Established bottom-up approaches for the characterization of nucleic acids (NAs) rely on the strand-cleavage activity of nucleotide-specific endonucleases to generate smaller oligonucleotides ...amenable to gas-phase sequencing. The complexity of these hydrolytic mixtures calls for the utilization of a front-end separation to facilitate full mass spectrometric (MS) characterization. This report explored the merits of microfluidic capillary zone electrophoresis (CZE) as a possible alternative to common liquid chromatography techniques. An oligonucleotide ladder was initially employed to investigate the roles of fundamental analyte features and experimental parameters in determining the outcome of CZE-MS analyses. The results demonstrated the ability to fully resolve the various rungs into discrete electrophoretic peaks with full-width half-height (FWHH) resolution that was visibly affected by the overall amount of material injected into the system. Analogous results were obtained from a digestion mixture prepared by treating yeast tRNAPhe (75 nt) with RNase T1, which provided several well-resolved peaks in spite of the increasing sample heterogeneity. The regular shapes of such peaks, however, belied the fact that most of them contained sets of comigrating species, as shown by the corresponding MS spectra. Even though it was not possible to segregate each species into an individual electrophoretic peak, the analysis still proved capable of unambiguously identifying a total of 29 hydrolytic products, which were sufficient to cover 96% of the tRNAPhe’s sequence. Their masses accurately reflected the presence of modified nucleotides characteristic of this type of substrate. The analysis of a digestion mixture obtained from the 364 nt HIV-1 5′-UTR proved to be more challenging. The electropherogram displayed fewer well-resolved peaks and significantly greater incidence of product comigration. In this case, fractionating the highly heterogeneous mixture into discrete bands helped reduce signal suppression and detection bias. As a result, the corresponding MS data enabled the assignment of 248 products out of the possible 513 predicted from the 5′-UTR sequence, which afforded 100% sequence coverage. This figure represented a significant improvement over the 36 total products identified earlier under suboptimal conditions, which afforded only 57% coverage, or the 83 observed by direct infusion nanospray-MS (72%). These results provided a measure of the excellent potential of the technique to support the bottom-up characterization of progressively larger NA samples, such as putative NA therapeutics and mRNA vaccines.
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IJS, KILJ, NUK, PNG, UL, UM, UPUK
Physical habitat requirements of fish vary with species and life-stage. Water depth and velocity have been widely suggested as strong abiotic controls on juvenile Atlantic salmon (Salmo salar L.) ...habitat. The quality of hydraulic habitat is the result of complex interactions between stream flow dynamics and stream bed morphology. In this paper we investigated the seasonal effects of flow regime and bed morphology on the availability of habitat for salmon fry over a 40-year period. The study was based in an upland Scottish river, the Girnock Burn, a long-term salmon monitoring site. We compared six sites with contrasting bed morphologies, representing the range of in-stream habitats used by salmon fry. For each site, detailed topographic surveys and daily specific discharge time series were used in a 2D hydraulic model to obtain spatially distributed water depths and mean column velocities, under different flow conditions. Normalised fish density was used as a metric to characterise potential seasonal habitat quality. This was predicted by combining previously-derived seasonal General Additive Models with 2D hydraulic models. Our results showed that sites can provide a portion of suitable habitat under a wide range of flow conditions. Habitat quality was more stable in autumn than in spring and summer using bench-marked comparisons. Marked inter-site differences occurred in spring and summer, while in autumn differences between sites were relatively small. Habitat quality time series showed consistent temporal patterns between the sites and no clear long-term trends were identified. Overall, dry-weather with low flows interspersed with high streamflow events represented the most unfavourable conditions for salmon fry with summer being the most sensitive period. Although the Girnock provides fry habitat that is generally resilient to flow variability, our results suggest that, under extremely variable and warm summer flows, mitigating strategies might be needed in order to preserve system productivity.
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•Habitat for salmon fry is strongly influenced by flow regime and bed morphology.•There is a clear seasonal effect on fish requirements and flow regime.•Dry summers provide the most unfavourable and critical habitat conditions.•No significant and consistent long-term trends in habitat quality occurred.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The aim of this study was to assess the long-term dynamics and factors associated with the serological response against the severe acute respiratory syndrome coronavirus 2 after primary infection. A ...prospective longitudinal study was conducted with monthly serological follow-up during the first 4 months, and then at 6, 8, and 10 months after the disease onset of all recovered adult in- and outpatients with coronavirus disease 2019 (COVID-19) attending Udine Hospital (Italy) during the first wave (from March to May 2020). A total of 546 individuals were included (289 female, mean age 53.1 years), mostly with mild COVID-19 (370, 68.3%). Patients were followed for a median of 302 days (interquartile range, 186 to 311). The overall seroconversion rate within 2 months was 32% for IgM and 90% for IgG. Seroreversion was observed in 90% of patients for IgM at 4 months and in 47% for IgG at 10 months. Older age, number of symptoms at acute onset, and severity of acute COVID-19 were all independent predictors of long-term immunity both for IgM (β, linear regression coefficient, 1.10, P = 0.001; β 5.15 P = 0.014; β 43.84 P = 0.021, respectively) and for IgG (β 1.43 P < 0.001; β 10.46 P < 0.001; β 46.79 P < 0.001, respectively), whereas the initial IgG peak was associated only with IgG duration (β 1.12, P < 0.001). IgM antibodies disappeared at 4 months, and IgG antibodies declined in about half of patients 10 months after acute COVID-19. These effects varied depending on the intensity of the initial antibody response, age, and burden of acute COVID-19.
