Novel tricyclic-alpha-alkyloxyphenylpropionic acids: dual PPARalpha/gamma agonists with hypolipidemic and antidiabetic activity.

Synthesis and structure-activity relationships of tricyclic alpha-ethoxy-phenylpropionic acid derivatives guided by in vitro PPARalpha and PPARgamma transactivation data and computer modeling led to the identification of the novel carbazole analogue, 3q, with dual PPARalpha (EC(50) = 0.36 microM) and PPARgamma (EC(50) = 0.17 microM) activity in vitro. Ten days treatment of db/db mice with 3q improved the insulin sensitivity, as measured by OGTT, better than that seen with both pioglitazone and rosiglitazone treatment, suggesting in vivo PPARgamma activity. Likewise, 3q lowered plasma triglycerides and cholesterol in high cholesterol fed rats after 4 days treatment, indicating in vivo PPARalpha activity. Investigations of the pharmacokinetics of selected compounds suggested that extended drug exposure improved the in vivo activity of in vitro active compounds.

The nitric oxide synthase inhibitor, L-NAME, block phencyclidine-induced disruption of prepulse inhibition in mice.

RATIONALE: Schizophrenia is a major public health problem that affects approximately 1% of the population worldwide. Schizophrenia-like syndromes can be induced in humans by phencyclidine (PCP), a drug with marked psychomimetic properties. Recent studies show that the behavioural and biochemical effects of PCP in rats are blocked by nitric oxide synthase (NOS) inhibitors, suggesting that NO plays an important role in the pharmacological effects of PCP. OBJECTIVE: The aim of this study was to investigate if PCP-induced disruption of prepulse inhibition of acoustic startle could be blocked by the NOS inhibitor, L-NAME, in mice. RESULTS: The present study shows that PCP readily disrupts prepulse inhibition in mice normally without affecting pulse-alone trials. Furthermore, L-NAME blocked the PCP-induced disruption of prepulse inhibition in a dose-related manner. CONCLUSIONS: The PCP-induced disruption of prepulse inhibition and the ability of L-NAME to block this effect in both rats and mice suggest that this is a general and not a species-specific effect. The results of the present study further suggest that PCP exerts at least some of its actions in the central nervous system by a NO-dependent mechanism.

Role of dopamine in prepulse inhibition of acoustic startle.

RATIONALE: Prepulse inhibition of acoustic startle is the reduction in startle response to an intense auditory stimulus when this stimulus is immediately preceded by a weaker prestimulus. Prepulse inhibition occurs normally in humans and experimental animals, but schizophrenic persons often exhibit a marked impairment in this measure. Previous studies have shown that dopamine (DA)-dependent neuronal mechanisms are involved in the modulation of prepulse inhibition. OBJECTIVE: Experiments were conducted in rats to elucidate further the involvement of DA-ergic mechanisms in prepulse inhibition. RESULTS: In line with previous studies, the indirect DA agonist, amphetamine, was shown to decrease prepulse inhibition. A close reverse relationship over time between DA overflow in the nucleus accumbens and prepulse inhibition was obtained using a technique allowing concomitant measurement of these parameters in awake, freely moving rats. This effect was more pronounced in amphetamine-treated rats compared to rats treated with equimolar doses of cocaine, which increased DA overflow without affecting prepulse inhibition. In other experiments, the combined treatment with subthreshold doses of the selective DA D1 agonist, SKF 38393, and the selective DA D2 agonist, quinpirole, was also shown to decrease prepulse inhibition. Finally, the selective DA D2 antagonist, raclopride, was shown to enhance prepulse inhibition. CONCLUSIONS: In line with previous studies, it is concluded that DA neurotransmission is involved in the modulation of prepulse inhibition and that the ventral part of the mesostriatal DA system may serve an important role in this modulation. Furthermore, the possibility is discussed that the discrepant results on prepulse inhibition obtained with amphetamine and cocaine may disclose functionally relevant differences in their mechanisms of action, and that the enhancement of prepulse inhibition induced by some antipsychotics in rats may reflect their propensity to induce adverse mental effects in humans.

