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Background: In alfalfa (Medicago sativa), the coexistence of interfertile subspecies (i.e. sativa, falcata and coerulea) characterized by different ploidy levels (diploidy and tetraploidy) and the occurrence of meiotic mutants capable of producing unreduced (2n) gametes, have been efficiently combined for the establishment of new polyploids. The wealth of agronomic data concerning forage quality and yield provides a thorough insight into the practical benefits of polyploidization. However, many of the underlying molecular mechanisms regarding gene expression and regulation remained completely unexplored. In this study, we aimed to address this gap by examining the transcriptome profiles of leaves and reproductive tissues, corresponding to anthers and pistils, sampled at different time points from diploid and tetraploid Medicago sativa individuals belonging to progenies produced by bilateral sexual polyploidization (dBSP and tBSP, respectively) and tetraploid individuals stemmed from unilateral sexual polyploidization (tUSP)., Results: Considering the crucial role played by anthers and pistils in the reduced and unreduced gametes formation, we firstly analyzed the transcriptional profiles of the reproductive tissues at different stages, regardless of the ploidy level and the origin of the samples. By using and combining three different analytical methodologies, namely weighted-gene co-expression network analysis (WGCNA), tau (τ) analysis, and differentially expressed genes (DEGs) analysis, we identified a robust set of genes and transcription factors potentially involved in both male sporogenesis and gametogenesis processes, particularly in crossing-over, callose synthesis, and exine formation. Subsequently, we assessed at the same floral stage, the differences attributable to the ploidy level (tBSP vs. dBSP) or the origin (tBSP vs. tUSP) of the samples, leading to the identification of ploidy and parent-specific genes. In this way, we identified, for example, genes that are specifically upregulated and downregulated in flower buds in the comparison between tBSP and dBSP, which could explain the reduced fertility of the former compared to the latter materials., Conclusions: While this study primarily functions as an extensive investigation at the transcriptomic level, the data provided could represent not only a valuable original asset for the scientific community but also a fully exploitable genomic resource for functional analyses in alfalfa., (© 2024. The Author(s).)

A bacterial metabarcoding approach was used to compare the microbiome composition of caecal and faecal samples from fattening Japanese quails (Coturnix coturnix japonica) fed three different diet regimes. The tested feedstuffs included (1) a commercial diet for fattening quails, (2) a commercial diet containing 12% full-fat silkworm (Bombyx mori) pupae meal, and (3) a commercial diet containing 12% defatted silkworm pupae meal. The aim of the experiment was to verify the relative effect of three variables (diet type, gut tract comparing caecum to rectum, and individual animal) in determining the level of bacterial community dissimilarity to rank the relevance of each of the three factors in affecting and shaping community composition. To infer such ranking, the communities resulting from the high-throughput sequencing from each sample were used to calculate the Bray-Curtis distances in all the pairwise combinations, whereby identical communities would score 0 and totally different ones would yield the maximum distance, equal to 1. The results indicated that the main driver of divergence was the gut tract, as distances between caecal and faecal samples were higher on average, irrespective of diet composition, which scored second in rank, and of whether they had been sampled from the same individual, which was the least effective factor. Simpson's species diversity indexes was not significantly different when comparing tracts or diets, while community evenness was reduced in full-fat silkworm diet-fed animals. The identities of the differentially displayed taxa that were statistically significant as a function of gut tract and diet regimen are discussed in light of their known physiological and functional traits., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Bertoldo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Standard high-flow nasal cannula (HFNC) is a respiratory support device widely used to manage post-extubation hypoxemic acute respiratory failure (hARF) due to greater comfort, oxygenation, alveolar recruitment, humidification, and reduction of dead space, as compared to conventional oxygen therapy. On the contrary, the effects of the new asymmetrical HFNC interface (Optiflow® Duet system (Fisher & Paykel, Healthcare, Auckland, New Zealand) is still under discussion. Our aim is investigating whether the use of asymmetrical HFNC interface presents any relevant difference, compared with the standard configuration, on lung aeration (as assessed by end-expiratory lung impedance (EELI) measured by electrical impedance tomography (EIT)), diaphragm ultrasound thickening fraction (TFdi) and excursion (DE), ventilatory efficiency (estimated by corrected minute ventilation (MV)), gas exchange, dyspnea, and comfort., Methods: Pilot physiological crossover randomized controlled study enrolling 20 adults admitted to the Intensive Care unit, invasively ventilated for at least 24 h, and developing post-extubation hARF, i.e., PaO 2 /set FiO 2  < 300 mmHg during Venturi mask (VM) within 120 min after extubation. Each HFNC configuration was applied in a randomized 60 min sequence at a flow rate of 60 L/min., Results: Global EELI, TFdi, DE, ventilatory efficiency, gas exchange and dyspnea were not significantly different, while comfort was greater during asymmetrical HFNC support, as compared to standard interface (10 [7-10] and 8 [7-9], p-value 0.044)., Conclusions: In post-extubation hARF, the use of the asymmetrical HFNC, as compared to standard HFNC interface, slightly improved patient comfort without affecting lung aeration, diaphragm activity, ventilatory efficiency, dyspnea and gas exchange., Clinical Trial Number: ClinicalTrial.gov., Registration Number: NCT05838326 (01/05/2023)., New & Noteworthy: The asymmetrical high-flow nasal cannula oxygen therapy (Optiflow® Duet system (Fisher & Paykel, Healthcare, Auckland, New Zealand) provides greater comfort as compared to standard interface; while their performance in term of lung aeration, diaphragm activity, ventilatory efficiency, dyspnea, and gas exchange is similar., (© 2024. The Author(s).)

