BILATERAL EYE INJURY WITH BILATERAL BLOWOUT FRACTURE CAUSED BY A HIGH-PRESSURE WATER JET IN 16-YEARS-OLD FIREMAN GIRL. CASE REPORT.

PURPOSE: The aim is to present a report of a case of bilateral eye injury with bilateral blowout fracture caused by a high-pressure water jet from a fire hose in a 16-year-old girl during a competition for young firefighters. METHODS: We present a case report of a 16-year-old female patient with bilateral extensive eyelid contusion, oedema and lacerations of upper eyelids, lacerations of bulbar conjunctiva, contusion of both eyes, left optic nerve and bilateral blowout fractures of orbits.  Results: The initial ocular examination revealed "hand motion" in the right eye and no light perception in the left eye. Intraocular pressure was low in both eyes. Motility of both eyes was reduced, especially in the left eye. Lacerations of the eyelids and conjunctiva required a surgical procedure. No repair of the upper canaliculus of the left eye was attempted. Paralytic mydriasis and loss of lens accommodation in both eyes did not change throughout the follow-up period. A surgical procedure was applied to resolve the left blowout fracture, ocular motility of the right eye improved spontaneously. Traumatic cataract developed in the left eye within 1 year after injury, requiring cataract surgery and posterior capsule lens implantation, the right lens remained clear. Massive scarring of the retina and choroid in the lower parts of the eyes and in the macula reduced best corrected visual acuity in the right eye to 0.15 and in the left eye to 0.08. Intraocular pressure is within normal limits without any glaucoma therapy. The follow up period is three years. CONCLUSION: This is the first reported case of child with an eye injury caused by a high-pressure water jet during a sports activity.

Familiarization effects on the behavioral disinhibition of the cerebellar Lurcher mutant mice: use of the innovative Dual Maze.

Anxiety-related behaviors in mice are often assessed over short periods starting immediately after introducing the animals in a dedicated apparatus. In these usual conditions (5-10 min periods), the cerebellar Lurcher mutants showed disinhibited behaviors characterized by abnormally high exploration of the aversive areas in the elevated plus-maze test. We nevertheless observed that this disinhibition sharply weakened after 10 min. We therefore decided to further investigate the influence of the disinhibition on the intrinsic and anxiety-related exploratory behaviors in Lurcher mice, with a special focus on familiarization effects. To this end, we used an innovative apparatus, the Dual Maze, permitting to tune the familiarization level of animals to the experimental context before they are faced with more (open configuration of the device) or less (closed configuration of the device) aversive areas. Chlordiazepoxide administration in BALB/c mice in a preliminary experiment confirmed both the face and the predictive validity of our device as anxiety test and its ability to measure exploratory motivation. The results obtained with the Lurcher mice in the open configuration revealed that 20 min of familiarization to the experimental context abolished the behavioral abnormalities they exhibited when not familiarized with it. In addition, their exploratory motivation, as measured in the closed configuration, was comparable to that of their non-mutant littermates, whatever the level of familiarization applied. Exemplifying the interest of this innovative device, the results we obtained in the Lurcher mutants permitted to differentiate between the roles played by the cerebellum in exploratory motivation and stress-related behaviors.

SIMULTANEOUS BILATERAL ACUTE ANGLE-CLOSURE GLAUCOMA IN MILLER FISHER SYNDROME.

PURPOSE: To report a case of patient with Miller Fisher syndrome, complicated by simultaneous bilateral acute angle-closure glaucoma in her slightly (+1.5) hyperopic eyes. METHODS: We present a case report of a 71-year-old female patient presenting with total ophthalmoplegia, areflexia, ataxia and bilateral acute angle-closure glaucoma. RESULTS: The initial ocular examination revealed hand motion in the both eyes and oedematic corneas. Initial intraocular pressure was immeasurable high (measurment by Tonopen Avia). Measurement was possible after intravenous Mannitol 20 % infusion on both eyes as 54 and 56 mm Hg, respectively. Local medical therapy of pilocarpine, timolol, dorsolamide and dexamethasone improve intraocular pressure into normal limits within several hours. Prophylactic peripheral Nd-YAG laser iridotomy was performed on a both eyes two days later. Systemic treatment involved plasma exchange and rehabilitation program. Subsequent cataract surgery on both eyes with posterior capsule lens implantation improve the best corrected visual acuity on right eye from 0.5 to 1.0 and the left eye from 0.5 to 0.8, respectively. Intraocular pressure is within normal limits without any glaucoma therapy. Follow up period is three years. CONCLUSIONS: This is the second reported case of patient with Miller Fisher syndrome and simultaneous bilateral acute angle-closure glaucoma and the fifth reported case of Miller Fisher syndrome and acute angle-closure glaucoma. Treatment for both conditions made a very good recovery.

