Effects of short-term fasting on the Akt-mediated pathway involved in protein metabolism in chicken skeletal muscle.

In the present study, we show that short-term (4 h) fasting significantly decreased the levels of protein synthesis-related factors such as the plasma insulin concentration, skeletal muscle pAkt, and pS6 levels in 2-wk-old chickens (P < 0.05). An intravenous injection of insulin significantly elevated the contents of pAkt and p-S6 in the skeletal muscle (P < 0.01). These findings suggest that decreasing the plasma insulin causes the downregulation of the Akt/S6 pathway in chicken skeletal muscle under short-term fasting conditions. However, protein synthesis was not significantly affected by short-term fasting. In addition, no significant change was observed in the levels of proteolysis-related factors such as plasma Nτ-methylhistidine, phosphorylated forkhead box class O, and muscle ring finger-1 during 4-h fasting, indicating that short-term fasting does not induce skeletal muscle proteolysis in chickens. Interestingly, atrogin-1 expression significantly increased after 2-h fasting (P < 0.05), and insulin injection significantly reversed the fasting-induced atrogin-1 expression in chicken skeletal muscle (P < 0.01). Collectively, these findings suggest that short-term fasting downregulates the insulin-stimulated Akt/S6 pathway but does not significantly affect protein synthesis and proteolysis in chicken skeletal muscle, and that atrogin-1 expression is upregulated in a FOXO1-independent manners.

IL-28B (IFN-λ3) and IFN-α synergistically inhibit HCV replication.

Genetic variation in the IL-28B (interleukin-28B; interferon lambda 3) region has been associated with sustained virological response (SVR) rates in patients with chronic hepatitis C treated with peginterferon-α and ribavirin. However, the mechanisms by which polymorphisms in the IL-28B gene region affect host antiviral responses are not well understood. Using the HCV 1b and 2a replicon system, we compared the effects of IFN-λs and IFN-α on HCV RNA replication. The anti-HCV effect of IFN-λ3 and IFN-α in combination was also assessed. Changes in gene expression induced by IFN-λ3 and IFN-α were compared using cDNA microarray analysis. IFN-λs at concentrations of 1 ng/mL or more exhibited concentration- and time-dependent HCV inhibition. In combination, IFN-λ3 and IFN-α had a synergistic anti-HCV effect; however, no synergistic enhancement was observed for interferon-stimulated response element (ISRE) activity or upregulation of interferon-stimulated genes (ISGs). With respect to the time course of ISG upregulation, the peak of IFN-λ3-induced gene expression occurred later and lasted longer than that induced by IFN-α. In addition, although the genes upregulated by IFN-α and IFN-λ3 were similar to microarray analysis, interferon-stimulated gene expression appeared early and was prolonged by combined administration of these two IFNs. In conclusion, IFN-α and IFN-λ3 in combination showed synergistic anti-HCV activity in vitro. Differences in time-dependent upregulation of these genes might contribute to the synergistic antiviral activity.

Associations of vertebral deformities and osteoarthritis with back pain among Japanese women: the Hizen-Oshima study.

UNLABELLED: We examined the spinal distribution of the types of vertebral deformities and the associations of vertebral deformities and osteoarthritis with back pain in Japanese women. Midthoracic and upper lumbar vertebrae were more susceptible to deformity. Vertebral deformity and osteoarthritis were frequent and were associated with back pain. INTRODUCTION: Vertebral fractures due to osteoporosis and osteoarthritis are both common and significant health problems in aged people. However, little is known about the descriptive epidemiology of the individual deformity types and the relative clinical impact in women in Japan. METHODS: Lateral radiographs were obtained from 584 Japanese women ages 40 to 89 years old. Deformities were defined as vertebral heights of more than 3 standard deviations (SDs) below the normal mean. Osteoarthritis was defined as Kellgren-Lawrence (KL) grade 2 or higher. Information on upper or low back pain during the previous month was collected by questionnaire. We compared the spinal distribution of the three types of vertebral deformities (wedge, endplate, and crush) typical of fractures and examined the associations of number and type of vertebral deformities and osteoarthritis with back pain. RESULTS: Fifteen percent of women had at least one vertebral deformity and 74% had vertebral osteoarthritis. The prevalence of upper or low back pain was 30.1%. Deformities were most common in the midthoracic and upper lumbar regions and wedge was the frequent type, followed by endplate and crush. Multiple logistic regression analysis showed that the odds of back pain was 3.0 (95% CI 1.5-6.3) times higher for women with a single wedge deformity and 3.2 (95% CI 1.0--0.6) times higher for women with two or more wedge deformities, compared to women with no wedge deformity. Vertebral osteoarthritis was associated with back pain (OR 1.8, 95% CI 1.1-2.9), independent of other covariates including age and deformities. CONCLUSION: Our results in this group of Japanese women are similar to and consistent with results reported previously in other populations of Japanese and Caucasians.

