ABSTRACT Despite its interesting agricultural traits, Swingle citrumelo (Citrus paradisi Macfad. ‘Duncan’ × Poncirus trifoliata L. Raf.) rootstock exhibits severe incompatibility when grafted with ...Pera sweet orange Citrus sinensis (L.) Osb. scions, and this fact limits its use for propagation purposes. Nonetheless, the existence of certain Pera sweet orange clones that are compatible with Swingle citrumelo represents a substantial gain for the entire citrus industry. This study investigated the compatibility between seven Pera sweet orange clones and Swingle citrumelo rootstock, regarding the presence of a necrotic line at the graft union junction, the difference in diameter between rootstock and scion, the influence of clonal cleaning by micrografting, and the horticultural performance of the selected combinations. No trees died because of grafting incompatibility, even after twenty years (trial I), except for clone CV1, or after six years and nine months (trial II) from planting. The rootstock/scion trunk diameter ratio was not related to the presence of a necrotic line in the graft-union region. Additionally, these variables did not influence fruit yield and quality, and they were not affected by clonal cleaning or pre-immunisation of the old Pera sweet orange clones.
The use of temporary immersion systems (TIS) for plant micropropagation is an efficient technique for plant production, and we have applied it for the production of alstroemerias. This method ...involves the cultivation of explants such as rhizomes and axillary buds in a nutrient medium to stimulate shoot growth. TIS offer advantages such as accelerated multiplication processes, uniform production, and cost reduction. This process has shown promise in meeting the growing demand for alstroemeria plants in the market. This chapter describes a specific protocol for temporary immersion bioreactor micropropagation of the "Albatroz" cultivar, with the potential for large-scale automation.
The main difficulty for the cultivation and conservation of bromeliad species is the reduced number of propagules and slow growth of many of the species, resulting in a low propagation efficiency. ...Bromeliad plants are hardy and relatively easy to cultivate, with a high ornamental and ecological importance. Aiming at efficient micropropagation rates of V. hieroglyphica, a highly valued bromeliad, with very low propagation efficiency, a temporary immersion system was used and compared to semisolid and liquid static medium. Cultures obtained from in vitro germinated seeds were used as explants, maintaining their genetic diversity. Micropropagation with this simple temporary immersion system, composed of two autoclavable flasks, each with one opening for the attachment of 22 μm syringe filters, connected by a rubber stopper and an inner glass tube. In the bottom flask, an air valve is attached to the filter, which is subsequently connected to an aquarium pump and a timer and plugged to an outlet. This simple temporary immersion system showed improved micropropagation efficiency and is a method that can also be evaluated for other species.
A group of Gram-negative plant-associated diazotrophic bacteria belonging to the genus
Nitrospirillum
was investigated, including both previously characterized and newly isolated strains from diverse ...regions and biomes, predominantly in Brazil. Phylogenetic analysis of 16S rRNA and
recA
genes revealed the formation of a distinct clade consisting of thirteen strains, separate from the formally recognized species
N. amazonense
(the closest species) and
N. iridis
. Comprehensive taxonomic analyses using the whole genomes of four strains (BR 11140
T
= AM 18
T
= Y-2
T
= DSM 2788
T
= ATCC 35120
T
, BR 11142
T
= AM 14
T
= Y-1
T
= DSM 2787
T
= ATCC 35119
T
, BR 11145 = CBAmC, and BR 12005) supported the division of these strains into two species:
N. amazonense
(BR 11142
T
and BR 12005) and a newly proposed species (BR 11140
T
and BR 11145), distinct from
N. iridis
. The phylogenomic analysis further confirmed the presence of the new
Nitrospirillum
species. Additionally, MALDI-TOF MS analysis of whole-cell mass spectra provided further evidence for the differentiation of the proposed
Nitrospirillum
species, separate from
N. amazonense
. Analysis of chemotaxonomy markers (i.e., genes involved in fatty acid synthesis, metabolism and elongation, phospholipid synthesis, and quinone synthesis) revealed that the new species highlights high similarity and evolutionary convergence with other
Nitrospirillum
species. This new species exhibited nitrogen fixation ability
in vitro
, it has similar NifHDK protein phylogeny position with the closest species, lacked denitrification capability, but possessed the
nosZ
gene, enabling N
2
O reduction, distinguishing it from the closest species. Despite being isolated from diverse geographic regions, soil types, and ecological niches, no significant phenotypic or physiological differences were observed between the proposed new species and
N. amazonense
. Based on these findings, a new species,
Nitrospirillum viridazoti
sp. nov., was classified, with the strain BR 11140
T
(DSM 2788
T
, ATCC 35120
T
) designated as the type strain.
