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Accumulation of adipose tissue in specific body areas is related to many physiological and hormonal variables. Spot reduction (SR) is a training protocol aimed to stimulate lipolysis locally, even though this training protocol has not been extensively studied in recent years. Thus, the present study sought to investigate the effect of a circuit-training SR on subcutaneous adipose tissue in healthy adults. Methods: Fourteen volunteers were randomly assigned to spot reduction (SR) or to a traditional resistance training (RT) protocol. Body composition via bioimpedance analysis (BIA) and subcutaneous adipose tissue via skinfold and ultrasound were measured before and after eight weeks of training. Results: SR significantly reduced body mass (p &lt, 0.05) and subcutaneous abdominal adipose tissue (p &lt, 0.05). Conclusions: circuit-training SR may be an efficient strategy to reduce in a localized manner abdominal subcutaneous fat tissue depot.

Considerando que a estrutura vegetativa de especies dos generos brasileiros da subtribo Laeliinae e pouco conhecida, este trabalho objetivou descrever os orgaos vegetativos de Cattleya violacea. Seccoes a mao livre, dos orgaos vegetativos, foram processadas segundo as tecnicas usuais para anatomia vegetal. O pseudobulbo e a folha apresentam hipoderme esclerificada e cuticula espessa. Tais caracteristicas sao adaptacoes ao epifitismo e a necessidade de acumulo de agua. A uniformidade das caracteristicas anatomicas suporta os resultados morfoanatomicos e moleculares para a subtribo Laeliinae (tribo Epidendreae). Caracteres anatomicos e histoquimicos analisados, tais como velame nao distinto em epivelame e endovelame e a presenca de cristais flavonoidicos, apresentam uniformidade generica, o que pode servir como ferramenta para a delimitacao de representantes da subtribo.

Different attentional foci may modify muscle activation during exercises. Our aim was to determine if it is possible to selectively activate the pectoralis major or triceps brachii muscles according to specific verbal instructions provided during the bench press exercise. 13 resistance-trained males (25.6±5.4 yrs, 182.7±9.1 cm, 86.4±9.7 kg) underwent an electromyographic signals acquisition of the sternocostal head, clavicular head of the pectoralis major, the anterior deltoid, and the long head of the triceps brachii (LT) during bench press exercise. Participants performed one non-instructed set (NIS) of 4 repetitions at 50% 1-repetition maximum (1-RM) and one NIS of 4 repetitions at 80% 1-RM. Four additional sets of 4 repetitions at 50% and 80% 1-RM were randomly performed with verbal instructions to isolate the chest muscles (chest instructed set, CIS) or to isolate the triceps muscles (triceps instructed set, TIS). Participants showed significantly higher LT activation during TIS compared to non-instructed set both at 50% (p=0.0199) and 80% 1-RM (p=0.0061) respectively. TIS elicited a significant (p=0.0250) higher activation of LT compared to CIS. Our results suggest that verbal instructions seem to be effective for increasing activity of the triceps brachii but not the pectoralis major during the bench press.

Background p130 Crk-associated substrate (p130CAS; also known as BCAR1) is a scaffold protein that modulates many essential cellular processes such as cell adhesion, proliferation, survival, cell migration, and intracellular signaling. p130Cas has been shown to be highly expressed in a variety of human cancers of epithelial origin. However, few data are available regarding the role of p130Cas during normal epithelial development and homeostasis. Methods To this end, we have generated a genetically modified mouse in which p130Cas protein was specifically ablated in the epidermal tissue. Results By using this murine model, we show that p130Cas loss results in increased cell proliferation and reduction of cell adhesion to extracellular matrix. In addition, epidermal deletion of p130Cas protein leads to premature expression of “late” epidermal differentiation markers, altered membrane E-cadherin/catenin proteins localization and aberrant tyrosine phosphorylation of E-cadherin/catenin complexes. Interestingly, these alterations in adhesive properties in absence of p130Cas correlate with abnormalities in progenitor cells balance resulting in the amplification of a more committed cell population. Conclusion Altogether, these results provide evidence that p130Cas is an important regulator of epidermal cell fate and homeostasis. Electronic supplementary material The online version of this article (10.1186/s12964-018-0289-z) contains supplementary material, which is available to authorized users.

