223 research outputs found
Impact of Extracellular Vesicle Isolation Methods on Downstream miRNA Analysis in Semen: A Comparative Study
Seminal plasma (SP) contains a unique concentration of miRNA, mostly contained in small extracellular vesicles (sEVs) such as exosomes, some of which could be clinically useful for diagnosis and/or prognosis of urogenital diseases such as prostate cancer (PCa). We optimized several exosome-EV isolation technologies for their use in semen, evaluating EV purifying effectiveness and impact on the downstream analysis of miRNAs against results from the standard ultracentrifugation (UC) method to implement the use of SP sEV_miRNAs as noninvasive biomarkers for PCa. Our results evidenced that commercial kits designed to isolate exosomes/EVs from blood or urine are mostly applicable to SP, but showed quantitative and qualitative variability between them. ExoGAG 3500× g and the miRCURY Cell/Urine/CSF 1500× g methods resulted as equivalent alternative procedures to UC for isolating exosomes/sEVs from semen for nanoparticle characteristics and quality of RNA contained in vesicles. Additionally, the expression profile of the altered semen sEV-miRNAs in PCa varies depending on the EV isolation method applied. This is possibly due to different extraction techniques yielding different proportions of sEV subtypes. This is evidence that the exosome-EV isolation method has a significant impact on the analysis of the miRNAs contained within, with important consequences for their use as clinical biomarkers. Therefore, miRNA analysis results for EVs cannot be directly extrapolated between different EV isolation methods until clear markers for delineation between microvesicles and exosomes are established. However, EV extraction methodology affects combined models (semen exosome miRNA signatures plus blood Prostate specific antigen (PSA) concentration for PCa diagnosis) less; specifically our previously described (miR-142-3p + miR-142-5p + miR-223-3p + PSA) model functions as molecular marker from EVs from any of the three isolation methods, potentially improving the efficiency of PSA PCa diagnosis
3′IsomiR Species Composition Affects Reliable Quantification of miRNA/isomiR Variants by Poly(A) RT-qPCR: Impact on Small RNA-Seq Profiling Validation
Small RNA-sequencing (small RNA-seq) has revealed the presence of small RNA-naturally occurring variants such as microRNA (miRNA) isoforms or isomiRs. Due to their small size and the sequence similarity among miRNA isoforms, their validation by RT-qPCR is challenging. We previously identified two miR-31-5p isomiRs-the canonical and a 3 ' isomiR variant (3 ' G addition)-which were differentially expressed between individuals with azoospermia of different origin. Here, we sought to determine the discriminatory capacity between these two closely-related miRNA isoforms of three alternative poly(A) based-RT-qPCR strategies in both synthetic and real biological context. We found that these poly(A) RT-qPCR strategies exhibit a significant cross-reactivity between these miR-31-5p isomiRs which differ by a single nucleotide, compromising the reliable quantification of individual miRNA isoforms. Fortunately, in the biological context, given that the two miRNA variants show changes in the same direction, RT-qPCR results were consistent with the findings of small RNA-seq study. We suggest that miRNA selection for RT-qPCR validation should be performed with care, prioritizing those canonical miRNAs that, in small RNA-seq, show parallel/homogeneous expression behavior with their most prevalent isomiRs, to avoid confounding RT-qPCR-based results. This is suggested as the current best strategy for robust biomarker selection to develop clinically useful tests
Long-Term Care Facilities and Nursing Homes during the First Wave of the COVID-19 Pandemic: A Scoping Review of the Perspectives of Professionals, Families and Residents
The COVID-19 pandemic has had significant repercussions for nursing home residents, their families, and professionals. The objective was to describe the perspectives of residents, their families, and nursing home employees during the COVID-19 pandemic. A scoping review was carried out using the PRISMA Extension for Scoping Reviews. The inclusion criteria were: qualitative and/or mixed methods studies in English, French, Portuguese, and Spanish. The review covers studies published from 11 March 2020 to 15 February 2021. CINAHL, PubMed, Web of Science, ScienceDirect, Scopus, British Nursing Index, Proquest, PsycInfo, and Google Scholar databases were used. We conducted a systematic narrative synthesis, presenting the results narratively and showing descriptive statistics on the studies reviewed. Sixteen documents were obtained from 175 results. Two studies focused on residents and one on their families. The remaining studies looked at professionals. Nursing homes had great difficulty managing resources, which was exacerbated by emotional exhaustion among residents, employees, and family members. In nursing homes, creative initiatives and new forms of leadership appeared to meet emerging needs during the pandemic. The results of the study show the impact of the pandemic on nursing homes and the response capacity present among residents, family members, and professionals
Study of the influence of process parameters on liquid andsupercritical CO2 extraction of oil from rendered materials: fish meal and oil characterization