Osteoporotic changes of subchondral trabecular bone in osteoarthritis of the knee: a 3-T MRI study.

SUMMARY: Subchondral trabecular bone structure was analyzed in knee osteoarthritis (OA) patients using 3-T MRI to investigate structural features of subchondral trabecular bone of knee OA. With OA progression, osteoporotic changes were observed in the lateral joint, showing a higher correlation than sclerotic changes in the medial joint. INTRODUCTION: To investigate structural features of subchondral trabecular bone of knee osteoarthritis (OA). METHODS: Sixty knees with KL grade 0-4 (all female) were examined. Fast imaging employing steady-state acquisition-cycled phases (FIESTA-c) and FatSat Spoiled gradient recalled acquisition in the steady state (SPGR) images were acquired by 3-T MRI. At four sites (the medial femur, medial tibia, lateral femur, and lateral tibia), subchondral trabecular bone structure was analyzed by FIESTA-c imaging, cartilage area was measured by SPGR imaging, and their correlation was analyzed. In addition, the subjects were classified into four groups from the cartilage area measured by SPGR imaging, and subchondral trabecular bone structure in each group was compared. RESULTS: As cartilage area decreased in the medial joint, bone volume fraction and trabecular thickness in the medial tibia increased, and bone volume fraction, trabecular thickness, number, and connectivity in the lateral femur and lateral tibia decreased (r ≥ 0.4 or ≤-0.4, p ≤ 0.001). Compared to medially, the changes laterally showed a higher correlation. When the medial-lateral ratio of trabecular thickness in the tibia was determined, it had the highest correlation coefficient (r=-0.7, p < 0.001). These changes were not significantly detected in the early stage. CONCLUSIONS: To more sensitively detect OA changes in subchondral trabecular bone structure, a focus on osteoporotic changes in the lateral joint and the medial-lateral ratio would be useful. Detectability of early OA remains unknown, but based on a strong correlation with the degree of OA progression, trabecular structural analysis of subchondral bone may be a useful parameter to evaluate OA severity and evaluate treatment.

Beneficial effect of risedronate for preventing recurrent hip fracture in the elderly Japanese women.

SUMMARY: A 36-month observational study compared the incidence of unaffected side hip fracture in Japanese female osteoporosis patients with a history of hip fracture between 173 patients receiving risedronate and 356 risedronate-untreated controls. New hip fractures were significantly less frequent in the risedronate group, suggesting a preventive effect in high-risk patients. INTRODUCTION: The purpose of this study was to investigate the preventive effect of risedronate on second hip fracture immediately following a first hip fracture in Japanese female osteoporosis patients with unilateral hip fracture. METHODS: We conducted a prospective matched cohort study in 184 patients treated with risedronate and 445 patients not receiving risedronate after discharge from hospital. Both groups were followed-up for 36 months, and the incidence of unaffected side hip fracture and the frequency of adverse events were assessed. RESULTS: Efficacy could be investigated in 173 patients from the risedronate group and 356 patients from the control group. Hip fracture was detected in 5 and 32 patients, respectively. Kaplan-Meier estimates of the 36-month fracture incidence were 4.3% in the risedronate group and 13.1% in the control group (P = 0.010, log-rank test). The hazard ratios (95% confidence intervals) obtained by univariate and multivariate analysis were 0.310 (0.121-0.796) and 0.218 (0.074-0.639), respectively, indicating a significantly lower incidence of unaffected side hip fracture in the risedronate group. Adverse events occurred in 38 patients (48 events) from the risedronate group and 94 patients (108 events) from the control group, with serious adverse events in 21 patients (26 events) and 78 patients (88 events), respectively. CONCLUSIONS: No significant differences were observed between the two groups. The incidence of unaffected side hip fracture was significantly lower in the risedronate group. Accordingly, risedronate may have a preventive effect on hip fracture in high-risk Japanese female osteoporosis patients for fracture with a history of unilateral hip fracture.

