Leg general muscle moment and power patterns in able-bodied subjects during recumbent cycle ergometry with ankle immobilization.

Rehabilitation of persons with pareses commonly uses recumbent pedalling and a rigid pedal boot that fixes the ankle joint from moving. This study was performed to provide general muscle moments (GMM) and joint power data from able-bodied subjects performing recumbent cycling at two workloads. Twenty-six able-bodied subjects pedalled a stationary recumbent tricycle at 60 rpm during passive cycling and at two workloads (low 15 W and high 40 W per leg) while leg kinematics and pedal forces were recorded. GMM and power were calculated using inverse dynamic equations. During the high workload, the hip and knee muscles produced extensor/flexor moments throughout the extensions/flexions phases of the joints. For low workload, a prolonged (crank angle 0-258°) hip extension moment and a shortened range (350-150°) of knee extension moment were observed compared to the corresponding extension phases of each joint. The knee and hip joints generated approximately equal power. At the high workload the hip and knee extensors generated increased power in the propulsion phase. For the first time, this study provides GMM and power patterns for able-bodied subjects performing recumbent cycling with an immobilized ankle. The patterns showed greater similarities to upright cycling with a free ankle, than previously supposed.

Comparison of the pedalling performance induced by magnetic and electrical stimulation cycle ergometry in able-bodied subjects.

The purpose of the study was to compare the mechanical power and work generated by able-bodied subjects during functional magnetic stimulation (FMS) vs. functional electrical stimulation (FES) induced ergometer training conditions. Both stimulation methods were applied at a 30 Hz frequency to the quadriceps muscles of 22 healthy able-bodied subjects to induce cycling for 4× four minutes or until exhaustion. FMS was performed via large surface, cooled coils, while FES was applied with a typical stimulation setup used for cycling. Significantly more (p<10(-3)) muscular power was generated by FMS (23.8 ± 9.1W [mean ± SD]) than by FES (11.3 ± 11.3 W). Additionally, significantly more (p<10(-6)) work was produced by FMS than by FES (4.413 ± 2.209 kJ vs. 0.974 ± 1.269 kJ). The increase in the work was paralleled by a significant prolongation of time to cycling failure (181.8 ± 33.4s vs. 87.0 ± 54.0 s, respectively, p<10(-5)). Compared to FES, FMS can produce more intense and longer cycling exercise in able-bodied subjects. The differing dynamic behaviour of FMS and FES in the presented measurement setup might be related to stimulation induced pain and fatigue mechanisms of the neuromuscular system.

Spinal and cortical activity-dependent plasticity following learning of complex arm movements in humans.

Activity-dependent plasticity is a fundamental requirement for human motor learning, which takes place at several stages of the motor system and involves various mechanisms in neuronal circuitry. Here, we investigate parameters of cortical and spinal excitability before and immediately after a single session of locomotion-like arm training (LMT) or sequential visuo-motor learning (VMT). Both training paradigms focused especially on mainly activating the flexor carpi radialis muscle (FCR). The activity-dependent change in the excitability of FCR-associated neurons was investigated using standard transcranial magnetic stimulation, including analysis of motor-evoked potentials (MEP) amplitude, short-interval intracortical inhibition and intracortical facilitation (ICF). Furthermore, spinal plasticity was also assessed by means of homosynaptic FCR H-reflex depression (HD). LMT decreased HD and ICF. In contrast, VMT had no significant effect on cortical and spinal parameters. There was a nonsignificant tendency of an increase in MEP amplitudes after both interventions. This implies that human locomotor-related learning involves spinal mechanisms. Despite the decreasing importance of quadrupedal coordination in the course of evolution, these changes in transsynaptic plasticity may reflect a persisting locomotor memory-encoding function in the spinal circuitry of the human upper extremities. Evaluating FCR HD might be helpful for the evaluation and development of locomotor rehabilitation strategies.

[Functional magnetic stimulation as a supposedly 'painless' option for movement induction in plegics]. / Funktionelle Magnetstimulation als "schmerzlose" Variante der künstlichen Bewegungsinduktion bei Lähmungen.

