A novel technique for detecting single nucleotide polymorphisms by analyzing consumed allele-specific primers

We present a simple and rapid polymerase chain reaction (PCR)-based technique, termed consumed allele-specific primer analysis (CASPA), as a new strategy for single nucleotide polymorphism (SNP) analysis. The method involves the use of labeled allele-specific primers, differing in length, with several noncomplementary nucleotides added in the 5'-terminal region. After PCR amplification, the amounts of the remaining primers not incorporated into the PCR products are determined. Thus, nucleotide substitutions are identified by measuring the consumption of primers. In this study, the CASPA method was successfully applied to ABO genotyping. In the present method, the allele-specific primer only anneals with the target polymorphic site on the DNA, so it is not necessary to analyze the PCR products. Therefore, this method is only little affected by modification of the PCR products. The CASPA method is expected to be a useful tool for typing of SNPs.     

Isolation of iso-grayanotoxin II from leaves of Leucothoe grayana Max. Its X-ray crystallographic analysis and acute toxicity in mice

The structure of iso-grayanotoxin II, a new diterpenoid from Leucothoe grayana MAX., has been determined as 3 beta,5 beta,6 beta,14 beta,16 alpha-pentahydroxygrayanotox-9(10)-ene by spectroscopic and X-ray crystallographic analysis. The lethal dosage level of iso-grayanotoxin II in mice was lower than that of grayanotoxin III.     

Crystal structures and Mössbauer spectroscopic studies of oxo-centered trinuclear chloroacetate complexes

The crystal structures of oxo-centered trineclear cobalt-iron chloroacetate complex [Co^IIFe ₂ ^III O(CH₂ClCO₂)₆(H₂O)₃]·3H₂O (1) was compared with that of previously reported trinuclear iron complex [Fe^IIFe ₂ ^III O(CH₂ClCO₂)₆(H₂O)₃]·3H₂O (2) which has an isomorphous structure to 1. Compound 1 crystallizes in space group P2₁/n with Z=4 in a unit cell of a=14.826 (4) Å, b=4.536 (8) Å, c=14.000 (4) Å, =100.32 (2)⁰ and V=2968 (11) ų. The structure was refined to R=0.75 and R_w=0.82. The coordination geometries of the three iron atoms are observed equivalent in 1 indicating a static disorder of the position among cobalt and iron atoms. Two distinct Fe^III doublets observed in Mössbauer spectra of 1 become an indistinguishable broad doublet by dehydration of crystal water. On the other hand, no significant line-broadening is observed after the dehydration in complex 2. The results indicate that the dehydration in 2 induces a local environmental change reordering of an electronic configuration around iron atoms, whereas the remaining disordering is reflected in Mössbauer spectrum after the dehydration in 1.     

The economical parameter survey program for long-term transient temperature distribution and stress analysis around the sodium level of the liquid metal fast breeder reactor

The thermal stress around the sodium level of the LMFBR reactor vessel, caused by the axial temperature distribution during heat-up and cool-down transient conditions, is one of the most important problems for a reactor vessel designer, especially in a conceptual design stage. Key parameters relating to plant operating transient conditions and reactor vessel main configuration should be settled carefully, thermal stresses not being allowed to exceed design stress limits. In order to examine the sodium level thermal stress easily and economically, a simple computer program for parameter survey has been developed. This program uses a one-dimensional Fourier series solution in transient temperature distribution analysis, and an analytical stress solution based on shell theory in stress value estimation. This paper presents a simplified and economical calculation method for the axial temperature distribution and stress value in the LMFBR operating transient conditions, and analysis examples obtained with this computer program.

---

Zusammenfassung Der durch die axiale Temperaturverteilung während der Übergangsphase von Aufheizen und Abkühlen verursachte thermische Streß rund um den Natriumfüllstand des LMFBR-Reaktorkessels ist für den Reaktorkesselkonstrukteur, besonders im Entwurfstadiuni eines der wichtigsten Probleme. Die mit den transienten Operationsbedingungen der Anlage und der Gestaltung des Reaktorkessels in Beziehung stehenden Grundparameter müssen sorgfältig ermittelt werden, um zu verhindern, daß der thermische Streß die geplanten Streßgrenzen nicht überschreitet. Um den thermischen Streß um den Natriumfüllstand, leicht und ökonomisch zu bestimmen, wurde ein Komputerprogramm für die Parameterübersicht aufgestellt. In diesem Programm wird von einer eindimensionalen Fourrierreihe zur Analyse der transienten Temperaturverteilung und einer auf der Schalentheorie von Streßwertbestimmungen basierende analytische Lösung Gebrauch gemacht. Im vorliegenden Artikel werden eine vereinfachte und wirtschaftliche Berechnungsmethode für die axiale Temperaturverteilung und Streßwerte bei transienten Arbeitsbedingungen mitgeteilt und mit diesem Komputerprogramm ausgeführte Analysenbeispiele angegeben.

---

, - () , , , . , , , . , , - , . , , .

---

---

Paper presented at the World Conference on Thermal Analysis, Madeira (Portugal) 1986.     

Reactions of hydropolysilanes with alcohols catalysed by tris(triphenylphosphine)chlororhodium

[RhCl(PPh₃)₃] is a very effective catalyst for the cleavage of Si-Si bonds by alcohols.     

Asymmetric synthesis of 2-substituted-3-aminocarbonyl propionic acid

An asymmetric synthetic route to 2-substituted-3-aminocarbonyl propionic acid, which is the significant component of low-molecular-weight renin inhibitors, is described. The key step of this synthesis is diastereoselective alkylation by using chiral oxazolidinone and benzyl bromoacetate.     

Spacer effect of appended moieties for molecular recognition in doubly-sodium anthranilate modified γ-cyclodextrin

-Cyclodextrin with two sodium anthranilate moieties (1) has been prepared as a sensor for detecting organic compounds including terpenoids and steroids. Compound1 shows a pure monomer fluorescence whose intensity is increased or decreased upon addition of the guest species examined. In this system, the sodium anthranilate moieties act either as a spacer, which enables the cyclodextrin to form a 1:1 host-guest complex by narrowing the -cyclodextrin cavity, or as a hydrophobic cap.1 recognizes steroids with much higher sensitivity than terpenoids, in which the appended moieties act as a hydrophobic cap for terpenoids and a spacer for steroids, respectively.     

A new series of antiallergic agents. I. Synthesis and activity of 11-(2-aminoethyl)thio-6,11-dihydrodibenz[b,e]oxepin derivatives.

A new series of 11-substituted 6,11-dihydrodibenz[b,e]oxepin derivatives was synthesized and evaluated for antiallergic activity. Convenient methods for the preparation of sulfides from alcohols were developed. Structure-activity relationships are described. Compound 7, 11-[2-(dimethylamin)ethyl]thio-6,11-dihydrodibenz[b,e] oxepin-2-carboxylic acid hydrochloride, was the most potent in the rat passive cutaneous anaphylaxis test (ED50 = 0.92 mg/kg p.o.). It had a potent inhibitory effect on anaphylactic bronchoconstriction in guinea pigs (ED50 = 0.029 mg/kg p.o.) and H1 receptor antagonistic effect (Ki = 14 nM) with few central nervous system side effects. Additionally, an antagonistic effect against prostaglandin D2-induced contraction of isolated guinea pig trachea (pA2 = 5.73) was an attractive mechanism of action of the new antiallergic agent. Compound 7 was selected for further evaluation as KW-4994.