Assessment of the reversibility of resistance in the absence of antibiotics and its relationship with the resistance gene's fitness cost: a genetic study with mcr-1.

BACKGROUND: The intensive use of antibiotics has resulted in strong natural selection for the evolution of antimicrobial resistance (AMR), but whether, and under what circumstances, the removal of antibiotics would result in a rapid reduction in AMR has been insufficiently explored. We aimed to test the hypothesis that in the simple, yet common, case of AMR conferred by a single gene, removing antibiotics would quickly reduce the prevalence of resistance if the AMR gene imposes a high fitness cost and costless resistance is extremely rare among its proximal mutants. METHODS: In this genetic study, to test our hypothesis, we used the mcr-1 gene in Escherichia coli, which confers resistance to the last-resort antibiotic colistin, as a model. A high-throughput reverse genetics approach was used to evaluate mcr-1 variants for their fitness cost and resistance levels relative to a non-functional construct, by measuring relative growth rates in colistin-free media and at 2 µg/mL and 4 µg/mL colistin. We identified costless resistant mcr-1 mutants, and examined their properties within the context of the sequential organisation of mcr-1's functional domains as well as the evolutionary accessibility of these mutations. Finally, a simple population genetic model incorporating the measured fitness cost was constructed and tested against previously published real-world data of mcr-1 prevalence in colonised inpatients in China since the 2017 colistin ban in fodder additives. FINDINGS: We estimated the relative growth rates of 14 742 mcr-1 E coli variants (including the wild type), 3449 of which were single-nucleotide mutants. E coli showed 73·8% less growth per 24 h when carrying wild-type mcr-1 compared with the non-functional construct. 6252 (42·4%) of 14 741 mcr-1 mutants showed colistin resistance accompanied by significant fitness costs, when grown under 4 µg/mL colistin selection. 43 (0·3%) mcr-1 mutants exhibited costless resistance, most of which contained multiple mutations. Among the 3449 single mutants of mcr-1, 3433 (99·5%) had a fitness cost when grown in colistin-free media, with a mean relative growth of 0·305 (SD 0·193) compared with the non-functional variant. 3059 (88·7%) and 1833 (53·1%) of 3449 single mutants outgrew the non-functional mcr-1 in the presence of 2 µg/mL and 4 µg/mL colistin, respectively. Single mutations that gave rise to costless mutants were rare in all three domains of mcr-1 (transmembrane domain, flexible linker, and catalytic domain), but the linker domain was enriched with cost-reducing and resistance-enhancing mutations and depleted with cost-increasing mutations. The population genetics model based on the experimental data accurately predicts the rapid decline in mcr-1 prevalence in real-world data. INTERPRETATION: Many identified costless resistant variants that consist of multiple mutations are unlikely to evolve easily in nature. These findings for colistin and mcr-1 might be applicable to other cases in which AMR entails a substantial fitness cost that cannot be mitigated in proximal mutants. FUNDING: National Natural Science Foundation of China, and National Key Research and Development Program of China.

Land degradation neutrality assessment and factors influencing it in China's arid and semiarid regions.

The ecosystems in China's arid and semiarid regions are notably fragile and experiencing dramatic land degradation. At the 12th Conference of the Parties (COP12) to the United Nations Convention to Combat Desertification (UNCCD) in October 2015, a definition for land degradation neutrality (LDN) was proposed and subsequently integrated into the Sustainable Development Goals (SDGs). Research on LDN has developed in terms of conceptual framework constructions, quantitative assessments, and empirical studies. However, LDN and its drivers must be clarified in China's arid and semiarid regions since some representative processes have yet to be fully considered in the assessment. Here, we develop an LDN indicator system specialised for the area, assess their LDN status, and determine the impacts of human activities and climate change on LDN. Our research aims to refine the LDN indicator system tailored for China's arid and semiarid regions by incorporating the trends of wind and water erosion. We also identify the influence of human activity and climate change on LDN, which provides insightful strategies for ecological restoration and sustainable development in drylands with climate-sensitive ecosystems. The results show that: (1) In 2020, more than half of areas of China's arid and semiarid regions achieved LDN, with more pronounced success in the southeastern areas compared to the central regions. (2) For LDN drivers, elevation shows negligible influence on LDN, whereas increased temperature promotes LDN achievement. Conversely, factors like vapour pressure deficit and v-direction wind speed hinder it. In conclusion, China's arid and semiarid regions achieved LDN, and the dominant factor that substantially influences LDN varies across geographical zones, with higher wind speeds and elevated GDP levels generally obstructing LDN in most areas.

