Dynamic movement of the long head of the biceps tendon in frozen shoulders.

PURPOSE: To assess the importance of the dynamic movement of the long head of the biceps (LHB) tendon for treatment of frozen shoulder. METHODS: 87 consecutive patients (with 88 frozen shoulders) aged 36 to 77 (mean, 54) years underwent arthroscopic capsular release by a single surgeon. Preoperative treatments included rehabilitation, steroid and/or hyaluronic acid injections. The inclusion criteria were severe night pain, no improvement of flexion and external rotation, and poor response to rehabilitation for at least 6 months. Shoulders were divided into 3 types; types A/B/C indicate slight/moderate/severe degree of synovitis and adhesion of the LHB tendon to the rotator interval. 23 shoulders were type A, 26 type B, and 39 type C. 18 of the 39 type-C shoulders were controls with release of the capsule alone but not the LHB tendon. Patients were followed up for a mean of 21 (range, 12-35) months. Changes in the American Shoulder and Elbow Surgeons (ASES) score, range of movement, and muscle strength (flexion and external rotation) among types A, B, C, and controls were compared. RESULTS: The severity of the adhesion of the LHB tendon to the rotator interval was associated with the ASES score. In all adhesion types, muscle strength and the range of movement in flexion, external rotation, and internal rotation improved postoperatively. CONCLUSIONS: Arthroscopic capsular release for adhesion of the LHB tendon to the rotator interval improves the sliding movement and thereby shoulder function.

Effects of protein kinase inhibitors on the cell motility stimulated by autocrine motility factor.

Preincubation of Dunn osteosarcoma cells for 1 h with both 100 nM of staurosporine and 10 micrograms/ml of genistein resulted in a significant decrease in the motility stimulated by autocrine motility factor (AMF), whereas these reagents did not affect the basal motility and proliferation at these concentrations. The effect of the agents on the stimulated motility was both dose- and time-dependent. The motility stimulated by the anti-AMF receptor mAb was also inhibited. In contrast, H-8 had a negligible effect upon the stimulated motility. These data suggest that both kinase C and tyrosine kinase play a role in AMF-stimulated cell motility, while protein kinase A, which is selectively associated with the adenylate cyclase pathway, may not be required for the stimulation.

Differential purification of autocrine motility factor derived from a murine protein-free fibrosarcoma.

We have previously shown that a protein-independent growing fibrosarcoma, Gc-4 PF has a high motile response to its cultured medium, which is associated with an increase in expression of gp78, a cell surface receptor for autocrine motility factor (AMF). Here we show that the cultured medium contains two motile activities, acidic and basic AMFs with regard to binding features on ion exchange chromatography. These two AMFs were purified by sequential DEAE anion exchange, CM cation exchange, and gel filtration chromatographies. However, both acidic and basic AMFs have a similar size of 55 kDa and 65 kDa under non-reducing and reducing conditions, respectively, with the same pI of 6.5. The stimulated motility of both AMFs was inhibited by the pertussis toxin (PT), but not by Streptomyces hyaluronidase. These two AMFs significantly stimulated the lung colonizing properties of the self-producing cells by 1.5-fold. These results suggest that both acidic and basic AMFs may correspond to the previously reported AMF and confirm directly that the AMF-gp78 signaling pathway is involved in cell motility associated with metastatic property.

Different types of DNA cleavage involved in osteosarcoma cell death.

To induce cell death in osteosarcoma, a murine osteosarcoma clone, DOS C14 was exposed to: i) topoisomerase II-reactive epipodophyllotoxin, etoposide (ETO); ii) glucocorticoid analogue, dexamethasone (DEX); and iii) ultraviolet light (UV) irradiation. In MTT assay, fifty micromolar ETO, 100 mu M DEX and UV irradiation for 90 min reduced the cell number to 20% of that of the control. The cytotoxic effects of ETO and DEX were dose-dependent, while those of UV irradiation were time-dependent. Endonuclease cleavage of DNA into internucleosomal fragments was not recognized on DOS C14 osteosarcoma treated with 50 mu M ETO, 100 mu M DEX or UV irradiation. Aurintricarboxylic acid (ATA) also failed to inhibit the reduction in the number of viable cells. However, DNA from DOS C14 osteosarcoma cells exposed to 1 h UV irradiation showed smearing DNA fragments after treatment with S1 endonuclease, while such single strand modification was not detected in DNA extracted from cells exposed to ETO or DEX. On the other hand, pulse field gel electrophoresis revealed that cleavage of DNA into high molecular weight fragments estimated as 50-150 kilobase pairs (kbp) with a peak of 100 kbp was found in DOS C14 osteosarcoma cells exposed to 50 mu M ETO and 100 mu M DEX. The cytotoxicity of ETO and DEX was reduced by okadaic acid, while UV-induced cytotoxicity was not reduced by okadaic acid. Furthermore, okadaic acid inhibited the formation of high molecular weight DNA fragments in a dose-dependent manner. These results suggest that two types of DNA degradation exist in osteosarcoma death; one is random breakage of DNA, and the other is large DNA fragmentation, which may be produced by an activation of putative ATA-insensitive and okadaic acid-sensitive endonuclease.