Lignohumates are increasing in popularity in agriculture, but their chemistry and effects on plants vary based on the source and processing. The present study evaluated the ability of two humates (H1 and H2) to boost maize plant performance under different phosphorus (P) availability (25 and 250 μM) conditions in hydroponics, while understanding the underlying mechanisms. Humates differed in chemical composition, as revealed via elemental analysis, phenol and phytohormone content, and thermal and spectroscopic analyses. H1 outperformed H2 in triggering plant responses to low phosphorus by enhancing phosphatase and phytase enzymes, P acquisition efficiency, and biomass production. It contained higher levels of endogenous auxins, cytokinins, and abscisic acid, likely acting together to stimulate plant growth. H1 also improved the plant antioxidant capacity, thus potentially increasing plant resilience to external stresses. Both humates increased the nitrogen (N) content and acted as biostimulants for P and N acquisition. Consistent with the physiological and biochemical data, H1 upregulated genes involved in growth, hormone signaling and defense in all plants, and in P recycling particularly under low-P conditions. In conclusion, H1 showed promising potential for effective plant growth and nutrient utilization, especially in low-P plants, involving hormonal modulation, antioxidant enhancement, the stimulation of P uptake and P-recycling mechanisms.

Chilling temperatures represent a challenge for crop species originating from warm geographical areas. In this situation, biostimulants serve as an eco-friendly resource to mitigate cold stress in crops. Tomato ( Solanum lycopersicum L.) is an economically important vegetable crop, but quite sensitive to cold stress, which it encounters in both open field and greenhouse settings. In this study, the biostimulant effect of a brown-seaweed extract (BSE) has been evaluated in tomato exposed to low temperature. To assess the product effects, physiological and molecular characterizations were conducted. Under cold stress conditions, stomatal conductance, net photosynthesis, and yield were significantly (p ≤ 0.05) higher in BSE-treated plants compared to the untreated ones. A global transcriptomic survey after BSE application revealed the impact of the BSE treatment on genes leading to key responses to cold stress. This was highlighted by the significantly enriched GO categories relative to proline (GO:0006560), flavonoids (GO:0009812, GO:0009813), and chlorophyll (GO:0015994). Molecular data were integrated by biochemical analysis showing that the BSE treatment causes greater proline, polyphenols, flavonoids, tannins, and carotenoids contents.The study highlighted the role of antioxidant molecules to enhance tomato tolerance to low temperature mediated by BSE-based biostimulant., Competing Interests: FMag is employed by Sipcam Italia S.p.A. belonging together with Sofbey SA to the Sipcam Oxon S.p.A. Group Pero, Italy. FMan is a former employer at Sipcam Italia S.p.A. belonging together with Sofbey SA to the Sipcam Oxon S.p.A. Group Pero, Italy. FMan was employed at Sipcam during the time of the study. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Borella, Baghdadi, Bertoldo, Della Lucia, Chiodi, Celletti, Deb, Baglieri, Zegada-Lizarazu, Pagani, Monti, Mangione, Magro, Hermans, Stevanato and Nardi.)

