Autoepitopes on autoantigen centromere protein-A (CENP-A) are restricted to the N-terminal region, which has no homology with histone H3

Anti-centromere autoantibodies (ACA) are commonly found in the serum of patients with a limited type of scleroderma and other systemic autoimmune diseases. CENP-A is one of the major antigens against ACA and a histone H3-like protein. To analyse the autoantigenic epitopes of CENP-A, a series of truncated peptides of human CENP-A were expressed in Escherichia coli and immunoblotting analysis was performed with 91 ACA+ sera. Eighty sera (88%) with the ACA reacted to the 52-amino acids N-terminal region which is not homologous to H3, while no sera reacted to the C-terminus which has a sequence similarity with H3. Moreover, ELISA was also employed in this study using two synthetic peptides corresponding to the amino acid sequences 3-17 (peptide A) and 25-38 (peptide B). Peptides A and B were reactive to 78 (86%) and 79 (87%) of ACA, respectively. Core antigens of hepatitis B virus (HBV) and hepatitis C virus (HCV) have similar sequences to peptide A and/or peptide B, but three sera containing HBV without ACA and five sera containing HCV without ACA were found to be reactive to neither peptide. Centromere localization of CENP-A is dependent on the H3-like C-terminal domain which is not autoantigenic, while the antigenic N-terminal domain, which might play unidentified functional roles, should be an important region for the induction of ACA.     

Differences in specificities of anti-centromere sera for the monomeric and dimeric C-terminal peptides of human centoromere protein C

Centromere protein-C (CENP-C), one of the centromere autoantigens and components of the inner plate of the kinetochore, is suggested to make a dimer at the C-terminus. In order to investigate the presence of conformation-specific anti-centromere antibodies (ACA) to the dimer form, the C-terminal 124 amino acids (CF-124) were expressed in Escherichia coli, affinity purified and chemically cross-linked. Immunoblotting was utilized to compare the reactivities between the dimers and the monomers against 58 ACA(+) sera. The reactivities of the dimers were obviously higher in both IgG and IgM responses. The dimer was still more reactive than the glutathione S-transferase-fused monomer in some sera. Two kinds of CF-124 mutant (each contained one amino acid change at the N-terminal region of CF-124) and two cut segments of CF-124 (67 N-terminal amino acids and 58 C-terminal amino acids) were also examined. The former two mutants decreased the dimerization activity. The latter two mutants lost both activities except for the faint dimerization activity of the N-terminal half. Affinity-purified antibodies with CF-124 in a liquid phase containing the co-purified GroE protein of E. coli, GroEL, reacted to the centromere in culture cells. In conclusion, there are heterogeneous autoepitopes including some conformational epitopes at the C-terminal CENP-C.     

Alterations in the supraspinatus muscle belly with rotator cuff tearing: Evaluation with magnetic resonance imaging

We studied the supraspinatus muscle in patients with full-thickness rotator cuff tearing by analyzing its architecture on magnetic resonance imaging. Forty-six shoulders with rotator cuff tears requiring surgical repair were scanned parallel to the long axis of the supraspinatus muscle. We calculated the ratio of the largest width of the supraspinatus muscle belly to the distance from the greater tuberosity to the proximal end of the supraspinatus muscle. We classified the linear bands in the supraspinatus muscle, variation in its signal intensity, and irregularity and vagueness of its margins. The larger the rotator cuff tear, the more atrophic the supraspinatus muscle became. With increased atrophy, linear bands in the suprosupinotus muscle belly increased, muscle signal intensity became less uniform, and its margins became more irregular and vague.     

Significance of flow cytometric DNA analysis from freshly aspirated samples

The procedure of aspiration biopsy cytology by fine needle aspiration (ABC) is as option in establishing definitive diagnoses for breast cancers. In this series, a needle size of 21G was considered most suitable for ABC as well as flow cytometric DNA analysis. Histograms from fresh samples aspirated by fine needle clearly delineated a sharp peak in G&sup0;G(1) phases and also a better CV was obtained than with paraffin-embedded preparations. In addition, fresh samples gave more reliable DI and suggested the value of measuring nuclear DNA contents. It is believed that the prognoses of breast cancers are closely associated with DNA ploidy patterns. In this sense, flow cytometric DNA analysis of fresh samples of ABC is regarded as important in clinical use.     

Osteoplastic laminoplasty for cervical myeloradiculopathy secondary to ossification of the posterior longitudinal ligament

Summary Forty-seven patients with cervical myeloradiculopathy due to ossification of the posterior longitudinal ligament were treated by laminoplasty. The spinal canal from C3 to C7 was opened en bloc unilaterally and a spacer bone graft inserted to separate the floating laminae. The average follow up was 7.3 years (range 5 to 11 years). Favourable results were obtained in 35 patients and even though they had serious postoperative symptoms those with advanced neurological symptoms before operation showed considerable improvement. Late results were poor in patients who had greater than 50% compromise of the spinal canal by the ossified lesion. Laminoplasty is a safer method of obtaining favourable late results in patients with involvement of more than 3 vertebrae, but in those with a more severe compromise of the spinal canal by ossification, additional anterior decompression may be necessary later.

