A tree nymph of the Brazilian Atlantic Forest: Dryades (Galipeinae, Rutaceae), a new neotropical genus segregated from Conchocarpus.

Subtribe Galipeinae (tribe Galipeeae) is the most diverse group of Rutaceae (the orange family) in the Neotropics, with 27 genera and ca. 130 species. The largest genus in the subtribe is Conchocarpus, with ca. 50 species, distributed from Central America to southern Brazil, and is particularly diverse in the Brazilian Atlantic Forest. The circumscription of the genus was recently changed to accommodate the species of Almeidea. However, even with this inclusion, Conchocarpus did not appear as monophyletic because the position of C. concinnus, which appeared in a clade with the other genera of Galipeinae rather than in the clade with the other species of Conchocarpus. The objective of the present study is to investigate the phylogenetic position of four other species of Conchocarpus (hereafter called "C. gauchaudianus group") that share morphological traits and geographical distribution with C. concinnus suggesting a close phylogenetic affinity. Phylogenetic analyses were based on morphological and molecular data from nuclear regions ITS-1 and ITS-2 as well as plastid regions trnL-trnF and rps-16, and were conducted with parsimony and Bayesian inference as optimization criteria. Results showed Conchocarpus as polyphyletic with its species divided in two clades, one, herein called "the Conchocarpus sensu stricto group," includes the type species C. macrophyllus, and the other "the Conchocarpus gaudichaudianus group" includes C. concinnus. The latter group is here recognized as a new genus, Dryades, the name given by Carl Friederich von Martius (1794-1868) to the Domain of the Atlantic Forest in Brazil, inspired by the tree nymphs in Greek mythology. Floral structure and leaf morphology provided further support to the findings of phylogenetic analysis. A description of the new genus, new combinations, a key to the species of the new genus, discussions of the affinities of the species are also provided, as well as data on the conservation status of the species of Dryades. Additionally, new data on floral structure of C. heterophyllus, C. macrophyllus and C. minutiflorus (all from the Conchocarpus sensu stricto group) are provided.

Host dysbiosis negatively impacts IL-9-producing T-cell differentiation and antitumour immunity.

BACKGROUND: Host-microbiota interactions shape T-cell differentiation and promote tumour immunity. Although IL-9-producing T cells have been described as potent antitumour effectors, their role in microbiota-mediated tumour control remains unclear. METHODS: We analysed the impact of the intestinal microbiota on the differentiation of colonic lamina propria IL-9-producing T cells in germ-free and dysbiotic mice. Systemic effects of the intestinal microbiota on IL-9-producing T cells and the antitumour role of IL-9 were analysed in a model of melanoma-challenged dysbiotic mice. RESULTS: We show that germ-free mice have lower frequency of colonic lamina propria IL-9-producing T cells when compared with conventional mice, and that intestinal microbiota reconstitution restores cell frequencies. Long-term antibiotic treatment promotes host dysbiosis, diminishes intestinal IL-4 and TGF-ß gene expression, decreases the frequency of colonic lamina propria IL-9-producing T cells, increases the susceptibility to tumour development and reduces the frequency of IL-9-producing T cells in the tumour microenvironment. Faecal transplant restores intestinal microbiota diversity, and the frequency of IL-9-producing T cells in the lungs of dysbiotic animals, restraining tumour burden. Finally, recombinant IL-9 injection enhances tumour control in dysbiotic mice. CONCLUSIONS: Host-microbiota interactions are required for adequate differentiation and antitumour function of IL-9-producing T cells.

Beneficial Propionibacteria within a Probiotic Emmental Cheese: Impact on Dextran Sodium Sulphate-Induced Colitis in Mice.

