Long-term results of the anterior floating method for cervical myelopathy caused by ossification of the posterior longitudinal ligament.

STUDY DESIGN: Results of the anterior floating method used to decompress ossification of the posterior longitudinal ligament were studied for an average postoperative interval of 13 years. OBJECTIVE: To investigate the long-term results of the anterior floating method used to manage ossification of the posterior longitudinal ligament. SUMMARY OF BACKGROUND DATA: The anterior floating method is a technique that differs from the extirpation method used to manage ossification of the posterior longitudinal ligament. Reports of the long-term results from anterior decompression used to manage cervical ossification of the posterior longitudinal ligament are rare. METHODS: The anterior floating method was used to decompress cervical ossification of the posterior longitudinal ligament in 63 patients. These patients were followed for more than 10 years with neurologic evaluations using a scoring system proposed by the Japanese Orthopedic Association (JOA score). RESULTS: The recovery rate was 66.5% at 10 years and 59.3% at 13 years, the time of the final survey. Operative outcomes most closely reflected the preoperative duration and severity of myelopathy (JOA score) and the preoperative cross-sectional area of the spinal cord. There was no correlation with the canal narrowing ratio or the thickness of ossification of the posterior longitudinal ligament. Delayed deterioration was attributed to an original inadequate decompression and progression of ossification of the posterior longitudinal ligament outside the original operative field. There was no evidence of significant recurrent ossification of the posterior longitudinal ligament within the margins of prior decompression. CONCLUSIONS: The anterior floating method appears to yield adequate long-term outcomes when used to manage ossification of the posterior longitudinal ligament.

Endovascular treatment of ruptured dissecting aneurysms aimed at occlusion of the dissected site by using Guglielmi detachable coils.

OBJECT: Surgical or endovascular occlusion of the parent artery proximal to an aneurysm has been recommended for treatment of dissecting aneurysms of the intracranial posterior circulation. However, dissecting aneurysms may rupture even after proximal occlusion because distal progression of thrombus is necessary to occlude the dissecting aneurysm completely, and this may be delayed by the presence of retrograde flow. In this article the authors present their experience in treating six patients with ruptured dissecting aneurysms. METHODS: The authors report on six patients with a ruptured dissecting aneurysm in the posterior fossa who were successfully treated by endovascular occlusion of the aneurysm by using Guglielmi detachable coils. The procedure was particularly aimed at occluding the dissected site. CONCLUSIONS: At the present time, endovascular occlusion of the dissected site is a safe, minimally invasive, and reliable treatment for dissecting aneurysms when a test occlusion is tolerated and adequate collateral circulation is present.

Calpain inhibitor entrapped in liposome rescues ischemic neuronal damage.

Transient forebrain ischemia induces activation of calpain and proteolysis of a neuronal cytoskeleton, fodrin, in gerbil hippocampus. This phenomenon precedes delayed neuronal death in hippocampal CA1 neurons. We examined effects of a calpain inhibitor on delayed neuronal death after transient forebrain ischemia. In gerbils, a selective calpain inhibitor entrapped in liposome was given transvenously and 30 min later, 5-min forebrain ischemia was produced by occlusion of both common carotid arteries. On day 7, CA1 neuronal damage was examined in the hippocampal slices stained with cresyl violet. Calpain-induced proteolysis of fodrin was also examined by immunohistochemistry and immunoblot. Additionally, to assure entrapment of the inhibitor by CA1 neurons, the inhibitor-liposome complex was labeled with FITC and given to gerbils. Fluorescence in the hippocampal slices was examined by confocal laser scanning microscope. Selective CA1 neuronal damage induced by forebrain ischemia was prevented by administration of the inhibitor in a dose-dependent manner. Calpain-induced proteolysis of fodrin was also extinguished by the calpain inhibitor in a dose-dependent manner. Bright fluorescence of the FITC-labeled inhibitor was observed in the CA1 neurons. The data show an important role of calpain in the development of the ischemic delayed neuronal death. Calpain seems to produce neuronal damage by degrading neuronal cytoskeleton. Our data also show a palliative effect of the calpain inhibitor on the neurotoxic damage, which offers a new and potent treatment of transient forebrain cerebral ischemia.