In vivo structural analysis of subchondral trabecular bone in osteoarthritis of the hip using multi-detector row CT.

OBJECTIVE: With developments in clinical computed tomography (CT), in vivo analysis of patients' bone microstructure has become increasingly possible. We analyzed the subchondral trabecular bone of hip osteoarthritis (OA) patients using multi-detector row CT (MDCT) to closely examine the structural changes that occur as OA progresses. DESIGN: 47 female hip joints were studied: 20 with OA secondary to hip dysplasia (11 advanced OA, nine early-moderate OA), seven with hip dysplasia without OA, and 20 normal. The images' maximal spatial resolution was 280 × 280 × 500 µm. Regions of interest (ROIs) were the subchondral trabecular bones of the acetabulum and femoral head. Measurement parameters were bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular separation (Tb.Sp), structure model index (SMI), trabecular bone pattern factor (TBPf), Euler's number, and degree of anisotropy (DA). Relationships between joint space volume and these parameters were analyzed. RESULTS: With decreasing joint space, Tb.Th and BV/TV increased, and Tb.Sp, Tb.N, SMI, TBPf, and DA decreased significantly. The microstructures were significantly different between the early to advanced OA groups and the normal and dysplasia groups; there was no significant difference between the normal and dysplasia groups. CONCLUSIONS: Changes of subchondral trabecular bone structure in OA could be evaluated using MDCT, despite imperfect spatial resolution and limited accuracy. Trabecular bone thickening and associated structural changes may be closely related to OA. Changes were observed in early to advanced OA, but not in dysplasia. This method may help to further elucidate OA pathogenesis, determine the therapeutic strategy, and evaluate therapy.

Bone mineral densities in patients with developmental dysplasia of the hip.

UNLABELLED: Bone mineral density (BMD) of the lumbar spine, ultradistal radius, and calcaneus were significantly higher in the developmental dysplasia of the hip (DDH) patients than in the controls. Therefore, our data suggest that BMDs at different skeletal sites are greater in patients with DDH than in healthy women. INTRODUCTION: DDH has been acknowledged as a potentially preosteoarthritic condition that results in the development of hip osteoarthritis. Patients with DDH have been reported to have abnormal morphology of the pelvis and spine. Additional research, including that of bone quality, needs to be conducted to elucidate the pathogenetic mechanism of this disease. We therefore sought to determine whether BMD differs between healthy women and women with DDH. METHODS: We measured BMD in 40 women who were scheduled to undergo pelvic osteotomy for DDH (average age, 45.3 years) and in 31 healthy women used as age-matched controls (average age, 47.5 years). BMDs of the lumbar spine, radius, and calcaneus were measured. RESULTS: BMDs of the lumbar spine, ultradistal radius, and calcaneus were significantly higher in the DDH patients than in the controls. CONCLUSIONS: Therefore, our data suggest that BMDs at different skeletal sites are greater in patients with DDH than in healthy women.

Potentiation of glucocorticoid receptor (GR)-mediated signaling by the immunosuppressant tacrolimus in rheumatoid synoviocytes.

