Prospective study of kinesthesia after ACL reconstruction.

Previous studies used a variety of methods to assess kinesthesia, thus no consensus exists regarding kinesthetic adaptation after anterior cruciate ligament (ACL) reconstruction. This study prospectively examined whether kinesthesia is adapted after ACL reconstruction, and then discussed the actual angular velocity required to properly assess kinesthesia in ACL-reconstructed patients. 31 patients were evaluated using the threshold to detect passive motion (TTDPM) test, which was applied preoperatively, and at 3, 6, and 12 months following surgery. TTDPMs were measured at 15° or 45° of knee flexion toward both extension and flexion with angular velocities of 0.1°/s or 0.2°/s. ACL-reconstructed knees showed significantly impaired TTDPMs compared to healthy knees before the operation at 15° of knee flexion toward extension and at 45° of knee flexion toward both extension and flexion at 0.2°/s (15° of knee flexion toward extension, P=0.036; 45° of knee flexion toward extension, P=0.015; 45° of knee flexion toward flexion, P=0.030). However, there were no significant differences after 3 months of follow-up. On the basis of these results, applying 0.2°/s seems appropriate to assess TTDPM for patients with an ACL reconstruction, and kinesthesia is adapted within 12 months after the operation. Sensory function and biomechanical stability are also adapted following ACL reconstruction.

[Histopathological study of the temporal bones in patients with primary carcinomas of the ear].

The most common symptoms of patients with carcinomas of the middle ear or mastoid are otorrhea, facial paralysis, and hearing loss, including a sensorineural element and vertigo. The latter two symptoms are indicators of inner ear damage. However, few reports have been made concerning the histopathological changes that occur in the inner ear in the presence of a tumor. The present study was performed to determine the pattern of tumor invasion in the inner ear and the histopathological changes that occur in the inner ear in cases of ear carcinomas. Temporal bone sections from five patients (age: #39-73 years; 3 males and 2 females) who died from a primary carcinoma of the ear were studied histologically. The following features were examined: 1) localization of the tumor in the temporal bone, 2) pattern of tumor invasion in the inner ear, 3) pathological changes in the inner ear, including the cochlea, vestibule and semicircular canals. Tumor cells were still present in the temporal bone sections of all the patients except one, even though the patients had received various treatments for the carcinoma, including radiation therapy, surgery and chemotherapy. Marked inflammatory and necrotic changes were observed in cases where the tumor had invaded the external auditory canal, middle ear cleft, internal auditory canal, and in some cases the inner ear. In cases where the tumor invaded the inner ear via the internal auditory canal rather than directly from the middle ear, the otic capsule is thought to have acted as a barrier against tumor invasion. In addition, marked degenerative changes throughout the entire inner ear structures were noted. These changes may have arisen from an attenuated blood supply to the inner ear as a result of pressure from the tumor in the internal auditory canal, tumor infiltration of the labyrinthine artery.

Cochlear neuronal loss are determined by use of a neurofilament protein antibody in cases of bilateral profound deafness.

The temporal bones of two patients with profound bilateral deafness from infancy were studied immunohistochemically, using a neurofilament protein antibody to detect the cochlear neuronal elements. One patient exhibited Mondini dysplasia of the inner ear, with the organ of Corti almost completely deteriorated. The other patient is the first reported case involving complete aplasia of the organ of Corti in all turns. In both cases, the immunohistochemical staining clearly revealed a severe reduction in the number of afferent neurons, such as dendrites, spiral ganglion cells and cochlear axons. The number of efferent spiral bundles in the osseous spiral lamina and intraganglionic portion also decreased in parallel with the reduction in the number of cochlear afferent neurons. Our results are inconsistent with previously reported cases of presbycusis and acquired deafness induced by the measles virus, in which efferent neurons were preserved while afferent neurons degenerated. The loss of both the efferent and afferent neurons might be characteristics of congenital deafness.

Matrix metalloproteinase digestion of aggrecan in human cartilage tumours.

