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There is evidence of the therapeutic potential of intranasal oxytocin for the treatment of pain and various psychiatric disorders, however, there is scant evidence that oxytocin reaches the brain. We quantified the concentration and distribution pattern of [125I]-radiolabeled oxytocin in the brains and peripheral tissues of rats after intranasal delivery using gamma counting and autoradiography, respectively. Radiolabel was detected in high concentrations in the trigeminal and olfactory nerves as well as in brain regions along their trajectories. Considerable concentrations were observed in the blood, however, relatively low levels of radiolabel were measured in peripheral tissues. The addition of a mucoadhesive did not enhance brain concentrations. These results provide support for intranasal OT reaching the brain via the olfactory and trigeminal neural pathways. These findings will inform the design and interpretation of clinical studies with intranasal oxytocin.

Objective Cerebral microdialysis (CMD) is a method used to measure the concentration of metabolites and glycerol in the interstitium of the brain. The aim of this study was to investigate the effect of parenterally applied medication and nutrition containing external free glycerol (EFG) on cerebral values of glycerol in patients monitored and treated for non-traumatic subarachnoid hemorrhage (SAH). Methods In 13 patients, the values of CG concentrations were measured using CMD. The amounts of parenterally applied EFG (in hourly intervals) were calculated from patient records. All data were gathered retrospectively. To analyze the association between the parameters of interest and their relationship, Spearman´s correlation and p-values were calculated. Results There was no evident relationship between the CG and EFG concentrations when the dataset was analyzed as a whole (r = -0.146). However, when the analysis was applied to single patients, a varying degree of correlations was discovered in 7 patients (r = 0.431-0.867). Conclusion The possible effect of externally administered glycerol contained in pharmaceuticals and nutrition on its brain concentrations must be considered when interpreting data of CMD (Tab. 2, Fig. 4,Ref. 16) Keywords: glycerol, microdialysis, brain, subarachnoid hemorrhage.

Target-controlled infusion systems are increasingly used to administer intravenous anaesthetic drugs to achieve a user-specified plasma or effect-site target concentration. While several studies have investigated the ability of the underlying pharmacokinetic-dynamic models to predict plasma concentrations, there are no data on their performance in predicting drug concentrations in the human brain. We assessed the predictive performance of the Marsh propofol model and Minto remifentanil model for plasma and brain tissue concentrations. Plasma samples were obtained during neurosurgery from 38 patients, and brain tissue samples from nine patients. Propofol and remifentanil concentrations were measured using gas chromatography mass spectrometry and liquid chromatography tandem mass spectrometry. Data were analysed from the nine patients in whom both plasma and brain samples were simultaneously obtained. For the Minto model (five patients), the median performance error was 72% for plasma and -14% for brain tissue concentration predictions. The model tended to underestimate plasma remifentanil concentrations, and to overestimate brain tissue remifentanil concentrations. For the Marsh model (five patients), the median prediction errors for plasma and brain tissue concentrations were 12% and 81%, respectively. However, when the data from all blood propofol assays (36 patients) were analysed, the median prediction error was 11%, with overprediction in 15 (42%) patients and underprediction in 21 (58%). These findings confirm earlier reports demonstrating inaccuracy for commonly used pharmacokinetic-dynamic models for plasma concentrations and extend these findings to the prediction of effect-site concentrations.

PURPOSE Fitting of MRS data plays an important role in the quantification of metabolite concentrations. Many different spectral fitting packages are used by the MRS community. A fitting challenge was set up to allow comparison of fitting methods on the basis of performance and robustness. METHODS Synthetic data were generated for 28 datasets. Short-echo time PRESS spectra were simulated using ideal pulses for the common metabolites at mostly near-normal brain concentrations. Macromolecular contributions were also included. Modulations of signal-to-noise ratio (SNR); lineshape type and width; concentrations of γ-aminobutyric acid, glutathione, and macromolecules; and inclusion of artifacts and lipid signals to mimic tumor spectra were included as challenges to be coped with. RESULTS Twenty-six submissions were evaluated. Visually, most fit packages performed well with mostly noise-like residuals. However, striking differences in fit performance were found with bias problems also evident for well-known packages. In addition, often error bounds were not appropriately estimated and deduced confidence limits misleading. Soft constraints as used in LCModel were found to substantially influence the fitting results and their dependence on SNR. CONCLUSIONS Substantial differences were found for accuracy and precision of fit results obtained by the multiple fit packages.

