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The tachykinins, including their major representative substance P (SP), are an essential component of its non-adrenergic, non-cholinergic transmission of the myenteric plexus of the enteric nervous system. Hence, the understanding of the role that SP plays in the conduction of a wide variety of motility and secretory reflexes lies in the evidence-based understanding of the sites of its expression. In the present study, the presence, distribution and staining intensity of the SP-positive nerve structures in the myenteric plexus of the proximal and distal colon of the adult rat have been investigated by means of immunohistochemistry at the light microscopic level. We observed an abundance of intensely stained beaded SP-immunopositive fibres, surrounding the SP-immunonegative myenteric neuronal cell bodies in a basket-like manner. Fine bundles of SP-immunostained varicose fibres were also registered in the adjacent smooth muscle layers. The statistical analysis revealed discernible prevalence of the staining intensity in the proximal colon, compared to the distal one. In summary, our findings provide immunohistochemical evidence for the occurrence of abundant SP-immunopositive nerve structures in the colonic region of the rat. Taken together with previous morphological and pharmacological research the results of this study suggest that the SP-mediated neurotransmission is of physiological and likely of pathological significance for the examined region.

This study aimed to compare the nasofrontal region in metopic and non-metopic cranial series and to assess whether the persistent metopic suture is related to a specific morphology of this part of the cranium. For the purpose of the study, a total of 159 dry crania (control series n = 90 and metopic one n = 69) of contemporary adult males were scanned with a laser scanner. Digital morphometry was accomplished by recording the three-dimensional coordinates of eleven landmarks, 3 bilateral and 5 in the mid-sagittal plane, characterizing the nasofrontal region. Between these landmarks, 43 linear measurements were calculated as Euclidian distances and 25 triangles were constructed. The angles, areas and heights of these triangles were also computed. The results show that the metopic crania have a distinctive nasofrontal morphology as the significant differences are not in the forehead height, but mainly in its configuration and in the nasal bones dimensions. The metopic crania have significantly flattened glabella, broad interorbital distance and wider, shorter and less prominent nasal bones compared to the non-metopic ones.

The persistent metopic suture is an anatomic variation related to a specific skull configuration. Data on the proportionality of the neurocranium and the occurrence of additional variations when the metopic suture persists are still insufficient. This study presents a comparison between a metopic and a non-metopic homogenous cranial series. The most distinctive quantitative and qualitative characteristics of the skull configuration when the metopic suture persists were identified using data mining techniques. A total of 175 contemporary adult male skulls were investigated. The skulls were divided in two groups: a control series (n = 100) and a metopic series (n = 75). All skulls were scanned with a hand-held laser scanner CreaformVIUscan and polygonal models were created. For 150 of the skulls, industrial µCT scanning was performed using Nikon XT H 225 and volumetric images were generated. A total of 92 attributes, both quantitative (dimensions) and qualitative (anatomical variations of the skull), were recorded from the 3D models. The major distinctive peculiarities observed in the neurocranial configuration when the metopic suture persists concerned the dimensions of the frontal bone which was considerably shorter, wider and more convex compared to the control. The metopic skulls manifested wormian bones in the sphenoidal fontanelle, around the temporal squama and along the lambdoid suture. These supernumerary bones were considerably more common in the metopic series in comparison with the control. The most reliable model resulting from the data mining summarizes a total of five classification rules entirely based on quantitative characteristics of the frontal bone.

