Real-time lipid patterns to classify viable and necrotic liver tumors.

Real-time tissue classifiers based on molecular patterns are emerging tools for fast tumor diagnosis. Here, we used rapid evaporative ionization mass spectrometry (REIMS) and multivariate statistical analysis (principal component analysis-linear discriminant analysis) to classify tissues with subsequent comparison to gold standard histopathology. We explored whether REIMS lipid patterns can identify human liver tumors and improve the rapid characterization of their underlying metabolic features. REIMS-based classification of liver parenchyma (LP), hepatocellular carcinoma (HCC), and metastatic adenocarcinoma (MAC) reached an accuracy of 98.3%. Lipid patterns of LP were more similar to those of HCC than to those of MAC and allowed clear distinction between primary and metastatic liver tumors. HCC lipid patterns were more heterogeneous than those of MAC, which is consistent with the variation seen in the histopathological phenotype. A common ceramide pattern discriminated necrotic from viable tumor in MAC with 92.9% accuracy and in other human tumors. Targeted analysis of ceramide and related sphingolipid mass features in necrotic tissues may provide a new classification of tumor cell death based on metabolic shifts. Real-time lipid patterns may have a role in future clinical decision-making in cancer precision medicine.

Acute lead arsenate poisoning in beef cattle in Uruguay.

We describe and illustrate lesions in an outbreak of lead arsenate poisoning in beef cattle that ingested pesticide residues stored in an abandoned building of a former orange orchard. Of 70 exposed cattle, 14 had diarrhea, paresis, ataxia, recumbency, and/or seizures. Ten of the affected animals died after a clinical course of 12-18 h. Pathologic findings in 3 steers included extensive necrohemorrhagic, ulcerative rumenitis, omasitis, and abomasitis; lymphocytolysis in lymphoid organs; and nephrosis. Hepatic arsenic and lead levels in cases 1-3 were 20, 24, and 31 ppm, and 8.3, 25, and 9.4 ppm, respectively. Lesions in the forestomachs and lymphoid tissues have been rarely reported in cases of lead arsenate poisoning. In southern South America, these lesions are indistinguishable from those produced by Baccharis coridifolia, a toxic plant that contains macrocyclic trichothecenes, thus these conditions should be considered in the differential diagnosis of necrotizing lesions in alimentary and lymphoid organs.

Etiología de la mastitis crónica: propuesta de secuencia diagnóstica / Proposed diagnosis sequence for chronic mastitis aetiology

Introducción: Las lesiones inflamatorias mamarias precisan con frecuencia de estudio histopatológico por su capacidad de imitar a los tumores mamarios malignos. El objetivo es proponer una secuencia diagnóstica de las inflamaciones mamarias benignas crónicas. Material y método: Se han revisado en la literatura los métodos y algoritmos diagnósticos de las mastitis crónicas. Resultados: Se propone un algoritmo diagnóstico para los procesos inflamatorios crónicos mamarios. Requiere determinar el patrón histopatológico inflamatorio y su localización, así como un estudio microbiológico apropiado. Posteriormente puede precisar de nuevas pruebas bioquímicas y serológicas orientadas por una correlación clinicopatológica para establecer un diagnóstico específico. Discusión: No se han identificado en la literatura otros algoritmos diagnósticos avalados por estudios de alto nivel de evidencia. Los patrones histopatológicos no son uniformes. Conclusiones: El diagnóstico etiológico precisa identificar patrones histopatológicos inflamatorios benignos y su localización, un estudio microbiológico y pruebas orientadas por correlación clinicopatológica. Se precisan estudios de investigación con niveles de evidencia altos

Introduction: Inflammatory breast lesions require histopathological study due to their ability to clinically and radiologically mimic malignant mammary tumours. The objective is to propose a diagnostic technique for benign chronic inflammatory processes of the breast. Material and methods: We reviewed the literature on the diagnostic methods used in chronic mastitis. Results: We propose a diagnostic algorithm for chronic inflammatory processes of the breast. The aetiological diagnosis requires identifying benign inflammatory histopathologic patterns and locations, and microbiological study. New biochemical and serological tests oriented by clinicopathological correlation may then be required to establish a specific diagnosis. Discussion: No diagnostic algorithms based on studies with a high level of evidence have been identified. No uniformity in histopathologic patterns has been described. Conclusions: The etiologic diagnosis requires identifying benign inflammatory histopathologic patterns and locations, microbiological study and tests oriented by clinicopathological correlation. There is a lack of studies with a high level of evidence

Targeting of regulated necrosis in kidney disease.

