Maturation curves of sweet sorghum genotypes.

O sorgo sacarino [Sorghum bicolor (L.) Moench] se destaca como cultura complementar à cana-de-açúcar Saccharum spp. na produção de etanol, uma vez que este possui colmos suculentos com presença de açúcares diretamente fermentescíveis. Em decorrência deste fato, existe a necessidade de se analisar as propriedades do sorgo, a fim de atender às demandas da agroindústria. No presente trabalho, buscou-se desenvolver e estudar as curvas de maturação de sete cultivares de sorgo sacarino em dez épocas de colheita. Os resultados mostram que houve diferença significativa para cultivares e épocas de colheita para todos os parâmetros (p≤0,01). Quanto aos teores de açúcares, as cultivares BRS508, XBWS80147 e CMSXS629 apresentaram as maiores médias para as variáveis açúcares redutores totais (ART) e açúcares totais recuperáveis (ATR). Em relação à produção de etanol por tonelada de biomassa, as cultivares BRS 508 e a CMSXS629 apresentaram os melhores resultados.

Régime thermique en période de maturation du raisin dans le géoclimat viticole.

Mots clés :indice de fraîcheur des nuits, amplitude thermique, Système CCM Géoviticole, zonage, qualité. Key words: cool night index, thermal amplitude, MCC System for World Viticulture, zoning, quality. RESUME Le régime thermique en période de maturation du raisin est l?une des variables déterminantes de la coloration du raisin et de la richesse en arômes, anthocyanes et polyphénols des vins. L?objectif du travail est de caractériser le régime thermique, notamment la fraîcheur des nuits et l?amplitude thermique au cours de la maturation, au niveau du climat viticole mondial, sur une base des données de 100 régions viticoles dans 30 pays, obtenue par l?intermédiaire de l?Organisation Mondiale de la Météorologie - OMM. Plusieurs indices climatiques viticoles ont été calculés: l?Indice de Fraîcheur des Nuits ?IF (ºC), l?Indice Héliothermique de Huglin ?IH (ºC) et l?Indice de Sécheresse ?IS (mm) du Système de Classification Climatique Multicritères Géoviticole, et l?amplitude thermique moyenne en août et septembre Aa-s (ºC). Egalement, sur la période véraison-récolte ?v-r(moyenne des 30 jours précédant la date de récolte, estimée sur la base d?un Indice Héliothermique de HUGLIN égal à 1.900 - approximatif pour la maturation du Cabernet-Sauvignon): la fraîcheur des nuits (FNv -r), la température moyenne de l?air (Tv-r), la température maximale de l?air (Txv-r) et l?amplitude thermique (Av-r). Les résultats montrent que IH est corrélé avec Tv-r (r=0,79**) et avec Txv-r (r=0,80**). IH représente donc bien les conditions thermiques générales de la période de maturation en ce qui concerne la température moyenne et maximale de l?air. Mais IH n?est pas corrélé ni avec Aa-s ni avec Av-r. Par contre, IF est corrélé avec Aa-s (r=-0,70**) et FNv-r est corrélé avec Av-r (r=-0,69**). Cette corrélation doit justifier, en partie, l?usage assez courant de l?amplitude thermique comme indicateur de bonnes conditions thermiques de maturation pour les régions qui présentent des valeurs élevées. Mais ce raisonnement peut amener à des caractérisations erronées. Le travail met en évidence, également, ?importance de considérer le bilan hydrique des régions (IS) dans l?analyse du régime thermique sur la qualité du raisin. On peut onclure que pour avoir une bonne caractérisation du régime thermique en période de maturation il faut considérer la fraîcheur des nuits (IF étant un bon indicateur de FNv-r moyen des régions, avec un r=0,80**), caractérisation qui peut être améliorée avec l?information des températures maximales et de sur 11l?amplitude thermique en période de maturation du raisin. Les éléments présentés peuvent servir à améliorer les indices climatiques pour estimer le potentiel qualitatif du raisin des différentes régions viticoles, notamment en complément de IF. ABSTRACT The thermal conditions during the grape ripening period are important variables related to colour of the grapes, anthocyanins, polyphenols and flavour of the wine. The main purpose of this work was to characterise the thermal conditions, especially the night coolness and the thermal amplitude during maturation, in the geoclimate of the world vine culture. A database of 100 grape-growing regions of 30 countries obtained from the World Meteorology Organisation (WMO) was used. Some climatic indexes were calculated: Cool Night Index ?IF (°C), Huglin?s Heliothermal Index ?IH (°C) and Dryness Index ?IS (mm), from the Multicriteria Climatic Classification System for World Viticulture, and the thermal amplitude in August and September Aa-s (°C). Over véraison-harvest period ?v-r(mean of the 30 days before harvesting date, estimated on the basis of HUGLIN Heliothermal Index equal to 1,900 ? approximately value to ripen Cabernet-Sauvignon) similar indexes were obtained: the cool night (FNv-r), the mean air temperature (Tv-r), the maximal air temperature (Txv-r) and the thermal amplitude (Av-r). The results showed that IH is positively correlated with Tv-r (r=0.79**), Txv-r (r=0.80)and IF (r=0.67**). Therefore, IH represents well the general thermal conditions during maturation period, specially concerning the mean and the maximal air temperature. owever, IH was correlated neither with Aa-s nor to Av-r. IF was negatively correlated with Aa-s (r= -0.70**) and FNv-r was negatively correlated with Av-r (r=-0.69**). The correlation to some extent explains the current use of the thermal amplitude to predict good ripening thermal conditions for those regions that show high values. As here we have described, this thinking may give incorrect results. This work has also showed the importance to consider the water balance of the regions (IS) in the effect of the thermal conditions in grape quality. We conclude that the characterisation of the thermal conditions during the ripening period do need the cool night index (in this case, IFis a good index to provide the mean FNv-r of the regions, r=0,80**). Factors other than cool night which influence this characterisation are both maximal air temperature and thermal amplitude data. The elements presented in this work, in addition to IF , may improve the climatic indexes to be used to predict the qualitative potential of grapes from different regions.