PURPOSE: Panobinostat, an orally bioavailable pan-HDAC inhibitor, has demonstrated potent activity in multiple malignancies, including pediatric brain tumors such as DIPG, with increased activity against H3K27M mutant cell lines. Given limited evidence regarding the CNS penetration of panobinostat, we sought to characterize its BBB penetration in a murine model. METHODS: Panobinostat 15 mg/kg was administered IV to 12 CD-1 female mice. At specified time points, mice were euthanized, blood samples were collected, and brains were removed. LC–MS was performed to quantify panobinostat concentrations. C(max) and AUC were estimated and correlated with previously published pharmacokinetic analyses and reports of IC-50 values in DIPG cell lines. RESULTS: Mean panobinostat plasma concentrations (ng/mL) were 27.3 ± 2.5 at 1 h, 7.56 ± 1.8 at 2 h, 1.48 ± 0.56 at 4 h, and 2.33 ± 1.18 at 7 h. Mean panobinostat brain concentrations (ng/g) were 60.5 ± 6.1 at 1 h, 42.9 ± 5.4 at 2 h, 33.2 ± 6.1 at 4 h, and 28.1 ± 4.3 at 7 h. Brain-to-plasma ratio at 1 h was 2.22 and the brain to plasma AUC ratio was 2.63. Based on the published human pharmacokinetic data, the anticipated C(max) in humans is expected to be significantly higher than the IC-50 identified in DIPG models. CONCLUSION: It is expected that panobinostat would be effective in CNS tumors where the IC-50 is in the low nanomolar range. Thus, our data demonstrate panobinostat crosses the BBB and achieves concentrations above the IC-50 for DIPG and other brain tumors and should be explored further for clinical efficacy.

The ability to formulate cubosomes and hexosomes with a single lipid by changing only the colloidal stabiliser presents a unique opportunity to directly compare the biological performance of these uniquely structured nanoparticles. This was explored here via the encapsulation and brain delivery of a model anti-seizure drug, phenytoin, in selachyl alcohol cubosomes and hexosomes. Nanoparticles were prepared with Pluronic® F127 or Tween 80® as the stabiliser and characterised. The internal nanostructure of nanoparticles shifted from hexosomes when using Pluronic® F127 as the stabiliser to cubosomes when using Tween 80® and was conserved following loading of phenytoin, with high encapsulation efficiencies (>97%) in both particle type. Cytotoxicity towards brain endothelial cells using the hCMEC/D3 line was comparable regardless of stabiliser type. Finally, in vivo brain delivery of phenytoin encapsulated in cubosomes and hexosomes after intravenous administration to rats was studied over a period of 60 min, showing cubosomes to be superior to hexosomes, both in terms of brain concentrations and brain to plasma ratio. While the role of stabiliser and/or internal nanostructure remains to be conclusively determined, this study is the first in vivo comparison of cubosomes and hexosomes for the delivery of a therapeutic drug molecule across the BBB and into the brain.

Idebenone (IDB) has demonstrated the potential to treat mitochondrial and neurodegenerative diseases, including Alzheimer's disease (AD). However, its therapeutic effects are compromised by poor compliance due to low bioavailability. The objective of this study is to fabricate IDB nanorods (IDBNRs) to improve oral bioavailability and increase concentrations in the brain in order to enhance therapeutic effects of IDB in the treatment of AD. IDBNRs showed desired sizes and rod-shaped morphologies. The release rate and the antioxidant activity of IDBNRs were improved relative to other delivery routes. The plasma and brain concentrations were enhanced due to rapid release into the systemic circulation. In behavioral tests, mice treated orally with IDBNRs showed amelioration of AD-induced impairment of learning and memory. Thus, because of improved efficiency of drug delivery, doses can be reduced, and the compliance and therapeutic experience of patients can be improved. IDBNRs may provide effective and convenient treatments for AD patients in the future.

Elasmobranch ecotoxicological investigations are complicated because accessing organs that accumulate organic contaminants is usually lethal. Several metrics among liver, muscle, and brain were evaluated to determine their relative organic contaminant loads and the efficacy of using muscle as a non-lethal proxy for liver. Liver contained the highest concentrations (368–4020 ng/g wet weigth [ww]) and greatest estimated total load of contaminants. Brain had higher toxin concentrations than muscle (4.18–84.2 ng/g ww versus 0.94–4.73 ng/g ww). Liver and brain were similar to each other in terms of contaminant detection occurrence and signature overlap, whereas muscle poorly reflected those of liver and brain. However, the identity of contaminants detected in muscle constituted those that substantially contributed to summed liver and brain concentrations. Thus, studies utilizing muscle as a non-lethal liver alternative to study organic contaminant exposure in elasmobranchs should craft questions with care, considering its limited ability to serve as an accurate proxy.

In vivo, real-time image-guided CED of a PET-imageable HDAC inhibitor is needed to improve survival in murine models of DIPG., Efforts at altering the dismal prognosis of pediatric midline gliomas focus on direct delivery strategies like convection-enhanced delivery (CED), where a cannula is implanted into tumor. Successful CED treatments require confirmation of tumor coverage, dosimetry, and longitudinal in vivo pharmacokinetic monitoring. These properties would be best determined clinically with image-guided dosimetry using theranostic agents. In this study, we combine CED with novel, molecular-grade positron emission tomography (PET) imaging and show how PETobinostat, a novel PET-imageable HDAC inhibitor, is effective against DIPG models. PET data reveal that CED has significant mouse-to-mouse variability; imaging is used to modulate CED infusions to maximize tumor saturation. The use of PET-guided CED results in survival prolongation in mouse models; imaging shows the need of CED to achieve high brain concentrations. This work demonstrates how personalized image-guided drug delivery may be useful in potentiating CED-based treatment algorithms and supports a foundation for clinical translation of PETobinostat.

