Promotion of xenogeneic hematopoietic chimerism in rodents by mononuclear phagocyte depletion.

The successful establishment of tolerance toward pig tissues in primates through hematopoietic progenitor cell engraftment is restricted by the rapid disappearance of these cells in the recipient following infusion. We developed and tested the hypothesis that phagocytes of the reticuloendothelial system are responsible for the rapid clearance of infused pig hematopoietic cells using a mouse model. Mice received non-myeloablative conditioning and, on various days, were injected with medronate-encapsulated liposomes (M-L) or control blank liposomes, followed by the intravenous infusion of miniature swine hematopoietic cells. M-L were well-tolerated in mice (n=100) at levels that deplete mononuclear phagocytes. Depletion of mononuclear phagocytes in normal Balb/c mice as well as in severe combined immune deficient mice increased the accumulation of pig hematopoietic cells in the bone marrow (BM) by 10-fold when measured 24 h after the infusion of the cells. Colony-forming unit analysis showed an increased accumulation of pig hematopoietic progenitors in the BM of mice that were infused with medronate-liposomes. We conclude that depletion of mononuclear phagocytes by M-L has the potential to lower the barrier to the establishment of mixed chimerism and tolerance induction in xenotransplantation.

Gait characteristics of individuals with multiple sclerosis before and after a 6-month aerobic training program.

Individuals who have multiple sclerosis (MS) typically experience problems with physical activities such as walking, resulting from the combined effects of skeletal muscle weakness, sensory disturbances, spasticity, gait ataxia, and reduction in aerobic capacity. The aim of this study was to determine whether a 6-mo exercise program designed for aerobic conditioning might also affect gait abnormalities in individuals with MS. Subjects included 18 individuals with MS who presented a range of disability. Passive range of motion (PROM) in the lower limbs was measured and gait analyzed before and after exercise conditioning. Three-dimensional kinematics, ground reaction forces (GRF), and electromyographic information were acquired as subjects walked at self-selected velocities. Hip PROM increased following conditioning. Mean walking velocity, cadence, and posterior shear GRF (push-off force) decreased. During walking, maximum ankle dorsiflexion decreased and ankle plantarflexion increased. Total knee flexion/extension range during the walking cycle decreased slightly as did maximum hip extension. Results suggest this 6-mo training program had minimal effect on gait abnormalities.

Electrical stimulation-induced contraction to reduce blood stasis during arthroplasty.

Deep venous thrombosis and subsequent pulmonary embolism due to venous pooling/stasis commonly occur in patients during hip and/or knee arthroplasty (i.e., replacement). This problem may be alleviated by using techniques to promote lower limb blood flow. Electrical stimulation-induced contractions have been shown to activate the skeletal muscle pump, promote limb blood flow, and may be effective for reducing venous pooling/stasis and edema. Therefore, electrical stimulation may reduce the incidence of deep venous thrombosis (DVT) and pulmonary embolism (PE) during and following surgery. The overall goal of this project was to evaluate the clinical efficacy of sequential electrical stimulation-induced leg muscle contractions on the venous blood flow during surgery. The degree of venous pooling/stasis was monitored via electrical impedance changes in the thorax. The changes in the patient's central hemodynamics were then calculated. Thirty patients were recruited and randomly assigned to either a control group (n = 15, mean age = 66.4 +/- 7.3) or experimental group (n = 15, age = 60.7 +/- 9.7). Both groups received the standard medical treatment for prevention of DVT (i.e., coumadin, heparin, etc.) and compression stockings (TED, Kendall). The control group used the sequential compression device (SCD + TED) and the experimental group used electrical stimulation (ES + TED). Electrical stimulation was applied via surface electrodes to the lower-limb muscles (tibialis anterior and gastrocnemius) and upper limb muscles (quadriceps femoris and hamstrings). These muscles contracted sequentially, using an eight-channel electrical stimulator. Four seconds of calf (contraction/compression) were followed by 7-s of calf and thigh (contraction/compression) interspersed by 60-s rest period during both electrical stimulation or sequential compression device. This cycle continued throughout the surgery (60-75 min) for both groups. At 15 min intervals, venous return was monitored by impedance cardiograph. Physiologic responses including ventricular stroke volume (SV), cardiac output (CO), heart rate (HR), total peripheral resistance (TPR), as well as mean arterial pressure (MAP) were monitored. These responses were statistically analyzed and compared throughout the surgery within each group and between the two groups. The results show stroke volume and cardiac output to be higher throughout surgery in the electrical stimulation group as compared with the sequential compression device group. The heart rate was consistently lower during electrical stimulation for both groups. Total peripheral resistance did not change in the electrical stimulation group; but increased in the sequential compression device group. The data suggest that continuous electrical stimulation-induced contractions could improve lower leg circulation by eliciting the physiologic muscle pump. This will lead to improved venous circulation and reduction of blood stasis during total hip and/or knee surgery. This technique may offer greater protection against DVT and PE during surgery than the commonly used sequential compression device.

