RESUMO

Successful plant reproduction depends on the adequate development of flower organs controlled by cell proliferation and other processes. The SCI1 gene regulates cell proliferation and affects the final size of the female reproductive organ. To unravel the molecular mechanism exerted by SCI1 in cell proliferation control, we searched for its interaction partners through semi-in vivo pulldown experiments, uncovering a cyclin-dependent kinase, NtCDKG;2. Bimolecular fluorescence complementation (BiFC) and co-localization experiments showed that SCI1 interacts with NtCDKG;2 and its cognate NtCyclin L in nucleoli and splicing speckles. The screening of a yeast two-hybrid (Y2H) cDNA library using SCI1 as bait revealed a novel DEAD-box RNA helicase (NtRH35). The interaction between the NtCDKG;2-NtCyclin L complex, and NtRH35 was also shown. Subcellular localization experiments showed that SCI1, NtRH35, and the NtCDKG;2-NtCyclin L complex associate with each other within splicing speckles. The Y2H screening of NtCDKG;2 and NtRH35 identified the conserved spliceosome components U2a', NKAP, and CACTIN. This work presents SCI1 and its interactors NtCDKG;2-NtCyclin L complex, and NtRH35 as new spliceosome-associated proteins. Our findings reveal a network of interactions and suggest that SCI1 may regulate cell proliferation through the splicing process. This study provides new valuable insights into the intricate molecular pathways governing plant development.

RESUMO

Fragaria chiloensis is a Chilean native species that softens intensively during its ripening. Its softening is related to cell wall disassembly due to the participation of cell wall degrading enzymes. Softening of F. chiloensis fruit can be accelerated by ABA treatment which is accompanied by the increment in the expression of key cell wall degrading genes, however the molecular machinery involved in the transcriptional regulation has not been studied until now. Therefore, the participation of two MADS-box transcription factors belonging to different subfamilies, FchAGL9 and FchSHP, was addressed. Both TFs are members of type-II MADS-box family (MIKC-type) and localized in the nucleus. FchAGL9 and FchSHP are expressed only in flower and fruit tissues, rising as the fruit softens with the highest expression level at C3-C4 stages. EMSA assays demonstrated that FchAGL9 binds to CArG sequences of RIN and SQM, meanwhile FchSHP interacts only with RIN. Bimolecular fluorescence complementation and yeast two-hybrid assays confirmed FchAGL9-FchAGL9 and FchAGL9-FchSHP interactions. Hetero-dimer structure was built through homology modeling concluding that FchSHP monomer binds to DNA. Functional validation by Luciferase-dual assays indicated that FchAGL9 transactivates FchRGL and FchPG's promoters, meanwhile FchSHP transactivates those of FchEXP2, FchRGL and FchPG. Over-expression of FchAGL9 in C2 F. chiloensis fruit rises FchEXP2 and FchEXP5 transcripts, meanwhile the over-expression of FchSHP also increments FchXTH1 and FchPL; in both cases there is a down-regulation of FchRGL and FchPG. In summary, we provided evidence of FchAGL9 and FchSHP participating in the transcription regulation associated to F. chiloensis's softening.

Assuntos

Fragaria , Frutas , Regulação da Expressão Gênica de Plantas , Proteínas de Domínio MADS , Proteínas de Plantas , Frutas/genética , Frutas/metabolismo , Proteínas de Domínio MADS/genética , Proteínas de Domínio MADS/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fragaria/genética , Fragaria/metabolismo

RESUMO

A novel MADS-box transcription factor from Pinus radiata D. Don was characterized. PrMADS11 encodes a protein of 165 amino acids for a MADS-box transcription factor belonging to group II, related to the MIKC protein structure. PrMADS11 was differentially expressed in the stems of pine trees in response to 45° inclination at early times (1 h). Arabidopsis thaliana was stably transformed with a 35S::PrMADS11 construct in an effort to identify the putative targets of PrMADS11. A massive transcriptome analysis revealed 947 differentially expressed genes: 498 genes were up-regulated, and 449 genes were down-regulated due to the over-expression of PrMADS11. The gene ontology analysis highlighted a cell wall remodeling function among the differentially expressed genes, suggesting the active participation of cell wall modification required during the response to vertical stem loss. In addition, the phenylpropanoid pathway was also indicated as a PrMADS11 target, displaying a marked increment in the expression of the genes driven to the biosynthesis of monolignols. The EMSA assays confirmed that PrMADS11 interacts with CArG-box sequences. This TF modulates the gene expression of several molecular pathways, including other TFs, as well as the genes involved in cell wall remodeling. The increment in the lignin content and the genes involved in cell wall dynamics could be an indication of the key role of PrMADS11 in the response to trunk inclination.

