Ectomycorrhizal fungi and the nitrogen economy of Nothofagus in southern Patagonia.

Subantarctic Nothofagus forests are the southernmost forests in the world, with negligible atmospheric nitrogen (N) deposition. Most paradigms about the role of ectomycorrhizal (ECM) fungi in N cycling and plant N uptake at high latitudes have been tested in boreal coniferous forests, while in the southern hemisphere, ECM hosts are primarily angiosperms. Using ITS1 meta-barcoding, we characterized ECM and saprotrophic fungal communities in evergreen and deciduous Nothofagus forests forming monodominant and mixed stands in the archipelago of Tierra del Fuego (Chile and Argentina). We assessed the N economy of Nothofagus by correlating host species with fungal relative abundances, edaphic variables, net N mineralization, microbial biomass N and the activity of eight extracellular soil enzymes activities. The N economy of deciduous N. pumilio forests was strikingly similar to boreal coniferous forests, with the lowest inorganic N availability and net N mineralization, in correlation to higher relative abundances of ECM fungi with enzymatic capacity for organic N mobilization (genus Cortinarius). In contrast, the N economy of evergreen N. betuloides forests was predominantly inorganic and correlated with ECM lineages from the family Clavulinaceae, in acidic soils with poor drainage. Grassy understory vegetation in deciduous N. antarctica forests likely promoted saprotrophic fungi (i.e., genus Mortierella) in correlation with higher activities of carbon-degrading enzymes. Differences between Nothofagus hosts did not persist in mixed forests, illustrating the range of soil fertility of these ECM angiosperms and the underlying effects of soil and climate on Nothofagus distribution and N cycling in southern Patagonia.

Los bosques subantárticos de Nothofagus son los más australes del mundo, en donde la deposición atmosférica de nitrógeno (N) es casi nula. La mayoría de los estudios sobre el papel de hongos ectomicorrícicos (ECM) está basado en bosques de coníferas perennes, en el hemisferio norte; mientras que en el hemisferio sur los hospederos ECM son principalmente angiospermas. Caracterizamos las comunidades de hongos ECM y saprobios del suelo por secuenciación de ADN ambiental de la región ITS1 de 150 muestras recolectadas en bosques perennes, caducifolios y mixtos dominados por Nothofagus en el archipiélago de Tierra del Fuego (Argentina y Chile). Estudiamos la economía del N en diferentes bosques de Nothofagus, evaluando las correlaciones entre las comunidades de hongos ECM y saprobios con sus hospederos, variables edáficas, mineralización neta del N, biomasa microbiana del N y la actividad de ocho enzimas extracelulares. Encontramos que la economía del N de los bosques caducifolios de N. pumilio es sorprendentemente similar a la de los bosques de coníferas boreales, en los que encontramos los valores más bajos de N orgánico disponible y de mineralización neta del N, en correlación con mayor abundancia relativa de hongos ECM con capacidad enzimáticas de descomposición (género Cortinarius). En contraste, la economía del N de los bosques perennes de N. betuloides es predominantemente inorgánica y se correlaciona con abundancia alta de hongos ECM de la familia Clavulinaceae, preferentemente en suelos ácidos y anegadizos. La vegetación herbácea del sotobosque en los bosques caducifolios de N. antarctica posiblemente ha aumentado la abundancia de hongos saprobios (p. ej., Mortierella) que se correlaciona con una mayor actividad de enzimas de degradación del carbono. Las diferencias encontradas entre bosques perennes y caducifolios no persistieron en los bosques mixtos, lo que ilustra el gradiente de fertilidad del suelo de estas angiospermas formadoras de ectomicorrizas, y los efectos subyacentes del suelo y el clima en la distribución de Nothofagus y el ciclo del N en la región.

Comparative Study of Gleason 7 (3+4) and (4+3) Prostatic Adenocarcinomas with Prog-nostic Criteria and Immunohistochemical Profiles of AMACR, PSA and Ki-67.

