Contrasting metabolic profiles of tasty Andean varieties of tomato fruit in comparison with commercial ones

BACKGROUND The fruits of most commercial tomato cultivars (Solanum lycopersicum L.) are deficient in flavour. In contrast, traditional ‘criollo’ tomato varieties are appreciated for fruit of excellent organoleptic quality. Small farmers from the Andean valleys in Argentina have maintained their own tomato varieties, which were selected mainly for flavour. This work aims to correlate the chemical composition of the fruit with the sensory attributes of eight heirloom tomato varieties. The long‐term goal is to identify potential candidate genes capable of altering the chemicals involved in flavour. RESULTS A sensory analysis was conducted and the metabolomics of fruit were determined. The data revealed that defined tomato aroma and sourness correlated with citrate and several volatile organic compounds (VOC), such as α‐terpineol, p‐menth‐1‐en‐9‐al, linalool and 3,6‐dimethyl‐2,3,3a,4,5,7a‐hexahydrobenzofuran (DMHEX), a novel volatile recently identified in tomato. Two sensory attributes – sweetness and a not‐acidic taste – correlated with the characteristic tomato taste, and also with fructose, glucose, and two VOCs, benzaldehyde, and 2‐methyl‐2‐octen‐4‐one. CONCLUSIONS These data provide new evidence of the complex chemical combination that induced the flavour and aroma of the good‐tasting ‘criollo’ tomato fruit. That is, the compounds that correlated with defined tomato aroma and acidic taste did not correlate with sweetness, or with characteristic tomato taste.Instituto de BiotecnologíaFil: D'Angelo, Matilde. Universidad Nacional de Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Zanor, María I. Universidad Nacional de Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sance, María. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias; ArgentinaFil: Cortina, Pablo Ramiro. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Orgánica; ArgentinaFil: Boggio, Silvana B. Universidad Nacional de Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Asprelli, Pablo. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias; ArgentinaFil: Carrari, Fernando. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Universidade de São Paulo. Departamento de Botânica. Instituto de Biociências; BrasilFil: Santiago, Ana N. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Orgánica; ArgentinaFil: Asis, Ramón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas; ArgentinaFil: Peralta, Iris Edith. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mendoza. Instituto Argentino de Investigaciones de Zonas Aridas; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias; ArgentinaFil: Valle, Estela M. Universidad Nacional de Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Differential diagnosis of neurodegenerative dementias with the explainable MRI based machine learning algorithm MUQUBIA

Biomarker-based differential diagnosis of the most common forms of dementia is becoming increasingly important. Machine learning (ML) may be able to address this challenge. The aim of this study was to develop and interpret a ML algorithm capable of differentiating Alzheimer's dementia, frontotemporal dementia, dementia with Lewy bodies and cognitively normal control subjects based on sociodemographic, clinical, and magnetic resonance imaging (MRI) variables. 506 subjects from 5 databases were included. MRI images were processed with FreeSurfer, LPA, and TRACULA to obtain brain volumes and thicknesses, white matter lesions and diffusion metrics. MRI metrics were used in conjunction with clinical and demographic data to perform differential diagnosis based on a Support Vector Machine model called MUQUBIA (Multimodal Quantification of Brain whIte matter biomArkers). Age, gender, Clinical Dementia Rating (CDR) Dementia Staging Instrument, and 19 imaging features formed the best set of discriminative features. The predictive model performed with an overall Area Under the Curve of 98%, high overall precision (88%), recall (88%), and F1 scores (88%) in the test group, and good Label Ranking Average Precision score (0.95) in a subset of neuropathologically assessed patients. The results of MUQUBIA were explained by the SHapley Additive exPlanations (SHAP) method. The MUQUBIA algorithm successfully classified various dementias with good performance using cost-effective clinical and MRI information, and with independent validation, has the potential to assist physicians in their clinical diagnosis

Metabolismo de frutos con diferentes características organolépticas de especies de tomates silvestres y cultivados

