Causal involvement of mammalian-type cryptochrome in the circadian cuticle deposition rhythm in the bean bug Riptortus pedestris.

Mammalian-type CRYPTOCHROME (CRY-m) is considered to be a core repressive component of the circadian clock in various insect species. However, this role is based only on the molecular function of CRY-m in cultured cells and it therefore remains unknown whether CRY-m is indispensable for governing physiological rhythms at the organismal level. In the present study, we show that RNA interference (RNAi) targeting of cry-m in the bean bug Riptortus pedestris disrupts the circadian clock governing the cuticle deposition rhythm and results in the generation of a single cuticle layer. Furthermore, period expression was induced in cry-m RNAi insects. These results verified that CRY-m functions as a negative regulator in the circadian clock that generates physiological rhythm at the organismal level.

A simple, heat-sterilizable artificial diet excluding animal-derived ingredients for adult blowfly, Lucilia sericata.

Artificial diets have been developed for Lucilia sericata (Meigen) blowfly larvae; however, diets for adults have not yet been developed. An adult diet that excludes animal tissues and animal-derived ingredients and promotes not only ovarian development, but also oviposition, would aid in basic research and maggot debridement therapy. We have successfully developed artificial diets that exclude animal tissues and animal-derived ingredients for L. sericata adults. The outcomes of the diets were comparable with those of a beef liver diet in terms of oviposition, adult survival and number of offspring.

Novel dual inhibitors of 5-lipoxygenase and thromboxane A2 synthetase: synthesis and structure-activity relationships of 3-pyridylmethyl-substituted 2-amino-6-hydroxybenzothiazole derivatives.

As part of our search for novel antiinflammatory drug candidates, we have designed and synthesized a series of 3-pyridylmethyl-substituted 2-amino-6- hydroxybenzothiazoles. Introduction of a 3-pyridylmethyl group into the 2-amino group (type-A) or the benzene ring (type-B) of 2-amino-6-hydroxybenzothiazoles imparted dual inhibitory activity against the production by glycogen-induced peritoneal cells of rat (in vitro) of leukotriene B4 (LTB4) and thromboxane A2 (TXA2), while not significantly inhibiting that of prostaglandin E2 (PGE2). The observed inhibition of the former two arachidonic acid metabolites was indicated to be the result of a direct action on 5-lipoxygenase and TXA2 synthetase by a cell-free in vitro assay. On the other hand, the inhibitory activities against PGE2 production were for most compounds very weak, indicating that they did not inhibit cyclooxygenase. Structure-activity relationship studies concerning the position of the 3-pyridylmethyl group revealed that type-B compounds generally showed about 10-fold stronger inhibitory activity against TXA2 synthetase than type-A compounds. The position of the 3-pyridylmethyl group played an important role in TXA2 synthetase inhibition. When some of these compounds (8, 13a, 26a (E3040), 26b, 27b, and 28b) were orally administered in the rat TNB/ethanol-induced chronic colitis model (100 mg/kg), the production of both LTB4 and TXB2 in the rat colon was reduced (ex vivo). In addition, one type-B compound, 6-hydroxy-5,7-dimethyl-2-(methylamino)-4-(3-pyridylmethyl)benzothiazole (26a), demonstrated a therapeutic effect at treatments of 100 mg/kg po once daily for 11 days and showed almost comparable activity to sulfasalazine at a dose of 500 mg/kg, the reference drug for inflammatory bowel diseases, in this in vivo model.

Structure-activity relationships of (E)-3-(1,4-benzoquinonyl)-2-[(3-pyridyl)-alkyl]-2-propenoic acid derivatives that inhibit both 5-lipoxygenase and thromboxane A2 synthetase.

As part of our research for the development of novel antiinflammatory drug candidates, we have designed and synthesized a series of (E)-3-(1,4-benzoquinonyl)-2-[(3-pyridyl)alkyl]-2-propenoic acid derivatives as dual inhibitors of 5-lipoxygenase (5-LO) and thromboxane (TX) A2 synthetase. In order to increase the absorption after oral administration, we introduced a carboxylic acid moiety into the 1,4-benzoquinone skeleton, which has 5-LO-inhibitory character. Introduction of a 3-pyridylalkyl group at the double bond of the 1,4-benzoquinonyl propenoic acid moiety afforded good to moderate inhibitory activities against the production of leukotriene (LT) B4 and TXA2 while not significantly inhibiting that of prostaglandin E2 by glycogen-induced peritoneal cells of rat (in vitro). The length of the methylene chain of the 3-pyridylalkyl group influenced the inhibition of LTB4 and TXB2 production. An increase of lipophilicity by introducing a more lipophilic alkoxy group did not markedly increase the inhibitory activity on LTB4 production. The position of alkoxy group on the 1,4-benzoquinone skeleton played an important role in TXA2 synthetase inhibition. Compounds such as 20c (E6700) with an appropriate alkoxy group and proper length of methylene side chain, together with a polar substituent (carboxylic acid), showed good inhibition of both 5-LO and TXA2 synthetase and possess a variety of pharmacologically beneficial effects.

