Fine-Tuning of Arabidopsis thaliana Response to Endophytic Colonization by Gluconacetobacter diazotrophicus PAL5 Revealed by Transcriptomic Analysis.

Gluconacetobacter diazotrophicus is a diazotrophic endophytic bacterium that promotes the growth and development of several plant species. However, the molecular mechanisms activated during plant response to this bacterium remain unclear. Here, we used the RNA-seq approach to understand better the effect of G. diazotrophicus PAL5 on the transcriptome of shoot and root tissues of Arabidopsis thaliana. G. diazotrophicus colonized A. thaliana roots and promoted growth, increasing leaf area and biomass. The transcriptomic analysis revealed several differentially expressed genes (DEGs) between inoculated and non-inoculated plants in the shoot and root tissues. A higher number of DEGs were up-regulated in roots compared to shoots. Genes up-regulated in both shoot and root tissues were associated with nitrogen metabolism, production of glucosinolates and flavonoids, receptor kinases, and transcription factors. In contrast, the main groups of down-regulated genes were associated with pathogenesis-related proteins and heat-shock proteins in both shoot and root tissues. Genes encoding enzymes involved in cell wall biogenesis and modification were down-regulated in shoots and up-regulated in roots. In contrast, genes associated with ROS detoxification were up-regulated in shoots and down-regulated in roots. These results highlight the fine-tuning of the transcriptional regulation of A. thaliana in response to colonization by G. diazotrophicus PAL5.

Predictive value of ultrasound doppler parameters in neoadjuvant chemotherapy response of breast cancer: Prospective comparison with magnetic resonance and mammography.

BACKGROUND: Neoadjuvant chemotherapy (NACT) is a treatment option for breast cancer patients that allows for the assessment of tumor response during treatment. This information can be used to adjust treatment and improve outcomes. However, the optimal imaging modalities and parameters for assessing tumor response to NACT are not well established. METHODS: This study included 173 breast cancer patients who underwent NACT. Patients were imaged with ultrasound (US), mammography (MMG), and magnetic resonance imaging (MRI) at baseline, after two cycles of NACT, and before breast surgery. US parameters included lesion morphology, Doppler variables, and elastography measurements. MMG and MRI were evaluated for the presence of nodules and tumor dimensions. The pathological response to NACT was determined using the residual cancer burden (RCB) classification. RESULTS: The US parameter with the highest power for predicting pathological complete response (pCR) was shear wave elastography (SWE) maximum speed inside the tumor at baseline. For nonluminal tumors, the end diastolic velocity measured by US after two cycles of NACT showed the highest predictive value for pCR. Similarly, SWE maximum speed after two cycles of NACT had the highest discriminating power for predicting RCB-III in luminal tumors, while the same parameter measured at baseline was most predictive for nonluminal tumors. CONCLUSIONS: This study provides evidence that mid-treatment Doppler US and other imaging modalities can be used to predict the response to NACT in breast cancer patients. Functional parameters, such as blood flow velocities and SWE measurements, demonstrated superior predictive value for pCR, while morphological parameters had limited value. These findings have implications for personalized treatment strategies and may contribute to improved outcomes in the management of breast cancer.

Coding, Decoding and Retrieving a Message Using DNA: An Experience from a Brazilian Center Research on DNA Data Storage.

DNA data storage based on synthetic oligonucleotides is a major attraction due to the possibility of storage over long periods. Nowadays, the quantity of data generated has been growing exponentially, and the storage capacity needs to keep pace with the growth caused by new technologies and globalization. Since DNA can hold a large amount of information with a high density and remains stable for hundreds of years, this technology offers a solution for current long-term data centers by reducing energy consumption and physical storage space. Currently, research institutes, technology companies, and universities are making significant efforts to meet the growing need for data storage. DNA data storage is a promising field, especially with the advancement of sequencing techniques and equipment, which now make it possible to read genomes (i.e., to retrieve the information) and process this data easily. To overcome the challenges associated with developing new technologies for DNA data storage, a message encoding and decoding exercise was conducted at a Brazilian research center. The exercise performed consisted of synthesizing oligonucleotides by the phosphoramidite route. An encoded message, using a coding scheme that adheres to DNA sequence constraints, was synthesized. After synthesis, the oligonucleotide was sequenced and decoded, and the information was fully recovered.

