Analysis of retinal and choroidal microvascular changes using optical coherence tomography and optical coherence tomography angiography in patients with acute leukemia.

PURPOSE: Although leukemic retinopathy accounts for 80% of ocular complications in acute leukemia, its pathogenesis remains unclear. To evaluate changes in retinal and choroicapillaris and structural parameters in patients with acute leukemia, we analyzed the correlation between vascular perfusion metrics and laboratory parameters and assessed the changes after hematopoietic stem cell transplantation (HSCT). METHODS: Herein, 104 eyes of 52 patients aged 18 and above with acute leukemia were enrolled. 80 eyes of 40 healthy patients were recruited as control participants. All participants underwent optical coherence tomography (OCT) and OCT angiography (OCTA) at baseline. RESULTS: Patients with acute leukemia had a significantly thicker ganglion cell-inner plexiform layer (GCIPL) and lower circularity index than the control participants. Post-HSCT perfusion metrics did not differ significantly, but parafoveal thickness decreased significantly. During the active phase of acute leukemia, lower platelet levels were associated with significant GCIPL thickening and increased foveal avascular zone and perimeter. D-dimer levels positively correlated with GCIPL thickness. CONCLUSION: Patients with acute leukemia had subclinical retinal microvascular deficits on OCTA and GCIPL thickening on OCT, possibly associated with bone marrow function. GCIPL thickness may indicate acute ischemia in such patients. Further studies must elucidate their clinical and prognostic significance.

Quantifying the clusterness and trajectoriness of single-cell RNA-seq data.

Among existing computational algorithms for single-cell RNA-seq analysis, clustering and trajectory inference are two major types of analysis that are routinely applied. For a given dataset, clustering and trajectory inference can generate vastly different visualizations that lead to very different interpretations of the data. To address this issue, we propose multiple scores to quantify the "clusterness" and "trajectoriness" of single-cell RNA-seq data, in other words, whether the data looks like a collection of distinct clusters or a continuum of progression trajectory. The scores we introduce are based on pairwise distance distribution, persistent homology, vector magnitude, Ripley's K, and degrees of connectivity. Using simulated datasets, we demonstrate that the proposed scores are able to effectively differentiate between cluster-like data and trajectory-like data. Using real single-cell RNA-seq datasets, we demonstrate the scores can serve as indicators of whether clustering analysis or trajectory inference is a more appropriate choice for biological interpretation of the data.

PINK1 restrains periodontitis-induced bone loss by preventing osteoclast mitophagy impairment.

The oral colonization of periodontal pathogens onto gingival tissues establishes hypoxic microenvironment, often disrupting periodontal homeostasis in conjunction with oxidative stress. The association between reactive oxygen species (ROS) and osteolytic periodontitis have been suggested by recent studies. PTEN-induced kinase 1 (PINK1), a mitochondrial serine/threonine kinase, is an essential protein for mitochondrial quality control as it protects cells from oxidative stress by promoting degradation of damaged mitochondria through mitophagy. However, the pathophysiological roles of PINK1 in osteoclast-mediated bone loss have not been explored. Here we aimed to determine whether PINK1 plays a role in the regulation of osteoclastogenesis and alveolar bone resorption associated with periodontitis. C57BL/6 wild type (WT) and Pink1 knockout (KO) mice were subjected to ligature-induced periodontitis (LIP), and alveolar bones were evaluated by µCT-analysis and tartrate-resistant acid phosphatase (TRAP) staining. The µCT-analysis showed that bone volume fraction and travecular thickness were lower in Pink1 KO compared to WT mice. The number of TRAP-positive osteoclasts was markedly increased in the periodontal tissues of Pink1 KO mice with LIP. The genetic silencing or deletion of Pink1 promoted excessive osteoclast differentiation and bone resorption in vitro, as respectively indicated by TRAP staining and resorption pits on dentin slices. PINK1 deficiency led to mitochondrial instabilities as indicated by confocal microscopy of mitochondrial ROS, mitochondrial oxygen consumption rate (OCR) analysis, and transmission electron microscopy (TEM). Consequently, a significant increase in Ca2+-nuclear factor of activated T cells 1 (NFATc1) signaling was also found. On the other hand, restoration of mitophagy and autophagy by spermidine (SPD) treatment and the resolution of oxidative stress by N-acetyl-l-cysteine (NAC) treatment protected PINK1 deficiency-induced excessive generation of osteoclasts. Taken together, our findings demonstrate that PINK1 is essential for maintaining mitochondrial homeostasis during osteoclast differentiation. Therefore, targeting PINK1 may provide a novel therapeutic strategy for severe periodontitis with fulminant osteolysis.

