An enantio- and stereocontrolled synthesis of (-)-mycestericin E via cinchona alkaloid-catalyzed asymmetric Baylis-Hillman reaction.

A new enantiocontrolled synthesis of a potent immunosuppressant(-)-mycestericin E has been accomplished by using cinchona alkaloid-catalyzed asymmetric Baylis-Hillman reaction of an aldehyde with 1,1,1,3,3,3-hexafluoroisopropyl acrylate and Lewis acid-promoted cyclisation of an epoxytrichloroacetimidate as the key steps.

[Chemoradiation therapy for esophageal carcinoma].

Cancer of the esophagus poses a great challenge to the surgical, medical, and radiation professions. Despite standardization of resection and reconstruction techniques, extended lymph node dissection, and advances in perioperative management, the overall prognosis of patients with esophageal cancer undergoing surgical resection has not improved. Preoperative combination chemoradiation therapy in patients with squamous cell esophageal carcinoma has recently received increasing attention, because preoperative chemoradiation therapy, consisting of the concurrent combination of 5-fluorouracil, cis-platinum and radiation, appears to increase the resection rate, the rate of pathological complete responders, and survival time after esophagectomy in patients with locally advanced tumors. Overall in the trials using preoperative concurrent chemoradiotherapy, major antitumor responses have been reported in 40% to 80% of patients, and, consistently, up to 42.1% pathologic complete responses have been seen at esophagectomy. Therefore, preoperative chemoradiation therapy can be anticipated for better prognosis of the patients with locally advanced esophageal cancer.

Alternatively spliced isoforms of the Na+/Ca2+ exchanger in the guinea pig cochlea.

The cochlea has been suggested to express some Na+/ Ca2+ exchangers (NCX), since efficient acoustic transduction requires cytosolic calcium homeostasis. The present study revealed that several spliced isoforms of NCX are expressed in the guinea pig cochlea. Moreover, to determine their localization in the cochlea, microdissected RT-PCR was performed. The guinea pig cochlea was microdissected into three parts (lateral wall, the organ of Corti and modiolus). The cochlear lateral wall and the organ of Corti expressed only a single isoform of NCX1. On the other hand, five isoforms of NCX1 and four isoforms of NCX3 were detected in the cochlear modiolus. The alternative splicing may provide diverse functions for NCX in the cochlea.

Increase of Cl- secretion induced by Kampo medicine (Japanese herbal medicine), Sai-rei-to, in Mongolian gerbil middle ear epithelium.

Sai-rei-to, a type of Kampo medicine (Japanese herbal medicine), has been shown to be clinically effective in treating patients with otitis media with effusion. The effect of Sai-rei-to on the ion transport of the middle ear surface epithelium cultured from the Mongolian gerbil was investigated by using an Ussing chamber. Application of Sai-rei-to to the mucosal bath but not the serosal bath induced an increase in the short-circuit current (I(SC)) in the basal state. The increase in I(SC) was almost completely inhibited by addition of diphenylamine-2-carboxylic acid but not by amiloride, indicating enhancement of Cl- secretion. On the basis of the lack of changes in the intracellular Ca2+ concentration and a sideness of action, the effect of Sai-rei-to on I(SC) is thought to be a direct and selective activation on the apical Cl- channel.

Expression of voltage-dependent chloride channels in the rat cochlea.

Voltage-dependent chloride (ClC) channels have not yet been identified in the cochlea. In this study, an approach utilizing the reverse transcription-polymerase chain reaction (RT-PCR) was devised to clone the cDNA of ClC channels. PCR was performed using degenerate primers corresponding to two highly conserved regions of the ClC channels. By Southern hybridization and sequencing studies, the sequences corresponding to ClC-2 and ClC-3 were found in the cochlear lateral wall, while ClC-1 was not detected. These results suggest that ClC-2 and ClC-3 might be involved in Cl- transport in the cochlear lateral wall.

The Na-K-Cl cotransporters in the rat cochlea: RT-PCR and partial sequence analysis.

Nonsensory epithelial cells of the mammalian cochlea contribute to the production of the endolymph. The presence as well as the function of the Na-K-Cl cotransporter has been suggested, but not yet been proved. To identify the Na-K-Cl cotransporter isoforms expressed in the cochlea, mRNA was extracted from the cochlear lateral wall. After reverse transcription, resulting cDNA was amplified by polymerase chain reaction (PCR) with primers specific for Na-K-Cl cotransporter isoforms. In the rat cochlear lateral wall, mRNA homologous to the mouse bumetanide-sensitive Na-K-Cl cotransporter (mBSC2) was detected. These results suggest that the basolateral localization of the cotransporter in the marginal cell is involved in the secretory process of the endolymph.

Relapsing polychondritis presented as inner ear involvement.