BACKGROUND: It is known in the rehabilitation of central pareses that functional electrical stimulation (FES) of the muscles can induce movement and accomplish training in patients. The main limitations of this method are that patients with preserved sensation experience pain and the reflexes triggered by FES. Therefore the application of the largely "painless" magnetic stimulation (FMS) of the muscles would be a potential alternative in the rehabilitation of patients with partially preserved sensation. As the generation of high force and power levels is considered to be an essential requirement of effective rehabilitation strategies, we have shown in previous work that FMS with large surface magnetic coils fitted to the thigh can generate about 2.5 times higher isometric forces in patients with preserved sensation, than can FES. OBJECTIVES: The goal of the present pilot study was to prove that the mechanical power generated by functional magnetic stimulation is superior to that produced by electrical stimulation too. METHODS: We have measured the mechanical torque, the power, the accomplished work and the kinematics in 4 healthy control subjects, who performed pedalling propelled by FMS and FES until complete muscular exhaustion, using a cycling test-bed under isotonic conditions (constant resistance). RESULTS: We have proved that the generated work, mean power, cadence and smoothness of pedalling essentially depend on peak torque and power. Furthermore, we found evidence that smoother pedalling could be achieved using magnetic, compared to electrical stimulation because of the higher peak torques that were generated by FMS. CONCLUSION: This study supports the concept that peripheral magnetic stimulation is an appropriate rehabilitation method for patients with central pareses and preserved sensory apparatus because FMS is less painful than electrical stimulation.

Force-pain relationship in functional magnetic and electrical stimulation of subjects with paresis and preserved sensation.

OBJECTIVE: Using "painless" magnetic stimulation (FMS) to support the cycling of paretic subjects with preserved sensation is possible and potentially superior to electrical stimulation (FES). We investigated the dependence of the torque and the pain evoked by FMS and FES on stimulation conditions in order to optimize magnetic stimulation. METHODS: Torque and pain induced by quadriceps stimulation in 13 subjects with paresis and preserved sensation (due to multiple sclerosis) were compared under the conditions: (1) small vs large stimulated surfaces of the thigh, (2) varying contraction velocities of the muscle (isometric vs 15 and 30 rpm isokinetic speed), (3) FMS vs FES modalities, and (4) varying magnetic coil locations. RESULTS: Torque and pain significantly depended on the amount of surface and location of stimulation during FMS, on the stimulation modality, and on the muscle contraction velocity during FES and FMS. FMS with a saddle-shaped coil produced more torque (p<0.05) than any other stimulation modality, even at 30 rpm velocity. CONCLUSIONS: To support leg cycling of subjects with preserved sensation, the application of FMS stimulation with a large-surface saddle-shaped coil and the focusing of stimulation on the lateral-frontal surface of the thigh produces greater torque and less pain than FES. SIGNIFICANCE: Optimized magnetic stimulation is a superior alternative to electrical stimulation in the rehabilitation of subjects with preserved sensation.

Fusogenic membrane glycoproteins induce syncytia formation and death in vitro and in vivo: a potential therapy agent for lung cancer.

Fusogenic membrane glycoproteins (FMGs) are viral envelope proteins, which bind surface receptors and induce fusion of the cell membrane. An FMG-transfected cell will fuse with neighbor cells, thus forming syncytia that die within 5 days. In this report, plasmids encoding for FMGs from Human Endogenous Retrovirus-W (HERV-W) was compared with Gibbon Ape Leukemia Virus (GALV) and feline endogenous virus RD-114 (RD). These plasmids were transfected in human non-small-cell lung cancer (NSCLC) cells in vitro or directly injected into tumors in mice. All FMGs induced the formation of syncytia containing around 50 cells. HERV-W or GALV FMGs decreased up to 80% of cell viability in vitro and inhibited tumor growth in vivo (60-70% reduction). In contrast, RD FMG was not efficient. Apoptosis played a role in the death of the syncytia, but addition of the caspase inhibitor Z-VAD-fmk had no effect, suggesting that apoptosis is not the only mechanism responsible for FMG-induced cell death. Altogether, our results demonstrate that even at very low transfection efficiency, the antitumor activity of HERV-W FMG is as effective as that of GALV in vitro and in vivo for the treatment of human lung tumors.

FES-propelled cycling of SCI subjects with highly spastic leg musculature.

