Tumour necrosis factor α augments the inhibitory effects of CTLA-4-Ig on osteoclast generation from human monocytes via induction of CD80 expression.

Cytotoxic T lymphocyte antigen-4-immunoglobulin (CTLA-4-Ig) exerts anti-rheumatic action via negative regulation of the co-stimulation process between antigen-presenting cells and T cells. CTLA-4-Ig also binds to CD80/CD86 on monocytes of osteoclast precursors. However, little is known about the effect of CTLA-4-Ig on osteoclastogenesis in rheumatoid arthritis (RA). In this study we evaluated the effects of CTLA-4-Ig on osteoclast generation from human blood monocytes (PBM) and rheumatoid synovial fluid monocytes (RSFM). Highly purified monocytes were cultured with receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) in the presence of CTLA-4-Ig. CTLA-4-Ig inhibited RANKL-induced osteoclast generation in PBM and RSFM, as determined by tartrate-resistant acid phosphatase (TRAP) staining and bone resorption assay using osteo assay surface plates. In addition, CTLA-4-Ig reduced the gene and protein expressions of nuclear factor of activated T cells, cytoplasmic 1 (NFATc1) and cathepsin K during osteoclastogenesis. Furthermore, CTLA-4-Ig significantly inhibited cell proliferation during osteoclastogenesis. Interestingly, the gene expression of indoleamine 2,3-dioxygenase-1, an inducer of apoptosis, was enhanced by CTLA-4-Ig. We next examined the effect of tumour necrosis factor (TNF)-α, a major inflammatory cytokine in rheumatoid synovium, on the expression of CD80 and CD86 by flow cytometric analysis. TNF-α potently induced the surface expression of CD80, which is known to have much higher affinity to CTLA-4-Ig than CD86, and this induction was observed at mRNA levels. Interestingly, freshly prepared rheumatoid synovial monocytes also expressed CD80 as much as TNF-α-treated PBM. Furthermore, TNF-α enhanced CTLA-4-Ig-induced inhibition of osteoclastogenesis and cell proliferation. Taken together, the TNF-α-induced CD80 may augment CTLA-4-Ig-induced inhibition of osteoclastogenesis, suggesting that CTLA-4-Ig potently inhibits osteoclast differentiation and protects bone destruction in rheumatoid inflamed joints.

Specific downregulation of death-associated protein kinase enhances Fas-mediated apoptosis in the human differentiated endometrial adenocarcinoma cell line, HHUA.

PURPOSE OF INVESTIGATION: Death-associated protein kinase (DAPK) is a serine/threonine kinase that is well-known as a positive mediator of Fas-mediated apoptosis. Previous reports have shown that DAPK and Fas are expressed in human endometrial adenocarcinoma cells. In this study, we examined the effects of specific downregulation of DAPK expression on Fas-mediated apoptosis in the human endometrial adenocarcinoma cell line, HHUA. METHODS AND RESULTS: Transfection of DAPK small-interfering RNAs (siRNAs) into the HHUA cells reduced DAPK protein expression, and enhanced Fas-mediated apoptosis, in a dose-dependent manner. CONCLUSIONS: These results indicate that, in contrast to cases with other malignant tumor cells, DAPK negatively regulates Fas-mediated apoptosis in these human differentiated endometrial adenocarcinoma cells.

Strain dependency of B and T lymphoma development in immunoglobulin heavy chain enhancer (E mu)-myc transgenic mice.

The transgenic mice were produced by injecting eggs of B6 and C3H/HeJ mice with the human E mu-myc gene. Preferential development of B lymphomas was observed in the B6 transgenic mice, whereas the C3H/HeJ transgenic mice developed mostly T lymphomas. The phenotypic activation of B lineage cells but not of T lineage cells was detected in the prelymphomatous transgenic mice of both strains. The transgene was similarly expressed in B and T cells of the transgenic mice of both strains. These results suggest that a high incidence of T lymphomas in the C3H/HeJ transgenic mice may not be due to the preferential activation of or the preferential E mu-myc expression in T lymphocytes. When the bone marrow or fetal liver cells from the prelymphomatous transgenic mice of both strains were transferred into irradiated normal C3H/HeJ mice, most of the recipients developed T lymphomas. Moreover, even when irradiated B6 mice received the hematopoietic stem cells from the prelymphomatous B6 transgenic mice, the incidence of T lymphoma increased up to 50%. These findings suggest that B6 and C3H/HeJ mice might provide the environment that supports the development or growth of B and T lymphomas, respectively, and that such an environment could be modified by irradiation of the mice.