Brain tumors are difficult to treat, in part, due to lack of adequate drug delivery across the blood–brain barrier (BBB). Although the BBB is compromised to some degree in brain tumors, it is not uniformly disrupted and some tumor locations have a relatively intact BBB. The presence of transporters (efflux) at the BBB limits the brain distribution of therapeutics intended for the treatment of CNS disorders. The differences in clearance processes, arising due to differences in the transporter expression and orientation in BBB vs BCSFB (blood–CSF barrier), make it complex to understand the specific influence of transporters on the CNS drug delivery, and also limit the use of CSF as a surrogate for unbound brain concentrations. Several opportunities to overcome the transporter-based barrier exist and it is important to recognize that solving the drug delivery problem will open many new doors for targeted therapies to be efficacious in the treatment of brain tumors.

Mercury (Hg) is a toxic element which has increased in marine environments for more than a century, due largely to anthropogenic activities, and biomagnifies in food chains to harmful levels in some top predators like waterfowl and seabirds. We analysed total mercury (THg) concentrations in blood, brain and muscle tissue from healthy specimens of 13 coastal and pelagic bird species from eastern and northern Canada to provide a baseline on current concentrations, especially for brain concentrations which are highly underrepresented in the literature. We also examined within and among tissues relationships of THg concentrations within individuals. THg concentrations were generally higher in pelagic species and scavenging gulls, when compared to coastal waterfowl. Brain and muscle tissue had similar concentrations of THg in the birds examined, but both of these tissues had lower concentrations that those found in blood. Our results, and that of a previous study, suggest that body condition has a large influence on blood THg concentrations and should be considered when using blood as a sampling medium. Many of the species we examined had tissue THg above levels known to cause deleterious, sublethal effects in some species.

Bumetanide is a loop diuretic that is proposed to possess a beneficial effect on disorders of the central nervous system, including neonatal seizures. Therefore, prediction of unbound bumetanide concentrations in the brain is relevant from a pharmacological prospective. A physiologically-based pharmacokinetic (PBPK) model was developed for the prediction of bumetanide disposition in plasma and brain in adult and paediatric populations. A compound file was built for bumetanide integrating physicochemical data and in vitro data. Bumetanide concentration profiles were simulated in both plasma and brain using the Simcyp PBPK model. Simulations of plasma bumetanide concentrations were compared against plasma levels published in the literature. The model performance was verified with data from adult studies before predictions in the paediatric population were undertaken. The adult and paediatric intravenous models predicted pharmacokinetic factors, namely area under the concentration-time curve, maximum concentration in plasma and time to maximum plasma concentration, within two-fold of observed values. However, predictions of plasma concentrations within the neonatal intravenous model did not produce a good fit with the observed values. The PBPK approach used in this study produced reasonable predictions of plasma concentrations of bumetanide, except in the critically ill neonatal population. This PBPK model requires more information regarding metabolic intrinsic clearance and transport parameters prior to further validation of drug disposition predictions in the neonatal population. Given the lack of information surrounding certain parameters in this special population, the model is not appropriately robust to support the recommendation of a suitable dose of bumetanide for use as an adjunct antiepileptic in neonates.

Exposure to high levels of manganese (Mn) in occupational settings is known to lead to adverse neurological effects. Since Mn is an essential nutrient, there are mechanisms that maintain its homeostatic control in the body, and there is some level of Mn in air that does not perturb Mn homeostasis. However, the Mn exposure concentrations at which no adverse effects are expected in occupational settings vary considerably across regulatory agencies. We set out to derive a Mn Occupational Exposure Level (OEL) for welders based on a review of studies that evaluated Mn exposure concentrations from welding fumes and: (1) neurological effects in welders; (2) levels of Mn in the brains of welders (via pallidal index [PI] estimated from magnetic resonance imaging [MRI]); (3) other biomarkers of Mn exposure in welders (i.e., blood and urine); and (4) Mn brain concentrations, PI, and corresponding neurological effects in non-human primates. Our analysis suggests uncertainty in quantifying dose-response associations for Mn from many of the occupational welding studies. The few welding studies that adequately estimate exposure suggest a possible OEL of 100-140μg/m3 for respirable Mn. This range is consistent with other epidemiology studies, studies of biomarkers of Mn exposure in welders, and with studies in non-human primates, though future studies could provide a stronger basis for deriving a Mn occupational guideline for welders.

Introduction Exposure to traumatic brain injury is a core risk factor that predisposes an individual to sporadic neurodegenerative diseases. We provide evidence that mechanical stress increases brain levels of hallmark proteins associated with neurodegeneration. Methods Wild-type mice were exposed to multiple regimens of repetitive mild traumatic brain injury, generating a range of combinations of impact energies, frequencies, and durations of exposure. Brain concentrations of amyloid β 1–42 (Aβ 1–42 ), total tau, and α-synuclein were measured by sandwich enzyme-linked immunosorbent assay. Results There was a highly significant main effect of impact energy, frequency, and duration of exposure on Aβ 1–42 , tau, and α-synuclein levels ( P 1–42 and tau ( P Discussion Dose-dependent and cumulative influence of repetitive mild traumatic brain injury–induced mechanical stress may trigger and/or accelerate neurodegeneration by pushing protein concentration over the disease threshold.

