Regulation of natural killer cell cytotoxic pathways during serial killing activity

Natural Killer (NK) cells act as the front line of the body´s defense in the immune system and they are key players in efficiently recognizing and eliminating virus-infected and tumor cells. Individual NK cells can eliminate multiple target cells in a sequential manner during a process named serial killing. There are two major pathways NK cells use to induce apoptosis in their target cells: By releasing the content of cytotoxic granules containing the serine protease granzyme B (GrzB) and the pore-forming protein perforin or by the engagement of death receptors, which initiates caspase cascades via Caspase-8 (Casp8). The contribution of both cell death processes to NK cell cytotoxicity and serial killing remains poorly understood. Therefore, investigating the interplay between these pathways in more detail is important for a better understanding of NK cell cytotoxicity. To visualize the GrzB and death receptor-mediated target cell death in a time-dependent manner, we used fluorescent localization reporters that enabled us to simultaneously measure the activities of GrzB and of Casp8 in target cells upon contact with NK cells by life cell imaging. In this study we observed that NK cells kill their initial targets via the fast GrzB-induced pathway and switch to a slow death receptor-mediated killing for the final target. During the target cell contact NK cells lost GrzB and perforin, whereas the expression of CD95L, a main death cell ligand, was increased on the NK cell surface. The reduction of the lytic granules can be efficiently restored by the stimulation with different cytokines such as IL- 15, IL-2 or IL-21. Perforin deficient NK cells or ILC3 cells were unable to perform GrzB-mediated killing and no serial killing could be mediated without this pathway. In contrast, the absence of death receptor CD95 on the target cell and/or the absence of death receptor ligands on the NK cell had no direct influence on the GrzB-mediated serial killing. This demonstrates that the use of GrzB vs. death receptor-mediated target cell killing is differentially regulated during the serial killing activity of NK cells. Taken together, we observed a rapid target cell death which was induced by GrzB and originated from early established NK : target contacts. In contrast, cell death mediated by Casp-8 was a result of later target cell engagements and took much longer from NK : target cell contact to target cell death

Intertextuality in the Legal Papers of Karl Kraus: A Scholarly Digital Edition

Projektberich

Efecto del fósforo y potasio en la producción de ácido cítrico utilizando una cepa de aspergillus niger

En la investigación se evaluaron el rendimiento (%) y productividad (g/L por hora) de la fermentación sumergida de ácido cítrico con el uso de una cepa de Aspergillus niger mejorada por exposición a luz UV, cuando se incrementa el nivel de potasio (K) y reduce el nivel de fósforo (P) en una formulación de base que contiene sacarosa, micro- y macronutrientes. Se evaluaron cinco tratamientos en un diseño completamente al azar con tres repeticiones en una estructura multifactorial completo aumentado 22 + punto central. Los mejores resultados de rendimiento y productividad se obtuvieron con 329 mg/lt de P y 271 mg/L de K (tratamiento 2), equivalentes a 63.2% de rendimiento y 1.20 g/L por hora de productividad. Se encontró que sólo el potasio es altamente significativo y que no existe interacción significativa entre K y P, adicionalmente los resultados obtenidos podrían generar una reducción en los costos de producción de ácido cítrico. A partir del análisis de regresión de superficie de respuesta se obtuvieron modelos útiles para predecir los resultados de productividad y rendimiento de la fermentación de ácido cítrico dentro del rango de evaluación. Igualmente se encontró que el 8.01% de la variación total para la productividad y el 48.94% de la variación total para el rendimiento no se explican con el modelo. Los resultados obtenidos son diferentes a los reportados por otros autores debido a que estos han utilizado condiciones sub-óptimas para asegurar la máxima acumulación de ácido cítrico y/o cepas de colección las cuales se caracterizan por una menor productividad en ácido cítrico en comparación con las cepas industriales (Papa-gianni, 2007)

Prognostic significance of macrophage invasion in hilar cholangiocarcinoma

Background: Tumor-associated macrophages (TAMs) promote tumor progression and have an effect on survival in human cancer. However, little is known regarding their influence on tumor progression and prognosis in human hilar cholangiocarcinoma. Methods: We analyzed surgically resected tumor specimens of hilar cholangiocarcinoma (n = 47) for distribution and localization of TAMs, as defined by expression of CD68. Abundance of TAMs was correlated with clinicopathologic characteristics, tumor recurrence and patients’ survival. Statistical analysis was performed using SPSS software. Results: Patients with high density of TAMs in tumor invasive front (TIF) showed significantly higher local and overall tumor recurrence (both ρ < 0.05). Furthermore, high density of TAMs was associated with decreased overall (one-year 83.6 % vs. 75.1 %; three-year 61.3 % vs. 42.4 %; both ρ < 0.05) and recurrence-free survival (one-year 93.9 % vs. 57.4 %; three-year 59.8 % vs. 26.2 %; both ρ < 0.05). TAMs in TIF and tumor recurrence, were confirmed as the only independent prognostic variables in the multivariate survival analysis (all ρ < 0.05). Conclusions: Overall survival and recurrence free survival of patients with hilar cholangiocarcinoma significantly improved in patients with low levels of TAMs in the area of TIF, when compared to those with a high density of TAMs. These observations suggest their utilization as valuable prognostic markers in routine histopathologic evaluation, and might indicate future therapeutic approaches by targeting TAMs