Mastoid Triangle (MT) is a triangle constructed between the landmarks porion, mastoidale and asterion. The aim of the study is to evaluate the sex differences in the MT size in Bulgarian adults and to develop discriminant functions for sex estimation based on the MT sides and area. The study was carried out on 148 head CT scans. A sample of 53 males and 53 females was used for development of discriminant functions, and a test sample of 21 males and 21 females was applied for their validation. Using the software InVesalius©, 3D models of the skulls were segmented and exported in STL format. The 3D coordinates of the landmarks porion, asterion and mastoidale were collected using the software MeshLab©. The MT sides, area and angles were calculated. The sex differences were assessed by the independent t-test. Bilateral differences were evaluated using the paired t-test. Univariate and multivariate discriminant function analyses were applied. The results showed that the MT sides and area differed significantly between both sexes. Sex differences were also established for the angle at mastoidale. Bilateral differences were found in males for the distance porion-mastoidale, which was significantly greater on the right side. The MT dimensions showed sufficient discriminating power for sex estimation among Bulgarians (up to 89%), and the total MT area proved to be the best single sex discriminating trait. The test sample corroborated the usefulness of the MT in sex estimation demonstrating similar or higher overall accuracy rates.

Objectives The sagittal suture (SS) is assumed to be an initial site for the commencement of cranial suture closure as well as the most frequent spot of isolated craniosynostosis. The present study aimed to inspect the reorganization of the SS at the microlevel to assess the relation between its closure and aging and to establish whether it could be used as a reliable indicator in age-at-death prediction. Materials and methods The SS was investigated in 68 dry contemporary adult male skulls of known age-at-death. An additional series of 20 skulls was used for verification. The skulls were scanned using a micro-computed tomography system. The SS closure degree was assessed along the three bone layers on cross-sectional tomograms by using a scoring scale. Results In the entirely open SS, the bone edges consist of compact bone and are widely separated. With SS maturation, the bone edges come into contact, and the remodeling process leads to a decrease in the sutural area and bone homogenization across all three layers. SS closure is an irregular process roughly related to aging, beginning in the early 20s, reaching its peak at about 30 years of age and abating in the late 40s. Discussion Although related to aging, SS closure is not a simple function of it. Rather, the underlying factors inducing and managing this process are multifaceted and complex. Although the etiology of SS maturation remains unclear, it is reasonable to use SS closure cautiously and only as a supportive method for age prediction.

This study aimed to investigate the frequency of squamous suture (SqS) obliteration, to estimate the involvement of the major calvarial sutures and those surrounding the temporal squama, and to inspect the neuro- and basicranium for deformities. A series of 211 dry skulls of contemporary adult males were macroscopically observed. The skulls with closed SqS were scanned using an industrial µCT system. Digital morphometry of the skulls with obliterated SqS was performed by recording the 3D coordinates of anatomic landmarks and calculation of linear distances, angles and indices. Obliteration of SqS was observed in 3 (1.42%) skulls. One skull showed bilateral SqS obliteration. The other two cases were unilateral, one right-sided and one left-sided. SqS obliteration seems to be co-ordinated with the closure of the parietomastoid suture, partially related to the closure of the occipitomastoid, sphenoparietal and sphenofrontal sutures, and independent from the closure of the sphenosquamosal suture and the major calvarial sutures. No severe disproportions in the skull configuration were observed in the three investigated cases. The major differences in the complimentary hemicrania concern the parietal and occipital parts of the skull vault. Dorsum sellae erosion, an indicator for raised intracranial pressure, was observed in all three cases.

Depression, concomitant with epilepsy, may cause serious impairment in one’s quality of life, not limited to seizures. Investigations of biomarkers and new therapeutic approaches targeting comorbid depression in epilepsy have been in progress for the past decade. In the attempt to find the mechanisms underlying the development of depression in epilepsy, an important part of the research is devoted to elucidating what the precise impact of inflammatory mediators is. Here, we overview the role of critical inflammatory pathways involved in the development of depression comorbid to epilepsy. The scarcity of evidence in literaure that has contributed to the elaboration of the hypothesis for a common and specific inflammatory signaling mechanism responsible for the generation of depressive state in epilepsy is discussed. In addition, the possibility of the implementation of suitable antiinflammatory therapies suggested to ameliorate depression accompanying epilepsy is analyzed.