OBJECTIVE: The immunosuppressant tacrolimus is known to enhance many aspects of glucocorticoid. In this study, we investigated the effects of tacrolimus on glucocorticoid receptor (GR) signaling using rheumatoid fibroblast-like synoviocytes (RA-FLS). METHODS: The nuclear translocation of GR was analyzed by immunocytochemistry. The DNA binding activity of p65 was assayed by a functional ELISA kit using nuclear extracts. GR-associated FK506-binding protein-51 (FKBP-51) was analyzed by Western blotting following immunoprecipitation of glucocorticoid receptor (GR) complexes. RESULTS: High concentrations (10-7M) of Dexamethasone (Dex) induced GR translocation to the nucleus in RA-FLS. However, the nuclear GR translocation did not occur with low concentrations of Dex (10-9M). Tacrolimus treatment of RA-FLS results in potentiation of GR translocation to the nucleus even in the presence of a low concentration of Dex (10-9M). GR-associated FKBP-51 decreased after tacrolimus treatment. Furthermore, tacrolimus also decreased the IL-1Beta-induced DNA binding activity of p65, a subunit of NF-KappaB, in the presence of 10-9 M of Dex. CONCLUSION: These data suggest that tacrolimus exerts anti-inflammatory properties by potentiating the GR signaling through the GR-immunosuppressant-binding proteins (immunophilins) interaction and its nuclear transport in rheumatoid synovium.

Sweat antigen induces histamine release from basophils of patients with cholinergic urticaria associated with atopic diathesis.

BACKGROUND: We previously demonstrated that the semipurified human sweat antigen causes skin reactions and histamine release from basophils via specific IgE in patients with atopic dermatitis (AD). Patients with cholinergic urticaria (ChU) also develop skin reactions and histamine release of basophils in response to autologous sweat. OBJECTIVES: To study whether or not patients with ChU share sensitivity for the sweat antigen with patients with AD and to study the clinical characteristics among patients with ChU and the relationship with histamine-release activity of basophils. METHODS: The sweat antigen that induces histamine release from basophils of patients with AD was prepared by Con-A, anion-exchange and reverse-phase chromatography. Relationships between histamine-release activity against the sweat antigen and clinical features of patients with ChU were analysed. RESULTS: Twenty-three of 35 patients with ChU showed > 5% net histamine release in response to the semipurified sweat antigen, whereas none of healthy controls did so. In patients with ChU, histamine release in response to semipurified sweat antigen significantly correlated with the level of serum IgE and eosinophil numbers in peripheral blood. Incidence of each atopic disease in patients with ChU tended to be higher than in the general Japanese population. When the patients were categorized according to their responses in the histamine release test, the positive group tended to show a higher incidence of AD and bronchial asthma compared with the negative group. CONCLUSIONS: ChU and AD may share hypersensitivity to common antigens in sweat. The sweat allergy and atopic diathesis are associated with each other.

Rotational acetabular osteotomy with a resection of the lateral edge of the acetabulum for the treatment of advanced coxarthrosis.

INTRODUCTION: We compared the functional and radiological results of a rotational acetabular osteotomy (RAO) with and without a resection of the lateral edge of the acetabulum. The purpose of the resection was to obtain good joint congruency. MATERIALS AND METHODS: RAO was performed on 71 hips to treat advanced coxarthrosis caused by acetabular dysplasia. RAO without a resection (non-resection group) was performed in 54 patients (57 hips) with a median age of 43.1 years. The remaining 14 patients (14 hips), who had a median age of 44.6 years, received RAO with a resection of the lateral edge of the acetabulum (resection group). RESULTS: The average postoperative total hip joint score was better than the average preoperative score in the non-resection group (P < 0.001), but not in the resection group. In the resection group, all hips displayed progressive osteoarthritic change and ten hips had chondrolysis of the hip joint and a collapse of the transferred acetabulum within 3 years. In the non-resection group, 15 hips showed progressive osteoarthritic change, 24 hips had no change, and 18 hips showed a decrease in the osteoarthritic stage. CONCLUSION: Our findings demonstrated that resection of the lateral edge of the acetabulum is not a useful adjunct to the RAO procedure for the treatment of advanced coxarthrosis.

Rotational acetabular osteotomy for advanced osteoarthritis secondary to developmental dysplasia of the hip.