Specific tolerance across a discordant xenogeneic transplantation barrier.

Successful induction of tolerance across disparate (discordant) species barriers could overcome the organ shortage that presently limits clinical transplantation. We demonstrate here that xenogeneic swine thymic transplants can induce tolerance to swine antigens in mice, while positively selecting functional host CD4+ T cells. Immunologically normal C57BL/10 mice were thymectomized and depleted of T and natural killer cells; then they received transplants of fetal pig thymus and liver fragments. Mature mouse CD4+ T cells developed in the pig thymus grafts and migrated to the periphery. Swine grafts grew markedly and no anti-pig IgG response was produced. Mixed lymphocyte reactions confirmed that the new T cells were functional and were tolerant to pig antigens.

Functional neuromuscular stimulation. Exercise conditioning of spinal cord injured patients.

The results of numerous research studies indicate that functional neuromuscular stimulation (FNS)-induced exercise training of the paralyzed lower-limbs of spinal cord injured (SCI) patients can result in several important health and fitness benefits that cannot be achieved solely through conventional arm exercise modes. These benefits include peripheral adaptations where muscle strength, endurance, girth and appearance can be improved, as well as central circulatory adaptations which enhance aerobic (cardiopulmonary) capacity. There is also evidence that secondary medical complications related to sedentary lifestyle and inadequate circulation of blood can be prevented or alleviated with FNS exercise. The purpose of this paper is to present various FNS exercise modes including knee extension weight training, leg cycling and a hybrid exercise (voluntary-arm cranking and FNS-leg cycling), and to provide physiologic response patterns that are elicited.

The importance of nonimmune factors in reconstitution by discordant xenogeneic hematopoietic cells.

Bone marrow transplantation has been shown to induce donor-specific tolerance in rodent models. This approach could potentially be applied to xenotransplantation across discordant species barriers. To evaluate host factors resisting hematopoietic cell engraftment, we have developed two model systems utilizing the combination of swine into severe combined immunodeficient (SCID) mice. SCID mice lack functional B and T lymphocytes, and can therefore be used to evaluate nonimmune factors resisting marrow engraftment, and for adoptive transfer studies to test the role of immune cells and antibodies. First we transplanted swine bone marrow cells into SCID mice conditioned with whole-body irradiation (4 Gy). For nine weeks following the intravenous administration of 10(8) swine bone marrow cells, up to 3.8% of peripheral blood leukocytes were of swine origin, as determined by flow cytometry (FCM). These cells were all of the myeloid lineage. Swine IgG was also detectable in the serum for up to 14 weeks. The bone marrow of the reconstituted mice contained low percentages of swine myeloid cells, and swine myeloid progenitors could be detected for up to 20 weeks after bone marrow transplantation. In a second model, we grafted thymus and liver tissue from 45-69-day-old swine fetuses under the kidney capsule of 4 Gy-irradiated SCID mice. A suspension containing 10(8) swine fetal liver cells (FLC) was also administered i.p. Long-term repopulation with swine T cells was observed, with up to 1.5% swine T cells detected in the WBC, peritoneum, and spleen for at least 5.5 months postgrafting. These T cells expressed either CD4 or CD8, whereas up to 17.6% of cells in the thymic grafts expressed both CD4 and CD8. The i.p. FLC suspension was required for optimal long-term graft maintenance. Our studies show that (1) low level myeloid and B lymphocyte reconstitution can be achieved by transferring adult swine BMC to irradiated SCID recipients; (2) swine myeloid progenitors were detectable long-term in BMC of these mice, suggesting that stem cell engraftment was achieved; and (3) T cell reconstitution of SCID mice by swine progenitors requires cotransplantation of a swine stromal environment, as is provided by fetal swine thymus/liver grafts. We conclude that nonimmune factors such as those provided by species-specific stromal environments are important for reconstitution of some lineages by discordant hematopoietic stem cells.

