Type 2 Diabetes and all-cause mortality among Spanish women with breast cancer.

PURPOSE: To explore the effect of type 2 diabetes mellitus (T2DM) on the risk of death among women with breast cancer (BC). METHODS: A survival analysis was conducted among a cohort of women diagnosed with BC between 2006 and 2012 in Spain (n = 4,493). Biopsy or surgery confirmed BC cases were identified through the state population-based cancer registry with information on patients' characteristics and vital status. Physician-diagnosed T2DM was confirmed based on primary health care clinical history. Cox regression analyses were used to estimate adjusted hazard ratios (aHR) and 95% confidence intervals (CI) for all-cause death. Analyses were adjusted for age, hospital size, several clinical characteristics (including BC stage and histology, among others) and treatment modalities. RESULTS: Among the 4,493 BC women, 388 (8.6%) had coexisting T2DM. Overall, 1,299 (28.9%) BC women died during the completion of the follow-up and 785 (17.5%) did so during the first five years after BC diagnosis, resulting in a five-year survival rate of 82.5%. The death rate was higher in women with T2DM (43.8% died during whole period and 26.0% during the first five years) when compared with women without T2DM (27.5% and 16.7%, respectively). Accordingly, all-cause mortality was higher in women with T2DM (aHR: 1.22; 95% CI 1.03-1.44), especially if T2DM was diagnosed before BC (aHR:1.24; 95% CI 1.03-1.50) and in women with BC diagnosed before 50 years (aHR: 2.38; 95% CI 1.04-5.48). CONCLUSIONS: T2DM was associated with higher all-cause mortality among Spanish women with BC, particularly when the T2DM diagnosis was prior to the BC.

Evaluation of hippuric acid content in goat milk as a marker of feeding regimen.

Organic producers, traders, and consumers must address 2 issues related to milk: authentication of the production system and nutritional differentiation. The presence of hippuric acid (HA) in goat milk samples has been proposed as a possible marker to differentiate the feeding regimen of goats. The objective of this work is to check the hypothesis that HA could be a marker for the type of feeding regimen of goats by studying the influence of production system (conventional or organic) and feeding regimen (with or without grazing fodder). With this purpose, commercial cow and goat milk samples (n=27) and raw goat milk samples (n=185; collected from different breeds, localizations, and dates) were analyzed. Samples were grouped according to breed, feeding regimen, production system, and origin to compare HA content by ANOVA and honestly significant difference Tukey test at a confidence level of ≥95%. Hippuric acid content was obtained by analyzing milk samples with capillary electrophoresis. This method was validated by analyzing part of the samples with HPLC as a reference technique. Sixty-nine raw goat milk samples (of the total 158 samples analyzed in this work) were quantified by capillary electrophoresis. In these samples, the lowest average content for HA was 7±3 mg/L. This value corresponds to a group of conventional raw milk samples from goats fed with compound feed. The highest value of this group was 28±10 mg/L, corresponding to goats fed compound feed plus grass. Conversely, for organic raw goat milk samples, the highest concentration was 67±14 mg/L, which corresponds to goats fed grass. By contrast, the lowest value of this organic group was 26±10 mg/L, which belongs to goats fed organic compounds. Notice that the highest HA average content was found in samples from grazing animals corresponding to the organic group. This result suggests that HA is a good marker to determine the type of goats feeding regimen; a high content of HA represents a diet based mainly or exclusively on eating green grass (grazing), independently of the production system. Hence, this marker would not be useful for the actual organic policies to distinguish organic milk under the current regulations, because organic dairy ruminants can be fed organic compound feed and conserved fodder without grazing at all.

Analytical Tools for Disease Diagnosis in Animals via Fecal Volatilome.

