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Genomic insights into drug resistance and virulence platforms, CRISPR-Cas systems and phylogeny of commensal E. Coli from wildlife

Commensal bacteria act as important reservoirs of virulence and resistance genes. However, existing data are generally only focused on the analysis of human or human-related bacterial populations. There is a lack of genomic studies regarding commensal bacteria from hosts less exposed to antibiotics and other selective forces due to human activities, such as wildlife. In the present study, the genomes of thirty-eight E. coli strains from the gut of various wild animals were sequenced. The analysis of their accessory genome yielded a better understanding of the role of the mobilome on inter-bacterial dissemination of mosaic virulence and resistance plasmids. The study of the presence and composition of the CRISPR/Cas systems in E. coli from wild animals showed some viral and plasmid sequences among the spacers, as well as the relationship between CRISPR/Cas and E. coli phylogeny. Further, we constructed a single nucleotide polymorphisms-based core tree with E. coli strains from different sources (humans, livestock, food and extraintestinal environments). Bacteria from humans or highly human-influenced settings exhibit similar genetic patterns in CRISPR-Cas systems, plasmids or virulence/resistance genes-carrying modules. These observations, together with the absence of significant genetic changes in their core genome, suggest an ongoing flow of both mobile elements and E. coli lineages between human and natural ecosystems.Funding: This work was partially supported by project SAF2016-76571-R and PID2019-106158RB-I00 from the Agencia Estatal de Investigación (AEI) of Spain and the Fondo Europeo de Desarrollo Regional (FEDER) of EU. During the experimental work of this study, Carla Andrea Alonso Alonso had a predoctoral fellowship FPI from MINECO

Evaluation of sperm integrin α5β1 as a potential marker of fertility in humans

Sperm selection for assisted reproduction techniques is generally based on basic parameters, while key aspects of sperm competence and its journey from the deposition site to the fertilization site are overlooked. Consequently, identifying molecular markers in spermatozoa that can efficiently predict the fertility of a semen sample could be of great interest, particularly in cases of idiopathic male infertility. When spermatozoa reach the female reproductive tract, it provides to them the cellular and molecular microenvironment needed to acquire fertilizing ability. In this sense, considering the role that integrin α5β1 of spermatozoa plays in reproduction-related events, we investigated the correlation between the subcellular localization of sperm integrin α5β1 and early embryo development outcome after in vitro fertilization (IVF) procedures in human. Twenty-four semen samples from normozoospermic men and metaphase II (MII) oocytes from healthy women aged under 38 years, from couples who underwent IVF cycles, were used in this work. Sperm α5β1 localization was evaluated by immunofluorescence assay using an antibody against integrin α5 subunit. Integrin α5β1 was mainly localized in the sperm acrosomal region (45.33±7.89%) or the equatorial segment (30.12±7.43%). The early embryo development rate (data obtained from the Fertility Center) correlated positively with the localization of α5β1 in the acrosomal region (number of usable embryos / inseminated oocytes: ρ = 0.75; p<0.01 and number of usable embryos/total number of two pronuclear zygotes: ρ = 0.80; p<0.01). However, this correlation was not significant when the equatorial segment mark was evaluated. In addition, human sperm released from co-culture with bovine oviductal epithelial cells (BOEC) showed a significant enrichment in the acrosomal localization pattern of α5β1 compared to those sperm that were not co-cultured with BOEC (85.20±5.35% vs 35.00±17.09%, respectively, p<0.05). In conclusion, the evaluation of sperm integrin α5β1 immunolocalization could be a useful tool to select sperm with fertilizing ability from human semen samples before IVF procedures.Fil: Vernaz, Zoilo José. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos Aires; ArgentinaFil: Lottero Leconte, Raquel María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Alonso, Carlos Agustín Isidro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Rio, Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Morales, Maia Florencia. Fecunditas Instituto de Medicina Reproductiva; ArgentinaFil: Arroyo Salvo, Camila Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Valiente, Carla Carolina. Fecunditas Instituto de Medicina Reproductiva; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Lovaglio Diez, María. Fecunditas Instituto de Medicina Reproductiva; ArgentinaFil: Bogetti, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Arenas, Gabriela. Fecunditas Instituto de Medicina Reproductiva; ArgentinaFil: Rey Valzacchi, Gaston Javier. Fecunditas Instituto de Medicina Reproductiva; ArgentinaFil: Perez Martinez, Silvina Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; Argentin

Hide and seek between Andromeda's halo, disk, and giant stream

Photometry in B, V (down to V ~ 26 mag) is presented for two 23' x 23' fields of the Andromeda galaxy (M31) that were observed with the blue channel camera of the Large Binocular Telescope during the Science Demonstration Time. Each field covers an area of about 5.1kpc x 5.1kpc at the distance of M31 ((m-M)o ~ 24.4 mag), sampling, respectively, a northeast region close to the M31 giant stream (field S2), and an eastern portion of the halo in the direction of the galaxy minor axis (field H1). The stream field spans a region that includes Andromeda's disk and the giant stream, and this is reflected in the complexity of the color magnitude diagram of the field. One corner of the halo field also includes a portion of the giant stream. Even though these demonstration time data were obtained under non-optimal observing conditions the B photometry, acquired in time-series mode, allowed us to identify 274 variable stars (among which 96 are bona fide and 31 are candidate RR Lyrae stars, 71 are Cepheids, and 16 are binary systems) by applying the image subtraction technique to selected portions of the observed fields. Differential flux light curves were obtained for the vast majority of these variables. Our sample includes mainly pulsating stars which populate the instability strip from the Classical Cepheids down to the RR Lyrae stars, thus tracing the different stellar generations in these regions of M31 down to the horizontal branch of the oldest (t ~ 10 Gyr) component.Comment: 59 pages, 26 figures, 12 tables, ApJ in pres

