Enhancing Prostate Tumor Biobanking Reliability with Improved Sampling Technique and Histological Characterization.

Acquiring fresh and well-characterized tumor tissue samples is critical for conducting high-quality "omics" studies. However, it can be particularly challenging in the context of prostate cancer (PC) due to the unique nature of this organ and the high heterogeneity associated with this tumor. On the other hand, histopathologically characterizing samples before their storage without causing significant tissue alterations is also an intriguing challenge. In this context, we present a new method for acquiring, mapping, characterizing, and micro-dissecting resected prostate tissue based on anatomopathological criteria. Unlike previously published protocols, this method reduces the time required for histopathological analysis of the prostate specimen without compromising its structure, which is crucial for assessing surgical margins. Furthermore, it enables the delineation and micro-macro dissection of fresh prostate tissue samples, with a focus on histological tumor areas defined by pathological criteria such as Gleason score, precursor lesions (high-grade prostatic intraepithelial neoplasia - PIN), and inflammatory lesions (prostatitis). These samples are then stored in a Biobank for subsequent research analyses.

Bladder cancer patients have increased NETosis and impaired DNaseI-mediated NET degradation that can be therapeutically restored in vitro.

Background: Neutrophils, key players of the immune system, also promote tumor development through the formation of neutrophil extracellular traps (NETs) in a process called NETosis. NETs are extracellular networks of DNA, histones and cytoplasmic and granular proteins (calprotectin, myeloperoxidase, elastase, etc.) released by neutrophils upon activation. NETs regulate tumor growth while promoting angiogenesis and invasiveness, and tumor cells also stimulate NETosis. Although NETosis seems to be increased in cancer patients, an increase of NETs in plasma may also be mediated by an impaired degradation by plasma DNaseI, as evidenced in several immunological disorders like lupus nephritis. However, this has never been evidenced in bladder cancer (BC) patients. Herein, we aimed to evaluate the occurrence of increased NETosis in plasma and tumor tissue of BC patients, to ascertain whether it is mediated by a reduced DNaseI activity and degradation, and to in vitro explore novel therapeutic interventions. Methods: We recruited 71 BC patients from whom we obtained a plasma sample before surgery and a formalin-fixed paraffin embedded tumor tissue sample, and 64 age- and sex-matched healthy controls from whom we obtained a plasma sample. We measured NETs markers (cell-free fDNA, calprotectin, nucleosomes and neutrophil elastase) and the DNaseI activity in plasma with specific assays. We also measured NETs markers in BC tissue by immunofluorescence. Finally, we evaluated the ability of BC and control plasma to degrade in vitro-generated NETs, and evaluated the performance of the approved recombinant human DNaseI (rhDNaseI, Dornase alfa, Pulmozyme®, Roche) to restore the NET-degradation ability of plasma. In vitro experiments were performed in triplicate. Statistical analysis was conducted with Graphpad (v.8.0.1). Results: NETosis occurs in BC tissue, more profusely in the muscle-invasive subtype (P<0.01), that with the worst prognosis. Compared to controls, BC patients had increased NETosis and a reduced DNaseI activity in plasma (P<0.0001), which leads to an impairment to degrade NETs (P<0.0001). Remarkably, this can be therapeutically restored with rhDNaseI to the level of healthy controls. Conclusion: To the best of our knowledge, this is the first report demonstrating that BC patients have an increased NETosis systemically and in the tumor microenvironment, in part caused by an impaired DNaseI-mediated NET degradation. Remarkably, this defect can be therapeutically restored in vitro with the approved Dornase alfa, thus Pulmozyme® could become a potential therapeutic tool to locally reduce BC progression.

Multiple plasma membrane reporters discern LHFPL5 region that blocks trafficking to the plasma membrane.

