[Time resolved Raman spectroscopy--stopped flow and pulse radiolysis methods].
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Biomedical subjects
Publications and source records attributed to Y Nishimura.
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Solution conformation in different conditions of r(CGCGCG) has been studied by a Raman spectroscopic method. In NaCl solution, r (CGCGCG) takes only an A-form duplex in which guanosine and cytidine have C3'endo-anti conformation even at 5M salt concentration. In much higher ionic strength condition (5M NaCl plus 1M MgCl2 or 6M NaClO4), it undergoes a transition to a left-handed Z-form. The Raman spectrum of the Z-form RNA was found to be very similar to that of Z-form DNA, suggesting that Z-RNA involves a C3'endo-syn guanosine and an in between form of C2'endo-Cl'exo-anti cytidine.
Raman spectra of nine crystals of known structures which involve guanosine moieties with various conformations have been observed. It has been established that a guanosine residue with the C3'endo-anti conformation gives a strong Raman line at 666 +/- 2 cm-1. It has also been found that the residue with 04'endo-anti gives a strong Raman line at 682 cm-1, and C3'exo-syn at 616 cm-1. The usefulness of these structure-spectrum correlations in the conformation studies of polynucleotides are shown.
Electrophysiological studies on the entopedunculo- and cerebello-thalamic projections were performed by intracellular recordings in the thalamic VA, VL and VM nuclei of cats under sodium pentobarbital anesthesia. Identification of the thalamic neurons were performed electrophysiologically by antidromic activation on stimulation of the precruciate cortex (areas 4 and 6) and the caudate nucleus, and morphologically by intracellular staining with HRP through recording microelectrodes. One hundred and sixty-three neurons were collected in the VA, VL and VM nuclei. In 79 neurons penetrated in the medial and ventral parts of the VA and VL nuclei, stimulation of the entopeduncular nucleus induced monosynaptic IPSPs (latency of 1.1-3.5 ms, mean 2.07 ms). Sixteen neurons were identified as thalamo-cortical relay neurons and 3 were activated only orthodromically by precruciate stimulation. Seventy-eight neurons located dorsolaterally to the entopeduncular-influenced neurons received only cerebellar EPSPs. Only 6 neurons showed convergence of entopeduncular and cerebellar inputs. They were scattered around the border between the entopeduncular and cerebellar projection areas. Sixteen neurons could be stained intracellularly by HRP injection. From the pattern of dendritic arborization, two types of neurons can be distinguished: neurons whose dendrites spread radially in all directions and neurons whose dendrites extend mainly along the long axis of the soma for a long distance in the frontal plane, respectively. The former are relay cells to the cerebral cortex or the caudate nucleus (i.e. projection neurons) and the latter appear to be interneurons in the thalamus.
The UV absorption of HMM (heavy meromyosin) was measured at various temperatures with a computerized spectrophotometer system. HMM showed temperature-induced absorption changes in the presence and absence of nucleotides. The temperature-induced absorption change at 293 nm, which is due to conformational changes around the tryptophan residues of HMM, was enhanced in the presence of nucleotides. The temperature-induced difference spectra of HMM + AMPPNP relative to HMM obtained by using a conventional spectrophotometer [(1977) J. Biochem. (Tokyo) 81, 313-320] could be reproduced by subtracting the temperature-induced spectral changes of HMM from those of HMM + AMPPNP.
We have analyzed HR changes greater than 20% among 334 patients and 942 cerebral angiographies. A tachycardial effect was seen in 14.9% of patients, while a bradycardial effect was seen in 7.1% including two patients having cardiac standstill (0.5%). These two patients were examined without atropine premedication after subarachnoid hemorrhage. Patients under 19 years of age, unpremedicated with atropine sulfate and suffering from subarachnoid hemorrhage secondary to ruptured aneurysm or arteriovenous malformation showed a significantly high incidence of bradycardia. On the other hand, patients with the neoplastic disease and having an initial sinus bradycardia showed a significantly high incidence of a tachycardial effect.
Circulating interferon levels increased and persisted for prolonged periods of time when groups of mice were stimulated with a mixture of Newcastle disease virus (NDV) and EDTA, or pretreated with EDTA and then injected with NDV, instead of injection with NDV alone.
The title compounds having nitro, amino, cyano, chloro, or fluoro as the C-6 substituent were prepared. Introduction of the chloro and cyano groups at C-6 was accomplished by the Sandmeyer reaction of 6-amino-1,8-naphthyridine derivatives 9 via their 6-diazonium salts. The reaction was extended to the synthesis of the 6-fluoro analogues, involving the Balz-Schiemann reaction of the diazonium tetrafluoroborate. Furthermore, a series of the 1-ethyl, 1-vinyl, 1-(2-fluoroethyl), and 1-(difluoromethyl) analogues of 7-substituted 6-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acids was prepared. 1-Pyrrolidinyl and, particularly, N-substituted or unsubstituted 1-piperazinyl groups were introduced as the C-7 variants. As a result of this study, 1-ethyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl)-1, 8-naphthyridine-3-carboxylic acid (named enoxacin, originally AT-2266) was found to show the most broad and potent in vitro antibacterial activity, an excellent in vivo efficacy on systemic infections, and a weak acute toxicity. Structure-activity relationships of compounds with variations of substituents at C-1, C-6, and C-7 are also discussed.