Survival time among patients who were diagnosed with tuberculosis, the precocious deaths and associated factors in southern Brazil

Background: A diagnosis of tuberculosis (TB) does not mean that the disease will be treated successfully, since death may occur even among those who are known to the health services. Here, we aimed to analyze patient survival time from the diagnosis of TB to death, precocious deaths, and associated factors in southern Brazil. Methods: We conducted a longitudinal study with patients who were diagnosed with TB and who died due to the disease between 2008 and 2015 in southern Brazil. The starting point for measuring survival time was the patient’s diagnosis date. Techniques for survival analysis were employed, including the Kaplan-Meier test and Cox’s regression. A mixed-effect model was applied for identifying the associated factors to precocious deaths. Hazard ratio (HR) and odds ratio (OR) with 95% confidence intervals (95% CI) were estimated. We defined p value <0.05 as statistically significant for all statistics applied. Results: One hundred forty-six patients were included in the survival analysis, observing a median survival time of 23.5 days. We observed that alcoholism (HR=1.55, 95% CI=1.04-2.30) and being male (HR=6.49, 95% CI=1.03-2.68) were associated with death. The chance of precocious death within 60 days was 10.48 times greater than the chance of early death within 30 days. Conclusion: Most of the deaths occurred within 2 months after the diagnosis, during the intensive phase of the treatment. The use of alcohol and gender were associated with death, revealing inequality between men and women. This study advanced knowledge regarding the vulnerability associated with mortality. These findings must be addressed to fill a gap in the care cascades for active TB and ensure equity in health.publishersversionpublishe

Cold atoms in space: community workshop summary and proposed road-map

We summarise the discussions at a virtual Community Workshop on Cold Atoms in Space concerning the status of cold atom technologies, the prospective scientific and societal opportunities offered by their deployment in space, and the developments needed before cold atoms could be operated in space. The cold atom technologies discussed include atomic clocks, quantum gravimeters and accelerometers, and atom interferometers. Prospective applications include metrology, geodesy and measurement of terrestrial mass change due to, e.g., climate change, and fundamental science experiments such as tests of the equivalence principle, searches for dark matter, measurements of gravitational waves and tests of quantum mechanics. We review the current status of cold atom technologies and outline the requirements for their space qualification, including the development paths and the corresponding technical milestones, and identifying possible pathfinder missions to pave the way for missions to exploit the full potential of cold atoms in space. Finally, we present a first draft of a possible road-map for achieving these goals, that we propose for discussion by the interested cold atom, Earth Observation, fundamental physics and other prospective scientific user communities, together with the European Space Agency (ESA) and national space and research funding agencies

Prevalence and genetic characteristics of Staphylococcus aureus CC398 isolates from invasive infections in spanish hospitals, focusing on the livestock-independent CC398-MSSA clade

Background: Livestock-associated (LA)-CC398-MRSA is closely related to pigs, being unfrequently detected in human invasive infections. CC398-MSSA is emerging in human invasive infections in some countries, but genetic and epidemiological characteristics are still scarcely reported. Objectives: To determine the prevalence of Staphylococcus aureus (SA) CC398, both MRSA and MSSA, among blood cultures SA isolates recovered in Spanish hospitals located in regions with different pig-farming densities (PD) and characterize the recovered isolates. Methods: One thousand twenty-two SA isolates (761 MSSA, 261 MRSA) recovered from blood cultures during 6–12 months in 17 Spanish hospitals (2018–2019) were studied. CC398 lineage identification, detection of spa-types, and antibiotic resistance, virulence and human immune evasion cluster (IEC) genes were analyzed by PCR/sequencing. Results: Forty-four CC398-MSSA isolates (4.3% of SA; 5.8% of MSSA) and 10 CC398-MRSA isolates (1% of SA; 3.8% of MRSA) were detected. Eleven spa-types were found among the CC398-MSSA isolates with t571 and t1451 the most frequent spa-types detected (75%). Most of CC398-MSSA isolates were Immune-Evasion-Cluster (IEC)-positive (88.6%), tetracycline-susceptible (95.5%) and erythromycin/clindamycin–inducible-resistant/erm(T)-positive (75%). No statistical significance was detected when the CC398-MSSA/MSSA rate was correlated to PD (pigs/km2) (p = 0.108). On the contrary, CC398-MRSA isolates were all IEC-negative, predominately spa-t011 (70%), and the CC398-MRSA/MRSA rate was significantly associated to PD (p < 0.005). Conclusion: CC398-MSSA is an emerging clade in invasive infections in Spanish hospitals. CC398-MRSA (mostly t011) and CC398-MSSA (mostly t571 and t1451) show important differences, possibly suggesting divergent steps in host-adaptation evolutionary processes. While CC398-MRSA is livestock-associated (lacking IEC-system), CC398-MSSA seems to be mostly livestock-independent, carrying human-adaptation markers.