Mutational analysis in BCR-ABL1 positive leukemia by deep sequencing based on nanopore MinION technology.

We report a third-generation sequencing assay on nanopore technology (MinION) for detecting BCR-ABL1 KD mutations and compare the results to a Sanger sequencing(SS)-based test in 24 Philadelphia-positive (Ph+) leukemia cases. Our data indicates that MinION is markedly superior to SS in terms of sensitivity, costs and timesaving, and has the added advantage of determining the clonal configuration of multiple mutations. We demonstrate that MinION is suitable for employment in the hematology laboratory for detecting BCR-ABL1 KD mutation in Ph+ leukemias.

Myelodysplastic syndrome with 5q deletion following IgM monoclonal gammopathy, showing gene mutation MYD88 L265P.

Patients affected by monoclonal gammopathy of undetermined significance (MGUS) very rarely develop a myelodysplastic syndrome (MDS). However, it was also demonstrated that MGUS patients had a significantly increased risk of developing MDS compared to the general population. We report a case of 5q-syndrome following a MGUS IgMk with mutation of MYD88 L256P. To our knowledge, this is the first case of del(5q) MDS following MGUS IgMk with the MYD88 L256P mutation in which there is coexistence of the markers of the two clonal diseases, but as an expression of distinct pathological features.

ADAMTS2 gene dysregulation in T/myeloid mixed phenotype acute leukemia.

BACKGROUND: Mixed phenotype acute leukemias (MPAL) include acute leukemias with blasts that express antigens of more than one lineage, with no clear evidence of myeloid or lymphoid lineage differentiation. T/myeloid (T/My) MPAL not otherwise specified (NOS) is a rare leukemia that expresses both T and myeloid antigens, accounting for less than 1% of all leukemias but 89% of T/My MPAL. From a molecular point of view, very limited data are available on T/My MPAL NOS. CASE PRESENTATION: In this report we describe a T/My MPAL NOS case with a complex rearrangement involving chromosomes 5 and 14, resulting in overexpression of the ADAM metallopeptidase with thrombospondin type 1 motif, 2 (ADAMTS2) gene due to its juxtaposition to the T cell receptor delta (TRD) gene segment. CONCLUSION: Detailed molecular cytogenetic characterization of the complex rearrangement in the reported T/My MPAL case allowed us to observe ADAMTS2 gene overexpression, identifying a molecular marker that may be useful for monitoring minimal residual disease. To our knowledge, this is the first evidence of gene dysregulation due to a chromosomal rearrangement in T/My MPAL NOS.

Statistical Parametric Mapping to detect the effects of the secondary jump direction on landing kinematics.

Anterior cruciate ligament (ACL) injuries in sports often occur with non-contact mechanisms, such as landing and cutting. Previous studies explored the ACL injury biomechanical risk factors through drop-jumps combined with secondary jumps. This study aimed to investigate the effect of the secondary jump direction on first landing kinematic temporal series. Fifty-seven participants (29 males, 28 females) performed three single-leg drop-jumps followed by secondary jumps in vertical (single-planar), 45°-medial and 45°-lateral direction (multi-planar). Lower limb and trunk landing kinematics was recorded using a 9-camera motion capture system and analysed with a One-way ANOVA through Statistical Parametric Mapping (SPM), from initial contact to maximum knee flexion. All variables were affected by the secondary jump direction, except trunk rotation. In sagittal plane, kinematic main differences were found between single- and multi-planar tasks. The latter elicited higher trunk, hip, and knee flexion. Frontal plane kinematics was more influenced by medio-lateral components of secondary jumps. Our results could underline how a single task may be insufficient for ACL injury risk assessment. Single- and multi-planar tasks including a secondary jump should be considered for more comprehensive evaluations in prevention and rehabilitation programs, but caution should be used when comparing results of studies adopting different tasks.

Customized Landing Task for ACL Injury Risk Assessment: Kinematic Sex-Related Differences.

