PIP3-dependent macropinocytosis is incompatible with chemotaxis

In eukaryotic chemotaxis, the mechanisms connecting external signals to the motile apparatus remain unclear. The role of the lipid phosphatidylinositol 3,4,5-trisphosphate (PIP3) has been particularly controversial. PIP3 has many cellular roles, notably in growth control and macropinocytosis as well as cell motility. Here we show that PIP3 is not only unnecessary for Dictyostelium discoideum to migrate toward folate, but actively inhibits chemotaxis. We find that macropinosomes, but not pseudopods, in growing cells are dependent on PIP3. PIP3 patches in these cells show no directional bias, and overall only PIP3-free pseudopods orient up-gradient. The pseudopod driver suppressor of cAR mutations (SCAR)/WASP and verprolin homologue (WAVE) is not recruited to the center of PIP3 patches, just the edges, where it causes macropinosome formation. Wild-type cells, unlike the widely used axenic mutants, show little macropinocytosis and few large PIP3 patches, but migrate more efficiently toward folate. Tellingly, folate chemotaxis in axenic cells is rescued by knocking out phosphatidylinositide 3-kinases (PI 3-kinases). Thus PIP3 promotes macropinocytosis and interferes with pseudopod orientation during chemotaxis of growing cells

Capacitação em agroecologia na comunidade quilombola do Varzeão, Dr. Ulysses, Vale do Ribeira, PR.

O projeto "Assistência Técnica e Extensão Rural - ATER - e Capacitação em Agroecologia na Comunidade Quilombola do Varzeão, Dr. Ulysses, Vale do Ribeira" atua desde 2007 e procura, através da Agroecologia e dos seus princípios, contribuir com a organização das comunidades, estimulando o redesenho dos agroecossistemas. Os Quilombolas têm uma produção moldada através de métodos tradicionais e estão próximos dos princípios da Agroecologia. A comunidade remanescente de Quilombo do Varzeão se localiza em Dr. Ulysses e conta hoje com 19 famílias com um total de 56 pessoas com diversos problemas no que tange à sua situação social. O projeto surgiu por meio de contatos com as diversas organizações que atuam no Vale do Ribeira, isso desembocou em uma aproximação com a comunidade do Varzeão. Dessa forma, discutiu-se com a comunidade as demandas para um projeto de extensão, o que culminou na aprovação do projeto junto a Secretaria de Ciência e Tecnologia do Ensino Superior (SETI) - Universidade Sem Fronteiras e a Universidade Federal do Paraná (UFPR).Disponível também em: Cadernos de Agroecologia, V. 5, n.1, 2010

Intratumoural immunotherapies for unresectable and metastatic melanoma: current status and future perspectives.

The emergence of human intratumoural immunotherapy (HIT-IT) is a major step forward in the management of unresectable melanoma. The direct injection of treatments into melanoma lesions can cause cell lysis and induce a local immune response, and might be associated with a systemic immune response. Directly injecting immunotherapies into tumours achieves a high local concentration of immunostimulatory agent while minimising systemic exposure and, as such, HIT-IT agents are associated with lower toxicity than systemic immune checkpoint inhibitors (CPIs), enabling their potential use in combination with other therapies. Consequently, multiple HIT-IT agents, including oncolytic viruses, pattern-recognition receptor agonists, injected CPIs, cytokines and immune glycolipids, are under investigation. This review considers the current clinical development status of HIT-IT agents as monotherapy and in combination with systemic CPIs, and the practical aspects of administering and assessing the response to these agents. The future of HIT-IT probably lies in its use in combination with systemic CPIs; data from Phase 2 trials indicate a synergy between HIT-IT and CPIs. Data also suggest that the addition of HIT-IT to a CPI might generate responses in CPI-refractory tumours, thereby overcoming resistance and addressing a current unmet need in unresectable and metastatic melanoma for treatment options following progression after CPI treatment

Teor de óleo essencial de Ocimum selloi Benth. sob diferentes doses de compostos orgânicos.

Em espécies bioativas, a aplicação de compostos orgânicos tem sido relacionada à produção de metabólitos secundários. No presente estudo, avaliou-se o efeito de diferentes doses de compostos orgânicos no teor de óleo essencial de alfavaca anisada, fornecendo bases científicas para agricultura orgânica. Em Curitiba-PR, ano de 2008, num delineamento em blocos ao acaso, com três repetições, doses (0, 0,85, 1,7 e 3 t.ha-1) de compostos estabilizados (A, B, C e D), elaborados com diferentes materiais orgânicos, foram utilizadas no cultivo de alfavaca anisada. O espaçamento utilizado em cultivo protegido irrigado foi de 0,5m entre linhas por 1,0m entre plantas. Aos seis meses de cultivo, amostras compostas de folhas foram coletadas e submetidas ao processo de hidrodestilação, por meio do aparelho do tipo Clevenger, para quantificação do óleo essencial. Em solo com acúmulo de matéria orgânica as doses e compostos avaliados não influenciaram o teor de óleo essencial de Ocimum selloi Beth.Disponível também em: Cadernos de Agroecologia, V. 5, n.1, 2010

