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Epithelioid angiomyolipoma has malignant potential; however, no effective therapy has been established for advanced cases. A 50-year-old woman with a history of right nephrectomy for epithelioid angiomyolipoma was referred to our institution. Computed tomography and magnetic resonance imaging showed multiple tumors in her lung, liver and pelvic cavity. The liver and pelvic tumor specimens obtained by needle biopsy confirmed the diagnosis of epithelioid angiomyolipoma recurrence. The patient was treated with everolimus (10 mg/day). Three months later, pulmonary lesions disappeared; liver and pelvic tumors significantly shrank in size, but the pelvic tumor gradually enlarged again. We carried out surgical resection of the residual liver and pelvic cavity tumors. Although the mammalian target of rapamycin inhibitor seems to be effective for treating epithelioid angiomyolipoma, its long-term effects remain unknown. Thus, aggressive administration of a multidisciplinary treatment including molecular target therapy and surgical resection is required to improve the prognosis of epithelioid angiomyolipoma., (© 2013 The Japanese Urological Association.)

We report three patients who were successfully treated with orthopedic surgery for spinal paralysis due to spine metastasis of gastric cancer. Case 1: A 75-year-old female had lower limb paralysis due to metastasis of Th1-Th3 one year and four months after total gastrectomy for gastric cancer. Although radiotherapy was carried out from the day following a paralysis appearance, the paralysis deteriorated from D to C. Then, when direct decompressive surgical resection had been performed eight days after the paralysis appearance, the paralysis improved from C to D. Case 2: A 50-year-old male had lower limb paralysis due to metastasis of Th2 three years and eight months after distal gastrectomy for gastric cancer. Paralysis had improved from C to D with direct decompressive surgical resection three days after the paralysis appearance and radiotherapy after surgery. Case 3: An 80-year-old male had lower limb paralysis due to metastasis of Th3 five years and eight months after total gastrectomy for gastric cancer. Paralysis had improved from B of a Frankel classification to C with direct decompressive surgical resection seven hours after the paralysis appearance and radiotherapy after surgery. When the spinal paralysis due to spine metastasis of gastric cancer appeared, the orthopedic surgery for QOL maintenance was considered to be one of the useful medical treatment options.

Trierixin, a new member of the triene-ansamycin group, has been isolated from the fermentation broth of Streptomyces sp. AC654 as an inhibitor of ER stress-induced XBP1 activation. The structure of trierixin was determined on the basis of its spectroscopical and chemical properties. Trierixin possessed a 21-membered macrocyclic lactam, which contains a methylthio-benzenediol structure, and a cyclohexanecarbonylalanine moiety. Trierixin is thus elucidated as 21-thiomethylmycotrienin II.

Signals and organic matrix proteins secreted from the mantle are critical for the development of shells in molluscs. Nacrein, which is composed of a carbonic anhydrase domain and a Gly-X-Asn repeat domain, is one of the organic matrix proteins that accumulates in shells. In situ hybridization revealed that nacrein was expressed in the outer epithelial cells of the mantle of the pearl oyster Pinctada fucata. The recombinant nacrein protein inhibited the precipitation of calcium carbonate from a saturated solution containing CaCl2 and NaHCO3, indicating that it can act as a negative regulator for calcification in the shells of molluscs. Because deletion of the Gly-X-Asn repeat domain of nacrein had a significant effect on the ability of nacrein to inhibit the precipitation of calcium carbonate, it is conceivable that the repeat domain has a primary role in the inhibitory function of nacrein in shell formation. Together these studies suggest that nacrein functions as a negative regulator in calcification in the extrapallial space between the shell and the mantle by inhibiting the precipitation of CaCO3.

The complex thermodynamics that govern noncovalent protein-ligand interactions are still not fully understood, despite the exponential increase in experimental structural data available from X-ray crystallography and NMR spectroscopy. The eukaryotic 20S proteasome offers an ideal system for such studies as it contains in duplicate three proteolytically active sites with different substrate specificities. The natural product TMC-95A inhibits these proteolytic centers noncovalently with distinct affinities. X-ray crystallographic analysis of the complexes of the yeast proteasome core particle with this natural inhibitor and two synthetic analogues clearly revealed highly homologous hydrogen-bonding networks involving mainly the peptide backbone despite the strongly differentiated binding affinities to the three active sites of the 20S proteasome. The natural product and the two analogues are constrained in a rigid beta-type extended conformation by the endocyclic biaryl clamp, which preorganizes the peptide backbone for optimal adaptation of the ligands to the active site clefts and thus favors the binding processes entropically. However, the biaryl clamp also dictates the orientation of the P1 and P3 residues and their mode of interaction with the protein binding subsites. This limitation is optimally solved in TMC-95A with the conformationally restricted (Z)-prop-1-enyl group acting as P1 residue, at least for the chymotrypsin-like active site; however, it critically affects the inhibitory potencies of the analogues, thus suggesting the use of less-rigid endocyclic clamps in the design of proteasome inhibitors that allow for a better presentation of residues interacting with the active site clefts of the enzyme.

