Seminal plasma modulates miRNA expression by sow genital tract lining explants

Research Areas: Biochemistry & Molecular BiologyThe seminal plasma (SP) modulates the female reproductive immune environment after mating, and microRNAs (miRNAs) could participate in the process. Considering that the boar ejaculate is built by fractions di ering in SP-composition, this study evaluated whether exposure of mucosal explants of the sow internal genital tract (uterus, utero-tubal junction and isthmus) to di erent SP-fractions changed the profile of explant-secreted miRNAs. Mucosal explants retrieved from oestrus sows (n = 3) were in vitro exposed to: Medium 199 (M199, Control) or M199 supplemented (1:40 v/v) with SP from the sperm-rich fraction (SRF), the post-SRF or the entire recomposed ejaculate, for 16 h. After, the explants were cultured in M199 for 24 h to finally collect the media for miRNA analyses using GeneChip miRNA 4.0 Array (A ymetrix). Fifteen di erentially expressed (False Discovery Rate (FDR) < 0.05 and Fold-change 2) miRNAs (11 down- versus 4 up-regulated) were identified (the most in the media of uterine explants incubated with SP from post-SRF). Bioinformatics analysis identified that predicted target genes of dysregulated miRNAs, mainly miR-34b, miR-205, miR-4776-3p and miR-574-5p, were involved in functions and pathways related to immune response. In conclusion, SP is able to elicit changes in the miRNAs profile secreted by female genital tract, ultimately depending SP-composition.info:eu-repo/semantics/publishedVersio

Real-life disease monitoring in follicular lymphoma patients using liquid biopsy ultra-deep sequencing and PET/CT

In the present study, we screened 84 Follicular Lymphoma patients for somatic mutations suitable as liquid biopsy MRD biomarkers using a targeted next-generation sequencing (NGS) panel. We found trackable mutations in 95% of the lymph node samples and 80% of the liquid biopsy baseline samples. Then, we used an ultra-deep sequencing approach with 2 · 10−4 sensitivity (LiqBio-MRD) to track those mutations on 151 follow-up liquid biopsy samples from 54 treated patients. Positive LiqBio-MRD at first-line therapy correlated with a higher risk of progression both at the interim evaluation (HRINT 11.0, 95% CI 2.10–57.7, p = 0.005) and at the end of treatment (HREOT, HR 19.1, 95% CI 4.10–89.4, p < 0.001). Similar results were observed by PET/CT Deauville score, with a median PFS of 19 months vs. NR (p < 0.001) at the interim and 13 months vs. NR (p < 0.001) at EOT. LiqBio-MRD and PET/CT combined identified the patients that progressed in less than two years with 88% sensitivity and 100% specificity. Our results demonstrate that LiqBio-MRD is a robust and non-invasive approach, complementary to metabolic imaging, for identifying FL patients at high risk of failure during the treatment and should be considered in future response-adapted clinical trials.This study has been funded by Instituto de Salud Carlos III (ISCIII) and co-funded by the European Union through the projects PI21/00314, PI19/01430, PI19/01518 and PI18/00295, PTQ2020-011372, CP19/00140, CP22/00082, Doctorado industrial CAM IND2020/TIC-17402 and CRIS cancer foundation

Social mobility and healthy behaviours from a gender perspective in the Spanish multicase-control study (MCC-Spain)

