Crystallization and preliminary X-ray diffraction analysis of levansucrase (LsdA) from Gluconacetobacter diazotrophicus SRT4

The endophytic bacterium Gluconacetobacter diazotrophicus SRT4 secretes a constitutively expressed levansucrase (LsdA; EC 2.4.1.10), which converts sucrose to fructo-oligosaccharides and levan. Fully active LsdA was purified to high homogeneity by non-denaturing reversed-phase HPLC and was crystallized at room temperature by the hanging-drop vapour-diffusion method using ammonium sulfate and ethanol as precipitants. The crystals are extremely sensitive, but native data have been collected to 2.5 A under cryogenic conditions using synchrotron radiation. LsdA crystals belong to the orthorhombic space group P22(1)2(1) or P2(1)2(1)2, with unit-cell parameters a = 53.80, b = 119.39, c = 215.10 A

Multifunctional albumin-stabilized gold nanoclusters for the reduction of cancer stem cells

Controlled delivery of multiple chemotherapeutics can improve the effectiveness of treatments and reduce side effects and relapses. Here in, we used albumin-stabilized gold nanoclusters modified with doxorubicin and SN38 (AuNCs-DS) as combined therapy for cancer. The chemotherapeutics are conjugated to the nanostructures using linkers that release them when exposed to different internal stimuli (Glutathione and pH). This system has shown potent antitumor activity against breast and pancreatic cancer cells. Our studies indicate that the antineoplastic activity observed may be related to the reinforced DNA damage generated by the combination of the drugs. Moreover, this system presented antineoplastic activity against mammospheres, a culturing model for cancer stem cells, leading to an efficient reduction of the number of oncospheres and their size. In summary, the nanostructures reported here are promising carriers for combination therapy against cancer and particularly to cancer stem cells.This research was funded by the Spanish Ministry of Economy and Competitiveness (CTQ2016-78454-C2-2-R, SAF2014-56763-R, and SAF2017-87305-R), Comunidad de Madrid (S2013/MIT-2850), Asociación Española Contra el Cáncer, and IMDEA Nanociencia IMDEA Nanociencia acknowledges support from the ‘Severo Ochoa’ Programme for Centres of Excellence in R&D (MINECO, Grant SEV-2016-0686

Propidium Monoazide Integrated with qPCR Enables the Detection and Enumeration of Infectious Enteric RNA and DNA Viruses in Clam and Fermented Sausages

The increase of foodborne viral outbreaks highlights the need for a rapid and sensitive method for the prediction of viral infectivity in food samples. This study assesses the use of propidium monoazide (PMA) coupled with real-time PCR methods (RT-qPCR or qPCR for RNA or DNA viruses, respectively) in the determination of viral infectivity in complex animal-related food matrices. Clam and Spanish fermented sausage (“chorizo”) samples were spiked with infectious and heat-inactivated human adenovirus-2 (HAdV-2) and mengovirus (vMC0). PMA-qPCR/RT-qPCR discriminated infective virus particles, with significant reductions (>2.7 log10 or 99.7%). Additionally, infectious HAdV-2 and vMC0 were quantified by plaque assay (in plaque forming units, PFU), and compared with those in virus genomes copies (GCs) quantified by PMA-qPCR/RT-qPCR. A consistent correlation (R2 > 0.92) was showed between PFU and GCs along serial 10-fold dilutions in both DNA and RNA virus and in both food matrices. This study shows the use of PMA coupled to qPCR/RT-qPCR as a promising alternative for prediction of viral infectivity in food samples in comparison to more expensive and time-consuming methods and for those viruses that are not able to grow under available cell culture techniques.ThisstudywasfinanciallysupportedbytheRTA2014-00024- C04-01 fromtheSpanishMinistryofEconomyandInnovation and theBrazilianCNPqProjectnumber472804/2013-8,andby CAPES/PNPD andCAPES/PDSE

CM-proteins and thionins in cereals: characterization and cloning of cDNA

The study of cereal albumins and globulins has lagged somewhat behind that of the prolamins, which nave been considered as typical reserve proteins. However, these protein fractions merit closer attention for a variety of reasons. The main individual albumins and globulins are at least as abundant as many prolamin components, and it can be speculated that in a tissue, such as the cereal endosperm, which is completely consumed during germination, al 1 abundant proteins may play a reserve role. They have also a higher proportion of essential amino acids, as compared with the prolamins, and thus may be releyant in connec^ tion with the genetic alteration of overall grain composition. Finally, a high proportion of the main components of these protein fractions have inhibitory and even toxic properties, which may be related to the protection of this tissue during development and germination, and might influence the nutritional valué of the cereal products. We report here the characterization in barley of cDNA clones encoding two major groups of proteins: the CM-proteins, a family that inciudes inhibitors of trypsin and a-amylase, and the thionins, a group of high-lysine toxic polypeptides

Increased markers of cardiac vagal activity in leucine-rich repeat kinase 2-associated Parkinson's disease.