High-throughput mutagenesis and screening approach for the identification of drug-resistant mutations in the rifampicin resistance-determining region of mycobacteria.

Rifampicin is the most powerful first-line antibiotic for tuberculosis, which is caused by Mycobacterium tuberculosis. Although accumulating evidence from sequencing data of clinical M. tuberculosis isolates suggested that mutations in the rifampicin-resistance-determining region (RRDR) are strongly associated with rifampicin resistance, the comprehensive characterisation of RRDR polymorphisms that confer this resistance remains challenging. By incorporating I-SceI sites for I-SceI-based integrant removal and utilizing an L5 swap strategy, we efficiently replaced the integrated plasmid with alternative alleles, making mass allelic exchange feasible in mycobacteria. Using this method to establish a fitness-related gain-of function screen, we generated a mutant library that included all single-amino-acid mutations in the RRDR, and identified the important positions corresponding to some well-known rifampicin-resistance mutations (Q513, D516, S522, H525, R529, S531). We also detected a novel two-point mutation located in the RRDR confers a fitness advantage to M. smegmatis in the presence or absence of rifampicin. Our method provides a comprehensive insight into the growth phenotypes of RRDR mutants and should facilitate the development of anti-tuberculosis drugs.

A latent class analysis of depressive symptoms among rural Chinese adolescents and their association with psychological resilience.

Numerous studies have demonstrated the heterogeneity of depressive symptoms, but few studies have focused on the heterogeneity of depressive symptoms among rural Chinese adolescents. In November to December 2022, multistage sampling was employed to administer questionnaires to 1,816 rural adolescents aged 11-19 years from six schools in Henan Province, China. Depressive symptoms were measured using the Chinese version of the Children's Depression Inventory Scale. Latent class analysis (LCA) was utilized to identify subgroups of depressive symptoms. The investigation of subgroup characteristics and associated factors was conducted through χ2 tests, ANOVA, and multinomial logistic regression analyses. The findings revealed a 24.24 % detection rate of depressive symptoms among Chinese rural adolescents. LCA analysis of responses to the 27 items in the Depressive Symptoms Scale led to the classification of depressive symptoms into four subgroups based on severity: "no depressive symptoms group" (22.5 %), a "low depressive symptoms group" (35.7 %), a "transition group" (31.6 %), and a "high depressive symptoms group" (10.2 %). Gender, grade level, academic performance, academic stress, family environment, and level of psychological resilience are associated factors for subgroups of depressive symptoms among rural adolescents. There should be increased training of rural educators to enable early recognition of depressive characteristics and risk factors, facilitating targeted prevention and intervention strategies.

The Impact of Different Intensities of Physical Activity on Serum Urate and Gout: A Mendelian Randomization Study.

Physical activity is a potential protective factor against gout, but the role of exercise intensity in this context remains unclear. To overcome the limitations of observational studies in causal inference, this study employed a two-sample Mendelian randomization approach to explore the impact of different genetically proxied/predicted intensities of physical activity on serum urate concentration and the incidence of gout. Our data related to physical activity, serum urate, and gout were obtained from the UK Biobank, the Global Urate Genetics Consortium (GUGC), and the FinnGen dataset, respectively. Walking was included as representative of typical low-intensity physical activity in the analysis, and the other two types were moderate and vigorous physical activities. The estimation methods we used included the inverse-variance-weighted (IVW) method, MR-Egger regression, weighted-median method, simple-mode method, and weighted-mode method. Sensitivity analyses involved Rucker's framework, Cochran's Q test, funnel plots, MR-PRESSO outlier correction, and leave-one-out analysis. We found suggestive evidence from the inverse-variance-weighted method that moderate physical activity was a potential factor in reducing the incidence of gout (OR = 0.628, p = 0.034), and this association became more substantial in our subsequent sensitivity analysis (OR = 0.555, p = 0.006). However, we observed no distinctive effects of physical activity on serum urate concentration. In conclusion, our study supports some findings from observational studies and emphasizes the preventive role of moderate physical activity against gout. Given the limitations of the existing datasets, we call for future reexamination and expansion of our findings using new GWAS data.