BACKGROUND: Women present a higher anterior cruciate ligament (ACL) injury rate than men, suggesting sex-related biomechanical differences. Task characteristics are often fixed for both sexes, possibly affecting the perceived difficulty. We investigated kinematic sex differences across landing tasks for ACL injury risk assessment, adjusted to participants' anthropometrics/performance, and whether different tasks affect kinematic sex comparisons. HYPOTHESIS: Female subjects would exhibit motion patterns more associated with ACL injury risk, and sex-related differences may depend on task type. STUDY DESIGN: Descriptive laboratory study. LEVEL OF EVIDENCE: Level 3. METHODS: A total of 27 female and 29 male amateur players (18-30 years) executed horizontal hop, drop jump (DJ), and DJ followed by vertical or forward jump (length, proportional to maximal forward jump; box, 20% participant's height). An optoelectronic system provided lower limb kinematics at initial contact and peaks until maximum knee flexion (KF), analyzed separately by multivariate analysis of variance (MANOVA) (α = 0.05). RESULTS: At initial contact, the interaction term had significant effects on hip adduction (P < 0.01) and knee abduction (KAb) (P = 0.04); female participants demonstrated higher KAb (P < 0.01) and knee internal rotation (P = 0.05). For peaks analysis, the interaction term had no significant effects on any individual variable, although significant in MANOVA; female participants had higher KAb (P = 0.01) and lower KF (P = 0.04). Task type affected hip flexion and knee angles in both analyses. CONCLUSION: All variables in which significant sex-related differences were found are potential ACL injury risk factors, and all findings indicate that the analyzed female sample exhibited higher injury-related patterns. Although customized, male and female participants showed different landing strategies depending on the task. CLINICAL RELEVANCE: The findings underline how female participants adopted potentially harmful kinematics while executing customized landing tasks (adjusted by subject's anthropometrics/performance), which may enhance risk of ACL injury.

Soccer and volleyball players do not land differently: implications for anterior cruciate ligament injury risk.

BACKGROUND: Non-contact anterior cruciate ligament injuries are common in soccer and volleyball, occurring during changes of direction and landings. This study aimed to investigate kinematic differences between soccer and volleyball players in single-planar and multiplanar landing tasks, simulating sport-specific injury mechanisms. Since the anterior cruciate ligament injury rate in soccer is higher than in volleyball, we hypothesized that volleyball players would adopt safer landing strategies, especially in single-planar landing tasks. METHODS: Twenty-two soccer and 19 volleyball players performed single-leg drop landing, drop jump in vertical, 45°-medial and 45°-lateral directions. Box height and jump length were adapted to the subject's height and performance level, respectively. A 9-camera motion capture system provided lower limb kinematics. Two mixed multivariate analyses of covariance (sport, task, sex as covariate) were used to compare soccer and volleyball players' initial contact and peak kinematics (α=0.05). RESULTS: Task had significant effects on lower limb initial contact and peak angles, as expected. Sport and task × sport interaction had no significant effects on kinematics. CONCLUSIONS: Soccer and volleyball players' landing strategies were thus similar in each task, in opposition to initial hypotheses. We might speculate that the higher anterior cruciate ligament injury rate in soccer may be more related to non-predictable factors than the isolated landing kinematics.

Effects of mediolateral whole-body vibration during gait with additional cognitive load.

Whole-body vibration (WBV) may increase musculoskeletal disorder risk among workers standing on vibrating surfaces for prolonged periods. Limited studies were conducted to comprehend WBV impact on individuals engaged in dynamic activities. This study explored the effects of different horizontal WBV frequencies on gait parameters, lower limb kinematics, and the cognitive response of healthy subjects. Forty participants walked at constant speed on a treadmill mounted on a horizontal shaker providing harmonic vibration with an amplitude of 1 m/s2 and frequencies 2-10 Hz, with inversely proportional amplitudes. A Psychomotor Vigilance Test measured reaction time while a motion capture system recorded walking kinematics. ANOVA results revealed no significant impact of vibration frequencies on the reaction time. At 2 Hz, alterations in gait spatiotemporal parameters were significant, with reduced stride length, stride time, step length, and stance time and increased step width and cadence. Similarly, gait variability measured by standard deviation and coefficient of variation significantly increased at 2 Hz compared to the other conditions. Comparably, kinematic time series analyzed through statistical parametric mapping showed significant adjustments in different portions of the gait cycle at 2 Hz, including increased hip abduction and flexion, greater knee flexion around the heel strike, and augmented ankle dorsiflexion. Participants exhibited gait kinematic variations, mainly at 2 Hz, where the associated mediolateral displacement was higher, as a plausible strategy to maintain stability and postural control during perturbed locomotion. These findings highlight individuals' complex biomechanical adaptations in response to horizontal WBV, especially at lower frequencies, under dual-task conditions.