Volatilome analysis is growing in attention for the diagnosis of diseases in animals and humans. In particular, volatilome analysis in fecal samples is starting to be proposed as a fast, easy and noninvasive method for disease diagnosis. Volatilome comprises volatile organic compounds (VOCs), which are produced during both physiological and patho-physiological processes. Thus, VOCs from a pathological condition often differ from those of a healthy state and therefore the VOCs profile can be used in the detection of some diseases. Due to their strengths and advantages, feces are currently being used to obtain information related to health status in animals. However, they are complex samples, that can present problems for some analytical techniques and require special consideration in their use and preparation before analysis. This situation demands an effort to clarify which analytic options are currently being used in the research context to analyze the possibilities these offer, with the final objectives of contributing to develop a standardized methodology and to exploit feces potential as a diagnostic matrix. The current work reviews the studies focused on the diagnosis of animal diseases through fecal volatilome in order to evaluate the analytical methods used and their advantages and limitations. The alternatives found in the literature for sampling, storage, sample pretreatment, measurement and data treatment have been summarized, considering all the steps involved in the analytical process.

A non-destructive sampling method for food authentication using gas chromatography coupled to mass spectrometry or ion mobility spectrometry.

The study of volatile compounds obtained by gas chromatography (GC) coupled to mass spectrometry (MS) or ion mobility spectrometry (IMS) may be very useful to protect food quality, especially when using a non-destructive sampling method. In this work, the authentication of the highly appreciated dry-cured Iberian ham by those techniques was studied and compared. The results obtained show the suitability of a non-destructive sampling method coupled to headspace sampling (HS)-GC-IMS or HS-GC-MS to determine volatile markers in the feeding Iberian pig regime. Although both methods were suitable to differentiate the ham categories, HS-GC-IMS was more sensitive detecting a higher number of compounds than HS-GC-MS, which provided accurate qualitative results. The results of principal component analysis showed that ethanol, 2-propanol and 3-methylbutanol, identified by HS-GC-IMS, and 3-methylbutanal and heptane, identified by HS-GC-MS, could be considered potential markers to identify ham from different feeding regimes.

Oral Vaccination with Hepatitis E Virus Capsid Protein and Immunobiotic Bacterium-Like Particles Induce Intestinal and Systemic Immunity in Mice.

The hepatitis E virus (HEV) genotype 3 (GT3) is an emergent pathogen in industrialized countries. It is transmitted zoonotically and may lead to chronic hepatitis in immunocompromised individuals. We evaluated if the major antigen of HEV, the capsid protein, can be used in combination with immunobiotic bacterium-like particles (IBLP) for oral vaccination in a mouse model. We have cloned and expressed the RGS-His5-tagged HEV GT3 capsid protein (ORF2) in E. coli and purified it by NiNTA. IBLP were obtained from two immunobiotic Lactobacillus rhamnosus strains acid- and heat-treated. ORF2 and the IBLP were orally administered to Balb/c mice. After three oral immunizations (14-day intervals), blood, intestinal fluid, Peyer´s patches, and spleen samples were drawn. IgA- and IgG-specific antibodies were determined by ELISA. Mononuclear cell populations from Peyer's patches and spleen were analyzed by flow cytometry, and the cytokine profiles were determined by ELISA to study cellular immunity. Orally administered recombinant ORF2 and IBLP from two L. rhamnosus strains (CRL1505 and IBL027) induced both antigen-specific humoral and cellular immune responses in mice. IBLP027 was more effective in inducing specific secretory IgA in the gut. IFN-γ, TNF-α, and IL-4 were produced by Peyer's plaques lymphocytes stimulated with ORF2 ex vivo suggesting a mixed Th1/Th2-type adaptive immune response in immunized mice. Oral vaccines are not invasive, do not need to be administered by specialized personal, and elicit both systemic and local immune responses at the port of entry. Here, we present an experimental oral vaccine for HEV GT3, which could be further developed for human and/or veterinary use.

Occurrence of TRGV-BJ hybrid gene in SV40-transformed fibroblast cell lines.