New inhibitors of IL-4 signal transduction, designated as TMC-256A1 and C1, were discovered together with TMC-256B1, a previously known dihydronaphthopyrone, from the fermentation broth of Aspergillus niger var niger TC 1629 by using an IL-4 driven reporter gene assay. Based on spectroscopic analyses, TMC-256A1 and C1 were found to be new members of the naphthopyrone antibiotics. TMC-256A1, B1 and C1 inhibited the IL-4 driven luciferase activity with IC50 values of 25 microM, 30 microM and 1.7 microM, respectively in this assay system. Furthermore, these compounds inhibited the expression of germline C epsilon mRNA with IC50 values of 6.6 microM , 34 microM and 0.31 microM, respectively.

The 20 S proteasome core particle (CP), a multicatalytic protease, is involved in a variety of biologically important processes, including immune response, cell-cycle control, metabolic adaptation, stress response and cell differentiation. Therefore, selective inhibition of the CP will be one possible way to influence these essential pathways. Recently, a new class of specific proteasome inhibitors, TMC-95s, was investigated and we now present a biochemical and crystallographic characterisation of the yeast proteasome core particle in complex with the natural product TMC-95A. This unusual heterocyclic compound specifically blocks the active sites of CPs non-covalently, without modifying the nucleophilic Thr1 residue. The inhibitor is bound to the CP by specific hydrogen bonds with the main-chain atoms of the protein. Analysis of the crystal structure of the complex has revealed which portions of TMC-95s are essential for binding to the proteasome. This will form the basis for the development of synthetic selective proteasome inhibitors as promising candidates for anti-tumoral or anti-inflammatory drugs., (Copyright 2001 Academic Press.)

A new transcriptional up-regulator designated TMC-205 was discovered from the fermentation broth of an unidentified fungal strain TC 1630 by using an SV40 promoter-luciferase reporter assay. Based on spectroscopic analyses, its structure was determined to be (E)-6-(3-methyl-1,3-butadienyl)- H-indole-3-carboxylic acid. Expression of the luciferase activity was activated ca. 2-, 4-, and 6-fold by 1, 10, and 100 microM TMC-205, respectively. TMC-205 activated the transcriptional activity in a manner dependent on the presence of the enhancer element of SV40 in its promoter region.

A new antibiotic designated TMC-69 has been isolated from the fermentation broth of a fungal strain Chrysosporium sp. TC 1068. TMC-69 exhibited moderate in vitro cytotoxic activity. TMC-69-6H, a derivative of TMC-69 prepared by hydrogenation, possessed more potent in vitro cytotoxicity than TMC-69, and exhibited in vivo antitumor activity against murine P388 leukemia and B16 melanoma. TMC-69-6H was found to specifically inhibit Cdc25A and B phosphatases.

Four novel proteasome inhibitors, TMC-95A-D (1-4) have been isolated from the fermentation broth of Apiospora montagnei Sacc. TC 1093, isolated from a soil sample. All of the molecular formulas of 1-4 were established as C(33)H(38)N(6)O(10) by high-resolution FAB-MS. Their planar structures were determined on the basis of extensive analyses of 1D and 2D NMR, and degradation studies. Compounds 1-4 have the same planar structures to each other, and are unique highly modified cyclic peptides containing L-tyrosine, L-aspargine, highly oxidized L-tryptophan, (Z)-1-propenylamine, and 3-methyl-2-oxopentanoic acid units. The absolute configuration at C-11 and C-36 of 1-4 was determined based on chiral TLC and HPLC analyses of their chemical degradation products. The ROESY analysis along with (1)H-(1)H coupling constants clarified the absolute stereochemistry at C-6, -7, -8, and -14 of the cyclic moieties. These studies revealed the relationships of 1-4 to be diastereomers at C-7 and C-36.

