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Member States use the funds provided by the Recovery and Resilience Facility to implement ambitious reforms and investment to make their economies and societies more sustainable, resilient and prepared for the green and digital transitions. The Scoreboard gives an overview of progress in implementing the Facility and the national recovery and resilience plans. This will be followed by an ‘ex post evaluation’ in 2028, once the measures included in the recovery plans are fully implemented. The RRF Regulation requires that the Commission provides the European Parliament, the Council, the European Economic and Social Committee and the Committee of the Regions with a mid-term evaluation on the implementation of the Recovery and Resilience Facility.
The numbers on each image indicate how many worms of the examined ones displayed the indicated phenotype. (Right panels) The gonad of the same animals in the Left panels to indicate the similar developmental stage. (A) Differential interference contrast (DIC) images showing L4 worms recovered from 4-d–starved L1 worms.
The transcript level of unc-31 was increased in mir-71(lf) worms, compared with that of wild-type controls that were normalized to the value of 1. MiR-71 represses the expression of age-1 and unc-31 through the actions on their 3′UTR, but miR-71 is not required for arresting M cell division during L1 diapause. (B) The severely reduced survival rate of the mir-71(lf) mutant was suppressed by a null allele of unc-31(e928). The effect observed in ain-1(lf) mutants is likely the consequence of the combined effects of attenuating functions of these individual miRNAs. Previous studies indicate that the InsR pathway plays a dominant role in regulating L1 starvation survival and that reducing the activity of the insulin receptor daf-2, the PI3Kinase age-1, or the upstream regulator unc-31 results in increased L1 starvation survival rate (2, 3).

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L1 starvation assay was adapted from a previously described protocol (3). Worms strains were grown and maintained at 20 °C as described (29). This result is consistent with the observation that miR-71 is specifically required for the starvation-induced stress response (Fig. S5). For example, we observed a robust retarded mutant phenotype in the vulval lineage but did not see obvious defects in seam cell differentiation or alae formation. It seems plausible that miRNAs that control developmental timing are also involved in regulating the metabolic rate through repressing the InsR pathway activity.

• However, the mechanisms that coordinate the long-term survival, overall developmental arrest, and reinitiation remain to be investigated.
• (H) Fluorescence and DIC images showing that a lin-42 3′UTR reporter was repressed in mir-71(+) worms (2/2 transgenic lines) and prominently derepressed in mir-71(−) worms (2/2 transgenic lines).
• These results compelled us to examine specific interactions between individual miRNAs and their targets to gain mechanistic insights.
• (H and I) Fluorescence images (H) and statistical data (I) showing that the M cell diveded in fed animals but remained undivided in 4-, 7-, or 11-d–starved L1 wild-type and mir-71(lf) worms.
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• To determine the functional relationship of miR-71 with LIN-42 and LIT-1, mir-71(lf); lin-42(lf) L1 worms were starved for 4 d and recovered on lit-1(RNAi) plates.
• To compare the survival rates between strains, we simulated the survival rate of each genotype to 100 arbitrary “individual worms” and performed the log-rank test in Graphpad Prism 4.

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Furthermore, a recent study suggests that the expression of certain miRNAs is differentially regulated by starvation-induced dauer diapause (15). Consistent with these ideas, several recent lines of evidence suggest that miRNA let-7 revery play login and the heterochronic genes lin-42 and hbl-1 are required to regulate the starvation-induced dauer diapause (10–12) and that a number of miRNAs including lin-4 and mir-71 are involved in regulating life span (13, 14). Furthermore, worms that are long-lived due to dietary restriction or decreased mitochondrial respiratory rates are short-lived during L1 diapause, suggesting that the mechanisms controlling L1 starvation survival are different at least in some aspects from those controlling aging (3).