The mechano-electrical transduction (MET) channel of the inner ear receptor cells, termed hair cells, is a protein complex that enables our senses of hearing and balance. Hair cell MET requires an elaborate interplay of multiple proteins that form the MET channel. One of the MET complex components is the transmembrane protein LHFPL5, which is required for hair cell MET and hearing. LHFPL5 is thought to form a multi-protein complex with other MET channel proteins, such as PCDH15, TMIE, and TMC1. Despite localizing to the plasma membrane of stereocilia, the mechanosensing organelles of hair cells, LHFPL5 requires its binding partner within the MET complex, PCDH15, to localize to the stereocilia tips in hair cells and to the plasma membrane in heterologous cells. Using the Aquaporin 3-tGFP reporter (AGR) for plasma membrane localization, we found that a region within extracellular loop 1, which interacts with PCDH15, precludes the trafficking of AGR reporter to the plasma membrane in heterologous cell lines. Our results suggest that the presence of protein partners may mask endoplasmic reticulum retention regions or enable the proper folding and trafficking of the MET complex components, to facilitate expression of the MET complex at the stereocilia membrane.

Targeting TGF-ß signaling in the multiple myeloma microenvironment: Steering CARs and T cells in the right direction.

Multiple myeloma (MM) remains a lethal hematologic cancer characterized by the expansion of transformed plasma cells within the permissive bone marrow (BM) milieu. The emergence of relapsed and/or refractory MM (RRMM) is provoked through clonal evolution of malignant plasma cells that harbor genomic, metabolic and proteomic perturbations. For most patients, relapsed disease remains a major cause of overall mortality. Transforming growth factors (TGFs) have pleiotropic effects that regulate myelomagenesis as well as the emergence of drug resistance. Moreover, TGF-ß modulates numerous cell types present with the tumor microenvironment, including many immune cell types. While numerous agents have been FDA-approved over the past 2 decades and significantly expanded the treatment options available for MM patients, the molecular mechanisms responsible for drug resistance remain elusive. Multiple myeloma is uniformly preceded by a premalignant state, monoclonal gammopathy of unknown significance, and both conditions are associated with progressive deregulation in host immunity characterized by reduced T cell, natural killer (NK) cell and antigen-presenting dendritic cell (DC) activity. TGF-ß promotes myelomagenesis as well as intrinsic drug resistance by repressing anti-myeloma immunity to promote tolerance, drug resistance and disease progression. Hence, repression of TGF-ß signaling is a prerequisite to enhance the efficacy of current and future immunotherapeutics. Novel strategies that incorporate T cells that have been modified to express chimeric antigen receptor (CARs), T cell receptors (TCRs) and bispecific T cell engagers (BiTEs) offer promise to block TGF-ß signaling, overcome chemoresistance and enhance anti-myeloma immunity. Here, we describe the effects of TGF-ß signaling on immune cell effectors in the bone marrow and emerging strategies to overcome TGF-ß-mediated myeloma growth, drug resistance and survival.

Assessing the Underestimation of HIV Risk Infection among Young Men Who Have Sex with Men in Argentina.

The aim of this study is to describe the discordance between the self-perceived risk and actual risk of HIV among young men who have sex with men (YMSM) and its associated factors. An online, cross-sectional study was conducted with 405 men recruited from an Argentinian NGO in 2017. Risk discordance (RD) was defined as the expression of the underestimation of risk, that is, as a lower self-perception of HIV risk, as measured with the Perceived Risk of HIV Scale, than the current risk of HIV infection, as measured by the HIV Incidence Risk Index. Multivariate logistic regression models were used to analyze the associations between the RD and the explanatory variables. High HIV risk was detected in 251 (62%), while 106 (26.2%) showed high self-perceived risk. RD was found in 230 (56.8%) YMSM. The predictors that increased RD were consistent condom use with casual partners (aOR = 3.8 [CI 95:1.5-11.0]), the use of Growler to meet partners (aOR = 10.38 [CI 95:161-121.94]), frequenting gay bars (aOR = 1.9 [95% CI:1.1-3.5]) and using LSD (aOR = 5.44 [CI 95:1.32-30.29]). Underestimation of HIV risk in YMSM is associated with standard HIV risk behavior and modulated by psychosocial aspects. Thus, prevention campaigns aimed at YMSM should include these factors, even though clinical practice does not. Health professionals should reconsider adapting their instruments to measure the risk of HIV in YMSM. It is unknown what score should be used for targeting high-risk YMSM, so more research is needed to fill this gap. Further research is needed to assess what score should be used for targeting high-risk in YMSM.

