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Melanoma is a highly metastatic and rapidly progressing cancer, a leading cause of mortality among skin cancers. The melanoma microenvironment, formed from the activity of malignant cells on the extracellular matrix and the recruitment of immune cells, plays an active role in the development of drug resistance and tumor recurrence, which are clinical challenges in cancer treatment. These tumoral metabolic processes are affected by proteins, including Galectin-3 (Gal-3), which is extensively involved in cancer development. Previously, we characterized a partially methylated mannogalactan (MG-Pe) with antimelanoma activities. In vivo models of melanoma were used to observe MG-Pe effects in survival, spontaneous, and experimental metastases and in tissue oxidative stress. Analytical assays for the molecular interaction of MG-Pe and Gal-3 were performed using a quartz crystal microbalance, atomic force microscopy, and contact angle tensiometer. MG-Pe exhibits an additive effect when administered together with the chemotherapeutic agent dacarbazine, leading to increased survival of treated mice, metastases reduction, and the modulation of oxidative stress. MG-Pe binds to galectin-3. Furthermore, MG-Pe antitumor effects were substantially reduced in Gal-3/KO mice. Our results showed that the novel Gal-3 ligand, MG-Pe, has both antitumor and antimetastatic effects, alone or in combination with chemotherapy.

Melanoma is a highly metastatic and rapidly progressing cancer, a leading cause of mortality among skin cancers. The melanoma microenvironment, formed from the activity of malignant cells on the extracellular matrix and the recruitment of immune cells, plays an active role in the development of drug resistance and tumor recurrence, which are clinical challenges in cancer treatment. These tumoral metabolic processes are affected by proteins, including Galectin-3 (Gal-3), which is extensively involved in cancer development. Previously, we characterized a partially methylated mannogalactan (MG

Melanoma is a form of skin cancer with high mortality owing to its fast progression and metastatic capacity. The treatments available nowadays are only palliative in advanced stages of the disease. Thus, alternative therapies for cancer treatment are in demand, and molecules from natural sources, such as polysaccharides, could represent new possible therapeutic approaches. Polysaccharides of freshwater and marine algae with biological activities, such as antitumor properties, are greatly reported in the scientific literature. In the present study, a sulfated heterorhamnan obtained from the green seaweed Gayralia brasiliensis (Gb1 fraction) was chemically characterized and its biological activities in the B16-F10 murine melanoma cell line were evaluated. The Gb1 polysaccharidic fraction tested concentrations presented low or absence of cytotoxicity to B16-F10 cells and neither cell proliferation nor cell cycle were altered. Interestingly, Gb1 treatment decreased B16-F10 cells migration and invasion capabilities and CD44 labeling, showing to be a promising compound for further in vitro and in vivo antitumor studies.

Purpose: This study aimed to evaluate a cell therapy strategy with human neural precursor cells (hNPCs) to treat diabetic retinopathy (DR) in Wistar rats induced to diabetes by injecting streptozotocin. Material and methods: Wharton's Jelly Mesenchymal stem cells (WJ-MSCs) were isolated, expanded, and seeded onto a biopolymer substrate to develop neurospheres and obtain the hNPCs. The animals were divided into three groups; non-diabetic (ND) n = four; diabetic without treatment (DM) n = nine; and diabetic with cell therapy (DM + hNPCs) n = nine. After eight weeks of diabetes induction and DR characteristics installed, intravitreal injection of hNPCs (1 x 106 cel/µL) was performed in the DM + hNPCs group. Optical Coherence Tomography (OCT) and Electroretinography (ERG) evaluations were before and during diabetes and after cell therapy. Four weeks post-treatment, histopathological and immunohistochemistry analyses were performed. Results: The repair of the retinal structures in the treated group (DM + hNPCs) was observed by increased thickness of neuroretinal layers, especially in the ganglion cell and photoreceptor layers, higher ERG oscillatory potentials (OPs) amplitudes, and transplanted hNPCs integration into the Retinal Pigment Epithelium. Conclusions: The results indicate that hNPCs reduced DR progression by a neuroprotective effect and promoted retinal repair, making them potential candidates for regenerating the neuroretinal tissue.

Background Considering the expressive number of individuals being diagnosed with diabetes worldwide, it is relevant to find medicines and treatments, in order to achieve diabetes complications, as diabetic retinopathy (DR) long-awaited regression and/or cure. The study aimed to evaluate cell therapy with human neural precursor cells (hNPCs) on induced diabetic retinopathy (DR) in Wistar rats. Methods Wharton's Jelly Mesenchymal stem cells (WJ-MSCs) were isolated, expanded, and seeded onto a biopolymer substrate without growth factors to develop neurospheres, then hNPCs were obtained and characterized by immunocytochemistry. The animals were divided into three groups; non-diabetic (ND) n = 4; diabetic without treatment (DM) n = 9; diabetic with cell therapy (DM + hNPCs) n = 9. Cells were transplanted by intravitreal injection (1 x 106 cel/µL) into each of Streptozotocin (STZ) induced diabetes rats. Evaluations by Optical Coherence Tomography (OCT) and Electroretinography (ERG) were done before and after diabetes induction and post cell therapy. Four weeks after treatment, eye enucleation allowed histopathological and immunohistochemistry (IHC) analysis. Results hNPCs increased the number of retina ganglion cells, ameliorated the photoreceptor layer, and decreased the microvessel points, evidenced by ERG, OCT, histopathological, and IHC findings. The most relevant differences in morphological analysis (treated vs untreated), exhibit the retinal improvement in many layers, notably in the retinal pigment epithelium and photoreceptors. Conclusions hNPCs reduced DR progression, as demonstrated by a neuroprotective effect and promotion of retinal regeneration.

Background In Alzheimer's Disease there is an impairment of the cholinergic system, causing loss of neurons, impairment of intellectual abilities. In this context, the mesenchymal stem cells (MSC) and its applications in cell therapies become target of the research, which may contribute to the treatment of neurodegenerative diseases. Through the differentiation potential of MSCs, prospecting functional repair of the injured tissue from cholinergic neuronal cells could be a potential treatment. The aims was to evaluate the possibility of differentiation of MSCs from the human umbilical cord in nestin-positive neural precursor cells (NPCN+) through the NFBX into cholinergic ‘like’ cells. Methods The isolation of hMSCs from Wharton's jelly (WJ) was by the explant and mononuclear cells by density gradient. hMSCs were plating in natural matrix as NFBX for neurospheres production. Neural precursor cells were subjected to standard cholinergic differentiation protocol. Dissociated neurospheres, neural precursor cells and cholinergic-like cells were characterized by immunocytochemistry. The RT-PCR was done. Results hMSCs were CD73+, CD90+, CD105+, CD34- and CD45- and demonstrated the trilineage differentiation. Neurospheres and their isolated cells were nestin positive, and also expressed NESTIN, MAP2, ßIII-TUBULIN, GFAPgenes. Neural precursor cells that were differentiated in cholinergic-like cells expressed ßIII-TUBULIN protein and choline acetyltransferase enzyme. Conclusions hMSCs on the natural matrix were capable of differentiating hMSC into neurospheres, obtaining neural precursor cells without growth factors or gene transfection before cholinergic differentiation.

Periodontitis is a prevalent disease characterized by the loss of periodontal supporting tissues, bone, periodontal ligament, and cementum. The application of a bone tissue engineering strategy with Decellularized Human Amniotic Membrane (DAM) with adipose-derived stromal cells (ASCs) has shown to be convenient and valuable. This study aims to investigate the treatments of a rat periodontal furcation defect model with DAM, ASCs, and a mineralized extracellular matrix (ECM). Rat ASCs were expanded, cultivated on DAM, and with a bone differentiation medium for four weeks, deposited ECM on DAM. Periodontal healing for four weeks was evaluated by micro-computed tomography and histological analysis after treatments with DAM, ASCs, and ECM and compared to untreated defects on five consecutive horizontal levels, from gingival to apical. The results demonstrate that DAM preserves its structure during cultivation and healing periods, supporting cell attachment, permeation, bone deposition on DAM, and periodontal regeneration. DAM and DAM+ASCs enhance bone healing compared to the control on the gingival level. In conclusion, DAM with ASC or without cells and the ECM ensures bone tissue healing. The membrane supported neovascularization and promoted osteoconduction.

