The clinical relevance of echocardiographic measurements of right heart size and function in dogs with pulmonary hypertension (PH) is unknown.To determine if echocardiographic measurements of right heart size and right ventricular (RV) function are associated with survival times in dogs with PH.Eighty‐two client‐owned dogs.Retrospective study where data from medical records and baseline echocardiographic examinations were collected and measured in a standardized manner. Owners or primary veterinarians were contacted for outcome data.P = .008) more common in dogs with PH not secondary to left heart disease  compared to dogs with PH secondary to LHD (14%) but median survival times  were not significantly different (P = .78). Right atrial area , TAPSE < 3.23 mm/kg0.284 , and right heart failure  were independently associated with shorter survival time (P ≤ .04).Enlargement of the right atrium (88%), RV (69%), and pulmonary artery (72%) was common. One‐third of the cases had reduced RV function quantified by two‐dimensional echocardiography‐derived tricuspid annular plane systolic excursion (TAPSE). Decreased TAPSE was significantly (0.284), and right heart failure offer clinically relevant prognostic information in dogs with PH. Results support the quantitative assessment of right heart size and function in dogs with PH.Right atrial area, RV function (TAPSE < 3.23 mm/kg AT/ETacceleration time of pulmonary artery blood flow indexed to ejection timeLAleft atrium/atrialLA/Aoleft atrium‐to‐aortic root dimensionLADleft atrial dimensionLHDleft heart diseaseLVIDdleft ventricular internal dimension at end‐diastoleMMVDmyxomatous mitral valve diseasePGpressure gradientPHpulmonary hypertensionPV/Aopulmonary valve indexed to the aortic rootRAright atrium/atrialRAAright atrial areaRPADright pulmonary artery distensibilityRVright ventricle/ventricularRVAdright ventricular area at end‐diastoleRVIDdright ventricular internal dimension at end‐diastoleTAPSEtricuspid annular plane systolic excursionTRPGtricuspid regurgitation pressure gradientTRVtricuspid regurgitation velocity1Pulmonary hypertension (PH) is a secondary complication of many commonly encountered cardiopulmonary disorders and is an increasingly recognized clinically important finding in dogs.2) is the primary metric used to estimate pulmonary arterial pressure and helps to assess the echocardiographic probability a dog has PH.Veterinarians are heavily reliant on echocardiography to provide a clinical diagnosis of PH. Doppler echocardiography‐derived tricuspid regurgitation velocity (TRV) and thus tricuspid regurgitation PG calculated using the simplified Bernoulli equation , if they had intracardiac heartworms and underwent interventional heartworm retrieval, if they were diagnosed with congenital cardiac shunt, or if they had a sustained or clinically important arrhythmia .2.2Dogs were classified with PH secondary to LHD if they had LHD  and unequivocal echocardiographic left atrial enlargement2.32. Regurgitant jets were measured at the dense outer edge of the velocity profile and measurements of fine linear signals (feathered edge) were avoided.All echocardiographic examinations were performed by one of the authors (a board‐certified cardiologist) or a cardiology resident trained by the author. All echocardiographic assessments, measurements, and calculations were performed by a single investigator (L. C. V.) at an off‐cart workstation . All echocardiographic studies were measured in the same order. The TRV recorded for study purposes was measured last. It was measured using continuous‐wave Doppler recordings from the imaging plane that permitted the fastest jet profile. The TRV was converted to a tricuspid regurgitation pressure gradient (TRPG) using the simplified Bernoulli equation: TRPG = 4 Linear measurements of the left atrium (LA), left ventricle, and RV were performed from the right parasternal 4‐chamber long‐axis imaging planes. Maximum LAD was measured midchamber at end (ventricular)‐systole with a line extending from the region of the fossa ovale to the internal reflection of the hyperechoic pericardium in the far field approximately parallel to the mitral annulus.Left atrial and aortic dimensions were measured from the right parasternal short‐axis view at the level of the aortic root.The right parasternal short‐axis basilar view optimized for the RV outflow tract that permitted adequate visualization of the pulmonary trunk and right pulmonary artery was used for the following measurements. Diameter of the pulmonary valve was measured in early diastole and indexed to the aorta root measurement (PV/Ao) as described above for LA/Ao. Pulmonary regurgitation velocity was measured in early diastole, if present. The ratio of acceleration time to ejection time (AT/ET) of pulsed‐wave Doppler recordings of blood flow was measured at the level of the pulmonary valve.A left apical 4‐chamber view, usually optimized for the right heart, was utilized for measurements of right atrial and ventricular size and RV function. Maximum right atrial (end‐systolic) and ventricular (end‐diastolic) area  were quantified using planimetry as previously described.2.4Study investigators contacted dog owners, primary care veterinarians, or both to determine the outcome of each dog  if this could not be gathered from our hospital's medical records. Survival time was counted from the day of the echocardiographic diagnosis of PH to either the day of death or, if alive, to the start of data collection . Because of the challenge of attributing death directly to PH, the end point of the study was death/euthanasia of any cause .2.5t test. If normality testing failed, continuous data were reported as median (interquartile range) and compared with a Mann‐Whitney test. Categorical data were summarized as proportions and percentages and were compared with a Chi‐squared test.Statistical analyses were performed using commercially available software . Descriptive statistics were generated and the Shapiro‐Wilk test was used for normality testing of continuous data. If normally distributed, continuous data were reported as mean (SD) and compared with an independent Effects of the following baseline clinical and echocardiographic variables on survival were evaluated using Kaplan‐Meier survival curves and log‐rank tests: (1) age, (2) PH secondary to LHD versus other causes, (3) right heart failure, (4) syncope, (5) TRV, (6) RAA, (7), RVAd, (8) TAPSE, (9) PV/Ao, (10) RPAD index, and (11) AT/ET. To evaluate for collinearity of the echocardiographic variables, all were compared using Spearman rank correlation analysis. Dogs that were lost to follow up were right‐censored after their last known contact. Dogs that were alive at the end of the study period were also right‐censored. Dogs that died before hospital discharge at the time of their echocardiographic diagnosis of PH were excluded from the survival analysis.2/kg0.71,2/kg0.62,0.284,P < .20. Two separate multivariable Cox proportional‐hazards regression analyses were performed; one using continuous echocardiographic predictor variables and one using the aforementioned clinical cutoffs . Multivariable analyses were performed using a forward stepwise approach. Following the multivariable Cox proportional‐hazards regression analysis on the continuous echocardiographic predictor variables, a post hoc analysis of RAA was performed in order to explore clinically relevant cutoffs for RAA. Cutoffs were explored by dividing RAA into groups based on quartiles, and the effect on survival of these cutoffs was evaluated using Kaplan‐Meier survival analysis and Log‐rank tests. For all analyses, statistical significance was set at P < .05.Cutoffs for continuous variables were chosen based on clinical relevance and the cited veterinary literature as follows: TRPG > 75 mm Hg (severe PH),33.1Eighty‐eight dogs met the inclusion criteria. However, 3 dogs were excluded because they had intracardiac heartworms and underwent interventional retrieval. Three dogs were excluded because of a congenital cardiac shunt; 1 with a left‐to‐right shunting atrial septal defect and 2 with a right‐to‐left shunting patent ductus arteriosus. Thus, 82 dogs were enrolled in our study and a summary of clinical findings is presented in Table Dirofilaria immitis antigen, of which, 1 tested positive, however no heartworms were visible with echocardiography. Left heart disease was documented in 28 of 82 dogs (34%) all of which were diagnosed with MMVD. The majority of dogs  had PH that was not associated with LHD.Forty dogs (49%) were tested for 0.284. Dogs with PH not secondary to LHD had significantly (P < .001) lower TAPSE (3.30 [2.74‐4.48] mm/kg0.284) compared to dogs with PH secondary to LHD (5.38 [4.11‐6.30] mm/kg0.284). Also, the proportion of dogs with reduced TAPSE (<3.23 mm/kg0.284) was significantly (P = .008) higher in dogs with PH not secondary to LHD  compared to dogs with PH secondary to LHD . Decreased RPAD and AT/ET were also common, occurring in 87% and 62% of dogs, respectively. Tricuspid annular plane systolic excursion, PV/Ao, and AT/ET were all significantly  when compared to dogs with PH not secondary to LHD . Conversely, RVIDd/LVIDd was significantly (P < .001) decreased in dogs with PH secondary to LHD (0.56 [0.36‐0.71]) when compared to dogs with PH not secondary to LHD (0.93 [0.70‐1.23]).A summary of the echocardiographic data for all dogs at the time of the diagnosis of PH is presented in Table 3.2P = .78) between median survival time for dogs with PH not secondary to LHD  compared to dogs with PH secondary to LHD .Four dogs were excluded from the survival analysis because of euthanasia before hospital discharge; all had PH not secondary to LHD. Sixty‐seven (86%) dogs died during the study period, which lasted 4 years and 8 months. Forty‐four dogs (66%) were euthanized and 23 (34%) died naturally. Eleven dogs were right‐censored; 5 were still alive and 6 were lost to follow up. Overall median survival time (n = 78 dogs) was 192 days . There was no significant difference . Of the 11 dichotomous variables evaluated as predictors of survival in the univariate analysis, right heart failure (P = .01), RVAd > 1.33 cm2/kg0.62 (P = .03), TAPSE < 3.23 mm/kg0.284 (P = .003), and RPAD index < 0.30 (P = .03) were significantly associated with worse outcomes  were carried forward into the multivariable analysis, only right heart failure (P = .02) and TAPSE < 3.23 mm/kg0.284 (P = .04) had independent adverse associations with survival time (Table P = .05), RAA (P = .007), RVAd (P = .02), RPAD index (P = .03), and AT/ET (P = .02) were significantly associated with worse outcomes (Table P = .02) was independently associated with survival time (Table 2/kg0.71), 50th (1.2 cm2/kg0.71), and 75th (1.5 cm2/kg0.71) percentiles on survival. Results showed that the cutoffs of RAA > 1.2 cm2/kg0.71  and RAA > 1.5 cm2/kg0.71  were significantly associated with survival  in addition to right heart failure were independently associated with reduced survival time in dogs with PH. These findings provide clinically relevant prognostic information and highlight the importance of quantifying right heart size and function  in dogs with PH related to any underlying cause.Our study showed that echocardiographic evidence of right heart enlargement is common in dogs with PH, regardless of the underlying etiology. Enlargement of the RA, RV, or PA was identified in at least 69% of all cases and RV systolic dysfunction was identified in one‐third of cases. Right ventricular function was significantly worse in dogs with PH not secondary to LHD when compared to dogs with PH secondary to LHD. However, survival times were not significantly different between dogs with PH secondary to LHD  and dogs with PH not secondary to LHD. Last, right atrial size (RAA) and decreased RV