Introduction
The most serious COVID‐19 deriving from severe acute respiratory syndrome coronavirus 2 causes a cytokine release storm and it is associated with worse outcomes. In COVID‐19 patients, ...interleukin‐6 (IL‐6) levels are significantly elevated. Blocking IL‐6 preliminarily resulted in the improvement of this hyperinflammatory state. It is unknown which patients could require higher doses of tocilizumab to get out of the cytokine storm.
Materials and Methods
Twenty‐four patients affected by COVID‐19 pneumonia were included. All the patients underwent tocilizumab 8 mg/kg intravenously and were tested for serum IL‐6 24 to 48 hours before and 12 to 48 hours after tocilizumab infusion. Comparisons between survivors and nonsurvivors were performed.
Results
Eighteen patients were discharged, while six patients died, with no clinical or laboratory differences between the two groups at baseline. IL‐6 was not different at baseline (P = .41), while 24 to 48 hours post‐tocilizumab IL‐6 serum levels were significantly higher in nonsurvivors than in survivors (2398.5 430.5‐9372 vs 290.5 58.5‐1305.5 pg/mL, P = .022). Serum IL‐6 post‐tocilizumab showed a good predictive ability to discriminate survivors from nonsurvivors (area under the curve, 0.815; 95% confidence interval, 0.63‐0.99, P = .02).
Conclusion
Repeated measurement of the serum level of IL‐6 early after tocilizumab may distinguish nonsurvivors from survivors and support the choice of deeper targeting IL‐6 in COVID‐19 pneumonia.
Highlights
IL‐6 could be a useful biomarker for severity of COVID‐19.
The highest burden of inflammation may be revealed by IL‐6 levels after tocilizumab.
IL‐6 levels before and after tocilizumab may indicate how to improve IL‐6 targeting therapy in COVID‐19.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
In this review, we summarise new insights into diagnostic approaches and treatment strategies for coronary artery disease (CAD) in patients with diabetes mellitus (DM). Despite the improvements in ...therapy, the clinical management of DM patients remains challenging as they develop more extensive CAD at a younger age and consistently have worse clinical outcomes than non-DM patients. Current diagnostic modalities as well as revascularisation treatments mainly focus on ischemic lesions. However, the impact of plaque morphology and composition are emerging as strong predictors of adverse cardiac events even in the absence of identified ischemia. In particular, the presence of vulnerable plaques such as thin-cap fibroatheroma (TCFA) lesions has been identified as a very strong predictor of future adverse events. This emphasises the need for an approach combining both functional and morphological methods in the assessment of lesions. In particular, optical coherence tomography (OCT) has proven to be a valuable asset by truly identifying TCFAs. New treatment strategies should consist of individualised and advanced medical regimens and may evolve towards plaque sealing through percutaneous treatment.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Objectives
Bullying victimization increases the risk of displaying internalized and externalized symptoms and alexithymia traits among adolescents. The aim of this study is to examine the mediating ...role of Alexithymia in the relationships between bullying victimization and internalized and externalized symptoms.
Methods
A total of 1092 students and their teachers (
N
= 67) of 4th to 6th grade anonymously completed a series of measures about experiences of bullying victimization, internalizing and externalizing symptoms, and alexithymia.
Results
Our data suggest that three types of bullying victimization (verbal, physical and social) predicted internalized, externalized and alexithimic symptoms. Furthermore, alexithymia partially mediated the positive effect of all the three types of bullying victimization on both internalizing and externalizing symptoms.
Conclusions
This study provided empirical data about the mediating role of alexithimia in exacerbation of internalized and externalized symptoms among adolescents and pre-adolescents exposed to three types of bullying victimization.
Cosmological scalar perturbation theory studied in the Newtonian gauge depends on two potentials Φ and Ψ . In general relativity (GR), they must coincide ( Φ = Ψ ) in the absence of anisotropic ...stresses sourced by the energy-momentum tensor. On the other hand, it is widely accepted in the literature that potential deviations from GR can be parametrized by Φ ≠ Ψ . The latter feature is therefore present in both GR cosmologies equipped with shear viscous fluids or modified gravity. We study the evolution of scalar matter density perturbations using the redshift-space-distortion-based f ( z ) σ 8 ( z ) data as a tool to differentiate and characterize the imprints of both scenarios. We show that in the f ( z ) σ 8 ( z ) evolution both scenarios yield to completely different imprints in comparison to the standard cosmology. While the current available data are not sensitive to distinguish modified gravity from viscous shear cosmologies, future precise data can be used to break this indistinguishability.
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CMK, CTK, FMFMET, IJS, NUK, PNG, UM
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