Illegitimate V(D)J-recombination in lymphoid malignancies involves rearrangements in immunoglobulin or T-cell receptor genes, and these rearrangements may play a role in oncogenic events. High frequencies of TRGV-BJ hybrid gene (rearrangement between the TRB and TRG loci at 7q35 and 7p14-15, respectively) have been detected in lymphocytes from patients with ataxia telangiectasia (AT), and also in patients with lymphoid malignancies. Although the TRGV-BJ gene has been described only in T-lymphocytes, we previously detected the presence of TRGV-BJ hybrid gene in the genomic DNA extracted from SV40-transformed AT5BIVA fibroblasts from an AT patient. Aiming to determine whether the AT phenotype or the SV40 transformation could be responsible for the production of the hybrid gene by illegitimate V(D)J-recombination, DNA samples were extracted from primary and SV40-transformed (normal and AT) cell lines, following Nested-PCR with TRGV- and TRBJ-specific primers. The hybrid gene was only detected in SV40-transformed fibroblasts (AT-5BIVA and MRC-5). Sequence alignment of the cloned PCR products using the BLAST program confirmed that the fragments corresponded to the TRGV-BJ hybrid gene. The present results indicate that the rearrangement can be produced in nonlymphoid cells, probably as a consequence of the genomic instability caused by the SV40-transformation, and independently of ATM gene mutation.

Comparison of off- and in-line solid-phase extraction for enhancing sensitivity in capillary electrophoresis using ochratoxin as a model compound.

This paper proposes and compares two approaches based on off- and in-line solid-phase extraction (SPE), intended to enhance sensitivity in capillary electrophoresis with ultraviolet detection (CE-UV) using as a model the determination of ochratoxin A (OA) in river water samples. In the off-line SPE mode, the reversed-phase sorbent (octadecilsylane, C(18)) selectively retains the target analyte (OA) and allows large volumes of the sample (70 mL) to be introduced and subsequently eluted in a small volume (0.1 mL) of an appropriate solution. In the in-line SPE mode, a custom-made microcartridge is inserted near the inlet of the capillary, which is filled with the same C(18) sorbent. This solid phase selectively retains OA present in a sample volume as low as approximately 640 microL for subsequent elution with ca. 135 nL of an appropriate eluent. The limit of detection (LOD) obtained with the in-line SPE method was 1 ng L(-1), which is 3 orders of magnitude lower than that obtained with CE-UV and roughly 1 order lower than that provided by the off-line SPE-CE-UV method.

Corrigendum to "Identifying cloud forest conservation areas in Mexico from the potential distribution of 19 representative species" [Heliyon 5 (3) (March 2019) e01423].

[This corrects the article DOI: 10.1016/j.heliyon.2019.e01423.].

Identifying cloud forest conservation areas in Mexico from the potential distribution of 19 representative species.

Cloud forest is a sensitive and vulnerable ecosystem that is threatened by human activities as well as climate change. Previous studies have shown how transitional ecosystems such as cloud forests will be the most negatively impacted by the global increase in temperature. Therefore, the niche modeling framework was used in this study to geographically identify the areas with the climatic potential to host the largest number of key tree species in this ecosystem and to propose them as priority conservation areas. A total of 19 species were modeled using the MaxEnt algorithm; binary maps were generated for each species and combined to produce one potential suitability map and identify climatic priority areas. Thus, 7% of the national area of Mexico shows suitability for the cloud forest ecosystem, although it is currently distributed in less than 1% of the country. Finally, potential suitability areas were compared with natural protected areas, current land use and priority conservation areas. We found that of the current suitable area, only 5% coincides with some federal or state protection regime. Natural protected areas have proven to be a mechanism for forest conservation, so we must consider increasing the number and area of those protected areas that favor the conservation of these key cloud forest species.

Direct determination of volatile analytes from solid samples by UV-ion mobility spectrometry.

A simple, reliable, inexpensive sample introduction system (SIS) coupled to an ion mobility spectrometry (IMS) equipment was used for the determination of volatile analytes present in solid samples. Following minimal pretreatment, the solid sample (e.g. fish) is directly placed in a membrane unit and released analytes (trimethylamine, dimethylamine and ammonia) pass through the membrane into a nitrogen acceptor stream; finally, passing the gaseous stream trough the UV-IMS system allows the analytes to be readily determined. Volatiles are released by heating the sample at 75 degrees C for 5 min following addition of a few drops of NaOH solution. This method is quite expeditious and uses small amounts of sample. The calibration graph is linear over the concentration range 5-225 microg mL(-1). The proposed SIS-IMS method is quite repeatable (RSD<4%) and reproducible (RSD<9%). Also, it provides acceptable analyte recoveries (111+/-17%) from spiked fish samples. The results of this study testify to the potential of IMS for qualitative and quantitative analytical determinations.

