Humid and cold forest connections in South America between the eastern Andes and the southern Atlantic coast during the LGM.

The presence of Andean plant genera in moist forests of the Brazilian Atlantic Coast has been historically hypothesized as the result of cross-continental migrations starting at the eastern Andean flanks. Here we test hypotheses of former connections between the Atlantic and Andean forests by examining distribution patterns of selected cool and moist-adapted plant arboreal taxa present in 54 South American pollen records of the Last Glacial Maximum (LGM), ca. 19-23 cal ka, known to occur in both plant domains. Pollen taxa studied include Araucaria, Drimys, Hedyosmum, Ilex, Myrsine, Podocarpus, Symplocos, Weinmannia, Myrtaceae, Ericaceae and Arecaceae. Past connectivity patterns between these two neotropical regions as well as individual ecological niches during the LGM were explored by cluster analysis of fossil assemblages and modern plant distributions. Additionally, we examined the ecological niche of 137 plant species with shared distributions between the Andes and coastal Brazil. Our results revealed five complex connectivity patterns for South American vegetation linking Andean, Amazonian and Atlantic Forests and one disjunction distribution in southern Chile. This study also provides a better understanding of vegetation cover on the large and shallow South American continental shelf that was exposed due to a global sea level drop.

Geometry Optimization for Miniaturized Thermoelectric Generators.

Thermoelectric materials capable of converting heat into electrical energy are used in sustainable electric generators, whose efficiency has been normally increased with incorporation of new materials with high figure of merit (ZT) values. Because the performance of these thermoelectric generators (TEGs) also depends on device geometry, in this study we employ the finite element method to determine optimized geometries for highly efficient miniaturized TEGs. We investigated devices with similar fill factors but with different thermoelectric leg geometries (filled and hollow). Our results show that devices with legs of hollow geometry are more efficient than those with filled geometry for the same length and cross-sectional area of thermoelectric legs. This behavior was observed for thermoelectric leg lengths smaller than 0.1 mm, where the leg shape causes a significant difference in temperature distribution along the device. It was found that for reaching highly efficient miniaturized TEGs, one has to consider the leg geometry in addition to the thermal conductivity.

From degrader to producer: reversing the gallic acid metabolism of Pseudomonas putida KT2440.

Gallic acid is a powerful antioxidant with multiple therapeutic applications, usually obtained from the acidic hydrolysis of tannins produced by many plants. As this process generates a considerable amount of toxic waste, the use of tannases or tannase-producing microorganisms has become a greener alternative over the last years. However, their high costs still impose some barriers for industrial scalability, requiring solutions that could be both greener and cost-effective. Since Pseudomonas putida KT2440 is a powerful degrader of gallic acid, its metabolism offers pathways that can be engineered to produce it from cheap and renewable carbon sources, such as the crude glycerol generated in biodiesel units. In this study, a synthetic operon with the heterologous genes aroG4, quiC and pobA* was developed and expressed in P. putida, based on an in silico analysis of possible metabolic routes, resulting in no production. Then, the sequences pcaHG and galTAPR were deleted from the genome of this strain to avoid the degradation of gallic acid and its main intermediate, the protocatechuic acid. This mutant was transformed with the vector containing the synthetic operon and was finally able to convert glycerol into gallic acid. Production assays in shaker showed a final concentration of 346.7 ± 0.004 mg L-1 gallic acid after 72 h.

Engineering Burkholderia sacchari to enhance poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] production from xylose and hexanoate.

Burkholderia sacchari LFM101 LMG19450T is a Brazilian bacterium isolated from sugarcane crops soil and a promising biotechnological platform for bioprocesses. It is an efficient producer of poly(3-hydroxybutyrate) from carbohydrates including xylose. In the present work, the expression of B. sacchari xylose consumption genes (xylA, xylB and tktA) was combined with the expression of Aeromonas sp. phaC (PHA synthase), aiming to increase both the growth rates in xylose and the 3-hydroxyhexanoate (3HHx) molar fractions in the produced PHA. Genes were cloned into pBBR1MCS-2 vectors and then expressed in the B. sacchari PHA- mutant LFM344. Maximum specific growth rates on xylose and PHA accumulation capacity of all recombinants were evaluated. In bioreactor experiments, up to 55.5 % CDW was accumulated as copolymer, hexanoate conversion to 3HHx raised from 2 % to 54 % of the maximum theoretical value, compared to wild type. 3HHx mol% ranged from 8 to 35, and molecular weights were between 111 and 220 kg/mol. Thermal analysis measurement showed a decrease in Tg and Tm values with higher 3HHx fraction, indicating improved thermomechanical characteristics. Recombinants construction and bioreactor strategies allowed the production of P(3HB-co-3HHx) with controlled monomeric composition from xylose and hexanoate, allowing its application in diverse fields, including the medical area.

