BACKGROUND: Methamphetamine (MA) is a highly abused psychostimulant across all age groups including pregnant women. Because developing brain is vulnerable by the action of drugs, or other noxious stimuli, the aim of our study was to examine the effect of early postnatal administration of MA alone or in combination with enriched environment (EE) and/or stress of separate housing, on the levels of serotonin (5HT) in the hippocampus of male rat pups at three stages of adolescence (postnatal day (PND) 28, 35 and 45). MA (5 mg/kg/ml) was administered subcutaneously (sc) to pups (direct administration), or via mothers' milk between PND1 and PND12 (indirect administration). Controls were exposed saline (SA). Pups were exposed to EE and/or to separation from the weaning till the end of the experiment. RESULTS: On PND 28, in sc-treated series, EE significantly increased the muted 5HT in SA pups after separation and restored the pronounced inhibition of 5HT by MA. No beneficial effect of EE was present in pups exposed to combination of MA and separation. 5HT development declined over time; EE, MA and separation had different effects on 5HT relative to adolescence stage. CONCLUSIONS: Present study shows that MA along with environment or housing affect 5HT levels, depending on both the age and the method of application (direct or indirect). These findings extend the knowledge on the effects of MA alone and in combination with different housing conditions on the developing brain and highlight the increased sensitivity to MA during the first few months after birth.
- Klíčová slova
- Adolescence, Enriched environment, Hippocampus, Methamphetamine, Serotonin,
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Chronic constriction injury (CCI) is widely used as an animal neuropathic pain model. Neuropathic pain is considered to exist when withdrawal latency to thermal stimulation is decreased after inducing a CCI to the sciatic nerve. However, it is known that CCI leads to changes in skin temperature and that skin temperature can affect withdrawal latency. Aim of this study was to compare withdrawal latencies of constricted and contralateral hind limbs, to thermal stimulation, at the same artificially-induced skin temperatures. METHODS: Neuropathic pain was induced by four ligatures on the left sciatic nerve in adult male Wistar rats. Withdrawal latencies were measured from the 11th to 14th day after ligation, in different ambient temperatures, using the plantar test (Hargreaves method). By changing ambient we produced different hind limb skin temperatures. RESULTS: Our results show that (1) CCI cause an increase in skin temperature; (2) the withdrawal latency was inversely related to ambient and skin temperature in the same manner for both the ligated and contralateral hind limbs; and (3) withdrawal latencies did not differ significantly for the ligated and contralateral hind limbs when the temperature of the hind limbs was artificially made the same (i.e., by changing the ambient temperature). CONCLUSIONS: Withdrawal latencies to thermal stimulation did not differ on ligated and contralateral hind limb after CCI to the sciatic nerve if the temperature of the hind limbs was artificially or mathematically made the same. This finding may have significant impact on the interpretation results of neuropathic pain research.
- Klíčová slova
- CCI, Pain, Plantar test, Temperature, Wistar rats,
- MeSH
- hyperalgezie etiologie MeSH
- konstrikce MeSH
- krysa rodu Rattus MeSH
- nervus ischiadicus MeSH
- neuralgie * MeSH
- potkani Wistar MeSH
- teplota kůže * MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Capillary electrophoretic separation of ketamine, norketamine, hydroxynorketamine, and dehydronorketamine was performed in the counter-current regime under the influence of oppositely-directed electroosmotic flow. For this purpose, the fused silica capillaries were covalently coated with the poly(acrylamide-co-3-acrylamidopropyl trimethylammonium chloride) copolymer (PAMAPTAC). The content of the cationic monomer APTAC in the polymerization mixture varied in the range 0-6 mol. % and the generated electroosmotic flow increased continuously in the 0-20 · 10-9 m2V-1s-1 interval. Importantly, it resulted in improved electrophoretic resolution of ketamine/norketamine, which increased from 0.8 for neutral PAM coating (i.e. 0% PAMAPTAC) to 3.0 for 6% PAMAPTAC. The determination of ketamine and its derivates in rat serum was performed in a 4% PAMAPTAC capillary with an inner diameter of 25 μm. The separation was performed in a 500 mM aqueous solution of acetic acid (pH 2.3). The clinical sample was deproteinized by the addition of acetonitrile to the serum and a large volume of the treated sample was injected directly into the capillary. The achieved limit of detection ranged from 2.2 ng/mL for dehydronorketamine to 4.1 ng/mL for hydroxynorketamine; the intra-day repeatability was 1.0-1.5% for the migration time and 2.8-3.3% for the peak area. The developed methodology was employed for time monitoring of ketamines in rat serum after intra venous administration of low doses of anaesthetic at a level of 2 μg per g of body weight.
