Molecular allergen profiling in horses by microarray reveals Fag e 2 from buckwheat as a frequent sensitizer.

BACKGROUND: Companion animals are also affected by IgE-mediated allergies, but the eliciting molecules are largely unknown. We aimed at refining an allergen microarray to explore sensitization in horses and compare it to the human IgE reactivity profiles. METHODS: Custom-designed allergen microarray was produced on the basis of the ImmunoCAP ISAC technology containing 131 allergens. Sera from 51 horses derived from Europe or Japan were tested for specific IgE reactivity. The included horse patients were diagnosed for eczema due to insect bite hypersensitivity, chronic coughing, recurrent airway obstruction and urticaria or were clinically asymptomatic. RESULTS: Horses showed individual IgE-binding patterns irrespective of their health status, indicating sensitization. In contrast to European and Japanese human sensitization patterns, frequently recognized allergens were Aln g 1 from alder and Cyn d 1 from Bermuda grass, likely due to specific respiratory exposure around paddocks and near the ground. The most prevalent allergen for 72.5% of the tested horses (37/51) was the 2S-albumin Fag e 2 from buckwheat, which recently gained importance not only in human but also in horse diet. CONCLUSION: In line with the One Health concept, covering human health, animal health and environmental health, allergen microarrays provide novel information on the allergen sensitization patterns of the companion animals around us, which may form a basis for allergen-specific preventive and therapeutic concepts.

Eleostearic acid induces RIP1-mediated atypical apoptosis in a kinase-independent manner via ERK phosphorylation, ROS generation and mitochondrial dysfunction.

RIP1 is a serine/threonine kinase, which is involved in apoptosis and necroptosis. In apoptosis, caspase-8 and FADD have an important role. On the other hand, RIP3 is a key molecule in necroptosis. Recently, we reported that eleostearic acid (ESA) elicits caspase-3- and PARP-1-independent cell death, although ESA-treated cells mediate typical apoptotic morphology such as chromatin condensation, plasma membrane blebbing and apoptotic body formation. The activation of caspases, Bax and PARP-1, the cleavage of AIF and the phosphorylation of histone H2AX, all of which are characteristics of typical apoptosis, do not occur in ESA-treated cells. However, the underlying mechanism remains unclear. To clarify the signaling pathways in ESA-mediated apoptosis, we investigated the functions of RIP1, MEK, ERK, as well as AIF. Using an extensive study based on molecular biology, we identified the alternative role of RIP1 in ESA-mediated apoptosis. ESA mediates RIP1-dependent apoptosis in a kinase independent manner. ESA activates serine/threonine phosphatases such as calcineurin, which induces RIP1 dephosphorylation, thereby ERK pathway is activated. Consequently, localization of AIF and ERK in the nucleus, ROS generation and ATP reduction in mitochondria are induced to disrupt mitochondrial cristae, which leads to cell death. Necrostatin (Nec)-1 blocked MEK/ERK phosphorylation and ESA-mediated apoptosis. Nec-1 inactive form (Nec1i) also impaired ESA-mediated apoptosis. Nec1 blocked the interaction of MEK with ERK upon ESA stimulation. Together, these findings provide a new finding that ERK and kinase-independent RIP1 proteins are implicated in atypical ESA-mediated apoptosis.

Sensitization to acid-hydrolyzed wheat protein by transdermal administration to BALB/c mice, and comparison with gluten.

