Marine Heatwave Caused Differentiated Dysbiosis in Photosymbiont Assemblages of Corals and Hydrocorals During El Niño 2015/2016.

Reef corals have been threatened by climate change, with more frequent and intense bleaching events leading to extensive coral mortality and loss of coral cover worldwide. In the face of this, the corals' photosymbiont assemblages have received special attention as a key to better understand the bleaching process and its recovery. To assess the effects of thermal anomalies, the coral Mussismilia harttii and the hydrocoral Millepora alcicornis were monitored through the El Niño 2015/2016 at a Southwestern Atlantic (SWA) coral reef. A severe bleaching event (57% of colonies bleached) was documented, triggered by a < 3 °C-week heatwave, but no mortality was detected. The hydrocoral was more susceptible than the scleractinian, displaying bleaching symptoms earlier and experiencing a longer and more intense bleaching event. The composition of photosymbionts in the M. alcicornis population was affected only at the rare biosphere level (< 5% relative abundance), with the emergence of new symbionts after bleaching. Conversely, a temporary dysbiosis was observed in the M. harttii population, with one of the dominant symbiodiniaceans decreasing in relative abundance at the peak of the bleaching, which negatively affected the total ß-diversity. After colonies' complete recovery, symbiodiniaceans' dominances returned to normal levels in both hosts. These results highlight critical differences in how the two coral species cope with bleaching and contribute to the understanding of the role of photosymbionts throughout the bleaching-recovery process.

Enamel erosion control by strontium-containing TiO2- and/or MgO-doped phosphate bioactive glass.

OBJECTIVES: To evaluate the effect of strontium-containing titanium- and/or magnesium-doped phosphate bioactive glass on the control of dental erosion. MATERIALS AND METHODS: Fifty fragments of human enamel were divided into five groups: negative control, 45S5 bioglass, strontium-containing Ti-doped phosphate bioactive glass (PBG-Ti), strontium-containing Mg-doped phosphate bioactive glass (PBG-Mg), and strontium-containing Ti- and Mg-doped phosphate bioactive glass (PBG-TiMg). The specimens underwent cycles of erosive challenge twice daily for 5 days with 1 mL of citric acid for 2 min followed by 1 mL of the suspension with bioactive substances for 3 min. After the cycles, profilometry, roughness and microhardness testing, and scanning electron microscopy (SEM) were performed. The following statistical tests were used: one-way ANOVA (profile, roughness, and surface microhardness (%VMS) data variation), Tukey's HSD (%VMS), Games-Howell test (profilometry), Student's t test (roughness), and Pearson's correlation between the variables. RESULTS: The lower loss of enamel surface and lower %VMS was observed in the PBG-Mg and PBG-TiMg groups, and only the PBG-Mg group showed similar roughness between baseline and eroded areas (p > 0.05). On SEM micrographs, PBG-Ti and PBG-Mg groups showed lower apparent demineralization. CONCLUSION: All bioactive materials protected the enamel against erosion. However, strontium-containing phosphate bioactive glasses showed lower enamel loss, and the presence of Mg in these bioactive glasses provided a greater protective effect. CLINICAL RELEVANCE: Experimental strontium-containing phosphate bioactive glasses are effective in controlling enamel erosion. The results obtained in this study will guide the development of new dental products.

Ten years of iPSC: clinical potential and advances in vitro hematopoietic differentiation.

Ten years have passed since the first publication announcing the generation of induced pluripotent stem cells (iPSCs). Issues related to ethics, immune rejection, and cell availability seemed to be solved following this breakthrough. The development of iPSC technology allows advances in in vitro cell differentiation for cell therapy purpose and other clinical applications. This review provides a perspective on the iPSC potential for cell therapies, particularly for hematological applications. We discuss the advances in in vitro hematopoietic differentiation, the possibilities to employ iPSC in hematology studies, and their potential clinical application in hematologic diseases. The generation of red blood cells and functional T cells and the genome editing technology applied to mutation correction are also covered. We highlight some of the requirements and obstacles to be overcome before translating these cells from research to the clinic, for instance, iPSC variability, genotoxicity, the differentiation process, and engraftment. Also, we evaluate the patent landscape and compile the clinical trials in the field of pluripotent stem cells. Currently, we know much more about iPSC than in 2006, but there are still challenges that must be solved. A greater understanding of molecular mechanisms underlying the generation of hematopoietic stem cells is necessary to produce suitable and transplantable hematopoietic stem progenitor cells from iPSC.