function (TAPSE < 3.23 mm/kgNumerous studies have documented that RA enlargement (increased RAA),0.284) was significantly less common in dogs with PH secondary to LHD/MMVD compared to PH due to other causes (PH not secondary to LHD). This represents an important finding that agrees with previous work evaluating TAPSE in dogs with PH and MMVD.Our study found that RV systolic dysfunction  are 2 times more likely to die than dogs with RAA 2/kg0.71; whereas dogs with RAA > 1.5 cm2/kg0.71 (75th percentile) are 3.6 times more likely to die than dogs with RAA 2/kg0.71. These cutoffs might be useful to help categorize RA enlargement  in a clinically relevant manner based on prognosis and relative risk of death in dogs with PH.We used 2 different multivariable Cox proportional‐hazards regression models, 1 using previously established clinically relevant dichotomous echocardiographic predictor variables and 1 using continuous echocardiographic predictor variables. The latter suggested that RAA as a continuous predictor variable is significantly associated with risk of death and demonstrates that risk of death increases (at a constant increment) as RA size increases (at a constant increment). To attempt to categorize RA size in a more clinically relevant manner, we performed a post hoc analysis on RAA based on categorization into quartiles. It showed that dogs with RAA > 1.2 cmOur results are not applicable to all dogs with PH and should be interpreted within the context of the population studied. Specifically, dogs with a congenital shunting lesion and dogs with intracardiac heartworms were excluded. Dogs with a congenital shunting lesion were excluded because their younger age might have skewed survival data. Also, some shunting lesions can be corrected or cured, further skewing survival data. Dogs with intracardiac heartworms were also excluded because of the challenges of assessing or estimating increased PA pressure . Also, in our experience, outcome of these cases can range from being very poor (death during heartworm extraction) to excellent (with successful extractions). Thus, our study population largely consisted of dogs with acquired and likely chronic and irreversible PH where it is unlikely the underlying disease can be cured or corrected.Given our study's retrospective design, all dogs did not have a consistent and thorough diagnostic evaluation nor could a definitive underlying cause of PH be determined to permit grouping of cases according to the cause of PH. Some causes of PH require invasive or impractical diagnostics for definitive diagnosis . Furthermore, classifying dogs into groups of PH is, in theory, intended to group dogs according to similar themes such as pathophysiology, treatment, and potentially prognosis.Results of our study should be interpreted within the context of its limitations. This was a retrospective study and therefore it lacked standardization of, for example, diagnostic evaluations and medications permitted. However, it should be noted that a minority of dogs enrolled in our study were taking medications known to affect the cardiovascular system (at most 32% were taking furosemide and only 4% were taking sildenafil). However, we recognize that this could have influenced our results to some degree. More importantly, as a retrospective study, treatment protocols for dogs after the diagnosis of secondary PH that might have impacted survival were not standardized. Our study had relatively strict enrollment criteria. We elected to follow recommendations from the ACVIM consensus guidelines and only enroll dogs with at least an intermediate probability of PH based solely on echocardiography,0.284), and right heart failure are associated with an increased risk of death in dogs with PH. Quantitative indices of right heart size and function provide clinically relevant prognostic information and are important in dogs with PH.In conclusion, our study suggests that increased right atrial size (quantified by RAA), RV systolic dysfunction (based on two‐dimensional echocardiography‐derived TAPSE < 3.23 mm/kg5Authors declare no conflict of interest.Authors declare no off‐label use of antimicrobials.Authors declare no IACUC or other approval was needed.Authors declare human ethics approval was not needed for this study.


Interactions between biomolecules control the processes of life in health and their malfunction in disease, making their characterization and quantification essential. Immobilization‐ and label‐free analytical techniques are desirable because of their simplicity and minimal invasiveness, but they struggle with quantifying tight interactions. Here, we show that mass photometry can accurately count, distinguish by molecular mass, and thereby reveal the relative abundances of different unlabelled biomolecules and their complexes in mixtures at the single‐molecule level. These measurements determine binding affinities over four orders of magnitude at equilibrium for both simple and complex stoichiometries within minutes, as well as the associated kinetics. These results introduce mass photometry as a rapid, simple and label‐free method for studying sub‐micromolar binding affinities, with potential for extension towards a universal approach for characterizing complex biomolecular interactions. Keeping count: Reported is the quantification of protein–protein interactions with light using mass photometry, which detects, images, and quantifies light scattering of single proteins upon binding to an illuminated glass‐water interface. The resulting signals are proportional to the molecular mass of each species and overall abundance, providing a new means to measure the relative abundances of proteins and their complexes, thereby enabling measurement of binding affinities and kinetics by molecular counting. Kd), free energies, and rate constants (koff and kon).Understanding how biomolecules interact with each other is central to the life sciences. The complexity thereof ranges from specific binary interactions, such as between antibodies and antigens,Kd<μm), which are often encountered for interactions of relevance for biopharmaceuticals in the context of antibody‐based drugs.In broad terms, existing biophysical methods can be categorized into size‐based approaches performing quantification and separation by either size or diffusion coefficient, physical interaction with functionalized surfaces, direct mass measurement, enthalpy changes, or light scattering.We have recently developed mass photometry (MP), originally introduced as interferometric scattering mass spectrometry (iSCAMS), as a means for detecting and measuring the mass of single proteins and the complexes they form in solution.Label‐free single‐molecule detection in principle provides the purest and most direct measurement of sample concentration by counting individual molecules. To explore this capability in the context of biomolecules, we chose monomers and domain exchanged dimers of the HIV‐1 neutralizing antibody 2G12 , which produced mass distributions with the expected major bands at 147 kDa and 291 kDa —(S6) and Figure S7]. IgG N‐glycan removal  weakened FcR bindingKd=1.0±0.1 nm .Equipped with these benchmarking results, we set out to investigate the suitability of MP to characterize interactions of varying affinities, using the immunoglobulin G (IgG) monoclonal antibody trastuzumab (Herceptin®) binding to soluble domains of IgG Fc receptors or ErbB2 (HER2) antigens. Trastuzumab, herein referred to as IgG, and FcγRIa by themselves revealed monodisperse distributions at 154±1 kDa and 50±1 kDa, respectively , suggesting that they were performed at or close to equilibrium . These binding affinities were confirmed after equilibration time screening .This simple single‐shot approach presented so far produces results in a few minutes, however, necessarily neglects the importance of kinetics and equilibration conditions. To address this, we probed FcγRIa binding to deglycosylated IgG at a 1:1 ratio. Samples were mixed at 4 μKd,1=1.4±0.1 nm and Kd,2=4.8±0.3 nm . Recording distributions at a few different concentrations, however, revealed a linear dependence of our Kd values on sample concentration, indicating a very tight Kd<70 pm, and/or slow interactions with off‐rates on the order of hours. Therefore, performing a few measurements at a range of concentrations is crucial to prevent misinterpreting data derived from a single‐shot Kd approach for very strong interactions. Irrespective, our method provides rapid and clear distinction between interactions with vastly different binding affinities, which only need to be refined if highly accurate measurements are required.For quantification of the tighter interaction between FcγRIa and IgG, screening at a range of concentrations was essential to ensure that the observed mass distributions were representative of the interaction to be quantified . As an additional example, for the HER2–IgG interaction, a simple single‐shot experiment at nanomolar concentration would have led to m concentration range, we should be able to access dissociation kinetics by simply diluting to total protein concentrations around the estimated Kd (approx. 1:1 ratio bound: unbound species for a 1:1 interaction), and monitoring the bound/unbound ratio throughout  than for deglycosylated IgG but an almost identical on‐rate , confirming that the glycans are critical for tight binding. Association measurements  can in principle be used in an analogous fashion, although we found it more susceptible to protein loss because of nonspecific adsorption . Overall, our results were in good agreement with SPR measurements association rates in addition to thermodynamic quantities raises the question to which degree we can use MP to directly visualize and quantify interaction kinetics. As MP measurements currently take place in the <100 nKd=31±11 nm . The resulting Kd values for the IgG‐FcRn interaction, at pH 5, were 44±9 nm for the monomer‐dimer equilibrium, 59±8 nm for the IgG‐FcRnmonomer, and 6.6±0.6 nm for IgG plus two FcRns [see Figure S30 and Equations (S7)–(S18)]. Increasing the pH value to 5.5 decreased the binding affinities to 171±19 nm and 225±20 nm but did not significantly affect the binding affinity of IgG plus two FcRns of 3.9±1.5 nm assembly completely label‐free and independent of protein immobilization to a surface, thus minimizing any possible perturbations, as well as being intrinsically sensitive to binding stoichiometries and oligomerization. The current limitation to sub‐micromolar affinities and concentration range can be addressed in the future through combination with fluidic approaches,m affinities, making MP a powerful approach for characterizing biomolecular interactions without labels and single‐molecule sensitivity in a minimally perturbative fashion. Furthermore, the applicability of MP to both nucleic acidsTaken together, we have demonstrated that molecular counting with MP is sensitive, quantitative, and accurate in determining the relative abundances of different biomolecules and their complexes in solution. When implemented in the vicinity of the binding affinity, a single measurement lasting typically 30 seconds, or 240 seconds for continuous flow injection, yields accurate binding affinities spanning four orders of magnitude from 30 pProtein preparation, mass photometry, data analysis as well as supplementary figures and equations are described in the Supporting Information.P.K. is an academic founder, shareholder and director to Refeyn Ltd. J.L.P.B. is an academic founder, shareholder and consultant to Refeyn LTd. W.B.S. is a shareholder and consultant to Refeyn Ltd. All other authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re‐organized for online delivery, but are not copy‐edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file.


