Cancer-Associated Fibroblasts Exert Proangiogenic Activity in Merkel Cell Carcinoma.

The tumor microenvironment is a complex niche enveloping a tumor formed by extracellular matrix, blood vessels, immune cells, and fibroblasts constantly interacting with cancer cells. Although tumor microenvironment is increasingly recognized as a major player in cancer initiation and progression in many tumor types, its involvement in Merkel cell carcinoma (MCC) pathogenesis is currently unknown. In this study, we provide a molecular and functional characterization of cancer-associated fibroblasts (CAFs), the major tumor microenvironment component, in patient-derived xenografts of patients with MCC. We show that subcutaneous coinjection of patient-derived CAFs and human MCC MKL-1 cells into severe combined immunodeficient mice significantly promotes tumor growth and metastasis. These fast-growing xenografts are characterized by areas densely populated with human CAFs, mainly localized around blood vessels. We provide evidence that the growth-promoting activity of MCC-derived CAFs is mediated by the aminopeptidase A/angiotensin II and III/angiotensin II type 1 receptor axis, with the expression of aminopeptidase A in CAFs being a triggering event. Together, our findings point to aminopeptidase A as a potential marker for MCC prognostic stratification and as a candidate for therapeutic intervention.

Merkel Cell Carcinoma and Immune Evasion: Merkel Cell Polyomavirus Small T-Antigen‒Induced Surface Changes Can Be Reverted by Therapeutic Intervention.

Merkel cell polyomavirus is the causative agent for most Merkel cell carcinomas (MCCs). This highly aggressive skin cancer shows rapid progression, with metastasis being a significant challenge for patient therapy. Virus-positive MCCs show low mutation rates, and tumor cell proliferation is dependent on viral oncoproteins small T antigen (sT) and large T antigen. Although the role of sT and large T antigen in early events of tumorigenesis has been extensively studied, their role in tumor progression has been scarcely addressed. In this study, we investigate the possible mechanisms of how Merkel cell polyomavirus oncoproteins, particularly sTs, contribute to metastasis. We show that sT specifically affects selectin ligand binding and processing by altering the presentation of multiple MCC surface molecules, thereby influencing initial metastasis events and tumor cell immune recognition. Furthermore, we show that sT regulates the surface antigen CD47, which inhibits phagocytosis by macrophages. By applying either sT short hairpin RNAs, CD47-targeted small interfering RNAs, or a therapeutic anti-CD47 antibody, we show that immune recognition of MCC cells can be restored. Thus, CD47 is a promising therapeutic target on MCC cells. Blocking the CD47‒SIRPα interaction effectively promotes phagocytosis of MCC cells and might be a promising combinatorial immunotherapy approach together with PD-1/PD-L1 axis in MCC treatment.

Ângulo de fase como potencial marcador do estado nutricional de pacientes em terapia intensiva / Phase angle as a potential marker of nutritional status of intensive care patients / El ángulo de fase como posible marcador del estado nutricional de los pacientes de cuidados intensivos

RESUMO Objetivo: verificar associação do ângulo de fase com estado nutricional, tempo de internação e óbito de pacientes críticos. Métodos: estudo longitudinal com 57 pacientes críticos avaliados na admissão em unidade de terapia intensiva durante o ano de 2019, no interior de São Paulo - BR. O ângulo de fase foi obtido por bioimpedância elétrica, e a avaliação nutricional, pelo instrumento de avaliação subjetiva global e antropometria. Os dados foram associados ao tempo de internação e óbito. Para a análise, utilizaram-se testes de Mann-Whitney e Qui-quadrado de Pearson. Resultados: na avaliação subjetiva global, 59,6% dos pacientes apresentavam risco nutricional, e em 91,2% o ângulo de fase estava baixo. A desnutrição foi associada ao maior tempo de internação hospitalar (p=0,001) em unidade de terapia intensiva (p =0,023). Verificou-se tendência ao óbito no grupo com risco nutricional (p=0,054). Conclusão: o ângulo de fase pode contribuir para melhor acurácia da avaliação nutricional, principalmente, quando combinado com outros métodos de avaliação.