BACKGROUNDS AND AIMS: Inflammatory Bowel Diseases (IBD), including Ulcerative Colitis (UC), coincide with alterations in the gut microbiota. Consumption of immunomodulatory strains of probiotic bacteria may induce or prolong remission in UC patients. Fermented foods, including cheeses, constitute major vectors for bacteria consumption. New evidences revealed anti-inflammatory effects in selected strains of Propionibacterium freudenreichii. We thus hypothesized that consumption of a functional cheese, fermented by such a strain, may exert a positive effect on IBD. METHODS: We investigated the impact of cheese fermented by P. freudenreichii on gut inflammation. We developed an experimental single-strain cheese solely fermented by a selected immunomodulatory strain of P. freudenreichii, CIRM-BIA 129. We moreover produced, in industrial conditions, an Emmental cheese using the same strain, in combination with Lactobacillus delbrueckii CNRZ327 and Streptococcus thermophilus LMD-9, as starters. Consumption of both cheeses was investigated with respect to prevention of Dextran Sodium Sulphate (DSS)-induced colitis in mice. RESULTS: Consumption of the single-strain experimental cheese, or of the industrial Emmental, both fermented by P. freudenreichii CIRM-BIA 129, reduced severity of subsequent DSS-induced colitis, weight loss, disease activity index and histological score. Both treatments, in a preventive way, reduced small bowel Immunoglobulin A (IgA) secretion, restored occludin gene expression and prevented induction of Tumor Necrosis Factor α (TNFα), Interferon γ (IFNγ) and Interleukin-17 (IL-17). CONCLUSIONS: A combination of immunomodulatory strains of starter bacteria can be used to manufacture an anti-inflammatory cheese, as revealed in an animal model of colitis. This opens new perspectives for personalised nutrition in the context of IBD.

Early life history of the Caribbean coral Orbicella faveolata (Scleractinia: Merulinidae) / Historia de vida temprana del coral caribeño Orbicella faveolata (Scleractinia: Merulinidae)

Introduction: Rehabilitation of hermatypic coral species that have declined in the Caribbean in recent decades is a priority. Production of sexual recruits is considered the best restoration method to aid affected populations. Objective: To gain knowledge of early life stages of Orbicella faveolata and to enhance production of new sexual recruits. Methods: Gamete bundles from the coral species O. faveolata were collected over two years (2018 and 2019) from Los Corales del Rosario y de San Bernardo Natural National Park, Cartagena, Colombia. Assisted fertilization, larval rearing, settlement (onto crustose coralline algae, CCA) and post settlement survival in laboratory conditions were monitored. Results: Embryonic and larval development were documented over 55 hours after the first cleavage, when larvae were fully developed and started pre-settlement behavior. Settlement began 7 days after first cleavage and after 37 days polyps had acquired zooxanthellae. Larval settlement was higher on Lythophyllum congestum and Titanoderma prototypum than in response to Porolithon pachydermum, Neogoniolithon sp., Hydrolithon sp., and Lythophyllum sp. Larvae did not settle on dead coral or on the negative control (sterilized seawater). After the first week post settlement survival was 59 % amongst O. faveolata recruits. During the second week, survival dropped to 42 %, and was further reduced to 0 % at the end of the third week. Conclusions: O. faveolata larvae require cues from certain CCA species to settle, they do not settle in absence of CCA. Increased larvae availability is possible through assisted fertilization in the laboratory, however, due to the high mortality in early post-settlement phases, additional research needs to be conducted in order to scale up larvae production and improve understanding of the cues that enhance settlement and the factors which cause post-settlement mortality.

Introducción: La rehabilitación de las especies de corales hermatípicos del Caribe que han disminuido en las últimas décadas es una prioridad. La producción de reclutas sexuales se considera el mejor método de restauración para ayudar a las poblaciones afectadas. Objetivo: Obtener conocimiento de las primeras etapas de la vida de O. faveolata y mejorar la producción de nuevos reclutas sexuales. Métodos: Por dos años (2018 y 2019), seis días después de luna llena en septiembre, se recolectaron paquetes gaméticos en arrecifes del Parque Nacional Natural Los Corales del Rosario y de San Bernardo, Cartagena, Colombia. Se siguió la fertilización asistida, la cría de larvas, el asentamiento y la supervivencia posterior al asentamiento en algas coralinas costrosas (ACC) en condiciones de laboratorio. Resultados: El desarrollo de embriones y larvas se documenta a lo largo de 55 h después del primer clivaje, cuando la larva está desarrollada completamente y comenzó el comportamiento previo al asentamiento. El asentamiento comienza 7 días después del primer clivaje y 37 días después, la mayoría de los pólipos presentan zooxantelas. El asentamiento larval fue más alto en Lythophyllum congestum y Titanoderma prototypum que en respuesta a Porolithon pachydermum, Neogoniolithon sp., Hydrolithon sp., y Lythophyllum sp. No hubo asentamiento sobre coral muerto ni en el control negativo (agua de mar esterilizada). La supervivencia bajó de un 59 % en la primera semana después del asentamiento, a 42 % durante la segunda semana y 0 % para el final de la tercera semana. Conclusiones: Las larvas de O. faveolata requieren señales de ciertas especies de ACC para asentarse, ellas no se asientan en ausencia de ACC. La disponibilidad de larvas es posible mediante la fertilización asistida en laboratorio. Sin embargo, debido a la alta mortalidad en las primeras fases posteriores al asentamiento, queda mucho por hacer para aumentar la producción de larvas y mejorar nuestro conocimiento y comprensión de las señales que mejoran el asentamiento y las que previenen o inhiben la supervivencia del recluta.

