Generation of a novel three-dimensional scaffold-based model of the bovine endometrium

Bovine in vitro endometrial models that resemble tissue function in vivo are needed to study infertility, long-term uterine alterations induced by pathogens and impact of endocrine disruptor chemicals on reproductive function and other reproductive system complications that cause high economic losses in livestock species. The present study aimed to generate an innovative, reproducible, and functional 3D scaffold-based model of the bovine endometrium structurally robust for long term-culture. We developed a multicellular model containing both endometrial epithelial and stromal cells. Epithelial cells organized to form a luminal-like epithelial layer on the surface of the scaffold. Stromal cells produced their own extracellular matrix forming a stable subepithelial compartment that physiologically resembles the normal endometrium. Both cell types released prostaglandin E2 and prostaglandin F2α following a treatment with oxytocin and arachidonic acid. Additionally signal pathways mediating oxytocin and arachidonic acid stimulation of prostaglandin synthesis were analyzed by real time PCR (RT-PCR). Oxytocin receptor (OXTR), prostaglandin E2 receptor 2 (EP2), prostaglandin E2 receptor 4 (EP4), prostaglandin F receptor (PTGFR), prostaglandin E synthase (PTGES), PGF-synthase (PGFS) and prostaglandin-endoperoxide synthase 2 (COX-2) expression was detected in both control and treatment groups, however, only significant changes in abundance of OXTR mRNA transcripts were found. The results obtained by this study are a step forward in bovine in vitro culture technology. This 3D scaffold-based model provides a platform to study regulatory mechanisms involved in endometrial physiology and can set the basis for a broader tool for designing and testing novel therapeutic strategies for recurrent uterine pathologies

SARS-CoV-2 Breakthrough Infections: Incidence and Risk Factors in a Large European Multicentric Cohort of Health Workers.

Background: The research aimed to investigate the incidence of SARS-CoV-2 breakthrough infections and their determinants in a large European cohort of more than 60,000 health workers. Methods: A multicentric retrospective cohort study, involving 12 European centers, was carried out within the ORCHESTRA project, collecting data up to 18 November 2021 on fully vaccinated health workers. The cumulative incidence of SARS-CoV-2 breakthrough infections was investigated with its association with occupational and social-demographic characteristics (age, sex, job title, previous SARS-CoV-2 infection, antibody titer levels, and time from the vaccination course completion). Results: Among 64,172 health workers from 12 European health centers, 797 breakthrough infections were observed (cumulative incidence of 1.2%). The primary analysis using individual data on 8 out of 12 centers showed that age and previous infection significantly modified breakthrough infection rates. In the meta-analysis of aggregated data from all centers, previous SARS-CoV-2 infection and the standardized antibody titer were inversely related to the risk of breakthrough infection (p = 0.008 and p = 0.007, respectively). Conclusion: The inverse correlation of antibody titer with the risk of breakthrough infection supports the evidence that vaccination plays a primary role in infection prevention, especially in health workers. Cellular immunity, previous clinical conditions, and vaccination timing should be further investigated

Efficiency of enzymes and benzalkonium chloride treatments against Listeria monocytogenes dual-species biofilms determined by fluorescence microscopy and image analysis