The term acute tubular necrosis was thought to represent a misnomer derived from morphological studies of human necropsies and necrosis was thought to represent an unregulated passive form of cell death which was not amenable to therapeutic manipulation. Recent advances have improved our understanding of cell death in acute kidney injury. First, apoptosis results in cell loss, but does not trigger an inflammatory response. However, clumsy attempts at interfering with apoptosis (e.g. certain caspase inhibitors) may trigger necrosis and, thus, inflammation-mediated kidney injury. Second, and most revolutionary, the concept of regulated necrosis emerged. Several modalities of regulated necrosis were described, such as necroptosis, ferroptosis, pyroptosis and mitochondria permeability transition regulated necrosis. Similar to apoptosis, regulated necrosis is modulated by specific molecules that behave as therapeutic targets. Contrary to apoptosis, regulated necrosis may be extremely pro-inflammatory and, importantly for kidney transplantation, immunogenic. Furthermore, regulated necrosis may trigger synchronized necrosis, in which all cells within a given tubule die in a synchronized manner. We now review the different modalities of regulated necrosis, the evidence for a role in diverse forms of kidney injury and the new opportunities for therapeutic intervention.

Terapia fotodinâmica em células de tumores pancreáticos humanos: eficiência e análise das vias mediadoras de citotoxicidade / Photodynamic therapy in human pancreatic tumors: efficiency and analysis of cytotoxicity mediator pathways

O adenocarcinoma de ducto pancreático (PDAC) é a quarta causa de morte em decorrência de neoplasias nos países ocidentais. Atualmente, a cirurgia ressectiva é a única possibilidade de cura para a doença, porém, a recidiva tumoral acontece em menos de um ano após a intervenção cirúrgica, mesmo com a quimioterapia adjuvante. A terapia fotodinâmica (PDT) é uma alternativa promissora no tratamento do câncer. No entanto, pouco se sabe sobre o uso da PDT em tumores pancreáticos. Portanto, o objetivo deste trabalho foi avaliar a eficiência da PDT com o azul de metileno (MB) como fotossensibilizador (MB-PDT) em induzir a morte de linhagens de PDAC humanas (AsPC-1, Panc-1, MIAPaCa-2 e BxPC-3) e estudar a contribuição de vias de necrose regulada nos efeitos citotóxicos da terapia sobre estes modelos. Os resultados obtidos mostraram que a MB-PDT foi capaz de induzir a morte massiva das células de PDAC. Além disso, eles indicaram que há dois perfis de susceptibilidade entre as quatro linhagens estudadas quando submetidas a MBPDT com 4,5 J/cm2 de energia e 6min de irradiação. De acordo com os dados apresentados, a diferença nas sensibilidades das linhagens à terapia não está associada à diferenças na capacidade de incorporação do MB ou na localização sub-celular do fotossensibilizador nas diferentes células, uma vez que a localização é, predominantemente, lisossomal em todas elas. Adicionalmente, mostrou-se que as linhagens menos susceptíveis ao tratamento, MIAPaCa-2 e Panc-1, apresentam níveis significativamente menores de RIPK3 e MLKL, dois dos componentes do necrossomo, essenciais para a execução da necroptose. Além disso, foi visto que a MB-PDT induz um aumento de fosforilação de MLKL em AsPC-1, demonstrando a ativação da necroptose após a terapia nestas células, mas não em MIAPaCa-2 (menos responsiva à terapia com 4,5 J/cm2 deenergia e 6min de tempo de irradiação). Ainda, a inibição da via de sinalização necroptótica diminuiu significativamente as porcentagens de morte das células mais susceptíveis (BxPC-3 e AsPC-1), não alterando a resposta de Panc-1 e MIAPaCa-2, corroborando a ativação e importância da necroptose para a citotoxicidade da MB-PDT. Finalmente, neste trabalho foi mostrado que o aumento do tempo de irradiação, mantendo-se a quantidade total de energia aplicada no tratamento, melhora a eficiência da MB-PDT em induzir a morte das células que apresentam limitações para executar a necroptose, sugerindo que mais de uma via de morte esteja sendo ativada após a terapia e que o tempo de irradiação atuaria modulando esta ativação. Complementarmente, foi mostrado que os tempos maiores de irradiação aumentam o estresse oxidativo intracelular que é acompanhado por uma diminuição significativa do conteúdo intracelular de glutationa reduzida (GSH), indicando, preliminarmente, que a ferroptose pode estar sendo acionada após os protocolos mais longos de irradiação. Coletivamente, os resultados apresentados neste trabalho confirmam a eficiência da MB-PDT no tratamento de diferentes linhagens de PDAC, indicando que a necroptose está sendo ativada e contribuindo para a citotoxicidade da terapia sobre as células que não apresentam resistência à esta via de morte. Ainda, eles demonstram que o aumento do tempo de irradiação pode transpor a barreira de resistência de algumas linhagens à terapia, provavelmente por induzir a ativação de outras vias de necrose regulada, mostrando a importância da otimização do protocolo de tratamento no aumento da eficiência da MB-PDT sobre os tumores de pâncreas. Finalmente, os resultados confirmam a MB-PDT como alternativa eficaz no tratamento do PDAC, apresentando um amplo espectro de atuação sobre subtipos tumorais resistentes à vias clássicas de morte celular, uma característica importante no contexto de uma terapia anti-cancer

Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of death due to neoplasms in western countries. Currently, resective surgery is the only therapetical approach to cure this disease, but tumor´s recurrence occurs less than one year after the surgery, even with adjuvant chemotherapy. Photodynamic therapy (PDT) is a promising alternative for the cancer treatment. However, the efficacy of PDT to treat pancreatic tumors as well as the mechanisms involved in the induction of tumorigenic cell death remain unclear. For this purpose, in this study, we set out to evaluate the efficacy of PDT using methylene blue (MB) as a photosensitizer (MB-PDT), in inducing death of human PDAC derived cell lines (AsPC-1, Panc-1, MIAPaCa-2 and BxPC-3) and to deeper investigate the contribution of necroptosis to the cytotoxic effects of the therapy. We observed that MB-PDT was able to induce massive death of PDAC cells. Moreover, our results indicated that upon MB-PDT (4.5 J/cm2 energy and 6min of irradiation time), there were two susceptibility profiles among the four cell lines studied. Data also showed that this differential profile of cell response was neither associated with the differences in the MB incorporation capacity nor with the subcellular location of the photosensitizer, since the localization was predominantly lysosomal in all of tested cell lines. In addition, less susceptible cells, MIAPaCa-2 and Panc-1, showed significantly lower levels of RIPK3 and MLKL, two of the necrosome components, essential for triggering necroptosis. Furthermore, while MB-PDT (4.5 J/cm2 and 6min of irradiation) has been able to increase MLKL´s phosphorylation levels, an essential step in necroptosis induction, in AsPC-1cells, less responsive MIAPaCa-2 cells presented no variations on the phosphorylation state of this pseudokinase. Moreover, pharmacological inhibition of the necroptotic signaling pathway significantly decreased cell death percentages of the most susceptible cells (BxPC-3 andAsPC-1), without altering the response of Panc-1 and MIAPaCa-2, corroborating that activation of necroptosis was strongly involved in the cytotoxicity of MB-PDT. Finally, this work showed that increasing the irradiation time improved the efficacy of MB-PDT in killing cells which display limitations to perform necroptosis, suggesting that the irradiation time would be modulating the degree of oxidative stress generated and this stimuli would in turn, be responsible for triggering other regulated cell death pathways in a RIKP3 and MLKL independent way. Indeed, this increase in oxidative stress was accompanied by a significant decrease in GSH, a global indicatior of less antioxidant cell capacity, preliminarily pointing at the induction of ferroptosis by longer irradiation protocols. In summary, we demonstrated that MB-PDT is able to induce cell death in different PDAC cell lines and that different regulated cell death mechanisms are being activated upon MB-PDT induction. Furthermore, it was demonstrated that increased irradiation time may overcome the resistance barrier of some cell lines, probably inducing the activation of other regulated cell death pathways, showing the importance of optimizing the irradiation protocol in order to maximize the efficacy of the therapy. Finally, our observations point MB-PDT as an alternative and effective therapy for pancreatic cancer treatment, displaying a broad-spectrum action on tumors displaying different resistance mechanisms to classic cell death pathways, a desired property for improving an anticancer therapy

Columellar Reconstruction After Nasal Continuous Positive Airway Pressure Associated Necrosis.

