The effect of breaking up prolonged sitting on paired associative stimulation-induced plasticity.

Paired associative stimulation (PAS) can induce plasticity in the motor cortex, as measured by changes in corticospinal excitability (CSE). This effect is attenuated in older and less active individuals. Although a single bout of exercise enhances PAS-induced plasticity in young, physically inactive adults, it is not yet known if physical activity interventions affect PAS-induced neuroplasticity in middle-aged inactive individuals. Sixteen inactive middle-aged office workers participated in a randomized cross-over design investigating how CSE and short-interval intracortical inhibition (SICI) were affected by PAS preceded by 3 h of sitting (SIT), 3 h of sitting interrupted every 30 min by 3 min of frequent short bouts of physical activity (FPA) and 2.5 h of sitting followed by 25 min of moderate-intensity exercise (EXE). Transcranial magnetic stimulation was applied over the primary motor cortex (M1) of the dominant abductor pollicis brevis to induce recruitment curves before and 5 min and 30 min post-PAS. Linear mixed models were used to compare changes in CSE using time and condition as fixed effects and subjects as random effects. There was a main effect of time on CSE and planned within-condition comparisons showed that CSE was significantly increased from baseline to 5 min and 30 min post-PAS, in the FPA condition, with no significant changes in the SIT or EXE conditions. SICI decreased from baseline to 5 min post-PAS, but this was not related to changes in CSE. Our findings suggest that in middle-aged inactive adults, FPAs may promote corticospinal neuroplasticity. Possible mechanisms are discussed.

Strength training improves muscle aerobic capacity and glucose tolerance in elderly.

The primary aim of this study was to investigate the effect of short-term resistance training (RET) on mitochondrial protein content and glucose tolerance in elderly. Elderly women and men (age 71 ± 1, mean ± SEM) were assigned to a group performing 8 weeks of resistance training (RET, n = 12) or no training (CON, n = 9). The RET group increased in (i) knee extensor strength (concentric +11 ± 3%, eccentric +8 ± 3% and static +12 ± 3%), (ii) initial (0-30 ms) rate of force development (+52 ± 26%) and (iii) contents of proteins related to signaling of muscle protein synthesis (Akt +69 ± 20 and mammalian target of rapamycin +69 ± 32%). Muscle fiber type composition changed to a more oxidative profile in RET with increased amount of type IIa fibers (+26.9 ± 6.8%) and a trend for decreased amount of type IIx fibers (-16.4 ± 18.2%, P = 0.068). Mitochondrial proteins (OXPHOS complex II, IV, and citrate synthase) increased in RET by +30 ± 11%, +99 ± 31% and +29 ± 8%, respectively. RET resulted in improved oral glucose tolerance measured as reduced area under curve for glucose (-21 ± 26%) and reduced plasma glucose 2 h post-glucose intake (-14 ± 5%). In CON parameters were unchanged or impaired. In conclusion, short-term resistance training in elderly not only improves muscular strength, but results in robust increases in several parameters related to muscle aerobic capacity.

Quadriceps and hamstring muscle activity during cycling as measured with intramuscular electromyography.

PURPOSE: The aim of this study was to describe thigh muscle activation during cycling using intramuscular electromyographic recordings of eight thigh muscles, including the biceps femoris short head (BFS) and the vastus intermedius (Vint). METHODS: Nine experienced cyclists performed an incremental test (start at 170 W and increased by 20 W every 2 min) on a bicycle ergometer either for a maximum of 20 min or to fatigue. Intramuscular electromyography (EMG) of eight muscles and kinematic data of the right lower limb were recorded during the last 20 s in the second workload (190 W). EMG data were normalized to the peak activity occurring during this workload. Statistical significance was assumed at p ≤ 0.05. RESULTS: The vastii showed a greater activation during the 1st quadrant compared to other quadrants. The rectus femoris (RF) showed a similar activation, but with two bursts in the 1st and 4th quadrants in three subjects. This behavior may be explained by the bi-articular function during the cycling movement. Both the BFS and Vint were activated longer than, but in synergy with their respective agonistic superficial muscles. CONCLUSION: Intramuscular EMG was used to verify muscle activation during cycling. The activation pattern of deep muscles (Vint and BFS) could, therefore, be described and compared to that of the more superficial muscles. The complex coordination of quadriceps and hamstring muscles during cycling was described in detail.

