The derlin Dfm1 couples retrotranslocation of a folded protein domain to its proteasomal degradation.

Endoplasmic reticulum (ER) proteins are degraded by proteasomes in the cytosol through ER-associated degradation (ERAD). This process involves the retrotranslocation of substrates across the ER membrane, their ubiquitination, and membrane extraction by the Cdc48/Npl4/Ufd1 ATPase complex prior to delivery to proteasomes for degradation. How the presence of a folded luminal domain affects substrate retrotranslocation and this event is coordinated with subsequent ERAD steps remains unknown. Here, using a model substrate with a folded luminal domain, we showed that Cdc48 ATPase activity is sufficient to drive substrate retrotranslocation independently of ERAD membrane components. However, the complete degradation of the folded luminal domain required substrate-tight coupling of retrotranslocation and proteasomal degradation, which was ensured by the derlin Dfm1. Mutations in Dfm1 intramembrane rhomboid-like or cytosolic Cdc48-binding regions resulted in partial degradation of the substrate with accumulation of its folded domain. Our study revealed Dfm1 as a critical regulator of Cdc48-driven retrotranslocation and highlights the importance of coordinating substrate retrotranslocation and degradation during ERAD.

Scanning Tunneling Spectroscopy Method for the Prediction of Semiconductor Heterojunction Performance as a Prequel for Device Development.

The prediction of semiconductor device performance is a persistent challenge in materials science, and the ability to anticipate useful specifications prior to construction is crucial for enhancing the overall efficiency. In this study, we investigate the constituents of a solar cell by employing scanning tunneling microscopy (STM) and spectroscopy (STS). Through our observations, we identify a spatial distribution of the dopant type in thin films of materials that were designed to present major p-doping for germanium sulfide (GeS) and dominant n-doping for tin disulfide (SnS2). By generating separate STS maps for each semiconductor film and conducting a statistical analysis of the gap and doping distribution, we determine intrinsic limitations for the solar cell efficiency that must be understood prior to processing. Subsequently, we fabricate a solar cell utilizing these materials (GeS and SnS2) via vapor phase deposition and carry out a characterization using standard J-V curves under both dark/illuminated irradiance conditions. Our devices corroborate the expected reduced efficiency due to doping fluctuation but exhibit stable photocurrent responses. As originally planned, quantum efficiency measurements reveal that the peak efficiency of our solar cell coincides with the range where the standard silicon solar cells sharply decline. Our STS method is suggested as a prequel to device development in novel material junctions or deposition processes where fluctuations of doping levels are retrieved due to intrinsic material characteristics such as the occurrence of defects, roughness, local chemical segregation, and faceting or step bunching.

Comparative Genomics and Characterization of SARS-CoV-2 P.1 (Gamma) Variant of Concern From Amazonas, Brazil.

BACKGROUND: P.1 lineage (Gamma) was first described in the State of Amazonas, northern Brazil, in the end of 2020, and has emerged as a very important variant of concern (VOC) of SARS-CoV-2 worldwide. P.1 has been linked to increased infectivity, higher mortality, and immune evasion, leading to reinfections and potentially reduced efficacy of vaccines and neutralizing antibodies. METHODS: The samples of 276 patients from the State of Amazonas were sent to a central referral laboratory for sequencing by gold standard techniques, through Illumina MiSeq platform. Both global and regional phylogenetic analyses of the successfully sequenced genomes were conducted through maximum likelihood method. Multiple alignments were obtained including previously obtained unique human SARS-CoV-2 sequences. The evolutionary histories of spike and non-structural proteins from ORF1a of northern genomes were described and their molecular evolution was analyzed for detection of positive (FUBAR, FEL, and MEME) and negative (FEL and SLAC) selective pressures. To further evaluate the possible pathways of evolution leading to the emergence of P.1, we performed specific analysis for copy-choice recombination events. A global phylogenomic analysis with subsampled P.1 and B.1.1.28 genomes was applied to evaluate the relationship among samples. RESULTS: Forty-four samples from the State of Amazonas were successfully sequenced and confirmed as P.1 (Gamma) lineage. In addition to previously described P.1 characteristic mutations, we find evidence of continuous diversification of SARS-CoV-2, as rare and previously unseen P.1 mutations were detected in spike and non-structural protein from ORF1a. No evidence of recombination was found. Several sites were demonstrated to be under positive and negative selection, with various mutations identified mostly in P.1 lineage. According to the Pango assignment, phylogenomic analyses indicate all samples as belonging to the P.1 lineage. CONCLUSION: P.1 has shown continuous evolution after its emergence. The lack of clear evidence for recombination and the positive selection demonstrated for several sites suggest that this lineage emergence resulted mainly from strong evolutionary forces and progressive accumulation of a favorable signature set of mutations.