Though a life-saving modality in neonatal intensive care units, nasal continuous positive airway pressure (nCPAP) carries a small risk of irreversible ischemia and necrosis of the columella due to the configuration of the pressure delivery system. Iatrogenic injuries to the columella after nCPAP use result in a spectrum of disfigurement and functional airway obstruction. The authors performed a retrospective review of patients evaluated for nCPAP-related columellar deformities by the Division of Plastic and Reconstructive Surgery at the authors' institution over a 10-year period to assess reconstructive outcomes. Of 7 patients evaluated, 3 underwent reconstruction using a combination of cartilaginous framework reshaping and local tissue flaps. After a mean follow-up period of 78 months, patients had satisfactory aesthetic and functional results. Based on the authors' observations, columellar necrosis secondary to nCPAP can be divided into 3 categories: Type A demonstrates mild notching of the columella; Type B has an absent columella without notable nasal tip depression; Type C has an absent columella with nasal tip depression, with or without external nasal valve obstruction. Reconstructive needs should be individually tailored based on the degree of nasal tip depression, cartilaginous support, and soft tissue availability.

Response to chemotherapy estimates by FDG PET is an important prognostic factor in patients with Ewing sarcoma

Background. Response to chemotherapy is a prognostic factor in patients with Ewing sarcoma (ES); the role of FDG PET to predict response in these patients has not been thoroughly investigated. We evaluated the diagnostic accuracy and the potential of FDG PET to predict response to chemotherapy (CHT). Materials and methods. e analyzed data of 50 patients with ES (median age 12.6 years). All patients were treated with neoadjuvant CHT, and underwent surgery for local control. All patients had 18F-FDG PET/CT at diagnosis and after induction CHT, prior to local control. We compared response assessed by histopathology with FDG PET using standard uptake values (SUVs). Results. Median SUV at diagnosis (SUV I) was 5 (range 1.2-17), and median SUV after neoadjuvant chemotherapy (SUV II) was 1.8 (range 0-8.4). Median SUV II/I ratio was 0.3 (range 0-1). SUV at diagnosis was significantly lower in patients with good histological response than in patients with poor histological response (median 3.8 vs. 7.2, p 0.02). We found a significant correlation between SUV II and outcome; the positive predictive value of an SUV II ≤ 2.5 for favorable response was 84.21 %, and the median SUV II was significantly higher in patients with disease progression (2.3 vs. 1.6, p = 0.04). In multivariate analysis, necrosis and SUV II were significant predictors of outcome. Conclusions. 18F-FDG PET demonstrates high diagnostic accuracy for response to initial chemotherapy in patients with ES and it correlates with outcome. The role of FDG PET in predicting response and outcome should be further investigated (AU)

No disponible

A novel bowel necrosis classification system and examination of patient outcomes in incarcerated groin hernia patients.

The objective of this study was to determine a classification system for BN in incarcerated groin hernia patients and to explore the possible relationship between BN staging and patient outcomes. Incarcerated groin hernia patients treated with emergency bowel resection from January 2008 to December 2013 were screened for inclusion in a prospective study. A novel three-stage classification system was proposed for BN (BN stages I-III) and correlations between adverse events (AEs) and mortality with BN stage were determined. A total of 108 patients were included, with 71, 26, and 11 patients in BN stages I, II, and III, respectively. AEs, which included wound and intra-abdominal infections and other systemic complications, increased with higher BN stage (all P < 0.05). Mortality increased with BN stage, with 2.8%, 7.7%, and 27.3% at BN stages I, II, and III, respectively (P < 0.05). The proposed BN staging system can objectively reflect the degree of bowel damage and its corresponding adverse outcomes.

Necroptosis: an emerging form of programmed cell death.