Antibiotic prophylaxis for prevention of spontaneous bacterial peritonitis in liver cirrhosis: systematic review.

BACKGROUND AND AIM: Spontaneous bacterial peritonitis is a potentially life-threatening infection in patients with liver cirrhosis and ascites. Its prevention is vital to improve prognosis of cirrhotic patients. The main objective of this systematic review was to evaluate what is the most efficacious and safest antibiotic prophylactic strategy. METHODS: Studies were located by searching PubMed and Cochrane Central Register of Controlled Trials in The Cochrane Library until February 2019. Randomized controlled trials evaluating primary or secondary spontaneous bacterial peritonitis prophylaxis in cirrhotic patients with ascites were included. The selection of studies was performed in two stages: screening of titles and abstracts, and assessment of the full papers identified as relevant, considering the inclusion criteria. Data were extracted in a standardized way and synthesized qualitatively. RESULTS: Fourteen studies were included. This systematic review demonstrated that daily norfloxacin is effective as a prophylactic antibiotic for the prevention of spontaneous bacterial peritonitis in patients with cirrhosis. Once weekly ciprofloxacin was not inferior to once daily norfloxacin, with good tolerance and no induced resistance. Trimethoprim-sulfamethoxazole and norfloxacin have similar efficacy for primary and secondary prophylaxis of spontaneous bacterial peritonitis, however, trimethoprim-sulfamethoxazole was associated with an increased risk of developing an adverse event. Rifaximin was more effective than norfloxacin in the secondary prophylaxis of spontaneous bacterial peritonitis, with a significant decrease in adverse events and mortality rate. CONCLUSIONS: Continuous long-term selective intestinal decontamination with norfloxacin is the most widely used prophylactic strategy in spontaneous bacterial peritonitis, yet other equally effective and safe options are available.

Efectividad de una intervención educativa sobre higiene de manos en unidades de hospitalización quirúrgica / Efficacy of an educational intervention on hand hygiene in Surgical Hospitalization Units

Objetivo: evaluar la eficacia de una intervención educativa sobre higiene de manos (HdM) en unidades de hospitalización quirúrgica, y su asociación con el sexo, la categoría profesional y la unidad de hospitalización de los profesionales.Método: estudio analítico cuasi-experimental pre-post de grupo único. La intervención consistió en cursos teórico-prácticos de una hora dirigidos a todos los profesionales sanitarios de la hospitalización quirúrgica del Complexo Hospitalario Universitario de Ourense, España. La unidad de observación fue las oportunidades de HdM. Se calculó un tamaño muestral de 294 oportunidades. Se realizó una comparación de proporciones de correcta HdM pre- y post-intervención empleando el test de Chi cuadrado para muestras independientes, considerándose un nivel de significación p< 0,05. Se calcularon las Razones de Prevalencia con sus intervalos de confianza al 95%.Resultados: se realizaron un total de 423 observaciones pre- y 466 post-intervención. Tras la intervención se produjo una mejora significativa de la HdM (p= 0,002), especialmente los profesionales de Enfermería (p< 0,001) y los técnicos auxiliares (p= 0,01). Los momentos que mejoraron más fueron “después de tocar el ambiente del paciente” (p= 0,001) y “antes del contacto con el paciente” (p= 0,001). Se halló un aumento significativo de esmalte en uñas (p< 0,001) y de uso de joyas, aunque esto último no significativo (p= 0,23). No se encontró ninguna diferencia significativa según sexo o unidad de hospitalización (p> 0,05).Conclusiones: una intervención formativa sobre HdM mejora significativamente la adherencia a las recomendaciones de HdM exceptuando las pautas sobre el uso de esmalte de uñas y la presencia de joyas, aunque estos datos deben ser interpretados con cautela.(AU)