Probiotic Propionibacterium freudenreichii requires SlpB protein to mitigate mucositis induced by chemotherapy.

Propionibacterium freudenreichii CIRM-BIA 129 (P. freudenreichii wild type, WT) is a probiotic bacterium, which exerts immunomodulatory effects. This strain possesses extractable surface proteins, including SlpB, which are involved in anti-inflammatory effect and in adhesion to epithelial cells. We decided to investigate the impact of slpB gene mutation on immunomodulation in vitro and in vivo. In an in vitro assay, P. freudenreichii WT reduced expression of IL-8 (p<0.0001) and TNF-α (p<0.0001) cytokines in LPS-stimulated HT-29 cells. P. freudenreichii ΔslpB, lacking the SlpB protein, failed to do so. Subsequently, both strains were investigated in vivo in a 5-FU-induced mucositis mice model. Mucositis is a common side effect of cytotoxic chemotherapy with 5-FU, characterized by mucosal injury, inflammation, diarrhea, and weight loss. The WT strain prevented weight loss, reduced inflammation and consequently histopathological scores. Furthermore, it regulated key markers, including Claudin-1 (cld1, p<0.0005) and IL-17a (Il17a, p<0.0001) genes, as well as IL-12 (p<0.0001) and IL-1ß (p<0.0429) cytokines levels. Mutant strain displayed opposite regulatory effect on cld1 expression and on IL-12 levels. This work emphasizes the importance of SlpB in P. freudenreichii ability to reduce mucositis inflammation. It opens perspectives for the development of probiotic products to decrease side effects of chemotherapy using GRAS bacteria with immunomodulatory surface protein properties.

Whey Protein Isolate-Supplemented Beverage, Fermented by Lactobacillus casei BL23 and Propionibacterium freudenreichii 138, in the Prevention of Mucositis in Mice.

Mucositis is a clinically important gastrointestinal inflammatory infirmity, generated by antineoplastic drugs cytotoxic effects. The inflammatory process caused by this disease frequently leads to derangements in the alimentary tract and great malaise for the patient. Novel strategies are necessary for its prevention or treatment, as currently available treatments of mucositis have several limitations in relieving its symptoms. In this context, several research groups have investigated the use of probiotic bacteria, and in particular dairy bacterial strains. Compelling evidences reveal that milk fermented by certain probiotic bacteria has the capacity to ameliorate intestinal inflammatory disorders. In addition, innovative probiotic delivery strategies, based on probiotics incorporation into protective matrices, such as whey proteins, were able to increase the therapeutic effect of probiotic strains by providing extra protection for bacteria against environmental stresses. Therefore, in this study, we evaluated the role of the whey protein isolate (WPI), when added to skim milk fermented by Lactobacillus casei BL23 (L. casei BL23) or by Propionibacterium freudenreichii CIRM-BIA138 (P. freudenreichii 138), as a protective matrix against in vitro stress challenges. In addition, we investigated the therapeutic effect of these fermented beverages in a murine model of mucositis induced by 5-Fluorouracil (5-FU). Our results demonstrated that milk supplementation with 30% (w/v) of WPI increases the survival rate of both strains when challenged with acid, bile salts, high temperature and cold storage stresses, compared to fermented skim milk without the addition of WPI. Moreover, treatment with the probiotic beverages prevented weight loss and intestinal damages in mice receiving 5-FU. We conclude that the presence of WPI maximizes the anti-inflammatory effects of L. casei BL23, but not for P. freudenreichii 138, suggesting that whey protein enhancement of probiotic activity might be strain-dependent.