Clonal B cells in patients with hepatitis C virus-associated mixed cryoglobulinemia contain an expanded anergic CD21low B-cell subset.

Hepatitis C virus (HCV) is associated with the B-cell lymphoproliferative disorders mixed cryoglobulinemia (MC) and non-Hodgkin lymphoma. We have previously reported that HCV(+)MC(+) patients have clonal expansions of hypermutated, rheumatoid factor-bearing marginal zone-like IgM(+)CD27(+) peripheral B cells using the V(H)1-69 gene. Here we coupled transcriptional profiling with immunophenotypic and functional studies to ascertain these cells' role in MC pathogenesis. Despite their fundamental role in MC disease, these B cells have overall transcriptional features of anergy and apoptosis instead of neoplastic transformation. Highly up-regulated genes include SOX5, CD11C, galectin-1, and FGR, similar to a previously described FCRL4(+) memory B-cell subset and to an "exhausted," anergic CD21(low) memory B-cell subset in HIV(+) patients. Moreover, HCV(+)MC(+) patients' clonal peripheral B cells are enriched with CD21(low), CD11c(+), FCRL4(high), IL-4R(low) memory B cells. In contrast to the functional, rheumatoid factor-secreting CD27(+)CD21(high) subset, the CD27(+)CD21(low) subpopulation exhibits decreased calcium mobilization and does not efficiently differentiate into rheumatoid factor-secreting plasmablasts, suggesting that a large proportion of HCV(+)MC(+) patients' clonally expanded peripheral B cells is prone to anergy and/or apoptosis. Down-regulation of multiple activation pathways may represent a homeostatic mechanism attenuating otherwise uncontrolled stimulation of circulating HCV-containing immune complexes.

Head-centric computing for vestibular stimulation under head-free conditions.

Background: The vestibular end organs (semicircular canals, saccule and utricle) monitor head orientation and motion. Vestibular stimulation by means of controlled translations, rotations or tilts of the head represents a routine manoeuvre to test the vestibular apparatus in a laboratory or clinical setting. In diagnostics, it is used to assess oculomotor postural or perceptual responses, whose abnormalities can reveal subclinical vestibular dysfunctions due to pathology, aging or drugs. Objective: The assessment of the vestibular function requires the alignment of the motion stimuli as close as possible with reference axes of the head, for instance the cardinal axes naso-occipital, interaural, cranio-caudal. This is often achieved by using a head restraint, such as a helmet or strap holding the head tightly in a predefined posture that guarantees the alignment described above. However, such restraints may be quite uncomfortable, especially for elderly or claustrophobic patients. Moreover, it might be desirable to test the vestibular function under the more natural conditions in which the head is free to move, as when subjects are tracking a visual target or they are standing erect on the moving platform. Here, we document algorithms that allow delivering motion stimuli aligned with head-fixed axes under head-free conditions. Methods: We implemented and validated these algorithms using a MOOG-6DOF motion platform in two different conditions. 1) The participant kept the head in a resting, fully unrestrained posture, while inter-aural, naso-occipital or cranio-caudal translations were applied. 2) The participant moved the head continuously while a naso-occipital translation was applied. Head and platform motion were monitored in real-time using Vicon. Results: The results for both conditions showed excellent agreement between the theoretical spatio-temporal profile of the motion stimuli and the corresponding profile of actual motion as measured in real-time. Conclusion: We propose our approach as a safe, non-intrusive method to test the vestibular system under the natural head-free conditions required by the experiential perspective of the patients.

