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The role of WNT/ß-catenin signalling in mouse neocortex development remains ambiguous. Most studies demonstrate that WNT/ß-catenin regulates progenitor self-renewal but others suggest it can also promote differentiation. Here we explore the role of WNT/STOP signalling, which stabilizes proteins during G2/M by inhibiting glycogen synthase kinase (GSK3)-mediated protein degradation. We show that mice mutant for cyclin Y and cyclin Y-like 1 (Ccny/l1), key regulators of WNT/STOP signalling, display reduced neurogenesis in the developing neocortex. Specifically, basal progenitors, which exhibit delayed cell cycle progression, were drastically decreased. Ccny/l1-deficient apical progenitors show reduced asymmetric division due to an increase in apical-basal astral microtubules. We identify the neurogenic transcription factors Sox4 and Sox11 as direct GSK3 targets that are stabilized by WNT/STOP signalling in basal progenitors during mitosis and that promote neuron generation. Our work reveals that WNT/STOP signalling drives cortical neurogenesis and identifies mitosis as a critical phase for neural progenitor fate.
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Mitosis , Neocórtex/embriología , Neocórtex/metabolismo , Neurogénesis , Vía de Señalización Wnt , Secuencia de Aminoácidos , Animales , Biomarcadores , Ciclo Celular , Diferenciación Celular/genética , Ciclinas/genética , Ciclinas/metabolismo , Embrión de Mamíferos , Técnica del Anticuerpo Fluorescente , Expresión Génica , Glucógeno Sintasa Quinasa 3/metabolismo , Inmunohistoquímica , Ratones , Ratones Noqueados , Mitosis/genética , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , Neurogénesis/genética , Fosforilación , Factores de Transcripción SOXC/genética , Factores de Transcripción SOXC/metabolismoRESUMEN
A survey of Diaporthe/Phomopsis Complex (DPC) species was carried out on 479 asymptomatic soybean (Glycine max (L.) Merrill) seed samples collected from commercial soybean fields in the states of Santa Catarina (20 counties) and Rio Grande do Sul (41 counties), in the 2020/21 (n=186), 2021/22 (n=138) and 2022/23 (n=155) seasons from 120 cultivars. The seeds were provided by seed producers who collected according to the sampling standard of the Ministry of Agriculture, Livestock and Food Supply. From each sample received, 200 symptomless seeds were randomly sorted out. The seeds were surface disinfected by immersion in a sodium hypochlorite solution (1%) for two minutes and placed on Potato Dextrose Agar (PDA). The plates were incubated for 7 days at 23°C with a photoperiod of 12-h. The average prevalence of 73.7% of DPC-infected seeds. Colonies were isolated by transferring mycelial tips to PDA and incubating for 14 days at 25ºC in a 12-h photoperiod. One colony (isolate MEMR0500) had morphological characteristics similar to those reported in Lopez-Cardona (2021). This isolate had a floccose, dense colony ranging from grayish beige to brown with greenish regions and black globose pycnidia (3 to 4 pycnidia/cm²). Alpha-conidia, 5.1 to 7.0 µm x 1.5 to 2.8 µm, were observed after 30 days and were hyaline, aseptate and fusiform (Figure S1). No beta-conidia were observed. Soybean plants of cultivars BMX Cromo IPRO, BMX Zeus IPRO, BRS 5804 RR, FPS 1867 IPRO and NEO 750 IPRO were tested for pathogenicity using the toothpick inoculation method (Siviero and Menten 1995). Non-colonized toothpicks served as a negative control. Plants were incubated for four days at 25°C and 90% relative humidity. Elongated 1.0 to 2.5 cm x 0.5 to 0.9 cm lesions gray-brown/reddish-brown with a depressed center were observed in all inoculated cultivars. The fungus was reisolated and the characteristics of the colonies were identical to those previously isolated. For molecular characterization, DNA was extracted from the mycelia using the CTAB method (Doyle and Doyle 1990). End-point PCR was performed using GoTaq® Flexi DNA Polymerase (Promega, USA) and primer pairs, ITS-4F/ITS-5, T2/Bt2b and EF1-728F/EF1-986R to amplify the internal transcribed spacer (ITS) (Costamilan et al. 2008), ß-tubulin (TUB2) (Glass and Donaldson 1995), and translation elongation factor 1-α (TEF1) (Carbone and Kohn 1999) genes, respectively. The amplified fragments were sequenced and submitted to blast search (https://blast.ncbi.nlm.nih.gov/Blast.cgi) with the sequences available in GenBank. The fragment from ITS (accession number OR912979) showed 99.8% (549/582 bp) identity with Diaporthe ueckeri Udayanga & Castl. [as 'ueckerae'] [syn. D. miriciae R.G. Shivas, S.M. Thomps. & Y.P. Tan] isolate FAU656 (Ac. N. KJ590726). The sequence of TEF (Ac. N. PP372869) showed 99.7% (339/355 bp) identity with D. ueckeri FAU656 (Ac. N. KJ590747), and of TUB (Ac. N. PP372870) showed 98.9% (436/536 bp) identity with D. ueckeri FAU656 (Ac. N. KJ610881). A phylogenetic tree with amplified sequences of each gene and the corresponding representative sequences from the DPC was constructed in MEGA X (Kumar et al. 2018). The MEMR0500 isolate was clustered only with the D. ueckeri clade, confirming the identity of the fungus (Figure S2). In Brazil, this is the first report of the association of this pathogen with soybean seeds. In other countries, this pathogen has been identified as the causal agent of stem canker (Mena et al. 2020; Lopez-Cardona et al. 2021). Further research is needed to analyze the risk of this seed-associated pathogen.