El tomate cultivado (Solanum lycopersicum L.) es una especie americana domesticada para consumo humano. En Argentina los tomates cultivados son híbridos importados seleccionados por sus propiedades de larga vida postcosecha y resistencia a plagas, por lo que actualmente los consumidores no se encuentran complacidos en relación al aroma y al sabor de los frutos. En este trabajo se efectuó el análisis sensorial y la evaluación del perfil metabólico global mediante la cuantificación de 26 metabolitos solubles por 1H-RMN y de 100 compuestos volátiles por SPME- GC-MS en muestras de pericardio de frutos maduros de una colección de accesiones de tomate cultivadas en los valles de los Andes Argentinos con alta diversidad morfo-agronómica y metabólica. Mediante un análisis sensorial se identificaron las accesiones más contrastantes siendo 557, 572, y 569 las de mejor calidad mientras que 571 se evaluó como la menos sabrosa. La composición metabólica de las accesiones de calidad sobresaliente se correspondió con el contenido elevado de moléculas descriptas en la bibliografía como de impacto positivo en el sabor del tomate: glucosa, fructosa, sacarosa, citrato, Asp, Asn, Ala, 3-hexenal, 3-metil-butanal, nitropentano, 6-metil-5-hepten-2-ona, 2-isobutiltiazol, metil-salicilato, metil-butanoato, 4- pentenal, limoneno, linalool, 2-nonen-1-ol, alcanfor, β- damascenona, β-ionone epóxido, α- etillbenzyl alcohol, óxido de limoneno, fenilacetaldehído, 2-fenil-etanol, 2-undecanona, benzofenona, 8-pentadecanona, 2-heptenal, carvona y hexanal. Mediante un análisis multivariado de los datos por correlación de Pearson se identificaron 13 metabolitos solubles y 32 compuestos volátiles de impacto positivo en el aroma y el sabor, siendo el 71 % moléculas aun no citadas en la literatura como los compuestos α-citral y 2,4-decadienal. La comparación de la composición metabólica de las accesiones de calidad con la de otras no catadas permitió postular a las accesiones 565, 3834, 558, 574, 3831, 573 y 570 como de buen aroma y sabor. Además, los ACP obtenidos a partir de los datos de metabolitos primarios de los frutos de accesiones cosechadas en paralelo en dos localidades distintas indicaron que la composición metabólica de los mismos es estable en el tiempo y en el espacio. Por último, el análisis de la expresión génica global mediante microarreglos de Affymetrix en pericarpio de frutos maduros de 10 accesiones de tomate estableció que las accesiones 557 y 572 de alta calidad organoléptica presentaron perfiles transcripcionales similares entre sí y diferentes al de la accesión menos sabrosa 571. Así mismo, en dichas accesiones se identificaron los genes Solyc09g075820, Solyc05g050010, Solyc06g073730, Solyc09g075420, Solyc02g093150, Solyc01g109300 y Solyc01g006540 de expresión significativamente más altos en comparación con la accesión 571 y cuyos productos están implicados en los procesos de transporte de glucosa, biosíntesis y transducción de etileno, síntesis de monoterpenos y sesquiterpenos y catabolismo de ácidos grasos, que conjuntamente favorecen la producción y acumulación de moléculas asociadas a los parámetros de calidad organoléptica actualmente demandados. Dichos genes tienen relevancia biotecnológica ya que podrían emplearse en la construcción de plantas de tomate transgénicas de aroma y sabor mejorado.Fil: D’Angelo, Matilde Elvira. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET); Argentina

Reduced levels of nadh-dependent glutamate dehydrogenase decrease the glutamate content of ripe tomato fruit but have no effect on green fruit or leaves