- Klíčová slova
- Capillary electrophoresis, Clinical analysis, Coating, Contactless conductivity detection, Peak master, Stacking,
- Publikační typ
- časopisecké články MeSH
A method of capillary electrophoresis with contactless conductivity detection has been developed for non-enantioselective monitoring the anaesthetic ketamine and its main metabolite norketamine. The separation is performed in a 15 μm capillary with an overall length of 31.5 cm and length to detector of 18 cm; inner surface of the capillary is covered with a commercial coating solution to reduce the electroosmotic flow. In an optimised background electrolyte with composition 2 M acetic acid + 1% v/v coating solution under application of a high voltage of 30 kV, the migration time is 97.1 s for ketamine and 95.8 s for norketamine, with an electrophoretic resolution of 1.2. The attained detection limit was 83 ng/mL (0.3 μmol/L) for ketamine and 75 ng/mL (0.3 μmol/L) for norketamine; the number of theoretic plates for separation of an equimolar model mixture with a concentration of 2 μg/mL was 683 500 plates/m for ketamine and 695 400 plates/m for norketamine. Laboratory preparation of rat blood plasma is based on mixing 10 μL of plasma with 30 μL of acidified acetonitrile, followed by centrifugation. A pharmacokinetic study demonstrated an exponential decrease in the plasma concentration of ketamine after intravenous application and much slower kinetics for intraperitoneal application.
- Klíčová slova
- acetonitrile stacking, capillary electrophoresis, contactless conductivity detection, ketamine, pharmacokinetics,
- MeSH
- anestetika krev farmakokinetika MeSH
- elektrická vodivost MeSH
- ketamin analogy a deriváty krev metabolismus farmakokinetika MeSH
- krysa rodu Rattus MeSH
- limita detekce MeSH
- potkani Wistar MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- anestetika MeSH
- ketamin MeSH
- norketamine MeSH Prohlížeč
Poly(d,l-lactide)/polyethylene glycol (PLA/PEG) micro/nanofibers loaded with paclitaxel (PTX, 10 wt%) were prepared by needless electrospinning technology, which allows large scale production for real medicinal practice. The fiber structure and properties were investigated by several methods including scanning electron microscopy, nitrogen adsorption/desorption isotherm measurements, differential scanning calorimetry, and X-ray diffraction measurements to examine their morphology (fiber diameter distribution, specific surface area, and total pore volume), composition, drug-loading efficiency, and physical state. An HPLC-UV method was optimized and validated to quantify in vitro PTX release into PBS. The results showed that the addition of PEG into PLA fibers promoted the release of higher amounts of hydrophobic PTX over prolonged time periods compared to fibers without PEG. An in vitro cell assay demonstrated the biocompatibility of PLA/PEG fibrous materials and showed significant cytotoxicity of PTX-loaded PLA/PEG fibers against a human fibrosarcoma HT1080 cell line. The chick chorioallantoic membrane assay proved that PTX-loaded fibers exhibited antiangiogenic activity, with a pronounced effect in the case of the PEG-containing fibers. In vivo evaluation of PTX-loaded PLA/PEG fibers in a human fibrosarcoma recurrence model showed statistically significant inhibition in tumor incidence and growth after primary tumor resection compared to other treatment groups.