Comparison of P19-derived neuroprogenitor and naive cell survival after intracerebellar application into B6CBA mice.

Mouse embryonic carcinoma cells (P19 line) were studied for both their survival and developmental potential in the intact cerebellum of B6CBA mice. The P19 cells were cultured and labelled with green fluorescent protein using transfection. Cells were used for transplantation either in the undifferentiated stage or after 3 days of neurodifferentiation induced by retinoic acid. The intracerebellar application was performed in 43 mice: group A (N = 21) received neuroprogenitors and group B (N = 22) received undifferentiated cells. The morphology of transplanted cells within the context of the surrounding cerebellar tissue was evaluated after 3 weeks. Naive P19 cells engrafted and survived in the cerebellum of 7 of the 22 adult mice (survival rate 31.8 %). Neuroprogenitors survived in 13 of the 21 mice (survival rate was 61.9 %). Since the cut-off is P < 0.05, the difference is not statistically significant (P = 0.069). An expansive appearance of the graft was significantly more frequent (P = 0.0047) in naive P19 cells than in neuroprogenitors. In mice in which the grafts did not survive, no marks of grafted cells or only fluorescing detritus were found. In conclusion, this is the first study to track the fate and morphology of embryonic carcinoma cells transplanted into the cerebellum, confirming that neuroprogenitors derived from embryonic carcinoma cells can settle in the host tissue and differentiate according to the surrounding conditions. With further validation, the embryonic carcinoma cells could become a valuable model with which to study the impact of cell therapy on neurodegenerative diseases.

Characterization of P19 cells during retinoic acid induced differentiation.

The aim of our study was to characterize mouse embryonal carcinoma (EC) cells P19 in different stages of retinoic acid induced neurodifferentiation by two methods, immunocytochemistry and RT qPCR. The characterization of the cells is crucial before any transplantation into any model, e.g. in our case into the mouse brain with the aim to treat a neurodegenerative disease. Specific protein markers (MAP-2, OCT-4, FORSE-1) were detected by immunocytochemistry in the cell cultures. The mRNA expression levels of PAX-6, MASH-1, Brachyury, GATA-4 and AFP were determined by RT qPCR method. HPRT was used as a housekeeping gene. The degree of differentiation can be characterized by expression of analyzed genes. The presence of OCT-4 and FORSE-1 proteins in undifferentiated pluripotent cells and the presence of dendrite specific MAP-2 in neuroprogenitors was detected. The expression levels of PAX-6 and MASH-1 increased and expression of Brachyury decreased during the neurodifferentiation process. The expression levels of GATA-4 and AFP were the highest after induction of differentiation with retinoic acid. Detailed characterization of cells before transplantation experiments can contribute to better understanding of their effect.

Glutamate receptor block in Lurcher mutant mice during ontogeny and its effect on hippocampal long-term potentiation.

Basic evaluation of the effect of chronic NMDA glutamate receptor (NMDAR) blockade on the hippocampal long-term potentiation (LTP) was performed in an animal model of inborn olivo-cerebellar degeneration (Lurcher mutant mice, LMM). NMDA receptor antagonist MK-801 was administered to mice in the dose 0.2 mg/kg of body weight, daily during two periods of their ontogeny: D5-D26 and D91-D111. In the consecutive 15 days some behavioral characteristics were studied using special methods for physical activity testing. Then LTP was investigated in LMM and also in their healthy littermates which served as controls (wild-type, WT). LTP in animals pre-treated with MK-801 showed significant long-term suppression of NMDAR activity, in both WT and LMM despite certain small differences between them. Our results show that cerebellar pathology on one hand and a physical activity on the other hand can influence the LTP in hippocampal region. It can be concluded that the results support the ideas of close functional cooperation between the brain structures which are involved in mechanisms of learning and memory.

