Depression and recovery of reflex amplitude during electrical stimulation after spinal cord injury

ObjectiveThe aim of this study was to quantify, for the first time, H-reflexes evoked during prolonged trains of wide-pulse neuromuscular electrical stimulation (WP-NMES) in individuals with chronic spinal cord injury (SCI). We hypothesised that after the first H-reflex, reflex amplitudes would be depressed (due to post-activation depression), but would recover and this recovery would be enhanced after a “burst” of 100 Hz WP-NMES.MethodsSoleus M-waves and H-reflexes evoked during WP-NMES (1 ms pulse width) of the tibial nerve were quantified in nine individuals with SCI. WP-NMES was delivered in two patterns: “constant-frequency” (15 or 20 Hz for 12 s) and “burst-like” (15-100-15 Hz or 20-100-20 Hz; 4 s each phase) at an intensity that evoked an M-wave between 10% and 15% of the maximal M-wave (M_max).ResultsDuring constant frequency stimulation, after the initial depression from the first to the second H-reflex (1st: 57% M_max; 2nd: 25% M_max), H-reflexes did not recover significantly and were 37% M_max at the end of the stimulus train. During the burst-like pattern, after the initial depression (1st: 62% M_max; 2nd: 30%), reflexes recovered completely by the end of the stimulation (to 55% M_max) as they were not significantly different from the first H-reflex. M-waves were initially depressed (1st: 12% M_max; 2nd: 7% M_max) then did not change throughout the stimulation and were not significantly different between stimulation patterns. An analysis of covariance indicated that the depression in M-wave amplitude did not account for the depression in H-reflex amplitude.ConclusionsRelatively large H-reflexes were recorded during both patterns of NMES. The brief burst of 100 Hz stimulation restored H-reflexes to their initial amplitudes, effectively reversing the effects of post-activation depression.SignificanceFor individuals with chronic SCI, generating contractions through central pathways may help reduce muscle atrophy and produce contractions that are more fatigue-resistant for rehabilitation, exercise programs, or to perform activities of daily living.     

Optimal dye concentration and irradiance for laser-assisted vascular anastomosis

OBJECTIVE: This investigation was done in order to find optimal indocyanine green (ICG) concentration and energy irradiance in laser vascular welding. BACKGROUND DATA: Many studies have shown that laser tissue welding with albumin solder/ICG may be an effective technique in surgical reconstruction. However, there are few reports regarding optimal laser settings and concentrations of ICG within the albumin solder in laser-assisted vascular anastomosis. MATERIALS AND METHODS: Porcine carotid artery strips (n = 120) were welded in end-to-end by diode laser with 50% albumin solder of 0.01, 0.1, and 1.0 mM ICG at irradiance of 27.7, 56.7, and 76.9 W/cm(2), respectively. Temperature was measured by inserting thermocouples outside and inside the vessel. Tensile strength and histology were studied. RESULTS: Temperature and strength of the anastomosis significantly decreased (all p < 0.05) with increasing ICG concentration at 56.7 W/cm(2). Histological study showed minimal thermal injury limited to adventitia and no appreciable difference between all groups. CONCLUSIONS: ICG concentration within solder is the most important factor affecting both vascular temperature and tensile strength. The optimal balance between strength and minimal thermal injury may be achieved primarily at 56.7 W/cm(2) and 0.01 mM ICG.     

Role of raloxifene as a potent inhibitor of experimental postmenopausal polyarthritis and osteoporosis

OBJECTIVE: In postmenopausal rheumatoid arthritis (RA), both estrogen deficiency and the inflammatory disease contribute to the development of generalized osteoporosis. Hormone replacement therapy (HRT) with estradiol preserves bone mineral density (BMD) and ameliorates arthritis, but long-term therapy is no longer an option due to significant side effects. We therefore used a mouse model of human RA to test the hypothesis that a selective estrogen receptor modulator (SERM), the raloxifene analog LY117018, could be beneficial in the treatment of both arthritis and osteoporosis. METHODS: Female DBA/1 mice were ovariectomized and arthritis was induced with collagen immunization. Mice received an injection of raloxifene, estradiol, or vehicle control, administered prophylactically or therapeutically, and thereafter the clinical arthritis score was evaluated continuously. At termination, BMD was analyzed with peripheral quantitative computed tomography. Paws were collected for histology, and sera were analyzed for cytokines and markers of bone and cartilage turnover. Levels of cytokine messenger RNA (mRNA) were investigated with real-time polymerase chain reaction. RESULTS: Treatment with raloxifene dramatically decreased the frequency and severity of arthritis. Effective preservation of bone and cartilage was seen in raloxifene-exposed mice, as demonstrated by increased BMD and decreased serum levels of cartilage oligomeric matrix protein in the raloxifene-treated mice compared with controls. Decreased levels of mRNA for both tumor necrosis factor alpha and RANKL in spleen cells from raloxifene-treated arthritic mice indicated an immunosuppressive action of this SERM. CONCLUSION: In a well-established model of postmenopausal RA, the raloxifene analog LY117018 potently inhibits the progression of arthritis and the associated development of osteoporosis, both in a prophylactic and in a therapeutic regimen. Since long-term HRT has been associated with significant side effects, raloxifene may be a useful adjuvant treatment for postmenopausal RA.     

