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BackgroundPatients with rheumatoid arthritis (RA) who have sarcopenia and stiff or painful joints might be at increased risk of falls and fractures.ObjectivesThe present study aimed to prospectively identify the incidence of clinical fractures and associated risk factors in patients with RA in a cohort study named the TOMORROW (UMIN000003876) that started in 2010.MethodsWe evaluated anthropometric parameters, bone mineral density (BMD), disease activity, RA medication at entry and observed the incidence of clinical fractures during ten years in 202 patients with RA (mean age, 58.6 y; medication with biological agents, 54.9%) and 202 age- and sex-matched non-RA volunteers (Vo) (mean age, 57.4 y). We compared the incidence of clinical fractures between patients with RA and Vo for ten years, and analyzed the risk factors for clinical fractures using Cox proportional hazard model.ResultsThe incidences of clinical fractures were 0.036 and 0.024/person-year in patients with RA and Vo, respectively. Cox proportional hazard model revealed that low BMD at the thoracic vertebrae (< 0.7 g/cm2) and history of fractures at entry were significantly associated with the incidence of clinical fractures (Hazard ratio [HR]1.737, p=0.020 and HR1.514, p=0.047, respectively) in all participants. RA morbidity, however, was not (HR1.398, p=0.112). In patients with RA, medication with GC at entry was a significant risk factor for clinical fractures (HR1.898, p=0.017). Additionally, a mean GC dose (≥ 2 mg/day) at entry and during the ten-year period increased risk for fractures (HR 2.189, p=0.004, 1.866, p=0.022, respectively).ConclusionRA per se was not a risk factor for clinical fractures in this cohort. Low BMD at the thoracic vertebrae at entry and the use of GC with even low dose at entry and during ten years were significantly associated with an increased frequency of clinical fractures among patients with RA.Disclosure of InterestsNone declared

BackgroundPatients with rheumatoid arthritis (RA) who have muscle weakness and stiff or painful joints might be at increased risk of falls and fractures.ObjectivesThe present study prospectively investigates correlations between decreasing doses of glucocorticoid (GC) and the incidence of clinical fractures in patients with RA based on the ten-year findings of the TOMORROW study (UMIN000003876) that started in 2010.MethodsWe evaluated anthropometric parameters, bone mineral density, disease activity, RA medication, and the incidence of clinical fractures over a period of ten years in 202 patients with RA (mean age, 58.6 years; mean disease duration, 14.0 years). We also investigated the effects of GC doses on the incidence of clinical fractures over the same period in patients with RA using multivariate regression analysis.ResultsThe incidence of clinical fractures for ten years in patients with RA was 0.036/person-year. There were 89 patients (44.1%) treated with GC at least once during ten years. The incidences of clinical fractures in patients with RA treated with and without GC during ten years were 0.052 and 0.026/person-year, respectively. After adjusting for fracture risk factors including age, sex, smoking, and body mass index, cox proportional hazard model revealed that GC dose of ≥ 2 mg/day at baseline was a significant risk factor for clinical fractures (Hazard ratio [HR]:2.430; 95%CI, 1.040-5.675, p=0.040). Although the risk for clinical fractures did not decrease by just reducing the dose of GC (HR:4.505; 95%CI, 0.589-34.457, p=0.147), it was significantly lower if the dose of GC could be reduced to zero during ten years (HR:0.407; 95%CI, 0.194-0.857, p=0.018).ConclusionMedication with even low dose of GC are apparently significantly associated with an increased frequency of clinical fractures among patients with RA. However, if the dose of GC was reduced to free during ten years, the clinical fracture risk could become lower. We concluded that we should decrease the dose of GC to free after controlling disease activity of RA.Disclosure of InterestsNone declared

BackgroundTo investigate if FRAX in patients with RA can predict the incidence of new clinical fractures for 10 years by using the 10-year data of the TOMORROW study (UMIN000003876) which is a prospective cohort study.ObjectivesTo investigate if FRAX in patients with RA can predict the incidence of new clinical fractures for 10 years by using the 10-year data of the TOMORROW study (UMIN000003876) which is a prospective cohort study.MethodsWe calculated ten-year probability of major osteoporotic fracture (FRAX) in 208 RA patients and 205 sex- and age-matched volunteers (Vo), and compared FRAX with the incidence of clinical fractures for 10 years.ResultsThe mean FRAX were 14.5 and 8.8% in 175 RA patients and 168 Vo, respectively, in whom we could calculate FRAX at baseline and complete to investigate the incidence of clinical fractures for 10 years from baseline. The mean FRAX in RA patients was significantly higher than that in Vo (PConclusionFRAX and the incidence of clinical fractures for 10 years were significantly higher in RA patients than them of Vo. We confirmed that FRAX was the risk factor for clinical fractures in actual clinical practice.Disclosure of InterestsNone declared

BackgroundRomosozumab, a monoclonal antibody that binds sclerostin, increases bone formation and decreases bone resorption. Romosozumab has an excellent therapeutic effect on patients with osteoporosis. However, there are few reports investigated the efficacy and the effect of various factors relative to osteoporosis in real-world.ObjectivesWe evaluated bone mineral density (BMD) and bone metabolism marker (BMM) change in osteoporosis patients treated with romosozumab, and assessed the effect of various factors, such as preosteoporosis treatment, history of fragility fracture and combination of vitamin D.MethodsThis study included 141 osteoporosis patients (132 female, mean age: 78.8 ± 7.5 years, mean hight: 151.1 ± 7.5 cm, mean weight: 50.9 ± 8.9 kg) treated with romosozumab. BMD measurements using dual energy X-ray absorptiometry (DXA) and BMM measurements using N-terminal propeptide of type 1 collagen (P1NP) were evaluated at 0, 4, 6, and 12 months after treatment. DXA were performed at the lumbar spine (n=92), at proximal femoral and femoral neck (n=92), and at distal third radius (n=49). We evaluated the influence of preosteoporosis treatment, history of fragility fracture and combination of vitamin D for BMD change.ResultsBMD at lumbar spine (5.2%: pConclusionRomosozumab improved BMD at lumbar and femoral independently regardless of preosteoporosis treatment, history of fragility fracture and combination of vitamin D.References[1]Felicia Cosman et al. Romosozumab Treatment in Postmenopausal Women with Osteoporosis. N Engl J Med 2016; 375:1532-1543, DOI: 10.1056/NEJMoa1607948Disclosure of InterestsNone declared

