Three-Dimensional Cell Cultures as a Research Platform in Lung Diseases and COVID-19

da Silva da Costa Felipe Allan,Soares Murilo Racy,Malagutti-Ferreira Maria José,da Silva Gustavo Ratti,Lívero Francislaine Aparecida dos Reis,Ribeiro-Paes João Tadeu 한국조직공학과 재생의학회 2021 조직공학과 재생의학 Vol.18 No.5

Background: Chronic respiratory diseases (CRD) are a major public health problem worldwide. In the current epidemiological context, CRD have received much interest when considering their correlation with greater susceptibility to SARS-Cov-2 and severe disease (COVID-19). Increasingly more studies have investigated pathophysiological interactions between CRD and COVID-19. Area covered: Animal experimentation has decisively contributed to advancing our knowledge of CRD. Considering the increase in ethical restrictions in animal experimentation, researchers must focus on new experimental alternatives. Two-dimensional (2D) cell cultures have complemented animal models and significantly contributed to advancing research in the life sciences. However, 2D cell cultures have several limitations in studies of cellular interactions. Three-dimensional (3D) cell cultures represent a new and robust platform for studying complex biological processes and are a promising alternative in regenerative and translational medicine. Expert opinion: Three-dimensional cell cultures are obtained by combining several types of cells in integrated and self-organized systems in a 3D structure. These 3D cell culture systems represent an efficient methodological approach in studies of pathophysiology and lung therapy. More recently, complex 3D culture systems, such as lung-on-a-chip, seek to mimic the physiology of a lung in vivo through a microsystem that simulates alveolar-capillary interactions and exposure to air. The present review introduces and discusses 3D lung cultures as robust platforms for studies of the pathophysiology of CRD and COVID-19 and the mechanisms that underlie interactions between CRD and COVID-19. Background: Chronic respiratory diseases (CRD) are a major public health problem worldwide. In the current epidemiological context, CRD have received much interest when considering their correlation with greater susceptibility to SARS-Cov-2 and severe disease (COVID-19). Increasingly more studies have investigated pathophysiological interactions between CRD and COVID-19. Area covered: Animal experimentation has decisively contributed to advancing our knowledge of CRD. Considering the increase in ethical restrictions in animal experimentation, researchers must focus on new experimental alternatives. Two-dimensional (2D) cell cultures have complemented animal models and significantly contributed to advancing research in the life sciences. However, 2D cell cultures have several limitations in studies of cellular interactions. Three-dimensional (3D) cell cultures represent a new and robust platform for studying complex biological processes and are a promising alternative in regenerative and translational medicine. Expert opinion: Three-dimensional cell cultures are obtained by combining several types of cells in integrated and self-organized systems in a 3D structure. These 3D cell culture systems represent an efficient methodological approach in studies of pathophysiology and lung therapy. More recently, complex 3D culture systems, such as lung-on-a-chip, seek to mimic the physiology of a lung in vivo through a microsystem that simulates alveolar-capillary interactions and exposure to air. The present review introduces and discusses 3D lung cultures as robust platforms for studies of the pathophysiology of CRD and COVID-19 and the mechanisms that underlie interactions between CRD and COVID-19.

Saline extract of Portulaca elatior leaves with photoprotective and antioxidant activities does not show acute oral and dermal toxicity in mice

da Silva Suéllen Pedrosa,da Costa Clarice Barbosa Lucena,de Freitas Anderson Felipe Soares,da Silva José Dayvid Ferreira,Costa Wêndeo Kennedy,da Silva Wênio Sandoval Filho Lima,Machado Janaina Carla B 한국독성학회 2023 Toxicological Research Vol.39 No.2

