Thin-filament-binding domains of cardiac and fast skeletal muscle troponin I isoforms as studied by epitope tagging

We examined the binding domains of cardiac and fast skeletal muscle troponin I (CTnI and FTnI, respectively) to myofibrils (MFs). Deletion mutants containing CTnI amino acid residues 1–79, 43–207 and 80–207 (CTnI-head, CTnI-tail-1 and CTnI-tail-2, respectively) and FTnI amino acid residues 1–54 and 55–182 (FTnI-head and FTnI-tail, respectively) were transiently expressed in cardiac and fast skeletal muscle cells. To monitor the intracellular localization of these exogenously introduced truncated TnIs, epitope tagging was used. CTnI-tail-1 was incorporated into cardiac MFs specifically, but CTnI-tail-2 was not assembled onto any MFs examined. This suggests that there is no potent actin filament-binding site in CTnI-tail-2. Since CTnI-tail-1 has an amino acid extension (CTnI residues 43–79) whose sequence is longer than that of CTnI-head-2; it appears that this sequence extension is important in binding to cardiac MFs. FTnI-tail, containing the inhibitory domain of actomyosin ATPase, showed intensive and specific incorporation into fast MFs. FTnI-tail was a homologous fragment of CTnI-tail-2, but the binding patterns of these two domains differed greatly from each other. It is possible that the absence of potent binding affinity of CTnI-tail-2 corresponding to the inhibitory domain of actomyosin ATPase is advantageous for continuous cardiac muscle contraction, since a potent inhibitory activity is a serious obstacle to cardiac muscle contraction. It can be assumed that distinctive binding ability of functional domains of TnI-tails reflect unique adaptations to muscles with different physiological properties.     

Efficacy of recombinant human granulocyte-macrophage colony-stimulating factor for chemotherapy-induced leukopenia in patients with non-small-cell lung cancer

To assess the feasibility and efficacy of rhGM-CSF in ameliorating chemotherapy-induced leukopenia in patients with advanced non-small-cell lung cancer, we conducted a double-blind placebo controlled phase III study in a multicenter setting. Patients were eligible if they had cytologically or histologically proven cancer, no prior chemotherapy, stage IIIB or IV disease, an Eastern Cooperativve Oncology Group (ECOG) performance status of 0–2, an age of less than 76 years, and no symptomatic brain metastasis, disseminated bone metastasis, or previous vertebral/pelvic irradiation. The chemotherapy regimen consisted of mitomycin given at 8 mg/m² on day 1, cisplatin given at 100 mg/m² on day 1, and vindesine given at 3 mg/m² i.v. on days 1 and 8 (MVP). If the granulocyte nadir count recorded after the first cycle of MVP was less than 1,000/mm³, patients were randomly assigned to receive recombinant human granulocyte-macrophage colonystimulating factor (rhGM-CSF) or placebo during the second cycle of MVP. The dose of rhGM-CSF was 125 g/m² given daily s.c. for 14 consecutive days starting on day 2. Of the 52 patients enrolled, 45 were evaluable. The nadir of granulocytes was significantly lower in the placebo group (P=0.007). The period during which the granulocyte count was less than 1,000/mm³ was significantly longer in the placebo group (median, 6 vs 10 days;P=0.04). The incidence of adverse effects related to rhGM-CSF, such as fever (38°C) and skin rash, was significantly higher in the rhGM-CSF group (P=0.011). The rate of response to chemotherapy did not significantly differ between the two groups. In conclusion, rhGM-CSF reduced the duration of chemotherapy-induced granulocytopenia. The clinical usefulness of this agent may be deminished because of the adverse effects encountered when it is used in combination with a moderately myelotoxic chemotherapy regimen.Institutions of the BI 71.018 Cooperative Study Group, National Cancer Center Research Institute (N. Saijo, Chairman): Douai Memorial Hospital (Y. Sano), Kanagawa Prefectural Cancer Center (K. Noda), Kantou Teishin Hospital (T. Uzawa), National Medical Center (J. Kabe), Shizuoka Prefectural General Hospital (H. Etou), Teikyo University (K. Mano), Tochigi Prefectural Cancer Center (Y. Saito), Tokyo Geriatric Medical Center (A. Kida), Tokyo Teishin Hospital (T. Morinari), Tokyo Metropolitan Komagome Hospital (S. Kudo), Tokyo Metropolitan Hiroo Hospital (A. Fujikawa), Toranomon Hospital (K. Nakada), Yokohama Municipal Hospital (K. Watanabe), National Cancer Center Central Hospital (K. Eguchi)     

Anti‐high mobility group box 1 monoclonal antibody ameliorates experimental autoimmune encephalomyelitis

High mobility group box 1 (HMGB1) is an established inflammatory mediator when released from cells. Recent studies have implicated extracellular HMGB1 in the pathogenesis of various autoimmune diseases. The objective of this study was to determine whether HMGB1 could be a therapeutic target for experimental autoimmune encephalomyelitis (EAE). In this study, an anti‐HMGB1 monoclonal antibody was injected intraperitoneally into a mouse model of EAE. We also measured serum cytokines levels in EAE and anti‐HMGB1 monoclonal antibody‐treated EAE. As a result, intraperitoneal injection of an anti‐HMGB1 monoclonal antibody ameliorated the clinical and pathological severity of EAE and attenuated interleukin‐17 up‐regulation in serum. In conclusion, HMGB1 is involved in EAE pathogenesis and could trigger inflammation in the central nervous system. The novel aspect of this study is the demonstration that anti‐HMGB1 ameliorates EAE. HMGB1 may be a novel therapeutic strategy for multiple sclerosis.     

Epithelioid myoepithelioma of the hard palate

Background

Myoepithelioma, a generally benign tumor comprised of myoepithelial cells, is an uncommon salivary gland tumor. Among four morphologic variants of myoepithelioma, epithelioid type has not been reported in the oral and maxillofacial region.

Case report

A 61-year-old man first noticed the mass 3 years previously. The oral examination revealed a firm, non-tender, and well-circumscribed mass in the middle of the hard palate. A magnetic resonance imaging scan showed a well-circumscribed mass with low signal intensity (T₁-weighted image) or increased signal intensity (T₂-weighted image).

Discussion

Immunohistochemically, the tumor cells in the present case reacted to the epithelial (CK HMW and CAM5.2) and the mesenchymal (vimentin) markers. However, myoepithelial markers (S-100 protein, α-smooth muscle actin, glial fibrillary acidic protein, and calponin), except p63, were not expressed in the tumor cells. These results indicated that the epithelial myoepithelioma cells differentiated into epithelial cells rather than myoepithelial cells. We believe that epithelioid myoepithelioma of the palate is a distinctive subtype of myoepithelioma that should be included in the differential diagnosis of tumors of the palate.