PurposeCardiac autonomic dysfunction manifests as reduced heart rate variability (HRV) in idiopathic Parkinson's disease (PD), but no significant reduction has been found in PD patients who carry the LRRK2 mutation. Novel HRV features have not been investigated in these individuals. We aimed to assess cardiac autonomic modulation through standard and novel approaches to HRV analysis in individuals who carry the LRRK2 G2019S mutation.MethodsShort-term electrocardiograms were recorded in 14 LRRK2-associated PD patients, 25 LRRK2-non-manifesting carriers, 32 related non-carriers, 20 idiopathic PD patients, and 27 healthy controls. HRV measures were compared using regression modeling, controlling for age, sex, mean heart rate, and disease duration. Discriminant analysis highlighted the feature combination that best distinguished LRRK2-associated PD from controls.ResultsBeat-to-beat and global HRV measures were significantly increased in LRRK2-associated PD patients compared with controls (e.g., deceleration capacity of heart rate: p = 0.006) and idiopathic PD patients (e.g., 8th standardized moment of the interbeat interval distribution: p = 0.0003), respectively. LRRK2-associated PD patients also showed significantly increased irregularity of heart rate dynamics, as quantified by Rényi entropy, when compared with controls (p = 0.002) and idiopathic PD patients (p = 0.0004). Ordinal pattern statistics permitted the identification of LRRK2-associated PD individuals with 93% sensitivity and 93% specificity. Consistent results were found in a subgroup of LRRK2-non-manifesting carriers when compared with controls.ConclusionsIncreased beat-to-beat HRV in LRRK2 G2019S mutation carriers compared with controls and idiopathic PD patients may indicate augmented cardiac autonomic cholinergic activity, suggesting early impairment of central vagal feedback loops in LRRK2-associated PD

Viability of the biochar production from different manure wastes in the Amblés Valley (Ávila, Spain)

In the last years, intensive animal husbandry production has led to a large concentration of animals in small areas. This has resulted in the production of excessive amounts of manures with insufficient nearby land for application. One of this areas is the Amblés Valley located in the centre of Spain, near to Ávila city, with an extension of 167472 ha of which 88.9% is agricultural land. This valley has an important livestock focused on pig, cattle, chicken production which is associated with the generation of more than 200,000 t/year of manure. There are a number of environmental problems associated with these intensive agricultural systems, including N and P pollution of water bodies, methane emissions and odour pollution. These serious environmental threats are called for innovative environmental management approaches. A feasible technology for the management of manures, offering a potential to valorise these wastes, is pyrolysis, which results in the production of biochar. The objective of this work is evaluated the technical and economic feasibility of the production of biochar in Amblés Valley (Spain)

Evolución de una marisma y cambios del nivel del mar en el estuario de Gernika (sur del Golfo de Vizcaya)

The ecological distribution of some species of Foraminifera living in estuaries can be used in the interpretation of former coastal lines. There is a close relationship between tide level and the distribution of particular benthic foraminifer species. A characterisation of tide levels based on the benthic foraminifers in four cores from this estuary allowed us to interpret the changes over time in transitional environments during the modern Holocene. The estuary was filled during the last 6500 years following a general accretional model. Marshes have existed in this area for about 4600 years. This model is described for the first time in the area, based on benthic foraminiferal zones. These zones define a general emergence sequence, interrupted by a transgressive marine pulsation. We describe the evolution of marsh sub-environments in relation to the sea-level changes, materialised by a transgressive pulsation which shows an interruption in the accretional process dated 1910 ± 50 BP.La distribución ecológica de algunas especies de foraminíferos que viven en estuarios pueden ser utilizadas en la interpretación de antiguas líneas de costa. Existe una clara relación entre el nivel mareal y la distribución de especies concretas de foraminíferos bentónicos. La caracterización de estos niveles de mareas basados en los foraminíferos bentónicos en cuatro sondeos de este estuario, permite interpretar los cambios en estos ambientes transicionales durante el Holoceno. El estuario ha sido rellenado durante los últimos 6500 años, siguiendo un modelo general acrecional. Las marismas han existido en esta área desde hace unos 4600 años. Este modelo describe la evolución de los subambientes de marisma, en relación con cambios en el nivel del mar, materializado por una pulsación transgresiva que evidencia una interrupción en el proceso acrecional, datada de 1910 ± 50 BP