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Adrenal glands are zonated endocrine organs that are essential in controlling body homeostasis. How zonation is induced and maintained and how renewal of the adrenal cortex is ensured remain a mystery. Here we show that capsular RSPO3 signals to the underlying steroidogenic compartment to induce ß-catenin signaling and imprint glomerulosa cell fate. Deletion of RSPO3 leads to loss of SHH signaling and impaired organ growth. Importantly, Rspo3 function remains essential in adult life to ensure replenishment of lost cells and maintain the properties of the zona glomerulosa. Thus, the adrenal capsule acts as a central signaling center that ensures replacement of damaged cells and is required to maintain zonation throughout life.
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Corteza Suprarrenal/fisiología , Diferenciación Celular/genética , Transducción de Señal/genética , Trombospondinas/metabolismo , Corteza Suprarrenal/citología , Animales , Proliferación Celular , Embrión de Mamíferos , Eliminación de Gen , Regulación del Desarrollo de la Expresión Génica/genética , Homeostasis/genética , Masculino , Ratones , Trombospondinas/genética , Zona Glomerular/citología , Zona Glomerular/metabolismo , beta Catenina/metabolismoRESUMEN
This study analyzed and compared the physicochemical and mechanical properties of preheated resin composite with light-cured resin cement for luting indirect restorations. 210 specimens of resin cement/resin composite were prepared according to preheating treatment heated (Htd) or not (NHtd). Light-cured resin cement (Variolink Veneer, Ivoclar), and resin composite (Microhybrid-Z100, 3 M; Nanohybrid-Empress direct, Ivoclar; and Bulk fill-Filtek One, 3 M) were used (n = 10). Resin cement specimens were not preheated. The response variables were (n = 10): film thickness, microhardness, liquid sorption and solubility. Data were analyzed by 2-way ANOVA and Tukey HSD post-test (α = 0.05). Bulk fill NHtd resin had the highest film thickness values (p < 0.001). Microhybrid and nanohybrid Htd resins had the smallest thicknesses and did not differ from the cement (p > 0.05). The highest microhardness values were found for Bulk fill NHtd and Bulk fill Htd resins. The nanohybrid and microhybrid Htd resins showed the lowest microhardness values, with no difference in cement (p > 0.05). For liquid sorption, there was no significant difference between the groups (p = 0.1941). The microhybrid Htd resin showed higher solubility values than the other materials (p = 0.0023), but it did not differ statistically from resin cement (p > 0.05). Preheating composite resins reduced the film thickness. After heating, nanohybrid and Bulk fill resins retained stable microhardness, sorption, and solubility values.
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Cerámica , Resinas Compuestas , Ensayo de Materiales , Solubilidad , Resinas Compuestas/química , Cerámica/química , Cementos de Resina/química , Propiedades de Superficie , Dureza , Calor , Fenómenos QuímicosRESUMEN
Coordinated development of muscles, tendons, and their attachment sites ensures emergence of functional musculoskeletal units that are adapted to diverse anatomical demands among different species. How these different tissues are patterned and functionally assembled during embryogenesis is poorly understood. Here, we investigated the morphogenesis of extraocular muscles (EOMs), an evolutionary conserved cranial muscle group that is crucial for the coordinated movement of the eyeballs and for visual acuity. By means of lineage analysis, we redefined the cellular origins of periocular connective tissues interacting with the EOMs, which do not arise exclusively from neural crest mesenchyme as previously thought. Using 3D imaging approaches, we established an integrative blueprint for the EOM functional unit. By doing so, we identified a developmental time window in which individual EOMs emerge from a unique muscle anlage and establish insertions in the sclera, which sets these muscles apart from classical muscle-to-bone type of insertions. Further, we demonstrate that the eyeballs are a source of diffusible all-trans retinoic acid (ATRA) that allow their targeting by the EOMs in a temporal and dose-dependent manner. Using genetically modified mice and inhibitor treatments, we find that endogenous local variations in the concentration of retinoids contribute to the establishment of tendon condensations and attachment sites that precede the initiation of muscle patterning. Collectively, our results highlight how global and site-specific programs are deployed for the assembly of muscle functional units with precise definition of muscle shapes and topographical wiring of their tendon attachments.