Determination of estramustine phosphate and its metabolites estromustine, estramustine, estrone and estradiol in human plasma by liquid chromatography with fluorescence detection and gas chromatography with nitrogen-phosphorus and mass spectrometric detection.

Bioanalytical methods for the determination of estramustine phosphate by liquid chromatography and its four main metabolites estromustine, estramustine, estrone and estradiol by gas chromatography are described. For the estramustine phosphate assay the plasma was purified by protein precipitation followed by a C18 solid-phase extraction. For the metabolite assay the plasma samples were purified by a C18 solid-phase and liquid-liquid extraction procedure and derivatised by silanization. Thereafter, estramustine and estromustine were quantified by gas chromatography with nitrogen-phosphorus detection and estradiol and estrone were quantified by gas chromatography with selected ion monitoring. The methods were validated with respect to linearity, selectivity, precision, accuracy, limit of quantitation, limit of detection, recovery and stability. The limit of quantitation was 2.3 micromol/l for estramustine phosphate, 30 nmol/l for estromustine and estramustine, 12 nmol/l for estrone and 8 nmol/l for estradiol. The results showed good precision and accuracy for estramustine phosphate and the four metabolites. The intermediate precision was 6.2-13.5% (C.V.) and the accuracy was 91.8-103.9%.

How to obtain hemostasis after aortic surgery.

BACKGROUND: The establishment of hemostasis without the excessive transfusion of homologous blood and blood products is critical to successful aortic surgery. METHODS AND RESULTS: By using preoperative autologous blood donation and intraoperative blood conservation measures, 85% of patients can undergo aortic surgical procedures without homologous blood or product transfusions, and almost three-quarters of patients will still not have required homologous transfusions by the time of discharge. In contrast, three-quarters of those patients who cannot donate blood preoperatively will require homologous blood transfusions. CONCLUSIONS: The strategy described is safe: our overall survival rate for 204 patients has been 98%, with a 1% incidence of stroke.

Active suppression in orally tolerized rats coincides with in situ transforming growth factor-beta (TGF-beta) expression in the draining lymph nodes.

Adult rats were fed pellets containing ovalbumin (OvA) during 4 weeks, and were 2 weeks thereafter immunized subcutaneously with a mixture of OvA and human serum albumin (HSA) in Freund's complete adjuvant (day 0). As a result of the immunization, the draining lymph nodes of the nontolerized (control) rats were heavily enlarged from day 10 to day 18; however, this size increase was absent in the OvA-fed rats. This manifestation of active suppression in the tolerized rats was preceded by the appearance of scattered CD4+ TGF-beta-expressing T cells in the T cell area of their lymph nodes (days 5-8); correspondingly, the levels of TGF-beta mRNA in the nodes were elevated in the tolerant rats compared with the control rats. The anti-OvA antibody levels in sera from the rats revealed that there was an initial B cell priming in the OvA-fed group, with levels higher than in the control group during the first week. Thereafter, suppression governed the response, and from day 10 onwards the anti-OvA levels were considerably lower than in the controls. When other groups of animals were pretreated with neutralizing anti-TGF-beta antibodies 1 day before the immunization, the anti-OvA response of the OvA-fed rats was restored to the levels of the control group, demonstrating the importance of TGF-beta in the maintenance of suppression. In conclusion, we demonstrate that TGF-beta-producing cells appear in the draining lymph nodes shortly after immunization in rats made orally tolerant using a relatively high-dose feeding regime; these cells are probably responsible for the down-regulation of the immune response observed in the OvA-fed rats.

Involvement of the medial geniculate body in prepulse inhibition of acoustic startle.