The Effects of FIFA 11+ Kids Prevention Program on Kinematic Risk Factors for ACL Injury in Preadolescent Female Soccer Players: A Randomized Controlled Trial.

This study aimed to investigate the effects of the 8-week FIFA 11+ Kids program on kinematic risk factors for ACL injury in preadolescent female soccer players during single-leg drop landing. For this, 36 preadolescent female soccer players (10-12 years old) were randomly allocated to the FIFA 11+ Kids program and control groups (18 players per group). The intervention group performed the FIFA 11+ Kids warm-up program twice per week for 8 weeks, while the control group continued with regular warm-up. Trunk, hip, and knee peak angles (from initial ground contact to peak knee flexion) were collected during the single-leg drop landing using a 3D motion capture system. A repeated measure ANOVA was used to analyze groups over time. Significant group × time interactions were found for the peak knee flexion, with a medium effect size (p = 0.05; effect size = 0.11), and peak hip internal rotation angles, with a large effect size (p < 0.01; effect size = 0.28). We found that the FIFA 11+ Kids program was effective in improving knee flexion and hip internal rotation, likely resulting in reducing ACL stress during single-leg drop landing in young soccer players.

Animal conflicts escalate in a warmer world.

The potential for climate change to affect animal behaviour is widely recognized, yet its possible consequences on aggressiveness are still unclear. If warming and drought limit the availability of food resources, climate change may elicit an increase of intraspecific conflicts stemming from resource competition. By measuring aggressivity indices in a group-living, herbivorous mammal (the Apennine chamois Rupicapra pyrenaica ornata) in two sites differing in habitat quality, and coupling them with estimates of plant productivity, we investigated whether harsh climatic conditions accumulated during the growing season influenced agonistic contests at feeding via vegetation-mediated effects, and their interaction with the site-specific habitat quality. We focused on females, which exhibit intra-group contest competition to access nutritious food patches. Accounting for confounding variables, we found that (1) the aggression rate between foraging individuals increased with the warming accumulated over previous weeks; (2) the probability to deliver more aggressive behaviour patterns toward contestants increased with decreasing rainfall recorded in previous weeks; (3) the effects of cumulative warming and drought on aggressivity indices occurred at time windows spanning 15-30 days, matching those found on vegetation productivity; (4) the effects of unfavourable climatic conditions via vegetation growth on aggressivity were independent of the site-specific habitat quality. Simulations conducted on our model species predict a ~50 % increase in aggression rate following the warming projected over the next 60 years. Where primary productivity will be impacted by warming and drought, our findings suggest that the anticipated climate change scenarios may trigger bottom-up consequences on intraspecific animal conflicts. This study opens the doors for a better understanding of the multifactorial origin of aggression in group-living foragers, emphasising how the escalation of agonistic contests could emerge as a novel response of animal societies to ongoing global warming.

Alterations in the antigen processing-presenting machinery of transformed plasma cells are associated with reduced recognition by CD8+ T cells and characterize the progression of MGUS to multiple myeloma.

We hypothesized that progression of monoclonal gammopathy of undetermined significance (MGUS) to multiple myeloma (MM) reflects the escape of transformed plasma cells from T-cell recognition because of impaired antigen processing-presenting machinery (APM). We studied plasma cells and CD8(+) T cells from bone marrow of 20 MGUS patients, 20 MM patients, and 10 control patients. Immunofluorescence and flow cytometry revealed significantly different patterns of APM component expression in plasma cells from the 3 groups. Compared with control patients, MM samples had lower expression of proteasome subunits and peptide transporters and greater expression of chaperones, considering both percentages of stained cells and molecular equivalents of soluble fluorochrome. MGUS samples had intermediate percentages of stained cells but molecular equivalents of soluble fluorochrome similar to control patients. Real-time polymerase chain reaction documented that APM changes occurred at the transcriptional level. Cytotoxicity assays demonstrated that MGUS CD8(+) T cells lysed autologous transformed plasma cells more than MM CD8(+) T cells did. MGUS progression correlated directly with calnexin, calreticulin, and tapasin and indirectly with delta, LMP2, and LMP10 expression levels; MM disease status did not correlate with APM levels. APM changes may allow transformed plasma cells to elude immunesurveillance in the MGUS-MM pathogenetic sequence.