We evaluated the results of rotational acetabular osteotomy in 44 hips (42 patients) with advanced osteoarthritis secondary to developmental dysplasia. The mean age of the patients at surgery was 43.4 years (30 to 59) and the mean follow-up was 12.1 years (8 to 19). The mean Merle d'Aubigné clinical score improved from 10.8 points (8 to 15) pre-operatively to 13.5 points (6 to 18) at follow-up. Radiologically, this procedure produced adequate improvement regarding cover of the femoral head. At follow-up, the osteoarthritic stage assessed using the Japanese Orthopaedic Association grading, was improved in 11 hips (25%), unchanged in 22 (50%) and had progressed in 11 (25%). The mean pre-operative roundness index of the femoral head was significantly different in the 33 hips which had improved or maintained their osteoarthritic stage compared with the 11 which had progressed (53.7% vs 63.7%; p < 0.001). Osteoarthritis with a round femoral head is considered to be an indication for rotational acetabular osteotomy, even in advanced stages of the disease.

Barium aspiration showing crazy-paving appearance on high-resolution computed tomography.

We encountered a case showing a crazy-paving appearance on high-resolution computed tomography (HRCT), which mimicked various pulmonary diseases, e.g. pulmonary alveolar proteinosis, or non-specific interstitial pneumonia, but which we ultimately determined to be due to barium aspiration. The initial radiological findings were considered to be most likely due to pulmonary alveolar microlithiasis, because the crazy-paving appearance had high density, resembling calcification. However, distribution was limited to the area of dependent drainage of the right lung. Since a barium swallowing study had been performed 2 weeks before the CT examination, and since the follow-up HRCT showed improvement we reached a diagnosis of barium aspiration. Although barium aspiration may yield findings similar to other diseases with crazy-paving appearance, it is possible to identify it because of the limited distribution and high density.

Pharmacokinetic study of darbepoetin alfa: absorption, distribution, and excretion after a single intravenous and subcutaneous administration to rats.

KRN321 is a hyperglycosylated analogue of recombinant human erythropoietin (rHuEPO, epoetin alfa), and its absorption, distribution, and excretion have been studied after a single intravenous and subcutaneous administration of 125I-KRN321 at a dose of 0.5 microg kg-1 to male rats. The half-lives of immunoreactive radioactivity in the terminal phase after intravenous and subcutaneous administration were 14.05 and 14.36 h, respectively, and the bioavailability rate after subcutaneous administration was 47%. The total radioactivity in tissues was lower than that in the serum in all tissues excluding the thyroid gland and skin at the injection site (subcutaneous administration). The maximum concentrations were observed in the bone marrow or skin at the injection site followed by the thyroid gland, kidneys, adrenal glands, spleen, lungs, stomach and bladder. The radioactivity found in trichloroacetic acid-precipitated fractions suggested that a high-molecular weight compound, unchanged or mixed with endogenous protein, distributed to the tissues after administration. The whole-body autoradiographic findings in both groups were in agreement with the tissue distribution mentioned above. The blood cell uptake of KRN321 was low for both groups. The excretion ratios of radioactivity into urine and faeces up to 168 h were 71.4 and 14.1% after the intravenous administration and 74.9 and 12.0% after the subcutaneous administration. There was no difference in the excretion profile of radioactivity between the two groups.

Nanobacteria-like particles in human arthritic synovial fluids.

We investigated the existence of nanosize particles in synovial fluids of rheumatoid arthritis and osteoarthritis patients. These specimens were cultured under mammalian cell culture conditions (37 degrees C; 5% CO2/95% air) for a long period. After about 2 months, many nanoparticles appeared and they gradually increased in number and in size. The nanobacteria-like particles exist in synovial fluids of arthritis patients. The possibility of their existence and pathogenesis in various diseases should be verified cautiously.

Postoperative changes in protein-induced vitamin K absence or antagonist II levels after hepatectomy in patients with hepatocellular carcinoma: relationship to prognosis.

