Hybridization between oxy-peptide nucleic acids and DNAs: dependence of hybrid stabilities on the chain-lengths, types of base pairs, and the chain directions.

Oxy-peptide nucleic acids (OPNAs) of [-NH-CH(CH2-CH2-Base)-CH2-O-CH2-CO-]-type main chain with four different types of nucleobases (Base = A, G, C, and U) or with an abasic side group (X) were synthesized. Melting curves of the 1:1 hybrids of o(A(n))-d(Tn)) pairs with n = 6, 9, 12, and 15 showed very sharp transitions at high Tm values, particularly for long chains, indicating that nearly optimum matching is attained in the structure of the o(A(n))-d(Tn) hybrids. Effect of different types of base pairs on the hybrid stabilities was examined for the o(A4NA4)-d(T4N'T4) 1:1 mixtures where N is A, G, C, U, or X and N' is A, G, C, or T. In all series of the hybrids the complementary pairs showed the highest Tm values. The Tm values of the complementary pairs were about 35 degrees C when purine bases were inserted as the N group in the OPNA, but they were 20-23 degrees C when pyrimidine bases were inserted. The melting curves of the hybrids with a single mismatch were similar to those with a single X-N' pair, suggesting that the mismatch base pairs have been ignored in the hybrids. All complementary OPNA-DNA hybrids showed higher Tm values and sharper transitions than the corresponding DNA-DNA hybrids. The OPNA-DNA hybrids favor a parallel direction i.e., the N-terminal of OPNA is directed to the 5'-terminal of DNA.

Synthesis of oxy-peptide nucleic acids with mixed sequences.

Syntheses of N-Fmoc delta-amino acids with an ether linkage in the main chain and six different nucleobases on the side chain, Fmoc-NH-C*H(CH2-CH2-B)-CH2-O-CH2-COOH (B = N6-benzoyladenine, thymine, uracil, N-benzoylcytosine, guanine, and N2-isobutyrylguanine) are described. The delta-amino acids were prepared through 8-12 step synthesis starting from L-homoserine and could be linked together to form novel peptide nucleic acids (oxy-PNAs = OPNAs) by solid-phase peptide synthesis.

Mechanism of relaxant effect of clonidine in isolated bovine tracheal smooth muscle.

The relaxant effect of clonidine and the possible involvement of imidazoline I1 receptors in bovine tracheal smooth muscle (BTSM) were examined. Clonidine caused concentration-dependent significant relaxation in BTSM precontracted with 0.1 or 1 microM carbachol (CCh) but not in 72.7 mM KCl-induced contraction. The relaxation in CCh-contracted BTSM was inhibited by yohimbine (1 microM) and idazoxan (10 and 30 microM) but not by tetrodotoxin, indomethacin and other adrenoceptor antagonists. Oxymetazoline (0.1-100 microM) and phentolamine (0.1-100 microM) caused concentration-dependent relaxation, which was attenuated by idazoxan (10 microM). Norepinephrine (0.1-100 microM) produced concentration-dependent relaxation, which was completely abolished by propranolol (10 microM) but not by yohimbine (1 microM). In fura-PE3/AM-loaded BTSM, CCh and 72.7 mM KCl increased intracellular calcium concentration ([Ca++]i) followed by contraction. The high K+-induced increase in [Ca++]i was not affected by clonidine. In CCh-stimulated BTSM, clonidine decreased [Ca++]i and muscle force in parallel, whereas verapamil decreased [Ca++]i more strongly than muscle force. Clonidine (100 microM) inhibited the transient increase in [Ca++]i induced by CCh but not by caffeine (20 mM) in Ca++-free solution. Clonidine did not change the cAMP content in the presence of either 72.7 mM KCl or CCh. These results indicate that clonidine relaxes CCh-stimulated BTSM through the inhibition of CCh-induced increases in Ca++-influx, Ca++-release and intracellular signal transduction probably via imidazoline I1 receptors.

[Anaphylactic shock by ophthalmic wash solution containing chlorhexidine].

A 58 year-old male patient developed anaphylactic shock, possibly due to the use of chlorhexidine as ophthalmic wash solution. He was successfully resuscitated without any sequelae. The patient had increased levels of both histamine and tryptase. The skin test for allergy resulted in strong positive to chlorhexidine. There have been many reports regarding severe adverse reactions associated with use of chlorhexidine. It is necessary to pay attention to anaphylactic shock due to chlorhexidine.

The influence of volatile anesthetics on portwine stain.

To examine the relationship between volatile inhalation anesthetics and the fading of portwine stain (PWS), the study retrospectively investigated the incidence of termination of pulsed dye laser treatment vis-à-vis PWS fade during use of a general inhalation anesthetic in 107 infants and children with facial PWS. The fading of PWS is disadvantageous to the pulsed dye laser treatment. All patients received pulsed dye laser treatment under general anesthesia using one of four kinds of volatile inhalation anesthetics (halothane, enflurane, isoflurane, and sevoflurane). Two hours before the induction of anesthesia, all patients were rectally irrigated. Treatment was discontinued in none of the 44 patients in the halothane group or the 7 patients in the enflurane group; in one of the 29 in the isoflurane group; and in 10 of the 27 patients in the sevoflurane group. The sevoflurane group showed a significantly (P < 0.01) higher incidence of PWS fading. Thus, when using a volatile inhalation anesthetic in combination with pulsed dye laser treatment for PWS, caution should be exercised if sevoflurane is selected, as PWS fading is harmful to the pulsed dye laser.