Caffeic acid derivatives: in vitro and in vivo anti-inflammatory properties

Caffeic acid and some of its derivatives such as caffeic acid phenetyl ester (CAPE) and octyl caffeate are potent antioxidants which present important anti-inflammatory actions. The present study assessed the in vitro and in vivo effects of five caffeic acid derivatives (caffeic acid methyl, ethyl, butyl, octyl and benzyl esters) and compared their actions to those of CAPE. In the model of LPS-induced nitric oxide (NO) production in RAW 264.7 macrophages, the pre-incubation of all derivatives inhibited nitrite accumulation on the supernatant of stimulated cells, with mean IC₅₀ (μM) values of 21.0, 12.0, 8.4, 2.4, 10.7 and 4.80 for methyl, ethyl, butyl, octyl, benzyl and CAPE, respectively. The effects of caffeic acid derivatives seem to be related to the scavenging of NO, as the compounds prevented SNAP-derived nitrite accumulation and decreased iNOS expression. In addition, butyl, octyl and CAPE derivatives significantly inhibited LPS-induced iNOS expression in RAW 264.7 macrophages. Extending the in vitro results, we showed that the pre-treatment of mice with butyl, octyl and CAPE derivatives inhibited carrageenan-induced paw edema and prevented the increase in IL-1β levels in the mouse paw by 30, 24 and 36%, respectively. Butyl, octyl and CAPE derivatives also prevented carrageenan-induced neutrophil influx in the mouse paw by 28, 49 and 31%, respectively. Present results confirm and extend literature data, showing that caffeic acid derivatives exert in vitro and in vivo anti-inflammatory actions, being their actions mediated, at least in part by the scavenging of NO and their ability to modulate iNOS expression and probably that of other inflammatory mediators.     

Cytotoxic,hypoglycemic activity and phytochemical analysis of Rubus imperialis (Rosaceae)

Rubus imperialis, Artemia salina, 3-O-methylellagic-4'-O-alpha-rhamnose Acid Screening of different extracts, fractions and compounds from Rubus imperialis Chum. Schl. (Rosaceae) has been conduced using the brine shrimp microwell cytotoxicity assay. Three parts of the plant (methanolic extract from leaves, roots and stems), three fractions from roots (hexane, ethyl acetate and butanol) and three isolated compounds (niga-ichigoside F1, 23-hydroxytormentic acid, ellagic acid derivative) were tested. The most promising material (LC50 <1000 microg/ml) were the methanolic extract and ethyl acetate fraction from roots. However, there was little correlation observed in the degree of toxicities observed between the isolated compounds. On the other hand, the cytotoxicity and in vivo assays confirmed the hypoglycemic activity of methanolic extract and validated the Brazilian popular use of R. imperialis as an antidiabetic agent.     

Evidence That Lipopolisaccharide May Contribute to the Cytokine Storm and Cellular Activation in Patients with Visceral Leishmaniasis

Background

Visceral leishmaniasis (VL) is characterized by parasite-specific immunosuppression besides an intense pro-inflammatory response. Lipopolisaccharide (LPS) has been implicated in the immune activation of T-cell deficient diseases such as HIV/AIDS and idiopathic lymphocytopenia. The source of LPS is gram-negative bacteria that enter the circulation because of immunological mucosal barrier breakdown. As gut parasitization also occurs in VL, it was hypothesized that LPS may be elevated in leishmaniasis, contributing to cell activation.

Methodology/Principal Findings

Flow cytometry analysis and immunoassays (ELISA and luminex micro-beads system) were used to quantify T-cells and soluble factors. Higher LPS and soluble CD14 levels were observed in active VL in comparison to healthy subjects, indicating that LPS was bioactive; there was a positive correlation between these molecules (r = 0.61;p<0.05). Interestingly, LPS was negatively correlated with CD4⁺ (r = −0.71;p<0.01) and CD8⁺ T-cells (r = −0.65;p<0.05). Moreover, higher levels of activation-associated molecules (HLA-DR, CD38, CD25) were seen on T lymphocytes, which were positively associated with LPS levels. Pro-inflammatory cytokines and macrophage migration inhibitory factor (MIF) were also augmented in VL patients. Consistent with the higher immune activation status, LPS levels were positively correlated with the inflammatory cytokines IL-6 (r = 0.63;p<0.05), IL-8 (r = 0.89;p<0.05), and MIF (r = 0.64;p<0.05). Also, higher plasma intestinal fatty acid binding protein (IFABP) levels were observed in VL patients, which correlated with LPS levels (r = 0.57;p<0.05).