Silencing ScGUX2 reduces xylan glucuronidation and improves biomass saccharification in sugarcane.

There is an increasing need for renewable energy sources to replace part of our fossil fuel-based economy and reduce greenhouse gas emission. Sugarcane bagasse is a prominent feedstock to produce cellulosic bioethanol, but strategies are still needed to improve the cost-effective exploitation of this potential energy source. In model plants, it has been shown that GUX genes are involved in cell wall hemicellulose decoration, adding glucuronic acid substitutions on the xylan backbone. Mutation of GUX genes increases enzyme access to cell wall polysaccharides, reducing biomass recalcitrance in Arabidopsis thaliana. Here, we characterized the sugarcane GUX genes and silenced GUX2 in commercial hybrid sugarcane. The transgenic lines had no penalty in development under greenhouse conditions. The sugarcane GUX1 and GUX2 enzymes generated different patterns of xylan glucuronidation, suggesting they may differently influence the molecular interaction of xylan with cellulose and lignin. Studies using biomass without chemical or steam pretreatment showed that the cell wall polysaccharides, particularly xylan, were less recalcitrant in sugarcane with GUX2 silenced than in WT plants. Our findings suggest that manipulation of GUX in sugarcane can reduce the costs of second-generation ethanol production and enhance the contribution of biofuels to lowering the emission of greenhouse gases.

Pogonias courbina sperm characteristcs in its first reproductive season.

Southern black drum (Pogonias courbina) is a species distributed along the western Atlantic Ocean, and it is the largest Sciaenidae observed in the coast of Rio Grande do Sul state, Brazil. However, it is listed as a vulnerable species at The IUCN Red List of Threatened Species™, and their fishing is prohibited. The objective of this study was to determine the sperm characteristics of P. courbina. Sperm samples of five young males (two-year-old fish) were collected through abdominal pressure. The sperm kinetics parameters were sperm motility (MOT) 10.7 ± 5.6%, curvilinear velocity (VCL) 120.07 ± 16.16 mm s ± 1, average path velocity (VAP) 75.64 ± 23.78 mm s ± 1, straight-line velocity (VSL) 62.49 ± 15.83 mm s ± 1, straightness (STR) 83.9 ± 5.3%, wobble (WOB) 61.9 ± 12.7%, beat cross frequency (BCF) 42.981 ± 4.627 Hz and progression (PRG) 1,805.4 ± 564.5 µm. The proportion of normal spermatozoa was 35.6 ± 6.1%. About the abnormalities observed, 22.7% occurred in the tail (short tail = 0.6 ± 0.5%, distally curled tail = 2.4 ± 1.6%, strongly curled tail = 1.9 ± 1.3%, broken tail = 7.9 ± 5.1%, folded tail = 5.5 ± 0.8%, loose tail = 4.4 ± 1.9%); 14.2% occurred in the head (degenerate head = 4.2 ± 1.6%, microcephaly = 1.8 ± 2.5%, loose head = 8.2 ± 2.1%) and 27.5% of the spermatozoa showed cytoplasmatic gouts (proximal gout = 20.0 ± 8.4%, distal gout = 7.5 ± 2.8%). Besides that, a correlation analysis was performed between sperm morphology and kinetics parameters, and the spermatozoa were measured for the morphometric parameters. There was a positive correlation between BCF and normal spermatozoa (r =  0.9269). A negative correlation occurred between BCF and loose head (r =  -0.9047); WOB and strongly curled tail (r =  -0.8911); and PROG and strongly curled tail (r =  -0.9191). The morphometric measures found for the head were length of 2.50 ± 0.21 µm and width of 2.12 ± 0.22 µm, and for the tail it was length of 37.97 ± 2.01 µm. It was possible to verify that the animals have sperm characteristics that indicate reproductive aptitude, but an abnormal behavior on sperm activation and high presence of the cytoplasmic gout abnormality indicates that the animals are not fully mature in their first reproductive season. This work contributes to a better understanding of the P. courbina spermatic parameters, what can be allies to recovery this species population in nature and promote its production in fish farms.