BACKGROUND: An increasing number of studies have shown that hydrolyzed wheat protein (HWP) can induce IgE-mediated hypersensitivity by skin contact and/or food ingestion. However, there has been no study of the sensitizing potential of HWP. In this study, the possibility of transdermal pathway for sensitization to acid-HWP (HWP1) was investigated using BALB/c mice, and compared with that of gluten. METHODS: HWP1 or gluten (500 µg/mouse) was transdermally administered using patches. After three or four cycles of sensitization for 3 days/week, active systemic anaphylaxis (ASA) was induced by intraperitoneal injection of the antigen, and rectal temperatures, scores of anaphylactic responses, and plasma histamine levels were determined. Because HWP1 was included in facial soap in Japan, the effect of detergent on the sensitizing potential was also investigated. RESULTS: Transdermal administration of HWP1 induced dose-dependent production of IgE and IgG1. After sensitization for 3 or 4 weeks, intraperitoneal injection of HWP1 caused ASA, leading to decreased rectal temperatures, increased anaphylaxis scores, and increased plasma histamine levels. In addition, splenocytes harvested after ASA produced IL-4, IL-5, and IL-10 by re-stimulation with HWP1. Transdermal exposure to gluten also induced IgE and IgG1 production, and intraperitoneal injection of gluten also induced ASA only in mice sensitized in the presence of sodium dodecyl sulfate. CONCLUSIONS: Transdermal exposure to HWP1 is sufficient to activate key immune pathways necessary for sensitizing mice for immediate hypersensitivity reactions. This study shows that HWP has a sensitizing potential as well as gluten, whereas its allergenicity may be different from that of gluten.

Posterior occipitocervical fixation under skull-femoral traction for the treatment of basilar impression in a child with Klippel-Feil syndrome.

We present the case of a 15-year-old boy with symptoms due to Klippel-Feil syndrome. Radiographs and CT scans demonstrated basilar impression, occipitalisation of C1 and fusion of C2/C3. MRI showed ventral compression of the medullocervical junction. Skull traction was undertaken pre-operatively to determine whether the basilar impression could be safely reduced. During traction, the C3/C4 junction migrated 12 mm caudally and spasticity resolved. Peri-operative skull-femoral traction enabled posterior occipitocervical fixation without decompression. Following surgery, cervical alignment was restored and spasticity remained absent. One year after surgery he was not limited in his activities. The surgical strategy for patients with basilar impression and congenital anomalies remains controversial. The anterior approach with decompression is often recommended for patients with ventral compression of the medullocervical region, but such procedures are technically demanding and carry a significant risk of complications. Our surgical strategy was an alternative solution. Prior to a posterior cervical fixation, without decompression, skull traction was used to confirm that the deformity was reducible and effective in resolving associated myelopathy.

A convenient and sensitive allergy test: IgE crosslinking-induced luciferase expression in cultured mast cells.

BACKGROUND: For the detection of allergen-specific IgE in sera, solid-phase IgE-binding assays like the CAP test are commonly used. Although such immunochemical methods are very sensitive, they frequently produce false positives. Degranulation of the human IgE receptor (FcεRI)-transfected rat mast cell (RBL) lines seems to be a possible indicator for human IgE, but spontaneous mediator release from these cells in the presence of human sera is not negligible. METHODS: The nuclear factor of activated T-cells (NFAT)-responsive luciferase reporter gene was stably transfected into human FcεRI-expressing RBL-SX38 cells. One established clone (RS-ATL8) was sensitized with 1 : 100 dilution of sera from patients with egg white allergy and then stimulated with purified or a crude extract of egg white allergen. RESULTS: Sensitization with 15 pg/ml IgE was sufficient to detect IgE crosslinking-induced luciferase expression (EXiLE) by anti-IgE stimulation. Allergen-specific EXiLE was elicited by as little as 1 fg/ml of egg white protein without cytotoxicity. There was a good correlation between results with EXiLE and oral food challenge tests on patients with egg allergy (P = 0.001687, Fisher's exact test). The measured values of EXiLE and the CAP test also correlated well (R = 0.9127, Spearman's test). CONCLUSION: The EXiLE test using RS-ATL8 cells is a promising in vitro IgE test to evaluate the biological activity of the binding between IgE and allergens.

Severe rotational deformity, quadriparesis and respiratory embarrassment due to osteomyelitis at the occipito-atlantoaxial junction.

We present the case of an 83-year-old man who developed quadriparesis and respiratory embarrassment following osteomyelitis at the occipito-atlantoaxial junction. He had developed an abscess at this site after an earlier urinary infection with methicillin-resistant staphylococcus aureus. Stabilisation of the neck and antibiotic therapy led to an almost complete neurological recovery without recourse to anterior surgery.

Incidence and risk factors of falling in ambulatory patients with rheumatoid arthritis: a prospective 1-year study.