Multiple events determine the distribution of fishes' early life stages in the Abrolhos Bank, Southwestern Atlantic.

Fluctuations in reef fish settlement and recruitment significantly impact local population dynamics and adult population size. We use light traps to evaluate reef fish settlement in inshore and offshore reefs in the Abrolhos Bank, in addition to the spatio-temporal distribution patterns, and environmental variables' influence. Data were collected from inshore and offshore reefs, and statistical analyses, including GLMs, PERMANOVA and nMDS, aimed to understand spatial-temporal species distribution patterns and the relationship between environmental variables. Results indicate greater species abundance and diversity offshore, influenced by winds, tides, and distance from shore, explaining 50% of larval patterns and highlighting the importance of spatial-temporal variations in early-life reef fish assemblages. However, several factors, including habitat type, quality, complexity, seascape configuration, and MPA location, may influence these patterns. Additionally, we provide evidence that commercially important species like Lutjanids use Abrolhos reef areas as nursery grounds.

Coupling remote sensing and eDNA to monitor environmental impact: A pilot to quantify the environmental benefits of sustainable agriculture in the Brazilian Amazon.

Monitoring is essential to ensure that environmental goals are being achieved, including those of sustainable agriculture. Growing interest in environmental monitoring provides an opportunity to improve monitoring practices. Approaches that directly monitor land cover change and biodiversity annually by coupling the wall-to-wall coverage from remote sensing and the site-specific community composition from environmental DNA (eDNA) can provide timely, relevant results for parties interested in the success of sustainable agricultural practices. To ensure that the measured impacts are due to the environmental projects and not exogenous factors, sites where projects have been implemented should be benchmarked against counterfactuals (no project) and control (natural habitat) sites. Results can then be used to calculate diverse sets of indicators customized to monitor different projects. Here, we report on our experience developing and applying one such approach to assess the impact of shaded cocoa projects implemented by the Instituto de Manejo e Certificação Florestal e Agrícola (IMAFLORA) near São Félix do Xingu, in Pará, Brazil. We used the Continuous Degradation Detection (CODED) and LandTrendr algorithms to create a remote sensing-based assessment of forest disturbance and regeneration, estimate carbon sequestration, and changes in essential habitats. We coupled these remote sensing methods with eDNA analyses using arthropod-targeted primers by collecting soil samples from intervention and counterfactual pasture field sites and a control secondary forest. We used a custom set of indicators from the pilot application of a coupled monitoring framework called TerraBio. Our results suggest that, due to IMAFLORA's shaded cocoa projects, over 400 acres were restored in the intervention area and the community composition of arthropods in shaded cocoa is closer to second-growth forests than that of pastures. In reviewing the coupled approach, we found multiple aspects worked well, and we conclude by presenting multiple lessons learned.

Emerging CAR T cell therapies: clinical landscape and patent technological routes.

The purpose of this study is to mine CAR-T patents and therapies under development, to design a landscape of the sector and to understand key therapy segments and their current trends. The study analyzed the entire market, consisting of 1624 patent families and 509 biologics under development, to depict an overview of the CAR-T therapies and their state of the art. Our results showed cutting-edge inventions, the major players, the dynamics of cooperation among institutions, the progress of the therapies' generation over the years and future innovation pathways. CAR-T therapies are transforming the current scenario for cancer treatment, and this study reveals the picture of what we can likely expect ahead in order to assist scientists at the academy and industry to improve their research strategies.

Larvae of the South Atlantic coral Favia gravida are tolerant to salinity and nutrient concentrations associated with river discharges.