BACKGROUND: The presence of spasms precludes the use of artificial electrical activation of the muscles to restore mobility. The prospect of using an electrical stimulus that produces motor activation without causing unwanted reflex activation in patients with high levels of spasticity is an appealing one. OBJECTIVE: The purpose of the study was to determine the efficacy of modulated middle frequency alternating current (MFAC) muscle stimulation compared to the conventional method of standard low frequency rectangular pulse (LFRP) stimulation used in cycling of persons with spinal cord injury (SCI) and pronounced spasticity. METHODS: To evaluate cycling-relevant differences between stimulation modes, 13 subjects with SCI (ASIA-A), 11 of them with strong spasticity, underwent isometric and cycling measurements using both 20 Hz LFRP and 4 KHz modulated with 50 Hz MFAC. The isometric long-lasting reflex torque response in the quadriceps and hamstrings muscles, and the dynamic work during 1000 sec of ergometric cycling as well as the number of involuntary stops caused by hyperreflexia were recorded. RESULTS: The long-lasting reflex torque response was significantly lower when using MFAC than with LFRP stimulation. During MFAC stimulation work generated was on average 374% higher (p = 0.002) and the number of involuntary stops was on average 32% lower (p < 0.001) than during standard LFRP stimulation-propelled cycling. CONCLUSION: These findings suggest that MFAC-stimulated cycling of strongly spastic SCI subjects is more effective in terms of generated isometric torque and power than stimulation with LFRP. Thus, more health benefits, e.g., cardiovascular and muscular training and spasticity-decreasing effects, can be expected faster using MFAC instead of LFRP in stimulation-propelled cycling.

Functional electrical stimulation assisted cycling of patients with subacute stroke: kinetic and kinematic analysis.

BACKGROUND: Cycling is a safe and functionally effective exercise for patients with early post-stroke and poor balance. Such exercise is considered even more effective when functional electrical stimulation is added. Our principal aim was to determine the biomechanically quantifiable parameters of cycling that can be improved in patients with subacute hemiparesis by incorporating functional electrical stimulation. These parameters were defined as objective goals that can be achieved in clinical applications. A secondary aim was to determine whether they could be used to identify subjects who would benefit from such therapy. METHODS: Using a tricycle testbed, we tested 39 subacute (mean 10.9 weeks post-stroke (SD 5.9)), hemiplegic subjects. During isometric measurements we recorded volitional and electrically evoked crank torques, the latter at maximal tolerable intensity. During ergometric measurements, volitional pedaling was alternated with combined pedaling (volitional supported by stimulation), performed at 30-s intervals. Power, smoothness, and symmetry of cycling were evaluated. FINDINGS: Twenty-six percent of the subjects significantly improved the smoothness of their cycling with functional electrical stimulation. Only 8% and 10% significantly increased their power and symmetry, respectively. The improvement in smoothness significantly correlated with the capability of the individual to generate electrical torque (Spearman's rank correlation coefficient=0.66 at P=0.001). INTERPRETATION: The smoothness of cycling was the most sensitive parameter improved by functional electrical stimulation. This improvement depended on the amount of torque evoked, and the torque achieved, in turn, correlated with the tolerated intensity of stimulation.

[The electrical stimulation bicycle: a neuroprosthesis for the everyday use of paraplegic patients]. / Das Elektrostimulationsfahrrad als Neuroprothese. Querschnittgelähmte Patienten mit dem Fahrrad unterwegs.

Until recently, few patients with complete paraplegia could walk or stand with the help of functional electrical stimulation (FES) of the leg muscles regularly at home. In comparison, FES cycling with an adapted tricycle is easy to put into practice because the legs remain connected to the pedals and through the use of a tricycle or stationary bicycle, the balancing problems of the patient recedes into the background. In the first German feasibility studies for paraplegic cycling, eleven completely paraplegic patients have been tested so far. The goal is to make FES cycling a daily activity in the lives of as many patients as possible.

[Paraplegic cycling using functional electrical stimulation. Experimental and model-based study of power output]. / Fahrradfahren Querschnittgelähmter mittels funktioneller Elektrostimulation. Eine experimentelle und modellbasierte Untersuchung der Leistungsabgabe.