The present study aimed to evaluate saline extracts from the leaves (LE) and stem (SE) of Portulaca elatior in relation to their phytochemical composition and photoprotective and antioxidant effects, as well as to evaluate the toxicity of the leaf extract. The extracts were characterized for protein concentration and phenol and flavonoid contents, as well as for thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) profiles. Total antioxidant capacity and DPPH and ABTS+ scavenging activities were determined. In the photoprotective activity assay, the sun protection factor (SPF) was calculated. The toxicity evaluation of LE included in vitro hemolytic assay and in vivo oral and dermal acute toxicity assays in Swiss mice. LE showed the highest protein, phenol, and flavonoid (8.79 mg/mL, 323.46 mg GAE/g, and 101.96 QE/g, respectively). TLC revealed the presence of flavonoids, reducing sugars, terpenes, and steroids in both extracts. In HPLC profiles, LE contained flavonoids, while SE contained flavonoids and ellagic tannins. The antioxidant activity assays showed the lowest IC50 values (34.15–413.3 μg/mL) for LE, which presented relevant SPF (> 6) at 50 and 100 μg/mL. LE demonstrated low hemolytic capacity, and no signs of intoxication were observed in mice treated orally or topically at 1000 mg/kg. However, at 2000 mg/kg, an increase in the mean corpuscular volume of erythrocytes and a reduction in lymphocytes were observed; animals treated topically with 2000 mg/kg displayed scratching behavior during the first hour of observation and showed edema and erythema that regressed after six days. In conclusion, LE did not present acute oral or dermal toxicity in Swiss mice at a dose of 1000 mg/kg and showed slight toxicity in animals treated with 2000 mg/kg.

Track and Field Practice and Bone Outcomes among Adolescents: A Pilot Study (ABCD-Growth Study)

Yuri da Silva Ventura Faustino-da-Silva,Ricardo Ribeiro Agostinete,André Oliveira Werneck,Santiago Maillane-Vanegas,Kyle Robinson Lynch,Isabella Neto Exupério,Igor Hideki Ito,Romulo Araújo Fernandes 대한골대사학회 2018 대한골대사학회지 Vol.25 No.1

Background: Osteoporosis is considered a public health problem with high worldwide prevalence. One approach to prevention is through the promotion of physical activity, especially exercise, during adolescence. Methods: This study compared bone variables in different body segments in adolescents according to participation in track and field. The study included 34 adolescents (22 boys), of whom 17 were track and field athletes and 17 were control subjects. Bone mineral density (BMD, g/cm2) and bone mineral content (BMC, g) were analyzed using dual energy X-ray absorptiometry (total body stratified by body segments). Peak height velocity was used to estimate somatic maturation. Results: Athletes had higher BMD (P=0.003) and BMC (P=0.011) values in the lower limbs and higher whole body BMD (P=0.025) than the control group. However, when adjusted for confounding factors, the difference was not maintained. The groups had similar lean soft tissue values (P=0.094). Training overload was positively correlated with BMD in the upper limbs (r=0.504; 95% confidence interval, 0.031-0.793). Although track and field athletes had higher BMD and BMC values in the lower limbs, these differences were not significant when adjusted for confounding factors. Conclusions: Track and field participation in adolescence appears to influence BMD and BMC in lower limbs, and fat-free mass seems to mediate this effect. Also, higher training loads were found to be positive for bone health in upper limbs.

Cone-beam computed tomography findings of impacted upper canines

Da Silva Santos, Ludmilla Mota,Bastos, Luana Costa,Oliveira-Santos, Christiano,Da Silva, Silvio Jose Albergaria,Neves, Frederico Sampaio,Campos, Paulo Sergio Flores Korean Academy of Oral and Maxillofacial Radiology 2014 Imaging Science in Dentistry Vol.44 No.4

Purpose: To describe the features of impacted upper canines and their relationship with adjacent structures through three-dimensional cone-beam computed tomography (CBCT) images. Materials and Methods: Using the CBCT scans of 79 upper impacted canines, we evaluated the following parameters: gender, unilateral/bilateral occurrence, location, presence and degree of root resorption of adjacent teeth (mild, moderate, or severe), root dilaceration, dental follicle width, and presence of other associated local conditions. Results: Most of the impacted canines were observed in females (56 cases), unilaterally (51 cases), and at a palatine location (53 cases). Root resorption in adjacent teeth and root dilaceration were observed in 55 and 47 impacted canines, respectively. In most of the cases, the width of the dental follicle of the canine was normal; it was abnormally wide in 20 cases. A statistically significant association was observed for all variables, except for root dilaceration (p=0.115) and the side of impaction (p=0.260). Conclusion: Root resorption of adjacent teeth was present in most cases of canine impaction, mostly affecting adjacent lateral incisors to a mild degree. A wide dental follicle of impacted canines was not associated with a higher incidence of external root resorption of adjacent teeth.