Necrosis plays an important role in multiple physiological and pathological processes. Recently, a relatively new form of necrosis has been characterized as "necroptosis". Morphologically, necroptosis exhibits the features of necrosis; however, necroptosis exhibits a unique signaling pathway that requires the involvement of receptor interaction protein kinase 1 and 3 (RIP1 and RIP3) and can be specifically inhibited by necrostatins. Necroptosis has been found to contribute to the regulation of immune system, cancer development as well as cellular responses to multiple stresses. In this review, we will summarize the signaling pathway, biological effects and pathological significance of this specific form of programmed cell death.

Necrotizing soft tissue infections in the intensive care unit.

Necrotizing soft tissue infection is a severe illness that is associated with significant morbidity and mortality. It is often caused by a wide spectrum of pathogens and is most frequently polymicrobial. Care for patients with necrotizing soft tissue infection requires a team approach with expertise from critical care, surgery, reconstructive surgery, and rehabilitation specialists. The early diagnosis of necrotizing soft tissue infection is challenging, but the keys to successful management of patients with necrotizing soft tissue infection are early recognition and complete surgical debridement. Early initiation of appropriate broad-spectrum antibiotic therapy must take into consideration the potential pathogens. Critical care management components such as the initial fluid resuscitation, end-organ support, pain management, nutrition support, and wound care are all important aspects of the care of patients with necrotizing soft tissue infection. Soft tissue reconstruction should take into account both functional and cosmetic outcome.

AIF-mediated programmed necrosis: a highly regulated way to die.

Historically, two main forms of cell death have been distinguished: apoptosis and necrosis. Apoptosis was initially considered as the only physiological and programmed form of cell death. This type of death is recurrently associated with caspases, a family of cysteine proteases activated in apoptotic conditions. However, it is now widely recognized that programmed cell death (PCD) can also occur in the complete absence of caspase activation. The existence of non-caspase PCD pathways was corroborated by the discovery of caspase-independent executioners, such as the mitochondrial protein Apoptosis-Inducing Factor (AIF). Necrosis has often been viewed as an accidental and uncontrolled cell death process. Nevertheless, increasing evidence shows that, like apoptosis, necrosis could be a highly orchestrated type of PCD. Indeed, apoptosis and necrosis present more similarities than it has been originally thought. Here, we summarize the different classifications of PCD and the current knowledge of a necrotic PCD pathway mediated by AIF: alkylating DNA-damage mediated death. We also outline the molecular mechanisms controlling this form of PCD and discuss their potential relevance in physiological and pathological settings. These emerging data on the molecular mechanisms regulating programmed necrosis may certainly have potent therapeutic consequences in treating both apoptotic-resistant tumors and degenerating adult neurons.

Oxigenoterapia hiperbárica para infecciones necrotizantes de partes blandas / Hyperbaric oxygenotherapy for necrotizing infections of soft parts

Introducción: La Oxigenoterapia Hiperbárica (OHB) se ha convertido en el tratamiento de elección de numerosas patologías. Sin embargo, su rol en el tratamiento de las Infecciones Necrotizantes de partes blandas (INPB) es aún controvertido.Objetivo: Evaluar el beneficio de la Oxigenoterapia Hiperbárica (OHB) en el tratamiento de las Infecciones Necrotizantes de partes blandas (INPB).Método: Retrospectivo, revisión de historias clínicas y seguimiento de los casos. Durante el período de Enero 1996 a Diciembre2002, una población de 42 pacientes con INPB, se categorizó las lesiones de acuerdo a profundidad según la clasificación de Ámsterdam y se los dividió en 2 grupos: el grupo I (n = 18) que completó el protocolo de 10 sesione de OHB de 60 minutos cada una a 2.5 atmósferas absolutas (ATA), iniciando el 1º día del postoperatorio y el grupo II (n = 24) que no lo completó. En todos los casos se realizó desbridamiento quirúrgico precoz y antibioticoterapia.Resultado: En el grupo I hubo 12 (66.6%) recuperaciones completas, 5 (27.7%) secuelas leves y 1 (5.7%) grave, en el grupo II hubo 5 (20.8%) recuperaciones completas, 8 (33.4%) secuelas leves y 5 (20.8%) graves. La flora patógena fue en el 85.7% de los casos polimicrobiana. La mortalidad global fue del 25%, todos del grupo II.Conclusión: Se observó una relación inversamente proporcional entre el número de sesiones de OHB y la morbimortalidad. La OHB fue beneficiosa en esta serie como tratamiento complementario a la cirugía y los antibióticos.