Studies in humans suggest that women progress more rapidly from initial cocaine use to addiction. Similarly, female rats can show more incentive motivation for cocaine than male rats do. Most preclinical studies on this issue have used self-administration procedures that provide continuous cocaine access during each session ("long-access" or LgA and "short-access"). However, intermittent access (IntA) cocaine self-administration better models the intermittency of human cocaine use. Here, we compared cocaine use in female and male rats that received ten, daily 6-hour LgA or IntA sessions. Cocaine intake was greatest under LgA, and female LgA rats escalated their intake. Only IntA rats (both sexes) developed locomotor sensitization to self-administered cocaine, and sensitization was greatest in females. Five and 25 days after the last self-administration session, we quantified responding for cocaine (0.083-0.75 mg/kg/infusion) under a progressive ratio (PR) schedule, a measure of motivation for drug. Across conditions, females earned more cocaine infusions than males under the PR schedule. Across sexes, IntA rats earned more infusions than LgA rats, even though IntA rats had previously taken much less cocaine. Cumulative cocaine intake significantly predicted responding for cocaine under the PR schedule in male LgA rats only. In IntA rats, the extent of locomotor sensitization significantly predicted responding under the PR schedule. Thus, LgA might be appropriate to study sex differences in cocaine intake, whereas IntA might be best suited to study sex differences in sensitization-related neuroadaptations involved in cocaine addiction. This has implications for modelling distinct features of cocaine addiction in preclinical studies.

Brain tissue is a useful supplement to blood in postmortem investigations, but reference concentrations are scarce for many opioids. Heroin cases may be difficult to distinguish from morphine cases as heroin and its metabolites are rapidly degraded. We present concentrations from brain and blood and brain-blood ratios of 98 cases where morphine was quantified. These cases were grouped according to the cause of death: A: The compound was solely assumed to have caused a fatal intoxication. B: The compound presumably contributed to a fatal outcome in combination with other drugs, alcohol or disease. C: The compound was not regarded to be related to the cause of death. The cases were further classified as heroin cases if 6-acetyl-morphine or noscapine were detected. The analyses were carried out using solid-phase extraction or protein precipitation followed by ultra high-performance liquid chromatography coupled to mass spectrometry. The average brain-blood ratios of morphine were 1.2 and 1.8 for 69 morphine and 29 heroin cases, respectively. Differences in the brain-blood ratios were found for cases where heroin and morphine were involved in the cause of death, either in combination or on its own (P

A whole-body permeability-rate-limited physiologically based pharmacokinetic (PBPK) model for cocaine was developed and adjusted with the pharmacokinetic data from studies with animals and reparametrized scaling to humans with the aim to predict the concentration-time profiles of the drug in blood and different tissues in humans. Estimated time course concentrations could be used as an interpretation tool by forensic toxicologists. The model estimations were compared successfully with pharmacokinetic parameters and time to peak for some effects reported in the literature. Once developed, the PBPK model was employed to predict the time course tissue concentrations reported in previous distribution studies introducing individualizing data. The heart and brain concentrations estimated by the model match adequately with the time and duration of some effects such as chronotropic and psychoactive effects, respectively. This work is the first attempt for employing PBPK modeling as a tool for forensic interpretation. Future modeling of other cocaine metabolite profiles or interaction when co-administered with other substances, such as alcohol, might be developed in the future.

This study explores dog plasma and brain fatty acid composition achieved after long-term supplementation at high DHA doses. A 90% concentrate of DHA Ethyl Ester (DHA-EE) administered by oral gavage to Beagle dogs at doses of 100, 500, 1000, and 2000 mg/kg bw/day for 8 weeks resulted in DHA increases in both plasma and brain. In a subsequent 9-month study, DHA-EE was administered at 150, 1000 and 2000 mg/kg bw/day. Plasma DHA increased between 150 and 1000 mg/kg bw/day but not between 1000 and 2000 mg/kg bw/day and there were increases from Day 1to 92 but not between days 92 and 273. Doses >500 mg/kg bw/day in the 8-week and all doses in the 9-month study resulted in DHA increases in the brain. The dose of 150 mg/k gbw/day is sufficient to achieve maximal brain concentrations if DHA is administered chronically. For shorter than 6 months of supplementation, higher doses are required.

A major limiting factor for the treatment of central nervous system (CNS) related disorders is the inability for drug substances to cross the blood-brain barrier. Some medications may possess dose-limiting systemic side effects that hinder their ability to reach maximum effective concentrations in the CNS. Over the last several decades, scientists have studied the ability for drugs to be transported from the nose directly to the brain, and compared to intravenous injections, many studies have reported higher brain concentrations from formulations administered intranasally. The primary focus of this paper is to review the formulation and device approaches that have been reported to increase drug delivery into the CNS through the nose-to-brain delivery pathway.

