Discriminant equation using mucosally expressed cytokines and transcription factor for making definite diagnosis of inflammatory bowel disease unclassified.

BACKGROUND: The pathological conditions of UC and CD involved in inflammatory bowel disease-unclassified (IBD-U), UC with primary sclerosing cholangitis (PSC-UC), and UC with autoimmune pancreatitis type 2 (AIP-UC) remain unclear. Therefore, it is difficult to decide the appropriate treatments for these subtypes of UC. Our aim was to examine whether the discriminant equation using the mucosally expressed mediators designed as our previous study for IBD, could characterize IBD-U, PSC-UC, or AIP-UC. METHODS: A total of 56 patients including UC (n = 24), CD (n = 15), IBD-U (n = 10), PSC-UC (n = 4), and AIP-UC (n = 3), along with 9 control patients were enrolled in this study. Mucosally expressed inflammatory mediators related to Th1, Th2, Th17, and Treg were measured using quantitative PCR in endoscopic biopsies from the inflamed intestines of the patients. The IBD-U, PSC-UC or AIP-UC were characterized using discriminant analysis and principle component analysis. RESULTS: Through discriminant analyses, combinations of 3 to 7 inflammatory mediators were used to discriminate between UC and CD. Moreover, the identified 3 markers could diagnose patients with IBD-U as UC or CD with high accuracy. The distribution graph of inflammatory mediators using the principal component analysis revealed that PSC-UC and AIP-UC exhibited CD-like and UC-like features, respectively. CONCLUSIONS: The discriminant equation using mucosally expressed mediators of IL-13, IL-21 and T-bet can be used as a universal diagnostic tool not only for IBD-U but also to assess pathological conditions in PSC-UC and AIP-UC.

Interleukin-1ß as a Predictor of Glucocorticoid Response in Ulcerative Colitis.

BACKGROUND/AIM: Currently, there are no established biomarkers to differentiate between glucocorticoid (GC)-resistant and GC-sensitive ulcerative colitis (UC); however, interleukin (IL)-1ß could be one such candidate biomarker. The aim of this study was to investigate whether mucosally expressed IL-1ß could predict the response to GC in patients with UC. METHODS: A total of 27 mucosal tissue samples from 10 patients with GC-resistant UC (GC-resistant group), 9 patients with GC-sensitive UC (GC-sensitive group), and 8 control patients (control group) were analyzed by qRT-PCR for the expression of IL-1ß, GC receptor α (GRα), GRß, and other inflammatory mediators. Rachmilewitz endoscopic index (REI) between the GC-resistant and GC-sensitive groups was matched to avoid any potential influence of inflammation. RESULTS: The REI did not significantly differ between the GC-resistant and GC-sensitive groups. Mucosally expressed IL-1ß levels in the GC-resistant group were significantly higher than those in the GC-sensitive group. However, there were no significant differences in the expression levels of GRα, GRß, and other inflammatory mediators between the 2 groups. We could distinguish between the GC-resistant and GC-sensitive groups with a sensitivity of 90.0% and specificity of 77.8% based on mucosally expressed IL-1ß. CONCLUSIONS: Mucosally expressed IL-1ß can be used as a predictor of GC response in patients with UC.

Histological Features of Gastric Mucosa Serologically Diagnosed as Gastric Atrophy without Helicobacter pylori Infection.

BACKGROUND/AIMS: Although gastric atrophy is primarily caused by Helicobacter pylori infection, it is unclear why patients serologically diagnosed with gastric atrophy without H. pylori infection exhibit greater atrophy. We investigated histopathological features in serologically diagnosed gastric atrophy without H. pylori infection. METHODS: Thirty-four patients with positive serum pepsinogen and negative serum H. pylori antibody tests underwent gastric biopsy and histological evaluation. The presence of enterochromaffin-like cells (ECL) was also evaluated. Gastric cancer risks for each histological feature according to the Operative Link on Gastritis Assessment (OLGA) and Operative Link on Gastric Intestinal Metaplasia (OLGIM) were assessed. RESULTS: Twenty-five (74%) patients had histological gastric atrophy. Among those, the following histological subgroups were identified: eight had H. pylori but no ECL, 13 had neither H. pylori nor ECL, and 4 had ECL without H. pylori. Nine patients without histological atrophy had neither H. pylori nor ECL. Patients with H. pylori on histological diagnosis had significantly higher scores on OLGA and OLGIM. CONCLUSIONS: Various histological features, with significant differences in gastric cancer risk, were identified in the gastric mucosa serologically diagnosed with atrophy without H. pylori infection. Therefore, serological screening for gastric cancer risk tests has several limitations, and additional evaluations should be considered.

