Microplastic pollution in water, sediments and commercial fish species from Ciénaga Grande de Santa Marta lagoon complex, Colombian Caribbean.

Microplastics are emerging pollutants that have been found in different environmental matrices of marine and coastal ecosystems, where they can generate harmful ecological impacts. Little is known about the current state of microplastic pollution in fragile tropical lagoon ecosystems, such as Ciénaga Grande de Santa Marta (CGSM) in the Caribbean coast of Colombia. This study assesses microplastic pollution in surface waters and sediments, and the occurrence of microplastic ingestion in commercially important fish species from CGSM. In waters, microplastic abundances ranged from 0.0 to 0.3 items L-1 while in sediments they varied from 0.0 to 3.1 items kg-1. The most abundant types of microplastics are fibers and fragments, with polypropylene, polyethylene and high-density polyethylene as the most abundant polymers. Also, 100 (i.e. 21.1%) out of 474 individuals from nine fish species had microplastics in their digestive tracts. Microplastics present in water and sediments and in the digestive tract of the analyzed fish species have similar characteristics, also showing a moderate and statistically significant association. Microplastic abundances are higher near river mouths and in urban areas with a high density of fishing activities and aquaculture infrastructures, which are important sources of contaminants. Microplastic pollution in CGSM represents a threat to the lagoon ecosystem and to local people depending on artisanal fishing. Consequently, effective actions to reduce pollution and its socio-environmental impacts are urgently required.

In vitro biological activity of extracts from marine bacteria cultures against Toxoplasma gondii and Mycobacterium tuberculosis.

AIMS: To evaluate the biological activity of extracts from cultures of marine bacteria against Toxoplasma gondii and Mycobacterium tuberculosis. METHODS AND RESULTS: Ethyl acetate extracts obtained from seven marine bacteria were tested against T. gondii GFP-RH and M. tuberculosis H37Rv. The cytotoxicity on HFF-1 cells was measured by a microplate resazurin fluorescent approach, and the haemolytic activity was determined photometrically. The extracts from Bacillus sp. (INV FIR35 and INV FIR48) affected the tachyzoite viability. The extracts from Bacillus, Pseudoalteromonas, Streptomyces and Micromonospora exhibited effects on infection and proliferation processes of parasite. Bacillus sp. INV FIR48 extract showed an minimum inhibitory concentration value of 50 µg ml-1 against M. tuberculosis H37Rv. All the extracts exhibited relatively low toxicity to HFF-1 cells and the primary culture of erythrocytes, except Bacillus sp. INV FIR35, which decreased cell viability under 20%. Liquid chromatography coupled to mass spectrometry analysis of the most active bacterial extract Bacillus sp. INV FIR48 showed the presence of peptide metabolites related to surfactin. CONCLUSIONS: The extract from culture of deep-sea Bacillus sp. INV FIR48 showed anti-T. gondii and anti-tuberculosis (TB) biological activity with low cytotoxicity. In addition, peptide metabolites were detected in the extract. SIGNIFICANCE AND IMPACT OF THE STUDY: Toxoplasmosis and TB are among the most prevalent diseases worldwide, and the current treatment drugs exhibit side effects. This study confirm that marine bacteria are on hand sources of anti-infective natural products.

Dereplication of antimicrobial biosurfactants from marine bacteria using molecular networking.