Proxalutamide Reduces the Rate of Hospitalization for COVID-19 Male Outpatients: A Randomized Double-Blinded Placebo-Controlled Trial.

Antiandrogens have demonstrated a protective effect for COVOD-19 patients in observational and interventional studies. The goal of this study was to determine if proxalutamide, an androgen receptor antagonist, could be an effective treatment for men with COVID-19 in an outpatient setting. A randomized, double-blinded, placebo-controlled clinical trial was conducted at two outpatient centers (Brasilia, Brazil). Patients were recruited from October 21 to December 24, 2020 (clinicaltrials.gov number, NCT04446429). Male patients with confirmed COVID-19 but not requiring hospitalization (COVID-19 8-point ordinal scale <3) were administered proxalutamide 200 mg/day or placebo for up to 7 days. The primary endpoint was hospitalization rate at 30 days post-randomization. A total of 268 men were randomized in a 1:1 ratio. 134 patients receiving proxalutamide and 134 receiving placebo were included in the intention-to-treat analysis. The 30-day hospitalization rate was 2.2% in men taking proxalutamide compared to 26% in placebo, P < 0.001. The 30-day hospitalization risk ratio was 0.09; 95% confidence interval (CI) 0.03-0.27. Patients in the proxalutamide arm more frequently reported gastrointestinal adverse events, however, no patient discontinued treatment. In placebo group, 6 patients were lost during follow-up, and 2 patients died from acute respiratory distress syndrome. Here we demonstrate the hospitalization rate in proxalutamide treated men was reduced by 91% compared to usual care.

Final Results of a Randomized, Placebo-Controlled, Two-Arm, Parallel Clinical Trial of Proxalutamide for Hospitalized COVID-19 Patients: A Multiregional, Joint Analysis of the Proxa-Rescue AndroCoV Trial.