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Músculos Oculomotores/embriología , Músculos Oculomotores/crecimiento & desarrollo , Tretinoina/metabolismo , Animales , Tejido Conectivo/fisiología , Desarrollo Embrionario , Ojo , Imagenología Tridimensional/métodos , Ratones/embriología , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Morfogénesis , Transducción de Señal , Tendones/fisiología , Tretinoina/fisiologíaRESUMEN
Plant-derived polyphenols are naturally occurring compounds widely distributed in plants. They have received greater attention in the food and pharmaceutical industries due to their potential health benefits, reducing the risk of some chronic diseases due to their antioxidant, anti-inflammatory, anticancer, cardioprotective, and neuro-action properties. Polyphenolic compounds orally administered can be used as adjuvants in several treatments but with restricted uses due to chemical instability. The review discusses the different structural compositions of polyphenols and their influence on chemical stability. Despite the potential and wide applications, there is a need to improve the delivery of polyphenolics to target the human intestine without massive chemical modifications. Oral administration of polyphenols is unfeasible due to instability, low bioaccessibility, and limited bioavailability. Nano-delivery systems based on polysaccharides (starch, pectin, chitosan, and cellulose) have been identified as a viable option for oral ingestion, potentiate biological effects, and direct-controlled delivery in specific tissues. The time and dose can be individualized for specific diseases, such as intestinal cancer. This review will address the mechanisms by which polysaccharides-based nanostructured systems can protect against degradation and enhance intestinal permeation, oral bioavailability, and the potential application of polysaccharides as nanocarriers for the controlled and targeted delivery of polyphenolic compounds.
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Human behaviour is a dense longitudinal multi-featured measure that directly impacts the health of individuals in the short and long terms. Therefore, issues usually emerge from the insistence on performing risky behaviours, such as smoking or eating fast foods, which continuously increase the gap between current and beneficial health states. This paper introduces the term "health debt" as an economic metaphor to represent the quantification of this gap in domains such as sleep, contributing to physical and mental health states. Then, we present a theoretical framework that relies on behaviour change recommendations to quantify this debt. The practical instantiation of this framework relies on passively assessed sleep related data via personal wearable devices, and uses of an attention-based predictive model as the fitness function of a genetic algorithm that acts as a recommender. We evaluate this proposal by means of a case example aimed at improving the sleep duration of individuals. Results show, for example, that the use of individual rather than generic datasets produces more accurate models. At the same time, the use of constraints on the variability of behaviours features generates more feasible recommendations. These foundations open new research opportunities to support the adoption of preventive medicine based on longitudinal wearable passive data analysis.
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Aprendizaje Profundo , Dispositivos Electrónicos Vestibles , Humanos , Sueño , Salud Mental , Ejercicio FísicoRESUMEN
BACKGROUND: Magnetic resonance diffusion tensor imaging (MR-DTI) has been increasingly applied for carpal tunnel syndrome (CTS) diagnosis, but relatively little is known about the effect of CTS treatment on median nerve (MN) integrity and functional outcome prediction. PURPOSE: To assess how structural changes in MR-DTI of the MN correlates with symptom severity, functional status, and electrophysiological parameters in patients suffering from CTS before and after decompression surgery. MATERIAL AND METHODS: Nine wrists were prospectively enrolled to perform MR-DTI pre- and postoperatively. The apparent diffusion coefficients (ADC) and fractional anisotropy (FA) of the MN were examined in three different regions-distal radioulnar joint, pisiform bone, and hamate bone-and correlated with clinical and electrophysiological parameters. RESULTS: Postoperatively, mean Boston Carpal Tunnel Questionnaire scores decreased 1.55 points (range = 0.08-3; P = 0.0172) and 1.01 points (-0.13 to 1.88; P = 0.0381) in the symptomatic and functional domains, respectively. Postoperative clinical improvement was reflected in proximal FA elevation (P = 0.0078), but not in diffusivity in comparison to baseline examination. Preoperative electrophysiological parameters were correlated with a reduction in the pre- (sensory latencies [rho = -0.6826; P = 0.0312]) and postoperative (motor latencies [rho = -0.7488; P = 0.0325]) distal FA values. Higher sensory amplitudes indicated higher postoperative proximal FA values (rho = 0.7618; P = 0.0280) ââand lower postoperative proximal ADC values (rho = -0.9047; P = 0.0020). CONCLUSION: Our study demonstrated that pre- and postoperative proximal FA values are useful biomarkers for the structural evaluation of the MN in patients with CTS. Symptomatic improvement can be better predicted by analyzing FA changes.