Assuntos

Regulação da Expressão Gênica de Plantas , Pinus , Proteínas de Plantas , Pinus/genética , Pinus/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Caules de Planta/metabolismo , Caules de Planta/genética , Parede Celular/metabolismo , Parede Celular/genética , Perfilação da Expressão Gênica , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Lignina/metabolismo , Lignina/biossíntese , Proteínas de Domínio MADS/genética , Proteínas de Domínio MADS/metabolismo , Plantas Geneticamente Modificadas/genética

RESUMO

Despite the promising potential of Solanum plant glycoalkaloids in combating skin cancer, their clinical trials have been halted due to dose-dependent toxicity and poor water solubility. In this study, we present a rational approach to address these limitations and ensure colloidal stability of the nanoformulation over time by designing solid lipid-polymer hybrid nanoparticles (SLPH). Leveraging the biocompatible and cationic properties of polyaspartamides, we employed a new polyaspartamide derivative (P1) as a raw material for this class of nanostructures. Subsequently, we prepared SLPH through a one-step process involving hot-melt emulsification followed by ultrasonication. The physicochemical properties of the SLPH were thoroughly characterized using dynamic light scattering (DLS), ζ-potential analysis, nanoparticle tracking analysis (NTA), differential scanning calorimetry (DSC), Fourier-transform infrared spectroscopy (FT-IR), and transmission electron microscopy (TEM). The optimized formulation exhibited long-term stability over six months under low temperatures, maintaining a particle size around 200 nm, a polydispersity index (PdI) lower than 0.2, and a ζ-potential between +35-40 mV. Furthermore, we evaluated the cytotoxic effect of the SLPH against human cutaneous melanoma cells (SK-MEL-28) compared to human foreskin fibroblast cells (HFF-1). Encapsulation of glycoalkaloids into the nanoparticles (SLPH-GE) resulted in a two-fold greater selective cytotoxic profile for melanoma cells than glycoalkaloids-free (GE). The nanoparticles disrupted the stratum corneum barrier with a penetration depth of approximately 77 µm. These findings underscore the potential of the developed nanosystem as an effective glycoalkaloid carrier with suitable colloidal and biological properties for further studies in topical treatment strategies for cutaneous melanoma.

Assuntos

Lipídeos , Melanoma , Nanopartículas , Polímeros , Humanos , Nanopartículas/química , Lipídeos/química , Melanoma/tratamento farmacológico , Melanoma/patologia , Polímeros/química , Polímeros/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/química , Tamanho da Partícula , Alcaloides/química , Alcaloides/farmacologia , Linhagem Celular Tumoral , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/patologia , Antineoplásicos/farmacologia , Antineoplásicos/química , Administração Tópica , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Propriedades de Superfície

RESUMO

The neem tree (Azadirachta indica A. Juss) is grown mainly for shade, fuel, and numerous non-timber forest products using its leaves, fruit, and bark. It produces an essential oil that is used as a source for obtaining bioinsecticides, with a broad spectrum of action in agricultural production. Its bioinsecticidal activity is due to the presence of triterpenes, such as azadirachtin, a product in continued growth of the global biopesticide market. Optimal conditions for neem oil extraction using response surface methodology (RSM) and microwave-assisted extraction (MAE) methods have been defined. However, the extraction conditions for these methods tend to consume high volumes of organic solvent and long extraction times. The aim of the present study is to determine the optimal conditions for the extraction of azadirachtin from neem seeds in a hydroalcoholic medium using MAE and RSM with a Box-Behnken design (BBD). A BBD was applied to evaluate the effects of the factors, magnetron voltage (X1), extraction time (X2), and pH of the extraction medium (X3), on the yield of the azadirachtin extraction process. The effect of each variable on the extraction yield was studied independently, considering the pure coefficients (linear and quadratic) on the three levels that were studied in the experiments. Moreover, the study experiments were conducted in triplicate, data were presented as mean and standard deviation, homogeneity of variances was estimated using Levene's test, and a two-way ANOVA with Tukey's post hoc analysis was performed to identify the experimental conditions that allowed us to find the highest extraction yield and to analyze whether the response surface model adequately described our data. The most significant effects of the model correspond to quadratic and interaction effects (p < 0.0001); the quadratic terms voltage (X1), extraction time (X2), and pH (X3); and the interaction effects between voltage-pH (X1*X3) and time-pH (X2*X3), which had a significant influence on the model. Moreover, a canonical analysis was performed. The optimal conditions were as follows: 69.22 V, 6.89 min, and a pH value of 4.35, coinciding with the zones shown in the contour plots. Furthermore, the response obtained at the optimal conditions was 37.5 µg of azadirachtin per gram of pretreated seed.