Changes of motor abilities during ontogenetic development in Lurcher mutant mice.

Lurcher mutant mice represent a natural model of olivocerebellar degeneration. This degeneration is caused by a mutation of the gene for the delta2 glutamate receptor. Lurcher mutants suffer from cerebellar ataxia and cognitive functions deficiency as a consequence of excitotoxic apoptosis of Purkinje cells in the cerebellar cortex and a secondary decrease of granule cells and inferior olive neurons. This process finishes by the 90th day of postnatal life, but already by 14 days, the Purkinje cells are damaged and the ataxia is fully developed. Purkinje cells die by apoptosis within the first 3 weeks of life. The aim of our work was to study the development of motor functions in the course of the ontogenetic development in Lurcher mutant mice of the B6CBA strain and to compare it with wild type mice of the same strain. Mice aged 2, 3, 6, 9, and 22 weeks were used in our experiment. Motor skills were examined using four standard tests: the horizontal wire, rotating cylinder, footbridge and slanting ladder. Our findings in Lurcher mutant mice show a significant increase of motor abilities up to the sixth postnatal week and selective decrease early after this period. This improvement of motor skills is caused by the physiological development of musculature and the nervous system, probably with some contribution of plasticity of the maturing brain. The cause of the decline of these abilities immediately after the completion of the development is unknown.

Hippocampal long-term potentiation in adult Lurcher mutant mice: effect of embryonic cerebellar graft and motor training.

Possible effect of trophic factors from embryonic cerebellar graft transplanted in adult Lurcher mutant mice on LTP as electrophysiological marker of learning and memory process was studied. Also the combination of the transplantation and long-term forced motor training was investigated. An evaluation of LTP ability in four animal groups (transplanted, sham-operated, with and without forced motor activity) and comparison among them showed the highest LTP improvement in the group with combination of both influences (ie. transplantation and motor training).

The time-dependent block of NMDA glutamate receptor influences hippocampal LTP in inborn cerebellar degeneration mouse model.

The effect of single dose of NMDA glutamate receptor blockage administration on the hippocampal LTP was evaluated in animal model of inborn cerebellar degeneration. We compared the level of possible LTP blockade in two groups of animals, Lurcher mutant mice and their healthy littermates which served as controls. In the second part of the study we tested group of mice which were influenced repeatedly by the same NMDA blocker (MK-801) during behavioral experiments. Our results suggest a similar effect of blockade either after single or chronic MK-801 administration; both of them practically disrupted LTP generation with differences between healthy and neurodegenerative animals.

Motor and visuospatial abilities in a model of olivocerebellar and retinal degeneration--Lurcher mutant mice of C3H strain.

Lurcher mutant mice represent a natural model of olivocerebellar degeneration. They suffer from loss of Purkinje cells and decreased number of granule cells and inferior olive neurons. The degeneration leads to cerebellar ataxia and deterioration of cognitive functions. Some animals of the C3H strain have also the retinal degeneration. The aim of the study was to analyze the morphology of cerebellar and retinal degeneration and to evaluate the ability of motor coordination and visuospatial orientation in C3H Lurcher mutant mice. Cerebella of Lurcher mutant and wild type mice were examined with several histological, histochemical and immunohistochemical methods. Motor coordination was tested on a bar, ladder and rotarod. Spatial orientation and learning were tested in the Morris water maze with visible or hidden platform. Histological examinations showed decreased numbers of Purkinje cell in Lurchers. Various histological methods brought different information about the course or stage of the cerebellar degeneration. Retinal degeneration was identified with hematoxyline-eosine staining very well. Lurchers performed worse in motor coordination tests and in both the spatial orientation and learning test. Retinal degeneration influenced negatively both the spatial learning and orientation. Motor tests were influenced by retinal degeneration only in the wild type mice. Wild type mice showed some ability of idiothetic navigation, which was not found in Lurchers.

Preliminary study of the effect of repeated motor training on spatial learning ability in adult lurcher mutant mice.