Involvement of tenascin-C in proliferation and migration of laryngeal carcinoma cells.

Tenascin-C (TN-C) is an extracellular matrix glycoprotein upregulated in various pathological processes. In this study, we investigated its distribution in dysplasia and carcinoma of the human larynx using immunohistochemistry and in situ hybridization (ISH) techniques. In all cancer tissues, TN-C immunostaining was markedly increased in the stroma, especially around the cancer cell nests. In addition, cytoplasmic staining of cancer cells was also observed in 62.5% of the invasive cases, the cells being distributed in the periphery of the nests adjacent to the stroma. TN-C mRNA signals in cancer cells were detected in all six cases examined by ISH. Furthermore, in vitro evaluation of the roles of TN-C demonstrated an increase in the proliferating cell fraction in a dose-dependent manner. In a wound closure assay, the addition of TN-C promoted migration. We conclude that TN-C secreted by cancer cells may be involved in their proliferation and migration in an autocrine fashion.

Structural and functional studies on biliverdin-associated cyanoprotein from the bean bug, Riptortus clavatus.

Structural and functional analysis were performed on biliverdin-associated cyanoprotein (CP) from the hemolymph (CP-1 to CP-4) and eggs (CPegg, identical to CP-1) of the bean bug, Riptortus clavatus. Isoelectric focusing analysis of purified CPegg and CP-4 revealed that they are composed of a single alpha subunit and a more acidic beta subunit respectively. N-terminal amino acid sequencing revealed that there are six different amino acid residues between the two subunits up to the 29 cycles determined, indicating that they are encoded by different genes. Appreciable sequence similarities in N-terminal region are found between the CP subunits and several insect hexamers. By chemical cross-linking analysis these CPs were demonstrated to have hexameric structures. Two dimensional peptide mapping revealed that alpha and beta subunits share common structures in part and that CP-2 and CP-3 are hybrid molecules bearing both alpha and beta subunits. From these results the molecular structure of CPs was established as follows: CP-1 (CPegg) = alpha 6; CP-2 = alpha 4 beta 2; CP-3 = alpha 2 beta 4; CP-4 = beta 6. In addition, using 125I-labeled CPegg and CP-4, in vivo incorporation into several tissues was examined during the nymphal-adult development. During the late phase of the 5th instar, CP-4 was sequestered preferentially over CPegg by the fat body, while after emergence both proteins were suggested to be incorporated into the newly-formed cuticle. CPegg was sequestered massively by the developing ovaries whereas no appreciable incorporation of CP-4 was observed.

Trigeminal pain caused by a parapontine epidermal cyst.

A case of trigeminal neuralgia caused by a parapontine epidermal cyst is reported. The trigeminal inlet zone at the pons was completely surrounded by the tumor. No vascular element was detected around the trigeminal nerve root. Neurological symptoms were completely normal except for typical tic douloureux. Pain was controlled initially by carbamazepine, later by radiofrequency trigeminal nerve coagulation, and finally by microsurgical decompression. Review of the literature suggested the importance of lesions of the middle fossa as another etiological factor. When surgery is performed by the posterior route, the possibility of overlooking these lesions may exist.

[Toxicity study of sodium N-[2-[4-(2,2-dimethylpropionyloxy) phenylsulfonylamino] benzoyl] aminoacetate tetrahydrate (ONO-5046.Na) (5). 6-month repeated dose intravenous toxicity study in dogs with 1-month recovery test].