Glutamate (Glu) is a taste enhancer that contributes to the characteristic flavour of foods. In fruit of tomato (Solanum lycopersicum L.), the Glu content increases dramatically during the ripening process, becoming the most abundant free amino acid when the fruit become red. There is also a concomitant increase in NADH-dependent glutamate dehydrogenase (GDH) activity during the ripening transition. This enzyme is located in the mitochondria and catalyses the reversible amination of 2-oxoglutarate to Glu. To investigate the potential effect of GDH on Glu metabolism, the abundance of GDH was altered by artificial microRNA technology. Efficient silencing of all the endogenous SlGDH genes was achieved, leading to a dramatic decrease in total GDH activity. This decrease in GDH activity did not lead to any clear morphological or metabolic phenotype in leaves or green fruit. However, red fruit on the transgenic plants showed markedly reduced levels of Glu and a large increase in aspartate, glucose and fructose content in comparison to wild-type fruit. These results suggest that GDH is involved in the synthesis of Glu in tomato fruit during the ripening processes. This contrasts with the biological role ascribed to GDH in many other tissues and species. Overall, these findings suggest that GDH has a major effect on the control of metabolic composition during tomato fruit ripening, but not at other stages of development

Reduced levels of nadh-dependent glutamate dehydrogenase decrease the glutamate content of ripe tomato fruit but have no effect on green fruit or leaves

Glutamate (Glu) is a taste enhancer that contributes to the characteristic flavour of foods. In fruit of tomato (Solanum lycopersicum L.), the Glu content increases dramatically during the ripening process, becoming the most abundant free amino acid when the fruit become red. There is also a concomitant increase in NADH-dependent glutamate dehydrogenase (GDH) activity during the ripening transition. This enzyme is located in the mitochondria and catalyses the reversible amination of 2-oxoglutarate to Glu. To investigate the potential effect of GDH on Glu metabolism, the abundance of GDH was altered by artificial microRNA technology. Efficient silencing of all the endogenous SlGDH genes was achieved, leading to a dramatic decrease in total GDH activity. This decrease in GDH activity did not lead to any clear morphological or metabolic phenotype in leaves or green fruit. However, red fruit on the transgenic plants showed markedly reduced levels of Glu and a large increase in aspartate, glucose and fructose content in comparison to wild-type fruit. These results suggest that GDH is involved in the synthesis of Glu in tomato fruit during the ripening processes. This contrasts with the biological role ascribed to GDH in many other tissues and species. Overall, these findings suggest that GDH has a major effect on the control of metabolic composition during tomato fruit ripening, but not at other stages of development

An insight into the commercial piglet’s microbial gut colonization: from birth towards weaning