There is evidence for the influence of socioeconomic status (SES) on healthy behaviours but the effect of social mobility (SM) is not yet well known. This study aims to analyse the influence of origin and destination SES (O-SES and D-SES) and SM on healthy behaviours and co-occurrence, from an integrated gender and age perspective. Data were obtained from the controls of MCC-Spain between 2008-2013 (3,606 participants). Healthy behaviours considered: healthy diet, moderate alcohol consumption, non-smoking and physical activity. SM was categorized as stable high, upward, stable medium, downward or stable low. Binary and multinomial logistic regression models were adjusted. Those aged <65, with a low O-SES, D-SES and stable low SM are less likely to have healthy behaviours in the case of both women (physically active: OR = 0.65 CI = 0.45-0.94, OR = 0.71 CI = 0.52-0.98, OR = 0.61 CI = 0.41-0.91) and men (non-smokers: OR = 0.44 CI = 0.26-0.76, OR = 0.54 CI = 0.35-0.83, OR = 0.41 CI 0.24-0.72; physically active: OR = 0.57 CI = 0.35-0.92, OR = 0.64 CI = 0.44-0.95, OR = 0.53 CI = 0.23-0.87). However, for those aged ≥65, this probability is higher in women with a low O-SES and D-SES (non-smoker: OR = 8.09 CI = 4.18-15.67, OR = 4.14 CI = 2.28-7.52; moderate alcohol consumption: OR = 3.00 CI = 1.45-6.24, OR = 2.83 CI = 1.49-5.37) and in men with a stable low SM (physically active: OR = 1.52 CI = 1.02-1.26). In the case of men, the same behaviour pattern is observed in those with a low O-SES as those with upward mobility, with a higher probability of co-occurring behaviours (three-to-four behaviours: OR = 2.00 CI = 1.22-3.29; OR = 3.13 CI = 1.31-7.48). The relationship of O-SES, D-SES and SM with healthy behaviours is complex and differs according to age and gender.This research was supported by the “Acción Transversal del Cancer”, approved by the Spanish Council of Ministers on 11th October 2007, by the Instituto de Salud Carlos III-FEDER [grant number:PI08/1770, PI08/0533, PI08/1359, PS09/00773-Cantabria, PS09/01286-León, PS09/01903-Valencia, PS09/02078-Huelva, PS09/ 01662-Granada, PI11/01403, PI11/01889-FEDER, PI11/00226, PI11/01810, PI11/02213, PI12/00488, PI12/00265, PI12/01270, PI12/00715, PI12/00150, PI14/01219, PI14/0613, PI15/00069, PI15/00914, PI15/01032, PI11/01810, PI14/01219, PI11/02213, PIE16/00049, PI17/01179, PI17-00092], by the Fundación Marqués de Valdecilla [grant number: API 10/09], by the ICGC International Cancer Genome Consortium CLL (The ICGC CLL-Genome Project is funded by Spanish Ministerio de Economía y Competitividad (MINECO) through the Instituto de Salud Carlos III (ISCIII)), by the Red Temática de Investigación del Cáncer (RTICC) del ISCIII [grant number: RD12/0036/0036], by the Junta de Castilla y León [grant number: LE22A10-2], by the Consejería de Salud of the Junta de Andalucía [grant number: PI-0571-2009, PI-0306-2011, salud201200057018tra], by the Conselleria de Sanitat of the Generalitat Valenciana [grant number: AP_061/10], by the Recercaixa [grant number: 2010ACUP00310], by the Regional Government of the Basque Country, by the Consejería de Sanidad de la Región de Murcia, by the European Commission [grant number: FOOD-CT-2006-036224-HIWATE], by the Spanish Association Against Cancer (AECC) Scientific Foundation [grant number: GCTRA18022MORE], by the Catalan Government-Agency for Management of University and Research Grants (AGAUR) [grant number: 2014SGR647, 2014SGR850 and 2017SGR723], by the Fundación Caja de Ahorros de Asturias and by the University of Oviedo. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.S

Detrimental Effects of Non-Functional Spermatozoa on the Freezability of Functional Spermatozoa from Boar Ejaculate

In the present study, the impact of non-functional spermatozoa on the cryopreservation success of functional boar spermatozoa was evaluated. Fifteen sperm-rich ejaculate fractions collected from five fertile boars were frozen with different proportions of induced non-functional sperm (0 –native semen sample-, 25, 50 and 75% non-functional spermatozoa). After thawing, the recovery of motile and viable spermatozoa was assessed, and the functional of the spermatozoa was evaluated from plasma membrane fluidity and intracellular reactive oxygen species (ROS) generation upon exposure to capacitation conditions. In addition, the lipid peroxidation of the plasma membrane was assessed by the indirect measurement of malondialdehyde (MDA) generation. The normalized (with respect to a native semen sample) sperm motility (assessed by CASA) and viability (cytometrically assessed after staining with Hoechst 33342, propidium iodide and fluorescein-conjugated peanut agglutinin) decreased (p<0.01) as the proportion of functional spermatozoa in the semen samples before freezing decreased, irrespective of the semen donor. However, the magnitude of the effect differed (p<0.01) among boars. Moreover, semen samples with the largest non-functional sperm subpopulation before freezing showed the highest (p<0.01) levels of MDA after thawing. The thawed viable spermatozoa of semen samples with a high proportion of non-functional spermatozoa before freezing were also functionally different from those of samples with a low proportion of non-functional spermatozoa. These differences consisted of higher (p<0.01) levels of intracellular ROS generation (assessed with 5-(and-6) chloromethyl-20,70-dichlorodihydrofluorescein diacetate acetyl ester; CM-H2DCFDA) and increased (p<0.01) membrane fluidity (assessed with Merocyanine 540). These findings indicate that non-functional spermatozoa in the semen samples before freezing negatively influence the freezability of functional spermatozoa

