Copolymerization of 1,6-anhydro-sugar derivatives with cyclic monomers, 1. Tri-O-benzyl ether and tri-O-methyl ether of 1,6-anhydro-ß-D-glucopyranose as sugar monomers

The cationic, ring-opening copolymerization of 1,6-anhydro-2,3,4-tri-O-benzyl-ß-D -glucopyranose with epichlorohydrin, 3,3-bis(chloromethyl)oxetane and 1,3-dioxolane was investigated with phosphorus pentafluoride as catalyst at low temperatures. Besides, copolymerization of 1,6-anhydro-2,3,4-tri-O-methyl-ß-D -glucopyranose with epichlorohydrin was studied. Structure and composition of the copolymers were determined by ¹H and ¹³C NMR spectroscopy, indicating that copolymerization occurred in each combination of monomers. Number-average molecular weights of copolymers were in the range of 1 400 to 22 800. From the specific rotation and ¹³C NMR spectrum of copolymers, it was revealed that the ring-opening copolymerization of the benzylated 1,6-anhydro-glucopyranose with the cyclic monomers occurred in a stereoregular manner to give the C-1 carbon of glucose unit with α-configuration. Debenzylation of a copolymer prepared from 1,6-anhydro-2,3,4-tri-Obenzyl-ß-D -glucopyranose and 1,3-dioxolane gave a copolymer composed of free sugar units in the polymer main chain. Assignment of ¹³C NMR spectra of 2,3,4-tri-O-benzyl-ß-D -glucopyranan and of a copolymer of 1,6-anhydro-2,3,4-tri-O-benzyl-ß-ß-glucopyranose with 1,3-dioxolane was attempted.     

Identification of cryptochrome DASH from vertebrates

A new type of cryptochrome, CRY-DASH, has been recently identified. The CRY-DASH proteins constitute the fifth subfamily of the photolyase/cryptochrome family. CRY-DASHs have been identified from Synechocystis sp. PCC 6803, Vibrio cholerae, and Arabidopsis thaliana. The Synechocystis CRY-DASH was the first cryptochrome identified from bacteria, and its biochemical features and tertiary structure have been extensively investigated. To determine how broadly the subfamily is distributed within living organisms, we searched for new CRY-DASH candidates within several databases. We found five sequences as new CRY-DASH candidates, which are derived from four marine bacteria and Neurospora crassa. We also found many CRY-DASH candidates from the EST databases, which included sequences from fish and amphibians. We cloned and sequenced the cDNAs of the zebrafish and Xenopus laevis candidates, based on the EST sequences. The proteins encoded by the two genes were purified and characterized. Both proteins contained folate and flavin cofactors, and have a weak DNA photolyase activity. A phylogenetic analysis revealed that the seven candidates actually belong to the new type of cryptochrome subfamily. This is the first report of the CRY-DASH members from vertebrates and fungi.     

Anterior urethral recurrence of superficial bladder cancer: its clinical significance

The aim of this study was to reveal the clinical features of anterior urethral recurrence in patients with superficial bladder cancer, and to determine the appropriate treatment. Three hundred and three patients with superficial bladder cancer, who were newly diagnosed and initially treated conservatively in our hospital between 1965 and 1990, were followed for at least 5 years and their clinical outcomes were analyzed. Clinical factors, including anterior urethral recurrence, were evaluated statistically regarding tumor progression. Eight patients (2.6%) had anterior urethral recurrence following superficial bladder cancer. Twenty-four patients (7.9%) had tumor progression and 149 (49.2%) had tumor recurrence. In a multivariate analysis using a logistic model, anterior urethral recurrence was the most important factor, followed by histological grade. Four of 5 patients who were treated for anterior urethral recurrent tumors by transurethral resection showed progression and died of the cancer within one year. Two of the remaining three patients who underwent radical cysto-urethrectomy at the time of anterior urethral recurrence survived. Anterior urethral recurrence following superficial bladder cancer is a predictor for rapid subsequent malignant progression. Once there is anterior urethral recurrence, radical intensive therapy, including radical cysto-urethrectomy, should be carried out immediately.     

Overexpression of the Del1 gene causes dendritic branching in the mouse mesentery

In the present study, we established transgenic mice overexpressing Del1, a ligand of integrins, to examine the effect of overexpression of Del1 on vascular morphogenesis. In the wild-type mouse, mesenteric vessels are shaped like rakes consisting of a long stalk and short branches at the periphery. In contrast, those in transgenic mice showed typical dendritic architecture consisting of a few large primary branches with smaller spreading branches. The phenotype of mice overexpressing Del1 suggests the existence of a tissue-specific mechanism for branching morphogenesis in the mesentery.     

