East Timor as an important source of cashew (Anacardium occidentale L.) genetic diversity.

Background: Cashew (Anacardium occidentale L.) is a crop currently grown in several tropical countries because of the economic importance of cashew nuts. Despite its enormous economic worth, limited research has been conducted on the molecular diversity of cashew genetic resources. In this study, a wide comprehensive assessment of the genetic diversity of cashew trees in East Timor was performed using microsatellites (SSRs) to evaluate intraspecific diversity and population structuring. Methods: A total of 207 individual cashew trees, including trees from East Timor (11), and outgroup populations from Indonesia (one) and Mozambique (two), were analyzed with 16 cashew-specific SSRs. A comprehensive sampling of cashew trees within East Timor was performed, covering the distribution of cashew orchards in the country. Genetic diversity indices were calculated, and population structuring was determined using three different approaches: genetic distances (UPGMA and NJ), AMOVA, and individual-based clustering methods through Bayesian (STRUCTURE) and multivariate (DAPC) analyses. Results: The population structuring analysis revealed that the genetic diversity of cashew populations in East Timor was higher in this study than previously reported for cashew trees. A higher allelic richness was found within cashew populations in East Timor compared with the outgroup populations (Mozambique and Indonesia), reinforced by the presence of private alleles. Moreover, our study showed that cashew populations in East Timor are grouped into two dissimilar genetic groups, which may suggest multiple cashew introductions over time. These new cashew genetic resources could be explored for future crop improvement. Conclusions: Crop diversity underpins the productivity, resilience, and adaptive capacity of agriculture. Therefore, this study provides useful information regarding genetic diversity and population structure that can be harnessed to improve cashew production in East Timor. This data is also important to creating a country-specific genetic cashew signature to increase cashew market value.

Umbilical-Cord-Derived Mesenchymal Stromal Cells Modulate 26 Out of 41 T Cell Subsets from Systemic Sclerosis Patients.

Systemic sclerosis (SSc) is an immune-mediated disease wherein T cells are particularly implicated, presenting a poor prognosis and limited therapeutic options. Thus, mesenchymal-stem/stromal-cell (MSC)-based therapies can be of great benefit to SSc patients given their immunomodulatory, anti-fibrotic, and pro-angiogenic potential, which is associated with low toxicity. In this study, peripheral blood mononuclear cells from healthy individuals (HC, n = 6) and SSc patients (n = 9) were co-cultured with MSCs in order to assess how MSCs affected the activation and polarization of 58 different T cell subsets, including Th1, Th17, and Treg. It was found that MSCs downregulated the activation of 26 out of the 41 T cell subsets identified within CD4+, CD8+, CD4+CD8+, CD4-CD8-, and γδ T cells in SSc patients (HC: 29/42) and affected the polarization of 13 out of 58 T cell subsets in SSc patients (HC: 22/64). Interestingly, SSc patients displayed some T cell subsets with an increased activation status and MSCs were able to downregulate all of them. This study provides a wide-ranging perspective of how MSCs affect T cells, including minor subsets. The ability to inhibit the activation and modulate the polarization of several T cell subsets, including those implicated in SSc's pathogenesis, further supports the potential of MSC-based therapies to regulate T cells in a disease whose onset/development may be due to immune system's malfunction.

Lemierre's Syndrome: A Case Report.

Lemierre's syndrome is a rare condition characterized by septic thrombophlebitis of the internal jugular vein secondary to infection of the head and neck region and septic embolization to other organs. The most frequent etiological agent is Fusobacterium necrophorum, a commensal anaerobic gram-negative bacillus of the oral flora. We report the case of a young male who presented with chest pain after a dental procedure. He developed a masseterian phlegmon, thrombosis of the internal jugular vein, and embolization to the lung complicated by empyema. The diagnosis of Lemierre's syndrome was delayed by the negative blood cultures, but full recovery was achieved after appropriate broad-spectrum antibiotic coverage. Our main objective is to highlight the fact that a high clinical suspicion is required to establish the diagnosis of this rare syndrome.