RESUMO

The extraction of geraniol from palmarosa oil using hydrotropic solvents was investigated. Palmarosa oil possesses an appealing rose aroma and properties like anti - inflammatory, antifungal, and antioxidant due to the presence of geraniol. The extraction of geraniol from palmarosa oil by using distillation methods like steam dis tillation and fractional distillation was a laborious process. So hydrotropes were tried for extraction. The geraniol yield and purity depend on parameters like concentration of hydrotrope, solvent volume ratio, and time period. Using the Box Benkhem Desig n (BBD), the extraction process was optimized. One of the major advantages of using hydrotropic solvents is that they were classified as green solvents, and recovery of solvents is also possible. To reduce the extraction time probe sonication is carried ou t. Different hydrotropic solvents with probe sonication are done on palmarosa oil by altering various process parameters to study the separation, yield, and purity.

---

Se investigó la extracción de geraniol del aceite de palmarosa utilizando solventes hidrotrópicos. El aceite de palmarosa posee un atractivo aroma a rosa y propiedades antiinflamatorias, antifúngicas y antioxidantes debido a la pr esencia de geraniol. La extracción de geraniol del aceite de palmarosa mediante métodos de destilación como la destilación por vapor y la destilación fraccionada ha sido un proceso laborioso. Por lo tanto, se probaron los hidrotropos para la extracción. El rendimiento y la pureza del geraniol dependen de parámetros como la concentración del hidrotropo, la relación de volumen del solvente y el período de tiempo. Se optimizó el proceso de extracción usando el diseño Box Benkhem (BBD). Una de las principales v entajas de usar solventes hidrotrópicos es que se clasifican como solventes verdes y también es posible recuperar los solventes. Para reducir el tiempo de extracción, se lleva a cabo una sonda de ultrasonido. Se realizan diferentes solventes hidrotropos co n sonda de ultrasonido en el aceite de palmarosa alterando varios parámetros del proceso para estudiar la separación, el rendimiento y la pureza.

Assuntos

Óleos de Plantas/química , Cymbopogon/química , Monoterpenos Acíclicos/química , Cromatografia Gasosa

RESUMO

PURPOSE: For growth of methylotrophic yeast, glycerol is usually used as a carbon source. Glucose is used in some cases, but not widely consumed due to strong repressive effect on AOX1 promoter. However, glucose is still considered as a carbon source of choice since it has low production cost and guarantees growth rate comparable to glycerol. RESULTS: In flask cultivation of the recombinant yeast, Pichia pastoris GS115(pPIC9K-appA38M), while methanol induction point(OD600) and methanol concentration significantly affected the phytase expression, glucose addition in induction phase could enhance phytase expression. The optimal flask cultivation conditions illustrated by Response Surface Methodology were 10.37 OD600 induction point, 2.02 h before methanol feeding, 1.16% methanol concentration and 40.36µL glucose feeding amount(for 20 mL culture volume) in which the expressed phytase activity was 613.4 ± 10.2U/mL, the highest activity in flask cultivation. In bioreactor fermentation, the intermittent glucose feeding showed several advantageous results such as 68 h longer activity increment, 149.2% higher cell density and 200.1% higher activity compared to the sole methanol feeding method. These results implied that remaining glucose at induction point might exhibit a positive effect on the phytase expression. CONCLUSION: Glucose intermittent feeding could be exploited for economic phytase production and the other recombinant protein expression by P. pastoris GS115.