Conclusions/Significance

Elevated levels of LPS in VL, in correlation with T-cell activation and elevated pro-inflammatory cytokines and MIF indicate that this bacterial product may contribute to the impairment in immune effector function. The cytokine storm and chronic immune hyperactivation status may contribute to the observed T-cell depletion. LPS probably originates from microbial translocation as suggested by IFABP levels and, along with Leishmania antigen-mediated immune suppression, may play a role in the immunopathogenesis of VL. These findings point to possible benefits of antimicrobial prophylaxis in conjunction with anti-Leishmania therapy.     

Implant-supported palatal lift prosthesis in a patient with velopharyngeal incompetence: a case report

doi: 10.1111/j.1741‐2358.2011.00561.x Implant‐supported palatal lift prosthesis in a patient with velopharyngeal incompetence: a case report Objective: To describe the use of dental implants in the treatment of velopharyngeal incompetence. Background: Velopharyngeal incompetence is characterized by the inability to contract the tissues of the soft palate. The most common causes are neuromuscular problems resulting from concussion and degenerative conditions of the central nervous system. Materials and methods: The treatment using palatal lift prosthesis is well established in the literature; however, reports on the use of osseointegrated implants are scarce. Rehabilitation using only this type of fibromucosal support is difficult in edentulous patients, since the prosthesis has a palatal lift extension. Results: The implants provided retention and stability to the prosthesis, improving swallowing and speech and contributing to improve the patient’s quality of life. Conclusion: This article reports the rehabilitation of a patient with a totally edentulous maxilla with velopharyngeal incompetence using an implant‐retained palatal lift prosthesis.     

Regulation of inflammatory chemokine receptors on blood T cells associated to the circulating versus liver chemokines in dengue fever

Little is known about the role of chemokines/chemokines receptors on T cells in natural DENV infection. Patients from DENV-2 and -3- outbreaks were studied prospectively during the acute or convalescent phases. Expression of chemokine receptor and activation markers on lymphocyte subpopulations were determined by flow cytometry analysis, plasma chemokine ligands concentrations were measured by ELISA and quantification of CCL5/RANTES(+) cells in liver tissues from fatal dengue cases was performed by immunochemistry. In the acute DENV-infection, T-helper/T-cytotoxic type-1 cell (Th1/Tc1)-related CCR5 is significantly higher expressed on both CD4 and CD8 T cells. The Th1-related CXCR3 is up-regulated among CD4 T cells and Tc2-related CCR4 is up-regulated among CD8 T cells. In the convalescent phase, all chemokine receptor or chemokine ligand expression tends to reestablish control healthy levels. Increased CCL2/MCP-1 and CCL4/MIP-1β but decreased CCL5/RANTES levels were observed in DENV-patients during acute infection. Moreover, we showed an increased CD107a expression on CCR5 or CXCR3-expressing T cells and higher expression of CD29, CD44(HIGH) and CD127(LOW) markers on CCR4-expressing CD8 T cells in DENV-patients when compared to controls. Finally, liver from dengue fatal patients showed increased number of cells expressing CCL5/RANTES in three out of four cases compared to three death from a non-dengue patient. In conclusion, both Th1-related CCR5 and CXCR3 among CD4 T cells have a potential ability to exert cytotoxicity function. Moreover, Tc1-related CCR5 and Tc2-related CCR4 among CD8 T cells have a potential ability to exert effector function and migration based on cell markers evaluated. The CCR5 expression would be promoting an enhanced T cell recruitment into liver, a hypothesis that is corroborated by the CCL5/RANTES increase detected in hepatic tissue from dengue fatal cases. The balance between protective and pathogenic immune response mediated by chemokines during dengue fever will be discussed.     