- Klíčová slova
- Antiangiogenesis, CAM assay, Local tumor recurrence, Needleless electrospinning, PLA/PEG micro/nanofibers, Paclitaxel quantification,
- MeSH
- buněčná smrt účinky léků MeSH
- difrakce rentgenového záření MeSH
- inhibitory angiogeneze farmakologie MeSH
- kur domácí MeSH
- lidé MeSH
- lokální recidiva nádoru patologie prevence a kontrola MeSH
- myši nahé MeSH
- nádorové buněčné linie MeSH
- nanovlákna chemie ultrastruktura MeSH
- nosiče léků chemie MeSH
- paclitaxel farmakologie MeSH
- polyestery chemie MeSH
- polyethylenglykoly chemie MeSH
- tělesná hmotnost MeSH
- teplota MeSH
- tumor burden účinky léků MeSH
- uvolňování léčiv * MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- inhibitory angiogeneze MeSH
- nosiče léků MeSH
- paclitaxel MeSH
- poly(lactide) MeSH Prohlížeč
- polyestery MeSH
- polyethylenglykoly MeSH
A sensitive capillary electrophoretic method with on-line sample preconcentration by large volume sample stacking has been developed for determination of the anti-microbial agent pentamidine. The separation is performed in a fused silica capillary coated with covalently bound hydroxypropyl cellulose, with an internal diameter of 50 μm and length of 31.5 cm; the background electrolyte was 100 mM acetic acid/Tris at pH 4.7. The stacking is tested using a model sample of 1 μM pentamidine dissolved in 25% infusion solution and 75% acidified acetonitrile. Stacking permits the injection of a sample zone with a length of 95% of the total capillary length to achieve an enhancing factor of 77 compared to low injection into 1.8% of the total capillary length, with simultaneous high separation efficiency of approximately 1 350 000 plates/m. Stacking is based on simultaneous application of a separation field and a hydrodynamic pressure to force the acetonitrile zone out of the capillary. This approach allows the determination of pentamidine in rat blood plasma using only 12.5 μL of plasma treated by the addition of acetonitrile in a ratio of 1:3 v/v. The attained LOD is 0.03 μM and the intra-day repeatability is 0.1% for the migration time and 1.0% for the peak area at the injection 28.3% of capillary length. The performed pharmacokinetic study with ten-second scanning of the blood reveals rapid dynamics of pentamidine in the arterial bloodstream, while the changes are much slower in the venous system.
- Klíčová slova
- Capillary electrophoresis, Clinical analysis, Large volume sample stacking, Pressure assisted separation,
- MeSH
- antiinfekční látky krev MeSH
- elektroforéza kapilární metody MeSH
- krysa rodu Rattus MeSH
- limita detekce MeSH
- lineární modely MeSH
- pentamidin krev MeSH
- potkani Wistar MeSH
- reprodukovatelnost výsledků MeSH
- tlak MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antiinfekční látky MeSH
- pentamidin MeSH
Glutamate carboxypeptidase II (GCPII), also known as prostate-specific membrane antigen (PSMA) or folate hydrolase, is a metallopeptidase expressed predominantly in the human brain and prostate. GCPII expression is considerably increased in prostate carcinoma, and the enzyme also participates in glutamate excitotoxicity in the brain. Therefore, GCPII represents an important diagnostic marker of prostate cancer progression and a putative target for the treatment of both prostate cancer and neuronal disorders associated with glutamate excitotoxicity. For the development of novel therapeutics, mouse models are widely used. However, although mouse GCPII activity has been characterized, a detailed comparison of the enzymatic activity and tissue distribution of the mouse and human GCPII orthologs remains lacking. In this study, we prepared extracellular mouse GCPII and compared it with human GCPII. We found that mouse GCPII possesses lower catalytic efficiency but similar substrate specificity compared with the human protein. Using a panel of GCPII inhibitors, we discovered that inhibition constants are generally similar for mouse and human GCPII. Furthermore, we observed highest expression of GCPII protein in the mouse kidney, brain, and salivary glands. Importantly, we did not detect GCPII in the mouse prostate. Our data suggest that the differences in enzymatic activity and inhibition profile are rather small; therefore, mouse GCPII can approximate human GCPII in drug development and testing. On the other hand, significant differences in GCPII tissue expression must be taken into account when developing novel GCPII-based anticancer and therapeutic methods, including targeted anticancer drug delivery systems, and when using mice as a model organism.