The aim of the present study is to apply support vector machines (SVM) and artificial neural network (ANN) as sex classifiers and to generate useful classification models for sex estimation based on cranial measurements. Besides, the performance of the generated sub-symbolic machine learning models is compared with models developed through logistic regression (LR). The study was carried out on computed tomography images of 393 Bulgarian adults (169 males and 224 females). The three-dimensional coordinates of 47 landmarks were acquired and used for calculation of the cranial measurements. A total of 64 measurements (linear distances, angles, triangle areas and heights) and 22 indices were calculated. Two datasets were assembled including the linear measurements only and all measurements and index, respectively. An additional third dataset comprising all possible interlandmark distances between the landmarks was constructed. Two machine learning algorithms—SVM and ANN and a traditional statistical analysis LR—were applied to generate models for sex estimation. In addition, two advanced attribute selection techniques (Weka BestFirst and Weka GeneticSearch) were used. The classification accuracy of the models was evaluated by means of 10 × 10-fold cross-validation procedure. All three methods achieved accuracy results higher than 95%. The best accuracy (96.1 ± 0.5%) was obtained by SVM and it was statistically significantly higher than the best results achieved by ANN and LR. SVM and ANN reached higher accuracy by training on the full datasets than the selection datasets, except for the sample described by the interlandmark distances, where the reduction of attributes by the GeneticSearch algorithm improved the accuracy.

The aim of the present study is to develop effective and understandable classification models for sex estimation and to identify the most dimorphic linear measurements in adult crania by means of data mining techniques. Furthermore, machine learning models and models developed through logistic regression analysis are compared in terms of performance. Computed tomography scans of 393 adult individuals were used in the study. A landmark-based approach was applied to collect the metric data. The three-dimensional coordinates of 47 landmarks were acquired and used for calculation of linear measurements. Two datasets of cranial measurements were assembled, including 37 standard measurements and 1081 interlandmark distances, respectively. Three data mining algorithms were applied: the rule induction algorithms JRIP and Ridor, and the decision tree algorithm J48. Two advanced attribute selection methods (Weka BestFirst and Weka GeneticSearch) were also used. The best accuracy result (91.9 %) was achieved by a set of rules learnt by the JRIP algorithm from the dataset constructed by application of the GeneticSearch selection algorithm to the dataset of standard cranial measurements. The set consisted of five rules including seven cranial measurements. Its accuracy was even better than the classification rates achieved by the logistic regression models. Concerning the second dataset of nonstandard measurements, the best accuracy (88.3 %) was obtained by using classification models learnt by two algorithms – JRIP with a dataset preprocessed by the BestFirst selection algorithm and Ridor with preprocessing by the GeneticSearch selection algorithm. Our experiments show that for the two datasets mentioned above the rule-based models contain smaller sets of rules with shorter lists of measurements and achieve better classification accuracy results in comparison with decision tree-based models.

Foramen magnum (FM) has a well-protected position, which makes it of particular interest in forensic research. The aim of the study is to assess the sex differences in size and shape of FM, develop discriminant functions and logistic regression models based on the FM measurements, compare the accuracy results of the measurements obtained through different measuring approaches, and establish the most reliable variables for sex estimation in Bulgarian adults. Head CT scans of 140 Bulgarian adults were used in the study. The segmentation of the skulls was performed in the software InVesalius. The length, breadth, circumference, and area were measured based on the 3D coordinates of definite landmarks and semi-landmarks. The circumference and area were calculated regarding the foramen as a 2D and 3D structure. Two additional variables (λ2 and λ3) corresponding to the least square errors along the length and breadth directions at the fitting of the 3D coordinates to a plane were examined for their sex discriminating ability. The FM shape was classified based on the values of the FM index. The significance of the sex differences was assessed. Discriminate function analysis and binary logistic regression were conducted. Significant sex differences were established in the FM size and shape. The eigenvalue λ3 is the best discriminating parameter applying discriminant function analysis. The acceptance of FM as a 2D or 3D structure does not provide substantial information for its sex discrimination. The measurements of FM do not offer sufficiently high predicting rates for sex estimation in the Bulgarian population.