Mindfulness training can enhance cognitive control, but the neural mechanisms underlying such enhancement in children are unknown. Here, we conducted a randomized controlled trial (RCT) with sixth graders (mean age 11.76 years) to examine the impact of 8 weeks of school‐based mindfulness training, relative to coding training as an active control, on sustained attention and associated resting‐state functional brain connectivity. At baseline, better performance on a sustained‐attention task correlated with greater anticorrelation between the default mode network (DMN) and right dorsolateral prefrontal cortex (DLPFC), a key node of the central executive network. Following the interventions, children in the mindfulness group preserved their sustained‐attention performance  and preserved DMN–DLPFC anticorrelation compared to children in the active control group, who exhibited declines in both sustained attention and DMN–DLPFC anticorrelation. Further, change in sustained‐attention performance correlated with change in DMN–DLPFC anticorrelation only within the mindfulness group. These findings provide the first causal link between mindfulness training and both sustained attention and associated neural plasticity. Administered as a part of sixth graders' school schedule, this RCT supports the beneficial effects of school‐based mindfulness training on cognitive control. Mindfulness training enhances cognitive control by preserving sustained‐attention performance  and preserving default mode network–dorsolateral prefrontal cortex (DMN–DLPFC) anticorrelation compared to children in the active control group, who exhibited declines in both sustained attention and DMN–DLPFC anticorrelation. Further, change in sustained‐attention performance correlated with change in DMN–DLPFC anticorrelation only within the mindfulness group. These findings provide the first causal link between mindfulness training and both sustained attention and associated neural plasticity. These processes include the ability to focus on a task for prolonged periods while inhibiting inappropriate responses or mind wandering  and the capacity to maintain goal‐relevant information in mind  . These dual processes correspond to brain activations in two neural networks: the central executive network (CEN) and the default mode network (DMN). The CEN, with core nodes located in bilateral dorsolateral prefrontal cortices (DLPFCs) and bilateral parietal cortices, typically exhibits increased activation during engagement in attention‐demanding tasks  between brain regions. The positive correlations are observed between brain regions that have often been identified as supporting orchestrated functions  on neurocognitive processes. The intervention was grade‐wide  and included 99 children (mean age 11.76 years). Results reported here come from the subset of children whose families opted to participate in a neuroimaging visit at pre‐ and post‐intervention, which was approximately one‐third (34.3%) of all children enrolled in the full‐scale RCT .In the present study, we measured sustained attention by performance on the Sustained Attention to Response Task (SART) before and after the interventions. This task requires the participant to press a button when presented with any digit , except for the rarely presented “3” that appears on only 5% of trials . As the task lasts approximately 15 min, it requires sustained attention for a tedious task over a long period. Performance on the Go trials provides a measure of sustained attention, whereas performance on the No‐Go trials provides a measure of response inhibition  program for low‐income families; and 10% were Hispanic, 32.5% were African American, 52.5% were White, and 5% other or multiple racial identities  behavioral assessments and imaging protocol Table . Thirty‐2.2A school‐based mindfulness training program was adapted by Calmer Choice  to be apEach class incorporated 5–15 min of mindfulness exercises requiring participants to attend to some aspect of present‐moment experience  and to refocus attention on the present moment when the mind engages with cognitive processes  or meta‐cognitive processes . Participants shared their experiences with the class and received personalized feedback from the instructor. Class content was designed to provide a clear set of strategies for practicing mindfulness as well as foster a conceptual understanding of mindfulness practice. Classes focused on (a) sitting in an upright posture with backs straight and gaze lowered or eyes closed, (b) distinguishing between naturally arising thoughts and elaborated thinking, (c) minimizing the distracting quality of past and future concerns by reframing them as mental projections occurring in the present, (d) using the breath as an anchor for attention during mindfulness exercises, (e) repeatedly counting up to five consecutive exhalations, and (f) allowing the mind to rest naturally rather than trying to suppress the occurrence of thoughts. The course lasted 8 weeks during which students met four times per week for 45‐min classes, totaling approximately 24 hr of group practice and instruction by the end of the intervention. The three trained instructors who led the intervention each had practical knowledge and experience in mindful awareness, as well as teaching mindfulness to children.2.3The SCRATCH  computerSCRATCH is a programming language and an online community where students program and share interactive media such as stories, games, and animations among them and with people from all over the world. Each class introduced step‐by‐step simple mathematical and computational ideas that were built into the SCRATCH experience in the first 15 min of the class. Students then applied the new knowledge to advance the creation of their individual programs in SCRATCH, thus applying core computational concepts such as iteration, conditionals, coordinates, variable random numbers, and so forth. Students were encouraged to share their experiences and their creative thoughts with the class and received personalized feedback from the instructor. Participants were also encouraged to work collaboratively and reason systematically. The two trained instructors who led the intervention each had practical knowledge and experience with the SCRATCH curriculum, and working with children.2.4We measured attentional characteristics through the SART  and “No‐Go” trials (targets). Accuracy on “Go” trials (hereafter: Go‐Accuracy) is an index of sustained attention  to ensure blinding of group assignment to testers. Students and teachers were instructed to not reveal group assignment. Pre‐ and post‐intervention MRI protocols were collected before and after the end of the intervention for all participants. Pre‐intervention MRI protocols were also administered before randomization to ensure blinding of group assignment to testers. MRI technicians and researchers were blind to group assignment at all times and participants were explicitly told not to reveal group assignment at any point.2.8Before the first MRI scan, all participants completed a mock‐scanner training session. Participants watched a cartoon movie in the mock scanner while their head motion was monitored. The movie would stop temporarily if their head moved more than 3 mm and resumed once no movement was detected. Recordings of the actual scanner sounds were played in the mock scanner during the training to acclimate participants to the scanner experience ahead of time. The mock scan session lasted about 30 min for each child.2.93; flip angle = 7°; TE = 1.61 ms; inversion time (TI) = 1,200 ms; and TR = 2,530 ms.At both neuroimaging sessions, participants underwent a 6‐min resting state scan where they were instructed to passively view a fixation cross during the scan period and not to close their eyes, sleep or engage in any mindfulness or other exercises for relaxation practices. Specific instructions were “Keep your eyes open, relax, try not to move and try to stay awake.” All scans were acquired using a 3 T Trio MR System with a 32‐channel, phased‐array head coil . Resting‐state functional magnetic resonance imaging (rs‐fMRI) was acquired using a gradient‐echo, echo‐planar imaging pulse sequence (EPI) with prospective acquisition correction (PACE) for motion  the overall rating of the class, (b) amount of work they had to do, (c) degree of active participation, and (d) how much practical knowledge they learned. Statistical significance level was set at .05.Statistical tests for student acceptability scores for both mindfulness and coding groups were conducted using R Studio version 1.0.136 with R version 3.6.0 . Denoising also included the regression of time points flagged as outliers due to motion, along with the seven realignment parameters  and their first‐order derivatives. In‐house custom software ART version 2015‐10 (Artifact Detection Tools (ART)  ) was use2.11.2a prior regions of interest (ROIs or nodes) in the DMN and CEN networks. Node regions for both networks were defined by the CONN software package and derived from an independent component analysis on 497 healthy control participants  as part of the Human Connectome Project (http://www.humanconnectome.org). DMN nodes . Significant main effects and interactions were followed up with post hoc testing. All analyses controlled for pre‐intervention performance, gender, and IQ to determine beta coefficients of the treatment effect. Unless otherwise stated all statistical analysis are nonparametric  with a height threshold of 33.1t(25) = 1.22, p = .23; mean and SD values for mindfulness group: 3.23 (1.23) and coding group: 2.7 (1.49)), the amount of work they had to do (t(25) = 1.60, p = .12; mindfulness group: 2.6 (1.12) and coding group: 3.3 (0.82)), degree of active participation (t(25) = .75, p = .45; mindfulness group: 3.1 (1.28) and coding group: 3.8 (.78)), and how much practical knowledge they learned (t(25) = 1.85, p = .08; mindfulness group: 2.6 (1.31) and coding group: 3.6 (.69)). These results show that there were no significant differences in student perceptions of the two training courses.Both groups reported similar acceptability for the mindfulness or coding training. There were no significant differences for the overall rating of the class (3.23.2.1t(30) = −7.45, p = 2.63e‐08, Cohen d = 1.89). The mindfulness‐training group exhibited significantly better Go‐Accuracy after the intervention compared to the coding‐training group  = 2.57, p = .01, Cohen f2= .47; Figure t tests between the groups showing no significant difference at pre‐intervention for Go‐Accuracy (t(30) = .21, p = .83), but significantly better Go‐Accuracy at post‐intervention (t(30) = 2.28, p = .01) in the mindfulness‐training group than the coding‐training group. Additionally, post hoc paired t tests revealed that children who received the mindfulness‐training intervention did not exhibit any significant pre–post intervention decline in Go‐Accuracy (t(14) = 1.50, p = .92), whereas children in the coding‐training group exhibited a significant pre–post decline in Go‐Accuracy (t(15) = 1.87, p = .04). There were no group differences in RT variability on Go trials both before the intervention (t(30) = .09, p = .92) or after the intervention  = .97, p = .33, Cohen f2= .01).At baseline, Go‐Accuracy was significantly less than 100% (t0 = −7.453.2.2t(30) = .91, p = .36) or after the intervention  = .12, p = .90, Cohen f2= .01) and on No‐Go‐RT‐variability before (t(30) = .05, p = .95) or after the intervention  = 1.53, p = .13, Cohen f2= .17).There were no significant group differences in No‐Go‐Accuracy before  = .72; p = .48), group  = .98; p = .33), and no significant interaction effects  = .84; p = .40) were observed . These results show that there were no significant group differences in motion. There was also no correlation between Motion and Go‐Accuracy .There were seven participants who exceeded the 20% movement threshold of images (or less than 132 usable time points) with movement outliers (4 mindfulness) and were discarded from the analysis. No significant effects of time .Across participants, prior to intervention, better Go‐Accuracy correlated with greater anticorrelation between DMN–CEN  = −3.35, p = .002, Cohen f2= .56, FWE‐corrected; Figure t tests between the groups showing no significant difference at pre‐intervention in DMN‐right DLPFC anticorrelation (t(30) = .07, p = .93), but significantly higher DMN‐right DLPFC anticorrelation at post‐intervention (t(30) = 2.35, p = .02) in the mindfulness‐training