Late blight, caused by oomycetes Phytophthora infestans is one of the most challenging fungal diseases to manage in tomato plants ( Solanum lycopersicum L.). Toward managing the disease, conventional breeding has successfully introgressed genetic loci conferring disease resistance from various wild relatives of tomato into commercial varieties. The cataloging of disease-associated SNP markers and a deeper understanding of disease-resistance mechanisms are needed to keep up with the demand for commercial varieties resistant against emerging pathogen strains. To this end, we performed transcriptome sequencing to evaluate the gene expression dynamics of tomato varieties, resistant and susceptible to Phytophthora infection. Further integrating the transcriptome dataset with large-scale public genomic data of varieties with known disease phenotypes, a panel of single nucleotide polymorphism (SNP) markers correlated with disease resistance was identified. These SNPs were then validated on 31 lines with contrasting phenotypes for late blight. The identified SNPs are located on genes coding for a putative cysteine-rich transmembrane module (CYSTM), Solyc09g098310, and a nucleotide-binding site-leucine-rich repeat protein, Solyc09g098100, close to the well-studied Ph-3 resistance locus known to have a role in plant immunity against fungal infections. The panel of SNPs generated by this study using transcriptome sequencing showing correlation with disease resistance across a broad set of plant material can be used as markers for molecular screening in tomato breeding.

The molecular and phenotypic effects of a brown seaweed extract (BSE) were assessed in sugar beet ( Beta vulgaris L.). Transcript levels of BSE-treated and untreated plants were studied by RNA-seq and validated by quantitative real-time PCR analysis (RT-qPCR). Root morphology, sugar yield, and processing quality traits were also analyzed to better elucidate the treatment effects. RNA-seq revealed 1019 differentially expressed genes (DEGs) between the BSE-treated and untreated plants. An adjusted p -value < 0.1 and an absolute value of log2 (fold change) greater than one was used as criteria to select the DEGs. Gene ontology (GO) identified hormone pathways as an enriched biological process. Six DEGs involved in auxin and ABA pathways were validated using RT-qPCR. The phenotypic characterization indicated that BSE treatment led to a significant increase ( p < 0.05) in total root length and the length of fine roots of plants grown under hydroponics conditions. The sugar yield of plants grown under field conditions was higher ( p < 0.05) in the treated field plots compared with the control treatment, without impacting the processing quality. Our study unveiled the relevant effects of BSE application in regulating auxin- and ABA-related gene expression and critical traits related to sugar beet development and yield.

A comprehensive approach using phenomics and global transcriptomics for dissecting plant response to biostimulants is illustrated with tomato ( Solanum lycopersicum cv. Micro-Tom and Rio Grande) plants cultivated in the laboratory, greenhouse, and open field conditions. Biostimulant treatment based on an Ascophyllum nodosum extract (ANE) was applied as a foliar spray with two doses (1 or 2 l ha -1 ) at three different phenological stages (BBCH51, BBCH61, and BBCH65) during the flowering phase. Both ANE doses resulted in greater net photosynthesis rate, stomatal conductance, and fruit yield across all culture conditions. A global transcriptomic analysis of leaves from plants grown in the climate chamber, revealed a greater number of differentially expressed genes (DEGs) with the low ANE dose compared to the greater one. The second and third applications induced broader transcriptome changes compared to the first one, indicating a cumulative treatment effect. The functional enrichment analysis of DEGs highlighted pathways related to stimulus-response and photosynthesis, consistent with the morpho-physiological observations. This study is the first comprehensive dual-omics approach for profiling plant responses to biostimulants across three different culture conditions., Competing Interests: Author Francesca Mangione is employed by Sipcam Italia S.p.A. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. No funds from a commercial party were received in support of this article. No benefits in any form have been or will be received from a commercial party related directly or indirectly to the subject of this manuscript. No influence on the interpretation of the data and writing of the article was exerted by a commercial party., (Copyright © 2022 Baghdadi, Della Lucia, Borella, Bertoldo, Ravi, Zegada-Lizarazu, Chiodi, Pagani, Hermans, Stevanato, Nardi, Monti and Mangione.)