Atualmente o Brasil ocupa lugar de destaque entre os produtores de citros sendo o maior produtor de laranja doce do mundo. Entretanto, essa produção vem sendo gravemente afetada, por doenças como o ...huanglongbing (HLB), que tem causado perdas significativas para toda cadeia citrícola. O HLB está associado a bactérias Gram-negativas, restritas ao floema das plantas, denominadas Candidatus Liberibacter spp., que além de reduzir a produção de frutos, podem, em casos mais severos da doença, ocasionar a morte da planta. Uma importante estratégia para controle desta doença é a produção de plantas transgênicas expressando genes, especificamente no local de colonização do patógeno. O objetivo deste trabalho foi a obtenção de plantas transgênicas de Citrus sinensis cvs. \'Pera\' e \'Natal\', contendo o gene atacina A (attA) que codifica um peptídeo antibacteriano, dirigido por promotores preferencialmente expressos no floema: AtSuc2 (transportador de sacarose) ou AtPP2 (proteína de floema 2), clonados de Arabidopsis thaliana, ou CsPP2 (proteína de floema 2) clonado de Citrus sinensis. A identificação de 65 plantas transgênicas foi realizada, por meio da análise de PCR. Foi verificado um número menor de eventos transgênicos, utilizando-se a construção gênica pCAtSuc2/attA em relação ao número de eventos transgênicos obtidos com as construções gênicas pCCsPP2/attA e pCAtPP2/attA. Plantas identificadas como transgênicas pela análise de PCR, foram aclimatizadas e transferidas para casa-devegetação certificada para o cultivo de plantas transgênicas. Análises de Southern blot foram realizadas em plantas aclimatizadas que apresentaram desenvolvimento suficiente, confirmando-se a integração do gene attA. A expressão do gene attA foi confirmada pela análise de RT-qPCR. As plantas transgênicas obtidas neste trabalho, contendo o gene attA dirigido por promotores preferencialmente expressos no floema, serão avaliadas em uma etapa futura para resistência a Candidatus Liberibacter spp.
Currently, Brazil is a major citrus producer and the world\'s largest producer of sweet oranges. However, diseases, such as huanglongbing (HLB) have seriously affected this production, causing significant losses in citrus production chain. HLB is associated with Gram-negative bacterias, restricted to the phloem of plants, called Candidatus Liberibacter spp., which besides reducing fruit production, can lead to plant death. An important strategy to control this disease is the production of transgenic plants expressing genes, specifically at the region of pathogen colonization. The aim of this study was to obtain transgenic plants of Citrus sinensis cv. \'Natal\' and \'Pera\', containing the gene attacin A (attA) that encodes an antibacterial peptide, driven by preferentially expressed in phloem promoters: AtSuc2 (sucrose transporter) or AtPP2 (phloem protein 2), cloned from Arabidopsis thaliana, or CsPP2 (phloem protein 2) cloned from Citrus sinensis. The identification of 65 transgenic plants was performed by PCR analysis. A lower number of transgenic events were verified using the gene construct pCAtSuc2/attA in relation events obtained with the gene constructs pCCsPP2/attA and pCAtPP2/attA. Plants identified as transgenic by PCR analysis were acclimatized and transferred to a greenhouse certified for growing transgenic plants. The Southern blot analyses were performed in acclimatized plants, which had sufficiently developed, confirming the integration of attA gene. The expression of attA gene was confirmed by RT-qPCR analysis. The transgenic plants obtained in this work, containing the gene attA directed by preferentially expressed in phloem promoters, will be further evaluated for resistance to Candidatus Liberibacter spp.
Benign prostatic hyperplasia (BPH) is characterised by increases in prostate volume and contraction. Downregulation of the nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) signalling pathway ...contributes to prostate dysfunctions. Previous studies in cancer cells or vessels have shown that the epigenetic mechanisms control the gene and protein expression of the enzymes involved in the production of NO and cGMP. This study is aimed to evaluate the effect of a 2-week treatment of 5-azacytidine (5-AZA), a DNA-methyltransferase inhibitor, in the prostate function of mice fed with a high-fat diet. Functional, histological, biochemical and molecular assays were carried out. Obese mice presented greater prostate weight, α-actin expression and contractile response induced by the α-1adrenoceptors agonist. The relaxation induced by the NO-donor and the protein expression of endothelial nitric oxide synthase (eNOS) and soluble guanylate cyclase (sGC) were significantly decreased in the prostate of obese mice. The treatment with 5-AZA reverted the higher expression of α-actin, reduced the hypercontractility state of the prostate and increased the expression of eNOS and sGC and intraprostatic levels of cGMP. When prostates from obese mice treated with 5-AZA were incubated in vitro with inhibitors of the NOS or sGC, the inhibitory effect of 5-AZA was reverted, therefore, showing the involvement of NO and cGMP. In conclusion, our study paves the way to develop or repurpose therapies that recover the expression of eNOS and sGC and, hence, to improve prostate function in BPH.