[Predictions of three mathematical models related with the COVID-19 Vaccination Strategy in Spain. June 2021.] / Predicciones de tres modelos matemáticos en relación a la estrategia de vacunación frente a la COVID-19 en España. Junio de 2021.

The Ministry of Health has coordinated three studies that have estimated the impact of the COVID-19 Vaccination Strategy in Spain. The models aim to help how to establish priority population groups for vaccination, in an initial context of dose limitation. With the same epidemiological and vaccine information, the results of this three different mathematical models point in the same direction: combined with physical distancing, staggered vaccination, starting with the high risk groups, would prevent 60% of infections, 42% of hospitalizations and 60% of mortality in the population. These models, which can be adapted to the new available scientific evidence, are dynamic and powerful tools for the evaluation and adjustment of immunization programs, promoting research on this field, and helping to achieve more efficient results in health.

El Ministerio de Sanidad ha coordinado tres estudios que han estimado el impacto de la Estrategia de Vacunación frente a COVID-19 en España. El objetivo era que los modelos ayudaran a establecer los grupos de población prioritarios para la vacunación, en un contexto inicial de limitación de dosis. A partir de la misma información epidemiológica y de vacunas se han elaborado tres modelos matemáticos distintos cuyos resultados apuntan en la misma dirección: combinada con el distanciamiento físico, la vacunación escalonada, empezando por los grupos de mayor riesgo de complicaciones, evitaría el 60% de las infecciones, el 42% de las hospitalizaciones y el 60% de la mortalidad en la población. Estos modelos, que pueden adaptarse a la nueva evidencia científica disponible, son herramientas dinámicas y potentes para la evaluación y el ajuste de los programas de vacunación, impulsando el desarrollo de este campo de investigación, y ayudando a lograr resultados más eficientes en salud.

Predicciones de tres modelos matemáticos en relación a la estrategia de vacunación frente a la COVID-19 en España. Junio de 2021 / Predictions of three mathematical models related with the COVID-19 Vaccination Strategy in Spain. June 2021

El Ministerio de Sanidad ha coordinado tres estudios que han estimado el impacto de la Estrategia de Vacunación frente a COVID-19 en España. El objetivo era que los modelos ayudaran a establecer los grupos de población prioritarios para la vacunación, en un contexto inicial de limitación de dosis. A partir de la misma información epidemiológica y de vacunas se han elaborado tres modelos matemáticos distintos cuyos resultados apuntan en la misma dirección: combinada con el distanciamiento físico, la vacunación escalonada, empezando por los grupos de mayor riesgo de complicaciones, evitaría el 60% de las infecciones, el 42% de las hospitalizaciones y el60% de la mortalidad en la población. Estos modelos, que pueden adaptarse a la nueva evidencia científica disponible, son herramientasdinámicas y potentes para la evaluación y el ajuste de los programas de vacunación, impulsando el desarrollo de este campo de investigación, y ayudando a lograr resultados más eficientes en salud.(AU)

The Ministry of Health has coordinated three studies that have estimated the impact of the COVID-19 Vaccination Strategy in Spain. The models aim to help how to establish priority population groups for vaccination, in aninitial context of dose limitation. With the same epidemiological and vaccine information, the results of this three different mathematical models point in the same direction: combined with physical distancing, staggered vaccination, starting with the high risk groups, would prevent 60% of infections, 42% of hospitalizations and 60% of mortality in the population. These models, which can be adapted to the new available scientific evidence, are dynamic and powerful tools for the evaluation and adjustment of immunization programs, promoting research on this field, and helping to achieve more efficient results in health.(AU)

Cold loop polypectomy perforation of a tiny colon polyp.