Análisis comparativo de la cirugía de la necrosis pancreática en las dos últimas décadas / Comparative study of surgery in pancreatic necrosis in the last 2 decades

Con la finalidad de examinar nuestros resultados en el tratamiento quirúrgico de la necrosis pancreática se analizan retrospectivamente 31 pacientes a los que se les practicó necrosectomía en un lapso de 20 años (1971-1990). En la serie predominó el sexo masculino (22 enfermos). Se detallan las características clínicas que, junto a las pruebas de laboratorio y diagnóstico por imágenes, permitieron sospechar la necrosis con infección y decidir la oportunidad quirúrgica. La mortalidad global postoperatoria fue de 11 pacientes (35,48 por ciento). En la primera década sobre 12 operados observamos una letalidad de 8 casos (66,66 por ciento), mientras que en el segundo decenio ésta se redujo a 3 observaciones (15,78 por ciento). La completa exéresis de los tejidos desvitalizados y el aporte de una correcta reanimación pre y postoperatoria en unidades de cuidado intensivo, sumados a una nutrición parenteral o enteral que permitió evitar una realimentación oral temprana, a un mejor manejo infectológico y a la incorporación de nuevos procedimientos de diagnóstico por imágenes constituyeron la base para mejorar los resultados en los últimos 19 casos de nuestra casuística

Análisis comparativo de la cirugía de la necrosis pancreática en las dos últimas décadas / Comparative study of surgery in pancreatic necrosis in the last 2 decades

Con la finalidad de examinar nuestros resultados en el tratamiento quirúrgico de la necrosis pancreática se analizan retrospectivamente 31 pacientes a los que se les practicó necrosectomía en un lapso de 20 años (1971-1990). En la serie predominó el sexo masculino (22 enfermos). Se detallan las características clínicas que, junto a las pruebas de laboratorio y diagnóstico por imágenes, permitieron sospechar la necrosis con infección y decidir la oportunidad quirúrgica. La mortalidad global postoperatoria fue de 11 pacientes (35,48 por ciento). En la primera década sobre 12 operados observamos una letalidad de 8 casos (66,66 por ciento), mientras que en el segundo decenio ésta se redujo a 3 observaciones (15,78 por ciento). La completa exéresis de los tejidos desvitalizados y el aporte de una correcta reanimación pre y postoperatoria en unidades de cuidado intensivo, sumados a una nutrición parenteral o enteral que permitió evitar una realimentación oral temprana, a un mejor manejo infectológico y a la incorporación de nuevos procedimientos de diagnóstico por imágenes constituyeron la base para mejorar los resultados en los últimos 19 casos de nuestra casuística (AU)

Necrotizing fasciitis: a classification of necrotizing soft tissue infections.

Necrotizing fasciitis is a rare, often fatal soft tissue infection. It still remains a confusing entity because of the nomenclature and multiple subtypes described in the past. An interesting case study of a patient with necrotizing fasciitis secondary to nonclostridial gas gangrene is presented. A comprehensive review of necrotizing fasciitis, its disease process and treatment modalities will be discussed.

Loxosceles y loxoscelismo en Piura / Loxosceles and loxoscelism in Piura

Los autores presentan dos casos con loxocelismo cutaneo y mordedura de araña del género Loxoceles sp. en la casa de uno de los pacientes. Estos son uno de los primeros caos de loxocelismo y Loxoceles en Piura

[Necrotizing vasculitis with predominant kidney involvement]. / Vasculitis necrotizante con predominio de afección renal.

We report five patients with necrotizing vasculitis with predominant renal involvement; the diagnostic and therapeutics was effected completely at regional hospital. We detach: a) the clinical presentation's forms with rapidly progressive renal failure with microscopic hematuria and minimal proteinuria; b) the biopsy of kidney with necrotizing vasculitis or necrotizing glomerulonephritis (microscopic polyarteritis). We review the necrotizing vasculitis with predominant renal involvement in its pathogenic, clinical and histological aspects, and we insist that the diagnostic and early treatment are fundamental for prognostic's improvement of these patients. We conclude that this diagnostic and treatment can be made in regional hospitals with sensible physicians by the theme.