Large-vessel vasculitis associated with PEGylated granulocyte-colony stimulating factor.

A 71-year-old female with advanced endometrial cancer was treated with pegfilgrastim. She developed a fever within seven days, and contrast-enhanced computed tomography scans repeated within three days revealed rapidly progressive thickening of the aortic wall. When clinicians administer PEGylated granulocyte-colony stimulating factor (G-CSF) to cancer patients, drug-associated vasculitis should be suspected. This report discusses the manifestation of G-CSF-associated large-vessel vasculitis (LVV).

Radiation reduces carboplatin sensitivity and enhances nedaplatin sensitivity in cervical squamous cell carcinoma in vitro.

BACKGROUND: The study was performed to examine how the platinum anticancer drugs other than cisplatin, such as carboplatin (CBDCA) and nedaplatin (NEP) can be effectively used in chemoradiotherapy for cervical squamous cell carcinoma patients. MATERIALS AND METHODS: The radiosensitive human cervical squamous cell carcinoma cell line ME180 was examined to investigate the radiation effects on CBDCA and NEP sensitivities of the cells. RESULTS: Irradiation significantly reduced cellular CBDCA sensitivity. There were no significant changes in CBDCA sensitivity between the cells concurrently irradiated and those treated with CBDCA 8 h before or 8 h after irradiation. However NEP sensitivity of the cells treated 8 h before or 8 h after irradiation was significantly higher than that in cells concurrently irradiated. CONCLUSIONS: Although CBDCA sensitivity in the concurrently irradiated cells is reduced, NEP sensitivity is enhanced by irradiation. NEP, but not CBDCA, therefore, may be a candidate anticancer drug for concurrent chemoradiotherapy for cervical cancer. For the greatest efficacy, NEP should be administered to patients several hours before or after irradiation.

Irradiation reduces bleomycin sensitivity in cervical squamous cancer cells in vitro.

PURPOSE OF INVESTIGATION: The study was performed to examine how bleomycin (BLM) and peplomycin (PEM) should be effectively used in radiotherapy for cervical squamous cancer patients. METHODS: The effects of BLM on radiosensitivity and the effects of radiation on the sensitivity to BLM of cancer cells were investigated using the radiosensitive human cervical squamous cell carcinoma cell line ME180. RESULTS: BLM treatment did not affect radiosensitivity. However, irradiation significantly reduced cell BLM sensitivity in a dose-dependent manner. There was no significant difference in BLM sensitivity and PEM sensitivity between cells concurrently irradiated and those treated with BLM or PEM 8 h before or 8 h after irradiation. CONCLUSION: Since sensitivity to BLM is reduced during irradiation, BLM should be administered to cervical cancer patients as an adjuvant chemotherapeutic drug after completion of radiotherapy.

Effective chemoradiotherapy protocol with 5-fluorouracil for cervical squamous cell carcinoma in vitro.