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Síndrome del Túnel Carpiano , Humanos , Síndrome del Túnel Carpiano/diagnóstico por imagen , Síndrome del Túnel Carpiano/cirugía , Imagen de Difusión Tensora/métodos , Pronóstico , Biomarcadores , DescompresiónRESUMEN
Brazil is the largest producer of soybean [Glycine max (L.) Merrill], cultivated in diverse environments and systems. This scenario can contribute to emergence of new diseases or increase the severity of secondary diseases. In March 2023, elliptical to circular, brownish lesions, 5.2-6.1 cm length and 1.1-1.5 cm width, with salmon-colored masses of conidia in the center of the lesions, were observed on the stems of soybean cultivar 'CZ 16B17 IPRO', in the municipality of Campos Novos, Santa Catarina, Brazil (27º25'19''S and 51º14'14''05W). The presence of 210-355 µm length and 210-232 µm width acervuli was rare, with arrows larger than the mass of conidia (Figure S1). Fragments of the infected tissues were cut, disinfected and placed in Petri dishes containing Potato Dextrose Agar (PDA) or V8-agar medium and maintained at 23 ± 2ºC and a photoperiod of 12 h dark-light cycle. After 13 days, the development of grayish-white colonies was observed on both culture media, with the formation of a mass of septate hyaline, oblong, cylindrical conidia, 13.3-15.3 µm length and 2.9-3.5 µm width, with obtuse ends. One pure monosporic isolate was selected, isolate CF1. The presence of sexual structures was observed on PDA after 13 days and in V8 after 15-20 days. Perithecia were dark brown and globose, either immersed in the culture medium or on the surface between the mycelia. Inside of perithecia, unitunicate, clavate, and cymbiform asci, 39.1-61.0 µm length and 9.6-11.7 µm width were observed, containing eight spindle-shaped and slightly curved ascospores with rounded tips 13.8-18.3 µm length and 3.0-4.2 µm width (Figure S1). Pathogenicity tests were performed on young soybean plants at V1 phenological growth stage in four repetitions. PDA disks, 7mm in diameter, with growth mycelium were placed on stems while using uninfected PDA disks as a control. Plants were incubated in a chamber at 25 ± 2°C and 90% relative humidity. Anthracnose lesions were observed only on the stems of the inoculated plants. The same pattern of symptoms was observed on the stems, and the fungus were reisolated on PDA. The colony and morphological characteristics were identical to the previously isolated fungus. For molecular characterization, the growth mycelia were collected, macerated in liquid nitrogen, and DNA was extracted using the method Doyle and Doyle (1990) with CTAB. End-point PCR was performed using the GoTaq® Flexi DNA Polymerase (Promega, USA) and the primers, ITS-1F/ITS-4, T1/Bt2b, CL1C/CL2C, GDF/GDR, and SODglo2-F/SODglo2-R (Weir et al. 2012) for the amplification of internal transcribed spacer (ITS), ß-tubulin (TUB2), calmodulin (CAL), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and superoxide dismutase (SOD), respectively. Amplified fragments were sequenced and compared with the available sequences in the Genbank (www.ncbi.nlm.nih.gov/genbank/). The sequences of all five-genes (Accession numbers OR883777, OR891749, OR891750, OR891751 and OR891752, respectively) of the isolate CF1 characterized in this study showed 99% nucleotide identities whith the stand isolate ICMP 18581 of Colletotrichum fructicola. A phylogenetic tree was constructed in MEGA X (Kumar et al. 2021), containing the amplified and concatenated sequences and representative species from the Colletotrichum gloeosporioides complex. The isolate grouped only with C. fructicola clade, confirming the identity of the fungus (Figure S2). To our knowledge, this is the first study reporting the infection of C. fructicola in soybeans in Brazil, which has already been reported in China (Xu et al. 2023).
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Adaptive AI for context and activity recognition remains a relatively unexplored field due to difficulty in collecting sufficient information to develop supervised models. Additionally, building a dataset for human context activities "in the wild" demands time and human resources, which explains the lack of public datasets available. Some of the available datasets for activity recognition were collected using wearable sensors, since they are less invasive than images and precisely capture a user's movements in time series. However, frequency series contain more information about sensors' signals. In this paper, we investigate the use of feature engineering to improve the performance of a Deep Learning model. Thus, we propose using Fast Fourier Transform algorithms to extract features from frequency series instead of time series. We evaluated our approach on the ExtraSensory and WISDM datasets. The results show that using Fast Fourier Transform algorithms to extract features performed better than using statistics measures to extract features from temporal series. Additionally, we examined the impact of individual sensors on identifying specific labels and proved that incorporating more sensors enhances the model's effectiveness. On the ExtraSensory dataset, the use of frequency features outperformed that of time-domain features by 8.9 p.p., 0.2 p.p., 39.5 p.p., and 0.4 p.p. in Standing, Sitting, Lying Down, and Walking activities, respectively, and on the WISDM dataset, the model performance improved by 1.7 p.p., just by using feature engineering.
