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To maintain its functional abilities, the mature brain obtains energy from glucose produced in carbohydrate metabolism. When carbohydrates are eliminated from the diet, the energy comes from the oxidation of fatty acids. In this metabolic state called ketosis, ketone bodies are formed: β-hydroxybutyric acid (bHb), acetone, and acetoacetate as alternative source of energy passing through the blood–brain barrier easily. The ketosis state can be achieved through various strategies like caloric restriction, supplementation with medium-chain triglycerides, intense physical training, or ketogenic diet (KD). Using KD, drug-resistant epilepsy has been successfully treated in children and adults. It can also exert neuroprotective influences in cases of brain damage, glioblastoma multiforme, and Alzheimer's or Parkinson's diseases. Although many possible mechanisms of KD activity have been proposed, newer hypotheses appear with the research progress, mostly characterizing the brain under pathological but not normal conditions. Since different pathological conditions may affect the mechanism of KD action differently, additional research on the normal brain appears reasonable. For this purpose, young adult rats were treated with 4-month-lasting KD. Then, MRI structural measurements, spectroscopy, and tractography were performed. The procedures revealed significant increases in the concentration of glutamine, glutamate, glutathione and NAA, accompanied by changes in the pattern of neuronal connections of the striatum and hippocampal formation. This implies a possible involvement of these structures in the functional changes occurring in the brain after KD application. Thus, the investigations on the normal brain add important details concerning mechanisms underlying KD effects without their possible modification by a pathological status.

Glioblastoma multiforme (GBM) is a particularly malignant primary brain tumor. Despite enormous advances in the surgical treatment of cancer, radio- and chemotherapy, the average survival of patients suffering from this cancer does not usually exceed several months. For obvious ethical reasons, the search and testing of the new drugs and therapies of GBM cannot be carried out on humans, and for this purpose, animal models of the disease are most often used. However, to assess the efficacy and safety of the therapy basing on these models, a deep knowledge of the pathological changes associated with tumor development in the animal brain is necessary. Therefore, as part of our study, the synchrotron radiation-based X-ray fluorescence microscopy was applied for multi-elemental micro-imaging of the rat brain in which glioblastoma develops. Elemental changes occurring in animals after the implantation of two human glioma cell lines as well as the cells taken directly from a patient suffering from GBM were compared. Both the extent and intensity of elemental changes strongly correlated with the regions of glioma growth. The obtained results showed that the observation of elemental anomalies accompanying tumor development within an animal’s brain might facilitate our understanding of the pathogenesis and progress of GBM and also determine potential biomarkers of its extension. The tumors appearing in a rat’s brain were characterized by an increased accumulation of Fe and Se, whilst the tissue directly surrounding the tumor presented a higher accumulation of Cu. Furthermore, the results of the study allow us to consider Se as a potential elemental marker of GBM progression.

The ketogenic diet (KD) is a type of diet in which the intake of fats is significantly increases at the cost of carbohydrates while maintaining an adequate amount of proteins. This kind of diet has been successfully used in clinical therapies of drug-resistant epilepsy, but there is still insufficient evidence on its safety when used in pregnancy. To assess KD effects on the course of gestation and fetal development, pregnant females were fed with: (i) KD during pregnancy and lactation periods (KD group), (ii) KD during pregnancy replaced with ND from the day 2 postpartum (KDND group) and (iii) normal diet alone (ND group). The body mass, ketone and glucose blood levels, and food intake were monitored. In brains of KD-fed females, FTIR biochemical analyses revealed increased concentrations of lipids and ketone groups-containing molecules. In offsrings of these females, significant reduction of the body mass and delays in neurological development were detected. However, replacement of KD with ND in these females at the begining of lactation period led to regainment of the body mass in their pups as early as on the postnatal day 14. Moreover, the vast majority of our neurological tests detected functional recovery up to the normal level. It could be concluded that the ketogenic diet undoubtedly affects the brain of pregnant females and impairs the somatic and neurological development of their offspring. However, early postnatal withdrawal of this diet may initiate compensatory processes and considerable functional restitution of the nervous system based on still unrecognized mechanisms.

Disturbances of the early stages of neurogenesis lead to irreversible changes in the structure of the mature brain and its functional impairment, including increased excitability, which may be the basis for drug-resistant epilepsy. The range of possible clinical symptoms is as wide as the different stages of disturbed neurogenesis may be. In this study, we used a quadruple model of brain dysplasia by comparing structural and functional disorders in animals whose neurogenesis was disturbed with a single dose of 1 Gy of gamma rays at one of the four stages of neurogenesis, i.e. on days 13, 15, 17 or 19 of prenatal development. When reached adulthood, the prenatally irradiated rats received EEG teletransmitter implantation. Thereafter, pilocarpine was administered and significant differences in susceptibility to seizure behavioral symptoms were detected depending on the degree of brain dysplasia. Before, during and after the seizures significant correlations were found between the density of parvalbumin-immunopositive neurons located in the cerebral cortex and the intensity of behavioral seizure symptoms or increases in the power of particular EEG bands. Neurons expressing calretinin or NPY showed also dysplasia-related increases without, however, correlations with parameters of seizure intensity. The results point to significant roles of parvalbumin-expressing interneurons, and also to expression of NPY - an endogenous anticonvulsant and neuroprotectant reducing susceptibility to seizures and supporting neuronal survival. This article is protected by copyright. All rights reserved.

The present work applied a data envelopment analysis (DEA) model to assess changes in the technical efficiency of timber production at a period of transition in forest management priorities. The study material consisted of data on timber sold by Polish State Forests (PSF) and on its forest management inputs in the years 1990–2019. During the period of economic transition in Poland (1990–2003) the technical efficiency of timber production on average amounted to 0.809 and was highly variable. In the free-market period, that efficiency was much higher (on average 0.939) and more stable. This improvement was achieved by substantial layoffs in the PSF, and steady rise in the share and standing volume of mature stands, which made it possible to increase timber production while adhering to sustainable management principles. Analysis of the various categories of inputs to timber production revealed that the greatest decline, in labor costs, was attributable to a fourfold reduction in the workforce, and was accompanied by a 2.4-fold increment in timber production. On the other hand, logging costs increased due to, among other causes, the pursuit of environmentally friendly but more expensive harvesting procedures and reductions in clearcutting, which entailed more dispersed harvesting operations.

Birdwatching is one of the most sustainable types of nature-based tourism and, at the same time, a form of recreation that is developing very dynamically. Birdwatching is attracting more and more people, not only professionals, but also amateurs from many countries. Birdwatching research is still relatively embryonic, especially when compared to nature tourism or wildlife tourism. Our main aim was to determine preferences and opinions of birdwatchers visiting the largest national park in Poland, in relation to their different levels of involvement. The data were collected in 2018 from a survey of a sample of 357 Polish and foreign birdwatchers. Results showed that birdwatcher respondents were predominantly male, middle-aged, and living in a large city. An important tool described in this article is a new scale that assesses the level of involvement of individual people engaged in birdwatching activity. This scale corresponds well with the individual characteristics of birdwatchers. Most birdwatchers defined their birdwatching activity as a permanent rather than a temporary hobby and therefore considered it to be more of a lifestyle than a hobby. Engagement in birdwatching activity increased with age and frequency of trips. The two most important reasons for birding were ‘to be close to nature’ and ‘fascination with birds’. It has been proven that the development of birdwatching in the future will require a developed infrastructure enabling interaction with the objects of observation.

Traumatic brain injury (TBI), meaning functional or structural brain damage which appear as a result of the application of the external physical force, constitutes the main cause of death and disability of individuals and a great socioeconomic problem. To search for the new therapeutic strategies for TBI, better knowledge about posttraumatic pathological changes occurring in the brain is necessary. Therefore in the present paper the Fourier transform infrared microspectroscopy and Raman microscopy were used to examine local and remote biochemical changes occurring in the rat brain as a result of focal cortex injury. The site of the injury and the dorsal part of the hippocampal formation together with the above situated cortex and white matter were the subject of the study. The topographic and quantitative biochemical analysis followed with the statistical study using principal component analysis showed significant biomolecular anomalies within the lesion site but not in the area of the dorsal hippocampal formation and in the above situated white matter and cortex. The observed intralesional anomalies included significantly decreased accumulation of lipids and their structural changes within the place of injury. Also the levels of compounds containing phosphate and carbonyl groups were lower within the lesion site comparing to the surrounding cortex. The opposite relation was, in turn, found for the bands characteristic to proteins and cholesterol/cholesterol esters.