ABSTRACT Objective: to verify the association of phase angle with nutritional status, length of hospitalization and death in critically ill patients. Methods: longitudinal study with 57 critically ill patients evaluated upon admission to an intensive care unit during the year 2019, in the countryside of São Paulo - BR. The phase angle was obtained by electrical bioimpedance, and nutritional assessment, by the global subjective assessment instrument and anthropometry. The data were associated with the time of hospitalization and death. For the analysis, Mann-Whitney and Pearson's chi-square tests were used. Results: in the global subjective evaluation, 59.6% of the patients presented nutritional risk, and in 91.2% the phase angle was low. Malnutrition was associated with longer hospital stay (p=0.001) in intensive care unit (p=0.023). There was a tendency to death in the group with nutritional risk (p=0.054). Conclusion: The phase angle can contribute to a better accuracy in nutritional assessment, especially when combined with other assessment methods.

RESUMEN Objetivo: Verificar la asociación del ángulo de fase con el estado nutricional, la duración de la estancia hospitalaria y la muerte en pacientes críticos. Métodos: estudio longitudinal con 57 pacientes críticos evaluados al ingreso en la unidad de cuidados intensivos durante el año 2019, en el interior de São Paulo - BR. El ángulo de fase se obtuvo mediante bioimpedancia eléctrica, y la valoración nutricional, mediante el instrumento de valoración subjetiva global y la antropometría. Los datos se asociaron al momento de la hospitalización y a la muerte. Para el análisis se utilizaron las pruebas de Mann-Whitney y Chi-cuadrado de Pearson. Resultados: en la evaluación subjetiva global, el 59,6% de los pacientes presentó riesgo nutricional, y en el 91,2% el ángulo de fase fue bajo. La desnutrición se asoció a una mayor estancia hospitalaria (p=0,001) en la unidad de cuidados intensivos (p=0,023). Hubo una tendencia a la muerte en el grupo con riesgo nutricional (p=0,054). Conclusión: el ángulo de fase puede contribuir a una mayor precisión en la evaluación nutricional, especialmente cuando se combina con otros métodos de evaluación.

Recorrência da internação por infecção do trato urinário em idosos / Recurrence of hospitalization for urinary tract infection in elderly / Recurrencia de la hospitalización por infección del tracto urinario en ancianos

Objetivos: Identificar, descrever e analisar a Infecção do Trato Urinário e sua recorrência em internação. Caracterizar os aspectos sociodemográficos, laboratoriais e clínicos dos idosos com ITU. Método: Estudo transversal com delineamento descritivo, abordagem quantitativa do tipo analítica, realizada nos prontuários eletrônicos dos idosos que foram internados no ano de 2017 e reinternaram em 2018, na unidade de Geriatria de um hospital de ensino. Na estatística foi usado Teste Exato de Fisher, considerado p<0,05. Resultados: Dos 1.835 prontuários, foram selecionados 550 (29,97%) idosos, destes, foram investigados 121 (22%) com ITU. A maioria foi do sexo masculino com idade entre 80 e 89 anos. Dentre eles, 51,2% usaram algum tipo de sonda vesical e 52,9% usaram frauda. Os exames alterados foram os de creatinina, Proteina-C-Reativa, urina I (proteínas e leucócitos), além de urocultura positiva para Escherichia Coli. Dos 121 pacientes investigados, 50,4% tiveram reinternação por Infecção urinária, destes, 42,6% morreram e, dos que não tiveram recorrência, foram 18,3% óbitos (p= 0,00054).Conclusão: O idoso tem predisposição maior para adquirir infeção urinária, além de apresentar recidivas, principalmente os que fazem uso de sonda vesical e fralda, o que ocasiona aumento da frequência de hospitalizações, do estadiamento no hospital e da mortalidade. (AU)

Objective: Analyze the Urinary Tract Infection and its recurrence in hospitalization in the elderly, characterizing the sociodemographic, laboratory and clinical aspects. Methods: Cross sectional study with descriptive design, quantitative approach of an analytical type, carried out on the electronic medical records of the elderly who were hospitalized in 2017 and readmitted in 2018, in the Geriatrics unit of a teaching hospital. Fisher's exact test was used in statistics, considered p <0.05. Results: Of the 1,835 records, 550 (29.97%) elderly were selected, of these, 121 (22%) with Urinary Tract Infection were investigated. The majority were male aged between 80 and 89 years. Among them, 51.2% used some type of bladder catheter and 52.9% used fraud. The altered tests were those of creatinine, C-Reactive Protein, urine I (proteins and leukocytes), in addition to positive urine culture for Escherichia Coli. Of the 121 patients investigated, 50.4% had readmission for urinary tract infection, of these, 42.6% died and, of those who did not have recurrence, 18.3% died (p = 0.00054). Conclusion: The elderly is predisposed greater to acquire urinary infection, in addition to relapses, especially those who use a bladder catheter and diaper, which causes an increase in the frequency of hospitalizations, hospital stay and mortality. (AU)