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Biomass recovery was similar across growth strategies, suggesting that growth-related differences play a minimal role in short-term recovery; however, early regrowth was characterised by contrasting trait shifts. Solidago canadensis exhibited high tolerance to heat and drought, with early biomass and trait recovery, indicating potential for dominance under climate extremes. Biomass fully recovered within one month in both growth strategies, but leaf traits showed transient shifts, over-recovery in SLA and under-recovery in LDMC, likely reflecting production of new leaf tissues. Please try again in a few minutes.

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We next examined the relationship between miR-71 and UNC-31, which functions upstream of AGE-1 during L1 diapause by regulating calcium-regulated dense-core vesicle fusion and the release of an insulin-like ligand (3). We identified 10 miRNA mutants that showed reduced survival rates with a stringent standard, as well as a few miRNA mutants with slightly increased survival rates (Table S1, Fig. 1D, and Fig. S1B). 1A because the ain-1 mutations reduce, but do not eliminate, miRISC functions. The overall effect of miRNAs on L1 starvation survival is expected to be significantly stronger than that reflected by the data in Fig.

L1 Starvation Survival Assay and Statistical Analysis.

We thus asked whether miR-71 was required for the reinitiation of developmental programs during the recovery phase after L1 starvation. These results suggest that miR-71 regulates the expression of unc-31 and age-1 through their 3′UTRs. Note that there are extra GFP-positive cells (red arrows) in mir-71(lf) mutants.

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The roles of InsRs have also been implicated in arresting the cell cycle in germ cells and a portion of somatic cells during L1 diapause (2, 4). Contributed new reagents/analytic tools; X.Z., R.Z., and M.H. We recommend that incorporating trait-based recovery dynamics is essential for predicting ecosystem stability under compound climate extremes.
On the other hand, the role of a particular miRNA (miR-71) is executed by repressing the expression of many genes in multiple pathways. On one hand, we showed that deletions of a good number of miRNAs have varying impacts on the L1 diapause survival rate, although they may effect the rate through different mechanisms. Instead, many specific physiological functions, such as the starvation-induced stress response, are regulated by a miRNA-target network, often involving multiple miRNAs and a large number of their targets. We found that the known developmental timing genes, hbl-1, lin-42, and lit-1, were at the top of the list (TargetScan). To understand how miR-71 affects VPC division, we searched its predicted targets for potential genes involved in regulating developmental timing. These results indicate that miR-71 plays a significant role in larval development of animals recovering from L1 diapause and likely does so by regulating the expression of components of the insulin receptor/DAF-16 pathway, as well as factors acting downstream, or in parallel to, DAF-16.
Due to how apps are automatically backed up in iOS, the backup functionality of Duo Restore is always on for iOS users who have iCloud enabled and they will not see a notification indicating their information is being backed up. NACTO connects and mobilizes North American cities and transit agencies toward safe, sustainable, and accessible transportation. We encourage customers to take advantage of our self-service options to conduct business with DCSS. Elegans Genetic Center for many mutants of miRNA and other genes.
This is consistent with hbl-1 being one of the downstream targets of miR-71, although this modest effect alone is not expected to account for the vulval developmental phenotype in mir-71 mutant. In starved L1 worms, we detected only a slight increase in the mRNA level of hbl-1 in mir-71 mutants compared with that in wild type (∼10%), which may not be biologically significant. In contrast, the mir-71(lf) mutant worms recovering on hbl-1(RNAi) displayed precocious VPC divisions similar to that seen in wild type (Fig. 4E). Consistent with the observation described above, the 4-d–starved mir-71(lf) mutants recovering on the RNAi control plates displayed the highly penetrant retarded defect in VPC division. If this were true, the starved mir-71(lf); daf-16(lf) double-mutant worms should show a slow growth phenotype similar to that of daf-16(lf) worms, but no specific VPC timing defect. (H) Fluorescence and DIC images showing that a lin-42 3′UTR reporter was repressed in mir-71(+) worms (2/2 transgenic lines) and prominently derepressed in mir-71(−) worms (2/2 transgenic lines).