Anterior floating method for cervical myelopathy caused by ossification of the posterior longitudinal ligament.

Ossification of the posterior longitudinal ligament lessens the sagittal diameter of the cervical canal and compresses the spinal cord anteriorly, and may produce severe disabling myelopathy. The anterior floating method is one of the anterior decompression and reconstructions used in the treatment of cervical myelopathy caused by ossification of the posterior longitudinal ligament. This procedure consists of subtotal resection of vertebral bodies and discs, with slight thinning and release of the ossified ligament using air instrumentation. This is followed by reconstruction of the cervical spine using autogenous strut bone graft accompanied by postoperative application of a halo vest. This method is indicated for patients who present with moderate or severe myelopathies, and especially in those where the canal narrowing ratio exceeds 60%. This radical procedure causes decompression of the spinal cord and restores its function by enlarging the neural canal with anterior migration of the ossified ligament. The procedure minimizes the extent of surgical invasions and avoids damage to the neural tissue, because it does not require the removal of the ossification of the posterior longitudinal ligament. It also stops postoperative regrowth of the ossification. The operative results with long term followup indicate a 71% average recovery rate based on the criteria established by the Japan Orthopedic Association.

Activation of protein kinases in canine basilar artery in vasospasm.

Subarachnoid hemorrhage (SAH) often leads to a long-term narrowing of cerebra! artery called vasospasm. To understand the molecular mechanisms in vasospasm, signal transduction of tyrosine kinase pathway and phosphorylation of myosin light chain (MLC) and calponin (CaP) in the basilar artery were studied. Vasospasm was produced in the canine basilar artery by a two-hemorrhage method, and vasocontraction was induced by a local application of KCI or serotonin to the basilar artery after a transclival exposure. Intracellular substrates of tyrosine kinase pathway, including Shc, Rafl, and extracellular-regulated kinases in the basilar artery, were activated after SAH, and the activation of Shc suggests stimulation of signal transductions from tyrosine kinase receptors, G-coupled receptors, or both. The activation of tyrosine kinase pathway in vasospasm also was supported by dose-dependent dilation of the spastic basilar artery on days 0 and 7 by topical application of genistein, a tyrosine kinase inhibitor, and associated marked inhibition of tyrosine phosphorylation of intracellular substrates, including Shc. In addition, the generation of protein kinase M, catalytic fragment of protein kinase C(alpha) (PKC alpha), in vasospasm on days 0 and 7 was inhibited in response to genistein, indicating an inactivation of mu-calpain. It is suggested, therefore, that the reversal of vasospasm by genistein is closely associated with the restoration of intracellular Ca2+ levels. However, the increased activities of Raf1 and extracellular-regulated kinases in vasospasm were declined on day 7 compared with those on day 0 or 2, suggesting that the activation of tyrosine kinase pathway is more closely associated with the early stage of vasospasm than with the late stage of vasospasm. The analysis by pyrophosphate polyacrylamide gel electrophoresis (PPi-PAGE) demonstrated three MLC bands in vasospasm on days 2 and 7, as well as in KCI- and serotonin-induced vasocontraction. Since PPi-PAGE resolves smooth muscle MLC into three bands in the MLC kinase (MLCK)-mediated phosphorylation and into a single band in the PKC-mediated phosphorylation based on the phosphorylation state, the current results suggest that MLC in vasospasm is phosphorylated by MLCK but not by PKC. In basilar artery, CaP was significantly down-regulated, and in addition, significantly phosphorylated on serine and threonine residues only in vasospasm on days 2 and 7. Although the significance of CaP phosphorylations in vivo still is controversial, CaP down-regulation and phosphorylation may attenuate the inhibition of Mg(2+)-ATPase activity by CaP and induce a potential enhancement of smooth muscle contractility in delayed vasospasm. Since CaP is phosphorylated in vivo by PKC, activated PKC in vasospasm may phosphorylate CaP. Thus, SAH stimulates tyrosine kinase pathway to increase intracellular Ca2+ and activate PKC, and the former activates MLCK to phosphorylate MLC, whereas the latter phosphorylates CaP but not MLC.