O que mudou entre os períodos de pico e de platô durante a primeira onda do SARS-CoV-2? Estudo multicêntrico português em unidades de cuidados intensivos / What changed between the peak and plateau periods of the first COVID-19 pandemic wave? A multicentric Portuguese cohort study in intensive care

RESUMO Objetivo: Analisar e comparar as características de pacientes críticos com a COVID-19, a abordagem clínica e os resultados entre os períodos de pico e de platô na primeira onda pandêmica em Portugal. Métodos: Este foi um estudo de coorte multicêntrico ambispectivo, que incluiu pacientes consecutivos com a forma grave da COVID-19 entre março e agosto de 2020 de 16 unidades de terapia intensiva portuguesas. Definiram-se as semanas 10 - 16 e 17 - 34 como os períodos de pico e platô. Resultados: Incluíram-se 541 pacientes adultos com mediana de idade de 65 [57 - 74] anos, a maioria do sexo masculino (71,2%). Não houve diferenças significativas na mediana de idade (p = 0,3), no Simplified Acute Physiology Score II (40 versus 39; p = 0,8), na pressão parcial de oxigênio/fração inspirada de oxigênio (139 versus 136; p = 0,6), na terapia com antibióticos na admissão (57% versus 64%; p = 0,2) ou na mortalidade aos 28 dias (24,4% versus 22,8%; p = 0,7) entre o período de pico e platô. Durante o período de pico, os pacientes tiveram menos comorbidades (1 [0 - 3] versus 2 [0 - 5]; p = 0,002); fizeram mais uso de vasopressores (47% versus 36%; p < 0,001) e ventilação mecânica invasiva na admissão (58,1% versus 49,2%; p < 0,001), e tiveram mais prescrição de hidroxicloroquina (59% versus 10%; p < 0,001), lopinavir/ritonavir (41% versus 10%; p < 0,001) e posição prona (45% versus 36%; p = 0,04). Entretanto, durante o platô, observou-se maior uso de cânulas nasais de alto fluxo (5% versus 16%; p < 0,001) na admissão, remdesivir (0,3% versus 15%; p < 0,001) e corticosteroides (29% versus 52%; p < 0,001), além de menor tempo de internação na unidade de terapia intensiva (12 versus 8 dias; p < 0,001). Conclusão: Houve mudanças significativas nas comorbidades dos pacientes, nos tratamentos da unidade de terapia intensiva e no tempo de internação entre os períodos de pico e platô na primeira onda da COVID-19.

ABSTRACT Objective: To analyze and compare COVID-19 patient characteristics, clinical management and outcomes between the peak and plateau periods of the first pandemic wave in Portugal. Methods: This was a multicentric ambispective cohort study including consecutive severe COVID-19 patients between March and August 2020 from 16 Portuguese intensive care units. The peak and plateau periods, respectively, weeks 10 - 16 and 17 - 34, were defined. Results: Five hundred forty-one adult patients with a median age of 65 [57 - 74] years, mostly male (71.2%), were included. There were no significant differences in median age (p = 0.3), Simplified Acute Physiology Score II (40 versus 39; p = 0.8), partial arterial oxygen pressure/fraction of inspired oxygen ratio (139 versus 136; p = 0.6), antibiotic therapy (57% versus 64%; p = 0.2) at admission, or 28-day mortality (24.4% versus 22.8%; p = 0.7) between the peak and plateau periods. During the peak period, patients had fewer comorbidities (1 [0 - 3] versus 2 [0 - 5]; p = 0.002) and presented a higher use of vasopressors (47% versus 36%; p < 0.001) and invasive mechanical ventilation (58.1 versus 49.2%; p < 0.001) at admission, prone positioning (45% versus 36%; p = 0.04), and hydroxychloroquine (59% versus 10%; p < 0.001) and lopinavir/ritonavir (41% versus 10%; p < 0.001) prescriptions. However, a greater use of high-flow nasal cannulas (5% versus 16%, p < 0.001) on admission, remdesivir (0.3% versus 15%; p < 0.001) and corticosteroid (29% versus 52%, p < 0.001) therapy, and a shorter ICU length of stay (12 days versus 8, p < 0.001) were observed during the plateau. Conclusion: There were significant changes in patient comorbidities, intensive care unit therapies and length of stay between the peak and plateau periods of the first COVID-19 wave.

Diversity of Legumes in the Cashew Agroforestry System in East Timor (Southeast Asia).