A new variant of the colistin resistance gene MCR-1 with co-resistance to ß-lactam antibiotics reveals a potential novel antimicrobial peptide.

The emerging and global spread of a novel plasmid-mediated colistin resistance gene, mcr-1, threatens human health. Expression of the MCR-1 protein affects bacterial fitness and this cost correlates with lipid A perturbation. However, the exact molecular mechanism remains unclear. Here, we identified the MCR-1 M6 variant carrying two-point mutations that conferred co-resistance to ß-lactam antibiotics. Compared to wild-type (WT) MCR-1, this variant caused severe disturbance in lipid A, resulting in up-regulation of L, D-transpeptidases (LDTs) pathway, which explains co-resistance to ß-lactams. Moreover, we show that a lipid A loading pocket is localized at the linker domain of MCR-1 where these 2 mutations are located. This pocket governs colistin resistance and bacterial membrane permeability, and the mutated pocket in M6 enhances the binding affinity towards lipid A. Based on this new information, we also designed synthetic peptides derived from M6 that exhibit broad-spectrum antimicrobial activity, exposing a potential vulnerability that could be exploited for future antimicrobial drug design.

Improving the High-Temperature Energy Storage Performance of Epoxy Films: Moderately Reducing Unsaturation for Extremely High Efficiency.

The rapid development of renewable energy systems, electric vehicles, and pulsed equipment requires energy storage media to have a high energy storage density and efficiency in a wide temperature range. The state-of-the-art biaxially oriented polypropylene (BOPP) film is insufficient to meet the growing demand for energy storage devices due to its low energy storage density and working temperature, which make it a research hotspot for developing dielectric energy storage materials. In this manuscript, based on the epoxy materials that have been shown as a potential energy storage medium, we aim to reduce the influence of the benzene ring delocalization structure on the energy storage losses and enhance the efficiency by gradually replacing them with cyclohexane structures to adjust the segment unsaturation of epoxy materials. The results show that by partially reducing the unsaturation of the curing agent, the epoxy material achieves an excellent high-temperature energy storage density of 2.21 J/cm3 at 150 °C and 300 MV/m while maintaining an extremely high energy storage efficiency of 99.2%. Leakage current density and high-voltage dielectric spectroscopy tests confirm that a moderate reduction of the segment unsaturation of epoxy materials can greatly inhibit polarization loss at high temperatures, which may explain their high energy storage efficiency.

Genomic Diversity Illuminates the Environmental Adaptation of Drosophila suzukii.

Biological invasions carry substantial practical and scientific importance, and represent natural evolutionary experiments on contemporary timescales. Here, we investigated genomic diversity and environmental adaptation of the crop pest Drosophila suzukii using whole-genome sequencing data and environmental metadata for 29 population samples from its native and invasive range. Through a multifaceted analysis of this population genomic data, we increase our understanding of the D. suzukii genome, its diversity and its evolution, and we identify an appropriate genotype-environment association pipeline for our data set. Using this approach, we detect genetic signals of local adaptation associated with nine distinct environmental factors related to altitude, wind speed, precipitation, temperature, and human land use. We uncover unique functional signatures for each environmental variable, such as a prevalence of cuticular genes associated with annual precipitation. We also infer biological commonalities in the adaptation to diverse selective pressures, particularly in terms of the apparent contribution of nervous system evolution to enriched processes (ranging from neuron development to circadian behavior) and to top genes associated with all nine environmental variables. Our findings therefore depict a finer-scale adaptive landscape underlying the rapid invasion success of this agronomically important species.