Prevalence of adolescent-reported food insecurity and the determinants including coping strategies living in urban slum communities of Bangladesh during the era of COVID-19: a cross-sectional study.

BACKGROUND: As food insecurity (FI) continues to rise worldwide especially in developing countries like Bangladesh, adolescent experience of FI have received minimal attention globally. This study aimed to identify the prevalence of adolescent-reported FI and its association with individual and socio-environmental factors as well as coping strategies amongst a sample of adolescents living in urban slum areas of Bangladesh in the times of the Coronavirus 19 (COVID-19) pandemic. METHODS: A descriptive cross sectional study was conducted amongst 326 adolescents (12-18 years) living in the urban slums of Narayanganj, Dhaka from April to May, 2022. Adolescent-reported FI was assessed using a structured questionnaire adopted from Household Food Insecurity Access Scale (HFIAS). Descriptive statistics, Chi-square tests and ordinal logistic regression were used to draw inference. RESULTS: Prevalence of adolescent-reported FI was high (46.6% moderate and 29.8% severe). The likelihood of experiencing moderate or severe FI versus no/mild FI were 1.7 times (95% Confidence Interval (CI) [1.1, 2.5]) higher in younger adolescents and 5 times (95% CI [2.3, 12.7]) higher in unemployed youth. Socio-environmental factors determining the economic status of a household such as higher number of family members, only one earning family member, unemployed father, no household assets, food aid received by the family during pandemic and positive COVID-19 infection in family were associated with moderate and severe FI. Coping strategies such as a higher number of food seeking strategies (Adjusted Odds Ratio (AOR) 3.4, 95% CI [1.9, 5.9]), substance use (AOR 6.2, 95% CI [1.2, 31.7]) and stopping school (AOR 3.3, 95% CI [1.9, 5.7]) increased odds for moderate and severe FI. Stratified by drop-out of school status, an association between food seeking strategies and FI remained significant among those school-going, while there was no association among those dropping out of school. CONCLUSION: This study showed that adolescents living in urban slum communities in Bangladesh are at very high risk of FI and resort to harmful coping strategies. Our study highlights the importance of further research in adolescent reported FI and coping strategies in low to middle income countries (LMICs) and create appropriate interventions to lower FI among this group and improve their state of health and wellbeing.

Molecular mechanisms of SARS-CoV-2 resistance to nirmatrelvir.

Nirmatrelvir is a specific antiviral drug that targets the main protease (Mpro) of SARS-CoV-2 and has been approved to treat COVID-191,2. As an RNA virus characterized by high mutation rates, whether SARS-CoV-2 will develop resistance to nirmatrelvir is a question of concern. Our previous studies have shown that several mutational pathways confer resistance to nirmatrelvir, but some result in a loss of viral replicative fitness, which is then compensated for by additional alterations3. The molecular mechanisms for this observed resistance are unknown. Here we combined biochemical and structural methods to demonstrate that alterations at the substrate-binding pocket of Mpro can allow SARS-CoV-2 to develop resistance to nirmatrelvir in two distinct ways. Comprehensive studies of the structures of 14 Mpro mutants in complex with drugs or substrate revealed that alterations at the S1 and S4 subsites substantially decreased the level of inhibitor binding, whereas alterations at the S2 and S4' subsites unexpectedly increased protease activity. Both mechanisms contributed to nirmatrelvir resistance, with the latter compensating for the loss in enzymatic activity of the former, which in turn accounted for the restoration of viral replicative fitness, as observed previously3. Such a profile was also observed for ensitrelvir, another clinically relevant Mpro inhibitor. These results shed light on the mechanisms by which SARS-CoV-2 evolves to develop resistance to the current generation of protease inhibitors and provide the basis for the design of next-generation Mpro inhibitors.