Biosurfactants are amphiphilic surface-active molecules of microbial origin principally produced by hydrocarbon-degrading bacteria; in addition to the bioremediation properties, they can also present antimicrobial activity. The present study highlights the chemical characterization and the antimicrobial activities of biosurfactants produced by deep-sea marine bacteria from the genera Halomonas, Bacillus, Streptomyces, and Pseudomonas. The biosurfactants were extracted and chemically characterized through Chromatography TLC, FT-IR, LC/ESI-MS/MS, and a metabolic analysis was done through molecular networking. Six biosurfactants were identified by dereplication tools from GNPS and some surfactin isoforms were identified by molecular networking. The half-maximal inhibitory concentration (IC50) of biosurfactant from Halomonas sp. INV PRT125 (7.27 mg L-1) and Halomonas sp. INV PRT124 (8.92 mg L-1) were most effective against the pathogenic yeast Candida albicans ATCC 10231. For Methicillin-resistant Staphylococcus aureus ATCC 43300, the biosurfactant from Bacillus sp. INV FIR48 was the most effective with IC50 values of 25.65 mg L-1 and 21.54 mg L-1 for C. albicans, without hemolytic effect (< 1%), and non-ecotoxic effect in brine shrimp larvae (Artemia franciscana), with values under 150 mg L-1, being a biosurfactant promising for further study. The extreme environments as deep-sea can be an important source for the isolation of new biosurfactants-producing microorganisms with environmental and pharmaceutical use.

Changing the Paradigm of Craniofacial Reconstruction: A Prospective Clinical Trial of Autologous Fat Transfer for Craniofacial Deformities.

OBJECTIVE: This study aimed to prospectively assess outcomes for surgical autologous fat transfer (AFT) applied for traumatic and postsurgical craniofacial deformities. The minimally invasive nature of AFT has potential for reduced risk and superior outcomes compared with current reconstructive options. BACKGROUND: Craniofacial deformities have functional and psychosocial sequelae and can profoundly affect quality of life. Traditional reconstructive options are invasive, invasive, complex, and often lack precision in outcomes. Although AFT is safe, effective, and minimally invasive, only anecdotal evidence exists for reconstruction of craniofacial deformities. METHODS: In this Institutional Review Board-approved prospective cohort study, 20 subjects underwent AFT (average volume: 23.9 ±â€Š13.2 mL). Volume retention over time was determined using high-resolution computed tomography. Flow cytometry was used to assess cellular subpopulations and viability in the stromal vascular fraction. Quality of life assessments were performed. After the completion of 9-month follow-up, 5 subjects were enrolled for a second treatment. RESULTS: No serious adverse events occurred. Volume retention averaged 63 ±â€Š17% at 9 months. Three-month retention strongly predicted 9-month retention (r=0.996, P < 0.0001). There was no correlation between the total volume injected and retention. Patients undergoing a second procedure had similar volume retention as the first (P = 0.05). Age, sex, body mass index, and stromal vascular fraction cellular composition did not impact retention. Surprisingly, former smokers had greater volume retention at 9 months compared with nonsmokers (74.4% vs 56.2%, P = 0.009). Satisfaction with physical appearance (P = 0.002), social relationships (P = 0.02), and social functioning quality of life (P = 0.05) improved from baseline to 9 months. CONCLUSIONS: For craniofacial defects, AFT is less invasive and safer than traditional reconstructive options. It is effective, predictable, and reaches volume stability at 3 months. Patient-reported outcomes demonstrate a positive life-changing impact.

Antibody-based cell-surface proteome profiling of metastatic breast cancer primary explants and cell lines.

Flow cytometric cell surface proteomics provides a new and powerful tool to determine changes accompanying neoplastic transformation and invasion, providing clues to essential interactions with the microenvironment as well as leads for potential therapeutic targets. One of the most important advantages of flow cytometric cell surface proteomics is that it can be performed on living cells that can be sorted for further characterization and functional studies. Here, we document the surface proteome of clonogenic metastatic breast cancer (MBrCa) explants, which was strikingly similar to that of normal mesenchymal stromal cells (P = 0.017, associated with Pearson correlation coefficient) and transformed mammary epithelial cells (P = 0.022). Markers specifically upregulated on MBrCa included CD200 (Ox2), CD51/CD61 (Integrin α5/ß3), CD26 (dipeptidyl peptidase-4), CD165 (c-Cbl), and CD54 (ICAM-1). Proteins progressively upregulated in a model of neoplastic transformation and invasion included CD26, CD63 (LAMP3), CD105 (Endoglin), CD107a (LAMP1), CD108 (Semaphorin 7A), CD109 (Integrin ß4), CD151 (Raph blood group), and disialoganglioside G2. The proteome of the commonly used cell lines MDA-MB-231, MCF7, and BT-474 were uncorrelated with that of MBrCa (P = 1.0, 1.0, 0.9, respectively). The comparison has demonstrated the mesenchymal nature of clonogenic cells isolated by short-term culture of metastatic breast cancer, provided several leads for biomarkers and potential targets for anti-invasive therapy, including CD200, and highlighted the limitations of breast cancer cell lines for representing the cell surface biology of breast cancer. © 2017 International Society for Advancement of Cytometry.

