Total Synthesis of (+)-Silybin A.

We report the first total synthesis of silybin A (1). Key synthetic steps include the construction of the 1,4-benzodioxane neolignan skeleton, a modified Julia-Kocienski olefination reaction between m-nitrophenyltetrazole sulfone (m-NPT sulfone) 10 and aldehyde 21, the formation of the flavanol lignan skeleton 28 via a quinomethide intermediate under acidic conditions, and stepwise oxidation of the benzylic position of flavanol 29.

Acute Compartment Syndrome Following Reduction of Simple Elbow Dislocation.

Although rare, acute compartment syndrome may develop as a simple elbow dislocation after reduction without initial motor, sensory, or peripheral circulatory abnormalities. This report describes a rare case of this condition. Acute compartment syndrome remains a potential complication, even in a simple elbow dislocation without apparent initial abnormalities, and should be explained to patients. A peripheral nerve block during reduction may mask symptoms and delay recognition of acute compartment syndrome. This case highlights the importance of vigilant monitoring for acute compartment syndrome following reduction of simple elbow dislocations, especially when a peripheral nerve block is used during reduction.

An organic transistor for detecting the oxidation of an organic sulfur compound at a solid-liquid interface and its chemical sensing applications.

The development of chemical sensors has advanced due to an increase in demand; however, the potential of chemical sensors as devices to monitor organic reactions has not been revealed yet. Thus, we aim to propose a chemical sensor platform for facile monitoring of chemical reactions, especially at a solid-liquid interface. In this study, an extended-gate-type organic field-effect transistor (OFET) has been employed as a platform to detect chemical reactions at an interface between the extended-gate electrode and an aqueous solution. The OFET device functionalized with 4,4'-thiobisbenzenthiol has shown time- and concentration-dependent shifts in transistor characteristics upon adding H2O2. In a selectivity test using seven oxidant agents, the transistor responses depended on the oxidation of the organic sulfur compound (i.e., 4,4'-thiobisbenzenthiol) stemming from the ability of the oxidant agents. Therefore, the observed changes in the transistor characteristics have suggested the generation of sulfur-oxidized products at the interface. In this regard, the observed responses were caused by disulfide formation accompanied by changes in the charges under neutral pH conditions. Meanwhile, weak transistor responses derived from the generation of oxygen adducts have also been observed, which were caused by changes in the dipole moments. Indeed, the yields of the oxygen adducts have been revealed by X-ray photoelectron spectroscopy. The monitoring of gradual changes originating from the decrease in the disulfide formation and the increase in the oxygen adducts implied a novel aspect of the OFET device as a platform to simultaneously detect reversible and irreversible reactions at interfaces without using large-sized analytical instruments. Sulfur oxidation by H2O2 on the OFET device has been further applied to the indirect monitoring of an enzymatic reaction in solution. The OFET-based chemical sensor has shown continuous changes with an increase in a substance (i.e., lactate) in the presence of an enzyme (i.e., lactate oxidase), which indicates that the OFET response depends on the H2O2 generated through the enzymatic reaction in the solution. In this study, we have clarified the versatility of organic devices as platforms to monitor different chemical reactions using a single detection method.

1H-NMR-Based Metabolomics for the Classification of the Roots of Paeonia lactiflora, a Constituent of Kampo Medicines.

The root of Paeonia lactiflora (PAEONIAE RADIX) is a constituent of the traditional Japanese medicines (Kampo) and is known to have various effects. Peony roots cultivated in Japan and China are available in the Japanese market for medicinal use. In this study, the chemical diversity of ten available peony roots in the market that differed in their cultivation area was investigated using 1H-NMR metabolomics techniques. Principal component analysis and hierarchical cluster analysis of the 1H-NMR spectra of the peony roots methanolic extracts revealed a clear difference between the metabolic profiles of Japanese and Chinese peony roots. By preparative procedures using chromatography based on 1H-NMR spectra measurements, oxypaeoniflorin and (+)-catechin were found to be specific compounds for Japanese peony root. All peony roots used in this study were listed in the Japanese Pharmacopoeia. Therefore, the differences in the constituents of these peony roots might be attributed to growing conditions than differences in species. Cultivation conditions also influence the quality of natural medicines.

Gastric Invasive Micropapillary Carcinoma with Intestinal Phenotypes Harboring a TP53 R175H Mutation.

