In vitro culture from mature seeds of Passiflora species

The genus Passiflora comprises hundred species, mainly native of the South American tropics and rainforests, which are grouped into 21 subgenera. Some species are widely studied for their economic importance and are chiefly cultivated for production of fruit juice. To obtain a continuous source of material for a screening of secondary metabolites, zygotic embryo culture was attempted for 62 Passiflora species, starting from seeds mainly collected in the wild. Twenty nine of these species produced calli, which had very different growth rates. Plants were successfully regenerated from calli of 13 different species. For 25 of the responsive species this is the first report of in vitro culture

Regeneração e cultura in vitro de espécies de Passiflora

O gênero Passiflora compõe centenas de espécies, a maioria de origem dos trópicos e das florestas da América do Sul, as quais são agrupadas em 21 subgêneros. Algumas espécies foram intensamente estudadas por sua importância econômica e são cultivadas principalmente para a produção de suco de fruta. Cultura de 29 espécies de Passiflora foram obtidos a partir de embriões zigóticos e de culturas de endosperma. Foram obtidos diferentes tipos de calos de crescimento, de tal forma que plantas foram regeneradas a partir de calos de 13 espécies diferentes. Não haviam sido ainda relatadas culturas in vitro para 25 das espécies trabalhadas.The genus Passiflora comprises hundred species, mainly native of the South American tropics and rainforests, which are grouped into 21 subgenera. Some species are widely studied for their economic importance and are chiefly cultivated for production of fruit juice. To obtain a continuous source of material for a screening of secondary metabolites, zygotic embryo culture was attempted for 62 Passiflora species, starting from seeds mainly collected in the wild. Twenty nine of these species produced calli, which had very different growth rates. Plants were successfully regenerated from calli of 13 different species. For 25 of the responsive species this is the first report of in vitro culture

Assessment of immunostimulatory responses to the antimiR-22 oligonucleotide compound RES-010 in human peripheral blood mononuclear cells

microRNA-22 (miR-22) is a key regulator of lipid and energy homeostasis and represents a promising therapeutic target for NAFLD and obesity. We have previously identified a locked nucleic acid (LNA)-modified antisense oligonucleotide compound complementary to miR-22, designated as RES-010 that mediated robust inhibition of miR-22 function in cultured cells and in vivo. In this study we investigated the immune potential of RES-010 in human peripheral blood mononuclear cells (PBMCs). We treated fresh human peripheral blood mononuclear cells isolated from six healthy volunteers with different concentrations of the RES-010 compound and assessed its proinflammatory effects by quantifying IL-1β, IL-6, IFN-γ, TNF-α, IFN-α2a, IFN-β, IL-10, and IL-17A in the supernatants collected 24 h of treatment with RES-010. The T-cell activation markers, CD69, HLA-DR, and CD25 were evaluated by flow cytometry after 24 and 144 h of treatment, respectively, whereas cell viability was assessed after 24 h of treatment with RES-010. Our results show that RES-010 compound does not induce any significant immunostimulatory responses in human PBMCs in vitro compared to controls, implying that the proinflammatory potential of RES-010 is low.</p

Can early-onset acquired demyelinating syndrome (ADS) hide pediatric Behcet's disease? A case report

Behcet's disease (BD) is a rare vasculitis characterized by multisystemic inflammation. Central nervous system (CNS) involvement is rare and heterogeneous, particularly in the pediatric population. A diagnosis of neuro-Behcet could be highly challenging, especially if neurological manifestations precede other systemic features; however, its timely definition is crucial to prevent long-term sequelae. In this study, we describe the case of a girl who, at 13 months of age, presented with a first episode of encephalopathy compatible with acute disseminated encephalomyelitis, followed, after 6 months, by a neurological relapse characterized by ophthalmoparesis and gait ataxia, in association with new inflammatory lesions in the brain and spinal cord, suggesting a neuromyelitis optica spectrum disorder. The neurological manifestations were successfully treated with high-dose steroids and intravenous immunoglobulins. In the following months, the patient developed a multisystemic involvement suggestive of Behcet's disease, characterized by polyarthritis and uveitis, associated with HLA-B51 positivity. The challenge presented by this unique case required a multidisciplinary approach involving pediatric neurologists, neuro-radiologists, and pediatric rheumatologists, with all of these specialists creating awareness about early-onset acquired demyelinating syndromes (ADSs). Given the rarity of this presentation, we performed a review of the literature focusing on neurological manifestations in BD and differential diagnosis of patients with early-onset ADS