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Algoritmos , Caminata , Humanos , Actividades Humanas , Movimiento , Factores de TiempoRESUMEN
The recognition of human activities (HAR) using wearable device data, such as smartwatches, has gained significant attention in the field of computer science due to its potential to provide insights into individuals' daily activities. This article aims to conduct a comparative study of deep learning techniques for recognizing activities of daily living (ADL). A mapping of HAR techniques was performed, and three techniques were selected for evaluation, along with a dataset. Experiments were conducted using the selected techniques to assess their performance in ADL recognition, employing standardized evaluation metrics, such as accuracy, precision, recall, and F1-score. Among the evaluated techniques, the DeepConvLSTM architecture, consisting of recurrent convolutional layers and a single LSTM layer, achieved the most promising results. These findings suggest that software applications utilizing this architecture can assist smartwatch users in understanding their movement routines more quickly and accurately.
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Actividades Cotidianas , Aprendizaje Profundo , Humanos , Reconocimiento en Psicología , Benchmarking , MovimientoRESUMEN
Several studies have proved that glial cells, as well as neurons, play a role in pain pathophysiology. Most of these studies have focused on the contribution of central glial cells (e.g., microglia and astrocytes) to neuropathic pain. Likewise, some works have suggested that peripheral glial cells, particularly satellite glial cells (SGCs), and the crosstalk between these cells and the sensory neurons located in the peripheral ganglia, play a role in the phenomenon that leads to pain. Nonetheless, the study of SGCs may be challenging, as the validity of studying those cells in vitro is still controversial. In this study, a research protocol was developed to examine the potential use of primary mixed neuronal-glia cell cultures obtained from the trigeminal ganglion cells (TGCs) of neonate mice (P10-P12). Primary cultures were established and analyzed at 4 h, 24 h, and 48 h. To this purpose, phase contrast microscopy, immunocytochemistry with antibodies against anti-ßIII-tubulin and Sk3, scanning electron microscopy, and time-lapse photography were used. The results indicated the presence of morphological changes in the cultured SGCs obtained from the TGCs. The SGCs exhibited a close relationship with neurons. They presented a round shape in the first 4 h, and a more fusiform shape at 24 h and 48 h of culture. On the other hand, neurons changed from a round shape to a more ramified shape from 4 h to 48 h. Intriguingly, the expression of SK3, a marker of the SGCs, was high in all samples at 4 h, with some cells double-staining for SK3 and ßIII-tubulin. The expression of SK3 decreased at 24 h and increased again at 48 h in vitro. These results confirm the high plasticity that the SGCs may acquire in vitro. In this scenario, the authors hypothesize that, at 4 h, a group of the analyzed cells remained undifferentiated and, therefore, were double-stained for SK3 and ßIII-tubulin. After 24 h, these cells started to differentiate into SCGs, which was clearer at 48 h in the culture. Mixed neuronal-glial TGC cultures might be implemented as a platform to study the plasticity and crosstalk between primary sensory neurons and SGCs, as well as its implications in the development of chronic orofacial pain.
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Since 1948, pale yellow wheat spike have been reported in southern Brazil. This symptom was associated with tenuiviruses due to the observation of cytoplasmic inclusions constituted by a mass of filamentous particles (7-10 nm in diameter) with indeterminate length, identical to those found in "leaf dip" preparations. Such symptoms are still seen in wheat crops; however, there is a lack of information regarding this pathosystem. Decades after the first report, the first sequences of wheat white spike virus were characterized. Wheat plants with symptoms such as pale yellowing, chlorotic streaks, and leaf mosaic were collected in Paraná State, Southern Brazil. High-throughput sequencing was used to determine the nearly complete nucleotide sequence of the viral genome. The genome is composed of five RNAs with a total size of 18,129 nucleotides, with eight open reading frames (ORFs). The virus identified in this study can be included in a new species in the family Phenuiviridae, genus Tenuivirus, and we have tentatively named this virus "wheat white spike virus".
