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Cellular processes require precise and specific gene regulation, in which continuous mRNA degradation is a major element. The mRNA degradation mechanisms should be able to degrade a wide range of different RNA substrates with high efficiency, but should at the same time be limited, to avoid killing the cell by elimination of all cellular RNA. RNase Y is a major endoribonuclease found in most Firmicutes, including Bacillus subtilis and Staphylococcus aureus. However, the molecular interactions that direct RNase Y to cleave the correct RNA molecules at the correct position remain unknown. In this work we have identified transcripts that are homologs in S. aureus and B. subtilis, and are RNase Y targets in both bacteria. Two such transcript pairs were used as models to show a functional overlap between the S. aureus and the B. subtilis RNase Y, which highlighted the importance of the nucleotide sequence of the RNA molecule itself in the RNase Y targeting process. Cleavage efficiency is driven by the primary nucleotide sequence immediately downstream of the cleavage site and base-pairing in a secondary structure a few nucleotides downstream. Cleavage positioning is roughly localised by the downstream secondary structure and fine-tuned by the nucleotide immediately upstream of the cleavage. The identified elements were sufficient for RNase Y-dependent cleavage, since the sequence elements from one of the model transcripts were able to convert an exogenous non-target transcript into a target for RNase Y.
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Bacillus subtilis , Regulación Bacteriana de la Expresión Génica , División del ARN , Estabilidad del ARN , ARN Bacteriano , Staphylococcus aureus , Staphylococcus aureus/genética , Staphylococcus aureus/enzimología , Bacillus subtilis/genética , Bacillus subtilis/enzimología , Bacillus subtilis/metabolismo , ARN Bacteriano/metabolismo , ARN Bacteriano/genética , Estabilidad del ARN/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Endorribonucleasas/metabolismo , Endorribonucleasas/genética , Conformación de Ácido Nucleico , Secuencia de BasesRESUMEN
The literature offers limited information on the effect of obesity on the rate of force development (RFD), a critical parameter for mobility in older adults. The objectives of this study were to explore the influence of obesity on the RFD in older adults and to examine the association between this neuromuscular parameter and walking speed. The participants (42 older adults) were classified into two groups: the control group (CG, n = 22; mean age = 81.13 ± 4.02 years; body mass index (BMI) = 25.13 ± 3.35 kg/m2), and the obese group (OG, n = 20; mean age = 77.71 ± 2.95 years; BMI = 34.46 ± 3.25 kg/m2). Walking speed (m/s) was measured using the 10 m walking test. Neuromuscular parameters of the plantar flexors were evaluated during a maximal voluntary contraction test using a dynamometer. The RFD was calculated from the linear slop of the force-time curve in the following two phases: from the onset of the contraction to 50 ms (RFD0-50) and from 100 to 200 ms (RFD100-200). The gait speed was lower in the OG compared to the CG (p < 0.001). The RFD50/100 and RFD100/200 were lower in the OG compared to the CG (p < 0.001). The RFD50/100 was found to be the predominant influencer on gait speed in the OG. In conclusion, obesity negatively impacts the RFD in older adults and RFD stands out as the primary factor among the studied parameters influencing gait speed.