We report the case of a 61-year-old woman having corticoid treatment with corticosteroids for polyarthralgia, who underwent a post-polypectomy surveillance colonoscopy, identifying a 5-mm diameter, flat-elevated polyp in the proximal transverse colon (Paris 0-IIa).

Using chimeric antigen receptor T-cell therapy to fight glioblastoma multiforme: past, present and future developments.

INTRODUCTION: Glioblastoma multiforme (GBM) constitutes one of the deadliest tumors to afflict humans, although it is still considered an orphan disease. Despite testing multiple new and innovative therapies in ongoing clinical trials, the median survival for this type of malignancy is less than two years after initial diagnosis, regardless of therapy. One class of promising new therapies are chimeric antigen receptor T cells or CAR-T which have been shown to be very effective at treating refractory liquid tumors such as B-cell malignancies. However, CAR-T effectivity against solid tumors such as GBM has been limited thus far. METHODS: A Pubmed, Google Scholar, Directory of Open Access Journals, and Web of Science literature search using the terms chimeric antigen receptor or CAR-T, GBM, solid tumor immunotherapy, immunotherapy, and CAR-T combination was performed for publication dates between January 1987 and November 2021. RESULTS: In the current review, we present a comprehensive list of CAR-T cells developed to treat GBM, we describe new possible T-cell engineering strategies against GBM while presenting a short introductory history to the reader regarding the origin(s) of this cutting-edge therapy. We have also compiled a unique list of anti-GBM CAR-Ts with their specific protein sequences and their functions as well as an inventory of clinical trials involving CAR-T and GBM. CONCLUSIONS: The aim of this review is to introduce the reader to the field of T-cell engineering using CAR-Ts to treat GBM and describe the obstacles that may need to be addressed in order to significantly delay the relentless growth of GBM.

Healthy myeloid-derived suppressor cells express the surface ectoenzyme Vanin-2 (VNN2).

Study of human monocytic Myeloid-Derived Suppressor cells Mo-MDSC (CD14+ HLA-DRneg/low) has been hampered by the lack of positive cell-surface markers. In order to identify positive markers for Mo-MDSC, we performed microarray analysis comparing Mo-MDSC cells from healthy subjects versus CD14+ HLA-DRhigh monocytes. We have identified the surface ectoenzyme Vanin-2(VNN2) protein as a novel biomarker highly-enriched in healthy subjects Mo-MDSC. Indeed, healthy subjects Mo-MDSC cells expressed 68 % VNN2, whereas only 9% VNN2 expression was observed on CD14+ HLA-DRhigh cells (n = 4 p < 0.01). The top 10 percent positive VNN2 monocytes expressed CD33 and CD11b while being negative for HLA-DR, CD3, CD15, CD19 and CD56, consistent with a Mo-MDSC phenotype. CD14+VNN2high monocytes were able to inhibit CD8 T cell proliferation comparably to traditional Mo-MDSC at 51 % and 48 % respectively. However, VNN2 expression on CD14+ monocytes from glioma patients was inversely correlated to their grade. CD14+VNN2high monocytes thus appear to mark a monocytic population similar to Mo-MDSC only in healthy subjects, which may be useful for tumor diagnoses.

Breakage of CRISPR/Cas9-Induced Chromosome Bridges in Mitotic Cells.