Background The role of androgens on COVID-19 is well established. Proxalutamide is a second-generation, non-steroidal antiandrogen (NSAA) with the highest antiandrogen potency among NSAAs and concurrent regulation of angiotensin-converting enzyme 2 (ACE2) expression and inflammatory response. Proxalutamide has been demonstrated to be effective to prevent hospitalizations in early COVID-19 in randomized clinical trials (RCTs). Conversely, in hospitalized COVID-19 patients, preliminary results from two different arms of an RCT (The Proxa-Rescue AndroCoV Trial) also demonstrated a reduction in all-cause mortality. This study aims to report the final, joint results of the two arms (North arm and South arm) of the Proxa-Rescue AndroCoV trial of the two arms (North and South arms) combined, and to evaluate whether COVID-19 response to proxalutamide was consistent across different regions (Northern Brazil and Southern Brazil). Materials and methods Upon randomization, hospitalized COVID-19 patients received either proxalutamide 300mg/day or placebo for 14 days, in addition to usual care, in a proxalutamide:placebo ratio of 1:1 in the North arm and 4:1 in the South arm (ratio was modified due to preliminary report of high drug efficacy). Datasets of the South and North arms were combined, and statistical analysis was performed for the overall study population. Proxalutamide was compared to placebo group for 14-day and 28-day recovery (discharge alive from the hospital) and mortality rates, and overall and post-randomization hospitalization stay. Results of proxalutamide and placebo groups were also compared between the North and South arms. Analysis was also performed stratified by sex and baseline WHO COVID Ordinary Score. Results A total of 778 subjects were included (645 from the North, 317 from the proxalutamide group and 328 from the placebo group; 133 from the South arm, 106 from the proxalutamide group and 27 from the placebo group). Recovery rate was 121% higher in proxalutamide than placebo group at day 14 [81.1% vs 36.6%; Recovery ratio (RecR) 2.21; 95% confidence interval (95% CI), 1.92-2.56; p<0.0001], and 81% higher at day 28 (98.1% vs 47.6%; RecR, 1.81; 95% CI, 1.61-2.03; p<0.0001). All-cause mortality rate was 80% lower in proxalutamide than placebo group at Day 14 [8.0% vs 39.2%; Risk ratio (RR), 0.20; 95% CI, 0.14-0.29; p<0.0001], and 78% lower at Day 28 (10.6% vs 48.2%; RR, 0.22; 95% CI 0.16-0.30). Post-randomization time-to-discharge was shorter in proxalutamide [median, 5 days; interquartile range (IQR), 3-8] than placebo group (median, 9 days; IQR, 6-14) (p<0.0001). Results were statistically similar between North and South arms for all measured outcomes. Males and females presented similar results in all outcomes. Patients that did not require oxygen use (scores 3 and 4) did not present statistically significant improvement in recovery and mortality rates, whereas scores 5 and 6 presented significant improvements in all outcomes (p<0.0001 for all). Conclusion Proxalutamide increased recovery rate, reduced mortality rate and shortened hospital stay in hospitalized COVID-19 patients. Results were similar between the two different arms, providing further consistency for the efficacy of proxalutamide when used in late-stage COVID-19.

EGAD! There is an ERAD doppelganger in the Golgi.

Disposal of membrane proteins in the late secretory pathway is thought to be exclusively facilitated by ESCRT-dependent lysosomal degradation. In this issue of The EMBO Journal, Schmidt et al define a previously uncharacterized endosome and Golgi-associated degradation (EGAD) pathway. This pathway, which has remarkable similarities to ERAD in the endoplasmic reticulum, operates in post-ER organelles via the proteasome and contributes to lipid homeostasis in eukaryotic cells.

Aneurysmal Bone Cyst of the Skull Base ­ Case Report

Introduction Aneurysmal bone cysts (ABCs) are pseudotumoral bone lesions of unknown etiology that are also hypervascularized, benign, and locally destructive. They are rare in the base of the skull. The present case report describes a case of aneurysmal bone cyst in the sella turcica. Case Report The present study was developed at the department of neurosurgery of the Hospital Universitário Professor Alberto Antunes of the Universidade Federal de Alagoas (HUPAA-AL, in the Portuguese acronym), Maceió, state of Alagoas, Brazil, and is accompanied by a review of the literature from the PubMed database. A 17-year-old female patient with bitemporal hemianopia and intense left hemicranial headache associated with symptoms from the cranial nerves contained in the cavernous sinus. Neuroimaging evidenced a large lesion in the suprasellar region with calcification foci, sellar erosion, and extension to the cavernous sinus. The patient was submitted to a partial lesion resection and the histopathological analysis showed an aneurysmal bone cyst. Conclusion A rare case of intracranial aneurysmal bone cyst, with the important differential diagnosis from pituitary adenoma.

Complicação tardia em biópsia hepática percutânea: é possível? / Late complication after percutaneous liver biopsy: is it possible?