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BACKGROUND: Excessive body weight is associated with gait alterations. In none of previous studies, body fat distribution has been considered as a factor that could change gait parameters and induce different neuromuscular adaptations. OBJECTIVE: This multicenter, analytical, and cross-sectional study aimed to investigate the influence of the body mass distribution on gait parameters and ankle muscle coactivation in obese individuals. METHODS: Three distinct groups were included in the study: a non-obese control group (CG, n = 15, average age = 32.8 ± 6.5 years, BMI = 21.4 ± 2.2 kg/m2), an obese-android group characterized by a Waist to Hip Ratio (WHR) greater than 1 (OAG, n = 15, age = 32.4 ± 3.9 years, BMI = 41.4 ± 3.9 kg/m2, WHR = 1.2 ± 0.2), and an obese-gynoid group with a WHR less than 1 (OGG, n = 15, age = 35.4 ± 4.1 years, BMI = 40.0 ± 5.7 kg/m2, WHR = 0.82 ± 0.3). All participants walked on an instrumented gait analysis treadmill at their self-selected walking speed for one minute. Spatiotemporal parameters, walking cycle phases, vertical ground reaction force (GRFv) and center of pressure (CoP) velocity were sampled from the treadmill software. Electromyography (EMG) activity of the gastrocnemius medialis (GM), the soleus (SOL) and tibialis anterior (TA) were collected during walking and used to calculate coactivation indexes (CI) between ankle plantar and dorsal flexors (GM/TA and SOL/TA) for the different walking cycle phases. RESULTS: Compared to OAG, OGG walked with shorter and larger strides, lower CoP velocity and GRFv. During the single support phase, SOL/TA coactivation was higher in OAG compared to OGG (p < .05). During the propulsion phase, SOL/TA coactivation was higher in OGG compared to OAG (p < .05). CONCLUSION: Gait parameters and ankle muscle coactivation in obese individuals seem to be strongly dependent on body mass distribution. From the biomechanical point of view, body mass distribution changes gait strategies in obese individuals inducing different neuromuscular adaptations during the single support and propulsion phases.
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Tobillo , Marcha , Humanos , Adulto , Estudios Transversales , Marcha/fisiología , Caminata/fisiología , Músculo Esquelético/fisiología , Electromiografía , Obesidad , Fenómenos BiomecánicosRESUMEN
Sarcopenic obesity (SO), characterized by age-related muscle loss and excess body fat, significantly impairs postural control. However, limited research has explored the effects of concurrent exercise training on neuromuscular strategies during postural control in older adults with SO. The study enrolled 50 older adults with SO, split into an intervention group (IG, n = 25, mean age = 76.1 ± 3.5 years; mean BMI = 34.4 ± 4.0 kg/m2) and a control group (CG, n = 25, mean age = 75.9 ± 5.4 years; mean BMI = 32.9 ± 2.3 kg/m2). Participants in the IG were engaged in 60-min Total Mobility Plus Program (TMP) sessions three times a week for four months, while the CG maintained their typical daily activities. Standardized evaluations were conducted both before and after the intervention. These assessments included the Romberg and Timed Up and Go (TUG) tests, as well as the measurement of Center of Pressure (CoP) displacements parameters under various conditions. Additionally, ankle muscle activities were quantified during postural control evaluations and maximal voluntary contractions of plantar and dorsal flexors. Post-intervention results revealed a significant reduction of the standing time measured in the Romberg (-15.6%, p < 0.005) and TUG (-34.6%, p < 0.05) tests. Additionally, CoP area and velocity were notably reduced in various conditions (p < 0.05). Postural control improvements were associated with an increase of strength (p < 0.05) and decrease of ankle muscle activation (p < 0.05). These findings highlight the reversibility of neuromuscular system alterations associated with the synergistic effects of sarcopenia and obesity, emphasizing the trainability of postural control regulation within this population. By incorporating these insights into clinical practice and public health strategies, it seems possible to optimize the health and well-being of older adults with SO.
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An EPR spectrum or an EPR sinogram for imaging contains information about all the paramagnetic species that are in the analyzed sample. When only one species is present, an image of its spatial repartition can be reconstructed from the sinogram by using the well-known Filtered Back-Projection (FBP). However, in the case of several species, the FBP does not allow the reconstruction of the images of each species from a standard acquisition. One has to use for this spectral-spatial imaging whose acquisition can be very long. A new approach, based on Total Variation minimization, is proposed in order to efficiently extract the spatial repartitions of all the species present in a sample from standard imaging data and therefore drastically reduce the acquisition time. Experiments have been carried out on Tetrathiatriarylmethyl, nitroxide and DPPH.