Tibial bone density loss in spinal cord injured patients: effects of FES exercise.

A group of 37 spinal cord injured (SCI) patients underwent bone density measurements at the distal and proximal end of the tibia by a special computed tomography scanner, the OsteoQuant. Fifteen of these patients had follow-up measurements while enrolled in a lower-limb exercise training program with functional electrical stimulation (FES). The pre-exercise measurements revealed a strong correlation (0.88 < or = r < or = 0.90) of trabecular, subcortical, and cortical bone density between the distal and proximal ends of the tibia. The expected bone density loss during the first two years post injury (as calculated from the regression lines of bone density vs. time post injury) amounted to 51.5% for trabecular, 44.2% for subcortical, and 32.7% for cortical bone. No major bone density loss was calculated after 7 years post injury. Analysis of the bone density data during the FES exercise program revealed various degrees of loss. However, the rate of bone loss for this FES exercise group was less than expected from the regression lines. The reduction of bone loss was between 0.2 and 3.3% per year, and was significant (p < 0.05) for all bone parameters at the distal end and for trabecular bone density at the proximal end of the tibia. These bone density measurements revealed a potentially positive effect of FES exercise intervention for the rehabilitation of SCI patients.

The effects of functional electrical stimulation on shoulder subluxation, arm function recovery, and shoulder pain in hemiplegic stroke patients.

The purpose of this study was to evaluate the effectiveness of a functional electrical stimulation (FES) treatment program designed to prevent glenohumeral joint stretching and subsequent subluxation and shoulder pain in stroke patients. Twenty-six recent hemiplegic stroke patients with shoulder muscle flaccidity were randomly assigned to either a control group (n = 13; 5 female, and 8 male) or experimental group (n = 13; 6 female, and 7 male). Both groups received conventional physical therapy. The experimental group received additional FES therapy where two flaccid/paralyzed shoulder muscles (supraspinatus and posterior deltoid) were induced to contract repetitively up to 6 hours a day for 6 weeks. Duration of both the FES session and muscle contraction/relaxation ratio were progressively increased as performance improved. The experimental group showed significant improvements in arm function, electromyographic activity of the posterior deltoid, range of motion, and reduction in subluxation (as indicated by x-ray) compared with the control group. We concluded that the FES program was effective in reducing the severity of shoulder subluxation and pain, and possibly facilitating recovery of arm function.

Biomechanics of wheelchair propulsion during fatigue.

Musculoskeletal injuries can result from overuse or incorrect use of manual wheelchairs, and can hinder rehabilitation efforts. The purpose of this study was to investigate wheelchair propulsion biomechanics of spinal cord injured, non-athletic wheelchair users. We studied changes in the variables that occur with fatiguing wheelchair propulsion to determine how the user's physical characteristics and the state of fatigue influence risk of injury. Twenty male paraplegic patients were videotaped during propulsion to fatigue on a stationary, instrumented wheelchair positioned on a roller with adjustable frictional resistance. Peak handrim force was significantly correlated with concentric shoulder flexion and elbow extension isokinetic torques. Significant changes (p < 0.05) with fatigue were found in increased peak handrim force, decreased ulnar/radial deviation range of motion, and increased trunk forward lean. Of the three upper extremity joints, highest calculated joint moments were found in shoulder flexion (p < 0.05). These biomechanical results suggest that potentially harmful changes occur with fatigue, and that the shoulder may be the most prone to musculotendinous-type overuse injury.