Lurcher mutant mice represent a model of olivocerebellar degeneration. They suffer from cerebellar ataxia and deterioration of cognitive functions. The aim of the work was to study the effect of repetitive enforced motor training on spatial learning ability and motor coordination in adult Lurcher mutant mice of the C57BI strain. Experimental mice were trained repetitively on a rotarod. Control mice were left without the training. Motor coordination was tested four times-before the training, in the third week of the training, at the end of the training and after a spatial learning test following the training. A rotarod of higher cylinder diameter and lower rotation speed was used. Spatial learning was examined using the Morris water maze. Trained animals achieved significantly better results than untrained mice in the 2nd and 3rd motor coordination test. In the last test following the spatial learning examination, untrained mice improved their performances so that there were no differences between trained and untrained group. In the Morris water maze trained mice showed higher spatial learning ability than untrained animals. Motor coordination capability of adult Lurcher mutant mice was improved by the training on rotarod but also by swimming during the experiment in the water maze. Repetitive motor activity led to increase of spatial learning ability.

Changes of dopamine receptors in mice with olivocerebellar degeneration.

Lurcher mutants are mice with functional mutation in the 82 glutamate receptor (GluRdelta2) that is predominantly expressed in cerebellar Purkinje cells and plays a crucial role in cerebellar functions. These mice display ataxia and impaired motor-related learning tasks. In order to elucidate the role of dopaminergic receptor system in coping with mutation in delta2 glutamate receptor the behavioral effect (spatial learning) of D1 dopamine receptor activation and inhibition and changes in D1-like and D2-like dopamine receptors in striatum, hippocampus and cerebellum in C57BI/7 and C3H Lurcher mutants and wild type mice were studied. We have found that Lurcher mutants were worse in the spatial learning but mice of both types reacted similarly to D1 dopamine receptor agonist (without effect) and antagonist (worsening). Moreover, Lurchers revealed substantial higher density of both D1-like and D2-like dopamine receptors in hippocampus in C57BI/7 strain, while in C3H strain only D1-like dopamine receptors were higher. In C57BI/7 strain, D-like dopamine receptors were lower in cerebellum; D2-like dopamine receptors were not affected. In the striatum, the receptor densities were similar to the wild type counterparts. Our results suggest specific participation of dopamine receptor system in coping with olivocerebellar degeneration.

Does transplantation of cerebellar embryonic tissue influence hippocampal LTP in adult Lurcher mutant mice?

Possible influence of embryonic cerebellar graft transplanted into the adult neurodegenerative brain in Lurcher mutant mice on long-term potentiation (LTP) in hippocampus was investigated. Evaluation of LTP ability and comparison with the tests of motor learning suggests similarities between magnitude of LTP and criteria of motor learning. Also interstrain differences were described. Our results support ideas about tight cooperation among brain structures which are involved in mechanisms of learning and memory.

Comparison of embryonic cerebellar graft survival in adult Lurcher mutant mice of strains C3H and C57Bl/7.

Lurcher mutant mice suffer from complete loss of cerebellar Purkinje cells. The aim of the work was to compare the solid embryonic cerebellar graft survival in adult Lurcher mutant mice derived from strains C3H and C57Bl/7 and to assess the morphology of the grafts. Embryonic cerebellar tissue was obtained from 12-13 days mice embryos expressing green fluorescent protein (GFP). Embryonic cerebellum was injected with a glass microcapillary into the cerebellum of adult Lurcher mutant mouse. Host mice were sacrificed 2-12 weeks after the transplantation. Brainstems and cerebella were examined histologically. The graft and graft derived GFP-positive cells were detected according to their green fluorescence. To visualise the structure of the graft Nissl staining was used. Graft survival percentage was evaluated in groups of mice sacrificed during the first, second or third month after the transplantation. The graft was found in all C57Bl/7 mice and in 90.9% of C3H mice examined within one month after the transplantation. In the second month the graft was present in 83.3% of C57Bl/7 and 50.0% of C3H mice. Till the third month the graft survived in 68.2% of C57Bl/7 mice and 22.2% of C3H mice. In C57Bl/7 mice a cerebellar structure was developed in the graft and migration of graft derived-cells to the host tissue was observed more often than in C3H mice. C567Bl/7 mice seem to be more suitable for experiments testing functional consequences of transplantation into the cerebellum requiring good long-term graft survival.

Neural functional and morphological consequences of retinal degeneration in C3H Lurcher mutant and wild type mice.