Background The morphology and dimensions of the frontal sinus (FS) are significant in the forensic, surgical and population context. Purpose The study aimed to compare linear FS measurements taken both on radiographs and virtual endocasts and to assess the impact of the skull angulation on the FS dimensions. Material and Methods Thirteen intact dry skulls of contemporary adult males were radiographed using industrial digital radiography while they were inclined in the Frankfurt plane, through the Caldwell's view up to the Water's view by angular steps of 5°. The width and height of both frontal lobes were measured in each projection. To verify the measurements on the radiographs, ten of the skulls were µCT-scanned and virtual endocasts of the FS were generated. Results The concordance between the measurements on the virtual endocasts and the radiographs in the Caldwell's view showed almost perfect concurrence for the width (0.998) and height on the left side (0.990), and substantial one for the height on the right side (0.961). Since the width is more sensitive compared to the height, any inclination from the initial position at the Caldwell's view caused a significant distortion of the FS measurements. Conclusion The industrial µCT-systems support both 2D and 3D imaging and could generate images with a high resolution. Therefore, if the industrial digital radiography is selected as an eligible imaging modality for FS investigation and documentation in conformity with the research goals, the appropriate skull positioning ensures reliable readings of the linear FS dimensions.

The carotid body (CB) is a major peripheral arterial chemoreceptor that initiates respiratory and cardiovascular adjustments to maintain homeostasis. Recent evidence suggests that circulating or locally produced hormones like angiotensin II acting via AT1 receptors modulate its activity in a paracrine-autocrine manner. The aim of this study was to examine the immunohistochemical localization of AT1 receptor in the CB of adult rats and to compare its expression in vehicle-treated animals, and after the long-term application of its selective blocker losartan. Immunohistochemistry revealed that a subset of CB glomeruli and the vast majority of neurons in the adjacent superior cervical ganglion (SCG) were strongly AT1 receptor-immunoreactive. In the CB immunostaining was observed in the chemosensory glomus cells typically aggregated in cell clusters while the nerve fibers in-between and large capillaries around them were immunonegative. Exogenous administration of losartan for a prolonged time significantly reduces the intensity of AT1 receptor immunostaining in the CB glomus cells and SCG neurons. Our results show that AT1 receptors are largely expressed in the rat CB under physiological conditions, and their expression is down-regulated by losartan treatment.

Aims Experimental and clinical studies have demonstrated that components of renin-angiotensin system are elevated in the hippocampus in epileptogenic conditions. In the present work, we explored the changes in the expression of angiotensin II receptor, type 1 (AT1 receptor) in limbic structures, as well as the effect of the AT1 receptor antagonist losartan in a model of comorbid hypertension and epilepsy. Main methods The expression of AT1 receptors was compared between spontaneously hypertensive rats (SHRs) and Wistar rats by using immunohistochemistry in the kainate (KA) model of temporal lobe epilepsy (TLE). The effect of losartan was studied on AT1 receptor expression in epileptic rats that were treated for a period of 4 weeks after status epilepticus. Key findings The naive and epileptic SHRs were characterized by stronger protein expression of AT1 receptor than normotensive Wistar rats in the CA1, CA3a, CA3b, CA3c field and the hilus of the dentate gyrus of the dorsal hippocampus but fewer cells were immunostained in the piriform cortex. Increased AT1 immunostaining was observed in the basolateral amygdala of epileptic SHRs but not of epileptic Wistar rats. Losartan exerted stronger and structure-dependent suppression of AT1 receptor expression in SHRs compared to Wistar rats. Significance Our results confirm the important role of AT1 receptor in epilepsy and suggest that the AT1receptor antagonists could be used as a therapeutic strategy for treatment of comorbid hypertension and epilepsy.