Adaptability and phenotypic stability of soybean cultivars for grain yield and oil content.

ABSTRACT. The aim of this study was to verify the adaptability and stability of soybean cultivars with regards to yield and oil content. Data of soybean yield and oil content were used from experiments set up in six environments in the 2011/12 and 2012/13 crop seasons in the municipalities of Patos de Minas, Uberaba, Lavras, and São Gotardo, Minas Gerais, Brazil, testing 36 commercial soybean cultivars of both conventional and transgenic varieties. The Wricke method and GGE biplot analysis were used to evaluate adaptability and stability of these cultivars. Large variations were observed in grain yield in relation to the different environments studied, showing that these materials are adaptable. The cultivars exhibited significant differences in oil content. The cultivars BRSGO204 (Goiânia) and BRSMG (Garantia) exhibited the greatest average grain yield in the different environments studied, and the cultivar BRSMG 760 SRR had the greatest oil content among the cultivars evaluated. Ecovalence was adopted to identify the most stable cultivars, and the estimates were nearly uniform both for grain yield and oil content, showing a variation of 0.07 and 0.01%, respectively. The GGE biplot was efficient at identifying cultivars with high adaptability and phenotype stability.

Exogenous paraoxonase-1 during oocyte maturation improves bovine embryo development in vitro.

The aims of this study were (i) to measure the direct effects of exogenous human recombinant PON1 (rPON1) on bovine oocyte maturation at the molecular level (gene expression) and (ii) to measure the carry-over effects of PON1 on pre-implantation embryo development in vitro.

Maturation behavior of Maxi Gala grafetd on two rootstocks by no Destructive Method.

The evaluation of the maturation in apple orchards is checked using destructive methods, sampling fruits and analyzing them in the laboratory, making the process slow and expensive. The use of not destructive method to determine fruit maturation in the orchard could accelerate delivery of results and help in determining harvest time, because non-destructive data would allow to verify the maturation on different blocks in the orchard. The aim of this work was to chart fruit maturation in 'Maxi Gala' grafted on two different rootstocks, using destructive and not destructive methods. The non-destructive method used was the portable DA-Meter. The trial was realized at Vacaria, southern Brazillocated 28,44 S and 50,85 W. The samples were harvested on two orchards during the seasons 2014/15 and 2015/16, during six weeks before harvest from January until the second week of February. The sampling was realized in five different points of the orchard, on rootstocks M.9 or Marubakaido with M.9 interstem. Ten-apple samples were collected weekly in each point in the orchard and then evaluated by destructive method (flesh firmness, starch degradation, total soluble solids and acidity) and the not destructive method (DA-Meter). For both seasons, the evolution of the fruit maturation of Maxi Gala showed a similar progression for both rootstocks. The non-destructive method correlated well with the traditional destructive methods, making it a tool for more practical and easy determination of the harvest date.

Glutathione and abscisic acid supplementation influences somatic embryo maturation and hormone endogenous levels during somatic embryogenesis in Podocarpus lambertii Klotzsch ex Endl.