BACKGROUND: alpha-Fetoprotein (AFP) has been used as a marker for hepatocellular carcinoma (HCC). However, AFP levels are often high in patients with chronic hepatitis or cirrhosis. Protein-induced vitamin K absence or antagonist II (PIVKA-II) is more sensitive for the diagnosis of HCC and prediction of patient survival. Changes in these markers after treatment may reflect treatment curability and patient outcome. METHODS: We conducted a retrospective analysis of prognosis of 63 HCC patients with high preoperative levels of AFP and PIVKA-II who underwent hepatectomy and examined the relationship between postoperative changes in both markers at 1 month and patient survival. Subjects were divided into three groups according to changes in these tumour markers after hepatectomy: normalization (N) group, decreased but still above the normal level (D) group and unchanged (U) group. RESULTS: There were no significant differences in the numbers of patients who developed tumour recurrence between changes in AFP and PIVKA-II. Survival analysis showed no significant differences in tumour-free and overall survivals between groups with respect to AFP level. The PIVKA-II-N group showed significantly better tumour-free and overall survival compared with the D and U groups (p<0.01). Multivariate analysis that included other prognostic factors identified changes in PIVKA-II level as a significant and independent prognostic factor associated with overall survival. DISCUSSION: Although changes in AFP did not correlate with patient prognosis, normalization of PIVKA-II was significantly associated with good patient survival after hepatectomy. Normalization of PIVKA-II after hepatectomy reflected the efficacy of treatment and is a suitable predictor of prognosis in HCC patients.

Establishment of a clonal human mesenchymal cell line that retains multilineage differentiation capacity from a spinal hamartoma.

We isolated a single-cell-derived cell line from a spinal hamartoma, a which occurred in a newborn boy and was associated with a rudimentary limb. The maternal cells (HHC-7) differentiated into osteoblasts, chondrocytes, adipocytes, and skeletal muscles when they were cultured in differentiation-inducing media specific to each mesenchymal cell. We isolated a single-cell-derived clonal cell line (Clone K) after transfection with SV40 T antigen. These cells expressed CD73 and CD117, while being negative for expression of CD45. Clone K cells cultured in an osteogenic differentiation medium increased ALP activity and expressed mRNAs for Runx2 and osteocalcin. Treatment with rhBMP-2 induced Clone K cells to differentiate into both osteoblasts and chondrocytes. These cells expressed mRNAs for Sox9 and aggrecan in addition to osteogenic markers. Culture in an adipogenic differentiation medium induced Clone K cells to differentiation into adipocytes, which expressed mRNAs for PPARgamma2 and a2P. Clone K cells cultured in a serum-depleted medium generated desmin-positive cells and expressed MyoD1 mRNA. Clone K cells exhibited numerous alpha-smooth muscle actin-positive cells; however, treatment with rhBMP-2 decreased their number. Clone K cells, transplanted with a carrier containing rhBMP-2 into the muscles of SCID mice, generated ectopic endochondral bone formation. In these tissues, several osteoblasts and chondrocytes expressed SV40 T antigen, indicating their Clone K cell origin. Thus, Clone K cells are useful tools for analyzing the characteristics of human multipotential mesenchymal progenitors.

Doppler effect in resonant photoemission from SF6: correlation between Doppler profile and Auger emission anisotropy.

Fragmentation of the SF6 molecule upon F 1s excitation has been studied by resonant photoemission. The F atomiclike Auger line exhibits the characteristic Doppler profile that depends on the direction of the photoelectron momentum relative to the polarization vector of the radiation as well as on the photon energy. The measured Doppler profiles are analyzed by the model simulation that takes account of the anisotropy of the Auger emission in the molecular frame. The Auger anisotropy extracted from the data decreases with an increase in the F-SF5 internuclear distance.

Anisotropic ultrafast dissociation probed by the Doppler effect in resonant photoemission from CF4.

The resonant Auger spectrum from the decay of F 1s-excited CF4 is measured. Several lines exhibit a nondispersive kinetic energy as the exciting photon energy is tuned through the resonance region. The F 1s(-1) atomiclike Auger line is split into two components due to the emission of Auger electrons by a fragment in motion, when electron emission is observed along the polarization vector of the light. This Doppler splitting is direct evidence that the core excitation leads to T(d)-->C(3v) symmetry lowering, by elongation of a specific C-F bond preferentially aligned along the polarization vector of the incident photon.

Doppler-free resonant Raman auger spectroscopy of Ne+ 2s2p(5)3p excited states.

Using very high resolution achieved by the Doppler-free resonant Raman Auger technique, we have resolved the lowest terms of the series of inner-valence excitations 2s2p(5)((1,3)P)np 2S, 2P, and 2D in Ne+. The measured Auger anisotropic parameters and branching ratios help to establish the assignments of these levels. The measured lifetime widths are in reasonable agreements with ab initio calculations available in the literature.