Chromosomal instability, the most frequent form of plasticity in cancer cells, often proceeds through the formation of chromosome bridges. Despite the importance of these bridges in tumor initiation and progression, debate remains over how and when they are resolved. In this study, we investigated the behavior and properties of chromosome bridges to gain insight into the potential mechanisms underlying bridge-induced genome instability. We report that bridges may break during mitosis or may remain unbroken until the next interphase. During mitosis, we frequently observed discontinuities in the bridging chromatin, and our results strongly suggest that a substantial fraction of chromosome bridges are broken during this stage of the cell cycle. This notion is supported by the observation that the chromatin flanking mitotic bridge discontinuities is often decorated with the phosphorylated form of the histone H2AX, a marker of DNA breaks, and by MDC1, an early mediator of the cell response to DNA breaks. Also, free 3'OH DNA ends were detected in more than half of the bridges during the final stages of cell division. However, even if detected, the DNA ends of broken bridges are not repaired in mitosis. To investigate whether mitotic bridge breakage depends on mechanical stress, we used experimental models in which chromosome bridges with defined geometry are formed. Although there was no association between spindle pole separation or the distance among non-bridge kinetochores and bridge breakage, we found a direct correlation between the distance between bridge kinetochores and bridge breakage. Altogether, we conclude that the discontinuities observed in bridges during mitosis frequently reflect a real breakage of the chromatin and that the mechanisms responsible for chromosome bridge breakage during mitosis may depend on the separation between the bridge kinetochores. Considering that previous studies identified mechanical stress or biochemical digestion as possible causes of bridge breakage in interphase cells, a multifactorial model emerges for the breakage of chromosome bridges that, according to our results, can occur at different stages of the cell cycle and can obey different mechanisms.

[The third version of COPSOQ-Istas21. An updated international instrument for the workplace.] / La tercera versión de COPSOQ-Istas21. Un instrumento internacional actualizado para la prevención de riesgos psicosociales en el trabajo.

OBJECTIVE: The Copenhagen Psychosocial Questio-nnaire (COPSOQ) is one of the most widely used in research and psychosocial risk assessment in the workplace. The adaptation of the third international COPSOQ version to Spain is described and the evidence of its validity and reliability presented. METHODS: Most of the items were already part of the previous versions I and II. The translation of the new items was done by means of translation/reverse translation. The questionnaire was included in the Psychosocial Risk Survey 2016, a cross-sectional study of a representative sample of the wage-earning population in Spain (N=1,807). Descriptive statistics, internal consistency, floor and ceiling effects and factor structure were analysed. Prevalence Ratios adjusted by age, sex and occupational class (aPR) to mental health, general health and job satisfaction were calculated. Finally, population reference values were calculated for all dimensions of the instrument. RESULTS: The questionnaire showed an excellent factorial structure. All scales, except one, showed α of Cronbach >0.70. Comparing the COPSOQ-Istas21 III scales with their international references, Cronbach's α were higher and the ceiling and floor effects were lower; i.e. Organisational Justice: α=0.85 vs 0.74, ceiling and floor 1.6 and 9.4 vs 3.3 and 12.8. The aPR between all the psychosocial dimensions and the Mental Health, General Health and Job Satisfaction were in the expected direction and showed an association gradient. CONCLUSIONS: COPSOQ-Istas21 version III presents psychometric properties analogous or better than the original in English language, and good indicators of validity and reliability, to be used in research and psychosocial risk assessment at the workplace in Spain.

OBJETIVO: El Copenhagen Psychosocial Questionnaire (COPSOQ) es uno de los más utilizados en investigación y evaluación de riesgos psicosociales en el trabajo. En este artículo se describió la adaptación de la tercera versión internacional a España y se presentaron las pruebas de su validez y fiabilidad. METODOS: La mayoría de los ítems ya formaban parte de las versiones anteriores I y II. La traducción de los nuevos ítems fue realizada mediante traducción/traducción inversa. El cuestionario se incluyó en la Encuesta de Riesgos Psicosociales de 2016, estudio transversal de una muestra representativa de la población asalariada en España (N=1.807). Se analizaron los estadísticos descriptivos, consistencia interna, efectos suelo y techo y estructura factorial. Se calcularon las Razones de Prevalencia ajustadas por edad, sexo y clase ocupacional (aRP) con salud mental, salud general y satisfacción laboral. Finalmente, se calcularon los valores de referencia poblacionales para todas las dimensiones del instrumento. RESULTADOS: El cuestionario mostró una excelente estructura factorial. Todas las escalas, excepto una, mostraron α de Cronbach >0,70. Comparando las escalas de COPSOQ-Istas21 III con sus referentes internacionales, las α de Cronbach fueron más altas y los efectos techo y suelo menores (por ejemplo, Justicia organizacional: α=0,85 vs 0,74, techo y suelo 1,6 y 9,4 vs 3,3 y 12,8). Las aRP entre todas las dimensiones psicosociales y las dimensiones de Salud mental, Salud general y Satisfacción, fueron en la dirección esperada y mostraron un gradiente de asociación. CONCLUSIONES: COPSOQ-Istas21 versión III presenta propiedades psicométricas análogas o mejores al original en lengua inglesa y buenos indicadores de validez y fiabilidad para ser usado en investigación y evaluación de riesgos psicosociales laborales en España.