The fungus Cercospora beticola causes Cercospora Leaf Spot (CLS) of sugar beet (Beta vulgaris L.). Despite the global importance of this disease, durable resistance to CLS has still not been obtained. Therefore, the breeding of tolerant hybrids is a major goal for the sugar beet sector. Although recent studies have suggested that the leaf microbiome composition can offer useful predictors to assist plant breeders, this is an untapped resource in sugar beet breeding efforts. Using Ion GeneStudio S5 technology to sequence amplicons from seven 16S rRNA hypervariable regions, the most recurring endophytes discriminating CLS-symptomatic and symptomless sea beets (Beta vulgaris L.ssp. maritima) were identified. This allowed the design of taxon-specific primer pairs to quantify the abundance of the most representative endophytic species in large naturally occurring populations of sea beet and subsequently in sugar beet breeding genotypes under either CLS symptomless or infection stages using qPCR. Among the screened bacterial genera, Methylobacterium and Mucilaginibacter were found to be significantly (p < 0.05) more abundant in symptomatic sea beets with respect to symptomless. In cultivated sugar beet material under CLS infection, the comparison between resistant and susceptible genotypes confirmed that the susceptible genotypes hosted higher contents of the above-mentioned bacterial genera. These results suggest that the abundance of these species can be correlated with increased sensitivity to CLS disease. This evidence can further prompt novel protocols to assist plant breeding of sugar beet in the pursuit of improved pathogen resistance., (© 2022. The Author(s).)

The identification of efficient molecular markers related to low bolting tendency is a priority in sugar beet ( Beta vulgaris L.) breeding. This study aimed to identify SNP markers associated with low bolting tendency by establishing a genome-wide association study. An elaborate 3-year field trial comprising 13 sugar beet lines identified L14 as the one exhibiting the lowest bolting tendency along with an increased survival rate after autumnal sowing. For SNP discovery following phenotyping, contrasting phenotypes of 24 non-bolting and 15 bolting plants of the L14 line were sequenced by restriction site-associated DNA sequencing (RAD-seq). An association model was established with a set of 10,924 RAD-based single nucleotide polymorphism (SNP) markers. The allelic status of the most significantly associated SNPs ranked based on their differential allelic status between contrasting phenotypes ( p < 0.01) was confirmed on three different validation datasets comprising diverse sugar beet lines and varieties adopting a range of SNP detection technologies. This study has led to the identification of SNP&#95;36780842 and SNP&#95;48607347 linked to low bolting tendency and can be used for marker-assisted breeding and selection in sugar beet., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ravi, Campagna, Della Lucia, Broccanello, Bertoldo, Chiodi, Maretto, Moro, Eslami, Srinivasan, Squartini, Concheri and Stevanato.)

Sulfur is an essential plant macronutrient, and its adequate supply allows an efficient root storage and sugar extractability in sugar beets ( Beta &nbsp; vulgaris L.). In this study, we investigated the effect of changes in sulfur availability on the endophytic community structure of sugar beets. Plants were hydroponically grown in a complete nutrient solution (S-supplied), a nutrient solution without MgSO 4 (S-deprived), and a nutrient solution without MgSO 4 for six days and resupplied with 100 μM MgSO 4 for 48 h (S-resupplied). The sulfur status was monitored by inductively coupled plasma ICP-OES, and combustion analysis together with the evaluation of microRNA395 as a biomarker for sulfate status. Metabarcoding of the bacterial 16S rRNA gene was carried out in order to determine leaf endophytic community structure. The Shannon diversity index significantly differed ( p < 0.05) between sulfate-supplied and sulfate-deprived seedlings. Validation by Real-Time PCR showed a significant increase ( p < 0.05) of Burkholderia spp. in sulfate-deprived plants as compared to sulfate-supplied ones. The study sheds new light on the effects of nutrient deficiency on the microbiome of sugar beet plants.

The present study aimed to explore the effects of foliar application of a leonardite-based product on sugar beet ( Beta vulgaris L.) plants grown in the field. The approach concerned the evaluation of the community compositional structure of plant endophytic bacteria through a metabarcoding approach, the expression level of a gene panel related to hormonal metabolism and signaling, and the main sugar beet productivity traits. Results indicated that plants treated with leonardite (dosage of 2,000 ml ha -1 , dilution 1:125, 4 mg C l -1 ) compared with untreated ones had a significant increase ( p < 0.05) in (i) the abundance of Oxalicibacterium spp., recognized to be an endophyte bacterial genus with plant growth-promoting activity; (ii) the expression level of LAX2 gene, coding for auxin transport proteins; and (iii) sugar yield. This study represents a step forward to advance our understanding of the changes induced by leonardite-based biostimulant in sugar beet., Competing Interests: FM was employed by company Sipcam Oxon S.p.A. GCa was employed by company COPROB. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Della Lucia, Bertoldo, Broccanello, Maretto, Ravi, Marinello, Sartori, Marsilio, Baglieri, Romano, Colombo, Magro, Campagna, Concheri, Squartini and Stevanato.)