Evaluation of Muscle Performance and Cardiopulmonary Fitness in Patients with Multiple SclerosisImplications for Rehabilitation.

Multiple sclerosis (MS) is a neurological disease characterized by a variety of potentially debilitating symptoms. The manner in which the disease affects each individual is unique. Many individuals with MS have minimal neuromuscular impairment and remain ambulatory throughout their lives, whereas others experience more rapid disease progression and loss of functional independence. Although appropriate exercise testing and training could potentially improve clinical treatment and rehabilitation outcome, very little research has focused on understanding how MS affects basic physiologic responses during exercise. Objective measurements of muscle function and cardiopulmonary (aerobic) fitness have indicated variable results depending upon the disease severity. Patient-specific information is important for establishing realistic guidelines for therapeutic exercise programs. It appears that appropriate training can improve both skeletal muscle function and cardiopulmonary fitness. However, current clinical indices of neurologic function do not consider physical capability as part of the criteria. Thus, it seems desirable to supplement these criteria with objective tests of exercise performance. Special precautions and considerations for exercise testing of individuals with MS need to be incorporated to ensure safety and accurate interpretation of data.

Functional electrical stimulation leg cycle ergometer exercise: training effects on cardiorespiratory responses of spinal cord injured subjects at rest and during submaximal exercise.

This study investigated the cardiorespiratory (CR) responses at rest and during submaximal (0-W) functional electrical stimulation (FES)-induced leg cycle ergometer (LCE) exercise prior to and following a progressive intensity FES-LCEa exercise training program in spinal cord injured (SCI) subjects. Seven quadriplegics and six paraplegics participated in FES-LCE training three sessions per week for approximately 12 weeks (36 sessions). Monitored CR responses, including oxygen uptake (VO2), pulmonary ventilation (VE), respiratory exchange ratio (RER), arteriovenous O2 difference (a-vO2), blood pressure (BP), heart rate (HR), stroke volume (SV), total peripheral resistance (TPR), and cardiac output (Q), were determined before and after training. Power output (PO) increased significantly (p < .05) over the duration of the training program, indicating increased in strength and endurance of the paralyzed muscles used. Respiratory responses were not significantly altered by training in both groups. FES-LCE training significantly increased resting HR and SBP in quadriplegics and lowered SBP, DBP, and MAP in paraplegics. In both groups, HR and BP during submaximal exercise significantly decreased and SV and Q significantly increased after completion of the training program. These results suggest that FES-LCE training improves peripheral muscular and central cardiovascular fitness in SCI subjects. Posttraining HR and BP may also be more stable in quadriplegics and alleviate hypotension. This therapeutic exercise may ultimately lead to improved rehabilitation outcome and reduced stress during activities of daily living, and possibly reduce the risks for secondary CR disabilities.

Persistence of lpr/lpr-derived IgG2a secreting B cells in normal----lpr/lpr adult radiation chimeras or lpr/lpr----normal kidney capsule chimeras.

Lethally irradiated MRL/lpr mice reconstituted with bone marrow stem cells from a normal mouse strain develop a state of split hematopoietic chimerism; erythrocytes, granulocytes, and macrophages are derived from the normal stem cell inoculum while the peripheral T lymphocytes are derived from radioresistant lpr host cells. Moreover, these mice have normal levels of serum IgM and IgG2a produced by radioresistant host B cells, even though they have relatively few sIgM+ B cells. In order to better understand the differentiation and regulation of B cells present in these chimeric mice, the current study was undertaken to localize and to assess the functional capacity of the lpr B cells producing the serum antibodies. Surface IgG2a+ cells could not be found in the spleen or lymph nodes of these mice, but large lymphocytes containing cytoplasmic IgG2 of host (lpr) allotype could be readily detected, even though they constituted less than 1% of the total spleen population. The host-derived serum IgG2 and IgG2+ cells were even present in the spleens of "leaky" mice that had relatively normal numbers of donor-derived sIgM+ B cells. These lpr B cells secreted IgG2a antibody that bound ssDNA, but they could not respond to immunization with SRBC. These results indicate that the lpr-derived radioresistant B cells have a limited capacity for proliferation and are already committed to the memory lineage. The presence of similar B cells in normal mice transplanted with neonatal lpr/lpr spleen fragments suggests that lpr/lpr B cell development is inherently abnormal.