Here we propose a protocol for embryogenic cultures induction, proliferation and maturation for the Brazilian conifer Podocarpus lambertii, and investigated the effect of abscisic acid (ABA) and glutathione (GSH) supplementation on the maturation phase. ABA, zeatin (Z) and salicylic acid (SA) endogenous levels were quantified. Number of somatic embryos obtained in ABA-supplemented treatment was signifi- cant higher than in ABA-free treatment, showing the relevance of ABA supplementation during somatic embryos maturation. Histological analysis showed the stereotyped sequence of developmental stages in conifer somatic embryos, reaching the late torpedo-staged embryo. GSH supplementation in maturation culture medium improved the somatic embryos number and morphological features. GSH 0 mM and GSH 0.1 mM treatments correlated with a decreased ABA endogenous level during maturation, while GSH 0.5 mM treatment showed constantlevels. Alltreatments resulted in decreased Z endogenous levels, supporting the concept that cytokinins are important during the initial cell division but not for the later stages of embryo development. The lowest SA levels found in GSH 0.5 mM treatment were coincident with early embryonic development, and this treatment resulted in the highest development of somatic embryos. Thus, a correlation between lower SA levels and improved somatic embryo formation can be hypothesized

Wavelet analysis and hierarchical clustering of ofm phenotypic population's variations originated from southern america and europe and attempts of a first relation to molecular divergence.

In this work we compare Grapholita molesta Busck (Lepidoptera: Tortricidae) populations originated from Brazil, Chile, Spain, Italy and Greece using power spectral density and phylogenetic analysis to detect any similarities between the population macro- and the molecular micro-level. Log-transformed population data were normalized and AR(p) models were developed to generate for each case population time series of equal lengths. The time-frequency/scale properties of the population data were further analyzed using wavelet analysis to detect any population dynamics frequency changes and cluster the populations. Based on the power spectral of each population time series and the hierarchical clustering schemes, populations originated from Southern America (Brazil and Chile) exhibit similar rhythmic properties and are both closer related with populations originated from Greece. Populations from Spain and especially Italy, have higher distance by terms of periodic changes on their population dynamics. Moreover, the members within the same cluster share similar spectral information, therefore they are supposed to participate in the same temporally regulated population process. On the contrary, the phylogenetic approach revealed a less structured pattern that bears indications of panmixia, as the two clusters contain individuals from both Europe and South America. This preliminary outcome will be further assessed by incorporating more individuals and likely employed a second molecular marker.

Effect of host genotype and prion strain on the phenotypic heterogeneity of Creutzfeldt-Jakob disease: the peripheral nervous system involvement and the spectrum of the genetic form

The first study was designed to assess whether the involvement of the peripheral nervous system (PNS) belongs to the phenotypic spectrum of sporadic Creutzfeldt-Jakob disease (sCJD). To this aim, we reviewed medical records of 117 sCJDVV2, 65 sCJDMV2K, and 121 sCJDMM(V)1 subjects for symptoms/signs and neurophysiological data. We looked for the presence of PrPSc in postmortem PNS samples from 14 subjects by western blotting and real-time quaking-induced conversion (RT-QuIC) assay. Seventy-five (41.2%) VV2-MV2K patients, but only 11 (9.1%) MM(V)1, had symptoms/signs suggestive of PNS involvement and neuropathy was documented in half of the VV2-MV2K patients tested. RT-QuIC was positive in all PNS samples, whereas western blotting detected PrPSc in the sciatic nerve in only one VV2 and one MV2K. These results support the conclusion that peripheral neuropathy, likely related to PrPSc deposition, belongs to the phenotypic spectrum of sCJDMV2K and VV2, the two variants linked to the V2 strain. The second study aimed to characterize the genetic/molecular determinants of phenotypic variability in genetic CJD (gCJD). To this purpose, we compared 157 cases of gCJD to 300 of sCJD. We analyzed: demographic aspects, neurological symptoms/signs, histopathologic features and biochemical characteristics of PrPSc. The results strongly indicated that the clinicopathological phenotypes of gCJD largely overlap with those of sCJD and that the genotype at codon 129 in cis with the mutation (i.e. haplotype) contributes more than the latter to the disease phenotype. Some mutations, however, cause phenotypic variations including haplotype-specific patterns of PrPSc deposition such as the “dense” synaptic pattern (E200K-129M), the intraneuronal dots (E200K-129V), and the linear stripes perpendicular to the surface in the molecular layer of cerebellum (OPRIs-129M). Overall, these results suggest that in gCJD PRNP mutations do not cause the emergence of novel prion strains, but rather confer increased susceptibility to the disease in conjunction with “minor” clinicopathological variations.

Phenotypic and genotypic characterisation of antimicrobial resistance in Escherichia coli indicator of animal, food and human origin.