The animal models of seizures and/or epilepsy are widely used to identify the pathomechanisms of the disease as well as to look for and test the new antiseizure therapies. The understanding of the mechanisms of action of new drugs and evaluation of their safety in animals require previous knowledge concerning the biomolecular anomalies characteristic for the particular model. Among different models of seizures, one of the most widely used is the kindling model that was also applied in our study. To examine the influence of multiple transauricular electroshocks on the biochemical composition of rat hippocampal formation, Fourier transform infrared (FT-IR) microspectrosopy was utilized. The chemical mapping of the main absorption bands and their ratios allowed us to detect significant anomalies in both the distribution and structure of main biomolecules for electrically stimulated rats. They included an increased relative content of proteins with β-sheet conformation (an increased ratio of the absorbance at the wavenumbers of 1635 and 1658 cm–1), a decreased level of cholesterol and/or its esters and compounds containing phosphate groups (a diminished intensity of the massif of 1360–1480 cm–1 and the band at 1240 cm–1), as well as increased accumulation of carbohydrates and the compounds containing carbonyl groups (increased intensity of the bands at 1080 and 1740 cm–1, respectively). The observed biomolecular abnormalities seem to be the consequence of lipid peroxidation promoted by reactive oxygen species as well as the mobilization of glucose that resulted from the increased demand to energy during postelectroshock seizures.

Climate change is an important issue that increasingly affects our lives. One of the proposals for mitigating climate change is fighting biodiversity loss, which can support climate mitigation and adaptation actions. In Poland, the possibility of excluding large tracts of forest areas from use is being considered. The discussed the exclusion of forest land from use will affect the timber supply and market, especially for broadleaved wood. The main purpose of this analysis is to present a timber supply forecast, with a particular focus on the possibility of obtaining broadleaved hardwood timber in Poland from forests managed by State Forests National Forest Holding under three scenarios that assume different criteria for selecting forest areas for protection. The work was divided into two main phases: (1) the analysis of historical sales volume of wood products and average sale prices of hardwood during the period 2011–2020; (2) the preparation of a forecast of the potential possibility of maintaining broadleaved hardwood production in the three decades between 2020 and 2049. In the forecast, it was assumed that about 2.7 million hectares of planted and production forests are excluded from use in order to implement the provisions of the 2030 Biodiversity Strategy. In Scenario “I”, the supply of merchantable broadleaved roundwood volume will be reduced to 14%–18% that of Scenario “0”. In Scenario “II”, 55% of the “0” scenario is harvested, and in Scenario “III”, 33%–37% of the “0” scenario merchantable broadleaved roundwood is harvested. The introduction of restrictions on timber harvesting as a result of Poland’s compliance with European Union requirements in the area of environmental protection will lead to a significant reduction in the supply of timber on the market. This may lead to a further increase in timber prices and an increase in the importance of large timber buyers at the expense of local buyers. The recommendations contained in the policy objectives that the EU sets for the states should be supported by a thorough analysis when selecting areas for strict protection.

The article presents the results of a questionnaire survey aimed at determining social preferences in relation to running as a form of active leisure time in forests. These preferences were established on the basis of surveys conducted via the Internet by means of social media services. The survey was carried out in 2016 and covered 346 people living in Poland, interested in running and jogging. It involved people over 18 years of age. As a research tool, an Internet questionnaire was used, which included questions concerning the attractiveness of the forest for practicing running, the frequency of running events in the forest, and factors motivating to take part in the running event. Statistical analysis was performed considering the influence of personal characteristics (gender, age, place of residence) and experience in running on the preferences of respondents with the use of the chi-square test. Men dominated among the respondents. The majority of respondents were people aged 35–44 living in towns. Over half of the respondents declared that they practiced athletic or recreational running for more than 4 years. The results show that the Polish forests are an attractive place for running. The preferences varied significantly, mainly in relation to gender, age, and experience in running. Age differentiated the views of respondents on the attractiveness of the forest as a place for running. Participating in forest running events was determined by gender, place of residence, and experience in running. Participants of such events are more frequently men, residing in towns. It was observed that the frequency of participation in running events increases with experience. The attractiveness of the running event was associated mainly with its general atmosphere and the characteristics of the running trail. However, the views were diversified according to gender, age, residence, and experience in running.

The main goal of this study was to evaluate the elemental changes occurring in the main rat organs (kidneys, spleen, heart, brain) as a result of polyethylene glycol-coated magnetic iron oxide nanoparticles (PEG-IONPs) administration. For this purpose, 24 animals were divided into four equinumerous groups, and the three of them were intravenously injected with PEG-IONPs dispersed in 15% solution of mannitol in dose of 0.03 mg of Fe per 1 kg of body weight. The organs were collected 2 h, 24 h and 7 days passing from NPs administration, respectively, for the 2H, 24H, and 7D experimental groups. The forth group of animals, namely control group, was injected with 1 mL of physiological saline solution. For the analysis of subtle elemental changes occurring in the organs after nanoparticles injection, highly sensitive method of total reflection X-ray fluorescence spectroscopy was used. Obtained results showed that administration of even such low doses of PEG-IONPs may lead to statistically significant changes in the accumulation of selected elements within kidneys and heart. Two hours and 7 days from NPs injection, the Fe level in kidneys was higher compared to that of control rats. Elevated levels of Cu, possibly associated with systemic action of ceruloplasmine enzyme, were found within kidneys in 24H and 7D groups, while in heart the similar observation was done only for 24H group. The levels of Ca and Zn increased in kidneys and heart during the first 2 h from the injection and were again elevated in these organs 7 days later. The abnormalities in Ca and Zn accumulations occurring exactly in the same manner may suggest that these elements may interplay either in the mechanisms responsible for the detoxification of the PEG-IONPs or pathological processes occurring as a result of their action.

Malformations of cortical development (MCD) are a common cause of intractable seizures in humans. Among these, focal cortical dysplasia (FCD) poses an outstanding challenge. There are several subtypes of FCD that show significant variation in pathology and clinical presentation. All types exhibit disturbed cortical cytoarchitecture and increased propensity for seizures. The etiology is likely heterogenous, with mutations, specifically in genes related to mammalian target of rapamycin (mTOR), identified in only a subset of cases. A more complex mechanism, in which underlying genetic background interacts with early, pre- or perinatal injury or stress, has been proposed. Here, we used a well-established animal model of developmental malformations similar to MCD, induced by prenatal gamma irradiation. Previously, a significant variation between times of treatment has been shown, resulting in distinct and lasting patterns of dysplasia and differentially altered seizure propensity. We set out to describe the molecular background of these patterns by performing microarray analyses of hippocampal samples obtained from adult rats previously irradiated at distinct time points during gestation: E13, E15, E17 or E19 as well as controls. The analysis was performed in three conditions: naive, during latent phase after pilocarpine-induced status epilepticus and after 21 days of transauricular electric shocks. A set of 22 transcripts, some with known functions related to brain development, epilepsy and reaction to injury, was found to be altered between these groups across all treatments. We discuss the functional implication of these molecular differences, in an attempt to provide broader temporal and developmental context. © 2018 Wiley Periodicals, Inc. Develop Neurobiol 78: 718-735, 2018.

BACKGROUND Malformations of cortical development, such as focal cortical dysplasia, are commonly associated with intractable epilepsy. Multiple animal models were created in attempts to recapitulate features of human malformations of cortical development. These manipulations give rise to various focal or diffuse anatomical abnormalities, accompanied by altered susceptibility to epileptic seizures. Both in humans and in models of dysplasia, the question of timing of the initiating insult is important. METHODS Here, we used a rat model of cerebral dysplasia elicited by prenatal irradiation at gestational days (E) 13, 15, 17, or 19. Previous results suggest these animals are characterized by different patterns of dysplasia as well as different reactivity to seizurogenic stimuli in several seizure models. Rats were implanted with telemetric electroencephalography (EEG) transmitters. We compared EEG data from freely moving animals with anatomical observations obtained with MRI, as well as Western blotting and immunohistochemistry. RESULTS We performed spectral analyses of EEG signal, revealing differential regulation of specific bands (including delta, theta, alpha-beta, gamma) in animals with different patterns of dysplasia. Relative contribution of low-frequency activity in delta band is the lowest in E15 and the highest in E19. Conversely, higher frequency bands, corresponding to alpha/beta and gamma components, are reduced in E15 versus E19. This is accompanied by altered expression of glial markers in the E19 group. CONCLUSION To our knowledge, this constitutes the first quantitative description of power spectral properties in this experimental model, providing insight into mechanisms underlying the anatomical and electrophysiological abnormalities associated with brain dysplasia of different types. Birth Defects Research, 2017.© 2017 Wiley Periodicals, Inc.