Prospect theory is among the most influential frameworks in behavioural science, specifically in research on decision-making under risk. Kahneman and Tversky's 1979 study tested financial choices under risk, concluding that such judgements deviate significantly from the assumptions of expected utility theory, which had remarkable impacts on science, policy and industry. Though substantial evidence supports prospect theory, many presumed canonical theories have drawn scrutiny for recent replication failures. In response, we directly test the original methods in a multinational study (n = 4,098 participants, 19 countries, 13 languages), adjusting only for current and local currencies while requiring all participants to respond to all items. The results replicated for 94% of items, with some attenuation. Twelve of 13 theoretical contrasts replicated, with 100% replication in some countries. Heterogeneity between countries and intra-individual variation highlight meaningful avenues for future theorizing and applications. We conclude that the empirical foundations for prospect theory replicate beyond any reasonable thresholds.

In the past two decades, psychological science has experienced an unprecedented replicability crisis, which has uncovered several issues. Among others, the use and misuse of statistical inference plays a key role in this crisis. Indeed, statistical inference is too often viewed as an isolated procedure limited to the analysis of data that have already been collected. Instead, statistical reasoning is necessary both at the planning stage and when interpreting the results of a research project. Based on these considerations, we build on and further develop an idea proposed by Gelman and Carlin (2014) termed "prospective and retrospective design analysis." Rather than focusing only on the statistical significance of a result and on the classical control of type I and type II errors, a comprehensive design analysis involves reasoning about what can be considered a plausible effect size. Furthermore, it introduces two relevant inferential risks: the exaggeration ratio or Type M error (i.e., the predictable average overestimation of an effect that emerges as statistically significant) and the sign error or Type S error (i.e., the risk that a statistically significant effect is estimated in the wrong direction). Another important aspect of design analysis is that it can be usefully carried out both in the planning phase of a study and for the evaluation of studies that have already been conducted, thus increasing researchers' awareness during all phases of a research project. To illustrate the benefits of a design analysis to the widest possible audience, we use a familiar example in psychology where the researcher is interested in analyzing the differences between two independent groups considering Cohen's d as an effect size measure. We examine the case in which the plausible effect size is formalized as a single value, and we propose a method in which uncertainty concerning the magnitude of the effect is formalized via probability distributions. Through several examples and an application to a real case study, we show that, even though a design analysis requires significant effort, it has the potential to contribute to planning more robust and replicable studies. Finally, future developments in the Bayesian framework are discussed., (Copyright © 2020 Altoè, Bertoldo, Zandonella Callegher, Toffalini, Calcagnì, Finos and Pastore.)

Leonardite-based biostimulants are a large class of compounds, including humic acid substances. Foliar application of biostimulants at field level improves plant growth, yield and quality through metabolic changes and stimulation of plant proton pumps. The present study aimed at identifying optimum dosage of BLACKJAK, a humic acid-based substance, which is able to modify genes involved in sugar beet growth. Thirty-three genes belonging to various biochemical pathway categories were tested in leaves of treated sugar beet ( Beta vulgaris L.) samples to assess gene expression profiling in response to BLACKJAK. Seedlings of a diploid and multigerm variety were grown in plastic pots and sprayed with two dilutions of BLACKJAK (dilution 1:500-1.0 mg C L -1 and dilution 1:1000-0.5 mg C L -1 ). Leaf samples were collected after 24, 48, and 72 h treatment with BLACKJAK for each dilution. RNA was extracted and the quantification of gene expression was performed while using an OpenArray platform. Results of analysis of variance demonstrated that, 15 genes out of a total of 33 genes tested with OpenArray qPCR were significantly affected by treatment and exposure time. Analysis for annotation of gene products and pathways revealed that genes belonging to the mitochondrial respiratory pathways, nitrogen and hormone metabolisms, and nutrient uptake were up-regulated in the BLACKJAK treated samples. Among the up-regulated genes, Bv&#95;PHT2;1 and Bv&#95;GLN1 expression exerted a 2-fold change in 1:1000 and 1:500 BLACKJAK concentrations. Overall, the gene expression data in the BLACKJAK treated leaves demonstrated the induction of plant growth-related genes that were contributed almost to amino acid and nitrogen metabolism, plant defense system, and plant growth.

