Aims. We demonstrate the high multiplex advantage of crowded field 3D spectroscopy with the new integral field spectrograph MUSE by means of a spectroscopic analysis of more than 12 000 individual ...stars in the globular cluster NGC 6397. Methods. The stars are deblended with a point spread function fitting technique, using a photometric reference catalogue from HST as prior, including relative positions and brightnesses. This catalogue is also used for a first analysis of the extracted spectra, followed by an automatic in-depth analysis via a full-spectrum fitting method based on a large grid of PHOENIX spectra. Results. We analysed the largest sample so far available for a single globular cluster of 18 932 spectra from 12 307 stars in NGC 6397. We derived a mean radial velocity of vrad = 17.84 ± 0.07 km s-1 and a mean metallicity of Fe/H = −2.120 ± 0.002, with the latter seemingly varying with temperature for stars on the red giant branch (RGB). We determine Teff and Fe/H from the spectra, and log g from HST photometry. This is the first very comprehensive Hertzsprung-Russell diagram (HRD) for a globular cluster based on the analysis of several thousands of stellar spectra, ranging from the main sequence to the tip of the RGB. Furthermore, two interesting objects were identified; one is a post-AGB star and the other is a possible millisecond-pulsar companion.
Aims. We demonstrate the high multiplex advantage of crowded field 3D spectroscopy with the new integral field spectrograph MUSE by means of a spectroscopic analysis of more than 12000 individual ...stars in the globular cluster NGC 6397. Methods. The stars are deblended with a point spread function fitting technique, using a photometric reference catalogue from HST as prior, including relative positions and brightnesses. This catalogue is also used for a first analysis of the extracted spectra, followed by an automatic in-depth analysis via a full-spectrum fitting method based on a large grid of PHOENIX spectra. Results. We analysed the largest sample so far available for a single globular cluster of 18 932 spectra from 12 307 stars in NGC 6397. We derived a mean radial velocity of v sub(rad)= 17.84 + or - 0.07kms super(-1) and a mean metallicity of Fe/H = -2.120 + or - 0.002, with the latter seemingly varying with temperature for stars on the red giant branch (RGB). We determine T sub(eff) and Fe/H from the spectra, and logg from HST photometry. This is the first very comprehensive Hertzsprung-Russell diagram (HRD) for a globular cluster based on the analysis of several thousands of stellar spectra, ranging from the main sequence to the tip of the RGB. Furthermore, two interesting objects were identified; one is a post-AGB star and the other is a possible millisecond-pulsar companion.
Aims. We demonstrate the high multiplex advantage of crowded field 3D spectroscopy using the new integral field spectrograph MUSE by means of a spectroscopic analysis of more than 12,000 individual ...stars in the globular cluster NGC 6397. Methods. The stars are deblended with a PSF fitting technique, using a photometric reference catalogue from HST as prior, including relative positions and brightnesses. This catalogue is also used for a first analysis of the extracted spectra, followed by an automatic in-depth analysis using a full-spectrum fitting method based on a large grid of PHOENIX spectra. Results. With 18,932 spectra from 12,307 stars in NGC 6397 we have analysed the largest sample so far available for a single globular cluster. We derived a mean radial velocity of vrad=17.84+-0.07 km/s and a mean metallicity of Fe/H=-2.120+-0.002, with the latter seemingly varying with temperature for stars on the RGB. We determine T_eff and Fe/H from the spectra, and log g from HST photometry. This is the first very comprehensive HRD for a globular cluster based on the analysis of several thousands of stellar spectra, ranging from the main sequence to the tip of the RGB. Furthermore, two interesting objects were identified with one being a post-AGB star and the other a possible millisecond-pulsar companion.
Abstract
SARS-CoV-2 variants of concern have continuously evolved and may erode vaccine induced immunity. In this observational cohort study, we determine the risk of breakthrough infection in a ...fully vaccinated cohort. SARS-CoV-2 anti-spike IgG levels were measured before first SARS-CoV-2 vaccination and at day 21–28, 90 and 180, as well as after booster vaccination. Breakthrough infections were captured through the Danish National Microbiology database. incidence rate ratio (IRR) for breakthrough infection at time-updated anti-spike IgG levels was determined using Poisson regression. Among 6076 participants, 127 and 364 breakthrough infections due to Delta and Omicron variants were observed. IRR was 0.29 (95% CI 0.15–0.56) for breakthrough infection with the Delta variant, comparing the highest and lowest quintiles of anti-spike IgG. For Omicron, no significant differences in IRR were observed. These results suggest that quantitative level of anti-spike IgG have limited impact on the risk of breakthrough infection with Omicron.