A biópsia hepática percutânea, guiada ou não por ultrassonografia, consiste na retirada de fragmento de tecido hepático. É um procedimento invasivo para estudo histopatológico do órgão, propor terapêutica, avaliar resultados de tratamento e fazer acompanhamento após transplante em várias doenças hepáticas. A taxa de complicações deste procedimento está em torno de 1%, podendo citar dor, hipotensão, hemorragia, pneumotórax, peritonite biliar, bacteremia transitória e morte, e, na sua grande maioria, ocorre dentro das primeiras 48 horas. Relatamos o caso de um paciente do sexo masculino, 62 anos, com história de infecção crônica pelo vírus da hepatite C, submetido à biópsia hepática para estadiamento METAVIR. Após duas semanas, retornou ao serviço de urgência com quadro de abdome agudo hemorrágico, evoluindo a óbito. Com base no relato, os autores discutem as complicações da biópsia hepática, as causas de sangramento abdominal considerando a apresentação tardia do sangramento e a provável causa do óbito do paciente.

Liver biopsy, guided by ultrasonography or not, is an invasive procedure in which a hepatic tissue is removed for histopathological study of the organ, treatment planning, evaluate treatments results and follow-up after liver transplant in several hepatic diseases. Complication rate of the procedure is around 1%, including pain, hypotension, bleeding or hemorrhage, pneumothorax, biliary peritonitis transitory bacteremia and death, and most of them occur within 48 hours after the biopsy. We report a case about a male patient, 62 years old, with previous chronic hepatitis C virus infection, which underwent a liver biopsy for METAVIR staging. After two weeks, patient returns to the hospitals emergency service with hemorrhagic acute abdomen, leading to death. Based on the case report, authors discuss the complications of the liver biopsy, causes of abdominal bleeding considering its late manifestation and the patients probable cause of death.

The role of fibrinogen glycation in ATTR: evidence for chaperone activity loss in disease.

Transthyretin amyloidosis (ATTR) belongs to a class of disorders caused by protein misfolding and aggregation. ATTR is a disabling disorder of autosomal dominant trait, where transthyretin (TTR) forms amyloid deposits in different organs, causing dysfunction of the peripheral nervous system. We previously discovered that amyloid fibrils from ATTR patients are glycated by methylglyoxal. Even though no consensus has been reached about the actual role of methylglyoxal-derived advanced glycation end-products in amyloid diseases, evidence collected so far points to a role for protein glycation in conformational abnormalities, being ubiquitously found in amyloid deposits in Alzheimer's disease, dialysis-related amyloidosis and Parkinson's diseases. Human fibrinogen, an extracellular chaperone, was reported to specifically interact with a wide spectrum of stressed proteins and suppress their aggregation, being an interacting protein with TTR. Fibrinogen is differentially glycated in ATTR, leading to its chaperone activity loss. Here we show the existence of a proteostasis imbalance in ATTR linked to fibrinogen glycation by methylglyoxal.

Ácidos orgânicos na qualidade fisiológica de sementes de arroz / Effect of organic acids in physiological quality of rice

O alagamento do solo para o cultivo do arroz irrigado promove condições anaeróbias que favorecem a produção de ácidos orgânicos de cadeia curta, os quais podem ser tóxicos para a cultura. No presente trabalho, o objetivo foi avaliar a qualidade fisiológica de sementes de arroz irrigado cultivar 'IRGA 424', submetidas ao estresse por diferentes ácidos orgânicos, sendo selecionados aqueles formados no solo: acético, propiônico e butírico, utilizando-se as doses de 0, 4, 8, 12 e 16mM, nas quais as sementes foram embebidas por um período de 90 minutos e em seguida submetidas a testes de germinação e vigor (primeira contagem da germinação, comprimento e massa seca da parte aérea e raízes). Na avaliação da qualidade das sementes frente ao estresse por ácidos orgânicos, o comprimento das raízes foi mais eficiente para diferenciar a toxicidade nas doses de ácido acético, butírico e propiônico. O ácido butírico foi o mais prejudicial para sementes de arroz, afetando o desenvolvimento inicial das plântulas da cultivar 'IRGA 424'.

