Acute response of the arterial wall to pulsed laser irradiation

This study was designed to examine the acute response of normal arterial wall to pulsed laser irradiation. Irradiation with an Excimer or a Holmium YAG laser was performed in 15 normal iliac sites of 8 male New Zealand white rabbits. The excimer laser was operated at 308 nm, 25 Hz, 50 mj/mm²/pulse, and 135 nsec/pulse and the Ho:YAG laser was operated at 2.1 μm, 3.5 Hz, 400 mj/ pulse, 250 μsec/pulse. The excimer and Ho:YAG laser were coupled into a multifiber wire-guided catheter of 1.4 and 1.5 mm diameter, respectively. The mean luminal diameter increased similarly from 2.01 ± 0.29 to 2.46 ± 0.27 mm (P < 0.0005) and from 2.09 ± 0.53 to 2.45 ± 0.30 mm (P < 0.005) after excimer and Ho:YAG laser irradiation, respectively. Perforation occurred in 3 of 15 Ho:YAG irradiated sites and 0 of 15 excimer laser irradiated sites. The sites irradiated with excimer or Ho:YAG laser had similar histologic features, consisting of shedding of the endothelium, disorganization of internal elastic lamina, localized necrosis of vascular smooth muscle cells, and fissures in the medial layer. However, the sites irradiated with excimer laser had lower grading scores than those irradiated with the Ho:YAG laser (P<0.05). Irradiation with excimer or Ho:YAG laser of normal arteries results in: (1) vasodilation of the irradiated artery; (2) localized mechanical vascular injury, and (3) Ho:YAG laser induces more severe damage to the arterial wall than excimer. © 1993 Wiley-Liss, Inc.     

Selective Effect of Adjuvant Arthritis on the Disposition of Propranolol Enantiomers in Rats Detected Using a Stereospecific HPLC Assay

Nonstereospecific studies have indicated that the pharmacokinetics of propranolol (PR) are altered in inflammatory conditions such as arthritis. However, as the kinetics and dynamics of PR are stereoselective, we examined the effect of adjuvant arthritis (AA) on the disposition of the individual enantiomers. A novel normal-phase stereospecific HPLC assay for PR was developed involving chiral derivatization with S-(naphthyl)ethyl isocyanate and fluorescence detection. Oral and iv doses of racemic PR were administered to control and AA rats (n = 6). AA had no significant effect on either clearance or S:R ratio after iv doses. On the other hand, after oral doses, clearance was significantly decreased in AA. Although significant for both enantiomers, this effect was more pronounced on the less active R-enantiomer. The AUC R:S ratio was, therefore, significantly altered (AA, 14 ± 3.0; control, 4.3 ± 1.2). Increased total (S + R) plasma concentrations of PR in AA, possibly due to a reduced intrinsic clearance, therefore, reflect mainly increased concentrations of the less active R-enantiomer.     

Analysis of the Inhibitory Effect of Oestradiol on Functional GABA/5-HT Relationship in the Rat Suprachiasmatic Area

The purpose of this study was to test the capacity of oestradiol to modulate the stimulating effect of a-aminobutyric acid (GABA) on serotonin (5-HT) metabolism, previously described in the Suprachiasmatic area of the male rat. After an in vivo stimulation of GABA transmission by systemic administration of a GABA-transaminase inhibitor (amino-oxyacetic acid) or a GABA_B agonist (RS-baclofen), the 5-HT metabolism was studied in the Suprachiasmatic area of ovariectomized, and ovariectomized oestradiol-treated rats. Amino-oxyacetic acid or RS-baclofen treatment increased the endogenous content of 5-HT in the Suprachiasmatic area of males and ovariectomized rats. These two treatments were without effect in ovariectomized oestradiol-treated rats. GABA transmission stimulation induced by amino-oxyacetic acid treatment failed to affect the release and synthesis of 5-HT in ovariectomized oestradiol-treated rats while it increased these two parameters of 5-HT metabolism in the Suprachiasmatic area of male and ovariectomized rats. To investigate the main target of oestradiol effect, comparative studies of the serotoninergic and GABAergic metabolism in the Suprachiasmatic area were performed in the three experimental groups. Under our experimental conditions the endogenous 5-HT metabolism was similar between ovariectomized and ovariectomized oestradiol-treated rats. Nevertheless, 5-HT metabolism was higher in the two female groups than in the male group. Neither GABA metabolism nor GABAergic response to GABA-related drug treatment differed between ovariectomized, and ovariectomized oestradiol-treated rats. However, the turnover of GABA was higher when compared to the two female groups. It is concluded that the lack of 5-HT responsiveness to GABA transmission stimulation in ovariectomized oestradiol-treated rats was not related to an effect of oestradiol on 5-HT metabolism or to an effect of the steroid on GABA turnover. Furthermore, our results suggest a sex difference in the activity of serotoninergic and GABAergic systems in the Suprachiasmatic area.     

Patterns of C-heterochromatin and telomeric DNA in two representative groups of small apes,the genera <Emphasis Type="Italic">Hylobates</Emphasis> and <Emphasis Type="Italic">Symphalangus</Emphasis>

The course of chromosome evolution in small apes is still not clear, though painting analyses have opened the way for elucidating the puzzle. Even the C-banding pattern of the lar-group of gibbons (the genus Hylobates) is not clarified yet, although our previous studies suggested that lar-group gibbons have a unique C-banding pattern. We therefore made observations to establish C-banded karyotypes of the agile gibbons included in the lar-group. The data were compared with those of siamangs (the genus Symphalangus), which carry distinctive C-bands, to determine the chromosomal patterns in each group. C-banded chromosomes of agile gibbons showed several terminal, interstitial and paracentric bands, whose patterns are specific for each chromosome, whereas the C-bands of siamangs were located only at the terminal and centromeric regions in most chromosomes. Moreover, the C-bands of agile gibbons and siamangs were shown to be G+C-rich and A+T-rich DNA, respectively, by DAPI/C-band sequential staining. Additionally, PRINS labelling with a telomere primer revealed that agile gibbons have telomeric DNA only at chromosome ends where there is no C-band (non-telomeric heterochromatin), whereas the telomeric DNA of siamangs is located in the terminal C-banded regions (telomeric heterochromatin). Although the evolutionary mechanisms in small apes are still unknown, C-banding patterns and distribution of telomeric DNA sequences should provide valuable data to deduce the evolutionary pathways of small apes.     