Objetivo: Analizar la Infección del Tracto Urinario y su recurrencia en la hospitalización del anciano, caracterizando los aspectos sociodemográficos, de laboratorio y clínicos. Métodos: Estudio transversal con diseño descriptivo, abordaje cuantitativo de tipo analítico, realizado sobre la historia clínica electrónica de los ancianos que fueron hospitalizados en 2017 y readmitidos en 2018, en la unidad de Geriatría de un hospital universitario. En estadística se utilizó la prueba exacta de Fisher, considerada p <0,05. Resultados: De las 1.835 historias clínicas, se seleccionaron 550 (29,97%) ancianos, de estos se investigaron 121 (22%) con Infección del Tracto Urinario. La mayoría eran hombres de entre 80 y 89 años. Entre ellos, el 51,2% utilizó algún tipo de sonda vesical y el 52,9% utilizó fraude. Las pruebas alteradas fueron las de creatinina, proteína C reactiva, orina I (proteínas y leucocitos), además de urocultivo positivo para Escherichia Coli. De los 121 pacientes investigados, el 50,4% tuvo reingreso por infección del tracto urinario, de estos, el 42,6% falleció y, de los que no presentaron recidiva, falleció el 18,3% (p = 0,00054). Conclusión: Los ancianos tienen mayor predisposición a adquirir urinaria infección, además de recaídas, especialmente en quienes utilizan sonda vesical y pañal, lo que provoca un aumento en la frecuencia de hospitalizaciones, estancia hospitalaria y mortalidad. (AU)

Evidence of BK Polyomavirus Infection in Urothelial but not Renal Tumors from a Single Center Cohort of Kidney Transplant Recipients.

Emerging evidence indicates that reactivation of BK polyomavirus (BKPyV) in the kidney and urothelial tract of kidney transplant recipients (KTRs) may be associated with cancer in these sites. In this retrospective study of a single center cohort of KTRs (n = 1307), 10 clear cell renal cell carcinomas and 5 urinary bladder carcinomas were analyzed from 15 KTRs for the presence of BKPyV infection through immunohistochemistry and fluorescent in situ hybridization (FISH). Three of these patients had already exhibited biopsy-proven polyomavirus-associated nephropathies (PyVAN). Although the presence of BKPyV large-T antigen was evident in the urothelium from a kidney removed soon after PyVAN diagnosis, it was undetectable in all the formalin-fixed and paraffin-embedded (FFPE) blocks obtained from the 10 kidney tumors. By contrast, large-T antigen (LT) labeling of tumor cells was detected in two out of five bladder carcinomas. Lastly, the proportion of BKPyV DNA-FISH-positive bladder carcinoma nuclei was much lower than that of LT-positive cells. Taken together, our findings further strengthen the association between BKPyV reactivation and cancer development in KTRs, especially bladder carcinoma.

Subversion of Host Innate Immunity by Human Papillomavirus Oncoproteins.

The growth of human papillomavirus (HPV)-transformed cells depends on the ability of the viral oncoproteins E6 and E7, especially those from high-risk HPV16/18, to manipulate the signaling pathways involved in cell proliferation, cell death, and innate immunity. Emerging evidence indicates that E6/E7 inhibition reactivates the host innate immune response, reversing what until then was an unresponsive cellular state suitable for viral persistence and tumorigenesis. Given that the disruption of distinct mechanisms of immune evasion is an attractive strategy for cancer therapy, the race is on to gain a better understanding of E6/E7-induced immune escape and cancer progression. Here, we review recent literature on the interplay between E6/E7 and the innate immune signaling pathways cGAS/STING/TBK1, RIG-I/MAVS/TBK1, and Toll-like receptors (TLRs). The overall emerging picture is that E6 and E7 have evolved broad-spectrum mechanisms allowing for the simultaneous depletion of multiple rather than single innate immunity effectors. The cGAS/STING/TBK1 pathway appears to be the most heavily impacted, whereas the RIG-I/MAVS/TBK1, still partially functional in HPV-transformed cells, can be activated by the powerful RIG-I agonist M8, triggering the massive production of type I and III interferons (IFNs), which potentiates chemotherapy-mediated cell killing. Overall, the identification of novel therapeutic targets to restore the innate immune response in HPV-transformed cells could transform the way HPV-associated cancers are treated.