Assuntos

6-Fitase , Reatores Biológicos , Fermentação , Glucose , Metanol , Proteínas Recombinantes , 6-Fitase/genética , 6-Fitase/metabolismo , Glucose/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Metanol/metabolismo , Reatores Biológicos/microbiologia , Meios de Cultura/química , Meios de Cultura/metabolismo , Saccharomycetales/genética , Saccharomycetales/metabolismo , Saccharomycetales/crescimento & desenvolvimento , Pichia/genética , Pichia/metabolismo , Pichia/crescimento & desenvolvimento , Expressão Gênica

RESUMO

Chloraluminium phthalocyanine (ClAlPc) has potential therapeutic effect for the treatment of cancer; however, the molecule is lipophilic and may present self-aggregation which limits its clinical success. Thus, nanocarriers like liposomes can improve ClAlPc solubility, reduce off-site toxicity and increase circulation time. For this purpose, developing suitable liposomes requires the evaluation of different lipid compositions. Herein, we aimed to develop liposomes containing soy phosphatidylcholine (SPC), 1,2-distearoyl-sn-glycero- 3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPEPEG2000), cholesterol and oleic acid loaded with ClAlPc using the surface response methodology and the Box-Behnken design. Liposomes with particle size from 110.93 to 374.97 nm and PdI from 0.265 to 0.468 were obtained. The optimized formulation resulted in 69.09 % of ClAlPc encapsulated, with particle size and polydispersity index, respectively, at 153.20 nm and 0.309, providing stability and aggregation control. Atomic force microscopy revealed vesicles in a spherical or almost spherical shape, while the analyzes by Differential Scanning Calorimetry (DSC), Powder X-ray Diffraction (PXRD), and Fourier transform infrared spectroscopy (FTIR) suggested that the drug was adequately incorporated into the lipid bilayer of liposomes, in its amorphous state or molecularly dispersed. In vitro studies conducted in breast cancer cells (4T1) showed that liposome improved phototoxicity compared to the ClAlPc solution. ClAlPc-loaded liposomes also enhanced the production of ROS 3-fold compared to the ClAlPc solution. Finally, confocal microscopy and flow cytometry demonstrated the ability of the liposomes to enter cells and deliver the fluorescent ClAlPc photosensitizer with dose and time-dependent effects. Thus, this work showed that Box-Behnken factorial design was an effective strategy for optimizing formulation development. The obtained ClAlPc liposomes can be applied for photodynamic therapy in breast cancer cells.

Assuntos

Neoplasias da Mama , Indóis , Lipossomos , Compostos Organometálicos , Tamanho da Partícula , Fotoquimioterapia , Fármacos Fotossensibilizantes , Fotoquimioterapia/métodos , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Indóis/química , Indóis/administração & dosagem , Feminino , Compostos Organometálicos/química , Compostos Organometálicos/administração & dosagem , Humanos , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/administração & dosagem , Fármacos Fotossensibilizantes/farmacologia , Linhagem Celular Tumoral , Polietilenoglicóis/química , Fosfatidiletanolaminas/química , Fosfatidilcolinas/química , Colesterol/química , Ácido Oleico/química

RESUMO

Background: Orbital hypertelorism is a rare congenital condition caused by craniofacial malformations. It consists of complete orbital lateralization, characterized by an increase in distance (above the 95th percentile) of the inner canthal (ICD), outer canthal, and interpupillary distances. It can be approached surgically, and the main techniques are box osteotomy and facial bipartition. The surgical procedure is usually performed before the age of 8. We describe here two patients who underwent late surgical correction using the box osteotomy technique. Case Description: Patient 1: A 13-year-old female presenting isolated hypertelorism with 5 cm ICD and left eye amblyopia. Patient 2: A 15-year-old female with orbital hypertelorism, 4.6 cm ICD, and nasal deformity. Both patients underwent orbital translocation surgery and had no neurological disorders. Conclusion: The article reports two cases of isolated hypertelorism treated late with the box osteotomy technique. Both surgeries were successful, with no postoperative complications. It appears that it is possible to obtain good surgical results even in patients who have not been able to undergo surgery previously.