The Effect of Various Training Variables on Developing Muscle Strength in Velocity-based Training: A Systematic Review and Meta-analysis.

Velocity-based training is an advanced auto-regulation method that uses objective indices to dynamically regulate training loads. However, it is unclear currently how to maximize muscle strength with appropriate velocity-based training settings. To fill this gap, we conducted a series of dose-response and subgroup meta-analyses to check the effects of training variables/parameters, such as intensity, velocity loss, set, inter-set rest intervals, frequency, period, and program, on muscle strength in velocity-based training. A systematic literature search was performed to identify studies via PubMed, Web of Science, Embase, EBSCO, and Cochrane. One repetition maximum was selected as the outcome to indicate muscle strength. Eventually, twenty-seven studies with 693 trained individuals were included in the analysis. We found that the velocity loss of 15 to 30%, the intensity of 70 to 80%1RM, the set of 3 to 5 per session, the inter-set rest interval of 2 to 4 min, and the period of 7 to 12 weeks could be appropriate settings for developing muscle strength. Three periodical programming models in velocity-based training, including linear programming, undulating programming, and constant programming, were effective for developing muscle strength. Besides, changing periodical programming models around every 9 weeks may help to avoid a training plateau in strength adaption.

Bacillus licheniformis prevents and reduces anxiety-like and depression-like behaviours.

As common mental disorders, depression and anxiety impact people all around the world. Recent studies have found that the gut microbiome plays an important role in mental health. It is becoming possible to treat mental disorders by regulating the composition of the gut microbiota. Bacillus licheniformis is a probiotic used to treat gut diseases through balancing the gut microbiome during lasting years. Considering the role of gut microbiota in the gut-brain axis, this study used chronic unpredictable mild stress (CUMS) model rats to explore whether Bacillus licheniformis can prevent and treat depression and anxiety. We found that B. licheniformis reduced the depressive-like and anxiety-like behaviours of the rats during the CUMS process. Meanwhile, B. licheniformis changed the gut microbiota composition; increased the short chain fatty acids (SCFAs) in the colon, decreased kynurenine, norepinephrine, and glutamate levels; and increased the tryptophan, dopamine, epinephrine, and γ-aminobutyric acid (GABA) in the brain. After correlation analysis, we found Parabacteroides, Anaerostipes, Ruminococcus-2, and Blautia showed significant correlation with neurotransmitters and SCFAs, indicating the gut microbiome plays an important role in B. licheniformis reducing depressive-like behaviours. Therefore, this study suggested B. licheniformis may prevent depressive-like and anxiety-like behaviours while regulating the gut microbiota composition and increasing the SCFA levels in the colon to alter the levels of the neurotransmitters in the brain. KEY POINTS: • B. licheniformis reduced depressive-like and anxiety-like behaviours induced by the chronic unpredictable mild stress. • GABA levels in the brain are assonated with B. licheniformis regulating depressive-like and anxiety-like behaviours. • Gut microbiota composition alteration followed by metabolic changes may play a role in the GABA levels increase.

Positive mood-related gut microbiota in a long-term closed environment: a multiomics study based on the "Lunar Palace 365" experiment.