Polyvinylidene difluoride (PVDF) mixed matrix membranes loaded with inorganic TiO2 nanoparticles have received increasing attention in the last few years as self-cleaning membranes for possible application in wastewater treatment and seawater filtration. These novel membranes show increased hydrophilicity, stability and catalytic activity under UV-A irradiation. In this work, PVDF-TiO2 hollow fibers were prepared by employing new strategies for enhancing the stability of the TiO2 dispersion, reducing particle agglomeration and improving their distribution. The spinning conditions for producing ultrafiltration hollow fiber membranes from PVDF material and nano-TiO2 were investigated. Finally, the optimized fibers have been characterized and tested for methylene blue (MB) degradation in water and salty seawater, revealing good permeability, long-term stability under UV-A irradiation, and photo-catalytic activity in both test solutions.

Figure S1. Skin specific deletion of p130Cas/BCAR1 gene. Figure S2. Keratin1 staining. Figure S3. Dot plot quantification of Ki67 and PCNA. Fig. S4 ΔNp63 and YAP expression. Figure S5. Epidermal PCNA, YAP and ΔNp63 expression in WT and p130CasKO mice. Figure S6. Dot plot quantification of loricrin and filaggrin staining. Figure S7. Deletion of p130Cas does not alter expression of β1 and β4 integrins. Figure S8. Epidermal p130Cas loss alters E-cadherin expression and barrier formation. (DOCX 6794 kb)

Published in the book of abstracts of The 11th International Congress on Membranes and Membrane Processes (ICOM 2017). © 2017 Elsevier Limited, Book of poster abstracts 101 to 200 of ICOM 2017, P1.115

STAT3, a pleiotropic transcription factor acting downstream of cytokines and growth factors, is known to enhance proliferation, migration, invasion and aerobic glycolysis in tumors upon aberrant activation. In the murine epidermis, STAT3 is necessary for experimentally induced carcinogenesis. Skin tumorigenesis is conversely enhanced by overexpression in keratinocytes of the constitutively active STAT3C mutant, which also induces robust, psoriasis-like epidermal hyperplasia. We show here that STAT3C expression at physiological levels in knock-in mice leads to mild epidermal hyperplasia and attenuated expression of terminal differentiation markers. Altered differentiation is confirmed in isolated primary epidermal keratinocytes in vitro, correlating with enhanced proliferative and clonogenic potential, attenuated senescence and, strikingly, high-frequency spontaneous immortalization. These results suggest that moderate levels of continuous STAT3 activation, which closely resemble those triggered by chronic inflammation or persistent growth factor stimulation, may establish a preneoplastic state in part by promoting the escape of epidermal progenitor cells from differentiation and senescence checkpoints.

Polyvinylidenefluoride (PVDF) hollow fiber membranes prepared in laboratory through the inversion phase method were characterized and used to clarify an aqueous extract from red wine lees. Steady-state permeate fluxes of 53 kg/m2h were obtained in the treatment of the aqueous extract in selected operating conditions. Suspended solids were completely retained by the hollow fiber membranes while bioactive compounds, including polyphenols, anthocyanins, and resveratrol were recovered in the permeate stream. The clarified stream was then fractionated by nanofiltration (NF). Three different commercial membranes, in flat-sheet configuration (NP010 and NP030 from Microdyn-Nadir, MPF36 from Koch Membrane Systems), were selected and tested for their productivity and selectivity towards sugars and bioactive compounds, including phenolic compounds, anthocyanins, and resveratrol. All selected membranes showed high retention towards anthocyanins (higher than 93%). Therefore, they were considered suitable to concentrate anthocyanins from clarified wine lees extracts at low temperature. On the other hand, NF permeate streams resulted enriched in phenolic compounds and resveratrol. Among the selected membranes, the MPF36 exhibited the lowest retention towards resveratrol (10%) and polyphenols (26.3%) and the best separation factor between these compounds and anthocyanins.

Polyoxometalates (POMs) were used, together with chitosan (CS), to obtain hybrid nanoaggregates. Three representative POMs were efficiently assembled into nanoparticles of few hundred nm diameter, featuring entangled ribbons substructure. In order to establish suitable preparation and stability conditions, the assemblies were characterized in solution by UV-Vis spectroscopy, dynamic light scattering and ζ-potential. The nanoparticles were tested against E. coli (106 CFU/ml) in aqueous solution, showing a synergic activity of the heteropolyacid H5PMo10V2O40 and CS. For such components, a highly porous and antibacterial film was obtained upon lyophilisation of the colloidal mixture., European Union's Seventh Framework Programme (246039); MIUR (FIRB) (prot. RBAP11ETKA)