Application of Arbuscular Mycorrhizal Fungi during the Acclimatization of Alpinia purpurata to Induce Tolerance to Meloidogyne arenaria

da Silva Campos, Maryluce Albuquerque,da Silva, Fabio Sergio Barbosa,Yano-Melo, Adriana Mayumi,de Melo, Natoniel Franklin,Maia, Leonor Costa The Korean Society of Plant Pathology 2017 Plant Pathology Journal Vol.33 No.3

An experiment was conducted to evaluate the tolerance of micropropagated and mycorrhized alpinia plants to the parasite Meloidogyne arenaria. The experimental design was completely randomized with a factorial arrangement of four inoculation treatments with arbuscular mycorrhizal fungi (AMF) (Gigaspora albida, Claroideoglomus etunicatum, Acaulospora longula, and a non-inoculated control) in the presence or absence of M. arenaria with five replicates. The following characteristics were evaluated after 270 days of mycorrhization and 170 days of M. arenaria inoculation: height, number of leaves and tillers, fresh mass of aerial and subterranean parts, dry mass of aerial parts, foliar area, nutritional content, mycorrhizal colonization, AMF sporulation, and the number of galls, egg masses, and eggs. The results indicated a significant interaction between the treatments for AMF spore density, total mycorrhizal colonization, and nutrient content (Zn, Na, and N), while the remaining parameters were influenced by either AMF or nematodes. Plants inoculated with A. longula or C. etunicatum exhibited greater growth than the control. Lower N content was observed in plants inoculated with AMF, while Zn and Na were found in larger quantities in plants inoculated with C. etunicatum. Fewer galls were observed on mycorrhized plants, and egg mass production and the number of eggs were lower in plants inoculated with G. albida. Plants inoculated with A. longula showed a higher percentage of total mycorrhizal colonization in the presence of the nematode. Therefore, the association of micropropagated alpinia plants and A. longula enhanced tolerance to parasitism by M. arenaria.

Reaction of Cauliflower Genotypes to Black Rot of Crucifers

da Silva, Lincon Rafael,da Silva, Renan Cesar Dias,Cardoso, Atalita Francis,de Mello Pela, Glaucia,Carvalho, Daniel Diego Costa The Korean Society of Plant Pathology 2015 Plant Pathology Journal Vol.31 No.2

This study aimed to evaluate six cauliflower genotypes regarding their resistance to black rot and their production performance. To do so, it was conducted two field experiments in Ipameri, $Goi\acute{a}s$, Brazil, in 2012 and 2013. It was used a randomized block design, with four replications (total of 24 plots). Each plot consisted of three planting lines 2.5 m long (six plants/line), spaced 1.0 m apart, for a total area of $7.5m^2$. Evaluations of black rot severity were performed at 45 days after transplanting, this is, 75 days after sowing (DAS), and yield evaluations at 90 to 105 DAS. The Verona 184 genotype was the most resistant to black rot, showing 1.87 and 2.25% of leaf area covered by black rot symptom (LACBRS) in 2012 and 2013. However, it was not among the most productive materials. The yield of the genotypes varied between 15.14 and 25.83 t/ha in both years, Lisvera F1 (21.78 and 24.60 t/ha) and Cindy (19.95 and 23.56 t/ha) being the most productive. However, Lisvera F1 showed 6.37 and 9.37% of LACBRS and Cindy showed 14.25 and 14.87% of LACBRS in 2012 and 2013, being both considered as tolerant to black rot.

Evaluation of salt and root-zone temperature stresses in leafy vegetables using hydroponics as a clean production cultivation technique in northeastern Brazil

da Silva Mairton Gomes,Gheyi Hans Raj,da Silva Lucas Lesqueves,de Souza Thaisa Teixeira,Silva Petterson Costa Conceição,Queiroz Laila de Andrade,dos Santos Thainan Sipriano,Soares Tales Miler 한국원예학회 2024 Horticulture, Environment, and Biotechnology Vol.65 No.1