Effect of Cooling Temperature on Crystalline Behavior of Polyphenylene Sulfide/Glass Fiber Composites.

Poly (phenylene sulfide) (PPS) is a super engineering plastic that has not only excellent rigidity and high chemical resistance but also excellent electrical insulation properties; therefore, it can be applied as an electronic cover or an overheating prevention component. This plastic has been extensively applied in the manufacture of capacitor housing as, in addition to being a functional and lightweight material, it has a safety feature that can block the electrical connection between the electrolyte inside and outside the capacitor. Moreover, the fabrication of PPS composites with high glass fiber (GF) content facilitates the development of lightweight and excellent future materials, which widens the scope of the application of this polymer. However, the crystallinity and mechanical properties of PPS/GF composites have been found to vary depending on the cooling temperature. Although extensive studies have been conducted on the influence of cooling temperature on the crystalline behavior of PPS-based composites, there has been limited research focused particularly on PPS/GF composites for capacitor housing applications. In this study, to apply PPS/GF composites as film capacitor housings, specimens were prepared via injection molding at different cooling temperatures to investigate the composites' tensile, flexural, and impact energy absorption properties resulting in increases in mechanical properties at high cooling mold temperature. Fracture surface analysis was also performed on the fractured specimens after the impact test to confirm the orientation of the GF and the shape of the micropores. Finally, the crystallinity of the composites increased with higher cooling temperatures due to the extended crystallization time.

Identifying dysregulated immune cell subsets following volumetric muscle loss with pseudo-time trajectories.

Volumetric muscle loss (VML) results in permanent functional deficits and remains a substantial regenerative medicine challenge. A coordinated immune response is crucial for timely myofiber regeneration, however the immune response following VML has yet to be fully characterized. Here, we leveraged dimensionality reduction and pseudo-time analysis techniques to elucidate the cellular players underlying a functional or pathological outcome as a result of subcritical injury or critical VML in the murine quadriceps, respectively. We found that critical VML resulted in a sustained presence of M2-like and CD206hiLy6Chi 'hybrid' macrophages whereas subcritical defects resolved these populations. Notably, the retained M2-like macrophages from critical VML injuries presented with aberrant cytokine production which may contribute to fibrogenesis, as indicated by their co-localization with fibroadipogenic progenitors (FAPs) in areas of collagen deposition within the defect. Furthermore, several T cell subpopulations were significantly elevated in critical VML compared to subcritical injuries. These results demonstrate a dysregulated immune response in critical VML that is unable to fully resolve the chronic inflammatory state and transition to a pro-regenerative microenvironment within the first week after injury. These data provide important insights into potential therapeutic strategies which could reduce the immune cell burden and pro-fibrotic signaling characteristic of VML.

Social support for postpartum women and associated factors including online support to reduce stress and depression amidst COVID-19: Results of an online survey in Thailand.

BACKGROUND: Social support for postpartum women helps mothers to recover from childbirth and fosters healthy infant development. However, the impacts of reduced interpersonal interactions inflicted by the COVID-19 outbreak on available social support for postpartum women have received little attention. Therefore, this study aimed to examine the levels of social support provided to postpartum women and associated factors in Thailand during the COVID-19 pandemic. METHODS: A cross-sectional study was conducted from July to October 2021 using an anonymous online questionnaire. The responses of 840 eligible women up to six months postpartum in Thailand were obtained. The maternity social support scale was used to measure social support. Multivariate logistic regression was used to analyse the factors associated with social support among postpartum women. RESULTS: About 57% of women reported to receive high support. Women in the high social support group were more likely to be married (aOR:2.70; 95% CI:1.57-4.66), have a university education or above (1.88; 1.35-2.64), have an intended pregnancy (2.06; 1.34-3.16), good health (2.01; 1.44-2.81), good sleep quality (1.62; 1.14-2.31), receive counsel from peers or family (1.56; 1.13-2.16), and use internet or social media to reduce stress and depression (1.51; 1.08-2.11). Meanwhile, women in the high social support group were significantly less likely to feed complementary foods to infants within 24 hours of completing the survey (0.28; 0.15-0.52). CONCLUSIONS: The results of this study indicated that more than half of the women reported high support and illustrated the important role played by family, peers, and professionals as well as online and remote channels in providing postpartum informational and emotional support during the pandemic. Online platforms and remote support may be considered to provide social support to postpartum women during a pandemic such as COVID-19.

