Use of morphological, biochemical and SRAP molecular markers to differentiate varieties of Cynara cardunculus L. (Asteraceae)

Dentro del complejo primario de Cynara cardunculus L. se encuentran diversas variedades botánicas: var. scolymus (alcaucil), var. altilis (cardo cultivado) y var. sylvestris (cardo silvestre). A lo largo de la historia del mejoramiento, la caracterización de los materiales ha evolucionado desde el uso de caracteres morfológicos hasta los modernos análisis moleculares, pasando por los marcadores bioquímicos. El objetivo del presente trabajo fue comparar la utilidad de los marcadores morfológicos, bioquímicos y moleculares para la caracterización de materiales pertenecientes a tres variedades botánicas de Cynara cardunculus. Tres cultivares de la var. scolymus, dos de la var. altilis y dos de la var. sylvestris fueron caracterizadas por variables morfovegetativas, proteínas de reserva y por marcadores moleculares a través de la técnica SRAP. Estas metodologías permitieron discriminar dos grupos: uno incluyendo las variedades de cardo cultivado y silvestre, y el otro, las variedades del alcaucil. Los datos moleculares y morfológicos permitieron además diferenciar los cultivares evaluados de la var. scolymus. Se concluye que los marcadores analizados son útiles para la caracterización intravarietal e intervarietal en programas de mejoramiento.In the Cynara cardunculus L. primary complex we can find a globe artichoke: Cynara cardunculus var. scolymus and other botanical varieties such as C. cardunculus var. altilis (cultivated cardoon) and C. cardunculus var. sylvestris (wild artichoke). Traditionally, they varieties were grouped according to morphological and biochemical traits and more recently, based on molecular markers. The aim of the present paper was to compare the use of morphological, biochemical and molecular markers to distinguish botanical varieties of Cynara cardunculus. Three accessions of var. scolymus, two accessions of var. altilis and two accessions of var. sylvestris were used. Different morphological variables, electrophoresis seed protein patterns by SDS-PAGE and SRAP markers were evaluated. The three kinds of markers grouped all materials in two clusters: one of them included the globe artichoke and the other the cardoons. SDS-PAGE is a valid technique for botanical varieties identification but the morphological and molecular data were more effective at discriminating botanical varieties and accessions within the scolymus group.Fil: Espósito, Andrea. Consejo Nacional de Investigaciones Científicas y TécnicasFil: Martin, Eugenia. Consejo Nacional de Investigaciones Científicas y TécnicasFil: Cravero, Vanina P.. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias. Genética y Mejoramiento Vegetal.Fil: Cointry, Enrique L.. Consejo Nacional de Investigaciones Científicas y Técnica

Musculoskeletal Diseases Role in the Frailty Syndrome: A Case-Control Study.

Frailty syndrome severely burdens older age, and musculoskeletal diseases are of paramount importance in its development. The aim of this study is to unravel the contribution of musculoskeletal diseases to frailty syndrome. This is a case-control study, and we enrolled 55 robust community-dwelling age- and gender-matched patients, with 58 frail and pre-frail subjects. Frailty was diagnosed according to the Fried criteria (FP), and the Fragility Index (FI) was calculated. In all the subjects, a comprehensive geriatric assessment was carried out. Their nutritional status was evaluated by the Mini Nutritional Assessment and Bioelectrical Impedance Analyses. Their bone density (BMD), bone turnover, muscle mass, strength and performance were evaluated. Here, we show that the prevalence of frailty varies according to the diagnostic criteria used and that FP and FI showed a moderate to good agreement. Despite age and gender matching, frail subjects had lower muscle strength, performance and BMD. Their quality of life and cognitive performance were reduced in the frail subjects compared to the robust ones. Muscular strength and performance, together with mood, significantly predicted the diagnosis of frailty, whereas BMD and bone turnover did not. In conclusion, we show that sarcopenia plays a pivotal role in predicting the diagnosis of frailty, whereas osteoporosis does not

Role of liquid biopsy in oncogene-addicted non-small cell lung cancer

none7noThe discovery of actionable oncogene in non-small cell lung cancer (NSCLC) allowed the identification of a subgroup of patients who benefit from targeted tyrosine kinase inhibitors more than others. Mutations in the epidermal growth factor receptor (EGFR), translocations in the anaplastic lymphoma kinase (ALK) and rearrangements in the ROS proto-oncogene 1 (ROS1) must be identified in tumor tissue to guide the proper treatment choice. Liquid biopsy is based on the analysis of tumor materials released in the circulation. Liquid biopsy can be complementary to tissue biopsy, both at baseline and at progression, especially in the detection of somatic gene alterations emerging during the treatment with tyrosine kinase inhibitors (TKIs). Particularly, circulating DNA is used to find mutations in driver oncogenes, while circulating tumor cells, extracellular vesicles (EVs) and cell-free microRNAs (cfmiRNAs) are still under investigation. To help the unbiased use of liquid biopsy in the choice of the appropriate therapy, some recommendations were delivered by expert panels. Currently, analysis of EGFR mutations in cell-free DNA (cfDNA) is recommended at baseline when tissue biopsy harbors scarce tumor cells, and at progression before performing tissue biopsy; liquid biopsy analysis for other oncogenic drivers is not indicated in the clinical practice.openCanale M.; Pasini L.; Bronte G.; Delmonte A.; Cravero P.; Crino L.; Ulivi P.Canale, M.; Pasini, L.; Bronte, G.; Delmonte, A.; Cravero, P.; Crino, L.; Ulivi, P