• This is consistent with the previous reports that AIN-1 and AIN-2 are functional homologs with overlapping biochemical roles (16, 17).
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• Such lagged trait recovery, combined with rapid invasive recovery, suggests potential for longer-term shifts in grassland composition and function.
• The RRF Regulation requires that the Commission provides the European Parliament, the Council, the European Economic and Social Committee and the Committee of the Regions with a mid-term evaluation on the implementation of the Recovery and Resilience Facility.
• (D) A representative chart of the L1 starvation survival rates of different miRNA mutants.
• Biomass fully recovered within one month in both growth strategies, but leaf traits showed transient shifts, over-recovery in SLA and under-recovery in LDMC, likely reflecting production of new leaf tissues.

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Previous studies showed that the release of postdocking calcium-regulated dense-core vesicles, the insulin receptor (InsR) pathway, the AMPK pathway, and protein chaperones are required for the long-term survival of starved L1 worms (2–4). Unlike dauer diapause, L1 diapause is not accompanied by life cycle changes and has not been shown to require certain signaling pathways that control the formation of dauer diapause such as TGF-β signaling (daf-1, daf-7) and nuclear hormone receptor (daf-12) (2, 3). The coordinated entrance into developmental arrest, long-term survival, and the reinitiation of development upon food availability are important biological processes to investigate. Different organisms have developed versatile growth arrest strategies to overcome starvation-induced metabolic and developmental problems.
(D) Fractions of worms that carry 3′UTR reporter transgene and show no GFP expression GFP(−), weak GFP expression GFP(+/−), and comparable GFP expression to mCherry GFP(+). We found that the mRNA level of UNC-31 was up-regulated by about 20% in mir-71(lf) (Fig. 3A). These results suggest that a significant portion of the miR-71 activities in L1 diapause survival may be devoted to regulating the activities of UNC-31–mediated InsR/PI3K signaling and that the rest of miR-71 activity may regulate UNC-31–independent pathways.

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CD56highCD16+/− NK cells aredescribed by NKG2A, low level of perforin, and are primarily characterized bycytokine production.16,30,31 Therefore, it is likely that the changes in the frequencies oftwo subsets of NK cells during the disease recovery are protective mechanisms toeliminate the SARS-COV2 and thereby reducing inflammation occurred in the earlystages of disease. Our data revealed that the percentages of Th1, Th2, andTh17 cells were significantly lower in patients than healthy control (Figure 3(a)–(c) and (j)–(l), P Figure 3(d)–(h) and (m)–(q), P Figure 3(f), (h), (o), and (q), P Figure3(d), (e),(g), (m), (n), and (p)). The number of CD56low CD16+ NK cells in patients wassignificantly increased compared to healthy subjects. To determine the percentages of activated T cells, exhausted T cells, Th1 cells,Th2 cells, Th17 cells, Tregs, B cells, NK cells, and monocytes in peripheralblood of COVID-19 patients (the first day and 10 days of initiation oftherapeutic approaches) and healthy subjects, PBMCs were stained with differentmonoclonal antibodies or matched to isotype control IgG for 30 min at4○C. These changes may play afundamental role in reducing disease severity through regulating cytokineproductions involved in the inflammation and functions of various immune cells.Nevertheless, larger and more multicenter studies are needed to validate theseconclusions. A limitation of the study was the lack of determinationof immune system differences between alive and dead patients with COVID-19 during arecovery period.
Moreover, Qinet al. indicated that suppressor and helper T cell percentages were lower inpatients than normal group. In the next step, the adaptive immune system of COVID-19 subjects was studied after 1and 10 days of initiation of therapeutic methods. In an attempt to discover the frequency of other cells of innateimmunity, the number of monocytes was also assessed. As shown inFigure 4(a)–(d),statistically significant reduction in the levels of pro-inflammatory cytokines(IL-1α, IL-1β, IL-6, and TNF-α) in patients were observed during a recovery,with the exception of IL-1β level (P Figure 4(e),P Figure 4(f)). Having considered that severe COVID-19 is largely related to a cytokine storm,cytokine profiles of COVID patients were assessed during a recovery.
Having considered that innate immunity provides the early line of defense againstviral infections, some innate immune cells were studied in the course of 10 daysafter initiation of treatment, which is almost a period that the disease isdeteriorated and may result in death or recovery from COVID-19.27,28 Our datashowed that CD56lowCD16+ NK cell number was significantlylower in the early stage of recovery than the late stage of recovery; however itsfrequency was noticeably increased compared to healthy subjects. Moreover, the authors have shown that CD4+and CD8+ T cell numbers were notably decreased.17 In agreement with previous study, our data revealed that the frequencies ofTh cells (Th1, Th2, and Th17 cells) in patients were significantly lower in theearly recovery stage than the late recovery stage and healthy individuals. B cells showed an increasedpercentage in patients compared to healthy subjects, while this increase wassignificantly reduced in the late stage of recovery (Figure 3(i) and (r),P Open in a new tabThe percentages of adaptive immune cells in COVID-19 and healthyindividuals. Its frequency wassignificantly higher in the late recovery stage than early recovery stage (Figure 2(a) and (c),P highCD16+/− NK cells in the early stage of recovery was significantlyincreased in comparison with the late stage of recovery and healthy individuals(Figure 2(a) and(d),P Figure2(b) and (e),P Open in a new tabThe frequencies of innate immune cells in COVID-19 and control subjects.The percentages of CD56low CD16+ NK cells,CD56high CD16+/− NK cells, and monocytes werestudied by flow cytometry (a and b) and then analyzed (c–e).