Humic substances extracted from leonardite are widely considered to be bioactive compounds, influencing the whole-plant physiology and the crop yield. The aim of this work was to evaluate the effect of a new formulate based on leonardite in the early stage of growth of sugar beet ( Beta vulgaris L.). A commercial preparation of leonardite (BLACKJAK) was characterized by ionomic analysis, solid-state 13 C MAS NMR spectroscopy. Seedlings of sugar beet were grown in Hoagland's solution under controlled conditions. After five days of growth, an aliquot of the concentrated BLACKJAK was added to the solution to obtain a final dilution of 1:1000 (0.5 mg C L -1 ). The sugar beet response in the early stage of growth was determined by evaluating root morphological traits as well as the changes in the expression of 53 genes related to key morphophysiological processes. Root morphological traits, such as total root length, fine root length (average diameter < 0.5 mm), and number of root tips, were significantly ( p < 0.001) increased in plants treated with BLACKJAK, compared to the untreated plants at all sampling times. At the molecular level, BLACKJAK treatment upregulated many of the evaluated genes. Moreover, both Real Time PCR and digital PCR showed that genes involved in hormonal response, such as PIN, ARF3, LOGL 10, GID1, and BRI1, were significantly ( p < 0.05) upregulated by treatment with BLACKJAK. Our study provides essential information to understand the effect of a leonardite-based formulate on plant growth hormone metabolism, although the molecular and physiological basis for these complicated regulatory mechanisms deserve further investigations., Competing Interests: The authors declare no conflict of interest.

This cross-sectional study aimed at evaluating the knowledge and attitudes concerning influenza vaccination in Southern Italy, and investigating the potential determinants of vaccine uptake. The sample consisted of 700 adults (mean age 58.7y) with chronic diseases attending four public specialty clinics in Italy. Overall, 64.7% of the participants were aware that influenza can be prevented with vaccines and that patients with chronic diseases are at higher risk of developing severe complications. Less than half of the sample (42.1%) received influenza vaccine in the last season, and 46.9% declared the will to receive influenza vaccination in the next season. The level of awareness was significantly lower among the elderly (> = 65y) and those with a higher self-reported health. A significantly higher likelihood of vaccination was observed among the elderly, the subjects with a higher knowledge about vaccine utility and safety, the participants with chronic respiratory diseases, and those who had taken more drugs. Future education programs and communication strategies are strongly needed in adults with chronic diseases to improve influenza vaccination knowledge and uptake., Competing Interests: The authors have declared that no competing interests exist.

In this study, a system based on omics profiling was set-up for sugar beet ( Beta vulgaris L. subsp. vulgaris ) evaluation after changes in sulfate availability. Seedlings were grown on sulfate-deprived Hoagland solution. Six days after germination, 100 μM MgSO 4 was added to the solution. Root samples were collected 36 h after treatments. WinRHIZO root-scanning approach was used for the automated image analysis of plant root morphology. Inductively Coupled Plasma Spectrometry (ICP-OES) and quadrupole-time-of-flight mass spectrometry (Q-TOF) were used for ionomic and metabolic analysis, respectively. Nanofluidic real-time PCR (OpenArray system) was used for molecular profiling. OpenArray chips were designed with TaqMan probes for 53 sugar beet genes putatively involved in sulfate nutrition. At morphological level treated seedlings showed significantly higher values ( P < 0.01) than untreated plants for root traits related to soil exploration and nutrient uptake, such as total root length, fine roots length and root tips number. ICP-OES, Q-TOF and transcriptomic data revealed changes due to sulfate availability in sugar beet samples. Two key results are highlighted in sulfate-supplied roots and leaves. Firstly, high expression levels of auxin efflux carrier component 1 (PIN) and 5-phosphoribosyl-anthranilate, precursor of tryptophan and auxin synthesis, were observed in roots. Secondly, high levels of 2-Cys peroxiredoxin BAS1, chloroplastic, thioredoxin reductase (NADPH) and cysteine synthase, chloroplastic/chromoplastic, O- acetylserine sulfhydrylase, involved in protection against oxidative stress and cysteine synthase activity, respectively, were observed in leaves. Based on our findings, the combination of evaluated omics approaches could become a key system for the evaluation of the nutritional status of sugar beet under different nutrient availability conditions.