Substantial experimental and clinical evidence suggests that the catabolism of extracellular matrix components is a prerequisite for invasive and metastatic behaviour of solid tumours. Chondrosarcomas are malignant cartilaginous tumours that most commonly arise in bone, and the large aggregating proteoglycan aggrecan is a major component of the extracellular matrix of these tumours. Matrix metalloproteinases (MMPs) have been implicated in tumour invasiveness. The purpose of this study was to determine whether MMPs play a role in aggrecan catabolism in cartilage tumours. In order to detect aggrecan digestion products resulting from in vivo cleavage at the MMP site, protein extracts from human articular cartilage and from various cartilage tumours were analysed by Western blot using an antibody to the FVDIPEN neoepitope generated by MMP cleavage. Examination of cartilage extracts revealed a trend of increasing aggrecan digestion at the MMP site with age. One hyaline chondrosarcoma and three osteochondromas lacked detectable aggrecan fragments with the carboxy terminal FVDIPEN neoepitope. Two osteochondromas gave weak signals. However, all chondrosarcomas with degenerating extracellular matrix or with a myxoid component exhibited strong FVDIPEN immunoreactivity. These results demonstrate that, in contrast to the benign cartilage tumour osteochondroma, human chondrosarcomas contain abundant aggrecan degradation products resulting from cleavage in vivo at the MMP site in the interglobular domain. These data support the concept that MMPs participate in the degradation of extracellular matrix in chondrosarcoma, allowing the neoplastic chondrocytes to escape local confinement, migrate, and invade neighbouring and remote tissues.

[Histopathological studies of temporal bones of patients with malignant melanoma].

Although malignant melanoma is known to metastasize to various sites including the temporal bones, there have been few studies on temporal bone histopathology in malignant melanoma. Here we describe the temporal bone histopathology of 5 patients (10 temporal bones) who died of malignant melanoma with multiple metastasis to many internal organs and bones. We investigated the temporal bone based on the following three points: 1) the presence of metastatic lesions in the temporal bone, 2) inner ear pathology, and 3) the distribution of melanin in the inner ear. Normal melanin distribution was also studied in 35 temporal bones of patients without malignant melanoma. Metastatic malignant melanoma was observed in 5 temporal bones from 3 patients, in two of whom the internal auditory canal was involved bilaterally by melanoma cell infiltration. In the remaining patient (one ear), metastatic melanoma was found along the dura mater of the posterior cranial fossa to the mastoid air cells. In the former two patients, the inner and outer hair cells as well as the stria vascularis showed degenerative changes to various extents. In particular, the inner ear changes were severe in the ear with the decongestion of the inner ear vessels. Melanin was found in the modiolus, stria vascularis, osseous spiral lamina, membranous labyrinth, and endolymphatic sac, as previously reported. The amount of melanin in the inner ear increased with age in the control patients, but was greater than in the controls, in all of the cases of malignant melanoma except one, in which metastatic lesions were present in the internal auditory canal with marked congestion of the inner ear vessels.

Analysis of giant cell tumor of bone with pulmonary metastases.

The authors reviewed 6 cases of giant cell tumor of bone with pulmonary metastases, analyzed the growth rate of the metastases, and performed flow cytometry on paraffin blocks from primary and metastatic lesions. Surgery on the pulmonary metastases was done in 3 cases. Chemotherapy was administered in all 6 cases. If the doubling time of the pulmonary metastases was more than 80 days, the case was considered to be slow growing or in regression. However, if the doubling time was approximately 30 days, the prognosis was poor. The cases were divided into 3 types according to the prognosis. After chemotherapy (cyclophosphamide), the pulmonary metastases decreased in size in 2 cases. In 2 other cases, the metastases were slow growing, and the remaining 2 patients died early. Flow cytometric analysis was performed on primary and metastatic lesions, which showed a diploid pattern in most cases, but a tetraploid pattern in 1 recurrent case. Pulmonary metastatic lesions displayed a diploid pattern in 4 cases from which materials were taken either at surgery or autopsy.

The role of transforming growth factor-beta in PEG-rHuMGDF-induced reversible myelofibrosis in rats.

Pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) injected at a suprapharmacologic dose (100 microg/kg) daily for 5 d in normal rats caused marked increases in marrow megakaryocytes and platelet counts at 6-8 d followed by gradual decreases to control levels at 10-20 d. Interestingly, in addition to the expected thrombopoiesis, PEG-rHuMGDF was associated with myelofibrosis with a predominance of reticulin fibres at day 10 followed by complete normalization by day 20. At 6-8 d, the levels of transforming growth factor-beta1 (TGF-beta1) in the extracellular fluid of the marrow, the platelet poor plasma, and the platelet extract were increased 23-, 7- and 2-fold, respectively. The elevated levels of TGF-beta1 were gradually reduced to baseline levels at 13-20 d in accordance with the normalization of myelofibrosis and thrombopoiesis. An ultrastructural analysis showed that large fragments of megakaryocytes were deposited in the marrow parenchyma of PEG-rHuMGDF-treated rats at day 6. PEG-rHuMGDF administration at pharmacologic doses (1 and 10 microg/kg) did not induce the deposition of reticulin fibres in the marrow. These findings suggest that TGF-beta1 leaked from megakaryocytes is involved in the development of the PEG-rHuMGDF-induced myelofibrosis and that this is a reversible process related to the regulation of the excess production of platelets.

Radical radiotherapy for T3 laryngeal cancers.

PURPOSE: Evaluation of the local control rate and voice preservation probability in radically irradiated patients with T3 laryngeal cancer. PATIENTS AND METHODS: From 1974 through 1992, 37 previously untreated patients with T3 laryngeal cancer (supraglottic 15, glottic 22) were treated with initial radical radiotherapy and surgery for salvage. RESULTS: Two-year local control rate with radiotherapy alone, ultimate voice preservation rate, and ultimate local control rate for T3 supraglottic cancer were 33%, 33%, and 60%, respectively. Corresponding figures for T3 glottic cancer were 32%, 23%, and 77%, respectively. Five-year cause-specific survival rate for T3 supraglottic cancer and glottic cancer were 47% and 77%, respectively. In T3 supraglottic cancer, none of the 4 patients with subglottic tumor extension attained local control by radiotherapy alone, and local-regional recurrence-free time were significantly shorter in patients with subglottic tumor extension or tracheostomy before radiotherapy. There were no serious late complications such as chondronecrosis, rupture of carotid artery attributed to radical radiotherapy, while 3 patients had severe laryngeal edema requiring total laryngectomy. CONCLUSION: Current results indicated that initial radical radiotherapy and surgery for salvage might be an option for some patients with T3 laryngeal cancer.

[Clinical characteristics of chronic sinusitis with different lower respiratory tract diseases].

We determined the clinical characteristics of chronic sinusitis in patients with different lower respiratory tract diseases. Sinusitis was divided into three groups. The first group was sinusitis with bronchial asthma. The second group was sinusitis with chronic bronchitis, bronchiectasis or diffuse panbronchiolitis. This type of sinusitis is generally called sinobronchial syndrome (SBS). The third group was sinusitis without lower respiratory tract disease. Because the pathogenesis of these lower respiratory tract diseases differs with the diseases, the pathogenesis of sinusitis accompanied by these lower respiratory tract diseases also may be different. Clinical and laboratory examinations used in this study were rhinoscopy, X-ray examination of the paranasal sinuses, cytological study of nasal secretion and the saccharin test for the muco-ciliary function of the nasal mucosa. By rhinoscopic examination, no difference was found in the size of the nasal polyp among three groups. The results of other examinations were as follows. X-ray examination: Involvement of the ethmoid sinuses was greater than that of the maxillary sinuses in the patients with sinusitis with bronchial asthma. In contrast, involvement of the maxillary sinuses in SBS patients was greater than that of the ethmoid sinuses. Cytology of nasal secretion: Dominant inflammatory cells in the patients with sinusitis with bronchial asthma were eosinophils, while neutrophil were more frequently found in the SBS patients. Saccharin test: Most of the patients with bronchial asthma showed normal responses. In the SBS patients, however, only a few patients showed a normal response and the others showed prolonged responses. Clinical characteristics of the patients with sinusitis without lower respiratory diseases were more similar to those of the SBS patients. In conclusion, there were distinct differences in these clinical characteristics between sinusitis with bronchial asthma and SBS. These results suggest that the pathogenesis or the inflammatory process of sinusitis is heterogeneous and whether the inflammation is allergic or not may be important.