BACKGROUND: To compare Gleason 7 (3+4) and (4+3) prostatic adenocarcinoma (PC) with different prognostic criteria through immunohistochemical analysis with anti-PSA, anti-Ki 67 and anti-AMARC antibodies. METHODS: We analyzed 221 surgical specimens from patients between 40 and 86 years-old (mean=63) with PC. The immunohistochemical study was performed with anti-PSA, anti-Ki 67 and anti-AMARC. The microscopic fields were photographed with an Olympus DP70 digital camera coupled to an Olympus BX51 microscope and archived in TIFF. Proportion and intensity criteria were used to quantify the anti-PSA antibody and for the anti-Ki 67 antibody, the quantification by similarity of this antibody in breast carcinomas. Anti-AMACR protein expression was based on four scores: negative, weak, moderate and strong. The statistical analysis was performed with the Graph Pad Prism 5 program. RESULTS: In the Gleason score 7 (3+4) we had 91.72% in pT2 and 8.27% in the pT3 group; 8.27% recurrences, of which 90.90% in the pT2 group. In the Gleason score 7 (4+3) we had 77.27% in the pT2 group and 22.72% in the pT3 group and 10.22% of relapses, of which 66.66% in the pT2 group and 33.33% in the pT3 group. In 6.81% of cases there was an increase in the anti-Ki 67 index and in 2.27% of the cases, there was an increase in the immunoexpression of anti-p53 when comparing Gleason score 7 (3+4) with Gleason score 7 (4+3). CONCLUSION: Our study confirmed differences in the Gleason score 7 (3+4) and Gleason score 7 (4+3) of PC when comparing prognostic criteria. Anti-Ki 67 and anti-PSA antibody immunostaining showed a positive correlation as the Gleason score 7 increased from (3+4) to (4+3).

Suitable Mouse Model to Study Dynamics of West Nile Virus Infection in Culex quinquefasciatus Mosquitoes.

West Nile Virus (WNV) poses a significant global public health threat as a mosquito-borne pathogen. While laboratory mouse models have historically played a crucial role in understanding virus biology, recent research has focused on utilizing immunocompromised models to study arboviruses like dengue and Zika viruses, particularly their interactions with Aedes aegypti mosquitoes. However, there has been a shortage of suitable mouse models for investigating WNV and St. Louis encephalitis virus interactions with their primary vectors, Culex spp. mosquitoes. Here, we establish the AG129 mouse (IFN α/ß/γ R-/-) as an effective vertebrate model for examining mosquito-WNV interactions. Following intraperitoneal injection, AG129 mice exhibited transient viremia lasting several days, peaking on the second or third day post-infection, which is sufficient to infect Culex quinquefasciatus mosquitoes during a blood meal. We also observed WNV replication in the midgut and dissemination to other tissues, including the fat body, in infected mosquitoes. Notably, infectious virions were present in the saliva of a viremic AG129 mouse 16 days post-exposure, indicating successful transmission capacity. These findings highlight the utility of AG129 mice for studying vector competence and WNV-mosquito interactions.

Prospective results of the minimally invasive laser enucleation of the prostate (MiLEP).

BACKGROUND: The objective of the present study is to prospectively analyze the prostate enucleation procedure with Holmium Laser using the minimally invasive technique (MiLEP), comparing the outcomes and their variables pre- and postoperatively. METHODS: We studied men aged 40 years or over, with prostate volumes greater than or equal to 35 cm³ with lower urinary tract symptoms due to BPH. We performed flowmetry and administered the IPSS questionnaire before and 6 months after the MiLEP procedure. The patients were operated on with a 60 W Holmium Laser (Cyber-Ho Quanta System®) using 54 W of power (energy 1.8 J and frequency of 30 Hz). Enucleation was performed using the en bloc technique with early sphincter release. After enucleation, the tissue was morcellated using a 22 Fr morcescope (RZ-Medizintechnik GmbH, Tuttlingen, Germany) and Piranha (Richard Wolf) morcellator. The final Hemostasis after morcelation was made using laser with 30wW power, energy at 1,0 joules and frequency at 30 Hertz. Student's T test and Man-Whitney was used to statistical analysis (p < 0.05). RESULTS: After selection we submitted 73 patients (mean age= 68.2 years) to MiLEP procedure with a follow up of 6 months. The prostate volume presented an average of 94.53 cm³ (65 to 112 cm³, SD = 5.363) preoperatively. The urinary continence rate after the procedure was greater than 95% after 1 week and 99% in the 1st month. All patients were continent after 6 months. The IPSS questionnaire before (mean = 21.18 points/SD = 6.557) and after (mean = 7.92 points/SD = 2.408) the MiLEP had statistical significance (p < 0.001). The flowmetry(ml/s) before (9.02/SD = 2.842) and after (21.07/SD = 6.228) the MiLEP had statistical significance (p < 0.001). The average time of the procedure was 78.5 min and the bladder catheter was removed after 18 h in mean. In 4 patients (5.8%) we observed hematuria and in 1 case (1.47%) the patient needs urinary catheterization. CONCLUSION: MiLEP is a safe and effective procedure, with significant improvement in urinary flow and symptoms in the short term. Although the results of this study were satisfactory and the urinary incontinence rate was lower compared to HoLEP data found in the literature, multicenter studies with longer follow-up are needed to confirm these findings.