Evaluation of the optical criteria for sessile serrated lesions of the colon: A prospective study on a colorectal cancer screening population.

Background and study aims We aimed to describe the presence and combination of Hazewinkel's optical diagnosis (OD) criteria for sessile serrated lesions (SSL), determining which lesion characteristics increase the probability of a correct OD, with a focus on diminutive lesions. Patients and methods This was a prospective study describing the presence of Hazewinkel's OD criteria for SSL in lesions found in consecutive CRC screening colonoscopies. The presence of each OD criterion and their diagnostic combinations in SSL, related to the lesion's NBI International Colorectal Endoscopic (NICE) classification category, size, and location, were described. The presence of two or more optical criteria was considered diagnostic of SSL. The OD was compared to pathology as the gold standard. Results Seventy-nine SSLs (5.6 %) were diagnosed. Cloud-like appearance was the most prevalent OD criterion (35, 44.3 %). OD criteria were more frequently identified in NICE type 1, ≥ 10 mm, and proximal lesions. Only 26 SLLs fulfilled the OD criteria (sensitivity 32.9 %, 95 % CI 29.1 %-36.7 %). The sensitivity for diminutive SSL was 14.7 %, (95 % CI 11.9 %-17.6 %). Eighty-five lesions were optically diagnosed as SSL. However, only in 26 SSL was this the definitive diagnosis (positive predictive value 30.6 %, 95 % CI 26.9 %-34.3 %). Size > 5 mm and proximal location increased the probability of a correct diagnosis. The overall accuracy of the optical criteria was 92.0 % (95 % CI, 89.8 %-94.2 %). Conclusions The Hazewinkel's optical criteria are not reliable for a positive diagnosis of SSL, particularly for diminutive lesions.

A Liquid Biopsy to Assess Brain Tumor Recurrence: Presence of Circulating Mo-MDSC and CD14+ VNN2+ Myeloid Cells as Biomarkers That Distinguish Brain Metastasis From Radiation Necrosis Following Stereotactic Radiosurgery.

BACKGROUND: Brain metastases (BM) are the most common type of brain tumor malignancy in the US. They are also the most common indication for stereotactic radiosurgery (SRS). However, the incidence of both local recurrence and radiation necrosis (RN) is increasing as treatments improve. MRI imagery often fails to differentiate BM from RN; thus, patients must often undergo surgical biopsy or resection to obtain a definitive diagnosis. OBJECTIVE: To hypothesize that a marker of immunosuppression might serve as a surrogate marker to differentiate patients with active vs inactive cancer-including RN. METHODS: We thus purified and quantified Monocytic Myeloid-Derived Suppressor Cells (Mo-MDSC) by flow cytometry in patients proven by biopsy to represent BM or RN. RESULTS: We report the utility of the previously reported HLA-Dr-Vnn2 Index or DVI to discriminate recurrent BM from RN using peripheral blood. The presence of CD14+ HLA-DRneg/low Mo-MDSC is significantly increased in the peripheral blood of patients with brain metastasis recurrence compared to RN (Average 61.5% vs 7%, n = 10 and n = 12, respectively, P < .0001). In contrast, expression of VNN2 on circulating CD14+ monocytes is decreased in BM patients compared to patients with RN (5.5% vs 26.5%, n = 10 and n = 12, respectively, P = .0008). In patients with biopsy confirmed recurrence of brain metastasis, the average DVI was 11.65, whereas the average DVI for RN patients was consistently <1 (Avg. of 0.17). CONCLUSION: These results suggest that DVI could be a useful diagnostic tool to differentiate recurrent BM from RN using a minimally invasive blood sample.