Spinal cord herniation into an extensive extradural meningeal cyst: postoperative analysis of intracystic flow by phase-contrast cine MRI.

We report a patient with idiopathic spinal cord herniation who underwent postoperative myelography and phase-contrast cine magnetic resonance imaging (MRI) to clarify the condition of the ventral cystic lesion into which the spinal cord had been herniated. This 58-year-old man showed symptoms and signs compatible with Brown-Séquard syndrome. A myelogram and MRI showed that the spinal cord was incarcerated in a small space, and this finding led to the diagnosis of spinal cord herniation. In the operation, the herniated spinal cord was returned to the initial intradural space, and the dural defect was enlarged to prevent recurrence of the herniation. After the operation, leg muscle strength improved and bladder and bowel dysfunction resolved, but analgesia was unchanged. Postoperative myelography revealed a very large extradural cystic lesion, which extended from C2 to L1 vertebral level. A cerebrospinal fluid (CSF) flow study of the intradural space disclosed a near-normal pattern, but the intracystic CSF dynamics were abnormal. In the intracystic space at just the upper level of the lesion, cranial flow (reverse directional flow in comparison with the intradural flow) was seen in the systolic cycle after momentary fast caudal flow, and these CSF dynamics may have been related to the spinal cord herniation.

Biological and structural properties of cyclic peptides derived from the alpha-amylase inhibitor tendamistat.

Six cyclic peptides with 5, 7, 9, 11, 13, and 15 amino acids, with the inhibitory sequence of the alpha-amylase inhibitor tendamistat, were synthesized. The 11-residue peptide inhibited porcine pancreatic alpha-amylase most potently (K1 0.29 +/- 0.09 microM). For the 11-residue peptide, the circular dichroism study suggested a preliminary relationship between its inhibitory activity and structural property.

Purification and some properties of endo-1,4-beta-D-xylanase from a fresh-water mollusc, Pomacea insularus (de Ordigny).

Endo-1,4-beta-D-xylanase (EC 3.2.1.8) was purified from viscera of a fresh-water mollusc, Pomacea insularus (de Ordigny). The purified enzyme, with a molecular weight of 47,000, gave a single protein band in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The amino-terminal sequence was Ala-Ala-Gly-Ala-Gly-Val-Thr-Ser-Glu-Lys-Asp-Arg-Leu-Arg-Arg-Ser-Asp-Lys -Thr-Val-His-Val-Asn-. The enzyme was stable from pH about 4.5 to 9.5 and had its maximum activity at pH about 5.5. The purified enzyme produced X2, X3, X4, and larger xylooligosaccharides from birchwood xylan. The enzyme activity was greatly inhibited by Ag+, Hg2+, Cu2+, N-bromosuccinimide, and p-chloromercuribenzoic acid. On the other hand, the enzyme activity was greatly elevated by the addition of chloride ion.

Lumbar nerve root compression caused by lumbar intraspinal gas. Report of three cases.

STUDY DESIGN: Three cases of lumbar nerve root compression associated with intradiscal gas formation and its migration are reported. OBJECTIVES: To establish the pathogenic mechanism by which gas is formed in the spinal canal. SUMMARY OF BACKGROUND DATA: Few such cases have been reported of patients with clinical symptoms of lumbar radiculopathy resulting from gas in the spinal canal. METHODS: Surgery with needle aspiration of the gas was performed in all cases. RESULTS: The presence of the gas was detected in the extradural space in one patient and within the nerve sheath in two patients. After surgery, all patients recovered well, and there were no obvious remnant neurologic abnormalities. One year later, all patients were completely free of symptoms. CONCLUSIONS: Surgery with needle aspiration of the gas resulted in clinical improvement in all cases, confirming that intraspinal gas is an important cause of lumbar radiculopathy. The composition of the gas in one patient was analyzed by gas chromatography, which revealed an overwhelming preponderance of nitrogen. A relationship between a gas-containing pseudocyst in the spinal canal and a degenerated intervertebral disc was identified, a key finding for understanding the precise nature of this disorder. Intradiscal gas formation and its outward migration as a sequel of intervertebral disc degeneration also has been addressed in this report.

