GWAS meta-analysis reveals key risk loci in essential tremor pathogenesis.

Essential tremor (ET) is a prevalent neurological disorder with a largely unknown underlying biology. In this genome-wide association study meta-analysis, comprising 16,480 ET cases and 1,936,173 controls from seven datasets, we identify 12 sequence variants at 11 loci. Evaluating mRNA expression, splicing, plasma protein levels, and coding effects, we highlight seven putative causal genes at these loci, including CA3 and CPLX1. CA3 encodes Carbonic Anhydrase III and carbonic anhydrase inhibitors have been shown to decrease tremors. CPLX1, encoding Complexin-1, regulates neurotransmitter release. Through gene-set enrichment analysis, we identify a significant association with specific cell types, including dopaminergic and GABAergic neurons, as well as biological processes like Rho GTPase signaling. Genetic correlation analyses reveals a positive association between ET and Parkinson's disease, depression, and anxiety-related phenotypes. This research uncovers risk loci, enhancing our knowledge of the complex genetics of this common but poorly understood disorder, and highlights CA3 and CPLX1 as potential therapeutic targets.

The structural repertoire of Fusarium oxysporum f. sp. lycopersici effectors revealed by experimental and computational studies.

Plant pathogens secrete proteins, known as effectors, that function in the apoplast or inside plant cells to promote virulence. Effector recognition by cell-surface or cytosolic receptors results in the activation of defence pathways and plant immunity. Despite their importance, our general understanding of fungal effector function and recognition by immunity receptors remains poor. One complication often associated with effectors is their high sequence diversity and lack of identifiable sequence motifs precluding prediction of structure or function. In recent years, several studies have demonstrated that fungal effectors can be grouped into structural classes, despite significant sequence variation and existence across taxonomic groups. Using protein X-ray crystallography, we identify a new structural class of effectors hidden within the secreted in xylem (SIX) effectors from Fusarium oxysporum f. sp. lycopersici (Fol). The recognised effectors Avr1 (SIX4) and Avr3 (SIX1) represent the founding members of the Fol dual-domain (FOLD) effector class, with members containing two distinct domains. Using AlphaFold2, we predicted the full SIX effector repertoire of Fol and show that SIX6 and SIX13 are also FOLD effectors, which we validated experimentally for SIX6. Based on structural prediction and comparisons, we show that FOLD effectors are present within three divisions of fungi and are expanded in pathogens and symbionts. Further structural comparisons demonstrate that Fol secretes effectors that adopt a limited number of structural folds during infection of tomato. This analysis also revealed a structural relationship between transcriptionally co-regulated effector pairs. We make use of the Avr1 structure to understand its recognition by the I receptor, which leads to disease resistance in tomato. This study represents an important advance in our understanding of Fol-tomato, and by extension plant-fungal interactions, which will assist in the development of novel control and engineering strategies to combat plant pathogens.

Effect of a Conical-PEP Mask on Exercise in Subjects With COPD.

BACKGROUND: Dynamic hyperinflation (DH) is a major pathophysiology of COPD that is directly related to dyspnea and exercise intolerance. Positive expiratory pressure (PEP) might reduce DH and dyspnea during exercise, but at present, there is insufficient evidence to conclude whether it is beneficial for DH, dyspnea, and exercise capacity in COPD. METHODS: A randomized crossover trial with concealed allocation was conducted in 37 moderate to very severe subjects with COPD (34 males, age 66.6 ± 7.4 y, FEV1% of predicted 56.3 ± 13.7). The experimental condition was conical-PEP breathing with a PEP of around 5 cm H2O during a spot marching exercise at a constant speed, inducing 71 ± 9% age-predicted maximum heart rate to symptom limit or 25 min. The control condition was usual breathing. Exercise endurance time and end-exercise symptoms were recorded. Inspiratory capacity (IC) was measured pre-exercise and immediately post exercise. Cardiopulmonary function and breathlessness were monitored throughout the test and after 10 min of recovery. RESULTS: There were no complications or adverse effects during exercise with a conical-PEP mask. Conical-PEP showed longer exercise times than control (median 11.0 [interquartile range 7.7-17.0] min vs 8 [6.0-11.5] min, respectively, P < .001). Most stopped exercising because of breathlessness and leg fatigue. At the end of exercise, IC and breathlessness showed non-significant differences between the conditions, but breathlessness was significantly lower in conical-PEP (median 4 [1.5-5.0] than control 5 [3-6] on Borg scale at isotime for control [8 min]). CONCLUSIONS: Breathing with a 5 cm H2O conical-PEP mask improved exercise time (median 27.1% [0.6-52.9]) in subjects with COPD. The improvement in exercise with the conical-PEP mask was associated with slower development of breathlessness, possibly due to delays in DH development.