Cycling using functional electrical stimulation offers paraplegics the possibility of muscle and cardiovascular training as well as the chance for independent locomotion. To investigate whether this method might be suitable for a large group of paraplegics, the first German feasibility study of functional electrical stimulation (FES) cycling with seven paraplegic patients was started at the beginning of 2003. Even at the beginning of the study, and without training, these patients were able to drive distances of 0.5-1.6 km. To stimulate cardiovascular adaptation processes in the case of FES ergometer training or to cover useful distances in the case of FES cycling, a minimum amount of generated mechanical output power is required, which as a rule cannot be achieved yet. In this study, we point out two particular aspects of FES cycling, which impair power output: prolonged fatigue mode and viscous joint friction of the paraplegic FES cyclist. We discuss current possibilities for increasing output power and endurance.

The cellular electrophysiologic effect of a new amiodarone like antiarrhythmic drug GYKI 16638 in undiseased human ventricular muscle: comparison with sotalol and mexiletine.

The cellular electrophysiologic effect of GYKI 16638, a new antiarrhythmic compound was studied and compared with that of sotalol and mexiletine in undiseased human right ventricular muscle preparation by applying the conventional microelectrode technique. GYKI 16638 (5 microM), at stimulation cycle length of 1000 ms, lengthened action potential duration (APD(90)) from 338.9 +/- 28.6 ms to 385.4 +/- 24 ms (n = 9, p > 0.05). This APD lengthening effect, unlike that of sotalol (30 microM), was rate-independent. GYKI 16638, contrary to sotalol and like mexiletine (10 microM), exerted a use-dependent depression of the maximal rate of depolarization (V(max)) which amounted to 36.4 +/- 11.7% at cycle length of 400 ms (n = 5, p < 0.05) and was characterised with an offset kinetical time constant of 298.6 +/- 70.2 ms. It was concluded that GYKI 16638 in human ventricular muscle shows combined Class IB and Class III antiarrhythmic properties, resembling the electrophysiological manifestation seen after chronic amiodarone treatment.

The slow component of the delayed rectifier potassium current in undiseased human ventricular myocytes.

OBJECTIVE: The purpose of this study was to investigate the properties of the slow component of the delayed rectifier potassium current (I(Ks)) in myocytes isolated from undiseased human left ventricles. METHODS: The whole-cell configuration of the patch-clamp technique was applied in 58 left ventricular myocytes from 15 hearts at 37 degrees C. Nisoldipine (1 microM) was used to block inward calcium current (I(Ca)) and E-4031 (1-5 microM) was applied to inhibit the rapid component of the delayed rectifier potassium current (I(Kr)). RESULTS: In 31 myocytes, an E-4031 insensitive, but L-735,821 and chromanol 293B sensitive, tail current was identified which was attributed to the slow component of I(K) (I(Ks)). Activation of I(Ks) was slow (tau=903+/-101 ms at 50 mV, n=14), but deactivation of the current was relatively rapid (tau=122.4+/-11.7 ms at -40 mV, n=19). The activation of I(Ks) was voltage independent but its deactivation showed clear voltage dependence. The deactivation was faster at negative voltages (about 100 ms at -50 mV) and slower at depolarized potentials (about 300 ms at 0 mV). In six cells, the reversal potential was -81.6+/-2.8 mV on an average which is close to the K(+) equilibrium potential suggesting K(+) as the main charge carrier. CONCLUSION: In undiseased human ventricular myocytes, I(Ks) exhibits slow activation and fast deactivation kinetics. Therefore, in humans I(Ks) differs from that reported in guinea pig, and it best resembles I(Ks) described in dog and rabbit ventricular myocytes.

Effects of endothelin-1 on calcium and potassium currents in undiseased human ventricular myocytes.