Stromal STAT5-Mediated Trophic Activity Regulates Hematopoietic Niche Factors.

Signal transducer and activator of transcription 5 (STAT5a and STAT5b) are intrinsically critical for normal hematopoiesis but are also expressed in stromal cells. Here, STAT5ab knockout (KO) was generated with a variety of bone marrow hematopoietic and stromal Cre transgenic mouse strains. Vav1-Cre/+STAT5abfl/fl, the positive control for loss of multipotent hematopoietic function, surprisingly dysregulated niche factor mRNA expression, and deleted STAT5ab in CD45neg cells. Single-cell transcriptome analysis of bone marrow from Vav1-Cre/+ wild-type or Vav1-Cre/+STAT5abfl/fl mice showed hematopoietic stem cell (HSC) myeloid commitment priming. Nes+ cells were detected in both CD45neg and CD45+ clusters and deletion of STAT5ab with Nes-Cre caused hematopoietic repopulating defects. To follow up on these promiscuous Cre promoter deletions in CD45neg and CD45+ bone marrow cell populations, more stroma-specific Cre strains were generated and demonstrated a reduction in multipotent hematopoietic progenitors. Functional support for niche-supporting activity was assessed using STAT5-deficient mesenchymal stem cells (MSCs). With Lepr-Cre/+STAT5abfl/fl, niche factor mRNAs were downregulated with validation of reduced IGF-1 and CXCL12 proteins. Furthermore, advanced computational analyses revealed a key role for STAT5ab/Cish balance with Cish strongly co-expressed in MSCs and HSCs primed for differentiation. Therefore, STAT5ab-associated gene regulation supports the bone marrow microenvironment.

Antiarrhythmic Effect of Artemisinin in an Ex-vivo Model of Brugada Syndrome Induced by NS5806.

BACKGROUND AND OBJECTIVES: Brugada syndrome (BrS) is an inherited arrhythmia syndrome that presents as sudden cardiac death (SCD) without structural heart disease. One of the mechanisms of SCD has been suggested to be related to the uneven dispersion of transient outward potassium current (Ito) channels between the epicardium and endocardium, thus inducing ventricular tachyarrhythmia. Artemisinin is widely used as an antimalarial drug. Its antiarrhythmic effect, which includes suppression of Ito channels, has been previously reported. We investigated the effect of artemisinin on the suppression of electrocardiographic manifestations in a canine experimental model of BrS. METHODS: Transmural pseudo-electrocardiograms and epicardial/endocardial transmembrane action potentials (APs) were recorded from coronary-perfused canine right ventricular wedge preparations (n=8). To mimic the BrS phenotypes, acetylcholine (3 µM), calcium channel blocker verapamil (1 µM), and Ito agonist NS5806 (6-10 µM) were used. Artemisinin (100-150 µM) was then perfused to ameliorate the ventricular tachyarrhythmia in the BrS models. RESULTS: The provocation agents induced prominent J waves in all the models on the pseudo-electrocardiograms. The epicardial AP dome was attenuated. Ventricular tachyarrhythmia was induced in six out of 8 preparations. Artemisinin suppressed ventricular tachyarrhythmia in all 6 of these preparations and recovered the AP dome of the right ventricular epicardium in all preparations (n=8). J wave areas and epicardial notch indexes were also significantly decreased after artemisinin perfusion. CONCLUSIONS: Our findings suggest that artemisinin has an antiarrhythmic effect on wedge preparation models of BrS. It might work by inhibition of potassium channels including Ito channels, subsequently suppressing ventricular tachycardia/ventricular fibrillation.

Quantifying Cell-Type-Specific Differences of Single-Cell Datasets Using Uniform Manifold Approximation and Projection for Dimension Reduction and Shapley Additive exPlanations.