Introduction
Clinical trials on tyrosine kinase inhibitors (TKI) treatment have shown an improvement in overall and progression‐free survival in patients with advanced differentiated thyroid cancer. ...However, it is necessary to evaluate these studies to assess methodological biases and inconsistencies that may impact the effects.
Objective
To map and assess the methodological quality of randomized clinical trials (RCTs) regarding randomization, allocation concealment, blinding, and selective reporting bias.
Methods
RCTs assessing the efficacy and safety of TKI for the treatment of advanced differentiated thyroid cancer were included. The search was performed in the MEDLINE database. The included RCTs were assessed for the adequacy of the methodological steps, as recommended by the Cochrane Risk of Bias tool.
Results
Nine studies were analyzed, of which 77.7% were classified as low risk of bias regarding selective reporting and 33.3% as high risk of reporting bias. The mean time between protocol registration and study publication was approximately 5.11 years. Moreover, 66.7% were classified as low risk of bias for randomization and allocation concealment, and 33.3% did not specify the randomization process and allocation concealment in a way that would allow the identification of occurrences of bias. Concerning blinding of participants and outcome assessors, 77.8% of the RCTs reported adequate blinding and were classified as having a low risk of bias, 11.1% had a high risk of bias, and 11.1% had insufficient information and were classified as having unclear risk of bias. Regarding the blinding of the outcome assessors, 33.3% did the blinding correctly, 11.1% did not blind, and 55.6% did not provide enough information.
Conclusion
Overall, the assessed RCTs were predominantly at low risk of bias. The critical evaluation of these studies is essential to have confidence in the treatment estimated effect that will support clinical decision‐making and provide information to preclude future clinical study flaws.
Common bean (Phaseolus vulgaris L.) seeds are an economical protein source rich in bioactive compounds with antioxidant activity. In this work, we evaluated processed seeds from two common bean ...cultivars regarding protein profile and antioxidant capacity before and after enzymatic digestion in vitro. We provided protein maps by two‐dimensional electrophoresis (2‐DE) of seeds germinated during 36 h and 72 h. The principal component analysis (PCA) showed that protein abundance had more variation on germination times than the cultivar ones. Seeds germinated for 36 h showed greater antioxidant capacity compared to germination for 72 h and cooking, before and after enzymatic digestion. In protein isolates, cooking associated with digestion provided better antioxidant capacity. Germination time influences protein expression as well as the antioxidant capacity of common beans. Furthermore, in vitro digestion increases the antioxidant capacity of cooked bean protein isolates.
Protein profile and antioxidant capacity of processed seeds from two common bean cultivars.
ACE2 and TMPRSS2 are key players on SARS-CoV-2 entry into host cells. However, it is still unclear whether expression levels of these factors could reflect disease severity. Here, a case-control ...study was conducted with 213 SARS-CoV-2 positive individuals where cases were defined as COVID-19 patients with respiratory distress requiring oxygen support (N = 38) and controls were those with mild to moderate symptoms of the disease who did not need oxygen therapy along the entire clinical course (N = 175). ACE2 and TMPRSS2 mRNA levels were evaluated in nasopharyngeal swab samples by RT-qPCR and logistic regression analyzes were applied to estimate associations with respiratory outcomes. ACE2 and TMPRSS2 levels positively correlated with age, which was also strongly associated with respiratory distress. Increased nasopharyngeal ACE2 levels showed a protective effect against this outcome (
OR = 0.30; 95% CI 0.09-0.91), while TMPRSS2/ACE2 ratio was associated with risk (
OR = 4.28; 95% CI 1.36-13.48). On stepwise regression, TMPRSS2/ACE2 ratio outperformed ACE2 to model COVID-19 severity. When nasopharyngeal swabs were compared to bronchoalveolar lavages in an independent cohort of COVID-19 patients under mechanical ventilation, similar expression levels of these genes were observed. These data suggest nasopharyngeal TMPRSS2/ACE2 as a promising candidate for further prediction models on COVID-19.
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