Motor control of the trunk during a modified clean and jerk lift.

The purpose of the present study was to investigate the pattern of trunk muscle activation and intra-abdominal pressure (IAP) in a somewhat modified version of the clean and jerk lift. Nine healthy physically active male amateurs performed the exercise with a 30-kg barbell. Muscle activity was registered with electromyography from transversus abdominis (TrA) and obliquus internus (OI) using intramuscular electrodes and from rectus abdominis (RA) and erector spinae (ES) with surface electrodes. IAP was recorded with a nasogastric catheter. Measurements were made in various static positions throughout the lift and in the transitional phases separating them, both during lifting and lowering. The results demonstrated that the innermost abdominal muscle, TrA, showed increased activation levels in the two highest positions, whereas ES was most active, together with the highest IAP, in the lowest position. OI and RA showed generally little activation and no obvious trend throughout the lift. The results strengthen the view of a contributing role of TrA to the upright control of the trunk and indicate that the clean and jerk lift might constitute a whole-body exercise, still targeting the TrA muscle, in late-stage rehabilitation, especially for athletes during return to sports.

Concurrent EMG feedback acutely improves strength and muscle activation.

The purpose of this study was to investigate the acute effects of electromyographic (EMG) feedback on muscle activation and strength during maximal voluntary concentric and eccentric muscle actions. 15 females performed two sets of three lengthening and three shortening maximal voluntary isokinetic knee extensions at 20° s(-1) over 60° range of motion. After the first set, subjects were randomized to either a control group (n = 8) or a feedback group (n = 7). In the second set, the control group performed tasks identical to those in the first set, whereas the feedback group additionally received concurrent visual feedback of the EMGrms from Vastus Medialis (VM). Knee extensor strength and EMG activation of VM, Vastus lateralis (VL) and hamstrings (HAM) were measured during the MVCs. Analyses were performed separately in a 1 s preactivation phase, a 1 s initial movement phase and a 1 s late movement phase. EMG feedback was associated with significantly higher knee extensor strength in all phases (20.5% p < 0.05, 18.2% p < 0.001 and 19% p < 0.001, respectively) for the eccentric MVCs and in the preactivation phase (16.3%, p < 0.001) and initial movement phases (7.2%, p < 0.05) for concentric MVCs. EMG feedback from VM further improved activation in VM and HAM but not VL. These findings suggested that concurrent visual EMG feedback from VM could acutely enhance muscle strength and activation. Before recommending implementation of EMG feedback in resistance training paradigms, the feedback parameters needs to be optimized and its long-term effects needs to be scrutinized.

Downregulation of tenascin expression by glucocorticoids in bone marrow stromal cells and in fibroblasts.

Tenascin, a predominantly mesenchymal extracellular matrix (ECM) glycoprotein has a rather restricted tissue distribution, but until now factors that inhibit its expression have not been identified. Glucocorticoids are known to be beneficial for establishment of myelopoiesis in long-term bone marrow cultures. Tenascin was found to be expressed in the bone marrow, and glucocorticoids were found to affect bone marrow tenascin expression. Both tenascin mRNAs and the mRNA of another ECM protein, laminin B1 chain, were drastically downregulated by glucocorticoids during initiation of bone marrow cultures. However, in already established long-term cultures glucocorticoids did not affect laminin B1 chain mRNA levels although tenascin mRNAs continued to be downregulated. Studies with a stromal cell line (MC3T3-G2/PA6) and fibroblasts (3T3) suggested that glucocorticoids act directly on the stromal cells that produce tenascin. In 3T3 cells this downregulation occurred within 12 h of glucocorticoid-treatment, suggesting that glucocorticoids acted through cis regulatory elements of the tenascin gene. We suggest that glucocorticoids in part regulate hematopoiesis by modifying the ECM. Furthermore, downregulation of tenascin expression by glucocorticoids may in part explain the restricted tissue distribution of tenascin in other tissues.

A genomic clone encoding a novel proliferation-dependent histone H2A.1 mRNA enriched in the poly(A)+ fraction.