Objective: to assess the efficacy of an educational intervention on hand hygiene (HH) in Surgical Hospitalization Units, and its association with the gender, professional category, and Hospitalization Unit of the professionals.Method: an analytical quasi-experimental, pre-post study, on a single arm. The intervention consisted in theoretical-practical 1-hour courses targeted to all healthcare professionals in Surgical Hospitalization at the Complexo Hospitalario Universitario de Ourense, Spain. The opportunities for HH represented the observation unit. A sample size of 294 opportunities was calculated. There was a comparison of adequate HH rates pre and post intervention, using the Chi square Test for independent samples; the level of significance considered was p.05. Prevalence Ratios were calculated with their confidence intervals at 95%.Results: in total, 423 observations were conducted pre-intervention, and 466 post-intervention. After the intervention, there was a significant improvement in HH (p= 0.002), particularly among Nursing professionals (p< 0.001) and assistant nursing technicians (p= 0.01). The instances with higher improvement were: “after touching the patient setting” (p= 0.001) and “before contact with the patient” (p= 0.001). There was a significant increase in the use of nail polish (p< 0.001) and jewelry, though the latter was non-significant (p= 0.23). No significant difference was found according to gender or Hospitalization Unit (p> 0,05).Conclusions: an educational intervention on HH significantly improved the adherence to HH recommendations, except for the guidelines regarding the use of nail polish and jewelry, although these data must be interpreted with caution.(AU)

Carriers based on poly-3-hydroxyalkanoates containing nanomagnetite to trigger hormone release.

Poly-3-hydroxybutyrate (P(3HB)) and poly-3-hydroxybutyrate-co-3-hydroxyhexanoate (P(3HB-co-3HHx)) are biocompatible and bioabsorbable biopolymers produced by different bacteria with potential for drug delivery in thermo-responsive magnetic microcarriers. Microparticles of P(3HB) and P(3HB-co-3HHx), with 5.85% mol of 3HHx, produced by Burkholderia sacchari, containing nanomagnetite (nM) and lipophilic hormone were prepared by simple emulsion (oil/water) technique leading to progesterone (Pg) encapsulation efficiency higher than 70% and magnetite loads of 3.1 and 2.3% (w/w) for P(3HB)/nM/Pg and P(3HB-co-3HHx)/nM/Pg, respectively. These formulations were characterized by Infrared spectroscopy, X-ray diffraction, Thermal gravimetric analysis and Electron microscopy (TEM, SEM) techniques. The P(3HB)/nM/Pg and P(3HB-co-3HHx)/nM/Pg microparticles presented spherical geometry with wrinkled surfaces and average size between 2 and 40 µm for 90% of the microparticles. The release profiles of the P(3HB)/nM/Pg and P(3HB-co-3HHx)/nM/Pg formulations showed a hormone release trigger (6.9 and 11.1%, respectively) effect induced by oscillating external magnetic field (0.2 T), after 72 h. Progesterone release in non-magnetic tests with P(3HB-co-3HHx)/nM/Pg revealed a slight increment (5.6%) in relation to P(3HB)/nM/Pg. The experimental release of the P(3HB)/nM/Pg and P(3HB-co-3HHx)/nM/Pg samples presented a good agreement with Higuchi model. The 3HHx comonomer content improves the hormone release of the P(3HB-co-3HHx)/nM/Pg formulation with potential for application to synchronize the estrous cycle.

Analysis of the smear layer generated by different activation systems: an in vitro study.