Such hydrophobic peptides as dansylated mastoparan can elevate the fertilization membrane of sea urchin eggs.

Melittin is known to be a major hydrophobic peptide component in honeybee venom that can cause as much elevation of fertilization membrane of sea urchin eggs as normal fertilization. The action of melittin has been thought to be closely related with its ability to facilitate the phospholipase A2 activity on the eggs. However, another peptide "mastoparan" from wasp venom was not found here to cause any elevation of the membrane, although it can activate the enzyme as well as melittin. On the other hand, mastoparan was found to get the membrane-elevating activity only when its amino groups were modified with hydrophobic substituents. N epsilon-Substituted mastoparan with a dansyl group in Lys11 residue was most effective among the analogs examined here. Our findings indicate that the facilitation of phospholipase by the peptides have little relation with the membrane generation. Such hydrophobic moiety as the dansyl group in the peptides must cause the cortical reaction on the eggs in cooperation with peptide moiety. The dansylated peptide will be a useful tool to induce the artificial fertilization of sea urchin eggs.

[Histopathological studies on paranasal mucosa from patients treated with erythromycin].

Low-dose and long-term administration of erythromycin (EM therapy) has been reported to be very effective for patients with intractable chronic sinusitis including sinobronchial syndrome. However, we sometimes encounter patients whose sinusitis is extremely resistant to EM therapy. Therefore, the present study was carried out to determine the correlation between the clinical efficacy of EM therapy and the histopathological features of the ethmoidal mucosa and nasal polyps of patients treated with erythromycin. Patients with significant lymphocytic infiltration in the submucosal area responded well to EM therapy. Furthermore, patients with neutrophilic infiltration within and beneath the ciliary epithelium tended to show improvement with this therapy. On the other hand, EM therapy was minimally effective in patients whose subepithelial layer showed marked eosinophilic infiltration. In patients showing excellent and good clinical results, histological changes in the paranasal mucosa after therapy were as follows: a decrease in the number of inflammatory cells, reduced interstitial edema, increased fibrosis and normalization of the secretory glands. From these results, we conclude that erythromycin may suppress chronic inflammation except for an allergic reaction characterized by marked eosinophilic infiltration.

[Growth rate analysis of pulmonary metastases in giant cell tumor of bone].

We reviewed six cases with pulmonary metastases in giant cell tumor of bone. The growth rate and the doubling time of pulmonary metastases was measured and compared to the clinical course. The six cases were classified into two each of the following three types: 1) spontaneous remission, 2) continuously slow growing and 3) rapid growing. A favorable factor was a doubling time of more than 80 days in pulmonary metastases. A poor prognosis was indicated by a doubling time of around 30 days. Aggressive primary lesions should be excised widely.

Erythromycin therapy for otitis media with effusion in sinobronchial syndrome.

Chronic sinusitis is frequently associated with chronic lower respiratory tract diseases, and the association is referred to as sinobronchial syndrome (SBS). This study was carried out to determine the incidence of otitis media with effusion in the patients with sinobronchial syndrome and to investigate the efficacy of low-dose and long-term erythromycin therapy for otitis media with effusion associated with sinobronchial syndrome. We have found a high incidence of otitis media with effusion in patients with sinobronchial syndrome, the morbidity rate being 54% in 50 cases studied. This ear disease seems to be the major cause of hearing disturbance in patients with sinobronchial syndrome. Sixteen patients with both sinobronchial syndrome and otitis media with effusion were given low-dose and long-term erythromycin therapy (erythromycin base, 600 mg/d for more than 4 months); of these, 13 became effusion-free and most subjects showed improvement in the symptoms of sinobronchial syndrome. The erythromycin therapy thus seems to be exceedingly effective for the treatment of sinobronchial syndrome and associated otitis media with effusion.

Chronic tonsillitis and IgA nephropathy. Clinical study of patients with and without tonsillectomy.