The main purpose of this study was the determination and comparison of anomalies in creatine (Cr) accumulation occurring within CA3 and DG areas of hippocampal formation as a result of two high-fat, carbohydrate-restricted ketogenic diets (KD) with different ketogenic ratio (KR). To reach this goal, Fourier transformed infrared microspectroscopy with synchrotron radiation source (SRFTIR microspectroscopy) was applied for chemical mapping of creatine absorption bands, occurring around 1304, 1398 and 2800 cm-1. The samples were taken from three groups of experimental animals: control group (N) fed with standard laboratory diet, KD1 and KD2 groups fed with high-fat diets with KR 5:1 and 9:1 respectively. Additionally, the possible influence on the phosphocreatine (PhCr, the high energetic form of creatine) content was evaluated by comparative analysis of chemical maps obtained for creatine and for compounds containing phosphate groups which manifest in the spectra at the wavenumbers of around 1240 and 1080 cm-1. Our results showed that KD2 strongly modifies the frequency of Cr inclusions in both analyzed hippocampal areas. Statistical analysis, performed with Mann-Whitney U test revealed increased accumulation of Cr within CA3 and DG areas of KD2 fed rats compared to both normal rats and KD1 experimental group. Moreover, KD2 diet may modify the frequency of PhCr deposits as well as the PhCr to Cr ratio.

Disorders of neurogenesis at early developmental stages lead to irreversible structural and functional impairments of the brain. As further their consequences, increases in brain excitability and the development of drug-resistant epilepsy can frequently be observed in clinical cases. Mechanisms underlying these phenomena can also be examined on animal models of brain dysplasia. This study was conducted on rats with four degrees of brain dysplasia following exposure to gamma radiation on days 13, 15, 17, or 19 of prenatal development. When reached adulthood, the rats received electroencephalographic (EEG) transmitter implantation. Thereafter, pilocarpine was administered, and significant differences in susceptibility to seizures were detected depending on the degree of brain dysplasia. Before, during, and after the seizures, EEG was recorded in free moving animals. Additionally, the intensity of seizure behavioral symptoms was assessed. Strong and moderate correlations were found between the intensity of seizure behavioral symptoms, the power of particular EEG bands, and volumes of dysplastic brains and their regions. The data drew particular attention to correlations between variations in EEG spectra and changes in the midbrain and pons volumes. The results point to possible significant roles of these regions in the observed changes of susceptibility to seizures. Consequently, the frequently used experimental model was considered here not only as representing cases of cortical dysplasia but also of generalized, diffuse dysplasia of the whole brain.

Glioblastoma multiforme (GBM) is a primary brain tumor with a very high degree of malignancy and is classified by WHO as a glioma IV. At present, the treatment of patients suffering from GBM is based on surgical resection of the tumor with maximal protection of surrounding tissues followed by radio- and pharmacological therapy using temozolomide as the most frequently recommended drug. This strategy, however, does not guarantee success and has devastating consequences. Testing of new substances or therapies having potential in the treatment of GBM as well as detection of their side effects cannot be done on humans. Animal models of the disease are usually used for these purposes, and one possibility is the implantation of human tumor cells into rodent brains. Such a solution was used in the present study the purpose of which was comparison of elemental anomalies appearing in the brain as a result of implantation of different glioblastoma cell lines. These were two commercially available cell lines (U87MG and T98G), as well as tumor cells taken directly from a patient diagnosed with GBM. Using total reflection X-ray fluorescence we determined the contents of P, S, K, Ca, Fe, Cu, Zn, and Se in implanted-left and intact-right brain hemispheres. The number of elemental anomalies registered for both hemispheres was positively correlated with the invasiveness of GBM cells and was the highest for animals subjected to U87MG cell implantation, which presented significant decrease of P, K, and Cu levels and an increase of Se concentration within the left hemisphere. The abnormality common for all three groups of animals subjected to glioma cell implantation was increased Fe level in the brain, which may result from higher blood supply or the presence of hemorrhaging regions. In the case of the intact hemisphere, elevated Fe concentration may also indicate higher neuronal activity caused by taking over some functions of the left hemisphere impaired as a result of tumor growth.

In this study novel d-mannitol coated maghemite nanoparticles (MIONPs) are presented in terms of their influence on elemental homeostasis of living organisms and for this purpose highly sensitive total reflection X-ray fluorescence was used. Because of the biological indifference of d-mannitol and presumed lower toxicity of maghemite, compared to the most commonly used magnetite in nanomedicine, such nanoparticles seem to be promising candidates for biomedical applications. The examined dose of MIONPs was comparable with one of the lowest doses used in medical diagnostics. However, it should be emphasized that the amount of iron injected in this form is still significant compared to its total content in organs, especially in kidneys or the heart, and may easily disrupt their elemental homeostasis. The aim of the present study was to evaluate the elemental changes occurring in selected rat organs after injecting a low dose of MIONPs. The results were compared with those obtained for previously examined PEG-coated nanoparticles with magnetite cores. In the light of our findings the elemental changes observed after exposure to MIONPs were less extensive than those following PEG-coated magnetite nanoparticle administration.

Iron oxide nanoparticles (IONPs) have biomedical and biotechnological applications in magnetic imaging, drug-delivery, magnetic separation and purification. The biocompatibility of such particles may be improved by covering them with coating. In presented paper the biochemical anomalies of liver and kidney occurring in animals exposed to d-mannitol-coated iron(III) oxide nanoparticles (M-IONPs) were examined with Fourier transform infrared (FTIR) microspectroscopy. The dose of IONPs used in the study was significantly lower than those used so far in other research. Liver and kidney tissue sections were analysed by chemical mapping of infrared absorption bands originating from proteins, lipids, compounds containing phosphate groups, cholesterol and cholesterol esters. Changes in content and/or structure of the selected biomolecules were evaluated by comparison of the results obtained for animals treated with M-IONPs with those from control group. Biochemical analysis of liver samples demonstrated a few M-IONPs induced anomalies in the organ, mostly concerning the relative content of the selected compounds. The biomolecular changes, following exposition to nanoparticles, were much more intense within the kidney tissue. Biochemical aberrations found in the organ samples indicated at increase of tissue density, anomalies in fatty acids structure as well as changes in relative content of lipids and proteins. The simultaneous accumulation of lipids, phosphate groups as well as cholesterol and cholesterol esters in kidneys of rats exposed to IONPs may indicate that the particles stimulated formation of lipid droplets within the organ.

The intensity of the neutral environment impact on humans may be determined by specific features of space, including dead wood occurrence. Dead wood is claimed to be disliked by the public because it reduces the scenic beauty and recreational values of the forest. The attractiveness of a forest with dead wood may be determined by its variants. Much is known about the preference for landscape with dead wood, but there is little information available about how such a landscape affects a person’s mental relaxation, improves mood, increases positive feelings, levels of vitality, etc. Hence, the aim of our research was to investigate the psycho-logical relaxing effects of short 15-min exposures to natural and managed forests with dead wood. In the study, three areas within the Białowieża Primeval Forest were used to measure the impact of different types of forest with dead wood (A: forest reserve with dead wood subject to natural decomposition processes, B: managed forest with visible cut wood and stumps, C: man-aged forest with dead trees from bark beetle outbreak standing) on human psychological relaxation in a randomized experiment. The participants of the experiment were forty-one young adults aged 19–20. Each respondent experienced each type of forest at intervals visiting it. Four psychological questionnaires were used in the project (Profile of Mood States (POMS), Positive and Negative Affect Schedule (PANAS), Subjective Vitality Scale (SVS), and Restorative Outcome Scale (ROS)) before and after the short exposure to the forest were evaluated. The results show that a forest landscape with dead wood affects the human psyche, and the relaxing properties of such a landscape are better in a protected forest with natural, slow processes of tree dieback than those obtained in managed forests

This paper presents the results of research that aims to analyse and evaluate the attractiveness of the forest landscape in the vicinity of selected motor vehicle routes that differ in terms of technical parameters, such as motorways, regional roads and local (municipal) roads. The diversity of landscape units was used as the measure of landscape attractiveness. Landscape diversity analysis allows to establish the details related to specific parts of the road and the options for configuring the road environment. These options relate to the way the forest is managed and mainly involve aspects of infrastructure and cultivation. They include activities such as afforestation work and rebuilding the species composition of forest stands. According to the research, the lower the technical class of a road, the greater is the need to take action to adapt the structural-tree stand system to meet the safety requirements of road users.