OBJECTIVES: To evaluate via scanning electron microscopy the amount of smear layer generated during the use of sonic and ultrasonic activation systems with standardized short-term intentional contacts with the canal walls. METHODOLOGY: Seventy single-root human teeth were randomly assigned to 2 control (n = 5 each): NC (negative control), PC (positive control) and four experimental groups (n = 15 each): NC (negative control), PC (positive control), G1 (passive ultrasonic irrigation with Irrisafe), G2 (ultrasonic irrigation with EndoUltra), G3 (sonic irrigation with Endoactivator), G4 (sonic irrigation with Eddy). Samples were instrumented with ProTaper Next®. An irrigation protocol with sodium hypochlorite (NaOCl) and ethylenediaminetetraacetic acid (EDTA) was followed for both PC and the experimental groups. Standardized intentional contacts were made in the mesial walls during 5 s at WL - 2 mm in all experimental groups. Two calibrated evaluators scored the smear layer generated with an ordinal scale by scanning electron microscopy. The weighted kappa coefficient (Kw) was calculated to determine the inter-observer agreement. Post-consensus ordinal data were analyzed using the ordinal (linear) chi-square test. RESULTS: When the agitation file is in contact with dentine walls, Irrisafe® significantly generated the least amount of smear layer in the coronal third (p < 0.05). Both in the middle and apical third, activation with Irrisafe® also showed statistically significant better results than Eddy™ and EndoUltra®. Endoactivator® showed significant better results than EndoUltra® and Eddy™ in the apical third (p < 0.05). CONCLUSIONS: Irrisafe generated the least amount of smear layer in the entire canal and similar to Endoactivator in the apical third when the agitation file is in contact with dentine walls. CLINICAL RELEVANCE: The smear layer is generated during activation. It is common to apply irrigation protocols where after the removal of the smear layer using chelants, a final rinse and activation is carried out but there are no previous studies analyzing the possible creation of a new smear layer with this final activation.

The relevance of enzyme specificity for coenzymes and the presence of 6-phosphogluconate dehydrogenase for polyhydroxyalkanoates production in the metabolism of Pseudomonas sp. LFM046.

Reconstruction of genome-based metabolic model is a useful approach for the assessment of metabolic pathways, genes and proteins involved in the environmental fitness capabilities or pathogenic potential as well as for biotechnological processes development. Pseudomonas sp. LFM046 was selected as a good polyhydroxyalkanoates (PHA) producer from carbohydrates and plant oils. Its complete genome sequence and metabolic model were obtained. Analysis revealed that the gnd gene, encoding 6-phosphogluconate dehydrogenase, is absent in Pseudomonas sp. LFM046 genome. In order to improve the knowledge about LFM046 metabolism, the coenzyme specificities of different enzymes was evaluated. Furthermore, the heterologous expression of gnd genes from Pseudomonas putida KT2440 (NAD+ dependent) and Escherichia coli MG1655 (NADP+ dependent) in LFM046 was carried out and provoke a delay on cell growth and a reduction in PHA yield, respectively. The results indicate that the adjustment in cyclic Entner-Doudoroff pathway may be an interesting strategy for it and other bacteria to simultaneously meet divergent cell needs during cultivation phases of growth and PHA production.

Double-Microcatheter Technique through Tortuous Anatomy for Coil Embolization of a Saccular $plenic Aneurysm: a Technical Report.

We report on a case of an asymptomatic splenic artery aneurysm (SAA) with a large neck in a 53-year-old female with an extreme vessel tortuosity which was treated with a Double Microcatheter Technique. This endovascular procedure consists of embolization of the aneurysm using detachable coils with no application of any glue, stent or balloon. At the end of procedure, no complications occurred. At the three-month follow-up an MRI showed the aneurysm's complete exclusion and patency of the splenic artery.

Brazilian montane rainforest expansion induced by Heinrich Stadial 1 event.

The origin of modern disjunct plant distributions in the Brazilian Highlands with strong floristic affinities to distant montane rainforests of isolated mountaintops in the northeast and northern Amazonia and the Guyana Shield remains unknown. We tested the hypothesis that these unexplained biogeographical patterns reflect former ecosystem rearrangements sustained by widespread plant migrations possibly due to climatic patterns that are very dissimilar from present-day conditions. To address this issue, we mapped the presence of the montane arboreal taxa Araucaria, Podocarpus, Drimys, Hedyosmum, Ilex, Myrsine, Symplocos, and Weinmannia, and cool-adapted plants in the families Myrtaceae, Ericaceae, and Arecaceae (palms) in 29 palynological records during Heinrich Stadial 1 Event, encompassing a latitudinal range of 30°S to 0°S. In addition, Principal Component Analysis and Species Distribution Modelling were used to represent past and modern habitat suitability for Podocarpus and Araucaria. The data reveals two long-distance patterns of plant migration connecting south/southeast to northeastern Brazil and Amazonia with a third short route extending from one of them. Their paleofloristic compositions suggest a climatic scenario of abundant rainfall and relative lower continental surface temperatures, possibly intensified by the effects of polar air incursions forming cold fronts into the Brazilian Highlands. Although these taxa are sensitive to changes in temperature, the combined pollen and speleothems proxy data indicate that this montane rainforest expansion during Heinrich Stadial 1 Event was triggered mainly by a less seasonal rainfall regime from the subtropics to the equatorial region.

