Analysis of the interaction of calcitriol with the disulfide isomerase ERp57

Calcitriol, the active form of vitamin D3, can regulate the gene expression through the binding to the nuclear receptor VDR, but it can also display nongenomic actions, acting through a membrane- associated receptor, which has been discovered as the disul de isomerase ERp57. The aim of our research is to identify the binding sites for calcitriol in ERp57 and to analyze their interaction. We rst studied the interaction through bioinformatics and uorimetric analyses. Subsequently, we focused on two protein mutants containing the predicted interaction domains with calcitriol: abb’- ERp57, containing the rst three domains, and a’-ERp57, the fourth domain only. To consolidate the achievements we used the calorimetric approach to the whole protein and its mutants. Our results allow us to hypothesize that the interaction with the a’ domain contributes to a greater extent than the other potential binding sites to the dissociation constant, calculated as a Kd of about 10−9 M

Variation of the Nutritional Composition of Quinoa According to the Processing Used

ARTÍCULO PUBLICADO EN REVISTA EXTERNA. Quinoa is consumed as a seed, flour, expanded, sprout (germinated) and activated (hydrated). The objective of this work was to determine the nutritional composition of the different preparations. The same batch of quinoa seeds was processed as flour, expanded, hydrated and germinated. It showed that there is a statistically significant difference of nutrients between all groups. For proteins, it varies from 12.78 ± 0.02 g/100 g in whole seed to 5.25 ± 0.01 g/100 g in the hydrated seed. In total fats, it varies from 7.80 ± 0.02 g/100 g in flour to 0.72 ± 0.01 g/100 g in sprouts. For fiber, the germinated quinoa provides the highest content (23.50 ± 0.01 g/100 g), whereas the hydrated quinoa the lowest content (8.71 ± 0.02 g/100 g). This shows how different preparations influence the nutritional contribution of quinoa. With this information, one can recommend different types of preparations depending on the type of nutrient that is wanted for consumption

Variation of the nutritional composition of quinoa according to the processing used

PRE-PRINTS (VERSIÓN FINAL DE AUTOR PARA PUBLICACIÓN). In Mendoza, Argentina, quinoa is grown with a very good yield, and its consumption is becoming widespread. It is consumed as a seed, flour, expanded, sprout (germinated) and activated (hydrated). The popular belief is that all these forms have the same nutritional contribution. Taking into account this belief, the objective of this work was to determine the nutritional composition of the different preparations. The same batch of quinoa seeds was processed as flour, expanded, hydrated and sprout. It was analyzed by triplicate, applying conventional laboratory techniques. For the statistical analysis, a multiple comparison test was applied, and to discriminate between the means, the honestly significant difference procedure (HSD) of Tukey was applied. It showed that there is statistically significant difference of nutrients between all groups, with a level of 95.0% confidence. For proteins, it varies from 12.78±0.02 g/100g in whole seed to 5.25±0.01 g/100g in the hydrated seed. In total fats, it varies from 7.80±0.02 g/100g in flour to 0.72±0.01 g/100g in sprouts. For fiber, the germinated quinoa provides the highest content (23.50±0.01 g/100g), whereas the hydrated quinoa the lowest content (8.71±0.02 g/100 g). The energy value in 100 g was as follows: whole seed 1299 kcal, flour 1430 kcal, germinated 291 kcal, hydrated 594 kcal and expanded 1368 kcal. This shows how different preparations influence the nutritional contribution of quinoa. With this information, one can recommend different types of preparations depending on the type of nutrient which is wanted for consumption

Detección de hongos productores de micotoxinas en semillas de chía que se expenden en el comercio

En el año 2014, en Estados Unidos y en Canadá se observó intoxicación por Salmonella spp. en consumidores de chía orgánica. Esto motivo el interés de realizar el análisis microbiológico de las semillas que se expenden en Argentina, porque es frecuente el consumo de semillas directamente en crudo, las cuales se agregan a ensaladas, leche, yogur o se hidratan y se consumen como un gel, sin ningún tratamiento