In our course of screening for novel proteasome inhibitors, TMC-95A and its diastereomers, TMC-95B to D, were isolated from the fermentation broth of Apiospora montagnei Sacc. TC 1093. TMC-95A inhibited the chymotrypsin-like (ChT-L), trypsin-like (T-L), and peptidylglutamyl-peptide hydrolyzing (PGPH) activities of 20S proteasome with IC50 values of 5.4nM, 200nM, and 60nM, respectively. TMC-95B inhibited these activities to the same extent as TMC-95A, while the inhibitory activities of TMC-95C and D were 20 to 150 times weaker than that of TMC-95A and B. TMC-95A did not inhibit m-calpain, cathepsin L, and trypsin at 30 microM, suggesting its high selectivity for proteasome. Taxonomy of the producing strain is also described.

We investigated the clinicopathological correlations among 49 surgically resected thymic epithelial tumors (TET), which were subclassified according to the six subtypes established by the Marino, Kirchner, and Müller-Hermelink system, which were renamed as follows: spindle cell type (medullary thymoma), mixed spindle and polygonal cell type (mixed medullary and cortical thymoma), small polygonal cell type (predominantly cortical thymoma), large polygonal cell type (cortical thymoma), atypical type (well differentiated thymic carcinoma), and cytologically malignant type (high-grade thymic carcinoma). The related categories were grouped for statistical analysis as follows: group 1, spindle cell type and mixed type; group 2, small polygonal cell type and large polygonal cell type; group 3, atypical type; group 4, cytologically malignant type. The association of each group with the presence of myasthenia gravis, tumor stage, and the length of survival was studied. Myasthenia gravis was significantly present in patients with small polygonal type, large polygonal type, and atypical type tumors (groups 2 and 3) (P = 0.003). The tumors in group 1 showed the lowest tumor stage while those of group 4 had the most advanced tumor stage (P = 0.002). The patients in group 4 had the worst prognosis, followed by those in group 3, 2, and 1, in that order. The differences among these groups were statistically significant (P = 0.0003). From our results, we determined that TET can be separated into an extremely low-grade malignancy group (group 1), a low-grade malignancy group (group 2), an intermediate malignancy group (group 3), and a high-grade malignancy group (group 4).

Four new antibiotics, TMC-171A (2), B (3), C (4) and TMC-154 (5) have been isolated from the fermentation of fungal strains Gliocladium sp. TC 1304 and TC 1282, respectively. Spectroscopic and degradation studies have shown that TMC-171s and TMC-154 were new members of the TMC-151 class of antibiotics, unique polyketides modified with a D-mannose and a D-mannitol or a D-arabitol. These compounds showed moderate cytotoxicity to various tumor cell lines.

TMC-86A, B and TMC-96, new 20S proteasome inhibitors with an epoxy-beta-aminoketone moiety, were isolated from the fermentation broth of Streptomyces sp. TC 1084 and Saccharothrix sp. TC 1094, respectively. TMC-86A, B and TMC-96 inhibited the chymotrypsin-like and peptidylglutamyl-peptide hydrolyzing activities of 20S proteasome with the following IC50 values: TMC-86A, 5.1 microM and 3.7microM; TMC-86B, 1.1 microM and 31 microM; TMC-96, 2.9 microM and 3.5 microM, respectively. TMC-86A, B and TMC-96 exhibited the weak inhibitory activity against the trypsin-like activity of 20S proteasome with IC50 values of 51 microM, 250 microM, and 36 microM, respectively. They did not inhibit m-calpain, cathepsin L, and trypsin at 100 microM, suggesting their high specificity for proteasome. Taxonomy of the producing strains is also described.

Four new antitumor antibiotics, TMC-1 A, B, C and D were isolated from a fermentation broth of Streptomyces sp. A-230. Spectroscopic studies have shown that TMC-1 A to D were new members of the manumycin class of antibiotics. These antibiotics showed cytotoxic activities against various tumor cell lines in vitro.

The polyether-macrolide antibiotic, boromycin, was isolated as a potent anti-human immunodeficiency virus (HIV) antibiotic from a fermentation broth of Streptomyces sp. A-3376. Boromycin was found to strongly inhibit the replication of the clinically isolated HIV-1 strain as well as the cultured strain in in vitro laboratory experiments. The mechanism for the anti-HIV activity of boromycin is suggested to involve blocking the later stage of HIV infection, and probably the maturity step for replication of the HIV molecule.

We report a patient with graft versus host disease (GVHD) with mixed chimerism (MC). The patient had chronic myelogenous leukemia and received bone marrow transplantation (BMT) from his elder sister. Eighty days after BMT, erythematous lesions appeared on his chest. Histological examination from the skin lesion revealed lymphocytic infiltration into the upper dermis. Eosinophilic necrotic keratinocytes were scattered through the epidermis. Liquefaction degeneration was also recognized. Sicca syndrome appeared from 110 days after BMT. Detection of host origin Y-chromosome-specific DNA by polymerase chain reaction (PCR) method in bone marrow and peripheral blood showed that all bone marrow samples obtained 6 months from BMT were positive for Y-specific DNA, while peripheral blood became positive in the 60th month after BMT. The host origin normal karyotype (46,XY) in the bone marrow samples was identified for the first time in the 60th month after BMT. These results indicate that host-origin hematopoietic cells survived after BMT.