Endothelins have been reported to exert a wide range of electrophysiological effects in mammalian cardiac cells. These results are controversial and human data are not available. Our aim was to study the effects of endothelin-1 (ET-1, 8 nmol/l) on the L-type calcium current (ICa-L) and various potassium currents (rapid component of the delayed rectifier, IKr; transient outward current, Ito; and the inward rectifier K current, IK1) in isolated human ventricular cardiomyocytes. Cells were obtained from undiseased donor hearts using collagenase digestion via the segment perfusion technique. The whole-cell configuration of the patch-clamp technique was applied to measure ionic currents at 37 degrees C. ET-1 significantly decreased peak ICa-L from 10.2+/-0.6 to 6.8+/-0.8 pA/pF at +5 mV (66.7% of control, P<0.05, n=5). This reduction of peak current was accompanied by a lengthening of inactivation. The voltage dependence of steady-state activation and inactivation was not altered by ET- 1. IKr, measured as tail current amplitudes at 40 mV, decreased from 0.31+/-0.02 to 0.06+/-0.02 pA/pF (20.3% of control, P<0.05, n=4) after exposure to ET-1. ET-1 failed to change the peak amplitude of Ito, measured at +50 mV (9.3+/-4.6 and 9.0+/-4.4 pA/pF before and after ET-1, respectively), or steady-state IK1 amplitude, measured at the end of a 400-ms hyperpolarization to -100 mV (3.6+/-1.4 and 3.7+/-1.4 pA/pF, n=4). The present results indicate that in undiseased human ventricular myocytes ET-1 inhibits both ICa-L and IKr; however, the degree of suppression of the two currents is different.

[Risk assessment and measuring the need for revascularization on the basis of residual ischemia by means of dobutamine stress echocardiography in the early phase of the first myocardial infarction]. / A kockázat meghatározása és a várható revascularisatiós igény felmérése residualis ischaemia kimutatása alapján az elsö acut myocardialis infarctus korai fázisában dobutamin stressz echocardiographia segítségével.

The aim of this study is to present a modern method which can be used in the early phase of myocardial infarction. Its results show that with the detection of residual ischaemia, valuable information can be gained concerning the progression of the disease and the claim to revascularisation. During the first week (2-7 days), the authors performed risk assessing dobutamine stress echocardiography in 52 nonselected acute patients with first myocardial infarction. Patients with negative results were regarded as low risk cases 35/52 (67.3%), while those with positive results were regarded as high risk cases 17/52 (32.7%). The number of the serious postinfarction hard events was 22/52 (42.3%) during the one-year following period. In the case of the positive group, during mainly the first three months the occurrence of such events was found to be extremely high: 16/17 (94.1%), while in the case of the negative group it was only 6/35 (17.1%). On the basis of the test results it can be stated that coronarography was performed in 14/17 (82.4%) vs. 3/35 (8.6%) of the cases, and revascularisation in 10/17 (58.8%) vs. 1/35 (2.9%) of the cases. In 2/17 (11.8%) vs. 3/35 (8.6%) of the cases, the patients suffered from acute coronary syndrome and the mortality rate was 2/17 (11.8%) vs. 1/35 (2.9%). The sensitivity value was 79.1%, the specificity value 90.6%, and the positive and negative predictive values 86.3% and 85.3%. There was no such significant complication detected which could be attributable to the examination. On the basis of the results and corresponding to the data in special literature, the authors found the dobutamine stress test a good and safe prognostical method even some days after the infarction. With the help of this method the high and low risk groups of patients can be separated early, and at the same time the claim to revascularisation in the first three months can be measured too.

18F-FDG PET studies in patients with extratemporal and temporal epilepsy: evaluation of an observer-independent analysis.