A complete year of urology residency training under COVID-19: impact on education and health.

OBJECTIVES: To evaluate the impact of COVID-19 pandemics on clinical and surgical practice, educational activities, health and lifestyle behavior of Brazilian urology residents after 1 year of socio-economic restrictions. MATERIALS AND METHODS: An electronic survey was e-mailed to all postgraduate (PG) students registered by the Brazilian Society of Urology. The survey inclu-ded an assessment of socio-demographic, clinical practice, educational, health-related and behavior parameters. We also evaluated which subareas of urology were predominantly affected. A similar survey was adapted and sent to the directors of all urology residency programs. RESULTS: COVID-19 pandemic has severely impacted the clinical, surgical, and educational activities of urology residents in Brazil. Urology residents reported >50% decrease in multiple surgical modalities. We highlight kidney transplantation surgeries (66.2%), minor surgeries (62.3%), endoscopic surgeries (42.6%) and reconstructive surgeries (38.8%). This could represent a critical skills gap that residents may face beyond the COVID-19 pandemic. Furthermore, PG students faced stressful situations that caused worsening of mental and physical health, such as getting redirected to assistance of COVID-19 patients (66.9%), and high rate of infection by SARS-CoV-2 (58.2%). CONCLUSIONS: The COVID-19 pandemic has severely impacted the clinical, surgical, and educational activities of urology residents in Brazil. This could represent a critical skills gap that residents may face beyond the COVID-19 pandemic. PG students faced stressful situations that caused worsening of mental and physical health such as redirection to assistance of COVID-19 patients, concern about their own contamination and of family members.

High Variability in Standardized Outcome Thresholds of Clinically Important Changes in Shoulder Instability Surgery: A Systematic Review.

PURPOSE: To examine reported minimal clinically important difference (MCID) and patient-acceptable satisfactory state (PASS) values for patient-reported outcome measures (PROMs) after shoulder instability surgery and assess variability in published values depending on the surgery performed. Our secondary aims were to describe the methods used to derive MCID and PASS values in the published literature, including anchor-based, distribution-based, or other approaches, and to assess the frequency of MCID and PASS use in studies on shoulder instability surgery. METHODS: A systematic review of MCID and PASS values after Bankart, Latarjet, and Remplissage procedures was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA). The Embase, PubMed, and Cochrane Central Register of Controlled Trials (CENTRAL) databases were queried from 1985 to 2023. Inclusion criteria included studies written in English and studies reporting use of MCID or PASS for patient-reported outcome measures (PROMS) after Latarjet, Bankart, and Remplissage approaches for shoulder instability surgery. Extracted data included study population characteristics, intervention characteristics, and outcomes of interest. Continuous data were described using medians and ranges. Categorical variables, including PROMs and MCID/PASS methods, were described using percentages. Because MCID is a patient-level rather than a group-level metric, the authors confirmed that all included studies reported proportions (%) of subjects who met or exceeded the MCID. RESULTS: A total of 174 records were screened, and 8 studies were included in this review. MCID was the most widely used outcome threshold and was reported in all 8 studies, with only 2 studies reporting both the MCID and the PASS. The most widely studied PROMs were the American Shoulder and Elbow Surgeons (range 5.65-9.6 for distribution MCID, 8.5 anchor MCID, 86 anchor PASS); Single Assessment Numeric Evaluation (range 11.4-12.4 distribution MCID, 82.5-87.5 anchor PASS); visual analog scale (VAS) (range 1.1-1.7 distribution MCID, 1.5-2.5 PASS); Western Ontario Shoulder Instability Index (range 60.7-254.9 distribution MCID, 126.43 anchor MCID, 571-619.5 anchor PASS); and Rowe scores (range 5.6-8.4 distribution MCID, 9.7 anchor MCID). Notably, no studies reported on substantial clinical benefit or maximal outcome improvement. CONCLUSIONS: Despite the wide array of available PROMs for assessing shoulder instability surgery outcomes, the availability of clinically significant outcome thresholds such as MCID and PASS remains relatively limited. Although MCID has been the most frequently reported metric, there is considerable interstudy variability observed in their values. CLINICAL RELEVANCE: Knowing the outcome thresholds such as MCID and PASS of the PROMs frequently used to evaluate the results of glenohumeral stabilization surgery is fundamental because they allow us to know what is a clinically significant improvement for the patient.

A look beyond the QR code of SNARE proteins.