Aim: In vitro evaluation of the influence of bovine xenogenic biomaterials on stem cells from human exfoliated deciduous teeth (SHEDs). The study was divided into three groups: 1) group C (control), containing only MSCs; 2) group BP, containing MSCs and Bonefill Porous®; 3) group BO, containing MSCs and Bio-Oss®. MSCs were derived from a deciduous tooth from a 7-year-old male donor. An aliquot of cells was subjected to immunophenotyping by flow cytometry. Cell viability (neutral red), cytotoxicity (MTT), and cell proliferation (crystal violet) assays were performed. All groups underwent morphological analysis by light microscopy (LM), and the biomaterial with superior performance was submitted to evaluation by scanning electron microscopy (SEM). Time points of 24, 48, and 72 h of culture were used. All results were evaluated with a significance level of 0.05. Results showed that both biomaterials maintained cell viability and cytotoxicity similar to the control. The BO group showed smaller cell proliferation compared to the other groups. In LM evaluation, the BP group showed more spread and adherent cells than the BO group. In SEM, cells of the BP group showed characteristics of more active cells than those of the control. Bovine xenogenic biomaterials positively influenced SHEDs, while the BP group seemed to present higher potential with SHEDs for future application within in vivo and/or clinical studies. Objetivo: Evaluación in vitro de la influencia de los biomateriales xenógenos bovinos en células madre de dientes caducifolios exfoliados (SHEDs) humanos. El estudio se dividió en tres grupos: 1) grupo C (control), que contiene sólo MSCs; 2) grupo BP, que contiene MSCs y Bonefill Porous®; 3) Grupo BO, que contiene MSCs y Bio-Oss®. Los MSC se derivaron de un diente caducifolio de un donante de 7 años. Un conjunto celular fue sometido a inmunofennotización por citometría de flujo. Se realizaron pruebas de viabilidad celular (rojo neutro), citotoxicidad (TM) y proliferación celular (cristal violeta). Todos los grupos fueron sometidos a análisis morfológicos por microscopía ligera (ML), y el biomaterial con rendimiento superior fue sometido a evaluación mediante microscopía electrónica de barrido (SEM). Se utilizaron puntos de tiempo de 24, 48 y 72 horas de cultura. Todos los resultados fueron evaluados con un nivel de significancia de 0,05. Los resultados mostraron que ambos biomateriales mantenían la viabilidad celular y la citotoxicidad similar al control. El grupo bo presentó una menor proliferación celular en comparación con los otros grupos. En la evaluación lm, el grupo BP presentó células más difundidas y adherentes que el grupo bo. En SEM, las células del grupo BP presentaron más características celulares activas que las del control. Los biomateriales xenógenos bovinos influyeron positivamente en los SHEDs, mientras que el grupo BP parecía presentar un mayor potencial con SHEDs para futuras aplicaciones en estudios clínicos y/o in vivo. Objetivo: Avaliação in vitro da influência de biomateriais xenogênicos bovinos sobre células-tronco de dentes decíduos esfoliados humanos (SHEDs). O estudo foi dividido em três grupos: 1) grupo C (controle), contendo apenas CTMs; 2) grupo BP, contendo MSCs e Bonefill Porous®; 3) grupo BO, contendo MSCs e Bio-Oss®. As CTMs foram derivadas de um dente decíduo de um doador de 7 anos de idade. Uma alíquota de células foi submetida à imunofenotipagem por citometria de fluxo. Foram realizados ensaios de viabilidade celular (vermelho neutro), citotoxicidade (MTT) e proliferação celular (cristal violeta). Todos os grupos foram submetidos à análise morfológica por microscopia de luz (ML), e o biomaterial com desempenho superior foi submetido à avaliação por microscopia eletrônica de varredura (MEV). Foram utilizados pontos temporais de 24, 48 e 72 horas de cultura. Todos os resultados foram avaliados com nível de significância de 0,05. Os resultados mostraram que ambos os biomateriais mantiveram a viabilidade celular e citotoxicidade semelhantes ao controle. O grupo BO apresentou proliferação celular menor em comparação aos demais grupos. Na avaliação LM, o grupo BP apresentou mais células disseminadas e aderentes do que o grupo BO. No MEV, as células do grupo BP apresentaram características de células mais ativas do que as do controle. Biomateriais xenogênicos bovinos influenciaram positivamente os SHEDs, enquanto o grupo BP pareceu apresentar maior potencial com SHEDs para futura aplicação em estudos in vivo e / ou clínicos.

A water-soluble sulfated heterorhamnan (Gb1) was isolated from the green seaweed Gayralia brasiliensis and purified by ultrafiltration, yielding a homogeneous polysaccharide (Gb1r). Both fractions contained rhamnose, xylose, galacturonic and glucuronic acids, galactose, and glucose. Chemical and spectroscopic methods allowed the determination of Gb1 and Gb1r chemical structure. Their backbones were constituted by 3-, 2-, and 2,3-linked rhamnosyl units (1:0.49:0.13 and 1:0.58:0.17, respectively), which are unsulfated (13.5 and 14.6%), disulfated (16.6 and 17.8%) or monosulfated at C-2 (8 and 8.6%) and C-4 (24.5 and 23.4%). Gb1 was oversulfated giving rise to Gb1-OS, which presented ~2.5-fold higher content of disulfated rhamnosyl units than Gb1, as determined by methylation analyses and NMR spectroscopy. Gb1 and Gb1-OS potently reduced the viability of U87MG human glioblastoma cells. Gb1 caused cell cycle arrest in the G1 phase, increased annexin V-stained cells, and no DNA fragmentation, while Gb1-OS increased the percentage of cells in the S and G2 phases and the levels of fragmented DNA and cells double-stained with annexin V/propidium iodide, suggesting an apoptosis mechanism. The results suggest that the different effects of Gb1 and Gb1-OS were related to differences in the sulfate content and position of these groups along the polysaccharide chains.

Biological scaffolds have become an attractive approach for repairing the infarcted myocardium and have been shown to facilitate constructive remodeling in injured tissues. This study aimed to investigate the possible utilization of bacterial cellulose (BC) membrane patches containing cocultured cells to limit myocardial postinfarction pathology. Myocardial infarction (MI) was induced by ligating the left anterior descending coronary artery in 45 Wistar rats, and patches with or without cells were attached to the hearts. After one week, the animals underwent echocardiography to assess for ejection fraction and left ventricular end-diastolic and end-systolic volumes. Following patch formation, the cocultured cells retained viability of &gt, 90% over 14 days in culture. The patch was applied to the myocardial surface of the infarcted area after staying 14 days in culture. Interestingly, the BC membrane without cellular treatment showed higher preservation of cardiac dimensions, however, we did not observe improvement in the left ventricular ejection fraction of this group compared to coculture-treated membranes. Our results demonstrated an important role for BC in supporting cells known to produce cardioprotective soluble factors and may thus provide effective future therapeutic outcomes for patients suffering from ischemic heart disease.