Flooding the soil for rice cultivation promotes anaerobic conditions that favor the production of short-chain organic acids, which can be toxic to the culture. This study aimed to evaluate the physiological quality of seeds of rice cultivar 'IRGA 424' subjected to stress by different organic acids. We studied three organic acids formed in the ground and five levels, namely: acetic, propionic, and butyric acid in doses 0, 4, 8, 12 and 16mM. The seeds were soaked in solutions and in the doses above, for a period of 90 minutes, after, were tested for germination and vigor (first count germination, shoot length and root and shoot dry mass and root). In evaluating the quality of the seed facing the stress by organic acids, the variable length of the roots was the most efficient to differentiate the toxicity by different doses of acetic acid, propionic and butyric. The butyric acid was the most damaging to rice seeds, affecting early development of seedlings of the cultivar 'IRGA 424'.

Potássio via recobrimento de sementes de soja: efeitos na qualidade fisiológica e no rendimento / Potassium via coating of soybean seeds: effects on physiological quality and yield

No presente trabalho, o objetivo foi avaliar a qualidade fisiológica e o rendimento de sementes de soja recobertas com diferentes fontes e concentrações de potássio. Utilizando a cultivar de soja 'CD 201', cujo delineamento experimental adotado foi inteiramente casualizado e os tratamentos constaram de um fatorial 2x5 (fontes de potássio: KCl e K2SO4 e níveis: 0, 5, 50, 500 e 5000mg L-1), totalizando dez tratamentos. As sementes foram recobertas com 2mL de solução de potássio + 2mL de Polímero (Sepiret®) + 2mL de Inoculante (Rizofix®) por quilo, totalizando 6mL kg-1 de semente. As variáveis analisadas foram primeira contagem da germinação, germinação, envelhecimento acelerado, teste de frio e comprimento da parte aérea e raiz; após a colheita, avaliou-se a qualidade fisiológica e o rendimento das sementes produzidas. O recobrimento de sementes de soja, cultivar 'CD 201', com cloreto e sulfato de potássio não influencia na germinação, mas o cloreto de potássio beneficia o desempenho das plântulas. O potássio via recobrimento de sementes com sulfato e cloreto de potássio até 5000mg L-1 de semente não interfere no seu rendimento, porém o sulfato de potássio produz sementes de melhor qualidade.

This study aimed to evaluate the physiological quality and yield of soybean seeds coated with different sources and concentrations of potassium. Using soybean cultivar 'CD 201'. The experimental design was completely randomized and the treatments were a factorial 2x5 (sources of potassium: KCl and K2SO4 and levels: 0, 5, 50, 500 and 5000mg L-1), a total of ten treatments, with four replications. The seeds were covered with 2 ml of solution of potassium 2mL + Polymer (Sepiret®) + 2mL of inoculants (Rizofix®) per kg of seed, totaling 6mL kg-1 seeds. The variables analyzed after treatment of the seeds were first count of germination, germination, accelerated aging, cold test and length of shoot and root. After harvest, we evaluated the physiological seed quality and seed yield. It is concluded that the coating of soybean seeds, cultivar 'CD 201', chloride and potassium sulfate does not influence the germination of treated seed, however potassium positively influences the performance of seedlings. The seed coating by potassium sulphate and potassium chloride to 5000mg L-1 seed do not interfere with seed yield, but the potassium sulphate seed produces higher quality than chloride.

Experimental study of coexistence of multi-band OFDM-UWB and OFDM-baseband signals in long-reach PONs using directly modulated lasers.

Transmission of coexisting Orthogonal Frequency Division Multiplexing (OFDM)-baseband (BB) and multi-band OFDM-ultra-wideband (UWB) signals along long-reach passive optical networks using directly modulated lasers (DML) is experimentally demonstrated.When optimized modulation indexes are used, bit error ratios not exceeding 5 × 10⁻4 can be achieved by all (OFDM-BB and three OFDM-UWB sub-bands) signals for a reach of 100 km of standard single-mode fiber (SSMF) and optical signal-to-noise ratios not lower than 25dB@0.1 nm. It is experimentally shown that, for the SSMF reach of 100km, the optimized performance of coexisting OFDM-BB and OFDM-UWB signals is mainly imposed by the combination of two effects: the SSMF dispersion-induced nonlinear distortion of the OFDM-UWB signals caused by the OFDM-BB and OFDM-UWB signals, and the further degradation of the OFDM-UWB signals with higher frequency, due to the reduced DML bandwidth.