Noncovalent interactions between the polyoxometalate [PMo12O40](3-) and acryloyloxyundecyltrimethyl ammonium bromide surfactant, used during membrane preparation, were evaluated in the frame of density functional theory. The electronic solvation energy of [PMo12O40](3) and bromide anions was also evaluated, at the same level of theory, in order to predict a probable exchange on the polymeric surface between these anions at the water/polymer interface. Energy balances were theoretically assessed, showing that the bromide cannot be exchanged with this nanosized polyanion in large extent. In order to validate this theoretical conclusion, ad hoc and accurate measurements were carried out by using homemade polymeric membranes and by dipping them in an ca. 0.4 mM solution of Na3[PMo12O40] for 4 days. The Br(-) concentration, released in a polyoxometalate solution, was followed at different times during the test period by gravimetrical analysis. The agreement between the theoretical prediction and experimental data was remarkable, as the quantum calculations correctly accounted for the short-range intermolecular interactions involved in this phenomenon. Bearing in mind that the achieved conclusion is based on an ab initio quantum approach, the findings of this study can be considered rather general and then exploitable for other similar systems.

How metabolic pathways required for epidermal tissue growth and remodeling influence the ability of keratinocytes to survive stressful conditions is still largely unknown. The mechanistic target of rapamycin complex 2 (mTORC2) regulates growth and metabolism of several tissues, but its functions in epidermal cells are poorly defined. Rictor is an adaptor protein essential for mTORC2 activity. To explore the roles of mTORC2 in the epidermis, we have conditionally deleted rictor in mice via K14-Cre-mediated homologous recombination and found that its deficiency causes moderate tissue hypoplasia, reduced keratinocyte proliferation and attenuated hyperplastic response to TPA. Noteworthy, rictor-deficient keratinocytes displayed increased lifespan, protection from senescence, and enhanced tolerance to cellular stressors such as growth factors deprivation, epirubicin and X-ray in vitro and radioresistance in vivo. Rictor-deficient keratinocytes exhibited changes in global gene expression profiles consistent with metabolic alterations and enhanced stress tolerance, a shift in cell catabolic processes from glycids and lipids to glutamine consumption and increased production of mitochondrial reactive oxygen species (ROS). Mechanistically, the resiliency of rictor-deficient epidermal cells relies on these ROS increases, indicating stress resistance via mitohormesis. Thus, our findings reveal a new link between metabolic changes and stress adaptation of keratinocytes centered on mTORC2 activity, with potential implications in skin aging and therapeutic resistance of epithelial tumors.

To elucidate the factors which control the flower phenotypic plasticity of Catasetum fimbriatum, studies focusing the floral development of the species were performed. The treatments were: high light intensity (1613 μmol.m-2.s-1); low light intensity (454 μmol.m-2.s-1) and mean light intensity (827 μmol.m-2.s-1). The floral morphology and anatomy were analyzed to evaluate of the sequence of tissue development involved in the sexual differentiation. We found the presence of cell cluster with meristematic features in the ovaries and gynostemium of the male and female diclinous flowers. These are responsible for the definition of the sexual expression of the species, in the dependence of the environmental stimulus, related to the endogenous concentration of ethylene, allowing, consequently, the dedifferentiation and re-differentiation of those clusters in the sexual determination of the flowers. There were no anatomical differences in floral tissues in different treatments. To elucidate the factors which control the flower phenotypic plasticity of Catasetum fimbriatum, studies focusing the floral development of the species were performed. The treatments were: high light intensity (1613 μmol.m-2.s-1); low light intensity (454 μmol.m-2.s-1) and mean light intensity (827 μmol.m-2.s-1). The floral morphology and anatomy were analyzed to evaluate of the sequence of tissue development involved in the sexual differentiation. We found the presence of cell cluster with meristematic features in the ovaries and gynostemium of the male and female diclinous flowers. These are responsible for the definition of the sexual expression of the species, in the dependence of the environmental stimulus, related to the endogenous concentration of ethylene, allowing, consequently, the dedifferentiation and re-differentiation of those clusters in the sexual determination of the flowers. There were no anatomical differences in floral tissues in different treatments.&nbsp

To elucidate the factors which control the flower phenotypic plasticity of Catasetum fimbriatum, studies focusing the floral development of the species were performed. The treatments were: high light intensity (1613 μmol.m-2.s-1); low light intensity (454 μmol.m-2.s-1) and mean light intensity (827 μmol.m-2.s-1). The floral morphology and anatomy were analyzed to evaluate of the sequence of tissue development involved in the sexual differentiation. We found the presence of cell cluster with meristematic features in the ovaries and gynostemium of the male and female diclinous flowers. These are responsible for the definition of the sexual expression of the species, in the dependence of the environmental stimulus, related to the endogenous concentration of ethylene, allowing, consequently, the dedifferentiation and re-differentiation of those clusters in the sexual determination of the flowers. There were no anatomical differences in floral tissues in different treatments.