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Enfermedades de las Plantas/virología , Tenuivirus , Triticum/virología , Brasil , Filogenia , Tenuivirus/clasificaciónRESUMEN
Five elephant garlic plants (Allium ampeloprasum L.) showing leaf symptoms of chlorotic streaks and mosaic (Figure 1A and B) were collected, in September 2021, in an experimental area in municipality of Rio do Sul (27°11'07"S, 49°39'39"W), State of Santa Catarina, Brazil. Total RNA was extracted using TRIzol® reagent (Invitrogen, USA), according to the manufacturer's instructions to investigate viral infection. The RNA from all five plants were pooled into a single sample for cDNA library construction with the TruSeq Stranded Total RNA with Ribo-Zero Plant (Illumina) kit, which was then sequenced on the Illumina HiSeq2500 platform (Proteimax Biotechnology LTDA). After high throughput sequencing (HTS), 49 million raw reads (each 151nt) were generated. They were trimmed with the BBduk tool and de novo assembled with the Tadpole assembler tool (Geneious Software version 2022). A total of 28,345 contigs were generated and searched against the NCBI virus genome database using BLASTn and BLASTx, with positive results for two potyviruses, leek yellow stripe virus (LYSV), onion yellow dwarf virus (OYDV), and the putative polerovirus allium polerovirus A (APVA). The trimmed reads were mapped with the BBmap tool (Bushell 2014), using reference sequences for LYSV (NC_004011), OYDV (NC_005029), and APVA isolate Won (MH898527). A total of 806,060 reads were mapped, resulting in the nearly complete genome of LYSV (isolate RDS22-2, 10,268 bp, ON565071), which shared the highest (89.41%) nucleotide (nt) identity with LYSV isolate MG (KP258216). The nearly complete genome of OYDV (isolate RDS22-1, 10,519 bp, ON565070) was assembled using 311,467 reads, being 90.21% nt identical to OYDV isolate G-118 (KF632714). The APVA genome (isolate RDS22-3, 4,367 bp, ON565072, Figure 1C) was assembled from 116,303 reads and it shared the highest (90.73%) nt identity with APVA isolate Won. Subsequently, each sample was RT-PCR screened separately for potyviruses and poleroviruses, using the generic primer pairs NIb2F/NIb3R (Zheng et al., 2010) and Pol-G-F/Pol-G-R (Knierim et al., 2010), respectively. Amplified DNA fragments with approximately 350 bp and 1000 bp were obtained for potyviruses and poleroviruses, respectively, and were sent for Sanger sequencing (ACTGene, Alvorada, Brazil). The Sanger derived partial sequences shared 98 to 100% nt identities with corresponding HTS-derived sequences. The most common virus was LYSV, which was found in three of the five tested samples, whereas OYDV and APVA were only found in one sample each. The plants were also screened with specific primers for each virus, and none of the samples revealed mixed infections. Elephant garlic is primarily utilized for industrial garlic production in several countries, and it is now being researched in Brazil for the same purpose. It can be observed from this study that elephant garlic is susceptible to two of the most common viruses in garlic (LYSV and OYDV), which must be considered in the future while developing resistant varieties or in using thermotherapy and shoot tip/meristem culture to recover virus-free cultivars. LYSV and OYDV have already been described in Brazil infecting Allium sativum (Kitajima 2020). The only complete APVA sequence available is from China (Isolate Won), but no further characterization of the virus has been performed and published. The occurrence of this virus in Brazil highlights the importance of further research to obtain a more robust virus characterization.
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Maize (Zea mays L.) is the main cereal food of humans and animals in Brazil. In 2020 and 2021, a severe infestation of corn leafhoppers (Dalbulus maidis; Hemiptera: Cicadellidae) was observed in Santa Catarina State (South of Brazil). Subsequently, symptoms of chlorotic stripes limited in leaf veins started to appear in maize plants. Given the similarity of symptoms and the presence of high populations of corn leafhoppers in corn production areas, 30 plants in reproductive stage showing systemic symptoms were collected in summer and autumn from commercial fields of five municipalities in Santa Catarina: Campos Novos (27°23'18.0"S, 51°12'52.7"W), Lages (27°47'17.8"S, 50°18'16.9"W), Mafra (26°06'42"S, 49°48'25"W), Fraiburgo (27°01'36"S, 50°55'19"W), and Abelardo Luz (26°34'02"S, 52°20'02"W). The young leaves of these samples were used for molecular analyses targeting the maize rayado fino virus (MRFV; Tymoviridae: Marafivirus). Total nucleic acids were extracted using TRIzol® (Invitrogen, USA), following the manufacturer's instructions. These were used as a template for cDNA synthesis with the enzyme MMLV-RT (Promega, USA), following the manufacturer's instructions. The polymerase chain reaction (PCR) was performed using Gotaq® DNA polymerase (Promega, USA) and MRFV-09/MRFV-10 primers (Hammond et al. 1997). All PCR products were subjected to electrophoresis in 1% agarose gel and were visualized under ultraviolet light. Twenty-eight of the 30 tested plants were MRFV-positive, showing a fragment with an expected size of ~633 bp. To confirm our results, all MRFV-positive samples were sent for sequencing (GenBank accession numbers OM763708 - OM763710 and ON730784 - ON730806) and submitted to BLASTn search (https://blast.ncbi.nlm.nih.gov/Blast.cgi), resulting in identities ranging from 96.21% to 99.21% with the isolate "Brazil 26" of MRFV, which was detected in 2005 in São Paulo, Brazil (GenBank accession nº: AF186178) (Hammond and Bedendo 2005). A second set of primers was used to validate the first PCR, confirming MRFV infection (data not shown).Moreover, whitish streaks and leaf reddening were observed on the leaves of some plants; therefore, the identification for phytoplasmas (Candidatus Phytoplasma asteris) and spiroplasmas (Spiroplasma kunkelii) from the corn stunt complex was performed. For this, previously extracted nucleic acids from each sample were used as templates for a multiplex PCR using the primers CSSR6/CSSF2 and R16F2n/R16R2 (Gundersen and Lee 1996; Barros et al. 2001). Two plants were infected with only spiroplasma, 17 samples were infected with Spiroplasma and MRFV, and three samples were infected by these three pathogens. An increasing incidence of corn stunt has been observed in commercial fields in Santa Catarina in recent years. Mollicutes are commonly found and mostly studied as causal agents of corn stunt disease. On the contrary, despite being present in Brazil since the 1970s, the virus is less studied because its contribution to the corn stunt complex is still unknown (Hammond and Bedendo 2001). In this report, indications that the virus is expanding to different regions in southern Brazil were observed, which raises an opportunity for further evaluation and its consideration in monitoring programs. Moreover, to the best of our knowledge, this is the first report of MRFV in Santa Catarina, Brazil.
