 17 (15.9%) patients with SIRS, 8 (7.5%) had sepsis. SFUC had the highest PPV and specificity to detect positive SC and SIRS. Previous urinary tract infection, a preoperative nephrostomy, stone diameter and composition, staghorn calculi, PCNL, positive BUC, RPUC and SFUC were predictors of infected stone. Variables that indicate complex stones, complex PCNL and an infection of the upper tract were associated with SIRS. CONCLUSION: SFUC is technically feasible, easy to retrieve and to analyze. The spectrum of SFUC potential application in clinical practice is when is not possible to perform a SC, e.g. complete dusting or during micro-PCNL.

Does the administration of preoperative pembrolizumab lead to sustained remission post-cystectomy? First survival outcomes from the PURE-01 study☆.

BACKGROUND: Initial studies of preoperative checkpoint inhibition before radical cystectomy (RC) have shown promising pathologic complete responses. We aimed to analyze the survival outcomes of patients enrolled in the PURE-01 study (NCT02736266). PATIENTS AND METHODS: We report the results of the secondary end points of PURE-01 in the final population of 143 patients. In particular, we report the event-free survival (EFS) outcomes, defined as the time from the first cycle of pembrolizumab to radiographic disease progression precluding RC, initiation of neoadjuvant chemotherapy (NAC), recurrence after RC, or death from any cause. Other end points were recurrence-free survival (RFS) and overall survival (OS). Subgroup analyses were carried out, including pathological response category, clinical complete responses (CR) assessed via multiparametric magnetic resonance imaging (mpMRI), and molecular subtyping. Cox regression analyses for EFS were also carried out. RESULTS: After a median [interquartile range (IQR)] follow-up of 23 (15-29) months, 12- and 24-month EFS were 84.5% [95% confidence interval (CI): 78.5-90.9] and 71.7% (62.7-82). The prognosis was favorable across all the different pathological response subgroups, with the exception of ypN+ (N = 21), showing a 24-month RFS (95% CI) of 39.3% (19.2% to 80.5%). A statistically significant EFS benefit was observed in patients with a clinical CR (P = 0.002). Programmed cell-death-ligand-1 combined positive score was significantly associated with longer EFS in multivariable analyses. Four patients refused RC after clinical evidence of CR, and none of them have recurred after a median follow-up of 10 months (IQR: 11-15). The claudin-low subtype displayed a numerically longer EFS after pembrolizumab and RC compared with the other subtypes. CONCLUSIONS: The EFS results from PURE-01 revealed that the immunotherapy effect was maintained post-RC in most patients. Pembrolizumab compared favorably with neoadjuvant chemotherapy, irrespective of the biomarker status. Molecular subtyping may be a useful tool to select the patients who are predicted to benefit the most from neoadjuvant pembrolizumab.

Glacier algae foster ice-albedo feedback in the European Alps.

The melting of glaciers and ice sheets is nowadays considered a symbol of climate change. Many complex mechanisms are involved in the melting of ice, and, among these processes, surface darkening due to organic material on bare ice has recently received attention from the scientific community. The presence of microbes on glaciers has been shown to decrease the albedo of ice and promote melting. Despite several studies from the Himalaya, Greenland, Andes, and Alaska, no quantitative studies have yet been conducted in the European Alps. In this paper, we made use of DNA sequencing, microscopy and field spectroscopy to describe the nature of glacier algae found at a glacier (Vadret da Morteratsch) of the European Alps and to evaluate their effect on the ice-albedo feedback. Among different algal species identified in the samples, we found a remarkable abundance of Ancylonema nordenskioeldii, a species that has never previously been quantitatively documented in the Alps and that dominates algal blooms on the Greenland Ice Sheet. Our results show that, at the end of the ablation season, the concentration of Ancylonema nordenskioeldii on the glacier surface is higher than that of other algal species (i.e. Mesotaenium berggrenii). Using field spectroscopy data, we identified a significant correlation between a reflectance ratio (750 nm/650 nm) and the algae concentration. This reflectance ratio could be useful for future mapping of glacier algae from remote sensing data exploiting band 6 (740 nm) and band 4 (665 nm) of the MultiSpectral Instrument (MSI) on board Sentinel-2 satellite. Here we show that the biological darkening of glaciers (i.e. the bioalbedo feedback) is also occurring in the European Alps, and thus it is a global process that must be taken into account when considering the positive feedback mechanisms related to glacier melting.