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Purpose: Cognitive impairment secondary to traumatic brain injury (TBI) is difficult to treat and usually results in severe disability. Method: A 48-year-old man presented with chronic refractory headaches and persistent disabling cognitive impairment after TBI. He was first treated with occipital nerve stimulation (ONS) implanted bilaterally to relieve headaches (8 years after the head trauma). Two years later, he was treated with a 6-week protocol combining repetitive transcranial magnetic stimulation (rTMS) delivered to multiple cortical sites (prefrontal cortex, language areas, and areas involved in visuo-spatial functions) and computerized cognitive training (CogT) (targeting memory, language, and visuo-spatial functions) to improve cognitive performance. Results: Executive and cognitive functions (attention, ability to perform calculations, and verbal fluency) improved in association with pain relief after ONS (33-42% improvement) and then improved even more after the rTMS-CogT protocol with an additional improvement of 36-40% on apathy, depression, and anxiety, leading to a significant reduction in caregiver burden. The functional improvement persisted and even increased at 6 months after the end of the rTMS-CogT procedure (10 years after the onset of TBI and 2 years after ONS implantation). Conclusion: This is the first observation describing sustained improvement in post-TBI refractory headache, depression, and cognitive impairment by the association of bilaterally implanted ONS and a combined procedure of multisite rTMS and CogT to target various brain functions.
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The potential impact of a specific physical activity program on biomechanical gait parameters and neuromuscular strategies around the ankle joint in older adults with sarcopenic obesity (SO) remains largely unexplored. The objective of this study was to investigate the effectiveness of a 24-week posture, strengthening, and motricity (PSM) program on improving neuromuscular strategies and biomechanical gait parameters in older adults with SO. 40 participants were randomly assigned to either the trained group (TG) and the control group (CG). Only the TG received the PSM program. Standardized evaluations were performed before and after the intervention, including walking tests on an instrumented gait analysis treadmill to evaluate biomechanical gait parameters and EMG activity of ankle muscles. After the PSM program, TG exhibited an increase in comfortable walking speed (+80%, p < 0.001) and step length (+38%, p < 0.05). Moreover, TG demonstrated a reduction in CoP velocity (-26%, p < 0.01). These gait modifications were associated with decreased muscle activity during the different gait phases (p < 0.05). The PSM program effectively improved gait and neuromuscular capacities in older adults with SO. Notably, these results shed light on the remarkable trainability of neuromuscular capacities in older adults with SO, despite the adverse effects of aging and obesity.
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BACKGROUND: Chronic kidney disease (CKD) induces muscle wasting and a reduction in the maximum voluntary force (MVF). Little is known about the neuromuscular fatigability in CKD patients, defined as the reduction of muscle force capacities during exercise. Neuromuscular fatigability is a crucial physical parameter of the daily living. The quantification of explosive force has been shown to be a sensitive means to assess neuromuscular fatigability. Thus, our study used explosive force estimates to assess neuromuscular fatigability in elderly CKD patients. METHODS: Inclusion criteria for CKD patients were age ≥ 60 years old and glomerular filtration rate (GFR) < 45 mL/min/1.73 m2 not on dialysis, and those for controls were GFR > 60 mL/min/1.73 m2 , age and diabetes matched. The fatigability protocol focused on a handgrip task coupled with surface electromyography (sEMG). Scalars were extracted from the rate of force development (RFD): absolute and normalized time periods (50, 75, 100, 150 and 200 ms, RFD50 , RFD75 , RFD100 , RFD150 and RFD200 , respectively), peak RFD (RFDpeak in absolute; NRFDpeak normalized), time-to-peak RFD (t-RFDpeak ) and the relative force at RFDpeak (MVF-RFDpeak ). A statistical parametric mapping approach was performed on the force, impulse and RFD-time curves. The integrated sEMG with time at 0-30, 0-50, 0-100 and 0-200 ms time intervals relative to onset of sEMG activity was extracted and groups were compared separately for each sex. RESULTS: The cohort of 159 individuals had a median age of 69 (9IQR ) years and body mass index was 27.6 (6.2IQR ) kg/m2 . Propensity-score-matched groups balanced CKD patients and controls by gender with 66 males and 34 females. In scalar analysis, CKD patients manifested a higher decrement than controls in the early phase of contraction, regarding the NRFDpeak (P = 0.009; η2 p = 0.034) and RFD75 and RFD100 (for both P < 0.001; η2 p = 0.068 and 0.064). The one-dimensional analysis confirmed that CKD males manifest higher and delayed neuromuscular fatigability, especially before 100 ms from onset of contraction. sEMG was lower in CKD patients than controls in the 0-100 ms (at rest: P = 0.049, Cohen's d = 0.458) and 0-200 ms (at rest: P = 0.016, Cohen's d = 0.496; during exercise: P = 0.006, Cohen's d = 0.421) time windows. Controls showed greater decrease of sEMG than CKD patients in the 0-30 ms (P = 0.020, Cohen's d = 0.533) and 0-50 ms (P = 0.010, Cohen's d = 0.640) time windows. As opposite to females, males showed almost the same differences between groups. CONCLUSIONS: Our study is the first to show that CKD patients have higher fatigability than controls, which may be associated with an impaired motor-unit recruitment, highlighting a neural drive disturbance with CKD. Further studies are needed to confirm these findings.