UNLABELLED: The aim of this study was to evaluate an observer-independent analysis of 18F-fluorodeoxyglucose (FDG) PET studies in patients with temporal or extratemporal epilepsy. METHODS: Twenty-seven patients with temporal epilepsy and 22 patients with extratemporal epilepsy were included in the study. All patients with temporal epilepsy and 7 patients with extratemporal epilepsy underwent surgical treatment. In patients who showed significant postoperative improvement (temporal, n = 23; extratemporal, n = 6), the epileptogenic focus was assumed to be located in the area of surgical resection. In extratemporal epilepsy patients who did not undergo surgery, the focus localization was determined using a combination of semiology, ictal and interictal electroencephalography, [99mTc]ethyl cysteinate dimer SPECT, MRI and [11C]flumazenil PET. Visual analysis was performed by two experienced and two less experienced blinded observers using sagittal, axial and coronal images. In the automated analysis after anatomic standardization and generation of three-dimensional stereotactic surface projections (SSPs), a pixelwise comparison of 18F-FDG uptake with an age-matched reference database (n = 20) was performed, resulting in z score images. Pixels with the maximum deviation were detected, summarized and attached to one of 20 predefined surface regions of interest. For comparison with 18F-FDG PET and MR images, three-dimensional overlay images were generated. RESULTS: In patients with temporal epilepsy, the sensitivity was comparable for visual and observer-independent analysis (three-dimensional SSP 86%, experienced observers 86%-90%, less experienced observers 77%-86%). In patients with extratemporal epilepsy, three-dimensional SSP showed a significantly higher sensitivity in detecting the epileptogenic focus (67%) than did visual analysis (experienced 33%-38%, each less experienced 19%). In temporal lobe epilepsy, there was moderate to good agreement between the localization found with three-dimensional SSP and the different observers. In patients with extratemporal epilepsy, there was a high interobserver variability and only a weak agreement between the localization found with three-dimensional SSP and the different observers. Although three-dimensional SSP detected multiple lesions more often than visual analysis, the determination of the highest deviation from the reference database allowed the identification of the epileptogenic focus with a higher accuracy than subjective criteria, especially in extratemporal epilepsy. CONCLUSION: Three-dimensional SSP increases sensitivity and reduces observer variability of the analysis of 18F-FDG PET images in patients with extratemporal epilepsy and is, therefore, a useful tool in the evaluation of this patient group. The benefit of this analytical approach in patients with temporal epilepsy is less apparent.

Delayed rectifier potassium current in undiseased human ventricular myocytes.

OBJECTIVE: The purpose of the study was to investigate the properties of the delayed rectifier potassium current (IK) in myocytes isolated from undiseased human left ventricles. METHODS: The whole-cell configuration of the patch-clamp technique was applied in 28 left ventricular myocytes from 13 hearts at 35 degrees C. RESULTS: An E-4031 sensitive tail current identified the rapid component of IK (IKr) in the myocytes, but there was no evidence for an E-4031 insensitive slow component of IK (IKs). When nifedipine (5 microM) was used to block the inward calcium current (ICa), IKr activation was fast (tau = 31.0 +/- 7.4 ms, at +30 mV, n = 5) and deactivation kinetics were biexponential and relatively slow (tau 1 = 600.0 +/- 53.9 ms and tau 2 = 6792.2 +/- 875.7 ms, at -40 mV, n = 7). Application of CdCl2 (250 microM) to block ICa altered the voltage dependence of the IKr considerably, slowing its activation (tau = 657.1 +/- 109.1 ms, at +30 mV, n = 5) and accelerating its deactivation (tau = 104.0 +/- 18.5 ms, at -40 mV, n = 8). CONCLUSIONS: In undiseased human ventricle at 35 degrees C IKr exists having fast activation and slow deactivation kinetics; however, there was no evidence found for an expressed IKs. IKr probably plays an important role in the frequency dependent modulation of repolarization in undiseased human ventricle, and is a target for many Class III antiarrhythmic drugs.

Determinants of early and late results of combined valve operations and coronary artery bypass grafting.

BACKGROUND: Factors determining the outcome of operative correction of valvular abnormalities combined with coronary artery bypass grafting are still incompletely defined. METHODS: Determinants of early and late (more than 90 days) deaths and event-free survival were studied for combined valve operations and coronary artery bypass grafting in 741 patients using multivariate analysis. RESULTS: Ninety-day survival probability was 89% (95% confidence interval, 87% to 92%). Preoperative risk factors for early death were age, female sex, renal failure, New York Heart Association class IV or V, and mitral insufficiency. The operative risk factor was the duration of aortic cross-clamping. Five- and 10-year survival probabilities were 74% (95% confidence interval, 71% to 78%) and 43% (95% confidence interval, 36% to 50%), respectively. Preoperative risk factors for late death were age, preoperative renal failure, New York Heart Association class IV or V, vessel disease, and nonsinus rhythm. Five- and 10-year event-free survival probabilities were 57% (95% confidence interval, 53% to 61%) and 23% (95% confidence interval, 17% to 28%), respectively. Preoperative risk factors for non-event-free survival were age, female sex, reduced left ventricular function, mitral regurgitation, and pacemaker rhythm. CONCLUSION: The demographic factors of age and female sex; the comorbid condition of renal failure; the cardiac conditions of advanced New York Heart Association class, left ventricular function, mitral regurgitation, vessel disease, and cardiac rhythm; and the operative condition of ischemia time are the most important predictors of clinical outcome after combined valve operations and coronary artery bypass grafting.