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• This is an analytical observational (case-control) study performed on 57 patientswith COVID-19, who were referred to a COVID-19 center, Isfahan, Iran from March2020 to April 2020, and 40 healthy individuals without any the signs andsymptoms of acute respiratory infections and other health problems affected theimmune system.
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• The levels of erythrocyte sediment rate (ESR) and C-reactive protein (CRP) ofCOVID-19 patients were measured using the erythrocyte sedimentation rate (ESR)analyzer (Parsian Teb, Iran) and Mindray BS-800 automated biochemistry analyzer(Shenzhen Mindray Bio-Medical Electronics, China), respectively.

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The RRF funding amounts shown for measures are based on the initial cost estimates provided by the Member States included in the recovery and resilience plans and may as such differ from finalized Commission financial reports, which follow eventual project implementation. The map exclusively serves information purposes and is not an exhaustive database of projects supported by the Recovery and Resilience Facility. Improving access to and the quality of general, vocational, and higher education; focusing on digital education, early childhood education and care; supporting youth employment. The funding amounts shown reflect the initial cost estimates included in the national recovery and resilience plans. It is not an exhaustive database of projects supported by the Facility and will be regularly updated as the implementation progresses.

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The plasma levels ofIL-1a, IL-1β, IL-6, TNF-α, and IL-10 were significantly higher in COVID-19 patientsthan the control group. The depicted results arerepresentative of 40 independent experiments for control group, 57independent experiments for COVID-19 patients at the first day oftreatment, and 51 independent experiments for COVID-19 patients in10 days of treatment. Data were analyzed by GraphPad Prism 6 (GraphPad Software, USA) and are expressedas the mean standard error of the mean (SEM) and mean ± standard deviation (SD).The normal distribution of data was determined by Kolmogrov–Smirnov test. The levels of erythrocyte sediment rate (ESR) and C-reactive protein (CRP) ofCOVID-19 patients were measured using the erythrocyte sedimentation rate (ESR)analyzer (Parsian Teb, Iran) and Mindray BS-800 automated biochemistry analyzer(Shenzhen Mindray Bio-Medical Electronics, China), respectively.
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This studyinvestigated how the immune system changes were related to disease severity inCOVID-19 patients. Moreover, other data indicated thatthe levels of these cytokines were reduced during the disease recovery. PBMCs were isolated from healthy subjects and COVID-19patients and then stained with different monoclonal antibodies. To determine the situations of humoral and cellular immunity in patients withCOVID-19, the frequencies of Th1, Th2, Th17, Treg, activated CD4+T cells,activated CD8+ T cells, exhausted CD4+ T cells, exhausted CD8+ T cells, and Bcells in COVID-19 patients were investigated after 1 and 10 days of initiationof therapeutic methods. Correlations of lymphocyte numbers with the value of ESR and numbers ofTh2 cells and monocytes in COVID-19 patients. Some patients hadfatigue, mild shortness of breath, myalgia, loss of weight, smell, and taste inthe late recovery stage.