- Klíčová slova
- glutamate carboxypeptidase II, mouse animal model, neuronal disorders, prostate cancer, prostate‐specific membrane antigen,
- Publikační typ
- časopisecké články MeSH
Cachectic rheumatoid arthritis, the less frequent form of the disease, is associated with loss of fat mass and often more severe course of the disease. Its experimental model represents rat adjuvant arthritis (AA) characterized by edema, lack of appetite, sharp body weight and fat loss. As individual fat depots display functional differences, here we studied lipolytic activity and sensitivity to lipolytic stimuli of nodeless epididymal fat (eWAT) and perinodal mesenteric fat (mWAT) depots at the peak of AA. We also examined changes in catecholamine and cytokine levels involved in lipolysis in plasma and/or isolated adipocytes from both WATs to identify the contribution of local, adipocyte-based processes and/or systemic events to adiposity loss in cachectic rheumatoid arthritis. AA was induced to male Lewis rats by complete Freund's adjuvant. Groups of ad libitum-fed and pair-fed controls were used to distinguish the effects of food restriction from inflammation-induced cachexia. Adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and its phosphorylated form (pHSL) were analyzed by western blot. CRP and catecholamine levels in plasma or adipocyte lysates were determined using ELISA kits. Cytokine-induced neutrophil chemoattractant-1 (CINC-1/CXCL1), monocyte chemoattractant protein-1 (MCP-1/CCL2), IL-1β, IL-6, IL-10 and leptin in adipocyte lysate were analyzed by quantitative protein microarray. Plasma glycerol and FFA were measured spectrophotometrically. AA rats developed severe cachexia, with lower adiposity in mWAT compared to normal and pair-fed controls, whereas in eWAT the adiposity was similarly reduced in AA and pair-fed groups. ATGL levels in both WATs were not affected by AA or pair feeding. AA upregulated levels of HSL, pHSL and pHSL/HSL ratio in mWAT, whereas none of these parameters has changed in eWAT of AA rats or in either WATs of pair-fed rats. In AA rats plasma glycerol was elevated, whereas FFA concentration was reduced. Plasma norepinephrine and epinephrine were increased in AA compared with both groups of controls. In eWAT adipocytes, AA but not pair feeding, upregulated norepinephrine levels. In mWAT adipocytes, AA rats showed higher epinephrine levels than pair-fed controls. Leptin levels in both WATs were depleted in AA animals in accordance with body weight loss. None of the measured cytokines in eWAT and mWAT was enhanced. Our results demonstrate augmented lipolytic activity in mWAT and not eWAT during cachectic arthritis. The adipocyte-derived cytokines do not seem to contribute to activated lipolysis. We first demonstrated enhanced presence of norepinephrine in perinodal adipocytes that may contribute to the regulation of local lipolytic activity by auto/paracrine fashion and thus provide independent fuel supply to activated lymph nodes.