Killer DNA plasmids of the yeast Kluyveromyces lactis

Summary We have studied the structure of the two linear DNA plasmids, kl and k2, present in killer strains of Kluyveromyces lactis. Two killer strains of different origins, CBS 2359 and IFO 1267 were examined. For both strains, identical restriction maps of kl and k2 DNA were obtained. Several restriction sites previously reported for the kl DNA of the strain IFO 1267 have been confirmed. The molecular weights of these double-stranded DNAs were 8.8 kilobase pairs for kl and 13.4 for k2, as determined by electrophoresis of restriction fragments. The plasmid DNA from a nonkiller mutant, NK2/1, was also examined. In this mutant, the kl DNA was replaced by a smaller DNA (5.9 kilobase pairs), the k2 DNA being normal. Restriction enzyme analysis showed that the new plasmid DNA was also linear. Hybridization experiments demonstrated that it was derived from the kl DNA by deletion of a 2.9 kilobase pair segment from the central part of the kl DNA. The deleted segment carries a gene involved in toxin production, but is not related to immunity since the mutant is resistant to killers. The plasmid DNA of K. lactis showed no detectable sequence homology with the double stranded RNA of the killer system of Saccharomyces cerevisiae. Neither was any homology found with nuclear and mitochondria) DNA.     

The radioimmunoassay of histamine release from circulating basophils in an in vitro study as support for the in vivo active systemic anaphylaxis test in the guinea pig

An in vitro anaphylaxis test is described which explores the ability of compound re-challenge to induce histamine release from polynuclear basophils in whole blood of guinea pigs that had previously been sensitised. This test is used in drug safety, in support of the in vivo active systemic anaphylaxis test, which is sometimes required by regulatory authorities, when the observed clinical signs are not conclusive. This sensitive and discriminative test is inexpensive and reduces animal utilisation.     

In vivo evidence for acetylcholine control of serotonin release in the cat caudate nucleus: influence of halothane anaesthesia

Using a push-pull cannula technique and an isotopic method for the estimation of [3H]serotonin continuously synthesized from [3H]tryptophan, the effects of acetylcholine were investigated on the in vivo release of [3H]serotonin in the cat basal ganglia and the dorsal raphe nucleus. The unilateral striatal application of acetylcholine (5 x 10(-5) M) reduced local release of [3H]serotonin. This effect was mimicked by nicotine (5 x 10(-5) M) and prevented by mecamylamine (10(-6) M. Oxotremorine (5 x 10(-5) M) had no effect on the local release of [3H]serotonin. All these treatments failed to modify [3H]serotonin release in the ipsilateral substantia nigra or in the dorsal raphe nucleus. The superfusion of serotonergic nerve terminals of the caudate nucleus with tetrodotoxin prevented the inhibitory acetylcholine-induced effect on serotonin release. Furthermore, bicuculline (5 x 10(-5) M) in the caudate nucleus blocked the effect of nicotine, while gamma-aminobutyric acid (10(-5) M) induced a decrease in local release of [3H]serotonin. These data strongly suggest that the inhibitory control exerted by acetylcholine on serotonergic transmission could involve gamma-aminobutyric acid interneurons. Acetylcholine-induced changes in [3H]serotonin release were only observed in non-anaesthetized "encéphale isolé" cats and not in halothane-anaesthetized animals. The possibility that such a regulation could be presynaptic (direct or through other neurotransmitters) or related to a change in the activity of the serotonergic raphe-striatal neuronal system is discussed.     

In vitro H-serotonin (5-HT) synthesis and release in BALBc and C57BL mice. II. Cell body areas

"In vitro" 3H-5-HT and 3H-5-HIAA, newly synthesized from 3H-TRP, are measured in the brainstem, the anterior raphe nuclei and the locus coeruleus of C57BL and BALBc mice. As found for the caudate nucleus and the hippocampus, higher synthesis and release are determined in C57BL than in BALBc, for the locus coeruleus and more globally, for the brainstem. But these differences disappear when the study is carried on the raphe dorsalis and the raphe centralis nuclei. Therefore the serotonergic activity could be independently regulated at the level of cell bodies and at those of terminals. However, the 5-HT metabolism of NRD of BALBc mice could be submitted to a specific autoinhibition which could explain a lower 5-HT synthesis and release at the corresponding terminal level, compared to C57BL.     

In vitro H-serotonin (5-HT) synthesis and release in BALBc and C57BL mice. I. Terminal areas

"In vitro," 3H-5-HT and 3H-5-HIAA newly synthesized from 3H-TRP are measured in the caudate nucleus and the hippocampus of C57BL and BALBc mice. Higher synthesis, utilization and release are to be found in C57BL than in BALBc strain. In the hippocampus of C57BL this higher synthesis is due both to higher tryptophan hydroxylase activity and to higher tryptophan uptake ability. But in the caudate nucleus the initial accumulation of tryptophan is similar in both strains. Finally the two forms of monoamine oxidase (A and B) show also similar activities in both strain. These data will be compared to those obtained at the nerve cell body level in the paper (II).