Isoelectric focusing spectra of anti-bacterial alpha-amylase antibody unique for antigen-induced suppression.

The effect of intravenous (i.v.) administration of bacterial alpha-amylase (B alpha A) on the IgG antibody response to a subsequent challenge with B alpha A in incomplete Freund's adjuvant (IFA) varied with the difference in responsiveness of the parental strains. High-responder C3H/He (C3) mice given injections of either 200 or 4 micrograms of B alpha A, which alone were unable to trigger a detectable IgG antibody response, generated an enhanced response to an immunogenic challenge given 25 days after the last i.v. injection. The response of low-responder C57BL/6 (B6) mice previously exposed to B alpha A, following a different kinetic course depending on the exposing dose, reached a plateau lower than the levels of control responses (tentatively designated as high- and low-zone suppression). Prior exposure of (B6 X C3)F1 hybrids to 200 micrograms led to the enhanced response, whereas pretreatment with 4 micrograms rendered them partially tolerant to a subsequent challenge. These results suggest that the capacity to achieve low-zone suppression is inherited as a dominant trait. Isoelectric focusing (IEF) analysis revealed that these enhanced responses expanded antibody heterogeneity in a strictly restricted, strain-specific manner as observed during the normal antibody response, although the rate of expansion was accelerated. The specific antibodies produced by individual high-zone suppressed B6 mice were focused as a limited set of bands in a narrow pH range where the specific antibodies produced early in the normal response were focused. In contrast, the response of low-zone suppressed B6 and F1 hybrid mice was characterized by a unique process of heterogeneity expansion.     

Ingestion of High β-Glucan Barley Flour Enhances the Intestinal Immune System of Diet-Induced Obese Mice by Prebiotic Effects

The prebiotic effect of high β-glucan barley (HGB) flour on the innate immune system of high-fat model mice was investigated. C57BL/6J male mice were fed a high-fat diet supplemented with HGB flour for 90 days. Secretory immunoglobulin A (sIgA) in the cecum and serum were analyzed by enzyme-linked immunosorbent assays (ELISA). Real-time PCR was used to determine mRNA expression levels of pro- and anti-inflammatory cytokines such as interleukin (IL)-10 and IL-6 in the ileum as well as the composition of the microbiota in the cecum. Concentrations of short-chain fatty acids (SCFAs) and organic acids were analyzed by GC/MS. Concentrations of sIgA in the cecum and serum were increased in the HGB group compared to the control. Gene expression levels of IL-10 and polymeric immunoglobulin receptor (pIgR) significantly increased in the HGB group. HGB intake increased the bacterial count of microbiota, such as Bifidobacterium and Lactobacillus. Concentrations of propionate and lactate in the cecum were increased in the HGB group, and a positive correlation was found between these organic acids and the IL-10 expression level. Our findings showed that HGB flour enhanced immune function such as IgA secretion and IL-10 expression, even when the immune system was deteriorated by a high-fat diet. Moreover, we found that HGB flour modulated the gut microbiota, which increased the concentration of SCFAs, thereby stimulating the immune system.     

Total hip arthroplasty using hydroxyapatite-coated cementless cup for rapidly destructive coxarthrosis: Minimum 10-year follow-up

BackgroundPerforming total hip arthroplasty (THA) as early as possible is recommended for rapidly destructive coxarthrosis (RDC) as it causes pain that becomes progressively more severe. However, acetabular bone loss remains an issue in THA. Special devices, such as a Kerboull-type plate, may be used for acetabular bone defects, but the procedure is highly invasive and often the patients are elderly, further complicating matters. We retrospectively investigated the clinical and radiographic results of THA using conventional hydroxyapatite-coated cementless cup in RDC.MethodsA total of 32 patients (35 hips) with RDC were enrolled in the study with a minimum 10-year follow-up. All THAs were performed using conventional hydroxyapatite-coated cementless cup. All patients were evaluated clinically according to the Harris hip score (HHS). Acetabular bone deficiency was classified according to the American Academy of Orthopaedic Surgeons (AAOS) classification.ResultsEleven hips (31%) were AAOS type III, and none were type IV. Total HHS significantly improved from 36.5 to 79.4 (p < 0.01). Two cups exhibited loosening. The overall implant-associated survival rate after 10 years was 91.4%.ConclusionsClinical results of THA using conventional cementless implants for patients with RDC were acceptable. Thus, THA using conventional cementless implant is an effective and safe surgery for patients with RDC, minimizing surgical stress.     