Cytotoxic fragment of amyloid precursor protein accumulates in hippocampus after global forebrain ischemia.

We developed an antibody specific to beta-amyloid precursor protein (beta APP) fragments possessing the exact amino terminus of the beta-amyloid peptide and examined its induction in postischemic hippocampus. In control hippocampus, this APP fragment was lightly observed in pyramidal neurons of CA sectors and dentate granule cells. Transient forebrain ischemia enhanced accumulation of the APP fragment in CA1 pyramidal neurons. Seven days after the ischemia, while the APP fragment was still observed in dentate granule cells and CA3 neurons, it disappeared in dead CA1 neurons. While astrocytes did not show in any immunoreactivity throughout the experiment, those in the CA1 sector showed moderate immunoreactivity 7 days after the ischemia. The APP fragment has a cytotoxic effect on cultured neurons. These results suggest that the accumulation of the cytotoxic APP fragment in CA1 neurons may play a role in the development of delayed neuronal death after the ischemic insult.

Comparison of thermal and guanidine hydrochloride denaturation behaviors of glucoamylase from the STA1 gene of Saccharomyces cerevisiae var. diastaticus.

To investigate the relationships between enzyme inactivation and conformational change, the effects of heat and guanidine hydrochloride (GnHC1) on the STA1 gene glucoamylase (STA1GA) of Saccharomyces cerevisiae var. diastaticus were examined by circular dichroism and fluorescence spectroscopies. A conformational change was observed in the thermal denaturation of STA1GA, while extensive enzyme inactivation occurred in GnHC1 denaturation before noticeable conformational change.

Selecting a surgical method for thoracic myelopathy caused by ossification of the posterior longitudinal ligament.

STUDY DESIGN: The authors classified typical distributional patterns of ossification of the posterior longitudinal ligament of the thoracic spine in 1) central part of S-curve, 2) just above apical vertebra, and 3) combined with ossification of ligamentum flavum below apical vertebra. The results of the surgical methods selected according to the authors' classification were compared with those of previous reports. OBJECTIVES: To establish the criteria for selecting an appropriate surgical method for ossification of the posterior longitudinal ligament of the thoracic spine. SUMMARY OF BACKGROUND DATA: Poor surgical results for ossification of the posterior longitudinal ligament of "middle or lower thoracic spine" have been reported, but the unsuccessful location and curve has not been strictly defined. METHODS: The authors studied postsurgical results in 26 cases of thoracic myelopathy caused by ossification of the posterior longitudinal ligament. They also investigated radiographs of 111 cases, including 85 patients under observation, and examined the relationships between thoracic spine alignment and ossification of the posterior longitudinal ligament distribution. RESULTS: Twenty-three patients treated with methods conforming to the authors' criteria achieved satisfactory recovery in walking ability except for one patient. The results of the other three patients who underwent surgery with nonconforming methods were uneven. CONCLUSION: Posterior decompression, as well as anterior decompression, is effective in the first pattern in the cervicothoracic region. In case of the second pattern, the responsible ossification of the posterior longitudinal ligament always lies one or two levels above the apical vertebra and should be removed by anterior approach, regardless of the extent of kyphosis. Transthoracic anterior decompression surgery is considered the best method for most patients under the second and third patterns.

Purification and some properties of an endo-1,4-beta-D-mannanase from a marine mollusc, Littorina brevicula.

An endo-1,4-beta-D-mannanase (EC 3.2.1.78) was purified from viscera of a marine mollusc Littorina brevicula. The purified enzyme, with a molecular weight of 42,000, was homogeneous by SDS-PAGE. The amino-terminal sequence starting with Gly was analyzed up to the 30th amino acid. The enzyme was stable from pH about 4.0 to about 9.0 and had its maximum activity at pH about 6.5. The purified enzyme produced M2, M3, M4, and M5 from Codium beta-1,4-mannan. The enzyme activity was greatly inhibited by Ag+, HG2+, Cu2+, and N-bromosuccinimide at 1 mM concentration.

The natural history of herniated nucleus pulposus with radiculopathy.