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Technological development is key for national strategies to cope with the Paris Agreement's goals. Technology Needs Assessments (TNAs) aim to identify, prioritize, and diffuse climate change mitigation and/or adaptation technologies in developing countries. Their methodology includes a multi-criteria decision analysis (MCDA) framework but, although many countries already conducted a TNA, literature lacks discussions on country-specific processes for a TNA, as it usually follows a one-size-fits-all approach. This paper provides empirical evidence on the importance of country-driven processes that help shaping international programmes into country-specific needs and capabilities. It presents lessons learned from a tailored process for identification, prioritization, and selection of mitigation technologies in the scope of a TNA project for Brazil, an exceptional case of a developing country with strong capacity in integrated assessment modelling (IAM) scenarios for guiding its climate strategies. A previous IAM scenario result allowed pre-selecting technologies in six key economic sectors, while other TNAs prioritized no more than three. This allowed the elaboration of an overall ranking from the MCDA, in contrast to sectoral rankings that are mostly employed in other countries' TNAs. The overall ranking serves not only as a basis for the selection of priority technologies but also provides information on the integrated innovations framework for climate technologies in the country. Further specific findings of the tailored Brazilian TNA approach are discussed in the paper in order to call for the importance that a technology transfer project should not only be country-driven but also conducted through a country-specific process. Supplementary Information: The online version contains supplementary material available at 10.1007/s11027-022-10025-6.
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Rice (Oryza sativa L.) is an important food crop for humanity, being cultivated in tropical and temperate regions of the world. This study reports the nearly complete genome sequences of four Brazilian rice stripe necrosis virus (RSNV) isolates. The nucleotide sequences of the RNA1 and RNA2 genome segments of these Brazilian isolates were 96.5 to 99.9% identical, indicating their close phylogenetic relationship to each other. Phylogeny and recombination analysis indicated that the genome of one of the isolates consisted of RNA segments of different origins, suggesting that a reassortment event had occurred.
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Oryza/virología , Virus de Plantas/genética , Brasil , FilogeniaRESUMEN
Schistosomiasis is a parasitic disease that affects more than 250 million people. The treatment is limited to praziquantel and the control of the intermediate host with the highly toxic molluscicidal niclosamide. Marine algae are a poorly explored and promising alternative that can provide lead compounds, and the use of multivariate analysis could contribute to quicker discovery. As part of our search for new natural compounds with which to control schistosomiasis, we screened 45 crude extracts obtained from 37 Brazilian seaweed species for their molluscicidal activity against Biomphalaria glabrata embryos and schistosomicidal activities against Schistosoma mansoni. Two sets of extracts were taxonomically grouped for metabolomic analysis. The extracts were analyzed by GC-MS, and the data were subjected to Pattern Hunter and Pearson correlation tests. Overall, 22 species (60%) showed activity in at least one of the two models. Multivariate analysis pointed towards 3 hits against B. glabrata veliger embryos in the Laurencia/Laurenciella set, 5 hits against B. glabrata blastula embryos, and 31 against S. mansoni in the Ochrophyta set. Preliminary annotations suggested some compounds such as triquinane alcohols, prenylated guaianes, dichotomanes, and xenianes. Despite the putative identification, this work presents potential candidates and can guide future isolation and identification.