To determine whether tonsillectomy is a significantly effective treatment in the clinical course of IgA nephropathy, we did a comparative study on 50 patients with IgA nephropathy and chronic tonsillitis. We divided the patients into two groups: 35 patients with and 15 without tonsillectomy (control group). With or without tonsillectomy, renal function became progressively worse during the follow-up period in most patients with a serum creatinine level of > 1.4 mg/dl at the time of renal biopsy. In patients with a serum creatinine level of < or = 1.4 mg/dl, renal function remained normal in all subjects with tonsillectomy, but worsened in 3 patients out of 13 without tonsillectomy. Improvement in proteinuria/hematuria was found more frequently in the tonsillectomized group than in the controls. Furthermore, the serum IgA level was significantly reduced after tonsillectomy, especially in patients showing improvement. From these results we conclude that tonsillectomy was effective for patients with IgA nephropathy complicated by tonsillitis when the operation was performed before deterioration of renal function.

Erythromycin inhibition of lipopolysaccharide-stimulated tumor necrosis factor alpha production by human monocytes in vitro.

The mechanism of clinical effectiveness of low-dose and long-term erythromycin (EM) treatment for diffuse panbronchiolitis, sinobronchial syndrome, and associated otitis media with effusion was investigated by studying the effects of EM on tumor necrosis factor alpha (TNF-alpha) production by cultured human monocytes stimulated with lipopolysaccharide. At concentrations of 0.1 microgram/mL or more, EM inhibited TNF-alpha release from human monocytes stimulated by lipopolysaccharide in a dose-dependent manner. Of the other macrolides tested, roxithromycin, an EM derivative, also showed significant inhibition of TNF-alpha production, whereas josamycin failed to inhibit TNF-alpha release from monocytes. Nonmacrolidic drugs such as minocycline hydrochloride, ofloxacin, or penicillin G had no significant effect on TNF-alpha production. These results suggest that the clinical improvement of chronic respiratory diseases by EM may depend on the suppression of production of inflammatory cytokines such as TNF-alpha.

[Human monocytes show chemotaxis in response to cholesteatoma debris].

Using a microchamber technique, we tested cholesteatoma debris and certain of its constituents for effects on the migration of human peripheral blood monocytes and polymorphonuclear leukocytes. Cholesteatoma debris induced significant migration of monocytes. When the individual constituents of cholesteatoma debris, i.e., alpha-keratin, cholesterol, lauric acid and lipopolysaccharides, were tested for monocyte chemotaxis, only alpha-keratin induced significant monocyte migration. alpha-keratin extracted from the cholesteatoma debris with 8 M urea also induced migration of monocytes with a bell-shaped dose-response curve, which is frequently encountered with chemoattractants. Therefore, cholesteatoma debris and one of its components, alpha-keratin, are potent chemoattractants for human monocytes. On the other hand, cholesteatoma debris showed no significant chemotactic effect on polymorphonuclear leukocytes. Based on the present and our previous results, cholesteatoma debris acts on monocytes/macrophages as a strong chemotactant, a potent activating (priming) factor, and an inducer of production of tumor necrosis factor, which is a bone-resorbing cytokine. Therefore, we concluded that macrophages induced by cholesteatoma debris may play an important role in the pathogenesis of bone resorption in cholesteatoma otitis.

Cholesteatoma debris as an activator of human monocytes. Potentiation of the production of tumor necrosis factor.

Tumor necrosis factor (TNF) is a cytokine which stimulates osteoclastic bone resorption and inhibits collagen synthesis in vitro. In this study the effect of human cholesteatoma debris and its constituents on the production of TNF-alpha by human monocytes in vitro was studied. Cultured human peripheral monocytes secreted TNF into the culture medium when exposed to cholesteatoma debris in a dose-dependent manner. The TNF production, however, was partially inhibited by the treatment of the debris with polymyxin B which inhibits biological activities of lipopolysaccharide (LPS). When individual constituents of cholesteatoma debris, i.e. keratin, cholesterol, lauric acid and LPS, were added to the cultured monocytes at concentrations equivalent to those in the debris, significant production of TNF was observed only with the keratin and LPS. These data suggest that cholesteatoma debris is a potent activator of the TNF production of human monocytes in vitro, and that LPS and keratin are responsible for the production.