New generation anaplastic lymphoma kinase inhibitors

Anaplastic lymphoma kinase (ALK) gene translocations are pro-tumoral driver alterations that encompass 3*7% of non-squamous non-small cell lung cancer (NSCLC) with specific, clinic and histologic features. The therapeutic strategy depends on anti-ALK tyrosine kinase inhibitors (TKIs) of which crizotinib was the first approved for clinical use. Despite its use improved significantly progression-free survival, overall response rate and duration of response of this illness, after a median period of 10.9 months all patients progress due to the development of acquired resistance mutations in the ALK tyrosine kinase domain in approximately one third of patients. Moreover, 60-90% of patients treated with crizotinib has a progression in the central nervous system (CNS) in absence of extracranial worsening of the disease. This is primarily attributed to poor CNS penetration by crizotinib as many pre-clinical and clinical models suggest. For instance, in order to overtake acquired resistance to crizotinib, prolong the control of the disease and manage CNS localizations, several II and III generation TKIs have been developed. Some of them were approved after the failure of crizotinib (ceritinib, alectinib, brigatinib and lorlatinib) and in first line setting (ceritinib, alectinib and brigatinib) while others are still under evaluation for TKI-naive patients such as lorlatinib, ensartinib and entrectinib. In this review we will discuss the most recent results of new TKIs in order to describe a fast growing therapeutic landscape in this setting

Statistical analysis of geochemical data: a tool for discriminating between kaolin deposits of hypogene and supergene origin, Patagonia, Argentina

The numerous kaolin deposits located in Patagonia, Argentina, have been formed by hypogene or supergene processes. The primary origin has been established from O18 and D isotopic composition of the main minerals, kaolinite and/or dickite, and from the behaviour of certain elements during the alteration. The aim of this paper was to find if there is a tool, other than oxygendeuterium data, to establish the origin of the Patagonian kaolin deposits. To handle the large number of variables per sample, a statistical multivariate study was used. The Principal Component method defines, on one hand the variables that better characterize each deposit and, on the other hand, the correlation between them. Fifty seven elements were considered and those that were not explained using these two components (which represent 75% of the total variance of the model) were discarded. As a result, the contents of Fe2O3, P2O5, LOI, Sr, Y, Zr, V, Pb, Hf, Rb, S and REE were used and the results show that the two components separate the deposits into two fields that are consistent with the process of formation. The first component indicates that Fe2O3, Y, Rb, U and HREE are more abundant in the supergene deposits, whereas, Sr, Pb, S and V are more abundant in the hypogene deposits. The second component shows that S, P2O5 and the LREE are enriched in the hydrothermal deposits, whereas Zr is more abundant in those formed under weathering conditions

Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina

Bovine babesiosis is a tick-borne disease caused by protozoan parasites of the genus Babesia. Babesia bigemina is one of the most prevalent and economically important parasite species that infects cattle because of its impact on the meat and milk production industry. Effective disease control strategies should include detection of reservoir animals and early and specific pathogen detection using rapid, economical, sensitive, and specific detection techniques. The loop-mediated isothermal amplification technique (LAMP) is a one-step molecular reaction that amplifies DNA sequences with high sensitivity and specificity under isothermal conditions and requires no special equipment. The results can be observed by the naked eye as color changes. The aim of this work was to develop and standardize the LAMP technique for B. bigemina detection and its visualization using hydroxynaphtol blue. For this situation, primers were designed from the conserved sequences of the B. bigemina ama-1 gene. The results showed that at 63 °C in 1 h and under standardized conditions, this technique could amplify B. bigemina DNA as indicated by the characteristic colorimetric change. Sensitivity evaluation indicated that DNA was amplified at a 0.00000001% parasitemia, and it was demonstrated that this technique specifically amplified the DNA of B. bigemina. Additionally, this technique could amplify DNA from 10 strains of B. bigemina from three different countries. It is concluded that the LAMP technique as modified in our case could specifically amplify B. bigemina DNA and shows high sensitivity, does not cross-react with related organisms, and the product is observed by 60 min of reaction time based on color changes. This report is the first LAMP report that uses sequences that are conserved between strains of the ama-1 gene, demonstrates the results by color changes using hydroxynaphtol blue. We propose LAMP as a rapid and economical alternative method for the molecular detection of B. bigemina.Fil: Lizarazo Zuluaga, Andrea P.. Universidad Autonoma de Queretaro.; MéxicoFil: Carvajal Gamez, Bertha I.. Universidad Autonoma de Queretaro.; MéxicoFil: Wilkowsky, Silvina Elizabeth. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Mosqueda, Juan. Universidad Autonoma de Queretaro.; Méxic