Decrements of mobility and power in recreationally active septuagenarians is related to loss of force, but not slowing of the muscle: a 5-year longitudinal study.

A lesser 6-min walk distance (6MWD) and timed up-and-go (TUG) in old compared with young adults was previously linked to slowing of muscle contractile properties. The purpose of the present study was to determine whether any further reductions in 6MWD and TUG over a 5-year period in septuagenarians are associated with further slowing of muscle contractile properties. We measured muscle function by a countermovement jump, isometric maximal knee extensor strength (MVC) on a dynamometer and quadriceps muscle size by magnetic resonance imaging (MRI) in 17 older women (71.1 ± 2.8 y) and 17 older men (71.3 ± 4.1y). Performance in TUG and 6MWD were reduced over the 5-year period, irrespective of sex (P < 0.001), and both were correlated with power at both baseline and follow-up (R ≥ 0.53; P ≤ 0.001). Jump take-off velocity (VCMJ) was slower at follow-up (P < 0.01) and correlated with 6MWD and TUG at both baseline and follow-up in both sexes (R ≥ 0.54; P ≤ 0.001). However, the relationship between 'body mass: maximal muscle force ratio' with VCMJ was not significantly changed, indicating that the lower VCMJ was attributable to muscles working at a higher relative load, hence a lower part of the force-velocity relationship, due to a reduction in MVC (body mass had not changed significantly), rather than slowing of the muscle. The lower VCMJ in women than men (P < 0.001) was likewise attributable to a lower MVC rather than slower contractile properties in women. In conclusion, the decrement in 6MWD and TUG in septuagenarians is due to a loss of muscle mass, rather than further loss of muscle quality.

Genetic variants associated with syncope implicate neural and autonomic processes.

AIMS: Syncope is a common and clinically challenging condition. In this study, the genetics of syncope were investigated to seek knowledge about its pathophysiology and prognostic implications. METHODS AND RESULTS: This genome-wide association meta-analysis included 56 071 syncope cases and 890 790 controls from deCODE genetics (Iceland), UK Biobank (United Kingdom), and Copenhagen Hospital Biobank Cardiovascular Study/Danish Blood Donor Study (Denmark), with a follow-up assessment of variants in 22 412 cases and 286 003 controls from Intermountain (Utah, USA) and FinnGen (Finland). The study yielded 18 independent syncope variants, 17 of which were novel. One of the variants, p.Ser140Thr in PTPRN2, affected syncope only when maternally inherited. Another variant associated with a vasovagal reaction during blood donation and five others with heart rate and/or blood pressure regulation, with variable directions of effects. None of the 18 associations could be attributed to cardiovascular or other disorders. Annotation with regard to regulatory elements indicated that the syncope variants were preferentially located in neural-specific regulatory regions. Mendelian randomization analysis supported a causal effect of coronary artery disease on syncope. A polygenic score (PGS) for syncope captured genetic correlation with cardiovascular disorders, diabetes, depression, and shortened lifespan. However, a score based solely on the 18 syncope variants performed similarly to the PGS in detecting syncope risk but did not associate with other disorders. CONCLUSION: The results demonstrate that syncope has a distinct genetic architecture that implicates neural regulatory processes and a complex relationship with heart rate and blood pressure regulation. A shared genetic background with poor cardiovascular health was observed, supporting the importance of a thorough assessment of individuals presenting with syncope.