Cashew is an important export-oriented crop in several tropical countries, often under monocropping systems. Intercropping with legume species is promoted as a sustainable practice, enhancing agricultural productivity and providing nutritional food sources to rural communities. This study aimed to characterize the diversity of Leguminosae (or Fabaceae) in the cashew agroforestry systems of East Timor (Southeast Asia). Fourteen cashew orchards were sampled across the country, and information about leguminous species uses was collected from local populations. About 50 species are commonly part of the country's cashew agroforestry system, many of them simultaneously used as food, fodder, and in traditional medicine. Six bean species-Cajanus cajan (L.) Huth, Phaseolus lunatus L., Phaseolus vulgaris L., Vigna angularis (Willd.) Ohwi and H.Ohashi, Vigna radiata (L.) R.Wilczek and Vigna unguiculata (L.) Walp.-are largely used as food. The mineral contents of these beans revealed relevant differences between species and, in some cases, between types (seed colour) within species. Periods of hunger and low food variety are frequent in East Timor, reflecting a very poor nutritional state of the population. Knowing and using legumes for local nutrition, as well as for healthcare and well-being, adds great value to these species as components of East Timor cashew agroforestry systems.

Tuberculosis and Its "Troubled Relationship" With Other Diseases.

Tuberculosis (TB) is a multisystemic disease caused most frequently by Mycobacterium tuberculosis. Extrapulmonary TB has become more frequent with the advent of human immunodeficiency virus (HIV) as HIV can facilitate the infection with M. tuberculosis, especially during HIV seroconversion. Here, we present the case of a 22-year-old man, from Guinea-Bissau, with a history of untreated HIV who was admitted to the intensive care unit for respiratory failure needing mechanical ventilation. Pulmonary TB was diagnosed. His stay was complicated with a hemorrhagic shock due to traumatic urethral catheterization, which led to a perforation of the capsule of the prostate. A prostatectomy was needed for bleeding control. The anatomopathological examination confirmed the presence of acid-resistant bacilli, and an extensive caseous type necrosis of the whole tissue, thus diagnosing a prostatic tuberculosis. The patient recovered after a hemorrhagic shock, a urologic and radical intervention, and some severe infectious complications.

Active Neutralizing Mats for Corrosive Chemical Storage.

Laboratories and industries that handle chemicals are ubiquitously prone to leakages. These may occur in storage rooms, cabinets or even in temporary locations, such as workbenches and shelves. A relevant number of these chemicals are corrosive, thus commercial products already exist to prevent material damage and injuries. One strategy consists of the use of absorbing mats, where few display neutralizing properties, and even less a controlled neutralization. Nevertheless, to the authors' knowledge, the commercially available neutralizing mats are solely dedicated to neutralizing acid or alkali solutions, never both. Therefore, this work describes the development and proof of a completely novel concept, where a dual component active mat (DCAM) is able to perform a controlled simultaneous neutralization of acid and alkali leakages by using microencapsulated active components. Moreover, its active components comprise food-grade ingredients, embedded in nonwoven polypropylene. The acid neutralizing mats contain sodium carbonate (Na2CO3) encapsulated in sodium alginate microcapsules (MC-ASC). Alkali neutralizing mats possess commercial encapsulated citric acid in hydrogenated palm oil (MIRCAP CT 85-H). A DCAM encompasses both MC-ASC and MIRCAP CT 85-H and was able to neutralize solutions up to 10% (v/v) of hydrochloric acid (HCl) and sodium hydroxide (NaOH). The efficacy of the neutralization was assessed by direct titration and using pH strip measurement tests to simulate the leakages. Due to the complexity of neutralization efficacy evaluation based solely on pH value, a thorough conductivity study was performed. DCAM reduced the conductivity of HCl and NaOH (1% and 2% (v/v)) in over 70%. The composites were characterized by scanning electron microscopy (SEM), differential calorimetry (DSC) and thermogravimetric analysis (TGA). The size of MC-ASC microcapsules ranged from 2 µm to 8 µm. Finally, all mat components displayed thermal stability above 150 °C.

What changed between the peak and plateau periods of the first COVID-19 pandemic wave? A multicentric Portuguese cohort study in intensive care. / O que mudou entre os períodos de pico e de platô durante a primeira onda do SARS-CoV-2? Estudo multicêntrico português em unidades de cuidados intensivos.