This thesis presents AMR phenotypic evaluation and whole genome sequencing analysis of 288 Escherichia coli strains isolated from different sources (livestock, companion animal, wildlife, food and human) in Italy. Our data reflects general resistance trends in Europe, reporting tetracycline, ampicillin, sulfisoxazole and aminoglycosides resistance as the most common phenotypic AMR profile among livestock, pets, wildlife and humans. Identification of human and animal (livestock and companion animal) AMR profiles in niches with a rare (fishery, mollusc) or absent (vegetable, wild animal, wild boar) direct exposure to antimicrobials, suggests widespread environmental pollution with ARGs conferring resistance to these antimicrobials. Phenotypic resistance to highest priority critically important antimicrobials was mainly observed in food-producing animals and related food such as rabbit, poultry, beef and swine. Discrepancies between AMR phenotypic pattern and genetic profile were observed. In particular, phenotypic aminoglycoside, cephalosporin, meropenem, colistin resistance and ESBL profile did not have a genetic explanation in different cases. This data could suggest the diffusion of new genetic variants of ARGs, associated to these antimicrobial classes. Generally, our collection shows a virulence profile typical of extraintestinal pathogenic Escherichia coli (ExPEC) pathotype. Different pandemic and emerging ExPEC lineages were identified, in particular in poultry meat (ST10; ST23; ST69, ST117; ST131). Rabbit was suggested as a source of ST20-ST40 potential hybrid pathogens. Wildlife carried a high average number (10) of VAGs (mostly associated to ExPEC pathotype) and different predominant ExPEC lineages (ST23, ST117, ST648), suggesting its possible involvement in maintenance and diffusion of virulence determinants. In conclusion, our study provides important knowledge related to the phenotypic/genetic AMR and virulence profiles circulating in E. coli in Italy. The role of different niches in AMR dynamics has been discussed. In particular, food-producing animals are worthy of continued investigation as a source of potential zoonotic pathogens, meanwhile wildlife might contribute to VAGs spread.

Feeding the Gut Microbiome and Immune Maturation to Manage Weaned Piglets

This thesis reports five studies that may contribute to understand how weaning affects the immune and intestinal microbiota maturation of the piglet and proposes some possible nutritional strategies to attenuate its negative effects. The first study showed that weaning is associated in Payer’s patches with the activation of MHC response against class I antigens and that related to the stimulation to IFN-γ and showed, for the first time, that their blood at weaning remains dominated by immature blood cells. In the second study we tested if the use of a live vaccine against a conditionally but also genetically based intestinal disease, like PWD, could have an impact on the growth performance of pigs and their intestinal microbiota and if it could provide a model to test the response to nutritional strategies under conditions of an immune and intestinal stimulation for animals susceptible to ETEC type. In this study, we demonstrated how a vaccinal strain of F4/F18 E. coli can affect the gut microbial composition of piglets, regardless of their genetic susceptibility to ETEC infection. In the third study we evidenced how a nucleotide supplementation can favor the proliferation of jejunal Peyer patches and anticipate the maturation of the fecal microbiota. In the fourth study we reported how xylanase can favor the proliferation of Lactobacillus reuteri. Finally, we showed some first results on the muscles fiber development in fast- and slow-growing suckling pigs and the relationship with the intestinal microbiota. Taken together, the results presented in this thesis provide new insight about the interplay between the host-genetics, gut microbial composition, and host physiological status. Furthermore, it provides confirmation that the use of known genetic markers for ETEC F4 and F18 could represent a potential tool to stratify the animals in the trials both in healthy or challenge-based protocols.

Stayability in Italian Simmental dual-purpose cows: phenotypic and genetic associations

In recent years, dairy farmers have observed a substantial decrease in cows’ survival, with a direct negative consequence on the profitability. Shorter lifespan raises questions about animal welfare and farming conditions at which cows are exposed to. However, the length of productive life depends also on voluntary culling due to low productivity and, in dual-purpose breed, to low price of carcasses (meat). The general aim of the thesis was to investigate the genetic and phenotypic relationship of functional longevity with morphological features like muscularity and body condition score (BCS) and productive traits within Italian Simmental dual-purpose dairy cattle raised in Emilia-Romagna herds.