A rust-fungus Nudix hydrolase effector decaps mRNA in vitro and interferes with plant immune pathways.

To infect plants, pathogenic fungi secrete small proteins called effectors. Here, we describe the catalytic activity and potential virulence function of the Nudix hydrolase effector AvrM14 from the flax rust fungus (Melampsora lini). We completed extensive in vitro assays to characterise the enzymatic activity of the AvrM14 effector. Additionally, we used in planta transient expression of wild-type and catalytically dead AvrM14 versions followed by biochemical assays, phenotypic analysis and RNA sequencing to unravel how the catalytic activity of AvrM14 impacts plant immunity. AvrM14 is an extremely selective enzyme capable of removing the protective 5' cap from mRNA transcripts in vitro. Homodimerisation of AvrM14 promoted biologically relevant mRNA cap cleavage in vitro and this activity was conserved in related effectors from other Melampsora spp. In planta expression of wild-type AvrM14, but not the catalytically dead version, suppressed immune-related reactive oxygen species production, altered the abundance of some circadian-rhythm-associated mRNA transcripts and reduced the hypersensitive cell-death response triggered by the flax disease resistance protein M1. To date, the decapping of host mRNA as a virulence strategy has not been described beyond viruses. Our results indicate that some fungal pathogens produce Nudix hydrolase effectors with in vitro mRNA-decapping activity capable of interfering with plant immunity.

Transcriptome Analysis of Fusarium-Tomato Interaction Based on an Updated Genome Annotation of Fusarium oxysporum f. sp. lycopersici Identifies Novel Effector Candidates That Suppress or Induce Cell Death in Nicotiana benthamiana.

Fusarium oxysporum f. sp. lycopersici (Fol) causes vascular wilt disease in tomato. Upon colonization of the host, Fol secretes many small effector proteins into the xylem sap to facilitate infection. Besides known SIX (secreted in xylem) proteins, the identity of additional effectors that contribute to Fol pathogenicity remains largely unexplored. We performed a deep RNA-sequencing analysis of Fol race 2-infected tomato, used the sequence data to annotate a published genome assembly generated via PacBio SMRT sequencing of the Fol race 2 reference strain Fol4287, and analysed the resulting transcriptome to identify Fol effector candidates among the newly annotated genes. We examined the Fol-infection expression profiles of all 13 SIX genes present in Fol race 2 and identified 27 new candidate effector genes that were likewise significantly upregulated upon Fol infection. Using Agrobacterium-mediated transformation, we tested the ability of 22 of the new candidate effector genes to suppress or induce cell death in leaves of Nicotiana benthamiana. One effector candidate designated Fol-EC19, encoding a secreted guanyl-specific ribonuclease, was found to trigger cell death and two effector candidates designated Fol-EC14 and Fol-EC20, encoding a glucanase and a secreted trypsin, respectively, were identified that can suppress Bax-mediated cell death. Remarkably, Fol-EC14 and Fol-EC20 were also found to suppress I-2/Avr2- and I/Avr1-mediated cell death. Using the yeast secretion trap screening system, we showed that these three biologically-active effector candidates each contain a functional signal peptide for protein secretion. Our findings provide a basis for further understanding the virulence functions of Fol effectors.

Germline variants of ATG7 in familial cholangiocarcinoma alter autophagy and p62.