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Non-coding RNAs (sRNA) play a key role in controlling gene expression in bacteria, typically by base-pairing with ribosome binding sites to block translation. The modification of ribosome traffic along the mRNA generally affects its stability. However, a few cases have been described in bacteria where sRNAs can affect translation without a major impact on mRNA stability. To identify new sRNA targets in Bacillus subtilis potentially belonging to this class of mRNAs, we used pulsed-SILAC (stable isotope labeling by amino acids in cell culture) to label newly synthesized proteins after short expression of the RoxS sRNA, the best characterized sRNA in this bacterium. RoxS sRNA was previously shown to interfere with the expression of genes involved in central metabolism, permitting control of the NAD+/NADH ratio in B. subtilis. In this study, we confirmed most of the known targets of RoxS, showing the efficiency of the method. We further expanded the number of mRNA targets encoding enzymes of the TCA cycle and identified new targets. One of these is YcsA, a tartrate dehydrogenase that uses NAD+ as co-factor, in excellent agreement with the proposed role of RoxS in management of NAD+/NADH ratio in Firmicutes. IMPORTANCE Non-coding RNAs (sRNA) play an important role in bacterial adaptation and virulence. The identification of the most complete set of targets for these regulatory RNAs is key to fully identifying the perimeter of its function(s). Most sRNAs modify both the translation (directly) and mRNA stability (indirectly) of their targets. However, sRNAs can also influence the translation efficiency of the target primarily, with little or no impact on mRNA stability. The characterization of these targets is challenging. We describe here the application of the pulsed SILAC method to identify such targets and obtain the most complete list of targets for a defined sRNA.
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Bacillus subtilis , ARN Pequeño no Traducido , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , NAD/metabolismo , ARN Pequeño no Traducido/genética , ARN Pequeño no Traducido/metabolismo , ARN Bacteriano/metabolismo , ARN Mensajero/metabolismo , Regulación Bacteriana de la Expresión GénicaRESUMEN
INTRODUCTION: Chronic kidney disease (CKD) patients have a high degree of fatigue relating to neuromuscular symptoms. There is a lack of evidence regarding the etiology of neuromuscular fatigability in elderly CKD patients. METHODS: Inclusion criteria are as follows: age ≥60 yr, glomerular filtration rate (GFR) <45 mL·min -1 per 1.73 m 2 in CKD patients, and GFR >60 mL·min -1 ·1.73 m -2 in controls. The fatigability protocol consisted in a submaximal handgrip task at 40% peak force. Fatigue was assessed using the Multidimensional Fatigue Inventory-20 items (MFI-20) and the Functional Assessment of Chronic Illness Therapy-Fatigue questionnaires. Peak rate of force development (RFD peak , normalized: NRFD peak ) and rate of EMG rise (RER) were measured during explosive contractions; peak force and mean surface EMG were measured during maximum voluntary contractions. Multilevel models tested neuromuscular parameters adjusted for clinical and Multidimensional Fatigue Inventory-20 items subscales. Neuromuscular fatigability contribution to fatigue description was tested using model comparison. RESULTS: The study included 102 participants; 45 CKD patients and 57 controls. CKD mainly affected the mental and the reduced motivation subscales of fatigue. CKD was associated with greater neuromuscular fatigability assessed using NRFD peak (group-time interaction, -16.7 % MVF·s -1 , P = 0.024), which increased with fatigue severity ( P = 0.018) and with a higher rate of decrement in RER compared with controls (RER at 50 ms: ß = -121.2 µV·s -1 , P = 0.016, and ß = -48.5 µV·s -1 , P = 0.196, respectively). Furthermore, these patients show an association between the reduced motivation subscale and the RER (e.g., 30 ms: ß = -59.8% EMG peak ·s -1 , P < 0.001). Only peak force fatigability contributed to fatigue variance, whereas RFD peak did not. CONCLUSIONS: In CKD patients, the neuromuscular fatigability assessed using RFD peak is related to an impairment in motor-unit recruitment or discharge rates, whereas only peak force fatigability was related to fatigue. This suggests that targeting exercise interventions might lessen fatigue and improve quality of life in CKD patients.