Germline HOXB13 mutations p.G84E and p.R217C do not confer an increased breast cancer risk

In breast cancer, high levels of homeobox protein Hox-B13 (HOXB13) have been associated with disease progression of ER-positive breast cancer patients and resistance to tamoxifen treatment. Since HOXB13 p.G84E is a prostate cancer risk allele, we evaluated the association between HOXB13 germline mutations and breast cancer risk in a previous study consisting of 3,270 familial non-BRCA1/2 breast cancer cases and 2,327 controls from the Netherlands. Although both recurrent HOXB13 mutations p.G84E and p.R217C were not associated with breast cancer risk, the risk estimation for p.R217C was not very precise. To provide more conclusive evidence regarding the role of HOXB13 in breast cancer susceptibility, we here evaluated the association between HOXB13 mutations and increased breast cancer risk within 81 studies of the international Breast Cancer Association Consortium containing 68,521 invasive breast cancer patients and 54,865 controls. Both HOXB13 p.G84E and p.R217C did not associate with the development of breast cancer in European women, neither in the overall analysis (OR = 1.035, 95% CI = 0.859-1.246, P = 0.718 and OR = 0.798, 95% CI = 0.482-1.322, P = 0.381 respectively), nor in specific high-risk subgroups or breast cancer subtypes. Thus, although involved in breast cancer progression, HOXB13 is not a material breast cancer susceptibility gene.Peer reviewe

Atrasentan and renal events in patients with type 2 diabetes and chronic kidney disease (SONAR): a double-blind, randomised, placebo-controlled trial

Background: Short-term treatment for people with type 2 diabetes using a low dose of the selective endothelin A receptor antagonist atrasentan reduces albuminuria without causing significant sodium retention. We report the long-term effects of treatment with atrasentan on major renal outcomes. Methods: We did this double-blind, randomised, placebo-controlled trial at 689 sites in 41 countries. We enrolled adults aged 18–85 years with type 2 diabetes, estimated glomerular filtration rate (eGFR)25–75 mL/min per 1·73 m 2 of body surface area, and a urine albumin-to-creatinine ratio (UACR)of 300–5000 mg/g who had received maximum labelled or tolerated renin–angiotensin system inhibition for at least 4 weeks. Participants were given atrasentan 0·75 mg orally daily during an enrichment period before random group assignment. Those with a UACR decrease of at least 30% with no substantial fluid retention during the enrichment period (responders)were included in the double-blind treatment period. Responders were randomly assigned to receive either atrasentan 0·75 mg orally daily or placebo. All patients and investigators were masked to treatment assignment. The primary endpoint was a composite of doubling of serum creatinine (sustained for ≥30 days)or end-stage kidney disease (eGFR <15 mL/min per 1·73 m 2 sustained for ≥90 days, chronic dialysis for ≥90 days, kidney transplantation, or death from kidney failure)in the intention-to-treat population of all responders. Safety was assessed in all patients who received at least one dose of their assigned study treatment. The study is registered with ClinicalTrials.gov, number NCT01858532. Findings: Between May 17, 2013, and July 13, 2017, 11 087 patients were screened; 5117 entered the enrichment period, and 4711 completed the enrichment period. Of these, 2648 patients were responders and were randomly assigned to the atrasentan group (n=1325)or placebo group (n=1323). Median follow-up was 2·2 years (IQR 1·4–2·9). 79 (6·0%)of 1325 patients in the atrasentan group and 105 (7·9%)of 1323 in the placebo group had a primary composite renal endpoint event (hazard ratio [HR]0·65 [95% CI 0·49–0·88]; p=0·0047). Fluid retention and anaemia adverse events, which have been previously attributed to endothelin receptor antagonists, were more frequent in the atrasentan group than in the placebo group. Hospital admission for heart failure occurred in 47 (3·5%)of 1325 patients in the atrasentan group and 34 (2·6%)of 1323 patients in the placebo group (HR 1·33 [95% CI 0·85–2·07]; p=0·208). 58 (4·4%)patients in the atrasentan group and 52 (3·9%)in the placebo group died (HR 1·09 [95% CI 0·75–1·59]; p=0·65). Interpretation: Atrasentan reduced the risk of renal events in patients with diabetes and chronic kidney disease who were selected to optimise efficacy and safety. These data support a potential role for selective endothelin receptor antagonists in protecting renal function in patients with type 2 diabetes at high risk of developing end-stage kidney disease. Funding: AbbVie