Hydroponics has represented an emerging solution for regions with freshwater scarcity. Besides requiring a lower volume of water for production, this cultivation technique allows the use of brackish waters with high salinity levels. Under normal cultivation conditions (i.e., without any environmental control), plants are exposed to other abiotic stresses, such as tem perature, which can further enhance the salinity eff ect. Given the above, three experiments were conducted using a nutrient f i lm technique (NFT) hydroponic system: from April to May (autumn) with endive ( Cichorium endivia L., cv. Gigante Barbarella), from May to July (autumn/winter) with lettuce ( Lactuca sativa L., cv. Rubinela and cv. Veneranda), and from August to October (winter/spring) with chicory ( Cichorium intybus L., cv. Pão de Açúcar and cv. Radiche Todo Ano), in 2021. The trials aimed to evaluate the isolated and/or combined eff ect of salt and root-zone temperature stresses on yield, water use effi ciency, and visual quality of these vegetables. In all experiments, plants were subjected to stresses for 25 days in a randomized block design with a 2 × 4 factorial arrangement: two levels of electrical conductivity of water –ECw (0.30 and 5.50 dS m –1 for endive and chicory; 0.25 and 6.50 dS m −1 for lettuce) combined with four root-zone temperatures – RZTs (ambient-ARZT, ARZT + 2 °C, ARZT + 4 °C, and ARZT + 6 °C, corresponding to the mean temperatures of 24.72, 26.90, 28.77, and 30.73 °C for endive; 24.20, 26.00, 28.00, and 29.99 °C for lettuce; 27.12, 28.93, 31.01, and 32.86 °C for chicory). The isolated eff ects of RZT stresses or ECw levels on endive traits were observed, with leaf fresh matter losses of approxi mately 17 and 44% at higher ARZT + 6 °C (mean of 30.73 °C) and salinity (ECw 5.50 dS m –1 ), respectively. Shoot fresh matter losses of around 32 and 52% (Veneranda and Rubinela lettuces, respectively) and 65% (Radiche Todo Ano chicory) occurred only due to salt stress. The interaction aff ected the shoot fresh matter of Pão de Açúcar chicory, with approximate losses of 80% at higher ARZT + 6 °C (mean of 32.86 °C) and salt stress (ECw 5.50 dS m –1 ) than ARZT and ARZT + 2 °C and without salt stress (ECw 0.30 dS m –1 ). Despite these reductions, visual quality of plants was not depreciated. Therefore, the present study suggested cultivating leafy vegetables such as lettuce, chicory, and endive using brackish waters, even under high temperatures of nutrient solutions.

Fractal dimension analysis as an easy computational approach to improve breast cancer histopathological diagnosis

Lucas Glaucio da Silva,Waleska Rayanne Sizinia da Silva Monteiro,Tiago Medeiros de Aguiar Moreira,Maria Aparecida Esteves Rabelo,Emílio Augusto Campos Pereira de Assis,de Souza Gustavo Torres 한국현미경학회 2021 Applied microscopy Vol.51 No.1

Histopathology is a well-established standard diagnosis employed for the majority of malignancies, including breast cancer. Nevertheless, despite training and standardization, it is considered operator-dependent and errors are still a concern. Fractal dimension analysis is a computational image processing technique that allows assessing the degree of complexity in patterns. We aimed here at providing a robust and easily attainable method for introducing computer-assisted techniques to histopathology laboratories. Slides from two databases were used: A) Breast Cancer Histopathological; and B) Grand Challenge on Breast Cancer Histology. Set A contained 2480 images from 24 patients with benign alterations, and 5429 images from 58 patients with breast cancer. Set B comprised 100 images of each type: normal tissue, benign alterations, in situ carcinoma, and invasive carcinoma. All images were analyzed with the FracLac algorithm in the ImageJ computational environment to yield the box count fractal dimension (Db) results. Images on set A on 40x magnification were statistically different ( p = 0.0003), whereas images on 400x did not present differences in their means. On set B, the mean Db values presented promissing statistical differences when comparing. Normal and/or benign images to in situ and/or invasive carcinoma (all p < 0.0001). Interestingly, there was no difference when comparing normal tissue to benign alterations. These data corroborate with previous work in which fractal analysis allowed differentiating malignancies. Computer-aided diagnosis algorithms may beneficiate from using Db data; specific Db cut-off values may yield ~ 99% specificity in diagnosing breast cancer. Furthermore, the fact that it allows assessing tissue complexity, this tool may be used to understand the progression of the histological alterations in cancer.