STUDY DESIGN: The present study retrospectively investigated the morphologic changes that occurred during conservative treatment of patients with unilateral leg pain resulting from herniated nucleus pulposus without significant lumbar canal stenosis. OBJECTIVES: The results were correlated with clinical outcomes and extruding forms to determine which type of herniated nucleus pulposus had the greatest capacity for spontaneous regression and how rapidly such regression might occur. SUMMARY OF BACKGROUND DATA: The study population consisted of 77 patients with radiculopathy. METHODS: All patients complained primarily of unilateral leg pain, and 94% had positive tension signs. Additionally, 32% exhibited muscle weakness corresponding to the symptomatic nerve root. All patients were studied more than twice using magnetic resonance imaging during conservative therapy at a mean interval of 150 days. Morphologic changes on magnetic resonance imaging fell into four categories, with herniated nucleus pulposus classified into three types using T1-weighted sagittal views. Each patient was reexamined on the same scanner; 53 patients were examined twice, and 24 patients were examined more than three times. RESULTS: Morphologic changes, with the exception of 13 false-negative cases, basically corresponded to clinical outcome. In half of the cases that showed some improvement at follow-up evaluation, improvement of clinical findings were seen before those observed on magnetic resonance imaging. Migrating herniated nucleus pulposus frequently presented an obvious decrease in size, and even disappearance in seven cases. The further the herniated nucleus pulposus migrated, the more decrease in size could be observed. The cases apparently corresponding to "protrusion" showed little or no change on follow-up magnetic resonance imaging. Regarding the mechanism of herniated nucleus pulposus disappearance, exposure to the vascular supply undoubtedly took a part, although many factors were suspected to have some influence. CONCLUSION: Morphologic changes on magnetic resonance imaging mainly corresponded to clinical outcomes but tended to lag behind improvement of leg pain. Disappearance of herniate nucleus pulposus was seen frequently in the cases of migrating disc herniation, and it was presumed that exposure to the vascular supply had a lot to do with this phenomenon.

Improved Purification and Spectroscopic Properties of Squash Glutamate Decarboxylase.

Squash glutamate decarboxylase was purified by DEAE-Cellulose batchwise followed by Blue-Sepharose, Cellulofine GCL-2000, and Toyopearl HW-55F column chromatography. The purified glutamate decarboxylase had a high specific activity (95.0 u/mg). The absorption spectrum of glutamate decarboxylase had an absorption maximum at 420 nm in the range 300-500 nm. A pH change from 5.3 to 7.8 was accompanied by a decrease in absorbancy at 420 nm. One mole of glutamate decarboxylase contained 3.8 and 1.3 mol of pyridoxal 5'-phosphate at pH 5.8 and pH 7.8, respectively.

Activity of smooth muscle phosphatases 1 and 2A in rabbit basilar artery in vasospasm.