A 6-month repeated dose toxicity study with 1-month recovery test of sodium N-[2-[4-(2,2-dimethylpropionyloxy) phenylsulfonylamino] benzoyl] aminoacetate tetrahydrate (ONO-5046.Na), a novel neutrophil elastase inhibitor, was conducted in beagle dogs. The dogs of both sexes were administered ONO-5046.Na intravenously at a daily dose of 0 (vehicle control), 7.5, 15 or 30 mg/kg. In the 15 mg/kg and above groups, transient ataxic gait was observed. It is considered that this symptom could be attributed to the pharmacological effect of ONO-5046.Na. Macro- and microscopic hemorrhage at the injection site was observed in the ONO-5046.Na treated groups. However, it is considered that these findings could be attributed to the long-term repeated dosing procedure, and were not toxic changes. There were no treatment-related changes in body weight, food consumption, ophthalmology, urinalysis, hematology, blood chemistry, electrocardiography and organ weights. These results indicate that the NOAEL of ONO-5046.Na in dogs is 30 mg/kg/day for both sexes in this study.

The protective effect of cysteine on chemical-induced liver injury in rats.

The possible protective effect of cysteine on chemical-induced liver injury was studied in rats in vivo and in vitro. There was no increase in the activity of serum glutamic oxaloacetic transaminase (GOT) of rats pretreated with cysteine (1.2 g/kg, p,o.) followed by 0.25 ml/kg carbon tetrachloride (CCl4), d-galactosamine (GalN) or alpha-naphthylisothiocyanate (ANIT). However, rats pretreated with cysteine followed by 0.5 ml/kg CCl4 were not protected. The content of cytochrome P-450, activity of aminopyrine N-demethylase or serum ratio of 5,5-dimethyl-2,4-oxazolidinedione (DMO) to trimethadione (TMO) (DMO/TMO ratio) after CCl4, GalN or ANIT were not altered by pretreatment with cysteine. However, pretreatment with cysteine prevented changes in the content of cytochrome P-450, activity of aminopyrine N-demethylase and DMO/TMO ratio in serum as well as the activities of serum GOT and GPT when the rats were treated with bromobenzene (BZ). The degree of lipid peroxidation from CCl4 was markedly reduced by the presence of 10(-4)M cysteine. These results suggest that cysteine has a protective effect on chemical-induced liver injury produced via epoxide metabolites.

[Clinical study of S 6472 granule preparation (sustained-release cefaclor) in chronic respiratory airway infections].

S 6472 granule preparation, a sustained-release preparation of cefaclor, was administered to 21 patients with chronic respiratory airway infections for its clinical study; a daily dosage between 750 and 1,500 mg was orally given in 2 divided doses after breakfast and dinner for a duration of 3 to 14 days. Clinical effects were good in 15 cases, fair in 1 case, poor in 4 cases and unknown in 1 case. No side effects were observed except for a case of impaired appetite. There appeared to be no abnormal laboratory test valued due to the drug.

[Clinical studies of cefpodoxime proxetil in respiratory tract infections].

Twelve patients with respiratory tract infections were treated with cefpodoxime proxetil (CS-807, CPDX-PR), a new cephem antibiotic. It was given orally at a dose of 200 mg 2 times a day for 4 approximately 15 days. Its clinical effects were evaluated as excellent in 1 case, good in 9 cases and poor in 2 cases. The efficacy rate was 83.3%. Its bacteriological effects were evaluated as eradication in 5 strains and decrement in 1 strain. The eradication rate was 83.3%. No adverse reactions and disorder of laboratory findings due to CPDX-PR were observed.

[Multiple myeloma presenting with a solitary cranial tumor with subdural extension: a case report].

We report a case of multiple myeloma presenting with a solitary cranial tumor in the frontal region, extending from subcutaneous tissue to subdural space. To our knowledge, invasion of a tumor beyond the dura mater has never been described in case of multiple myeloma presenting with a solitary calvarial tumor. In the present case, the subdural extension was clearly visualized by several diagnostic means. A 53-year-old female patient visited the clinic of our University Hospital, complaining of a left frontal mass in May 1991. The mass grew rapidly, and she was hospitalized in June. On admission, neurological examination showed nothing abnormal. Laboratory studies showed normocytic normochromic anemia. Protein electrophoresis disclosed hypergammaglobulinemia with S-spike, and serum paraprotein was specific to IgA with lambda light chains by immunoelectrophoresis. Urinary Bence-Jones protein was not detected. An osteolytic lesion visualized in the frontal bone on plain skull radiographs showed destruction of the frontal bone, and an enhanced mass extending from the epidural to subcutaneous space was shown by computed tomography. The mass had compressed the frontal lobe. T1-weighted magnetic resonance images of the area showed isointensity signals, homogeneously enhanced with gadolinium-diethylenetriamide pentaacetic acid. Subdural extension in the deep area of the tumor was suggested. At operation, we confirmed infiltration of the tumor from the dura mater into the subdural space. Postoperative biopsy of bone marrow of the iliac bone demonstrated myeloma cells. The tumor was histologically diagnosed as plasmacytoma. The patient was highly resistant to postoperative combined therapy, and started on a fatal course leading to respiratory insufficiency caused by interstitial pneumonia in April 1992.(ABSTRACT TRUNCATED AT 250 WORDS)

[A case of microsurgically removed cavernous angioma in the midbrain].