group than the coding‐training group. Additionally, post hoc paired t tests revealed that children in the mindfulness‐training group did not exhibit any significant post‐intervention decline in DMN‐right DLPFC anticorrelation (t(14) = .92, p = .81), whereas children in the coding‐training group exhibited a significant reduction in post‐intervention DMN–CEN anticorrelation (t(15) = 1.87, p = .04).The mindfulness‐training group showed a significant pre–post increase in DMN–CEN anticorrelation  after the intervention compared to the coding‐training group  = −3.69, p = .001, Cohen f2 = .59, FWE‐corrected). Furthermore, there was no significant difference between results with or without motion covariate as control variable .Given the importance of motion as a possible confound in functional connectivity, we performed analyses following Ciric et al.  to evalu3.3.4r = −.50, p = .03; coding training: r = .002, p = .50; Figure Only children in the mindfulness‐training group showed a significant and positive correlation between pre–post differences in Go‐Accuracy and pre–post differences in DMN‐right DLPFC anticorrelation  and the right DLPFC and right parietal components of the CEN (associated with cognitive control). This is the first evidence linking performance on an attention task with rsFC in children. Second, children who participated in an 8‐week long mindfulness training exhibited preserved sustained attention, whereas children in the active control condition exhibited a deterioration of sustained attention. Third, and mirroring the preservation of sustained attention, children who received mindfulness training exhibited preserved resting‐state anticorrelation between the DMN and right DLPFC, whereas children in the active control condition exhibited a deterioration of that anticorrelation. The relations among mindfulness training, sustained attention, and brain plasticity received further support by a correlation between changes in sustained attention performance and changes in DMN‐right DLPFC anticorrelation only in the children who received mindfulness training. Altogether, this study provides initial causal evidence for the brain basis of cognitive benefits due to mindfulness training in children.4.1The present findings provide initial evidence about how variation in sustained attention among children relates to variation in brain function. Prior to intervention and across all children, greater sustained attention was associated with greater resting‐state anticorrelation between the DMN and a major hub of the CEN, right DLPFC. This brain–behavior relation is consistent with prior findings in adults that the DMN and CEN play key roles in attentional processes and in individual differences in cognitive control. DMN–DLPFC anticorrelation is enhanced during attention‐demanding tasks  and multiple CEN regions in children ages 8–12, adolescents ages 13–17, and young adults ages 18–24  that supports sustained attention. In this case, mindfulness training may be seen as a protective factor against such neurocognitive effects of stress, as it is for behavioral effects of stress  in gender ratios between the two groups. However, we mitigated the effect of this imbalance by adding gender as a covariate in all analyses. Any additional characteristics that may have distinguished these families were equivalent across the two training groups.The study also has several strengths. First, it generalizes the benefits from mindfulness training beyond both active engagement in meditation and task‐specific brain plasticity. The changes in behavior and brain function occurred in a nonmeditative state. These findings are in agreement with the notion that mindfulness training transfers its effects to daily experiences beyond meditation practice  and a Quality Control measure (QC) indicative of average subject motion, before (top) and after (bottom) denoising of the BOLD signal.Figure S1.2. Distributions of functional connectivity values (bivariate correlation coefficients) in a network of 512 nodes across the entire brain, computed separately for each subject during pre‐ (top) and post‐ (bottom) sessions.Click here for additional data file.


Nigella sativa L., has attained attention by the researchers due to their pharmacologies perspectives such as prevention from cancer, antidiabetic and antiobesity, prevention from oxidative stress and cardioprotective disorder. Thymoquinone has been found to work as anticancer agent against different human and animal cancer stages including propagation, migration, and invasion. Thymoquinone as phytochemical also downregulated the Rac1 expression, mediated the miR‐34a upregulation, and increased the levels of miR‐34a through p53, as well as also regulated the pro‐ and antiapoptotic genes and decreased the phosphorylation of NF‐κB and IKKα/β. In addition, thymoquinone also lowered the metastasis and ERK1/2 and PI3K activities. The present review article has been piled by adapting narrative review method and highlights the diverse aspects of thymoquinone such as hepatoprotective, anti‐inflammatory, and antiaging through various pathways, and further utilization of this compound in diet has been proven effective against different types of cancers.The higher utilization of fruits and vegetables is well known to cure human maladies due to the presence of bioactive components. Among these compounds, thymoquinone, a monoterpene and significant constituent in the essential oil of  Thymoquinone is strong anticancer agent through multiple mechanistic routes. It also prevents the cardiotoxicity induced by doxorubicin through suppression of the carcinogenesis activity, membrane lipid peroxidation, destruction of the Fe‐NTA‐induced oxidative stress, and inhibition of the eicosanoid production  is the index protein of the brown adipose tissue. In recent study conducted by group of researchers, they found that thymoquinone treatment to the experimental subjects, which significantly increased the serum total antioxidant capacity, caused reduction in waist circumference, body weight, and body mass index  analogs have been found to improve the glycemic control, while administration of streptozotocin is used to induce the diabetes in experimental subjects. In diabetic rats, supplementation of thymoquinone by using oral route in dose‐dependent manner exhibited elevation in plasma GLP‐1 levels by lowering the hypergaphy , HDAC1,4,9, G9A, KMT2A,B,C,D,E, and KDM1B genes in Jurkat cells and MDA‐MB‐468 cancer cell lines  expression, inhibits interaction between HSP90 and HIF‐1α, and boosts HIF‐1α protein degradation in hypoxic cancer cells. It also suppresses the downstream genes of HIF‐1α, exhibited alterations in concentrations of lactate, glucose, and ATP levels, which further linked with disturbance of anaerobic metabolic and induction of apoptosis  dose‐dependently against MDA‐MB‐231 triple negative breast cancer in experimental mouse via inhibiting CXCR4 expression, tumor growth, and tumor vascularity along with suppressing the brain, lung, and bone metastases. Moreover, thymoquinone also suppresses the NF‐κB binding to the CXCR4 promoter and downregulation of the nuclear factor kappa‐light‐chain‐enhancer of activated B‐cell (NF‐κB) activation  against experimental subjects such as reduction in levels of glucose, glycosylated hemoglobin levels, urea and creatinine, NF‐κB levels, and liver enzyme concentrations , respectively  bioactive compounds to obese rats reverted these changes in the experimental rats  to the experimental subjects exhibited improvement in the glucose–insulin homeostasis‐related parameters, lipid profile parameters, integrity of pancreatic islets, and hepato‐renal functional and histomorphological statuses and enhanced the upregulated survivin, insulin‐producing β cells, endothelial cluster of differentiation 31, vascular endothelial growth factor, total glutathione, interleukin‐10 (IL‐10), and superoxide dismutase. Additionally, thymoquinone in the pancreatic tissues of STZ diabetic rats also downregulates the IL‐1β, caspase‐3, and TBARS levels  has been found to induce hepatotoxicity via depleting the concentration of glutathione enzyme, which further led to lipid peroxidation and subsequent liver injury. Thymoquinone in combination with curcumin prevented rats from liver injury through lowering the concentrations of alanine transaminase, total bilirubin, aspartate transaminase, and total protein. Additionally, reduction in iNOS and caspase 3 protein expressions and increment in the thioredoxin protein expression were also reported  and D‐galactose (60 mg kg−1 day−1) to experimental rats induced neurobehavioral and neuropathological alterations, whereas intragastrically administrated thymoquinone (20 mg kg−1 day−1) improved cognition, increased antioxidant enzymes level and B‐cell lymphoma‐2 levels, and decreased the acetylcholinesterase activities and nitric oxide in whole brain  to adult male Wistar Albino rats prevented rats from the cisplatin (15 mg/kg dose)‐induced myocardial injury, which further caused reduction in edema, congestion, and pycnotic nuclei in myocardial fibers, and enhancement in antiapoptotic protein Bcl‐2 level  injury through lowering infarct size, creatine kinase‐MB levels, and cardiac lactate dehydrogenase, promoting autophagy, improving cardiac function, and suppressing enedoxidative stress and apoptosis , eNOS, the components of the angiotensin system, and IK(Ca), and restoration of EDHF‐mediated relaxations and NO− in the mesenteric artery of middle‐aged rats  male mice  provides protection against intraperitoneally administrated cisplatin (12 mg/kg/body weight)‐induced hepatotoxicity via enhancing the antioxidant enzyme activities, that is, reduced glutathione contents, and decreasing MDA, TNF‐α, iNOS, and IL‐1β  to experimental mouse provide prevention against acetaminophen‐induced hepatotoxicity dose‐ and time‐dependently via lowering the serum alanine aminotransferase activities  phytochemical work as preventive agent when treated with interferon‐gamma (IFN‐γ)‐activated BV‐2 microglial cells and lipopolysaccharide (LPS) by momentously enhancing the expression of 4 antioxidant, neuroprotective proteins: glutaredoxin‐3, biliverdin reductase A, 3‐mercaptopyruvate sulfurtransferase, and mitochondrial ion protease, as well as also lowering the expression of inflammatory cytokines, IL‐6, IL‐2, IL‐4, IL‐10, and IL‐17a, respectively. Additionally, thymoquinone also downregulated the chemokine (CC) motif ligand 5, chemokine (CC motif) ligand 3, and complement factor B (CFB)  enhanced the neuron density in contralateral hippocampal regions  and lowered malondialdehyde level ‐induced toxicity in hippocampi of Wistar rats. It momentously decreased the mitochondrial dysfunction, mitochondrial membrane potential (Δψm), intracellular ROS generation, and apoptotic events  and thymoquinone  in combination with morphine (20 mg/kg) lowered the nitric oxide level, enhanced the motility , germinal thickness, morphology, count, viability of sperm cells, and testosterone hormone (Miah et al., 8.1In adjuvant‐induced arthritis, thymoquinone treatment provides protection against rheumatoid arthritis via reducing the expressions of IL‐1b and TNF‐a in adjuvant‐induced arthritis (Vaillancourt et al., 8.2Bronchial asthma is linked with airway inflammation and leukotrienes. Thymoquinone has been found to prevent from the deleterious effects induced by chemicals and environmental toxins. It also protects the lungs by exposing the toluene in rats Kanter, . Thymoqu9Nigella sativa. Evidently, thymoquinone is chemically known as 2‐methyl‐5‐isopropyl‐1, 4‐benzoquinone, an active principal component of the volatile oil that exhibits wide spectrum of health‐endorsing properties such as anti‐inflammatory, hepatoprotective, antimicrobial, antitumor, antimutagenic, antiepileptic, neuroprotective, and nephroprotective, respectively. Health‐associated perspectives of this bioactive compound led to medical applications. In several anti‐inflammatory and degenerative disorders such as cancer, thymoquinone also has been known to modify the multiple molecular and signaling pathways. Most important aspects of thymoquinone such as hepatoprotective, anti‐inflammatory, and antiaging have been highlighted through various pathways, and further utilization of this compound in diet has been proven effective against different types of cancers.Thymoquinone is a phytochemical compound found in the plant There is no conflict of interest among authors.Not Applicable.