BACKGROUND AND PURPOSE: Subarachnoid hemorrhage frequently leads to a long-term cerebral artery narrowing called vasospasm. Recently, the involvement of myosin light chain kinase has been found in experimental vasospasm in our laboratory. We therefore measured the activity of serine/threonine protein phosphatases 1 and 2A in the rabbit basilar artery in vasospasm and in vasocontraction to study their role, particularly in regard to vasospasm compared with vasocontraction. METHODS: Vasospasm was produced in the rabbit basilar artery by a two-hemorrhage method. Vasocontraction was induced by local application of KCl or serotonin to the rabbit basilar artery after a transclival exposure. The control animals were treated with saline instead of fresh blood. Serine/threonine protein phosphatase activity in the basilar artery was assayed with the use of [32P]phosphorylase-a as a substrate; protein phosphatase 1 activity was evaluated as protein phosphatase activity in the presence of 1 nmol/L okadaic acid, whereas protein phosphatase 2A activity was assessed as protein phosphatase activity inhibited by 1 nmol/L okadaic acid. RESULTS: Values of mean activity of protein phosphatase 1 in myofibrillar extract were 3.58 +/- 0.26 nmol/min per milligram in the control group, 3.22 +/- 0.12 nmol/min per milligram in the spastic group on day 2, and 3.01 +/- 0.16 nmol/min per milligram in the spastic group on day 4 (a significant decrease in protein phosphatase 1 activity in the spastic group on days 2 and 4). In contrast, these values did not show any significant changes in the KCl and serotonin groups. Values of mean activity of protein phosphatase 2A in cytosolic extract were 0.90 +/- 0.07 nmol/min per milligram in the control group, 0.75 +/- 0.10 nmol/min per milligram in the spastic group on day 2, and 0.62 +/- 0.17 nmol/min per milligram in the spastic group on day 4 (a significant reduction in protein phosphatase 2A in the spastic group on days 2 and 4). There was no evidence of significant changes of protein phosphatase 2A in cytosolic extract in the KCl and serotonin groups. CONCLUSIONS: Protein phosphatase 1 in myofibrillar extract is reported to catalyze the dephosphorylation of myosin light chain and calponin, whereas protein phosphatase 2A in cytosolic extract catalyzes the dephosphorylation of calponin and caldesmon. In addition, the phosphorylation of calponin and caldesmon results in the loss of their ability to inhibit smooth muscle contraction. Therefore, the significant decrease in activity of protein phosphatases 1 and 2A in vasospasm may result in uninterrupted vascular smooth muscle contraction by the preservation of phosphorylation of not only myosin light chain but also calponin and caldesmon.

Effects of protein kinase C modulators on multidrug resistance in human glioma cells.

To identify the role of protein kinase C (PKC) in multidrug resistance, the effects of phorbol-12-myristate-13-acetate (PMA), a PKC activator, or calphostin C, a PKC inhibitor, on intracellular vincristine accumulation and expression of P-glycoprotein phosphorylation were studied in one multidrug-resistant and three multidrug-sensitive human glioma cell lines. Basal PKC activities and immunoreactivities of PKC-alpha and -zeta were higher in multidrug-resistant cells than in multidrug-sensitive cells. There was no significant difference in the immunoreactivity of PKC-delta between multidrug-resistant and -sensitive cells, and immunoreactive PKC-beta, -gamma, and -epsilon were not detected in either multidrug-resistant or -sensitive cells. The treatment of multidrug-resistant cells with 100 nM PMA for 2 hours resulted in the activation not of PKC-zeta but of PKC-alpha, with concomitant decrease in vincristine accumulation and increase in P-glycoprotein phosphorylation. The exposure of multidrug-resistant cells to 100 nM PMA for 24 hours induced down-regulation not of PKC-zeta but of PKC-alpha, with concurrent decrease in vincristine accumulation, and reduced but still increased P-glycoprotein phosphorylation. The treatment of multidrug-resistant cells with 100 nM calphostin C for 2 hours decreased immunoreactive PKC-zeta and not immunoreactive PKC-alpha, inducing increase in vincristine accumulation, with concomitant decrease in P-glycoprotein phosphorylation. There was no evidence of significant change in vincristine accumulation in multidrug-sensitive cells treated with PMA or calphostin C. This may suggest that at least two isozymes of PKC, PKC-alpha and -zeta, are involved in P-glycoprotein phosphorylation and that vincristine efflux function in multidrug-resistant human glioma cells is closely associated with P-glycoprotein phosphorylation and is decreased by PKC inhibitor.

The immunohistochemical distribution of protein kinase C isozymes is altered in the canine brain and basilar artery after subarachnoid hemorrhage.

The effects of subarachnoid hemorrhage on immunoreactivities of protein kinase C isozymes (alpha, beta, gamma) were studied in canine brain and basilar artery. Mild and severe constriction of the basilar artery was shown 2 days and 7 days after hemorrhage, respectively. In control brain tissues, the isozymes showed distinct distributions and following hemorrhage, reactive astrocytes with protein kinase C alpha staining emerged in the surface of the pons and hippocampus. The control basilar artery showed protein kinase C alpha staining and its staining was decreased on day 7, consistent with the result of immunoblot. Our results demonstrated that subarachnoid hemorrhage induces gliosis with heavy protein kinase C alpha staining and down-regulation of protein kinase C in the artery.