Secretion of biologically active pancreatitis-associated protein I (PAP) by genetically modified dairy Lactococcus lactis NZ9000 in the prevention of intestinal mucositis.

BACKGROUND: Mucositis is one of the most relevant gastrointestinal inflammatory conditions in humans, generated by the use of chemotherapy drugs, such as 5-fluoracil (5-FU). 5-FU-induced mucositis affects 80% of patients undergoing oncological treatment causing mucosal gut dysfunctions and great discomfort. As current therapy drugs presents limitations in alleviating mucositis symptoms, alternative strategies are being pursued. Recent studies have shown that the antimicrobial pancreatitis-associated protein (PAP) has a protective role in intestinal inflammatory processes. Indeed, it was demonstrated that a recombinant strain of Lactococcus lactis expressing human PAP (LL-PAP) could prevent and improve murine DNBS-induced colitis, an inflammatory bowel disease (IBD) that causes severe inflammation of the colon. Hence, in this study we sought to evaluate the protective effects of LL-PAP on 5-FU-induced experimental mucositis in BALB/c mice as a novel approach to treat the disease. RESULTS: Our results show that non-recombinant L. lactis NZ9000 have antagonistic activity, in vitro, against the enteroinvasive gastrointestinal pathogen L. monocytogenes and confirmed PAP inhibitory effect against Opportunistic E. faecalis. Moreover, L. lactis was able to prevent histological damage, reduce neutrophil and eosinophil infiltration and secretory Immunoglobulin-A in mice injected with 5-FU. Recombinant lactococci carrying antimicrobial PAP did not improve those markers of inflammation, although its expression was associated with villous architecture preservation and increased secretory granules density inside Paneth cells in response to 5-FU inflammation. CONCLUSIONS: We have demonstrated for the first time that L. lactis NZ9000 by itself, is able to prevent 5-FU-induced intestinal inflammation in BALB/c mice. Moreover, PAP delivered by recombinant L. lactis strain showed additional protective effects in mice epithelium, revealing to be a promising strategy to treat intestinal mucositis.

Vitamin A supplementation leads to increases in regulatory CD4+Foxp3+LAP+ T cells in mice.

Dietary compounds, including micronutrients such as vitamin A and its metabolite retinoic acid, directly influence the development and function of the immune system. In this study, we show that either dietary deficiency of or supplementation with vitamin A had immunologic effects in mice that were fed these diets during their development (for 8 wk during the postweaning period). Deficient mice presented higher levels of interferon-γ, interleukin (IL)-6, transforming growth factor-ß, IL-17, and IL-10 in the gut-associated lymphoid tissues and draining lymph nodes, indicating a proinflammatory shift in the gut mucosa. Serum immunoglobulin G levels also were elevated in these mice. Conversely, supplemented mice showed higher frequencies of CD4+Foxp3+LAP+ regulatory T cells in gut lymphoid tissues and spleen, suggesting that vitamin A supplementation in the diet may be beneficial in pathologic situations such as inflammatory bowel diseases.

Hypercholesterolemia and neurodegeneration. Comparison of hippocampal phenotypes in LDLr knockout and APPswe/PS1dE9 mice.

Previous studies suggest that Alzheimer's disease (AD) neurobiology could not be explained solely by an increase in ß-amyloid levels. Recently, it has been proposed that alterations in brain cholesterol metabolism may contribute to the pathogenesis of AD. In the present work, we focus on early changes in the hippocampal phenotypes of two mouse models in which cognitive impairments were previously described: a) the hypercholesterolemic LDL receptor knockout (LDLr -/-) and b) the APPswe/PS1dE9 (APP/PS1) transgenic model of familial AD. Our initial analysis, subsequent validation and additional experiments at the mRNA and protein levels demonstrate some parallels between the hippocampal phenotypes of these 2 mouse models, however our data suggest that the molecular mechanisms leading to cognitive decline are distinct in LDLr -/- and APP/PS1 animals. Genes related to cytokine signaling were significantly down-regulated in LDLr -/- mice when compared to both the wild-type and APP/PS1 mice, and these include prostaglandin-endoperoxide synthases 1 and 2 (ptgs1 and 2) and nerve grow factor (ngf). We have also detected reduced expression of genes related to lipid metabolism in LDLr -/- mice: peroxisome proliferator activated receptor gamma (pparg), pro-opiomelanocortin-alpha (pomc) and of protein kinase, AMP-activated, alpha 1 catalytic subunit of AMPK (prkaa1). Our array data also indicate that transcriptional activity of early genes involved in memory process, such as FBJ osteosarcoma oncogene (Fos) and the activity regulated cytoskeletal-associated protein (Arc) gene, are increased in the hippocampus of LDLr -/- mice. Several proteins like insulin degrading enzyme (IDE), PGC-1α, OXPHOS 1, NMDAR1 and cyclic AMP response element binding protein (CREB) are up-regulated in the LDLr -/- mice, while in the APP/PS1 mouse model only OXPHOS complexes 2, 3 and 5 are slightly downregulated. Further studies are necessary to understand the molecular pathways involved in memory loss in hypercholesterolemic LDLr -/- mice.