Una serie de casos de pacientes con heridas crónicas después de recibir tratamiento tópico con plata / Series of cases of patients with chronic wounds after receiving topical treatment with silver

Muchos son los desafíos hallados en el cuidado de las heridas cutáneas, que alcanzan una proporción aún mayor cuando estas heridas presentan señales clínicas surgidas de infecciones. En gran parte de las instituciones nacionales de asistencia a la salud no es posible identificar y cuantificar los microrganismos presentes en las heridas. OBJETIVO: Identificar los microorganismos de las heridas crónicas antes y después del tratamiento con una cobertura antimicrobiana con plata y verificar los niveles de proteína C reactiva y su asociación con las dimensiones de la herida. Casuística y método: Serie de casos con análisis clínicos y de laboratorio de 10 pacientes con heridas crónicas, durante un período de 2 semanas en un ambulatorio de heridas del Hospital de Base/FUNFARME, de la Facultad de Medicina de São José do Rio Preto. RESULTADOS: De los pacientes estudiados, la mayoría (60%) eran del sexo femenino y un 40% presentó úlcera vasculogénica. El microorganismo predominante fue Staphylococcus aureus. Hubo inhibición de algunos microrganismos y crecimiento de otros géneros y especies. Se consideró débil la correlación entre el área inicial de la herida y la proteína C reactiva (PCR) sérica inicial (0,05183), y entre área final de la herida y la PCR sérica final (0,3587). Sin embargo, con el aumento del área de la herida hubo un incremento de los niveles séricos de PCR y, con la disminución del área, um reducción de los niveles de PCR. CONCLUSIÓN: La correlación entre los niveles séricos de PCR y las dimensiones de la herida fueron débiles y no significativas, pero positivas

Many challenges can be found on the care of cutaneous wounds, especially when these wounds present subtle clinical signs of infection. It is not possible to identify and quantify microorganisms in wounds in most national health care institutions. OBJECTIVES: To identify the microorganisms of chronic wounds before and after treatment with antimicrobial coverage with silver, and to check the levels of C-reactive protein and its association with the dimensions of the wound. Casuistic and METHOD: Case series with clinical and laboratory analysis of 10 patients with chronic wounds, during a period of two weeks in a Wound Care Outpatient Clinic / FUNFARME, Medical School of São José do Rio Preto / SP-FAMERP. RESULTS: Of the patients studied, the majority (60%) were female, and 40% had vasculogenic ulcer. The predominant microorganism was Staphylococcus aureus. There was inhibition of some microorganisms and growth of other genera and species. We found a weak correlation between initial wound area and initial serum CRP (0,05183), and between final wound area and final serum CRP (0,3587). However, with the increase of the area of the wound, there was an increase in the serum levels of the CRP and, with the decrease of the area, a reduction of the CRP levels. CONCLUSION: The correlation of serum CRP levels and wound dimensions were weak and not significant; but positive

Human Papillomavirus E7 Oncoprotein Subverts Host Innate Immunity via SUV39H1-Mediated Epigenetic Silencing of Immune Sensor Genes.