Expression of membrane-bound and soluble receptor activator of NF-kappaB ligand (RANKL) in human T cells

The receptor activator of NF-kappaB ligand (RANKL) and its receptor RANK are critical regulators for immune responses as well as bone remodeling. RANKL is a type II transmembrane protein that has two forms-a membrane-anchored protein and a secreted protein. In this report, we demonstrate for the first time the kinetical expression of two forms of RANKL in human T cells using two monoclonal antibodies (mAbs) against human RANKL, which we newly derived. Freshly isolated T cells rarely expressed mRANKL, while the activation of T cells induced a substantial but minimal level of mRANKL as well as the accumulation of considerable amounts of sRANKL. The addition of the metalloprotease inhibitor KB-R8301 efficiently suppressed the release of sRANKL from activated T cells or RANKL-transfectants, and reciprocally enhanced the mRANKL expression. The membrane form of RANKL was also expressed on the infiltrating T cells in the rheumatoid synovial fluid and in the gingival tissues of patients with periodontitis. Our results demonstrate that the expression of mRANKL on T cells is strictly limited, and the majority of RANKL protein produced by T cells may be active in the soluble form after shedding. The mAbs that were derived in this study may be useful for investigating the regulation and function of RANKL in immune responses and bone remodeling.     

Alveolar soft part sarcoma, granular cell tumor, and paraganglioma. An immunohistochemical comparative study

Five cases of alveolar soft part sarcoma, 5 cases of granular cell tumor, and 6 cases of paraganglioma were investigated immunohistochemically to examine the expression of tissue-specific intermediate filaments (cytokeratin, vimentin, desmin, and glial fibrillary acidic protein (GFAP], actin, myoglobin, and nervous tissue markers (S-100 protein, neuron-specific enolase, and Leu-7). In alveolar soft part sarcomas, some of the tumor cells were positive for desmin, but negative for nervous tissue markers. The tumor cells of granular cell tumors were stained with anti-S-100 protein antibody, but not with anti-neuron-specific enolase antibody. In contrast, the tumor cells of paragangliomas were positive for neuron-specific enolase, but not for S-100 protein except for stellate cells surrounding the tumor cell nests. This immunohistochemical approach was valuable for the differential diagnosis of these three tumors. Furthermore, the complete absence of cytokeratin in all of the tumor cells may be helpful in distinguishing these three tumors from metastatic carcinoma in soft tissue. The histogenesis of alveolar soft part sarcoma is a matter of controversy. The result that besides desmin actin was also demonstrated in some of the tumor cells may support the myogenic origin of this tumor.     

Effects of GM1-ganglioside and α-sialyl cholesterol on amino acid uptake,protein synthesis,and Na+, K+-ATPase activity in superior cervical and nodose ganglia excised from adult rats