OBJECTIVE: To analyze and compare COVID-19 patient characteristics, clinical management and outcomes between the peak and plateau periods of the first pandemic wave in Portugal. METHODS: This was a multicentric ambispective cohort study including consecutive severe COVID-19 patients between March and August 2020 from 16 Portuguese intensive care units. The peak and plateau periods, respectively, weeks 10 - 16 and 17 - 34, were defined. RESULTS: Five hundred forty-one adult patients with a median age of 65 [57 - 74] years, mostly male (71.2%), were included. There were no significant differences in median age (p = 0.3), Simplified Acute Physiology Score II (40 versus 39; p = 0.8), partial arterial oxygen pressure/fraction of inspired oxygen ratio (139 versus 136; p = 0.6), antibiotic therapy (57% versus 64%; p = 0.2) at admission, or 28-day mortality (24.4% versus 22.8%; p = 0.7) between the peak and plateau periods. During the peak period, patients had fewer comorbidities (1 [0 - 3] versus 2 [0 - 5]; p = 0.002) and presented a higher use of vasopressors (47% versus 36%; p < 0.001) and invasive mechanical ventilation (58.1 versus 49.2%; p < 0.001) at admission, prone positioning (45% versus 36%; p = 0.04), and hydroxychloroquine (59% versus 10%; p < 0.001) and lopinavir/ritonavir (41% versus 10%; p < 0.001) prescriptions. However, a greater use of high-flow nasal cannulas (5% versus 16%, p < 0.001) on admission, remdesivir (0.3% versus 15%; p < 0.001) and corticosteroid (29% versus 52%, p < 0.001) therapy, and a shorter ICU length of stay (12 days versus 8, p < 0.001) were observed during the plateau. CONCLUSION: There were significant changes in patient comorbidities, intensive care unit therapies and length of stay between the peak and plateau periods of the first COVID-19 wave.

OBJETIVO: Analisar e comparar as características de pacientes críticos com a COVID-19, a abordagem clínica e os resultados entre os períodos de pico e de platô na primeira onda pandêmica em Portugal. MÉTODOS: Este foi um estudo de coorte multicêntrico ambispectivo, que incluiu pacientes consecutivos com a forma grave da COVID-19 entre março e agosto de 2020 de 16 unidades de terapia intensiva portuguesas. Definiram-se as semanas 10 - 16 e 17 - 34 como os períodos de pico e platô. RESULTADOS: Incluíram-se 541 pacientes adultos com mediana de idade de 65 [57 - 74] anos, a maioria do sexo masculino (71,2%). Não houve diferenças significativas na mediana de idade (p = 0,3), no Simplified Acute Physiology Score II (40 versus 39; p = 0,8), na pressão parcial de oxigênio/fração inspirada de oxigênio (139 versus 136; p = 0,6), na terapia com antibióticos na admissão (57% versus 64%; p = 0,2) ou na mortalidade aos 28 dias (24,4% versus 22,8%; p = 0,7) entre o período de pico e platô. Durante o período de pico, os pacientes tiveram menos comorbidades (1 [0 - 3] versus 2 [0 - 5]; p = 0,002); fizeram mais uso de vasopressores (47% versus 36%; p < 0,001) e ventilação mecânica invasiva na admissão (58,1% versus 49,2%; p < 0,001), e tiveram mais prescrição de hidroxicloroquina (59% versus 10%; p < 0,001), lopinavir/ritonavir (41% versus 10%; p < 0,001) e posição prona (45% versus 36%; p = 0,04). Entretanto, durante o platô, observou-se maior uso de cânulas nasais de alto fluxo (5% versus 16%; p < 0,001) na admissão, remdesivir (0,3% versus 15%; p < 0,001) e corticosteroides (29% versus 52%; p < 0,001), além de menor tempo de internação na unidade de terapia intensiva (12 versus 8 dias; p < 0,001). CONCLUSÃO: Houve mudanças significativas nas comorbidades dos pacientes, nos tratamentos da unidade de terapia intensiva e no tempo de internação entre os períodos de pico e platô na primeira onda da COVID-19.

Nitrogen Acquisition and Transport in the Ectomycorrhizal Symbiosis-Insights from the Interaction between an Oak Tree and Pisolithus tinctorius.

In temperate forests, the roots of various tree species are colonized by ectomycorrhizal fungi, which have a key role in the nitrogen nutrition of their hosts. However, not much is known about the molecular mechanisms related to nitrogen metabolism in ectomycorrhizal plants. This study aimed to evaluate the nitrogen metabolic response of oak plants when inoculated with the ectomycorrhizal fungus Pisolithus tinctorius. The expression of candidate genes encoding proteins involved in nitrogen uptake and assimilation was investigated in ectomycorrhizal roots. We found that three oak ammonium transporters were over-expressed in root tissues after inoculation, while the expression of amino acid transporters was not modified, suggesting that inorganic nitrogen is the main form of nitrogen transferred by the symbiotic fungus into the roots of the host plant. Analysis by heterologous complementation of a yeast mutant defective in ammonium uptake and GFP subcellular protein localization clearly confirmed that two of these genes encode functional ammonium transporters. Structural similarities between the proteins encoded by these ectomycorrhizal upregulated ammonium transporters, and a well-characterized ammonium transporter from E. coli, suggest a similar transport mechanism, involving deprotonation of NH4+, followed by diffusion of uncharged NH3 into the cytosol. This view is supported by the lack of induction of NH4+ detoxifying mechanisms, such as the GS/GOGAT pathway, in the oak mycorrhizal roots.