Genetic tools form the basis for the study of molecular mechanisms. Despite many recent advances in the field of genetic engineering in bacteria, genetic toolsets remain scarce for non-model ...organisms, such as the obligatory human pathogen Streptococcus pyogenes. To overcome this limitation and enable the straightforward investigation of gene functions in S. pyogenes , we have developed a comprehensive genetic toolset. By adapting and combining different tools previously applied in other Gram-positive bacteria, we have created new replicative and integrative plasmids for gene expression and genetic manipulation, constitutive and inducible promoters as well as fluorescence reporters for S. pyogenes . The new replicative plasmids feature low- and high-copy replicons combined with different resistance cassettes and a standardized multiple cloning site for rapid cloning procedures. We designed site-specific integrative plasmids and verified their integration by nanopore sequencing. To minimize the effect of plasmid integration on bacterial physiology, we screened publicly available RNA-sequencing datasets for transcriptionally silent sites. We validated this approach by designing the integrative plasmid pSpy0K6 targeting the transcriptionally silent gene SPy_1078 . Analysis of the activity of different constitutive promoters indicated a wide variety of strengths, with the lactococcal promoter P 23 showing the strongest activity and the synthetic promoter P xylS2 showing the weakest activity. Further, we assessed the functionality of three inducible regulatory elements including a zinc- and an IPTG-inducible promoter as well as an erythromycin-inducible riboswitch that showed low-to-no background expression and high inducibility. Additionally, we demonstrated the applicability of two codon-optimized fluorescent proteins, mNeongreen and mKate2, as reporters in S. pyogenes . We therefore adapted the chemically defined medium called RPMI4Spy that showed reduced autofluorescence and enabled efficient signal detection in plate reader assays and fluorescence microscopy. Finally, we developed a plasmid-based system for genome engineering in S. pyogenes featuring the counterselection marker pheS *, which enabled the scarless deletion of the sagB gene. This new toolbox simplifies previously laborious genetic manipulation procedures and lays the foundation for new methodologies to study gene functions in S. pyogenes, leading to a better understanding of its virulence mechanisms and physiology.
Hepatitis C virus (HCV) infection is prevalent among people in prison and prisons could therefore represent a unique opportunity to test risk groups for HCV. The aim of this sero-epidemiological ...study was to determine the incidence and prevalence of HCV infection and the corresponding risk factors in Danish prisons. Participants, recruited from eight Danish prisons, were tested for HCV using dried blood spots and filled out a questionaire with demographic data and risk factors for HCV infection. In total, 76.9% (801/1041) of all eligible prisoners consented to participate. The prevalence of HCV RNA positive prisoners was 4.2% (34/801) and the in-prison incidence rate was 0.7-1.0 per 100PY overall and 18-24/100PY among PWIDs. Infected prisoners were older than the overall population with a mean age of 42 years and only 17.6% (6/34) were younger than 35 years. The prevalence of PWID was 8.5% (68/801) and only 3% (2/68) of PWID were younger than 25 years. Among the PWID, 85.3% (58/68) had ever received opioid substitution therapy (OST) and 47.1% (32/68) were currently receiving OST. Risk factors associated with HCV infection were intravenous drug use, age ≥ 40 years, and being incarcerated ≥ 10 years. In conclusion, the prevalence of PWID in Danish prisons is low, possibly reflecting a decrease in injecting among the younger generation. This together with OST coverage could explain the low prevalence of HCV infection. However among PWIDs in prison the incidence remains high, suggesting a need for improved HCV prevention in prison.
Cholecystokinin-expressing interneurons (CCKIs) are hypothesized to shape pyramidal cell-firing patterns and regulate network oscillations and related network state transitions. To directly probe ...their role in the CA1 region, we silenced their activity using optogenetic and chemogenetic tools in mice. Opto-tagged CCKIs revealed a heterogeneous population, and their optogenetic silencing triggered wide disinhibitory network changes affecting both pyramidal cells and other interneurons. CCKI silencing enhanced pyramidal cell burst firing and altered the temporal coding of place cells: theta phase precession was disrupted, whereas sequence reactivation was enhanced. Chemogenetic CCKI silencing did not alter the acquisition of spatial reference memories on the Morris water maze but enhanced the recall of contextual fear memories and enabled selective recall when similar environments were tested. This work suggests the key involvement of CCKIs in the control of place-cell temporal coding and the formation of contextual memories.
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•CCKI silencing triggers broad disinhibitory network effects and enhances burst firing•CCKIs are involved in theta phase precession-mediated temporal coding•Disruption of theta phase precession did not prevent subsequent sleep replay•CCKI activity regulates the magnitude of contextual fear-memory recall
Rangel Guerrero et al. silenced the activity of CA1 Cholecystokinin-expressing interneurons. Optogenetic silencing disrupted the theta oscillation-related temporal coding of place cells during exploration while sleep replay was enhanced. Chemogenetic silencing enhanced the recall of contextual fear memories the next day and enabled the differentiation of similar environments.
IntroductionToday, it is widespread practice to postpone frozen embryo transfer (FET) in a modified natural cycle (mNC) for at least one menstrual cycle after oocyte retrieval and failed fresh embryo ...transfer or freeze-all. The rationale behind this practice is the concern that suboptimal ovarian, endometrial or endocrinological conditions following ovarian stimulation may have a negative impact on endometrial receptivity and implantation. However, two recent systematic reviews and meta-analyses based on retrospective data did not support this practice. As unnecessary delay in time to transfer and pregnancy should be avoided, the aim of this study is to investigate if immediate single blastocyst transfer in mNC-FET is non-inferior to standard postponed single blastocyst transfer in mNC-FET in terms of live birth rate.Methods and analysisMulticentre randomised controlled non-blinded trial including 464 normo-ovulatory women aged 18–40 years undergoing single blastocyst mNC-FET after a failed fresh or freeze-all cycle. Participants are randomised 1:1 to either FET in the first menstrual cycle following the stimulated cycle (immediate FET) or FET in the second or subsequent cycle following the stimulated cycle (postponed FET). The study is designed as a non-inferiority trial and primary analyses will be performed as intention to treat and per protocol.Ethics and disseminationEthical approval has been granted by the Scientific Ethical Committee of the Capital Region of Denmark (J-nr.: H-19086300). Data will be handled according to Danish law on personal data protection in accordance with the general data protection regulation. Participants will complete written consent forms regarding participation in the study and storage of blood samples in a biobank for future research. The study will be monitored by a Good Clinical Practice (GCP)-trained study nurse not otherwise involved in the study. The results of this study will be disseminated by publication in international peer-reviewed scientific journals.Trial registration numberNCT04748874; Pre-results.
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