Design and Refinement of a Data Quality Assessment Workflow for a Large Pediatric Research Network.

BACKGROUND: Clinical data research networks (CDRNs) aggregate electronic health record data from multiple hospitals to enable large-scale research. A critical operation toward building a CDRN is conducting continual evaluations to optimize data quality. The key challenges include determining the assessment coverage on big datasets, handling data variability over time, and facilitating communication with data teams. This study presents the evolution of a systematic workflow for data quality assessment in CDRNs. IMPLEMENTATION: Using a specific CDRN as use case, the workflow was iteratively developed and packaged into a toolkit. The resultant toolkit comprises 685 data quality checks to identify any data quality issues, procedures to reconciliate with a history of known issues, and a contemporary GitHub-based reporting mechanism for organized tracking. RESULTS: During the first two years of network development, the toolkit assisted in discovering over 800 data characteristics and resolving over 1400 programming errors. Longitudinal analysis indicated that the variability in time to resolution (15day mean, 24day IQR) is due to the underlying cause of the issue, perceived importance of the domain, and the complexity of assessment. CONCLUSIONS: In the absence of a formalized data quality framework, CDRNs continue to face challenges in data management and query fulfillment. The proposed data quality toolkit was empirically validated on a particular network, and is publicly available for other networks. While the toolkit is user-friendly and effective, the usage statistics indicated that the data quality process is very time-intensive and sufficient resources should be dedicated for investigating problems and optimizing data for research.

Tumor Budding: Prognostic Value in Muscle-invasive Bladder Cancer.

OBJETIVES: To assess if "tumor budding" (TB) behaves as a poor prognostic factor in muscle-invasive bladder carcinoma (MIBC). TB is the presence of tumor cells isolated or in small groups of fewer than 5 cells located at the tumor invasion front. MATERIAL AND METHODS: Retrospective study of 106 patients with MIBC who underwent radical cystectomy. A cytokeratin AE1/AE3 immunostaining was applied to identify and quantify TB by the "hot-spot" method. The variables evaluated were: age, gender, Tumour, Node, Metastasis Classification (TNM) stage, associated Carcinoma in situ, differentiation degree, tumor size, tumor location, lymphatic, venous or perineural invasion, p53, Ki67, molecular subtype (basal/luminal) and chemotherapy. Main variables were overall and cancer-specific survival. RESULTS: The mean follow-up time was 47 ± 46.45 months. The mean TB count was 32.3 ± 25.9 "buds." The ROC curve established 14 "buds" as the cut-off point: the median survival rate for the "low-grade TB" group (≤14 "buds") was 69.5 months, and for the "high-grade TB" group (>14 "buds") was 18.5 months (P= .003). In the multivariate analysis, independent predictive variables regarding mortality were: age, TB, and TNM stage. Patients with more than 14 "buds" had 2.27 times more risk of mortality, 95%CI:1.19-4.34, P = .013. In addition, the risk of mortality rises progressively as the number of "buds" increases, at a rate of 2% per "bud." CONCLUSION: According to our results, TB becomes an independent predictor factor for cancer-specific mortality in MIBC, with a cut-off point of 14 "buds."