Rivers release freshwater, nutrients and pollutants into reefs. This type of environmental stress reduces coral larvae settlement and alter its energy metabolism. We investigated the tolerance of Favia gravida (Scleractinia) larvae to river discharges. We exposed larvae to (i) different salinities (25, 30, 35 and 40 PSU); and (ii) dilutions of river water containing nutrients and metals (0, 20, 40, 60, 80 and 100% river water) under control salinity of 35 PSU. We then examined settlement and larval enzymatic activity. No differences in settlement were detected among salinities. Settlement was also similar to control for larvae under 100% river water. Enzymatic activity for citrate synthase remained unaltered for all treatments. Lactate dehydrogenase activity was slightly altered under different salinities, suggesting a mild stress response. Findings suggest that F. gravida larvae are tolerant to a wide range of salinity and nutrient conditions and that this is a stress-tolerant species.

Vallissiana universitaria (Lepidoptera: Gracillariidae): a new genus and species of leaf-mining moth associated with Erythroxylum (Erythroxylaceae) in the Atlantic Forest of Brazil.

Vallissiana universitaria Pereira Arévalo, a new genus and species of leaf-miner moth (Gracillariidae: Gracillariinae) is described and illustrated with the aid of optical and scanning electron microscopy, including adults, larva, pupa and the mine. Its monophyletic status is confirmed within the subfamily based on a DNA barcode CoI tree. The immature stages are associated with Erythroxylum argentinum O. E. Schulz (Erythroxylaceae) and four larval instars are found, all forming a round blotch mine from the beginning of ontogeny. The first two instars are sap-feeders, using only the epidermal cells, whereas the last two are tissue-feeders, mining the parenchyma cells. Pupation occurs inside the leaf mine within a flimsy, silk-made cocoon. This is the third endemic genus of gracillariid moths described from the Atlantic Forest of Brazil and the first associated with Erythroxylum P. Browne. Characteristics found on the forewing and in the last abdominal segments of the adult were determinant for the proposition of the new genus. The CoI tree indicated that it is closely related to Aspilapteryx, while this genus was recovered as polyphyletic in the analyses. Morphological evidence supports this polyphyly. Consequently, Sabulopteryx Triberti, 1985, stat. nov. is considered a valid genus.

The lncRNA RMEL3 protects immortalized cells from serum withdrawal-induced growth arrest and promotes melanoma cell proliferation and tumor growth.

RMEL3 is a recently identified lncRNA associated with BRAFV600E mutation and melanoma cell survival. Here, we demonstrate strong and moderate RMEL3 upregulation in BRAF and NRAS mutant melanoma cells, respectively, compared to melanocytes. High expression is also more frequent in cutaneous than in acral/mucosal melanomas, and analysis of an ICGC melanoma dataset showed that mutations in RMEL3 locus are preponderantly C > T substitutions at dipyrimidine sites including CC > TT, typical of UV signature. RMEL3 mutation does not correlate with RMEL3 levels, but does with poor patient survival, in TCGA melanoma dataset. Accordingly, RMEL3 lncRNA levels were significantly reduced in BRAFV600E melanoma cells upon treatment with BRAF or MEK inhibitors, supporting the notion that BRAF-MEK-ERK pathway plays a role to activate RMEL3 gene transcription. RMEL3 overexpression, in immortalized fibroblasts and melanoma cells, increased proliferation and survival under serum starvation, clonogenic ability, and xenografted melanoma tumor growth. Although future studies will be needed to elucidate the mechanistic activities of RMEL3, our data demonstrate that its overexpression bypasses the need of mitogen activation to sustain proliferation/survival of non-transformed cells and suggest an oncogenic role for RMEL3.

Environmental health in southwestern Atlantic coral reefs: Geochemical, water quality and ecological indicators.

Climate change, pollution and increased runoff are some of the main drivers of coral reefs degradation worldwide. However, the occurrence of runoff and marine pollution, as well as its ecological effects in South Atlantic coral reefs are still poorly understood. The aim of the present work is to characterize the terrigenous influence and contamination impact on the environmental health of five reefs located along a gradient of distance from a river source, using geochemical, water quality, and ecological indicators. Stable isotopes and sterols were used as geochemical indicators of sewage and terrigenous organic matter. Dissolved metal concentrations (Cu, Zn, Cd, and Pb) were used as indicators of water quality. Population density, bleaching and chlorophyll α content of the symbiont-bearing foraminifer Amphistegina gibbosa, were used as indicators of ecological effects. Sampling was performed four times during the year to assess temporal variability. Sediment and water quality indicators showed that reefs close to the river discharge experience nutrient enrichment and sewage contamination, and metals concentrations above international environmental quality guidelines. Higher levels of contamination were strongly related to the higher frequency of bleaching and lower density in A. gibbosa populations. The integrated evaluation of stable isotopes, sterols and metals provided a consistent diagnostic about sewage influence on the studied reefs. Additionally, the observed bioindicator responses evidenced relevant ecological effects. The water quality, geochemical and ecological indicators employed in the present study were effective as biomonitoring tools to be applied in reefs worldwide.