Autophagy is a housekeeping mechanism tasked with eliminating misfolded proteins and damaged organelles to maintain cellular homeostasis. Autophagy deficiency results in increased oxidative stress, DNA damage and chronic cellular injury. Among the core genes in the autophagy machinery, ATG7 is required for autophagy initiation and autophagosome formation. Based on the analysis of an extended pedigree of familial cholangiocarcinoma, we determined that all affected family members had a novel germline mutation (c.2000C>T p.Arg659* (p.R659*)) in ATG7. Somatic deletions of ATG7 were identified in the tumors of affected individuals. We applied linked-read sequencing to one tumor sample and demonstrated that the ATG7 somatic deletion and germline mutation were located on distinct alleles, resulting in two hits to ATG7. From a parallel population genetic study, we identified a germline polymorphism of ATG7 (c.1591C>G p.Asp522Glu (p.D522E)) associated with increased risk of cholangiocarcinoma. To characterize the impact of these germline ATG7 variants on autophagy activity, we developed an ATG7-null cell line derived from the human bile duct. The mutant p.R659* ATG7 protein lacked the ability to lipidate its LC3 substrate, leading to complete loss of autophagy and increased p62 levels. Our findings indicate that germline ATG7 variants have the potential to impact autophagy function with implications for cholangiocarcinoma development.

Investigating the relationship between euthanasia and/or assisted suicide and rates of non-assisted suicide: systematic review.

BACKGROUND: Euthanasia and assisted suicide (EAS) are practices that aim to alleviate the suffering of people with life-limiting illnesses, but are controversial. One area of debate is the relationship between EAS and suicide rates in the population, where there have been claims that availability of EAS will reduce the number of self-initiated deaths (EAS and suicide combined). Others claim that legislation for EAS makes it acceptable to end one's own life, a message at variance with that of suicide prevention campaigns. AIMS: To examine the relationship between the introduction of EAS and rates of non-assisted suicide and self-initiated death. METHOD: We conducted a systematic review to examine the association between EAS and rates of non-assisted suicide and of self-initiated death. We searched PubMed, Scopus, PsycINFO and Science Direct, until 20 December 2021. Studies that examined EAS and reported data on population-based suicide rates were included. RESULTS: Six studies met the inclusion criteria; four reported increases in overall rates of self-initiated death and, in some cases, increased non-assisted suicide. This increase in non-assisted suicide was mostly non-significant when sociodemographic factors were controlled for. Studies from Switzerland and Oregon reported elevated rates of self-initiated death among older women, consistent with higher rates of depressive illnesses in this population. CONCLUSIONS: The findings of this review do not support the hypothesis that introducing EAS reduces rates of non-assisted suicide. The disproportionate impact on older women indicates unmet suicide prevention needs in this population.

Optimized Production of Disulfide-Bonded Fungal Effectors in Escherichia coli Using CyDisCo and FunCyDisCo Coexpression Approaches.

Effectors are a key part of the arsenal of plant-pathogenic fungi and promote pathogen virulence and disease. Effectors typically lack sequence similarity to proteins with known functional domains and motifs, limiting our ability to predict their functions and understand how they are recognized by plant hosts. As a result, cross-disciplinary approaches involving structural biology and protein biochemistry are often required to decipher and better characterize effector function. These approaches are reliant on high yields of relatively pure protein, which often requires protein production using a heterologous expression system. For some effectors, establishing an efficient production system can be difficult, particularly those that require multiple disulfide bonds to achieve their naturally folded structure. Here, we describe the use of a coexpression system within the heterologous host Escherichia coli, termed CyDisCo (cytoplasmic disulfide bond formation in E. coli) to produce disulfide bonded fungal effectors. We demonstrate that CyDisCo and a naturalized coexpression approach termed FunCyDisCo (Fungi CyDisCo) can significantly improve the production yields of numerous disulfide-bonded effectors from diverse fungal pathogens. The ability to produce large quantities of functional recombinant protein has facilitated functional studies and crystallization of several of these reported fungal effectors. We suggest this approach could be broadly useful in the investigation of the function and recognition of a broad range of disulfide bond-containing effectors.[Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Informing radiotherapy decisions in stage I/IIa Hodgkin lymphoma: modeling life expectancy using radiation dosimetry.