UNLABELLED: A prospective 1-year study showed that fall incidence was 50% in women with rheumatoid arthritis. Multivariate analysis identified swollen joint count, use of antihypertensives or diuretics, one-leg standing time, and sway area measured by stabilometer as significant parameters associated with falls. INTRODUCTION: Patients with rheumatoid arthritis (RA) may be at increased risk of falling because they frequently experience muscle weakness and stiff or painful joints. The aim of this study was to use a prospective design to determine the incidence of falls and their risk factors in women with RA. METHODS: Eighty-four women aged 50 and over who had RA were enrolled. The mean age was 64.1 years. We evaluated postural stability, physical performance related to falls, disease activity, muscle volume, and bone density. The occurrence of falls was assessed every month for 1 year. Among 84 patients, 80 completed a 1-year observation. RESULTS: Forty patients (50.0%) reported one or more falls, and two of them (5.0%) had fractures during the follow-up period. The fall group had more swollen joints and took more antihypertensives and/or diuretics. The fall group also had lower postural stability and tended to have reduced physical performance. The one-leg standing time was shorter, and the step-up-and-down test score was lower in the fall group. The sway area was larger in the fall group. DISCUSSION: Multiple logistic regression analysis identified that number of swollen joints, use of antihypertensives or diuretics, shorter time standing on one foot, and the sway area were the most significant parameters associated with falls. CONCLUSION: We concluded that fall rates in RA patients were higher than in the general population and that balance impairment or side effects of drugs may play a role in increasing the risk of falls.

Sequential change in quality of life for patients with incident clinical fractures: a prospective study.

UNLABELLED: Health-related quality of life in elderly women with sustained incident fractures was assessed prospectively for 1 year, using the EuroQol standard. Loss of QOL was more severe in patients after hip or vertebral fractures than those with wrist fracture. QOL was not completely restored in patients suffering from hip fracture. INTRODUCTION: Osteoporosis-related fractures decrease mobility, social interaction, and emotional well-being. All of these characteristics determine health-related quality of life (HR-QOL). In this study, we assessed HR-QOL in elderly women following incident clinical fractures. METHODS: Thirty-seven patients with hip fractures (mean age 76.1 years), 35 with vertebral fractures (mean age 72.6 years), and 50 with wrist fractures (mean age 68.6 years) were enrolled. HR-QOL was prospectively measured using EuroQol (EQ-5D) before the fracture, 2 weeks, 3 months, 6 months, and 1 year after the fracture. RESULTS: During the observation period, reduction of EQ-5D values was greatest in the hip fracture group. In the wrist fracture group, EQ-5D values at 6 months after the fracture showed recovery; however, in the hip and vertebral fracture groups, recovery was significantly lower than before the fracture. One year after the fracture, EQ-5D values were not significantly different from prefracture values in the vertebral and wrist fracture groups, but remained significantly lower in the hip fracture group. CONCLUSIONS: Loss of QOL was more severe in patients after hip or vertebral fractures than in patients with wrist fracture. HR-QOL was not completely restored in patients suffering from hip fracture.

Recent trends in the incidence and lifetime risk of hip fracture in Tottori, Japan.

SUMMARY: Hip fracture incidence from 2004 to 2006 in the Tottori prefecture of Japan was investigated and compared with previously reported rates. The age- and gender-specific incidence of hip fracture in the Tottori prefecture has not plateaued, as has been reported for populations in Northern Europe or North America. INTRODUCTION: Recent data from Northern Europe and North America indicate that the incidence of hip fracture has plateaued, whereas most reports from Asia indicate that the incidence is increasing. The aims of this study were to investigate the recent incidence of hip fracture in the Tottori prefecture, Japan, and to compare it with previous reports. METHODS: All hip fractures in patients aged 35 years and older occurring between 2004 and 2006 were surveyed in all of the hospitals from the Tottori prefecture. The age- and gender-specific incidence rates were then calculated. Using these and previously reported data, the estimated number of hip fracture patients was determined using the age- and gender-specific incidence rates in each year from 1986 to 2006. RESULTS: The survey identified 851, 906, and 1,059 patients aged 35 years and older, in 2004, 2005, and 2006 respectively. The residual lifetime risk of hip fracture for individuals at 50 years of age was estimated to be 5.6% for men and 20.0% for women. The estimated number of patients from 1986 to 2006 showed a significant increase over time for both genders. CONCLUSIONS: The age- and gender-specific incidence of hip fracture in the Tottori prefecture, Japan has not plateaued for either gender.