Uncovering Innovation Features and Emerging Technologies in Molecular Biology through Patent Analysis.

Scientific research at universities has a crucial role in leveraging a country's innovative potential. Sectors that require greater investments in technology for the development of their research, such as biotechnology, need to be aware of the frontier state-of-the-art technology and the knowledge incrusted within it. Although the information available in scientific articles is well explored in academic environment, the patent literature, where much of the technological information is present, is still poorly accessed. This chapter is intended to instruct students and researchers at universities to look at patent document analysis as a source of scientific and technological information and explore its applications. Within this chapter, we use the technological area regarding immunoglobulins inventions (monoclonal and polyclonal antibodies) as example to provide directions on how to develop a patent landscape to get an overview of the inventions in a certain field; how to map a collaborative network of inventors/assignees to help the pursuit and identification of future partnerships; and lastly we describe the steps of how to set up a network of patent citations with the aim of forecasting emerging technologies. We strongly believe that incorporate data from patents in planning phase of research projects at academia, as well as to establish partnerships and join R&D efforts to invest on promising technologies, is of great relevance to leverage the growth of the biotechnology sector.

Copper effects on biomarkers associated with photosynthesis, oxidative status and calcification in the Brazilian coral Mussismilia harttii (Scleractinia, Mussidae).

Seawater contamination with metals, such as copper (Cu), is a notable local impact threatening coral reefs. Cu effects on biomarkers associated with photosynthesis, oxidative status and calcification were evaluated in the Brazilian coral Mussismilia harttii using a marine mesocosm facility. Polyps were kept under control conditions (1.9 µg L-1 Cu) or exposed to dissolved Cu (3.0, 4.8, and 6.7 µg L-1) for 12 days. Photochemical efficiency of the photosystem II of symbiotic algae (zooxanthellae) was measured and polyps were analyzed for antioxidant capacity, lipid peroxidation, DNA damage, and carbonic anhydrase Ca-ATPase, Mg-ATPase and (Ca,Mg)-ATPase activities after 12 days. Results highlighted the effects of Cu exposure, leading corals to an oxidative stress condition [increased total antioxidant capacity (TAC) and DNA damage] and a possible reduced calcification ability [decreased (Ca,Mg)-ATPase activity]. Therefore, biomarkers associated with oxidative status (TAC and DNA damage) and calcification [(Ca, Mg)-ATPase] are indicated as good predictors of corals health.

Effects of Ocean Acidification and Temperature Increases on the Photosynthesis of Tropical Reef Calcified Macroalgae.

Climate change is a global phenomenon that is considered an important threat to marine ecosystems. Ocean acidification and increased seawater temperatures are among the consequences of this phenomenon. The comprehension of the effects of these alterations on marine organisms, in particular on calcified macroalgae, is still modest despite its great importance. There are evidences that macroalgae inhabiting highly variable environments are relatively resilient to such changes. Thus, the aim of this study was to evaluate experimentally the effects of CO2-driven ocean acidification and temperature rises on the photosynthesis of calcified macroalgae inhabiting the intertidal region, a highly variable environment. The experiments were performed in a reef mesocosm in a tropical region on the Brazilian coast, using three species of frondose calcifying macroalgae (Halimeda cuneata, Padina gymnospora, and Tricleocarpa cylindrica) and crustose coralline algae. The acidification experiment consisted of three treatments with pH levels below those occurring in the region (-0.3, -0.6, -0.9). For the temperature experiment, three temperature levels above those occurring naturally in the region (+1, +2, +4°C) were determined. The results of the acidification experiment indicate an increase on the optimum quantum yield by T. cylindrica and a decline of this parameter by coralline algae, although both only occurred at the extreme acidification treatment (-0.9). The energy dissipation mechanisms of these algae were also altered at this extreme condition. Significant effects of the temperature experiment were limited to an enhancement of the photosynthetic performance by H. cuneata although only at a modest temperature increase (+1°C). In general, the results indicate a possible photosynthetic adaptation and/or acclimation of the studied macroalgae to the expected future ocean acidification and temperature rises, as separate factors. Such relative resilience may be a result of the highly variable environment they inhabit.