ODF1, sperm flagelar protein is expressed in kidney collecting ducts of rats.

ODF1 has been described as an exclusively expressed testicular protein and is located in the outer dense fibers along the sperm tail. ODF1 has been involved in the sperm motility and in the development of the flagellum, but the function of ODF1 is not already clear. Other ODF proteins, such as ODF2 have been characterized in other tissues like the basal body of the kidney primary cilium, but so far only the mRNA of ODF1 has been described in other tissues. These observations let us to hypothesize that the expression of the protein ODF1 could not be limited to the testis. Therefore, in the present work we proposed to evaluate if the ODF1 protein could also be present in tissues other than the testis. Here we demonstrated through western blot, immunofluorescence, and RT-PCR techniques that the protein and mRNA of ODF1 have been identified in the rat kidney. Finally, the presence of ODF1 in kidney has also been confirmed through proteomic analysis using mass spectrometry. The results derived from these different complementary approaches indicate that, to our knowledge and for the first time, ODF1 is demonstrated to be present in an additional organ different to testis. This results raise new questions about potential other functions and locations of the ODF1 protein.

Development of a new high sensitivity mechanical switch for augmentative and alternative communication access in people with amyotrophic lateral sclerosis.

BACKGROUND: People with Amyotrophic Lateral Sclerosis (PwALS) in the advanced phase are critically affected by an almost total loss of mobility and severe communication problems. Scanning access based on the patient's interaction with a sensor (or switch) that intercepts even a weak body movement is a valid communication aid. However, its use becomes limited with the progressive decline of residual movements. To overcome this problem, we designed a new sensor, the Lever Magnetic-spring Mechanical Switch (LeMMS), allowing repeated activation/release cycles requiring a very small activation force. METHODS: The LeMMS was applied and validated in a group of 20 PwALS in an advanced stage of disease. All subjects were regular users of communication aids employing other sensors, but which they could no longer operate their sensors (different from LeMMS). Patients were assessed at baseline (t0) and after one (t1), 6 (t2) and 12 (t3) months. Assessment at t0 included administration of standardized clinical scales, the Click-Test-30 counting the maximum number of LeMMS activations in 30 s, and thumb/fingers strength assessment with the Kendall scale. The QUEST 2.0-Dev questionnaire was administered at t1. Some use-related information and the Click-Test-30 were collected at t1, t2 and t3. RESULTS: After one training session, all patients could operate the LeMMS with minimal residual movement of one finger. At t1, they used it on average 5.45 h/day. The mean score of the QUEST 2.0-Dev was 4.63, suggesting strong satisfaction with the LeMMS. Regarding Click-Test-30 scores, no significant difference was found between t0 and t1, but performance at t2 and t3 declined significantly (p < 0.005 vs. t0). At t3, 9/20 patients were still able to use their communication aid. CONCLUSIONS: This new switch sensor can enable PwALS to use their communication aids for a prolonged time even in the advanced phase of disease. It is easy to use, reliable and cheap, thus representing an intermediate alternative to more sophisticated and costly devices.

Methods of aril removal and lightness conditions on seeds physiological quality of sour passion fruits / Métodos de remoção do arilo e condições de luminosidade sobre a qualidade fisiológica das sementes de maracujá azedo

Abstract Passion fruits species presents a larger economic importance for in natura consumption and industry. The main way to propagate these species is by seeds; however, the aril presence and environmental conditions may inhibit the seeds germination. In this sense, the objective of this study was to evaluate the effect of aril removal methods and light conditions on P. edulis var. flavicarpa seeds quality and germination. There were used five methods to remove the seeds aril: (1) friction on steel mesh sieve for 5 minutes; (2) friction with coarse sand on steel mesh sieve for 5 minutes; (3) immersion in lime solution at 10% (w/v), for 30 minutes; (4) fermentation in sugar solution at 10% (w/v), during 48 hours; and (5) natural pulp fermentation, during five days. The seeds were submitted to germination test at darkness and lightness, as well as, to length seedlings and emergence test. Regardless the aril removal method, the germination percentage was lower at lightness than darkness highlighting the natural pulp fermentation treatment, which presented 86% of seeds germinated. On the other hand, the aril removal methods did not affect the seedlings emergence, with emergence percentages ranged from 88 to 94%.