BACKGROUND: Psychological health risk is one of the most severe and complex risks in manned deep-space exploration and long-term closed environments. Recently, with the in-depth research of the microbiota-gut-brain axis, gut microbiota has been considered a new approach to maintain and improve psychological health. However, the correlation between gut microbiota and psychological changes inside long-term closed environments is still poorly understood. Herein, we used the "Lunar Palace 365" mission, a 1-year-long isolation study in the Lunar Palace 1 (a closed manned Bioregenerative Life Support System facility with excellent performance), to investigate the correlation between gut microbiota and psychological changes, in order to find some new potential psychobiotics to maintain and improve the psychological health of crew members. RESULTS: We report some altered gut microbiota that were associated with psychological changes in the long-term closed environment. Four potential psychobiotics (Bacteroides uniformis, Roseburia inulinivorans, Eubacterium rectale, and Faecalibacterium prausnitzii) were identified. On the basis of metagenomic, metaproteomic, and metabolomic analyses, the four potential psychobiotics improved mood mainly through three pathways related to nervous system functions: first, by fermenting dietary fibers, they may produce short-chain fatty acids, such as butyric and propionic acids; second, they may regulate amino acid metabolism pathways of aspartic acid, glutamic acid, tryptophan, etc. (e.g., converting glutamic acid to gamma-aminobutyric acid; converting tryptophan to serotonin, kynurenic acid, or tryptamine); and third, they may regulate other pathways, such as taurine and cortisol metabolism. Furthermore, the results of animal experiments confirmed the positive regulatory effect and mechanism of these potential psychobiotics on mood. CONCLUSIONS: These observations reveal that gut microbiota contributed to a robust effect on the maintenance and improvement of mental health in a long-term closed environment. Our findings represent a key step towards a better understanding the role of the gut microbiome in mammalian mental health during space flight and provide a basis for future efforts to develop microbiota-based countermeasures that mitigate risks to crew mental health during future long-term human space expeditions on the moon or Mars. This study also provides an essential reference for future applications of psychobiotics to neuropsychiatric treatments. Video Abstract.

The causal effect of physical activity intensity on COVID-19 susceptibility, hospitalization, and severity: Evidence from a mendelian randomization study.

The protection of physical activity (PA) against COVID-19 is a rising research interest. However, the role of physical activity intensity on this topic is yet unclear. To bridge the gap, we performed a Mendelian randomization (MR) study to verify the causal influence of light and moderate-to-vigorous PA on COVID-19 susceptibility, hospitalization, and severity. The Genome-Wide Association Study (GWAS) dataset of PA (n = 88,411) was obtained from the UK biobank and the datasets of COVID-19 susceptibility (n = 1,683,768), hospitalization (n = 1,887,658), and severity (n = 1,161,073) were extracted from the COVID-19 Host Genetics Initiative. A random-effect inverse variance weighted (IVW) model was carried out to estimate the potential causal effects. A Bonferroni correction was used for counteracting. The problem of multiple comparisons. MR-Egger test, MR-PRESSO test, Cochran's Q statistic, and Leave-One-Out (LOO) were used as sensitive analysis tools. Eventually, we found that light PA significantly reduced the risk of COVID-19 infection (OR = 0.644, 95% CI: 0.480-0.864, p = 0.003). Suggestive evidence indicated that light PA reduced the risks of COVID-19 hospitalization (OR = 0.446, 95% CI: 0.227 to 0.879, p = 0.020) and severe complications (OR = 0.406, 95% CI: 0.167-0.446, p = 0.046). By comparison, the effects of moderate-to-vigorous PA on the three COVID-19 outcomes were all non-significant. Generally, our findings may offer evidence for prescribing personalized prevention and treatment programs. Limited by the available datasets and the quality of evidence, further research is warranted to re-examine the effects of light PA on COVID-19 when new GWAS datasets emerge.

The Effect of Velocity Loss on Strength Development and Related Training Efficiency: A Dose-Response Meta-Analysis.

The velocity loss method is often used in velocity-based training (VBT) to dynamically regulate training loads. However, the effects of velocity loss on maximum strength development and training efficiency are still unclear. Therefore, we conducted a dose-response meta-analysis aiming to fill this research gap. A systematic literature search was performed to identify studies on VBT with the velocity loss method via PubMed, Web of Science, Embase, EBSCO, and Cochrane. Controlled trials that compared the effects of different velocity losses on maximum strength were considered. One-repetition maximum (1RM) gain and 1RM gain per repetition were the selected outcomes to indicate the maximum strength development and its training efficiency. Eventually, nine studies with a total of 336 trained males (training experience/history ≥ 1 year) were included for analysis. We found a non-linear dose-response relationship (reverse U-shaped) between velocity loss and 1RM gain (pdose-response relationship < 0.05, pnon-linear relationship < 0.05). Additionally, a negative linear dose-response relationship was observed between velocity loss and 1RM gain per repetition (pdose-response relationship < 0.05, pnon-linear relationship = 0.23). Based on our findings, a velocity loss between 20 and 30% may be beneficial for maximum strength development, and a lower velocity loss may be more efficient for developing and maintaining maximum strength. Future research is warranted to focus on female athletes and the interaction of other parameters.