Bone loss was also detected in cortical thickness and area fraction by day 21. However, four short bouts per week of compressive loading protected the loaded limb from much of this bone loss. At day 21, we observed a 50% loss in trabecular bone volume/total volume and a 6% loss in midshaft cortical thickness in unloaded limbs, but only 15% and 2% corresponding losses in contralateral loaded limbs (p = 0.001 and p = 0.02). Many bone geometry parameters of the loaded limbs of suspended animals did not significantly differ from non‐suspended control limbs. Conversely, this protective effect of loading was not detected in cortical bone at the lower‐strained distal shaft. Analysis of bone metabolism markers suggested that the benefits of loading occurred through increased formation instead of decreased resorption. This study uniquely isolates the role of externally applied mechanical loading of the mouse tibia, in the absence of muscle stimulation, in protecting bone from concurrent disuse‐related loss, and demonstrates that limited bouts of loading may be highly effective during prolonged disuse. © 2019 The Authors. JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.Prolonged reduction in weightbearing causes bone loss. Disuse of bone is associated with recovery from common musculoskeletal injury and trauma, bed rest resulting from various medical conditions, and spaceflight. The hindlimb‐suspension rodent model is popular for simulating unloading and disuse. We hypothesized that controlled mechanical loading of the tibia would protect against bone loss occurring from concurrent disuse. Additionally, we hypothesized that areas of high mechanical peak strains (midshaft) would provide more protection than areas of lower strain . Adult C57BL6/J mice were suspended for 3 weeks, with one limb subjected to tibial compression four times per week. μCT imaging was completed at days 0, 11, and 21, in addition to serum analysis. Significant bone loss caused by hindlimb suspension was detected in trabecular bone by day 11 and worsened by day 21 ( Prolonged disuse or limited weightbearing can cause significant detrimental effects on bone mass and strength. Disuse osteoporosis is a common skeletal disorder of the elderly and in patients subjected to prolonged immobility or bed rest, which can go unnoticed until it presents as fractures or spinal injuries.Therapeutic interventions, such as pharmacological, exercise, and nutritional strategies, have yielded variable success. Several countermeasures that have theoretical potential include use of prescription drugs  and training regimens that have a significant resistance‐exercise component.The hindlimb‐suspension unloading rodent model has been widely used for simulating unloading associated with spaceflight, and now is also used extensively to investigate muscle atrophy and disuse osteopenia caused by Earth‐based conditions such as muscle‐wasting disease, inactivity, bed rest, and immobilization.In vivo mechanical loading, creating higher strain environments, can induce an osteogenic atmosphere. in vivo loading models of turkey radius/ulna, rat femur/tibia, and murine femur/tibia have been reported.Although bone strain distributions resulting from tibial compression are complex, certain generalities can be made according to basic mechanics theory and findings from numerical modeling and experimental measurements: both traditional strain gauges and newer approaches such as digital image correlation.The differential between usual and novel strains has been reported to be most relevant to mechano‐responsiveness.The objective of this study was to examine the role of controlled mechanical loading in preventing bone loss associated with an otherwise catabolic state of bone disuse. It was hypothesized that limited bouts of mechanical loading would be protective from bone loss induced from hindlimb suspension, and secondarily that loading in cortical areas of high peak strain (midshaft) would provide protection, but loading in areas of lower strain  would not. Bone parameters were determined using μCT analysis and serum biomarkers. These hypotheses were tested by using the hindlimb‐suspension model in combination with the in vivo mechanical tibial‐loading model in C57Bl/6J mice. Although the hindlimb‐suspension and tibial‐compression models have been used individually, to our knowledge they have never been utilized simultaneously.ad libitum throughout the experiment. All mice in the facility were screened regularly by using a health‐monitoring program and were free from a wide range of pathogens. The mice were housed in a facility accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International. All animal use was performed according to the standards put forth in the Guide for the Care and Use of Laboratory Animals , and approved by the Penn State College of Medicine IACUC.Male WT C57BL/6J mice  were used. All mice were approximately 112 days (16 weeks) ± 3 days (25 g ± 2 g) at experimental outset, and therefore defined as skeletally mature at day 0 of the experiment. Mice were allocated randomly into experimental groups with internal controls for determining the primary effect of tibial compression . The same caging was used to house control mice.A modified model of hindlimb suspension was utilized, originally described by Morey‐Holton and Globus,On day 1 of the experiment, a noninvasive mechanical compression‐loading regime began on the right tibia of each mouse. Each mouse was loaded 4 days/week during the 21‐day experiment. The loading protocol was adapted from an osteogenic axial‐loading protocol described by Melville and colleagues.Mice were anesthetized with 1.5% isoflurane delivered in 100% oxygen for all μCT scans. Right and left tibias were scanned using a Scanco vivaCT 40 μCT  on day 0 as a baseline, day 11, and day 21 (at the study conclusion). Each limb was scanned in one 80‐min session. All scans were performed in vivo. Settings were 55 kVP, 145 μA, 200‐ms integration time. Image reconstruction was 2048 × 2048 matrices and isotropic voxels 10.4‐μm wide. Images were Gaussian‐filtered , and a 27.5% threshold was used to remove soft tissue. Trabecular sections from the proximal tibia, immediately distal to the growth plate, were evaluated over a 72‐slice region consistent with previous reports.Blood was collected on days 11 and 21 from two of the three groups of animals: HLS animals that were exposed to loading and the HLS only groups; blood was not collected from the control group. Approximately 250‐μL blood was collected using the submental or submandibular blood collection techniques at day 11. On day 21, blood was collected terminally from the heart after humane euthanasia. Blood samples were centrifuged, and serum was collected and stored at −80°C until serum analysis. Type I collagen N‐terminal propeptide (P1NP) and C‐terminal telopeptides (CTX) were quantified with ELISA kits  according to the manufacturer's protocols and using an Epoch Microplate Spectrophotometer . Outlier results outside the 95% confidence interval were removed.p < 0.05 was considered significant. The outcomes reported represent the percent change from baseline for each group of animals. For serum analysis, unpaired t tests were used to assess differences between groups.For the μCT data, each outcome was assessed using a general linear model with correlated errors that accounts for the longitudinal nature of the repeated measurements per animal. The general linear model with correlated errors uses restricted maximum likelihood and provides estimates of the least squares means, which differ slightly from raw observed means as the model takes into account such things as unequal group sample sizes. For each outcome, the Tukey–Kramer procedure was used to account for multiple comparisons testing between groups so that the overall family‐wise error rate was 0.05; hence, p = 0.17). For each time‐point, there were no significant differences between the groups.Throughout the experiment, the mean body weight loss for the hindlimb suspension (HLS) ± tibial compression (TC) group was −8% at day 21 compared with baseline; the mean body weight loss for the control group was −5% at day 21 compared with baseline (p < 0.001 at day 21), which averaged −1% and −12% at days 11 and 21, respectively.  Tibial compression of one limb of the hindlimb suspended animals (HLS + TC) prevented most of this disuse‐related bone loss, resulting in only −3% and −19% change in BV/TV at days 11 and 21, respectively. These losses were significantly lower at both time‐points (p = 0.001 at day 21) than the contralateral limb not receiving the tibial compression (HLS − TC). At day 21, the HLS + TC limb showed significantly higher loss in BV/TV (−19%) than the Control − TC limb (−12%), indicating that bone loss caused by disuse was not completely prevented by the periodic compression (p < 0.001). There was also a significant anabolic effect of tibial compression in the control animals (Control + TC versus Control − TC) at both time‐points .Representative 3D images of trabecular bone are shown in Fig. p < 0.05) at both time‐points and for all trabecular parameters including (in addition to BV/TV): Tb.Th (−21% at day 21 compared with baseline), Tb.N (−25%), Tb.Sp (+38%), BMD (−38%), Conn.D (−54%), and SMI (+70%). However, tibial compression of the hindlimb‐suspended animals (HLS + TC) prevented much of this loss, resulting in significant differences with the HLS − TC limb in all parameters at day 21 (p < 0.05), and in most parameters at day 11. For some parameters, there was no difference detected between the HLS + TC and Control − TC groups.In general, similar results were found for other trabecular parameters Fig. . HindlimA). The change on day 21 was significantly different than that of control mice (Control−TC limb), which had a mean gain of 1% (p = 0.002). Tibial compression of one limb of the hindlimb‐suspended animals (HLS + TC) prevented much of this disuse‐related bone loss, resulting in only −2% thickness at day 21, which was significantly less than the contralateral limb not receiving the tibial compression . Also, with respect to Ct.Th, at both days 11 and 21, the HLS + TC and Control − TC limbs were not significantly different, and the effects of tibial compression in the control animals were insignificant .Representative images of cortical bone are shown in Fig. A). For cortical midshaft μCT parameters, although significant effects of loading were observed in Ct.Th, it is notable that significant differences were not detected in cortical area, total area, or Ct.BV/TV . However, there were strong trends in the cortical area and Ct.BV/TV consistent