Subversion of innate immunity by oncoviruses, such as human papillomavirus (HPV), favors carcinogenesis because the mechanism(s) of viral immune evasion can also hamper cancer immunosurveillance. Previously, we demonstrated that high-risk (hr) HPVs trigger simultaneous epigenetic silencing of multiple effectors of innate immunity to promote viral persistence. Here, we expand on those observations and show that the HPV E7 oncoprotein upregulates the H3K9-specific methyltransferase, whose action shuts down the host innate immune response. Specifically, we demonstrate that SUV39H1 contributes to chromatin repression at the promoter regions of the viral nucleic acid sensors RIG-I and cGAS and the adaptor molecule STING in HPV-transformed cells. Inhibition of SUV39H1 leads to transcriptional activation of these genes, especially RIG-I, followed by increased beta interferon (IFN-ß) and IFN-λ1 production after poly(dA·dT) or RIG-I agonist M8 transfection. Collectively, our findings provide new evidence that the E7 oncoprotein plays a central role in dampening host innate immunity and raise the possibility that targeting the downstream effector SUV39H1 or the RIG-I pathway is a viable strategy to treat viral and neoplastic disease.IMPORTANCE High-risk HPVs are major viral human carcinogens responsible for approximately 5% of all human cancers. The growth of HPV-transformed cells depends on the ability of viral oncoproteins to manipulate a variety of cellular circuits, including those involved in innate immunity. Here, we show that one of these strategies relies on E7-mediated transcriptional activation of the chromatin repressor SUV39H1, which then promotes epigenetic silencing of RIG-I, cGAS, and STING genes, thereby shutting down interferon secretion in HPV-transformed cells. Pharmacological or genetic inhibition of SUV39H1 restored the innate response in HPV-transformed cells, mostly through activation of RIG-I signaling. We also show that IFN production upon transfection of poly(dA·dT) or the RIG-I agonist M8 predominantly occurs through RIG-I signaling. Altogether, the reversible nature of the modifications associated with E7-mediated SUV39H1 upregulation provides a rationale for the design of novel anticancer and antiviral therapies targeting these molecules.

Primary trichodysplasia spinulosa polyomavirus infection in a kidney transplant child displaying virus-infected decoy cells in the urine.

We report a case of primary trichodysplasia spinulosa (TS) infection in a kidney transplant child and describe for the first time the presence of degenerated TS-associated polyomavirus (TSPyV)-infected cells in a TS patient's urine that are morphologically different from BK or JC polyomavirus-infected decoy cells.

Association between diagnosis, clinical outcome and hospitalization time of the elderly in a university hospital / Associação entre diagnóstico, desfecho clínico e tempo de internação de idosos em um hospital escola

Clinical outcomes of hospitalized elderly provide subsidies to prevent complications that interfere with quality of life.The goal was to verify the sociodemographic characteristics and to analyze the associations between the diseasesdiagnosed at admission, age, hospitalization time and clinical outcome of elderly hospitalized in SUS and private care.Descriptive and quantitative cross-sectional survey of 14,892 medical records of elderly patients hospitalized in theyear 2014. The statistical tests used were chi-squared and analysis of variance. The results were: 93.5% white, 80.2%catholic, 68% had a primary education, 62.9% had partners, 53.8% males, 33.1% homemakers and the average agewas 72.1 years ± 8.6 years. The mean duration of hospitalization was 5.9 days ± 8.8 days, cardiovascular diseaseswere the most frequent, with a clinical outcome of 93.4% of hospital discharges and 6.6% of deaths, and 100% ofhematological diseases were discharged. There was a significant association rate between diseases and death frominfections (n=817; 60.0%). The association of diseases with the type of SUS care was statistically significant. Moreover,the occurrence of pain, nephro-urological diseases, orthopedic diseases, hematological and gynecological diseasesand other diseases showed a statistically significant association with private care or health plan. The associationbetween the diseases and the clinical outcome showed a greater incidence of death in patients affected by infections.The association of diseases was statistically significant with the type of care, highlighting the SUS care

Os desfechos clínicos de idosos internados proporcionam subsídios para prevenir complicações que interferem naqualidade de vida. O objetivo no estudo foi verificar as características sociodemográficas e analisar as associaçõesentre as doenças diagnosticadas na admissão, idade, tempo de internação e desfecho clínico de idosos internadosno SUS e convênio. Pesquisa transversal, descritiva e quantitativa, nos prontuários eletrônicos de 14.892 idososhospitalizados no ano de 2014. Os testes realizados foram Qui-quadrado e análise de variância. Constatou-se que93,5% eram brancos, 80,2% católicos, 68% possuíam ensino fundamental, 62,9% tinham companheiros, 53,8%homens, 33,1% do lar e a média de idade foram de 72,1 anos ± 8,6 anos. A média de tempo de internação foide 5,9 dias ± 8,8 dias, as doenças cardiovasculares foram as de maior ocorrência, tendo como desfecho clínico93,4% de altas hospitalares e 6,6% de óbitos e doenças hematológicas 100% de alta. Houve taxa significativa naassociação entre doenças e óbito por infecções (n=817; 60,0%). Foi estatisticamente significante a associação dasdoenças com o tipo de atendimento pelo SUS. Já a ocorrência de dor, doenças nefrourológicas, doenças ortopédicas,doenças hematológicas e ginecológicas e outras doenças apresentaram associação estatisticamente significativa com oatendimento particular ou convênio. A associação entre as doenças e o desfecho clínico demonstrou maior incidênciade óbito em pacientes acometidos por infecções. Foi estatisticamente significante a associação das doenças com o tipode atendimento destacando-se o atendimento pelo SUS