Dielectric barrier discharge (DBD) treatment is one of the methods used to make carbon nanofibers stand up on substrates. Upright carbon nanofibers are used as field emission materials. We have used twisted carbon nanofibers (CNTws) as field emission materials and treated printed CNTws on substrates using DBD. In this report, we examine the effects of DBD conditions on the uprightness of the CNTws. The DBD experimental parameters were as follows: 1) N2/He gas mixture ratio, 2) pulse frequency, and 3) Pt coating on the CNTw surface. The lengths of upright CNTws from the substrate surface and from the surface of a printed CNTw dot were measured using scanning electron microscopy. N2 gas was shown to be crucial for generating streamer discharges and making the CNTw stand up on the substrate. As the pulse frequency increased, the lengths of the upright CNTws and their number density increased. This is explained by an increase in the number of streamers; the streamers move about over the substrate surface. Pt coating lowered the onset voltage for field emission from the CNTws although the number of upright CNTws was less than that without the Pt coating.

Objectives. Dysbaric osteonecrosis (DON) is a complication of ineffective decompression following exposure to high-pressure environments. This study was designed to determine risk factors for the occurrence of DON in divers. Methods. Fifty-six male divers received skeletal examinations by radiography to assess the occurrence of DON. A questionnaire was used to obtain clinical and diving information, including diving experience and maximum diving depth. Blood samples were collected to analyse the levels of plasminogen activator inhibitor (PAI)-1, cholesterol, triglyceride, lowdensity lipoprotein, very low-density lipoprotein, high-density lipoprotein, apolipoprotein A1 and apolipoprotein B. Results. Lesions of DON were detected in 31 of the 56 (55%) divers. Multivariate logistic regression analysis showed that high levels of PAI-1, a coagulation marker (odds ratio 4.281; P^0.0296) and great maximum diving depth (odds ratio 5.627; P^0.0231) were independent predictors of DON. Conclusions. This study has shown the presence of coagulation abnormality in divers with DON. This result suggests that a pharmacological approach incorporating the use of an anticoagulant may represent a potential strategy for the prevention of DON.

Our study describes the mid-term clinical results of the use of transtrochanteric valgus osteotomy (TVO) for the treatment of osteoarthritis of the hip secondary to acetabular dysplasia. The operation included valgus displacement at the level of the lesser trochanter, and lateral displacement of the greater trochanter by inserting a wedge of bone. We reviewed 70 hips. The mean age of the patients at operation was 44 years (14 to 59). Most (90%) had advanced osteoarthritis. The scores for pain and gait had improved significantly at a mean follow-up of 9.4 years. The rate of survival until an endpoint of a further operation during a follow-up of ten years was 82%. The survival rate was 95% in patients with unilateral involvement who were less than 50 years of age at operation. TVO is a useful form of treatment for advanced osteoarthritis of the hip, particularly in young patients with unilateral disease.

We have studied the correlation between the prevention of progressive collapse and the ratio of the intact articular surface of the femoral head, after transtrochanteric rotational osteotomy for osteonecrosis. We used probit analysis on 125 hips in order to assess the ratio necessary to prevent progressive radiological collapse over a ten-year period. The results show that a minimum postoperative intact ratio of 34% was required. This critical ratio may be useful for surgical planning and in assessing the natural history of the condition.

An intramolecular anhydride formation in 5-isopropenyl-4,5-dihydrofuran-2,3-dicarboxylic acid caused a new ring expansion to give 4,7-dihydro-6-methyloxepin-2,3-dicarboxylic anhydride (2b). An aminolysis of 2b with benzylamine gave a mixture of two isomeric carboxamides, N-benzyl-4,7-dihydro-6-methyloxepin-2-carboxamide and trans-1-(benzyloxamoyl)-2-isopropenylcyclopropane. Also, the ring expansion and ring contraction are discussed.

Diethyl 5-isopropenyl-4,5-dihydrofuran-2,3-dicarboxylate 1a and methyl 5-isopropenyl-3-methoxy-carbonyl-4,5-dihydrofuran-2-acetate 2a were prepared by cylization of diethyl 2-oxosuccinate or dimethyl 3-oxoglutarate with (E)-1,4-dibromo-2-methyl-2-butene. Their chemical properties were studied.

Not every postmenopausal woman with a low level of estrogen suffers from osteoporosis, and no correlation of bone density with serum estrogen level, but a significant correlation with adrenal androgens is often noted. Vitamin D3 has been reported to be osteoclastic in vitro, whereas the effectiveness of vitamin D3 for the treatment of osteoporosis is clinically relevant. To study the roles of these factors in the development of osteoporosis, we characterized aromatase activity converting androgens to estrogens in human osteoblasts, because postmenopausal women maintain considerable levels of adrenal androgens. Glucocorticoids at 10(-9)-10(-7) M transiently induced the expression and enzymatic activity of aromatase cytochrome P450 (P450AROM) in primary cultured osteoblasts, and the Km value for androstenedione (4.7 +/- 2.9 nM) was lower than that in adipose tissue and skin. Human osteoblasts showed a promoter specificity different from that found in other tissues. 1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] alone did not induce aromatase activity, but enhanced and maintained glucocorticoid-induced P450AROM gene expression. This synergistic effect was not observed by other sex steroids or retinoic acids. The enhancement of P450AROM activity by 1,25-(OH)2D3 varied from 0.94-fold (no enhancement) to 2.40-fold (maximal enhancement) among the individual human osteoblasts examined, but the magnitude of the enhancement was significantly correlated with the level of vitamin D receptor messenger RNA (P0.05). Cycloheximide did not abolish the synergistic effect of 1,25-(OH)2D3, suggesting that de novo protein synthesis is not required for the synergism with 1,25-(OH)2D3. These results suggest that bone tissue can synthesize estrogen from adrenal androgens by a unique aromatase activity depending on the level of vitamin D receptor expressed.