Autonomic proliferation of 2 distinct protein-free tumor-cell lines depends on the calmodulin pathway.

Four calmodulin antagonists (W-5, W-7, W-12 and W-13) specifically inhibited the proliferation of two protein-free sublines (P815 PF and Ehrlich PF), although they did not inhibit that of their serum-dependent counterparts. This inhibition of proliferation paralleled the inhibiting activity of the four antagonists against calmodulin. Inhibiting activity of the agents against protein-free cells was cancelled by supplementation of the culture medium with serum. It is suggested that plural signalling pathways switched 'cross-overly' play an important role in the proliferation of protein-free cells. Immunoblotting showed that the soluble calmodulin content in cytoplasm is not changed in protein-free sublines as compared with serum-dependent lines. However, nuclear calmodulin levels of the two protein-free subclones were lower than those of their serum-dependent counterparts. It is suggested that intranuclear calmodulin rather than that in cytoplasm is more important in autonomic proliferation. The internal 'replicon hypothesis' may explain how this autonomic proliferation depends on the nuclear calmodulin pathway.

AB3217-A, a novel anti-mite substance produced by a strain of Streptomyces platensis.

AB3217-A, a novel anti-mite substance, was isolated from the fermentation broth of a streptomycete strain. The strain was isolated from a soil collected at Kita-azumi, Nagano Prefecture, Japan, and identified as Streptomyces platensis AB3217. AB3217-A was purified by Amberlite IR120B, Diaion HP-20 and CM-Sephadex C-25 column chromatographies. The molecular formula was determined as C17H23NO7 by elemental analysis, MS and 13C NMR spectroscopy. The structure of AB3217-A was determined to be (1R,3S,4S,7R,8R,11R,12S,13R)-4,12,13-trihydroxy-8-(4-methoxy phenyl)-6-aza-2,9,14-trioxatricyclo-[9.2.1.0(3,7)]tetradecane by spectroscopic analysis and X-ray crystallographic analysis. The molecule of AB3217-A has unique structure that deacetylanisomycin and beta-D-xylofuranose linked through glycosidic bond and ether bond resulting in the formation of nine-membered ring. AB3217-A showed marked activity against the two spotted spider mite, Tetranychus urticae.

Synovectomy reduces stromal-cell-derived factor-1 (SDF-1) which is involved in the destruction of cartilage in osteoarthritis and rheumatoid arthritis.

We have compared the concentrations of stromal-cell-derived factor-1 (SDF-1), matrix metalloproteinase-1 (MMP-1), MMP-9 and MMP-13 in serum before and after synovectomy or total knee replacement (TKR). We confirmed the presence of SDF-1 and its receptor CXCR4 in the synovium and articular cartilage by immunohistochemistry. We established chondrocytes by using mutant CXCR4 to block the release of MMPs. The level of SDF-1 was decreased 5.1- and 6.7-fold in the serum of patients with OA and RA respectively, after synovectomy compared with that before surgery. MMP-9 and MMP-13 were decreased in patients with OA and RA after synovectomy. We detected SDF-1 in the synovium and the bone marrow but not in cartilage. CXCR4 was detected in articular cartilage. SDF-1 increased the release of MMP-9 and MMP-13 from chondrocytes in a dose-dependent manner. The mutant CXCR4 blocked the release of MMP-9 and MMP-13 from chondrocytes by retrovirus vector. Synovectomy is effective in patients with OA or RA because SDF-1, which can regulate the release of MMP-9 and MMP-13 from articular chondrocytes for breakdown of cartilage, is removed by the operation.

Fracture of the posterior medial tubercle of the talus treated by internal fixation: a report of two cases.