Early Developing Pig Embryos Mediate Their Own Environment in the Maternal Tract

The maternal tract plays a critical role in the success of early embryonic development providing an optimal environment for establishment and maintenance of pregnancy. Preparation of this environment requires an intimate dialogue between the embryo and her mother. However, many intriguing aspects remain unknown in this unique communication system. To advance our understanding of the process by which a blastocyst is accepted by the endometrium and better address the clinical challenges of infertility and pregnancy failure, it is imperative to decipher this complex molecular dialogue. The objective of the present work is to define the local response of the maternal tract towards the embryo during the earliest stages of pregnancy. We used a novel in vivo experimental model that eliminated genetic variability and individual differences, followed by Affymetrix microarray to identify the signals involved in this embryo-maternal dialogue. Using laparoscopic insemination one oviduct of a sow was inseminated with spermatozoa and the contralateral oviduct was injected with diluent. This model allowed us to obtain samples from the oviduct and the tip of the uterine horn containing either embryos or oocytes from the same sow. Microarray analysis showed that most of the transcripts differentially expressed were down-regulated in the uterine horn in response to blastocysts when compared to oocytes. Many of the transcripts altered in response to the embryo in the uterine horn were related to the immune system. We used an in silico mathematical model to demonstrate the role of the embryo as a modulator of the immune system. This model revealed that relatively modest changes induced by the presence of the embryo could modulate the maternal immune response. These findings suggested that the presence of the embryo might regulate the immune system in the maternal tract to allow the refractory uterus to tolerate the embryo and support its development

The evolution of the ventilatory ratio is a prognostic factor in mechanically ventilated COVID-19 ARDS patients

Background: Mortality due to COVID-19 is high, especially in patients requiring mechanical ventilation. The purpose of the study is to investigate associations between mortality and variables measured during the first three days of mechanical ventilation in patients with COVID-19 intubated at ICU admission. Methods: Multicenter, observational, cohort study includes consecutive patients with COVID-19 admitted to 44 Spanish ICUs between February 25 and July 31, 2020, who required intubation at ICU admission and mechanical ventilation for more than three days. We collected demographic and clinical data prior to admission; information about clinical evolution at days 1 and 3 of mechanical ventilation; and outcomes. Results: Of the 2,095 patients with COVID-19 admitted to the ICU, 1,118 (53.3%) were intubated at day 1 and remained under mechanical ventilation at day three. From days 1 to 3, PaO2/FiO2 increased from 115.6 [80.0-171.2] to 180.0 [135.4-227.9] mmHg and the ventilatory ratio from 1.73 [1.33-2.25] to 1.96 [1.61-2.40]. In-hospital mortality was 38.7%. A higher increase between ICU admission and day 3 in the ventilatory ratio (OR 1.04 [CI 1.01-1.07], p = 0.030) and creatinine levels (OR 1.05 [CI 1.01-1.09], p = 0.005) and a lower increase in platelet counts (OR 0.96 [CI 0.93-1.00], p = 0.037) were independently associated with a higher risk of death. No association between mortality and the PaO2/FiO2 variation was observed (OR 0.99 [CI 0.95 to 1.02], p = 0.47). Conclusions: Higher ventilatory ratio and its increase at day 3 is associated with mortality in patients with COVID-19 receiving mechanical ventilation at ICU admission. No association was found in the PaO2/FiO2 variation

Avances de la técnica de preselección del sexo en el ganado porcino mediante separación de espermatozoides X e Y por citometría de flujo

La separación espermática por citometría de flujo de los espermatozoides X e Y, es actualmente el único método eficaz para la obtención de descendencia de sexo deseado. En porcino, la selección del sexo de las camadas incrementaría notablemente los beneficios productivos de las explotaciones productoras de animales de alto valor genético al poder manejar los esquemas de selección, desviándolos hacia un sexo u otro según las necesidades de cada explotación. Recientemente, se han realizado avances importantes en la separación espermática mediante esta técnica en porcino. Sin embargo, es necesario un mayor conocimiento de diferentes aspectos relacionados con la eficiencia del proceso, con el efecto del procedimiento sobre la viabilidad y la capacidad fecundante de estos espermatozoides, así como del posible efecto perjudicial del propio proceso de separación sobre el ADN de los espermatozoides de verraco. Esta tesis recoge resultados derivados de experiencias destinadas a estudiar estos aspectos

