Tubular dentin sealer penetration after different final irrigation protocols: A confocal laser scanning microscopy study

The aim of this study was to evaluate tubular dentin sealer penetration, comparing different final irrigation protocols using a conventional needle (CONV), EndoActivator system (EAS), EndoVac system (EVS), and ultrasound (PUI). Initially, fifty‐two first maxillary molars with a single canal in the palatal root, without abrupt curvatures, resorptive processes, or previous endodontic treatment were selected for this study. Then, the crowns were sectioned to obtain palatal roots 15 mm in length. The root canals were prepared with the ProTaper Universal System and irrigated with 5% NaOCl. Afterwards, the specimens were divided into four groups (n. 13), according to the final irrigation protocol: CONV, EAS, EVS, and PUI. After filling, slices at 3 mm and 5 mm from the apex were obtained for analysis by confocal laser scanning microscopy. Two‐way comparisons between the groups and the levels were performed with Games Howell's test (p < .05). Tubular dentin sealer penetration was higher at 5 mm compared with 3 mm from the apex (p < .05). The EAS group showed a higher percentage of tubular dentin sealer penetration, compared with the CONV group, at both levels. At 3 mm, there was no statistically significant difference among EAS, EVS, and PUI; however, these groups showed better performance, compared with the CONV group. At 5 mm, there was no statistically significant difference between the EAS and EVS groups, but both showed higher sealer penetration than the PUI group (p < .05). The EAS and EVS groups achieved better degrees of tubular dentin sealer penetration, compared with the other groups.     

Photocuring of cycloaliphatic epoxy formulations using polyesters with multiarm star topology as additives

Multiam star polyesters were synthesized by growing poly(ε‐caprolactone) (PCL) arms from hyperbranched polyesters cores of different molecular weight and used as polymeric modifiers in UV‐curable cationic formulations based on a biscycloaliphatic epoxy resin. The effect of the multiarm stars on the curing kinetics has been investigated by real‐time FTIR. The thermal‐mechanical properties of the photocured thermosets have been studied with calorimetry and dynamomechanical and thermogravimetric analysis. Impact strength tests have been performed to assess their effect on the toughness of the cured materials. An accelerative effect of these modifiers has been observed as a consequence of the participation of the hydroxyl groups of the modifiers in the cationic curing of the epoxy resin. A modest increase in toughness accompanied by a decrease in the glass transition are observed, as a consequence of the incorporation of the modifiers into the network structure, leading to homogeneous, in situ reinforced materials. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2014 , 131, 40005.     

Seipin Deficiency as a Model of Severe Adipocyte Dysfunction: Lessons from Rodent Models and Teaching for Human Disease

Obesity prevalence is increasing worldwide, leading to cardiometabolic morbidities. Adipocyte dysfunction, impairing white adipose tissue (WAT) expandability and metabolic flexibility, is central in the development of obesity-related metabolic complications. Rare syndromes of lipodystrophy characterized by an extreme paucity of functional adipose tissue should be considered as primary adipocyte dysfunction diseases. Berardinelli-Seip congenital lipodystrophy (BSCL) is the most severe form with a near absence of WAT associated with cardiometabolic complications such as insulin resistance, liver steatosis, dyslipidemia, and cardiomyopathy. Twenty years ago, mutations in the BSCL2 gene have been identified as the cause of BSCL in human. BSCL2 encodes seipin, an endoplasmic reticulum (ER) anchored protein whose function was unknown back then. Studies of seipin knockout mice or rats demonstrated how seipin deficiency leads to severe lipodystrophy and to cardiometabolic complications. At the cellular levels, seipin is organized in multimers that are particularly enriched at ER/lipid droplet and ER/mitochondria contact sites. Seipin deficiency impairs both adipocyte differentiation and mature adipocyte maintenance. Experiments using adipose tissue transplantation in seipin knockout mice and tissue-specific deletion of seipin have provided a large body of evidence that liver steatosis, cardiomyopathy, and renal injury, classical diabetic complications, are all consequences of lipodystrophy. Rare adipocyte dysfunctions such as in BSCL are the key paradigm to unravel the pathways that control adipocyte homeostasis. The knowledge gathered through the study of these pathologies may bring new strategies to maintain and improve adipose tissue expandability.     