International audience; p-Cresyl sulfate (PCS), indoxyl sulfate (IS), and inorganic phosphate (Pi) are uremic toxins found in chronic kidney disease (CKD) that are closely related to endothelial extracellular vesicles (EVs) formation. The present study aimed to understand the role of EVs and their role in cell adhesion and migration, inflammation, and oxidative stress. Human endothelial cells were treated with PCS, IS, and Pi in pre-established uremic and kinetic recommendations. EVs were characterized using scanning electron microscopy, flow cytometry, and NanoSight assays. The concentrations of EVs were established using Alamar Blue and MTT assays. Cell adhesion to extracellular matrix proteins was analyzed using an adhesion assay. Inflammation and oxidative stress were assessed by vascular cell adhesion molecule-1 expression/monocyte migration and reactive oxygen species production, respectively. The capacity of EVs to stimulate endothelial cell migration was evaluated using a wound-healing assay. Our data showed that endothelial cells stimulated with uremic toxins can induce the formation of EVs of different sizes, quantities, and concentrations, depending on the uremic toxin used. Cell adhesion was significantly (P < 0.01) stimulated in cells exposed to PCS-induced extracellular vesicles (PCSEVs) and inorganic phosphate-induced extracellular vesicles (PiEVs). Cell migration was significantly (P < 0.05) stimulated by PCSEVs. VCAM-1 expression was evident in cells treated with PCSEVs and IS-induced extracellular vesicles (ISEVs). EVs are not able to stimulate monocyte migration or oxidative stress. In conclusion, EVs may be a biomarker of endothelial injury and the inflammatory process, playing an important role in cell-to-cell communication and pathophysiological processes, although more studies are needed to better understand the mechanisms of EVs in uremia.

Uremic toxins, such as p-cresyl sulfate (PCS) and indoxyl sulfate (IS), contribute to endothelial dysfunction in chronic kidney disease (CKD). This process is mediated by several cellular pathways, but it is unclear whether cAMP-responsive element-binding protein (CREB) and activating transcription factor 1 (ATF1) participate in endothelial dysfunction in uremic conditions despite playing roles in inflammatory modulation. This study aimed to evaluate the expression, activation, and transcriptional activity of CREB/ATF1 in endothelial cells exposed to PCS, IS, and uremic serum (US). In vitro, ATF1 protein levels were increased by PCS and IS, whereas CREB levels were enhanced only by IS. Activation through CREB-Ser

Lipopolysaccharide (LPS) induces fever through cytokines like receptor-activator of nuclear factor kappa B ligand (RANKL), triggering mediators like prostaglandins (PG), endothelin-1 (ET-1), corticotrophin-releasing factor (CRF), substance P (SP) and endogenous opioids. LPS-induced fever is reduced in females compared with males except in ovariectomized (OVX) females which show increased fever mediated by PG. The present study aimed to identify the mediators involved in fever in intact and OVX female rats. Fever was induced with LPS (50 mu g/kg) intraperitoneally or CRF (2.5 mu g), ET-1 (1 pg), morphine (10 mu g) and SP (500 ng) intracerebroventricularly in sham-operated and OVX rats. The role of RANKL was evaluated with osteoprotegerin (OPG, 1 mu g, intracerebroventricularly). Expression of RANK, CRFI/II, ETB, mu-opioid (MOR) and NK1 receptors was evaluated by confocal microscopy. Besides LPS, only morphine induced fever in OVX rats while all mediators induced fever in sham-operated animals. OPG abolished LPS-induced fever in OVX but not sham-operated animals. Overall, fever involves similar central mediators in cycling females and males but only morphine induced fever in OVX females. Importantly, RANK/RANKL participates in LPS-induced fever in OVX females, as in males but not in cycling females. Funding Agencies|Fundacao de Amparo a Pesquisa do Maranhao (FAPEMA/UNIVERSAL) [00431/12]; Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES/CAPES Sandwich Program) [007414/2014-05]; Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)National Council for Scientific and Technological Development (CNPq)

An experiment was designed to determine the influence of fibre and betaine on the development of the intestine, liver and pancreas of broilers from hatch to 14 d of age. A total of 250-day-old Cobb 500 male broilers were allocated to 16 cages with 15 broilers each. Treatments were arranged in a 2 × 4 factorial design, consisting of 2 feed formulations (low and high fibre diets) and 4 levels of betaine (0, 1, 3 or 5 kg/t). At hatch, 10 birds in total were euthanised, and samples of the liver, pancreas, yolk sac and intestine were collected for reference of the analysed parameters before the start of the trial. On d 4, 9 and 14, 5 birds per cage (10 birds per treatment) were selected, euthanised and treated as the same as the birds at hatch. Villus height and width and crypt depth were determined on the duodenum samples, and absorptive area was calculated. The number of enterocytes in mitosis at the villus was determined by a positive reaction to antibody for Ki67 protein, and fused villus was evaluated visually. The relative weight of the yolk sac reduced (P

A low-molecular-weight (LMW) heterofucan (designated fucan B) was obtained from the brown seaweed, Spatoglossum schroederi, and its activity as an inhibitor of capillary-like tube formation by endothelial cells (ECs) was analyzed. Chemical, infrared and electrophoretic analyses confirmed the identity of fucan B. In contrast to other LMW fucans, fucan B (0.012–0.1 mg/mL) inhibited ECs capillary-like tube formation in a concentration-dependent manner. In addition, fucan B (0.01–0.05 mg/mL) did not affect ECs proliferation. Fucan B also inhibited ECs migration on a fibronectin-coated surface, but not on laminin- or collagen-coated surfaces. Biotinylated fucan B was used as a probe to identify its localization. Confocal microscopy experiments revealed that biotinylated fucan did not bind to the cell surface, but rather only to fibronectin. Our findings suggest that fucan B inhibits ECs capillary-like tube formation and migration by binding directly to fibronectin and blocking fibronectin sites recognized by cell surface ligands. However, further studies are needed to evaluate the in vivo effects of fucan B.

Piper caldense C. DC., Piperaceae, commonly known as “pimenta-d’água”, “pimenta-darda” or “paguarandy” in Brazil, is a shrub that grows mainly in humid and shaded habitats. The present study investigates the anatomy of the leaves and stems of P. caldense by light and scanning electron microscopy in order to provide supporting data for correct identification of the species. The leaves are hypostomatic, have a 2-layered hypodermis, and posses pearl glands. The midrib shows a ‘U’-shaped stele comprised of about ten collateral vascular bundles. The main anatomical marker of the stem is the presence of a continuous sclerenchymatous sheath in the pith. Two forms of calcium oxalate crystals, namely crystal sand and raphides, are observed in this species. Keywords: Anatomy, Pimenta-d’água, Piperaceae, Light and scanning electron microscopy

