Biology subjects

Carbonaro, M.

Publications and source records attributed to Carbonaro, M..

4 recordsLinked to original sources

bioRxiv2025

Motor unit behaviour adaptations across the lifespan: sex differences in young, middle-aged and old adults

Ageing is associated with neuromuscular decline, and emerging evidence suggests that sex may influence the time course of motor unit adaptations. This study examined age- and sex-related differences in motor unit firing behaviour across young (YG), middle-aged (MA), and older adults (OLD), by integrating high-density EMG motor unit analysis with muscle morphology and daily physical activity measurements. The analysis of single motor unit activity during submaximal isometric contractions of the vastus lateralis revealed that older adults had lower firing rates and a reduced capacity to modulate discharge frequency during force-increasing contractions. In the YG and MA groups, females showed higher motor unit firing rates and variability than males, while in OLD these sex differences were no longer present. Females also demonstrated a steeper decline in firing rate modulation between MA and OLD. Reductions in muscle cross-sectional area and thickness were similar between sexes. Physical activity levels declined with age in both sexes. These findings reveal distinct, sex-specific trajectories of neuromuscular ageing, with females showing greater motor neuron function decline between MA and OLD, in the absence of sex-related differences in the rate of morphological deterioration. The attenuation of sex differences in older age suggests a convergence of neuromuscular profiles with ageing. While physical activity may contribute to the observed sex-specific patterns, other mechanisms related to hormonal shifts warrant further investigation. These insights underscore the importance of considering age and sex in the study of motor control and in the development of targeted interventions to preserve muscle function across the lifespan.

neuroscience↗

bioRxiv2023

Combining high-density electromyography and ultrafast ultrasound to assess individual motor unit properties in vivo

This study aims to compare two methods for the identification of anatomical and mechanical motor unit (MU) properties through the integration of high-density surface electromyography (HDsEMG) and ultrafast ultrasound (UUS). The two approaches rely on a combined analysis of the firing pattern of active MUs, identified from HDsEMG, and tissue velocity sequences of the muscle cross-section, obtained from UUS. The first method is the spike-triggered averaging (STA) of the tissue velocity sequence based on the occurrences of MU firings. The second is a method based on spatio-temporal independent component analysis (STICA) enhanced with the information of single MU firings. We compared the capability of these two approaches to identify the regions where single MU fibers are located within the muscle cross-section (MU displacement area) in vivo. HDsEMG signals and UUS images were detected simultaneously from biceps brachii in ten participants (6 males and 4 females) during low-level isometric elbow flexions. Experimental signals were processed by implementing both STA and STICA approaches. The medio-lateral distance between the estimated MU displacement areas and the centroid of the MU action potential distributions was used to compare the two methods. We found that STICA and STA are able to detect MU displacement areas. However, STICA provides more precise estimations to the detriment of higher computational complexity.

bioengineering↗

bioRxiv2023

Spatially repeatable components from ultrafast ultrasound are associated with motor unit activity in human isometric contractions