Post-Prior discrepancies in CDW-EIS calculations for ion impact ionization fully differential cross sections

In this work we present fully differential cross sections (FDCSs) calculations using post and prior version of CDW--EIS theory for helium single ionization by 100 MeV C$^{6+}$ amu$^{-1}$ and 3.6 MeV amu$^{-1}$ Au$^{24+}$ and Au$^{53+}$ ions. We performed our calculations for different momentum transfer and ejected electron energies. The influence of internuclear potential on the ejected electron spectra is taken into account in all cases. We compare our calculations with absolute experimental measurements. It is shown that prior version calculations give better agreement with experiments in almost all studied cases.Comment: 9 pages, 7 figure

Liquid biopsy for egfr mutation analysis in advanced non-small-cell lung cancer patients: Thoughts drawn from a real-life experience

none12noBackground: Liquid biopsy analysis for EGFR detection in cell-free DNA (cfDNA) from NSCLC patients has become routine. The aim of this study was to explore its applicability in clinical practice. Methods: We collected data of EGFR-mutated NSCLC patients with liquid biopsy analysis. Data included test timing, concomitant tissue re-biopsy, therapy change, histology, stage, smoking habits, gender and age. All analyses were performed via a real-time PCR method to analyze EGFR mutations at exons 18, 19, 20 and 21. Variant allele frequency was performed for patients with available sequential EGFR mutation analysis in cfDNA. Overall survival was analyzed through the Kaplan–Meier method. We designed flow charts to show the real-life application of liquid biopsy. Results: We found that liquid biopsy is used in treatment-naïve patients as an alternative to EGFR detection in tumor tissue, and in patients with positive or negative EGFR from tumor biopsy. The majority of liquid biopsy analyses were performed in NSCLC patients who were disease progressive during TKI therapy. The presence of EGFR mutation in cfDNA was associated with a worse prognosis. In two patients, VAF of EGFR mutations in cfDNA was concordant with tumor volume changes. Conclusion: These findings suggest that liquid biopsy for EGFR detection can continue to be useful.openUlivi P.; Petracci E.; Canale M.; Priano I.; Capelli L.; Calistri D.; Chiadini E.; Cravero P.; Rossi A.; Delmonte A.; Crino L.; Bronte G.Ulivi, P.; Petracci, E.; Canale, M.; Priano, I.; Capelli, L.; Calistri, D.; Chiadini, E.; Cravero, P.; Rossi, A.; Delmonte, A.; Crino, L.; Bronte, G

The Interplay Between Programmed Death Ligand 1 and Vimentin in Advanced Non-Small-Cell Lung Cancer

none14noBackground: Current therapy for non-small-cell lung cancer (NSCLC) frequently includes immune checkpoint inhibitors, such as pembrolizumab, and programmed death ligand 1 (PD-L1) positivity is mandatory for its use in this setting. Vimentin plays a role in carcinogenesis through the activation of the epithelial-to-mesenchymal transition (EMT) process. Its prognostic impact in NSCLC has been investigated in numerous studies but little data are available on its relation with PD-L1 expression. Patients and Methods: We retrospectively retrieved data on patients with advanced NSCLC consecutively enrolled in a clinical trial at our institute. PD-L1 and vimentin expression were determined by immunohistochemistry. Correlations between variables were assessed using the Spearman correlation coefficient. The Kaplan-Meier method was used to estimate overall survival (OS) and the Log-rank test was used to compare survival curves. The association between demographic, clinical and biomarker information and survival was investigated with the Cox model. Results: Fifty-three patients were included in the study. A weak positive correlation was observed between the PD-L1 and vimentin (ρ=0.41, P=0.003). Patients with PD-L1 values <1% showed a slightly better OS than those with higher values (HR=2.07; 95% CI: 0.92–4.65), but the difference was not significant (P=0.080). No difference in overall survival (OS) was observed on the basis of vimentin expression (HR=1.25; 95% CI: 0.59–2.66; P=0.554). Patients harboring both vimentin and PD-L1 negative expression (<1%) showed a trend towards better survival than those with ≥1% expression (HR=2.31; 95% CI: 0.87-6.17, P=0.093). No significant associations were observed between gender, age at diagnosis, stage at diagnosis, histology, KRAS or EGFR status, radical surgery or immunotherapy and OS. Conclusions: The weak positive association between PD-L1 and vimentin suggests a potential interplay between these biomarkers. Further research is warranted to evaluate EMT and immune escape as two components of the same process.openBronte G.; Puccetti M.; Petracci E.; Landi L.; Cravero P.; Scodes S.; Ulivi P.; Ravaioli S.; Tumedei M.M.; Burgio M.A.; Cappuzzo F.; Delmonte A.; Crino L.; Bravaccini S.Bronte, G.; Puccetti, M.; Petracci, E.; Landi, L.; Cravero, P.; Scodes, S.; Ulivi, P.; Ravaioli, S.; Tumedei, M. M.; Burgio, M. A.; Cappuzzo, F.; Delmonte, A.; Crino, L.; Bravaccini, S