Can Intelligence Affect Alcohol-, Smoking-, and Physical Activity-Related Behaviors? A Mendelian Randomization Study.

People with high levels of intelligence are more aware of risk factors, therefore choosing a healthier lifestyle. This assumption seems reasonable, but is it true? Previous studies appear to agree and disagree. To cope with the uncertainty, we designed a mendelian randomization (MR) study to examine the causal effects of genetically proxied intelligence on alcohol-, smoking-, and physical activity (PA)-related behaviors. We obtained genome-wide association study (GWAS) datasets concerning these variables from separate studies or biobanks and used inverse-variance weighted (IVW) or MR-Egger estimator to evaluate the causal effects according to an MR protocol. The MR-Egger intercept test, MR-PRESSO, and funnel plots were employed for horizontal pleiotropy diagnosis. The Steiger test (with reliability test), Cochran's Q test, MR-PRESSO, and leave-one-out method were employed for sensitivity analysis. We found significant or potential effects of intelligence on alcohol dependence (OR = 0.749, p = 0.003), mental and behavioral disorders due to alcohol (OR = 0.814, p = 0.009), smoking (OR = 0.585, p = 0.005), and smoking cessation (OR = 1.334, p = 0.001). Meanwhile, we found significant or potential effects on walking duration (B = -0.066, p < 0.001), walking frequency (B = -0.055, p = 0.031), moderate PA frequency (B = -0.131, p < 0.001), and vigorous PA frequency (B = -0.070, p = 0.001), but all in a negative direction. In conclusion, our findings reinforce some existing knowledge, indicate the complexity of the health impacts of human intelligence, and underline the value of smoking and alcohol prevention in less intelligent populations. Given the existing limitations in this study, particularly the potential reverse causality in some estimations, re-examinations are warranted in future research.

Influences of water level fluctuation on water exchange and nutrient distribution in a bay: Evidence from the Xiangxi Bay, Three Gorges Reservoir.

Following the Three Gorges Reservoir (TGR) impoundment, many tributaries were turned into bays; hydrodynamic conditions of TGR profoundly changed the residence time, temperature, and nutrient distributions of bays, and nutrient enrichment occurred in these bays. However, little research has been done on the effects of water level qqfluctuations (WLFs) of TGR on the bay. In this study, Xiangxi Bay (XXB), one of the tributaries of TGR, was selected as the delegate to construct and calibrate a two-dimensional hydrodynamic-temperature-tracer-water quality model based on the CE-QUAL-W2. The results were the following: 1) In spring, as total nitrogen (TN) in the TGR tended to be higher than that in the XXB, the downward WLF increased water exchange, TGR-XXB nutrient flux and TN in the epilimnion of the XXB, and decreased the water exchange and TN in the hypolimnion of the XXB. The upward WLF did the opposite. The situation would be reversed in autumn. 2) Under a larger magnitude or a shorter period of WLF, its corresponding effects on the water exchange and TN increased. 2) Both the downward and upward modes of WLF helped to decrease the thermal stratification of XXB. 4) The upward/downward WLF could be used to decrease the epilimnetic TN of XXB in spring/autumn, and was suggested to reduce the local algal bloom. The WLFs by the TGR regulation could profoundly change the water exchange and nutrient distribution in the bay, which helped to control nutrient concentrations and prevent algal blooms.

Using structural equation modeling to explore the influences of physical activity, mental health, well-being, and loneliness on Douyin usage at bedtime.