A fracture through the posterior medial tubercle of the talus is quite rare. Although excision of the bone fragments has been reported previously in this fracture, cases of internal fixation of the posterior medial tubercle of the talus have not been reported. Two patients are described who presented with a fracture through the posterior medial tubercle of the talus which was treated with internal fixation. Our patients appeared normal on physical examination and returned to work, with no evidence of avascular necrosis of the fragments.

[Percutaneous transabdominal whole layer core needle biopsy for staging the urinary bladder cancer].

Accurate staging is one of the most important determinants necessary in planning an effective treatment for bladder cancer, but at the present time, staging by the conventional clinical methods are not completely reliable. Accordingly, we have newly developed a percutaneous transabdominal core needle biopsy technique capable of obtaining good cylindrical specimens of the entire layer. In order to obtain the core specimen of the bladder wall, which is a vesico-elastic material, the biopsy needle must penetrate the bladder wall at an extremely high speed. We have developed an original automatic biopsy instrument and modified the head of needle for this purpose. With this method, we could obtain good core specimens for pathological staging in about 90% cases. The correlation between the pathological stagings of core specimens and those of cystectomy specimens were in good agreement. This technique is accurate not only for preoperative pathological staging, but also monitoring histopathologically the responsiveness of multi-disciplinary treatment for invasive bladder cancer. This method allows us to determine the optimal therapeutic modality for managing individual patient with invasive bladder cancer.

[Prophylaxis of superficial bladder tumor with the intravesical instillation of bacillus Calmette-Guerin or anti-cancer agents. A retrospective study].

It was retrospectively analyzed whether intravesical instillation of bacillus Calmette-Guerin (BCG) or anti-cancer agents had prophylactic effect or not after removal of superficial bladder tumors. The results over a follow-up period ranging from 6 to 40 months showed 23.8 per cent recurrence in group 1 patients treated with BCG (21 patients), 46.7 per cent recurrence in group 2 treated with anti-cancer agents (45 patients) and 52.3 per cent recurrence in the group 3 (127 patients) which were not received any intravesical drugs. Long-term results among the 3 groups calculated with Kaplan-Meier method demonstrated that the instillation of BCG or of anti-cancer agents was more useful for prophylaxis, compared with the actuarial non-recurrence rates of group 3. The instillation of BCG showed good prophylactic effects especially in recurrent, grade 2 and pTa bladder tumors. The instillation of anti-cancer agents showed to provide prolonged protection from recurrence, but the instillation of BCG did not.

[Thermotherapy for cancer of the urinary bladder in combination with tegafur and picibanil--with special reference to the serum bladder and bladder cancer tissue concentrations of tegafur in a perfusion fluid].

Thermotherapy combined with Tegafur and Picibanil was performed in 32 patients with cancer of the urinary bladder. The thermotherapy was performed with a closed circulation type of thermotherapeutic apparatus manufactured for trial, using a 3-way catheter. The flow quantity of the perfusate was 150 ml/min at a constant temperature of 43 degrees C. The temperature was monitored at the inlet and outlet catheter at the urethral meatus. The thermotherapy was performed for 6 hours once a week. Tegafur and Picibanil were added to the perfusate at the time of thermotherapy. No anesthesia was used. Complete disappearance of tumor occurred in 9 cases. The concentration of Tegafur in the serum, tumor and bladder wall was simultaneously measured. The serum levels after two hours of thermotherapy with 16 gm of dissolved Tegafur showed that FT-207 was 28.837 mcg/ml and 5-FU, 0.028 mcg/ml. After 4 hours the FT-207 was 36.464 mcg/ml and 5-FU, 0.040 mcg/ml. By the 6th hour FT-207 was 39.430 mcg/ml and 5-FU, 0.049 mcg/ml. After 24 hours FT-207 was 11.242 mcg/ml and 5-FU, 0.013 mcg/ml in the serum, while FT-207 was 8.205 mcg/g and 5-FU, 0.274 mcg/g in the bladder tumor, and FT-207 was 17.029 mcg/g and 5-FU, 0.157 mcg/g in the bladder wall. Nausea was observed in 1 case in which a large quantity of Tegafur was absorbed. No other side effects were noted.

[Thermotherapy for cancer of the urinary bladder in combination with tegafur suppository and picibanil--concentrations of tegafur in the serum and tissues in bladder carcinoma].