We report a case of gastric invasive micropapillary carcinoma (IMPC) in an 86-year-old female patient. She was admitted to our hospital with a chief complaint of bloody emesis. Upper gastrointestinal endoscopy found a gastric adenocarcinoma at the antrum. The biopsy specimens showed moderately differentiated adenocarcinoma with invasive small tumor nests. Distal gastrectomy with systematic lymph node dissection demonstrated that the tumor had IMPC through a pathological examination. Despite the depth of tumor invasion (the submucosa), extensive lymph node metastases were observed. Anti-D2-40 immunostaining revealed numerous infiltrating tumor cell nests in the lymphatic vessels, which could explain subsequent multiple lymph node metastases. The adenocarcinoma showed intestinal phenotypes by several immunohistochemical studies. One of these antibodies (CD10) clearly demonstrated the inverted apical-basal (inside-out) pattern of IMPC, whereas it showed an ordinary pattern in intestinal metaplasia adjacent to the tumor. Furthermore, genetic analysis by direct sequencing revealed a point mutation in the exon 5 of TP53 in the tumor. The mutation presumably harbors a missense mutation from Arg to His at the codon 175 (R175H). R175H has been previously described as a 'gain-of-function' mutation with a high invasive or metastatic potential in several types of cancers. In summary, this is one of the first reported cases of gastric IMPC with intestinal phenotypes harboring a TP53 R175H mutation in the literature.

Extended curettage and heat ablation for desmoplastic fibroma of the distal femur with a 12-year follow-up period: A case report.

Desmoplastic fibroma is a particularly rare, benign but locally aggressive, primary bone tumor. Owing to previously published reports stating high recurrence rates following curettage, the recommended primary treatment for desmoplastic fibroma is a marginal to wide tumor resection. In the current report, the case of an athlete with desmoplastic fibroma of the distal femur who was treated with extended curettage, heat ablation and artificial bone grafting is described. The postoperative course was uneventful and no recurrence has been observed during the 12-year follow-up period. The patient is able to sit on his heels with a straight back, without pain and is able run a complete marathon.

Combination of ultrasound and bubble liposome enhance the effect of doxorubicin and inhibit murine osteosarcoma growth.

If ultrasound (US) is applied to cells, permeability across the cell membrane temporarily increases, making it easier for drugs to be taken into the cells from around the cell membrane. Moreover, when used in combination with Bubble liposome (BL: liposomes which entrap an ultrasound imaging gas), even low-power ultrasound can facilitate drug delivery into cells. In the present study, we constructed a new drug delivery system (DDS) involving concomitant use of US and BL with doxorubicin (DOX), a key drug in the chemotherapy of osteosarcoma, and demonstrated both in vitro and in vivo that it markedly inhibited the proliferation of osteosarcoma cells. Furthermore, this system achieved an equivalent antitumor effect at about 1/5 the dose of antitumor agent employed in monotherapy with DOX. These findings suggest the possibility of reduction of adverse events. In this experiment, US and liposomes were tested, both of which are already in use in clinical practice. US and liposomes are both very safe in the body. The DDS composed of these elements we designed can be applied in simple and site-specific fashion and is therefore promising as a new, clinically feasible method of treatment.

Expression of active and inactive recombinant soluble trehalase using baculovirus-silkworm expression system and their glycan structures.

Silkworm soluble trehalase was expressed as a fusion protein with an N-terminal or C-terminal hexahistidine tag in a baculovirus-silkworm expression system and assayed for enzymatic activity. Only N-terminally tagged trehalase showed a high activity. This study is the first to report in vitro functional expression of recombinant insect soluble trehalase.

[A long term survivor of advanced gastric cancer treated with multi-drug combination chemotherapy].

We report a case of 70-year-old man who was admitted to our hospital due to hematemesis in June 2004. He was diagnosed by gastroscopy as having a type III moderately-poorly differentiated adenocarcinoma. A computed tomography (CT) scan revealed multiple lymph nodes swelling (#13, #16), finally he was diagnosed with gastric cancer stage IV (cT3, cN3, cM1). He was treated with S-1, but lymph nodes swelling increased in size, and then in March 2005, the treatment was changed to a second-line chemotherapy consisting of CPT-11 and CDDP. Abdominal CT scan showed a remarkable reduction of #16b1 lymph node, and the second-line chemotherapy was continued until 23 courses. But in April 2007, gastroscopy revealed the enlargement of gastric lesion. He was treated by third-line chemotherapy consisting of paclitaxel and doxifluoridine. This therapy was effective and continued until 7 courses. However, the treatment gradually became resistant and he died in May 2008, which was 4 years since the initial diagnosis.

Hyperbaric oxygen as a chemotherapy adjuvant in the treatment of osteosarcoma.

Although hyperbaric oxygen has been shown to enhance the efficacy of radiotherapy and chemotherapy for the treatment of several malignant tumors, the impact of hyperbaric oxygen on osteosarcoma has not yet been demonstrated. In this study, we investigated the efficacy of hyperbaric oxygen alone and in combination with an anti-cancer drug as an adjuvant to chemotherapy. In vitro, highly metastatic murine osteosarcoma cell lines were exposed to hyperbaric oxygen and cell viability was examined. Hyperbaric oxygen alone significantly suppressed cell proliferation, and hyperbaric oxygen plus carboplatin exhibited significant synergism in suppression of cell proliferation. In vivo, C3H mice were subcutaneously inoculated with osteosarcoma cells and divided into four groups: control, hyperbaric oxygen, carboplatin, and carboplatin plus hyperbaric oxygen. After 5 weeks, increase in both tumor volume and number of lung metastases was significantly suppressed in the hyperbaric oxygen group. Concomitant hyperbaric oxygen clearly enhanced the chemotherapeutic effects of carboplatin on both tumor growth and lung metastasis in osteosarcoma-bearing mice. Moreover, mortality in the carboplatin plus hyperbaric oxygen group was significantly lower than in the other three groups. These findings suggest that hyperbaric oxygen plus carboplatin combination therapy could be an appropriate therapeutic regimen for the treatment of patients with osteosarcoma.