PURPOSE OF INVESTIGATION: 5-Fluorouracil (5FU) is frequently used in concurrent chemoradiotherapy for patients with advanced cervical cancer, although its optimal chemoradiotherapy protocol has not yet been established. In search of an optimal chemoradiotherapy protocol, some in vitro experiments were carried out. METHODS: The radiosensitive human cervical squamous cell carcinoma cell line ME180 was examined to investigate the effects of 5FU on radiosensitivity and the effects of irradiation on 5FU-sensitivity. RESULTS: 5FU dose-dependently enhanced cellular radiosensitivity at therapeutic concentrations. Although high doses of y-ray irradiation significantly reduced the 5FU-sensitivity, a low dose of irradiation at therapeutic doses (< 2.5 Gy) had no effect on 5FU-sensitivity of the irradiated cells. Cells pretreated with 5FU eight hours before irradiation showed significantly higher 5FU-sensitivity than cells concurrently treated with 5FU and irradiation. In contrast, cells treated with 5FU eight hours after irradiation showed significantly lower 5FU-sensitivity than cells concurrently treated with 5FU and irradiation. Moreover, all four post-irradiation surviving subclones obtained from repeatedly irradiated ME180 cells showed significantly lower 5FU-sensitivity than the non-irradiated parent cells. CONCLUSION: 5FU acts as a radiosensitizer for cervical squamous cell carcinoma and 5FU-sensitivity is reduced in irradiated cells. Therefore, 5FU administration immediately before irradiation may be a more effective treatment than concurrent chemoradiotherapy or post-irradiation chemotherapy with 5FU.

Radiation enhances cisplatin-sensitivity in human cervical squamous cancer cells in vitro.

UNLABELLED: PURPOSE AND METHODS OF INVESTIGATION: Cisplatin (CDDP) is regularly used in concurrent chemoradiotherapy in patients with advanced cervical cancer although an effective protocol of chemoradiotherapy with CDDP has not yet been established. In search of a better chemoradiotherapy protocol, we investigated both CDDP effects on radiosensitivity and irradiation effects on CDDP-sensitivity using the radiosensitive human cervical squamous cell carcinoma cell line ME180. RESULTS: We found that CDDP did not affect cellular radiosensitivity, and that irradiation significantly enhanced CDDP-sensitivity. Moreover, all the four post-irradiation surviving subclones obtained from repetitively irradiated ME180 cells showed significantly higher CDDP sensitivities than those of the non-irradiated parent cells. CONCLUSION: These results suggest that an effective protocol would involve the concurrent administration of CDDP with radiotherapy and further administration following completion of radiotherapy in order to achieve higher CDDP-sensitivities.

Epoc-1: a POU-domain gene expressed in murine epidermal basal cells and thymic stromal cells.

POU-domain transcription factors are known as developmental regulators which control organ development and cell phenotypes. In order to clarify the roles of POU-domain transcription factors in cell differentiation, we cloned a novel POU family gene, Epoc-1, from a murine thymus cDNA library. The amino acid (aa) sequence of the POU-specific domain of Epoc-1 is almost identical to those of Oct-1 and Oct-2. However, within the POU-homeodomain, 13 out of 60 aa differ between Epoc-1 and Oct-2. Recombinant Epoc-1 products were found to bind specifically to the octamer sequence. Epoc-1 was found to be expressed in skin, thymus, stomach and testis. In situ hybridization experiments and RNase protection assays indicated that Epoc-1 is expressed in the epidermal basal cells of the skin, which contain stem cells unipotent for keratinocyte differentiation and in thymic stromal elements. These results suggest that Epoc-1 might be one of the developmental regulators which controls epidermal development and thymic organogenesis.

Regulation of transcription factor C/ATF by the cAMP signal activation in hippocampal neurons, and molecular interaction of C/ATF with signal integrator CBP/p300.

The CCAAT/enhancer binding proteins related activating transcription factor, C/ATF, is a mouse leucine-zipper transcription factor which is structurally homologous to ApCREB2, a suppressor integral to long-term synaptic plasticity in Aplysia. To gain a clue to whether C/ATF is involved in long-term plasticities of brain, we examined if the expression levels of C/ATF are modulated by cAMP, an inducer crucial for memory formation in Aplysia, Drosophila and mice. Our in situ hybridization analysis revealed the expression of C/ATF mRNA in hippocampal neurons. C/ATF protein levels increased after the cAMP signal stimulation in hippocampal neurons, while C/ATF mRNA levels remained constant. The human activating transcription factor 4 (hATF4), another homolog of ApCREB2, interacts with multiple domains of the coactivator CREB-binding protein (CBP), resulting in the potentiation of its ability to activate transcription. As expected, C/ATF was found to interact with three domains of CBP including CREB binding domain or kinase-inducible interaction (KIX) domain, the third cysteine-histidine-rich region (CH3 domain) and the nuclear receptor coactivator p160/SRC-1-interacting domain. Interestingly, C/ATF was further found to interact strongly with CREB binding protein/p300 (CBP/p300) CH1 domain. Mammalian two hybrid assays indicated that the interaction between C/ATF and CBP/p300 can occur in mammalian cells, and that the p300 CH1 domain is critical for the interaction. Thus, C/ATF may be implicated in transcription-dependent phase of hippocampal long-term plasticities through the modulation of its protein level under cAMP signal and the interaction with signal integrator, CBP/p300.