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Fuerza de la Mano , Calidad de Vida , Humanos , Anciano , Electromiografía/métodos , Fatiga/etiología , Fatiga Muscular , Músculo EsqueléticoRESUMEN
BACKGROUND: In the general population, sport activity is associated with better health and better self-esteem. Among people living with HIV (PLHIV), sport activity could also be associated with better self-esteem. The main objective of our study was to assess the association between sport activity and self-esteem among people living with HIV. The secondary objectives were to evaluate the associations between sport activity with fatigue as well as with pain. METHODS: We performed a cross-sectional observational study among PLHIV in our region (Pays de la Loire in France). Each adult seen in routine HIV care was invited to participate in the study. Participants were invited to fill out self-questionnaires about sport activity, self-esteem, fatigue, and pain. The 2 groups of participants with and without sport activity were compared with a T Student test for self-esteem, fatigue, and pain scales. RESULTS: Among the 1160 people included in the study, 47% performed sport activity. The self-esteem score was better in the "sporting group" compared with the "non sporting group" (Rosenberg mean scale 32.7 ± 5.1/40 vs 31.9 ± 5 p = 0.01). The Functional Assessment of Chronic Illness Therapy Fatigue scale showed a lower fatigue in the sporting group than in the non-sporting group (mean total score 125 ± 22 vs 118 ± 24 p < 0.0001). The sporting group had a lower mean pain score (1.1 ± 1.8) than the non sporting group (1.4 ± 1.9 p = 0.004). CONCLUSIONS: Among PLHIV in our region, sport activity was associated with better self-esteem, lower fatigue and lower pain. Sport activity should be included in patient care for people living with HIV.
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Infecciones por VIH , Autoimagen , Adulto , Humanos , Estudios Transversales , Fatiga/etiología , Infecciones por VIH/epidemiología , DolorRESUMEN
Objective: Fingertip photoplethysmography (PPG) resulting from high-pass filtered raw PPG signal is often used to record arterial pulse changes in patients with suspected thoracic outlet syndrome (TOS). Results from venous (low-pass filtered raw signal) forearm PPG (V-PPG) during the Candlestick-Prayer (Ca + Pra) maneuver were recently classified into four different patterns in patients with suspected TOS, two of which are suggestive of the presence of outflow impairment. We aimed to test the effect of probe position (fingertip vs. forearm) and of red (R) vs. infrared (IR) light wavelength on V-PPG classification and compared pattern classifications with the results of ultrasound (US). Methods: In patients with suspected TOS, we routinely performed US imaging (US + being the presence of a positional compression) and Ca + Pra tests with forearm V-PPG IR . We recruited patients for a Ca + Pra maneuver with the simultaneous fingertip and forearm V-PPG R . The correlation of each V-PPG recording to each of the published pattern profiles was calculated. Each record was classified according to the patterns for which the coefficient of correlation was the highest. Cohen's kappa test was used to determine the reliability of classification among forearm V-PPG IR , fingertip V-PPG R , and forearm V-PPG R . Results: We obtained 40 measurements from 20 patients (40.2 ± 11.3 years old, 11 males). We found 13 limbs with US + results, while V-PPG suggested the presence of venous outflow impairment in 27 and 20 limbs with forearm V-PPG IR and forearm V-PPG R , respectively. Fingertip V-PPG R provided no patterns suggesting outflow impairment. Conclusion: We found more V-PPG patterns suggesting venous outflow impairment than US + results. Probe position is essential if aiming to perform upper-limb V-PPG during the Ca + Pra maneuver in patients with suspected TOS. V-PPG during the Ca + Pra maneuver is of low cost and easy and provides reliable, recordable, and objective evidence of forearm swelling. It should be performed on the forearm (close to the elbow) with either PPG R or PPG IR but not at the fingertip level.