Changes in spinal but not cortical excitability following combined electrical stimulation of the tibial nerve and voluntary plantar-flexion

Unilateral training involving voluntary contractions, neuromuscular electrical stimulation (NMES), or a combination of the two can increase the excitability of neural circuits bilaterally within the CNS. Many rehabilitation programs are designed to promote such “neuroplasticity” to improve voluntary movement following CNS damage. While much is known about this type of activity-dependent plasticity for the muscles that dorsi-flex the ankle, similar information is not available for the plantar-flexors. Presently, we assessed the excitability of corticospinal (CS) and spinal circuits for both soleus (SOL) muscles before and after voluntary contractions of the right plantar-flexors (VOL; 5?s on–5?s off, 40?min), NMES of the right tibial nerve (tnNMES; 5?s on–5?s off, 40?min), or both together (V?+?tnNMES). CS excitability for the right (rSOL) and left SOL (lSOL) muscles was assessed by quantifying motor evoked potentials elicited by transcranial magnetic stimulation. Spinal excitability was assessed using measures from the ascending limb of the M-wave versus H-reflex recruitment curve. CS excitability did not change for rSOL (the activated muscle) or lSOL following any condition. In contrast, there was a marked increase in spinal excitability for rSOL, but only following V?+?tnNMES; the slope of the M-wave versus H-reflex recruitment curve increased?approximately twofold (pre?=?7.9; post?=?16.2) and H-reflexes collected when the M-wave was?~5?% of the maximal M-wave (M_max) increased by?~1.5×?(pre?=?19?% M_max, post?=?29?% M_max). Spinal excitability for lSOL did not change following any condition. Thus, only voluntary contractions that were coupled with NMES increased CNS excitability, and this occurred only in the ipsilateral spinal circuitry. These results are in marked contrast to previous studies showing NMES-induced changes in CS excitability for every other muscle studied and suggest that the mechanisms that regulate activity-dependent neuroplasticity are different for SOL than other muscles. Further, while rehabilitation strategies involving voluntary training and/or NMES of the plantar-flexors may be beneficial for producing movement and reducing atrophy, a single session of low-intensity NMES and voluntary training may not be effective for strengthening CS pathways to the SOL muscle.     

Long-term anti-arthritic and anti-osteoporotic effects of raloxifene in established experimental postmenopausal polyarthritis

Both oestrogen deficiency and the inflammatory disease contribute to the generalized bone loss seen in postmenopausal rheumatoid arthritis (RA). Oestradiol and the selective oestrogen receptor modulator raloxifene have been shown to ameliorate the disease in collagen-induced arthritis (CIA), a well-established animal model for human RA. The aim of this study was to investigate whether raloxifene-treatment would be beneficial in long-term treatment of established CIA, both regarding anti-arthritic and anti-osteoporotic properties. Female dilute brown agouti mice were ovariectomized and CIA was induced. Raloxifene or vehicle treatment was administered 5 days per week, and the clinical arthritis score was evaluated continuously. At termination, bone mineral density was analysed, paws were collected for histological examination and sera were analysed for markers of bone and cartilage turnover, as well as antibodies to type II collagen and levels of interleukin (IL)-6. Treatment with raloxifene is beneficial in long-term treatment of established CIA. It hampers the disease severity and frequency, protects the joints from destruction and protects against the development of osteoporosis. The proinflammatory cytokine IL-6 was down-regulated in raloxifene-treated mice compared with controls. The serum levels of antibodies to collagen were not affected by raloxifene-treatment. Long-term treatment with raloxifene has both anti-arthritic and anti-osteoporotic effects in established experimental postmenopausal polyarthritis.     

Effects of oestradiol and raloxifene on the induction and effector phases of experimental postmenopausal arthritis and secondary osteoporosis

Oestradiol and the selective oestrogen receptor modulator (SERM) raloxifene have been shown to ameliorate collagen-induced arthritis (CIA) in rats and in mice. One aim was to investigate if raloxifene exerts its anti-arthritic and anti-osteoporotic effects during the induction or effector phase of arthritis. A second aim was to analyse if raloxifene activates the oestrogen response element (ERE) to produce its immune-modulator effects. CIA or collagen-antibody-induced arthritis (CAIA) was induced in ovariectomized DBA/1-mice. CIA was used for evaluation of treatment during the induction, and CAIA for the effector phase of arthritis and osteoporosis development. Raloxifene, oestradiol or vehicle was administered 5 days/week. The clinical disease was evaluated continuously. Bone marrow density (BMD) was analysed with peripheral quantitative computer tomography, paws were collected for histological examination, and sera were analysed for markers of bone and cartilage turnover and proinflammatory cytokines. Transgenic luciferase (Luc)-ERE mice were immunized with collagen (CII), and after 10 days injected once with raloxifene, oestradiol or vehicle before termination. Spleens were analysed for luciferase activity to measure ERE activation. Treatment with oestradiol or raloxifene during the induction phase of CIA failed to affect arthritis. Raloxifene did not hamper disease activity in CAIA, whereas oestradiol delayed the onset and ameliorated the severity. Both raloxifene and oestradiol preserved BMD in CAIA. CII-immunization increased the oestradiol-induced ERE activation in spleen, and raloxifene activated the ERE at about 25% the intensity of oestradiol. Further experiments are needed to elucidate the exact mechanisms behind this finding.     