Hyperferritinemia is a potential marker of chronic chikungunya: A retrospective study on the Island of Curaçao during the 2014-2015 outbreak.

BACKGROUND: Recently Chikungunya virus (CHIKV) outbreaks have been reported in the Carribean. There is no data regarding the outbreak in Curaçao. In addition, to date there is no biomarker that could be used to predict chronic infection. OBJECTIVES: To characterize the first CHIKV outbreak in Curaçao and to identify potential biomarkers for chronic infection. STUDY DESIGN: A serological test and quantitative polymerase chain reaction (qPCR) were used on samples collected in Curaçao to confirm infection. Subsequently, six samples with high viral load were selected for phylogenetic analysis. Furthermore we investigated the association of macrophage-related biomarkers during CHIKV infection with chronic arthralgia/arthritis. RESULTS: 116 patients in Curacao were diagnosed with CHIKV infection based on ELISA and 77% were tested positive for CHIKV by qPCR. Phylogenetic analysis showed that an Asian genotype was the cause of the outbreak. Elevated levels of ferritin and CRP were significantly associated with viraemia. In addition, elevated ferritin levels were significantly associated with chronic arthralgia. CONCLUSIONS: The results showed that the presence of an Asian genotype of CHIKV in Curaçao for the first time. Moreover, we found an association between ferritin levels with chronic arthralgia.

Hyperferritinaemia in dengue virus infected patients is associated with immune activation and coagulation disturbances.

BACKGROUND: During a dengue outbreak on the Caribbean island Aruba, highly elevated levels of ferritin were detected in dengue virus infected patients. Ferritin is an acute-phase reactant and hyperferritinaemia is a hallmark of diseases caused by extensive immune activation, such as haemophagocytic lymphohistiocytosis. The aim of this study was to investigate whether hyperferritinaemia in dengue patients was associated with clinical markers of extensive immune activation and coagulation disturbances. METHODOLOGY/PRINCIPAL FINDINGS: Levels of ferritin, standard laboratory markers, sIL-2R, IL-18 and coagulation and fibrinolytic markers were determined in samples from patients with uncomplicated dengue in Aruba. Levels of ferritin were significantly increased in dengue patients compared to patients with other febrile illnesses. Moreover, levels of ferritin associated significantly with the occurrence of viraemia. Hyperferritinaemia was also significantly associated with thrombocytopenia, elevated liver enzymes and coagulation disturbances. The results were validated in a cohort of dengue virus infected patients in Brazil. In this cohort levels of ferritin and cytokine profiles were determined. Increased levels of ferritin in dengue virus infected patients in Brazil were associated with disease severity and a pro-inflammatory cytokine profile. CONCLUSIONS/SIGNIFICANCE: Altogether, we provide evidence that ferritin can be used as a clinical marker to discriminate between dengue and other febrile illnesses. The occurrence of hyperferritinaemia in dengue virus infected patients is indicative for highly active disease resulting in immune activation and coagulation disturbances. Therefore, we recommend that patients with hyperferritinaemia are monitored carefully.

Serum angiopoietin-2 and soluble VEGF receptor 2 are surrogate markers for plasma leakage in patients with acute dengue virus infection.