A multimeric peptide, Ac-Y16, consisting of 16 YIGSR sequences from laminin was evaluated for its effect on experimental metastasis, angiogenesis and tumour growth of HT1080 human fibrosarcoma cells. Co-injection of 0.5 mg per mouse of Ac-Y16 i.v. with HT 1080 cells inhibited lung colonisation by 100%, whereas 0.5 mg per mouse of monomeric Ac-YIGSR-NH2(AcY1) inhibited by 94%. Ac-Y16 did not show any direct cytotoxicity in tumour cells in vivo. The effect of the peptides on angiogenesis and tumour growth respectively were evaluated by counting areas of neovessels and weighing tumours after the s.c. implantation of HT1080 cells with basement membrane extracts and the peptide into nude mice. Co-injection of 0.5 mg per mouse of AC-Y16 s.c. with HT1080 cells inhibited angiogenesis and tumour growth by 92% (P0.05) respectively. It can be inferred from these data that anti-tumour effects of Ac-Y16 are likely to result from anti-angiogenesis. Intraperitoneal administration of Ac-Y16 was also effective in inhibiting angiogenesis, tumour growth and lung colonisation of HT1080 cells. It was concluded that the multimeric YIGSR-containing peptide, Ac-Y16, inhibits angiogenesis, tumour growth and experimental metastasis more than the monomeric form and that it is active when administered i.p., iv. and s.c. Images Figure 3

Diethyl 5-isopropenyl-4,5-dihydrofuran-2,3-dicarboxylate 1a and methyl 5-isopropenyl-3-methoxy-carbonyl-4,5-dihydrofuran-2-acetate 2a were prepared by cylization of diethyl 2-oxosuccinate or dimethyl 3-oxoglutarate with (E)-1,4-dibromo-2-methyl-2-butene. Their chemical properties were studied.

An intramolecular anhydride formation in 5-isopropenyl-4,5-dihydrofuran-2,3-dicarboxylic acid caused a new ring expansion to give 4,7-dihydro-6-methyloxepin-2,3-dicarboxylic anhydride (2b). An aminolysis of 2b with benzylamine gave a mixture of two isomeric carboxamides, N-benzyl-4,7-dihydro-6-methyloxepin-2-carboxamide and trans-1-(benzyloxamoyl)-2-isopropenylcyclopropane. Also, the ring expansion and ring contraction are discussed.

A program of autologous blood transfusion in 101 patients who underwent spinal surgery was analyzed. Autologous transfusion was achieved by intraoperative blood salvage using a cell saver and with predeposited blood. The techniques of predeposition of blood included freezing the blood and using a storage solution. Forty-eight patients had scoliosis and received mainly cryopreserved autologous blood and 53 patients had other spinal diseases. Surgery was performed using only autologous blood transfusion in more than 90% of all patients cases. The program was well tolerated by the patients and easily managed by the blood center staff. There were no severe complications associated with this program. The use of instrumentation and extensive spinal fusion were found to be factors associated with increased intraoperative blood loss. This finding suggests that instrumentation surgery and extensive spinal fusion are indications for autologous blood transfusion. Cryopreserved autologous blood transfusion is an effective method for storing a sufficient volume of blood for scoliosis surgery without affecting the patients' preoperative hemodynamic status.

Several difficult problems arise in the treatment of necrotic bone disorders such as femoral head necrosis or Kienböck's disease for which no definitive treatment as yet exists. The recent literature has recommended vascularized bone transfers as suitable for these disorders, but there have been few experimental studies done and resulting effects are still currently obscure. Using a rabbit model, the effects of vascularized bone graft on the surrounding necrotic bone were analyzed. Results suggest that the periosteal muscle cuff remains as fibrous or fatty tissue and interferes with the bony union between the graft and the host bone; the graft was found to be gradually resorbed. The main function of the procedure appears to be the provision of vessels into avascular bone.

The development of the thermal stress free waterproof pan for prefabricated bathroom, thermo floor was described in this paper. For development, the evaluation methods of cold sensitive index was discussed and defined as an index of discomfort when feet touched with surface of the pan. Ceramic balloon filler was used for reducing the thermal conductivity of the material. As a physiological evaluation, the change of temperature of feet bottom and blood pressure in touching thermo floor were confirmed to be lower than in touching existing pan. So, thermo floor is effective for decreasing the thermal stress

We proposed a new concept of medical care system, named the "hyper hospital" which is constructed in computer based electronic information network. It is built as a distributed system on the network and consists of all kinds of conventional medical care facilities in both the real and the imaginary spaces. The virtual space of the hyper hospital and private information are owned and exclusively controlled by the patient, thus the maximum protection of the privacy of the patients can be observed. In the hyper hospital, an innovative human interface must be required for the consultation, treatment and other communication to the medical staff. This is especially true if the users are of severely disabled patients. The purpose of the present study is to establish a new human interface based on the event related potential, which enables those kind of patients to communicate with others including medical care staffs without any physical actions. >