Phenotypic and molecular investigation of circulating tumor cells (CTCs) isolated from breast cancer patients at single cell resolution

Despite the paramount advances in cancer research, breast cancer (BC) still ranks one of the leading causes of cancer-related death worldwide. Thanks to the screening campaign started in developed countries, BC is often diagnosed at early stages (non-metastatic BC, nmBC), but disease relapse occurrence even after decades and at distant sites is not an uncommon phenomenon. Conversely, metastatic BC (mBC) is considered an incurable disease. The major perpetrators of tumor spread to secondary organs are circulating tumor cells (CTCs), a rare population of cells detectable in the peripheral blood of oncologic patients. In this study, CTCs from patients diagnosed with luminal nmBC and mBC (hormone receptor positive, Human Epidermal Growth Factor Receptor 2 (HER2) negative) were characterized at both phenotypic and molecular levels. To better understand the molecular mechanisms underlying their biology and their metastatic potential, next-generation sequencing (NGS) analyses were performed at single-cell resolution to assess copy number aberrations (CNAs), single nucleotide variants (SNVs) and gene expression profiling. The findings of this study arise hints in CTC detection, and pave the way to new application in CTC research.

An Efficient Protocol For The Generation Of Monocyte Derived Dendritic Cells Using Serum-free Media For Clinical Applications In Post Remission Aml Patients.

Protocols for the generation of dendritic cells (DCs) using serum as a supplementation of culture media leads to reactions due to animal proteins and disease transmissions. Several types of serum-free media (SFM), based on good manufacture practices (GMP), have recently been used and seem to be a viable option. The aim of this study was to evaluate the results of the differentiation, maturation, and function of DCs from Acute Myeloid Leukemia patients (AML), generated in SFM and medium supplemented with autologous serum (AS). DCs were analyzed by phenotype characteristics, viability, and functionality. The results showed the possibility of generating viable DCs in all the conditions tested. In patients, the X-VIVO 15 medium was more efficient than the other media tested in the generation of DCs producing IL-12p70 (p=0.05). Moreover, the presence of AS led to a significant increase of IL-10 by DCs as compared with CellGro (p=0.05) and X-Vivo15 (p=0.05) media, both in patients and donors. We concluded that SFM was efficient in the production of DCs for immunotherapy in AML patients. However, the use of AS appears to interfere with the functional capacity of the generated DCs.

Molecular And Biochemical Characterization Of Caffeine Synthase And Purine Alkaloid Concentration In Guarana Fruit.

Guarana seeds have the highest caffeine concentration among plants accumulating purine alkaloids, but in contrast with coffee and tea, practically nothing is known about caffeine metabolism in this Amazonian plant. In this study, the levels of purine alkaloids in tissues of five guarana cultivars were determined. Theobromine was the main alkaloid that accumulated in leaves, stems, inflorescences and pericarps of fruit, while caffeine accumulated in the seeds and reached levels from 3.3% to 5.8%. In all tissues analysed, the alkaloid concentration, whether theobromine or caffeine, was higher in young/immature tissues, then decreasing with plant development/maturation. Caffeine synthase activity was highest in seeds of immature fruit. A nucleotide sequence (PcCS) was assembled with sequences retrieved from the EST database REALGENE using sequences of caffeine synthase from coffee and tea, whose expression was also highest in seeds from immature fruit. The PcCS has 1083bp and the protein sequence has greater similarity and identity with the caffeine synthase from cocoa (BTS1) and tea (TCS1). A recombinant PcCS allowed functional characterization of the enzyme as a bifunctional CS, able to catalyse the methylation of 7-methylxanthine to theobromine (3,7-dimethylxanthine), and theobromine to caffeine (1,3,7-trimethylxanthine), respectively. Among several substrates tested, PcCS showed higher affinity for theobromine, differing from all other caffeine synthases described so far, which have higher affinity for paraxanthine. When compared to previous knowledge on the protein structure of coffee caffeine synthase, the unique substrate affinity of PcCS is probably explained by the amino acid residues found in the active site of the predicted protein.

Growth And Body Composition In Brazilian Female Rhythmic Gymnastics Athletes.

The aim was to analyse the physical growth and body composition of rhythmic gymnastics athletes relative to their level of somatic maturation. This was a cross-sectional study of 136 athletes on 23 teams from Brazil. Mass, standing height and sitting height were measured. Fat-free and fat masses, body fat percentages and ages of the predicted peak height velocity (PHV) were calculated. The z scores for mass were negative during all ages according to both WHO and Brazilian references, and that for standing height were also negative for all ages according to WHO reference but only until 12 years old according to Brazilian reference. The mean age of the predicted PHV was 12.1 years. The mean mass, standing and sitting heights, body fat percentage, fat-free mass and fat mass increased significantly until 4 to 5 years after the age of the PHV. Menarche was reached in only 26% of these athletes and mean age was 13.2 years. The mass was below the national reference standards, and the standing height was below only for the international reference, but they also had late recovery of mass and standing height during puberty. In conclusion, these athletes had a potential to gain mass and standing height several years after PHV, indicating late maturation.