with the Ct.Th result . Total area, Ma.Ar, and Ct.Po were not significantly affected by hindlimb suspension. Although there was no significance detected with Ma.Ar, there was substantial variability in the results. Baseline porosity measurements averaged 0.76% and at the final time‐point porosity averaged 0.88% in the groups analyzed. Although there were sizeable percent changes in porosity, the porosity values themselves were less than 1%.In general, similar trends were found for other cortical midshaft parameters Fig. A. For c/TV Fig. A. HowevB). This loss was significantly different than the change in control mice . Tibial compression of the loaded limb of the hindlimb‐suspended animals (HLS + TC) did not show significant differences compared with the unloaded contralateral limb (HLS − TC). There were also no significant effects of the tibial compression on thickness in the control animals .Hindlimb suspension (HLS−TC limb) induced a mean −1% and −3% loss in Ct.Th at the distal‐shaft location at days 11 and 21, respectively . Hindlimb suspension (HLS − TC) induced cortical bone loss with significant differences compared with the control limb (p < 0.05) at 21 days for the following parameters (in addition to thickness): cortical area (−3% at day 21 compared with baseline), total area (0%), and Ct.BV/TV (−3%). Ma.Ar and Ct.Po did not show significant effects of hindlimb suspension. At the distal location, tibial compression of the hindlimb‐suspended animals (HLS + TC) did not significantly affect any bone parameters compared with the unloaded limb (HLS − TC), and strong trends were not observed, except for total area .In general, similar trends were found for other cortical distal‐shaft parameters Fig. B. Hindlp = 0.009; Fig. At day 21, there was a significant increase in serum P1NP markers in the HLS + TC group (57.5 ng/mL) when compared with the separate group of HLS control animals , there were few significant differences detected between Control − TC and HLS + TC . There were no observed significant differences in any cortical parameters at day 11, and similar to trabecular bone, the mean changes in these samples were smaller at day 11 than at day 21.Previous experiments in C57Bl/6 mice that examined strain at midshaft showed that there was a general linear relationship between load and microstrain using load ranges of 3.8 to 11.6 N.At the midshaft location comparing the HLS groups, the Ct.Th increased with load, while the Ma.Ar also increased. This is consistent with a previous studyWe surmise that high peak strain at the trabecular and midshaft cortical locations in our experiment is creating the osteogenic effect. Schulte and colleagues demonstrated that local mechanical stimuli regulate bone formation and resorption at the tissue level in mice and that bone formation most likely occurs at sites of high local mechanical strain and resorption at sites of low mechanical strain.p < 0.0001) to our experiment with the suspension‐only group showing 16.95% lower BMD than the training‐during‐suspension group.p < 0.001) compared with the 20% increase that we report. They also report increases in Tb.Th (+58%), BV/TV (+85%), and Tb.N (+27%), compared with +20% Tb.Th, +32% BV/TV, and +10% Tb.N that we report with our experiment.Previous studies have reported that multiple methods of loading, exercise, and training are beneficial to protect bone from loss in the face of disuse. Experiments by Yanagihara and colleagues and Ju and colleagues demonstrated that continuous training in hindlimb‐suspended rats—in these cases, jumping—during a period of disuse is necessary to maintain bone quality.Serum biomarkers of bone formation and bone resorption may provide insight on the mechanism underlying observed tissue‐level changes. CTX is a common resorption marker of bone type I collagen by osteoclasts. Bone remodeling is a constant process that leads to an increase in circulation of type I collagen fragments associated with resorption. In a state of unloading or disuse, bone is in a constant state of resorption.Melville and colleagues have detailed the in vivo tibial axial loading model.Some important limitations exist in this study. Limitations include lack of measures of local/regional bone formation/resorption, eg, dynamic histomorphometry. Longitudinal in vivo μCT is limited to providing net 3D bone formation/resorption in each animal. For the μCT, a realignment of images to account for differences in limb alignment was not performed. The study tested a single tibial compression protocol, which may not be representative of other protocols. Strain levels were not experimentally measured in this study. Despite potential crosstalk between muscle and bone, this study was limited by focusing on only bone and not muscle, such as the tibialis anterior or gastrocnemius muscle. As a potential countermeasure to disuse, the loading regime focused on bone and may not have impacted muscle mass and strength loss substantially. The serum biomarkers are limited in that they are systemic, not local, in nature and could not be compared between loaded and contralateral control limbs.In conclusion, this study uniquely isolates the role of externally applied mechanical loading of mouse tibia, in the absence of muscle stimulation, in protecting bone from concurrent disuse‐related loss. Additionally, the study has fundamental implications for understanding the role of strain in the prevention of bone loss, especially in disuse scenarios.The authors declare that they have no conflicts of interest or disclosures in relation to the contents of this article.Fig. S1. Additional trabecular and cortical microstructural parameters obtained from μCT. Data shown as percent changes from baseline (day 0), with error bars indicating ± SEM. n = 10 to 15/group. Significant differences between groups within time‐points are indicated by assigned letters, p < 0.05. Significant differences between groups and baseline are indicated by *p < 0.05.Click here for additional data file.Table S1. Supplementary Tables.Click here for additional data file.


Low back pain is the most common spinal disorder among soldiers, and load carriage training (LCT) is considered the main cause. We aimed to investigate changes in the spine system of soldiers after LCT at high altitudes and the change trend of the lumbar spine and surrounding soft tissues under different load conditions.Magnetic resonance imaging scans of the lumbar spines of nine soldiers from plateau troops were collected and processed. We used ImageJ and Surgimap software to analyze changes in the lumbar paraspinal muscles, intervertebral discs (IVDs), intervertebral foramina, and curvature. Furthermore, the multiple linear regression equation for spine injury owing to LCT at high altitudes was established as the mathematical prediction model using SPSS Statistics version 23.0 software.2 to 9862.7 ± 456.4 mm2, and the functional CSA (FCSA) increased significantly from 8089.6 ± 707.7 mm2 to 8747.9 ± 426.2 mm2 after LCT (P < 0.05); however, the FCSA/CSA was not significantly different. Regarding IVD, the total lumbar spine showed a decreasing trend after LCT with a significant difference (P < 0.05). Regarding the lumbar intervertebral foramen, the percentage of the effective intervertebral foraminal area of L3/4 significantly decreased from 91.6 ± 2.0 to 88.1% ± 2.9% (P < 0.05). For curvature, the lumbosacral angle after LCT (32.4° ± 6.8°) was significantly higher (P < 0.05) than that before LCT (26.6° ± 5.3°), while the lumbar lordosis angle increased significantly from 24.0° ± 7.1° to 30.6° ± 7.4° (P < 0.05). The linear regression equation of the change rate, △FCSA% = − 0.718 + 23.085 × load weight, was successfully established as a prediction model of spinal injury after LCT at high altitudes.In the paraspinal muscles, the cross-sectional area (CSA) increased significantly from 9126.4 ± 691.6 mmThe spinal system encountered increased muscle volume, muscle congestion, tissue edema, IVD compression, decreased effective intervertebral foramen area, and increased lumbar curvature after LCT, which revealed important pathophysiological mechanisms of lumbar spinal disorders in soldiers following short-term and high-load weight training. The injury prediction model of the spinal system confirmed that a load weight < 60% of soldiers’ weight cannot cause acute pathological injury after short-term LCT, providing a reference supporting the formulation of the load weight standard for LCT. Chronic low back pain is the most common musculoskeletal disorder among soldiers. According to statistics, > 50% of soldiers have experienced low back pain during service , and theIn LCT, a soldier carries a certain weight of marching combat equipment according to the requirements of the mission, battlefield environment, weather conditions, and other factors when performing training or combat tasks. Then, the soldiers carry different weights across various terrains to complete various operational tasks to a high standard. A previous study described that under general temperature conditions when the marching speed is 5 km/h, the maximum allowable load weight for soldiers is 25 kg, and the most acceptable weight is 20 kg .At present, “plateau adaptive training” is an important training method for endurance and perseverance. In a high-altitude environment (> 500 m above sea level), some factors such as low partial pressure of air and oxygen caused by high altitude and dry and cold climate reduce the alveolar oxygen partial pressure, blood oxygen saturation, energy consumption, and metabolic rate of soldiers during training. Thus, both the maximum and acceptable load weights are reduced accordingly. Yin et al.  suggesteTo the best of our knowledge, there is a lack of standards for plateau LCT. Existing military studies on plateau training have focused more on respiration, sleep, or psychological problems of soldiers , whereasNine soldiers from combat troops in Xining City, Qinghai Province (2443 m above sea level), volunteered for this study. All subjects were of Han nationality with the following characteristics: age, 18–31 years; weight, 60–80 kg; height, 168–182 cm; and service life, 1–10 years on the plateau , and each subject provided written informed consent.Lumbar spine magnetic resonance imaging (MRI) was performed before and after the LCT by using the GE Signa Explorer 1.5 T MRI system at the Qinghai Red Cross Hospital. As for the examination time, some studies have shown variations on the lumbar flexion range, lumbar IVD height, and other biomechanical characteristics at different times of the day, known as the diurnal effect , 14. To Because of the limited supine