RESUMO

In recent years, antimony sulfide (Sb2S3) has been investigated as a photovoltaic absorber material due to its suitable absorber coefficient, direct band gap, extinction coefficient, earth-abundant, and environmentally friendly constituents. Therefore, this work proposes Sb2S3 film preparation by an effective two-step process using a new graphite box design and sulfur distribution, which has a high repeatability level and can be scalable. First, an Sb thin film was deposited using the RF-Sputtering technique, and after that, the samples were annealed with elemental sulfur into a graphite box, varying the sulfurization time from 20 to 50 min. The structural, optical, morphological, and chemical characteristics of the resulting thin films were analyzed. Results reveal the method's effectivity and the best properties were obtained for the sample sulfurized during 40 min. This Sb2S3 thin film presents an orthorhombic crystalline structure, elongated grains, a band gap of 1.69 eV, a crystallite size of 15.25 Å, and a nearly stoichiometric composition. In addition, the formation of a p-n junction was achieved by depositing silver back contact on the Glass/FTO/CdS/Sb2S3 structure. Therefore, the graphite box design has been demonstrated to be functional to obtain Sb2S3 by a two-step process.

RESUMO

Introduction: A Computer-Assisted Detection (CAD) System for classification into malignant-benign classes using CT images is proposed. Methods: Two methods that use the fractal dimension (FD) as a measure of the lung nodule contour irregularities (Box counting and Power spectrum) were implemented. The LIDC-IDRI database was used for this study. Of these, 100 slices belonging to 100 patients were analyzed with both methods. Results: The performance between both methods was similar with an accuracy higher than 90%. Little overlap was obtained between FD ranges for the different malignancy grades with both methods, being slightly better in Power spectrum. Box counting had one more false positive than Power spectrum. Discussion: Both methods are able to establish a boundary between the high and low malignancy degree. To further validate these results and enhance the performance of the CAD system, additional studies will be necessary.

RESUMO

The CACCC-box motif emerges as a pivotal cis-regulatory element implicated in diverse developmental processes and diseases, particularly cardiovascular diseases (CVDs). This study centers on the intricate interplay between the CACCC-box and its binding proteins such as: the Krüppel-Like Family (KLF) of transcription factors as primary effectors in the context of CVDs. Our analysis was through a bioinformatics approach, which revealed significant transcriptional activity among KLF subgroup 2, exhibiting the highest number of interactions focusing on the established roles: pluripotency, cancer, and cardiovascular development and diseases. Our analysis reveals KLF's interactions with GATA4, MEF2C, NKX2.5 and other ~90 potential genes that participate in the regulation of the hypertrophic environment (or CVDs' Environment). Also, the GO analysis showed that genes containing the motif CACCC were enriched for multiple CVDs; in combination with STRING analysis, these results pointed to a link between KLFs and these diseases. The analysis further identifies other potential CACCC-box binding factors, such as SP family members, WT1, VEZF1, and -SALL4, which are implicated in cardiac contraction, remodeling, and inflammation processes.

RESUMO

In this study, a CoO-Fe2O3/SiO2/TiO2 (CIST) nanocomposite was synthesized and utilized as an adsorbent to remove methylene blue (MB), malachite green (MG), and copper (Cu) from aqueous environments. The synthesized nanocomposite was characterized using field emission scanning electron microscopy (FE-SEM), Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and X-ray diffraction (XRD). Input parameters included pH (3-10), contact time (10-30 min), adsorbent amount (0.01-0.03 g), and pollutant concentration (20-60 mg L-1). The effects of these parameters on the removal process efficiency were modeled and optimized using the response surface methodology (RSM) based on the Box-Behnken design (BBD). The RSM-BBD method demonstrated the capability to develop a second-degree polynomial model with high validity (R2 ˃ 0.99) for the removal process. The optimization results using the RSM-BBD method revealed a removal efficiency of 98.01%, 93.06%, and 88.26% for MB, MG, and Cu, respectively, under optimal conditions. These conditions were a pH of 6, contact time of 10 min, adsorbent amount of 0.025 g, and concentration of 20 mg L-1. The synthesized adsorbent was recovered through five consecutive adsorption-desorption cycles using hydrochloric acid. The results showed an approximately 12% reduction from the first to the seventh cycle. Also, MB, MG, and Cu removal from real water samples in optimal conditions was achieved in the range of 81.69-98.18%. This study demonstrates the potential use of CIST nanocomposite as an accessible and reusable option for removing MB, MG, and Cu pollutants from aquatic environments.