Desnutrição e complicações clínicas em pacientes cirúrgicos com doença colorretal / Malnutrition and clinical outcomes in surgical patients with colorectal disease

ABSTRACT BACKGROUND: Malnutrition is a frequent condition among hospitalized patients and a factor of increased risk of postoperative complication. OBJECTIVE: This study aimed to evaluate the impact of malnutrition on phase angle (PA), body water distribution and clinical outcomes in surgical patients with colorectal disease. METHODS: This retrospective study was performed in a tertiary hospital with 40 patients admitted electively. In the preoperative evaluation, global subjective assessment and bioelectrical impedance analysis were performed to determine nutritional status, PA, extracellular water (ECW), intracellular water (ICW) and total body water (TBW). In postoperative evaluation, the length of hospital stay and severe complications, according to Clavien-Dindo classification, were determined. The optimal PA cutoff for malnutrition screening was determined by ROC curve analysis. RESULTS: Seventeen (42.5%) patients were diagnosed as malnourished and 23 (57.5%) as well-nourished according to global subjective assessment. Twelve (30.0%) patients developed severe complications. The malnourished group presented lower values of serum albumin (P=0.012), hematocrit (P=0.026) and PA (P=0.002); meanwhile, ECW/ICW (P=0.019) and ECW/TBW (P=0.047) were higher. Furthermore, 58.8% of malnourished patients developed severe postoperative complications compared to 8.7% of well-nourished. Malnutrition was independent predictor of severe postoperative complications (OR=15.00, IC: 2.63-85.68, P=0.002). The optimal PA cutoff obtained was 6.0º (AUC=0.82, P=0.001), yielding sensitivity, specificity, positive predictive value and negative predictive value of 76.5%, 87.0%, 81.3% and 83.4%, respectively. CONCLUSION: Malnutrition was an independent predictive factor for severe complications in patients underwent to elective major coloproctological surgery. Besides that, malnutrition was associated with lower PA values and greater ratio of ECW. The PA provided great accuracy in nutritional screening, implying a useful marker of malnutrition.

RESUMO CONTEXTO: A desnutrição é uma condição frequente entre pacientes hospitalizados e é um fator de risco para complicações pós-operatórias. OBJETIVO: Este estudo tem como objetivo avaliar o impacto da desnutrição sobre o ângulo de fase (AF), a distribuição de água corporal e complicações clínicas em pacientes cirúrgicos com doença colorretal. MÉTODOS: Trata-se de um estudo retrospectivo realizado em um hospital universitário terciário com 40 pacientes admitidos eletivamente. Na avaliação pré-operatória, foram realizadas a avaliação subjetiva global e análise de bioimpedância elétrica com a finalidade de determinarem o estado nutricional, AF, água extracelular (AEC), água intracelular (AIC) e água corporal total (ACT). Na avaliação pós-operatória, o tempo de internação hospitalar e a presença de complicações graves, segundo a classificação de Clavien-Dindo, foram determinados. O melhor ponto de corte do AF para o rastreamento de desnutrição foi obtido a partir da análise da curva ROC. RESULTADOS: Dezessete (42,5%) pacientes foram diagnosticados como desnutridos e 23 (57,5%), como bem nutridos de acordo com a avaliação subjetiva global. Doze (30,0%) pacientes desenvolveram complicações pós-operatórias graves. O grupo desnutrido apresentou menores valores de albumina sérica (P=0,012), hematócrito (P=0,026) e AF (P=0,002); enquanto que as relações de AEC/AIC (P=0,019) e AEC/ACT (P=0,047) estiveram elevadas. Além disso, 58,8% dos pacientes desnutridos desenvolveram complicações pós-operatórias graves em comparação a 8,7% dos pacientes bem nutridos. A desnutrição foi fator preditivo independente para o desenvolvimento de complicações pós-operatórias graves (OR=15,00, IC: 2,63-85,68; P=0,002). O melhor ponto de corte do AF obtido foi 6.0º (AUC=0,82; P=0,001) com sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo de 76,5%, 87,0%, 81,3% e 83,4%, respectivamente. CONCLUSÃO: A desnutrição foi fator preditivo para o desenvolvimento de complicações graves em pacientes submetidos à cirurgia eletiva coloproctológica de grande porte. Além disso, a desnutrição foi associada a menores valores de AF e maior proporção de AEC. O AF forneceu boa acurácia no rastreamento da desnutrição, sugerindo seu uso como potencial marcador de desnutrição.