Resumo As espécies de maracujá apresentam grande importância econômica, tanto para o consumo in natura, quanto para a indústria. A principal maneira de propagar essas espécies é por sementes; contudo, a presença de arilo e condições ambientais pode inibir a germinação. Nesse sentido, objetivou-se estudar o efeito de métodos de remoção do arilo e luminosidade na qualidade e germinação das sementes de Passiflora edulis var. flavicarpa. Cinco métodos foram empregados para a remoção do arilo: (1) fricção em peneira de malha de aço por cinco minutos; (2) fricção com areia grossa em peneira de malha de aço por cinco minutos; (3) imersão em solução de cal a 10% (m/v), por 30 minutos; (4) fermentação da polpa em solução de açúcar a 10% (m/v), por 48 horas; e (5) fermentação natural da polpa, por cinco dias. As sementes foram submetidas ao teste de germinação no escuro e no claro, teste de comprimento de plântulas e teste de emergência. Independente do método de remoção do arilo, a porcentagem de germinação no claro apresentou-se menor em comparação ao escuro, com destaque para o tratamento de fermentação natural da polpa, o qual apresentou 86% de sementes germinadas. Por outro lado, os métodos de remoção do arilo não afetaram a emergência das plântulas, com porcentagem de emergência variando entre 88 a 94%.

The SonicHand Protocol for Rehabilitation of Hand Motor Function: A Validation and Feasibility Study.

Musical sonification therapy is a new technique that can reinforce conventional rehabilitation treatments by increasing therapy intensity and engagement through challenging and motivating exercises. The aim of this paper is to evaluate the feasibility and validity of the SonicHand protocol, a new training and assessment method for the rehabilitation of hand function. The study was conducted in 15 healthy individuals and 15 stroke patients. The feasibility of implementation of the training protocol was tested in stroke patients only, who practiced a series of exercises concurrently to music sequences produced by specific movements. The assessment protocol evaluated hand motor performance during pronation/supination, wrist horizontal flexion/extension, and hand grasp without sonification. From hand position data, 15 quantitative parameters were computed evaluating mean velocity, movement smoothness, and angular excursions of hand/fingers. We validated this assessment in terms of its ability to discriminate between patients and healthy subjects, test-retest reliability and concurrent validity with the upper limb section of the Fugl-Meyer scale (FM), the functional independence measure (FIM), and the Box and Block Test (BBT). All patients showed a good understanding of the assigned tasks and were able to correctly execute the proposed training protocol, confirming its feasibility. A moderate-to-excellent intraclass correlation coefficient was found in 8/15 computed parameters. The moderate-to-strong correlation was found between the measured parameters and the clinical scales. The SonicHand training protocol is feasible and the assessment protocol showed good to excellent between-group discrimination ability, reliability, and concurrent validity, thus enabling the implementation of new personalized and motivating training programs employing sonification for the rehabilitation of hand function.

Methods of aril removal and lightness conditions on seeds physiological quality of sour passion fruits.

Passion fruits species presents a larger economic importance for in natura consumption and industry. The main way to propagate these species is by seeds; however, the aril presence and environmental conditions may inhibit the seeds germination. In this sense, the objective of this study was to evaluate the effect of aril removal methods and light conditions on P. edulis var. flavicarpa seeds quality and germination. There were used five methods to remove the seeds aril: (1) friction on steel mesh sieve for 5 minutes; (2) friction with coarse sand on steel mesh sieve for 5 minutes; (3) immersion in lime solution at 10% (w/v), for 30 minutes; (4) fermentation in sugar solution at 10% (w/v), during 48 hours; and (5) natural pulp fermentation, during five days. The seeds were submitted to germination test at darkness and lightness, as well as, to length seedlings and emergence test. Regardless the aril removal method, the germination percentage was lower at lightness than darkness highlighting the natural pulp fermentation treatment, which presented 86% of seeds germinated. On the other hand, the aril removal methods did not affect the seedlings emergence, with emergence percentages ranged from 88 to 94%.

Detection of arbuscular mycorrhizal fungi associated with pecan (Caryaillinoinensis) trees by molecular and morphological approaches.