The influence of task type and movement speed on lower limb kinematics during single-leg tasks.

BACKGROUND: Single-leg squats and step-downs are commonly used to assess kinematic variables that may be linked to injuries. Task type and movement speed may influence the outcomes of interest because of different balance requirements. RESEARCH QUESTION: What is the influence of task type and movement speed on lower limb kinematics? METHODS: This is a cross-sectional within-subjects study where 22 physically active females performed three single-leg functional tasks (Squat, Anterior step-down, and Lateral step-down) at three movement speeds (slow [5 s], fast [2 s], and self-selected), while three-dimensional kinematic variables were recorded. Displacement values from the initial position in single-leg support until 60° or peak knee flexion were calculated. Two-way repeated measures ANOVA was used to compare tasks and speeds, and Cohen's d effect size (ES) was calculated for significant pairwise comparisons. RESULTS: At 60°, lateral step-down presented the greatest hip adduction (large ES) and internal rotation (small ES). The anterior step-down had the lowest knee abduction displacement while the squat had the greatest (small to medium ES). At peak knee flexion, values increased but differences between tasks followed a similar pattern. Slow speed induced smaller displacement angles at the knee and hip (trivial to small ES). SIGNIFICANCE: When knee abduction is the variable of interest, the SLS may be the best test since it elicits the greatest displacement, but when evaluating hip motion, SDLAT might be best. Knee abduction and internal rotation were lowest in the slow condition, suggesting that faster speed may be more appropriate to detect abnormal movement patterns. However, the small difference in absolute values (i.e., degrees of movement) may indicate that the differences are not clinically significant, particularly for speed comparisons. Researchers and clinicians should take this into consideration when choosing the most appropriate task and the instruction to give during its execution.

Characterization of the complete mitochondrial genome of Neoasterolepisma foreli (Insecta: Zygentoma: Lepismatidae) and the phylogeny of basal Ectognatha.

The silverfish Neoasterolepisma foreli belongs to the family Lepismatidae within Zygentoma and is well known for the peculiar habit of living in strict association with ant nests (myrmecophily). In this study, we describe its mitochondrial genome, a circular molecule of 15,398 bp including the canonical 13 PCGs, 22 tRNAs, 2 rRNAs, as well as a 403 bp AT-rich region. A phylomitogenomic analysis of the new sequence, alongside basal hexapod mtDNAs, confirmed the monophyly of all orders, with some uncertainty over the position of the enigmatic Tricholepidion gertschi that would make Zygentoma paraphyletic. Neoasterolepisma foreli is recovered in a basal position within family Lepismatidae, at odd with our current understanding of the group that would, in turn, suggest a closer relationship with the genus Lepisma (Mendes, 1991).

The mitogenome of the true bug Nysius cymoides (Insecta, Heteroptera) and the phylogeny of Lygaeoidea.

The complete mitochondrial genome of the true bug (Homoptera) Nysius cymoides (Spinola, 1837) is herein described and used for phylogenetic comparison with other species of Lygaeoidea. The mtDNA has a gene order and other molecular features typically observed in hexapods, and a long A + T-rich region, due to the occurrence of several repeat units. The phylogenetic analyses support the monophyly of all families except Rhyparochromidae.

EZmito: a simple and fast tool for multiple mitogenome analyses.

Complete mitochondrial genome data are frequently applied to address phylogenetic/phylogeographic issues at different taxonomic levels in ecology and evolution. While sample preparation/sequencing is becoming more and more straightforward thanks to dropping costs for next-generation sequencing (NGS), data preparation and visualization remains a manually intensive step that may lead to errors if improperly conducted. We have elaborated, and here introduce, EZmito, a simple and intuitive, freely accessible Web Server aimed at automating some of these tasks. EZmito is divided into three main tools: EZpipe that assembles DNA matrices for phylo-mitogenomic analyses; EZskew that calculates genome, strand, and codon nucleotide compositional skews and EZcodon which computes Relative Synonymous Codon Usage statistics as well as amino acid usage frequency over multiple mitogenomes. Output is produced in tabular format as well as publication-quality graphics.

