Tomasz Zarnowski

The NMDA antagonist procyclidine, but not ifenprodil, enhances the protective efficacy of common antiepileptics against maximal electroshock-induced seizures in mice

Procyclidine (up to 20mg/kg i.p.) did not influence the electroconvulsive threshold per se, but when given in a dose of 10mg/kg, it potentiated the protective activity of carbamazepine, diphenylhydantoin, phenobarbital and valproate, and in a dose of 20 mg/kg, that of diazepam against maximal electroshock-induced convulsions in mice. Ifenprodil increased the threshold for electroconvulsions when applied at 20 and 40 mg/kg (i.p.), but surprisingly, when combined with all antiepileptics tested, it did not influence their anticonvulsant actions. The chimney test in mice revealed, that application of procyclidine at 10 mg/kg together with phenobarbital and valproate, and procyclidine at 20 mg/kg with diazepam resulted in motor impairment. However, when procyclidine was applied at 10 mg/kg together with carbamazepine or diphenylhydantoin, no motor impairment was noted. The combined treatment of procyclidine (10 mg/kg) with carbamazepine, diphenylhydantoin, phenobarbital or valproate, as well as procyclidine (20 mg/kg) with diazepam caused significant worsening of long-term memory. Finally, procyclidine did not alter the total plasma levels of carbamazepine, diazepam, diphenylhydantoin, phenobarbital and valproate. It may be concluded that not all agents interfering with NMDA receptor complex-mediated events lead to the potentiation of the anticonvulsant activity of antiepileptic drugs.

Alterations of kynurenic acid content in the retina in response to retinal ganglion cell damage.

The present study is the first to examine the modulation of retinal kynurenic acid (KYNA) content in response to N-methyl-D-aspartate (NMDA)-induced cell death in adult rat retinal ganglion cells (RGC). Adult Brown Norway rats were intravitreally injected with NMDA or PBS. Surviving RGC were retrogradely labeled with fluorogold and counted in wholemounts of retinas 2, 7 and 14 days after injection. Retinal KYNA content was measured by HPLC at the same time points. RGC numbers decreased significantly 2, 7 and 14 days after NMDA injection if compared to control retinas. KYNA concentration increased significantly two days after NMDA-injection. However, 7 and 14 days after injection retinal KYNA content was found markedly decreased in NMDA-treated eyes as compared to controls. It is conceivable that KYNA deficiency is causally related to the pathology of excitotoxic retinal diseases.

Presence of kynurenic acid and kynurenine aminotransferases in the inner retina

Kynurenine aminotransferases (KATs I and II) are pivotal to the synthesis of kynurenic acid (KYNA), the only known endogenous glutamate receptor antagonist and neuroprotectant. This study is the first to identify KYNA in the rat retina and to examine immunohistochemically the distribution of KAT isoforms. As determined by HPLC, KYNA concentration in the retina was 99.9 ± 24.6 pmol/g wet wt. Immunohisto- chemical experiments showed that both KATs were present in the retina. KAT I was preferentially localised on Müller cell endfeet while KAT II was expressed in cells within the ganglion cell layer. In conclusion, KYNA is present and synthesised in the inner retina. This may suggest a modulatory role in glutamate-mediated retinal neurotransmission.

Neuroprotective effects of tempol on retinal ganglion cells in a partial optic nerve crush rat model with and without iron load.

Iron overload can contribute to oxidative stress in many tissues. We studied the effects of pretreatment with iron dextran on RGC loss in a calibrated partial optic nerve crush (PONC) model in rats, along with the protection offered by tempol (4-hydroxy-2,2,6,6-tetramethylpiperidinyl-1-oxyl, a membrane-permeable superoxide dismutase mimetic and free-radical scavenger), in the same experimental paradigm. A total of 40 rats in 6 groups of 5-8 animals each underwent PONC in one eye and sham crush in the other. Animals were pretreated with a single iron dextran load 24 h prior to PONC, and treated with tempol 6 h before and then once daily after PONC. Control animals were treated with PBS. RGC were retrogradely labeled with a fluorescent marker; all data are expressed in percent of the RGC count in the respective sham-treated eye. Immunohistochemistry was performed to visualize 3-nitrotyrosine, a marker of nitroxidative stress. PONC without iron pretreatment resulted in the survival of only 31.4% of labeled RGC after 7 days. Even fewer RGC (12.7%) survived after PONC with iron pretreatment. However, tempol in doses of 20 mg/kg of body weight (BW) significantly attenuated this effect when given as described above; in the group without iron pretreatment the number of surviving RGC doubled from 31.4% to 62.1%. In the group with iron pretreatment the survival rate of RGC increased even more pronouncedly, from 12.7% without tempol to 46.2% with tempol. Tempol in doses of 1 mg/kg BW and 5 mg/kg BW showed no significant rescue of RGC. Immunostaining showed nitrotyrosine-positive RGCs in PONC but not in sham-treated eyes and an increase in positive cells after iron load. Tempol treatment reduced nitrotyrosine staining in both the iron and non-iron groups. Our results demonstrate that PONC results in significantly greater RGC damage when iron pretreatment is performed, and that the compound tempol may provide additional protection for RGC in cases of neuronal damage both with and without prior iron treatment.

Pharmacodynamic and pharmacokinetic interactions between common antiepileptic drugs and acetone, the chief anticonvulsant ketone body elevated in the ketogenic diet in mice

Purpose:  Acetone is the principal ketone body elevated in the ketogenic diet (KD), with demonstrated robust anticonvulsant properties across a variety of seizure tests and models of epilepsy. Because the majority of patients continue to receive antiepileptic drugs (AEDs) during KD treatment, interactions between acetone and AEDs may have important clinical implications. Therefore, we investigated whether acetone could affect the anticonvulsant activity and pharmacokinetic properties of several AEDs against maximal electroshock (MES)–induced seizures in mice.