Measurement of human trabecular bone by novel ultrasonic bone densitometry based on fast and slow waves.

SUMMARY: Two longitudinal transmitted waves, fast and slow waves, were observed by employing a new quantitative ultrasound (QUS) method. The trabecular bone measurements generated by this method reflect three-dimensional structural information, and the new QUS parameters were able to identify vertebral fractures. INTRODUCTION: The aims were to identify new quantitative ultrasound (QUS) parameters that based on new QUS method reflecting not only bone volume but also the microstructures of trabecular bone ex vivo and to observe how much they predict fracture risk in vivo. METHODS: Ex vivo measurement: Three human femoral heads were used for the experiment. Attenuation of the slow wave, attenuation of the fast wave, speed of the slow wave, speed of the fast wave (SOFW), bone mass density of trabecular bone, and elastic modulus of the trabecular bone (EMTb) of each specimen were obtained using a new QUS method and compared with three-dimensional structural parameters measured by micro-computed tomography. In vivo measurement: Eighty-nine volunteers were enrolled, and the bone status in the distal radius was measured using a new QUS method. These parameters were compared with data evaluated by peripheral quantitative computed tomography and dual X-ray absorptiometry. RESULTS: Ex vivo measurement: SOFW and EMTb showed correlations with the parameter of trabecular anisotropy. In vivo measurement: The new QUS parameters were able to identify vertebral fractures. CONCLUSION: The newly developed QUS technique reflects the three-dimensional structure and is a promising method to evaluate fracture risk.

Effect of a selective agonist for prostaglandin E receptor subtype EP4 (ONO-4819) on the cortical bone response to mechanical loading.

The influence of a selective agonist for prostaglandin E receptor subtype EP4 (ONO-4819) on the bone response to mechanical loading was evaluated. Six-month-old female Wistar rats were used and assigned to three groups (n = 12/group): Vehicle administration (EP4-V), low-dose ONO-4819 administration (EP4-L, 3 microg/kg BW), and high-dose ONO-4819 administration (EP4-H, 30 microg/kg BW). ONO-4819 was subcutaneously injected in the back twice a day for 3 weeks. Loads on the right tibia at 39.4 N for 36 cycles at 2 Hz were applied in vivo by 4-point bending every other day for 3 weeks. Whole-body bone mineral content showed a significant difference between EP4-V and EP4-H (P < 0.05). Bone mineral density (BMD) of the total and regional tibia (the region with maximal bending at the central diaphysis) was higher in EP4-H than EP4-V, showing a significant effect of loading (P < 0.001) and ONO-4819 (P < 0.05). BMD of the total femur was higher in EP4-H than EP4-V (P < 0.01) and that of the distal femur was higher in EP4-H than EP4-V (P < 0.001). Histomorphometry of the cortical bone showed that loading increased formation surface (FS/BS), mineral appositional rate (MAR), and bone formation rate (BFR/BS) significantly at the lateral periosteal surface (P < 0.001); however, the effect of ONO-4819 was not significant. At the medial periosteal surface, loading increased the three parameters (P < 0.001) and ONO-4819 increased FS/BS (P < 0.001) and MAR (P < 0.05) significantly. At the endocortical surface, the effects of both loading and ONO-4819 were significant on all three parameters (for loading; FS/BS P < 0.01, MAR P < 0.05, BFR/BS P < 0.03, for ONO-4819 all P < 0.001). It was concluded that ONO-4819 increased cortical bone formation in rats and there was an additive effect on the bone response to external loading by 4-point bending.

Gene expression relevant to osteoclastogenesis in the synovium and bone marrow of mature rats with collagen-induced arthritis.