Physiologic effects of electrical stimulation leg cycle exercise training in spinal cord injured persons.

The purpose of this study was to assess the physiologic training effects of functional electrical stimulation leg cycle ergometer (FES-LCE) exercise in persons with spinal cord injury (SCI) who were previously untrained in this activity. Ten persons with quadriplegia (C5 to C7) and eight with paraplegia (T4 to T11) performed FES-LCE training on an ERGYS I ergometer 10 to 30 minutes per day, 2 or 3 days per week for 12 to 16 weeks (36 total sessions). Training session power output (PO) ranged from 0.0W (no external resistance) to 30.6W. Each subject completed discontinuous graded FES-LCE and arm crank ergometer (ACE) tests before and after training for determinations of peak lower and upper extremity metabolic, pulmonary, and hemodynamic responses. Compared with pretraining, this SCI group exhibited significantly (p less than or equal to .05) higher posttraining peak PO (+45%), oxygen uptake ([O2], + 23%), pulmonary ventilation (+27%), heart rate (+11%), cardiac output ([Qt], + 13%) and significantly lower total peripheral resistance ([TPR], - 14%) during FES-LCE posttests. There were no significant changes in peak stroke volume (+6%), mean arterial pressure ([MAP], - 5%), or arteriovenous oxygen difference ([a-vO2diff], + 10%) during posttraining FES-LCE tests. In addition, no significant differences were noted for the peak level of any monitored variable during ACE posttests after FES-LCE training. The rise in total vascular conductance, implied by the significant decrease in posttraining TPR during FES-LCE tests, denotes that a peripheral circulatory adaptation developed in the persons with SCI during FES-LCE exercise training.(ABSTRACT TRUNCATED AT 250 WORDS)

Concentric and eccentric isokinetic lower extremity strength in persons with multiple sclerosis.

This study was supported by the Levine-Rubenstein MS Research Fund and The Rehabilitation Research and Development Service of the U.S. Department of Veterans Affairs. Clinicians might be reluctant to institute exercise training programs for individuals with multiple sclerosis (MS) because so little information is available regarding their muscle performance capability. The purpose of this project was to compare the quadriceps and hamstrings muscle groups' torque capacity of individuals with MS (N = 9) to matched non-MS controls (N = 9). Muscle torques at several speeds were measured using a KIN-COM II isokinetic dynamometer during both concentric (30, 60, and 90 degrees /sec) and eccentric (45, 60, and 75 degrees /sec) contractions. Peak torque for both muscle groups at all velocities were higher for the non-MS group; however, this difference was only significant for the concentric quadriceps muscle contraction (p < .05). All subjects demonstrated higher torque eccentrically than concentrically for the quadriceps, but these differences were only significant for the experimental subjects. The MS group showed a greater decrease in torque with increasing speed than the non-MS group. Deficits between the MS and control groups related to changes in contraction velocity were greater during concentric than eccentric contractions. These findings suggest that strengthening programs that emphasize concentric exercise at the higher of the three speeds of movement included in this study may be most effective in treating this particular type of strength deficit in MS patients. J Orthop Sports Phys Ther 1992;16(3):114-122.

Metabolic and hemodynamic responses to concurrent voluntary arm crank and electrical stimulation leg cycle exercise in quadriplegics.