Patients with atopic dermatitis (AD) were treated with diode low reactive level laser therapy (LLLT), and the following results were obtained. 1) Itchy sensation decreased in 79 of 112 cases (71%) after this therapy. 2) Skin eruptions improved in 69 of 112 cases (62%). 3) There were no side effects during and after LLLT. 4) Major histocompatibility complex (MHC) class II antigen and inter-cellular adhesion molecule (ICAM)-1 expression on epidermal cells decreased after the therapy. 5) The number of CD1 positive epidermal dendritic cells did not significantly change before and after LLLT.

Hyoscyamine 6 beta-hydroxylase (H6H; EC 1.14.11.11) catalyzes the first reaction in the biosynthetic pathway from hyoscyamine to scopolamine in several solanaceous plants. Four monoclonal antibodies were raised against H6H purified from cultured roots of Hyoscyamus niger. The IgG1 antibody mAb5 inhibited H6H activities present in cell-free extracts of H. niger roots and specifically recognized 38-40-kDa proteins from six different scopolamine-producing plant species in Western blot analysis after sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The other three monoclonal antibodies all recognized SDS-denatured H6H protein from Hyoscyamus species, but did not bind to native H6H. Western blot analysis of protein extracts from various tissues of H. niger using these antibodies showed that H6H is abundant in cultured roots, present in plant roots, but absent in leaf, stem, calyx, cultured cells, and cultured shoots. Immunohistochemical studies using monoclonal antibody and immunogold-silver enhancement detected H6H only in the pericycle cells of the young root in several scopolamine-producing plants. Mature roots that underwent secondary growth and lacked the pericycle did not react with the antibody. This pericycle-specific localization of scopolamine biosynthesis provides an anatomical explanation for the tissue-specific biosynthesis of tropane alkaloids and may be important for translocation of tropane alkaloids from the root to the aerial parts.

A 41-year-old man was admitted to our hospital because of hemoptysis and dyspnea. Physical examination disclosed oral ulcer, acne and scrotal ulcer. Although ocular lesions were absent, the needle reaction was positive. Then we diagnosed this case as incomplete type of Behçet's disease. Chest X-ray showed diffuse retinolinear shadows in the entire lung field and an infiltrative shadow in the left middle lung field. CT-scan revealed interstitial shadows in the entire lung field, a wedge-shaped shadow in left S6 and aneurysmic dilatation of the left lower lobe pulmonary artery. Pulmonary angiography showed obstruction and aneurysmic dilatation of the left lower lobe pulmonary artery. Pathological examination of the TBLB specimen revealed thickening of the vessel wall, perivascular infiltrations of mononuclear cells, hemorrhage and slight thickening of alveolar septae. These findings were compatible with pulmonary lesions of Behçet's disease. After administration of corticosteroid, the symptoms and the chest roentgenographic findings improved. Pulmonary lesions of Behçet's disease were discussed.

The relationships both between cholinergic neurons and substance P (SP) and between cholinergic neurons and calcitonin gene-related peptide (CGRP) terminals were examined in the rat sacral intermediolateral nucleus at the light and electron microscopic levels by means of double-immunostaining methods. Cholinergic neurons were labeled by a monoclonal antibody to choline acetyltransferase (CAT) with the avidin-biotin technique and stained bluish-green by indolyl-beta-galactoside reaction products with beta-galactosidase as a marker. On the same sections, SP or CGRP fibers were labeled by polyclonal antisera to SP or CGRP after application of the peroxidase-antiperoxidase (PAP) method and stained brown by the p-dimethylaminoazobenzene (DAB) reaction. After embedding in Epon, light and electron microscopic sections were examined. At the light microscopic level, CGRP-like immunoreactive (CGRP-I) fibers and SP-like immunoreactive (SP-I) fibers were found to pass through the lateral edge of the dorsal horn and then into the dorsal region of the sacral intermediolateral nucleus. In addition, SP-I fibers also extend from the dorsolateral funiculus into the entire sacral intermediolateral region. At the electron microscopic level, many axosomatic and axodendritic synapses were found between CAT-I structures and SP-I terminals in the intermediolateral nucleus, whereas most of the CGRP-I terminals in this area made axodendritic synapses with CAT-I dendrites. These results indicate that cholinergic neurons in the sacral intermediolateral nucleus receive direct synaptic input from SP-I and CGRP-I terminals.(ABSTRACT TRUNCATED AT 250 WORDS)