Mice Lacking Functional Fas Death Receptors Are Protected from Kainic Acid-Induced Apoptosis in the Hippocampus.

The Fas receptor (FasR)/Fas ligand (FasL) system plays a significant role in the process of neuronal loss in neurological disorders. Thus, in the present study, we used a real-time PCR array focused apoptosis (Mouse Apoptosis RT(2) PCR Array) to study the role of the Fas pathway in the apoptotic process that occurs in a kainic acid (KA) mice experimental model. In fact, significant changes in the transcriptional activity of a total of 23 genes were found in the hippocampus of wild-type C57BL/6 mice after 12 h of KA treatment compared to untreated mice. Among the up-regulated genes, we found key factors involved in the extrinsic apoptotic pathway, such as tnf, fas and fasL, and also in caspase genes (caspase -4, caspase-8 and caspase-3). To discern the importance of the FasR/FasL pathway, mice lacking the functional Fas death receptor (lpr) were also treated with KA. After 24 h of neurotoxin treatment, lpr mice exhibited a reduced number of apoptotic positive cells, determined by the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) method in different regions of the hippocampus, when compared to wild-type mice. In addition, treatment of lpr mice with KA did not produce significant changes in the transcriptional activity of genes related to apoptosis in the hippocampus, either in the fas and fas ligand genes or in caspase-4 and caspase-8 and the executioner caspase-3 genes, as occurred in wild-type C57BL/6 mice. Thus, these data provide direct evidence that Fas signalling plays a key role in the induction of apoptosis in the hippocampus following KA treatment, making the inhibition of the death receptor pathway a potentially suitable target for excitotoxicity neuroprotection in neurological conditions such as epilepsy.

Treinamento (Entrenamiento) muscular inspiratório para o controle (el control) autonômico de indivíduos saudáveis (sanos) / Inspiratory muscle training in autonomic control in healthy individuals

Verificar o efeito (el efecto) agudo de diferentes sobrecargas de treinamento (entrenamiento) muscular inspiratório (TMI) sobre a (la) modulação autonômica cardiovascular em indivíduos saudáveis.Métodos: Ensaio (Ensayo) clínico randomizado cruzado. Foram (Fueron) incluídos voluntários saudáveis (sanos) entre 18 e 35 anos. A variabilidade da (de la) frequência cardíaca (VFC) foi analisada antes e após o (y después del) TMI a os (a los) 10 minutos, 60 minutos e 24 horas (agudo, subagudo e tardio, respectivamente). A força (La fuerza) muscular inspiratória foi avaliada (fue evaluada) através da (de la)medida de pressão inspiratória máxima (PImáx) com a utilização do (del) manovacuômetro digital MVD300 (Microhard System®, Globalmed, Porto Alegre, Brasil). O TMI foi realizado à 30% e 60% da PImáx ajustados no (en el) dispositivo pressórico linear Power breathe®. Resultados: Foram avaliados dezenove (diecinueve) indivíduos (47% homens, 25 ± 5 anos). Na (En la) fase aguda apenas com 60% da PImá houve redução (hubo una reducción) significativa da variabilidade dos (de los) intervalos RR e no (y enel) componente de alta frequência (HFnu), en quanto que o (mientras que el) componente de baixa(baja) frequência (LFnu) e o balanço (y el equilibrio) autonômico (LF/HF) aumentaram significativamente.Na (En la) fase subaguda, o mesmo comportamento foi observado para HFnu, LFnu, LF/HF. Quando comparadas as (Cuando se compararon las) sobrecargas, houve (hubo) aumento significativo na (en la) magnitude do efeito a 60% PImáx para NN50, LF/HF, LFnu, HFnu na fase aguda, bem como, para (así como para) RR, NN50, LFnu e HFnu na fase subaguda (p < 0.05). Conclusão: Agudamente, o efeito doTMI à 60% da PImáx foi maior deslocando a (fue más grande, desplazando la) modulação autonômicado sistema cardiovascular em indivíduos saudáveis para um predomínio simpático...