Hypotensive effects of an angiotensin II type 1 receptor antagonist differ between exercised and sedentary rats aged from 4 to 19 weeks.

The hypotensive effects of an angiotensin II type 1 receptor antagonist TCV-116 were accelerated by swimming training in the early period of chronic administration in young rats. In the latter period, however, a severe decrease in blood pressure that appeared during long-term administration of TCV-116 was prevented by swimming training. Therefore exercise may contribute to the stabilization of blood pressure in young rats treated with TCV-116.

Factors contributing to phoneme recognition ability of users of the 22-channel cochlear implant system.

A study was carried out to determine which factors contributed to the vowel and consonant recognition ability of recipients of the 22-channel cochlear implant system. On the basis of the statistical analysis, no isolated factor showed a strong correlation with vowel recognition score. On the other hand, negative correlations were found between patients' consonant recognition scores and postoperative psychophysical percepts such as threshold levels and maximum comfortable loudness levels. However, multiple regression analysis also showed that the combination of lower threshold levels, a larger number of usable electrodes, and wider dynamic ranges contributed to higher consonant recognition scores.

Clinical features of paroxysmal positional vertigo presenting combined lesions.

Benign paroxysmal positional vertigo (BPPV) is one of the common vestibular disorders. Canalolithiasis is thought to be a likely lesion. A canalith repositioning procedure (CRP by Epley) generally yields good resolution of vertigo and nystagmus. The authors confirmed the efficacy of this procedure on typical BPPV of the posterior semicircular canal type. We designed a new procedure for BPPV of the lateral canal type, which also yielded satisfactory results. BPPV sometimes presents a nystagmus pattern, which suggests multiple lesions. We have seen eight cases of BPPV showing nystagmus that combines both the posterior and the lateral canal types. Combined CRP was performed on these cases, which again gave good clinical results. Other BPPV cases were associated with central lesions. We must be aware that BPPV may involve multiple canals and may be associated with central lesions.

Japanese-language results for the speak strategy from Cochlear Pty Limited.

Recognition of speech sounds with a new speech-coding strategy, Speak, was compared with that of the Multipeak (Mpeak) strategy. Results of consonant, word, and sentence tests on 10 postlingually deaf subjects demonstrated that the performance of the Speak strategy significantly exceeded that of the Mpeak strategy under both quiet and noise conditions. Information transfer ratios for consonant features were improved by 20% to 40% with the Speak strategy under the noise condition. Thus, the Speak coding strategy can provide better speech sound perception than the Mpeak coding strategy.

Speech perception with multi-channel cochlear implant of short duration pulse strategy.

A multi-channel cochlear implant (Nucleus type) was implanted in a 40-year-old female Japanese patient who became totally deaf after meningitis. The formant-based speech processing strategy was used, but a narrow pulse width of 22-42 microseconds was required because of intermittent difficulty in controlling the pulse amplitude. The patient was tested with a speech tracking test and could recognize 24 bunsetsues (the minimum meaningful unit of the Japanese sentence) per minute using the cochlear implant plus lipreading and 14.3 bunsetsues for the lipreading alone after 3.5 months' training. The patient was also able to understand usual conversational sentences spoken a little slowly. Scores of vowel and consonant tests reached 70% and 54% respectively for the cochlear implant alone, and 100% and 73% for the cochlear implant plus lipreading. This study has also shown that cochlear stimulation with very narrow pulse widths can be used, and restore speech comprehension ability for the Japanese.