Prepulse inhibition of acoustic startle is the normal reduction in startle response to an intense auditory stimulus when this stimulus is immediately preceded by a weaker prestimulus. Previous studies have shown that several neuroanatomical structures and pathways in the brain are involved in the modulation of prepulse inhibition. In the present study, the functional importance of the medial geniculate body (MG) in the modulation of prepulse inhibition was investigated. To this end, in vivo brain microdialysis probes were used to infuse drugs locally into the MG of awake, freely moving rats simultaneously with startle response and prepulse inhibition measurements in the same animals. Intrageniculate infusion of the sodium channel blocker, tetrodotoxin, significantly reduced prepulse inhibition without affecting baseline startle amplitude. A similar effect was obtained after intrageniculate infusion of the GABA(B) receptor agonist, baclofen. In addition, intrageniculate infusion of muscimol, an agonist at the GABA(A) receptor complex, reduced prepulse inhibition, although this effect was obtained at a higher concentration of the drug compared to that of baclofen. These studies suggest that the MG is involved in the modulation of prepulse inhibition and that auditory signals relayed via the MG may be subjected to inhibitory control at this level, involving GABA neurotransmission.

Crystallization and preliminary X-ray analysis of birch-pollen allergen Bet v 1 in complex with a murine monoclonal IgG Fab' fragment.

The human type I allergic response is characterized by the presence of allergen-specific serum immunoglobulin E (IgE). Allergen-mediated cross-linking of receptor-bound IgE on the surface of mast cells and circulating basophils triggers the release of mediators, resulting in the development of the clinical symptoms of allergy. In order to study the structural basis of allergen-antibody interaction, a complex between the major birch-pollen allergen Bet v 1 and a Fab' fragment isolated from the murine monoclonal Bet v 1 antibody BV16 has been crystallized. Complex crystals belong to space group P1, with unit-cell parameters a = 91.65, b = 99.14, c = 108.90 A, alpha = 105.7, beta = 98.32, gamma = 97.62 degrees, and diffract to 2.9 A resolution when analyzed at 100 K using synchrotron-generated X--rays.

Repeated administration of amphetamine induces sensitisation to its disruptive effect on prepulse inhibition in the rat.

Male Sprague-Dawley rats were repeatedly treated with amphetamine (AMP, 1 mg/kg, SC) at 3- day intervals for 15 days and tested for prepulse inhibition of acoustic startle after each treatment. This treatment regimen induced sensitisation in the animals as evidenced by a progressive increase in the disruptive effect of AMP on prepulse inhibition. Persistent changes in brain function was indicated, since an increase in disruptive effect was observed in sensitised animals also after a 22-day-long drug- and test-free period. The development of sensitisation was blocked by pretreatment with haloperidol (HPD, 0.1 mg/kg, SC), which suggests that sensitisation to the disruptive effect of AMP was dependent on dopamine (DA) D2 receptor activation. Furthermore, the development of sensitisation was blocked by adrenalectomy, which suggests that sensitisation was dependent also on circulating adrenal hormones. Increased DA-ergic activity has been implicated in the pathophysiology of schizophrenia and AMP-induced sensitisation to the neuronal functions that modulate prepulse inhibition may be an experimental model to investigate this hypothesis.

Effect of transurethral microwave thermotherapy. An evaluation with MR imaging.

PURPOSE: To detect morphological changes in the prostate, as depicted with MR imaging, in order to clarify the effects of transurethral microwave thermotherapy (TUMT). MATERIAL AND METHODS: Twenty patients with prostatism and a prostatic volume of 30-71 cm3 underwent MR examination before, the day after, and 6 months after treatment. TUMT was carried out with a Prostatron. A method to detect oedematous changes on heavily T2-weighted MR images was developed and used as an indicator of morphological changes. RESULTS: The study showed some correlation (r=0.59) between the energy given at TUMT and an increased T2 signal. All patients with increased T2 signal except one were found among those who received the highest amount of energy to the prostate. Of 8 patients, 6 showed a symptomatic response to the treatment and 2 did not. There was a weak statistical correlation (r=0.41) between treatment response and increased T2 signal. CONCLUSION: The study does not support the view that TUMT leads to significant necrosis in the prostate with loss of tissue and retraction. We theorize that the response to TUMT may be caused by a denervation of the prostate.