Background and Objectives: Physical activity, recreation and walks successfully counteract negative symptoms of stress in people, especially in large cities, and have many positive psychological and physiological effects. There are many studies showing that contact with nature plays an important role in the regeneration of the human body. The city is not without green enclaves such as forests, parks or greenery along the streets. However, it is not entirely clear how the different physical characteristics of the urban space affect mood improvement, increase of positive feelings, vitality level, etc. Materials and Methods: In the study, two urban environments (apartment and green suburbs) were used, as well as two forests (coniferous and deciduous) to measure the impact of these environments on human physiological and psychological relaxation during a walk in a randomized experiment. The participants of the experiment were 75 young adult Poles studying in the largest Polish agglomeration, Warsaw. Before each experiment, the physiological and psychological state of the participant was measured indoors (pre-test). Four psychological questionnaires were used in the project (Profile of Mood States, Positive and Negative Affect Schedule, Restorative Outcome Scale, Subjective Vitality Scale), and physiological measurements (heart rate, blood pressure) before and after the short walking program were evaluated. Results: As a result of the analyses, it was shown that both staying in an urban environment with greenery and staying in a forest environment have a positive effect on the physiological and psychological relaxation of the subjects. A short walk in the suburbs was no less attractive than a walk in the forest in fall. The above indicates that various places with urban vegetation can be successfully used for recreation, just as in a forest where forest bathing is practiced. This indicates that different places with urban greenery can be successfully used for recreation, as can the forests where forest bathing is carried out.

The literature showing how age of humans or animals influences the IR absorption spectra recorded in different brain regions is very poor. A very limited number of studies used FTIR microspectroscopy for analysis of the aging process, however there is lack of data concerning the biomolecular changes occurring in the course of postnatal development of the central nervous system. Therefore, in this paper the topographic and semiquantitative biochemical changes occurring within the rat hippocampus during postnatal development were examined. To achieve the goal of the study, three groups of normal male rats differing in age were investigated. These were 6, 30, and 60 day old animals, and the chosen ages correspond to the neonatal period, childhood, and early adulthood in humans, respectively. Already, preliminary topographic analysis identified a number of significant changes in the accumulation of biomolecules within the hippocampal formation occurring during brain development. Such observation was confirmed by further semiquantitative analysis of intensities of selected absorption bands or ratios of their intensities. The detailed examinations were done for four hippocampal cellular layers (multiform, molecular, pyramidal, and granular layers), and the results showed that the accumulation of most biomolecules, including both saturated and unsaturated lipids as well as compounds containing phosphate and carbonyl groups, was significantly higher in adulthood comparing to the neonatal period. What is more, the increases in their levels were observed mostly between 6th and 30th days of animals' life. The unsaturation level of lipids did not change during postnatal development, although the differences in unsaturated and saturated lipids contents were noticed between examined animal groups. Significant differences in relative secondary structure of proteins were found between young adult rats and animals in neonatal period for which the relative level of proteins with β-type secondary structure was the highest.

Background In the brain, inflammation occurs following a variety of types of brain damage, including epileptic seizures. Proinflammatory cytokines, like IL-1β or TNFα, can increase neuronal excitability and initiate spontaneous seizures or epileptogenesis. Recent studies indicate that the effects can be attenuated or even abolished in animals subjected to inflammation-inducing treatments at earlier developmental stages, termed “preconditioning”. Immunocompetent microglial cells display particular sensitivity to subtle brain pathologies showing a morphological continuum from resting to reactive forms. Following inflammation, multiple ramified processes of resting microglia become gradually shorter, and the cells transform into macrophages. Parameters of the morphological variations were used here as indicators of the nervous tissue reactivity to seizures in adult rats experiencing inflammation at earlier stages of postnatal development. Methods Systemic inflammation was induced with lipopolysaccharide (LPS) in 6-day-old or 30-day-old rats. In two-month-old survivors of the inflammatory status, seizures were evoked with pilocarpine injection. The seizure intensity was scored during a six-hour continuous observation period following the injection. Brain sections were immunostained for Iba1 to visualize microglia. Thereafter, morphology of microglial cells located in the hippocampal formation was analyzed using parameters such as solidity, circularity, ramification index, and area. Results In naive rats, seizure-induced transformations of microglial cells were reflected by strong changes in the parameters of their morphology. However, in the adult rats pretreated with LPS on their 6th or 30th postnatal days, the seizure-induced changes were significantly reduced, and microglial morphology remained significantly closer to normal. Significant amelioration of the acute phase of seizures was observed only when inflammation was induced in 30-day-old, but not in 6-day-old, rats. Conclusions The results confirm previous reports that moderate inflammation protects the nervous tissue from subsequent damage by reducing influences of proinflammatory factors on reactive glial cells. The young-age inflammation may have age-dependent effects on susceptibility to seizures induced in adulthood. This article is part of a Special Issue entitled “Status Epilepticus”.

In this paper the elemental changes of rat hippocampal formation occurring during the postnatal development were examined. Three groups of animals were used in the study. These were naive Wistar rats at the age of 6-, 30- and 60-days and the chosen life periods corresponded to the neonatal period, childhood and early adulthood in humans, respectively. For the topographic and quantitative elemental analysis X-ray fluorescence microscopy was applied and the measurements were done at the FLUO beamline of ANKA. The detailed quantitative and statistical analysis was done for four areas of hippocampal formation, namely sectors 1 and 3 of the Ammon's horn (CA1 and CA3, respectively), dentate gyrus (DG) and its internal area (hilus of DG, H). The obtained results showed that among the all examined elements (P, S, K, Ca, Fe, Cu, Zn and Se), only the levels of Fe and Zn changed significantly during postnatal development of the hippocampal formation and both the elements were significantly higher in young adults comparing to the rats in neonatal period. The increased Fe areal density was found in all examined hippocampal areas whilst Zn was elevated in CA3, DG and H. In order to follow the dynamics of age-dependent elemental changes, the statistical significance of differences in their accumulation between subsequent moments of time was examined. The obtained results showed statistically relevant increase of Zn level only in the first observation period (between 6th and 30th day of life). Afterwards the areal density of the element did not change significantly. The increase of Fe areal density took place in both examined periods, however the observed changes were small and usually not statistically relevant.