OBJECTIVES: To investigate gene expression relevant to osteoclastogenesis in the synovium and bone marrow during the development of collagen-induced arthritis (CIA) in mature rats. METHODS: Total messenger RNAs (mRNAs) were obtained from CIA synovium and bone marrow after immunization. First, reverse transcriptase-polymerase chain reactions (RT-PCR) were carried out to detect the mRNA encoding receptor activator of NF-kappaB (RANK), RANK ligand (RANKL), osteoprotegerin (OPG), tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-6 and the osteoclast markers tartrate-resistant acid phosphatase (TRAP) and cathepsin K. Secondly, the genes detected clearly by RT-PCR were quantified using real-time PCR. RESULTS: In the synovium, expression of all genes was confirmed by specific single bands in RT-PCR. In real-time PCR, the expression levels of TNF-alpha, IL-1beta, IL-6, RANKL, TRAP and cathepsin K mRNA increased, whereas the expression levels of RANK and OPG were unchanged and decreased respectively. RANKL expression was highly correlated with the two osteoclast markers. In the bone marrow, RT-PCR did not clearly detect the expression of IL-6, RANKL or OPG mRNA. Quantitative real-time PCR showed that TNF-alpha, RANK and TRAP mRNA expression did not change significantly with time, and that IL-1beta and cathepsin K changed slightly compared with those in the synovium. CONCLUSIONS: In the early stages of arthritis, synovial RANKL is closely involved in osteoclastogenesis, and various changes in synovial cytokines, including down-regulation of OPG, probably accelerate osteoclast formation. In contrast, cytokine mRNA in the bone marrow showed little fluctuation. We suggest that synovial cytokines affect osteoclastogenesis not only in the synovium but in the bone marrow.

A multi-laboratory evaluation of a common in vitro pepsin digestion assay protocol used in assessing the safety of novel proteins.

Rationale. Evaluation of the potential allergenicity of proteins derived from genetically modified foods has involved a weight of evidence approach that incorporates an evaluation of protein digestibility in pepsin. Currently, there is no standardized protocol to assess the digestibility of proteins using simulated gastric fluid. Potential variations in assay parameters include: pH, pepsin purity, pepsin to target protein ratio, target protein purity, and method of detection. The objective was to assess the digestibility of a common set of proteins in nine independent laboratories to determine the reproducibility of the assay when performed using a common protocol. Methods. A single lot of each test protein and pepsin was obtained and distributed to each laboratory. The test proteins consisted of Ara h 2 (a peanut conglutin-like protein), beta-lactoglobulin, bovine serum albumin, concanavalin A, horseradish peroxidase, ovalbumin, ovomucoid, phosphinothricin acetyltransferase, ribulose diphosphate carboxylase, and soybean trypsin inhibitor. A ratio of 10U of pepsin activity/microg test protein was selected for all tests (3:1 pepsin to protein, w:w). Digestions were performed at pH 1.2 and 2.0, with sampling at 0.5, 2, 5, 10, 20, 30, and 60min. Protein digestibility was assessed from stained gels following SDS-PAGE of digestion samples and controls. Results. Results were relatively consistent across laboratories for the full-length proteins. The identification of proteolytic fragments was less consistent, being affected by different fixation and staining methods. Overall, assay pH did not influence the time to disappearance of the full-length protein or protein fragments, however, results across laboratories were more consistent at pH 1.2 (91% agreement) than pH 2.0 (77%). Conclusions. These data demonstrate that this common protocol for evaluating the in vitro digestibility of proteins is reproducible and yields consistent results when performed using the same proteins at different laboratories.

Case-control study of risk factors for fractures of the distal radius and proximal humerus among the Japanese population.

We conducted a case-control study to identify risk factors for fractures of the distal radius and proximal humerus. Subjects were selected from women aged 45 and over with distal radius and proximal humerus fractures, resulting from minor trauma. Two age- and gender-matched controls for each case were selected from patients who subsequently visited the same clinic for treatment of conditions other than fractures. Questionnaires including anthropometric data, past and current physical activity, and lifestyle were sent by mail to both subjects and controls. A total of 140 women with distal radius fractures (mean age 67.4 years) and 242 controls were analyzed. Falls during the previous year were a significant risk factor, while futon use (instead of bed use) before fracture was a protective factor for distal radius fractures. A total of 37 women with proximal humerus fractures (mean age 76.3 years) and 67 controls were analyzed. Weight loss was a significant risk factor, while greater frequency of going outside significantly decreased the risk of proximal humerus fracture. There was no significant correlation with eating habits, milk and alcohol consumption, or smoking to the risk of either fracture.