tLiquid and supercritical CO2has been used to extract the remaining fat content from rendered fish meal.The effect of pressure (10–40 MPa) and temperature (25–80◦C) on the extraction kinetics and extractionyield has been investigated as well as the effect on the rendered fish meal. The extraction curves areinitially linear with a slope close to the oil solubility value in pressurized CO2. Based on previous fishoil solubility data reported in the literature, a general equation has been proposed to correlate fish oilsolubility data as a function of temperature and density of CO2. Fish meal has been characterized beforeand after extraction by determining the fat and protein content and its colour. Toxic trace elements havebeen also determined by ICP-MS in the fish meal showing that most of the toxic elements remained inthe fish meal after extraction. Characterization of extracted oil was also performed by determining thefatty acid group composition and some physical parameters such as colourSpanish Government through MINECO (CTQ2012-39131-C02-01). To SARVAL for supplying the fish meal. S.L.B.acknowledges support by the Secretary of Education Public ofMexico for the fellowship through PROMEP program and Tech-nological University of Moreli
The overlooked immune state in candidemia: A risk factor for mortality
Producción CientíficaLymphopenia has been related to increased mortality in septic patients. Nonetheless, the impact of lymphocyte count on candidemia mortality and prognosis has not been addressed. We conducted a retrospective study, including all admitted patients with candidemia from 2007 to 2016. We examined lymphocyte counts during the first 5 days following the diagnosis of candidemia. Multivariable logistic regression analysis was performed to determine the relationship between lymphocyte count and mortality. Classification and Regression Tree analysis was used to identify the best cut-off of lymphocyte count for mortality associated with candidemia. From 296 cases of candidemia, 115 died, (39.8% 30-day mortality). Low lymphocyte count was related to mortality and poor outcome (p < 0.001). Lymphocyte counts <0.703 × 109 cells/L at diagnosis (area under the curve (AUC)-ROC, 0.783 ± 0.042; 95% confidence interval (CI), 0.700–0.867, p < 0.001), and lymphocyte count <1.272 × 109 cells/L five days later (AUC-ROC, 0.791 ± 0.038; 95%CI, 0.716–0.866, p < 0.001) increased the odds of mortality five-fold (odds ratio (OR), 5.01; 95%CI, 2.39–10.93) at time of diagnosis, and three-fold (OR, 3.27; 95%CI, 1.24–8.62) by day 5, respectively. Low lymphocyte count is an independent predictor of mortality in patients with candidemia and might serve as a biomarker for predicting candidemia-associated mortality and poor outcome.Junta de Castilla y León (grant VA161G18
Evaluation of the potential association of SOHLH2 polymorphisms with non-obstructive azoospermia susceptibility in a large European population
Non-obstructive azoospermia (NOA) or spermatogenic failure is a complex disease with an important genetic component that causes infertility in men. Known genetic factors associated with NOA include AZF microdeletions of the Y chromosome or karyotype abnormalities; however, most causes of NOA are idiopathic. During the last decade, a large list of associations between single-nucleotide polymorphisms (SNP) and NOA have been reported. However, most of the genetic studies have been performed only in Asian populations. We aimed to evaluate whether the previously described association in Han Chinese between NOA and two SNPs of the SOHLH2 gene (involved in the spermatogenesis process) may also confer risk for NOA in a population of European ancestry. We genotyped a total of 551 NOA patients (218 from Portugal and 333 from Spain) and 1,050 fertile controls (226 from Portugal and 824 from Spain) for the genetic variants rs1328626 and rs6563386 using TaqMan assays. To test for association, we compared the allele and genotype frequencies between cases and controls using an additive model. A haplotype analysis and a meta-analysis using the inverse variance method with our data and those of the original Asian study were also performed. No statistically significant differences were observed in any of the analyses described above. Therefore, considering the high statistical power of our study, it is not likely that the two analysed SOHLH2 genetic variants are related with an increase susceptibility to NOA in the European population.info:eu-repo/semantics/publishedVersio
High p16 expression and heterozygous RB1 loss are biomarkers for CDK4/6 inhibitor resistance in ER+ breast cancer
Breast cancer; Cancer models; Predictive markersCáncer de mama; Modelos de cáncer; Marcadores predictivosCàncer de pulmó; Models de càncer; Marcadors predictiusCDK4/6 inhibitors combined with endocrine therapy have demonstrated higher antitumor activity than endocrine therapy alone for the treatment of advanced estrogen receptor-positive breast cancer. Some of these tumors are de novo resistant to CDK4/6 inhibitors and others develop acquired resistance. Here, we show that p16 overexpression is associated with reduced antitumor activity of CDK4/6 inhibitors in patient-derived xenografts (n = 37) and estrogen receptor-positive breast cancer cell lines, as well as reduced response of early and advanced breast cancer patients to CDK4/6 inhibitors (n = 89). We also identified heterozygous RB1 loss as biomarker of acquired resistance and poor clinical outcome. Combination of the CDK4/6 inhibitor ribociclib with the PI3K inhibitor alpelisib showed antitumor activity in estrogen receptor-positive non-basal-like breast cancer patient-derived xenografts, independently of PIK3CA, ESR1 or RB1 mutation, also in drug de-escalation experiments or omitting endocrine therapy. Our results offer insights into predicting primary/acquired resistance to CDK4/6 inhibitors and post-progression therapeutic strategies