In recent randomized trials, omitting consolidative radiotherapy (RT) in early-stage Hodgkin lymphoma (ESHL) increased relapses. However, decades of follow-up are required to observe whether lower initial disease control is compensated by reduced risk of late effects. Extrapolation beyond trial follow-up is therefore necessary to inform current treatment decisions. To this end, we developed a microsimulation model to estimate lifetime quality-adjusted life years (QALYs) after combined modality treatment (CMT) or chemotherapy-alone for stage I/IIa ESHL. For CMT, the model included risks of breast and lung cancer, coronary heart disease, and ischemic stroke. Comparative outcomes were assessed for a clinically relevant range of example patients differing by age, sex, smoking status, and representative organs at risk (OAR) radiation doses informed by the RAPID trial. Analysis was performed with and without a 3.5% discount rate on future health. Smoking status had a large effect on optimal treatment choice. CMT was superior for nearly all never smoker example patients regardless of age, sex, and OAR doses. At a maximum, CMT produced a 1.095 (95% CI: 1.054-1.137) gain in undiscounted QALYs for a 20-year-old male never smoker with unilateral neck disease. In contrast, current smokers could substantially gain from chemotherapy-alone treatment. Again at a maximum, a 20-year-old male current smoker with bilateral neck and whole mediastinum involvement gained 3.500 (95% CI: 3.400 to 3.600) undiscounted QALYs with chemotherapy-alone treatment. Overall, CMT was more favorable the younger the patient, when future health discounting was included, and in never smokers.

A genome-wide meta-analysis uncovers six sequence variants conferring risk of vertigo.

Vertigo is the leading symptom of vestibular disorders and a major risk factor for falls. In a genome-wide association study of vertigo (Ncases = 48,072, Ncontrols = 894,541), we uncovered an association with six common sequence variants in individuals of European ancestry, including missense variants in ZNF91, OTOG, OTOGL, and TECTA, and a cis-eQTL for ARMC9. The association of variants in ZNF91, OTOGL, and OTOP1 was driven by an association with benign paroxysmal positional vertigo. Using previous reports of sequence variants associating with age-related hearing impairment and motion sickness, we found eight additional variants that associate with vertigo. Although disorders of the auditory and the vestibular system may co-occur, none of the six genome-wide significant vertigo variants were associated with hearing loss and only one was associated with age-related hearing impairment. Our results uncovered sequence variants associating with vertigo in a genome-wide association study and implicated genes with known roles in inner ear development, maintenance, and disease.

Did the Pope ban blood transfusion in 1678?

This paper concerns an alleged event in the history of haematology that disrupts the otherwise positive narrative of papal encouragement for blood transfusion. It is frequently stated, in popular websites and in the scholarly literature, that when blood transfusion was first developed it was banned by the Pope. However, careful analysis of the sources cited shows this claim to be without historical foundation. There was never any papal prohibition of blood transfusion. It is a myth that needs to be dispelled if the full extent of the Catholic Church's support for blood and organ donation is to be appreciated.

The crystal structure of SnTox3 from the necrotrophic fungus Parastagonospora nodorum reveals a unique effector fold and provides insight into Snn3 recognition and pro-domain protease processing of fungal effectors.