The scaffolds and their interaction with mesenchymal stem cells are objects of study in bioengineering and tissue repair. Mechanisms such as surface adhesion, proliferation, viability, and cytotoxicity are essential for the development of therapies. The present study analyzed the influence of platelet-rich fibrin (PRF) in viability, cytotoxicity, and proliferation of stem cells from human exfoliated deciduous teeth (SHED) exposed to bovine biomaterial surfaces. The studied groups were divided and analyzed as follows: (S) only SHED as control Group; (SB) SHED + biomaterial; (SBP) SHED + biomaterial + PRF. Analyses of cells seeded in 24-well plates were performed after 24, 48 and 72 hours. Individual groups were subjected to viability, cytotoxicity and cell proliferation tests using neutral red, MTT and crystal violet, respectively; and in the 72-hour group, scanning electron microscopy (SEM) was performed to record cell ultra-morphology. Data were submitted to statistical analysis by two-factor ANOVA with a significance level of 5%. The results demonstrated a better performance in the viability/cytotoxicity and proliferation of stem cells for the group (SBP) in comparison to the group (SB) and the group (S). The applied statistical tests showed that the biomaterial factor, time, and interaction between them gave rise to results with statistical significance. SHED submitted to bovine biomaterial were more viable, proliferative and with lower toxicity when associated with PRF. PRF seemed to activate the metabolism of stem cells in culture, indicating that such an association can bring an effective benefit in clinical outcome. El marco y su interacción con las células madres mesenquimales son objetos de estudio en bioingeniería y reparación de tejidos. Mecanismos como la adhesión superficial, la proliferación, la viabilidad y la citotoxicidad son fundamentales para el desarrollo de terapias. El presente estudio analizó la influencia de la fibrina rica en plaquetas (PRF) sobre la viabilidad, citotoxicidad y proliferación de células madre mesenquimales de dientes primarios humanos exfoliados (SHED) en contacto con biomateriales bovinos. Los grupos estudiados se dividieron y analizaron de la siguiente manera: (S) solo SHED como grupo de control; (SB) SHED + biomaterial; (SBP) SHED + biomaterial + PRF. Se realizaron análisis de células sembradas en placas de 24 pocillos después de 24, 48 y 72 horas. Los grupos individuales se sometieron a pruebas de viabilidad, citotoxicidad y proliferación celular utilizando rojo neutro, MTT y violeta cristal, respectivamente, y en el grupo de 72 horas, se realizó microscopía electrónica de barrido (SEM) para registrar la morfología celular. Los datos se sometieron a análisis estadístico mediante ANOVA de dos factores con un nivel de significancia del 5%. Los resultados demostraron un mejor desempeño en la viabilidad / citotoxicidad y proliferación de células madre para el grupo (SBP) en comparación con el grupo (SB) y el grupo (S). Las pruebas estadísticas aplicadas mostraron que el factor biomaterial, el tiempo y la interacción entre ellos dieron lugar a resultados con significación estadística, las SHED sometidas a biomaterial bovino fueron más viables, proliferativos y con menor toxicidad cuando se asociaron con PRF. PRF parecio activar el metabolismo de las células madres en cultivo, lo que indica que tal asociación puede aportar un beneficio efectivo en los resultados clínicos. O arcabouço e a sua interação com as células-tronco mesenquimais são objetos de estudo em bioengenharia e reparo de tecidos. Mecanismos como adesão superficial, proliferação, viabilidade e citotoxicidade são fundamentais para o desenvolvimento de terapias. O presente estudo analisou a influência da fibrina rica em plaquetas (PRF) na viabilidade, citotoxicidade e proliferação de células-tronco mesenquimais de dentes decíduos humanos esfoliados (SHED) em contato com biomaterial de origem bovina. Os grupos estudados foram divididos e analisados ​​da seguinte forma: (S) apenas SHED como Grupo controle; (SB) SHED + biomaterial; (SBP) SHED + biomaterial + PRF. As análises das células semeadas em placas de 24 poços foram realizadas após 24, 48 e 72 horas. Grupos individuais foram submetidos a testes de viabilidade, citotoxicidade e proliferação celular usando vermelho neutro, MTT e cristal violeta, respectivamente, e no grupo de 72 horas foi realizada a microscopia eletrônica de varredura (MEV) para registrar a ultra morfologia celular. Os dados foram submetidos à análise estatística por ANOVA de dois fatores com nível de significância de 5%. Os resultados demonstraram um melhor desempenho na viabilidade / citotoxicidade e proliferação de células-tronco para o grupo (SBP) em relação ao grupo (SB) e ao grupo (S). Os testes estatísticos aplicados mostraram que o fator biomaterial, o tempo e a interação entre eles deram origem a resultados com significância estatística. As SHED submetidas ao biomaterial bovino se apresentaram mais viáveis, proliferativas e com menor toxicidade quando associadas à PRF. A PRF pareceu ativar o metabolismo das células-tronco em cultivo, indicando que tal associação pode trazer um benefício efetivo nos desfechos clínicos.

Made available in DSpace on 2022-05-01T08:44:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-08-01 This study aimed to differentiate human mesenchymal stem cells (hMSCs) from the human umbilical cord in cholinergic-like neurons using a natural membrane. The isolation of hMSCs from Wharton’s jelly (WJ) was carried out using “explant” and mononuclear cells by the density gradient from umbilical blood and characterized by flow cytometry. hMSCs were seeded in a natural functional biopolymer membrane to produce neurospheres. RT-PCR was performed on hMSCs and neurospheres derived from the umbilical cord. Neural precursor cells were subjected to a standard cholinergic-like neuron differentiation protocol. Dissociated neurospheres, neural precursor cells, and cholinergic-like neurons were characterized by immunocytochemistry. hMSCs were CD73+, CD90+, CD105+, CD34-and CD45-and demonstrated the trilineage differentiation. Neurospheres and their isolated cells were nestin-positive and expressed NESTIN, MAP2, ßIII-TUBULIN, GFAP genes. Neural precursor cells that were differentiated in cholinergic-like neurons expressed ßIII-TUBULIN protein and choline acetyltransferase enzyme. hMSCs seeded on the natural membrane can differentiate into neurospheres, obtaining neural precursor cells without growth factors or gene transfection before cholinergic phenotype differentiation. Advanced Therapy and Cellular Biotechnology in Regenerative Medicine Department Child and Adolescent Health Research and Pequeno Príncipe Faculties Pelé Pequeno Príncipe Institute Cell Biology Department Federal University of Paraná Research Center in Applied Morphology and Immunology NuPMIA Faculty of Medicine University of Brasília Biodiversity and Biotechnology Research Parnaíba Delta Federal University Faculty of Pharmaceutics Sciences São Paulo State University (UNESP) Physics Department São Paulo State University (UNESP) Laboratory Biosurgical Research Cardiovascular Division Pompidou Hospital University of Paris Faculty of Pharmaceutics Sciences São Paulo State University (UNESP) Physics Department São Paulo State University (UNESP)

A bichectomia é um procedimento que se constitui na remoção da porção anterior do corpo adiposo de Bichat (paCAB). O extrato celular obtido após o processamento por meio de digestão enzimática desse tecido é chamado de fração vascular estromal (FVE). A natureza das células que compõem a FVE qualifica esse produto para uma ampla gama de aplicações clínicas, especialmente em procedimentos de estímulo à renovação e à reparação tecidual, incluindo-se os fins estéticos de rejuvenescimento em face. Entretanto, o aproveitamento desse material biológico vivo está diretamente relacionado ao seu adequado manejo e transporte, desde sua coleta, processamento e envio para aplicação clínica. Este trabalho teve como objetivo caracterizar a FVE obtida a partir da paCAB, e também verificar se o plasma pobre em plaqueta (PPP) autólogo é eficiente na manutenção da viabilidade celular, podendo ser uma opção de meio para transporte até sua aplicação clínica mediata. Três pacientes com indicação da realização de bichectomia participaram da pesquisa. Antes da remoção da paCAB foi realizada a coleta sanguínea venosa para a obtenção do PPP. A paCAB bilateral coletada foi enviada para o Centro de Processamento Celular Curityba Biotech e submetida ao protocolo para desagregar as células da matriz extracelular. Ao final, a FVE foi fracionada em alíquotas que foram acondicionadas em seringas e em placas de cultivo, e impostas a ensaios para se avaliar a viabilidade celular nos tempos 0, 24 e 48 h. A viabilidade celular e a caracterização das células presentes na FVE foram avaliadas por microscopia de luz e imunofenotipagem por citometria de fluxo. Uma amostra foi mantida em garrafa de cultivo até atingir 7x106 células. Após, foram impostas a comprovação da presença de células-tronco mesenquimais (CTM) capazes de se manter em cultivo padrão. A amostra analisada por imunofenotipagem confirmou a existência das seguintes células: células-tronco mesenquimais e hematopoiéticas, células endoteliais e células T.

Hemoplasmas were detected in two apparently healthy captive South American coatis (Nasua nasua) from southern Brazil during an investigation for vector-borne pathogens. Blood was subjected to packed cell volume (PCV) determination, a commercial real-time PCR panel for the detection of Anaplasma spp., Babesia spp., Bartonella spp., Hepatozoon spp., Leishmania spp., Mycoplasma haemofelis, 'Candidatus Mycoplasma turicensis', 'Candidatus Mycoplasma haemominutum', Neorickettsia risticii, Rickettsia rickettsii and Leptospira spp., and a pan-hemoplasma conventional PCR assay. PCV was normal, but both coatis tested positive for hemoplasmas and negative for all the remaining pathogens tested. Using different techniques for microscopy (light, confocal or SEM), structures compatible with hemoplasmas were identified. Sequencing of the 16S rRNA gene identified an organism resembling Mycoplasma haemofelis and another hemotropic Mycoplasma sp., with a sequence identity of 96.8% to a Mycoplasma sp. previously detected in capybaras.