Propuesta metodológica para el uso del programa PCXMC 2.0 en modalidad rotacional / Methodological Proposal for the use of the program PCXMC 2.0 in Rotational Mode

RESUMEN Durante los procedimientos intervencionistas que utilizan fluoroscopia, se entrega a los pacientes diferentes niveles de dosis de radiación, que pueden tener un impacto a corto, mediano o largo plazo, así como en su severidad. Esto en función del tipo de radiación utilizada y de la sensibilidad a la radiación del tipo(s) de órgano(s) irradiado, es en este punto que se vuelve determinante conocer la dosis a órganos entregada durante los procedimientos con el fin de asegurar el bienestar de los pacientes, este procedimiento no se puede realizar directamente en los órganos del paciente, por lo que se utilizan programas especializado en cálculos dosimétricos. El software PCXMC 2.0 logra estimar la dosis efectiva a través del método determinístico Monte Carlo, a su vez, agregando la dosis equivalente y evaluación de riesgo para estudios dosimétricos, por lo cual, el objetivo del presente trabajo fue proponer una metodología estandarizada para la utilización del programa PCXMC en el cálculo de estas dos magnitudes

ABSTRACT During interventional procedures that utilize fluoroscopy, patients are exposed to varying levels of radiation doses, which can have short, medium, or long term impacts, as well as varying severities. This depends on the type of radiation used and the radiation sensitivity of the organ(s) being irradiated. At this point, it becomes crucial to determine the organ doses delivered during procedures in order to ensure patient well-being. Since direct measurements cannot be made on the patient's organs, specialized software is used for dosimetric calculations. PCXMC 2.0 software achieves the estimation of effective dose through the deterministic Monte Carlo method, while also incorporating equivalent dose and risk assessment for dosimetric studies. Therefore, the objective of this work was to propose a standardized methodology for the use of the PCXMC 2.0 sofware in the calculation of these two magnitudes.it is important to outline a working method for this software and provide instructions on its operation

Diversity of LEF/TCF action in development and disease.

Lymphoid enhancer factor/T cell factor proteins (LEF/TCFs) mediate Wnt signals in the nucleus by recruiting beta-catenin and its co-activators to Wnt response elements (WREs) of target genes. This activity is important during development but its misregulation plays a role in disease such as cancer, where overactive Wnt signaling drives LEF/TCFs to transform cells. The size of the LEF/TCF family is small: approximately four members in vertebrates and one orthologous form in flies, worms and hydra. However, size belies complexity. The LEF/TCF family exhibits extensive patterns of alternative splicing, alternative promoter usage and activities of repression, as well as activation. Recent work from numerous laboratories has highlighted how this complexity has important biological consequences in development and disease.

Self-assembled monolayer-based piezoelectric flow immunosensor for the determination of canine immunoglobulin.

A simple, highly sensitive immunosensor for the direct determination of immunoglobulin (Ig) in canine serum based on a piezoelectric crystal accommodated in a flow-cell was developed and optimized. The new biosensor is also useful for discriminating between Ig subclasses present in canine serum by using specific monoclonal antibodies binding to the coated crystal. Various canine monoclonal anti-IgG were deposited onto the surface of the gold-coated crystal resonator using the self-assembly technique to form a receptor layer. The highly ordered self-assembled monolayers thus obtained provide a well-controlled surface structure and many advantages as regards sensing performance. The results obtained with the proposed immunosensor were compared with those provided by a protein A-based orientation-controlled immobilization method for the same monoclonal antibodies and also with direct physical adsorption of the antibodies. The crystal was accommodated in a flow-cell that was inserted into a buffer flowing stream in order to make resonant frequency measurements.

Membrane set up combined with photoionization-ion mobility spectrometer to improve analytical performance and avoid humidity interference on the determination of aromatics in gaseous samples.