Activation of peritoneal macrophages with human cholesteatoma debris and alpha-keratin.

The effect of human cholesteatoma debris on mouse peritoneal macrophages was studied in vivo. The number of macrophages and lymphocytes increased 5 days after injection of the debris into the peritoneal cavity. A similar increase in peritoneal cells was observed when an urea-extracted fraction of the cholesteatoma debris or alpha-keratin, a major component of the debris, was injected. Both cholesteatoma debris- and alpha-keratin-elicited macrophages exhibited a greater response of luminol-dependent chemiluminescence upon exposure to zymosan, suggesting that the elicited macrophages were activated. In contrast, other constituents of the debris, such as cholesterol or fatty acid--with the exception of lipopolysaccharide (LPS)--failed to elicit or activate peritoneal macrophages at the similar doses detected in the debris. The chemiluminescent response of macrophages obtained by injecting LPS was, however, much lower than that of alpha-keratin-induced macrophages. These results indicate that cholesteatoma debris is capable of eliciting and activating macrophages, and that alpha-keratin is responsible for the activation.

The mitotic apparatus-associated 51-kDa protein from sea urchin eggs is a GTP-binding protein and is immunologically related to yeast polypeptide elongation factor 1 alpha.

We investigated the biochemical characteristics of the 51-kDa protein that is a major mitotic apparatus-associated basic protein of sea urchin eggs (Toriyama, M., Ohta, K., Endo, S., and Sakai, H. (1988) Cell Motil. Cytoskeleton 9, 117-128). The amino acid composition of the 51-kDa protein was apparently different from those of tubulin, actin, histones, and myelin basic protein; yet it was similar to those of polypeptide elongation factors 1 alpha (EF-1 alpha). In addition, antibody to EF-1 alpha from yeast cross-reacted with the 51-kDa protein. [3H] GTP binding activity was detected in the phosphocellulose-purified fraction (PC fraction) which predominantly contained the 51-kDa protein and was shown to be specific to GTP, GDP, guanylyl imidodiphosphate, and ITP. Photo-affinity labeling using [alpha-32P]8-azidoguanosine triphosphate (8-azido-GTP) demonstrated that a 51-kDa polypeptide in the PC fraction specifically bound 8-azido-GTP. This GTP-binding polypeptide was bound to a GTP affinity column, could be eluted by the addition of GTP, and was immunoreactive with anti-51-kDa protein antibodies. When the PC fraction was applied to a gel filtration chromatography column, GTP binding activity was completely coeluted with the 51-kDa protein. Furthermore, the PC fraction and the gel filtration-purified fraction had EF-1 alpha activity: [14C]Phe-tRNA transferring activity to ribosomes in the presence of poly(U) and ribosome-dependent GTPase activity. The results indicate that the mitotic apparatus-associated 51-kDa protein is a GTP-binding protein and suggest that it is structurally and functionally related to yeast EF-1 alpha.

Purification and characterization of a Ca2+-dependent actin filament severing protein from bovine adrenal medulla.

We describe the purification of an actin regulatory protein from bovine adrenal medulla. This protein caused a dose-dependent decrease of the specific viscosity of actin solution within 30 s of its addition in a Ca2+-sensitive way. Sedimentation assays and the observation by electron microscopy showed that this effect was ascribable to the fragmentation of actin filaments. This protein apparently promoted nucleation of actin polymerization and increased the critical concentration of actin for polymerization nearly 5-fold, suggesting its binding to the barbed end of actin filaments. The inhibitory effect of this protein on the elongation of actin from the barbed end of the myosin subfragment S1-labeled actin seeds confirmed this suggestion. These properties are similar to those of gelsolin. However, the physicochemical properties of this protein having a single polypeptide chain with a molecular weight of 74,000, a Stokes radius of 3.9 nm, a sedimentation coefficient (s0(20),w) of 4.5 S, and an immunological characterization showed that this protein is different from gelsolin.