Replication-dependent histone mRNAs are prime examples of nonpolyadenylated mRNAs. We isolated and characterized cDNAs and a genomic clone for a replication-dependent histone H2A.1 mRNA which segregated into the poly(A)+ fraction during mRNA isolation through an oligo(dT)-cellulose column. However, the results of sequencing of the genomic clone suggested that the mRNA did not contain a poly(A) tail. Instead, the genomic sequence revealed a nonterminal oligo(A) tract directly upstream from the typical 3'-terminal hairpin loop of replication-dependent histone mRNAs. The nonterminal oligo(A) tract consisted of 14 adenylate residues interrupted by one guanylate residue (A4GA10). We concluded that this short oligo(A) stretch mediated binding of the mRNA to oligo(dT) even after stringent washes with 0.1 M NaCl, indicating that rather short oligo(A) sequences can ensure binding to oligo(dT)-cellulose. The cDNA and genomic clones contained an AAATAAG sequence at the end of the coding region. It has been suggested that this sequence contains a polyadenylation signal in some yeast and mouse transcripts, but it does not function as a polyadenylation signal in the histone transcript described in this paper.

Expansion of the nonadherent myeloid cell population by monoclonal antibodies against tenascin-C in murine long-term bone marrow cultures.

Tenascin-C, a predominantly mesenchymal extracellular matrix protein, has a restricted distribution in adult tissues. It has previously been shown that this protein is expressed in the bone marrow. In this paper we show that murine myeloid and lymphoid long-term bone marrow cultures differ in their expression of tenascin-C splice variants. In the adherent stromal layer of myeloid cultures, the 260-kDa polypeptide encoded by the 8-kb mRNA was the major splice variant, whereas in the stromal layer of lymphoid cultures both the shorter 210-kDa polypeptide encoded by the 6-kb mRNA and the 260-kDa polypeptide were abundantly expressed. However, in both culture systems the larger 260-kDa tenascin-C polypeptide was the major isoform secreted in the culture supernatant. This finding is in agreement with previous reports indicating that the smaller 210-kDa isoform is preferentially deposited in the stroma, whereas the alternatively spliced segment in the 260-kDa tenascin-C may contain anti-adhesive domains. Glucocorticoids in myeloid long-term bone marrow cultures and in the MC3T3-G2/PA6 cell line downregulated the expression of tenascin-C. In the present study we observed that this was due primarily to downregulation of the 8-kb major splice variant of the tenascin-C mRNA. We also studied the possible role of tenascin-C in the bone marrow by using antibodies against tenascin-C in long-term bone marrow cultures. We found that three monoclonal antibodies against the carboxyterminal type III fibronectin repeats of tenascin-C (TNCfn 7-8) increased the number of the non-adherent myeloid cells in myeloid long-term bone marrow cultures. It has recently been suggested that the TNCfn 6-8 domain of tenascin-C binds to the alpha8beta1 integrin. Using Northern blotting, we found that the integrin alpha8 subunit was expressed in adherent cells in bone marrow cultures, raising the possibility that tenascin-C acts in bone marrow cultures by binding to the alpha8beta1 integrin.

Trunk Muscle Activation at the Initiation and Braking of Bilateral Shoulder Flexion Movements of Different Amplitudes.

The aim of this study was to investigate if trunk muscle activation patterns during rapid bilateral shoulder flexions are affected by movement amplitude. Eleven healthy males performed shoulder flexion movements starting from a position with arms along sides (0°) to either 45°, 90° or 180°. EMG was measured bilaterally from transversus abdominis (TrA), obliquus internus (OI) with intra-muscular electrodes, and from rectus abdominis (RA), erector spinae (ES) and deltoideus with surface electrodes. 3D kinematics was recorded and inverse dynamics was used to calculate the reactive linear forces and torque about the shoulders and the linear and angular impulses. The sequencing of trunk muscle onsets at the initiation of arm movements was the same across movement amplitudes with ES as the first muscle activated, followed by TrA, RA and OI. All arm movements induced a flexion angular impulse about the shoulders during acceleration that was reversed during deceleration. Increased movement amplitude led to shortened onset latencies of the abdominal muscles and increased level of activation in TrA and ES. The activation magnitude of TrA was similar in acceleration and deceleration where the other muscles were specific to acceleration or deceleration. The findings show that arm movements need to be standardized when used as a method to evaluate trunk muscle activation patterns and that inclusion of the deceleration of the arms in the analysis allow the study of the relationship between trunk muscle activation and direction of perturbing torque during one and the same arm movement.