Początki państwowej administracji i gospodarki leśnej sięgają VIII-IX wieku. Wraz z występującą w tym czasie wielką wędrówką ludów nastąpił znaczny wyrąb lasu na potrzeby ówczesnego osadnictwa. Kolejna nadmierna eksploatacja lasów w Europie miała miejsce w XIII-XV wieku i była wynikiem intensywnego rozwoju miast, rzemiosła, transportu, w tym drewnochłonnej floty oraz górnictwa, hutnictwa żelaza i szkła. Rosnące z roku na rok zużycie drewna doprowadziło do jego nadmiernego pozyskania i ubytku zasobności drzewostanów. Odzwierciedleniem tej sytuacji było zjawisko „głodu drewna”. Kolejny okres intensywnego użytkowania tego surowca nazwany został „epoką drewna” i trwał od XV do XIX wieku. Pierwszą usystematyzowaną teorią zajmującą się ekonomicznymi problemami gospodarki leśnej była kameralistyka (niem. Kammer − m.in. izba skarbowa), której początki rozwoju sięgają przełomu XVII i XVIII wieku. Jej nazwa pochodzi od byłych kamer rządowych. Kameralistów traktowano jako niemieckich przedstawicieli merkantylizmu i zaliczano do przedstawicieli liberalizmu gospodarczego. Wzrost zainteresowania leśnictwem ze strony kameralistów doprowadził do szybkiego rozwoju przemysłu leśnego. Za prekursora tych działań uważany jest W.G. Moser, autor dzieła „Zasady ekonomiki leśnictwa” (1757), według którego głównym celem leśnictwa jest zaspokojenie zapotrzebowania przemysłu na drewno1. Z obawy o brak dostatecznych dostaw drewna Moser formułuje „zasadę ciągłości użytkowania”, która w późniejszym okresie została określona jako „zasada ciągłości produkcji leśnej” i stała się kamieniem węgielnym rozwoju gospodarstwa leśnego. Na przełomie XVIII i XIX wieku gospodarkę leśną zaczyna kształtować liberalna ekonomia A. Smitha, wraz z jej wolnym rynkiem. Podstawowe jej zasady, w tym dotyczące zwłaszcza równowagi ekonomicznej gospodarstwa leśnego, znalazły swoje miejsce pod skrzydłami statyki leśnej. Jej wybitny przedstawiciel, G. Heyer (1865), określił ją natomiast jako rachunek oceny rentowności metod gospodarowania w leśnictwie2. To właśnie pod wpływem statyki żądano od leśnictwa dostarczenia „możliwie dużego dochodu pieniężnego” (zysku, renty), a nie największej masy drewna. Rozległość zainteresowania statyki leśnej doprowadziła do sformułowania dwóch podstawowych teorii ekonomiki leśnictwa określanych jako: ■ teoria renty gruntowej oraz ■ teoria renty leśnej. Pierwsza z wymienionych teorii główną uwagę koncentrowała na gruntach leśnych, które traktowała jako kapitał trwały gospodarstwa leśnego. Natomiast drzewostan był uznawany za kapitał (środek) obrotowy. Pojęcie gospodarności sprowadzało się do uzyskania maksymalnej rentowności mierzonej stosunkiem czystego (netto) dochodu (renty) do wartości kapitału trwałego gruntu leśnego. W obliczeniach tych statyka leśna korzystała w szerokim zakresie i nadal korzysta z rachunków procentu składanego. Wymieniona teoria renty leśnej nazywana również szkołą renty leśnej lub nauką o czystym dochodzie z lasu, dążyła do maksymalizacji czystego dochodu z lasu obejmującego przede wszystkim grunt i drzewostan. Wiek, w którym ten postulat był spełniony, określono wiekiem najwyższej renty leśnej lub wiekiem rębności. Teoretycznie wiek ten jest znacznie wyższy niż wiek rębności najwyższej renty gruntowej. Dla gorszych siedlisk różnice te wynosiły 40-60 i więcej lat, dla najlepszych zaś siedlisk około 30-40 lat. Wiek najwyższej rentowności był na ogół wyższy od wieku dojrzałości finansowej, a niższy od wieku najwyższej renty leśnej. W Europie Środkowej wiek rębności waha się od 60 do 250 lat (Möhring 2004)3. Długi okres produkcji leśnej na ogół dominuje w ekstensywnych metodach produkcji. Natomiast aktywne metody zagospodarowania lasu i jego pielęgnacji obejmują rocznie zaledwie od 10 do 20% ogółu powierzchni gospodarstwa leśnego. Oznacza to, że przeciętny cykl zabiegów hodowlano-ochronnych powtarzany jest co 5−10 lat. Przedstawiona krótka charakterystyka teorii statyki leśnej wskazuje na wysoki stopień jej operacjonizmu wynikający z empirycznych (fizycznych) metod pomiaru elementów rachunku renty gruntowej i renty leśnej. Pod tym względem rachunek rent wyraźnie różni się od pozostałych rachunków ekonomicznych gospodarki leśnej. Od XIX wieku, zwłaszcza od ukształtowania przez M. Faustmanna finansowego nurtu gospodarki leśnej, jej cele zostały zdominowane przez teorię czystego dochodu z gruntu, nazywanego również teorią renty gruntowej. Upowszechniła się ona po długim, przeszło 100-letnim sporze między nauką o czystym dochodzie z gruntu a nauką o czystym dochodzie z lasu. Obok teorii renty gruntowej i renty leśnej w XIX w. w leśnictwie zachodniej i środkowej części Europy ukształtowały się dwa nurty, które odbiegają pod względem celów i metod od teorii renty gruntowej i leśnej. Nurty te są bliżej określone w poniższych tytułach: ■ nurt ekologiczno-środowiskowy oraz ■ nurt ekonomiczno-finansowy. W Europie Środkowej i Zachodniej dominuje nurt ekologiczno-środowiskowy. Jego cechą są drzewostany wielogatunkowe o długim cyklu produkcji leśnej, słabych trzebieżach, niskim oprocentowaniu i małych odsetkach oraz o znacznym podporządkowaniu gospodarki leśnej czystemu dochodowi z lasu. Kolejną cechą nurtu ekologicznego jest duży przyrost miąższości, ale także znaczny udział drzewostanów starszych klas wieku w strukturach lasu oraz ich odporność na działanie biotycznych i abiotycznych zakłóceń, dywersyfikacja dochodów i ukierunkowanie celu gospodarki leśnej na maksymalizację czystego dochodu z lasu. Przeciwieństwem powyższego nurtu jest ekonomiczno-finansowa orientacja gospodarki leśnej ukierunkowana na krótkie wieki rębności, intensywne trzebieże, drzewostany jednogatunkowe, duże oprocentowanie i najwyższy czysty dochód z gruntu. Jednym słowem gospodarstwo o orientacji finansowo-ekonomicznej dąży do krótkich wieków rębności, zrębowego użytkowania oraz wysokich procentów. Tymczasem nurt ekologiczny preferuje długie wieki rębności i niskie oprocentowanie. Podsumowując należy zauważyć, że ukształtowane w przeszłości dwa nurty gospodarki leśnej dominują nadal w leśnictwie europejskim z różnym nasileniem poszczególnych metod. Ekonomiczne podejście do podejmowania decyzji inwestycyjnych w leśnych warunkach niepewności ryzyka zostało wsparte przez rozwijaną od roku 1950 teorię portfela, najpierw w Stanach Zjednoczonych Ameryki, później w innych krajach, w tym zwłaszcza w Skandynawii i Niemczech. Z jednej strony preferowane są więc drzewostany o dużej odporności na ryzyko, ale mniejszej produkcyjności, z drugiej zaś drzewostany o znacznej produkcyjności, ale małej odporności na ryzyko. Optymalna kombinacja drzewostanów reprezentujących te dwie cechy w jednym z przykładowych leśnictw w Niemczech miała następujący skład gatunkowy: 70% Św, 30% Bk (Knoke 2004)4. Natomiast w Szwecji optymalna struktura gatunkowa drzewostanów wynosi: 49% Św oraz 51% So (Svensson 2000)5. Do szczególnie ważnych i szeroko stosowanych obecnie rozwiązań należy dywersyfikacja, która polega na zmniejszeniu sumarycznego ryzyka przy jednakowym dochodzie lub zwiększeniu dochodu przy jednakowym ryzyku. Wynika stąd, że teoria portfela obejmuje kombinację dwóch dóbr, na przykład drzewosfery jodłowe i bukowe. Metodyczne aspekty omawianej teorii portfela są dziełem H. Markowitza (1952), który został za nie uhonorowany nagrodą Nobla w 1990 r.6 Natomiast pionierskiezastosowania teorii portfela w praktyce leśnictwa USA są wynikiem intensywnych prac W.L. Millsa i W.L. Hoovera (1982) nad rozwojem inwestycji leśnych7. Odnotowany w kolejnych dekadach dalszy rozwój teorii portfela dotyczy przede wszystkim leśnictwa Skandynawii oraz Niemiec. Zgodnie z teorią portfela jego istota polega na optymalnej kombinacji dwóch dóbr, na przykład dwóch gatunków w drzewostanie (Kruschwitz 2007)8. Powszechnym przykładem takiej kombinacji jest Las Tharandzki oraz sporządzone dla niego optymalne kombinacje struktury gatunkowej drzewostanów. Taki sam atrybut odnosi się również do innych kategorii dóbr. Dziewiąta edycja wspólnego przedsięwzięcia Instytutu Badawczego Leśnictwa i Dyrekcji Generalnej Lasów Państwowych, jakim jest Zimowa Szkoła Leśna, dotyczyła właśnie wyzwań stojących przed współczesnym leśnictwem, zarówno tych wynikających ze zmian globalnych środowiska, jak i ekonomiczno-społecznych aspektów leśnictwa. W IX Sesji uczestniczyło 259 osób. Poza autorami krajowymi, reprezentującymi Ministerstwo Środowiska, Dyrekcję Generalną Lasów Państwowych oraz jednostki organizacyjne Lasów Państwowych, uczelnie i instytuty naukowe, referaty wygłosili naukowcy i leśnicy praktycy z zagranicy. ----------------------------------------- 1 Moser W.G. 1757. Grundsätze der Forstökonomie. Bd. 1-2. Frankfurt a/M. 2 Heyer G. 1865. Anleitung zur Waldwertrechnung. Leipzig. 3 Möhring B. 2004. Betriebswirtschaftliche Analyse des Waldumbaus. Forst und Holz, 59, 11: 523-530. 4 Knoke T. 2004. Die Begründung von Mischbeständen: eine Möglichkeiten zur Minderung von Risiko? Kongressbericht des Deutschen Forstvereins zur Tagung 2003 in Mainz: 344-355. 5 Svensson L. 2000. Open-economy inflation targeting. Journal of International Economics, 50: 155–183. 6 Markowitz H. 1952. Portfolio Selection. The Journal of Finance, 7, 1: 77–91. 7 Mills W.L., Hoover W.L. 1982. Investment in forest land: aspects of risk and diversification. Land Economics, 58: 33-51. 8 Kruschwitz L. 2007. Finansowanie i inwestycje. CeDeWu, Warszawa.