Bio-nanocomposites films based on unmodified and modified thermoplastic starch reinforced with chemically modified nanoclays.

The use of polymers capable of being degraded by the action of microorganisms and/or enzymes without causing harmful effects is a strategy in waste management and environmental care. In this work, bio-nanocomposites based on thermoplastic starch (TPS) were synthesized by reactive extrusion using a twin-screw extruder. Two strategies were evaluated to reduce the disadvantages of TPS for packaging applications. First, starch was chemically modified producing the reaction of native starch with chemical reagents that introduce new functional groups to reduce the water adsorption. And two, nano-fillers were incorporated into TPS in order to enhance the mechanical and barrier properties, driving to materials with improved performance/cost ratio. The synergistic strategies of chemical modification and incorporation of modified nanoclays were also effective to reduce the dependence of properties of TPS with the environment humidity and the evolution thereof over time, which influences the performance during the service life of the product. Supplementary Information: The online version contains supplementary material available at 10.1007/s10853-023-08354-1.

Do final rehabilitation volumes influence function after anterior cross ligament reconstruction? A pilot study / Volumes finais de reabilitação influenciam função após reconstrução de ligamento cruzado anterior? Um estudo piloto

Abstract Introduction Rupture of the anterior cruciate ligament (ACL) is one of the frequent traumatic injuries of the knee joint complex, and the isokinetic dynamometer is essential to assess and measure its joint function. Objective To analyze the volume of physical therapy treatment using the same protocol with different duration in patients undergoing ACL reconstruction. Methods This is a prospective observational cohort study. The sample consisted of 13 adult male individuals who underwent ACL reconstruction surgery. Participants were divided into two groups: accelerated treatment (AC) and non-accelerated treatment (NAC). Participants underwent a standardized protocol of post-reconstruction physio-therapy, starting in the immediate postoperative period. The AC group performed the treatment three times a week, with a duration of 4 hours each session for 6 months, while the NAC performed twice a week with duration of two hours each session for eight months. Results The groups showed the same behavior during the post-reconstruction treatment periods, showing that regardless of time, the results were beneficial at the end of the treatment. However, in the accelerated protocol, the injured limb showed a significant difference for extender peak torque (PTEXT) in the preoperative and 4-month postoperative period (230.5 vs 182.6), 4-month postoperative period and final (182.6 vs 242.1) in the AC group, while in the NAC group there was no significant difference between times in this same limb. Conclusion Greater volumes of weekly training characterized better results, showing that rehabilitation time is not a predictor of discharge, but rehabilitation time obtains good results for the variables.

Resumo Introdução A ruptura do ligamento cruzado anterior (LCA) é uma das lesões traumáticas frequentes do complexo articular do joelho, sendo o dinamômetro isocinético fundamental para avaliar e mensurar sua função articular. Objetivo Analisar o volume de tratamento fisioterapêutico utilizando o mesmo protocolo com diferentes durações em pacientes submetidos à reconstrução do LCA. Métodos Trata-se de um estudo de coorte observacional prospectivo. A amostra foi composta por 13 indivíduos adultos do sexo masculino submetidos à cirurgia de reconstrução do LCA. Os participantes foram divididos em dois grupos: tratamento acelerado (AC) e tratamento não acelerado (NAC). Os participantes foram submetidos a um protocolo padronizado de fisioterapia pós-reconstrução, com início no pós-operatório imediato. O grupo AC realizou o trata-mento três vezes por semana, com duração de 4 horas cada sessão, durante 6 meses, enquanto o grupo NAC realizou duas vezes por semana, com duração de duas horas cada sessão, durante oito meses. Resultados Os grupos apresentaram o mesmo comportamento durante os períodos de tratamento pós-reconstrução, mostrando que independente do tempo, os resultados foram benéficos ao final do tratamento. No protocolo acelerado, porém, o membro lesionado apresentou diferença significativa para pico de torque extensor (PTEXT) no pré-operatório e pós-operatório de 4 meses (230,5 vs 182,6), pós-operatório de 4 meses e final (182,6 vs 242,1) no grupo AC, enquanto no NAC não houve diferença significativa entre os tempos neste mesmo membro. Conclusão Maiores volumes de treinamento semanal caracterizaram melhores resultados, mostrando que o tempo de reabilitação não é preditor de alta, mas o tempo de reabilitação obtém bons resultados para as variáveis.