In Fukuoka Prefecture, in south-western Japan, a regional screening program for osteoporosis was conducted from 1994 to 1995. The screening level in the bone mineral density (BMD) at the distal non-dominant radius was equal to or less than two standard deviations below age-specific mean (or = -2.0 SD). In 1177 examinees with natural menopause (mean age: 61.4, range: 42-88), 56 of those who were screened were subsequently radiologically confirmed by orthopedic specialists to have osteoporosis (case group). They were then compared with 802 normal BMD (or =-1.0 SD) women (reference group) with their lifestyle and reproductive characteristics. The adjusted odds ratio (OR) and its 95% confidence interval (CI) were calculated using a logistic regression model. A significant increase in the ORs for osteoporosis based on the number of years since menopause was observed for 7-13 years since menopause (OR = 2.3; 95% CI: 1.0-5.4) compared with7 years, however, no increasing trend in risk was evident in 14+ years since menopause (OR = 1.4; 95% CI: 0.4-5.1). Thus, the elevated risk continued up to around 10 years since menopause. These findings are consistent with previous studies that reported an alternation in the calcium metabolism and bone loss related to the length of time after menopause. Both the childhood and current milk consumption were also associated with a decreased risk: ORs were 0.4 (95% CI: 0.2-0.9) and 0.5 (95% CI: 0.3-1.0), respectively.

The laser speckle method is a new form of flowmetry that can obtain a two-dimensional distribution of blood flow in tissue. This method is a noncontact, simple, and rapid technique that may aid in the diagnosis of osteonecrosis. We investigated whether the subchondral bone blood flow within the femoral condyles of rabbits could be measured by the laser speckle method. The hydrogen washout method was chosen as a comparison technique because of its ability to allow repetitive measurements of blood flow in various conditions in one rabbit and because of its reliability, which already has been established. We simultaneously measured the bone blood flow in 20 femoral condyles of 10 rabbits with the laser speckle and hydrogen washout methods and found a significant correlation between the blood flow levels with use of these two methods. For the clinical application of the laser speckle method, we also investigated the influence of cartilage thickness on the measurements and the depth in the bone to which blood flow could be measured with this method. A cartilage thickness of 0.2 mm did not influence the measurement of the bone blood flow, and the depth in the bone to which the laser speckle method could be used was approximately 2 mm.

Human macrophage inflammatory protein-1 alpha (hMIP-1 alpha), also known as LD78, is a member of the chemokine/ intercrine family and an inhibitor of the proliferation of the hematopoietic stem cells in vitro. Using a specific monoclonal antibody, we observed significant localization of hMIP-1 alpha in eosinophilic myelocytes in human bone marrow. We further examined the expression of hMIP-1 alpha mRNA in human bone tissue by in situ hybridization. A high level of hMIP-1 alpha mRNA expression was detected in eosinophilic myelocytes in bone marrow, confirming these cells as the site of hMIP-1 alpha synthesis. hMIP-1 alpha mRNA expression was also detected in osteoblasts in the bone-remodeling sites, and osteoclasts were frequently observed in the vicinity of these osteoblasts. hMIP-1 alpha was also able to induce osteoclastogenesis on calcified matrices in the absence of any other osteotropic hormones. These results strongly suggest that hMIP-1 alpha is involved not only in the regulation of hematopoiesis but also in the modulation of bone remodeling.

We describe a quantitative assay for assessing tumor angiogenesis in vivo using basement membrane extracts (Matrigel). Nude mice were injected s.c. with liquid Matrigel mixed with HT1080 human fibrosarcoma cells. Since Matrigel rapidly forms a solid gel at body temperature, the gel containing tumor cells can be removed immediately and then processed for histological studies. Tumor angiogenesis was monitored quantitatively by measuring both the number and the total area of neovessels present in the gels using an image analyzer, which could be achieved approximately 72 hr later. Furthermore, HT1080 cell-conditioned medium, which may contain various tumor-derived factors, promoted the basement membrane degradation, migration, proliferation and tube formation of endothelial cells in vitro, as did Matrigel, although to a lesser extent. In addition, Northern blot analysis demonstrated that the amount of vascular endothelial growth factor (VEGF) mRNA in HT1080 cells was much higher than that in human fibroblasts or NIH3T3 cells. Our results suggest that angiogenesis observed in our assay may be due to the synergic effects of tumor angiogenic factors such as VEGF, and Matrigel. The advantages of our assay are: 1) it is possible to assess early angiogenesis quantitatively; and 2) this assay may be applicable for screening anti-angiogenic therapeutic agents to be used against human neoplasms.

Skeletal immature rabbits were used to study the pathogenesis of Osteochondritis Dissecans (OCD). Both histological studies and a radiographical examination were utilized after sagittal and coronal surgical chondral fractures were made in the femoral condyles cartilage. Serial microangiographies were performed in rabbits between 0 and 84 days after the chondral fractures were made. Analyses of the histology and microradiography findings suggest in either a coronal or sagittal direction, that avascular lesions like an experimental OCD occur as a sequence of chondral injury. A fracture in a wide pedicle of a stable cartilaginous flap with abundant cartilage canals heals in the usual way. However, a fracture in an unstable fragment with a small isthmus devoid of cartilage canals and of nutritious vessels, probably doesn't heal completely and a fragment closely resembling OCD is instead formed. An experimental OCD depends on the slender hinge of the flap and on the lack of stability in a rabbit's non-ossified epiphyseal cartilage. The damage to the cartilage canals and the rupture of vessels in the canals by a chondral fracture and the disturbance in the revascularization in the healing process by abnormal mechanical forces are thus most likely considered to be the main factor for OCD production.