HPV18 Persistence Impairs Basal and DNA Ligand-Mediated IFN-ß and IFN-λ1 Production through Transcriptional Repression of Multiple Downstream Effectors of Pattern Recognition Receptor Signaling.

Although it is clear that high-risk human papillomaviruses (HPVs) can selectively infect keratinocytes and persist in the host, it still remains to be unequivocally determined whether they can escape antiviral innate immunity by interfering with pattern recognition receptor (PRR) signaling. In this study, we have assessed the innate immune response in monolayer and organotypic raft cultures of NIKS cells harboring multiple copies of episomal HPV18 (NIKSmcHPV18), which fully recapitulates the persistent state of infection. We show for the first time, to our knowledge, that NIKSmcHPV18, as well as HeLa cells (a cervical carcinoma-derived cell line harboring integrated HPV18 DNA), display marked downregulation of several PRRs, as well as other PRR downstream effectors, such as the adaptor protein stimulator of IFN genes and the transcription factors IRF1 and 7. Importantly, we provide evidence that downregulation of stimulator of IFN genes, cyclic GMP-AMP synthase, and retinoic acid-inducible gene I mRNA levels occurs at the transcriptional level through a novel epigenetic silencing mechanism, as documented by the accumulation of repressive heterochromatin markers seen at the promoter region of these genes. Furthermore, stimulation of NIKSmcHPV18 cells with salmon sperm DNA or poly(deoxyadenylic-deoxythymidylic) acid, two potent inducers of PRR signaling, only partially restored PRR protein expression. Accordingly, the production of IFN-ß and IFN-λ1 was significantly reduced in comparison with the parental NIKS cells, indicating that HPV18 exerts its immunosuppressive activity through downregulation of PRR signaling. Altogether, our findings indicate that high-risk human papillomaviruses have evolved broad-spectrum mechanisms that allow simultaneous depletion of multiple effectors of the innate immunity network, thereby creating an unreactive cellular milieu suitable for viral persistence.

MALNUTRITION AND CLINICAL OUTCOMES IN SURGICAL PATIENTS WITH COLORECTAL DISEASE.

BACKGROUND: Malnutrition is a frequent condition among hospitalized patients and a factor of increased risk of postoperative complication. OBJECTIVE: This study aimed to evaluate the impact of malnutrition on phase angle (PA), body water distribution and clinical outcomes in surgical patients with colorectal disease. METHODS: This retrospective study was performed in a tertiary hospital with 40 patients admitted electively. In the preoperative evaluation, global subjective assessment and bioelectrical impedance analysis were performed to determine nutritional status, PA, extracellular water (ECW), intracellular water (ICW) and total body water (TBW). In postoperative evaluation, the length of hospital stay and severe complications, according to Clavien-Dindo classification, were determined. The optimal PA cutoff for malnutrition screening was determined by ROC curve analysis. RESULTS: Seventeen (42.5%) patients were diagnosed as malnourished and 23 (57.5%) as well-nourished according to global subjective assessment. Twelve (30.0%) patients developed severe complications. The malnourished group presented lower values of serum albumin (P=0.012), hematocrit (P=0.026) and PA (P=0.002); meanwhile, ECW/ICW (P=0.019) and ECW/TBW (P=0.047) were higher. Furthermore, 58.8% of malnourished patients developed severe postoperative complications compared to 8.7% of well-nourished. Malnutrition was independent predictor of severe postoperative complications (OR=15.00, IC: 2.63-85.68, P=0.002). The optimal PA cutoff obtained was 6.0º (AUC=0.82, P=0.001), yielding sensitivity, specificity, positive predictive value and negative predictive value of 76.5%, 87.0%, 81.3% and 83.4%, respectively. CONCLUSION: Malnutrition was an independent predictive factor for severe complications in patients underwent to elective major coloproctological surgery. Besides that, malnutrition was associated with lower PA values and greater ratio of ECW. The PA provided great accuracy in nutritional screening, implying a useful marker of malnutrition.