position during training, we could not directly obtain actual spine images of soldiers who were standing with different load weights after training. Therefore, a pressure device was designed in advance for this study to simulate the soldiers’ loaded backs when they were in the lying position Fig. . DetaileAccording to the experimental design, to perform MRI before LCT, the subjects were lying supine on the device without support on the lumbar spine. Then, each subject carried a backpack weighting a percentage of their weight during LCT. After LCT, the subject was fixed to the device in the same posture, and the spring dynamometer was attached to one end of the rubber belt to apply a tension pressure, which is equal to the subject’s backpack weight, as the load weight. Finally, we affixed the rubber belt and performed MRI.2-weighted MRI scans. Because the boundary of the paraspinal muscle at the L3/4 vertebral level can be clearly identified, we used ImageJ version 1.51 software  to manually mark the contour of the paraspinal muscle at this level and then summed the CSAs of the following paraspinal muscles: bilateral erector spinae, bilateral psoas major, and bilateral quadratus lumborum , fat content, and proportions of fast-twitch fibers . The CSA2-weighted MRI scans was approximately 0–120 pixels (30–240 pixels for the bones and 120–660 pixels for the fats). Based on this fact, we obtained the gray values of images of the paraspinal muscles before and after LCT. Then, to determine the functional CSA (FCSA) of the paraspinal muscle, the threshold analysis method was used to select the image area in the CSA within the gray value threshold of 0–120 pixels. FCSA was used to represent the total content of lean muscle fibers within the boundaries of the lumbar paraspinal muscle fascia to measure changes in the muscle fibers. A related control study by Fortin et al. ..2-weightThe subjective questionnaire survey was performed and analyzed on nine participants. The questionnaire included basic information, degree of low back pain, degree of fatigue, and other physical symptoms. The low back pain level was rated by using an 11-point numerical rating scale (NRS) , and the degree of fatigue was self-rated from 0 to 10 , which means that the higher the score, the more the physical exertion , 27. AllP < 0.05) and simulation regression analysis were performed for the change rates (△Index%) of different indexes /pre LCT value). Detailed steps are as follows: 1) Shapiro-Wilk tests were used to check data normality (P > 0.05). 2) Paired sample t-tests were used to compare significant differences between each index before and after LCT (P < 0.05). 3) The main effect variance analysis method was used to analyze whether there were significant differences in the changes in various indicators between groups with different load weights and between those with different load times (P < 0.05). 4) Multiple comparison analysis was used to further compare differences in the index changes in the same load weight group or the same load time group (P < 0.05). 4) A specific index was selected to fit the multiple linear regression equation: the change rate before and after LCT = b1 + b2 × load weight + b3 × load time, which we used as a mathematical prediction model.All data of each research index were analyzed using SPSS Statistics version 23.0 . First, the mean value analysis was used to evaluate the characteristics of changes before and after LCT. Second, significance analysis . After the analysis of the main effect variance, no significant difference in the parameters of the lumbar paraspinal muscle between the two groups was found, regardless of whether they were grouped by different load weight or load time  and L4/5 level . A related study reported that the standard deviations of this measurement method between observers and within observers are 0.2 mm and 0.3 mm, respectively [The disc height at each level of Lectively . After t3/4 decreased from 91.6% ± 2.0 to 88.1% ± 2.9% after LCT, with an average reduction of 3.80%. The paired t-test indicated a significant difference in E-IF% of L3/4 . The E-IF% of L4/5 decreased from 89.6% ± 1.9 to 88.2% ± 1.7% after LCT, with an average reduction of 1.66%. The paired t-test indicated no significant difference in this index , while the LLA showed no significant difference  after LCT, while only one soldier suffered moderate back pain (NRS score 5). The fatigue degree score results indicated that all soldiers performed intense physical activities (fatigue degree score > 6) after LCT, and two of them felt totally exhausted (fatigue degree score 10) , indicating that the correlation is strong, and a linear relationship can be established. With regard to the load time, the correlation coefficients of △FCSA% was − 0.176, and the relative value was low (< 0.6), suggesting the absence of a linear relationship.F-value of this model was 9.612 with a P-value of 0.017 (< 0.1), indicating a linear relationship between △FCSA% and load weight. In the regression coefficient analysis, the constant term was − 0.718, and the partial regression coefficient b1 was 23.085 after the t-test (P = 0.017 (< 0.1)). The regression coefficient of load time outside the equation of the model was re-tested, and its P-value was 0.515 (> 0.1) which could not be introduced. The maximum absolute value of the standardized residual of this model was 1.422 (< 3). The normal p-p plot of regression standardized residual showed that the scattered points were basically distributed on the diagonal, which indicated that the residuals could be judged to be normally distributed  [Regarding the lumbar intervertebral foramen, the results showed that the E-IF% in the Ldiseases , 37. Maydiseases  suggeste = 0.56) . Our stuAs for the lumbar curvature, the LLA increased by 27.63%, while the LSA increased by 21.79% after LCT in the midsagittal image, both showing an increasing trend and having significant differences after LCT. Anatomically, the LLA and LSA can indirectly reflect the stability of the lumbar spine , 41. If In accordance with the questionnaire results, although we found that all soldiers suffered varying degrees of low back pain after LCT, most of them just experienced mild pain and only one soldier experienced moderate pain . These bThe linear regression equation of the change rate, △FCSA% = − 0.718 + 23.085 × load weight, was successfully established as a predictive model of soldiers’ spinal injury after plateau LCT. According to this equation, we believe that as the load weight increases, the △FCSA% will increase gradually, and the FCSA value will present an increasing trend. Therefore, we can use this prediction model to estimate that when the load weight reaches 100% of the soldier’s weight, the FCSA is expected to increase by approximately 22.367% after LCT. Since FCSA represents the lean muscle volume within the paraspinal muscle fascia, the variation trend of the prediction model further confirmed that plateau LCT can induce a certain proportion of hypertrophy and proliferation of muscle fiber cells. However, according to the physiological structure of the paraspinal muscle, the FCSA cannot increase indefinitely, and both the CSA and FCSA will eventually reach a peak with the increase in training intensity. Therefore, as the load weight continues to increase, we speculate that the partial regression coefficient b1 (23.085) of this prediction model will gradually decrease and the constant term (− 0.718) will gradually increase; finally, the △FCSA% will reach the upper limit, that is, the FCSA will reach the maximum threshold. However, this prediction model has not yet covered this range.According to the results of questionnaire survey and radiologists review, it can be concluded that when the maximum load weight does not exceed 60% of their weight, soldiers will not experience paraspinal muscle injury related to short-term LCT, proving that a load weight < 60% is within the tolerable load range of the lumbar paraspinal muscles. As a consequence, this prediction model can be applied to the current conventional weight-bearing training for soldiers in the plateau (the proportion of weight-bearing is less than 60%), which can provide a reference for the formulation of the load weight standard for plateau soldiers under training or combat and improve the scientificity and safety of LCT in the plateau environment.This study performed actual human experiments in plateau troops for LCT. Results should be interpreted in consideration of the limitations. First, due to military requirements and ethical principles, this experiment could not recruit a sufficient number of subjects. Although a uniform experimental design was used, it cannot completely offset the bias of test data and differences between subjects caused by the small sample size. We hope to recruit more subjects in subsequent studies to verify the present results and obtain further physiological changes in the spine system following plateau LCT. Second, due to ethical restrictions, we were unable to conduct LCT on soldiers using a load weight > 60% or a load time longer than 60 min at present; therefore, we could not directly determine the load weight and load time that may cause physiological damage. Thus, our prediction model also cannot cover the threshold range. We will further validate and optimize this model in subsequent experiments and estimate the training threshold through a more accurate mathematical model.This study directly confirmed that the spinal system encountered the following changes after short-term and high-load weight training: increased muscle volume, muscle congestion, tissue edema, IVD compression, decreased E-IF area, increased lumbar curvature, and other physiological changes that reveal important pathophysiological mechanisms of lumbar spinal disorders in soldiers. Moreover, this study established an injury prediction model of the spinal system for plateau LCT through multiple regression analysis: △FCSA% = − 0.718 + 23.085 × load weight. Combined with the results of the questionnaire survey and the radiologists review, this model confirmed that a load weight < 60% of soldiers’ weight cannot cause acute pathological injury in short-term LCT, providing a reference supporting the formulation of the load weight standard for the LCT and, thus, improving the scientificity and safety of military LCT in plateau environments. In addition, the simulated weight load device, image processing methods, and regression analysis method involved in this research have certain universality that can be applied to clinical research of spinal diseases and have considerable application prospects.


