High lipid content of irradiated human melanoma cells does not affect cytokine-matured dendritic cell function

Gamma irradiation is one of the methods used to sterilize melanoma cells prior to coculturing them with monocyte-derived immature dendritic cells in order to develop antitumor vaccines. However, the changes taking place in tumor cells after irradiation and their interaction with dendritic cells have been scarcely analyzed. We demonstrate here for the first time that after irradiation a fraction of tumor cells present large lipid bodies, which mainly contain triglycerides that are several-fold increased as compared to viable cells as determined by staining with Oil Red O and BODIPY 493/503 and by biochemical analysis. Phosphatidyl-choline, phosphatidyl-ethanolamine and sphingomyelin are also increased in the lipid bodies of irradiated cells. Lipid bodies do not contain the melanoma-associated antigen MART-1. After coculturing immature dendritic cells with irradiated melanoma cells, tumor cells tend to form clumps to which dendritic cells adhere. Under such conditions, dendritic cells are unable to act as stimulating cells in a mixed leukocyte reaction. However, when a maturation cocktail composed of TNF-alpha, IL-6, IL-1beta and prostaglandin E2 is added to the coculture, the tumor cells clumps disaggregate, dendritic cells remain free in suspension and their ability to efficiently stimulate allogeneic lymphocytes is restored. These results help to understand the events following melanoma cell irradiation, shed light about interactions between irradiated cells and dendritic cells, and may help to develop optimized dendritic cell vaccines for cancer therapy.     

A Pseudopterane Diterpene Isolated From the Octocoral Pseudopterogorgia acerosa Inhibits the Inflammatory Response Mediated by TLR-Ligands and TNF-Alpha in Macrophages

Several diterpenoids isolated from terrestrial and marine environments have been identified as important anti-inflammatory agents. Although considerable progress has been made in the area of anti-inflammatory treatment, the search for more effective and safer compounds is a very active field of research. In this study we investigated the anti-inflammatory effects of a known pseudopterane diterpene (referred here as compound 1) isolated from the octocoral Pseudopterogorgia acerosa on the tumor necrosis factor- alpha (TNF-α) and TLRs- induced response in macrophages. Compound 1 inhibited the expression and secretion of the inflammatory mediators TNF-α, interleukin (IL)-6, IL-1β, nitric oxide (NO), interferon gamma-induced protein 10 (IP-10), ciclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) and monocyte chemoattractant protein-1 (MCP-1) induced by LPS in primary murine macrophages. This effect was associated with the inhibition of IκBα degradation and subsequent activation of NFκB. Compound 1 also inhibited the expression of the co-stimulatory molecules CD80 and CD86, which is a hallmark of macrophage activation and consequent initiation of an adaptive immune response. The anti-inflammatory effect was not exclusive to LPS because compound 1 also inhibited the response of macrophages to TNF-α and TLR2 and TLR3 ligands. Taken together, these results indicate that compound 1 is an anti-inflammatory molecule, which modulates a variety of processes occurring in macrophage activation.     

Effects of inoculum concentration,temperature, and carbon sources on tannase production during solid state fermentation of cashew apple bagasse

In this study, the optimization of tannase production by solid state fermentation was investigated using cashew apple bagasse (CAB), an inexpensive residue produced by the cashew apple agroindustry, as a substrate. To accomplish this, CAB was enriched with 2.5% (w/w) tannic acid and 2.5% (w/w) ammonium sulphate and then moistened with water (60 mL/100 g of dry CAB). The influence of inoculum concentration (10⁴ to 10⁷ spores/g), temperature (20, 25, 30, and 35°C) and several additional carbon sources (glucose, starch, sucrose, maltose, analytical grade glycerol, and glycerol produced during biodiesel production) on enzyme production by Aspergillus oryzae was then evaluated. Supplementation with maltose and glycerol inhibited tannase synthesis, which resulted in lower enzyme activity. Starch and sucrose supplementation increased enzyme production, but decreased the enzyme productivity. The maximum tannase activity (4.63 units/g of dry substrate) was obtained at 30°C, using 10⁷ spores/g and 1.0% (w/v) sucrose as an additional carbon source.     

Addition of α-O-GlcNAc to threonine residues define the post-translational modification of mucin-like molecules in Trypanosoma cruzi

Trypanosoma cruzi, an intracellular protozoan etiologic agent of Chagas disease is covered by a dense coat of mucin-type glycoproteins, which is important to promote the parasite entry and persistence in the mammalian host cells. The O-glycosylation of T. cruzi mucins (Tc-mucins) is initiated by enzymatic addition of α-O-N-acetylglucosamine (GlcNAc) to threonine (Thr) by the UDP-GlcNAc:polypeptide α-N-acetylglucosaminyltransferase (pp-α-GlcNAcT) in the Golgi. The Tc-mucin is characterized by the presence of a high structural diversity of O-linked oligosaccharides found among different parasite strains, comprising two O-glycan Cores. In the Core 1, from strains principally associated with the domestic transmission cycle of Chagas disease, the GlcNAc O-4 is substituted with a β-galactopyranose (βGalp) unit, and in the most complex oligosaccharides the GlcNAc O-6 is further processed by the addition of β1?→?2-linked Galp residues creating a short linear Galp-containing chain. In the Core 2 structures, expressed by strains isolated from T. cruzi sylvatic hosts, the GlcNAc O-4 carries a β-galactofuranose (βGalf) unit and the GlcNAc O-6 can carry a branched Galpβ1?→?3[Galpβ1?→?2]Galpβ1?→?6 motif. The O-glycans carrying nonreducing terminal βGalp are available for sialylation by a surface T. cruzi trans-sialidase activity. Based on structural results, this review summarizes available data on the highly conserved process, which adds the GlcNAc unit in α-linkage to Thr residues the basis of the post-translational modification system in T. cruzi mucins. In addition, a mechanism unique employed by the parasite to transfer exogenous sialic acid residues to Tc-mucins is presented.     