Effects of Lidocaine Injection at Acupuncture Points on Perioperative Analgesia in Cats Undergoing Ovariohysterectomy.

Background: Pharmacopuncture is an acupuncture-related technique that has been used to amplify the therapeutic effects of different medications. Objectives: To investigate the analgesic efficacy of a lidocaine injection at acupoints in cats undergoing ovariohysterectomy. Methods: Thirty cats were randomly distributed into two groups (n = 15, per group). The experimental group received a bilateral administration of lidocaine at the following acupoints: Stomach 36 (ST-36) and Spleen 6 (SP-6) (Lido group). The control group did not receive lidocaine (Control group). All cats were sedated with dexmedetomidine and anesthesia was induced with propofol and maintained with isoflurane. Intraoperatively, fentanyl was given to control cardiovascular responses to surgical stimulation. Postoperative pain was assessed at various time points, up to 24 hours after extubation, using the UNESP-Botucatu multidimensional composite pain scale (MCPS) and Glasgow feline composite measure pain scale (CMPS-Feline). Sedation scores were measured at the same time points. Morphine/meloxicam was administered as rescue analgesia. Data were analyzed using t-tests, Fisher´s exact test, the Mann-Whitney test, and the Friedman test (p < 0.05). Results: Intraoperatively, more cats in the Control group required analgesic supplementation than those in the Lido group, but the difference was not significant (p = 0.65). Postoperative pain, sedation scores, and analgesic requirements did not differ between groups. Rescue analgesia was given to 67% (10/15) of the cats in each group. Conclusion: The administration of lidocaine at ST-36 and SP-6 acupuncture points did not provide significant perioperative analgesic benefits in healthy cats undergoing ovariohysterectomy.

Molecular and Computational Strategies to Increase the Efficiency of CRISPR-Based Techniques.

The prokaryote-derived Clustered Regularly Interspaced Palindromic Repeats (CRISPR)/Cas mediated gene editing tools have revolutionized our ability to precisely manipulate specific genome sequences in plants and animals. The simplicity, precision, affordability, and robustness of this technology have allowed a myriad of genomes from a diverse group of plant species to be successfully edited. Even though CRISPR/Cas, base editing, and prime editing technologies have been rapidly adopted and implemented in plants, their editing efficiency rate and specificity varies greatly. In this review, we provide a critical overview of the recent advances in CRISPR/Cas9-derived technologies and their implications on enhancing editing efficiency. We highlight the major efforts of engineering Cas9, Cas12a, Cas12b, and Cas12f proteins aiming to improve their efficiencies. We also provide a perspective on the global future of agriculturally based products using DNA-free CRISPR/Cas techniques. The improvement of CRISPR-based technologies efficiency will enable the implementation of genome editing tools in a variety of crop plants, as well as accelerate progress in basic research and molecular breeding.

Reduction of ethylene biosynthesis in sugarcane induces growth and investment in the non-enzymatic antioxidant apparatus.

KEY MESSAGE: Lower ethylene production in sugarcane results in plants with higher stature, expression of growth-promoting genes, higher photosynthetic rate, and increased antioxidant compounds. The hormone ethylene is involved in critical processes in sugarcane, such as the growth and accumulation of sucrose. The lack of mutants for ethylene biosynthesis or signaling genes makes it difficult to understand the role of this phytohormone throughout sugarcane development. This study aimed to evaluate the physiology and development of sugarcane plants with low ethylene production. To achieve this goal, we used RNA interference to silence three genes, ScACS1, ScACS2, and ScACS3, encoding 1-aminocyclopropane-1-carboxylic acid synthases (ACS), responsible for a limiting step of the ethylene biosynthesis pathway. Sugarcane plants with reduced ethylene levels presented increased growth, faster germination of lateral gems, and activation of non-enzymatic antioxidant mechanisms. We observed an augmentation in the expression of ScACO5, which encodes the final enzyme regulating ethylene biosynthesis, and ScERF1, encoding a transcription factor, linked to the ethylene response. The increase in plant height was correlated with higher expression of ScPIF3, ScPIF4, and ScPIF5, which encode for transcription factors related to growth induction. Interestingly, there was also an increase in the expression of the ScGAI gene, which encodes a DELLA protein, a growth repressor. The final content of sucrose in the stems was not affected by the low levels of ethylene, although the rate of CO2 assimilation was reduced. This study reports for the first time the impacts of low endogenous production of ethylene in sugarcane and provides helpful insights on the molecular mechanisms behind ethylene responses.