A practical regioselective synthesis of alkylthio- or arylthioindoles without the use of smelly compounds such as thiols.

A convenient method for the synthesis of 3-methylthioindoles has been established which does not use smelly compounds such as thiol derivatives. The method, which introduces an alkyl- or arylthio-group into the C(3)-position of the indole skeleton, was extended to the direct introduction of a methylthio or bromo group at the C(2)-position using 3-methylthioindoles. No dimerization occurred, and the reaction mechanism was confirmed. The products have the partial structure of potent anti-methicillin-resistant Staphylococcus aureus (anti-MRSA) bromomethylthioindoles (MC 5-8) isolated from marine algae. Furthermore, this reaction could be applied to the synthesis of 3,3-diindolyl thioether which is a core structure of Echinosulfone A.

Development of a new distyrylbenzene-derivative amyloid-ß-aggregation and fibril formation inhibitor.

Several new amyloid-ß (Aß) aggregation inhibitors were synthesized according to our theory that a hydrophilic moiety could be attached to the Aß-recognition unit for the purpose of preventing amyloid plaque formation. A distyrylbenzene-derivative, DSB(EEX)(3), which consider the Aß recognition unit (DSB, 1,4-distyrylbenzene) and expected to bind to amyloid fibrils (ß-sheet structure), was combined with the hydrophilic aggregation disrupting element (EEX) (E, Glu; X, 2-(2-(2-aminoethoxy)ethoxy)acetic acid). This DSB(EEX)(3) compound, compared to several others synthesized similarly, was found to be the most active for reducing Aß toxicity toward IMR-32 human neuroblastoma cells. Moreover, its inhibition of Aß-aggregation or fibril formation was directly confirmed by transmission electron microscopy and atomic force microscopy. These results suggest that the Aß aggregation inhibitor DSB(EEX)(3) disrupts clumps of Aß protein and is a likely candidate for drug development to treat Alzheimer's disease.

Development of an enzyme-linked immunosorbent assay system to determine the presence of antibodies specific for taxane structures.

Taxanes, which are widely used in treatment of numerous cancer types, are well-known to induce hypersensitivity reactions (HSR), especially in the case of paclitaxel. Although the cause of the HSR is commonly thought to be a non-immunological direct effect of the diluent which is used to dissolve paclitaxel, some reports suggest the possibility of the presence of an immunological reaction to the common taxane structure. The aim of this study was to establish a method to determine the presence of anti-taxane antibodies in body fluids of patients who have previously received paclitaxel, in order to estimate the risk of the occurrence of HSR to other taxane compounds, such as docetaxel. To prepare an enzyme-linked immunosorbent assay (ELISA) plate for determining taxanes, 10-deacetylbaccatin III (DAB) was first succinylated by use of dimethylaminopyridine and succinic anhydride in dried pyridine. After the succinylation reaction, three different products were obtained, and these were confirmed as 7-succinoyl DAB (7-DAB), 10-succinoyl DAB (10-DAB), and 7,10-disuccinoyl DAB (7,10-DAB) by (1)H-NMR analysis. Each of these three products was conjugated with bovine serum albumin (BSA), and adsorbed on an ELISA plate. By using a commercially available anti-taxane monoclonal antibody as a model antibody, the detection limit of the anti-taxane antibodies on the 7-DAB-BSA-, 10-DAB-BSA-, and 7,10-DAB-BSA-conjugated ELISA plate was estimated as 0.3, 1 and 10 ng/ml, respectively. The ELISA system established in this study may therefore be useful for estimating the risk of HSR to taxanes in a patient prior to the use of these drugs.