Carcinoma tubuloquístico. Una entidad infrecuente en la patología tumoral renal / Tubulocystic renal cell carcinoma. A rare entity in neoplastic renal pathology

El carcinoma renal tubuloquístico es una neoplasia infrecuente dentro de la patología tumoral renal. En el presente caso comentamos la evolución de un paciente con cólicos nefríticos de repetición en el que durante el seguimiento ecográfico se descubrió una lesión nodular de aspecto quístico. Posteriormente la lesión aumentó de tamaño y se decidió nefrectomía parcial (tumorectomía). En el examen histológico e inmunohistoquímico se estableció el diagnóstico de carcinoma renal tubuloquístico

Tubulocystic renal carcinoma is an uncommon neoplasm. We present a case of a patient presenting with multiple renal colic. A nodular cystic lesion was an incidental sonographic finding which increased in size during subsequent follow-ups. The patient underwent radical nephrectomy and tubular renal carcinoma was diagnosed histopathologically and immunohistochemically

Radiation-Induced Malignant Transformation of Preneoplastic and Normal Breast Primary Epithelial Cells.

Radiation is used in multiple procedures as a therapeutic and diagnostic tool. However, ionizing radiation can induce mutations in the DNA of irradiated cells, which can promote tumorigenesis. As malignant transformation is a process that takes many years, there are intermediate stages of cells that have initiated the process but have not yet evolved into cancer. The study here aimed to investigate the effect of ionizing radiation on normal and partially transformed human mammary epithelial cells. Breast primary epithelial cells were derived from normal breast tissue from two different donors and modified by transduction with the SV40 small and large T antigen and hTERT genes to obtain partially transformed cells and also with HRAS to completely and experimentally transform them. After exposure to different doses of ionizing radiation, oncogenic features were analyzed by means of an anchorage-independent growth assay and 3D cell culture. The addition of radiation exposure resulted in an increase in the number and size of colonies formed in each of the conditions analyzed and in the reduction of the capacity of partially transformed cells to form properly polarized 3D structures. Moreover, partially transformed cells require lower doses of radiation than healthy cells to enhance anchorage-independent growth capacity. Although cells from different donors have a different degree of sensitivity in the response to radiation, a higher sensitivity to the radiation-induced cell transformation process was observed in those cells that had already initiated the oncogenic process, which require higher doses of radiation to complete the transformation process. IMPLICATIONS: Individuals carrying accumulation of genetic alterations may have an increased susceptibility to radiation-induced neoplastic transformation.

Immunohistochemical expression of Ki-67, Cyclin D1, p16INK4a, and Survivin as a predictive tool for recurrence and progression-free survival in papillary urothelial bladder cancer pTa / pT1 G2 (WHO 1973).

OBJECTIVES: To investigate the expression of several immunohistochemical (IHC) markers and their predictive ability for the recurrence-free and progression-free survival of papillary urothelial bladder cancer (UBC) pTa/pT1 G2 (WHO 1973) compared to classical anatomo-clinical variables using a multidimensional analysis. MATERIALS AND METHODS: A population-based cohort of 213 primary stage UBC (pTa/pT1) G2 (WHO 1973) was evaluated by classic anatomopathological variables and characterized by immunohistochemistry (23 IHC markers, representative of different oncogenic pathways). The most important variables as a predictor of recurrence-free and progression-free survival were selected using multidimensional statistical models, such as random survival forests and least absolute shrinkage and selection operator (. Recurrence and progression-free survival of the previously selected variables were also calculated. RESULTS: Mean follow-up was 58 ± 33.5 months. Recurrence and progression rates were 54.5% (n = 116) and 17,4% (n = 37), respectively. The most influential variables in the low recurrence-free survival were in order: number of resected tumors, high expression of Ki67 (>10%), Cyclin D1 (>10%), and low cytoplasmic staining of p16INK4a. Regarding low progression-free survival, the most important variables were Ki67 (>15%), multicentric tumor arrangement and Survivin nuclear expression (>20%). Kaplan-Meier and cox-regression model analyses showed that the variables selected by multidimensional models were able to discriminate the clinical outcome. CONCLUSIONS: Ki67 index is the most useful IHC marker, since it can improve the prediction of both recurrence and progression-free survival in papillary UBC pTa/pT1 G2 (WHO 1973). There are other markers, whose utility is specific to recurrence-free survival, such as Cyclin D1 and p16INK4a or in progression-free survival, such as Survivin.