This study performed to establish the most suitable method in one-day protocol and to evaluate clinical usefulness of 99mTc-tetrofosmin myocardial scintigraphy after dipyridamole infusion. Image quality and liver overlapping of the myocardial SPECT were evaluated in 107 patients with old myocardial infarction (42 cases), angina pectoris (53 cases) and others (12 cases). Left ventricular wall motion and coronary artery stenosis were compared to myocardial uptake score in 55 patients who received cardiac catheterization. The suitable image quality was acquired in early SPECT images using 259 MBq of 99mTc-tetrofosmin. The overlapping between inferior wall and liver uptake was able to minimize over 45 minutes interval from injection of 99mTc-tetrofosmin to data acquisition. The segments of normal wall motion had no perfusion defect of the myocardial SPECT in all cases (100% (148/148)). The segments of abnormal wall motion had decreased myocardial uptake of the myocardial SPECT (77% (24/31)). The agreement between coronary artery stenosis and decreased myocardial uptake was 96% (24/25) in right coronary artery, 87% (26/30) in left anterior descending coronary artery and 83% (19/23) in left circumflex coronary artery. These data suggests that image quality of dipyridamole loading 99mTc-tetrofosmin myocardial scintigraphy. Myocardial perfusion in the 99mTc-tetrofosmin myocardial scintigraphy is good correlation to both left ventricular wall motion and coronary artery stenosis.

Tissue inhibitors of metalloproteinases (TIMPs) play an important role in regulating the activity of matrix metalloproteinases (MMPs). Tumor cell invasion and metastasis closely correlate with the activities of two members of MMPs, MMP2 and MMP9, both of which degrade type IV collagen in basement membranes. We herein report that the treatment of HT1080 cells with 8-bromo-cAMP and other chemicals that activate cyclic adenylase activity induces the expression of TIMP1 and TIMP2 both at the mRNA and the protein levels and that this induction of TIMPs correlates with suppression of invasive phenotypes of HT1080 cells. Treatment with various cAMP-elevating reagents induced the expression of TIMPs and MMP2 in HT1080 cells, whereas the expression of MMP9 was not significantly affected. The protein amounts of TIMP1, TIMP2, and MMP2 secreted into the medium from HT1080 cells treated with 1 mM 8-bromo-cAMP were 7.9-, 9.3-, and 8.5-fold higher than those secreted from untreated cells, respectively. Induction of these mRNAs by 8-bromo-cAMP was blocked by HA1004, a protein kinase A inhibitor, but not by calphostin C, a protein kinase C inhibitor. Cycloheximide abolished the induction of TIMPs and MMP2 mRNAs by 8-bromo-cAMP, indicating that the induction depends on a newly synthesized protein(s) whose expression may be regulated by cAMP. Type IV collagenolytic activity and the invasiveness of HT1080 cells, both of which were suppressed by 8-bromo-cAMP, were efficiently restored when the cells were exposed to anti-TIMP antibodies, demonstrating the importance of the increased levels of TIMP1 and TIMP2 proteins for the cAMP-mediated suppression of both type IV collagenolytic activity and the invasiveness of HT1080 cells.

The histogenetic and biologic features of peripheral neuroepithelioma (PN) are still a matter of controversy. More detailed analyses might be facilitated by permanent PN cell lines. However, there have been only a few reports on the establishment of a PN cell line.We established and characterized a PN cell line and further carried out experiments for potentiality of drug-induced neural differentiation.The cultured cells were spindle shaped with slender cytoplasmic processes. The histologic features of transplanted tumors in nude mice were essentially the same as those of the original tumor. Immunohistochemically, the neuroectodermal characteristics of the cell line were demonstrated by positive stain for neuron specific enolase, glial fibrillary acidic protein, S-100 protein, neurofilament, and beta 2-microglobulin. Ultrastructurally, microtubules and dense core granules were observed in the cultured cells. Northern blot analyses revealed that c-myc was overexpressed in the cell line, whereas N-myc was not. The cell line showed neural differentiation after treatment with cAMP elevating reagents and nerve growth factor (NGF). The synergistic effects of cAMP analog and NGF on the neurite outgrowth were also observed. Furthermore, a cAMP-dependent protein kinase (PKA) inhibitor strongly blocked the action of cAMP analog, although it was not able to inhibit the same action of NGF. These findings suggested that the neurite outgrowth induced by the cAMP analog requires activation of PKA, whereas the same actions of NGF do not mediate such a pathway involving PKA activation.This is, to our knowledge, the first NGF responsive cell line derived from PN. This cell line might be valuable for further investigations of signal transduction pathways induced by cAMP analogs and NGF, for more precise analyses of the biological characteristics of PN, and finally, for the identification of differences between PN and several related tumors.

Myocardial relaxation at the diastolic phase was not evaluated by multi-gated myocardial SPECT, although myocardial contraction at the systolic phase was studied by percent wall thickening and Bull's eye methods. We make out a myocardial volume curve and report to evaluate the myocardial relaxation using multi-gated myocardial SPECT. The study population consisted of 3 normal human subjects (3 male, 32-37 years old), 10 idiopathic cardiomyopathy, 10 coronary artery disease and 1 hypertensive heart disease combined with aortic regurgitation. All cases were injected 555 MBq of 99mTc-tetrofosmin (Amersham Healthcare Corporation) intravenously at rest. A triple detector gamma-camera (GCA-9300A, Toshiba Medical, Japan) and a data processing computer (GMS-5500A, Toshiba Medical, Japan) were used in this study. A cardiac cycle (R-R interval) was divided by 16 frames (50-80 msec per 1 frame). Eight myocardial volume curves were calculated at the anterior wall, apex and inferior wall of the vertical long axis view and were calculated at the septal wall, apex and lateral wall of the horizontal long axis view, respectively. The patterns of the myocardial volume curves were classified into 5 patterns (Normal pattern (N), Delayed Contraction pattern (DC), Delayed Relaxation pattern (DR), Mixed pattern (M) and Normal pattern with Decreased amplitude (ND)). Myocardial uptake was evaluated visually of grading into severe hypertrophy (5), hypertrophy (4), normal (3), mild hypoperfusion (2), hypoperfusion (1) and perfusion defect (0). We compared patterns of the myocardial volume curves to myocardial uptake in the same segments. It was possible to detect myocardial edge of the total 16 frames with 50-60% threshold in the normal volunteer and in patients with hypertrophic cardiomyopathy and to make a myocardial volume curve. The region of the severe myocardial perfusion defect could be detected with 20% threshold in patients with old myocardial infarction. In comparison with myocardial volume curves and myocardial uptake, 74.6% in the N pattern had a normal uptake (3), 66.7% in the ND pattern had a normal uptake (3), 61.5% in the DC pattern had a hypoperfusion segment (0, 1 or fill-in to normal uptake), 44.4% in the DR pattern had a hypertrophic segment (4, 5 or fill-in to increased uptake). The pattern of myocardial volume curve indicates myocardial contractility and relaxation in each myocardial segment.