Re-examination of the strio-nigral substance P (SP) tract by means of an experimental immunohistochemical method in the rats demonstrated the presence of two distinct pathways from the nucleus caudatus putamen (CP) to the substantia nigra (SN). Destruction of the posterior portion of the CP resulted in the disappearance of SP-positive fibers in the SN pars lateralis but not in the SN pars compacta or reticulata. On the other hand, destruction of the ventrolateral portion of the anterior portion of the CP caused the disappearance of SP-positive fibers in the SN pars compacta and pars reticulata but not in the SN pars lateralis. In addition, destruction of the dorsal portion of the anterior portion of the CP, where 3-6 cell islets of SP-positive cells are located, failed to decrease SP-positive fibers in any of the subdivisions of the SN. These findings strongly suggest that SP-positive neurons in the posterior portion of the CP project to the SN pars lateralis (posterior strio-nigral SP tract), SP-positive cells in the lateroventral part of the anterior portion of the CP extend to the SN pars compacta and pars reticulata, but SP-positive cells in the dorsal part of the anterior portion of the CP do not innervate the SN.

Relationships between adrenergic fibers and cholinergic neurons were examined in the rat sacral intermediolateral nucleus using a double-immunostaining method at light and electron microscopic levels. Adrenergic fibers were immunohistochemically stained brown by peroxidase reaction, and cholinergic neurons in the same sections were stained bluish green by beta-galactosidase reaction. In the light microscope, many cholinergic neurons were seen in the intermediolateral nucleus and some of them were surrounded by adrenergic fibers. In the electron microscope, adrenergic fibers made synapses with the cholinergic neurons in the intermediolateral nucleus. These results suggest that adrenergic fibers directly influence the activity of the sacral parasympathetic preganglionic neurons.

The relation between sex hormone levels in blood and ultrastructural changes of ovarian follicles was examined in persistent-estrous rats exposed to continuous illumination (LL) for 100 days. Plasma LH showed a tonic level secretory pattern, and circulating estradiol and estrone concentrations were relatively high, while both levels of FSH and progesterone were low. Various stages of growing and degenerating follicles were observed in the ovary of the LL-exposed rats. The early stage of antral follicle did not seem to possess the ability of steroidogenesis. Theca cells around mature antral follicles contained prominent Golgi apparatuses, plenty of smooth endoplasmic reticulum (ER), abundant free ribosomes and many round-mitochondria. A few newly formed lipid droplets were seen in some of theca cells. Granulosa cells contained much distended rough ER, well-developed mitochondria, several lipid droplets and microfilaments. The theca cells of abnormal follicles with hyperplastic and infolded layers of granulosa cells contained many lipid droplets. However, the development of the smooth ER became hindered with increasing lipid droplets in the theca cell. On the other hand, granulosa cells of abnormal follicles contained greater numbers of lipid droplets than those of antral mature follicles, and were equipped with well-developed cytoplasmic organelles as were those of mature antral follicles. Theca interna cells of abnormal follicles may be more involved in the secretion of androgen, which has already been accumulated in the lipid droplets, than the cells involved in the active synthesis of the hormone, while the granulosa cells may convert its androgen to estrogen. The present findings suggest that both follicles of mature and abnormal types in the LL-exposed rat retain enough capacity of estradiol production and participate in the continued elevation of circulating estradiol, probably resulting in the stimulation of the theca cells by the tonic level of LH and of the granulosa cells by the levels of FSH, which are lower than the basal values during the normal 4-day estrous cycle.

Hyoscyamine is epoxidized to scopolamine via 6beta-hydroxyhyoscyamine in several solanaceous plants. 6,7-Dehydrohyoscyamine has been proposed to be an intermediate in the conversion of 6beta-hydroxyhyoscyamine to scopolamine on the basis of the observation that this unsaturated alkaloid is converted to scopolamine when fed to a Datura scion. To determine whether a dehydration step is involved in scopolamine biosynthesis, [6-(18)O]6beta-hydroxyhyoscyamine was prepared from l-hyoscyamine and (18)O(2) using hyoscyamine 6beta-hydroxylase obtained from root cultures of Hyoscyamus niger L. When [6-(18)O]6beta-hydroxyhyoscyamine was fed to shoot cultures of Duboisia myoporoides R. B(R)., the labeled alkaloid was converted to scopolamine which retained (18)O in the epoxide oxygen. It is concluded that 6beta-hydroxyhyoscyamine is converted in vivo to scopolamine without a dehydration step.