Abstract Background The establishment of the gut microbiota can be influenced by several perinatal factors, including, most importantly, the maternal microbiota. Moreover, early-life environmental variation affects gut microbial colonization and the intestinal health of offspring throughout life. The present study aimed to explore the development of piglet gut microbiota from birth to weaning in the commercial practice and also to assess how different farm environments could condition this process. Although it is possible to find in the literature other studies with similar objectives this work probably represents one of the few studies that make a systematic evaluation of such differential factors under a real scenario. To achieve this objective, we performed two trials. In a first Trial, we selected 2 farms in which we performed an intensive sampling (5 samples /animal) to characterize the gut colonization pattern during the first days of life and to identify the time window with the greatest impact. Both farms differed in their health status and the use of antimicrobials in the piglets. In a second Trial, we selected 4 additional farms with variable rearing conditions and a distinctive use of antimicrobials in the sows with a simplified sampling pattern (2 samples/animal). Faecal samples were obtained with swabs and DNA was extracted by using the PSP® Spin Stool DNA Kit and sequencing of the 16S rRNA gene (V3-V4 region) performed by Illumina MiSeq Platform. Results The present study contributes to a better understanding of microbiome development during the transition from birth to weaning in commercial conditions. Alpha diversity was strongly affected by age, with an increased richness of species through time. Beta diversity decreased after weaning, suggesting a convergent evolvement among individuals. We pinpointed the early intestinal colonizers belonging to Bacteroides, Escherichia-Shigella, Clostridium sensu stricto 1, and Fusobacterium genera. During lactation(d7-d21 of life), the higher relative abundances of Bacteroides and Lactobacillus genera were correlated with a milk-oriented microbiome. As the piglets aged and after weaning (d36 of life), increasing abundances of genera such as Prevotella, Butyricimonas, Christensenellaceae R-7 group, Dorea, Phascolarctobacterium, Rikenellaceae RC9 gut group, Subdoligranulum, and Ruminococcaceae UCG-002 were observed. These changes indicate the adaptation of the piglets to a cereal-based diet rich in oligosaccharides and starch. Our results also show that the farm can have a significant impact in such a process, evidencing the influence of different environments and rearing systems on the gut microbiota development of the young piglet. Differences between farms were more noticeable after weaning than during lactation with changes in alpha and beta biodiversity and specific taxa. The analysis of such differences suggests that piglets receiving intramuscular amoxicillin (days 2–5 of life) and being offered an acidifying rehydrating solution (Alpha farm in Trial 1) have a greater alpha diversity and more abundant Lactobacillus population. Moreover, the only farm that did not offer any rehydrating solution (Foxtrot farm in Trial 2) showed a lower alpha diversity (day 2 of life) and increased abundance of Enterobacteriaceae (both at 2 and 21 days). The use of in-feed antibiotics in the sows was also associated with structural changes in the piglets’ gut ecosystem although without changes in richness or diversity. Significant shifts could be registered in different microbial groups, particularly lower abundances of Fusobacterium in those piglets from medicated sows. Conclusions In conclusion, during the first weeks of life, the pig microbiota showed a relevant succession of microbial groups towards a more homogeneous and stable ecosystem better adapted to the solid dry feed. In this relevant early-age process, the rearing conditions, the farm environment, and particularly the antimicrobial use in piglets and mothers determine changes that could have a relevant impact on gut microbiota maturation. More research is needed to elucidate the relative impact of these farm-induced early life-long changes in the growing pig

Fruit metabolic and transcriptional programs differentiate among Andean tomato (Solanum lycopersicum L.) accessions

Traditional landraces or “criollo” tomatoes (Solanum lycopersicum L.) from Andean areas of Argentina, selected for their fruit quality, were analysed in this study. We explored the metabolome and transcriptome of the ripe fruit in nine landrace accessions representing the seven genetic groups and compared them to the mature fruit of the wild progenitor Solanum pimpinellifolium. The content of branched- (isoleucine and valine) and aromatic (phenylalanine and tryptophan) amino acids, citrate and sugars were significantly different in the fruit of several “criollo” tomatoes compared to S. pimpinellifolium. The transcriptomic profile of the ripe fruit showed several genes significantly and highly regulated in all varieties compared to S. pimpinellifolium, like genes encoding histones and mitochondrial proteins. Additionally, network analysis including transcripts and metabolites identified major hubs with the largest number of connections such as constitutive photomorphogenic protein 1 (a RING finger-type ubiquitin E3 ligase), five Zn finger transcription factors, ascorbate peroxidase, acetolactate synthase, and sucrose non-fermenting 1 kinase. Co-expression analysis of these genes revealed a potential function in acquiring tomato fruit quality during domestication.EEA La ConsultaFil: D'Angelo, Matilde. Universidad Nacional de Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universitat Autónoma de Barcelona. Animal and Food Science Department. Animal Nutrition and Welfare Service; EspañaFil: Zanor, María Inés. Universidad Nacional de Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Burgos, Estanislao. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina.Fil: Asprelli, Pablo. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria La Consulta; ArgentinaFil: Boggio, Silvana Beatriz. Universidad Nacional de Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Carrari, Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina.Fil: Peralta, Iris Edith. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mendoza. Instituto Argentino de Investigaciones de Zonas Aridas; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias; ArgentinaFil: Valle, Estela M. Universidad Nacional de Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Role of rMnSOD as Cytotoxic Agent and Chemotherapy Enhancer in Paediatric Acute Lymphoblastic Leukemia B Cells