Several studies have argued that words evoking negative emotions, such as disgust, grab attention more than neutral words, and leave traces in memory that are more persistent. However, these conclusions are typically based on tasks requiring participants to process the semantic content of these words in a voluntarily manner. We sought to compare the involuntary attention grabbing power of disgusting and neutral words using them as rare and unexpected auditory distractors in a cross-modal oddball task, and then probing the participants’ memory for these stimuli in a surprise recognition task. Frequentist and Bayesian analyses converged to show that, compared to a standard tone, disgusting and neutral auditory words produced significant but equivalent levels of distraction in a visual categorization task, that they elicited comparable levels of memory discriminability in the incidental recognition task, and that the participants’ individual sensitivity to disgust did not influence the results. Our results suggest that distraction by unexpected words is not modulated by their emotional valence, at least when these words are task-irrelevant and are temporally and perceptually decoupled from the target stimuli. Numerous findings in psychology show that ongoing performance can be negatively affected by the presence of distracting stimuli, and that the degree of distraction exerted by such stimuli varies with certain factors. One prominent line of research has focused on the propensity of irrelevant stimuli to capture our attention by virtue of their unexpected occurrence and the contingent processing of their semantic features. Whether emotional features of such irrelevant stimuli can mediate distraction remains largely unexplored, however. A distinct line of work has centered on the role of the distractors’ emotional content, showing for example that distractors eliciting disgust are more likely to disrupt cognitive performance and imprint memory than neutral distractors. Though these findings are often interpreted as an indication of the negative distractors’ propensity to grab attention compared to neutral ones, they often originate from paradigms that were not designed to measure attention capture per se and require participants to attend the very stimuli conveying the emotional content. We sought to combine the two lines of work using a cross-modal oddball task designed to measure the distraction induced by to-be-ignored disgusting and neutral auditory words in an ongoing visual categorization task. We chose to focus on one specific negative emotion, namely disgust, for past work has suggested that it is especially prone to capture attention and imprint on memory.Task-irrelevant stimuli that unexpectedly differ from an otherwise structured or repeated sequence of stimuli (deviant among standard stimuli) capture attention and yield behavioral distraction in an unrelated ongoing task  of disgusting words  on distraction and memory measures, and setting the probability of Type I error to 0.05, the required sample size to achieve a power (1—type II error) of 0.95 is 45. Our sample size far exceeded this requirement.One hundred and twenty participants , with a mean age of 21.38 (SD = 4.77), took part in this study. All reported normal or corrected-to-normal vision and hearing. All were students at the University of the Balearic Islands and received a small honorarium for their participation. Informed consent was obtained from all individual participants included in the study. Under the hypothesis of a small-to-medium effect size . Participants were tested individually in a sound-attenuated testing booth. This study adhered to the ethical standards of the American Psychological Association, and received ethical approval from the Bioethics Committee of the University of the Balearic Islands.In this task, participants categorized visual stimuli while ignoring task-irrelevant sounds. Each trial consisted of the presentation of an irrelevant sound, immediately followed by the presentation of a visual target. Two categorization tasks were compared (between-participants). In the digit parity categorization task, participants categorized visually presented digits (1–4) as odd or even. In the semantic categorization task, participants categorized visually presented words as musical instruments or vehicles. In both task conditions, participants were presented with 720 trials . Each trial consisted of the following sequence of events. A white fixation cross appeared at the center of a black screen, accompanied by a task-irrelevant sound (described below). The fixation cross and the sound started at the same time and lasted 400 ms. The fixation cross was then replaced by the visual target stimulus (presented in white) during 200 ms, after which the fixation cross returned during 800 ms. Participants, therefore, had a total response window of 1000 ms from the target’s onset. The next trial started automatically at the end of this interval. Participants responded by pressing the B and N keys on the computer keyboard, using two fingers of their dominant hand. The mapping between the response keys and the odd/even or instrument/vehicle responses  was counterbalanced across participants.Three types of sound  were mixed in a quasi-random order of presentation within each block of trials. In standard trials , the sound consisted of a 650 Hz sine wave tone, hereafter referred to as the standard sound. In disgusting deviant trials , the sounds consisted of the audio recordings from one of two sets of 12 words evoking disgust . In neutral deviant trials , we used audio recordings of 12 neutral words . Each of the 24 deviant words  were used once within each block . All sounds were presented binaurally through headphones at an intensity of approximately 70 dB SPL. A different quasi-random order of presentation of the standard, disgusting and neutral trials was used for every participant, with the constraint that deviant trials never occurred on consecutive trials, and that each type of sound trial occurred equally often in conjunction with each of the visual target stimuli.In the digit parity task, the target stimuli consisted of the digits 1–4, each occurring equally often in each block of trials. In the semantic judgement task, six words corresponded to musical instruments and six words corresponded to vehicles. Each word was used equally often in each block of trials.Each block of test trials was preceded by warmup trials involving the standard sound only: 8 practice trials in the digit parity task (digits 1–4 presented twice each), and 12 practice trials in the semantic categorization task . Participants were instructed to concentrate on the categorization of the visual stimuli, to try to respond as quickly as possible while trying not to make errors, and to ignore all sounds.Following the oddball task, participants performed a surprise recognition task to measure the extent to which they recognized the auditory words used as deviant stimuli in the oddball task. In each of 48 trials, participants were presented with an auditory word and asked to indicate whether they recognized that word as one of those presented in the oddball task. Participants responded by pressing the numerical keys “1” and “2” using two fingers of their dominant hand . Following the presentation of each word and until a response was registered; graphical illustrations of the 1 and 2 keys were visible on the left and right sides of the screen respectively, accompanied by the words “yes” and “no”. Each participant was presented with 24 disgusting and 24 neutral words . The sets of 12 words were rotated across participants. The order of the words was random and different for every participant. Participants were instructed to respond as quickly as possible while trying to make no error.α = 0.825).Following the administration of the recognition task, each participant completed the Spanish version , to assess the credibility of the experimental hypothesis (presence of an effect) relative to that of the null hypothesis (absence of an effect) given the data. While values below 1 indicate that the null hypothesis is more credible than the experimental hypothesis , it is often considered that values below 1/3 are considered as strong support for the null effect, while values above 3 are regarded as strongly supporting the presence of an effect , the decision criterion (C) and RTs in the recognition task  and task type  on response times (RTs) and the proportion of correct responses in the cross-oddball task, and on the sensitivity index (https://osf.io/8tmez).All the analyses were conducted using JASP  = 49.363, MSE = 423.4, p < 0.001, BF10 = 3.942 × 1015. Further analysis revealed that both deviant words produced significant distraction relative to the standard sound (disgusting deviant words vs standard: t(119) = 8.898, dz = 0.812, 95% CI 0.605–1.017, p < 0.001, BF10 = 8.754 × 1011; neutral deviant words vs standard: t(119) = 7.300, dz = 0.666, 95% CI 0.467–0.863, p < 0.001, BF10 = 2.417 × 108). In contrast, no difference was found between the two types of deviant words, t(119) = 1.066, dz = 0.097, 95% CI − 0.082 to 0.276, p = 0.288, BF10 = 0.176.Separate one-way ANOVAs for repeated measures were carried out to examine the effect of the type of sound trial  on RTs (ms) and on the proportion of correct responses see Fig. . The anaF = 0.674, MSE = 0.003, p = 0.511, BF10 = 0.056. A planned contrast aimed at comparing the two types of deviant words confirmed the absence of any difference between these conditions in that respect, t(119) = 0.049, ΔM = 1.986 × 10−4, dz = 0.004, 95% CI − 0.174 to 0.183, p = 0.961, BF10 = 0.102.The type of sound trial did not affect the proportion of correct responses, r = 0.123, 95% CI − 0.057 to 0.296, p = 0.179, BF10 = 0.279), or for the proportion of correct responses .To determine whether distraction by disgusting deviant words varied with the participants’ sensitivity to disgust, we calculated the correlation between, on the one hand, the difference between the two types of deviant word  and, on the other hand, the participants’ score on the disgust sensitivity questionnaire. No correlation was found for RTs , or for the proportion of correct responses .Finally, we examined the relationship between distraction due to acoustic deviations (standard versus deviant words) and distraction due to the meaning of the deviant words  by analyzing the correlation between these two measures. No significant correlation was found for RTs  and the decision criterion (C), as well as response times.Performance in the recognition task was measured and analyzed using two measures based on signal detection theory, namely the sensitivity index (d′) for each of the two types of deviant word . As visible in Fig. t(119) = 0.745, dz = 0.068, 95% CI − 0.111 to 0.247, p = 0.458, BF10 = 0.133. Furthermore, no correlation was observed between the difference in d′ between the two types of deviant words and the participant’s sensitivity to disgust . In sum, both frequentist and Bayesian statistics revealed that participants recognized disgusting and neutral deviant words equally well, irrespective of their sensitivity to disgust.We calculated the sensitivity index (t(119) = 11.919, dz = 1.088, 95% CI 0.861–1.313, p < 0.001, BF10 = 9.157 × 1018  did not correlate with the participants’ sensitivity to disgust . The results indicate that participants adopted a more liberal decision criterion for disgusting word than for neutral words, which was not related to their sensitivity to disgust.The decision criterion (C) was significantly lower for disgusting deviant words than for neutral deviant words, indicating that participants were more inclined to respond “yes” to disgusting probes, see Fig. b. This dF = 1.168, MSE = 20,527.121, p = 0.282, BF10 = 0.161), nor that of the type of probe  = 0.108, MSE = 29,196.523, p = 0.743, BF10 = 0.107), were statistically significant. However, the sound type × probe type interaction was significant  = 13.329, MSE = 24,285.124, p < 0.001, BF10 = 63.941). Follow-up tests revealed that disgusting words yielded significantly slower RTs than neutral words for negative probes (t(119) = 2.707, ΔM = 37.803, dz = 0.247, 95% CI 0.065–0.428, p = 0.008, BF10 = 3.284), while the reverse was observed for positive probes (t(119) = − 2.812, dz = − 0.257, 95% CI − 0.438 to − 0.074, p = 0.006, BF10 = 4.288). These results are illustrated in Fig. r = −0.074, 95% CI − 0.250 to 0.107, p = 0.424, BF10 = 0.157: positive probes: r = − 0.095, 95% CI − 0.270 to 0.085, p = 0.300, BF10 = 0.194). This pattern of response times is compatible with the finding that disgusting words bias decisions toward a positive response. For positive probes, this speeds up responses to disgusting words. However, for negative probes, this bias must be cancelled out to select a negative response, thereby lengthening RTs for disgusting words relative to neutral words.Mean response times (RTs) were analyzed using a 2  × 2  ANOVA for repeated measures. Neither the main effect of sound type  and response times (RTs) in the recognition task. These findings fit with the finding that negative and arousing stimuli bias decisions toward the “yes” response in recognition tasks . Interestingly, while Ljungberg and Parmentier  observedIn summary, our results suggest that disgusting words do not grab attention more than neutral words in a cross-modal oddball task, at least as can be measured at a behavioral level. The notion that disgusting words are better remembered because they grab attention more than neutral words was not supported in our study, at least as we tested it . Taking into consideration previous studies, we hypothesize that a memory advantage for disgusting over neutral words might only manifest itself in tasks where participants voluntarily attend these words. Finally, we suggest that further work is necessary to explore whether disgust may modulate distraction in the cross-modal oddball task at the electrophysiological level or when it is conveyed through prosody.