Malformations of cortical development (MCD) are a common cause of intractable seizures in humans. Among these, focal cortical dysplasia (FCD) poses an outstanding challenge. There are several subtypes of FCD that show significant variation in pathology and clinical presentation. All types exhibit disturbed cortical cytoarchitecture and increased propensity for seizures. The etiology is likely heterogenous, with mutations, specifically in genes related to mammalian target of rapamycin (mTOR), identified in only a subset of cases. A more complex mechanism, in which underlying genetic background interacts with early, pre- or perinatal injury or stress, has been proposed. Here, we used a well-established animal model of developmental malformations similar to MCD, induced by prenatal gamma irradiation. Previously, a significant variation between times of treatment has been shown, resulting in distinct and lasting patterns of dysplasia and differentially altered seizure propensity. We set out to describe the molecular background of these patterns by performing microarray analyses of hippocampal samples obtained from adult rats previously irradiated at distinct time points during gestation: E13, E15, E17 or E19 as well as controls. The analysis was performed in three conditions: naïve, during latent phase after pilocarpine-induced status epilepticus and after 21 days of transauricular electric shocks. A set of 22 transcripts, some with known functions related to brain development, epilepsy and reaction to injury, was found to be altered between these groups across all treatments. We discuss the functional implication of these molecular differences, in an attempt to provide broader temporal and developmental context. © 2018 Wiley Periodicals, Inc. Develop Neurobiol 78: 718-735, 2018.

Astrocytes participate in neuronal development and excitability, and produce factors enhancing or suppressing inflammatory processes occurring due to neurodegenerative diseases, such as epilepsy. Seizures, in turn, trigger the release of inflammatory mediators, causing structural and functional changes in the brain. Therefore, it appears reasonable to determine whether generalized inflammation at developmental periods can affect astrocyte reactivity to epileptic seizures occurring in the adult brain. Lipopolysaccharide (LPS) was injected in 6- or 30-day-old rats (P6 or P30, respectively). At the age of 2 months, seizures were induced, and pilocarpine and morphological changes of astrocytes located within the hippocampal formation were assessed. Additionally, expression of glial fibrillary acidic protein (GFAP), glutamine synthetase (GS), aquaporin 4 (AQP4), and inwardly rectifying potassium channel Kir 4.1 (Kir4.1) was determined using Western blots. The animal group given LPS on P6 displayed maximal susceptibility to pilocarpine-induced seizures, significantly higher than the group that received LPS on P30. In the immunohistologically examined hippocampal formation, the GFAP-immunoreactive area was not affected by LPS alone. However, it was reduced following seizures in naive controls but not in LPS-pretreated rats. Increases in the ramification of astrocytic processes were detected only in adult rats given LPS on P30, not on P6. Seizures abolished the effects. Following seizures, the process ramification showed no significant change in the two LPS-treated rat groups, whereas it was significantly reduced in the dentate gyrus of LPS-untreated controls. Glial fibrillary acidic protein (GFAP) expression showed no changes induced with LPS alone and rose slightly after seizures. AQP4 content was lower in rats given LPS on P6 and was seizure-resistant in the two LPS-treated groups, contrary to a decrease in untreated controls. GS expression was not affected by LPS treatments and was reduced after seizures without an intergroup difference. Kir4.1 underwent highly significant increases in all groups experiencing seizures, but LPS alone had no effect. It can be concluded that the generalized inflammatory status led to some important changes in astrocytes reflected, in part at least by permanent modifications of their morphology and molecular profile. Moreover, the previously experienced inflammation prevented the cells from much stronger changes in response to seizures observed in adult untreated controls. The obtained results point to a link between the activation of astrocytes by transient systemic inflammation occurring during the developmental period and their subsequent reactivity to seizures, which may play an important role in the functional features of the brain, including its susceptibility to seizures.

This study aimed to estimate the prevalence of gastrointestinal nematodes and the intensity of infection in grazing dairy cattle from small and medium-sized farms in southern Poland. The level of antibodies against Ostertagia ostertagi in the bulk tank milk (BTM) from the animals was also assessed. Rectal fecal samples collected from 361 cows on 20 farms were examined using Willis-Schlaaf flotation and the McMaster method. BTM samples were tested for the presence of O. ostertagi antibodies using ELISA. Multiplex PCR was used to identify the third-stage larvae (L3) of gastrointestinal nematodes derived from the culture of pooled fecal samples from sampled farms. Gastrointestinal nematode eggs were found in the samples from 18 of the 20 herds with a prevalence range from 20.4 to 94.5%. The average number of eggs excreted in the feces of the herds was 200 eggs per gram (EPG). Antibodies to O. ostertagi were found in 20 of the examined herds (100%), of which 6 had optical density ratios (ODR) greater than 0.5. PCR results showed the presence of three nematode species: Ostertagia ostertagi, Cooperia oncophora and Oesophagostomum radiatum.

The main goal of this study was to evaluate in vivo effects of low dose of PEG-coated magnetic iron oxide nanoparticles (IONPs) on the rat liver. The IONPs was intravenously injected into rats at a dose equaled to 0.03 mg of Fe per 1 kg of an animal body weight. The elemental composition of liver tissue in rats subjected to IONPs action and controls were compared. Moreover, in order to determine the dynamics of nanoparticles (NPs) induced elemental changes, the tissues taken from animals 2 hours, 24 hours, and 7 days from IONPs injection were examined. The analysis of subtle elemental anomalies occurring as a result of IONPs action required application of highly sensitive analytical method. The total reflection X-ray fluorescence spectroscopy perfectly meets such requirements and therefore it was used in this study. The obtained results showed increasing trend of Fe level within liver occurring 2 hours from IONPs injection. One day after NPs administration, the liver Fe content presented the baseline level what suggests only the short-term accumulation of nanoparticles in the organ. The Ca, Cu, and Zn levels changed significantly as a result of NPs action. Moreover, the anomalies in their accumulation were still observed 7 days after IONPs injection. The level of Cu decreased while those of Ca and Zn increased in the liver of NPs-treated animals. The reduced liver Cu, followed by elevated serum level of this element, might be related in triggering the mechanisms responsible for Fe metabolism in the organism.

In this paper the influence of two different ketogenic diets (KDs) on the seizure-evoked elemental anomalies of hippocampal formation was examined. To achieve this purpose normal and pilocarpine treated rats previously fed with one of the two high fat and carbohydrate restricted diets were compared with animals on standard laboratory diet. The ketogenic ratios of the examined KDs were equal to 5:1 (KD1) and 9:1 (KD2). KD1 and standard diet fed animals presented similar patterns of seizure-evoked elemental changes in hippocampal formation. Also the analysis of behavioral data recorded after pilocarpine injection did not show any significant differences in intensity and duration of seizures between KD1 and standard diet fed animals. Higher ketogenic ratio KD2 introduced in the normal hippocampal formation prolonged changes in the accumulation of P, K, Zn and Ca. Despite this, both the intensity and duration of seizures were significantly reduced in rats fed with KD2 which suggests that its saving action on the nerve tissue may protect brain from seizure propagation. Also seizure-evoked elemental anomalies in KD2 animals were different than those observed for rats both on KD1 and standard diets. The comparison of seizure experiencing and normal rats on KD2, did not show any statistically significant differences in elemental composition of CA1 and H hippocampal areas whilst in CA3 area only Zn level changed as a result of seizures. DG was the area mostly affected by seizures in KD2 fed rats but areal densities of all examined elements increased in this hippocampal region.

