Polymeric nanocapsules as a technological alternative to reduce the toxicity caused by meloxicam in mice.

This study determined whether meloxicam in nanocapsules modifies stomach and liver damage caused by free meloxicam in mice. Male Swiss mice were treated with blank nanocapsules or meloxicam in nanocapsules or free meloxicam (10 mg/kg, intragastrically, daily for five days). On the seventh day, blood was collected to determine biochemical markers (glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, total bilirubin, unconjugated bilirubin, albumin and alkaline phosphatase). Stomachs and livers were removed for histological analysis. There was no significant difference in the biochemical markers in the plasma of mice. Meloxicam in nanocapsules did not have an ulcerogenic potential in the stomach or cause lipid peroxidation in the stomach and liver. Free meloxicam increased the ulcerogenic potential in the stomach and lipid peroxidation in the stomach and liver. Meloxicam in nanocapsules caused less histological changes than free meloxicam. In conclusion, polymeric nanocapsules can represent a technological alternative to reduce the toxicity caused by meloxicam.

Anti-inflammatory effect of a selective 11ß-hydroxysteroid dehydrogenase type 1 inhibitor via the stimulation of heme oxygenase-1 in LPS-activated mice and J774.1 murine macrophages.

11ß-Hydroxysteroid dehydrogenase type 1 (11ß-HSD1) converts inactive cortisone to the active cortisol. 11ß-HSD1 may be involved in the resolution of inflammation. In the present study, we investigate the anti-inflammatory effects of 2-(3-benzoyl)-4-hydroxy-1,1-dioxo-2H-1,2-benzothiazine-2-yl-1-phenylethanone (KR-66344), a selective 11ß-HSD1 inhibitor, in lipopolysaccharide (LPS)-activated C57BL/6J mice and macrophages. LPS increased 11ß-HSD1 activity and expression in macrophages, which was inhibited by KR-66344. In addition, KR-66344 increased survival rate in LPS treated C57BL/6J mice. HO-1 mRNA expression level was increased by KR-66344, and this effect was reversed by the HO competitive inhibitor, ZnPP, in macrophages. Moreover, ZnPP reversed the suppression of ROS formation and cell death induced by KR-66344. ZnPP also suppressed animal survival rate in LPS plus KR-66344 treated C57BL/6J mice. In the spleen of LPS-treated mice, KR-66344 prevented cell death via suppression of inflammation, followed by inhibition of ROS, iNOS and COX-2 expression. Furthermore, LPS increased NFκB-p65 and MAPK phosphorylation, and these effects were abolished by pretreatment with KR-66344. Taken together, KR-66344 protects against LPS-induced animal death and spleen injury by inhibition of inflammation via induction of HO-1 and inhibition of 11ß-HSD1 activity. Thus, we concluded that the selective 11ß-HSD1 inhibitor may provide a novel strategy in the prevention/treatment of inflammatory disorders in patients.

Distinction of microsomal prostaglandin E synthase-1 (mPGES-1) inhibition from cyclooxygenase-2 inhibition in cells using a novel, selective mPGES-1 inhibitor.

Inflammation-induced microsomal prostaglandin E synthase-1 (mPGES-1) is the terminal enzyme that synthesizes prostaglandin E(2) (PGE(2)) downstream of cyclooxygenase-2 (COX-2). The efficacy of nonsteroidal anti-inflammatory drugs and COX-2 inhibitors in the treatment of the signs and symptoms of osteoarthritis, rheumatoid arthritis and inflammatory pain, largely attributed to the inhibition of PGE(2) synthesis, provides a rationale for exploring mPGES-1 inhibition as a potential novel therapy for these diseases. Toward this aim, we identified PF-9184 as a novel mPGES-1 inhibitor. PF-9184 potently inhibited recombinant human (rh) mPGES-1 (IC(50)=16.5+/-3.8nM), and had no effect against rhCOX-1 and rhCOX-2 (>6500-fold selectivity). In inflammation and clinically relevant biological systems, mPGES-1 expression, like COX-2 expression was induced in cell context- and time-dependent manner, consistent with the kinetics of PGE(2) synthesis. In rationally designed cell systems ideal for determining direct effects of the inhibitors on mPGES-1 function, but not its expression, PF-9184 inhibited PGE(2) synthesis (IC(50) in the range of 0.5-5 microM in serum-free cell and human whole blood cultures, respectively) while sparing the synthesis of 6-keto-PGF(1alpha) (PGF(1alpha)) and PGF(2alpha). In contrast, as expected, the selective COX-2 inhibitor, SC-236, inhibited PGE(2), PGF(1alpha) and PGF(2alpha) synthesis. This profile of mPGES-1 inhibition, distinct from COX-2 inhibition in cells, validates mPGES-1 as an attractive target for therapeutic intervention.