Soluble N-ethylmaleimide-sensitive factor Attachment protein REceptor (SNARE) proteins catalyze the fusion process of vesicles with target membranes in eukaryotic cells. To do this, they assemble in a zipper-like fashion into stable complexes between the membranes. Structural studies have shown that the complexes consist of four different helices, which we subdivide into Qa-, Qb-, Qc-, and R-helix on the basis of their sequence signatures. Using a combination of biochemistry, modeling and molecular dynamics, we investigated how the four different types are arranged in a complex. We found that there is a matching pattern in the core of the complex that dictates the position of the four fundamental SNARE types in the bundle, resulting in a QabcR complex. In the cell, several different cognate QabcR-SNARE complexes catalyze the different transport steps between the compartments of the endomembrane system. Each of these cognate QabcR complexes is compiled from a repertoire of about 20 SNARE subtypes. Our studies show that exchange within the four types is largely tolerated structurally, although some non-cognate exchanges lead to structural imbalances. This suggests that SNARE complexes have evolved for a catalytic mechanism, a mechanism that leaves little scope for selectivity beyond the QabcR rule.

Development of nerves and vessels in the penis during the human fetal period.

INTRODUCTION: Although nerves and vessels of the penis play important role in erection, there are few studies on their development in human fetus. Therefore, the objective of the present study is to analyze, quantitatively, in the corpora cavernosa and corpus spongiosum, the development of the nerves and vessels in the fetal penis at different gestational ages. MATERIAL AND METHODS: Fifty-six fresh, macroscopically normal human fetuses aged from 13 to 36 weeks post-conception (WPC) were used. Gestational age was determined by the foot length criterion. Penises were immediately fixed in 10% formalin, and routinely processed for paraffin embedding, after which tissue sections from the mid-shaft were obtained. We used immunohistochemical staining to analyze the nerves and vessels in the corpus cavernous and in the corpus spongiosum. These elements were identified and quantified as percentage by using the Image-J software. RESULTS: The quantitative analysis showed that the percentage of nerves varied from 3.03% to 20.35% in the corpora cavernosa and from 1.89% to 23.88% in the corpus spongiosum. The linear regression analysis indicated that nerves growth (incidence) in the corpora cavernosa and corpus spongiosum correlated significantly and positively with fetal age (r2=0.9421, p<0.0001) and (r2=0.9312, p<0.0001), respectively, during the whole fetal period studied. Also, the quantitative analysis showed that the percentage of vessels varies from 2.96% to 12.86% in the corpora cavernosa and from 3.62% to 14.85% in the corpus spongiosum. The linear regression analysis indicated that vessels growth (appearance) in the corpora cavernosa and corpus spongiosum correlated significantly and positively with fetal age (r2=0.8722, p<0.0001) and (r2=0.8218, p<0.0001), respectively, during the whole fetal period studied. In addition, the linear regression analysis demonstrated a more intense growth rate of nerves in the corpus spongiosum during the 2nd trimester of gestation, when compared with nerves in the corpora cavernosa. In addition, the linear regression analysis demonstrated a more intense growth rate of vessels in the corpus spongiosum when compared with the corpora cavernosa, during the whole fetal period studied. CONCLUSIONS: In the fetal period, the human penis undergoes major developmental changes, notably in the content and distribution of nerves and vessels. We found strong correlation between nerves and vessels growth (amount) with fetal age, both in the corpora cavernosa and corpus spongiosum. There is significant greater proportional number of nerves than vessels during the whole fetal period studied. Also, nerves and vessels grow in a more intense rate than that of the corpora cavernosa and corpus spongiosum areas.

Evidence of restorative therapies in the treatment of Peyronie disease: A narrative review.

OBJECTIVE: To describe the evidence of Platelet Rich Plasma (PRP), Stem cells therapy (SCT) and Extracorporeal shockwave therapy (ESWL) for the treatment of Peyronies disease (PD), including information from the main urological society guidelines. MATERIALS AND METHODS: A literature review of PubMed articles published between 2000 and 2023 was conducted, utilizing keywords such as "Peyronie's Disease", "Penile curvature", "Platelet Rich Plasma", "Stem cells", and "Extracorporeal shockwave therapy". Only full-text articles in English were included, excluding case reports and opinions. RESULTS: A considerable number of clinical trials were conducted using PRP penile injections for therapy of PD, showing reduction of curvature, plaque size and improvement in quality of life. Preclinical studies in rats have shown the potential benefit of adipose-derived stem cells, with improvements in erectile function and fibrosis. Human studies with mesenchymal stem cells demonstrated promising results, with reduction of curvature and plaque size. ESWL effects on PD were investigated in randomized clinical trials and demonstrated no significant impact in curvature or plaque size, but reasonable effect on pain control. CONCLUSION: Restorative therapies has emerged as an innovative treatment option for PD and the results from current studies appear to be promising and demonstrated good safety profile. Unfortunately, due to scarce evidence, PRP and SCT are still considered experimental by American Urological Association (AUA) and European Association of Urology (EAU) guidelines. ESWT is recommended, by the same guidelines, for pain control only. More high-quality studies with long-term follow-up outcomes are needed to evaluate efficacy and reproducibility of those therapies.