APOB CRISPR-Cas9 Engineering in Hypobetalipoproteinemia: A Promising Tool for Functional Studies of Novel Variants

Hypobetalipoproteinemia is characterized by LDL-cholesterol and apolipoprotein B (apoB) plasma levels below the fifth percentile for age and sex. Familial hypobetalipoproteinemia (FHBL) is mostly caused by premature termination codons in the APOB gene, a condition associated with fatty liver and steatohepatitis. Nevertheless, many families with a FHBL phenotype carry APOB missense variants of uncertain significance (VUS). We here aimed to develop a proof-of-principle experiment to assess the pathogenicity of VUS using the genome editing of human liver cells. We identified a novel heterozygous APOB-VUS (p.Leu351Arg), in a FHBL family. We generated APOB knock-out (KO) and APOB-p.Leu351Arg knock-in Huh7 cells using CRISPR-Cas9 technology and studied the APOB expression, synthesis and secretion by digital droplet PCR and ELISA quantification. The APOB expression was decreased by 70% in the heterozygous APOB-KO cells and almost abolished in the homozygous-KO cells, with a consistent decrease in apoB production and secretion. The APOB-p.Leu351Arg homozygous cells presented with a 40% decreased APOB expression and undetectable apoB levels in cellular extracts and supernatant. Thus, the p.Leu351Arg affected the apoB secretion, which led us to classify this new variant as likely pathogenic and to set up a hepatic follow-up in this family. Therefore, the functional assessment of APOB-missense variants, using gene-editing technologies, will lead to improvements in the molecular diagnosis of FHBL and the personalized follow-up of these patients.     

A comparative study of the purification of betanin

Betanin is a natural pigment with antioxidant properties used as a food colourant. This work describes the spectrophotometric and chromatographic quantification of betanin (2S/15S) and its epimer isobetanin (2S/15R) in fresh beetroot juice, food-grade beetroot powder and betanin standard diluted in dextrin. Absorption spectra of all three samples were deconvoluted using a mixed three-function model. Food-grade beetroot powder has the largest amount of violet-red impurities, probably formed during processing. The purification of betanin from these complex matrices was carried out by seven different methods. Ion exchange chromatography was the most efficient method for the purification of betanin from all samples; however, fractions contain high amounts of salt. Reversed-phase HPLC as well as reversed-phase column chromatography also produced good results at a much faster rate. The longer retention time of isobetanin when compared to betanin in reversed-phase conditions has been investigated by means of quantum-mechanical methods.     

Effects of dietary supplements of zinc-methionine on milk production,udder health and zinc metabolism in dairy goats

Twenty-two Murciano-Granadina dairy goats were used to investigate the effects of organic Zn supplementation of a diet containing a high level of inorganic Zn. Goats were kept in pens, machine milked once a day throughout lactation and fed a diet based on a dehydrated mixture of whole-plant maize and alfalfa ad libitum, alfalfa pellets, barley grain and a concentrate mixture. Treatments were: (1) control, and (2) supplemented with 1 g/d Zn-Methionine (Zn-Met) included in the concentrate mixture. After parturition, goats were blocked in week 3 and dietary treatments were applied until week 23. From weeks 3-20, feed intake, milk yield, milk composition, milk somatic cell count (SCC), and udder health were measured. In week 21, all goats were injected intraperitoneally with 1 g/d DL-methionine for 5 d to establish the effects of methionine under the conditions of udder stress induced by hand milking on the second day. During weeks 22 and 23, diet digestibility, and N and Zn balance were determined. Dry matter intake, milk yield, and milk contents of total solids, fat, total and true protein, and casein did not differ between treatments, but whey protein and non-protein nitrogen contents were significantly lower for the Zn-Met group. Milk SCC tended to decrease as a result of Zn-Met supplementation but differences between treatments were not significant when halves with persistent infection were excluded. Hand milking increased SCC in both groups, but udders of supplemented goats showed a lower reaction. Apparent absorption of N significantly increased and Zn retention tended to increase in Zn-Met supplemented goats. We conclude that Zn-Met supplementation can enhance resistance to udder stress in dairy goats. Effects were attributed to the organic Zn and not to the methionine component. Zn retention and protein utilization were also improved by the Zn-Met supplement.     