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sRNAs are a taxonomically-restricted but transcriptomically-abundant class of post-transcriptional regulators. While of major importance for adaption to the environment, we currently lack global-scale methodology enabling target identification, especially in species without known RNA hub proteins (e.g. Hfq). Using psoralen RNA cross-linking and Illumina-sequencing we identify RNA-RNA interacting pairs in vivo in Bacillus subtilis, resolving previously well-described interactants. Although sRNA-sRNA pairings are rare (compared with sRNA-mRNA), we identify a robust example involving the conserved sRNA RoxS and an unstudied sRNA RosA (Regulator of sRNA A). We show RosA to be the first confirmed RNA sponge described in a Gram-positive bacterium. RosA interacts with at least two sRNAs, RoxS and FsrA. The RosA/RoxS interaction not only affects the levels of RoxS but also its processing and regulatory activity. We also found that the transcription of RosA is repressed by CcpA, the key regulator of carbon-metabolism in B. subtilis. Since RoxS is already known to be transcriptionally controlled by malate via the transcriptional repressor Rex, its post-transcriptional regulation by CcpA via RosA places RoxS in a key position to control central metabolism in response to varying carbon sources.
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Bacillus subtilis/genética , ARN Bacteriano/metabolismo , ARN Pequeño no Traducido/metabolismo , Bacillus subtilis/metabolismo , Proteínas Bacterianas/metabolismo , Carbono/metabolismo , Aptitud Genética , Proteoma , Procesamiento Postranscripcional del ARN , Estabilidad del ARN , ARN Pequeño no Traducido/biosíntesis , ARN Pequeño no Traducido/genética , ARN Pequeño no Traducido/fisiología , Transcripción GenéticaRESUMEN
In oxygen (O2) limiting environments, numerous aerobic bacteria have the ability to shift from aerobic to anaerobic respiration to release energy. This process requires alternative electron acceptor to replace O2 such as nitrate (NO3 -), which has the next best reduction potential after O2. Depending on the organism, nitrate respiration involves different enzymes to convert NO3 - to ammonium (NH4 +) or dinitrogen (N2). The expression of these enzymes is tightly controlled by transcription factors (TFs). More recently, bacterial small regulatory RNAs (sRNAs), which are important regulators of the rapid adaptation of microorganisms to extremely diverse environments, have also been shown to control the expression of genes encoding enzymes or TFs related to nitrate respiration. In turn, these TFs control the synthesis of multiple sRNAs. These results suggest that sRNAs play a central role in the control of these metabolic pathways. Here we review the complex interplay between the transcriptional and the post-transcriptional regulators to efficiently control the respiration on nitrate.
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The ways in which locations of ischemia and ischemic pain affect spatiotemporal gait parameters and leg electromyographic activity during walking have never been investigated in patients with peripheral arterial disease presenting intermittent claudication. Two groups were classified according to unilateral location of ischemia (distal, n = 10, or proximo-distal, n = 12). Patients described pain and three gait phases-initial pain-free, onset of pain and maximum pain-were analyzed. Patients with proximo-distal ischemia walked less (230 ± 111 m vs 384 ± 220 m), with increased step length, step time (+ 5.4% and + 5.8%) and reduced cadence (- 8.2%), than patients with distal ischemia. In both, the peaks of vertical ground reaction force were reduced in maximum pain (Peak1-distal: - 11.4%, Peak1-proximo-distal: - 10.3%; Peak2-distal: - 11.8%, Peak2-proximo-distal: - 9.0%). In the proximo-distal group, tibialis anterior activation peak and time were lower than in the distal group (- 4.5% and - 19.7%). During the maximum pain phase, this peak decreased only in the proximo-distal group (- 13.0%), and gastrocnemius medialis activation peak and time decreased in both groups (- 2.5% in distal and - 4.5% in proximo-distal). Thus, proximo-distal ischemia leads to more adverse consequences in gait than distal ischemia only. Increasing ischemic pain until maximum, but not onset of pain, induced gait adaptations.