Background/Objectives: An isoform of Manganese-superoxide-dismutase (rMnSOD) was isolated and sequenced for the first time from a human Liposarcoma cells and obtained in recombinant form. The protein showed a specific and selective cytotoxicity on many cancer cell lines. rMnSOD enters cells by means of its 24-aa leader peptide that linked to the oestrogen receptor (ER). A 6-aa sequence, that participates in ER binding, was identified as a molecular carrier by mass spectrometric analysis. The aim of this study was to test the ability of this molecule to inhibit the growth of SUP-B15 cell line and leukemic B-cells and kill them alone and as enhancer of chemotherapic drug. Design/Methods: SUP-B15 cell line and leukemic B cells were cultured in RPMI medium and treated for 5 hours with rMnSOD and Daunoblastine, single and in combination. Cell viability assay was analyzed by MUSE analyzer (Millipore). Apoptotic cell death and cell cycle were analyzed by Annexin-V-FITC staining and PI fluorescence.We analyzed ER expression levels by Real time PCR. Results: A high level of ER was observed in SUP-B15. Our preliminary data showed that in SUP-B15 cell line rMnSOD induced apoptosis with a reduction of vitality from 61.9% to 31.40% respect to untreated. Treatment with rMnSOD of leukemic cell samples showed in all apoptotic profiles an increase in late apoptosis up to 52%. Better apoptotic induction was observed in prednisone poor responders. Cell cycle analysis showed an arrest in Go/G1 after treatment. Conclusion: rMnSOD is a promising molecule for the treatment of paediatric B-ALL due to its selective action to cancer cells and to its enhancer action which leads to reduction of drug dose and significantly reducing side effects. Targeted delivery can increase the therapeutic index of conventional chemotherapy, with low doses to kill cancer cells

Microenvironment in neuroblastoma: isolation and characterization of tumor-derived mesenchymal stromal cells

Abstract Background It has been proposed that mesenchymal stromal cells (MSCs) promote tumor progression by interacting with tumor cells and other stroma cells in the complex network of the tumor microenvironment. We characterized MSCs isolated and expanded from tumor tissues of pediatric patients diagnosed with neuroblastomas (NB-MSCs) to define interactions with the tumor microenvironment. Methods Specimens were obtained from 7 pediatric patients diagnosed with neuroblastoma (NB). Morphology, immunophenotype, differentiation capacity, proliferative growth, expression of stemness and neural differentiation markers were evaluated. Moreover, the ability of cells to modulate the immune response, i.e. inhibition of phytohemagglutinin (PHA) activated peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cytotoxic function, was examined. Gene expression profiles, known to be related to tumor cell stemness, Wnt pathway activation, epithelial-mesenchymal transition (EMT) and tumor metastasis were also evaluated. Healthy donor bone marrow-derived MSCs (BM-MSC) were employed as controls. Results NB-MSCs presented the typical MSC morphology and phenotype. They showed a proliferative capacity superimposable to BM-MSCs. Stemness marker expression (Sox2, Nanog, Oct3/4) was comparable to BM-MSCs. NB-MSC in vitro osteogenic and chondrogenic differentiation was similar to BM-MSCs, but NB-MSCs lacked adipogenic differentiation capacity. NB-MSCs reached senescence phases at a median passage of P7 (range, P5-P13). NB-MSCs exhibited greater immunosuppressive capacity on activated T lymphocytes at a 1:2 (MSC: PBMC) ratio compared with BM-MSCs (p = 0.018). NK cytotoxic activity was not influenced by co-culture, either with BM-MSCs or NB-MSCs. Flow-cytometry cell cycle analysis showed that NB-MSCs had an increased number of cells in the G0-G1 phase compared to BM-MSCs. Transcriptomic profiling results indicated that NB-MSCs were enriched with EMT genes compared to BM-MSCs. Conclusions We characterized the biological features, the immunomodulatory capacity and the gene expression profile of NB-MSCs. The NB-MSC gene expression profile and their functional properties suggest a potential role in promoting tumor escape, invasiveness and metastatic traits of NB cancer cells. A better understanding of the complex mechanisms underlying the interactions between NB cells and NB-derived MSCs should shed new light on potential novel therapeutic approaches