Syndecan-4 is a transmembrane heparan sulfate proteoglycan that co-operates with integrins during cell-matrix interactions for the assembly of focal adhesions and actin stress fibers and in the phosphorylation of focal adhesion kinase (FAK) on Tyr397. These cellular events are regulated by the small GTPase Rho, and in the absence of syndecan-4 ligation, cellular levels of GTP-bound Rho are decreased implicating syndecan-4 in the regulation of the small GTPases. In the present study we report that, compared with wild type cells, fibronectin-adherent syndecan-4-null fibroblasts showed enhanced lamellipodia and increased Rac1 activity that could be down-regulated by re-expression of syndecan-4 in the mutant cells. Consistent with the role for Rac1 in activating p38 and JNK signaling, syndecan-4-null cells display higher levels of active p38 MAPK and JNK that were abolished by the expression of a dominant-negative RacN17 mutant. Since p38 and JNK regulate gene expression by phosphorylating and activating transcription factors, we compared both the phosphorylation state and the transcriptional activity of the ATF-2 transcription factor, as a direct p38 and JNK target in syndecan-4-null and wild type cells. In the absence of syndecan-4, both ATF-2 phosphorylation and transcriptional activity were significantly more elevated compared with wild type cells, and both activities were decreased either by the re-expression of syndecan-4 or by the expression of RacN17. Our results reveal a novel function for syndecan-4 in modulating nuclear transcriptional activity and indicate an underlying mechanism that acts at the level of Rac1-p38/JNK signaling.

Syndecan-4 and integrins are the primary transmembrane receptors of focal adhesions in cells adherent to extracellular matrix molecules. Syndesmos is a cytoplasmic protein that interacts specifically with the cytoplasmic domain of syndecan-4, and it co-localizes with syndecan-4 in focal contacts. In the present study we sought possible interactors with syndesmos. We find that syndesmos interacts with the focal adhesion adaptor protein paxillin. The binding of syndesmos to paxillin is direct, and these interactions are triggered by the activation of protein kinase C. Syndesmos also binds the paxillin homolog, Hic-5. The connection of syndecan-4 with paxillin through syndesmos parallels the connection between paxillin and integrins and may thus reflect the cooperative signaling of these two receptors in the assembly of focal adhesions and actin stress fibers.

The syndecans make up a family of transmembrane heparan sulfate proteoglycans that act as coreceptors with integrins and growth factor tyrosine kinase receptors. Syndecan-4 is upregulated in skin dermis after wounding, and, in cultured fibroblasts adherent to the ECM protein fibronectin, this proteoglycan signals cooperatively with beta1 integrins. In this study, we generated mice in which the syndecan-4 gene was disrupted by homologous recombination in embryonic stem cells to test the hypothesis that syndecan-4 contributes to wound repair. Mice heterozygous or homozygous for the disrupted syndecan-4 gene are viable, fertile, and macroscopically indistinguishable from wild-type littermates. Compared with wild-type littermates, mice heterozygous or homozygous for the disrupted gene have statistically significant delayed healing of skin wounds and impaired angiogenesis in the granulation tissue. These results indicate that syndecan-4 is an important cell-surface receptor in wound healing and angiogenesis and that syndecan-4 is haplo-insufficient in these processes.

An algal biosensor for toxicity assessment of estuarine waters is proposed. The sensor was obtained by coupling a suited algal bioreceptor (the cyanobacterium Spirulina subsalsa) to an amperometric gas diffusion electrode. The analytical device allows the monitoring of the evolution of photosynthetic O2 and the detection of alterations due to toxic effects caused by environmental pollutants present in the medium. Four chemical species representative of three main different classes of pollutants (heavy metals, triazinic herbicides, carbamate insecticides) were tested at different concentrations using a standardized natural water as experimental medium. In all the cases a toxic response was detected (i.e. a dose-related inhibition of photosynthetic activity was recorded) with good reproducibility.