Map of projects supported by the Recovery and Resilience Facility

The CD3+ cell population was also determined using the gating oflymphocyte population and was then used to measure the percentages of B cells(CD3− CD19+ CD22+ cells), exhausted CD4+ T cells (CD3+ CD4+ PD-1+ cells),exhausted CD8+ T cells (CD3+ CD8+ PD-1+ cells), CD56lowCD16+ NK cells (CD3− CD56lowCD16+ cells), and CD56high CD16+/− NK cells(CD3−CD56high CD16+/− cells). Afterwards, the lymphocyte population was gatedto assess the frequencies of the CD4+ cells which were used to determine thepercentages of Th1 cells (CD4+ T-bet+ IFN-γ+ cells), Th2 cells (CD4+ IL-4+GATA3+ cells), Th17 cells (CD4+ IL-17α+ RORγt+ cells), Tregs (CD4+CD127low FoxP3+ cells), and activated CD4+ T cells (CD4+ CD25+CD69+ cells). At the first day (the early recovery stage) and 10 days of initiation oftherapeutic methods (the late recovery stage), heparinized blood samples (5 ml)were obtained from patients. All patients had pulmonary involvement and were not on treatment withdrugs influencing the immune system and antibodies production (i.e. steroids,sulfasalazine, phenytoin, and antimalarial drugs) prior to study initiation.