Surfactant protein A enhances mycobacterial killing by rat macrophages through a nitric oxide-dependent pathway.

Surfactant-associated protein A (SP-A) is involved in surfactant homeostasis and host defense in the lung. We have previously demonstrated that SP-A specifically binds to and enhances the ingestion of bacillus Calmette-Guerin (BCG) organisms by macrophages. In the current study, we investigated the effect of SP-A on the generation of inflammatory mediators induced by BCG and the subsequent fate of ingested BCG organisms. Rat macrophages were incubated with BCG in the presence and absence of SP-A. Noningested BCG organisms were removed, and the release of tumor necrosis factor-alpha (TNF-alpha) and nitric oxide were measured at varying times. TNF-alpha and nitric oxide production induced by BCG were enhanced by SP-A. In addition, SP-A enhanced the BCG-induced increase in the level of inducible nitric oxide synthase protein. Addition of antibodies directed against SPR210, a specific macrophage SP-A receptor, inhibited the SP-A-enhanced mediator production. BCG in the absence of SP-A showed increased growth over a 5-day period, whereas inclusion of SP-A dramatically inhibited BCG growth. Inhibition of nitric oxide production blocked BCG killing in the presence and absence of SP-A. These results demonstrate that ingestion of SP-A-BCG complexes by rat macrophages leads to production of inflammatory mediators and increased mycobacterial killing.

Influence of tolazoline on caudal epidural administration of xylazine in cattle.

Eight adult female cattle (6 Holstein, 1 Jersey, 1 Brown Swiss) were used to determine the antagonistic effects of tolazoline, and alpha 2-adrenoceptor antagonist, on xylazine-induced (via caudal epidural administration) depression of CNS, respiratory, and cardiovascular activity and rumen motility. A 2% solution of xylazine HCl was injected into the epidural space at the first coccygeal interspace, using a dosage of 0.05 mg/kg of body weight, diluted to a 5-ml volume with sterile water, and administered at a rate of approximately 1 ml/30 s. Eight minutes after xylazine injection, either tolazoline (0.3 mg/kg) or saline solution (4 ml) was administered IV. All 8 cattle were treated, using both regimens in a random sequence; at least 1 week elapsed between treatments. Epidurally administered xylazine induced caudal analgesia (S3 to coccyx), as evaluated by no response to superficial and deep muscular pinprick, and induced sedation, cardiopulmonary depression, and inhibition of rumen motility, but all cattle remained standing. Tolazoline effectively reversed xylazine-induced rumen hypomotility, and partially antagonized xylazine-induced cardiopulmonary depression without affecting sedation and desirable local (S3 to coccyx) analgesic effects.

Effects of idazoxan, tolazoline, and yohimbine on xylazine-induced respiratory changes and central nervous system depression in ewes.

We compared the ability of 3 alpha 2-adrenoreceptor antagonists, idazoxan (0.05 mg/kg), tolazoline (2 mg/kg), and yohimbine (0.2 mg/kg) to reverse xylazine (0.3 mg/kg)-induced respiratory changes and CNS depression in 6 ewes. Once weekly, each ewe was given a random IV treatment of xylazine, followed in 5 minutes by either an antagonist or 0.9% NaCl solution. Xylazine alone caused recumbency for 54.2 +/- 5.3 minutes (mean +/- SEM). Xylazine also increased respiratory rate and decreased PaCO2 for at least 45 minutes, but did not significantly change arterial pH or PaCO2. Idazoxan and tolazoline were equally effective in reversing the respiratory actions of xylazine; however, yohimbine was less effective in reducing the respiratory rate and was ineffective in antagonizing the decreased PaO2. Idazoxan and tolazoline decreased the duration of xylazine-induced recumbency to 6.3 +/- 0.6 and 9.5 +/- 2.3 minutes, respectively, whereas yohimbine did not significantly change this effect of xylazine. Thus, at the dosages studied, idazoxan and tolazoline appeared to be more effective than yohimbine in reversing the respiratory and CNS depressant actions of xylazine in sheep.