Germline HOXB13 mutations p.G84E and p.R217C do not confer an increased breast cancer risk

Abstract: In breast cancer, high levels of homeobox protein Hox-B13 (HOXB13) have been associated with disease progression of ER-positive breast cancer patients and resistance to tamoxifen treatment. Since HOXB13 p.G84E is a prostate cancer risk allele, we evaluated the association between HOXB13 germline mutations and breast cancer risk in a previous study consisting of 3,270 familial non-BRCA1/2 breast cancer cases and 2,327 controls from the Netherlands. Although both recurrent HOXB13 mutations p.G84E and p.R217C were not associated with breast cancer risk, the risk estimation for p.R217C was not very precise. To provide more conclusive evidence regarding the role of HOXB13 in breast cancer susceptibility, we here evaluated the association between HOXB13 mutations and increased breast cancer risk within 81 studies of the international Breast Cancer Association Consortium containing 68,521 invasive breast cancer patients and 54,865 controls. Both HOXB13 p.G84E and p.R217C did not associate with the development of breast cancer in European women, neither in the overall analysis (OR = 1.035, 95% CI = 0.859–1.246, P = 0.718 and OR = 0.798, 95% CI = 0.482–1.322, P = 0.381 respectively), nor in specific high-risk subgroups or breast cancer subtypes. Thus, although involved in breast cancer progression, HOXB13 is not a material breast cancer susceptibility gene

Actividad de fosfatasas ácida y alcalina en suelo cultivado con plátano en tres sistemas de manejo

En suelo rizosférico de plátano (Musa AAB), bajo un diseño de bloques completos al azar, con tres sistemas de manejo (químico o convencional, tradicional y orgánico o agroecológico), cuatro profundidades (1-5, 5-10, 10-20 y 20-30 cm) y tres edades del cultivo (6 meses –diferenciación floral, 12 meses –floración, 18 meses –cosecha) se estimó la actividad de fosfatasas ácida y alcalina. El análisis estadístico arrojó diferencias significativas entre los tres manejos, presentándose mayor actividad de fosfatasa ácida en el tradicional y de fosfatasa alcalina en el manejo químico. Con res­pecto a la profundidad, para ambas enzimas la mayor actividad se encontró de 0-5 cm. En referencia a las edades del cultivo, la máxima actividad de fosfatasa ácida se presentó en la etapa de diferenciación floral (6 meses) y la máxima actividad de fosfatasa alcalina en la época de floración (12 meses) y cosecha (18 meses). Dado el carácter ácido del suelo estudiado (pH 5.1), la fosfatasa ácida presentó el 85% de la actividad y el restante 15% fue para fosfatasa alcalina. La actividad de fosfatasa ácida y alcalina está influenciada por el manejo agronómico y estado fisiológico del cultivo de plátano. El estudio de la dinámica enzimática permite visualizar cambios a corto plazo producidos por factores antropogénicos y asociados con producción de cultivos de plátano. Palabras claves: Actividad enzimática, fosfatasa ácida, fosfatasa alcalina, Andisol, Musa AAB.En suelo rizosférico de plátano (Musa AAB), bajo un diseño de bloques completos al azar, con tres sistemas de manejo (químico o convencional, tradicional y orgánico o agroecológico), cuatro profundidades (1-5, 5-10, 10-20 y 20-30 cm) y tres edades del cultivo (6 meses –diferenciación floral, 12 meses –floración, 18 meses –cosecha) se estimó la actividad de fosfatasas ácida y alcalina. El análisis estadístico arrojó diferencias significativas entre los tres manejos, presentándose mayor actividad de fosfatasa ácida en el tradicional y de fosfatasa alcalina en el manejo químico. Con res­pecto a la profundidad, para ambas enzimas la mayor actividad se encontró de 0-5 cm. En referencia a las edades del cultivo, la máxima actividad de fosfatasa ácida se presentó en la etapa de diferenciación floral (6 meses) y la máxima actividad de fosfatasa alcalina en la época de floración (12 meses) y cosecha (18 meses). Dado el carácter ácido del suelo estudiado (pH 5.1), la fosfatasa ácida presentó el 85% de la actividad y el restante 15% fue para fosfatasa alcalina. La actividad de fosfatasa ácida y alcalina está influenciada por el manejo agronómico y estado fisiológico del cultivo de plátano. El estudio de la dinámica enzimática permite visualizar cambios a corto plazo producidos por factores antropogénicos y asociados con producción de cultivos de plátano. Palabras claves: Actividad enzimática, fosfatasa ácida, fosfatasa alcalina, Andisol, Musa AAB