Collagen peptides modulate the metabolism of extracellular matrix by human dermal fibroblasts derived from sun‐protected and sun‐exposed body sites

Clinical data published in recent years have demonstrated positive effects of collagen hydrolysate (CH) on skin aging clinical signs. CH use as food supplement has a long history; however, few studies have addressed the underlying purpose of CH on the cellular and molecular biology of skin cells that could elucidate clinical improvement findings. Wide diversity of characteristics has been reported for dermal fibroblasts derived from different body sites and it is unknown whether collagen peptides could modulate differently cells from chronological aged and photoaged skin areas. This study investigated the influence of CH on the extracellular matrix metabolism and proliferation of human dermal fibroblasts (HDFs) derived from chronological aged (sun‐protected) and photoaged (sun‐exposed) body sites. CH treatment did not affect cellular proliferation of either cell cultures, but notably modulated cell metabolism in monolayer model, increasing the content of dermal matrix precursor and main protein, procollagen I and collagen I, respectively. These effects were confirmed in the human dermal equivalent model. The increase in collagen content in the cultures was attributed to stimulation of biosynthesis and decreased collagen I metabolism through inhibition of metalloproteinase activity (MMP) 1 and 2. Modulation of CH in dermal metabolism did not differ between cells derived from sun‐protected and sun‐exposed areas, although lower concentrations of CH seemed to be enough to stimulate sun‐exposed‐derived HDFs, suggesting more pronounced effect in these cells. This study contributes to understanding the biological effects of CH on skin cells and viability of its use as a functional ingredient in food supplements.     

Factors affecting the structure of Coleoptera assemblages on bracket fungi (Basidiomycota) in a Brazilian forest

Insect–fungal interactions are an important but understudied aspect of tropical forest ecology. Here we present the first large‐scale study of insect communities feeding on the reproductive structures of macrofungi (basidiomes) in the Neotropics. This trophic interaction is not well characterized in most ecosystems; however, beetle consumption of basidiomes is thought to be affected by fungal factors, via mechanisms analogous to those observed in plant–herbivore interactions and in some interactions with fungi as hosts in the Holarctic region. We investigated how the composition of beetle assemblages varies as a function of fungal taxonomic distance, basidiome consistency, and hyphal systems. We collected 367 basidiomes belonging to the orders Polyporales and Hymenochaetales in the subtropical Araucaria angustifolia forest region of southern Brazil, along with any fauna present or without it. Basidiomes were maintained individually in the laboratory in plastic containers for up to three months to allow beetles to develop to adulthood, at which point the beetles were collected. We found that 207 basidiome specimens representing 40 species were associated with beetles. We recorded 447 occurrences of Coleoptera, representing 90 morphospecies from 20 families. We found that assemblages of fungivorous Coleoptera were more similar among more closely related fungi. Furthermore, the beetle assemblages varied as a function of basidiome toughness, which is influenced by sporocarp consistency and hyphal system type. The associations between beetles and basidiomes resemble those reported previously in temperate zones, suggesting continuity in the structure of such associations across a wide latitudinal range.     

How can a life cycle inventory parametric model streamline life cycle assessment in the wooden pallet sector?

Purpose

This study discusses the use of parameterization within the life cycle inventory (LCI) in the wooden pallet sector, in order to test the effectiveness of LCI parametric models to calculate the environmental impacts of similar products. Starting from a single case study, the objectives of this paper are (1) to develop a LCI parametric model adaptable to a range of wooden pallets, (2) to test this model with a reference product (non-reversible pallet with four-way blocks) and (3) to determine numerical correlations between the environmental impacts and the most significant LCI parameters; these correlations can be used to improve the design of new wooden pallets.

Methods

The conceptual scheme for defining the model is based on ISO14040-44 standards. First of all, the product system was defined identifying the life cycle of a generic wood pallet, as well as its life cycle stages. A list of independent and dependent parameters was used to describe the LCI flows of a generic wooden pallet. The LCI parametric model was applied to calculate the environmental impacts of the reference product, with regard to a selection of impact categories at midpoint level (climate change, human toxicity, particulate matter formation, agricultural land occupation, fossil depletion). The model was then applied to further 11 wooden pallets belonging to the same category.

Results and discussion

The definition of a LCI parametric model based on 31 independent parameters and 21 dependent parameters streamlined the data collection process, as the information required for fulfilling the LCI are standard information about the features of the wooden pallet and its manufacturing process. The contribution analysis on the reference product revealed that the most contributing life cycle stages are wood and nails extraction and manufacturing (positive value of environmental impact) and end-of-life (avoided impact). This result is driven by two parameters: mass of wood and average distance for transport of wood. Based on the results of the application of the LCI parametric model to the identified products, one parameter-based regression and one multiple non-linear regression allowed to define a correlation between the life cycle impact assessment (LCIA) category indicators considered and the most influencing parameters.

Conclusions

The definition of LCI parametric model in the wooden pallet sector can effectively be used for calculating the environmental impacts of products with different designs, as well as for obtaining a preliminary estimation of the life cycle environmental impacts of new products.