RMEL3, a novel BRAFV600E-associated long noncoding RNA, is required for MAPK and PI3K signaling in melanoma.

Previous work identified RMEL3 as a lncRNA with enriched expression in melanoma. Analysis of The Cancer Genome Atlas (TCGA) data confirmed RMEL3 enriched expression in melanoma and demonstrated its association with the presence of BRAFV600E. RMEL3 siRNA-mediated silencing markedly reduced (95%) colony formation in different BRAFV600E melanoma cell lines. Multiple genes of the MAPK and PI3K pathways found to be correlated with RMEL3 in TCGA samples were experimentally confirmed. RMEL3 knockdown led to downregulation of activators or effectors of these pathways, including FGF2, FGF3, DUSP6, ITGB3 and GNG2. RMEL3 knockdown induces gain of protein levels of tumor suppressor PTEN and the G1/S cyclin-Cdk inhibitors p21 and p27, as well as a decrease of pAKT (T308), BRAF, pRB (S807, S811) and cyclin B1. Consistently, knockdown resulted in an accumulation of cells in G1 phase and subG0/G1 in an asynchronously growing population. Thus, TCGA data and functional experiments demonstrate that RMEL3 is required for MAPK and PI3K signaling, and its knockdown decrease BRAFV600E melanoma cell survival and proliferation.

Beneficial Effect of Synbiotic Supplementation on Hepatic Steatosis and Anthropometric Parameters, But Not on Gut Permeability in a Population with Nonalcoholic Steatohepatitis.

Nonalcoholic fatty liver disease is the most prevalent chronic liver disease in Western countries; it can progress to nonalcoholic steatohepatitis (NASH), cirrhosis and hepatocarcinoma. The importance of gut-liver-adipose tissue axis has become evident and treatments targeting gut microbiota may improve inflammatory and metabolic parameters in NASH patients. In a randomized, controlled clinical trial, involving 50 biopsy-proven NASH patients, we investigated the effects of synbiotic supplementation on metabolic parameters, hepatic steatosis, intestinal permeability, small intestinal bacterial overgrowth (SIBO) and lipopolysaccharide (LPS) serum levels. Patients were separated into two groups receiving Lactobacillus reuteri with guar gum and inulin for three months and healthy balanced nutritional counseling versus nutritional counseling alone. Before and after the intervention we assessed steatosis by magnetic resonance imaging, intestinal permeability by lactulose/mannitol urinary excretion and SIBO by glucose breath testing. NASH patients presented high gut permeability, but low prevalence of SIBO. After the intervention, only the synbiotic group presented a reduction in steatosis, lost weight, diminished BMI and waist circumference measurement. Synbiotic did not improve intestinal permeability or LPS levels. We concluded that synbiotic supplementation associated with nutritional counseling seems superior to nutritional counseling alone for NASH treatment as it attenuates steatosis and may help to achieve weight loss.

A novel marine mesocosm facility to study global warming, water quality, and ocean acidification.

We describe a completely randomizable flow-through outdoor mesocosm for climate change and ecotoxicology studies that was built with inexpensive materials. The 16 raceway tanks allow up to 6× water renewal per hour, avoiding changes in natural abiotic seawater conditions. We use an open-source hardware board (Arduino) that was adapted to control heaters and an innovative CO 2 injection system. This system reduced seawater pH up to -0.9 units and increased temperature up to +6°C in three treatments and a control. Treatments can be continuously compared with the control and vary according to diel fluctuations, thus following the diel range observed in the sea. The mesocosm facility also includes an integrated secondary system of 48 aquaria for ecotoxicology studies. We validated the reproducibility and relevance of our experimental system by analyzing the variation of the total DNA of the microbial community extracted from corals in three elevated temperature scenarios during a 40-day experiment. We also present data from temperature, acidification, and copper contamination trials, which allowed continuous, reliable, and consistent treatment manipulations.