Background and Purpose The effects of ketamine in attenuating morphine tolerance have been suggested to result from a pharmacodynamic interaction. We studied whether ketamine might increase brain morphine concentrations in acute coadministration, in morphine tolerance and morphine withdrawal. Experimental Approach Morphine minipumps (6mg center dot day(-1)) induced tolerance during 5 days in Sprague-Dawley rats, after which s.c. ketamine (10mg center dot kg(-1)) was administered. Tail flick, hot plate and rotarod tests were used for behavioural testing. Serum levels and whole tissue brain and liver concentrations of morphine, morphine-3-glucuronide, ketamine and norketamine were measured using HPLC-tandem mass spectrometry. Key Results In morphine-naive rats, ketamine caused no antinociception whereas in morphine-tolerant rats there was significant antinociception (57% maximum possible effect in the tail flick test 90min after administration) lasting up to 150min. In the brain of morphine-tolerant ketamine-treated rats, the morphine, ketamine and norketamine concentrations were 2.1-, 1.4- and 3.4-fold, respectively, compared with the rats treated with morphine or ketamine only. In the liver of morphine-tolerant ketamine-treated rats, ketamine concentration was sixfold compared with morphine-naive rats. After a 2 day morphine withdrawal period, smaller but parallel concentration changes were observed. In acute coadministration, ketamine increased the brain morphine concentration by 20%, but no increase in ketamine concentrations or increased antinociception was observed. Conclusions and Implications The ability of ketamine to induce antinociception in rats made tolerant to morphine may also be due to increased brain concentrations of morphine, ketamine and norketamine. The relevance of these findings needs to be assessed in humans.

We disclose microemulsions (ME) of curcumin (CUR) with docosahexaenoic acid (DHA)-rich oil (CUR DHA ME) for targeted delivery to the brain. MEs of CUR (5 mg/mL) with and without DHA-rich oil (CUR Capmul ME) suitable for intravenous and intranasal administration exhibited negative zeta potential, globule size Cmax than CUR solution. Furthermore, high and sustained concentration was demonstrated even at 24 h, which was 8- and 2-fold higher than CUR solution and CUR Capmul ME, respectively. Brain concentrations following intranasal administration were, however, substantially higher as evident from higher Cmax and AUC and sustained compared to corresponding intravenous formulations signifying nose to brain targeting. The high brain concentration of CUR DHA ME is ascribed to the targeting efficiency enabled by DHA-mediated transport across the blood–brain barrier (BBB). Histopathological and nasal toxicity confirmed safety of the MEs. Concentration-dependent cytotoxicity in vitro, on human glioblastoma U-87MG cell line was observed with CUR DHA MEs and with the blank DHA ME, implying anticancer potential of DHA. The dramatically low IC50 value of CUR DHA ME (3.755 ± 0.24 ng/mL) is therefore attributed to the synergistic effect of CUR and DHA in the ME. The CUR concentration achieved with CUR DHA ME at 24 h which translated to >66-fold(intranasal) and >21–fold (intravenous) the IC50 value in the U-87MG cell line suggests great promise of CUR DHA ME for therapy of brain cancer by both routes.

Backgrounds: We studied the changes in blood-brain barrier (BBB) permeability and brain trace elements concentration in acute hypo-osmolality and pentylenetetrazol (PTZ)-induced seizures model in rats. Furthermore, the effect of acute hypo-osmolality on seizure activity (patterns) in the PTZ model, the correlation between seizure severity score (SS score) and changes of both of BBB permeability and brain trace elements concentration in rats were also studied. Materials and Methods: Four groups (18 animals /group) of adult male albino rats were included. The animal groups are: (1) Control group: received only a vehicle. (2) Hypo-osmolar group: received warm distilled water (DW). (3) Seizure group: injected with PTZ i.p. (4) Hypo-osmolar + seizure group: first injected with DW and then PTZ. During the next 30 minutes, the animals were observed for seizure activity. For evaluation of BBB permeability changes, serum S100s protein and Evan Blue (EB) dye content of brain tissue were evaluated. Additionally macroscopic and microscopic examinations of brain tissue were completed. Trace elements, magnesium (Mg), zinc (Zn), copper (Cu) and iron (Fe), concentration in brain tissue were estimated. Results: There were significant increase of EB content of brain tissue, and serum S100s in hypo-osmolar, seizure and hypo-osmolar + seizure groups, as compared with control animals. These parameters were significantly increased in the hypo-osmolar + seizure group when compared with either hypo-osmolar or seizure groups. These results were confirmed by macroscopic examination of brain tissue. Trace elements concentration of brain tissue, Mg, Zn, Cu and Fe, were significantly decreased in hypo-osmolar, seizure and hypo-osmolar + seizure groups in comparison with control animals. Histological examination of sections from the hilar region of dentate gyrus of the hippocampus revealed degenerative changes in the three groups of animals in comparison with control animals. Morphometric analysis showed a significant decrease in the number of cells in the hilar region of dentate gyrus in the three groups of animals in comparison with controls. Seizure activity significantly increased in hypo-osmolar + seizure as compared to seizure group. The correlation analysis revealed that significant –ve correlations were found between SS score and trace elements concentration, while significant +ve correlations were observed between SS score, and each of EB content and serum S100s protein in seizure and hypo-osmolar + seizure groups. Conclusion: Our study demonstrated that, hypo-osmolality and PTZ-induced seizures could result in increased BBB permeability with a decrease of brain trace elements concentration. This could play a vital role in the pathophysiology of epileptic seizures. In addition, hypo-osmolality can enhance neuronal excitability and severity of seizures in the PTZ-induced model in rats . Key words: Hypo-osmolality, seizures, Blood brain barrier, Brain trace elements

The synthesis and preclinical characterization of two novel, brain penetrating P2X7 compounds will be described. Both compounds are shown to be high potency P2X7 antagonists in human, rat, and mouse cell lines and both were shown to have high brain concentrations and robust receptor occupancy in rat. Compound 7 is of particular interest as a probe compound for the preclinical assessment of P2X7 blockade in animal models of neuro-inflammation.