Isolation of a full-length mitotic cyclin cDNA clone CycIIIMs from Medicago sativa: chromosomal mapping and expression.

Cyclins in association with the protein kinase p34cdc2 and related cyclin-dependent protein kinases (cdks) are key regulatory elements in controlling the cell division cycle. Here, we describe the identification and characterization of a full-length cDNA clone of alfalfa mitotic cyclin, termed CycIIIMs. Computer analysis of known plant cyclin gene sequences revealed that this cyclin belongs to the same structural group as the other known partial alfalfa cyclin sequences. Genetic segregation analysis based on DNA-DNA hybridization data showed that the CycIIIMs gene(s) locates in a single chromosomal region on linkage group 5 of the alfalfa genetic map between RFLP markers UO89A and CG13. The assignment of this cyclin to the mitotic cyclin class was based on its cDNA-derived sequence and its differential expression during G2/M cell cycle phase transition of a partially synchronized alfalfa cell culture. Sequence analysis indicated common motifs with both the A- and B-types of mitotic cyclins similarly to the newly described B3-type of animal cyclins.

Alfalfa Enod12 genes are differentially regulated during nodule development by Nod factors and Rhizobium invasion.

MsEnod12A and MsEnod12B are two early nodulin genes from alfalfa (Medicago sativa). Differential expression of these genes was demonstrated using a reverse transcription-polymerase chain reaction approach. MsEnod12A RNA was detected only in nodules and not in other plant tissues. In contrast, MsEnod12B transcripts were found in nodules and also at low levels in roots, flowers, stems, and leaves. MsEnod12B expression was enhanced in the root early after inoculation with the microsymbiont Rhizobium meliloti and after treatment with purified Nod factors, whereas MsEnod12A induction was detected only when developing nodules were visible. In situ hybridization showed that in nodules, MsEnod12 expression occurred in the infection zone. In empty Fix- nodules the MsEnod12A transcript level was much reduced, and in spontaneous nodules it was not detectable. These data indicate that MsEnod12B expression in roots is related to the action of Nod factors, whereas MsEnod12A expression is associated with the invasion process in nodules. Therefore, alfalfa possesses different mechanisms regulating MsEnod12A and MsEnod12B expression.

Mitral valve surgery combined with coronary bypass grafting: multivariate analysis of factors predicting early and late results.

Total of 123 patients (mean age: 63.8 +/- 7.3 years, (male/female 65/35 percent) underwent mitral valve surgery combined with coronary artery bypass grafting during a seven year period. Preoperatively 12% of them belonged to NYHA functional class II, 54% to class III, 29% to class IV and 3% was operated under emergency conditions. The mitral valve lesion was most frequently either ischaemic (45%) or rheumatic (33%) in origin. Left ventricle function was moderately decreased in 18% and severely damaged in 3% of the patients as documented by preoperative ventriculography. Coronary surgery was performed in all cases with an average number of distal anastomosis of 2.2 +/- 1.1 per patient. The hospital mortality was 13%. Risk factors for early and late mortality were determined by univariate and multivariate analysis. Advanced preoperative functional class and decreased left ventricular function or ischaemic etiology were identified as significant risk factors for early mortality. The patients were followed for an average of 33 +/- 25 months. The majority of them experienced significant functional improvement postoperatively with 69% belonging to NYHA class I or II. The late survival for the 107 hospital survivors was 94.7% at one year, and 84.7% at five years, respectively. Late survival was independently determined by preoperative functional class or previous myocardial infarction. Freedom from ischemic and valve related complications at five years was 95% and 71.2% respectively. 58.2% of the hospital survivors were in functional class I or II and free of any valve related or ischemic complications at the end of the fifth follow up year.(ABSTRACT TRUNCATED AT 250 WORDS)