The Nuclear DNA Sensor IFI16 Acts as a Restriction Factor for Human Papillomavirus Replication through Epigenetic Modifications of the Viral Promoters.

UNLABELLED: The human interferon-inducible IFI16 protein, an innate immune sensor of intracellular DNA, was recently demonstrated to act as a restriction factor for human cytomegalovirus (HCMV) and herpes simplex virus 1 (HSV-1) infection by inhibiting both viral-DNA replication and transcription. Through the use of two distinct cellular models, this study provides strong evidence in support of the notion that IFI16 can also restrict human papillomavirus 18 (HPV18) replication. In the first model, an immortalized keratinocyte cell line (NIKS) was used, in which the IFI16 protein was knocked down through the use of small interfering RNA (siRNA) technology and overexpressed following transduction with the adenovirus IFI16 (AdVIFI16) vector. The second model consisted of U2OS cells transfected by electroporation with HPV18 minicircles. In differentiated IFI16-silenced NIKS-HPV18 cells, viral-load values were significantly increased compared with differentiated control cells. Consistent with this, IFI16 overexpression severely impaired HPV18 replication in both NIKS and U2OS cells, thus confirming its antiviral restriction activity. In addition to the inhibition of viral replication, IFI16 was also able to reduce viral transcription, as demonstrated by viral-gene expression analysis in U2OS cells carrying episomal HPV18 minicircles and HeLa cells. We also provide evidence that IFI16 promotes the addition of heterochromatin marks and the reduction of euchromatin marks on viral chromatin at both early and late promoters, thus reducing both viral replication and transcription. Altogether, these results argue that IFI16 restricts chromatinized HPV DNA through epigenetic modifications and plays a broad surveillance role against viral DNA in the nucleus that is not restricted to herpesviruses. IMPORTANCE: Intrinsic immunity is mediated by cellular restriction factors that are constitutively expressed and active even before a pathogen enters the cell. The host nuclear factor IFI16 acts as a sensor of foreign DNA and an antiviral restriction factor, as recently demonstrated by our group for human cytomegalovirus (HCMV) and herpes simplex virus 1 (HSV-1). Here, we provide the first evidence that IFI16 inhibits HPV18 replication by repressing viral-gene expression and replication. This antiviral restriction activity was observed in immortalized keratinocytes transfected with the religated genomes and in U2OS cells transfected with HPV18 minicircles, suggesting that it is not cell type specific. We also show that IFI16 promotes the assembly of heterochromatin on HPV DNA. These changes in viral chromatin structure lead to the generation of a repressive state at both early and late HPV18 promoters, thus implicating the protein in the epigenetic regulation of HPV gene expression and replication.

O papel da Glumatina na terapia nutricional do transplante de medula óssea / The role of glumatine in nutritional support of bone marrow transplantation

A glutamina (L-GLN) é um aminoácido que localiza-se preferencialmente no músculo esquelético e é condicionalmente essencial nas situaçöes onde ocorre hipercatabolismo, como nos transplantes de medula óssea. Nestas situaçöes a suplementaçäo com a L-GLN na terapia nutricional é segura e recomendada. O emprego do aminoácido com objetivo de reduzir os efeitos secundários no TMO como mucosite e manifestaçöes digestivas parece existir. Existem dados que sugerem um efeito profilático da L-GLN em relaçäo à doença veno-oclusiva hepática nos pacientes transplantados. O emprego do aminoácido em combinaçäo com anti-oxidantes, o uso do mesmo via enteral e/ou parenteral, säo respostas que devem ser obtidas através de estudos em grupos homogêneos e selecionados de pacientes submetidos ao transplante de medula óssea.