Amino Acid and Carbohydrate Metabolism Are Coordinated to Maintain Energetic Balance during Drought in Sugarcane.

The ability to expand crop plantations without irrigation is a major goal to increase agriculture sustainability. To achieve this end, we need to understand the mechanisms that govern plant growth responses under drought conditions. In this study, we combined physiological, transcriptomic, and genomic data to provide a comprehensive picture of drought and recovery responses in the leaves and roots of sugarcane. Transcriptomic profiling using oligoarrays and RNA-seq identified 2898 (out of 21,902) and 46,062 (out of 373,869) transcripts as differentially expressed, respectively. Co-expression analysis revealed modules enriched in photosynthesis, small molecule metabolism, alpha-amino acid metabolism, trehalose biosynthesis, serine family amino acid metabolism, and carbohydrate transport. Together, our findings reveal that carbohydrate metabolism is coordinated with the degradation of amino acids to provide carbon skeletons to the tricarboxylic acid cycle. This coordination may help to maintain energetic balance during drought stress adaptation, facilitating recovery after the stress is alleviated. Our results shed light on candidate regulatory elements and pave the way to biotechnology strategies towards the development of drought-tolerant sugarcane plants.

Plant 3' Regulatory Regions From mRNA-Encoding Genes and Their Uses to Modulate Expression.

Molecular biotechnology has made it possible to explore the potential of plants for different purposes. The 3' regulatory regions have a great diversity of cis-regulatory elements directly involved in polyadenylation, stability, transport and mRNA translation, essential to achieve the desired levels of gene expression. A complex interaction between the cleavage and polyadenylation molecular complex and cis-elements determine the polyadenylation site, which may result in the choice of non-canonical sites, resulting in alternative polyadenylation events, involved in the regulation of more than 80% of the genes expressed in plants. In addition, after transcription, a wide array of RNA-binding proteins interacts with cis-acting elements located mainly in the 3' untranslated region, determining the fate of mRNAs in eukaryotic cells. Although a small number of 3' regulatory regions have been identified and validated so far, many studies have shown that plant 3' regulatory regions have a higher potential to regulate gene expression in plants compared to widely used 3' regulatory regions, such as NOS and OCS from Agrobacterium tumefaciens and 35S from cauliflower mosaic virus. In this review, we discuss the role of 3' regulatory regions in gene expression, and the superior potential that plant 3' regulatory regions have compared to NOS, OCS and 35S 3' regulatory regions.

Bringing to light the molecular evolution of GUX genes in plants.

Hemicellulose and cellulose are essential polysaccharides for plant development and major components of cell wall. They are also an important energy source for the production of ethanol from plant biomass, but their conversion to fermentable sugars is hindered by the complex structure of cell walls. The glucuronic acid substitution of xylan (GUX) enzymes attach glucuronic acid to xylan, a major component of hemicellulose, decreasing the efficiency of enzymes used for ethanol production. Since loss-of-function gux mutants of Arabidopsis thaliana enhance enzyme accessibility and cell wall digestion without adverse phenotypes, GUX genes are potential targets for genetically improving energy crops. However, comprehensive identification of GUX in important species and their evolutionary history are largely lacking. Here, we identified putative GUX proteins using hidden Markov model searches with the GT8 domain and a GUX-specific motif, and inferred the phylogenetic relationship of 18 species with Maximum likelihood and Bayesian approaches. Each species presented a variable number of GUX, and their evolution can be explained by a mixture of divergent, concerted and birth-and-death evolutionary models. This is the first broad insight into the evolution of GUX gene family in plants and will potentially guide genetic and functional studies in species used for biofuel production.