Posterior interosseous nerve palsy caused by synovial chondromatosis arising in the annular periradial recesses of the elbow.

We present a rare case report of a patient who presented with posterior interosseous nerve palsy caused by synovial chondromatosis. Synovial chondromatosis arising in the annular periradial recesses of the elbow joint was detected, and the mass developed two major portions constricted with the annular ligament. After surgical resection, posterior interosseous nerve palsy fully recovered and there was no recurrence of the lesion of synovial chondromatosis.

Role of the IL-23-T-bet/GATA3 Axis for the Pathogenesis of Ulcerative Colitis.

Ulcerative colitis (UC) has been considered a Th2- and Th17-related disease. However, anti-IL-12/23 p40 antibody, which blocks Th1 and Th17 cell induction and maintenance, has shown efficacy in treating UC, suggesting that UC might not be a prototypical Th2 and Th17 cell-mediated autoimmune disease. To verify how the immune responses in UC patients interact with each other, we analyzed the cytokine expression and transcription factors involved in the Th1, Th2, and Th17 responses. The mucosal expression of 19 cytokines and transcription factors related to Th1, Th2, and Th17 cells, as well as Tregs, were measured by quantitative polymerase chain reaction using endoscopic biopsy specimens from inflamed colons of UC patients. A correlation analysis between the cytokines and transcription factors was conducted. The characteristic cytokine profile in UC patients has two immune response clusters: Th17-related responses and Th1-/Th2-related responses. IL-23 showed a weaker association with Th17 cell-related cytokines and transcription factor RORC and a much stronger correlation with T-bet and GATA3. In the high-IL-23-expression group, the rate of chronic continuous type was higher and the remission rate lower than in the low-IL-23-expression group. IL-23 may be a very important cytokine for evaluating the UC disease condition, as the expression of IL-23 is associated with certain clinical characteristics of UC patients. A unique association between IL-23 and T-bet/GATA3 might play a key role in the pathogenesis of UC.

Mechanism for the direct synthesis of tryptophan from indole and serine: a useful NMR technique for the detection of a reactive intermediate in the reaction mixture.

The reaction mechanism for the biomimetic synthesis of tryptophan from indole and serine in the presence of Ac(2)O in AcOH was investigated. Although the time-course (1)H-NMR spectra of the reaction of 5-methoxyindole with N-acetylserine were measured in the presence of (CD(3)CO)(2)O in CD(3)CO(2)D, the reactive intermediate could not be detected. This reaction was conducted without 5-methoxyindole in order to elucidate the reactive intermediate, but the intermediate could not be isolated from the reaction mixture. Since the intermediate would be expected to have a very short life time, and therefore be very difficult to detect by conventional analytical methods, the structure of the intermediate was elucidated using a 2D-NMR technique, diffusion-ordered spectroscopy (DOSY). Two intermediates were detected and confirmed to be 2-methyl-4-methyleneoxazol-5(4H)-one and 2-methyl-4-hydroxymethyloxazol-5(4H)-one. The present results demonstrated that DOSY is a powerful tool for the detection of unstable intermediates.

Surgical treatment of stress fracture of the scaphoid of an adolescent gymnast.

Stress fracture of the carpal scaphoid is very rare. We present stress fracture of the scaphoid in an adolescent gymnast who was treated with internal fixation. Key pointsThere is a risk of stress fracture of the scaphoid in gymnast.Internal fixation with a screw is one of the good options for stress fracture of the scaphoid.

Subcutaneous Peroneus Longus Tendon Rupture Associated with OS Peroneum Fracture.

We report a rare case of subcutaneous peroneus longus tendon rupture associated with os peroneum fracture. Three dimensional computed tomographic scan was useful to understand this disorder. We treated the patient with excision of fractured os peroneum and tenodesis of the proximal stump of the ruptured peroneus longus tendon to the lateral aspect of the calcaneus. Key pointsIn order to understand a rare case of subcutaneous peroneus longus tendon rupture associated with os peroneum fracture, three dimensional computed tomographic scan was useful.The patient was treated with excision of fractured os peroneum and tenodesis of the proximal stump of the ruptured peroneus longus tendon to the lateral aspect of the calcaneus.