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Claudicación Intermitente/fisiopatología , Músculo Esquelético/fisiología , Dolor/patología , Enfermedad Arterial Periférica/fisiopatología , Caminata , Adaptación Fisiológica , Anciano , Fenómenos Biomecánicos , Electromiografía , Humanos , Persona de Mediana Edad , Enfermedad Arterial Periférica/diagnósticoRESUMEN
BACKGROUND: We seek to evaluate whether ischemia extent (unilateral or bilateral) impacts spatiotemporal and neuromuscular gait parameters differently in patients with peripheral arterial disease and presenting intermittent claudication (PAD-IC). METHODS: Two groups of PAD-IC patients: unilateral (Unilat-IC; n = 15), bilateral (Bilat-IC; n = 15) and a group of control subjects with similar risk factors (n = 15) were evaluated during a constant load treadmill walking test. Spatiotemporal parameters and neuromuscular activation in tibialis anterior and gastrocnemius medialis were recorded. Patients were instructed to describe their pain during walking test, and three phases were analysed: pain-free, onset of pain and maximum pain in PAD-IC patients. FINDINGS: Single leg stance in the asymptomatic leg of Unilat-IC increases and becomes higher than the symptomatic leg and the Bilat-IC legs at maximum pain. Step time is higher and cadence is lower in PAC-IC than in controls. Tibialis anterior activation peak in Unilat-IC continuously decreases between phases and becomes lower than in Bilat-IC during maximum pain. Tibialis anterior activation time is higher in Bilat-IC and in the asymptomatic leg than in the symptomatic of Unilat-IC during all the phases. Gastrocnemius medialis activation peak in Bilat-IC decreases with pain. Gastrocnemius medialis activation time in the symptomatic leg of Unilat-IC presents a significant decrease between pain-free and maximum pain phases. INTERPRETATION: Ischemia impacts gait in PAD-IC patients differently according to its extent between legs compared to controls. Imbalance between legs in Unilat-IC induces compensatory mechanism and an asymmetrical pattern. Bilat-IC should not be simply considered as a 'double' Unilat-IC when evaluating gait.
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Marcha , Claudicación Intermitente , Fenómenos Biomecánicos , Humanos , Claudicación Intermitente/diagnóstico , Isquemia/diagnóstico , CaminataRESUMEN
Most bacterial ribonucleases (RNases) known to date have been identified in either Escherichia coli or Bacillus subtilis. These two organisms lie on opposite poles of the phylogenetic spectrum, separated by 1-3 billion years of evolution. As a result, the RNA maturation and degradation machineries of these two organisms have little overlap, with each having a distinct set of RNases in addition to a core set of enzymes that is highly conserved across the bacterial spectrum. In this paper, we describe what the functions performed by major RNases in these two bacteria, and how the evolutionary space between them can be described by two opposing gradients of enzymes that fade out and fade in, respectively, as one walks across the phylogenetic tree from E. coli to B. subtilis.
La plupart des ribonucléases (RNases) bactériennes connues à ce jour ont été identifiées chez Escherichia coli ou Bacillus subtilis. Ces deux organismes se trouvent aux pôles opposés du spectre phylogénétique, séparés par 13 milliards d'années d'évolution. Par conséquent, les mécanismes de maturation et de dégradation de l'ARN de ces deux organismes se chevauchent peu, chacun possédant un ensemble distinct de RNases en plus d'un ensemble coeur d'enzymes hautement conservées dans tout le spectre bactérien. Dans cet article, nous décrivons les fonctions remplies par les principales RNases de ces deux bactéries, et comment l'espace évolutif qui les sépare peut être décrit par deux gradients opposés d'enzymes qui disparaissent et apparaissent, respectivement, lorsqu'on parcourt l'arbre phylogénétique de E. coli à B. subtilis.
