Longitudinal evolution of the immune suppressive glioma microenvironment in different synchronous lesions during treatment

The role of immune suppression in glioma progression has been clearly established.1 We and others have recently demonstrated that myeloid cells play a major role in the tumor microenvironment of glioblastoma (GBM) patients,2,3 and that not only bone marrow-derived macrophages (BMDMs) have a higher intrinsic immune suppressive ability compared to resident microglial cells (MG), but also that this ability greatly increases going from the periphery to the tumor core.3 In lower grade gliomas (grades II and III), a much lower amount of BMDM is present, devoid of immune suppressive ability.3 We present here a longitudinal analysis of the immune infiltrate in a patient with a synchronous occurrence of GBM in the left temporal lobe, and a low-grade glioma (LGG) in the right frontal lobe, with discordant isocitrate dehydrogenase (IDH)-mutational status,4 followed by two GBM relapse

K+ accumulation and clearance in the calyx synaptic cleft of type I mouse vestibular hair cells

Vestibular organs of Amniotes contain two types of sensory cells, named Type I and Type II hair cells. While Type II hair cells are contacted by several small bouton nerve terminals, Type I hair cells receive a giant terminal, called a calyx, which encloses their basolateral membrane almost completely. Both hair cell types release glutamate, which depolarizes the afferent terminal by binding to AMPA post-synaptic receptors. However, there is evidence that non-vesicular signal transmission also occurs at the Type I hair cell-calyx synapse, possibly involving direct depolarization of the calyx by K+ exiting the hair cell. To better investigate this aspect, we performed whole-cell patch-clamp recordings from mouse Type I hair cells or their associated calyx. We found that [K+] in the calyceal synaptic cleft is elevated at rest relative to the interstitial (extracellular) solution and can increase or decrease during hair cell depolarization or repolarization, respectively. The change in [K+] was primarily driven by GK,L, the low-voltage-activated, non-inactivating K+ conductance specifically expressed by Type I hair cells. Simple diffusion of K+ between the cleft and the extracellular compartment appeared substantially restricted by the calyx inner membrane, with the ion channels and active transporters playing a crucial role in regulating intercellular [K+]. Calyx recordings were consistent with K+ leaving the synaptic cleft through postsynaptic voltage-gated K+ channels involving KV1 and KV7 subunits. The above scenario is consistent with direct depolarization and hyperpolarization of the calyx membrane potential by intercellular K+

Optimizing the vertebrate vestibular semicircular canal: could we balance any better?

The fluid-filled semicircular canals (SCCs) of the vestibular system are used by all vertebrates to sense angular rotation. Despite masses spanning seven decades, all mammalian SCCs are nearly the same size. We propose that the SCC represents a sensory organ that evolution has `optimally designed'. Four geometric parameters are used to characterize the SCC, and `building materials' of given physical properties are assumed. Identifying physical and physiological constraints on SCC operation, we find that the most sensitive SCC has dimensions consistent with available data.Comment: 4 pages, 3 figure

Row spacing and seed physiological quality of Crotalaria species.

Crotalaria crops have important uses, such as in green manuring, nematode control, biological nitrogen fixation and sugarcane reform in Savanna areas. Due to its strategic importance, knowledge about crotalaria seed production technology is a relevant factor to ensure the availability of high physiological quality seeds

Targeting of immunosuppressive myeloid cells from glioblastoma patients by modulation of size and surface charge of lipid nanocapsules

Background: Myeloid derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs) are two of the major players involved in the inhibition of anti-tumor immune response in cancer patients, leading to poor prognosis. Selective targeting of myeloid cells has therefore become an attractive therapeutic strategy to relieve immunosuppression and, in this frame, we previously demonstrated that lipid nanocapsules (LNCs) loaded with lauroyl-modified gemcitabine efficiently target monocytic MDSCs in melanoma patients. In this study, we investigated the impact of the physico-chemical characteristics of LNCs, namely size and surface potential, towards immunosuppressive cell targeting. We exploited myeloid cells isolated from glioblastoma patients, which play a relevant role in the immunosuppression, to demonstrate that tailored nanosystems can target not only tumor cells but also tumor-promoting cells, thus constituting an efficient system that could be used to inhibit their function. Results: The incorporation of different LNC formulations with a size of 100 nm, carrying overall positive, neutral or negative charge, was evaluated on leukocytes and tumor-infiltrating cells freshly isolated from glioblastoma patients. We observed that the maximum LNC uptake was obtained in monocytes with neutral 100 nm LNCs, while positively charged 100 nm LNCs were more effective on macrophages and tumor cells, maintaining at low level the incorporation by T cells. The mechanism of uptake was elucidated, demonstrating that LNCs are incorporated mainly by caveolae-mediated endocytosis. Conclusions: We demonstrated that LNCs can be directed towards immunosuppressive cells by simply modulating their size and charge thus providing a novel approach to exploit nanosystems for anticancer treatment in the frame of immunotherapy.[Figure not available: see fulltext.