With rapid advances in single-cell profiling technologies, larger-scale investigations that require comparisons of multiple single-cell datasets can lead to novel findings. Specifically, quantifying cell-type-specific responses to different conditions across single-cell datasets could be useful in understanding how the difference in conditions is induced at a cellular level. In this study, we present a computational pipeline that quantifies cell-type-specific differences and identifies genes responsible for the differences. We quantify differences observed in a low-dimensional uniform manifold approximation and projection for dimension reduction space as a proxy for the difference present in the high-dimensional space and use SHapley Additive exPlanations to quantify genes driving the differences. In this study, we applied our algorithm to the Iris flower dataset, single-cell RNA sequencing dataset, and mass cytometry dataset and demonstrate that it can robustly quantify cell-type-specific differences and it can also identify genes that are responsible for the differences.

Effects of multimodal cognitive exercise program on cognitive function, bone density, blood lipid, fitness, and depression in old women with mild cognitive impairment.

The purpose of this study is to verify the effects of participation in 12-week multimodal cognitive exercise program on cognitive function, bone mineral density, blood lipids, senior functional fitness, and depression of the old women with mild cognitive impairment and osteopenia. As a result, the exercise group showed significant effects in cognitive function, depression, bone mineral density, blood lipid, lower extremity flexibility, upper extremity flexibility, dynamic balance, and body endurance. Such results prove that multimodal cognitive exercise program is an effective interventional exercise program for improving cognitive function in the old women with mild cognitive impairment.

Maternal Depression during Pregnancy and Postpartum Period among the Association of Southeast Asian Nations (ASEAN) Countries: A Scoping Review.

Identification of mothers with depression is important because untreated perinatal depression can have both short- and long-term consequences for the mother, the child, and the family. This review attempts to identify the prevalence of antenatal and postnatal depression (AD and PD, respectively) of mothers among the ASEAN member countries. A literature review was conducted using PubMed, Scopus, and the Asian Citation Index. The reviews covered publications in peer-reviewed journals written in the English language between January 2010 and December 2020. Of the 280 articles identified, a total of 37 peer-reviewed articles conducted in 8 out of 11 ASEAN member countries were included. The Edinburgh Postnatal Depression Scale (EPDS) was the most common instrument used to identify depression. This study showed the number of studies reporting the prevalence of AD was 18 in five countries. For PD, 24 studies in eight countries were included. The prevalence of AD ranged from 4.9% to 46.8%, and that of PD ranged from 4.4% to 57.7%. This first review among ASEAN countries showed very few studies conducted in lower-middle-income and substantial heterogeneity in prevalence among studies reviewed. Further research should be conducted to estimate the prevalence using a large representative sample with a validated assessment tool among the ASEAN countries.

Factors associated with postpartum depression symptoms among postpartum women in five countries during the COVID-19 pandemic: an online cross-sectional study.

BACKGROUND: This study aimed to examine factors associated with postpartum depression (PPD) symptoms during the COVID-19 pandemic among postpartum women in five countries, a subject that has not been investigated thus far. METHODS: A multi-country, cross-sectional, online survey was conducted with a convenience sample of 3,523 postpartum women in Brazil, South Korea, Taiwan, Thailand, and the United Kingdom, from July to November 2021. Sociodemographic and obstetric data, food insecurity, COVID-19 positive status, COVID-19 vaccination, infant feeding, breastfeeding belief score, and social support were investigated. PPD and social support were measured using the Edinburgh Postnatal Depression Scale and Maternal Social Support Scale, respectively. Descriptive statistics, chi-squared tests, and t-tests were used to identify associations with PPD symptoms. A binary logistic regression model was used to identify explanatory factors associated with PPD and adjusted odds ratios (OR) and 95% confidence intervals (CIs) were calculated. RESULTS: Women in Taiwan (AOR = 0.5; 95%CI 0.34, 0.73) and Thailand (AOR = 0.68; 95%CI 0.46, 0.99) had a lower risk of PPD symptoms than those in Brazil. In addition, women with planned pregnancies had a lower risk of PPD (AOR = 0.74; 95%CI 0.60, 0.91). Younger women (AOR = 1.62; 95%CI 1.05, 2.51), health problems during pregnancy, delivery, or postpartum (AOR = 1.71; 95%CI 1.42, 2.06), and no change or worse food insecurity during COVID-19 (AOR = 1.66; 95%CI 1.21, 1.27 for no change and AOR = 1.68; 95%CI 1.27, 1.23, respectively) presented a higher likelihood of having PPD. Feeding babies with expressed human milk (AOR = 1.25; 95%CI 1.03, 1.50) and/or complementary food (AOR = 1.51; 95%CI 1.17, 1.94) were associated with PPD symptoms. Women who received low (AOR = 7.74; 95%CI 5.43, 11.03) or medium support (AOR = 3.25; 95%CI 2.71, 3.88) had higher likelihoods of PPD. CONCLUSION: PPD symptoms during the pandemic were high in young women, particularly Brazilian women, with health problems in the puerperal pregnancy cycle who fed their babies expressed breast milk and/or complementary food. Low social support also impacted PPD symptoms. This study highlights the need for the professional screening for PPD and provision of virtual or personal support.