Elevated Aromatase Expression in Osteoblasts Leads to Increased Bone Mass Without Systemic Adverse Effects

The stimulatory effects of testosterone (T) on bone can either be through a direct activation of the androgen receptor (AR) or mediated through aromatization of T to estradiol (E2), followed by activation of estrogen receptors (ERs) in bone. Aromatase expression in osteoblasts and reproductive tissues is dependent on different promoters, which are differentially regulated. To study the effect of elevated local aromatization of T to E2 in bone, we developed a transgenic mouse model (Coll‐1α1‐Arom) that overexpresses the human aromatase gene under the control of the osteoblast specific rat type I α I procollagen promoter. The Coll‐1α1‐Arom mice expressed human aromatase mRNA specifically in bone and had unaffected serum E2 and T levels. Male Coll‐1α1‐Arom mice had clearly increased total body BMD, trabecular BMD, cortical BMD, and cortical thickness associated with elevated osteoprotegerin mRNA levels and reduced number of osteoclasts (p < 0.01). Treatment of ovariectomized mice with T increased cortical and trabecular thickness in the Coll‐1α1‐Arom mice (p < 0.001) but not in the wildtype mice. In conclusion, elevated aromatase expression specifically in osteoblasts results in stimulatory estrogenic effects in bone without increasing serum E2 levels. Because osteoblast‐specific aromatase expression results in an increased ER to AR activation ratio in bone, we propose that activation of ERs results in a more pronounced increase in bone mass than what is seen after activation of the AR. Development of osteoblast‐specific inducers of aromatase expression might identify substances with stimulatory effects on bone without systemic adverse effects.     

Amelioration of collagen‐induced arthritis and immune‐associated bone loss through signaling via estrogen receptor α, and not estrogen receptor β or G protein–coupled receptor 30

Objective

The effects of estrogen may be exerted via the nuclear estrogen receptors (ERs) ERα or ERβ or via the recently proposed transmembrane estrogen receptor G protein–coupled receptor 30 (GPR‐30). The purpose of this study was to elucidate the ER specificity for the ameliorating effects of estrogen on arthritis and bone loss in a model of postmenopausal rheumatoid arthritis (RA).

Methods

Female DBA/1 mice underwent ovariectomy or sham operation, and type II collagen–induced arthritis was induced. Mice were treated subcutaneously 5 days/week with the specific agonists propylpyrazoletriol (PPT; for ERα), diarylpropionitrile (DPN; for ERβ), G1 (for GPR‐30), or with a physiologic dose of estradiol. Clinical arthritis scores were determined continuously. At termination of the study, bone mineral density (BMD) was analyzed, paws were collected for histologic assessment, serum was analyzed for cytokines and markers of bone and cartilage turnover, and bone marrow was subjected to fluorescence‐activated cell sorting.

Results

Treatment with PPT as well as estradiol dramatically decreased the frequency and severity of arthritis. Furthermore, estradiol and PPT treatment resulted in preservation of bone and cartilage, as demonstrated by increased BMD and decreased serum levels of bone resorption markers and cartilage degradation markers, whereas no effect was seen after DPN or G1 treatment.

Conclusion

In a well‐established model of postmenopausal RA, ERα, but not ERβ or GPR‐30 signaling, was shown to ameliorate the disease and the associated development of osteoporosis. Since long‐term treatment with estrogen has been associated with significant side effects, increased knowledge about the mechanisms behind the beneficial effects of estrogen is useful in the search for novel treatments of postmenopausal RA.

     

GAD65 autoantibody epitopes in adult patients with latent autoimmune diabetes following GAD65 vaccination.

AIMS: Subcutaneous injection of recombinant human GAD65 (rhGAD65) in patients with latent autoimmune diabetes in adults (LADA) correlates with an increase in C-peptide levels. In this study we analysed the effect of rhGAD65 administration on the GAD65-specific autoimmune response. METHODS: Longitudinal serum samples obtained from LADA patients (n = 47) who received 4, 20, 100 or 500 microg alum-formulated rhGAD65 or placebo by subcutaneous injection twice (4 weeks apart) were analysed for their epitope recognition using GAD65-specific recombinant Fab and GAD65/67 fusion proteins. RESULTS: Overall, minor changes in the epitope pattern were observed using either approach. Only in the 500-microg dosage group was an increase in GAD65Ab level associated with a significant increase in the binding to a conformational epitope located at the middle part of GAD65. CONCLUSIONS: Our data suggest that the apparent beneficial effects of 20 microg alum-formulated recombinant human GAD65 is not explained by changes in the GAD65Ab epitope pattern.