PIP3-Independent Activation of TorC2 and PKB at the Cell's Leading Edge Mediates Chemotaxis

SummaryBackgroundStudies show that high phosphotidylinositol 3,4,5-trisphosphate (PIP3) promotes cytoskeletal rearrangements and alters cell motility and chemotaxis, possibly through activation of protein kinase Bs (PKBs). However, chemotaxis can still occur in the absence of PIP3, and the identities of the PIP3-independent pathways remain unknown.ResultsHere, we outline a PIP3-independent pathway linking temporal and spatial activation of PKBs by Tor complex 2 (TorC2) to the chemotactic response. Within seconds of stimulating Dictyostelium cells with chemoattractant, two PKB homologs, PKBA and PKBR1, mediate transient phosphorylation of at least eight proteins, including Talin, PI4P 5-kinase, two Ras GEFs, and a RhoGap. Surprisingly, all of the substrates are phosphorylated with normal kinetics in cells lacking PI 3-kinase activity. Cells deficient in TorC2 or PKB activity show reduced phosphorylation of the endogenous substrates and are impaired in chemotaxis. The PKBs are activated through phosphorylation of their hydrophobic motifs via TorC2 and subsequent phosphorylation of their activation loops. These chemoattractant-inducible events are restricted to the cell's leading edge even in the absence of PIP3. Activation of TorC2 depends on heterotrimeric G protein function and intermediate G proteins, including Ras GTPases.ConclusionsThe data lead to a model where cytosolic TorC2, encountering locally activated small G protein(s) at the leading edge of the cell, becomes activated and phosphorylates PKBs. These in turn phosphorylate a series of signaling and cytoskeletal proteins, thereby regulating directed migration

Measurement of Length and Position with Frequency Combs

We show two different absolute distance measurement methods with micrometer accuracy based on frequency combs, and we discuss possible applications. Using a mode- locked laser and MEMS-based tracking optics, we measure the 3D position of a retroreflector within 10 ms and with a 24 μm volumetric accuracy. We also investigate modulator-based combs and show that they enable highly sensitive surface topography measurements with microsecond acquisition times and micrometer precision. Moreover, the potential for photonic integration of frequency comb sources is explored

The measurement of volatile organic compounds in faeces of piglets as a tool to assess gastrointestinal functionality

There is an increasing interest in developing innovative means to monitor animal health through precision farming. As part of this drive, we have targeted digestive health and in particular the microbiota. In this study, we investigated the effect of different dietary interventions in piglets, feeding these piglets with one of two different feeds (high protein and low protein). We then evaluated its effects by measuring the volatile organic compounds (VOC) that emanated from these faecal samples using various forms of Ion Mobility Spectrometry. Piglets were monitored for 19 days, with faecal samples collected on days 6, 12 and 19, providing a total of 69 samples. The statistical analysis attempted to separate the samples using either dietary intervention or faecal score. First, the faecal score was investigated using a value based on a visual faecal scoring system, from 0 to 3, with 0 being normal and 3 having diarrhoea. Then the VOCs were analysed in regard to dietary intervention (high vs low protein). Results indicated that our approach was able to separate the dietary intervention (area under the curve (AUC) 0.81) using VOC data. Furthermore, we were able to separate samples based on faecal score (AUC between 0.71 and 1, with six different comparisons undertaken). We believe that faeces volatiles hold potential as a future means to monitor animal health

The role of hippocampal NMDA receptors in long-term emotional responses following muscarinic receptor activation

Extensive evidence indicates the influence of the cholinergic system on emotional processing. Previous findings provided new insights into the underlying mechanisms of long-term anxiety, showing that rats injected with a single systemic dose of pilocarpine--a muscarinic receptor (mAChR) agonist--displayed persistent anxiogenic-like responses when evaluated in different behavioral tests and time-points (24 h up to 3 months later). Herein, we investigated whether the pilocarpine-induced long-term anxiogenesis modulates the HPA axis function and the putative involvement of NMDA receptors (NMDARs) following mAChRs activation. Accordingly, adult male Wistar rats presented anxiogenic-like behavior in the elevated plus-maze (EPM) after 24 h or 1 month of pilocarpine injection (150 mg/kg, i.p.). In these animals, mAChR activation disrupted HPA axis function inducing a long-term increase of corticosterone release associated with a reduced expression of hippocampal GRs, as well as consistently decreased NMDAR subunits expression. Furthermore, in another group of rats injected with memantine--an NMDARs antagonist (4 mg/kg, i.p.)--prior to pilocarpine, we found inhibition of anxiogenic-like behaviors in the EPM but no further alterations in the pilocarpine-induced NMDARs downregulation. Our data provide evidence that behavioral anxiogenesis induced by mAChR activation effectively yields short- and long-term alterations in hippocampal NMDARs expression associated with impairment of hippocampal inhibitory regulation of HPA axis activity. This is a novel mechanism associated with anxiety-like responses in rats, which comprise a putative target to future translational studies