Prepulse inhibition of acoustic startle, a measure of sensorimotor gating: effects of antipsychotics and other agents in rats.

Schizophrenic patients are deficient in various neurologic measures reflecting information processing. One such measure in prepulse inhibition (PPI) of acoustic startle, in which schizophrenics display less inhibition than normal subjects. PPI is also diminished in rats treated with psychotomimetic drugs such as amphetamine and phencyclidine. PPI has been suggested as a model relevant for studying the pathophysiology of schizophrenia. We studied the effect of a variety of antipsychotics and putative antipsychotics and some key reference compounds on the acoustic startle response (ASR) and PPI. Some, but not all, antipsychotics tested (mainly selective dopamine D2 antagonists) enhanced PPI. Remoxipride and clozapine, both of which are antipsychotics, and the very potent and highly selective D2 antagonist, NCQ-298, did not. It is concluded that enhanced PPI in otherwise untreated rats does not reflect antipsychotic efficacy. We further noted that the effect on PPI was independent of the effect on ASR.

Changes in the acoustic startle response and prepulse inhibition of acoustic startle in rats after local injection of pertussis toxin into the ventral tegmental area.

The effect of local injection of pertussis toxin (PTX) into the ventral tegmental area (VTA) on acoustic startle in rats was investigated. The PTX treatment caused only minor effects of its own on the acoustic startle response (ASR) or prepulse inhibition (PPI) of acoustic startle. However, systemic treatment with the indirect DA receptor agonist, amphetamine (2 mg/kg, SC) caused a significant increase in ASR magnitude and a significant disruption of PPI in PTX-treated rats while no such effects were observed in sham-treated rats. Treatment with the direct DA receptor agonist, apomorphine (2 mg/kg, SC), caused a significant disruption of PPI, an effect that was observed in both PTX- and sham-treated rats. Treatment with the 5-HT1A receptor agonist, 8-OH-DPAT (0.5 mg/kg, SC), did not affect PPI in either group but caused a marked increase in ASR magnitude in sham-treated rats. Interestingly, this effect was blocked in PTX-treated rats. The present results suggest that local injection of PTX into the VTA causes an increased sensitivity to the behavioural effects of psychostimulants on acoustic startle and may also suggest that intact midbrain 5-HT1A receptors are essential for the effect of 5-HT1A agonists on acoustic startle.

The atypical antipsychotic, remoxipride, blocks phencyclidine-induced disruption of prepulse inhibition in the rat.

The effect of various typical (haloperidol) and atypical (clozapine, raclopride, remoxipride) antipsychotics on phencyclidine (PCP)-induced disruption of sensorimotor gating was tested in rats using an acoustic startle paradigm. Clozapine (4-40 mumol/kg), haloperidol (1-5 mumol/kg) and raclopride (1-12 mumol/kg) failed to reverse PCP-induced disruption of prepulse inhibition (PPI) of the acoustic startle response. In contrast, remoxipride (12-60 mumol/kg) caused a dose-dependent block of this effect. PCP-induced disruption of PPI is a widely accepted animal model of a corresponding behavioural deficit observed in schizophrenia although little evidence has been presented that it is in fact sensitive to antipsychotic agents. The present results indicate that remoxipride behaves in a unique way in this model compared to clozapine, haloperidol and raclopride.

Stability of vitamins in soybean oil fat emulsion under conditions simulating intravenous feeding of neonates and children.