Plant pathogens cause disease through secreted effector proteins, which act to promote infection. Typically, the sequences of effectors provide little functional information and further targeted experimentation is required. Here, we utilized a structure/function approach to study SnTox3, an effector from the necrotrophic fungal pathogen Parastagonospora nodorum, which causes cell death in wheat-lines carrying the sensitivity gene Snn3. We developed a workflow for the production of SnTox3 in a heterologous host that enabled crystal structure determination and functional studies. We show this approach can be successfully applied to study effectors from other pathogenic fungi. The ß-barrel fold of SnTox3 is a novel fold among fungal effectors. Structure-guided mutagenesis enabled the identification of residues required for Snn3 recognition. SnTox3 is a pre-pro-protein, and the pro-domain of SnTox3 can be cleaved in vitro by the protease Kex2. Complementing this, an in silico study uncovered the prevalence of a conserved motif (LxxR) in an expanded set of putative pro-domain-containing fungal effectors, some of which can be cleaved by Kex2 in vitro. Our in vitro and in silico study suggests that Kex2-processed pro-domain (designated here as K2PP) effectors are common in fungi and this may have broad implications for the approaches used to study their functions.

Response of Knee Extensor Muscle-Tendon Unit Stiffness to Unaccustomed and Repeated High-Volume Eccentric Exercise.

The purposes of this study were to investigate the muscle-tendon unit stiffness response and to compare the stiffness with those of other indirect markers induced by two bouts of unaccustomed eccentric exercise. Eleven untrained men performed two bouts of 200 maximal eccentric contractions of the right quadriceps 4 weeks apart. Changes in stiffness, pain evoked by stretching and pressure, plasma creatine kinase (CK) activity, and muscle thickness were followed for 7 days after each bout. Stiffness and pain peaked immediately and 1 day after the first exercise bout, whereas CK and thickness were highest 4 and 7 days after the first exercise bout, respectively (p < 0.05 for all). Muscular pain, thickness, and stiffness responses were lower by 53.3%, 99%, and 11.6%, respectively, after the repeated bout compared to after the first bout (p < 0.05 for all), while CK activity response did not differ significantly between bouts. High responders for an increase in muscle-tendon unit stiffness showed a repeated-bout effect for stiffness, pain, and CK activity (by 29%, 65%, and 98%, p < 0.05 for all), but the repeated-bout effect was not that clear in low responders. These findings suggest that a repeated eccentric exercise bout effect on stiffness in quadriceps is mostly not associated with muscle pain and CK activity, but there are large individual differences.

Stair Gait in Older Adults Worsens With Smaller Step Treads and When Transitioning Between Level and Stair Walking.

Older people have an increased risk of falling during locomotion, with falls on stairs being particularly common and dangerous. Step going (i.e., the horizontal distance between two consecutive step edges) defines the base of support available for foot placement on stairs, as with smaller going, the user's ability to balance on the steps may become problematic. Here we quantified how stair negotiation in older participants changes between four goings (175, 225, 275, and 325 mm) and compared stair negotiation with and without a walking approach. Twenty-one younger (29 ± 6 years) and 20 older (74 ± 4 years) participants negotiated a 7-step experimental stair. Motion capture and step-embedded force platform data were collected. Handrail use was also monitored. From the motion capture data, body velocity, trunk orientation, foot clearance and foot overhang were quantified. For all participants, as stair going decreased, gait velocity (ascent pA = 0.033, descent pD = 0.003) and horizontal step clearance decreased (pA = 0.001), while trunk rotation (pD = 0.002) and foot overhang increased (pA,D < 0.001). Compared to the younger group, older participants used the handrail more, were slower across all conditions (pA < 0.001, pD = 0.001) and their foot clearance tended to be smaller. With a walking approach, the older group (Group x Start interaction) showed a larger trunk rotation (pA = 0.011, pD = 0.015), and smaller lead foot horizontal (pA = 0.046) and vertical clearances (pD = 0.039) compared to the younger group. A regression analysis to determine the predictors of foot clearance and amount of overhang showed that physical activity was a common predictor for both age groups. In addition, for the older group, medications and fear of falling were found to predict stair performance for most goings, while sway during single-legged standing was the most common predictor for the younger group. Older participants adapted to smaller goings by using the handrails and reducing gait velocity. The predictors of performance suggest that motor and fall risk assessment is complex and multifactorial. The results shown here are consistent with the recommendation that larger going and pausing before negotiating stairs may improve stair safety, especially for older users.