Lurcher mutant mice represent a model of olivocerebellar degeneration associated with the total functional elimination of the cerebellar cortex. The affected animals suffer from cerebellar ataxia and worsening of cognitive functions. Healthy littermates of Lurchers-wild type mice serve as controls. Except mentioned patterns some animals derived from the C3H strain exhibit signs of a hereditary retinal degeneration. The impact of the retinal degeneration on visuospatial abilities and on the neuronal morphology in visual projection of both C3H Lurcher mutant and wild type mice has been studied in this work. The Morrris water maze was used for examination of spatial learning when the animals learned to find a platform hidden under the water surface. Time of reaching the platform (escape latency) in individual experimental days as well as the swimming velocity was measured and the strategy of maze exploration was assessed. The presence of the retinal degeneration was proved histologically by means of classical hematoxillin-eosin method. The neurohistological examination of the superior colliculus and visual cortex was performed using a Ramón-Moliner modification of the Golgi method. The results obtained showed that retinal degeneration influenced the strategy of the maze exploration and caused generally worse results. The histological examination of eyes in animals with bad results confirmed presence of the retinal degeneration. The neurohistological examination of the brain visual projections of animals affected with the retinal defect showed most detectable changes in dendritic spines of the V1 cortex (lower density in general and less immature types).

Various methods of Purkinje cells transplantation and their functional response in Lurcher mutant mice.

Embryonic cerebellum was transplanted to adult Lurcher mutant mice affected with hereditary olivocerebellar degeneration and with resulting cerebellar ataxia. Grafts were applied as solid pieces of tissue or as cell suspensions. The aim was to replace Purkinje cells lost by the neurodegeneration with embryonic cells and to observe the effect on motor symptoms of cerebellar ataxia. Success rate of the two methods was also compared. Motor skills were tested before and in week intervals after the transplantation. The results were compared with sham-operated controls. When the solid graft was transplanted, the success rate was two times higher as compared with the cell suspension method. Fibre sprouting and cell migration from the graft to the host tissue was observed. Insignificant amelioration of motor skills was found in mice after the solid cerebellar tissue transplantation, while the cell suspension application had no effect.

Functional consequences of retinal degeneration in spatial orientation in C3H wild type and Lurcher mutant mice.

Lurcher mutant mice represent a model of genetically determined olivocerebellar degeneration. In the C3H strain there is also hereditary retinal degeneration. The aim of this work was to assess, whether the retinal degeneration influences spatial orientation and results of the spatial learning tasks. Two experiments in the Morris water maze were arranged. First, mice learned to find a platform position, which was linked to two labels on the periphery of the maze. In the second experiment the platform was removed and swimming velocity and preference of central or peripheral zone of the maze were assessed. Presence of the retinal degeneration was detected histologically. Both Lurcher mutant and wild type mice that exhibited long latencies in the first experiment were affected with the retinal degeneration, while animals that performed the trial well, had normal retina. Swimming velocity was not changed substantially. The maze exploration strategy was different in mice with and without the retinal degeneration.

Effect of whole-body exposure to high-frequency electromagnetic field on the brain electrogeny in neurodefective and healthy mice.

A direct registration of brain cortical and hippocampal activity during a high-frequency electromagnetic field (HF EMF) exposure was performed. All experimental procedures were done under urethane anaesthesia (20%, 2 g/kg i.p.) in Lurcher mutant mice, wild type (healthy littermates) were used as controls. Experimental animals were exposed to the HF EMF with frequency corresponding to cellular phones. Our method is based on the use of gel electrodes (silicon tubes or glass microcapillaries filled with agar) where the connection with classical electrodes is located out of HF EMF space. ECoG evaluation showed a distinct shift to lower frequency components but clear effect has been observed only in wild type (healthy) mice whereas in Lurcher mutant mice only gentle differences between frequency spectra were found. Measurement of hippocampal rhythmicity showed gentle changes with increase of higher frequencies (i.e. opposite effect than in cortex) and changes in theta oscillations registered from a dentate gyrus and CA1 area in both types of animals (healthy and mutant). These findings support the idea about possible influencing the central nervous system by HF EMF exposure and support also some recent results about possible health risks resulting from cellular phones use.