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Résumé Cet article présente les résultats à long terme de laminoplasties pratiquées pour myéloradiculopathie cervicale due à l'ossification du ligament vertébral commun postérieur, ainsi que les modifications observées dans les canaux rachidiens élargis. Habituellement le canal rachidien a été ouvert en bloc de C3 à C7, d'un seul côté, puis complété par une greffe osseuse afin de maintenir l'ouverture au niveau de la lame. On a utilisé la cotation de l'Association orthopédique japonaise, concernant la myélopathie cervicale, pour évaluer l'état des patients avant l'intervention chirurgicale, puis 2 ou 3 mois après la sortie et durant la période de surveillance. Les 47 patients de cette étude ont été suivis de 5 à 11 ans, soit en moyenne 7,3 ans. Les résultats finaux ont été favorables chez 35 patients et, même quand la symptomatologie postopératoire avait été assez préoccupante, les malades qui présentaient des symptômes neurologiques majeurs étaient notablement améliorés. Les résultats étaient mauvais chez les patients dont le rétrécissement du canal rachidien était supérieur à 50% (p<0.05). Il semble que la laminoplastie soit le moyen le plus sûr d'obtenir un résultat favorable mais une décompression antérieure complémentaire peut être nécessaire chez les patients atteints d'une sténose sévère due à une ossification du ligament vertébral postérieur.

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Presented in part at SICOT 93, Seoul, Korea, 28 August–3 September, 1993     

Lewy body-type degeneration in cardiac plexus in Parkinson's and incidental Lewy body diseases

Heart tissues of patients with PD or incidental Lewy body (LB) disease (ILBD) were examined by light and electron microscopy. LBs and alpha-synuclein-positive neurites were identified in the hearts from 9 of 11 patients with PD and from 7 of 7 patients with ILBD. LBs were present in both tyrosine hydroxylase-positive and -negative nerve processes, which are nerves of extrinsic sympathetic and intrinsic origin, respectively. These findings provide histologic evidence that the postganglionic sympathetic and intrinsic neurons in the heart are involved in the PD disease process.     

Roles of CD4+ and CD8+ T cells in discordant skin xenograft rejection

An essential role of murine CD4+ T cells in immune reactivity and skin graft rejection in discordant xenogeneic combinations have been reported. Our study was conducted to further clarify the roles of CD4+ and CD8+ T cells in discordant skin xenograft rejection, by using CD4 and CD8 knockout [C57BL/6 Cr Slc (B6; H-2b) background] mice. When human skins were grafted on CD8 knockout mice or B6 mice, both hosts rejected human skin grafts within 12 days after grafting. By contrast, survival of human skin grafts was significantly prolonged in CD4 knockout mice (mean survival times=19.3+/-(SD) 1.6 days; median 19 days). Fully allogeneic C3H/He Slc (H-2k) skin grafts were rejected within 14 days in CD4 knockout mice, suggesting that non-CD4+ T cells in CD4 knockout mice were immunocompetent for allograft rejection. In spleens of these recipient mice, CD8+ T cells seemed to be activated 10 days after human skin grafting. Immunohistological analysis revealed the infiltration of CD8+ T cells at the site of transplanted human skin on CD4 knockout mice. To further examine the role of CD8+ T cells in CD4 knockout mice, human skin grafting was performed on day 0 followed by administration of anti-CD8 monoclonal antibody on days 0, 5, and 14. The administration of anti-CD8 monoclonal antibodies caused the significant prolongation of human skin graft survival. These results indicate the following two conclusions: (1) CD4+ T cells have an essential role in rejecting discordant human skin xenografts rapidly and (2) however, CD8+ T cells also are capable of rejecting discordant human skin xenografts.     

Dietary Restriction Reduces the Prevalence of Osteonecrosis of the Caput Femoris in Spontaneously Hypertensive Rats

We investigated the effects of dietary restriction (DR), an experimental intervention known to suppress several strain-specific diseases, on the prevalence of osteonecrosis of the caput femoris in spontaneously hypertensive rats (SHR). At 6 weeks of age, the food intake of DR rats was restricted to 65% of the mean intake of control rats fed ad libitum (AL). Acute osteonecrosis of the caput femoris without reparative tissue response (RTR) was observed at 10 and 15 weeks in both DR and AL groups; no such acute lesion was seen at 20 and 30 weeks. The prevalence of osteonecrosis, osteonecrosis with/without reparative tissue response was significantly reduced in DR rats at 15 and 20 weeks, but not at 10 weeks. DR reduced the body weight by 30% and the length of the femur by 10%. Ossification of the caput femoris, known to be delayed in AL rats compared with Wistar Kyoto rats, was also restored by DR. Our results showed that dietary restriction reduced the prevalence of osteonecrosis and modulated the mechanical factors involved in the lesion. They also indicate that utilization of dietary restriction is a useful research tool for investigating the underlying mechanisms of osteonecrosis of the caput femoris in SHR. Received: 4 March 1997 / Accepted: 9 July 1998