Nerve-mast cell (RBL) interaction: RBL membrane ruffling occurs at the contact site with an activated neurite.

Mast cell-neurite interaction serves as a model for neuroimmune interaction. We have shown that neurite-mast cell communication can occur via substance P interacting with neurokinin (NK)-1 receptors on the mucosal mast cell-like cell, the rat basophilic leukemia (RBL) cell. Neurite (murine superior cervical ganglia) and RBL cell [expressing the granule-associated antigen CD63-green fluorescent protein (GFP) conjugate] cocultures were established and stimulated with bradykinin (BK; 10 nM) or scorpion venom (SV; 10 pg/ml), both of which activate only neurites. Cell activation was assessed by confocal imaging of Ca2+ (cells preloaded with fluo 3), and analyses of RBL CD63-GFP+ granule movement were conducted. Neurite activation by BK or SV was followed by RBL Ca2+ mobilization, which was inhibited by an NK-1 receptor antagonist (NK-1 RA). Moreover, membrane ruffling was observed on RBL pseudopodial extensions in contact with the activated neurite, but not on noncontacting pseudopodia. RBL membrane ruffling was inhibited by NK-1 RA, but not NK-2 RA, and was accompanied by a significant increase in granule movement (0.13 +/- 0.04 vs. 0.05 +/- 0.01 microm/s) that was most evident at the point of neurite contact: many of the granules moved toward the plasmalemma. This is the first documentation of such precise (restricted to the membrane's contact site) transfer of information between nerves and mast cells that could allow for very subtle in vivo communication between these two cell types.

Cross-sectional and longitudinal study of osteoporosis in patients with rheumatoid arthritis.

To elucidate the pathology of osteoporosis associated with rheumatoid arthritis (RA), bone mass measurements were performed in 146 female patients with RA and compared with those in 150 age-matched female patients with osteoarthritis (OA) and postmenopausal osteoporosis (OP). Bone mineral density (BMD) was measured at the lumbar spine (L-BMD), the mid-radius (MR-BMD) and the calcaneus (C-BMD) by dual-energy X-ray absorptiometry (DXA), and at the distal radius by peripheral quantitative computed tomography (pQCT). The RA group showed significantly lower BMD at all sites, except L-BMD, than the OA group. Compared with the OP group, the RA group showed a significantly higher L-BMD but no difference at other sites. BMD in RA decreased with disease severity at all sites and lean body mass was highly correlated with L-BMD and C-BMD. Cross-sectional analysis revealed early bone loss at the distal radius and a decrease of L-BMD, MR-BMD, and C-BMD with disease duration. Longitudinal analysis showed that the annual loss of L-BMD, MR-BMD and C-BMD tended to be lower with increasing disease duration. Glucocorticoid administration had no influence on L-BMD, MR-BMD or C-BMD. We concluded that, unlike postmenopausal osteoporosis, osteoporosis associated with RA is characterised by relatively preserved bone mass in the axial bone and marked loss in the peripheral bone. The risk factors for generalised osteoporosis are a long disease duration, severity of disease, and decreased lean body mass.

Re-aggravation of myelopathy due to intramedullary lesion with spinal cord enlargement after posterior decompression for cervical spondylotic myelopathy: serial magnetic resonance evaluation.

STUDY DESIGN: A case report. OBJECTIVES: To report a case of swelling of the spinal cord and an intramedullary lesion occurring after expansive laminoplasty for cervical spondylotic myelopathy. SETTING: A university hospital in Japan. METHODS: Clinical evaluation, radiography, MR imaging. RESULTS: A 65-year-old man with a cervical spondylotic myelopathy in whom symptoms were improved immediately after expansive laminoplasty, but became aggravated 2 weeks later. Magnetic resonance images demonstrated swelling of the spinal cord and an intramedullary lesion that extended from the medulla oblongata to C7. Nine months after surgery, the lesion was reduced to C2-6, but neurological deterioration had not improved. Six years after surgery, the patient remains confined to bed. CONCLUSION: Patients with such disease conditions are rare, and it is difficult to predict postoperative swelling of the spinal cord before surgery. Spine surgeons should be aware of such rare disease conditions involving the spinal cord.