[The antagonism of ketamine/xylazine anesthesia ("Hellabrunn mixture") in wild zoo ruminants]. / Untersuchungen zur Antagonisierung der Ketamin/Xylazin-Anästhesie ("Hellabrunner Mischung") bei im Zoo gehaltenen Wildwiederkäuern.

The ability of the antagonists tolazoline, yohimbine and the combination of yohimbine with 4-aminopyridine to reverse the effects of the xylazine-component of the "Hellabrunn mixture" (125 mg/ml xylazine and 100 mg/ml ketamine) on nondomestic zoo ruminants is discussed. Arousal time, recovery time and changes in the parameter of circulatory and respiratory functions after antagonization are shown. Tolazoline is able to antagonize the xylazine effect completely within a short time. Using a dosage of 3-5 mg/kg there is a marked negative effect on the cardio-vascular system. Yohimbine in the used dosage of 0.25-0.3 mg/kg in non-domestic ruminants did not approve in its effects. Combining yohimbine (0.25-0.3 mg) with 4-aminopyridine (0.5 mg/kg) recovery time is about 30 minutes. The negative effect on the cardio-vascular system is less pronounced compared with tolazoline.

Antagonistic effect of idazoxan on xylazine-induced central nervous system depression and bradycardia in calves.

Two doses of an alpha 2-adrenoreceptor antagonist, idazoxan, were administered to reverse the CNS depressant and bradycardia effects of xylazine in calves. Once a week for 3 weeks, each of 6 calves were administered IV one treatment of: (1) 0.2 mg of xylazine/kg of body weight followed in 10 minutes by 1 ml of 0.9% NaCl, (2) 0.2 mg of xylazine/kg followed in 10 minutes by 10 micrograms of idazoxan/kg, or (3) 0.2 mg of xylazine/kg followed in 10 minutes by 30 micrograms of idazoxan/kg. The order of the 3 treatments in each calf was selected at random. Xylazine alone caused lateral recumbency for 27.2 +/- 3.0 minutes (mean +/- SEM). Idazoxan administered at dosages of 10 and 30 micrograms/kg shortened xylazine-induced lateral recumbency to 11.5 +/- 0.8 and 10.3 +/- 0.2 minutes, respectively. Calves given xylazine alone stood at greater than 60 minutes after the onset of recumbency. Idazoxan given at dosages of 10 and 30 micrograms/kg shortened the time to standing to 16.8 +/- 1.7 and 11.3 +/- 0.2 minutes, respectively. Idazoxan given at a dosage of 30 micrograms/kg also reversed xylazine-induced bradycardia. Results indicated that idazoxan should be a useful antidote for xylazine overdose in cattle.

Yohimbine reversal of ketamine-xylazine immobilization of raccoons (Procyon lotor).

Six adult raccoons (Procyon lotor) were sedated with a combination of ketamine hydrochloride (KH) at 10 mg/kg body weight and xylazine hydrochloride (XH) at 2 mg/kg body weight intramuscularly (i.m.). Twenty min after the KH-XH combination was given, yohimbine hydrochloride (YH) at either 0.1 mg/kg (Trial 1) or 0.2 mg/kg (Trial 2) body weight or a saline control (Trial 3) was administered intravenously (i.v.). The time to arousal, time to sternal recumbency and time to walking were recorded. These times were significantly shortened after YH administration [e.g., mean time to walking (MTW) at 0.2 mg/kg YH = 23.7 min] as compared to the saline controls (MTW = 108.8 min). Heart and respiratory rates both increased after YH administration, while body temperature remained constant. A fourth trial was performed using a higher ratio of KH to XH (45:1 rather than 5:1) to mimic sedation as performed in the field. The mean time to arousal (MTA) and MTW in this trial (1.3 and 23.7 min, respectively) were significantly shorter than controls and similar to YH trials performed after immobilization with 5:1 KH-XH. Yohimbine hydrochloride may be useful in field studies that require sedation of raccoons using KH-XH combinations.