O trabalho do enfermeiro no atendimento às gestantes: ações básicas, problemas comuns e a sistematização da assistência na consulta pré-natal / The work of nurses in caring for pregnant women: Basic actions, common problems and the systematization of care in prenatal consultation

A Atenção Primária à Saúde pode ser considerada como porta de acesso ao SUS, acolhendo diversos tipos de usuários com estado de saúde alterado ou estável. Sabe-se que, em relação ao atendimento materno, a maioria pode ser resolvida em unidades básicas de saúde e, portanto, os profissionais devem estar capacitados para prestar este tipo de atendimento. O enfermeiro deve ter conhecimentos no campo da saúde da mulher para o cuidado integral nos serviços materno-infantil, especialmente na assistência às gestantes. Este estudo teve como objetivo geral analisar o trabalho do enfermeiro no atendimento às gestantes na Atenção Primária à Saúde. Trata-se de estudo do tipo exploratório com abordagem qualitativa dos dados. O cenário de estudo foi constituído por sete unidades básicas de saúde de um município do interior do estado de São Paulo, e a amostra selecionada foram sete enfermeiros. Para a coleta de dados, foram utilizadas entrevistas semiestruturadas, no período de outubro de 2018 a fevereiro de 2019. A análise temática indutiva foi executada para interpretação dos dados. Este estudo foi aprovado pelo Comitê de Ética em Pesquisa da Instituição Proponente, e identificaram-se ações essenciais para o atendimento à gestante, como: acolhimento, consulta pré-natal, orientação nutricional e visitas domiciliares. Ademais, os discursos revelaram pouco conhecimento dos participantes sobre um método sistematizado para realizar o atendimento, como a Classificação Internacional das Práticas de Enfermagem (CIPE). Os dados permitiram identificar problemas comuns no atendimento às gestantes, tais como: dor aguda, edema, ansiedade, náuseas, além da presença de Hipertensão Arterial Sistêmica. Para tais problemas/diagnósticos, foram verbalizadas algumas intervenções. Como produto da dissertação foi elaborado um folheto explicativo com a finalidade de sistematizar a consulta de pré-natal, apresentando ações básicas, principais diagnósticos e intervenções de enfermagem. Este estudo deve provocar a reflexão dos gestores quanto à elaboração de estratégias que visam aprimorar saberes específicos nestes profissionais para um atendimento seguro e de qualidade às gestantes. Ademais, pode auxiliar centros formadores na implementação de metodologias inovadoras para a formação dos futuros enfermeiros para atuar nesta área.

Primary Health Care can be considered as a gateway to SUS, welcoming various types of users with altered or stable health status. It is known that in relation to maternal care, most can be resolved in basic health units and, therefore, professionals should be able to provide this type of care. The nurse must have knowledge in the field of women's health for comprehensive care in maternal and child services, especially in the care of pregnant women. This study aimed to analyze the work of nurses in the care of pregnant women in Primary Health Care. It is an exploratory study with a qualitative approach to data. The study scenario consisted of seven basic health units of a municipality in the interior of the state of São Paulo and the sample selected were seven nurses. For data collection, semi-structured interviews were used from October 2018 to February 2019. Inductive thematic analysis was used for data interpretation. Study approved by the Research Ethics Committee of the Proponent Institution. Essential actions for the care of pregnant women were identified as: reception, prenatal consultation, nutritional counseling and home visits. In addition, the speeches revealed little knowledge of participants about a systematized method to perform care such as the International Classification of Nursing Practices (ICNP). The data allowed us to identify common problems in in the care of pregnant women, such as acute pain, edema, anxiety, nausea and the presence of systemic arterial hypertension. For such problems / diagnoses some interventions were verbalized. As a result of the dissertation, an explanatory leaflet was elaborated to systematize the prenatal consultation, presenting basic actions, main diagnoses and nursing interventions. This study should provoke the reflection of the managers about the elaboration of strategies that aim to improve specific knowledge in these professionals for a safe and quality attendance to the pregnant women. It can also assist training centers in the implementation of innovative methodologies for the training of future nurses to work in this area.

The C-Terminal Domains SnRK2 Box and ABA Box Have a Role in Sugarcane SnRK2s Auto-Activation and Activity.