In the present work, synchrotron radiation Fourier-transform infrared (SRFTIR) and Raman microspectroscopies were used to evaluate a possible role of creatine in the pathogenesis and progress of pilocarpine-evoked seizures and seizure-induced neurodegenerative changes in the rat hippocampal tissue. The main goal of this study was to identify creatine deposits within the examined brain area, to analyze their frequency in epileptic animals and naive controls and to examine correlations between the number of inclusions in the hippocampal formation of epileptic rats and the quantitative parameters describing animal behavior during 6-h observation period after pilocarpine injection. The presence of creatine in the brain tissue was confirmed based on the vibrational bands specific for this compound in the infrared and Raman spectra. These were the bands occurring at the wavenumbers around 2800, 1621, 1398, and 1304 cm−1 in IR spectra and around 1056, 908 and 834 cm−1 in the Raman spectra. Creatine was detected in eight of ten analyzed epileptic samples and in only one of six controls under the study. The number of deposits in epileptic animals varied from 1 to 100 and a relative majority of inclusions were detected in the area of the Dentate Gyrus and in the multiform hippocampal layer. Moreover, the number of creatine inclusions was positively correlated with the total time of seizure activity. Electronic supplementary material The online version of this article (doi:10.1007/s00216-011-5488-z) contains supplementary material, which is available to authorized users.

In the present work, synchrotron radiation Fourier transform infrared (SRFTIR) micro-spectroscopy and imaging were used for topographic and semi-quantitative biochemical analysis of rat brain tissue in cases of pilocarpine-induced epilepsy. The tissue samples were analyzed with a beam defined by small apertures and spatial resolution steps of 10 microm which allowed us to probe the selected cellular layers of hippocampal formation. Raster scanning of the samples has generated 2D chemical cartographies revealing the distribution of proteins, lipids and nucleic acids. Spectral analysis has shown changes in the saturation level of phospholipids and relative secondary structure of proteins. Special interest was put in the analysis of two areas of the hippocampal formation (sector 3 of the Ammon's horn, CA3 and dentate gyrus, DG) in which elemental abnormalities were observed during our previous studies. Statistically significant increase in the saturation level of phospholipids (increased ratio of the absorption intensities at around 2921 and 2958 cm(-1)) as well as conformational changes of proteins (beta-type structure discrepancies as shown by the increased ratio of the absorbance intensities at around 1631 and 1657 cm(-1) as well as the ratio of the absorbance at 1548 and 1657 cm(-1)) were detected in pyramidal cells of CA3 area as well as in the multiform and molecular layers of DG. The findings presented here suggest that abnormalities in the protein secondary structure and increases in the level of phospholipid saturation could be involved in mechanisms of neurodegenerative changes following the oxidative stress evoked in brain areas affected by pilocarpine-induced seizures.

A growing body of evidence demonstrates that dietary therapies, mainly the ketogenic diet, may be highly effective in the reduction of epileptic seizures. All of them share the common characteristic of restricting carbohydrate intake to shift the predominant caloric source of the diet to fat. Catabolism of fats results in the production of ketone bodies which become alternate energy substrates to glucose. Although many mechanisms by which ketone bodies yield its anticonvulsant effect are proposed, the relationships between the brain metabolism of the ketone bodies and their neuroprotective and antiepileptogenic action still remain to be discerned. In the study, X-ray fluorescence microscopy and FTIR microspectroscopy were used to follow ketogenic diet-induced changes in the elemental and biochemical compositions of rat hippocampal formation tissue. The use of synchrotron sources of X-rays and infrared allowed us to examine changes in the accumulation and distribution of selected elements (P, S, K, Ca, Fe, Cu, Zn, and Se) and biomolecules (proteins, lipids, ketone bodies, etc.) with the micrometer spatial resolution. The comparison of rats fed with the ketogenic diet and rats fed with the standard laboratory diet showed changes in the hippocampal accumulation of P, K, Ca, and Zn. The relations obtained for Ca (increased level in CA3, DG, and its internal area) and Zn (decreased areal density in CA3 and DG) were analogous to those that we previously observed for rats in the acute phase of pilocarpine-induced seizures. Biochemical analysis of tissues taken from ketogenic diet-fed rats demonstrated increased intensity of absorption band occurring at 1740 cm(-1), which was probably the result of elevated accumulation of ketone bodies. Moreover, higher absolute and relative (3012 cm(-1)/2924 cm(-1), 3012 cm(-1)/lipid massif, and 3012 cm(-1)/amide I) intensity of the 3012-cm(-1) band resulting from increased unsaturated fatty acids content was found after the treatment with the high-fat diet. This article is part of a Special Issue entitled "Status Epilepticus".

There is growing evidence that physical activity ameliorates the course of epilepsy in animal models as well as in clinical conditions. Since traumatic brain injury is one of the strongest determinants of epileptogenesis, the present study focuses on the question whether a moderate long-term physical training can decrease susceptibility to seizures evoked following brain damage. Wistar rats received a mechanical brain injury and were subjected to daily running sessions on a treadmill for 21 days. Thereafter, seizures were induced by pilocarpine injections in trained and non-trained, control groups. During the acute period of status epilepticus, the intensity of seizures was assessed within the six-hour observation period. The trained rats showed considerable amelioration of pilocarpine-induced motor symptoms when compared with their non-trained counterparts. Histological investigations of effects of the brain injury and of physical training detected significant quantitative changes in parvalbumin-, calretinin- and NPY-immunopositive neuronal populations. Some of the injury-induced changes, especially those shoved by parvalbumin-immunopositive neurons, were abolished by the subsequent physical training procedure and could, therefore, be considered as neuronal correlates of the observed functional amelioration of the injured brain.

In this study, ketogenic diet-induced biochemical changes occurring in normal and epileptic hippocampal formations were compared. Four groups of rats were analyzed, namely seizure experiencing animals and normal rats previously fed with ketogenic (KSE and K groups respectively) or standard laboratory diet (NSE and N groups respectively). Synchrotron radiation based Fourier-transform infrared microspectroscopy was used for the analysis of distributions of the main organic components (proteins, lipids, compounds containing phosphate group(s)) and their structural modifications as well as anomalies in creatine accumulation with micrometer spatial resolution. Infrared spectra recorded in the molecular layers of the dentate gyrus (DG) areas of normal rats on a ketogenic diet (K) presented increased intensity of the 1740 cm(-1) absorption band. This originates from the stretching vibrations of carbonyl groups and probably reflects increased accumulation of ketone bodies occurring in animals on a high fat diet compared to those fed with a standard laboratory diet (N). The comparison of K and N groups showed, moreover, elevated ratios of absorbance at 1634 and 1658 cm(-1) for DG internal layers and increased accumulation of creatine deposits in sector 3 of the Ammon's horn (CA3) hippocampal area of ketogenic diet fed rats. In multiform and internal layers of CA3, seizure experiencing animals on ketogenic diet (KSE) presented a lower ratio of absorbance at 1634 and 1658 cm(-1) compared to rats on standard laboratory diet (NSE). Moreover, in some of the examined cellular layers, the increased intensity of the 2924 cm(-1) lipid band as well as the massifs of 2800-3000 cm(-1) and 1360-1480 cm(-1), was found in KSE compared to NSE animals. The intensity of the 1740 cm(-1) band was diminished in DG molecular layers of KSE rats. The ketogenic diet did not modify the seizure induced anomalies in the unsaturation level of lipids or the number of creatine deposits.

Brain dysplasias produced by irradiation with gamma rays at various stages of prenatal development cause different post-natal susceptibility to seizures. To detect possible determinants of this difference, patterns of degenerative changes in the dysplastic brains following pilocarpine-induced epilepsy were analysed.Pregnant Wistar rats were exposed to a 1.0 Gy dose of gamma rays on gestation days 15 (E15) or 17 (E17). On post-natal day 60, their offspring received pilocarpine injections to evoke status epilepticus. Motor manifestations of seizure activity were observed continuously for 6 hours and rated. Six days following the status epilepticus, the rats were anesthetized and T(2)-weighted magnetic resonance (MR) images were obtained. Frontal sections of the brains were immunostained for immunoglobulins G (IgGs) to detect blood-brain barrier damage and IgG cell uptake and glial fibrillary acidic protein (GFAP) or S-100-beta protein to visualize astrocytes. Bandeiraea simplicifolia isolectin-B4 (BSI-B4) isolectin histochemistry was also performed to detect microglia/macrophages.Tissue damages within epileptic brains as observed by light microscopy generally reflected changes in magnetic resonance imaging (MRI) at similar locations. Brains of rats irradiated on E15 or E17 and showing epileptic symptoms at comparable intensity also displayed different distribution of the pathologic changes. Among other post-epileptic changes, in rats irradiated on E17 as well as controls, the laterodorsal and ventrolateral thalamic nuclei showed signs of severe degeneration. In rats irradiated on E15, the nuclei were free of such changes.The obtained data point to important differences in the pattern of propagation of epileptic activity in the dysplastic brains suffering from neuronal loss in functionally different structures.