The title compounds (28-56) with an amino- and/or hydroxy-substituted cyclic amino group at C-7 were prepared with 1-substituted 7-chloro-, 7-(ethylsulfonyl)-, and 7-(tosyloxy)-6-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine-3- carboxylic acids and their ethyl esters (3-7) with cyclic amines such as 3-aminopyrrolidine. The N-1 substituent includes ethyl, vinyl, and 2-fluoroethyl groups. As a result of in vitro and in vivo antibacterial screenings, three compounds, 1-ethyl- and 1-vinyl-7-(3-amino-1-pyrrolidinyl)-6-fluoro- 1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acids (33a and 33b) and 1-vinyl-7-[3-(methylamino)-1-pyrrolidinyl] analogue 34b, were found to be more active than enoxacin (2) and to be worthy of further biological study. Structure-activity relationships are discussed.
The temperature-dependent precipitability of a monoclonal IgG3 kappa cryoimmunoglobulin (Jir) without known antibody activity is shown to be affected by various physico-chemical factors, such as protein concentration, pH value and NaCl concentration. The molecular properties characterizing this protein (carbohydrate and amino acid compositions, peptide constitutions and susceptibility to enzymatic proteolysis) are described. The cryoprecipitability of the protein was completely lost upon papain hydrolysis, and none of the isolated fragments, Fab-Fc, Fc, and Fab, showed any precipitating activity. In the cryo-coprecipitation assay using the 125I-labeled fragments, it was demonstrated that the association activity with intact Jir protein was still retained on the Fab-Fc and Fc fragments, but not on the Fab fragment. The evidence suggests that a specific interaction may be involved in the primary intermolecular association required to form the cryoprecipitate at temperatures below the critical point, and that one of the pairing sites resides on the Fc portion of the protein molecule.
Myosin head modified with p-chloromercuribenzoate (CMB) forms rigor-like complex with actin in the presence of ATP. Actomyosins with CMB-modified myosin were reconstituted to study the effect of rigor-like complexes on superprecipitation. As native myosin was increasingly replaced by CMB-modified myosin, superprecipitation of the actomyosin was strongly suppressed. Further, the suppression of superprecipitation occurred in a different fashion depending on how CMB-modified myosin was incorporated in myosin filaments of the reconstituted actomyosin. The present results indicate that superprecipitation requires the dissociation of actin and myosin head to take place (i.e., the presence of molecular rearrangements of actomyosin network), and further suggest that superprecipitation is associated with dynamic rearrangements of actomyosin network along myosin filaments.
The cutting modes of restriction endonucleases which recognize tetradeoxyribonucleotide sequences are classified into two groups. d(GGCC) and d(CGCG), for example, are cut to produce blunt ends, while d(CCGG) and d(GCGC) are cut to produce two-base-long cohesive ends. A conformational analysis by the Calladine-Dickerson method indicates that d(GGCC) and d(CGCG) should have a roll angle of successive base-pairs open towards the major groove at the central (second) base-pair step. On the other hand, d(CCGG) and d(GCGC) have such open roll angles at the first and third base-pair steps. It is suggested that, in general, the cutting mode of a tetramer-specific enzyme depends primarily upon the substrate conformation, rather than upon the enzyme. Similar correlations between the mode of action and substrate conformation are also suggested for hexamer-specific enzymes.
The gene frequency (+/- SD) of SB alleles was estimated in the Japanese population as follows: SB1 0.00, SB2 0.195 +/- 0.027, SB3 0.070 +/- 0.017, SB4 0.091 +/- 0.019, SB5 0.335 +/- 0.034 and SB blank 0.308 +/- 0.035. The absence of SB1 allele, decrease of SB4 allele and increase of SB5 allele are characteristic features of the SB system in the Japanese population as compared with Caucasians.
Cytotoxicity inhibition assay was established for the screening of a monoclonal antibody to HLA antigen. The assay involved the inhibition of typing cells with hybridoma culture supernatant and with F(ab')2 fragment of sheep anti-mouse IgG. Using the assay and the conventional microcytotoxicity test, an anti-HLA-A2 monoclonal antibody was screened.
The present study was performed to determine whether serum levels of phenobarbital were representative of their concentrations in the brains obtained from 11 autopsied patients, including premature and full-term babies, infants and children. Estimation of phenobarbital concentrations in plasma and organs was performed by high-pressure liquid chromatography. Brain phenobarbital concentrations correlated well with serum levels (r = 0.82, p less than 0.01, n = 11), giving a regression line with a slope of 0.75. It can be concluded that estimating the serum phenobarbital concentration during clinical pediatric practice gives a good indication of the brain concentration.
The hepatic esterase activities towards chloramphenicol succinate were determined in the tissues from 45 human subjects, including 18 electively aborted fetuses, 5 premature and 8 full-term newborn babies, 8 infants and 6 adults. The enzyme activities in the tissues from the fetuses and neonates were significantly lower compared with those obtained from the infants and adults. This suggests that the activity showed postnatal development. Kinetic studies of the esterase activity revealed that hepatic Km values were similar to those of the lung, but renal Km values were about 3 times higher than those of the liver and lung. In each organ, no age-related changes in Km values were observed. However, all the Vmax values in each organ showed developmental increases. Therefore, the liver plays the most important role in hydrolysis of chloramphenicol succinate when the weight of the organ is taken into consideration.