BACKGROUND: Endothelial cell dysfunction is believed to play an important role in the pathogenesis of plasma leakage in patients with acute dengue virus (DENV) infection. Several factors, produced by activated endothelial cells, have been associated with plasma leakage or severe disease in patients with infectious diseases. OBJECTIVES: The aim of this study was to investigate which of these markers could serve as a surrogate marker for the occurrence of plasma leakage in patients with acute DENV infection. STUDY DESIGN: A case-control study was performed in patients with acute DENV infection in Santos, Brazil. Plasma leakage was detected with X-ray and/or ultrasound examination at admission. Serum levels of soluble endoglin, endothelin-1, angiopoietin-2, VEGF, soluble VEGFR-2, MMP-2, MMP-9, TIMP-1 and TIMP-2 were determined using commercially available ELISAs. RESULTS: Increased levels of angiopoietin-2, endothelin-1 and MMP-2 and decreased levels of soluble VEGFR-2 were significantly associated with the occurrence of plasma leakage. An unsupervised cluster analysis confirmed that angiopoietin-2 and soluble VEGFR-2 were strongly associated with clinical apparent vascular leakage. CONCLUSION: Angiopoietin-2 and soluble VEGFR-2 can serve as surrogate markers for the occurrence of plasma leakage in patients with acute DENV infection.

CD34 affinity pheresis attenuates a surge among circulating progenitor cells following vascular injury.

BACKGROUND: Intimal hyperplasia (restenosis) is an exaggerated healing response leading to failure of half of vascular interventions. Increasing evidence suggests that circulating progenitor cells contribute to intimal pathology, and clinical studies have demonstrated a correlation between progenitor cells and the incidence of restenosis after cardiovascular interventions. The aims of this study were to characterize the temporal response of CD34+ progenitors following vascular injury in an ovine model and to evaluate an affinity pheresis approach to attenuate this response. METHODS: An ovine model underwent either operative vascular injury or a nonvascular surgery (n = 3 per group). Blood was examined perioperatively over 2 weeks by flow cytometry. Next, an affinity pheresis approach to mediate systemic depletion of CD34 progenitors was designed. Custom agarose pheresis matrix with antibody affinity toward CD34 or an isotype control was evaluated in vitro. Next, following vascular injury, sheep underwent perioperative whole blood volume pheresis toward either the progenitor cell marker CD34 (n = 3) or an isotype control (n = 4) for 14 days. Animals were monitored by physical exam as well as complete blood counts. Cells recovered by pheresis were eluted and examined by flow cytometry. RESULTS: Flow cytometry revealed a focal surge of circulating CD34 cells after vascular injury but not among surgical controls (P = .05). Toward the goal of an approach to attenuate the surge of CD34 progenitors, an evaluation of high-flow affinity matrix revealed efficacy in removal of progenitors from ovine blood in vitro. Next, a separate group of animals undergoing affinity pheresis after vascular injury was evaluated to mediate systemic depletion of CD34+ cells. Again, a surge of CD34+ cells was observed among isotype pheresis animals following vascular intervention but was attenuated over 20-fold by a CD34 pheresis approach (P = .029). Furthermore, an average of 77 million CD34-positive cells were eluted from the CD34 pheresis matrix. Despite multiple sessions of pheresis, complete blood counts remained essentially unchanged over 2 weeks. CONCLUSIONS: Despite evidence suggesting a role for CD34+ circulating progenitor cells in restenotic pathology, the temporal pattern of CD34 progenitors after vascular injury has not been previously defined. We have demonstrated a surge among circulating CD34+ cells that appears confined to procedures involving vascular injury and that this event seems to occur early after vascular injury. We further conclude that CD34 affinity pheresis attenuates the surge. This approach for direct depletion of progenitors may have important implications for the study of progenitors in vascular restenosis.

First record of Molossus coibensis Allen, 1904 (Chiroptera, Molossidae) in Atlantic Forest

We report the first occurrence of the Coiban mastiff bat Molossus coibensis Allen, 1904 in the Atlantic Forest, and the third record of this species for Brazil. Our study is based on 39 adult specimens captured in Reserva Biológica de Sooretama, an Atlantic Forest preservation unit in Espírito Santo state, southeastern Brazil, apparently sharing the same shelter with M. molossus (Pallas 1776) and M. rufus E. Geoffroy, 1805. Morphological and morphometric data allows the confident separation of the two species of smallMolossus.