UV-ion mobility spectrometry (UV-IMS) is a reliable and inexpensive technique which allows efficient monitoring of BTX (benzene, toluene, m-xylene, o-xylene and p-xylene) in different air samples. Water molecules are unavoidably present in every on-field measurement affecting sensitivity and selectivity of the UV-IMS method. For this reason, the influence of humidity on the mobility spectra when measuring BTX is discussed here. Furthermore, a polydimethyl siloxane (PDMS) membrane assembled on an ad-hoc designed membrane holder coupled to UV-IMS was proposed for online measurement of analytes in humid gaseous samples without sample preparation/pre-treatment steps. The use of this membrane reduces the moisture of the gaseous sample before entering into the IMS, and the consequent distortion of the signal. The limit of detection (LOD) and the limit of quantification (LOQ) achieved with the method proposed were in the range of 0.49-1.21mgL(-1) and 1.63-4.03mgL(-1), respectively for all analytes. The precision of the method was evaluated in terms of repeatability and reproducibility obtaining values lower than 1.1% for drift time and 12.0% for peak height when the membrane was used coupled to the UV-IMS for all target analytes in the humidity range of 10-75% RH. Thus, BTX can be determined directly and quantified unequivocally with the membrane system in ambient air even at humid condition.

Development of a QuEChERS-based extraction method for the determination of destruxins in potato plants by UHPLC-MS/MS.

Ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) with electrospray ionization has been proposed for the determination of fifteen natural destruxins (A, B, C, D, E, Ed, Ed1, A2, B2, D2, E2, Cl, DesmA, DesmB, and DH-A), secondary metabolites with insecticidal and phytotoxic activities produced by Metarhizium species fungus, which are being studied as biological agents in pest control. Therefore, procedures to control them in the food chain are required, starting with crops. As a consequence, in this study, a simple QuEChERS-based destruxin (dtx) extraction procedure has been developed and validated in four different parts of potato plant (tuber, root, stem and leaves) for the first time. For dtx A, the limits of detection obtained, ranged between 0.5 and 1.3 µg/kg, and for quantification, ranged between 1.7 and 4.2 µg/kg. Precision values were below 8.5%; and in all cases, recoveries were higher than 91%. Finally, the method has been applied in potato samples inoculated by EAMa 01/58-Su strain, where dtxs A and B were detected and quantified. In all cases, dtx B concentration was higher than dtx A.

Simultaneous determination of benzene and phenol in heat transfer fluid by head-space gas chromatography hyphenated with ion mobility spectrometry.

The quantitative determination of some compounds such as benzene and phenol in a complex matrix by ion mobility spectrometry (IMS) can be a difficult task, due to the influence of other components present in the matrix and the chemical properties of both compounds, such as their high volatility and low proton affinity. Monitoring of these compounds in a heat transfer fluid (HTF) is essential to check the correct working of a thermosolar plant and for safety and environmental reasons. Benzene and phenol, among other compounds, are produced when HTF is exposed to high temperatures in continuous cycles and their presence can decrease the efficiency of HTF. For the first time, a headspace module coupled to a gas chromatography column in combination with an IMS (with a tritium ionization source) has been optimized and fully validated to simultaneously quantify benzene and phenol in HTF. The limit of detection (LOD) and limit of quantification (LOQ) achieved with the method proposed were 0.011 and 0.038 g L(-1) and 0.004 and 0.014 g L(-1) for benzene and phenol respectively. The precision of the method was evaluated in terms of repeatability and reproducibility with all values lower than 9.2% and 13.3%, respectively. Results demonstrated that benzene and phenol were generated in the HTF heating process, and its concentration increased with heating time (approximately 483 h). The average concentration values for benzene and phenol in degraded HTF samples were not significantly different to values obtained using a gas chromatography-flame ionization detector instrument. Therefore, IMS is a promising technique for in-field quality control of HTF in a thermosolar plant due to its speed, versatility, sensitivity and selectivity to quantify these degradation compounds.

Gene expression array of HTLV type 1-infected T cells: Up-regulation of transcription factors and cell cycle genes.