The Aspergillus nidulans IQGAP orthologue SepG is required for constriction of the contractile actomyosin ring.

In this research we report that the sepG1 mutation in Aspergillus nidulans resides in gene AN9463, which is predicted to encode an IQGAP orthologue. The genetic lesion is predicted to result in a G-to-R substitution at residue 1637 of the 1737-residue protein in a highly conserved region of the RasGAP-C-terminal (RGCT) domain. When grown at restrictive temperature, strains expressing the sepGG1637R (sepG1) allele are aseptate, with reduced colony growth and aberrantly formed conidiophores. The aseptate condition can be replicated by deletion of AN9463 or by downregulating its expression via introduced promoters. The mutation does not prevent assembly of a cortical contractile actomyosin ring (CAR) at putative septation sites, but tight compaction of the rings is impaired and the rings fail to constrict. Both GFP::SepG wild type and the GFP-tagged product of the sepG1 allele localize to the CAR at both permissive and restrictive temperatures. Downregulation of myoB (encoding the A. nidulans type-II myosin heavy chain) does not prevent formation of SepG rings at septation sites, but filamentous actin is required for CAR localization of SepG and MyoB. We identify fourteen probable IQ-motifs (EF-hand protein binding sites) in the predicted SepG sequence. Two of the A. nidulans EF-hand proteins, myosin essential light chain (AnCdc4) and myosin regulatory light chain (MrlC), colocalize with SepG and MyoB at all stages of CAR formation and constriction. However, calmodulin (CamA) appears at septation sites only after the CAR has become fully compacted. When expression of sepG is downregulated, leaving MyoB as the sole IQ-motif protein in the pre-compaction CAR, both MrlC and AnCdc4 continue to associate with the forming CAR. When myoB expression is downregulated, leaving SepG as the sole IQ-motif protein in the CAR, AnCdc4 association with the forming CAR continues but MrlC fails to associate. This supports a model in which the IQ motifs of MyoB bind both MrlC and AnCdc4, while the IQ motifs of SepG bind only AnCdc4. Downregulation of either mrlC or Ancdc4 results in an aseptate phenotype, but has no effect on association of either SepG or MyoB with the CAR.

Are Force Enhancement after Stretch and Muscle Fatigue Due to Effects of Elevated Inorganic Phosphate and Low Calcium on Cross Bridge Kinetics?

Background and Objectives: Muscle fatigue is characterised by (1) loss of force, (2) decreased maximal shortening velocity and (3) a greater resistance to stretch that could be due to reduced intracellular Ca2+ and increased Pi, which alter cross bridge kinetics. Materials and Methods: To investigate this, we used (1) 2,3-butanedione monoxime (BDM), believed to increase the proportion of attached but non-force-generating cross bridges; (2) Pi that increases the proportion of attached cross bridges, but with Pi still attached; and (3) reduced activating Ca2+. We used permeabilised rat soleus fibres, activated with pCa 4.5 at 15 °C. Results: The addition of 1 mM BDM or 15 mM Pi, or the lowering of the Ca2+ to pCa 5.5, all reduced the isometric force by around 50%. Stiffness decreased in proportion to isometric force when the fibres were activated at pCa 5.5, but was well maintained in the presence of Pi and BDM. Force enhancement after a stretch increased with the length of stretch and Pi, suggesting a role for titin. Maximum shortening velocity was reduced by about 50% in the presence of BDM and pCa 5.5, but was slightly increased by Pi. Neither decreasing Ca2+ nor increasing Pi alone mimicked the effects of fatigue on muscle contractile characteristics entirely. Only BDM elicited a decrease of force and slowing with maintained stiffness, similar to the situation in fatigued muscle. Conclusions: This suggests that in fatigue, there is an accumulation of attached but low-force cross bridges that cannot be the result of the combined action of reduced Ca2+ or increased Pi alone, but is probably due to a combination of factors that change during fatigue.