Alterações da disponibilidade de nutrientes em um Latossolo Vermelho-Amarelo degradado e outro não-degradado fertilizados com biossólido e florestados com Eucalyptus grandis.

O presente trabalho teve como objetivos avaliar os efeitos da aplicação de biossólido sobre as modificações de fertilidade do solo e o crescimento de povoamentos de Eucalyptus grandis plantados em um Latossolo Vermelho-Amarelo Distrófico não-degradado e outro degradado

Myeloid Diagnostic and Prognostic Markers of Immune Suppression in the Blood of Glioma Patients.

Although gliomas are confined to the central nervous system, their negative influence over the immune system extends to peripheral circulation. The immune suppression exerted by myeloid cells can affect both response to therapy and disease outcome. We analyzed the expansion of several myeloid parameters in the blood of low- and high-grade gliomas and assessed their relevance as biomarkers of disease and clinical outcome. Methods: Peripheral blood was obtained from 134 low- and high-grade glioma patients. CD14+, CD14+/p-STAT3+, CD14+/PD-L1+, CD15+ cells and four myeloid-derived suppressor cell (MDSC) subsets, were evaluated by flow cytometry. Arginase-1 (ARG1) quantity and activity was determined in the plasma. Multivariable logistic regression model was used to obtain a diagnostic score to discriminate glioma patients from healthy controls and between each glioma grade. A glioblastoma prognostic model was determined by multiple Cox regression using clinical and myeloid parameters. Results: Changes in myeloid parameters associated with immune suppression allowed to define a diagnostic score calculating the risk of being a glioma patient. The same parameters, together with age, permit to calculate the risk score in differentiating each glioma grade. A prognostic model for glioblastoma patients stemmed out from a Cox multiple analysis, highlighting the role of MDSC, p-STAT3, and ARG1 activity together with clinical parameters in predicting patient's outcome. Conclusions: This work emphasizes the role of systemic immune suppression carried out by myeloid cells in gliomas. The identification of biomarkers associated with immune landscape, diagnosis, and outcome of glioblastoma patients lays the ground for their clinical use

Current response in CaV1.3–/– mouse vestibular and cochlear hair cells

Signal transmission by sensory auditory and vestibular hair cells relies upon Ca2+-dependent exocytosis of glutamate. The Ca2+ current in mammalian inner ear hair cells is predominantly carried through CaV1.3 voltage-gated Ca2+ channels. Despite this, CaV1.3 deficient mice (CaV1.3–/–) are deaf but do not show any obvious vestibular phenotype. Here, we compared the Ca2+ current (ICa) in auditory and vestibular hair cells from wild-type and CaV1.3–/– mice, to assess whether differences in the size of the residual ICa could explain, at least in part, the two phenotypes. Using 5 mM extracellular Ca2+ and near-body temperature conditions, we investigated the cochlear primary sensory receptors inner hair cells (IHCs) and both type I and type II hair cells of the semicircular canals. We found that the residual ICa in both auditory and vestibular hair cells from CaV1.3–/– mice was less than 20% (12–19%, depending on the hair cell type and age investigated) compared to controls, indicating a comparable expression of CaV1.3 Ca2+ channels in both sensory organs. We also showed that, different from IHCs, type I and type II hair cells from CaV1.3–/– mice were able to acquire the adult-like K+ current profile in their basolateral membrane. Intercellular K+ accumulation was still present in CaV1.3–/– mice during IK,L activation, suggesting that the K+-based, non-exocytotic, afferent transmission is still functional in these mice. This non-vesicular mechanism might contribute to the apparent normal vestibular functions in CaV1.3–/– mice

Techonolgy of Qualea grandiflora Mart. (Vochysiaceae) seeds

Qualea grandiflora Mart. (Vochysiaceae), commonly known as "pau-terra", is an arborous species native to the Brazilian savannah which possess commercial interests, as it can be used either as an ornamental or as a medicinal plant. "Pau-terra" can also be used in the heterogeneous reforestation of areas which are destined for restoration of permanent preservation degraded areas. Propagation studies with this species are scarce, being necessary then further clarification regarding the factors that influences the germination process. In this context, the objective of this work was to evaluate the influence of different temperatures, substrates and light conditions on seed germination. We selected light brown seeds which were subjected to different interactions between temperatures (15-25, 20-30, 25 and 30°C), substrate (paper, sand and vermiculite) and light (light and dark). All seeds were later dry-incubated at 32°C for 3, 6 and 12 hours. After treatments, seeds were kept in BOD at 58% RH and the following parameters were calculated: germination (%G) and germination speed index (GSI); the formation of normal and abnormal seedlings and the number dead seeds. Interaction was observed for all variables. In the optimum temperature range, the seeds behaved as photoblastic neutral or indifferent. Under alternating temperatures, darkness enhanced the germination, especially when combined with the lower temperatures. We noted that the sowing in sand, at 25°C, allowed the maintenance of suitable combinations of germination and seedling development. With respect to desiccation tolerance, "pau-terra" seeds presented an orthodox behavior, with a linear increase of the vigor as function of drying