The stability of the fat-soluble vitamins and thiamine, riboflavin, pyridoxine, ascorbate, and folic acid when administered as part of Intralipid 10% was evaluated. Intravenous pediatric vitamin formulations (Vitalipid N Infant and Soluvit Infant) containing the daily dose for term neonates were diluted with Intralipid to final volumes of 12.5 mL or 260 mL. The volumes were then delivered for 24 hours via dihexylethyl-phthalate-free in vitro infusion systems that simulated clinical conditions. The first vitamin-Intralipid admixture (12.5-mL volumes) was pumped (by syringe) during 24 hours through neonatal infusion sets placed in an incubator (37 +/- 1 degree C, bilirubin light). Aliquots were collected at the beginning of the experiment and after 24 hours of infusion. The second admixture (260-mL volumes) was pumped (by a peristaltic pump) through infusion sets during 24 hours and was exposed to periodic indirect sunlight and to continuous fluorescent light to stimulate the feeding of a child. Aliquots were collected at 0, 8, and 24 hours during administration of the admixtures. The vitamin concentrations were measured by high-performance liquid chromatography. The effect of the vitamins on the stability of the emulsion was also assessed by visual inspection, lipid globule size distribution, and pH change. Losses of vitamins vary from different experimental conditions and were for the fat-soluble vitamins essentially nil, except for phylloquinone, which was 5% to 17% below the initial level. Thiamine, pyridoxine, and folic acid were stable. Losses of riboflavin and ascorbate amounted to 10% to 20% and 9% to 52%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of ageing on extracellular ascorbate concentration in rat brain.

In vivo voltammetry at electrochemically pretreated carbon fibre electrodes was used to investigate the effect of ageing on extracellular ascorbate (AA) concentration in the rat brain. Recordings from the nucleus accumbens in 3-, 6- and 18-month-old Sprague-Dawley rats revealed an age-related decrease in basal extracellular AA concentration. The mean AA current measured in 18-month-old rats was less than 10% of the current measured in 3-month-old rats. Systemic administration of ethanol (1.0 g/kg, i.p.) caused an increase in the AA signal measured in this area in all 3 age groups tested. However, the effect on AA was significantly less pronounced in 18-month-old rats. Further analysis of the AA signal revealed a gradual increase in AA release during terminal anoxia. Also in this case the effect on AA was significantly less pronounced in 18-month-old rats. This difference was also observed in the caudate putamen, another dopamine (DA) rich area in the brain. No significant difference in AA release was observed in the frontal cortex where the DA concentration is low. The increase in AA was followed by a pronounced increase in extracellular DA in the nucleus accumbens and caudate putamen. This release of DA was accompanied by a prompt reversal of the AA signal possibly explained by a DA-dependent autoxidation of AA. These results suggest a role for brain AA in the process of ageing.

Evidence for a dissociation between cardiovascular and behavioral reactivity in the spontaneously hypertensive rat.

Spontaneously hypertensive rats of the Okamoto strain (SHR) were compared with normotensive rats of the Wistar-Kyoto strain (WKY) on the acoustic startle response in rats prepared for simultaneous blood pressure recordings. Blood pressure was continuously recorded by means of an indwelling cannula in the caudal tail artery. The presentation of the startle stimulus caused a blood pressure response in both strains consisting of an initial increase in blood pressure followed by a decrease and after that a longer lasting, but less pronounced second increase in pressure. The startle-elicited increase in blood pressure was significantly elevated in SHRs and at the same time the acoustic startle response was depressed as compared to WKY rats. These data indicate a dissociation between cardiovascular and behavioral reactivity in the SHR.

Behavioral reactivity in spontaneously hypertensive rats.

Spontaneously hypertensive rats of the Okamoto strain (SHR) were compared with inbred normotensive rats of the Wistar-Kyoto strain (WKY) and with normally bred Wistar rats (NT) in tests on the audiogenic immobility reaction (freezing), open-field behavior in a dark and an enlightened arena respectively, auditory startle response and male sexual behavior. Compared to the WKYs the SHRs showed increased locomotion and rearing in the open-field situations, reduced startle response and shortened immobility reaction. The SHRs differed in the same way from the NT rats with the exception for motor activity in the dark arena, where no differences were observed. The WKY rats showed less motor activity than the NT animals. Both SH and WKY rats showed shorter latency time for ejaculation than the NT rats. The characteristics of the behavior patterns displayed by the SH rats were interpreted as indicating a reduced propensity for fear reactions in this strain of rats compared to the WKY and NT strains used in the present study.