Familial Mediterranean Fever with Small Bowel Stenosis.

A 37-year-old man developed abdominal pain and the frequency of severe abdominal pain steadily increased to once a month. He was therefore admitted to our hospital. Abdominal CT showed bowel obstruction. It revealed transient stenosis in the small intestine. There were no symptoms such as fever or weight loss, it seemed unlikely that the patient had inflammatory bowel disease. Considering the history of recurrent abdominal pain, Familial Mediterranean Fever (FMF) was considered. As a result, a genetic analysis revealed mutations in exons 3 and 8 of the MEFV gene. We herein report the first known case of FMF with transient small bowel stenosis in Japan.

Endoscopic resection for residual lesion of metastatic gastric cancer: A case report.

BACKGROUND: Chemotherapy is a standard strategy for stage IV gastric cancer patients. However, some cases cannot undergo conversion surgery because of their frailty, even if the patients had response to chemotherapy. For these patients, local tumor progression is a problem. We report here the case of a patient whose residual gastric cancer was resected through endoscopic submucosal dissection (ESD) after concomitant chemotherapy for metastatic gastric cancer. CASE SUMMARY: An 85-year-old male complained of difficulty swallowing, and examination revealed gastric cancer with multiple liver metastases. Although he received concomitant chemotherapy, a residual tumor was observed in the primary lesion while the metastatic lesions disappeared completely. Conversion surgery was considered optional treatment; however, he could not undergo that because of advanced age and comorbidities. Thus, we performed ESD to treat the residual tumor. As a result, we resected the residual lesion completely. The patient has been alive for 29 mo since ESD, without recurrence. CONCLUSION: We achieved local control using ESD, and these findings may provide therapeutic improvements both in local control and patient survival outcomes.

Orally active factor Xa inhibitors: investigation of a novel series of 3-amidinophenylsulfonamide derivatives using an amidoxime prodrug strategy.

A series of novel and potent 3-amidinophenylsulfonamide derivatives of factor Xa inhibitors were designed and synthesized using an amidoxime prodrug strategy. We focused on systemic clearance of parent compounds in rats, and performed in vivo pharmacokinetic screening. Incorporation of a carboxymethoxy group markedly improved systemic clearance (compound 43), and the related amidoxime 44 showed sufficient prodrug conversion. Compound 45, the double prodrug of 43, exhibited practicable bioavailability after oral administration in rats. Among the various compounds under investigation, KFA-1982 was selected for clinical development.

Further structural analysis of GnRH complexes with metal ions.

MALDI-TOF mass spectrometry, 1H NMR spectrometry, the continuous variation method and molecular modeling by MM3 calculation confirmed our earlier studies showing that gonadotropin-releasing hormone (GnRH) forms complex with copper(II) ion with the binding ratio 1:1. The copper(II) complex formed at physiological pH has a square planar configuration and GnRH complexes with nickel(II) and cobalt(II) ions are less stable than that of copper(II).

Bone marrow metastatic myeloma cells promote osteoclastogenesis through RANKL on endothelial cells.