Resistance to drought stress is fundamental to plant survival and development. Abscisic acid (ABA) is one of the major hormones involved in different types of abiotic and biotic stress responses. ABA intracellular signaling has been extensively explored in Arabidopsis thaliana and occurs via a phosphorylation cascade mediated by three related protein kinases, denominated SnRK2s (SNF1-related protein kinases). However, the role of ABA signaling and the biochemistry of SnRK2 in crop plants remains underexplored. Considering the importance of the ABA hormone in abiotic stress tolerance, here we investigated the regulatory mechanism of sugarcane SnRK2s-known as stress/ABA-activated protein kinases (SAPKs). The crystal structure of ScSAPK10 revealed the characteristic SnRK2 family architecture, in which the regulatory SnRK2 box interacts with the kinase domain αC helix. To study sugarcane SnRK2 regulation, we produced a series of mutants for the protein regulatory domains SnRK2 box and ABA box. Mutations in ScSAPK8 SnRK2 box aimed at perturbing its interaction with the protein kinase domain reduced protein kinase activity in vitro. On the other hand, mutations to ScSAPK ABA box did not impact protein kinase activity but did alter the protein autophosphorylation pattern. Taken together, our results demonstrate that both SnRK2 and ABA boxes might play a role in sugarcane SnRK2 function.

Perinodal fibrosis developed after ultrasonography-guided core-needle biopsy of a contrast-enhanced ultrasound-detected sentinel axillary node interferes with subsequent surgical sentinel node dissection.

OBJECTIVE: To evaluate perinodal fibrosis after 14-gauge staging core-needle biopsy (CNB) of the axillary sentinel lymph node (SLN) identified using contrast-enhanced ultrasonography (CEUS) and its interference with subsequent surgical SLN dissection in breast cancer patients. METHODS: Frequencies or means of main clinical, sonographic, pathological, and surgical characteristics were calculated. We also compared patient groups with and without perinodal pathological fibrosis. RESULTS: Forty-eight patients who underwent CEUS + CNB and axillary surgery were eligible for this cross-sectional study. Axillary surgical specimens showed perinodal fibrosis in 9/48 (18.7%) patients. Interference with SLN dissection was reported in 4/48 (8.3%) patients (two hematomas, three abnormal palpation findings, and four difficult dissections). The overall surgical detection rate of SLN was 43/48 (89.6%). In the majority of cases, perinodal fibrosis was described as moderate (4/9 [44.4%]) or severe (4/9 [44.4%]). The mean time elapsed between CEUS + CNB and axillary dissection was shorter in patients with perinodal fibrosis (P = .04). Interference with SLN dissection was only reported in patients with perinodal fibrosis (P < .001). Surgical SLN detection was successful in all nine cases in which perinodal pathological fibrosis or interference with SLN dissection was reported. CONCLUSION: Perinodal fibrosis may impair the surgical SLN dissection in early stage breast cancer patients who were staged using CEUS + CNB using a14-gauge needle.

Overexpression of an evolutionarily conserved drought-responsive sugarcane gene enhances salinity and drought resilience.

BACKGROUND AND AIMS: Improving drought adaptation is more pressing for crops such as sugarcane, rice, wheat and maize, given the high dependence of these crops on irrigation. One option for enhancing adaptation to water limitation in plants is by transgenic approaches. An increasing number of genes that are associated with mechanisms used by plants to cope with water scarcity have been discovered. Genes encoding proteins with unknown functions comprise a relevant fraction of the genes that are modulated by drought. We characterized a gene in response to environmental stresses to gain insight into the unknown fraction of the sugarcane genome. Scdr2 (Sugarcane drought-responsive 2) encodes a small protein and shares highly conserved sequences within monocots, dicots, algae and fungi. METHODS: Plants overexpressing the Scdr2 sugarcane gene were examined in response to salinity and drought. Measurements of the gas exchange parameters, germination rate, water content, dry mass and oxidative damage were performed. Seeds as well as juvenile plants were used to explore the resilience level of the transgenic plants when compared with wild-type plants. KEY RESULTS: Overexpression of Scdr2 enhanced germination rates in tobacco seeds under drought and salinity conditions. Juvenile transgenic plants overexpressing Scdr2 and subjected to drought and salinity stresses showed higher photosynthesis levels, internal CO2 concentration and stomatal conductance, reduced accumulation of hydrogen peroxide in the leaves, no penalty for photosystem II and faster recovery after submission to both stress conditions. Respiration was not strongly affected by both stresses in the Scdr2 transgenic plants, whereas wild-type plants exhibited increased respiration rates. CONCLUSIONS: Scdr2 is involved in the response mechanism to abiotic stresses. Higher levels of Scdr2 enhanced resilience to salinity and drought, and this protection correlated with reduced oxidative damage. Scdr2 confers, at the physiological level, advantages to climate limitations. Therefore, Scdr2 is a potential target for improving sugarcane resilience to abiotic stress.