COVID-19 Response

Other results of the currentstudy revealed that the percentage of another subset of NK cells(CD56highCD16+/− NK cells) was significantly increased atthe first day of recovery. TheCD56lowCD16+ NK cells have high expression levels ofkiller inhibitory receptors, the maturation marker (CD57), and natural andantibody-dependent cellular cytotoxicity which is mediated by releasing high levelsof perforin and enhanced killing.16,30,31 These findings suggest thatthe reduced number of CD56lowCD16+ NK cells may contribute todisease susceptibility in the early stages of disease. COVID-19, as a pandemic disease, is responsible for considerable mortality and morbidity.25 Immune system functions have fundamental role in the pathogenesis and outcomeof disease.26 Therefore, the current study focused on determining how immune system changesduring a recovery were correlated to disease severity. Thepercentages of Th1, Th2, Th17, Tregs, exhausted CD4+ T cells, exhaustedCD8+ T cells, activated CD4+ T cells, activated CD8+ T cells, and Bcells were assessed using flow cytometry (a–i) and then analyzed (j–r).The depicted results are representative of 57 independent experimentsfor COVID-19 patients at the first day of treatment, 51 independentexperiments for COVID-19 patients in 10 days of treatment, and 40independent experiments for healthy groups. The frequencies of innate immune cells in COVID-19 and control subjects.The percentages of CD56low CD16+ NK cells,CD56high CD16+/− NK cells, and monocytes werestudied by flow cytometry (a and b) and then analyzed (c–e).
In this study, the mean ± SD of age of patients was 67.8 ± 15.18, while it was66.01 ± 7.11 in healthy subjects. In thisstudy, CD8+ CD25+ CD69+ cells and CD14+ CD16+ CD11b+ cells were respectivelyconsidered as the activated CD8+ T cells and monocytes. To determine the immune situation of patients, theblood sampling (5 ml) from healthy subjects was also performed. This is an analytical observational (case-control) study performed on 57 patientswith COVID-19, who were referred to a COVID-19 center, Isfahan, Iran from March2020 to April 2020, and 40 healthy individuals without any the signs andsymptoms of acute respiratory infections and other health problems affected theimmune system. Although the pathogenesis of COVID-19 is not well understood yet, defects in functionand/or regulation of the immune system such as the storm of inflammatory cytokinesand lymphopenia can contribute to the intensity of pathogenic coronavirusinfections.11–13 In despite ofsome reports pointing to impacts of immune responses in the pathogenesis of COVID-19,14 the accurate roles of immune cells in developing or inhibiting the diseaseare unknown.
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• To determine the situations of humoral and cellular immunity in patients withCOVID-19, the frequencies of Th1, Th2, Th17, Treg, activated CD4+T cells,activated CD8+ T cells, exhausted CD4+ T cells, exhausted CD8+ T cells, and Bcells in COVID-19 patients were investigated after 1 and 10 days of initiationof therapeutic methods.
• The number of lymphocytes in peripheral blood of COVID-19 patients in the earlyand late stages of recovery and healthy subjects were assessed by an automatedcell counter system UF-100® (Sysmex, Kobe, Japan) within 3 h aftercollecting blood samples.
• To determine the percentages of activated T cells, exhausted T cells, Th1 cells,Th2 cells, Th17 cells, Tregs, B cells, NK cells, and monocytes in peripheralblood of COVID-19 patients (the first day and 10 days of initiation oftherapeutic approaches) and healthy subjects, PBMCs were stained with differentmonoclonal antibodies or matched to isotype control IgG for 30 min at4○C.
• The results indicated thatpatients had the reduced number of lymphocyte in comparison with healthy subjects.In line with this finding, Qin et al. declared that patients with COVID-19 had areduction in T cell number accompanied by the severity of the disease.
• To determine cytokine profiles in COVID-19 patients, the plasma samples wereobtained from whole blood (5 ml) of participants and the levels of IL-1α, IL-1β,TNF-α, IL-6, TGF-β1, and IL-10 cytokines were measured using an Enzyme-linkedimmunoasorbent assay (ELISA) kit (Karmania Pars Gene, Iran) according themanufacturer’s instructions.

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In disagreement with other reports showing increasedfrequency of B cells in the late stage of recovery,17 we observed that the percentage of this cell was decreased followingrecovery. Moreinterestingly, the percentages of exhausted CD4+ T cells and exhausted CD8+ T cellswere higher in the early stage of recovery than the late stage of recovery. Thisobservation was in contrast with previous study showing severe cases of COVID-19tend to have lower percentages of monocytes.24 This discrepancy may be attributed to disease stage which patients wereevaluated.

The resultsare representative of 57 independent experiments for COVID-19 patientsat the first day of treatment, 51 independent experiments for COVID-19patients in 10 days of treatment, and 40 independent experiments forhealthy individuals. The demographic, laboratory, and clinical characteristics of COVID-19 andhealthy subjects. Table2 depicts the demographic and other characteristics of COVID-19 andhealthy subjects. Of the 57 patients, 51 (89.48%) weredischarged from hospital and 6 (10.52%) died during the study. Antibodies used for determing the changes of the immune system ofCOVID-19 patients by flow cytometry.
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I know the email address and password. I lost my account close to 2 months ago when I deleted the game trying to fix a login issue. I was wondering if there is anyone who has not spent any money on the game lost their account. Improving the resilience, accessibility and quality of health and long-term care, including measures to advance their digitalisation; increasing the effectiveness of public administration systems. Improving social and territorial infrastructure and services, including social protection and welfare systems, the inclusion of disadvantaged groups; supporting employment and skills development; creating high-quality, stable jobs. Promoting the roll-out of very high-capacity networks, the digitalisation of public services, government processes, and businesses, in particular SMEs; developing basic and advanced digital skills; supporting digital-related R&D and the deployment of advanced technologies.

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