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Bacillus subtilis/enzimología , Bacillus subtilis/genética , Escherichia coli/enzimología , Escherichia coli/genética , Ribonucleasas , Escherichia coli/metabolismo , Filogenia , Ribonucleasas/química , Ribonucleasas/genética , Ribonucleasas/metabolismoRESUMEN
BACKGROUND: In patients with peripheral arterial disease and presenting intermittent claudication (PAD-IC), the pain due to ischemia impacts gait parameters, particularly in cases of unilateral disease. Deterioration of gait parameters in a pathological context is frequently associated with increased coactivation (simultaneous activation of agonist and antagonist muscles around a joint). RESEARCH QUESTION: Does unilateral PAD-IC affect the coactivation pattern during walking? Does the coactivation pattern change with increasing pain intensity? METHOD: We evaluated symptomatic and asymptomatic legs in 17 subjects with unilateral PAD-IC and 16 without PAD-IC (control group), during walking. Tibialis anterior (TA) and gastrocnemius medialis (GM) electromyographic activity, and peaks of vertical ground reaction force were recorded in this prospective study. We analyzed the coactivation index (CI(GM/TA)) during three periods (pain-free, pain and maximum pain) and phases of the gait cycle. Statistical analysis was carried out using the ANOVA procedure. RESULTS: During single support, CI(GM/TA) increases in the symptomatic leg during the pain period (+28 %) and in the asymptomatic leg during the maximum pain period (+29 %). During second double support, CI(GM/TA) increases in the symptomatic leg only (+49 %). In these gait phases, pain elicits differences in CI(GM/TA) between legs (pâ¯<â¯0.05). Second peak force decreases in the symptomatic leg only (-9%) and is negatively correlated with CI(GM/TA) during the three periods (r = -0.57; -0.76 and -0.78 respectively, pâ¯<â¯0.05). No difference is found in the control group. SIGNIFICANCE: The appearance and development of pain in the lower limbs is associated with a higher level of CI(GM/TA), revealing a compensatory gait pattern in PAD-IC patients. Optimal prevention, rehabilitation and re-training strategies for PAD-IC patients should take into consideration neuromuscular compensatory mechanisms between asymptomatic and symptomatic legs.
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Fenómenos Biomecánicos/fisiología , Claudicación Intermitente/fisiopatología , Pierna/fisiopatología , Caminata/fisiología , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios ProspectivosRESUMEN
Ribonucleases can cleave RNAs internally in endoribonucleolytic mode or remove one nucleotide at a time from either the 5' or 3' end through exoribonuclease action. To show direct implication of an RNase in a specific pathway of RNA maturation or decay requires the setting up of in vitro assays with purified enzymes and substrates. This chapter complements Chapter 24 on assays of ribonuclease action in vivo by providing detailed protocols for the assay of B. subtilis RNases with prepared substrates in vitro.
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Pruebas de Enzimas/métodos , Sondas ARN/metabolismo , ARN Bacteriano/metabolismo , Ribonucleasas/metabolismo , Bacillus subtilis/enzimología , Regulación Bacteriana de la Expresión Génica , CinéticaRESUMEN
Ribonucleases remodel RNAs to render them functional or to send them on their way toward degradation. In our laboratory, we study these pathways in detail using a plethora of different techniques. These can range from the isolation of RNAs in various RNase mutants to determine their implication in maturation or decay pathways by Northern blot, to proving their direct roles in RNA cleavage reactions using purified enzymes and transcribed substrates in vitro. In this chapter, we provide in-depth protocols for the techniques we use daily in the laboratory to assay RNase activity in vivo, with detailed notes on how to get these methods to work optimally. This chapter complements Chapter 25 on assays of ribonuclease action in vitro.