[Validity of various obesity indices calculated from height and weight in adult females using the underwater-weighing method as a reference].

We evaluated association between excess percent body fat (%Fat) and various obesity indices calculated from height and weight in 322 adult females. %Fat was measured by the underwater-weighing method, and obesity indices were based on the following 5 methods; Broca-Katsura method (Katsura method), Kato-Wataya method (Kato method), Japan Society for the Study of Obesity method (BMI method; based on the body weight at which BMI is 22), Meiji Life Insurance Co. method, and Table and Figure for the Assessment of Obesity and Leanness by the Ministry of Health and Welfare (MHW method). %Fat was 30% or more (obese) in 73 females (22.7%), 25-30% in 97 (30.1%), 20-25% in 88 (27.3%), and less than 20% in 64 (19.9%). The correlation coefficient between the obesity indices and %Fat were 0.71 for the Katsura method, 0.70 for the Kato-method, 0.72 for the BMI method, 0.70 for the Meiji Life Insurance Co. Method, and 0.63 for the MHW method, being significant for all methods (P < 0.01). When the cut-off point was set as 110% for each obesity index, sensitivity was the highest for the Katsura method (67.1%), and specificity was the highest for the Meiji Life Insurance Co. method (95.2%). Receiver operating characteristic (ROC) curves were similar in figure for the Katsura method, Kato-method, BMI method, and Meiji Life Insurance Co. method. For the MHW method, however, the curve was slightly farther from the point of sensitivity of 100% and 1-specificity of 0% than the others. Excess fat accumulation could not be accurately assessed by the obesity indices calculated from body height and weight. Validity was similar among the obesity indices except for the MHW method.

[Body composition (densitometry-hydrostatics), skinfold thickness, BMI and their relationships in junior high school girls].

A study was conducted to evaluate body composition (Hydrostatics = Under-Water Weighing), skinfold thickness, and BMI in 97 junior high school girls, aged 12 to 15, in Nagasaki City. Body density was measured by under-water weighing, and the two compartments--fat weight (Fat) and lean body mass (LBM, or fat free weight: FFW)--were calculated using the formulas of Brozek et al. The results were as follows; 1. Mean values of body density were 1.0501 in first grade girls, 1.0476 in the second grade, and 1.0466 in the third grade. 2. Mean values of percentage body fat (% Fat) were 21.1% in the first grade, 22.1% in the second and 22.5% in the third. 3. Lean body mass (LBM) and LBM/Height increased significantly with advance from the first to the third grade. 4. The correlation coefficient between percent body fat and the sum of two skinfold thicknesses, the sum of three skinfold thicknesses, or the sum of seven skinfold thicknesses were 0.81, 0.80, and 0.82 respectively and all statistically significant (p < 0.001). 5. The correlation coefficient between BMI and the sum of two skinfold thicknesses, the sum of three skinfold thicknesses (r = 0.841) or the sum of seven skinfold thicknesses were 0.85, 0.84, and 0.84 respectively, and all statistically significant (p < 0.001). 6. Mean values of BMI, WSR, ASR, and WHR in all subjects (n = 97) were 19.6, 0.72, 0.39 and 0.43 respectively.

[Hydrostatic weighing, skinfold thickness, body mass index relationships in high school girls].

A study was conducted to evaluate body composition by hydrostatic weighing, skinfold thickness, and body mass index (BMI) in 102 senior high school girls, aged 15 to 18 in Nagasaki City. Body density measured by the underwater weighing method, was used to determine the fat weight (Fat) and lean body mass (LBM. or fat free weight: FFW) utilizing the formulas by Brozek et al. The results were as follows; 1. Mean values of body density were 1.04428 in the first grade girls, 1.04182 in the second grade, and 1.04185 in the third grade. 2. Mean values of percentage body fat (%Fat) were 23.5% in the first grade, 24.5% in the second and 24.5% in the third. 3. Percentage body fat (%Fat), lean body mass (LBM) and LBM/Height were not significantly with different advance of grade from the first to the third. 4. The correlation coefficients between percent body fat and the sum of two skinfold thicknesses, the sum of three skinfold thicknesses and the sum of seven skinfold thicknesses was 0.78, 0.79, and 0.80 respectively and were all statistically significant (p < 0.001). 5. The correlation coefficients between BMI and the sum of two skinfold thicknesses, the sum of three skinfold thicknesses and the sum of seven skinfold thicknesses was 0.74, 0.74, and 0.74 respectively and were all statistically significant (p < 0.001). 6. Mean values of BMI, Rohrer index and waist-hip ratio (WHR) in all subjects (n = 102) were 20.3, 128.2 and 0.72 respectively.