We report a rare case of relapsing polychondritis with an initial symptom of inner ear involvement. This 53-year-old Japanese man experienced a hearing difficulty, tinnitus in both ears, and dizziness of sudden onset, but lacked auricular chondritis at that time, which is the most frequent finding in relapsing polychondritis. Thus it was difficult to reach a correct diagnosis. Steroid therapy, with oral prednisolone 15 mg daily, was effective. Almost two months after we began the steroid therapy, the patient complained of losing interest in his work and reported a hallucination vision on the TV screen, so the dose of prednisolone was decreased to 10 mg. The hallucinations then disappeared, but the serum level of C-reactive protein increased highly. To reduce the dose of prednisolone, we tried low-dose oral methotrexate. However, we had to discontinue it when the patient experienced severe vomiting and diarrhoea. As adjuvant therapy, we then administered Sho-saiko-to, Chinese herbal medicines with few side effects. Symptoms and laboratory abnormalities then improved markedly.

Squamous cell carcinoma of the scrotum.

A patient with psoriasis was found to have a large skin tumor on his scrotum. He had received psoralen and ultraviolet A radiation therapy to control psoriasis. Histopathologic study revealed that the tumor was a well-differentiated squamous cell carcinoma. We present this rare case and suggest that the genitalia be shielded during ultraviolet therapy.

Cloning, characterization, and tissue distribution of porcine SPAI, a protein with a transglutaminase substrate domain and the WAP motif.

The primary and gene structures and tissue distribution of porcine SPAI-2, a protein that belongs to the WAP protein superfamily and has a sodium-potassium ATPase inhibitory activity, were determined by molecular cloning and Northern analysis. A full-length cDNA clone was isolated from a porcine duodenum cDNA library. The cDNA insert encoded a polypeptide of 187 amino acids, which is composed of three domains: a hydrophobic presequence of 21 amino acids, a prosegment of 105 amino acids ending with Asp126, and the mature SPAI-2 sequence of 61 amino acids beginning with Pro127. The prosegment contained 16 repeats of a hexapeptide that is highly homologous to the repetitive sequence found in the transglutaminase domain of the human elafin, an elastase-specific inhibitor that also belongs to the WAP superfamily. The repetitive sequence was demonstrated to be a good substrate of transglutaminase using a recombinant preparation produced in Escherichia coli. A porcine genomic library was then screened for the SPAI gene. Characterization and sequencing of positive clones indicated that the gene is similar to the elafin gene, having 3 exons encoding the 5'-untranslated region and signal sequence, proSPAI, and 3'-untranslated region, respectively. Northern blot analysis revealed intestine-specific expression of SPAI mRNA; the message was especially abundant in the small intestine. ProSPAI was also found in the circulation. The similarity of proSPAI to elafin in the domain structure, the acid-labile nature of the cleavage site (Asp126-Pro127), and the fact that the major form of SPAI in the plasma is proSPAI strongly suggest that proSPAI is not the precursor but rather it is the native form of SPAI. Like elafin, therefore, SPAI appears to be a new type of biologically active substance with a transglutaminase substrate domain that acts as an anchoring sequence.

Effects of hyperthermia on intracellular calcium concentration and responses of cancerous mammary cells in culture.

Effects of hyperthermia on the intracellular calcium concentration (Cai) of an established mouse breast cancer cell line, MMT060562, were studied using fura-2 fluorescence microscopy and the whole-cell clamp technique. A sudden change of temperature from 37 to 45 degrees C induced a transient increase in the fluorescence ratio permeability of the cell membrane and inward current. Deletion of extracellular calcium abolished the fluorescence ratio response to the rise in temperature. Cai of some cells increased after hyperthermia treatment at 44-48 degrees C for 20 min, but the average increase of Cai was negligible. After hyperthermia treatment, spontaneous oscillation of Cai, chemical responses to ATP and bradykinin and the mechanically-induced spreading response diminished. However, the mechanically induced increase of Cai within the stimulated cell remained even after hyperthermia treatment. Suppression of the ATP-induced Cai response recovered to about half the original level within 12 h. Blockage of protein synthesis with cycloheximide (100 microM) had no effect on the recovery. The D-myo-inositol 1,4,5-triphosphate (IP3)-dependent increase of Cai remained intact even after hyperthermia treatment. It is concluded that hyperthermia treatment increases both the permeability of the cell membrane and Cai, but decreases the sensitivity of cells to ATP and bradykinin, presumably due to modification of the signal transduction mechanism.

Control of specific absorption rate distribution using capacitive electrodes and inductive aperture-type applicators: implications for radiofrequency hyperthermia.

A method of controlling the specific absorption rate (SAR) distribution in radiofrequency hyperthermia is proposed. The superposition of the current density associated with capacitively coupled electrodes and that associated with H-field coupled inductive aperture-type applicators modifies the actual current distribution in the heating material. Using a two-dimensional finite element method, we have shown that "focusing" is possible such that the SAR at the center of a phantom can be adjusted to be approximately twice that in superficial regions, even though the wavelength is considerably greater than the dimensions of the phantom.

Effect of PUVA radiation on anaphylactic histamine release from rat dermal tissues.