A case of giant cell reparative granuloma in a 16-year-old girl has been reported. This is the first reported case of giant cell reparative granuloma associated with enchondromatosis. In addition to the clinical features, the pathologic findings and differential diagnosis were also discussed.

In order to evaluate the effect of recombinant human erythropoietin (rHuEPO) on autologous blood transfusion in patients with rheumatoid arthritis (RA), we performed a phase II clinical trial in 65 RA patients undergoing elective surgery. rHuEPO was administered subcutaneously once a week and after observing erythropoiesis, autologous blood was collected. Fifty-seven of the 58 patients who completed treatment responded to rHuEPO and could donate more than 400 ml of autologous blood. Among them, 23 out of 28 patients undergoing total hip arthroplasty, 27 out of 28 undergoing total knee arthroplasty and 1 out of 1 undergoing spinal surgery did not need homologous blood transfusion perioperatively. During rHuEPO treatment, no significant changes of clinical parameters of RA activity were observed. Two patients discontinued the treatment because of mild and transient side effects. These results indicate that subcutaneous rHuEPO is safe and effective in eliminate the need for homologous blood transfusion, even in anemic RA patients undergoing elective orthopedic surgery.

The newly developed 99mTc-labeled renal imaging agent, mercaptoacetyltriglycine (MAG3), is a tubular secreted compound. It is expected the most suitable agent to measure renal function and image urinary tract because of the excellent imaging properties and availability of 99mTc and preferred biological qualities of radiohippurates. We examined 15 patients (8 males, 7 females, mean age 55 years old) with renal and urinary tract disease. After drinking 400 ml of water, 185, 370 or 555 MBq of 99mTc-MAG3 was injected with the supine position. Dynamic scans were acquired for 20 minutes on a large field of view gamma camera (ZLC-7500 Siemens Medical Inc.) linked to a computer system (Scintipac-2400 Shimadzu Co.) in a 64 x 64 matrix. Regions of interest (ROI) were placed around each kidney. Curves of 12 seconds frame rate were generated from all two ROIs over the 20 minutes study. The time to peak count (Tmax) and time from peak count to a half count (T1/2) were calculated from the curve data. 99mTc-DTPA renal imaging was examined to the same patients with the same method like 99mTc-MAG3. The renogram of 131I-orthoiodohippurate (OIH) was performed with probe method. The excellent quality of the renal image was obtained with 185-555 MBq of 99mTc-MAG3 compared with the same dose of 99mTc-DTPA respectively. The diagnostic value of 99mTc-MAG3 renal imaging was as well as 99mTc-DMSA renal imaging. The Tmax of 99mTc-MAG3 renogram was same value of that of both 99mTc-DTPA and 131I-OIH renogram. The T1/2 of 99mTc-MAG3 renogram was significantly lower than that of 99mTc-DTPA renogram. The T1/2 of 99mTc-MAG3 renogram was slightly higher that of 131I-OIH renogram. We conclude that the renal imaging of 99mTc-MAG3 was able to acquire the most excellent image and the proper renogram pattern as same as 131I-OIH renogram.

Preoperative blood reservation for autologous blood transfusion generally causes anemia. We performed a double blind controlled study to determine the optimal dose of subcutaneous rHuEPO (recombinant human erythropoietin, KRN5702) for preventing preoperative anemia due to blood reservation. Patients received KRN5702 subcutaneously once a week in a doses of 12000, 24000 or 36000 IU by a double blind technique. After storage of 1200 ml of their own blood right before surgery, their hemoglobin (Hb) averaged about 1 and was about 0.5 g/dl lower than the level before administration of KRN5702 in doses of 12000 and 24000 IU, respectively. This fall was significant. In patients receiving KRN5702 in a dose of 36000 IU, the level of Hb rose instead of a fall; Hb immediately before surgery was 1.1% higher than that before administration which, however, was not significant. This elevation indicates a possibility of abnormal elevation of Hb at this dose. The mean Hb value right before surgery was significantly lower in patients receiving 12000 IU KRN5702 than in patients of the other two groups. The recovery rate of Hb was an indicator to reflect improvement of anemic conditions, and increased as the dose increased after the blood reservation. The rate in the 12000 IU group was significantly lower than that in the other two groups; there was not much difference between the other two rates. We estimated that to reserve 1200 ml of autologous blood, 24000 IU of KRN5702 is adequate but not excessive. One patient receiving 24000 IU showed side effects including an elevation of body temperature, rash, and edema.

A Multi-gated SPECT was acquired commonly in 64 x 64 matrix and 8 frames per cardiac cycle (64 x 64/8F). But it was not established that 64 x 64 matrix and 8 frames per cardiac cycle were the most suitable in multi-gated SPECT. Five normal volunteers were examined multi-gated 201Tl SPECT with 5 acquisition modes of 128 x 128 matrix/16 frames, 128 x 128/8F, 64 x 64/32F, 64 x 64/8F using multi-detector SPECT system (GCA-9300). And we calculated percent wall thickening (%WT) [%WT = (ES counts - ED counts)/ED counts] in 9 cases with 64 x 64/8F. The images quality of both 128 x 128/16F and 128 x 128/8F was not clear in compared with images of both 64 x 64/16F and 64 x 64/8F, because the end-diastolic phase of 128 x 128/16F images showed a decreased uptake of 201Tl in the antero-apical region. Although 64 x 64 (8) images had only 8 frames per cardiac cycle, we could observe systolic and diastolic phase and we could calculate %WT. The %WT (M +/- SD) of horizontal long axis images were 48 +/- 15 (sept. basal), 48 +/- 19 (sept. apical), 65 +/- 29 (apex), 49 +/- 22 (lat. apical) and 40 +/- 15 (lat. basal).(ABSTRACT TRUNCATED AT 250 WORDS)