Blockade of 5-hydroxytryptamine3 receptors prevents cisplatin-induced but not motion- or xylazine-induced emesis in the cat.

5-Hydroxytryptamine3 antagonists have been reported to prevent emesis elicited by cisplatin and radiation. This study investigated the possibility that drugs with this mechanism of action may be useful in preventing emesis elicited by other stimuli. The drugs ICS 205-930 (0.1 and 1.0 mg/kg) and MDL 72222 (0.1 and 1.0 mg/kg) were administered SC to cats before challenging them with either provocative motion or an emetic dose of xylazine. In no instance was a significant reduction in emesis evident. Zacopride was also administered before motion testing (0.01 to 10.0 mg/kg) and found to not have efficacy. To test the possibility that species or route of administration were factors in the negative results, 1.0 mg/kg of ICS 205-930 was administered SC before IV infusion of 7.5 mg/kg of cisplatin. There was a total suppression of emesis for the duration of the six-hour observation periods. This result verifies other work which found 5-hydroxytryptamine3 antagonists to be effective in preventing emesis elicited by cancer chemotherapeutic treatments. However, there is no evidence that they are effective in other syndromes, such as motion sickness and xylazine-induced emesis.

Antagonism of xylazine hydrochloride sedation in raptors by yohimbine hydrochloride.

The mean time to initial reversal response (MTIRR) and the mean time to perching (MTP) were measured in 34 raptors sedated with xylazine hydrochloride with dosages ranging from 1.0 to 20 mg/kg intravenously (i.v.) and 2.5 to 20.0 mg/kg intramuscularly (i.m.). Yohimbine hydrochloride, given i.v. (0.2 mg/kg), 30 min after the injection of the xylazine, shortened the MTIRR and MTP compared to the controls. No adverse effects were noted due to the use of yohimbine. Yohimbine appeared to be a safe and effective antagonist for xylazine sedation in raptors.

Neuroinhibition of xylazine induced emesis.

Xylazine produces retching and vomiting presumably by activation of the chemoreceptor trigger zone (CTZ). The purpose of this project was to investigate whether neuroinhibition can prevent xylazine vomiting. Inhibitory neurons in the cervical vagus nerve of cats were stimulated with implanted cuff electrodes. Female cats, weighing from 3 to 4 kg, were anaesthetized with pentobarbital for surgical implantation of electrodes. After full recovery from surgery, animals were tested in weekly sessions. Stimulation was via a pulse generator connected to photon coupled linear isolator supplying constant current. Videotape was used to record observations. The range of effective stimulation was 1-10 ma, 4-100 Hz and 0.3-0.6 msec. Stimulation was initiated thirty sec. after subcutaneous injection of xylazine, 0.66 mg/kg. Stimulation of the inhibitory nerve group of the cervical vagus was effective in preventing vomiting in over 85% of the experimental trials. In addition to preventing emesis during stimulation, the latency of xylazine emesis was increased over control values. Repeated experimental trials of stimulation coupled with xylazine injection could result in the complete absence of emesis.

Yohimbine increases plasma insulin concentrations and reverses xylazine-induced hypoinsulinemia in dogs.

Xylazine (1.1 mg/kg of body weight, IV), an alpha 2-adrenoreceptor agonist, suppressed the increase in plasma insulin concentration induced by glucose (0.6 g/kg, IV) in dogs. Yohimbine (0.11 mg/kg, IV), an alpha 2-adrenoreceptor antagonist, given 5 minutes after xylazine, reversed effects of xylazine, whereas yohimbine alone increased plasma insulin and decreased plasma glucose concentrations. Seemingly, alpha 2-adrenoreceptors exert a negative control of insulin release.

Effect of cocaine on xylazine analgesic activity.