The DNA-PAINT palette: a comprehensive performance analysis of fluorescent dyes.

DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) is a super-resolution fluorescence microscopy technique that achieves single-molecule 'blinking' by transient DNA hybridization. Despite blinking kinetics being largely independent of fluorescent dye choice, the dye employed substantially affects measurement quality. Thus far, there has been no systematic overview of dye performance for DNA-PAINT. Here we defined four key parameters characterizing performance: brightness, signal-to-background ratio, DNA-PAINT docking site damage and off-target signal. We then analyzed 18 fluorescent dyes in three spectral regions and examined them both in DNA origami nanostructures, establishing a reference standard, and in a cellular environment, targeting the nuclear pore complex protein Nup96. Finally, having identified several well-performing dyes for each excitation wavelength, we conducted simultaneous three-color DNA-PAINT combined with Exchange-PAINT to image six protein targets in neurons at ~16 nm resolution in less than 2 h. We thus provide guidelines for DNA-PAINT dye selection and evaluation and an overview of performances of commonly used dyes.

Impact of nanosilver surface electronic distributions on serum protein interactions and hemocompatibility.

The translation of silver-based nanotechnology 'from bench to bedside' requires a deep understanding of the molecular aspects of its biological action, which remains controversial at low concentrations and non-spherical morphologies. Here, we present a hemocompatibility approach based on the effect of the distinctive electronic charge distribution in silver nanoparticles (nanosilver) on blood components. According to spectroscopic, volumetric, microscopic, dynamic light scattering measurements, pro-coagulant activity tests, and cellular inspection, we determine that at extremely low nanosilver concentrations (0.125-2.5µg ml-1), there is a relevant interaction effect on the serum albumin and red blood cells (RBCs). This explanation has its origin in the surface charge distribution of nanosilver particles and their electron-mediated energy transfer mechanism. Prism-shaped nanoparticles, with anisotropic charge distributions, act at the surface level, generating a compaction of the native protein molecule. In contrast, the spherical nanosilver particle, by exhibiting isotropic surface charge, generates a polar environment comparable to the solvent. Both morphologies induce aggregation at NPs/bovine serum albumin ≈ 0.044 molar ratio values without altering the coagulation cascade tests; however, the spherical-shaped nanosilver exerts a negative impact on RBCs. Overall, our results suggest that the electron distributions of nanosilver particles, even at extremely low concentrations, are a critical factor influencing the molecular structure of blood proteins' and RBCs' membranes. Isotropic forms of nanosilver should be considered with caution, as they are not always the least harmful.

A bacterial TIR-based immune system senses viral capsids to initiate defense.

Toll/interleukin-1 receptor (TIR) domains are present in immune systems that protect prokaryotes from viral (phage) attack. In response to infection, TIRs can produce a cyclic adenosine diphosphate-ribose (ADPR) signaling molecule, which activates an effector that depletes the host of the essential metabolite NAD+ to limit phage propagation. How bacterial TIRs recognize phage infection is not known. Here we describe the sensing mechanism for the staphylococcal Thoeris defense system, which consists of two TIR domain sensors, ThsB1 and ThsB2, and the effector ThsA. We show that the major capsid protein of phage Φ80α forms a complex with ThsB1 and ThsB2, which is sufficient for the synthesis of 1"-3' glycocyclic ADPR (gcADPR) and subsequent activation of NAD+ cleavage by ThsA. Consistent with this, phages that escape Thoeris immunity harbor mutations in the capsid that prevent complex formation. We show that capsid proteins from staphylococcal Siphoviridae belonging to the capsid serogroup B, but not A, are recognized by ThsB1/B2, a result that suggests that capsid recognition by Sau-Thoeris and other anti-phage defense systems may be an important evolutionary force behind the structural diversity of prokaryotic viruses. More broadly, since mammalian toll-like receptors harboring TIR domains can also recognize viral structural components to produce an inflammatory response against infection, our findings reveal a conserved mechanism for the activation of innate antiviral defense pathways.