Isoguvacine, isonipecotic acid, muscimol and N-methyl isoguvacine on the GABA receptor in rat sympathetic ganglia.

The GABA-mimetic activities of 4 analogues muscimol, isonipecotic acid, isoguvacine and N-methyl isoguvacine have been examined at the GABA receptor in the rat isolated superior cervical ganglion. The depolarizing action of all 4 analogues could be selectively antagonized by bicuculline methochloride and isopropyl bicyclophosphate. Muscimol was the only analogue more potent than GABA (molar potency ratio = 5.08 +/- 0.707). The potency of isoguvacine was 0.23 +/- 0.026 and isonipecotic acid 0.011 +/- 0.0028. N-methyl isoguvacine was less than 0.001 GABA.     

The action of conformationally restricted analogues of GABA onLimulus andHelix central neurones

Summary Intracellular recordings have been made from GABA sensitive neurones in the central nervous system ofLimulus andHelix. The following conformationally restricted analogues of GABA all possessed GABA-like activity onLimulus neurones andHelix excitatory GABA receptors: muscimol, thiomuscimol, THIP, isoguvacine and piperidine-4-carboxylic acid. It is suggested that GABA interacts with these receptors in a partially extended and almost planar conformation.     

Action of glutamic acid and of some glutamate analogues on the molluscan central neurones

Summary The effects of L-glutamic acid and of some glutamate analogues have been studied on the central nervous system of the snailHeobania vermiculata, using conventional electrophysiological techniques. The glutamate H-response had the mean equilibrium value of-(57±4) mV and was associated with a Cl^– conductance change. The D-response to glutamate application showed an involvement of sodium ions. Aspartate was agonist of glutamate action and displayed similar equilibrium value of the H-response, whereas quisqualate H-response was non-invertible.The authors wish to express their thanks to Prof. A. Curatolo for his continual interest and advice, and to Dr J. Stinnakre for the criticism of the text.     

A convulsant, 3-mercaptopropionic acid,decreases the level of GABA and GAD in rat pancreatic islets and brain

To investigate the properties of the gamma-aminobutyric acid (GABA) synthesizing enzyme, glutamate decarboxylase (GAD), in the brain and the pancreatic islets of the rat, GABA concentration in the brain and the pancreatic islets was measured after intraperitoneal administration of 3-mercaptopropionic acid (3-MP) at 25 mg/kg. 60 min after the administration of 3-MP, GABA concentration in the hypothalamus, the superior colliculus and the hippocampus of the brain decreased by 20–30% and in the pancreatic islets by 35%. The concentration in the pancreatic acini did not change. Western blotting showed that GAD activity in the pancreatic islets decreased after administration of 3-MP compared to the control. The activity of GAD in the pancreatic islets as well as brain can be modified by a convulsant, in this case 3-MP. These results suggest the properties of GAD may be similar in the pancreatic islets and brain.     

Transmural pressure as a determinant of basic intrinsic heart rate

Summary It was observed the heart rate was minimum at zero transmural pressure. The mean heart rate at zero transmural pressure was 23±5/min. This mean heart rate increased from 23±5/min to a peak value of 40±6/min (74% acceleratin) when the transmural pressure was raised from 0 to +4 mm Hg and to a similar peak value of 36±8/min (56% acceleration) when the transmural pressure was lowered from 0 to –4 mm Hg. The peak values attained at ±4 mm Hg were higly significant (p<0.001). It is concluded that the heart rate at zero transmural pressure represents the basic intrinsic pacemaker frequency independent of neural, humoral, thermal and haemodynamically induced mechanical influences.     

Reduction of bovine pulmonary hypertension by normoxia,verapamil and hexaprenaline

Summary In calves with hypoxic pulmonary hypertension, resistance was reduced by 40±3% with normoxia, 19±4% with verapamil, and 60±1% with hexoprenaline. It is possible that the increased resistance during normoxia is due partly to vasoconstriction rather than solely to vascular thickening, and that the vasoconstriction is due to an abnormality in calcium metabolism by the hypertensive vasculature.Acknowledgments. We thank Rosann Glas, Mary Munroe, Diann Smith and Linda Panepinto for technical assistance. This work was supported by the Colorado Heart Association and by the National Institutes of Health Grants HL-05973 and HL-29237. Verapamil (Isoptin) was supplied by Knoll Pharmaceutical Co. Hexoprenaline (ST 1512 HCl, Ipradol) was supplied by Österreichische Stickstoffwerke Linz.     