Measurement of periprosthetic bone mineral density by dual-energy x-ray absorptiometry is useful for estimating fixation between the bone and the prosthesis in an early stage.

Periprosthetic bone mineral density (BMD) was measured with dual-energy x-ray absorptiometry in 37 hips after cementless hip arthroplasty. Changes in BMD were evaluated longitudinally from 1 to 36 months. Stem fixation was evaluated at 5 years after surgery and was classified into a stable fixation group (30 hips) and an unstable fixation group (7 hips). On plain radiographs, distinct reactive lines around the proximal stem were detected at an average of 35 months after surgery in the unstable fixation group. In the unstable fixation group, the BMD decrease in the calcar region peaked 6 months after surgery, and BMD increased thereafter. In the stable fixation group, BMD continued to decrease until 36 months after surgery; a significant difference was observed between the 2 groups 24 months after surgery. Measurement of periprosthetic BMD is useful for estimating fixation between the bone and the prosthesis in an early stage.

Gene expression profiling of Ca2+-atpase inhibitor DTBHQ and antigen-stimulated RBL-2H3 mast cells.

OBJECTIVE AND DESIGN: Ca2+ signaling is critical for mast cell activation by antigen stimulation, and we previously described that the signaling can be mimicked by Ca2+-ATPase inhibitors. We therefore investigated the effect of the Ca2+-ATPase inhibitor and antigen stimulation on the gene expression profiles of RBL-2H3 mast cells. MATERIAL: A Ca2+-ATPase inhibitor, 2,5-di(tert-butyl)-1,4-hydroquinone (DTBHQ), an antigen (dinitrophenylated BSA), a high-density oligonucleotide microarray (Affymetrix GeneChip) technique, and a well-characterized rat mast cell line RBL-2H3 were used. TREATMENT: RBL-2H3 cells were activated for 3 h with 10 microM DTBHQ, which increases cytosolic Ca2+ concentration, or 10 microg/ml antigen, which cross-links IgE receptors, and the mRNA expression profiles (8,799 genes) were analyzed with GeneChip arrays (n = 3). METHODS: Expression levels were measured by GeneChip, and the differences were tested by Welch's t-test and P-values less than 0.05 were considered statistically significant. Values are expressed as means +/- SEM. RESULTS: The genes, including MCP-1, GADD45, Relaxin H1, CSF-1, c-jun-oncogene, Pyk-2, NKR-P2 and CREM, were significantly up-regulated by both DTBHQ and antigen stimuli, whereas the genes including interleukin (IL)-3, IL-4, IL-9, IL-13, GADD153, butyrate response factor, and Fas ligand, were up-regulated by DTBHQ alone. On the other hand, the expression of several genes, including GATA-1, were down-regulated by DTBHQ stimulation. CONCLUSIONS: These results suggest 1) that DTBHQ seems to induce proinflammatory responses by stimulating the production of several cytokines through the expression of several transcription factors, 2) that the changes in gene expression profile induced by DTBHQ and by IgE receptor cross-linking in mast cells were almost the same, but many more stress-inducible genes like GADD 153 were up-regulated by the former.

1H-NMR studies of cerebrospinal fluid: endogenous ethanol in patients with cervical myelopathy.

Endogenous ethanol was observed by nuclear magnetic resonance spectroscopy in the course of screening for cerebrospinal fluid of the patients with cervical myelopathy. Ethanol was detected in 10 out of 20 patients. It seems likely that the presence of endogenous ethanol is related to the severity of myelopathy. Also, the concentration of ethanol was correlated with that of lactate in the cerebrospinal fluid. This implies that ethanol may be formed as the end product of glycolysis or in an unknown pathway in the case of severely insulted myelonic tissues.