COVID-19-Associated Invasive Pulmonary Aspergillosis in the Intensive Care Unit: A Case Series in a Portuguese Hospital.

Invasive pulmonary aspergillosis (IPA) has become a recognizable complication in coronavirus disease 2019 (COVID-19) patients admitted to intensive care units (ICUs). Alveolar damage in the context of acute respiratory distress syndrome (ARDS) appears to be the culprit in facilitating fungal invasion in COVID-19 patients, leading to a COVID-19-associated pulmonary aspergillosis (CAPA) phenomenon. From November 2020 to 15 February 2021, 248 COVID-19 patients were admitted to our ICUs, of whom ten patients (4% incidence) were classified as either probable (six) or possible (four) CAPA cases. Seven patients had positive cultural results: Aspergillus fumigatus sensu stricto (five), A. terreus sensu stricto (one), and A. welwitschiae (one). Five patients had positive bronchoalveolar lavage (BAL) and galactomannan (GM), and two patients had both positive cultural and GM criteria. All but two patients received voriconazole. Mortality rate was 30%. Strict interpretation of classic IPA definition would have resulted in eight overlooked CAPA cases. Broader diagnostic criteria are essential in this context, even though differentiation between Aspergillus colonization and invasive disease might be more challenging. Herein, we aim to raise awareness of CAPA in view of its potential detrimental outcome, emphasizing the relevance of a low threshold for screening and early antifungal treatment in ARDS patients.

Traumatic Isolated Thumb Carpometacarpal Joint Dislocation - Report of Two Clinical Cases / Luxação traumática isolada da articulação trapéziometacárpica - Relato de dois casos clínicos

Abstract Isolated thumb carpometacarpal joint dislocation is a rare lesion that accounts for less than 1% of all hand lesions. The authors present two cases of traumatic isolated thumb carpometacarpal joint dislocation. One of them was treated with closed reduction and cast immobilization, and the other was treated with closed reduction, Kirschner-wires pinning, and cast immobilization. The first patient had a good functional outcome and showed no signs of thumb carpometacarpal instability. The patient treated with Kirschner wires presented signs of clinical instability and radiological subluxation. Isolated thumb carpometacarpal dislocation is a rare lesion that can cause joint instability, which interferes with the normal function of the hand and can lead to articular degenerative changes. The best management of this lesion is still controversial, since there is lack of evidence in the literature showing superiority of one treatment over the other.

Resumo A luxação traumática isolada da articulação trapézio-metacárpica é uma lesão rara que faz parte de menos de 1% de todas as lesões de mãos. Os autores apresentam dois casos de luxação traumática isolada da articulação trapézio-metacárpica. Um dos casos foi tratado com redução fechada e imobilização com gesso, e o outro foi tratado com redução fechada, fixação com fios Kirschner, e imobilização com gesso. O primeiro paciente teve um bom resultado funcional e não mostrou sinais de instabilidade trapeziometacarpal. O paciente tratado com fios Kirschner apresentou sinais de instabilidade clínica e subluxação radiológica. A luxação isolada da articulação trapeziometacarpal é uma lesão rara que pode causar instabilidade articular que interfere com a funcionalidade normal da mão e pode resultar em mudanças articulares degenerativas. O melhor manejo dessa lesão ainda é controverso, já que ainda faltam evidências na literatura que mostrem a superioridade de um tratamento em relação ao outro.

The Reality of Critical Cancer Patients in a Polyvalent Intensive Care Unit.