Expression of laminin alpha 1, alpha 5 and beta 2 chains during embryogenesis of the kidney and vasculature.

Laminins, found predominantly in basement membranes, are large glycoproteins consisting of different subsets of alpha, beta and gamma chain subunits. To resolve conflicting data in the literature concerning coexpression of alpha 1 and beta 2 chains, expression of alpha 1 chain was studied with two different antisera against the E3 fragment of laminin alpha 1 chain. Expression of the alpha 1 chain was seen in several types of epithelial basement membranes throughout development, but its expression in rat glomerular basement membranes and some other types of epithelial basement membranes occurred only during early stages of development. By contrast, beta 2 chains were detected by immunofluorescence only during advanced stages of glomerulogenesis and vascular development. By Northern and Western blots, beta 2 chains were detected somewhat earlier, but in situ hybridization revealed that beta 2 chain was also confined to vasculature during the earlier stages. It thus seems that, in the tissues studied here, the expression of alpha 1 and beta 2 chains was mutually exclusive. To explore whether the newly described alpha 5 chain is expressed in locations lacking alpha 1 chain, expression of alpha 5 chain was studied by Northern blots and in situ hybridization. The alpha 5 chain was not uniformly expressed in all embryonic epithelial cell types but was present mainly in epithelial sheets which produce very little alpha 1 chain. There also appeared to be a developmental trend, with alpha 1 chain appearing early and alpha 5 later, in maturing epithelial sheets. The alpha 5 chain could be a major alpha chain of the adult glomerular basement membrane.

Laminin isoforms and epithelial development.

Several different approaches suggest that basement-membrane assembly is important for epithelial development. Basement membranes contain isoforms of collagen IV, proteoglycans, and noncollagenous glycoproteins such as the laminins and nidogens. The expression and role of laminins for epithelial morphogenesis is reviewed. Laminins are large heterotrimeric proteins composed of alpha, beta, and gamma chains. Many major epithelial laminins and their receptors have been identified recently, and the extracellular protein-protein interactions that drive basement-membrane assembly are beginning to be understood. Three laminin alpha-chains are typically made by epithelial, alpha 1, alpha 3, and alpha 5. Three major epithelial heterotrimers can at present be distinguished--laminin-1 (alpha 1 beta 1 gamma 1), laminin-5 (alpha 3 beta 3 gamma 2), and laminin-10 (alpha 5 beta 1 gamma 1)--but other heterotrimers may exist in epithelia. Laminins containing either alpha 1 or alpha 3 chains are largely limited to epithelia, whereas the alpha 5 is also found in endothelial and muscle basement membranes, particularly in the adult. Some epithelial cell types express several laminin alpha-chains, so it is relevant to test how the different laminins affect epithelial cells. Laminins interact with integrin type of receptors on the cell surface, but binding to other proteins has also recently been demonstrated. Two important recent discoveries are the identification of dystroglycan as a major laminin receptor in muscle and epithelia, and nidogen as a high-affinity laminin-binding protein important for basement-membrane assembly. Antibody perturbation experiments suggest that these protein-protein interactions are important for epithelial morphogenesis.

Critical chromosome rearrangement in acute promyelocytic leukemia.

We report here a patient with acute promyelocytic leukemia (APL) who has two normal chromosomes #15 but a structurally abnormal chromosome #17. This case indicates that the critical point of rearrangement in APL is not necessarily in chromosome #15 but may, alternatively, be in chromosome #17.

Chromosome abnormalities in 16 Finnish patients with Burkitt's lymphoma or L3 acute lymphocytic leukemia.

Eleven patients with Burkitt's lymphoma (BL), i.e., small noncleaved non-Hodgkin's lymphoma, and 5 patients with Burkitt-type acute lymphocytic leukemia (ALL-L3) were selected for chromosome study. Two of the 16 patients had no B-cell markers, but the erythrocyte marker--glycophorin A--was present on the surface of the leukemic blasts. The critical breakpoint at 8q24 was detected in 14 of the 16 patients, whereas this aberration was not detected in any of the 134 patients belonging to other subgroups of non-Hodgkin's lymphoma or ALL that we studied during the same period. In addition to the t(8;14)(q24;q32), the following translocations with the breakpoint at 8q24 were seen: t(2;8)(p11;q24), t(8;11)(q24;q13) in BL, and t(2;8;14)(p11 or p12;q24;q32) in ALL. Additional aberrations seen more than once were trisomy #7 and abnormalities in chromosomes #1, #11, and #13.