In the electrodiagnosis of cubital tunnel syndrome, the abductor digiti minimi (ADM) muscle is frequently used to evoke compound muscle action potentials (CMAP) in order to measure the motor nerve conduction velocity (MNCV) in the ulnar nerve. Slowing of the MNCV across the elbow is a major criteria in the electrodiagnosis of cubital tunnel syndrome. However, in advanced cases, slowing of the MNCV is also seen in the forearm segment, thus, localization of the nerve palsy is sometimes difficult. We evoked CMAP from the flexor carpi ulnaris (FCU) muscle to assess the value of electrophysiological examination of this muscle in the diagnosis of cubital tunnel syndrome. CMAP from the FCU muscle could be evoked in many cases when CMAP from the ADM muscle could not. Also, the motor terminal latency (MTL) of the FCU was prolonged in proportion to the slowing of the MNCV across the elbow. Therefore, electrophysiological examination of the FCU muscle is useful in the diagnosis of cubital tunnel syndrome, especially when intrinsic muscle atrophy is severe and the CMAP from these muscles cannot be evoked.

Although we could acquire a detailed 512 x 2048 matrix whole body scintigraphy, the 512 x 2048 matrix whole body scintigraphy was divided to the upper and lower half of the body, because a many of CRT system displayed only 1024 x 1024 matrix with non interlace mode. We made 12 dots of normal vertical image distance to 0 dot with laser imaging system (Li-10 Konica medical inc.), and we printed these divided whole body images in the four partition of the film. The lead bar phantom (interval from 6 mm to 3 mm) filled with 99mTcO4- was studied by both 512 x 2048 matrix whole body scanning mode and 256 x 1024 whole body scanning mode in the basic study. And the distance between the lead bar phantom and the gamma camera was changed from 10 mm to 100 mm. We studied 41 patients with metastatic bone tumor (14 breast cancer, 7 lung cancer, 7 prostate cancer, 5 others, 6 unknown origin) clinically. However the 512 x 2048 matrix whole body scan was better quality of images than 256 x 1024 matrix whole body scan at 100 mm distance in the basic study. The abnormal uptake of metastatic sites was shown equally in both 512 x 2048 and 256 x 1024 matrix whole body scintigraphy. The 512 x 2048 matrix whole body scan was better quality of images than 256 x 1024 matrix whole body scan in 26 out of 41 patients, equal in 10 out of 41 patients and worse in 3 out of 41 patients. The matrix size of 512 x 2048 matrix whole body scintigraphy (0.98 mm2) was smaller than that of 256 x 1024 matrix whole body scintigraphy (1.95 mm2).(ABSTRACT TRUNCATED AT 250 WORDS)

99mTc-tetrofosmin is a lipophilic, cationic diphosphine which has been developed for myocardial imaging. We examined 9 patients with ischemic heart disease including 3 angina pectoris (AP), 4 old myocardial infarction (OMI), 1 AP with OMI and 1 syndrome X. One patient was examined before and after operation. 370 MBq of 99mTc-tetrofosmin was injected during exercise and 740 MBq at rest. And 74 MBq of 201Tl myocardial exercise and redistribution scintigraphy was also performed to compare with 99mTc-tetrofosmin myocardial scintigraphy. SPECT, multiple gated SPECT and anterior planar images were obtained in all cases. We calculated percent wall thickening (%WT) using multiple gated SPECT images. There was a decreased lung uptake in 99mTc-tetrofosmin planar images compared to 201Tl myocardial scintigraphy. Liver and Biliary system uptake in 99mTc-tetrofosmin images was decreased with intake of milk. Segmental comparison of SPECT images showed an agreement in 9/10 of the segment between 201Tl and 99mTc-tetrofosmin. We could obtain excellent quality of multiple gated SPECT images in all patients. We could calculate percent wall thickening (%WT) in all patients. We conclude that 99mTc-tetrofosmin myocardial scintigraphy should provide usefulness for detection of ischemic myocardium as same as 201Tl myocardial scintigraphy, although the biologic characteristics of two agents were different. These data and excellent quality of multiple gated SPECT images suggest that 99mTc-tetrofosmin is a new 99mTc agent for evaluation of patients with ischemic heart disease.

This is the first report of the process of formulation of a cervical osteophyte causing dysphagia. The patient had ankylosing spinal hyperostosis and OPLL and was followed radiographically for a long time before the onset of dysphagia. The radiological observation suggested that dysphagia was produced when the immobile part of the esophagus was compressed by the anterior projecting cervical osteophyte. The immobility of the esophagus is an important factor in determining whether dysphagia occurs. Another possible contributing factor to dysphagia in this patient was the ossification of the cervical anterior and posterior longitudinal ligaments. The OPLL affected intervertebral segmental motion and induced the formation of anterior projecting cervical osteophytes.

Number of victims of nontraumatic avascular osteonecrosis of the femoral head (ONFH) is believed to have increased world widely.this is partly owing to an improved awareness of the condition and developtment of diagnosis imaging techniques sensetives to the condition.