Background and objective With the increasing incidence of cancer and the rise in the survival rates of cancer patients, more and more oncological candidates are being considered for admission to intensive care units (ICU). Several studies have demonstrated no difference in the outcomes of cancer patients compared to non-cancer patients. Our study aimed to describe and analyze the outcomes related to cancer patients in a polyvalent ICU. Methods We conducted a retrospective study of consecutive oncological patients admitted to a polyvalent ICU (2013-2017). Cox model and receiver operating characteristic (ROC) curve analysis were performed to analyze the results. Results A total of 236 patients were included in the study; the mean age of the patients was 53.5 ± 15.3 years, and 65% of them were male. The main cancer types were those related to the central nervous system (CNS; 31%), as well as gastrointestinal (18%), genitourinary (17%), and hematological (15%). Curative/diagnostic surgeries (49%) and sepsis/septic shock (17%) were the main reasons for admission. The Acute Physiology and Chronic Health Evaluation II (APACHE II) and Simplified Acute Physiology Score II (SAPS II) scores in hematological patients vs. solid tumors were as follows: 30 vs. 20 and 63 vs. 38, respectively (p<0.005). Vasopressors, invasive mechanical ventilation (IMV), and renal replacement therapy (RRT) were used more widely in hematological patients compared to solid-tumor patients. Length of stay was longer in hematological patients vs. solid-tumor patients (12.8 vs. 7 days, p=0.002). The median overall survival in hematological patients was one month and that in solid-tumor patients was 5.8 months (p<0.005). The survival rate at six months was better than described in the existing literature (48 vs. 32.4%). Conclusion Both SAPS II and APACHE II scores were reasonably accurate in predicting mortality, demonstrating their value in cancer patients.

Tackling Food Insecurity in Cabo Verde Islands: The Nutritional, Agricultural and Environmental Values of the Legume Species.

Legume species are important food sources to reduce hunger and deal with malnutrition; they also play a crucial role in sustainable agriculture in the tropical dry islands of Cabo Verde. To improve the knowledge of the heritage of plant genetic resources in this Middle Income Country, this study had three main goals: (i) to provide a checklist of food legumes; (ii) to investigate which species are traded in local markets and, based on field surveys, to compare species for their chemical, phenolic, antioxidant, and nutritional composition; and (iii) to discuss the agronomic value and contribution to food security in this archipelago. Our results revealed that 15 species are used as food and 5 of them are locally traded (Cajanus cajan, Lablab purpureus, Phaseolus lunatus, Phaseolus vulgaris, and Vigna unguiculata). The role of these species as sources of important minerals, antioxidants, and nutritional components for food security is highlighted, and the native ones (Lablab purpureus and Vigna unguiculata) stand-out as particularly well-adapted to the climate of these islands, which are already experiencing the adverse effects of climate change. We conclude that the sustainable use of these genetic resources can contribute to the reduction of hunger and poverty, thus meeting some challenges of the Sustainable Development Goals.

First Report of Dieback Caused by Neofusicoccum batangarum in Cashew in Guinea-Bissau.

Cashew (Anacardium occidentale L.) is a cash crop with a highly significant economic importance in West Africa, particularly in Guinea-Bissau (Monteiro et al. 2015, 2017). In October 2018, dieback-like symptoms such as wilt and necrosis of apical shoots were observed in 10 % of the cashew trees grown in a 100 plant-orchard in Bolama Island at Bijagós archipelago, Guinea-Bissau. Six symptomatic apical shoots from individual plants were collected for fungal isolation and identification. Tissue pieces (3 × 2 mm) from healthy to diseased margins were surface sterilized with 1 % sodium hypochlorite, washed twice with sterilized water, placed on potato dextrose agar (PDA, Difco® Laboratories) supplemented with potassium thiocyanate (50 µg/ml), and incubated at 24 ± 1 °C in the dark for 7 days. Four fungal colonies were isolated (67 %) and purified through hyphal tips removal, displaying rapid growth rate, and aerial mycelia that initially was white, turning later to dark greenish on PDA. Pycnidia produced on 1.5 % water agar and sterilized pine needles (± 25ºC; near-UV light) were solitary, covered by mycelium, obpyriform to ampulliform (152.5 ± 41.6 × 135.2 ± 30.8 µm, n = 30). Conidia were unicellular, hyaline, smooth, fusoid to ovoid, thin-walled, measuring 16.21 ± 1.52 × 5.84 ± 0.66 µm (n = 50, L/W 2.8). Such morphological features are characteristic of Neofusicoccum spp. (Phillips et al. 2013). For molecular identification, genomic DNA was extracted from a representative isolate GB160 and partial regions of ribosomal internal transcriber spacer (ITS) (ITS1/ITS4; White et al. 1990), translation elongation factor 1-α (EF1-α) (EF1-688F/EF1-1251R; Alves et al. 2008) and ß-tubulin (ß-tub) (Bt2a/Bt2b; Glass and Donaldson 1995) genes were amplified as previously described, respectively, with BSA (50 mg/ml). Amplicons were sequenced and deposited in GenBank (ITS, MN952993; EF1-α, MN952204; ß-tub, MN952208). BLAST analysis of ITS, EF1-α and ß-tub gene sequences showed 100 % identity with Neofusicoccum batangarum reference strain CBS124923 (FJ900608, FJ900654, FJ900635, respectively). Maximum-likelihood and Bayesian Inference analyses from the concatenated dataset placed GB160 isolate within the N. batangarum cluster (BS = 72 %; PP = 0.95). For pathogenicity assessment, 3-month-old cashew "Caju di Terra" plants (n = 8) grown in a greenhouse under controlled conditions were inoculated following a randomized block design as described by Lima et al. (2013). Briefly, 3 mm diam. stem tissue bark was removed and replaced with a 3 mm diameter PDA plug retrieved from the colony margin. Inoculation wound was covered with sterilized wet cotton and sealed with parafilm. Eight control plants were only treated with PDA plugs and the wound covered and sealed as described. After 15 days, all inoculated plants displayed similar symptoms to those observed in the field, and vascular lesions (10.8 ± 4.0 cm), whereas control plants remained symptomless. Koch's postulates were fulfilled by successful re-isolation of the pathogen from all inoculated stems and identification by morphology and gene sequencing. N. batangarum was identified associated with Anacardium spp. in Brazil (Netto et al. 2017) and recently reported as causing grapevine dieback in Brazil (Rêgo et al. 2020). To our knowledge, this is the first report of N. batangarum causing cashew dieback in Guinea-Bissau and West Africa. Occurrence of this disease may represent a significant impact for cashew production since this crop is the major agricultural commodity in Guinea-Bissau.