Thermotherapy combined with tegafur (FT-207) and Picibanil was performed in 13 patients with cancer of the urinary bladder. The thermotherapy was performed using a closed circulation type of thermotherapeutic apparatus manufactured for this experiment, by way of a 3-way catheter. The flow rate of the perfusate was 150 ml/min at a constant temperature of 43 degrees C. The temperature was monitored at the inlet and outlet catheter of the urethral meatus. The thermotherapy was performed for a 6-hour period once a week. A tegafur 3000-mg suppository was given 2 hours before the thermotherapy, and Picibanil was added to the perfusate at the time of thermotherapy. No anesthesia was used. Complete disappearance of the tumor was observed in 1 case. The concentrations of tegafur in the serum, tumor and bladder wall were measured simultaneously. The levels obtained at two hours after rectal administration of the tegafur 3000-mg suppository were 19.420 mcg/ml for FT-207 and 0.025 mcg/ml for 5-FU in the serum, 13.170 mcg/g for FT-207 and 0.140 mcg/g for 5-FU in the bladder tumor, and 20.447 mcg/g for FT-207 and 0.252 mcg/g for 5-FU in the bladder wall. Eruption was observed in 1 case. No other side effects were noted.

A study of the interaction between tRNASer and seryl-tRNA synthetase from bovine liver.

Study by chemical modification of Ser, Arg, His residues and sulfhydryl groups on bovine seryl-tRNA synthetase showed that Ser residues appeared to be unnecessary for the recognition mechanism, but Arg and His residues were essential. It was considered that different sulfhydryl groups related with each recognition of tRNA and ATP. Poly-arginine inhibited the interaction between serine tRNA and SerRS. The CD spectra of a mixture of serine tRNA and poly-arginine indicated that higher-order structure of tRNA changed. Furthermore, the Km and Vmax values of bovine serine isoacceptor, yeast serine tRNA and E. coli serine tRNA for bovine SerRS examined and it was discussed the differences of those base sequences.

The role of T cells in pathogenesis and protective immunity to murine malaria.

T-cell-mediated immunity to a virulent strain of Plasmodium berghei NK65 (Pb NK65) and to an attenuated derivative (Pb XAT) of the strain were examined in CBA mice by the administration of monoclonal antibodies against T-cell subsets or interferon-gamma (IFN-gamma). The injection of anti-CD8+ or anti-IFN-gamma delayed the mortality of mice infected with Pb NK65, although it did not affect the parasitaemia. In the late stage of PB NK65 infection, T cells, especially CD8+ T cells, were increased in number in the liver at the expense of splenic CD8+ T cells. These CD8+ T cells released IFN-gamma in culture without antigen stimulation and were thought to induce tumour necrosis factor-alpha (TNF-alpha) production by the cells in the liver. In mice infected with Pb XAT, or mice primed with Pb XAT and then challenged with Pb NK65, CD4+ T cells had a crucial role in preventing parasite growth and in protective immunity. IFN-gamma was again the key molecule in protective immunity. These results suggest that T cells stimulated with malaria antigen play important roles both in protective immunity and pathogenesis depending upon their subsets; CD8+ T cells in pathogenesis, and CD4+ T cells in protective immunity. These apparently contradictory responses may be mediated by the same cytokine, IFN-gamma.

A CXC chemokine receptor, CXCR5/BLR1, is a novel and specific coreceptor for human immunodeficiency virus type 2.

G protein-coupled receptors serve as coreceptors in the infection process of human immunodeficiency virus type-1 (HIV-1), type-2 (HIV-2), and simian immunodeficiency virus (SIV). In this study, we showed that a CXC-CKR, CXCR5/BLR1, is a novel coreceptor for HIV-2, but for neither HIV-1 nor SIV. The expression of CXCR5 was detected by polymerase chain reaction after reverse transcription of cellular mRNA from S+L-HOS/CD4 cells and MT-2 human T cells, and the CXCR5 gene was cloned into an expression vector. S+L-HOS/CD4 cells were susceptible to several HIV-2 strains but not most HIV-1 strains. To examine a coreceptor activity of CXCR5, we used NP-2/CD4, which is a human glioma cell line, NP-2, transduced with the CD4 gene that shows strict resistance to infection with HIV-1, HIV-2, SIVmac, SIVagm, or SIVmnd strain. When CXCR5 was transduced into NP-2/CD4 cells, they became highly susceptible to HIV-2ROD and HIV-2CBL23 strains in a CD4-dependent manner but to not to HIV-1 or SIV strains. Anti-CXCR5 monoclonal antibody and a ligand for CXCR5, BCA-1, inhibited HIV-2 infection to NP-2/CD4/CXCR5 cells. Our findings suggest a possibility that CXCR5/BLR1 serves as a coreceptor for HIV-2 strains in vivo.