The number of neurons,glial and perineurium cells in an insect ganglion

Zusammenfassung Im zweiten Abdominalganglion der Indischen Stabheuschrecke (Carausius morosus Br.) wurden 600 ± 50 Neurone, 1000 ± 100 Gliazellen und 500 ± 30 Perineuriumzellen, zusammen etwa 2100 Zellen gezählt. Die Volumina des Ganglions, des Neuropilems und des Zellcortex betrugen etwa: 140 · 10^–4 mm³, 75 · 10^–4 mm³ und 65 · 10^–4 mm³.     

Effects of blood sampling,anesthesia and surgery on plasma vasopressin concentration in rats

The influence of blood sampling, anesthesia and surgery on plasma vasopressin concentration was assessed in rats. Mean plasma concentration in conscious, chronically catheterized rats was 1.4±0.1 pg/ml (n=6). This value remained constant over repeated plasma samplings in the same animals. On the other hand, decapitation increased the plasma vasopressin concentration to 6.0±2.4 (in pg/ml) (n=6), inactin anesthesia to 2.9±0.6 (n=6), anesthesia and femoral cannulation to 13.3±5.8 (n=6) and surgery for renal micropuncture to 81.3±35.0 (n=6). It is concluded that the level of circulating plasma vasopressin is highly dependent on the sampling technique and is closely related to the extent of surgery.     

The pharmacokinetics of VIP in dog and pig

Summary Pure porcine VIP was infused systemically in 4 conscious dogs and systemically and intraportally in 6 anesthetized pigs. At 2.3 pmoles·kg^–1·min^–1 the MCR was 10.7±1.0 ml·kg^–1·min^–1 in dog and 7.6±1.5 (systemic) and 16.5±2.0 (portal) in pig. The t 1/2's were 1.0±0.12 and 1.0±0.05 respectively. These values agree with those observed in man. This very high single pass tissue clearance does not suggest a hormonal role for VIP.     

GABA stimulates the rabbit corneal endothelial fluid pump.

GABA and its analogues were shown to activate the rabbit corneal endothelial fluid pump in the micromolar range. The stimulation was abolished by bicuculline and chlorpromazine, but not by picrotoxin.     

Asscorbic acid portects against male infertility in a teleost fish

An animal unable to synthesize ascorbic acid uniquely minicks human and non-human primates. Therefore, in this study we used the rainbow trout, a teleost fish, as the model animal to study the importance of dietary ascorbic acid on the fertilizing ability of sperm. A high concentration of ascorbic acid in semen plays a key role in maintaining the genetic integrity of sperm cells, by preventing oxidative damage to sperm DNA. This study will show that the concentration of asorbic acid in seminal plasma refelcts the dietary fed either an ascorbate-free diet (from 4.74±0.9 to 0.16±0.08 g ml^–1) or an ascorbate-rich diet (from 37.9±4.7 to 17.7± 3.2 g ml^–1) during the sperimnation season. The relationship between ascrobate status and fertility was studied in six groups of fish fed graded levels of ascorbic acid, which sperimated over a 150-day-period. Sperm from individual males was used to fertilize several batches of eggs. When the seminal plasma ascorbate concentration decreased to 7.3 g ml^–1 a significant decrease of fertilization rate and the hatching rate of embryos resulted. This is the first evidence that dietary ascorbate level directly affected sperm quality and influenced male fertility in a scruvy-prone vertebrate.     

Stability of ceftizoxime in aqueous solution

Summary Ceftizoxime, when assayed using the disc agar-diffusion method withBacillus subtilis ATCC 6633 as the indicator microorganism in buffered (pH 6) medium, retains its potency in aqueous solution at 4°C for about 3 months. Even after 4 months, some residual activity can be found. However, when this solution was assayed on the non-buffered medium, a seemingly quicker but misleading loss of potency was observed. This resulted from inadequate sensitivity of the assay. The sensitivity of the ceftizoxime assay was increased by lowering the incubation temperature from 37°C to 30°C. The pH- and incubation temperature-dependent potency must be kept in mind when assaying ceftizoxime.     