First record of Molossus coibensis Allen, 1904 (Chiroptera, Molossidae) in Atlantic Forest

We report the first occurrence of the Coiban mastiff bat Molossus coibensis Allen, 1904 in the Atlantic Forest, and the third record of this species for Brazil. Our study is based on 39 adult specimens captured in Reserva Biológica de Sooretama, an Atlantic Forest preservation unit in Espírito Santo state, southeastern Brazil, apparently sharing the same shelter with M. molossus (Pallas 1776) and M. rufus E. Geoffroy, 1805. Morphological and morphometric data allows the confident separation of the two species of smallMolossus.(AU)

Microbial translocation is associated with extensive immune activation in dengue virus infected patients with severe disease.

BACKGROUND: Severe dengue virus (DENV) disease is associated with extensive immune activation, characterized by a cytokine storm. Previously, elevated lipopolysaccharide (LPS) levels in dengue were found to correlate with clinical disease severity. In the present cross-sectional study we identified markers of microbial translocation and immune activation, which are associated with severe manifestations of DENV infection. METHODS: Serum samples from DENV-infected patients were collected during the outbreak in 2010 in the State of São Paulo, Brazil. Levels of LPS, lipopolysaccharide binding protein (LBP), soluble CD14 (sCD14) and IgM and IgG endotoxin core antibodies were determined by ELISA. Thirty cytokines were quantified using a multiplex luminex system. Patients were classified according to the 2009 WHO classification and the occurrence of plasma leakage/shock and hemorrhage. Moreover, a (non-supervised) cluster analysis based on the expression of the quantified cytokines was applied to identify groups of patients with similar cytokine profiles. Markers of microbial translocation were linked to groups with similar clinical disease severity and clusters with similar cytokine profiles. RESULTS: Cluster analysis indicated that LPS levels were significantly increased in patients with a profound pro-inflammatory cytokine profile. LBP and sCD14 showed significantly increased levels in patients with severe disease in the clinical classification and in patients with severe inflammation in the cluster analysis. With both the clinical classification and the cluster analysis, levels of IL-6, IL-8, sIL-2R, MCP-1, RANTES, HGF, G-CSF and EGF were associated with severe disease. CONCLUSIONS: The present study provides evidence that both microbial translocation and extensive immune activation occur during severe DENV infection and may play an important role in the pathogenesis.

Molecular systematics of the genus Artibeus (Chiroptera: Phyllostomidae).

A molecular phylogeny of the genus Artibeus using 19 of the 20 recognized species, many with samples from a broad geographic range, is presented. The analysis shows a clear distinction between the two subgenera (or genera), the 'large'Artibeus and the 'small'Dermanura, in both mitochondrial and nuclear genes. The placement and status of A. concolor remains inconclusive and is presented as the third subgenus Koopmania. The phylogenies and divergence time estimates show a marked influence of the Andes in the formation of the subgenera and the main lineages inside each subgenus. Nuclear genes showed a highly incomplete lineage sorting among species inside subgenera Artibeus and Dermanura. Indeed, shared alleles were also found between Artibeus and Koopmania, which are presumed to have split apart during the Miocene, showing that great care should be taken in using these markers. Cytochrome-b gene divergences and monophyly analyses suggest that A. lituratus and A. intermedius are indeed conspecifics. These analyses also suggested the existence of at least four 'new' species revealing a significant cryptic diversity inside the genus.

The global circulation of seasonal influenza A (H3N2) viruses.