The effect of cocaine on the antinociceptive action of xylazine (XLZ) was studied by the method of abdominal contortions in mice. While XLZ alone (0.5 mg/kg, sc) inhibited the contortions by 43%, cocaine (5 mg/kg, sc) produced 39% inhibition and combination of the two drugs reduced the contortions by 77%. Cocaine acted synergistically with XLZ, decreasing the ED50 from 0.42 to 0.13 mg/kg. The mechanism involved does not seem to be directly mediated by activation of alpha-2 receptors since pretreatment of the animals with yohimbine did not significantly influence the XLZ effect.

Immobilization of white-tailed deer by etorphine and xylazine and its antagonism by nalmefene and yohimbine.

White-tailed deer (Odocoileus virginianus) were immobilized with either 4.0 mg etorphine hydrochloride (ETOR) or 3.5 mg ETOR and 50.0 mg xylazine (XYL). Deer immobilized with ETOR only were given 4.0 mg nalmefene hydrochloride (NAL), a new opioid antagonist, 20 min after induction. Deer immobilized with ETOR and XYL received 3.5 mg NAL and 0.125 mg/kg yohimbine hydrochloride (YOH). The dose of 4.0 mg ETOR did not provide acceptable immobilization and was discontinued. A NAL:ETOR ratio of 1:1 was insufficient for complete and sustained antagonism of ETOR. Subsequently, deer were immobilized with ETOR and XYL as before which was then antagonized with 35.0 mg NAL and 0.125 mg/kg YOH. The 10:1 ratio of NAL:ETOR appeared to provide complete antagonism with no evidence of renarcotization. Although more study is required, NAL could become a useful antagonist for opioid-induced immobilizations.

Reversal of ketamine/xylazine anesthesia in the rabbit with yohimbine.

Ketamine and xylazine used in combination have been shown to be effective, easily administered, cost efficient agents for surgical anesthesia in the rabbit. The effect of xylazine on the central nervous system has been shown to be mediated through alpha-2 adrenergic receptors. Yohimbine, an alpha-2 adrenergic antagonist has been shown to reverse xylazine induced depression and partially antagonize ketamine in other species. We evaluated the antagonistic effect of yohimbine on ketamine/xylazine anesthesia in the rabbit. Six New Zealand White rabbits were anesthetized with intramuscular ketamine (50 mg/kg) and xylazine (10 mg/kg) to establish baseline parameters including respiratory rate, heart rate, and palpebral, pedal and postural reflex activity. Fourteen days later each rabbit was subjected to the same anesthetic regimen followed 30 minutes later by the intravenous administration of yohimbine (0.2 mg/kg). The duration of anesthesia estimated by the time elapsed between the loss and return of the palpebral reflex was reduced in the yohimbine treated trial (means = 29.7 +/- 1.9 minutes) compared to the control trial (means = 67.0 +/- 13.5 minutes). The palpebral reflex returned within 5 minutes following yohimbine treatment. Our results indicated that yohimbine is an effective antagonist of ketamine/xylazine anesthesia in the rabbit. Yohimbine decreases anesthetic duration after intravenous administration and also may aid in the control of undesirable anesthetic effects and overdosage.

Yohimbine hydrochloride reversal of ketamine hydrochloride and xylazine hydrochloride immobilization of Bengal tigers and effects on hematology and serum chemistries.

Six bengal tigers (Panthera tigris tigris) were immobilized five times at 2-wk intervals with ketamine hydrochloride (ketamine) and xylazine hydrochloride (xylazine) mixtures at different dose levels. Hematology and serum chemistry analyses on blood samples collected at each immobilization remained normal during the study. There were acute changes in hematocrit, chloride, potassium, glucose, and bilirubin as a function of xylazine dose level. The effect of yohimbine hydrochloride (yohimbine) on the depth and duration of immobilization was evaluated in a crossover design with every animal serving as its own control at each dose. Administration of yohimbine resulted in recovery of the animals within 4-8 min in contrast to greater than 60 min with no yohimbine treatment. There were no adverse effects noted with the yohimbine treatment and the tigers did not exhibit a relapse over the next 24 hr. Yohimbine at a dose of 5-15 mg per adult tiger provided effective reversal of 50-150 mg of xylazine per tiger.