The involvement of heparin in retinal infection by Toxoplasma gondii in a chick model revealed an ontogenetic-dependent pattern.

This work aimed to test the influence of heparin on the susceptibility of retinal cells to Toxoplasma gondii infection. Primary cultures of retinas from chick embryos of 8 (E8) or 11 (E11) days and fibroblasts (control) were used. To determine the influence of heparin in T. gondii infection, tachyzoites of the RH strain were treated with heparin before addition in the culture. A monoclonal anti-heparin antibody was used to analyze the heparin distribution on fibroblast and retinal cell surfaces. Our results showed that retinal cells (E8 and E11) had a higher infection rate than fibroblasts (91% and 24% versus 13%, respectively). Pre-treatment of T. gondii with heparin decreased infection of E8 retinal cells when compared with non-treated parasites (45% versus 91%, respectively), but not of E11 cells (35% versus 48%). In accordance, retinal cells presented an intense heparin staining by immunofluorescence assay. In conclusion, retinal cells from chick embryos were more susceptible to infection by T. gondii compared to fibroblasts and, pre-treatment of tachyzoites with heparin decreased the number of infected cells and parasite burden particularly for E8 retinal cells.

Molecular profiling of patient-matched brain and extracranial melanoma metastases implicates the PI3K pathway as a therapeutic target.

PURPOSE: An improved understanding of the molecular pathogenesis of brain metastases, one of the most common and devastating complications of advanced melanoma, may identify and prioritize rational therapeutic approaches for this disease. In particular, the identification of molecular differences between brain and extracranial metastases would support the need for the development of organ-specific therapeutic approaches. EXPERIMENTAL DESIGN: Hotspot mutations, copy number variations (CNV), global mRNA expression patterns, and quantitative analysis of protein expression and activation by reverse-phase protein array (RPPA) analysis were evaluated in pairs of melanoma brain metastases and extracranial metastases from patients who had undergone surgical resection for both types of tumors. RESULTS: The status of 154 previously reported hotspot mutations, including driver mutations in BRAF and NRAS, were concordant in all evaluable patient-matched pairs of tumors. Overall patterns of CNV, mRNA expression, and protein expression were largely similar between the paired samples for individual patients. However, brain metastases demonstrated increased expression of several activation-specific protein markers in the PI3K/AKT pathway compared with the extracranial metastases. CONCLUSIONS: These results add to the understanding of the molecular characteristics of melanoma brain metastases and support the rationale for additional testing of the PI3K/AKT pathway as a therapeutic target in these highly aggressive tumors.

Inhibition of mTORC1/2 overcomes resistance to MAPK pathway inhibitors mediated by PGC1α and oxidative phosphorylation in melanoma.

Metabolic heterogeneity is a key factor in cancer pathogenesis. We found that a subset of BRAF- and NRAS-mutant human melanomas resistant to the MEK inhibitor selumetinib displayed increased oxidative phosphorylation (OxPhos) mediated by the transcriptional coactivator PGC1α. Notably, all selumetinib-resistant cells with elevated OxPhos could be resensitized by cotreatment with the mTORC1/2 inhibitor AZD8055, whereas this combination was ineffective in resistant cell lines with low OxPhos. In both BRAF- and NRAS-mutant melanoma cells, MEK inhibition increased MITF expression, which in turn elevated levels of PGC1α. In contrast, mTORC1/2 inhibition triggered cytoplasmic localization of MITF, decreasing PGC1α expression and inhibiting OxPhos. Analysis of tumor biopsies from patients with BRAF-mutant melanoma progressing on BRAF inhibitor ± MEK inhibitor revealed that PGC1α levels were elevated in approximately half of the resistant tumors. Overall, our findings highlight the significance of OxPhos in melanoma and suggest that combined targeting of the MAPK and mTORC pathways may offer an effective therapeutic strategy to treat melanomas with this metabolic phenotype.