This study determined the metabolic and hemodynamic responses in eight spinal cord injured (SCI) quadriplegics (C5-C8/T1) performing subpeak arm crank exercise (ACE) alone, subpeak functional electrical stimulation leg cycle exercise (FES-LCE) alone, and subpeak FES-LCE concurrent with subpeak ACE (hybrid exercise). Subjects completed 10 minutes of each exercise mode during which steady-state oxygen uptake (VO2), pulmonary ventilation (VE), heart rate (HR), cardiac output (CO), stroke volume (SV), mean arterial pressure (MAP), arteriovenous oxygen difference (a-v O2 diff), and total peripheral resistance (TPR) were determined. Although mean VO2 for both ACE alone and FES-LCE alone was matched at 0.66 l/mi, individualized power outputs ranged from 0-30 W (mean = 19.4 +/- 1.3) and 0-12.2 W (mean = 2.3 +/- 0.6), respectively. Hybrid exercise elicited significantly higher VO2 (by 54 percent), VE (by 39-53 percent), HR (by 19-33 percent), and CO (by 33-47 percent), and significantly lower TPR (by 21-34 percent) than ACE or FES-LCE performed alone (P less than or equal to 0.05). Stroke volume was similar between hybrid exercise and FES-LCE alone, and these two exercise modes evoked a significantly higher SV (by 41-56 percent) than during ACE alone. These data clearly demonstrate that hybrid exercise creates a higher aerobic metabolic demand and cardiac-volume load in SCI quadriplegics than either subpeak levels of ACE or FES-LCE performed separately. Therefore, hybrid exercise may provide more advantageous central cardiovascular training effects in quadriplegics than either ACE or FES-LCE alone.

Adaptive control of functional neuromuscular stimulation-induced knee extension exercise.

An automated system for exercising the paralyzed quadriceps muscles of spinal cord injured patients using functional neuromuscular stimulation (FNS) has been developed. It induces smooth concentric and eccentric contractions in both limbs to enable bilateral 70 degree knee extensions in an asynchronous pattern. External load resistance is applied at the ankle level to "overload" the muscles and bring about training effects. The system uses adaptive control methods to adjust FNS current output (threshold level and the ramp slope) to the quadriceps muscles to maintain performance as the muscles fatigue. Feedback control signals for limb movement and knee extension angle are used to continuously adjust the FNS current parameters so that the external load is moved through the preset zero to 70 degree angle range. Typically, the threshold current level and the FNS current increase as the muscles fatigue to maintain performance with repetitive contractions. Fatigue is defined as the inability to extend the knee to 50 percent of the 70 degree target angle. When this occurs, FNS is automatically terminated for the fatigued leg, while the functioning leg continues to exercise. The automated nature of this system appears to be advantageous as compared to a manually operated system for subject safety, convenience, and uniformity of exercise bouts. Simulated safety problems, such as hyperextension of the knee joint, open circuitry, muscle spasms, and low battery power, were successfully detected by the logic circuitry, and the system followed appropriate safety procedures to minimize risk.

Musculoskeletal responses of spinal cord injured individuals to functional neuromuscular stimulation-induced knee extension exercise training.

This study was conducted to evaluate a newly designed functional neuromuscular stimulation (FNS)-induced knee extension (KE) exercise system that incorporates the most desired features of previously described systems by determining the musculoskeletal responses of spinal cord injured (SCI) individuals to training. A specially designed chair and electrical stimulator were fabricated for FNS-induced KE resistance exercise. Surface electrodes were placed over motor points of the quadriceps muscles, and KE was alternated between legs at an average rate of 6 KE/min/leg. KE testing protocols were developed for pre- and post-training evaluations of performance, and 12 SCI subjects exercise-trained up to three times per week for 36 sessions using a progressive resistance load at ankle level. Pre- and post-training evaluation data were statistically compared using a 0.05 level for significance. Quadriceps muscle performance (strength x repetitions) improved for both legs in all subjects as indicated by significant increases in load resistance and repetitions over the 36-session training period (right leg mean = 1156.0 versus 1624.8 kg.reps, left leg mean = 1127.3 versus 1721.1 kg.reps). In addition, knee range of motion significantly increased (right leg mean = 134 versus 146 degrees, left leg mean = 133 versus 144 degrees). Thigh skinfold, thigh girth, body weight and bone density were not significantly changed. The lack of decrease in bone density in some subjects suggests that the training may retard the rate of bone loss which typically occurs with SCI. No injuries or problems were encountered during testing and training.(ABSTRACT TRUNCATED AT 250 WORDS)