ObjectiveUltrafast ultrasound imaging has been used to measure intramuscular mechanical dynamics associated with single motor unit (MU) activations. Detecting MU activity from ultrasound sequences requires decomposing a displacement velocity field into components consisting of spatial maps and temporal displacement signals. These components can be associated with putative MU activity or spurious movements (noise). The differentiation between putative MUs and noise has been accomplished by comparing the temporal displacement signals with MU firings obtained from needle EMG. Here, we examined whether the repeatability of the spatial maps over brief time intervals can serve as a criterion for distinguishing putative MUs from noise in low-force isometric contractions. ApproachIn five healthy subjects, ultrafast ultrasound images and high-density surface EMG (HDsEMG) were recorded simultaneously from biceps brachii. MUs identified through HDsEMG decomposition were used as a reference to assess the outcomes of the ultrasound-based decomposition. For each contraction, displacement velocity sequences from the same eight-second ultrasound recording were separated into consecutive two-second epochs and decomposed. The Jaccard Similarity Coefficient (JSC) was employed to evaluate the repeatability of components spatial maps across epochs. Finally, the association between the ultrasound components and the MUs decomposed from HDsEMG was assessed. Main resultsAll the MU-matched components had JSC > 0.38, indicating they were repeatable and accounted for about one-third of the HDsEMG-detected MUs (1.8 {+/-} 1.6 matches over 4.9 {+/-} 1.8 MUs). The repeatable components (with JSC over the empirical threshold of 0.38) represented 14% of the total components (6.5 {+/-} 3.3 components). These findings align with our hypothesis that intra-sequence repeatability can differentiate putative MUs from spurious components and can be used for data reduction. SignificanceThe results of our study provide the foundation for developing stand-alone methods to identify MU in ultrafast ultrasound sequences and represent a step forward towards real-time imaging of active MU territories. These methods are relevant for studying muscle neuromechanics and designing novel neural interfaces.

bioengineering↗

bioRxiv2021

A hyperthermoactive-Cas9 editing tool reveals the role of a unique arsenite methyltransferase in the arsenic resistance system of Thermus thermophilus HB27

Arsenic detoxification systems can be found in a wide range of organisms, from bacteria to man. In a previous study, we discovered an arsenic-responsive transcriptional regulator in the thermophilic bacterium Thermus thermophilus HB27 (TtSmtB). Here, we characterize the arsenic resistance system of T. thermophilus in more detail. We employed TtSmtB-based pull-down assays with protein extracts from cultures treated with arsenate and arsenite to obtain an S-adenosyl-Lmethionine (SAM)-dependent arsenite methyltransferase (TtArsM). In vivo and in vitro analyses were performed to shed light on this new component of the arsenic resistance network and its peculiar catalytic mechanism. Heterologous expression of TtarsM in Escherichia coli resulted in arsenite detoxification at mesophilic temperatures. Although TtArsM does not contain a canonical arsenite binding site, the purified protein does catalyze SAM- dependent arsenite methylation with formation of monomethylarsenite (MMAs) and dimethylarsenite (DMAs). In addition, in vitro analyses confirmed the unique interaction between TtArsM and TtSmtB. Next, a highly efficient ThermoCas9- based genome-editing tool was developed to delete the TtArsM-encoding gene on the T. thermophilus genome and to confirm its involvement in the arsenite detoxification system. Finally, the TtarsX efflux pump gene in the T. thermophilus {Delta}TtarsM genome was substituted by a gene encoding a stabilized yellow fluorescent protein (sYFP) to create a sensitive genome-based bioreporter system for the detection of arsenic ions. ImportanceWe here describe the discovery of an unknown protein by using a proteomic approach with a functionally related protein as bait. Remarkably, we successfully obtained a novel type of enzyme through the interaction with a transcription regulator, controlling the expression of this enzyme. Employing this strategy, we isolated TtArsM, the first thermophilic prokaryotic arsenite methyltransferase, as a new enzyme of the arsenic resistance mechanism in T. thermophilus HB27. The atypical arsenite binding site of TtArsM categorizes the enzyme as the first member of a new arsenite methyltransferase type, exclusively present in the Thermus genus. The enzyme methylates arsenite producing MMAs and DMAs. Furthermore, we developed an hyperthermophilic Cas9-based genome-editing tool, active up to 65{degrees}C. The tool allowed us to perform highly efficient, marker-free modifications (either gene deletion or insertion) in the T. thermophilus genome. With these modifications, we confirmed the critical role of TtArsM in the arsenite detoxification system and developed a sensitive whole cell bioreporter for arsenic ions. We anticipate that the developed tool can be easily adapted for editing the genomes of other thermophilic bacteria, significantly boosting fundamental and metabolic engineering in hyperthermophilic microorganisms.

biochemistry↗