Piper solmsianum C.DC., which is popularly known as pariparoba, is a shrub that measures 1-3 m in height and it inhabits areas with wet tropical soils. The objective of this study was to analyze the leaf and stem anatomy using light microscopy, scanning electron micrographs, and energy-dispersive X-ray spectroscopy in order to provide information for species identification. The anatomical profile showed the following main microscopic markers: hypostomatic leaf; hypodermis layer on both sides; pearl glands; biconvex midrib shape; five collateral vascular bundles in open arc with the central bundle larger than the others; circular stem shape; collateral vascular bundles arranged in two rings; sinuous sclerenchymatic sheath in the pith; secretory idioblasts; and starch grains in the mesophyll, in the ground parenchyma of the midrib, petiole, and in the stem; and six morphotypes of calcium oxalate crystals (styloids, cuneiform, tabular crystal rosettes, cuneiform crystal rosettes, elongated square dipyramids, as well as very elongated square dipyramids).

Traditional uses, common names, adulterations and substitutions of medicinal plants are some of the considerations that require qualitative analysis for effective quality control. Microscopy studies of selected “boldo” species using light and scanning electron microscopy reveled key diagnostic traits that can help species identification. These traits can contribute to the quality control of the botanical traded in the form of whole, fragmented or powdered leaves. The studied species exhibited different features in their leaf trichome micro-morphology; Peumus boldus has stellate non-glandular trichomes, Plectranthus neochilus possesses simple non-glandular trichomes with warty cuticle, and short-stalked capitate and peltate glandular trichomes, and Plectranthus barbatus has simple non-glandular with smooth cuticle, short- and long-stalked capitate, as well as peltate glandular trichomes. These differences prove to be helpful in the species identification and quality control of products containing “boldo” species.

Piper amalago L. is a medicinal plant traditionally used as a healing agent for wounds, burns, abscesses, boils, and insect bites. The current study aimed to evaluate the possible effects of the aqueous crude extract obtained from P. amalago leaves, in different concentrations and in different incubation times, using the in vitro model of mouse fibroblasts (3T3). The extract was tested in different concentrations at the 24 h incubation time for analysis of cell viability, cytotoxicity, proliferation, cell morphology, immunostaining, adhesion and cell spreading assays, as well as to determine the hydroxyproline concentration and activity of the metalloproteinase MMP2. Morphologically, after exposure to the concentrations of 15 and 150 µg/mL, the cells maintained the morphology, yet a greater number of cells with more expansions of the cell body and larger than the control cells were observed. The treated cell culture also showed a greater number of cells, larger cells, a greater expansion of the cell body, adherent cells spread over the substrate, and a more juxtaposed, central and spherical nucleus. The treatment induced greater cell adhesion to the polymer, fibronectin, and collagen I. Biochemical results showed a significant increase in the hydroxyproline amino acid after exposure for 96 h. The extract did not induce loss of cell viability until the concentration reached 150 µg/mL, positively modulating proliferation, morphology, adhesion, degree of spreading, and organization of microfilaments. The extract also promoted a significant increase in the hydroxyproline amino acid. Piper amalago L. es una planta medicinal utilizada tradicionalmente como agente cicatrizante de heridas, quemaduras, abscesos, furúnculos y picaduras de insectos. El presente estudio tuvo como objetivo evaluar los posibles efectos del extracto crudo acuoso obtenido de las hojas de P. amalago, en diferentes concentraciones y en diferentes tiempos de incubación, utilizando el modelo in vitro de fibroblastos de ratón (3T3). El extracto se probó a diferentes concentraciones en el tiempo de incubación de 24 horas para el análisis de viabilidad celular, citotoxicidad, proliferación, morfología celular, inmunotinción, ensayos de adhesión y propagación celular, así como para determinar la concentración de hidroxiprolina y la actividad metaloproteinasa de MMP2. Morfológicamente, tras la exposición a concentraciones de 15 y 150 µg/mL, las células mantuvieron su morfología, pero se observó un mayor número de células con más expansiones del cuerpo celular y más grandes que las células control. El cultivo celular tratado también mostró un mayor número de células, células más grandes, mayor expansión del cuerpo celular, células adherentes difundidas sobre el sustrato y núcleos más yuxtapuestos, centrales y esféricos. El tratamiento indujo una mayor adhesión celular al polímero, fibronectina y colágeno I. Los resultados bioquímicos mostraron un aumento significativo en el aminoácido hidroxiprolina después de 96 h de exposición. El extracto no indujo pérdida de viabilidad celular hasta que la concentración alcanzó 150 µg/mL, modulando positivamente la proliferación, morfología, adhesión, grado de diseminación y organización de los microfilamentos. El extracto también promovió un aumento significativo del aminoácido hidroxiprolina. Piper amalago L. é uma planta medicinal tradicionalmente usada como agente de cura para feridas, queimaduras, abcessos, furúnculos e picadas de insetos. O presente estudo teve como objetivo avaliar os possíveis efeitos do extrato bruto aquoso obtido das folhas de P. amalago, em diferentes concentrações e em diferentes tempos de incubação, utilizando o modelo in vitro de fibroblastos de camundongo (3T3). O extrato foi testado em diferentes concentrações no tempo de incubação de 24 horas para análise de viabilidade celular, citotoxicidade, proliferação, morfologia celular, imunomarcação, adesão e ensaios de propagação celular, bem como para determinar a concentração de hidroxiprolina e atividade da metaloproteinase MMP2. Morfologicamente, após exposição às concentrações de 15 e 150 µg/mL, as células mantiveram a morfologia, porém foi observado um número maior de células com mais expansões do corpo celular e maiores que as células controle. A cultura celular tratada também apresentou maior número de células, células maiores, maior expansão do corpo celular, células aderentes espalhadas pelo substrato e núcleo mais justaposto, central e esférico. O tratamento induziu maior adesão celular ao polímero, fibronectina e colágeno I. Os resultados bioquímicos mostraram um aumento significativo no aminoácido hidroxiprolina após exposição por 96 h. O extrato não induziu perda de viabilidade celular até a concentração atingir 150 µg/mL, modulando positivamente a proliferação, morfologia, adesão, grau de disseminação e organização dos microfilamentos. O extrato também promoveu um aumento significativo no aminoácido hidroxiprolina.

The citrus industry is among the most important worldwide, but citrus plants are affected by a large number of diseases, such as Citrus Black Spot (CBS), caused by the fungus Phyllosticta citricarpa. To identify alternative methods for CBS control, endophytic fungi were isolated by our group from healthy citrus plants in Brazil. Over 400 fungal isolates were obtained, and isolate LGMF1254 was selected based on its inhibitory effect on the growth of P. citricarpa because of the production of volatile organic compounds (VOCs). This isolate was identified as Muscodor sp. by morphological examination and phylogenetic analysis using the internal transcribed spacer (ITS) region: High similarity with M. sutura, M. vitigenus, and M. equiseti was observed. We also sequenced the RPB2 gene, and isolate LGMF1254 showed 99% similarity with M. sutura. To identify this isolate at the species level, the main VOCs were determined, and according to this analysis, LGMF1254 may be classified as an M. sutura strain; however, sequences from regions other than ITS and RPB2 are necessary for conclusive genotyping. The VOCs produced by strain LGMF1254 can be considered an alternative way to control P. citricarpa during the transport of fruits and to prevent the development of CBS signs.