Embracing enzyme promiscuity with activity-based compressed biosensing.

The development of protease-activatable drugs and diagnostics requires identifying substrates specific to individual proteases. However, this process becomes increasingly difficult as the number of target proteases increases because most substrates are promiscuously cleaved by multiple proteases. We introduce a method-substrate libraries for compressed sensing of enzymes (SLICE)-for selecting libraries of promiscuous substrates that classify protease mixtures (1) without deconvolution of compressed signals and (2) without highly specific substrates. SLICE ranks substrate libraries using a compression score (C), which quantifies substrate orthogonality and protease coverage. This metric is predictive of classification accuracy across 140 in silico (Pearson r = 0.71) and 55 in vitro libraries (r = 0.55). Using SLICE, we select a two-substrate library to classify 28 samples containing 11 enzymes in plasma (area under the receiver operating characteristic curve [AUROC] = 0.93). We envision that SLICE will enable the selection of libraries that capture information from hundreds of enzymes using fewer substrates for applications like activity-based sensors for imaging and diagnostics.

Prevalence and factors associated with treatment and control of hypertension among adults with hypertension in Myanmar.

BACKGROUND: Due to a dearth in the number of studies conducted in low- and middle-income countries, this study aimed to identify the prevalence and determinants of the treatment and control of hypertension among patients with hypertension in Myanmar. METHODS: This community-based cross-sectional study was conducted among 410 adults who were registered for hypertensive treatment in health centers in Myitkyina Township, Kachin State, Myanmar. Multiple logistic regression was used to identify the associated factors. RESULTS: The prevalence of treatment and control of hypertension was 48.1% and 20.5%, respectively. The factors associated with treatment were age (OR=2.60 for 46-60 y and OR=2.29 for 61-70 y compared with 30-45 y), ethnicity (OR=1.87), monthly family income (OR=1.90), comorbidity (OR=2.33), knowledge (OR=2.63) and adherence to physical activity (OR=1.86). Controlled hypertension was associated with age (OR=3.03 for 46-60 y and OR=2.27 for 61-70 y compared with 30-45 y), education (OR=1.81), comorbidity (OR=1.67) and adherence to medication (OR=3.45). CONCLUSIONS: The prevalence of treated and controlled hypertension was relatively low in this study. To improve the prevalence of hypertension treatment and control in this study population, effective and culturally sensitive intervention programs under universal health coverage should be established with an emphasis on individuals with lower educational attainment and younger ages.

Associations between breastfeeding intention, breastfeeding practices and post-natal depression during the COVID-19 pandemic: A multi-country cross-sectional study.