Efficacy of the third-generation bisphosphonate risedronate alone and in combination with anticancer drugs against osteosarcoma cell lines.

BACKGROUND: Bisphosphonates (BPs) are currently the most important class of inhibitors of osteoclast-mediated bone resorption and are widely used in the treatment of osteoporosis and other osteoclast-mediated bone diseases. Recently, there has been increasing evidence that BPs also exhibit direct anti-tumor activity against several cancer cell lines. However, the efficacies of BPs against osteosarcoma have not yet been fully elucidated, and the anti-osteosarcoma activity of the potent new-generation BP risedronate, which is widely used clinically, has not been determined. MATERIALS AND METHODS: The anti-proliferative effects of the nitrogen-containing BP risedronate on seven osteosarcoma cell lines (LM8, SaOS2, U2OS, HOS, KHOS, MG63 and OST) were studied. The cell viability was determined by MTT assay. Prenylation of Rap1A and Ras and phosphorylation of Erk1 and 2 were examined by Western blot analysis. Genomic DNA fragmentation and TUNEL staining assay were performed to determine whether risedronate induced apoptosis of the osteosarcoma cells. The anti-tumor activities of risedronate in combination with carboplatin, doxorubicin, vincristine or etoposide were assayed with CalcuSyn (Biosoft). RESULTS: Risedronate inhibited both prenylation of Rap1A and Ras and phosphorylation of Erk 1 and 2 at a dose of 50 microM for 48 h. Treatment with risedronate induced significant time- and dose-dependent cytotoxicity in the LM8 cell line. Risedronate treatment also increased intranucleosomal genomic DNA fragmentation. The TUNEL assay showed that 10 microM and 50 microM risedronate clearly induced apoptosis of osteosarcoma cells. Risedronate also clearly inhibited LM8, SaOS2 and KHOS cell growth in a time- and dose-dependent fashion, but only weakly inhibited that of MG63 and U2OS cells. Risedronate augmented the effects of carboplatin, doxorubicin, vincristine and etoposide synergistically across a wide range of fractions affected (Fa) values. CONCLUSION: The nitrogen-containing bisphosphonate risedronate induces apoptosis and inhibits the growth of osteogenic sarcoma through a mechanism involving the down-regulation of protein prenylation and may be a candidate for combined use with carboplatin, doxorubicin, vincristine or etoposide.

Silkworm midgut proteins interacting with a hemolymph protease inhibitor, CI-8.

CI-8 is the chymotrypsin inhibitor in hemolymph from the silkworm, Bombyx mori. It occurs in the midgut at the spinning stage of larva, but little information on the mechanism of its uptake in the midgut is available. We found that two polypeptides interacting with CI-8 are in the midgut membrane, and we purified them using a biotinylated CI-8, viz., p29 and p60, having molecular sizes of 29 kDa and 60 kDa respectively. The structures of p29 and p60 were examined by N-terminal amino acid sequencing and peptide mass mapping, including tryptic digestion. p29 was highly similar to the matured 19G1-30K lipoprotein from hemolymph, but p60 was novel. Purified p29 was recognized by anti-19G1-30K antibody, and was confirmed to be similar to 19G1-30K. The antibody also neutralized the CI-8 binding ability of p29 in the midgut membrane. p29 and p60 are perhaps proteinaceous factors involved in the uptake of CI-8 into the midgut through the membrane.

Glucan-binding activity of silkworm 30-kDa apolipoprotein and its involvement in defense against fungal infection.

The silkworm Bombyx mori 30-kDa lipoproteins (6G1 and 19G1), major components of the hemolymph, were shown to bind to glucans. 6G1 apolipoprotein was expressed as a fusion protein with glutathione S-transferase in Escherichia coli and assayed for its binding activity. The purified recombinant 6G1 apolipoprotein specifically bound to beta-glucan, but not to chitin, mannan, peptidoglycan, or oligosaccharide chains on glycoproteins. The beta-glucan binding of the recombinant 6G1 was inhibited by laminaribiose and laminarin, a soluble glucan, but not by lipopolysaccharide or insect blood sugar, trehalose at physiological concentration. Furthermore, the recombinant 6G1 was shown to participate in the activation of prophenoloxidase cascade and to interfere with hyphal growth of the entomopathogenic fungus Paecilomyces tenuipes, injected into pupae of B. mori. These results suggest that 6G1 lipoprotein plays a role in the protection of B. mori against invading fungi.