Activation of Neck and Low-Back Muscles Is Reduced with the Use of a Neck Balance System Together with a Lumbar Support in Urban Drivers

Driving is associated with high activation of low-back and neck muscles due to the sitting position and perturbations imposed by the vehicle. The aim of this study was to investigate the use of a neck balance system together with a lumbar support on the activation of low-back and neck muscles during driving. Twelve healthy male subjects (age 32±6.71 years) were asked to drive in two conditions: 1) with devices; 2) without devices. During vehicle accelerations and decelerations root mean square (RMS) of surface electromyography (sEMG) was recorded from the erector spinae, semispinalis capitis and sternocleidomastoid muscles and expressed as a percentage of maximal voluntary contraction (MVC). The pitch of the head was obtained by means of an inertial sensor placed on the subjects’ head. A visual analog scale (VAS) was used to assess the level of perceived comfort. RMS of the low back muscles was lower with than without devices during both acceleration and deceleration of the vehicle (1.40±0.93% vs 29 2.32±1.90% and 1.88±1.45% vs 2.91±2.33%, respectively), while RMS of neck extensor muscles was reduced only during acceleration (5.18±1.96% vs 5.91±2.16%). There were no differences between the two conditions in RMS of neck flexor muscles, the pitch of the head and the VAS score. The use of these two ergonomic devices is therefore effective in reducing the activation of low-back and neck muscles during driving with no changes in the level of perceived comfort, which is likely due to rebalancing weight on the neck and giving a neutral position to lumbar segments.     

Cytoskeleton structure,pattern of mitochondrial activity and ultrastructure of frozen or vitrified sheep embryos

Even though sheep embryo cryopreservation is a commonly used procedure the survival and pregnancy outcomes can vary greatly. This study investigated whether cryopreservation was causing subtle changes in ultrastructure, mitochondrial activity or cytoskeletal integrity. Sheep embryos were either slow cooled in 1.5 M EG (n = 22), or vitrified in 20% EG + 20% DMSO with 0.5 M sucrose in Open Pulled Straws (OPS) (n = 24). One hour after warming the cryopreserved embryos differed from control embryos in that they had no mitochondrial activity combined with cytoskeletal disorganization and large vesicles. Vitrified embryos also showed many points of cytoskeleton disruption. Ultrastructural alterations resulting from actin filaments disorganization were observed in both cryopreserved groups. This includes areas presenting no cytoplasmic organelles, Golgi complex located far from the nucleus and a decrease of specialized intercellular junctions. Additionally, large vesicles were observed in vitrified morulae and early blastocysts. The alterations after cryopreservation were proportional to embryo quality as assessed using the stereomicroscope. Even in the absence of mitochondrial activity, grade I and II cryopreserved embryos contained mitochondria with normal ultrastructure. Embryos classified as grade I or II in the stereomicroscope revealed mild ultrastructural alterations, meaning that this tool is efficient to evaluate embryos after cryopreservation.     

Limosilactobacillus fermentum,Current Evidence on the Antioxidant Properties and Opportunities to be Exploited as a Probiotic Microorganism

Probiotics and Antimicrobial Proteins - The unbalance in the production and removal of oxygen-reactive species in the human organism leads to oxidative stress, a physiological condition commonly...     

Dose measurements in a thorax phantom at 3DCRT breast radiation therapy

BackgroundThe anthropomorphic and anthropometric phantom developed by the research group NRI (Núcleo de Radiações Ionizantes) can reproduce the effects of the interactions of radiation occurring in the human body. The whole internal radiation transport phenomena can be depicted by film dosimeters in breast RT. Our goal was to provide a dosimetric comparison of a radiation therapy (RT) plan in a 4MV 3D-conformal RT (4MV-3DCR T) and experimental data measured in a breast phantom.Materials and methodsThe RT modality was two parallel opposing fields for the left breast with a prescribed dose of 2.0 Gy in 25 fractions. The therapy planning system (TPS) was performed on CA T3D software. The dose readings at points of interest (POI) pre-established in TPS were recorded. An anthropometric thorax-phantom with removal breast was used. EBT2 radiochromic films were inserted into the ipisilateral breast, contralateral breast, lungs, heart and skin. The irradiation was carried out on 4/80 Varian linear accelerator at 4MV.ResultsThe mean dose at the OAR’s presented statistically significant differences (p < 0.001) of 34.24%, 37.96% and 63.47% for ipsilateral lung, contralateral lung, and heart, respectively. The films placed at the skin-surface interface in the ipsilateral breast also showed statistically significant differences (p < 0.001) of 16.43%, −10.16%, −14.79% and 15.67% in the four quadrants, respectively. In contrast, the PTV dosimeters, representative of the left breast volume, encompassed by the electronic equilibrium, presented a non-significant difference with TPS, p = 0.20 and p = 0.90.ConclusionThere was a non-significant difference of doses in PTV with electronic equilibrium; although no match is achieved outside electronic equilibrium.     

Bivariate GWAS reveals pleiotropic regions among feed efficiency and beef quality-related traits in Nelore cattle

Mammalian Genome - Feed-efficient cattle selection is among the most leading solutions to reduce cost for beef cattle production. However, technical difficulties in measuring feed efficiency traits...