In-band crosstalk penalties in optical networks with narrow optical and electric filtering.

Optical cross-connects (OXC) introduce crosstalk and intersymbolic interference due to filtering in the optical signal. We show experimentally and by simulation that higher optical signal-to-noise ratio (OSNR) penalties are obtained for the combined effect of both impairments compared to the sum of the penalties taken independently. Furthermore, the Modified Chernoff Bound is applied for performance estimation, with good accuracy and fast computation time. By using the correct OSNR penalty, the maximum number of cascaded OXC is reduced by 17% relatively to when the penalties are considered separately, for a 2 dB maximum penalty.

High promoter methylation levels of APC predict poor prognosis in sextant biopsies from prostate cancer patients.

PURPOSE: Prostate cancer is a highly prevalent malignancy and constitutes a major cause of cancer-related morbidity and mortality. Owing to the limitations of current clinical, serologic, and pathologic parameters in predicting disease progression, we sought to investigate the prognostic value of promoter methylation of a small panel of genes by quantitative methylation-specific PCR (QMSP) in prostate biopsies. EXPERIMENTAL DESIGN: Promoter methylation levels of APC, CCND2, GSTP1, RARB2, and RASSF1A were determined by QMSP in a prospective series of 83 prostate cancer patients submitted to sextant biopsy. Clinicopathologic data [age, serum prostate-specific antigen (PSA), stage, and Gleason score] and time to progression and/or death from prostate cancer were correlated with methylation findings. Log-rank test and Cox regression model were used to identify which epigenetic markers were independent predictors of prognosis. RESULTS: At a median follow-up time of 45 months, 15 (18%) patients died from prostate cancer, and 37 (45%) patients had recurrent disease. In univariate analysis, stage and hypermethylation of APC were significantly associated with worse disease-specific survival, whereas stage, Gleason score, high diagnostic serum PSA levels, and hypermethylation of APC, GSTP1, and RASSF1A were significantly associated with poor disease-free survival. However, in the final multivariate analysis, only clinical stage and high methylation of APC were significantly and independently associated with unfavorable prognosis, i.e., decreased disease-free and disease-specific survival. CONCLUSIONS: High-level APC promoter methylation is an independent predictor of poor prognosis in prostate biopsy samples and might provide relevant prognostic information for patient management.

Mecanismos associados à tolerância ao alumínio em plantas / Associated mechanisms of aluminum tolerance in plants

A toxicidade do alumínio é um dos principais fatores limitantes do desenvolvimento das plantas emsolos ácidos. Pelo fato da utilização de corretivos da acidez do solo não ser a estratégia mais viável emmuitas situações com solos ácidos (por razões técnicas e econômicas), o desenvolvimento de genótipostolerantes ao Al tem sido o caminho mais focado, assim a investigação dos mecanismos de tolerânciabem como as bases genéticas da tolerância ao Al têm merecido atenção especial pela pesquisa científica.Nos últimos anos, foi gerado um significativo progresso no entendimento das bases dos mecanismos detolerância ao Al, assim como no desenvolvimento de cultivares mais adaptados as condições de solosácidos. Os mecanismos de tolerância ao Al conhecidos se resumem basicamente em duas classes: os queagem no sentido de expulsar o Al depois de absorvido ou de impedir sua entrada pela raiz e os mecanismosde desintoxicação, complexando o Al em organelas específicas da planta, principalmente nos vacúolos.Em inúmeras espécies, mecanismos fisiológicos tem sido reportados como responsáveis pela ativaçãode ácidos orgânicos (principalmente citrato e malato) que atuam como agentes quelantes do Al, porémmuitos processos ainda não são bem entendidos e esclarecidos. Atualmente, se começa a entendermelhor um segundo mecanismo de tolerância ao Al que envolve a desintoxicação interna do Al atravésda complexação por ácidos orgânicos e o seqüestro destes complexos pelos vacúolos. Outros mecanismospotenciais são alvo de especulações e discussões.