Associations between breastfeeding intention, duration and post-natal depression (PND) have been shown in pre-COVID-19 studies. However, studies during COVID-19 have not examined the associations between breastfeeding intention, breastfeeding practices, and PND in an international sample of post-natal women, taking into consideration COVID-19 related factors. This is the first study to address this gap as both PND and breastfeeding may be affected by COVID-19, and have important long-term effects on women's and infant's health. A cross-sectional internet-based survey was conducted with 3253 post-natal women from five countries: Brazil, South Korea, Taiwan, Thailand, and the United Kingdom from July to November 2021. The results showed that women who intended to breastfeed during pregnancy had lower odds of having PND than women who did not intend to. Women who had no breastfeeding intention but actually breastfed had greater odds (AOR 1.75) of having PND than women who intended to breastfeed and actually breastfed. While there was no statistical significance in expressed breast milk feeding in multivariable logistic regression models, women who had shorter duration of breastfeeding directly on breast than they planned had greater odds (AOR 1.58) of having PND than those who breastfed longer than they planned even after adjusting for covariates including COVID-19-related variables. These findings suggested the importance of working with women on their breastfeeding intention. Tailored support is required to ensure women's breastfeeding needs are met and at the same time care for maternal mental health during and beyond the pandemic.

Multiple pathways for SARS-CoV-2 resistance to nirmatrelvir.

Nirmatrelvir, an oral antiviral targeting the 3CL protease of SARS-CoV-2, has been demonstrated to be clinically useful against COVID-19 (refs. 1,2). However, because SARS-CoV-2 has evolved to become resistant to other therapeutic modalities3-9, there is a concern that the same could occur for nirmatrelvir. Here we examined this possibility by in vitro passaging of SARS-CoV-2 in nirmatrelvir using two independent approaches, including one on a large scale. Indeed, highly resistant viruses emerged from both and their sequences showed a multitude of 3CL protease mutations. In the experiment peformed with many replicates, 53 independent viral lineages were selected with mutations observed at 23 different residues of the enzyme. Nevertheless, several common mutational pathways to nirmatrelvir resistance were preferred, with a majority of the viruses descending from T21I, P252L or T304I as precursor mutations. Construction and analysis of 13 recombinant SARS-CoV-2 clones showed that these mutations mediated only low-level resistance, whereas greater resistance required accumulation of additional mutations. E166V mutation conferred the strongest resistance (around 100-fold), but this mutation resulted in a loss of viral replicative fitness that was restored by compensatory changes such as L50F and T21I. Our findings indicate that SARS-CoV-2 resistance to nirmatrelvir does readily arise via multiple pathways in vitro, and the specific mutations observed herein form a strong foundation from which to study the mechanism of resistance in detail and to inform the design of next-generation protease inhibitors.

Effects of accelerated rehabilitation exercise after mbo on ankle function and strength in ankle instability / Efectos del ejercicio de rehabilitación acelerada después de tbm sobre la función y la fuerza en la inestabilidad del tobillo / Efeitos do exercício de reabilitação acelerada depois de tbm sobre a função e a força na instabilidade do tornozelo

ABSTRACT Background: Taekwondo athletes with lateral chronic ankle instability (LCAI) may experience difficulties with proprioception and ankle muscle strength. After surgery, a conventional rehabilitation exercise program can be ineffective, as it may not restore proprioception or muscle strength and, thus, result in minimal improvement. Objective: This study aimed to assess the effects of an ARE program following MBO on the proprioception, isokinetic strength, and endurance of ankle. Methods: Thirty athletes diagnosed with LCAI underwent MBO. They were divided into the ARE group (n=15) and the control group (CON, n=15). The ARE group performed early ARE exercises six times per week for 4 weeks. The parameters applied to test ankle muscle strength were angular speed of 30°/sec and 180°/sec (inversion: Inv. and eversion: Eve., respectively). Proprioception was measured as being able to sense a joint position of 15° of Inv. and 5° of Eve. Results: Significant differences were observed between the ARE and CON groups in ankle strength 30°/sec (Inv.: p<0.001, Eve.: p<0.001), 180°/sec (Inv.: p<0.001, Eve.: p<0.001), and proprioception at 15° (Inv.: p<0.001) and 5° (Eve.: p<0.001). Conclusions: We recorded significant short-term effects from early ARE in Taekwondo athletes with LCAI after MBO. The results provide reference data for the sports rehabilitator or ATC in evaluating the rehabilitation phase and informing patients about expectations after MBO in terms of performance status and the timing of return to sports. Level of evidence III; Therapeutic studies-Investigation of treatment outcomes - Case-control Study.