Environmental aspects of ethanol-based fuels from Brassica carinata: A case study of second generation ethanol

One of the main challenges faced by mankind in the 21st century is to meet the increasing demand for energy requirements by means of a more sustainable energy supply. In countries that are net fossil fuel importers, expectation about the benefit of using alternative fuels on reducing oil imports is the primary driving force behind efforts to promote its production and use. Spain is scarce in domestic energy sources and more than 50% of the energy used is fossil fuel based. The promotion of renewable energies use is one of the principal vectors in the Spanish energy policy. Selected herbaceous crops such as Brassica carinata are currently under study as potential energy sources. Its biomass can be considered as potential feedstock to ethanol conversion by an enzymatic process due to the characteristics of its composition, rich in cellulose and hemicellulose. This paper aims to analyse the environmental performance of two ethanol-based fuel applications (E10 and E85) in a passenger car (E10 fuel: a mixture of 10% ethanol and 90% gasoline by volume; E85 fuel: a mixture of 85% ethanol and 15% gasoline by volume) as well as their comparison with conventional gasoline as transport fuel. Two types of functional units are applied in this study: ethanol production oriented and travelling distance oriented functional units in order to reflect the availability or not of ethanol supply. E85 seems to be the best alternative when ethanol production based functional unit is considered in terms of greenhouse gas (GHG) emissions and E10 in terms of non-renewable energy resources use. Nevertheless, E85 offers the best environmental performance when travelling distance oriented functional unit is assumed in both impacts. In both functional unit perspectives, the use of ethanol-based fuels reduces the global warming and fossil fuels consumption. However, the contributions to other impact indicators (e.g. acidification, eutrophication and photochemical oxidants formation) were lower for conventional gasoline.Life Cycle Assessment (LCA) procedure helps to identify the key areas in the B. carinata ethanol production life cycle where the researchers and technicians need to work to improve the environmental performance. Technological development could help in lowering both the environmental impact and the prices of the ethanol fuels.     

Assessment of safety of enterococci isolated throughout traditional Terrincho cheesemaking: virulence factors and antibiotic susceptibility

Enterococci account for an important fraction of the adventitious microflora of traditional cheeses manufactured in Mediterranean countries from small ruminants' raw milk and play an important role in the development of suitable organoleptic characteristics of the final product. It has been suggested that animals used for food or animals that supply edible products are a reservoir of antibiotic-resistant enterococci. The main purpose of this research effort was thus to identify, to the species level, a total of 73 enterococci with high tolerance to acidic pH and bile salts (as prevailing environmental conditions in the first portion of the gastrointestinal tract), which were previously isolated from the milk feedstock to the final product of Terrincho cheesemaking, and to determine their profiles of antibiotic susceptibility, coupled with the occurrence of specific virulence factors (especially in those that might eventually be claimed to exhibit suitable probiotic and technological performances). Isolates, identified by both API 20 STREP and PCR methods, were found to belong to the following Enterococcus species: E. casseliflavus, E. durans, E. faecalis, E. faecium, and E. gallinarum. Susceptibility of those isolates was observed to most antibiotics tested, whereas none harbored aminoglycoside resistance genes. PCR screenings for cytolysin genes (cylL(L), cylL(s), cylM, cylB, and cylA), surface adhesin genes (efaA(fs), efaA(fm), and esp), the aggregation protein gene (agg), and the extracellular metalloendopeptidase gene (gelE) were performed. All isolates proved negative for cylL(L), cylM, cylB, and agg genes. Both E. faecalis strains were positive for the cell wall-associated protein Esp and the cell wall adhesin efaA(fs), whereas the cell wall adhesin efaA(fm) was detected in 11 of the 12 E. faecium strains. Only one strain possessed the cylL(s) determinant, and another possessed the cylA gene. Incidence of virulence determinants was thus very low; hence, the enterococcal adventitious microflora tested is essentially safe.     

Model‐based control addition to prescribe DFIG wind turbine fast frequency response

This paper investigates the physical capability of double‐fed induction generator (DFIG) wind turbines for inertial support of frequency response. Frequency stability is modeled using the DFIG electromechanical and generator controller dynamics, and a destabilizing effect is demonstrated in low‐inertia systems. To improve response, a synchronous reference frame DFIG controller is proposed that acts by following low‐frequency grid dynamics and adds a fast acting proportional plus integral (PI)‐controlled frequency‐responsive component to existing qd current commands. The proposed controller is derived in a straightforward manner using only the DFIG dynamic equations and is designed using pole/zero placement techniques. Laboratory experiments using a micro‐scale DFIG wind turbine with hub‐emulating flywheel prove better capability for transient frequency regulation even under extreme load change. The result is a DFIG controller that balances the appearance of transients in electrical and mechanical systems. Value is achieved in providing immediate continuous inertial response to support load change. The proposed frequency response can improve the use of existing physical inertia from wind turbines.