The only method to quantify free extracellular levels of drugs in the brain of living animals is microdialysis. However, quantitative microdialysis has been hampered by methodological issues for decades. The problems arise from the need to establish the in vivo recovery for appropriate quantitation. In dealing with these issues the "dynamic no-net-flux" (DNNF) method seemed to be the experimental method of choice. Major disadvantages were, however, the need for a very high degree of bioanalytical precision and accuracy and the need for a large number of animals. Moreover, today we know that the experimental data are not always straightforward. To improve robustness and practicality of quantitative microdialysis sampling we modified the ultraslow microdialysis approach. Ultraslow microdialysis uses very low microdialysis flow rates (

The anaesthetic interaction between hexobarbital and thiopental in different combinations were investigated in male rats by using the isobolografic method. Data on dose, serum or brain concentrations at an EEG-criterion induced with a threshold method were utilized. The criterion was defined by a burst suppression in the EEG of 1 sec. or more (the “silent second”). With rats more than 115 days of age a synergistic interaction (potentiation) was obtained when a low dose of hexobarbital and a high dose of thiopental were used. The relationship between serum and brain concentrations of hexobarbital and thiopental in the different tested combinations gave no evidence for a pharmacokinetic interaction. Thus the synergistic interaction was located in the brain and probably related to the mechanisms of action. The same tests performed on rats at an age of approximately 90 days gave not only a potentiation when a low dose of hexobarbital and a high dose of thiopental were used, but also a potentiation when a high dose of hexobarbital and a low dose of thiopental were used. These results indicate an age-related change in sensitivity in the CNS between 90 and 115 days of age. Further investigations of this age-related change showed that this change from a potentiation to an additive interaction occurred between the age of 85 and 96 days.

Vigabatrin is undoubtedly one of the most exciting anti-epilepsy drugs in use today. Many open and controlled clinical trials have confirmed that it is particularly effective in controlling partial epileptic seizures with or without secondary generalization. Vigabatrin acts to increase GABA levels in the presynaptic nerve terminal by inhibiting the activity of GABA-transaminase. There is no direct correlation between the blood or brain concentration of vigabatrin and its clinical effect, so monitoring vigabatrin levels is not predictive of patient response. However, it is possible to relate the activity of vigabatrin to levels of GABA in the brain, measured by nuclear magnetic resonance spectroscopy (NMRS). NMRS studies show that following administration of vigabatrin, brain concentrations of GABA rise to about 2-3 times their baseline values. This 'extra' GABA is held within the nerve terminal, and is only released during synaptic transmission. Although there appears to be a clear dose-response relationship up to 3 g/day, it is not well documented if higher doses result in proportionately higher brain GABA levels. This finding seems to support the results of clinical studies suggesting that the optimal dose of vigabatrin may be 3 g/day. There is also some evidence for a correlation between the concentration of GABA in the brain and the clinical outcome. Continuing investigations using NMRS aim to confirm these preliminary findings, and to determine the time course and extent of changes in brain GABA levels after vigabatrin administration.

The distribution of an intravenous injectable nimodipine nanosuspension with mean particle size of both 300 and 650 nm in mice was systemically investigated compared with that of a nimodipine ethanol formulation (Nimotop) and a nanosuspension coated with Tween-80. The results showed that the 650-nm nanoparticles provided significantly higher drug concentrations in the liver, spleen and lungs because of their capture by Kupffer cells in the mononuclear phagocyte system, but lower drug concentrations in the brain compared with Nimotop and smaller nanoparticles. These nanoparticles failed to give increased brain concentrations even when coated with Tween-80. The 300-nm nanoparticles could effectively increase drug concentrations in the brain and remarkably reduce drug concentrations in the liver, spleen and lungs, indicating that the nimodipine nanosuspension may be a promising formulation with no ethanol, but the particle size must be small.

As 20 - 30% of patients are refractory to the presently available antiepileptic drugs, there is a strong need for the development of newer antiepileptics.A question arises whether stiripentol fulfills the criteria for a promising newer antiepileptic drug.A literature search, including experimental and clinical studies, with no time limit was performed.Stiripentol represents a group of drugs indirectly enhancing GABA-ergic neurotransmission and it exerts a protective activity in some experimental seizure models, including maximal electroshock (MES)-, pentylenetetrazol-, bicuculline-, strychnine-, and cocaine-induced convulsions. The drug potently inhibits metabolism of other antiepileptic drugs, significantly elevating their plasma and brain concentrations. This requires dosage adjustments of the concomitant antiepileptic drugs used in combination. In the form of an add-on therapy, stiripentol has proved to be effective in partial and atypical absence epilepsies. Considering its particular efficacy against severe myoclonic seizures in infancy (Dravet's syndrome), stiripentol has been awarded an orphan drug status for an adjunctive therapy of this difficult-to-treat condition.