We have been using the B9/BM1 murine bone marrow metastasis model to study the function of adhesion molecules in the cell-cell interactions and transendothelial migration, necessary for tumor metastasis. The cell surface phenotype of these cells, which colonize vertebral and femoral marrow after intravenous injection, shows great similarity to that of human myeloma cells. In the present study, we investigated the interaction between B9/BM1 cells and osteoclasts, which likely support tumor metastasis in bone marrow. We found that co-culturing B9/BM1 cells and bone marrow-derived endothelial cells (BMECs) in the presence of vitamin D3 and M-CSF promoted differentiation of primary osteoclast progenitors to osteoclasts (detected by TRAP staining), and that this effect was blocked when BMECs were separated from the other cells by a porous polycarbonate membrane. Flow cytometry analysis showed that BMECs expressed RANKL (receptor activator of NF-kappaB ligand) protein on their surface, and that this expression was up-regulated by co-culture with B9/BM1 cells. Accordingly, RT-PCR showed expression of RANKL mRNA also to be up-regulated in BMECs co-cultured with B9/BM1 cells. Addition of OPG (osteoprotegerin, a decoy RANKL receptor) to the co-culture system completely blocked osteoclast induction, as did addition of anti-CD44 antibody. Furthermore, intravenous injection of B9/BM1 cells substantially increased the numbers of TRAP-positive osteoclasts detected in mice in vivo. Taken together, these findings suggest that B9/BM1 myeloma cells act via CD44 to stimulate RANKL expression on BMECs, which in turn physically interact with osteoclast progenitors to promote their differentiation to osteoclasts and metastasis in bone marrow.

Trigger lipids inducing pH-dependent liposome fusion.

Aspartic acid-derived artificial lipids (ADLs; s indicates the number of the methylene groups, s=2, 4, 6, 8, 10, 12) (Scheme 1) with various carboxyl alkyl chains as head groups are designed and synthesized, which are incorporated into liposome membranes by sonication. Fluorescence resonance energy transfer (FRET) measurements indicate that ADL6, ADL8 and ADL10 have high lipid-mixing ability in the acidic solution. The other ADLs, however, do not induce remarkable liposome fusion at acidic nor neutral pH. The hydrophobicity of the head groups of ADL6, ADL8 and ADL10 is suitable as triggers of membrane fusion.

Analysis of fibril formation of amyloid-beta-protein by stretched exponential function.

Kinetic behavior of aggregation of amyloid-beta-protein (Abeta) is represented by a stretched exponential function, F=A[1-exp(-Bt(n))]. Differences in temperature-dependence of A, B and n are studied for Abeta 1-40 and Abeta 1-42. As the temperature is lowered, parameter A is increased, parameter B is decreased and parameter n is increased in Abeta 1-40, while these parameters are less sensitive to temperature in a more hydrophobic protein Abeta 1-42. ln B is a linear function of n, which is shown by ln B = -6.34n + 3.69.

Evaluation of an interference screw for tendon reattachment to small bones.

OBJECTIVES: To compare the strength of interference screw fixation of the tendon to suture button fixation and staple fixation methods. DESIGN: Biomechanical study. METHODS: The femurs and flexor digitorum tendon of twelve rabbits were used. The tendon was attached to the medial condyle of the femur using one of the following three surgical procedures: (a) interference screw fixation: tendon-junction screw was used as an interference screw, (b) suture button fixation: the tendon was inserted through a hole and attached with a button on the opposite side, and (c) staple fixation: the tendon was attached using two staples. Strengths of the attached tendons were then measured using a universal testing machine. RESULTS: The ultimate failure load of interference screw fixation was 62.8 +/- 9.0 N, whereas that of suture button fixation was 13.9 +/- 3.8 N and staple fixation was 23.9 +/- 2.6 N. CONCLUSION: Interference screw fixation provides significant initial fixation strength (p < 0.05).

Polyarthritis caused by methimazole in two Japanese patients with graves' disease.

In many countries, methimazole (MMI) therapy is the first-line treatment in children with Graves' disease (GD). The rate of side effects of antithyroid drugs (ATDs) in children has been reported to range between 6% and 35%. Of these side effects, polyarthritis is uncommon but serious, and can also develop as a part of the antineutrophil cytoplasmic antibody-associated vasculitis that is induced by ATDs. Here, we describe two GD girl patients aged 15 years and 11 years who developed polyarthritis. The onset of polyarthritis in these patients was 24 days and 28 days after the initiation of MMI therapy, respectively. MMI was suspected of causing the polyarthritis in the two patients and was withdrawn. The symptoms of polyarthritis disappeared rapidly following cessation of treatment. Subsequently, one patient was treated with 131I therapy and the other patient was subjected to thyroidectomy. Although it rarely occurs in pediatric GD patients, severe polyarthritis is a serious side effect of MMI and is an indication for prompt cessation of treatment.