RESUMEN Introducción: Los atletas de Tae Kwon Do con inestabilidad lateral crónica del tobillo (ILCT) pueden presentar dificultades con la propiocepción y la fuerza muscular del tobillo. Después de la cirugía, un programa convencional de ejercicios de rehabilitación puede ser ineficaz porque la propiocepción o la fuerza muscular pueden no restablecerse y, por tanto, dar lugar a una mejora mínima. Objetivos: Este estudio tuvo como objetivo evaluar los efectos de un programa de ERA después de TBM sobre la propiocepción, la fuerza isocinética y la resistencia del tobillo. Métodos: Treinta atletas diagnosticados con ILCT fueron sometidos a TBM. Se dividieron en grupo ERA (n=15) y grupo de control CON (n = 15). El grupo de ERA realizó ejercicios ERA precoces seis veces por semana durante 4 semanas. Los parámetros aplicados para probar la fuerza muscular del tobillo fueron la velocidad angular de 30°/s y 180°/s (inversión: Inv y eversión: Eve, respectivamente). La propiocepción se midió por la capacidad de sentir la posición articular de 15° de Inv y 5° de Eve. Resultados: Se observaron diferencias significativas entre los grupos ERA y CON en la fuerza del tobillo a 30°/s (Inv: p < 0,001, Eve: p < 0,001), 180°/s (Inv: p < 0,001, Eve: p < 0,001), y la propiocepción a 15° (Inv: p < 0,001) y 5° (Eve: p < 0,001). Conclusiones: Registramos efectos significativos a corto plazo con ERA precoz en atletas de Tae Kwon Do con ILCT después de TBM. Los resultados proporcionan datos de referencia para el rehabilitador deportivo o ATC (Certified athletic trainer) a la hora de evaluar la fase de rehabilitación e informar a los pacientes sobre las expectativas después de la TBM en cuanto a las condiciones de desempeño y el momento de retorno al deporte. Nivel de evidencia III; Estudios terapéuticos - Investigación de los resultados del tratamiento - Estudio de caso-control

RESUMO Introdução: Os atletas de taekwondo com instabilidade lateral crônica do tornozelo (ILCT) podem apresentar dificuldades com propriocepção e força muscular do tornozelo. Depois da cirurgia, um programa de exercícios de reabilitação convencional pode ser ineficaz, porque a propriocepção ou a força muscular podem não se restaurar e, portanto, resultar em melhora mínima. Objetivos: Este estudo teve como objetivo avaliar os efeitos de um programa de ERA depois de TBM sobre a propriocepção, força isocinética e resistência do tornozelo. Métodos: Trinta atletas com diagnóstico de ILCT foram submetidos à TBM. Eles foram divididos em grupo ERA (n=15) e grupo controle CON (n = 15). O grupo ERA realizou exercícios ERA precoces seis vezes por semana durante 4 semanas. Os parâmetros aplicados para testar a força muscular do tornozelo foram velocidade angular de 30°/s e 180°/s (inversão: Inv e eversão: Eve, respectivamente). A propriocepção foi medida pela capacidade de sentir a posição articular de 15° de Inv e 5° de Eve Resultados: Foram observadas diferenças significativas entre os grupos ERA e CON na força do tornozelo 30°/s (Inv: p < 0,001, Eve: p < 0,001), 180°/s (Inv: p < 0,001, Eve: p < 0,001), e propriocepção a 15° (Inv: p < 0,001) e 5° (Eve: p < 0,001). Conclusões: Registramos efeitos de curto prazo significativos com ERA precoce em atletas de taekwondo com ILCT depois da TBM. Os resultados fornecem dados de referência para o reabilitador esportivo ou ATC (Certified athletic trainer) na avaliação da fase de reabilitação e informar os pacientes sobre as expectativas depois da TBM em termos de condições de desempenho e momento de retorno ao esporte. Nível de evidência III; Estudos terapêuticos - Investigação dos resultados do tratamento - Estudo de caso-controle.