Antigenic and genetic analysis of the hemagglutinin of approximately 13,000 human influenza A (H3N2) viruses from six continents during 2002-2007 revealed that there was continuous circulation in east and Southeast Asia (E-SE Asia) via a region-wide network of temporally overlapping epidemics and that epidemics in the temperate regions were seeded from this network each year. Seed strains generally first reached Oceania, North America, and Europe, and later South America. This evidence suggests that once A (H3N2) viruses leave E-SE Asia, they are unlikely to contribute to long-term viral evolution. If the trends observed during this period are an accurate representation of overall patterns of spread, then the antigenic characteristics of A (H3N2) viruses outside E-SE Asia may be forecast each year based on surveillance within E-SE Asia, with consequent improvements to vaccine strain selection.

Fluxo coronário desregulado durante reperfusäo pós cardioplegia / Coronary flow response is impaired during post-cardioplegia

Estudaram-se a demanda metabólica e a distribuiçäo do fluxo coronariano na presneça de fibrilaçäo ventricular (FV), durante a reperfusäo pós-cardioplegia. Foram colocados 15 suínos em circulaçäo extracorpórea e submetidos a parada cardíaca cardioplégica sangüínea anterógrada hipotérmica intermitente, durante uma hora, seguida por reperfusäo miocárdica controlada. Os animais foram divididos em três grupos (n=5), conforme estivessem em assistolia (Grupo 1) ou em FV de curta (grupo 2) ou longa duraçäo (Grupo 3), durante os dez primeiros minutos de reperfusäo. Os valores do consumo miocárdico de oxigênio (MVO2), em ml O2/min/g (média + erro padräo) durante a reperfusäo foram de 1,325 + 0,144 (grupo 1); 2,472 + 0,208 (Grupo 2) e 2,469 + 0,228 (Grupo 3). A diferença entre o MVO2 dos coraçöes em assistolia e o dos coraçöes em FV, quer de curta ou longa duraçäo, foi significante (p<0,001). A relaçäo entre os fluxos sangüíneos endo e epicárdico, bem como o fluxo sangüíneo coronário global (ml/mim/100g) foram semelhantes nos 3 grupos. Os valores dessa última variável, em ml/mim/100g, corresponderam a, respectivamente, 169,3 + 11,7; 185,0 + 15,7 e 179,9 + 13,2. Os resultados demonstram que a auto-regulaçäo coronária está alterada durante a fase inicial de reperfusäo pós criocardioplegia, pois a perfusäo miocárdica näo aumentou em resposta à elevaçäo do consumo de oxigênio imposta pela FV. Essa constataçäo, de grande interesse clínico, sugere que a ocorrência de FV durante a fase inicial da reperfusäo possa contribuir para o desenvolvimento de lesöes teciduais em coraçöes cujo fluxo coronário já esteja previamente comprometido, por obstruçäo coronária, distensäo ou hipertrofia ventricular.

Emprego de uma nova emulsäo de perfluorocarbonos (Oxygent MR) em circulaçäo extracorpórea: estudo experimental em cäes / Use of a new fluorocarbon emulsion (Oxygent MR) in extracorporeal circulation: experimental study in dogs

Testou-se uma nova emulsao de perfluorocarbonos (OxygentMR, Alliance Pharmaceutical, San Diego, CA 92121, EUA) em circulaçao extracorpórea (CEC) com hipotermia de 32 graus Celsius e hematócrito de 12 por cento. Estudaram-se 42 caes, 12 dos quais nao receberam a droga e serviram de controle (Grupo 1), enquanto os demais constituíram 3 grupos de lO animais cada, tratados com doses de Oxygent de 1,5 ml/kg (Grupo 2), 3 ml/kg (Grupo 3) e 6 ml/kg (Grupo 4) as quais geraram fluorocritos de, respectivamente, 1 por cento, 2 por cento e 3 por cento. Foram analisadas variáveis do metabolismo do oxigênio (O2) em 6 diferentes fluxos de perfusao (Q), ordenados ao acaso. Reaqueceram-se os caes, interrompeu-se a CEC e acompanharam-se os animais por l hora. Diferenças intergrupos foram analisadas pelo teste das médias dos quadrados mínimos e pelo teste de Duncan, considerando-se significantes os valores de p