Escherichia coli infection induces only fetal thymus-derived gamma delta T cells at the infected site

Intraperitoneal infection of mice with Escherichia coli induced activated TCR gamma delta T cells in the peritoneal cavity. We provide evidence that the E. coli-induced gamma delta T cells are derived only from the fetal thymus on the following grounds. The gamma delta T cells were not induced in athymic nude mice and irradiated bone marrow-transferred mice which lack fetal thymus-derived T cells. However, E. coli infection of fetal thymus-grafted nude mice did induce fetal thymus-derived gamma delta T cells. These results suggest that the fetal thymus-derived gamma delta T cells colonize the periphery during early ontogeny, and are maintained until adult age. The E. coli-induced gamma delta T cells express only the Vdelta1 gene. Vgamma6 was predominantly expressed whereas anti-Vgamma1 and anti-Vgamma4 monoclonal antibodies stained less than 3 % of the cells. Direct sequencing of PCR products revealed that Vgamma6 and Vdelta1 genes expressed by the E. coli-induced gamma delta T cells were invariant sequences identical to those expressed in the fetal thymus. The antigen (Ag) specificity of a T cell hybridoma expressing the fetal type Vgamma6 / Vdelta1(+) TCR could not be identified as the cells failed to respond to lipopolysaccharide, E. coli Ag, mycobacterial heat shock protein 65, or isopentenyl pyrophosphate. These results suggest that the Vgamma6 / Vdelta1(+) gamma delta T cells derived from fetal thymus can participate in immune responses against bacterial infection through recognition of a novel class of Ag which is not yet identified.     

Erythropoietin concentrations are elevated in the peritoneal fluid of women with endometriosis

Erythropoietin (Epo) is an important regulator of erythropoiesis and stimulates the proliferation of early erythroid precursors as well as the differentiation of late erythroid precursors of the erythroid lineage. However, recent studies have indicated that Epo also has angiogenic properties and plays an important role in the oestrogen-dependent cyclical angiogenesis within the mouse uterus. It was therefore postulated that Epo may be an important angiogenic factor in endometriosis. In order to address this hypothesis the concentration of Epo in peritoneal fluid (PF) was determined in patients with or without endometriosis. PF was collected from patients with endometriosis (n = 42) or without endometriosis (n = 18). Detectable concentrations of Epo were found in all PF samples analysed. The concentration of Epo in PF from patients with endometriosis was significantly higher than that in the control group (13.1 +/- 1.2 mIU/ml versus 7.2 +/- 0.7 mIU/ml, mean +/- SE respectively, P < 0.01). Furthermore, in patients with endometriosis the Epo concentrations in PF from patients with stage I disease (n = 17, 16.6 +/- 3.0 mIU/ml) were significantly higher than those with stage II (n = 8, 10.7 +/- 1.2 mIU/ml, P < 0.03), III (n = 13, 8.4 +/- 1.0 mIU/ml, P < 0.01), IV disease (n = 7, 7.5 +/- 1.0 mIU/ml, P < 0.01). These data suggest that Epo may play a role in the pathogenesis of endometriosis particularly in the initiation of the disease.     

Development of analysis of drugs and toxic substances that can play an immediate role in emergency medical service: what is to be analyzed?

Our laboratory was capable of analyzing less than 20 drugs and toxic substances at the time of the establishment of the Center in 1994. Since the poisoning crimes in 1998, such as the curry poisoning with arsenic in Wakayama, the sodium azide poisoning in Niigata, and the potassium cyanide poisoning in Nagano, we have introduced methods for rapid qualitative analysis of arsenic compounds, cyanides and azides, and developed methods for qualitative analysis of three types of surfactants (cationic, anionic, and nonionic) on the basis of the statistics for intoxication patients transferred to the Center. In 1999, the Analysis Method Investigation Committee of the Japanese Society for Clinical Toxicology requested individual medical institutions to analyze 15 selected intoxicating substances, focusing on the following three aspects. 1. Intoxication with a high degree of fatality. 2. Intoxication where analysis plays an immediate role in treatment. 3. Intoxication with a high frequency of requests by clinical physicians for analysis. The selected substances included methanol, barbital drugs, benzodiazepines, tricyclic or tetracyclic antidepressants, methamphetamine, acetaminophen, salicylic acid, bromovalerylurea, organophosphorus pesticides, carbamate pesticides, paraquat, glufosinate, cyanides, arsenic, and theophylline. Responding to the Committee's request, out laboratory has been making efforts so that analysis of drugs and intoxicating substances can play an immediate role in emergency medical service, giving the highest priority to the aforementioned 15 substances. As a result, anyone of us can now rapidly analyze about 35 substances, including those listed by the Society, day and night.     

General Pathology Of Kawasaki Disease

Systemic pathological alterations were studied in thirty-seven autopsied patients with Kawasaki disease. Systemic vasculitis was the most characteristic pathological finding and was present in all the patients. In addition to the vasculitis, there was a high incidence of inflammatory lesions in various organs and tissues: in the heart, endocarditis, myocarditis, and pericarditis; in the digestive system, stomatitis, sialoduct-adenitis, catarrhal enteritis, hepatitis, cholangitis, pancreatitis, and pancreas ductitis; in the respiratory system, bronchitis and segmental interstitial pneumonia; in the urinary system, focal interstitial nephritis, cystitis, and prostatitis; in the nervous system, aseptic leptomeningitis, choriomeningitis, ganglionitis, and neuritis; in the hematopoietic system, lymphadenitis, splenitis, and thymitis. Dermatitis, panniculitis or myositis were also observed in some patients. Therefore, Kawasaki disease is a systemic inflammatory disease which mainly affects the cardiovascular system. These systemic inflammatory lesions are considered to correspond to the variegated clinical manifestaitions. The relationship between Kawasaki disease and infantile polyarteritis nodosa (IPN) were discussed, based on the clinicopathological characteristics.