Comparison of static automated perimetry and semi-automated kinetic perimetry in patients with bilateral visible optic nerve head drusen

Purpose:  Until now there has been no standardized, systemic approach to diagnostics in patients with optic nerve head drusen (ONHD). This study compares visual field (VF) results obtained with static automated perimetry (SAP) and semi‐automated kinetic perimetry (SKP) in patients with bilateral visible ONHD.

Content of Kynurenic Acid and Activity of Kynurenine Aminotransferases in Mammalian Eyes

The present study investigated the kynurenic acid (KYNA) contents and kynurenine aminotransferase (KAT I and II) activity in structures of the human, monkey, rabbit and bovine eye. KYNA levels were investigated with HPLC and detected fluorimetrically. The activity of KAT I and II was assayed as quantitative analysis of newly synthesized KYNA in vitro. Mean KYNA levels (±SD) in the human retina and vitreous body were 36.8 ± 7.6 and 33.1 ± 6.2 pmol/g wet tissue weight, respectively. In human eyes, KAT I activity in the vitreous body was 0.57 ± 0.28, that of KAT II was 2.56 ± 0.69. KAT I activity in the retina was 3.42 ± 1.17 and that of KAT II 10.75 ± 9.2. (KAT activity is expressed as KYNA synthesis in picomoles per gram wet tissue weight per hour.) The values of KYNA and KAT observed in other mammalian species tested were in the same range. In conclusion, KYNA and KAT enzymatic activity are present in the structures of human and other mammalian eyes.

Variability in Isopter Position and Fatigue during Semi-Automated Kinetic Perimetry

Background: Assessment of factors influencing response variability to repeat presentations of III4e stimuli and the fatigue effect during semi-automated kinetic perimetry (SKP). Design: Prospective case series; setting: university hospital. Participants: 58 patients with severe visual field loss: 21 with glaucoma, 18 with retinitis pigmentosa, and 19 with postchiasmal visual pathway lesions. Methods: Following initial testing with three isopters (I2e or V4e, I4e and III4e), presentations of the III4e stimulus were repeated four times during the same session along identical vectors. Main Outcome Measures: Variability in III4e-isopter position (scatter of kinetic threshold) and the difference of isopter area between the first and four subsequent sessions (fatigue effect) of SKP were analyzed by diagnosis, age, visual acuity and reaction time (RT). Results: The mean scatter of the kinetic threshold was 2.5 degrees (deg) in the glaucoma group, 1.5 deg in the group with retinitis pigmentosa, and 1.7 deg in the group of patients with postchiasmal lesions. The difference in the isopter area between a single examination and four times repeated examination was 656 square degrees (deg2) in the glaucoma group, 104 deg2 in the retinitis pigmentosa group and 227 deg2 in the group of patients with postchiasmal lesions. Post-hoc regression analysis revealed that the variability of isopter position increased as the RT increased. Conclusion: The variability of III4e-isopter positionand fatiguewere most pronounced among glaucoma patients. RT is the most important factor influencing the variability of responses and fatigue during SKP, thus we propose that it can be used as a reliability indicator of SKP.

Ontogenic changes of kynurenine aminotransferase I activity and its expression in the chicken retina

Kynurenine aminotransferases are key enzymes for the synthesis of kynurenic acid (KYNA), an endogenous glutamate receptor antagonist. The study described here examined ontogenic changes of kynurenine aminotransferase I (KAT I) activity and its expression in the chicken retina. KAT I activity measured on embryonic day 16 (E16) was significantly higher than at all other stages (E12, P0 and P7). Double labeling with antibodies against glutamine synthetase showed that on P7 KAT I was expressed in Müller cell endfeet and their processes in the inner retina. Since KAT I activity is high in the late embryonic stages, it is conceivable that it plays a neuromodulatory role in the retina during the late phase of embryogenesis.

Effects of topical bevacizumab application on early bleb failure after trabeculectomy: observational case series

Background The aim of this study was to evaluate the influence of topical bevacizumab on the formation and function of filtering blebs in eyes with early bleb failure after antiglaucoma surgery. Methods Of all patients who underwent mitomycin-augmented trabeculectomy for glaucoma in the Department of Ophthalmology at the Medical University in Lublin, Poland, between March 2009 and March 2010, a total of 21 eyes from 20 patients with injected filtration bleb 9.8 ± 4.7 days after surgery were included in this observational case series. All patients were treated with standard steroid therapy and topical bevacizumab 5 mg/mL five times a day for 20.9 ± 9.8 days. Patients were followed up every other day, and a full eye examination was performed 14, 30, 60, and 180 days after initiation of treatment. Blebs were evaluated for vascularity by slit-lamp examination with concomitant photographic documentation and intraocular pressure measurement. Results Elevated functional bleb with significantly reduced vascularity was present in 16 eyes, and was flat and nonfunctional in five eyes. Intraocular pressure in all eyes decreased from a mean of 26.6 ± 9.6 mmHg before surgery to 14.6 ± 7.7 mmHg and 15.8 ± 8.3 mmHg at 2 and 6 months after surgery, respectively. Filtration bleb leak was noted in three eyes while on treatment with bevacizumab. Conclusion Topical application of bevacizumab might favor functional bleb formation after trabeculectomy in eyes with a high risk of failure.