Estimation of unbound concentrations of morphine from microdialysate concentrations by use of nonlinear regression analysis in vivo and in vitro during steady state conditions.

The unbound concentration of morphine in striatum of rats was estimated during a constant rate infusion of morphine 14 mumol/h*kg, by use of the microdialysis technique and nonlinear regression analysis. The concentrations in plasma of morphine and its metabolite, morphine-3-glucuronide, were 4.2 +/- 1.4 microM and 7.7 +/- 4.0 microM, respectively, during the constant rate infusion. The corresponding estimated unbound concentrations of morphine in striatum varied between 0.06 and 0.11 microM. No morphine-3-glucuronide was detected in the brain dialysates. The unbound concentration in striatum was lower than expected based on unbound plasma concentrations and could be an indication of active transport from the brain. Five different equations were tested to find the best empirical description of the relationship between microdialysate concentration and perfusion rate by nonlinear regression analysis. The equations were validated by a serum in vitro study, where three unbound concentrations of morphine estimated from microdialyis were compared to estimates obtained from equilibrium dialysis. The precision of the parameter estimates obtained from the five equations was tested by Monte Carlo simulations. One of the equations (Eq. 4) was selected in preference to the others, because of the good agreement with the estimated unbound concentration obtained by equilibrium dialysis in vitro, and good precision of the parameter estimates. The method described in this paper is valuable when estimating the unbound concentration of drug from microdialysate concentrations during steady state conditions. Furthermore, the method is easily accessible when working in the pharmacokinetic and pharmacodynamic field.

Activation of transversus abdominis varies with postural demand in standing.

Transversus abdominis (TrA) is a multifunctional muscle, being involved in pressure regulation within the abdominal cavity and thereby in direction independent stabilization of the spine and resistance to imposed trunk flexion moments. Indirect evidence suggests a role of TrA also in postural control of the erect human trunk. The main purpose here was to investigate if the magnitude of TrA activation is related to postural demand. Eleven healthy males performed seven different symmetrical static bilateral arm positions holding 3 kg in each hand. The arm positions were selected to systematically vary the height of the centre of mass (COM) keeping imposed moments constant and vice versa. EMG was recorded bilaterally with fine-wire intramuscular electrodes from TrA and obliquus internus (OI) and with surface electrodes from rectus abdominis (RA) and erector spinae (ES). Intra-abdominal pressure (IAP) was measured via a pressure transducer in the gastric ventricle. TrA was the only muscle that displayed activation co-varying with the vertical position of the COM. Further, TrA activation increased, together with IAP and ES activation, with imposed flexion moment, i.e. with arms extended horizontally forward. In contrast to OI, RA and ES, TrA activation was independent of the direction of the imposed moment (arms held inclined forward or backward). In conclusion, TrA activation level is uniquely associated with increased postural demand caused by elevated COM. Also, TrA appears to assist in counteracting trunk flexion via increased IAP, and contribute to general spine stabilization when the trunk is exposed to moderate flexion and extension moments.

Expansion of highly differentiated CD8+ T-cells or NK-cells in patients treated with dasatinib is associated with cytomegalovirus reactivation.