A recently published physiologically based pharmacokinetic (PBPK) model successfully accounted for steady-state tissue manganese (Mn) concentration seen with normal dietary intakes and for biphasic, whole-body time-course profiles observed with tracer (54Mn) dosing. In this present study, PBPK modeling was used to evaluate Mn kinetics and brain concentrations in rats exposed to Mn both in their diet and by inhalation. Three published studies were used: (1) rats fed on diets ranging from 2 to 100 ppm, (2) rats on 125 ppm in diet and exposed via inhalation at 0.0 to 3.00 mg Mn/m3 each day for 14 d, and (3) rats to 0.1 or 0.5 mg Mn/m3 for 6 h/d, 5 d/wk over a 90-d period. The original model structure with well-mixed and "deep" compartments for each tissue could not describe rapid increases in tissue concentrations and rapid declines seen in high concentration inhalation studies. A second structure was developed that included (1) saturable, high-affinity binding of Mn in all tissues and (2) asymmetric diffusion from blood into brain (i.e., transport into and out of specific brain regions such as the striatum was described with different diffusion constants). This second model was consistent with liver and striatum experimental data. Preferential increases in some brain regions were predicted for exposures above 0.2 mg/m3 and had a rapid (i.e., 1 or 2 wk) return to steady-state levels. Multi-dose-route PBPK models for Mn based on this alternative model structure can be readily scaled to evaluate tissue Mn kinetics in other species and for human populations. Once validated across test animals, these PBPK models will be useful in tissue-dose based risk assessment with manganese.

The plasma pharmacokinetics and brain uptake of the novel neuroprotective agent AM-36 (1-(2-(4-chlorophenyl)-2-hydroxy)ethyl-4-(3,5-bis-(1, 1dimethylethyl)-4-hydroxyphenyl) methylpiperazine) were assessed over 72 h following i.v. administration to male Sprague-Dawley rats. At nominal i.v. doses of 0.2, 1 and 3 mg kg−1, AM-36 exhibited an extremely large volume of distribution (18.2–24.6 L kg−1) and a long terminal elimination half-life, ranging from 25.2 to 37.7 h. Over this dose range, AM-36 exhibited linear pharmacokinetics, with no apparent change in clearance, volume of distribution or dose-normalised area under the plasma concentration-time curve. AM-36 was very highly bound to plasma proteins (> 99.6%); however, this did not appear to affect the ability of AM-36 to permeate the blood-brain barrier. Following a single i.v. dose of AM-36 at 3 mg kg−1 to rats, brain concentrations were detected for up to 72 h, and the brain-to-plasma ratios were high at all time points (ranging from 8.2 at 5 min post-dose to 0.9 at 72 h post-dose). The very high brain uptake of AM-36 supports previous in-vivo efficacy studies demonstrating the neuroprotective effects of this compound when administered to rats with middle cerebral artery occlusion.

There are two recognized methods of estimating the brain concentrations of IV anesthetic drugs: (i) use of pharmacokinetic/pharmacodynamic (PK/PD) modeling of drug effect, from arterial concentrations and electroencephalogram changes, and (ii) direct measurement of the uptake of drug in the brain, by simultaneously measuring arterial and jugular concentrations and cerebral blood flow (mass-balance method). These two methods have not been directly compared. Because an accurate estimate of the time taken for transfer of anesthetic drug from arterial blood to its effect-compartment in the brain is critical for accurate effect-compartment dosing in IV anesthesia, we compared the PK/PD and mass-balance methods for propofol, methohexital, and ketamine in a sheep model.After instrumentation with arterial and sagittal-sinus cannulae, electrocorticogram, and sagittal sinus Doppler flow measurement seven adult sheep were given a random sequence of short anesthetic infusions with methohexital, ketamine, and propofol. Multiple blood samples were taken for measurement of the time course of the drug concentrations, and the electrocorticogram processed (approximate entropy, for propofol and methohexital and percentage high frequency time, for ketamine) to numerically quantify drug effect.Using the PK/PD method the t(1/2)K(eo) was 2.0 +/- 0.4 min for ketamine, 2.7 +/- 1.1 min for propofol, and was significantly shorter (0.3 +/- 0.1 min) for methohexital. PK/PD and the mass-balance methods did not differ in the times to peak effect.Both methods of calculating the delay in transfer of drug from arterial blood to brain give similar values. Methohexital crosses into the brain much faster than either propofol or ketamine.

Captive, juvenile, ranch-bred, male mink (Neovison vison) were fed diets containing various concentrations of methyl-mercury (MeHg) and selenium (Se) for a period of 13 weeks and then sacrificed to determine total Hg levels in fur, blood, brain, liver and kidneys and total Se concentrations in brain tissue. As MeHg concentrations in the diet increased, concentrations of total Hg in the tissues also increased with the highest level occurring in the fur > liver = kidney > brain > blood. Concentrations of Hg in the fur were correlated (r(2) > 0.97) with liver, kidney, blood and brain concentrations. The addition of Se to the mink diet did not appear to affect most tissue concentrations of total Hg nor did it affect the partitioning of Hg between the liver:blood, kidney:blood and brain:blood; however, partitioning of Hg between fur and blood was apparently affected.