Syndecan-4 is a cell surface heparan sulfate proteoglycan which, in cooperation with integrins, transduces signals for the assembly of focal adhesions and actin stress fibers in cells plated on fibronectin. The regulation of these cellular events is proposed to occur, in part, through the interaction of the cytoplasmic domains of these transmembrane receptors with intracellular proteins. To identify potential intracellular proteins that interact with the cytoplasmic domain of syndecan-4, we carried out a yeast two-hybrid screen in which the cytoplasmic domain of syndecan-4 was used as bait. As a result of this screen, we have identified a novel cellular protein that interacts with the cytoplasmic domain of syndecan-4 but not with those of the other three syndecan family members. The interaction involves both the membrane proximal and variable central regions of the cytoplasmic domain. We have named this cDNA and encoded protein syndesmos. Syndesmos is ubiquitously expressed and can be myristylated. Consistent with its myristylation and syndecan-4 association, syndesmos colocalizes with syndecan-4 in the ventral plasma membranes of cells plated on fibronectin. When overexpressed in NIH 3T3 cells, syndesmos enhances cell spreading, actin stress fiber and focal contact formation in a serum-independent manner.

The formation of focal adhesions and actin stress fibers on fibronectin is dependent on signaling through (β)1 integrins and the heparan sulfate proteoglycan syndecan-4, and we have analyzed the requirement of the glycosaminoglycan chains of syndecan-4 during these events. Chinese hamster ovary cells with mutations in key enzymes of the glycanation process do not synthesize glycosaminoglycan chains and are unable to assemble actin stress fibers and focal contacts when cultured on fibronectin. Transfection of the mutant cells with a cDNA that encodes the core protein of chicken syndecan-4 leads to the production of unglycanated core protein. The overexpression of syndecan-4 core protein in these mutant cells increases cell spreading and is sufficient for these cells to assemble actin stress fibers and focal adhesions similar to wild-type cells seeded on fibronectin and vitronectin matrices. Syndecan-4 core protein colocalizes to focal contacts in mutant cells that have been transfected with the syndecan-4 core protein cDNA. These data indicate an essential role for the core protein of syndecan-4 in the generation of signals leading to actin stress fiber and focal contact assembly.

The assembly of focal adhesions and actin stress fibers by cells plated on fibronectin depends on adhesion-mediated signals involving both integrins and cell-surface heparan sulfate proteoglycans. These two cell-surface receptors interact with different domains of fibronectin. To attempt to identify the heparan sulfate proteoglycans involved, we used fibronectin-null (FN−/−) mouse fibroblasts to eliminate the contribution of endogenous fibronectin during the analysis. FN−/− fibroblasts plated on the cell-binding domain of fibronectin or on antibodies directed against mouse β1 integrin chains attach but fail to spread and do not form focal adhesions or actin stress fibers. When such cells are treated with antibodies directed against the ectodomain of mouse syndecan-4, they spread fully and assemble focal adhesions and actin stress fibers indistinguishable from those seen in cells plated on intact fibronectin. These results identify syndecan-4 as a heparan sulfate proteoglycan involved in the assembly process. The antibody-stimulated assembly of focal adhesions and actin stress fibers in cells plated on the cell-binding domain of fibronectin can be blocked with C3 exotransferase, an inhibitor of the small GTP-binding protein Rho. Treatment of cells with lysophosphatidic acid, which activates Rho, results in full spreading and assembly of focal adhesions and actin stress fibers in fibroblasts plated on the cell-binding domain of fibronectin. We conclude that syndecan-4 and integrins can act cooperatively in generating signals for cell spreading and for the assembly of focal adhesions and actin stress fibers. We conclude further that these joint signals are regulated in a Rho-dependent manner.

Signals downstream of Akt can either favor or oppose stem cell (SC) maintenance, but how this dual role can be achieved is still undefined. Using human limbal keratinocyte stem cells (LKSCs), a SC type used in transplantation therapies for corneal regeneration, we show that Akt signaling is prominent in SC populations both in vivo and in vitro, and that Akt1 promotes while Akt2 opposes SC self-renewal. Noteworthy, loss of Akt2 signaling enhances LKSC maintenance ex vivo, whereas Akt1 depletion anticipates SC exhaustion. Mechanistically, the antagonistic functions of Akt1 and Akt2 in SC control are mainly dictated by their differential subcellular distribution, being nuclear Akt2 selectively implicated in FOXO inhibition. Akt2 downregulation favors LKSC maintenance as a result of a gain of FOXO functions, which attenuates the mechanistic target of rapamycin complex one signaling via tuberous sclerosis one gene induction, and promotes growth factor signaling through Akt1. Consistently, Akt2 deficiency also enhances limbal SCs in vivo. Thus, our findings reveal distinct roles for nuclear versus cytosolic Akt signaling in normal epithelial SC control and suggest that the selective Akt2 inhibition may provide novel pharmacological strategies for human LKSC expansion in therapeutic settings and mechanistic research. Stem Cells 2014;32:754–769