RESUMO

Pectin is widely used in several products in the industry. Conventionally, strong and harmful acids are used for its extraction. This study optimized the extraction of orange peel's pectin using citric acid, considering yield and degree of esterification (DE) as response variables. Proximal analyses were performed, and the samples were subjected to a Box-Behnken design on three central points, considering as variables the temperature, time, and pH. The results of proximate analyses of the orange peels revealed 11.76 % moisture content, 87.26 % volatiles, 0.09 % ash, 50.45 % soluble carbohydrates, 70.60 % total carbohydrates, 0.89 % fixed carbon, 5.35 % lipids, and 36.75 mg GAE/g of phenolic compounds. The resulting second-order polynomial model described the relation of the input and output variables related to each other. The best performance to obtain a higher yield (18.18 %) of high methoxyl pectin (DE 50 %) was set at 100 °C/30 min/pH 2.48. Pectin showed antioxidant properties by ABTS and DPPH assays and similar thermal properties to the commercial polymer. Its equivalent weight was 1219.51 mol/g, and the methoxyl and anhydrouronic acid were 2.23 and 67.10 %, respectively. Hence, pectin extraction with citric acid results in a high-quality polymer and could be used as a gelling agent, stabilizer, or texturizer in food products.

Assuntos

Citrus sinensis , Pectinas , Pectinas/química , Citrus sinensis/química , Ácido Cítrico/química , Temperatura , Antioxidantes/farmacologia , Excipientes

RESUMO

Today, safeguarding sensitive content through encryption is crucial. This work presents a hybrid cryptosystem for images that employs both asymmetric and symmetric encryption. The asymmetric component involves applying the Diffie-Hellman protocol and the ElGamal cryptosystem to securely transmit two constants. These constants are necessary for the symmetrical aspect to generate dynamic permutations, substitution boxes, and round keys. Following an encryption process with fourteen rounds, the encrypted images are processed by an algorithm proposed to enhance entropy, a critical metric for assessing encryption quality. It increases the frequencies of the basic colors to achieve a histogram closely resembling a uniform distribution, but it increases the image size by approximately 8%. This improves the entropy values achieved by the hybrid cryptosystem, bringing them remarkably close to the ideal value of 8.0. In specific instances, the entropy values were elevated from 7.99926 to 8.0. The proposed method exhibits resilience against various attacks, including differential, linear, brute force, and algebraic attacks, as evaluated through the entropy, correlation, goodness of fit, Discrete Fourier Transform (DFT), Number of Pixels Change Rate (NPCR), Unified Average Changing Intensity (UACI), Avalanche Criteria (AC), contrast, energy, and homogeneity. Further, encrypted images are subjected to noise attacks ranging from 20% to 50% noise, including additive, multiplicative, occlusion noise, as well as the newly introduced χ2 noise. The noise damage is quantified using the proposed Similarity Parameter (SP), and a 3 × 3 median filter is employed to enhance the visual quality.

RESUMO

KEY MESSAGE: Lastly, the bZIP gene family encompasses genes that have been reported to play a role in flower development, such as bZIP14 (FD). Notably, bZIP14 is essential for Flowering Locus T (FT) initiation of floral development in Arabidopsis (Abe et al. 2005). Cotton (Gossypium hirsutum L.) is the world's most extensively cultivated fiber crop. However, its reproductive development is poorly characterized at the molecular level. Thus, this study presents a detailed transcriptomic analysis of G. hirsutum at three different reproductive stages. We provide evidence that more than 64,000 genes are active in G. hirsutum during flower development, among which 94.33% have been assigned to functional terms and specific pathways. Gene set enrichment analysis (GSEA) revealed that the biological process categories of floral organ development, pollen exine formation, and stamen development were enriched among the genes expressed during the floral development of G. hirsutum. Furthermore, we identified putative Arabidopsis homologs involved in the G. hirsutum gene regulatory network (GRN) of pollen and flower development, including transcription factors such as WUSCHEL (WUS), INNER NO OUTER (INO), AGAMOUS-LIKE 66 (AGL66), SPOROCYTELESS/NOZZLE (SPL/NZZ), DYSFUNCTIONAL TAPETUM 1 (DYT1), ABORTED MICROSPORES (AMS), and ASH1-RELATED 3 (ASHR3), which are known crucial genes for plant reproductive success. The cotton MADS-box protein-protein interaction pattern resembles the previously described patterns for AGAMOUS (AG), SEEDSTICK (STK), SHATTERPROOF (SHP), and SEPALLATA3 (SEP3) homolog proteins from Arabidopsis. In addition to serving as a resource for comparative flower development studies, this work highlights the changes in gene expression profiles and molecular networks underlying stages that are valuable for cotton breeding improvement.