Integrating metabolomics and targeted gene expression to uncover potential biomarkers of fungal/oomycetes-associated disease susceptibility in grapevine.

Vitis vinifera, one of the most cultivated fruit crops, is susceptible to several diseases particularly caused by fungus and oomycete pathogens. In contrast, other Vitis species (American, Asian) display different degrees of tolerance/resistance to these pathogens, being widely used in breeding programs to introgress resistance traits in elite V. vinifera cultivars. Secondary metabolites are important players in plant defence responses. Therefore, the characterization of the metabolic profiles associated with disease resistance and susceptibility traits in grapevine is a promising approach to identify trait-related biomarkers. In this work, the leaf metabolic composition of eleven Vitis genotypes was analysed using an untargeted metabolomics approach. A total of 190 putative metabolites were found to discriminate resistant/partial resistant from susceptible genotypes. The biological relevance of discriminative compounds was assessed by pathway analysis. Several compounds were selected as promising biomarkers and the expression of genes coding for enzymes associated with their metabolic pathways was analysed. Reference genes for these grapevine genotypes were established for normalisation of candidate gene expression. The leucoanthocyanidin reductase 2 gene (LAR2) presented a significant increase of expression in susceptible genotypes, in accordance with catechin accumulation in this analysis group. Up to our knowledge this is the first time that metabolic constitutive biomarkers are proposed, opening new insights into plant selection on breeding programs.

Blanching impact on pigments, glucosinolates, and phenolics of dehydrated broccoli by-products.

Because of high water content, the valorisation of broccoli by-products requires dehydration that can preserve bioactive compounds. Blanching pre-treatment has been reported to improve the drying rate of broccoli. As a thermal treatment, it promotes also enzyme inactivation. Therefore, in this study, the impact of pre-dehydration blanching step, freeze-drying, air-drying at 40 °C, and microwave hydrodiffusion and gravity (MHG) dehydration on the levels of pigments, glucosinolates, and phenolics, was evaluated by UHPLC-DAD-ESI/MSn. When compared to freeze-drying, a technique known to preserve compounds, a pre-blanching step increased the extractability of both pigments and phenolics, while air-drying only retained 49% of the pigments and 70% of phenolics, both without affecting glucosinolates. However, when air-drying was preceded by blanching, less than 50% of compounds were retained. On the other hand, MHG dehydration increased the phenolics extractability by 26%, particularly that of kaempferol derivatives while also retaining the amount of the glucosinolate glucoraphanin, when compared to freeze-drying. Nevertheless, only 23% of indole glucosinolates were recovered and pigments were severely reduced, with lutein accounting only for 32% and only chlorophyll b was observed in trace amounts after MHG dehydration. Therefore, to valorise broccoli by-products as ingredients, different drying technologies may be used when targeting different composition richness: freeze-drying is suitable for pigments and glucosinolates, air-drying is suitable for glucosinolates, while MHG promotes the extractability of phenolic compounds.