New insights into the immunological changes in IL-10-deficient mice during the course of spontaneous inflammation in the gut mucosa.

IL-10 is a regulatory cytokine that plays a major role in the homeostasis of the gut and this is illustrated by the fact that IL-10(-/-) mice develop spontaneous colitis. In this study, IL-10(-/-) mice were analyzed for immunological changes during colitis development. We found a reduced frequency of regulatory T cells CD4(+)CD25(+)Foxp3(+) and higher frequency of activated T cells in the colon that precedes the macroscopic signs of the disease. Production of IL-17 and IFN-γ was higher in the colon. Colitis progression culminates with the reduction of CD4(+)LAP(+) regulatory T cells in the intestine. Frequency of B1 cells and the secretory IgA production were both elevated. Despite these alterations, 16-week-old IL-10(-/-) mice could be rendered tolerant by a continuous feeding protocol. Our study provides detailed analysis of changes that precede colitis and it also suggests that oral tolerance could be used to design novel alternative therapies for the disease.

Gene expression profile in JNK3 null mice: a novel specific activation of the PI3K/AKT pathway.

JNK3 is mainly expressed in the CNS and it plays a crucial role in neuronal death in several neurodegenerative diseases. By contrast, the isoforms JNK1 and JNK2 seem to be involved in brain development. The lack of Jnk3 confers neuroprotection, although mechanisms responsible are unknown. The present study analyzes the gene expression profile in hippocampus from mice lacking Jnk3 in comparison to wild-type mice. The microarray analysis showed that 22 genes are differentially expressed (z-score>2 in two independent arrays) in Jnk3 null mice. Among these, we focused on pi3kcb, as it is directly related to the prosurvival phosphoinositide-3-kinase (PI3K)/AKT pathway. Results from Jnk3 null mice showed an increase in pik3cb transcript and protein, together with an increase in PI3K activity and phosphorylation of AKT. By contrast, these changes were not observed in Jnk1 null mice, which do not present neuroresistance to certain neurodegenerative insults. Therefore, our results indicate that the activation of PI3K/AKT pathway in hippocampus because of the increase in pik3cb transcription and that this mechanism is specifically related to the lack of Jnk3.

Content and traffic of taurine in hippocampal reactive astrocytes.

Taurine is one of the most abundant free amino acids in the mammalian central nervous system, where it is crucial to proper development. Moreover, taurine acts as a neuroprotectant in various diseases; in epilepsy, for example, it has the capacity to reduce or abolish seizures. In the present study, taurine levels has been determine in mice treated with Kainic Acid (KA) and results showed an increase of this amino acid in hippocampus but not in whole brain after 3 and 7 days of KA treatment. This increase occurs when gliosis was observed. Moreover, taurine transporter (TAUT) was found in astrocytes 3 and 7 days after KA treatment, together with an increase in cysteine sulfinic acid decarboxylase (csd) mRNA, that codifies for the rate-limiting enzyme of taurine synthesis, in the hippocampus at the same times after KA treatment. Glial cultures enriched in astrocytes were developed to demonstrate that these cells are responsible for changes in taurine levels after an injury to the brain. The cultures were treated with proinflammatory cytokines to reproduce gliosis. In this experimental model, an increase in the immunoreactivity of GFAP was observed, together with an increase in CSD and taurine levels. Moreover, an alteration in the taurine uptake-release kinetics was detected in glial cells treated with cytokine. All data obtained indicate that astrocytes could play a key role in taurine level changes induced by neuronal damage. More studies are, therefore, needed to clarify the role taurine has in relation to neuronal death and repair.