Aluminum toxicity is one of the major limiting factor regarding plant development in acid soils. The use of liming for correcting soil pH is not viable for some of acid soil areas (technique or economic reasons),making the development of Al tolerant genotypes the best alternative. Thus, the tolerance mechanisms as well as the genetic basis of Al tolerance has deserved special attention in the scientific community. In the last years, a significant progress has been achieved towards these goals, as well as in developing cultivars adapted to acid soils. The Al tolerance mechanisms are divided basically in two classes: the exclusion mechanisms that act after absorption or blocking its entry in the root system and those involved in detoxification, complexing the Al in specific organelles, mainly in the vacuoles. In man yspecies, physiological mechanisms have been reported as responsible for the activation of organic acids(mainly citrate and malate) that act as Al quelating agents, however many process are not yet understood and cleared. Currently, the basis for the internal detoxification is becoming clear through organic acid complexes and there sequestering by the vacuoles. Other potential mechanisms are the target for discussions

8q gain is an independent predictor of poor survival in diagnostic needle biopsies from prostate cancer suspects.

PURPOSE: The main procedure to confirm a suspected diagnosis of prostate cancer is histologic analysis of ultrasound-guided sextant prostate biopsies. As it is difficult to reliably assess tumor stage and grade in such minute samples, the clinical significance of some tumor foci remains unclear. Genetic markers that could augment pretreatment prognostic information would improve the clinical management of the disease. EXPERIMENTAL DESIGN: We have analyzed by comparative genomic hybridization a consecutive series of prostate needle biopsies obtained prospectively from 100 prostate cancer suspects. For 25 of these patients, a second independent biopsy core was analyzed to assess possible tumor heterogeneity. Additionally, a three-color fluorescent in situ hybridization assay was done in paraffin-embedded biopsy cores to validate the comparative genomic hybridization findings and to confirm their prognostic value. RESULTS: Sixty-one of 100 biopsy samples had morphologic evidence of prostate cancer and 41 (67%) of these displayed genomic copy number changes as opposed to none of the morphologically normal biopsies. The presence of losses, amplifications, and the total number of genomic imbalances were significantly associated with poorly differentiated tumors. Kaplan-Meier curves with log-rank test showed that patients whose tumors displayed 8q gains had a significantly worse survival even when tumor grade was taken into account (P = 0.008). Restricting the analysis to cases with Gleason score 7, the most troublesome category in terms of prognostic information, gains at 8q were still significantly associated with poor survival (P = 0.011), something that was confirmed by fluorescent in situ hybridization in an independent series of biopsies with much longer follow-up time (P = 0.023). CONCLUSIONS: We show that whole genomic information can be obtained from minute needle biopsies of prostate cancer suspects and that genetic data can provide additional prognostic information before a therapeutic decision is taken.

Quantitative RARbeta2 hypermethylation: a promising prostate cancer marker.

Retinoic acid receptor beta2 (RARbeta2) is a tumor suppressor gene frequently hypermethylated in several human neoplasms. To further characterize this epigenetic alteration in prostate cancer progression, we examined tumor tissue from 118 patients with prostate carcinoma (PCa), 38 paired high-grade prostatic intraepithelial neoplasias (HGPIN), and non-neoplastic prostate tissue from 30 patients with benign prostate hyperplasia (BPH), using quantitative methylation-specific PCR. We found RARbeta2 hypermethylation in 97.5% of PCa, 94.7% of HGPIN, and 23.3% of BPH. Methylation levels were significantly higher in PCa compared with HGPIN and BPH (P < 0.00001). By establishing an empiric cutoff value, we were able to discriminate between neoplastic and non-neoplastic tissue, with 94.9% sensitivity and 100% specificity. Moreover, RARbeta2 methylation levels correlated with higher pathological stage (r = 0.30, P = 0.0009). This quantitative assay represents a novel and promising molecular marker that may augment current approaches for prostate cancer detection.