Un viaje al Cosmos en 52 semanas
230 p.Este libro se plantea como una serie de artículos que dibujan un recorrido
por el Universo, desde lo más cercano a lo más distante. Constituye una herramienta útil y actualizada para los interesados en la astronomía, y combina el conocimiento básico con los resultados científicos más novedosos. La astrofísica constituye una ciencia viva y en permanente avance por ello nos encontramos con un Plutón que ya no es considerado planeta; con nuevos datos sobre la posible presencia de agua en Marte; con géiseres en Encelado, un pequeño satélite de Saturno que se creía
geológicamente inactivo; con una miríada de nuevos planetas girando alrededor
de otras estrellas; con, quizá, un nuevo tipo de agujero negro y fascinantes
resultados sobre las explosiones cortas de rayos gamma, uno de los eventos más
energéticos del Universo y, hasta hace poco, también uno de los más
desconocidos; con la mision COROT, y otras, como BepiColombo, que ya se encuentran en su fase de desarrollo instrumental.Peer reviewe
Common genetic variation in KATNAL1 non-coding regions is involved in the susceptibility to severe phenotypes of male infertility
Free PMC article: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9546047/Background: Previous studies in animal models evidenced that genetic mutations
of KATNAL1, resulting in dysfunction of its encoded protein, lead to male infertility
through disruption of microtubule remodelling and premature germ cell exfoliation.
Subsequent studies in humans also suggested a possible role of KATNAL1 single nucleotide polymorphisms in the development of male infertility as a consequence of
severe spermatogenic failure.
Objectives: The main objective of the present study is to evaluate the effect of the
common genetic variation of KATNAL1 in a large and phenotypically well-characterised
cohort of infertile men because of severe spermatogenic failure.
Materials and methods: A total of 715 infertile men because of severe spermato genic failure, including 210 severe oligospermia and 505 non-obstructive azoospermia
patients, as well as 1058 unaffected controls were genotyped for three KATNAL1
single-nucleotide polymorphism taggers (rs2077011, rs7338931 and rs2149971).
Case–control association analyses by logistic regression assuming different models
and in silico functional characterisation of risk variants were conducted.
Results: Genetic associations were observed between the three analysed taggers and
different severe spermatogenic failure groups. However, in all cases, the haplotype
model (rs2077011*C | rs7338931*T | rs2149971*A) better explained the observed
associations than the three risk alleles independently. This haplotype was associated
with non-obstructive azoospermia (adjusted p = 4.96E-02, odds ratio = 2.97), Sertoli cell only syndrome (adjusted p = 2.83E-02, odds ratio = 5.16) and testicular sperm
extraction unsuccessful outcomes (adjusted p = 8.99E-04, odds ratio = 6.13). The in
silico analyses indicated that the effect on severe spermatogenic failure predisposition
could be because of an alteration of the KATNAL1 splicing pattern.
Conclusions: Specific allelic combinations of KATNAL1 genetic polymorphisms may
confer a risk of developing severe male infertility phenotypes by favouring the
overrepresentation of a short non-functional transcript isoform in the testis.This work was supported by the Spanish Ministry of Economy and
Competitiveness through the Spanish National Plan for Scientific
and Technical Research and Innovation (refs. SAF2016-78722-R and
PID2020-120157RB-I00), the ‘Instituto de Salud Carlos III’ (Fondo
de Investigaciones Sanitarias)/Fondo Europeo de Desarrollo Regional
‘Una manera de hacer Europa’ (FIS/FEDER) (ref. DTS18/00101 to
Sara Larriba), the Generalitat de Catalunya (ref. 2017SGR191), the
‘Ramón y Cajal’ program (ref. RYC-2014-16458) and the ‘Juan de
la Cierva Incorporación’ program (ref. IJC2018-038026-I), as well as
the Andalusian Government through the R&D&i Projects Grants for
Universities and Public Research Entities (ref. PY20_00212), which
include FEDER funds. Andrea Guzmán-Jiménez was a recipient of a
grant from the Spanish Ministry of Education and Professional Training (‘Becas de Colaboración en Departamentos Universitarios para el curso académico 2020/2021’). Patricia I. Marques is supported by
the FCT post-doctoral fellowship (SFRH/BPD/120777/2016), financed
from the Portuguese State Budget of the Ministry for Science, Tech nology and High Education and from the European Social Fund,
available through the ‘Programa Operacional do Capital Humano’. João
Gonçalves was partially funded by FCT/MCTES through national funds
attributed to the Centre for Toxicogenomics and Human Health—
ToxOmics (UID/BIM/00009/2016 and UIDB/00009/2020). Sara Larriba is sponsored by the Researchers Consolidation Program (ISCIII
SNS/Dpt. Salut Generalitat de Catalunya) (CES09/020).info:eu-repo/semantics/publishedVersio
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