Oxidative modifications of low-density lipoproteins (LDL) by the human endothelial cell line EA.hy 926

Modifications of LDL by the EA.hy 926 cell line were compared to those generated by human umbilical vein endothelial cells (HUVEC). Thiobarbituric acid reactive substances (TBARS) index values (TBARS sample/TBARS cell-free control ratio) were 2.64±0.18 (m±SE, n=11) and 3.12±0.24 (n=11), for HUVEC and EA.hy 926, respectively. The percentage of the most electronegative modified LDL fraction (fraction C), assessed by using an ion-exchange chromatographic method based on fast protein liquid chromatography (FPLC), represented 14±3% (n=34) and 22±13% (n=10) of total modified LDL in HUVEC and EA.hy 926, respectively. LDL modified by both cell lines showed increased agarose electrophoretic mobility and apo B100 fragmentation on SDS-PAGE. None of the results were significantly different between the two cell lines. Superoxide anion production was 0.12±0.04 (n=11) and 0.07±0.01 nmol/min/mg cell protein (n=11) in HUVEC and EA.hy 926, respectively. Cell-specific effects on LDL were abrogated in cysteine-free medium. Moreover, cell-modified LDL were similarly degraded by J774 macrophage-like cells. We conclude that EA.hy 926 cells are a good model for investigating endothelial cell-induced modifications of LDL. Advantages include ready availability and less individual variability than with HUVEC.     

A simple,sensitive, non-stimulated photon counting system for detection of superoxide anion in whole blood

A simple, sensitive, non-stimulated assay was developed to measure the superoxide anion concentration in whole blood, using an ultra-sensitive chemiluminescence (CL) analyzer, and lucignin amplification. The assay system can be performed without leukocyte isolation or stimulant administration. The blood CL levels of healthy males (362.8±337.7 counts/10 sec) were not different from those of females (335±308.7 counts/10 sec) (p=0.64), whereas the CL levels in whole blood in patients with acute pancreatitis (2522±2014 counts/10 sec) were significantly higher than those of healthy controls (p<0.001). This assay system may be valuable in the future for quantitative measurement of reactive oxygen species in various disorders.     

Fatty acid modulation of antiestrogen action and antiestrogen-binding protein in cultured lymphoid cells

Summary Nonsteroidal antiestrogens reversibly and specifically inhibited the proliferation of two estrogen receptornegative lymphoid cell lines (EL4 and Raji) in a dose-dependent manner. [³H]Thymidine incorporation of concanavalin A-stimulated primary splenocytes was also inhibited by 10^–6 M clomiphene (1-[4-(2-diethylaminoethoxy)phenyl]-1,2-diphenyl-2-chloroethylene). The antiproliferative effect could be prevented by the simultaneous presence in the growth medium of 10^–5 M linoleic acid or 10^–5 M arachidonic acid but not by 10^–6 M estradiol. Both lymphoid cell lines had high affinity antiestrogen-binding sites whose affinity could be altered by conditions of growth. Growth of EL4 cells in RPMI 1640 medium supplemented with charcoal-pretreated 5% fetal calf serum (charcoal-stripped medium) resulted in significantly higher affinity (K_d 0.54 nM±0.11 nM; n=6) than growth in medium supplemented with untreated serum (complete medium) (K_d=1.68 nM±0.48 nM; n=6) (p<0.001). This change in affinity was partly due to removal of fatty acids from the growth medium by charcoal pretreatment, since addition of 10^–5 M linoleic acid or 10^–5 M gamma-linolenic to charcoal-stripped medium decreased the affinity of the antiestrogen-binding protein. In contrast, growth in 10^–5 M stearic acid or 10^–5 M oleic acid did not significantly alter the affinity of the antiestrogen-binding protein, whereas 10^–5 M palmitic acid significantly increased its affinity. The same fatty acids were also tested for their intrinsic effects on EL4 cell proliferation. Oleic, linoleic and gamma-linolenic acids were growth stimulatory while stearic and palmitic acids were not. Thus linoleic and gamma-linolenic acids whose presence in the growth medium was associated with decreased affinity of [³H]tamoxifen (1-[4-(2-dimethylaminoethoxy)phenyl]-1,2-diphenylbut-1(Z)-ene) binding to the intracellular antiestrogen-binding protein were also growth stimulatory. Unsaturated fatty acids have previously been shown to inhibit binding of [³H]tamoxifen to the antiestrogen-binding protein in a cell-free system. The present observations demonstrate that unsaturated fatty acids also modify the affinity of the antiestrogen-binding protein in intact cells.     