Investigating Nutrient Limitation Role on Improvement of Growth and Poly(3-Hydroxybutyrate) Accumulation by Burkholderia sacchari LMG 19450 From Xylose as the Sole Carbon Source.

Burkholderia sacchari LMG19450, a non-model organism and a promising microbial platform, was studied to determine nutrient limitation impact on poly(3-hydroxybutyrate) [P(3HB)] production and bacterial growth from xylose, a major hemicellulosic residue. Nitrogen and phosphorus limitations have been studied in a number of cases to enhance PHA accumulation, but not combining xylose and B. sacchari. Within this strategy, it was sought to understand how to control PHA production and even modulate monomer composition. Nitrogen-limited and phosphorus-limited fed-batch experiments in bioreactors were performed to evaluate each one's influence on cell growth and poly(3-hydroxybutyrate) production. The mineral medium composition was defined based on yields calculated from typical results so that nitrogen was available during phosphorus limitation and residual phosphorus was available when limiting nitrogen. Sets of experiments were performed so as to promote cell growth in the first stage (supplied with initial xylose 15 g/L), followed by an accumulation phase, where N or P was the limiting nutrient when xylose was fed in pulses to avoid concentrations lower than 5 g/L. N-limited fed-batch specific cell growth (around 0.19 1/h) and substrate consumption (around 0.24 1/h) rates were higher when compared to phosphorus-limited ones. Xylose to PHA yield was similar in both conditions [0.37 gP(3HB)/gxyl]. We also described pst gene cluster in B. sacchari, responsible for high-affinity phosphate uptake. Obtained phosphorus to biomass yields might evidence polyphosphate accumulation. Results were compared with studies with B. sacchari and other PHA-producing microorganisms. Since it is the first report of the mentioned kinetic parameters for LMG 19450 growing on xylose solely, our results open exciting perspectives to develop an efficient bioprocess strategy with increased P(3HB) production from xylose or xylose-rich substrates.

Biomimetic Electronic Devices for Measuring Bacterial Membrane Disruption.

Antibiotic discovery has experienced a severe slowdown in terms of discovery of new candidates. In vitro screening methods using phospholipids to model the bacterial membrane provide a route to identify molecules that specifically disrupt bacterial membranes causing cell death. Thanks to the electrically insulating properties of the major component of the cell membrane, phospholipids, electronic devices are highly suitable transducers of membrane disruption. The organic electrochemical transistor (OECT) is a highly sensitive ion-to-electron converter. Here, the OECT is used as a transducer of the permeability of a lipid monolayer (ML) at a liquid:liquid interface, designed to read out changes in ion flux caused by compounds that interact with, and disrupt, lipid assembly. This concept is illustrated using the well-documented antibiotic Polymixin B and the highly sensitive quantitation of permeability of the lipid ML induced by two novel recently described antibacterial amine-based oligothioetheramides is shown, highlighting molecular scale differences in their disruption capabilities. It is anticipated that this platform has the potential to play a role in front-line antimicrobial compound design and characterization thanks to the compatibility of semiconductor microfabrication technology with high-throughput readouts.

Engineering xylose metabolism for production of polyhydroxybutyrate in the non-model bacterium Burkholderia sacchari.

BACKGROUND: Despite its ability to grow and produce high-value molecules using renewable carbon sources, two main factors must be improved to use Burkholderia sacchari as a chassis for bioproduction at an industrial scale: first, the lack of molecular tools to engineer this organism and second, the inherently slow growth rate and poly-3-hydroxybutyrate [P(3HB)] production using xylose. In this work, we have addressed both factors. RESULTS: First, we adapted a set of BglBrick plasmids and showed tunable expression in B. sacchari. Finally, we assessed growth rate and P(3HB) production through overexpression of xylose transporters, catabolic or regulatory genes. Overexpression of xylR significantly improved growth rate (55.5% improvement), polymer yield (77.27% improvement), and resulted in 71% of cell dry weight as P(3HB). CONCLUSIONS: These values are unprecedented for P(3HB) accumulation using xylose as a sole carbon source and highlight the importance of precise expression control for improving utilization of hemicellulosic sugars in B. sacchari.