Management of Chronic Myeloid Leukemia in Advanced Phase

Management of chronic myeloid leukemia (CML) in advanced phases remains a challenge also in the era of tyrosine kinase inhibitors (TKIs) treatment. Cytogenetic clonal evolution and development of resistant mutations represent crucial events that limit the benefit of subsequent therapies in these patients. CML is diagnosed in accelerated (AP) or blast phase (BP) in <5% of patients, and the availability of effective treatments for chronic phase (CP) has dramatically reduced progressions on therapy. Due to smaller number of patients, few randomized studies are available in this setting and evidences are limited. Nevertheless, three main scenarios may be drawn: (a) patients diagnosed in AP are at higher risk of failure as compared to CP patients, but if they achieve optimal responses with frontline TKI treatment their outcome may be similarly favorable; (b) patients diagnosed in BP may be treated with TKI alone or with TKI together with conventional chemotherapy regimens, and subsequent transplant decisions should rely on kinetics of response and individual transplant risk; (c) patients in CP progressing under TKI treatment represent the most challenging population and they should be treated with alternative TKI according to the mutational profile, optional chemotherapy in BP patients, and transplant should be considered in suitable cases after return to second CP. Due to lack of validated and reliable markers to predict blast crisis and the still unsatisfactory results of treatments in this setting, prevention of progression by careful selection of frontline treatment in CP and early treatment intensification in non-optimal responders remains the main goal. Personalized evaluation of response kinetics could help in identifying patients at risk for progression

A cholinergic-sympathetic pathway primes immunity in hypertension and mediates brain-to-spleen communication

The crucial role of the immune system in hypertension is now widely recognized. We previously reported that hypertensive challenges couple the nervous drive with immune system activation, but the physiological and molecular mechanisms of this connection are unknown. Here, we show that hypertensive challenges activate splenic sympathetic nerve discharge to prime immune response. More specifically, a vagus-splenic nerve drive, mediated by nicotinic cholinergic receptors, links the brain and spleen. The sympathetic discharge induced by hypertensive stimuli was absent in both coeliac vagotomized mice and in mice lacking α7nAChR, a receptor typically expressed by peripheral ganglionic neurons. This cholinergic-sympathetic pathway is necessary for T cell activation and egression on hypertensive challenges. In addition, we show that selectively thermoablating the splenic nerve prevents T cell egression and protects against hypertension. This novel experimental procedure for selective splenic denervation suggests new clinical strategies for resistant hypertension

The contribution of vascular and extra-vascular water pathways to drought-induced decline of leaf hydraulic conductance

Drought stress can impair leaf hydraulic conductance (Kleaf), but the relative contribution of changes in the efficiency of the vein xylem water pathway and in the mesophyll route outside the xylem in driving the decline of Kleaf is still debated. We report direct measurements of dehydration-induced changes in the hydraulic resistance (R=1/K) of whole leaf (Rleaf), as well as of the leaf xylem (Rx) and extra-vascular pathways (Rox) in four Angiosperm species. Rleaf, Rx, and Rox were measured using the vacuum chamber method (VCM). Rleaf values during progressive leaf dehydration were also validated with measurements performed using the rehydration kinetic method (RKM). We analysed correlations between changes in Rx or Rox and Rleaf, as well as between morpho-anatomical traits (including dehydration-induced leaf shrinkage), vulnerability to embolism, and leaf water relation parameters. Measurements revealed that the relative contribution of vascular and extra-vascular hydraulic properties in driving Kleaf decline during dehydration is species-specific. Whilst in two study species the progressive impairment of both vascular and extra-vascular pathways contributed to leaf hydraulic vulnerability, in the other two species the vascular pathway remained substantially unaltered during leaf dehydration, and Kleaf decline was apparently caused only by changes in the hydraulic properties of the extra-vascular compartment

Validation of a Liquid Biopsy Protocol for Canine BRAFV595E Variant Detection in Dog Urine and Its Evaluation as a Diagnostic Test Complementary to Cytology

A significant proportion of canine urothelial carcinomas carry the driver valine to glutamic acid variation (V595E) in BRAF kinase. The detection of V595E may prove suitable to guide molecularly targeted therapies and support non-invasive diagnosis of the urogenital system by means of a liquid biopsy approach using urine. Three cohorts and a control group were included in this multi-step validation study which included setting up a digital PCR assay. This was followed by investigation of preanalytical factors and two alternative PCR techniques on a liquid biopsy protocol. Finally, a blind study using urine as diagnostic sample has been carried out to verify its suitability as diagnostic test to complement cytology. The digital PCR (dPCR) assay proved consistently specific, sensitive, and linear. Using the dPCR assay, the prevalence of V595E in 22 urothelial carcinomas was 90.9%. When compared with histopathology as gold standard in the blind-label cases, the diagnostic accuracy of using the canine BRAF (cBRAF) variation as a surrogate assay against the histologic diagnosis was 85.7% with 92.3% positive predictive value and 80.0% negative predictive value. In all the cases, in which both biopsy tissue and the associated urine were assayed, the findings matched completely. Finally, when combined with urine sediment cytology examination in blind-label cases with clinical suspicion of malignancy, the dPCR assay significantly improved the overall diagnostic accuracy. A liquid biopsy approach on urine using the digital PCR may be a valuable breakthrough in the diagnostic of urothelial carcinomas in dogs