Douyin is the Chinese version of TikTok. Using Douyin at bedtime is a very common behavior among Douyin users. However, the reasons why users like using Douyin before sleep are yet unclear. We conducted a cross-section survey from January 1st to January 16th, 2023 to capture data to examine the associations of depression, anxiety, life satisfaction, well-being, loneliness, and physical activity with Douyin usage at bedtime. The mediation role of insomnia in these associations was examined. A total of 3,392 participants who met the inclusion criteria were included for analysis. Our structural equation modeling analysis showed that depression on (ß = 0.08; p < 0.05), anxiety (ß = 0.06; p < 0.05), and loneliness (ß = 0.14; p < 0.05) were directly associated with increased Douyin usage at bedtime, and were also indirectly associated with Douyin usage through insomnia (depression: ß = 0.18; p < 0.05, anxiety: ß = 0.16; p < 0.05, and loneliness: ß = 0.12; p < 0.05). Life satisfaction (ß = -0.05; p < 0.05) and well-being (ß = -0.20; p < 0.05) were directly associated with decreased Douyin usage at bedtime, and were also indirectly associated with Douyin usage through insomnia (life satisfaction: ß = -0.09; p < 0.05, and well-being: ß = -0.11; p < 0.05). However, physical activity was unexpectedly associated with increased Douyin usage at bedtime (ß = 0.20; p < 0.05). In conclusion, our findings shed new light on the specific reasons why Douyin users like using Douyin at bedtime.

Can Blood Flow Restriction Training Benefit Post-Activation Potentiation? A Systematic Review of Controlled Trials.

(1) Background: post-activation potentiation (PAP) plays an essential role in enhancing athletic performance. Various conditioning activities (CAs) have been developed to generate PAP before training or competitions. However, whether extra equipment can enhance the effectiveness of CAs is understudied. Hence, this systematic review aims to introduce and examine the effectiveness of blood flow restriction-based conditioning activities (BFR-CAs). (2) Methods: a literature search was conducted via Web of Science, PubMed, SPORTDiscus, and CNKI (a Chinese academic database). The systematic review included the literature concerning BFR-CAs and non-BFR-CAs. The methodological quality of included studies was considered to be "moderate quality" and "good quality" based on the Physiotherapy Evidence Database Scale. (3) Results: five studies were included in this study. Four studies were on lower limb strength training, and three of them suggested a greater PAP in BFR-CAs than in non-BFR counterparts. One study on upper limb strength training also supported the advantage of BFR-CAs. (4) Conclusions: BFR-CAs may be an emerging and promising strategy to generate PAP. Compared with non-BFR-CAs, BFR-CAs might be more efficient and practical for inexperienced sports people or athletes in non-power sports.

A CRISPR-guided mutagenic DNA polymerase strategy for the detection of antibiotic-resistant mutations in M. tuberculosis.

A sharp increase in multidrug-resistant tuberculosis (MDR-TB) threatens human health. Spontaneous mutation in essential gene confers an ability of Mycobacterium tuberculosis resistance to anti-TB drugs. However, conventional laboratory strategies for identification and prediction of the mutations in this slowly growing species remain challenging. Here, by combining XCas9 nickase and the error-prone DNA polymerase A from M. tuberculosis, we constructed a CRISPR-guided DNA polymerase system, CAMPER, for effective site-directed mutagenesis of drug-target genes in mycobacteria. CAMPER was able to generate mutagenesis of all nucleotides at user-defined loci, and its bidirectional mutagenesis at nick sites allowed editing windows with lengths up to 80 nucleotides. Mutagenesis of drug-targeted genes in Mycobacterium smegmatis and M. tuberculosis with this system significantly increased the fraction of the antibiotic-resistant bacterial population to a level approximately 60- to 120-fold higher than that in unedited cells. Moreover, this strategy could facilitate the discovery of the mutation conferring antibiotic resistance and enable a rapid verification of the growth phenotype-mutation genotype association. Our data demonstrate that CAMPER facilitates targeted mutagenesis of genomic loci and thus may be useful for broad functions such as resistance prediction and development of novel TB therapies.