We measured evoked mixed nerve action potentials (EMNAP) in the forearm segment of 108 hands as an indicator of retrograde degeneration in the carpal tunnel syndrome (CTS). This was made by recording at the elbow and stimulating the median nerve 4 cm proximal to the wrist crease. The amplitude of this EMNAP decreased in proportion to the prolongation of the motor terminal latency (MTL) or the decrease in the amplitude of the compound muscle action potential (CMAP) from the abductor pollicis brevis (APB) muscle. This suggests that retrograde degeneration progresses as the severity of CTS increases. This decrease was also in proportion to the decrease in the motor nerve conduction velocity (MNCV) in the forearm. Both MNCV in the forearm and EMNAP can indicate retrograde degeneration in CTS, but the MNCV cannot be measured when muscle atrophy is severe and the CMAP cannot be elicited. Therefore, we believe that EMNAP is a more useful measure of the retrograde changes in CTS.

First of all, I would like to mention the principle of the treatment of the osteonecrosis of the femoral head based on the following clinical observations.

We have operated on 42 patients with bone metastasis from various primary tumors over the past fourteen years. The number of operated cases of metastasis secondary to thyroid and breast carcinoma was especially high, since patients with such types of cancer have prolonged life expectancies in general. The sites of these operation were as follows: 27 in the spine, 12 in the femur, 2 in the tibia, and 1 in the humerus.

We have used an extract of basement membranes which can be reconstituted into a biologically active gel matrix composed predominantly of collagen IV, laminin, nidogen, and heparin sulfate proteoglycan, in order to study the mechanisms involved in tumor cell invasion. When layered onto a porous filter in a Boyden chamber, the gel forms a barrier to the passage of normal cells. Malignant cells are able to cross this layer when the conditioned medium of NIH 3T3 cells is used as a chemoattractant to stimulate cell migration. A variety of human tumor cells have thus been studied in this system and we find a high correlation between their invasiveness in vitro and their malignant behavior as exhibited in vivo. We have used this in vitro invasion assay to test for factors which might inhibit tumor cell invasion. Collagenase IV is produced by malignant cells and is thought to be required for invasion. Indeed, inhibitors of this enzyme have demonstrated reduced tumor cell invasiveness. One site of five amino acids, on the B1 chain, which has been shown to promote cell adhesion, migration and binding to laminin receptor, was found to inhibit the invasion of tumor cells. In addition, factors which elevated cAMP levels were also able to suppress the invasiveness of tumor cells. These data suggest that the assay system described herein can be successfully utilized to study the invasive activity of tumor cells and those factors that may inhibit the spread of malignant cells.

Tobramycin sulfate powder (1.2 g) was mixed with Palacos polymethylmethacrylate (PMMA) bone cement (40 g) to produce 100 discs containing 5.9 mg tobramycin per disc. These discs were used to evaluate the inhibition of bacterial adhesion to an antibiotic-laden biomaterial. Tobramycin-impregnated PMMA discs and control discs containing no tobramycin were exposed in vitro to Staphylococcus epidermidis. Colonization was quantitated using plate count techniques and electron microscopy. Tobramycin-impregnated surfaces reduced adhesive bacteria colonization by 1 log relative to control discs. These observations suggest that tobramycin-impregnated PMMA may not be significantly effective in preventing colonization of the biomaterial substratum and PMMA may be a poor choice as a drug delivery vehicle in biomaterial and compromised tissue-centered infections.

The role of the cartilage canals of the epiphysis was investigated by studying the process of initial enchondral osteogenesis of the secondary ossification center using decapitated femoral heads of one day old Japanese rabbits. Specimens of decapitated femoral heads returned to the original acetabulum demonstrated granulation tissue invasion at ten days after birth, followed by the normal process of enchondral osteogenesis of the secondary ossification centers. In contrast, specimens of heads inserted in the peritoneal cavity to prevent invasion of granulation tissue showed no enchondral osteogenesis of the secondary ossification center, and the major portion of the decapitated femoral head was replaced by necrotic chondrocytes and empty lacunae. The results of this study confirm that the cartilage canal is important not only in the nutrition of the chondroepiphysis but also in the initial enchondral osteogenesis of the secondary ossification center.

Using a whole-limb graft model in rats, morphologic changes and variations in the expression of Ia antigen on epidermal cells were investigated in the allografted skin during acute rejection. BN right limbs were transplanted to F344 recipients. Skin tissues were excised during acute rejection on days 1, 3, 5, 7, and 9 after the transplantation. Sections were examined for Ia antigen expression using immunohistologic techniques, and in situ quantification of Ia antigen was made using an immunogold method. Epidermal keratinocytes expressed Ia antigen before the grafts were rejected and the amount of Ia antigen expression increased and exceeded the amount of Ia antigen of Langerhans cells during the course of rejection. The progressive increase in class II antigen expression on EKs correlated with the appearance and relative accumulation of dermal lymphocytic cells. On the other hand, Ia antigen was not expressed on vascular endothelial cells during rejection. Our results suggest that the Ia-positive keratinocytes can serve as target cells in skin rejection of limb allografts. The immunogold technique we used seems most pertinent for a quantitative examination of cell-surface antigens in situ.

The authors studied how bone reacts to hydroxyapatite (HAP) and alumina (Al 2 O 3 ) in osteoporosis experimentally induced in a rat animal model by resection of the sciatic nerve. Sixteen and 32 weeks after resection of the sciatic nerve, implants of HAP and Al 2 O 3 were inserted into rat tibias. The reaction of bone to HAP was compared with its reaction to Al 2 O 3 , which is bioinert, as a control. The results were analyzed using the affinity index (the length of bone directly opposed to the implant without an intervening fibrous membrane/total length of the bone-implant interface × 100%). In the rats left for 16 weeks after resection of the sciatic nerve the index values 24 weeks after insertion of HAP and Al 2 O 3 were 96.9 ± 2.4% (N = 8) and 75.2 ± 22.2% (N = 8), respectively. The index value of HAP was 97.2 ± 2.6% (N = 12) and of Al 2 O 3 was 86.9 ± 15.4% (N = 12) at 24 weeks after insertion, when the rats were first left for 32 weeks, after sciatic nerve resection. Bone was demonstrated to have a superior affinity to HAP, compared with Al 2 O 3 ( P