Low frequency fatigue and changes in muscle fascicle length following eccentric exercise of the knee extensors.

NEW FINDING: What is the central question of this study? Does low frequency muscle fatigue indicate a failure of excitation-contraction coupling after eccentric exercise, or is it simply due to a change in muscle length? What is the main finding and its importance? The low to high frequency muscle fatigue ratio was relatively insensitive to changes in muscle length, and any changes in length following eccentric exercise were far too small to account for the high degree of low frequency fatigue. The results strengthen the suggestion that the early loss of force following eccentric exercise is due to a deficit of excitation-contraction coupling. ABSTRACT: Development of long lasting fatigue (low frequency fatigue; LFF), assessed as the ratio of forces at 20 and 100 Hz stimulation, suggests the early phase of muscle damage caused by eccentric exercise is due to a deficit of excitation-contraction coupling. However, this could be caused by a change of muscle length. Eleven men (21.3 ± 2.0 years) performed 200 maximum eccentric knee extensions (30-110 deg flexion). Force generated by 20 and 100 Hz stimulation and maximum isometric force (MIF) were determined at knee angles 50, 70 and 90 deg before and immediately after the exercise. Vastus lateralis fascicle length (FL) was measured by ultrasound of resting and contracting muscle. Peak MIF (829 ± 119 N) was at 70 deg knee flexion, falling to 486 ± 180 N (P < 0.001) after exercise, but with no change in optimum angle. FLs at rest were unaffected by eccentric exercise, but during contraction they were on average 8.8% (95% CI: 4.1, 13.5%, P = 0.002) longer after exercise. Before exercise, the 20/100 ratio increased with muscle length, from 0.69 ± 0.09 at 50 deg, 0.72 ± 0.05 at 70 deg and 0.80 ± 0.08 at knee angle 90 deg (P < 0.001). After eccentric exercise the 20/100 ratio was reduced to 0.29 ± 0.08 at 50 deg, 0.27 ± 0.04 at 70 deg and 0.34 ± 0.04 at 90 deg (P < 0.001). The 20/100 ratio was relatively insensitive to changes in muscle length and the decrease following eccentric exercise was far greater than might be caused by any changes in muscle length after eccentric exercise. The results show that LFF following eccentric exercise is not due to change in muscle length and strengthen the suggestion that it represents a deficit in excitation-contraction coupling.

Five-year longitudinal changes in thigh muscle mass of septuagenarian men and women assessed with DXA and MRI.

Magnetic resonance imaging (MRI) and dual-energy X-ray absorptiometry (DXA) were used to assess changes in thigh lean mass in septuagenarian men and women during a 5-year longitudinal study. Twenty-four older individuals participated in the study (10 men: 71.6 ± 4.1 years; 14 women: 71.3 ± 3.2 years at baseline). Thigh MRI and whole-body DXA scans were used to estimate changes in thigh lean mass. Both MRI and DXA showed that thigh lean mass was reduced by approximately 5% (P = 0.001) over the 5-year period in both men and women. The percentage loss of muscle mass determined with MRI and DXA showed moderate correlation (R2 = 0.466; P < 0.001). Bland-Altman analysis showed that the average change over 5 years of follow-up measured by DXA was only 0.18% greater than MRI, where the limits of agreement between DXA and MRI were ± 10.4%. Baseline thigh lean mass did not predict the percentage loss of thigh lean mass over the 5-year period (R2 = 0.003; P = 0.397), but a higher baseline body fat percentage was associated with a larger loss of thigh muscle mass in men (R2 = 0.677; P < 0.003) but not in women (R2 = 0.073; P < 0.176). In conclusion, (1) DXA and MRI showed a similar percentage loss of muscle mass over a 5-year period in septuagenarian men and women that (2) was independent of baseline muscle mass, but (3) increased with higher baseline body fat percentage in men.