The authors presented a case of microsurgically removed large cavernous angioma in the midbrain. A 46-year old woman was admitted to our service with bilateral nuclear oculomotor pareses and mild tetraparesis. MRI revealed a large round well-circumscribed high signal-intensity lesion on both T1 and T2 weighted image with low signal rim in the midbrain. This lesion was diagnosed preoperatively as a cavernous angioma. The operation was performed in three steps with intermittent hemorrhages by interhemispheric transcallosal-hippocampal commissure--velum interpositum-third ventricular approach and subsequent two infratentorial supracerebellar approaches and finally complete removal was performed. Histological examination of the surgical specimen revealed as a cavernous angioma having abnormal vessels with honeycomb appearance. The patient survived, although remains moderately disabled 6 months after the last operation.

[Immunochemotherapy of human gliomas transplanted into nude mice].

In the first part of this paper, various chemotherapies were performed against oligodendrogliomas subcutaneously transplanted in to nude mice. Vincristine (VCR), adriamycin, and 1000 rads irradiation were effective against this tumors. Concerning these two drugs, dose response effect was observed. And the effect of 1 mg/kg injection of VCR roughly corresponded to that of five weekly injections of 0.2 mg/kg. In the second part of this experiment, single or combined effects of VCR and immunotherapeutic agents including OK-432 (OK), PSK, and recombinant leucocytic interferon (IFN) were examined. Two glioma lines including oligodendroglioma and glioblastoma were used. Following results were obtained from this experiment: 1) Effect of OK and VCR against oligodendrogliomas were as follows: control less than OK local injection (Local) less than OK intraperitoneal injection (IP); VCR; OK (IP X 2) less than VCR + OK(IP) less than VCR + OK (IP X 2). Effects of OK and VCR were expressed in order of their effects against glioblastomas: control less than VCR less than OK (IP); OK(IP X 2); OK(Local) less than VCR + OK (IP); VCR + OK (IP X 2). Effects of PSK and VCR against glioblastomas were as follows: control; PSK (Local) less than VCR less than VCR + PSK (Local) less than VCR + PSK (IP). Effects of IFN and VCR against oligodendrogliomas were as follows: control; IFN (IP) less than VCR less than IFN (Local) less than VCR + IFN (IP); VCR + IFN (Local). Effects of IFN and VCR against glioblastomas were as follows: control less than IFN (IP) less than VCR; IFN (Local); VCR + IFN (IP).(ABSTRACT TRUNCATED AT 250 WORDS)

[CO diffusing capacity during continuous negative extra-thoracic pressure].

Eight healthy males were studied to compare CO diffusing capacity (DLCO) during spontaneous breathing with that during continuous negative extra-thoracic pressure (CNETP). Mean DLCO was 33.0 +/- 5.1 ml.min-1.mmHg-1 during spontaneous breathing and 33.5 +/- 4.5 ml.min-1.mmHg-1 during CNETP with an end-expiratory negative extra-thoracic pressure (EENETP) of -20 cmH2O, and there was no significant difference between them (P less than 0.05). Pulmonary capillary blood volume, which was measured only in a male, was 76.7 ml during spontaneous breathing and 80.5 ml during CNETP. This change dose not seem to be significant. The results suggest that the effect of pulmonary diffusing capacity changes during EENETP on improvement of oxygenation may not be significant.

[A case of eosinophilic pneumonia possibly due to ifenprodil].