Serotonergic system and its interaction with neuroinflammation

Serotonin (5-HT) is a neurotransmitter that is mainly expressed in brain where serves a wide array of physiological and behavioural functions. Literature described that some mediators of inflammation (i.e. cytokines) have been implicated in the modulation of monoaminergic response and this may be associated with pathophysiology of depression and in the responsiveness of antidepressant treatment in both humans and animals (Capuron and Miller, 2011). A hypothesis suggests that cytokines may affect the serotonergic system through p38 MAP kinase dependent mechanisms particularly at the serotonin transporters (Zhu et al., 2006) and 5HT7 receptors (Lieb et al., 2005; Mahe et al., 2005). The aim of this study was to show the interaction of Interleukin 1β (IL-1β) or p38 MAP kinase on serotonin transporter (SERT) and 5HT7 receptors in cells lines and native tissue, highlighting the biochemical mechanism of this system. The IL-1β and p38 MAP kinase activator, anisomycin, did not show any effect on 5-HT uptake and p38 MAPK activation in rodent native brain tissue, in human platelets and in cell lines in contrast to literature reports (Zhu et al.,2010). A different method was then used in which a release of cytokines was induced directly in the rat brain through an i.c.v. LPS treatment. Although proinflammatory cytokines involved in the change of animal mood, such as IL-1ß and TNFα, showed a significant increase in cortex and striatum, a modulation of SERT activity in term of Km and Vmax was not detected, confirming again that no interaction between cytokines, p38 MAP kinase and SERT function in vitro nor in vivo was evident. In contrast, this study revealed a positive interaction between 5HT7 receptors and p38 MAP kinase in glia cells. However, this pathway was not present in cortical neurons where 5HT7 receptors did not activate p38 MAP kinase but instead increased the AMPAR subunit, GluR1 and CREB phosphorylation. The effect on GluR1 was reversed by the specific 5HT7 antagonist, SB258719, and the PKA inhibitor, H89, confirming the specificity of response for 5HT7 receptors and the involvement of PKA in the mediation of GluR1 phosphorylation. In conclusion, this study displayed a lack of interaction between IL-1β and p38 MAP kinase on rat SERT while highlighting the effect of 5HT7 receptors on p38 MAP kinase, with different functions between glial and neuronal cells. Noteworthy, this is the first report that showed a positive interaction between 5HT7 receptors and AMPA which stimulates new investigation into the role of 5HT7 receptors in neuronal plasticity

p38 MAP kinase activation does not stimulate serotonin transport in rat brain:Implications for sickness behaviour mechanisms

none4siAims Several studies suggested an association between dysregulation of immune mediators and behavioural, neuroendocrine and neurochemical features of depression. Available data showed that cytokines affect the serotonin transporter (SERT) activity through p38 MAP kinase (MAPK)-dependent mechanisms in some cell lines and mice neurons (Zhu et al., Neuropsychopharmacology, 2006; 31:2121–31). The aim of this study was to investigate the interaction of Interleukin-1β (IL-1β) or p38 MAPK with SERT activity in rat brain and cell lines. Main methods Synaptosomes or cells were treated with IL-1β or the p38 MAPK activator anisomycin at different concentrations and end-points and the modulation of SERT activity as Km and Vmax was evaluated. Key findings Treatments with IL-1β or anisomycin did not affect serotonin uptake and p38 MAPK activation in rat synaptosomes, in contrast to reports in mice (Zhu et al., Neuropsychopharmacology, 2010; 35:2510–20). The same treatments activated p38 MAPK phosphorylation in HeLa cells used as positive controls. Similarly, no changes after anisomycin treatment could be detected in [3H]serotonin uptake rate in LLC-PK cells expressing human SERT, although phosphorylated p38 MAPK levels augmented significantly. Direct cytokine release in brain was induced by intracerebroventricular administration of bacterial lipopolysaccaride. Although pro-inflammatory cytokines, such as IL-1ß, IL6, and Tumor Necrosis Factor α, showed significant increases in brain cortex, modulation of SERT activity in term of Km and Vmax was not detected. Significance These results imply that the stimulation of serotonin uptake by cytokines may not be a unique and fundamental mechanism in the pathology of depression induced by altered immune response.mixedFilippo Andreetta; Nicholas M. Barnes; Paul B. Wren; Lucia CarboniFilippo Andreetta; Nicholas M. Barnes; Paul B. Wren; Lucia Carbon