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Biomphalaria/efectos de los fármacos , Bioprospección , Descubrimiento de Drogas , Moluscocidas/farmacología , Schistosoma mansoni/efectos de los fármacos , Esquistosomiasis mansoni/prevención & control , Esquistosomicidas/farmacología , Algas Marinas/metabolismo , Animales , Biomphalaria/parasitología , Brasil , Metaboloma , Metabolómica , Moluscocidas/aislamiento & purificación , Schistosoma mansoni/crecimiento & desarrollo , Esquistosomiasis mansoni/parasitología , Esquistosomicidas/aislamiento & purificaciónRESUMEN
The hop (Humulus lupulus L.) is a dioecious perennial climbing plant grown commercially worldwide. Wild hops are widely distributed throughout the Northern Hemisphere, Europe, Asia, and North America (Neve, 1991). In the Southern Hemisphere, some of the leading hop-producing countries include South Africa, Australia, and New Zealand. Brazil began hop production less than 5 years ago. In January 2019, amphigenous white powdery circular fungal colonies were observed on the leaves and stems of hop plants (cultivar Chinook) within a 900m2 hop garden in Lages municipality, Santa Catarina State, southern Brazil. The incidence of the disease was present on almost 100 per cent of "Chinook" cultivar plants and diseased foliage was collected to identify the pathogen and used to inoculate healthy plants. Hop powdery mildew lesions with hyaline and septate mycelium with chains of unicellular conidia (n =100) hyaline, barrel-shaped, mean of length/width ± standard deviation 25-27 × 13-18 µm ± 0.980, with fibrosin bodies, and conidiophores erect with cylindrical foot cells, were visible within 10 days. The causal agent was identified as Podosphaera macularis (Wallr.:Fr.) Lind (synonym S. humuli (DC.) Burrill) on the basis of conidial shape, size and host range (Royle 1978; Braun 1987; Mahaffee et al., 2009), complemented with the present molecular analysis. Chasmothecia have not been observed in the field to date. A conidial suspension of 200 ml at concentration of 1.4 x 105 was mixed with 5ul of Tween® 20 for the pathogenicity assay. Ten plants of 9-month-old of hop "Chinook" cultivar, were inoculated with 5 ml of the conidial suspension using a manual spray. The control plot was only sprayed with water. The inoculated plants were maintained at 22ºC ± 1ºC with a 12-hour photoperiod and 65% relative humidity. White mycelia were visible first on the adaxial leaf surfaces of the inoculated younger leaves after 10 days and the disease severity reached between 2 to 5%. No symptoms were observed at the control plot. P. macularis infected most aerial plant tissues of the inoculated plants and caused approximately 50% of cones losses. P. macularis conidia were collected from the infected leaf tissue with a sterile soft camel-hair brush and DNA was extracted using a Wizard Genomic DNA extraction kit. The primers ITS1/ITS4 (White et al., 1990) were used to amplified and sequenced a fragment of the ITS region. PCR products were subjected to Sanger Sequencing to confirm sample species. The resulting 522-bp sequence was deposited into GenBank (accession n°. MN630490). BLASTn showed a 99.81% sequence identity with the CT1 isolate of P. macularis from H. lupulus (MH687414). The presence and identification of P. macularis in hop production regions is a new challenge to growers in Brazil. Research related to the knowledge of the disease cycle, epidemiology, and control strategies for the integrated management should be conducted, as there are no registered fungicides for powdery mildew on hop in Brazil. To our knowledge, this is the first report of P. macularis in Brazil, as well as in South America. References Braun, U. (1987) A Monograph of the Erysiphales (Powdery Mildews). J. Cramer, Berlin, German Democratic Republic. p 113. Mahaffee, W. F., Pethybridge, S.J., Gent, D.H (2009) Compendium of hop diseases and pests. The American Phytopathological Society Press, Saint Paul, Minnesota. Neve R. A (1991). Hops. Chapman and Hall: London. Royle, D. J (1978). Powdery mildew of the hop. Pages 381-409 in: The Powdery Mildews. D. M. Spencer, ed. Academic Press, New York. White, T. J., Bruns, T., Lee, S., and Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. pp. 315-322 in: PCR Protocols: A Guide to Methods and Applications. M. Innis, D. Gelfand, J. Sninsky, and T. White, eds. Academic Press, San Diego.
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PURPOSE: The cubital tunnel is limited anteriorly by the medial epicondyle (ME), laterally by the medial collateral ligament, and superiorly by Osborne's fascia and the cubital tunnel retinaculum. Previous studies were mostly dedicated to the roof of the cubital tunnel, in the way that the study of the groove for ulnar nerve and ME anatomy is relatively scarce in the literature. We sought to describe the radiological anatomy of the groove for ulnar nerve and ME in healthy volunteers with multiplanar computed tomography (CT). METHODS: We analyzed 3D CT images of 30 healthy volunteers (mean age 39 years, range 18-66 years). Nine variables were measured from the right elbow, including sizes, areas and angles in two different planes (coronal and axial). RESULTS: Mean ME width and length were 17.3 ± 3.5 mm and 31.7 ± 4.5 mm, respectively. According to categorical correlation studies, ME width (X) was deemed the most representative morphological characteristic because of the positive correlation to five other different anatomical measurements. A three-tiered anatomical classification was proposed based on data distribution. CONCLUSION: Large individual variation is found in the shape of ME, both in coronal and axial planes. The knowledge of individual osseous morphology is of great value potentially contributing to the surgical decision-making in patients affected by cubital tunnel syndrome.