Exploring glucosinolates diversity in Brassicaceae: a genomic and chemical assessment for deciphering abiotic stress tolerance.

Brassica is one of the most economically important genus of the Brassicaceae family, encompassing several key crops like Brassica napus (cabbage) and broccoli (Brassica oleraceae var. italica). This family is well known for their high content of characteristic secondary metabolites such as glucosinolates (GLS) compounds, recognize for their beneficial health properties and role in plants defense. In this work, we have looked through gene clusters involved in the biosynthesis of GLS, by combining genomic analysis with biochemical pathways and chemical diversity assessment. A total of 101 Brassicaceae genes involved in GLS biosynthesis were identified, using a multi-database approach. Through a UPGMA and PCA analysis on the 101 GLS genes recorded, revealed a separation between the genes mainly involved in GLS core structure synthesis and genes belonging to the CYP450s and MYBs gene families. After, a detailed phylogenetic analysis was conducted to better understand the disjunction of the aliphatic and indolic genes, by focusing on CYP79F1-F2 and CYP81F1-F4, respectively. Our results point to a recent diversification of the aliphatic CYP79F1 and F2 genes in Brassica crops, while for indolic genes an earliest diversification is observed for CYP81F1-F4 genes. Chemical diversity revealed that Brassica crops have distinct GLS chemo-profiles from other Brassicaceae genera; being highlighted the high contents of GLS found among the Diplotaxis species. Also, we have explored GLS-rich species as a new source of taxa with great agronomic potential, particularly in abiotic stress tolerance, namely Diplotaxis, the closest wild relatives of Brassica crops.

The leaf lipid composition of ectomycorrhizal oak plants shows a drought-tolerance signature.

Ectomycorrhizas have been reported to increase plant tolerance to drought. However, the mechanisms involved are not yet fully understood. Membranes are the first targets of degradation during drought, and growing evidences support a role for membrane lipids in plant tolerance and adaptation to drought. We have previously shown that improved tolerance of ectomycorrhizal oak plants to drought could be related to leaf membrane lipid metabolism, namely through an increased ability to sustain fatty acid content and composition, indicative of a higher membrane stability under stress. Here, we analysed in deeper detail the modulation of leaf lipid metabolism in oak plants mycorrhized with Pisolithus tinctorius and subjected to drought stress. Results show that mycorrhizal plants show patterns associated with water deficit tolerance, like a higher content of chloroplast lipids, whose levels are maintained upon drought stress. Likewise, mycorrhizal plants show increased levels of unsaturated fatty acids in the chloroplast phosphatidylglycerol lipid fraction. As a common response to drought, the digalactosyldiacyloglycerol/monogalactosyldiacyloglycerol ratio increased in the non-mycorrhizal plants, but not in the mycorrhizal plants, associated to smaller alterations in the expression of galactolipid metabolism genes, indicative of a higher drought tolerance. Under drought, inoculated plants showed increased expression of genes involved in neutral lipids biosynthesis, which could be related to an increased ability to tolerate drought stress. Overall, results from this study provide evidences of the involvement of lipid metabolism in the response of ectomycorrhizal plants to water deficit and point to an increased ability to maintain a stable chloroplast membrane functional integrity under stress.

Edible Leafy Vegetables from West Africa (Guinea-Bissau): Consumption, Trade and Food Potential.

Wild Edible Plants are common in the diet of rural communities of sub-Saharan Africa. In Guinea-Bissau, West Africa, wild plant resources are widely used in human diet, but very few studies have addressed them. The aim of this study is to reveal: (1) the wild and semi-cultivated leafy vegetables consumed in Guinea-Bissau; and (2) the nutritional composition of those plants traded at the largest country market in Bissau. Our results revealed that 24 native or naturalized species with edible leaves are currently consumed by Guinea-Bissau population. Five of them were found at the market: dried leaves of Adansonia digitata, Bombax costatum and Sesamum radiatum, and fresh leaves and shoots of Amaranthus hybridus and Hibiscus sabdariffa. The analysis of the nutritional properties revealed that leaves contain a significant amount of protein (10.1-21.0 g/100 g, dry basis), high values of macronutrients and micronutrients, as well as of phenolic compounds (13.1-40.3 mg GAE/g) and a considerable antioxidant capacity (DPPH 111.5-681.9 mg Eq Trolox). Although price and availability vary among the leafy vegetables analyzed, these traditional foods appear to be a good dietary component that can contribute to food security in Guinea-Bissau and in other West African countries, as these species are widely distributed in this region.