In the present study, three different mixed-species Listeria monocytogenes-carrying biofilms previously isolated from surfaces of seafood, meat and dairies industrial environments were selected in order to test the efficiency of cleaning and disinfection procedures based on the combination of an enzymatic solution, followed by a treatment with benzalkonium chloride (BAC). Biofilm samples were cultured onto AISI-316 type stainless steel 1 cm2 coupons in a batch system at 25 degrees and the effect of sequential exposure to different concentrations of Pronase – BAC, Cellulase – BAC or DNAseI – BAC solutions was then assessed in mature biofilms after 168 hours in nine different experiments using an orthogonal first order approach. Treated samples were stained using then LIVE/DEAD Biofilm viability kit (Invitrogen) and the efficiency of each individual treatment was then determined by epifluorescence microscopy and subsequent image analysis of 25-field mosaics representing a total surface of 1.92mm2, so as to quantify the remaining living cells attached to the surface of the coupon. Analysis of data reveals that synergistic effects are observed after using Pronase – BAC on biofilms coming from fish and dairy industries obtaining almost a total cell detachment in the latter case, whereas DNAse – BAC treatments are efficient only against fish industry biofilms. No significant effect was observed among the Cellulase – BAC procedures, being in some cases even counter-productive. In addition to this, individual effects of Cellulase and Pronase are observed on fish and fish and dairy industry biofilms respectively demonstrating its potential as a cleaning agent even at low concentrations of the enzyme. These results show how the usage of customised treatments combining commercially available enzymes with classical chemically-based disinfection procedures may represent an attractive and efficient alternative against bacterial biofilms in food industry

Performance of the Finnish Diabetes Risk Score and a Simplified Finnish Diabetes Risk Score in a Community-Based, Cross-Sectional Programme for Screening of Undiagnosed Type 2 Diabetes Mellitus and Dysglycaemia in Madrid, Spain: The SPREDIA-2 Study

<div><p>Aim</p><p>To evaluate the performance of the Finnish Diabetes Risk Score (FINDRISC) and a simplified FINDRISC score (MADRISC) in screening for undiagnosed type 2 diabetes mellitus (UT2DM) and dysglycaemia.</p><p>Methods</p><p>A population-based, cross-sectional, descriptive study was carried out with participants with UT2DM, ranged between 45–74 years and lived in two districts in the north of metropolitan Madrid (Spain). The FINDRISC and MADRISC scores were evaluated using the area under the receiver operating characteristic curve method (ROC-AUC). Four different gold standards were used for UT2DM and any dysglycaemia, as follows: fasting plasma glucose (FPG), oral glucose tolerance test (OGTT), HbA1c, and OGTT or HbA1c. Dysglycaemia and UT2DM were defined according to American Diabetes Association criteria.</p><p>Results</p><p>The study population comprised 1,426 participants (832 females and 594 males) with a mean age of 62 years (SD = 6.1). When HbA1c or OGTT criteria were used, the prevalence of UT2DM was 7.4% (10.4% in men and 5.2% in women; p<0.01) and the FINDRISC ROC-AUC for UT2DM was 0.72 (95% CI, 0.69–0.74). The optimal cut-off point was ≥13 (sensitivity = 63.8%, specificity = 65.1%). The ROC-AUC of MADRISC was 0.76 (95% CI, 0.72–0.81) with ≥13 as the optimal cut-off point (sensitivity = 84.8%, specificity = 54.6%). FINDRISC score ≥12 for detecting any dysglycaemia offered the best cut-off point when HbA1c alone or OGTT and HbA1c were the criteria used.</p><p>Conclusions</p><p>FINDRISC proved to be a useful instrument in screening for dysglycaemia and UT2DM. In the screening of UT2DM, the simplified MADRISC performed as well as FINDRISC.</p></div

Characteristics of the study participants.

<p>Characteristics of the study participants.</p

Characteristics of the FINDRISC questionnaire using different cut-off values for dysglycaemia.

<p>Characteristics of the FINDRISC questionnaire using different cut-off values for dysglycaemia.</p

Receiver operating characteristic curves in patients with newly diagnosed diabetes based on three diagnostic criteria: HbA1c (A), OGTT (B), HbA1c and OGTT (C), and FPG (D).

<p>Receiver operating characteristic curves in patients with newly diagnosed diabetes based on three diagnostic criteria: HbA1c (A), OGTT (B), HbA1c and OGTT (C), and FPG (D).</p

Characteristics of the simplified FINDRISC (MADRISC) questionnaire using different cut-off values for undiagnosed diabetes mellitus.

<p>Characteristics of the simplified FINDRISC (MADRISC) questionnaire using different cut-off values for undiagnosed diabetes mellitus.</p