[Body density assessment utilizing skinfold thickness and age in Japanese women].

A study was conducted to investigate the validity of skinfold-based prediction equations for body density (g/ml) by Nagamine and Suzuki (1964), and to formulate more convenient and more useful equations for predicting body density from skinfold and age in women. Subjects of the study were 512 healthy women aged 18-66 years in or near Nagasaki City. The dependent variable in the multiple regression equation, body density, was determined by hydrostatic weighing. Independent variables included eight skinfolds, the sum of two skinfolds (triceps, subscapular), the sum of three skinfolds (triceps, subscapular, and abdominal), age, and body surface area. Skinfolds were measured with an Eiken-model skinfold caliper. Age (mean 30.1, range 18-66 yrs.), weight (mean 52.6, range 38.0-83.3 kg), height (mean 157.0, range 142.0-172.0 cm), and body density, (mean 1.04125, range 0.98806-1.08650 g/ml) were also recorded. Percent body fat was calculated using the formula by Brozek et al. and ranged from 6.4% to 48.3%. Multiple correlation coefficients (MR) and standard error (SE) of 10 regression equations (A-J) for predicting body density in women were compared. The best-fitting and the most convenient prediction equation for body density was equation-E. The regression equation developed for predicting body density was: body density = 1.07931-0.00059 x sum of three skinfolds(mm)-0.00015 x age (MR = 0.77 and SE = 0.0089). The equation was cross-validated on a different sample of 46 women. The correlation coefficient between predicted and hydrostatically determined body density was 0.813 (p < 0.001). Equation-E (Tahara's equation) appears to be useful in body density analysis particularly when the subjects are Japanese women, aged 18-50 yrs, with percent body fat 17 to 34%.

[Body density assessment utilizing skinfold thickness and age in Japanese men].

A study was conducted to investigate the validity of skinfold-based prediction equations for body density (g/ml) by Nagamine and Suzuki (1964), and to formulate more convenient and more useful equations for predicting body density from skinfold and age in men. Subjects of the study were 257 healthy men aged 19-60 years in or near Nagasaki City. The regression equation for the dependent variable, body density, was determined by hydrostatic weighing. Independent variables included eight skinfolds, the sum of two skinfolds (triceps, subscapular), the sum of three skinfolds (triceps, subscapular, and abdominal), age, and body surface area. Skinfolds were measured with an Eiken-model skinfold caliper. Age (mean 33.1, range 19-60 yrs.), weight (mean 65.3, range 46.6-107.7 kg), height (mean 168.8, range 152.3-185.4 cm), and body density (mean 1.05874, range 1.00860-1.09020 g/ml) were also recorded. Percent body fat was calculated using the formula by Brozek et al. and ranged from 6.1% to 38.9%. Multiple correlation coefficients (MR) and standard error (SE) of 10 regression equations (A-J) for predicting body density in men were obtained. The best-fitting and the most convenient prediction equation for body density was equation-E.: body density = 1.09556-0.00062 x sum of three skinfolds (mm)-0.00028 x age (MR = 0.815 and SE = 0.0087 g/ml). The equation was cross-validated on a different sample of 45 men. The correlation coefficient between predicted and hydrostatically determined body density was 0.781 (p < 0.001). Equation-E (Tahara's equation) appears to be useful in body density analysis particularly when the subjects are Japanese men, aged 18-50 yrs, with percent body fat 10 to 30%.