By utilizing a human cDNA expression array blot (588 genes), we have observed overexpression of various transcription factors, cell cycle regulated kinases, and DNA repair genes in HTLV-1-infected T cells. One of the genes of interest, and focus in this study, is the cyclin-dependent kinase inhibitor, p21/waf1. The p21/waf1 transcription and protein is overexpressed in all HTLV-1-infected cell lines tested as well as ATL and HAM/TSP patient samples. While p21/waf1 has been shown to display a selectivity for G(1)/S cyclin/cdk complexes, we have observed p21/waf1 to be complexed with cyclin A/cdk2. Functionally, the association of p21/cyclin A/cdk2 decreased the histone H1 phosphorylation in vitro, as observed in immunoprecipitations followed by kinase assays, as well as affecting other substrates such as the C-terminus of Rb protein involved in c-Abl and HDAC1 regulation. Wild-type, but not a mutant form (M47) of Tax, was found to be able to transactivate the p21/waf1 promoter in a p53-independent manner. We found that the minimal p21/waf1 promoter (-49 to +49 sequence) was activated by Tax and the minimal promoter contained two E2A transcription factor binding sites located between the TATA box and the initiation site. E2A proteins, E12 and E47, as well as a related helix-loop-helix protein, HEB, are all up-regulated in HTLV-1-infected T cells. When using band shift analysis, we found that only the E1 site (overlapping the transcription start site) was a functional DNA binding site. By using a chromatin immunoprecipitation (ChIP) assay, we observed that histone H4, and not histone H3, was acetylated from the endogenous p21/waf1 promoter in vivo, implying that CBP/p300, and not the SAGA complex, was critical in complexing with E2A in up-regulation of p21/waf1 in HTLV-1-infected cells.

Coupling continuous separation techniques to capillary electrophoresis.

One of the weak points of capillary electrophoresis is the need to implement rigorously sample pretreatment because its great impact on the quality of the qualitative and quantitative results provided. One of the approaches to solve this problem is through the symbiosis of automatic continuous flow systems (CFSs) and capillary electrophoresis (CE). In this review a systematic approach to CFS-CE coupling is presented and discussed. The design of the corresponding interface depends on three factors, namely: (a) the characteristics of the CFS involved which can be non-chromatographic and chromatographic; (b) the type of CE equipment: laboratory-made or commercially available; and (c) the type of connection which can be in-line (on-capillary), on-line or mixed off/on-line. These are the basic criteria to qualify the hyphenation of CFS (solid-phase extraction, dialysis, gas diffusion, evaporation, direct leaching) with CE described so far and applied to determine a variety of analytes in many different types of samples. A critical discussion allows one to demonstrate that this symbiosis is an important topic in research and development, besides separation and detection, to consolidate CE as a routine analytical tool.

Direct determination of biogenic amines in wine by integrating continuous flow clean-up and capillary electrophoresis with indirect UV detection.

A flow-injection manifold for automating the determination of biogenic amines in wine using capillary electrophoresis (CE) with indirect UV detection was developed. The ensuing method involves clean-up and solid-phase extraction (SPE) of the target analytes in the sample. Various treatments involving different SPE minicolumns were tested and compared. The C18 minicolumn was chosen to concentrate the amines following addition of ammonium chloride and ammonium hydroxide as buffer to neutralize them. Additions of amine standards were used to determine recoveries. Biogenic amines can be separated and detected after SPE with limits of detection in the range 0.05-0.1 microgram ml-1 by using 4 mM copper(II) sulphate, formic acid and 18-crown-6 as running buffer. All the amines studied are eluted within 15 min under the optimum conditions established. The overall process was successfully used to identify biogenic amines in various types of wine from different Spanish regions.

Determination of anti-carcinogenic polyphenols present in green tea using capillary electrophoresis coupled to a flow injection system.

A capillary electrophoresis (CE) method was developed for the simultaneous determination of a number of major ingredients of green tea. The components analysed were caffeine, adenine, theophylline, epigallocatechin-3 gallate, epigallocatechin, epicatechin-3 gallate, (-)-epicatechin, (+)-catechin, gallic acid, quercetin and caffeic acid. Separation was achieved using a fused capillary column with 0.15 M H3BO3 as buffer at a pH of 8.5, UV detection at 210 nm and 20 kV of voltage. Analysis was carried out after treatment (extraction, filtration and dilution) of the samples in a flow injection system which was coupled to a CE equipment via a programmable arm. The procedure allows the determination of these compounds in less than 20 min. Quantitative analysis was performed by the standard addition method. Limits of detection ranged between 0.04 microgram ml-1 for flavonols and 1.2 micrograms ml-1 for caffeine.