We have devised a new in vitro model of type I cutaneous anaphylaxis. Male albino rats were sensitized with DNP-Ascaris. Abdominal skin was shaved, and thin, split-thickness slices of skin were cut with a dermatome. The dermis was excised and cut into 100 mg pieces. The dermal tissue was incubated with antigen in Tyrode's solution for 30 min at 37 degrees C. Antigen-induced histamine release from dermal tissue was measured fluorimetrically. Using this system, we measured histamine release from PUVA-irradiated and non-irradiated dermal tissues. A single PUVA irradiation inhibited type I cutaneous anaphylaxis, but did not affect spontaneous histamine release or total dermal histamine. Our model is considered to be useful for investigation of the mechanism of suppression of type I cutaneous anaphylaxis by PUVA.

Inhibitory effects of kampo medicine on Epstein-Barr virus antigen induction by tumor promoter.

We have investigated the possible interaction of Japanese Kampo medicine; Shosaiko-to with Epstein-Barr virus (EBV) induction by TPA (12-O-tetra-decanoyl-phorbol-13-acetate) in EBV-genome harboring cells. The effect of ether extract of Shosaiko-to was also studied on the induction of EBV by superinfection with EBV from P3HR-1 cells (P3H-EBV). Concomitant treatment of EBV-latently-infected cells (Raji and A2L/AH) with TPA and ether extract resulted in effective inhibition of EBV-antigen induction. However, in the experiments to test the influence of ether extract on EBV induction by superinfecting Raji cells with P3H-EBV, ether extract did not affect the percentage of EBV-antigen-positive cells. These data indicate that Shosaiko-to is effective to inhibit antigen synthesis induced by chemical promoters without affecting EBV-antigen induction after superinfection.

[A case report of sacral chordoma].

Sacrococcygeal chordoma is one of the retrorectal tumors. Relative rarity and anatomical location of this may lead to difficulty in diagnosis and surgical treatment. We report a case of sacrococcygeal chordoma successfully treated by high sacral resection by a posterior approach, in which the co-operative efforts of gastroenterological surgeons and an orthopedic surgeon were employed. A 64-year-old man with a long term continued vague anal pain was referred to our hospital. Digital rectal examination revealed an elastic hard mass presacrally. Plain sagittal radiograph, CT, barium enema showed a retrorectal mass and sacral destruction. Diagnosis was confirmed histologically by the specimen taken by open biopsy. Surgical resection was carried out in prone position with the buttocks elevated. The skin incision was upward arched transverse. The lower sacral vertebrae, including S3, were removed en block with the tumor. Bilateral S3 sacral nerve roots were preserved. Postoperative disturbances of the urination and defecation were not seen. High dose radiation therapy, 80 Gray, was done after surgery. Radiation ulcer of the skin was treated by free skin graft, but radiation proctocolitis was not seen. Now he is free from the disease.

Epstein-Barr virus early antigen induction in nasopharyngeal hybrid cells by Chinese medicinal herbs.

The effects of Chinese medicinal herbal drugs (CMH; Daphne genkwa, Wikstroemia indica, Croton oil) were studied for Epstein-Barr virus (EBV) early antigen (EA) induction in established nasopharyngeal hybrid cells. Both ether and water extracts of CMH were found to have inducing activity. However water extracts of the same herbs were not as strong as that of other extracts. The EA positive-cell rate was from 18.2 to 42.2% in ether extracts and 1.0 to 3.8% in water extracts at 10 microgram/ml of the concentration. N-Butyrate alone showed a 40.2% positive rate and in the both treatment of water extracts, a combination effects was seen in induction of the EBV-EA. This in vitro system for the induction of EBV-EA was thought to be useful to determine what is the causal factors for activation of EBV in vivo.

Prostatic malacoplakia: a case report with a review of 49 cases of malacoplakia of various sites in Japan.

We reported a 62-year-old man with malacoplakia of the prostate, and reviewed 49 cases of malacoplakia hitherto observed in Japan in which the lesions originated from the urogenital tract, except for one gastric case. E. Coli was emphasized as a possible causative agent for malacoplakia especially in the urogenital tract. The possible histiocytic origin of von Hansemann cells was stressed by demonstrating cytoplasmic processes and desmosomes in our prostatic case. An adjuvant use of cholinergic agents and ascorbic acid with chemotherapeutic agents was recommended for treating malacoplakia.

Induction of 8-azaguanine- and ouabain-resistant mutants by cyclophosphamide and 1-(pyridyl-3)-3,3-dimethyltriazene in Chinese hamster cells cultured in diffusion chambers in mice.

A host-mediated assay is described for induction of 8-azaguanine-resistant (azgr) and ouabain-resistant (ouar) mutants in Chinese hamster V79 cells cultured in diffusion chambers (DC) in C3H mice. Injection of the hosts with the indirect mutagen/carcinogen cyclophosphamide (CPP) or 1-(pyridyl-3)-3,3-dimethyltriazene (PyDT) caused a dose-dependent increase in mutation frequency at the loci of azgr and ouar in the V79 target cells. Plating efficiency of V79 cells in DC in mice was decreased depending upon the dose of CPP or PyDT given to the hosts. In addition, the relationship between expression time and mutation frequency was examined and discussed. The data support the use of this system as an effective screening procedure for suspected environmental mutagens or carcinogens, especially those that need to be metabolically activated in vivo.