Assuntos

Flores , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , Gossypium , Gossypium/genética , Gossypium/crescimento & desenvolvimento , Gossypium/fisiologia , Flores/genética , Flores/crescimento & desenvolvimento , Reprodução/genética , Transcriptoma , Perfilação da Expressão Gênica , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/fisiologia

RESUMO

DEAD-box helicases are global regulators of liquid-liquid phase separation (LLPS), a process that assembles membraneless organelles inside cells. An outstanding member of the DEAD-box family is DDX3X, a multi-functional protein that plays critical roles in RNA metabolism, including RNA transcription, splicing, nucleocytoplasmic export, and translation. The diverse functions of DDX3X result from its ability to bind and remodel RNA in an ATP-dependent manner. This capacity enables the protein to act as an RNA chaperone and an RNA helicase, regulating ribonucleoprotein complex assembly. DDX3X and its orthologs from mouse, yeast (Ded1), and C. elegans (LAF-1) can undergo LLPS, driving the formation of neuronal granules, stress granules, processing bodies or P-granules. DDX3X has been related to several human conditions, including neurodevelopmental disorders, such as intellectual disability and autism spectrum disorder. Although the research into the pathogenesis of aberrant biomolecular condensation in neurodegenerative diseases is increasing rapidly, the role of LLPS in neurodevelopmental disorders is underexplored. This review summarizes current findings relevant for DDX3X phase separation in neurodevelopment and examines how disturbances in the LLPS process can be related to neurodevelopmental disorders.

Assuntos

Transtorno do Espectro Autista , RNA Helicases DEAD-box , Transtornos do Neurodesenvolvimento , Animais , Humanos , Camundongos , Transtorno do Espectro Autista/genética , Caenorhabditis elegans/metabolismo , RNA Helicases DEAD-box/genética , RNA Helicases DEAD-box/metabolismo , Transtornos do Neurodesenvolvimento/genética , RNA/metabolismo , Saccharomyces cerevisiae/metabolismo

RESUMO

Frog virus 3 (FV3) and related ranaviruses are emerging infectious disease threats to ectothermic vertebrate species globally. Although the impact of these viruses on amphibian health is relatively well studied, less is understood about their effects on reptile health. We report two cases of FV3 infection, 11 mo apart, in three-toed box turtles (Terrapene mexicana triunguis) from a wildlife rehabilitation center. Case 1 had upper respiratory signs upon intake but had no clinical signs at the time of euthanasia 1 mo later. Case 2 presented for vehicular trauma, had ulcerative pharyngitis and glossitis, and died overnight. In case 1, we detected FV3 nucleic acid with qPCR in oral swabs, kidney, liver, spleen, and tongue. In case 2, we detected FV3 in an oral swab, an oral plaque, heart, kidney, lung, liver, spleen, and tongue. We also detected FV3 nucleic acid with in situ hybridization for case 2. For both cases, FV3 was isolated in cell culture and identified with DNA sequencing. Histopathologic examination of postmortem tissue from case 1 was unremarkable, whereas acute hemorrhagic pneumonia and splenic necrosis were noted in case 2. The difference in clinical signs between the two cases may have been due to differences in the temporal course of FV3 disease at the time of necropsy. Failure to detect this infection previously in Missouri reptiles may be due to lack of surveillance, although cases may also represent a novel spillover to box turtles in Missouri. Our findings reiterate previous suggestions that the range of FV3 infection may be greater than previously documented and that infection may occur in host species yet to be tested.