Effects of processing, particle size and moisturizing of sorghum-based feeds on pellet quality and broiler production

Patricia Garcia da Silva,Luana Martins Schaly Oliveira,Nayanne Rodrigues de Oliveira,Fabio Ataides de Moura Junior,Maura Regina Sousa Silva,Deibity Alves Cordeiro,Cibele Silva Minafra,Fabiana Ramos do 아세아·태평양축산학회 2018 Animal Bioscience Vol.31 No.1

Objective: This study aimed to assess the effect of pelleted and expanded sorghum-based feeds prepared with different moisture levels and particle size of ingredients on metabolizable energy, ileal digestibility of amino acids and broiler performance. Methods: The experiment was performed with 720 male broiler chicks of the Cobb strain, with treatments of six replications, with 15 birds each; they were arranged in a completely randomized design and 2×2×2 factorial scheme (pelleted or expanded feed processing, 0.8% or 1.6% moisture addition in the mixer, and particle size of 650 or 850 microns). Results: Higher pellet quality (pellets, % and pellet durability index [PDI]) was obtained in expanded diets and inclusion of 1.6% moisture. The particle size of 850 microns increased the PDI of final diet. All studied treatments had no significant effect on weight gain and broiler carcass and cut yields. Lower feed conversion occurred for birds fed pelleted feed at 42 d. The highest apparent metabolizable energy (AME) and apparent metabolizable energy corrected to zero nitrogen balance (AMEn) values of feed in the initial rearing phase (10 to 13 days) were observed in birds fed pelleted feed or for feed prepared with 1.6% moisture. The highest ileal digestibility coefficients of amino acids were obtained with the consumption of pelleted feed prepared with a particle size of 650 microns and 1.6% moisture. Conclusion: Pelleted feed prepared with a milling particle size of 650 microns and 1.6% moisture provided increased ileal digestibility of amino acids and AMEn in the starter period. However, the expanded feed improved pellet quality and feed conversion of broilers at 42 days of age. We conclude that factors such as moisture, particle size and processing affect the pellet quality, and therefore should be considered when attempting to optimize broiler performance.

New Insights in Purinergic Therapy: Novel Antagonists for Uridine 5′-Triphosphate-Activated P2Y Receptors from Brazilian Flora

Natiele Carla da Silva Ferreira,Roˆmulo Jose´ Soares-Bezerra,Rebeca Ferreira Couto da Silveira,Clayton Menezes da Silva,Carla Santos de Oliveira,Andrea Surrage Calheiros,Taˆnia Maria Alves,Carlos Leom 한국식품영양과학회 2019 Journal of medicinal food Vol.22 No.2

P2Y2 and P2Y4 receptors are physiologically activated by uridine 5′-triphosphate (UTP) and are widely expressed in many cell types in humans. P2Y2 plays an important role in inflammation and proliferation of tumor cells, which could be attenuated with the use of antagonists. However, little is known about the physiological functions related to P2Y4, due to the lack of selective ligands for these receptors. This can be solved through the search for novel compounds with antagonistic activity. The aim of this study was to discover new potential antagonist candidates for P2Y2 and P2Y4 receptors from natural products. We applied a calcium measurement methodology to identify new antagonist candidates for these receptors. First, we established optimal conditions for the calcium assay using J774.G8, a murine macrophage cell line, which expresses functional P2Y2 and P2Y4 receptors and then, we performed the screening of plant extracts at a cutoff concentration of 50 μg/mL. ATP and ionomycin, known intracellular calcium inductors, were used to stimulate cells. The calculated EC50 were 11 μM and 103 nM, respectively. These cells also responded to the UTP stimulation with an EC50 of 1.021 μM. Screening assays were performed and a total of 100 extracts from Brazilian plants were tested. Joannesia princeps Vell. (stem) and Peixotoa A. Juss (flower and leaf) extracts stood out due to their ability to inhibit UTP-induced responses without causing cytotoxicity, and presented an IC50 of 32.32, 14.99, and 12.98 μg/mL, respectively. Collectively, our results point to the discovery of potential antagonist candidates from Brazilian flora for UTP-activated receptors.