Lipopeptaibols, a novel family of membrane active, antimicrobial peptides

Lipopeptaibols are members of a novel group of naturally occurring, short peptides with antimicrobial activity, characterized by a lipophilic acyl chain at the N-terminus, a high content of the turn/helix forming α-aminoisobutyric acid and a 1,2-amino alcohol at the C-terminus. The amino acid sequences range from 6 to 10 residues and the fatty acyl moieties from 8 to 15 carbon atoms. The peptide portion of lipopeptaibols can be shorter than those of the nonlipidated peptaibols that range from 10 to 19 amino acid residues. The longest peptides fold into a mixed 3₁₀/α helix, whereas the shortest peptides tend to adopt a β-turn/sheet structure. Using solution methodologies, a series of analogues of trichogin GA IV was synthesized which allowed determination of the minimal lipid chain and peptide main-chain lengths for the onset of membrane activity and exploitation of a number of spectroscopic techniques aimed at determining its preferred conformation under a variety of conditions and investigating in detail its mode of interaction with, and its effect on, the phospholipid membranes. Received 26 January 2001; received after revision 7 March 2001; accepted 15 March 2001     

GABA stimulates the rabbit corneal endothelial fluid pump

Summary GABA and its analogues were shown to activate the rabbit corneal endothelial fluid pump in the micromolar range. The stimulation was abolished by bicuculline and chlorpromazine, but not by picrotoxin.This work was supported by PHS-NIH Grant No. 1R01 EY 00965, to S.D.     

α1-Adrenergic stimulation of ketogenesis and fatty acid oxidation is associated with inhibition of lipogenesis in rat hepatocytes

Summary The effect of norepinephrine on fatty acid synthesis (³H₂O incorporation into fatty acids), on fatty acid oxidation to CO₂ and on ketogenesis was studied in isolated hepatocytes of fed rats. After incubation with norepinephrine (50 M), lipogenesis was lower (5.7±1.1 nmoles³H₂O incorporated into fatty acids/mg dry weight/30 min) than in controls (7.5±1.7; n=6, p<0.02). In contrast, (1-¹⁴C) palmitate conversion into total ketone bodies was increased to 10.9±1.8 nmoles/mg/30 min with norepinephrine, vs 8.5±1.6 in controls (p<0.05), and more (1-¹⁴C) palmitate was converted to¹⁴CO₂ with norepinephrine than in controls (1.48±0.10 nmoles/mg/30 min vs 1.06±0.11, p<0.05). The inhibitory effect of norepinephrine on lipogenesis was abolished by addition of the ₁-receptor blocker prazosin, but not by ₂ or -blockers. The results demonstrate that the ketogenic effect of norepinephrine is coupled with an inhibitory effect on lipogenesis which may be explained by diminished activity of acetyl-CoA carboxylase, diminished formation of malonyl-CoA and decreased activity of carnitine palmitoyl transferase I.     

The conduction velocity,number and diameter of unmyelinated fibers in Remak's nerve

Summary Remak's nerve in the chicken was examined ultrastructurally and electrophysiologically to determine the characteristics of fibers in the nerve trunk. The ratio of unmyelinated fibers to myelinated ones was 1111. The mean number of unmyelinated fibers was 3555±232 (SEM, n=5) and they had a mean diameter of 0.502±0.034 (SEM) m. The compound action potential consisted almost entirely of a large diphasic waveform which had a mean peak conduction velocity of 0.62±0.031 (SEM, n=5) m·s^–1 at 37°C.     

Totally synthetic (±)-13-Alkyl-3-hydroxy and methoxy-gona-1,3,5(10)-trien-17-ones and related compounds

Zusammenfassung Von mehreren (±) 3-Oxy- und (±) 3-Methoxy-13-alkylgona-1,3,5(10)trien-17-onen und verwandten Verbindungen, einschliesslich von Vertretern der (±) 13-Alkylgon-4-en-3-on-Reihe, werden Totalsynthese und biologische Wirksamkeit beschrieben.