Assuntos

Infecções por Vírus de DNA , Ácidos Nucleicos , Ranavirus , Tartarugas , Animais , Missouri/epidemiologia , Animais Selvagens , Infecções por Vírus de DNA/veterinária

RESUMO

ABSTRACTObjetives: Omega-3 (n3) fatty acids have been studied as an option to alleviate the harmful effects of obesity. However, its role in obesity-related behavioral changes is still controversial. This study aimed to evaluate the effects of n3 on behavior and neuroinflammation in obese animals. Methods: Male Wistar rats were divided into four groups: control diet (CT), CT+n3, cafeteria diet (CAF), and CAF+n3. Diet was administered for 13 weeks, and n3 was supplemented during the last 5 weeks. Metabolic and biochemical parameters were evaluated, as well as anxiety-like behaviors. Immunoblots were conducted in the animals' cerebral cortex and hippocampus to assess changes in neuroinflammatory markers.Results: CAF-fed animals showed higher weight gain, visceral adiposity, fasting glucose, total cholesterol, triglycerides, and insulin levels, and n3 improved the lipid profile and restored insulin sensitivity. CAF-fed rats showed anxiety-like behaviors in the open field and light-dark box tasks but not in the contextual aversive conditioning. Omega-3 did not exert any effect on these behaviors. Regarding neuroinflammation, diet and supplementation acted in a region-specific manner. In the hippocampus, CAF reduced claudin-5 expression with no effect of n3, indicating a brain-blood barrier disruption following CAF. Furthermore, in the hippocampus, the glial fibrillary acidic protein (GFAP) and toll-like receptor 4 (TLR-4) were reduced in treated obese animals. However, n3 could not reverse the TLR-4 expression increase in the cerebral cortex.Discussion: Although n3 may protect against some neuroinflammatory manifestations in the hippocampus, it does not seem sufficient to reverse the increase in anxiolytic manifestations caused by CAF.

Assuntos

Ácidos Graxos Ômega-3 , Receptor 4 Toll-Like , Ratos , Masculino , Animais , Ratos Wistar , Doenças Neuroinflamatórias , Obesidade/etiologia , Obesidade/metabolismo , Dieta , Ácidos Graxos Ômega-3/farmacologia , Ansiedade/etiologia , Ansiedade/prevenção & controle , Suplementos Nutricionais

RESUMO

Los deportes de combate conllevan un alto riesgo de padecer traumatismos dentoalveolares. Este estudio se evaluó la incidencia de traumatismos y lesiones orofaciales relacionadas con los deportes de combate, junto con la evaluación de las actitudes y hábitos de los deportistas respecto al uso de protectores bucales. Este estudio se realizó mediante sistema de encuestas digitales distribuidas en diferentes clubes deportivos donde se practican deportes de combate y no combate. Los deportistas de combate tienen más probabilidad sufrir alguna lesión orofacial que los no deportistas de combate. El sexo masculino y la experiencia son factores predisponentes en la aparición de traumatismos y lesiones orofaciales, siendo las laceraciones de tejidos blandos las más frecuentes. Los deportistas de combate llevan más protectores bucales que los deportistas en general, siendo el protector tipo II el más usado. Los deportistas que no llevan protectores no consideran que sea necesario. En caso de sufrir una avulsión dental, la mayoría de los participantes consideran que es posible reimplantar un diente avulsionado.

---

Combat sports carry a high risk of suffering dentoalveolar trauma. This study evaluated the incidence of dentoalveolar lesions related to combat sports, together with the evaluation of athletes' attitudes and habits regarding the use of mouth guards. This study was carried out using a digital survey system distributed in different sports clubs where combat and non-combat sports are practiced. Combat athletes are more likely to suffer a dentoalveolar lesion than non-combat athletes. Male sex and experience are predisposing factors in the appearance of dentoalveolar lesions, with soft tissue lacerations being the most frequent. Combat athletes wear more mouth guards than athletes in general, with the type II protector being the most used. Athletes who do not wear protectors do not consider it necessary. In case of suffering a dental avulsion, most of the participants consider that it is possible to reimplant an avulsed tooth.