Neoangiogenesis markers in canine urothelial carcinomas: A cross‐sectional study

Background: In humans, there is a growing body of evidence that neoangiogenesis is crucial for tumour growth and progression in urothelial carcinomas (UC) which also typically exhibit overactivation of the RAS-MAPK pathway. In canine UC (cUC), the same pathway has been aberrantly activated due to V595E BRAF variant and BRAF inhibitors has been evaluated as more effective treatment. However, BRAF inhibition is hampered in humans by rapidly occurring of chemoresistance. Targeting angiogenesis has been speculated to increase the effectiveness of BRAF inhibitors and to delay the development of chemoresistance. Objectives: This study aimed to investigate the level of angiogenic markers in urine samples of UC affected dogs (n = 15) in comparison to an unmatched control group (n = 16) along with the clinical, morphological and molecular features. Methods: In urine, both vascular endothelial growth factor (VEGF) concentration, using an ELISA assay, and MMP2 and 9 activities, using the zymographic assay, were measured. BRAF analysis was carried out using a digital PCR method. Results: Urinary VEGF concentration (mean pg/g_uCrea 6.9 +/- 27.7 vs. 1074 +/- 1797, p < 0.01) and MMP activity (mean 6.8 x 10(6) +/- 9.2 x 10(6) vs. 2.5 x 10(7) +/- 2.3 x 10(7), p < 0.05) were higher in affected dogs than in healthy controls. Urinary active MMP9 was significantly correlated with T3 stage, it was absent in dogs with undetectable VEGF and it correlated well with urinary VEGF concentration. In this cohort, 10/10 UC affected dogs exhibited the V595E BRAF variation. Conclusion: The findings are consistent with the presence of overactive neoangiogenesis in cUC. Urinary active MMP9 may be suitable for use as tumour progression biomarker. The addition of angiogenesis targeting may be rationale for novel therapeutic strategies

Tailored treatment including radical prostatectomy and radiation therapy + androgen deprivation therapy versus exclusive radical prostatectomy in high-risk prostate cancer patients: results from a prospective study

Purpose To evaluate outcomes of patients with high risk prostate cancer (PCa) who underwent radical prostatectomy (RP) in a context of a multidisciplinary approach including adjuvant radiation (RT) + androgen deprivation therapy (ADT). Matherials and Methods 244 consecutive patients with high risk localized PCa underwent RP and bilateral extended pelvic lymph node dissection at our institution. Adjuvant RT + 24 months ADT was carried out in subjects with pathological stage ≥ T3N0 and/or positive surgical margins or in patients with local relapse. Results After a median follow-up was 54.17 months (range 5.4-117.16), 13 (5.3%) subjects had biochemical progression, 21 (8.6%) had clinical progression, 7 (2.9%) died due to prostate cancer and 15 (6.1%) died due to other causes. 136 (55.7%) patients did not receive any adjuvant treatment while 108 (44.3%) received respectively adjuvant or salvage RT+ADT. Multivariate Cox proportional hazard analysis showed that pre-operative PSA value at diagnosis is a significant predictive factor for BCR (HR: 1.04, p < 0.05) and that Gleason Score 8-10 (HR: 2.4; p<0.05) and PSMs (HR: 2.01; p < 0.01) were significant predictors for clinical progression. Radical prostatectomy group was associated with BPFS, CPFS, CSS and OS at 5-years of 97%, 90%, 95% and 86% respectively, while adjuvant radiation + androgen deprivation therapy group was associated with a BPFS, CPFS and CSS at 5-years of 91%, 83%, 95% and 88%, without any statistical difference. Conclusions Multimodality tailored treatment based on RP and adjuvant therapy with RT+ADT achieve similar results in terms of OS after 5-years of follow-up