RESUMO Tanacetum vulgare L., conhecida popularmente como catinga-de-mulata, é utilizada na medicina tradicional como vermífugo, digestivo e emenagogo. Objetivou-se analisar a morfoanatomia de folhas e caules dessa espécie medicinal com vistas ao controle da qualidade. Foram utilizadas técnicas usuais de microscopia de luz e eletrônica de varredura. As folhas são compostas, pinatissectas e anfiestomáticas. Tricomas tectores flageliformes simples e glandulares capitados são observados. A nervura central tem formato biconvexo, o pecíolo é côncavo-convexo, a ráque é plano-convexa e o caule é arredondado. Feixes vasculares colaterais, calotas de fibras perivasculares e colênquima lamelar estão presentes na folha e no caule. Os caracteres morfoanatômicos evidenciados contribuem na identificação do táxon e fornecem subsídios farmacobotânicos para o controle da qualidade de drogas vegetais e fitoterápicos. ABSTRACT Tanacetum vulgare L., popularly known as tansy, is used in traditional medicine as a vermifuge, digestive and emmenagogue. This study aimed to analyze the morphoanatomical leaves and stems of this medicinal plant in order to control the quality. Usual techniques of light and scanning electron microscopy were used. The leaves are composed, pinatissect and amphistomatic. Simple and flagelliform non- glandular trichomes and capitate glandular trichomes are observed. The midrib has biconvex shape, the petiole is concavo-convex, the rachis is plano-convex, and the stem is rounded. Collateral vascular bundles, perivascular fiber caps and laminar collenchyma are encountered in the leaves and stems. Evidenced the morphological and anatomical features contribute to the identification of the taxon and provide pharmacobotanical data for the quality control of herbal drugs.

For the safe use of nanoparticles, especially in the biomedical field, their stability in different environments and the prevention of binding to the component organisms, which could lead to nanoparticle aggregation, is indispensable. Herein, we present a simple, efficient and biologically based method to obtain small gum arabic (GA)-stabilized gold nanoparticles (GA-AuNPs) with remarkable stability in physiological pHs. The in vitro stability tests in intestinal (pH 6.8) and gastric (pH 1.2) simulated pHs revealed that GA-AuNPs exhibit a surprisingly high stability even near the zero zeta potential. When subjected to GA-AuNPs, changes in the viability, proliferation and morphology were selectively induced in the B16-F10 melanoma cell line, whereas no alterations in the macrophage cell line, RAW 264.7, or in the fibroblast cell line, BALB/3T3, were observed at the same concentrations. Therefore, considering the remarkable stability and selective effect on cell lines, we show that GA-AuNPs exhibit properties that could provide a future alternative for melanoma treatment.

Silver nanoparticles (AgNPs) are widely used in consumer products due to their antibacterial property; however, their potential toxicity and release into the environment raises concern. Based on the limited understanding of AgNPs aggregation behavior, this study aimed to investigate the toxicity of uncoated (uc-AgNP) and coated with polyvinylpyrrolidone (PVP-AgNP), at low concentrations (0.5–100 ng/mL), under dark and visible-light exposure, using a plant test system. We exposed Allium cepa seeds to both types of AgNPs for 4–5 days to evaluate several toxicity endpoints. AgNPs did not cause acute toxicity (i.e., inhibition of seed germination and root development), but caused genotoxicity and biochemical alterations in oxidative stress parameters (lipid peroxidation) and activities of antioxidant enzymes (superoxide dismutase and catalase) in light and dark conditions. However, the light exposure decreased the rate of chromosomal aberration and micronuclei up to 5.60x in uc-AgNP and 2.01x in PVP-AgNP, and 2.69x in uc-AgNP and 3.70x in PVP-AgNP, respectively. Thus, light exposure reduced the overall genotoxicity of these AgNPs. In addition, mitotic index alterations and morphoanatomical changes in meristematic cells were observed only in the dark condition at the highest concentrations, demonstrating that light also reduces AgNPs cytotoxicity. The light-dependent aggregation of AgNPs may have reduced toxicity by reducing the uptake of these NPs by the cells. Our findings demonstrate that AgNPs can be genotoxic, cytotoxic and induce morphoanatomical and biochemical changes in A. cepa roots even at low concentrations, and that visible-light alters their aggregation state, and decreases their toxicity. We suggest that visible light can be an alternative treatment to remediate AgNP residues, minimizing their toxicity and environmental risks.

Endothelial dysfunction in uremia can result in cell-to-cell junction loss and increased permeability, contributing to cardiovascular diseases (CVD) development. This study evaluated the impact of the uremic milieu on endothelial morphology and cell junction&rsquo, s proteins. We evaluated (i) serum levels of inflammatory biomarkers in a cohort of chronic kidney disease (CKD) patients and the expression of VE-cadherin and Zonula Occludens-1 (ZO-1) junction proteins on endothelial cells (ECs) of arteries removed from CKD patients during renal transplant, (ii) ECs morphology in vitro under different uremic conditions, and (iii) the impact of uremic toxins p-cresyl sulfate (PCS), indoxyl sulfate (IS), and inorganic phosphate (Pi) as well as of total uremic serum on VE-cadherin and ZO-1 gene and protein expression in cultured ECs. We found that the uremic arteries had lost their intact and continuous endothelial morphology, with a reduction in VE-cadherin and ZO-1 expression. In cultured ECs, both VE-cadherin and ZO-1 protein expression decreased, mainly after exposure to Pi and uremic serum groups. VE-cadherin mRNA expression was reduced while ZO-1 was increased after exposure to PCS, IS, Pi, and uremic serum. Our findings show that uremia alters cell-to-cell junctions leading to an increased endothelial damage. This gives a new perspective regarding the pathophysiological role of uremia in intercellular junctions and opens new avenues to improve cardiovascular outcomes in CKD patients.

Background Posttransplant cell tracking, via stem cell labeling, is a crucial strategy for monitoring and maximizing benefits of cell-based therapies. The structures and functionalities of polysaccharides, proteins, and lipids allow their utilization in nanotechnology systems. Materials and methods In the present study, we analyzed the potential benefit of curcumin-loaded nanoparticles (NPC) using Vero cells (in vitro) and NPC-labeled adipose-derived mesenchymal stem cells (NPC-ADMSCs) (in vivo) in myocardial infarction and sciatic nerve crush preclinical models. Thereafter, transplantation, histological examination, real time imaging, and assessment of tissue regeneration were done. Results Transplanted NPC-ADMSCs were clearly identified and revealed potential benefit when used in cell tracking. Conclusion This approach may have broad applications in modeling labeled transplanted cells and in developing improved stem cell therapeutic strategies., Video abstract

In southern Brazil, the bottled latex of Synadenium grantii Hook f., Euphorbiaceae, is popularly used as a treatment of all types of cancer. Similarly, Synadenium umbellatum Pax. is used in the central western region of Brazil for the same purpose and in the same manner of use. Both plants are popularly known as janaúba or leitosinha. The objectives of this study were to use pharmacobotanical analysis to verify whether these two species, which are considered to be distinct, are actually the same to determine anatomical markers; to assist in the identification and differentiation of other Euphorbia; and to evaluate the cytotoxic activity of the latex in relation to HeLa and HRT-18 cells. Leaves and stems of the species were collected in Goiânia and Ponta Grossa and were investigated using scanning electron microscopy and optical microscopy techniques. The latex was also collected and analyzed in relation to its cytotoxic effect by employing MTT and NR techniques. The pharmacobotanical study of the specimens in both localities showed that they were the same species, namely Euphorbia umbellata (Pax) Bruyns, which is the scientific nomenclature accepted and confirmed by an expert taxonomist who specializes in Euphorbia. The pharmacobotanical characteristics highlighted in this study can assist in the identification of the taxon and contribute to the control of the quality of this plant drug. The evaluation of the latex in relation to HRT-18 cells demonstrated action after 48 h of experiment. In contrast, in relation to HeLa cells its induced cytotoxicity in all times and a dose-dependent manner. The IC50 values (72 h) observed were 252.58 ± 18.51 μg/ml and 263.42 ± 15.92 μg/ml to MTT experiment and 250.18 ± 19.48 μg/ml and 430.56 ± 19.71 μg/ml to NR experiment for the HeLa and HRT-18 cells, respectively. Keywords: HeLa, HRT-18, Morfo-anatomy, Synadenium grantii, Synadenium umbellatum