Mite infestations to the culture of mate-tea frequently causes losses by the premature fall of the leaves. So, it is necessary to monitor the population of these arthropods, and to adopt management strategies for their control. The objective of this research was to evaluate the trustworthiness of presence-absence sampling for Oligonychus yothersi (McGregor) on mate-tea orchard. This study was conducted in Cascavel, Paraná State, from April 2001 to July 2000, in a mate-tea tree commercial plantation. Biweekly sampling of 240 leaves were collected in different parts of 10 plants, and the number of mites was counted. The aggregation pattern was determined through the coefficients a and b of Taylor's power law. The proportion of infested leaves and the number of required samples were estimated through mathematical model. The mite O. yothersi presented aggregate distribution. The proportion of infested leaves calculated by means of mathematical model showed to be a trustworthy parameter to estimate the population density of the mite. The number of required samples was small, making feasible the practical application of the presence-absence sampling method for O. yothersi in the culture of mate-tea.

Purpose: More than ten years after ETDRS, Optical Coherence Tomography (OCT) greatly enhanced our ability to detect macular thickening and has brought new insights on the morphology of edema and on the presence of vitreal traction. In this study we propose a new classification of macular edema based on OCT findings to better catalogue and follow this complex clinical entity., Methods: Since January 2000 we analysed with OCT 2 (Zeiss Inc.) more than one thousand and two hundred eyes with DME., Results: The classification takes into account five parameters: retinal thickness, diffusion, volume, morphology and presence of vitreous traction. Standard figures and numerical values for every parameter are given., Conclusion: Although ETDRS guidelines for laser treatment of DME still remain the only proven therapy for this condition, many other strategies are now on trial, and the vast majority of authors use OCT as the best indicator of therapeutic benefit. The amount of information given by OCT demonstrates that macular edema is a complex clinical entity with various morphology and gravity, and disclaimed the limitations of a simple "clinical" definition. As in many other examples such as macular holes and choroidal neovascularization, a uniform and precise definition of macular edema would increase the possibility to compare and judge the result of different therapeutic strategies. Aim of this classification is to implement the ETDRS clinical definition of DME with the precise and useful data given by OCT to better diagnose, catalogue and follow macular edema.

Purpose: To determine the effectiveness and safety of diode laser, pars plana vitrectomy (PPV) with gas tamponade, and intravitreal triamcinolone (IVT) as possible treatments for diffuse diabetic macular edema (dDME). To determine whether the new macular edema Classification previously proposed by some of the authors may help as a guide in the choice of treatment., Methods: A retrospective, comparative study of 169 eyes with dDME that underwent treatment. The eyes divided into 3 groups: in the PPV Group, 59 eyes received PPV with gas tamponade; in the DIODE Group, 53 eyes received a laser grid; in the IVT Group, 57 eyes received an intravitreal injection of 4 mg of triamcinolone acetonide. The follow up ranged from 6 to 24 months. The eyes were classified according to the new DME Classification based on OCT., Main Outcomes: Change in foveal thickness as determined by Optical Coherence Tomography (OCT); change in visual acuity; intra and postoperative complications., Results: Mean visual acuity (VA) improved at 3 months with every treatment. At one year only eyes which underwent PPV still had better VA than the pre-op value, while eyes which underwent IVT and laser treatment showed regression. PPV show the greater percentage of eyes which gain 3 or more lines of VA at one year (17%). Mean foveal thickness improved at 3 months with every treatment. Only the eyes into the PPV Group showed relatively low foveal thickness at one year. Better final VA and foveal thickness were obtained if preop VA is > or = 0.3 and if earlier stages of DME were treated according to the new DME Classification. PPV was the treatment which offered the most stable results with at one year or longer. Similar results were observed into the PPV Groups as a whole and into a subgroup of eyes with preoperative PVD. No complications were encountered with laser diode treatment. Long term complications into the IVT Group were elevated IOP (8%), retinal detachment (3.5%) and posterior cataract (15%). Long term complications into the PPV Group were retinal detachment (3.4%) and cataract (90%)., Conclusions: Diode laser, PPV with gas tamponade and IVT are effective alternative treatments to decrease foveal thickness and improve visual acuity in eyes with DME. However while the results of PPV are stable in the long term follow-up, diode laser and IVT do not offer stable results. Complications may be severe with PPV and IVT. It is necessary to carefully select cases which would benefit from these types of treatments. The authors think that the OCT Classification may serve as a guide for the choice of treatment.