Arbuscular mycorrhizal (AM) fungal community associated with pecan (Caryaillinoinensis) roots and rhizospheric soils was assessed by spore isolation and morphological characterisation and by pyrosequencing of AM molecular markers. The AM fungal community associated with pecan growing in the field, was always more diverse than that associated with pecan growing in containers. This was not observed when AM richness was studied, suggesting that soil disturbance by a reduction in host plant richness leads to a less equitable distribution of AM fungal species, in contrast to natural soils. The chosen primers (AMV4.5F/AMDGR) for pyrosequencing showed high AM fungal specificity. Based on 97% sequence similarity, 49 operational taxonomic units (MOTUs) were obtained and, amongst these, 41 MOTUs corresponded to the Glomeromycotaphylum. The number of obtained AM sequences ranged from 2164, associated with field samples, to 5572 obtained from pecan trap pot culture samples, defining 30 and 29 MOTUs, respectively. Richness estimated by conventional species identification was 6 and 9 AM fungal species in soil and pot samples, respectively. Claroideoglomuslamellosum, Funneliformismosseae and Entrophosporainfrequens were the only taxa detected using both techniques. Predominant sequences in the pecan rhizosphere samples, such as Rhizoglomusirregulare and other less abundant (Dominikiairanica, Dominikiaindica, Sclerocystissinuosa, Paraglomuslaccatum), were detected only by pyrosequencing. Detection of AM fungal species based on spore morphology, in combination with molecular approaches, provides a more comprehensive estimate of fungal community composition.

Predictors of oncological outcomes in T1G3 patients treated with BCG who undergo radical cystectomy.

PURPOSE: To evaluate the oncological impact of postponing radical cystectomy (RC) to allow further conservative therapies prior to progression in a large multicentre retrospective cohort of T1-HG/G3 patients initially treated with BCG. METHODS: According to the time of RC, the population was divided into 3 groups: patients who did not progress to muscle-invasive disease, patients who progressed before radical cystectomy and patients who experienced progression at the time of radical cystectomy. Clinical and pathological outcomes were compared across the three groups. RESULTS: Of 2451 patients, 509 (20.8%) underwent RC. Patients with tumors > 3 cm or with CIS had earlier cystectomies (HR = 1.79, p = 0.001 and HR = 1.53, p = 0.02, respectively). Patients with tumors > 3 cm, multiple tumors or CIS had earlier T3/T4 or N + cystectomies. In patients who progressed, the timing of cystectomy did not affect the risk of T3/T4 or N + disease at RC. Patients with T3/T4 or N + disease at RC had a shorter disease-specific survival (HR = 4.38, p < 0.001), as did patients with CIS at cystectomy (HR = 2.39, p < 0.001). Patients who progressed prior to cystectomy had a shorter disease-specific survival than patients for whom progression was only detected at cystectomy (HR = 0.58, p = 0.024) CONCLUSIONS: Patients treated with RC before experiencing progression to muscle-invasive disease harbor better oncological and survival outcomes compared to those who progressed before RC and to those upstaged at surgery. Tumor size and concomitant CIS at diagnosis are the main predictors of surgical treatment while tumor size, CIS and tumor multiplicity are associated with extravesical disease at surgery.

Recurrence, progression and cancer-specific mortality according to stage at re-TUR in T1G3 bladder cancer patients treated with BCG: not as bad as previously thought.

PURPOSE: The goals of transurethral resection of a bladder tumor (TUR) are to completely resect the lesions and to make a correct diagnosis to adequately stage and treat the patient. Persistent disease after TUR is not uncommon and is why re-TUR is recommended in T1G3 patients. When there is T1 tumor in the re-TUR specimen, very high risks of progression (82%) have been reported. We analyze the risks of recurrence, progression to muscle-invasive disease and cancer-specific mortality (CSM) according to tumor stage at re-TUR in T1G3 patients treated with BCG. METHODS: In our retrospective cohort of 2451 T1G3 patients, 934 patients (38.1%) underwent re-TUR. 667 patients had residual disease (71.4%): Ta in 378 (40.5%), T1 in 289 (30.9%) patients. Times to recurrence, progression and CSM in the three groups were estimated using cumulative incidence functions and compared using the Cox regression model. RESULTS: During a median follow-up of 5.2 years, 512 patients recurred. The recurrence rate was significantly higher in patients with a T1 at re-TUR (P < 0.001). Progression rates differed according to the pathology at re-TUR, 25.3% in T1, 14.6% in Ta and 14.2% in case of no residual tumor (P < 0.001). Similar trends were seen in both patients with and without muscle in the original TUR specimen. CONCLUSIONS: Patients with T1G3 tumors and no residual disease or Ta at re-TUR have better recurrence, progression and CSM rates than previously reported, with a CSM rate of 13.1 and a 25.3% progression rate in re-TUR T1 disease.