The tyrosine kinase inhibitor dasatinib exerts immunosuppressive effects on T-cells and NK-cells in vitro. However, in some dasatinib-treated leukemia patients, clonal lymphocytosis with large granular lymphocyte (LGL) morphology develops, and this is associated with enhanced therapeutic responses. To elucidate the mechanistic basis for this paradoxical observation, we conducted detailed phenotypic and functional analyses of T-cell and NK-cell populations from 25 dasatinib-treated leukemia patients. All tested patients with LGL expansions (15/16) were cytomegalovirus (CMV) immunoglobulin (IgG) seropositive with high frequencies of CMV-specific CD8(+) T-cells; 5/16 LGL patients also experienced symptomatic CMV reactivation during dasatinib therapy. Expanded T-cell and NK-cell populations exhibited late differentiated (CD27(-)CD57(+)) phenotypes; this was associated with a predisposition to apoptosis within the T-cell compartment and impaired NK-cell cytotoxicity. Only 3/9 non-LGL patients were CMV IgG seropositive. Dasatinib inhibited in vitro lymphocyte functions, similarly in LGL patients and controls. Notably, distinct CD8(high) and CD8(low) T-cell subsets were observed in LGL patients; this phenotypic dichotomy was also apparent in CMV-specific CD8(+) T-cell populations, and exhibited features consistent with antigen-driven activation. In addition, plasma levels of IP-10, IL-6, monokine induced by interferon-γ and interleukin-2R were significantly increased in LGL patients. These data provide evidence that dasatinib-associated LGL expansion is linked to CMV reactivation and suggest a potential mechanism for this phenomenon.

Improvements in dynamic plantar flexor strength after resistance training are associated with increased voluntary activation and V-to-M ratio.

The aim of this study was to investigate if, and via what mechanisms, resistance training of the plantar flexor muscles affects voluntary activation during maximal voluntary eccentric and concentric muscle actions. Twenty healthy subjects were randomized into a resistance training group (n = 9) or a passive control group (n = 11). Training consisted of 15 sessions of unilateral mainly eccentric plantar flexor exercise over a 5-wk period. During pre- and posttraining testing, dynamic plantar flexor strength was measured and voluntary activation was calculated using the twitch interpolation technique. The soleus Hoffman reflex (H-reflex) was used to assess motoneurone excitability and presynaptic inhibition of Ia afferents, whereas the soleus V-wave was used to test for changes in both presynaptic inhibition of Ia afferents and supraspinal inputs to the motoneurone pool. H-reflexes, V-waves, supramaximal M-waves, and twitches were evoked as the foot was moved at 5 degrees /s through an angle of 90 degrees during passive ankle rotations (passive H-reflexes and M-waves) and during maximal voluntary concentric and eccentric plantar flexions [maximal voluntary contraction (MVC) H-reflexes, M-waves, and V-waves]. Training induced significant improvements in plantar flexor strength and voluntary activation during both concentric and eccentric maximal voluntary actions. Soleus passive and MVC H-to-M ratios remained unchanged after training, whereas the soleus V-to-M ratio was increased during both concentric and eccentric contractions after training. No changes were found in the control group for any of the parameters. The enhanced voluntary strength could be attributed partly to an increase in voluntary activation induced by eccentric training. Since the passive and MVC H-to-M ratios remained unchanged, the increase in activation is probably not due to decreased presynaptic inhibition. The increased V-to-M ratio for both action types indicates that increased voluntary drive from supraspinal centers and/or modulation in afferents other than Ia afferents may have contributed to such an increase in voluntary activation.

Clonal expansion of T/NK-cells during tyrosine kinase inhibitor dasatinib therapy.

Dasatinib, a broad-spectrum tyrosine kinase inhibitor (TKI), predominantly targets BCR-ABL and SRC oncoproteins and also inhibits off-target kinases, which may result in unexpected drug responses. We identified 22 patients with marked lymphoproliferation in blood while on dasatinib therapy. Clonality and immunophenotype were analyzed and related clinical information was collected. An abrupt lymphocytosis (peak count range 4-20 x 10(9)/l) with large granular lymphocyte (LGL) morphology was observed after a median of 3 months from the start of therapy and it persisted throughout the therapy. Fifteen patients had a cytotoxic T-cell and seven patients had an NK-cell phenotype. All T-cell expansions were clonal. Adverse effects, such as colitis and pleuritis, were common (18 of 22 patients) and were preceded by LGL lymphocytosis. Accumulation of identical cytotoxic T cells was also detected in pleural effusion and colon biopsy samples. Responses to dasatinib were good and included complete, unexpectedly long-lasting remissions in patients with advanced leukemia. In a phase II clinical study on 46 Philadelphia chromosome-positive acute lymphoblastic leukemia, patients with lymphocytosis had superior survival compared with patients without lymphocytosis. By inhibiting immunoregulatory kinases, dasatinib may induce a reversible state of aberrant immune reactivity associated with good clinical responses and a distinct adverse effect profile.