Brassinosteroids, the Sixth Class of Phytohormones: A Molecular View from the Discovery to Hormonal Interactions in Plant Development and Stress Adaptation.

Phytohormones are natural chemical messengers that play critical roles in the regulation of plant growth and development as well as responses to biotic and abiotic stress factors, maintaining plant homeostasis, and allowing adaptation to environmental changes. The discovery of a new class of phytohormones, the brassinosteroids (BRs), almost 40 years ago opened a new era for the studies of plant growth and development and introduced new perspectives in the regulation of agronomic traits through their use in agriculture. BRs are a group of hormones with significant growth regulatory activity that act independently and in conjunction with other phytohormones to control different BR-regulated activities. Genetic and molecular research has increased our understanding of how BRs and their cross-talk with other phytohormones control several physiological and developmental processes. The present article provides an overview of BRs' discovery as well as recent findings on their interactions with other phytohormones at the transcriptional and post-transcriptional levels, in addition to clarifying how their network works to modulate plant growth, development, and responses to biotic and abiotic stresses.

Transgenic Nicotiana tabacum seeds expressing the Mycobacterium tuberculosis Alanine- and Proline-rich antigen.

The glycoprotein APA (Alanine- and Proline-rich Antigen, a 45/47 kDa antigen complex, Rv1860) is considered as a major immunodominant antigen secreted by M. tuberculosis. This antigen has proved to be highly immunogenic in experimental models and humans, presenting a significant potential for further development of a new vaccine for tuberculosis. Glycosylation plays a key role in the immunogenicity of the APA protein. Because plants are known to promote post-translational modification such as glycosylation and to be one of the most economic and safe hosts for recombinant protein expression, we have over expressed the APA protein in transgenic tobacco plants aiming to produce a glycosylated version of the protein. Seeds are known to be a well-suited organ to accumulate recombinant proteins, due to low protease activity and higher protein stability. We used a seed-specific promoter from sorghum, a signal peptide to target the protein to the endoplasmic reticulum and ultimately in the protein storage vacuoles. We show that the recombinant protein accumulated in the seeds had similar isoelectric point and molecular weight compared with the native protein. These findings demonstrate the ability of tobacco plants to produce glycosylated APA protein, opening the way for the development of secure, effective and versatile vaccines or therapeutic proteins against tuberculosis.

pGVG: a new Gateway-compatible vector for transformation of sugarcane and other monocot crops.

The successful development of genetically engineered monocots using Agrobacterium-mediated transformation has created an increasing demand for compatible vectors. We have developed a new expression vector, pGVG, for efficient transformation and expression of different constructs for gene overexpression and silencing in sugarcane. The pCAMBIA2300 binary vector was modified by adding Gateway recombination sites for fast gene transfer between vectors and the maize polyubiquitin promoter Ubi-1 (ZmUbi1), which is known to drive high gene expression levels in monocots. Transformation efficiency using the pGVG vector reached up to 14 transgenic events per gram of transformed callus. Transgenic plants expressing the ß-glucuronidase (GUS) reporter gene from pGVG showed high levels of GUS activity. qRT-PCR evaluations demonstrated success for both overexpression and hairpin-based silencing cassettes. Therefore, pGVG is suitable for plant transformation and subsequent applications for high-throughput production of stable transgenic sugarcane. The use of an expression cassette based on the ZmUbi1 promoter opens the possibility of using pGVG in other monocot species.