Influence of dietary partially hydrogenated vegetable and marine oils on membrane composition and function of liver microsomes and platelets in the rat.

The aim of the present study was to investigate the influence of partially hydrogenated vegetable and marine oils on membrane composition and function of liver microsomes and platelets with particular reference to the metabolism of linoleic acid and the production of arachidonic acid metabolites. Four groups of male weanling rats were fed linoleic acid supplemented diets containing 20% (w/w) of partially hydrogenated low erucic acid rapeseed oil (HLRSO), partially hydrogenated herring oil (HHO), olive oil (OO) and trierucin + triolein (TE) for 10 weeks. An additional two groups were fed partially hydrogenated low erucic acid rapeseed oil and partially hydrogenated herring oil without linoleic acid supplementation (HLRSO- and HHO-, respectively). Substantial amounts of trans fatty acids were incorporated into liver microsomes (12.6% in group HLRSO) and platelets (7.0% in group HLRSO-). This incorporation was not dependent on the dietary linoleic acid level. Hepatic microsomal delta5 -desaturase activity was significantly increased after HLRSO feeding compared to 00 feeding. Delta6 -Desaturase activity did not vary in the linoleic acid supplemented groups. Both delta5 -and delta6 -desaturase activities were significantly increased in groups without linoleic acid supplementation. Docosenoic acid was incorporated into platelet phospholipids in contrast to liver microsomes. In the platelet, docosenoic acid seemed to have a special preference for phosphatidylserine. Very small amounts were incorporated into platelet phosphatidylinositol. Feeding diets HLRSO, HHO and 00 did not influence rat platelet cyclooxygenase or 12-lipoxygenase activity. Platelets from rats fed TE, however, produced significantly less 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) than platelets from rats fed OO. Feeding of HLRSO- and HHO- resulted in a significantly diminished production of the arachidonic acid metabolites 12-HETE, 12-hydroxy-5,8,10-heptadecatrienoic acid (HHT) and 6-keto-prostaglandin F1alpha in stimulated platelets and aorta. Thus, high dietary levels of trans isomers of monoenoic acids do not interfere with platelet cyclooxygenase or lipoxygenase activity provided sufficient amounts of linoleic acid are available.

The effect of dietary partially hydrogenated marine oils on desaturation of fatty acids in rat liver microsomes.

The influence of dietary partially hydrogenated marine oils on distribution of phospholipid fatty acids in rat liver microsomes was studied with particular reference to the metabolism of linoleic acid. Five groups of weanling rats were fed diets containing 20% (w/w) peanut oil (PO), partially hydrogenated peanut oil (HPO), partially hydrogenated Norwegian capelin oil (HCO), partially hydrogenated herring oil (HHO), and rapeseed oil (RSO) for 10 weeks. The partially hydrogenated oils were supplemented with linoleic acid corresponding to 4.6 cal % in the diets. Accumulation of linoleic acid and reduced amount of total linoleic acid metabolites were observed in liver microsomal phospholipids from rats fed partially hydrogenated oils as compared to PO feeding. The most striking effects on the distribution of omega 6-polyunsaturated fatty acids was obtained after feeding HHO, a marine oil with a moderate content of trans fatty acids in comparison with HPO but rich in isomers of eicosenoic and docosenoic acids. Liver microsomal delta 6- as well as delta 5-desaturase activities as measured in vitro were reduced in rats kept on HHO as compared to PO dietary treatment. The results obtained suggest that the dietary influence of partially hydrogenated marine oils on the metabolism of linoleic acid might be better related to the intake of isomeric eicosenoic and docosenoic acids than to the total intake of trans fatty acids.