- Klíčová slova
- Adipocyte adrenaline, cachectic adjuvant arthritis, epididymal fat, lipolysis, mesenteric fat, rat,
- MeSH
- adrenalin biosyntéza MeSH
- artritida experimentální imunologie metabolismus MeSH
- biologické markery MeSH
- C-reaktivní protein MeSH
- epididymis metabolismus MeSH
- humorální imunita MeSH
- krysa rodu Rattus MeSH
- lipolýza MeSH
- mezenterium metabolismus MeSH
- modely nemocí na zvířatech MeSH
- sterolesterasa metabolismus MeSH
- tukové buňky metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- adrenalin MeSH
- biologické markery MeSH
- C-reaktivní protein MeSH
- sterolesterasa MeSH
While the effect of cortex stimulation on pain control is widely accepted, its physiological basis remains poorly understood. We chose an animal model of pain to study the influence of sensorimotor cortex stimulation on tooth pulp stimulation evoked potentials (TPEPs). Fifteen awake rats implanted with tooth pulp, cerebral cortex, and digastric muscle electrodes were divided into three groups, receiving 60 Hz, 40 Hz and no cortical stimulation, respectively. TPEPs were recorded before, one, three and five hours after continuous stimulation. We observed an inverse relationship between TPEP amplitude and latency with increasing tooth pulp stimulation. The amplitudes of the early components of TPEPs increased and their latency decreased with increasing tooth pulp stimulation intensity. Cortical stimulation decreased the amplitude of TPEPs; however, neither the latencies of TPEPs nor the jaw-opening reflex were changed after cortical stimulation. The decrease in amplitude of TPEPs after cortical stimulation may reflect its anti-nociceptive effect.
- MeSH
- biofyzikální jevy MeSH
- bolest patofyziologie MeSH
- časové faktory MeSH
- čelisti patofyziologie MeSH
- elektrická stimulace metody MeSH
- krysa rodu Rattus MeSH
- management bolesti * MeSH
- měření bolesti MeSH
- modely nemocí na zvířatech MeSH
- potkani Sprague-Dawley MeSH
- reflex fyziologie MeSH
- somatosenzorické evokované potenciály fyziologie MeSH
- somatosenzorické korové centrum fyziologie MeSH
- zubní dřeň inervace fyziologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Reactive oxygen species play an important role both in physiological and pathophysiological reactions. The aim of this study was to determine the role of free radicals and antioxidants in the development of visceral pain. Visceral pain was produced by colorectal distension (CRD) in adult rats. CRD was caused by insertion of a lubricated latex balloon into the descending colon and rectum followed by inflation to 80mm Hg for 10min. During CRD, visceral pain was rated on 0-3.5 point scale. Oxidative stress was determined indirectly by measurement of free radical scavenging enzymes (glutathione peroxidase (GPx) and superoxide dismutase (SOD)) in the blood, liver and brain. Following CRD we observed (1) all rats expressed signs of visceral pain (overall rating was 1.83), (2) SOD and GPx levels were increased in the liver and blood, and decreased in the brain samples and (3) administration of the antioxidant Trolox, a water-soluble derivate of vitamin E, prior to CRD, prevented SOD and GPx changes in the liver, blood and brain, but did not affect pain scores. It was concluded, that CRD as a model of visceral pain, increases oxidative stress in animals, which could be prevented by prior administration of antioxidants; however, antioxidants did not attenuate signs of visceral pain caused by CRD.
- MeSH
- antioxidancia farmakologie MeSH
- bolest etiologie metabolismus prevence a kontrola MeSH
- chromany (dihydrobenzopyrany) farmakologie MeSH
- dilatace patologická komplikace MeSH
- glutathionperoxidasa krev metabolismus MeSH
- játra enzymologie MeSH
- kolon patologie MeSH
- krysa rodu Rattus MeSH
- mechanický stres MeSH
- měření bolesti MeSH
- mozek enzymologie MeSH
- oxidační stres * MeSH
- rektum patologie MeSH
- superoxiddismutasa krev metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid MeSH Prohlížeč
- antioxidancia MeSH
- chromany (dihydrobenzopyrany) MeSH
- glutathionperoxidasa MeSH
- superoxiddismutasa MeSH