A 69-year-old woman with a history of subarachnoid hemorrhage was started on ifenprodil for dizziness. Three weeks later, fever, cough, chills, dyspnea and skin eruption developed. A chest radiograph showed bilateral ground-glass shadows. Blood tests showed a white cell count of 14,400/mm3 with 32% eosinophils and a C reactive protein (CRP) level of 20 mg/dl. The arterial blood gases on room air were as follows: pH 7.45, PaCO2 33 torr, and PaO2 56 torr (Table 1). Ifenprodil was withdrawn and intravenous meropenem and minocycline administration was started on admission. Her fever improved rapidly and the CRP decreased, but hypoxemia and hypereosinophilia persisted. On the third hospital day, she underwent bronchoscopy with bronchoalveolar lavage (BAL). The differential count of BAL cells was 63% eosinophils, 15% lymphocytes, 21% macrophages, and 1% neutrophils. Intravenous methylprednisolone 250 mg/day for 3 days was commenced, leading to a clinical improvement. She received oral prednisolone (30 mg/day) for the next 4 days, and was then discharged without any symptoms. She has had no recurrence since. Both the drug lymphocyte stimulation test and the skin test for ifenprodil were negative.

[Drug-induced pneumonitis caused by sulfamethoxazole-trimethoprim].

A 33-year-old man was treated with sulfamethoxazole-trimethoprim (SMT-TMP) for an infection in the cervical vertebrae by methicillin-resistant Staphylococcus aureus (MRSA). Two weeks later a fever of 39 degrees C appeared, and a productive cough, hemosputum, and dyspnea developed a further three weeks later. Chest radiographs showed bilateral ground-glass opacity. Cell differentiation of bronchoalveolar lavage fluid (BALF) revealed increases of lymphocytes and eosinophils, and the CD4/CD8 ratio of the BALF lymphocytes was decreased. A thoracoscopic lung biopsy specimen showed fibroedematous thickening of the alveolar walls, hypertrophic alveolar cells, and cell infiltration with neutrophils and lymphocytes. The pathological diagnosis was nonspecific interstitial pneumonia, group II. The fever resolved 6 days after discontinuance of SMX-TMP. The lymphocyte stimulation test for SMX-TMP gave a positive result. Administration of glucocorticoid improved both the symptoms and the laboratory data.

[A case of pneumonitis possibly due to isocyanate associated with high levels of serum KL-6].

A 47-year-old automobile painter developed dry cough, chills and dyspnea after four months of painting work. The spray contained isocyanates (HDI). A chest radiograph showed bilateral ground-glass shadows. The serum KL-6 level was very high: 11,100 U/ml. Marked lymphocytosis and a striking decrease in the CD4/CD8 ratio were observed in the bronchoalveolar lavage fluid. Transbronchial lung biopsy specimens showed alveolitis and bronchiolitis. Cessation of exposure to isocyanates improved the symptoms and laboratory data. We suspected that the patient was suffering from hypersensitivity pneumonitis induced by isocyanates. Specific IgG antibodies for TDI, MDI, and HDI were not demonstrated by enzyme-linked immunosorbent assay. Because abnormal shadows in the chest radiograph did not improve quickly after admission, we administered glucocorticoids which improved the symptoms, the chest radiograph findings, and the serum KL-6 level.

[Study on postoperative local chemotherapy of malignant brain tumors using ACNU and PSK].

There have been many attempts to treat patients with malignant brain tumors represented by glioblastomas using nitrosourea (NU) derivatives such as BCNU and CCNU but the clinical results are not so remarkable compared with previous reports concerning experimental studies. The reason for an efficacy of NU derivatives in brain tumors is considered to be its higher lipid solubility which makes the drug crossing the BBB easily. On the other hand, there is some evidence that higher lipid solubility did not guarantee NU to reach always to all portions of solid tumor after systemic administration. We have performed a chemotherapy of malignant brain tumors using ACNU for five years. This drug is not only lipid but also water-soluble in some grade; therefore, the drug is administrated intravascularly and locally with ease. Nine cases of malignant gliomas were treated with local chemotherapy employing ACNU and two cases of glioblastomas are surviving now over five years and about four years, respectively. From the anatomical standpoint of view, it is considered to be necessary for local chemotherapy of brain tumors to possess some pathognomonic characters such as cyst formations, central necrosis and localized cortico-meningeal adhesions, which are rather frequently found in malignant gliomas and are suspected also easily by CT examination preoperatively. A local chemotherapy in the present study has been performed using 10 to 50 mg of ACNU through an indwelling catheter inserted during operation. No general toxicity occurred in all patients except one, who experienced purulent meningitis after long-term drainage. In our study on concentration of ACNU, intracarotid injection resulted in higher concentration in brain tumor tissue than intravenous injection, but these concentration considered to be not high enough to suppress the tumor cell growth. On the other hand, the intracavitary concentration of ACNU at 24 hr after drug administration was high enough to suppress the tumor cell growth.