Gliosis, axonal sprouting and remodelling of nerve connections in the injured brain have been regarded as epileptogenic processes dependent on the age when the injury was inflicted. The present study examines whether brains injured at different developmental stages may acquire different susceptibility to experimental status epilepticus induced in adulthood. In 6- and 30-day-old Wistar rats (P6s and P30s, respectively), a mechanical injury was performed in the left cerebral hemisphere. On postnatal day 60, all the animals and controls received single injections of kainic acid to evoke status epilepticus. During a 6-h period following the injection, the animals were observed continuously and motor symptoms of seizure activity were recorded and rated. P6s showed significantly lower intensity of kainic acid-induced epileptic symptoms and significantly longer survival than controls or P30s. In P30s, no significant change was detected. The data provide evidence that the developmental stage when the brain is injured determines epileptogenecity of the lesion. However, a considerable discrepancy between these data and those obtained previously following pilocarpine administration in the same experimental models of brain injury shows that each of the two models of epilepsy may display different aspects of the same age-dependent process triggered by the brain injury.

Summary: Purpose: To determine whether brains irradiated at different stages of prenatal development also have different postnatal susceptibility to seizures evoked by pilocarpine. Methods: Pregnant Wistar rats were exposed to a single 1.0-Gy dose of gamma rays on gestation days 13, 15, 17, or 19 (E13, E15, E17, and E19, respectively). On postnatal day 60, their offspring received i.p. pilocarpine injections to evoke status epilepticus. Behavior of the animals was observed continuously for 6 h after the injection, and motor manifestations of seizure activity were rated, and survival times recorded. After 7-day survival, the animals were killed, and their brains were weighed. Results: The average brain weight of animals exposed to irradiation at earlier prenatal stages (E13 or E15) was significantly lower than that after irradiation on E17 or E19. However, effects of the irradiation on the susceptibility to pilocarpine-induced seizures were quite opposite. The intensity of status epilepticus evoked in rats irradiated on E13 or E15 was significantly lower than that in nonirradiated controls or in those irradiated on E17 or E19. Moreover, after irradiation on E13 or E15, survival of the animals was significantly higher in relation not only to other irradiated groups but also to the controls. Conclusions: The results suggest than the extent of neuronal deficit, even if relatively greater, cannot always lead to higher susceptibility of the dysplastic brain to seizures. Functional consequences of the deficit, even if its magnitude is relatively smaller but involving specific brain areas, appear to be critical for the epileptogenesis.

Background Obesity is a worldwide epidemic with more than 600 million affected individuals. Human studies have demonstrated some alterations in brains of otherwise healthy obese individuals and elevated risk of neurodegenerative disease of old age; these studies have also pointed to slightly diminished memory and executive functions among healthy obese individuals. Similar findings were obtained in animal models of obesity induced by high fat diet. On the other hand, low carbohydrate high fat diets are currently promoted for losing weight (e.g., Atkin’s style diets). However, the long-term effects of such diets are not known. Additionally, high fat diets leading to (mild) ketonemia were shown to improve brain function in elderly humans and in some animal models. Aim To evaluate the hypothesis that long-term use of a high fat diet was associated with decreases in spatial memory, smaller hippocampi and hippocampi metabolite concentrations in Wistar rats. Methods Twenty five male Wistar rats were put on high fat diet (HFD; 60% calories from fat, 30% from carbohydrates) on their 55th day of life, while 25 control male rats (CONs) remained on chow. Adequate levels of essential nutrients were provided. Both groups underwent memory tests in 8-arm radial maze at 3rd, 6th, 9th, and 12th month. 1H magnetic resonance spectroscopy was employed to measure concentrations of tNAA (marker of neuronal integrity) at one month and one year, whereas MRI was used to evaluate hippocampal volumes. Results Obese rats (OBRs) consumed similar amount of calories as CONs, but less proteins. However, their protein intake was within recommended amounts. Throughout the experiment OBRs had statistically higher concentrations of blood ketone bodies than CONs, but still within normal values. At post-mortem assessment, OBRs had 38% larger fat deposits than CONs (p

Female Wistar rats were exposed to a single 1.0 Gy dose of gamma radiation on gestational days 13, 15, 17 or 19 (E13, E15, E17 and E19, respectively). Their 8- and 16-day old male offsprings were injected with 3 H -thymidine and sacrificed 4 h after the injection. Brain sections were immunostained for S100β protein and subjected to autoradiography. Thereafter, the dorsal part of the hippocampal formation was examined microscopically and numbers and locations of proliferating astrocytes were recorded. Following prenatal irradiation, the intensity of astrocyte proliferation was considerably reduced, especially in the region of dentate gyrus. The reduction showed regular trend of changes being much stronger in brains irradiated on E19 than in those irradiated on E13. The changes, therefore, were related to the stage of brain development at which the irradiation was performed. A possible role of neuronal regulatory influence on the postnatal development of glial cells was discussed.

Pregnant Wistar rats were exposed to a single 1.0 Gy dose of gamma rays on gestational days 13, 15, 17 or 19 (E13s, E15s, E17s and E19s, respectively). A mechanical injury was made in the cerebral hemisphere of their 6 day-old male offsprings. The injured rats were injected with [3H] thymidine on day 1 or 2 after injury and killed 4 h after the injection. Brain sections were processed for BSI-B4 isolectin histochemistry, subjected to autoradiography and examined microscopically to record numbers of proliferating and unproliferating macrophages located within the region of injury. The total number of macrophages as well as number of their divisions were minimal in E13s then showed a regular increase in E15s and E17s, and reached its maximal level in brains irradiated on E19. The trend of changes was opposite to that showed by changes in the intensity of astrocyte proliferation [Z. Setkowicz, K. Janeczko, Effects of prenatal gamma-irradiation on the astrocyte proliferation in response to injury in the brain of 6-day-old rat, Brain Res. 803 (1998) 122-128.]. The recruitment and proliferation of macrophages and the astrocyte proliferation were regarded as reactive processes occurring under control of different regulatory mechanisms acting within the region of injury.

Pregnant Wistar rats were exposed to a single 1.0 Gy dose of gamma rays on gestational days 13, 15, 17 or 19 (E13, E15, E17 and E19, respectively). A mechanical injury was made in the cerebral hemisphere of their 6 day-old male offsprings. The injured rats were injected with [ 3 H ] thymidine on day 1 or 2 after injury and killed 4 h after the injection. Brain sections were immunostained for glial fibrillary acidic protein (GFAP) or S-100β protein, subjected to autoradiography and examined microscopically to record proliferating astrocytes. The intensity of astrocyte proliferation in response to injury showed a gradual decrease from the level maximal in brains irradiated on E13 to minimal in those irradiated on E19. The changes were regarded as being related to the stage of prenatal development when irradiation of the brain was performed.

Our previous studies carried out on the pilocarpine model of seizures showed that highly resolved elemental analysis might be very helpful in the investigation of processes involved in the pathogenesis of epilepsy, such as excitotoxicity or mossy fiber sprouting. In this study, the changes in elemental composition that occurred in the hippocampal formation in the electrical kindling model of seizures were examined to determine the mechanisms responsible for the phenomenon of kindling and spontaneous seizure activity that may occur in this animal model. X-ray fluorescence microscopy was applied for topographic and quantitative analysis of selected elements in tissues taken from rats subjected to repetitive transauricular electroshocks (ES) and controls (N). The detailed comparisons were carried out for sectors 1 and 3 of the Ammon’s horn (CA1 and CA3, respectively), the dentate gyrus (DG) and hilus of DG. The obtained results showed only one statistically significant difference between ES and N groups, namely a higher level of Fe was noticed in CA3 region in the kindled animals. However, further analysis of correlations between the elemental levels and quantitative parameters describing electroshock-induced tonic and clonic seizures showed that the areal densities of some elements (Ca, Cu, Zn) strongly depended on the progress of kindling process. The areal density of Cu in CA1 decreased with the cumulative (totaled over 21 stimulation days) intensity and duration of electroshock-induced tonic seizures while Zn level in the hilus of DG was positively correlated with the duration and intensity of both tonic and clonic seizures.