Estructura y evolución del comercio exterior de Mercosur

En el presente trabajo se analiza la estructura y evolución del comercio exterior del Mercado Común del Sur (Mercosur) con sus principales socios comerciales en el período 1961-1998. El objetivo principal es evaluar la competitividad internacional de Mercosur desde un enfoque sectorial, basado en el análisis de la ventaja comparativa dinámica para los distintos sectores productivos y en el cálculo de distintos índices de comercio intraindustrial para medir la importancia de este tipo de comercio en los intercambios comerciales totales con la Unión Europea, Estados Unidos, Canadá y Japón. Se comprueba que existe una marcada especialización de Mercosur en productos básicos y una fuerte dependencia del exterior en productos con alto contenido tecnológico. Por último, los niveles de comercio intraindustrial son mucho más bajos que los existentes en la Unión Europea o, en general, entre países desarrollados

INMUNOLOCALIZACIÓN DE LA ESPERMADHESINA PSP-I/PSP-II SOBRE LOS ESPERMATOZOIDES DE VERRACO

The heterodimeric PSP-I/PSP-II spermadhesin represents the major protein of boar seminal plasma. We investigated the pattern of binding of PSP-I/PSP-II on boar spermatozoa, from the epididymis to the site of fertilization. Spermatozoa were collected from the cauda epididymis, from ejaculates of mature boars,and from the oviduct of sows mated with a fertile boar and slaughtered 6-8 h after spontaneous ovulation. Immunolocalization of the PSP-I/PSP-II spermadhesin on spermatozoa was studied using a monospecific polyclonal antibody raised against the PSP-II subunit in rabbits. A 10-nm gold-labelled goat anti-rabbit IgG was used as the secondary antibody and the samples subjected to silver enhancement. Light and scanning electron microscopy were used to examine the distribution of PSP-I/II on spermatozoa. Epididymal spermatozoa were consistently immunonegative. In contrast, ejaculated spermatozoa exhibited large aggregates of silver coating the acrosomal cap. When spermatozoa were collected from the oviduct, two sperm populations, either with or without PSP-I/II on the acrosomal cap, were found tempting to considered capacitation as involved in the release/removal of PSP-I/II from the sperm surface. The results suggest PSP-II/PSP-II binds to the sperm surface at ejaculation, and that its release might be related to pre-fertilization changes of the sperm membrane.El heterodímero PSP-I/PSP-II es el mayor componente proteico del plasma seminal del verraco. En este trabajo se investigó el patrón de unión del heterodímero PSP-I/PSP-II sobre los espermatozoides de verraco desde su almacenamiento en el epidídimo hasta el lugar de la fecundación. Para ello se recogieron espermatozoides procedentes de la cola del epidídimo, del eyaculado de verracos maduros sexualmente y del oviducto de cerdas multíparas las cuales habían sido cubiertas por un verraco y sacrificadas aproximadamente 6-8 horas tras la ovulación. La inmunolocalización se realizó con anticuerpos policlonales monoespecífico anti-PSP-II obtenidos mediante inmunización de conejas por inyecciones subcutaneas. Se uso como anticuerpo secundario un anticuerpo de cabra anti-conejo marcado con partículas de oro de 10 nm. Tras la incubación con el anticuerpo secundario las muestras fueron sometidas a amplificación con plata. Para el examen de la distribución del heterodímero PSP-I/PSP-II se usó tanto microscopía óptica como microscopía electrónica de transmisión. Los espermatozoides procedentes del epidídimo fueron inmunonegativos de manera constante. En cambio, cuando se examinaron los espermatozoides procedentes del eyaculado, estos mostraron grandes agregados de plata recubriendo la región acrosomal del espermatozoide. Cuando los espermatozoides provenían del oviducto nos encontramos dos subpoblaciones espermáticas, con o sin el heterodímero PSP-I/PSP-II sobre la región acrosomal, haciéndonos pensar que la capacitación espermática podría estar relacionada con la eliminación del heterodímero PSP-I/PSP-II de la superficie del espermatozoide. En conclusión, nuestros resultados sugieren que el heterodímero PSP-I/PSP-II se une al espermatozoide y su liberación podría estar relacionada con cambios en la membrana plasmática del espermatozoide previos a la fecundación