Fatigue Induced by Repeated Changes of Direction in Élite Female Football (Soccer) Players: Impact on Lower Limb Biomechanics and Implications for ACL Injury Prevention.

BACKGROUND: The etiology of Anterior Cruciate Ligament (ACL) injury in women football results from the interaction of several extrinsic and intrinsic risk factors. Extrinsic factors change dynamically, also due to fatigue. However, existing biomechanical findings concerning the impact of fatigue on the risk of ACL injuries remains inconsistent. We hypothesized that fatigue induced by acute workload in short and intense game periods, might in either of two ways: by pushing lower limbs mechanics toward a pattern close to injury mechanism, or alternatively by inducing opposed protective compensatory adjustments. AIM: In this study, we aimed at assessing the extent to which fatigue impact on joints kinematics and kinetics while performing repeated changes of direction (CoDs) in the light of the ACL risk factors. METHODS: This was an observational, cross-sectional associative study. Twenty female players (age: 20-31 years, 1st-2nd Italian division) performed a continuous shuttle run test (5-m) involving repeated 180°-CoDs until exhaustion. During the whole test, 3D kinematics and ground reaction forces were used to compute lower limb joints angles and internal moments. Measures of exercise internal load were: peak post-exercise blood lactate concentration, heart rate (HR) and perceived exertion. Continuous linear correlations between kinematics/kinetics waveforms (during the ground contact phase of the pivoting limb) and the number of consecutive CoD were computed during the exercise using a Statistical Parametric Mapping (SPM) approach. RESULTS: The test lasted 153 ± 72 s, with a rate of 14 ± 2 CoDs/min. Participants reached 95% of maximum HR and a peak lactate concentration of 11.2 ± 2.8 mmol/L. Exercise duration was inversely related to lactate concentration (r = -0.517, p < 0.01), while neither%HR max nor [La-] b nor RPE were correlated with test duration before exhaustion (p > 0.05). Alterations in lower limb kinematics were found in 100%, and in lower limb kinetics in 85% of the players. The most common kinematic pattern was a concurrent progressive reduction in hip and knee flexion angle at initial contact (10 players); 5 of them also showed a significantly more adducted hip. Knee extension moment decreased in 8, knee valgus moment increased in 5 players. A subset of participants showed a drift of pivoting limb kinematics that matches the known ACL injury mechanism; other players displayed less definite or even opposed behaviors. DISCUSSION: Players exhibited different strategies to cope with repeated CoDs, ranging from protective to potentially dangerous behaviors. While the latter was not a univocal effect, it reinforces the importance of individual biomechanical assessment when coping with fatigue.

Temporal variation in foraging activity and grouping patterns in a mountain-dwelling herbivore: Environmental and endogenous drivers.

In temperate ecosystems, seasonality influences animal behaviour. Food availability, weather, photoperiod and endogenous factors relevant to the biological cycle of individuals have been shown as major drivers of temporal changes in activity rhythms and group size/structure of herbivorous species. We evaluated how diurnal female foraging activity and grouping patterns of a mountain herbivore, the Apennine chamois Rupicapra pyrenaica ornata, varied during a decreasing gradient of pasture availability along the summer-autumn progression (July-October), a crucial period for the life cycle of mountain ungulates. Females increased diurnal foraging activity, possibly because of constrains elicited by variation in environmental factors. Size of mixed groups did not vary, in contrast with the hypothesis that groups should be smaller when pasture availability is lower. Proportion of females in groups increased, possibly suggesting that they concentrated on patchily distributed nutritious forbs. Occurrence of yearlings in groups decreased, which may have depended on dispersal of chamois in this age class. Presence of kids in groups did not show variation through summer-autumn, suggesting a close mother-juvenile relationship even at the end of weaning and/or, possibly, low summer mortality. Both endogenous and environmental factors contribute to shape variation in foraging activity and grouping behaviour in mountain-dwelling herbivores.