Piper amalago L., Piperaceae, popularly known as jaborandi-manso, is a shrub that spans a height of 2–7 m. It can be found in the regions of Southern America downward up to the south of Brazil. Traditionally it is used to treat digestive problems, heart problems, and burns. This study aims to conduct an anatomical investigation and analysis of the leaves and stems of P. amalago through electron scanning and optical micro techniques. The analysis showed that P. amalago has a hypostomatic leaf, with a subepidermal layer on its surface. There are grandular trichomes that resemble sacs, conic non-glandular trichomes, dorsiventral mesophyll, and a plano-convex midrib having a single vascular bundle in the center. The petiole is short with irregularly shaped and adaxially grooved. The stem is circular in shape and contains two circles of vascular bundles and a sclerenchymatic sheath in the perimedular region. These anatomical features of the Piper amalago's leaves and stems make it easy to pick it out among other species of the Piper genus. This is helpful when conducting quality control process. Keywords: Anatomy, Quality control, Jaborandi-manso, Piper amalago, Piperaceae

This study evaluated the physicochemical characterization and rheological behavior of gabiroba pulp, and a gabiroba jam formulation. Gabiroba pulp presented a heterogeneous ultrastructure with a denser area formed by a compact mesh and a porous interface containing fibers. The fibers’ presence promoted a slip effect when the gabiroba pulp was subjected to shear. Gabiroba pulp showed a gel behavior with thermal stability. Gabiroba jam, developed using pulp as the raw material, had shear thinning behavior exhibiting yield stress described by the Herschel-Bulkley model. The dynamic oscillatory analysis showed that gabiroba jam typically behaved like a gel, i.e., G′ values higher than the G″ in all frequency ranges evaluated. The results showed that gabiroba pulp is suitable for use as a raw material in the development of food products such as jam, encouraging the preservation of this native Brazilian species.

A heteropolysaccharide was isolated by cold aqueous extraction from edible mushroom Pleurotus eryngii ("King Oyster") basidiocarps and its biological properties were evaluated. Structural assignments were carried out using mono- and bidimensional NMR spectroscopy, monosaccharide composition, and methylation analyses. A mannogalactan having a main chain of (1→6)-linked α-d-galactopyranosyl and 3-O-methyl-α-d-galactopyranosyl residues, both partially substituted at OH-2 by β-d-Manp (MG-Pe) single-unit was found. Biological effects of mannogalactan from P. eryngii (MG-Pe) were tested against murine melanoma cells. MG-Pe was non-cytotoxic, but reduced in vitro melanoma cells invasion. Also, 50mg/kg MG-Pe administration to melanoma-bearing C57BL/6 mice up to 10days decreased in 60% the tumor volume compared to control. Additionally, no changes were observed when biochemical profile, complete blood cells count (CBC), organs, and body weight were analyzed. Mg-Pe was shown to be a promising anti-melanoma molecule capable of switching melanoma cells to a non-invasive phenotype with no toxicity to melanoma-bearing mice.

Fucan is a term used to denominate sulfated l-fucose rich polysaccharides. Here, a heterofucan, named fucan B, was extracted from the Spatoglossum schröederi seaweed. This 21.5kDa galactofucan inhibited CHO-K1 proliferation and migration when fibronectin was the substrate. Fucan B derivatives revealed that such effects depend on their degree of sulfation. Fucan B did not induce cell death, but promoted G1 cell cycle arrest. Western blotting and flow cytometry analysis suggest that fucan B binds to fibronectin and activates integrin, mainly integrin α5β1, which induces FAK/RAS/MEK/ERK activation. FAK activation inhibits CHO-K1 migration on fibronectin and ERK blocks cell cycle progression. This study indicates that fucan B could be applied in developing new antitumor drugs.

Baccharis rufescens Spreng. var. tenuifolia (DC.) Baker pertence à família Asteraceae e é usada na medicina tradicional como estomáquico e hepatoprotetor. Um estudo fitoquímico mostrou a presença de flavonoides e triterpenos, sendo que os extratos clorofórmico e metanólico de folhas apresentaram-se ativos no bioensaio de toxicidade sobre Artemia salina Leach. Além disso, extratos clorofórmicos evidenciaram a presença de peróxidos, sugerindo sua aplicação no tratamento da malária (Schenkel at al., 2002; Montanher et al., 2002; Moreira et al., 2003). Considerando a importância farmacológica de B. rufescens var. tenuifolia, o presente trabalho objetivou o estudo morfoanatômico e histoquímico do caule e da folha dessa espécie, a fim de fornecer subsídios farmacognósticos para o controle de qualidade. O material botânico foi submetido às técnicas usuais empregadas na microscopia de luz e microscopia eletrônica de varredura. Folha anfiestomática, presença de estômatos anomocíticos, tricomas glandulares capitados bisseriados, tricomas tectores flageliformes simples unisseriados, dutos secretores associados ao floema, calota de fibras perivasculares e cristais de oxalato de cálcio do tipo estiloide e prismático na região medular do caule foram as principais características observadas que auxiliam na identificação do táxon.

There are differences in the immune response, and particularly fever, in males and females. Here we investigated how the febrile response induced by lipopolysaccharide (LPS) and different endogenous pyrogens were affected by female gonadal hormones. The febrile response to intraperitoneal injection of LPS (50 μg/kg) was 40% lower in female rats when compared with male or ovariectomized (OVX) female rats. Accordingly, oestrogen replacement in OVX animals reduced LPS-induced fever. The treatment with the prostaglandin (PG) synthesis inhibitor indomethacin (2 mg/kg, i.p. 30 min before) reduced the febrile response induced by LPS in both, OVX (88%) and sham-operated (71%) rats. In line with the enhanced fever in OVX rats, they showed an increased expression of cyclooxygenase-2 (COX-2) in the hypothalamus and elevated levels of prostaglandin E2 (PGE2). In addition, OVX rats were hyper-responsive to PGE2 injected intracerebroventricularly (i.c.v.). In strong contrast to the enhanced fever in response to LPS and PGE2, the febrile response induced by i.c.v. injection of interleukin-1β (IL-1β) was unaffected by ovariectomy while the response induced by tumor necrosis factor-α (TNF-α) and macrophage inflammatory protein-1α (MIP-1α) were completely abrogated. These results suggest that the mediators involved in the febrile response in females are similar to males but the reduction of female hormones may decrease the responsiveness of some mediators such as TNF-α and MIP-1α. Compensatory mechanisms may be activated in females after ovariectomy such as an augmented synthesis of COX-2 and PGE2. This article is protected by copyright. All rights reserved.

We compared the structures and rheology of xanthan–galactomannan (X:G) hydrogels with the addition of curcumin in microemulsion (X:GMC) and ethanol (X:GEC). X:GMC hydrogels have gel characteristics and exhibited a significantly higher elastic response than the X:GEC and X:G hydrogels at room temperature, but after heating, an increase in the elastic modulus was observed for the last two systems. The visualization of the hydrogel microstructures by cryo-scanning electronic microscopy revealed pores within the lamellar structure only for X:GMC. In vitro skin permeation tests showed a more pronounced lag time for X:GMC; however, a more efficient permeation from X:GMC than from X:GEC. This study demonstrates that the X:G system is an alternative to traditional gels for the topical applications of hydrophobic drugs. © 2012 Wiley Periodicals, Inc. and the American Pharmacists Association.