We examined the effect of GM₁-ganglioside in combination with cholera toxin B, and synthetic α-sialyl cholesterol (α-SC) on neutral amino acid (tritiated α-aminoisobutyric acid, [³H]AIB) uptake, protein synthesis ([³H]leucine incorporation), and Na⁺, K⁺-ATPase activity in isolated superior cervical ganglia (SCG) and nodose ganglia (NG) from adult rats after aerobic incubation, usually for 2 h at 37°C in vitro. Cholera toxin B, that specifically masks the oligosaccharide chain of GM₁-ganglioside, antagonized the GM₁-induced changes in [³H]AIB uptake, [³H]leucine incorporation, and Na⁺, K⁺-ATPase activity almost completely in SCG, but partially in NG. Although cholesterol itself had little effect on either [³H]AIB uptake and Na⁺, K⁺-ATPase activity both in SCG and NG, α-SC caused considerable reduction of both amino acid uptake and the transport enzyme activity in each ganglia. However, cholesterol was more effective than α-SC in decreasing [³H]leucine incorporation in either ganglia. Whereas addition of EGTA markedly reduced either GM₁-induced or α-SC-induced change in [³H]leucine incorporation into acid-insoluble fraction in both SCG and NG, application of Ca²⁺ ionophore produced considerable recovery of the protein synthesis from the inhibited level by Ca²⁺-deprivation. ATP and creatine phosphate contents in SCG were elevated by the presence of GM₁ or α-SC, whereas [³H]AIB uptake and Na⁺, K⁺-ATPase activity were inhibited, suggesting that utilization for membrane transport was diminished as a result of GM₁- or α-SC-induced decrease of ATPase activity.     

Retrovirus-mediated gene-transfer of tnf receptor for treatment of cancer

Tumor necrosis factor-alpha receptor cDNA (TNFR-1 or TNFR-2) was inserted into a retrovirus derived vector. Recombinant vector was transfected into packaging cell line. The transfectants producing high titer of virus containing TNFR-1 or TNFR-2 cDNA were obtained. Human stomach cancer cells that poorly expressed TNFR-1 or TNFR-2 were infected with this virus as an in vitro model experiment of gene therapy. The stomach cancer cells that were infected with recombinant virus expressed TNFR-1 or TNFR-2 and were effectively killed by recombinant TNF.     

Multiple tumor-suppressor-1 gene and esophageal-carcinoma

The multiple tumor suppressor 1 (MTS1) gene is homozygously deleted frequently in cell lines derived from a wide variety of tumors. We investigated the deletion of the MTS1 gene in esophageal cancer cell lines and primary esophageal squamous carcinomas using the polymerase chain reaction. Sixteen and 15 of 23 esophageal cancer cell lines showed homozygous deletion of MTS1 exon 1 and exon 2, respectively, while none of 21 primary esophageal carcinomas showed the deletion. An analysis of MTS1 gene mutations was carried out by direct DNA sequencing in 8 cell lines and 21 primary carcinomas showing no homozygous deletion. In contrast to previous reports of esophageal carcinoma, there were no mutations recognized in the region sequenced. Our study suggests that the inactivation of the MTS 1 gene may play an important role in esophageal carcinoma cell lines but may be less important in primary carcinomas of the human esophagus.     

MR imaging of the cochlear modiolus: area measurement in healthy subjects and in patients with a large endolymphatic duct and sac

PURPOSE: To evaluate the cochlear modiolus with thin-section magnetic resonance (MR) imaging in healthy subjects and patients with a large endolymphatic duct and sac, and to assess whether the cochlea is normal or abnormal in patients with a large endolymphatic duct and sac. MATERIALS AND METHODS: MR images were obtained in 10 ears in five volunteers (group 1), 40 ears in 20 patients with bilateral sensory hearing loss (group 2), three ears in two patients with Mondini malformation (group 3), and 12 ears in seven patients with a large endolymphatic duct and sac (group 4). RESULTS: In groups 1 and 2, all modiolar areas were larger than 4.0 mm2. In group 3, each modiolus was smaller than 2.0 mm2. In group 4, modiolar areas were smaller than 2.0 mm2 in eight ears and were larger than 4.0 mm2 in four ears. CONCLUSION: Findings in this study confirm that a large endolymphatic duct and sac is frequently associated with modiolar deficiency, but the modiolar area is normal in some cases. This result does not support the recently proposed hypothesis that hearing loss with a large endolymphatic duct and sac is caused by the transmission of subarachnoid pressure forces into the labyrinth through a deficient modiolus.