The distribution, clearance, and bioavailability of (2S,6S)-hydroxynorketamine has been studied in the Wistar rat. The plasma and brain tissue concentrations over time of (2S,6S)-hydroxynorketamine were determined after intravenous (20 mg/kg) and oral (20 mg/kg) administration of (2S,6S)-hydroxynorketamine (n = 3). After intravenous administration, the pharmacokinetic parameters were estimated using noncompartmental analysis and the half-life of drug elimination during the terminal phase (t 1/2) was 8.0 ± 4.0 h and the apparent volume of distribution (V d) was 7352 ± 736 mL/kg, clearance (Cl) was 704 ± 139 mL/h per kg, and the bioavailability was 46.3%. Significant concentrations of (2S,6S)-hydroxynorketamine were measured in brain tissues at 10 min after intravenous administration, ∼30 μg/mL per g tissue which decreased to 6 μg/mL per g tissue at 60 min. The plasma and brain concentrations of (2S,6S)-hydroxynorketamine were also determined after the intravenous administration of (S)-ketamine, where significant plasma and brain tissue concentrations of (2S,6S)-hydroxynorketamine were observed 10 min after administration. The (S)-ketamine metabolites (S)-norketamine, (S)-dehydronorketamine, (2S,6R)-hydroxynorketamine, (2S,5S)-hydroxynorketamine and (2S,4S)-hydroxynorketamine were also detected in both plasma and brain tissue. The enantioselectivity of the conversion of (S)-ketamine and (R)-ketamine to the respective (2,6)-hydroxynorketamine metabolites was also investigated over the first 60 min after intravenous administration. (S)-Ketamine produced significantly greater plasma and brain tissue concentrations of (2S,6S)-hydroxynorketamine relative to the (2R,6R)-hydroxynorketamine observed after the administration of (R)-ketamine. However, the relative brain tissue: plasma concentrations of the enantiomeric (2,6)-hydroxynorketamine metabolites were not significantly different indicating that the penetration of the metabolite is not enantioselective.

S-adenosylmethionine (SAM) is an over-the-counter supplement marketed at improving mood and overall mental health. Although well known for decades to be an antidepressant, very little data exists w...

We investigated whether prostaglandins (PGs), proinflammatory mediators implicated in excitatory activity, are involved in Shigella -related seizures. Pretreatment with S. dysenteriae sonicate (2LD 50 ) enhanced mice response to pentylenetetrazole-induced seizures, without increase of brain concentrations of PGE 2 , PGD 2 or PGF 2α . Preinjection of NS-398, an inhibitor of cyclooxygenase-2, before treatment with Shigella sonicate, had no effect on seizures. The anticonvulsive PGD 2 increased after injection of 8 LD 50 of Shigella sonicate, which did not enhance seizures (32 pg/mg vs 26 pg/ml, p = 0.0063). The findings indicate that PGs are not involved in the enhanced seizure response after exposure to Shigella . However, induction of PGD 2 may play an inhibitory role.

Chronic use of non-steroidal anti-inflammatory drugs may reduce the risk or delay the onset of Alzheimer's disease. To date, only limited information exists on the brain distribution of these drugs. The objective of this study was to determine the absolute brain delivery of ibuprofen by using constant in vivo infusion in rats. Ibuprofen was infused to steady-state concentrations both in plasma and brain tissue. Ibuprofen levels in plasma and brain tissue were measured by RP-HPLC after the plasma and the brain samples were purified by protein precipitation and solid phase extraction, respectively. Results indicate that both plasma and brain concentrations reached steady-state within 6 h, and that the brain to plasma ratio of ibuprofen was only 0.02. Thus, limited brain penetration prevents the possible use of ibuprofen in treating or preventing neurodegenerative disorders such as Alzheimer's disease.

The proposition that environmental agents, such as diet, aluminum, and viruses, are as important as genetic factors in the etiology of Alzheimer's disease (AD) was advanced by the authors at the Challenging Views of Alzheimer's Disease meeting held in Cincinnati on July 28 and 29, 2001. Diet, dietary fat, and to a lesser extent, total energy (caloric intake), were found to be significant risk factors for the development of AD in a dozen countries, while fish consumption was found to be a significant risk reduction factor. An acid-forming diet, such as one high in dietary fat or total energy, can lead to increased serum and brain concentrations of aluminum and transition metal ions, which are implicated in oxidative stress potentially leading to the neurological damage characteristic of AD. Many of the risk factors for AD, such as cholesterol and fat, and risk reduction factors, such as whole grain cereals and vegetables, are shared with ischemic heart disease. Aluminum may cause neurological damage and a number of studies have linked aluminum to an increased risk for developing AD. The evidence for viral agents playing a role in AD is the strong association between the presence of HSV1 in brain and carriage of an apoE-epsilon4 allele in the case of AD patients but not of controls; statistical analysis shows the association is causal. Diet, aluminum, and viral infections may increase the prevalence of AD by eliciting inflammation, which may cause the neurological damage that results in AD.