Aims Aging and a variety of pathologies, including cancer, diabetes, cardiovascular and inflammatory diseases have been associated with reactive oxygen species (ROS), such as superoxide anion (O2 −), hydroxyl radical ( OH) and hydrogen peroxide (H2O2) generation. Plant polyphenols bear radical scavenging/antioxidant activity. A phytomedicinal preparation obtained from aerial parts of Dicksonia sellowiana (Dicksoniaceae), a native plant from Central and South America, has been widely used in Brazil against asthma and presents beneficial effects in several other diseases, including cardiovascular disturbance. In this work, we investigated whether Dicksonia sellowiana, which is also known to contain high levels of polyphenols, presents antioxidant activity. Methods The antioxidant activity of the hydroalcoholic extract obtained from Dicksonia sellowiana leaves (HEDS) was investigated by in vitro and in vivo tests. Results HEDS (0.1–100 μg/mL) exhibited a strong scavenging activity against all reactive species tested (DPPH, O2 −, OH and H2O2; IC50 = 6.83 ± 2.05, 11.6 ± 5.4, 2.03 ± 0.4, and 4.8 ± 0.4 μg/mL, respectively). HEDS strongly protected endothelial cells against H2O2-induced oxidative stress by mechanisms other than increasing catalase activity. In addition, HEDS protected cell membrane from oxidative damage. HEDS, (20 and 40 mg/kg) inhibited lipid peroxidation in vivo (29.8% and 24.5%, respectively). Conclusions According to our results, we can speculate that the traditional uses of Dicksonia sellowiana for cardiovascular diseases, asthma and skin diseases could be, at least in part, related to the potent antioxidant and endothelial protective activities of the plant.

Correction for ‘Stability of gum Arabic-gold nanoparticles in physiological simulated pHs and their selective effect on cell lines’ by Heloise Ribeiro de Barros et al., RSC Adv., 2016, 6, 9411–9420.

α5β1integrin from both wild-type CHO cells (CHO-K1) and deficient in proteoglycan biosynthesis (CHO-745) is post-translationally modified by glycosaminoglycan chains. We demonstrated this using [35S]sulfate metabolic labeling of the cells, enzymatic degradation, immunoprecipitation reaction with monoclonal antibody, fluorescence microscopy, and flow cytometry. The α5β1integrin heterodimer is a hybrid proteoglycan containing both chondroitin and heparan sulfate chains. Xyloside inhibition of sulfate incorporation into α5β1integrin also supports that integrin is a proteoglycan. Also, cells grown with xyloside adhered on fibronectin with no alteration in α5β1integrin expression. However, haptotactic motility on fibronectin declined in cells grown with xyloside or chlorate as compared with controls. Thus, α5β1integrin is a proteoglycan and the glycosaminoglycan chains of the integrin influence cell motility on fibronectin. Similar glycosylation of α5β1integrin was observed in other normal and malignant cells, suggesting that this modification is conserved and important in the function of this integrin. Therefore, these glycosaminoglycan chains of α5β1integrin are involved in cellular migration on fibronectin.

Endothelins, acting through specific endothelin ET A and/or ET B receptors, participate in nociceptive processing in models of cancer, inflammatory and neuropathic pain. The present study investigated which cell types express endothelin receptors in the trigeminal ganglion, and the contribution of mechanisms mediated by endothelin ET A and ET B receptors to orofacial heat hyperalgesia induced by unilateral constriction of the infraorbital nerve (CION). Both receptor types were identified by immunohistochemistry in the trigeminal ganglion, ET A receptors on small-sized non-myelinated and myelinated A-fibers and ET B receptors on both satellite glial cells and small-sized non-myelinated neuronal cells. CION promoted ipsilateral orofacial heat hyperalgesia which lasted from Day 2 until Day 10 after surgery. Ongoing CION-induced heat hyperalgesia (on Day 4) was reduced transiently, but significantly, by systemic or local treatment with antagonists of endothelin ET A receptors (atrasentan, 10mg/kg, i.v.; or BQ-123, 10nmol/lip), endothelin ET B receptors (A-192621, 20mg/kg, i.v.; or BQ-788, 10nmol/ lip), or of both ET A /ET B receptors (bosentan, 10mg/kg, i.v.; or BQ-123 plus BQ-788, each at 10nmol/lip). On the other hand, CION-induced heat hyperalgesia was transiently abolished over the first 90 min following i.p. injection of morphine hydrochloride (2.5mg/kg), but fully resistant to reversal by indomethacin (4mg/kg, i.p.) or celecoxib (10mg/kg, i.p.). Thus, heat hyperalgesia induced by CION is maintained, in part, by peripheral signaling mechanisms operated by ET A and ET B receptors. Endothelin receptors might represent promising therapeutic targets for the control of trigeminal neuropathic pain.

In vitro, heparin and antithrombotic drugs specifically stimulate the synthesis of an antithrombotic heparan sulfate proteoglycan (HSPG) produced by endothelial cells. The putative heparin binding site(s) that may be related to this phenomenon were investigated. In the preceding article, using various heparin probes, it was shown that the heparin does not bind to the endothelial cell surface, but only to the extracellular matrix. The present study demonstrated that, when the cells were exposed to heparin at 37°C, the heparin was internalized and with time was localized in lysosomes. However, endocytosis of heparin was not required for the stimulation of HSPG synthesis. The requirement for heparin degradation in the stimulus of HSPG synthesis was also investigated. When the cells were incubated with chloroquine, a lysosomotropic amine that raises the lysosomal pH thus inhibiting enzymatic degradation of internalized compounds, stimulation of HSPG synthesis was still observed. These combined results indicate that neither internalization nor degradation of heparin is required for stimulation of HSPG synthesis, and suggests that its binding to the extracellular matrix could be responsible for this effect. J. Cell. Physiol. 217: 360–366, 2008. © 2008 Wiley-Liss, Inc.

Fucan is a term used to denominate a family of sulfated L-fucose-rich polysaccharides. The brown alga Spatoglossum schröederi (Dictyotaceae) has three heterofucans namely fucan A, B and C. The 21 kDa fucan A is composed of a core of a beta (1-3) glucuronic acid-containing oligosaccharide of 4.5 kDa with branches at C4 of the fucose chains alpha (1-3) linked. The fucose is mostly substituted at C4 with a sulfate group and at C2 with chains of beta (1-4) xylose. This fucan has neither anticoagulant (from from 0.1 to 100 microg) nor hemorrhagic activities (from 50 to 800 microg/mL). The antithrombotic test in vivo showed that fucan A has no activity in any of the concentrations (from 0.2 to 20 microg/g/day) tested 1 h after polysaccharide administration. However, when fucan A was injected endovenously 24 h before the ligature of the venae cavae, we observed a dose-dependent effect, reaching saturation at around 20 microg/g of rat weight. In addition, this effect is also time-dependent, reaching saturation around 16 h after fucan administration. In addition, regardless of the administration route, fucan A displayed antithrombotic activity. The exception was the oral pathway. Of particular importance was the finding that fucan A stimulates the synthesis of an antithrombotic heparan sulfate from endothelial cells like heparin. The hypothesis has been raised that the in vivo antithrombotic activity of fucan A is related to the increased production of this heparan. Taken together with the fact that the compound is practically devoid of anticoagulant and hemorrhagic activity, the data suggest that it may be an ideal antithrombotic agent in vivo.

Injuries caused by brown spiders (Loxosceles genus) are associated with dermonecrotic lesions with gravitational spreading and systemic manifestations. The venom has a complex composition containing many different toxins, of which metalloproteases have been described in many different species of this genus. These toxins may degrade extracellular matrix constituents acting as a spreading factor. By using a cDNA library from an Loxosceles intermedia venom gland, we cloned and expressed a 900 bp cDNA, which encoded a signal peptide and a propeptide, which corresponded to a 30 kDa metalloprotease, now named LALP (Loxosceles astacin-like protease). Recombinant LALP was refolded and used to produce a polyclonal antiserum, which showed cross-reactivity with a 29 kDa native venom protein. CD analysis provided evidence that the recombinant LALP toxin was folded correctly, was still in a native conformation and had not aggregated. LALP addition to endothelial cell cultures resulted in de-adhesion of the cells, and also in the degradation of fibronectin and fibrinogen (this could be inhibited by the presence of the bivalent chelator 1,10-phenanthroline) and of gelatin in vitro. Sequence comparison (nucleotide and deduced amino acid), phylogenetic analysis and analysis of the functional recombinant toxin revealed that LALP is related in both structure and function to the astacin family of metalloproteases. This suggests that an astacin-like toxin is present in a animal venom secretion and indicates that recombinant LALP will be a useful tool for future structural and functional studies on venom and the astacin family.