Recent studies about the novel antidepressant agomelatine, which is a mixed MT1 and MT2 melatonin receptor agonist and 5HT2C serotonin receptor antagonist possessing an anticonvulsant and neuroprotective action, suggest that it may have potential to contribute against epileptogenesis and epilepsy-induced memory impairment. In order to ascertain whether protection of some brain structures could suppress epileptogenesis, in the present study, we evaluated the effect of chronic post-status treatment with agomelatine on epileptogenesis, behavioral and neuronal damage induced by kainate acid (KA) status epilepticus (SE). Agomelatine/vehicle treatment (40 mg/kg, i.p.) started one hour after SE and continued up to 10 weeks in Wistar rats. Latency for onset of spontaneous motor seizures (SMS) and their frequency was detected by a 24-h video-recording. Locomotor activity, anxiety and hippocampus-dependent spatial memory in open field (OF), elevated plus maze (EPM), light-dark test (LDT) and radial arm maze (RAM) test, respectively, were evaluated during the last two weeks after SE. Agomelatine significantly decreased the latency for onset of SMS and increased the seizure frequency during the 2nd and the 3rd week of treatment. The MT1 and MT2 receptor agonist and serotonin 5HT2C receptor antagonist exacerbated the KA-induced hyperlocomotion and impulsive behavior and it was unable to prevent spatial memory impairment of epileptic rats. However, agomelatine induced a neuroprotection in the dorsal hippocampus, specifically in the CA1, septal CA2 and partially in the CA3c region, the hilus of the dentate gyrus, piriform cortex and septo-temporal and temporal basolateral amygdala. Our findings suggest that the beneficial impact against SE-induced neuronal loss exerted by agomelatine is not crucial for the suppression of epileptogenesis and its deleterious consequences in KA model of temporal lobe epilepsy.

The carotid body (CB) is a multipurpose metabolic sensor that acts to initiate cardiorespiratory reflex adjustments to maintain homeostasis of blood-borne chemicals. Emerging evidence suggests that nitric oxide increases the CB chemosensory activity and this enhanced peripheral chemoreflex sensitivity contributes to sympathoexcitation and consequent pathology. The aim of this study was to examine by means of NADPH-diaphorase histochemistry and nitric oxide synthase (NOS) immunohistochemistry the presence and distribution of nitrergic structures in the CB of spontaneously hypertensive rats (SHRs) and to compare their expression patterns to that of age-matched normotensive Wistar rats (NWRs). Histochemistry revealed that the chemosensory glomus cells were NADPH-d-negative but were encircled by fine positive varicosities, which were also dispersed in the stroma around the glomeruli. The NADPH-d-reactive fibers showed the same distributional pattern in the CB of SHRs, however their staining activity was weaker when compared with NWRs. Thin periglomerular, intraglomerular and perivascular varicose fibers, but not glomus or sustentacular cells in the hypertensive CB, constitutively expressed two isoforms of NOS, nNOS and eNOS. In addition, clusters of glomus cells and blood vessels in the CB of SHRs exhibited moderate immunoreactivity for the third known NOS isoenzyme, iNOS. The present study demonstrates that in the hypertensive CB nNOS and eNOS protein expression shows statistically significant down-regulation whereas iNOS expression is up-regulated in the glomic tissue compared to normotensive controls. Our results suggest that impaired NO synthesis could contribute to elevated blood pressure in rats via an increase in chemoexcitation and sympathetic nerve activity in the CB.

This study aimed to investigate the frequency of squamous suture (SqS) obliteration, to estimate the involvement of the major calvarial sutures and those surrounding the temporal squama, and to inspect the neuro- and basicranium for deformities. A series of 211 dry skulls of contemporary adult males were macroscopically observed. The skulls with closed SqS were scanned using an industrial µCT system. Digital morphometry of the skulls with obliterated SqS was performed by recording the 3D coordinates of anatomic landmarks and calculation of linear distances, angles and indices. Obliteration of SqS was observed in 3 (1.42%) skulls. One skull showed bilateral SqS obliteration. The other two cases were unilateral, one right-sided and one left-sided. SqS obliteration seems to be co-ordinated with the closure of the parietomastoid suture, partially related to the closure of the occipitomastoid, sphenoparietal and sphenofrontal sutures, and independent from the closure of the sphenosquamosal suture and the major calvarial sutures. No severe disproportions in the skull configuration were observed in the three investigated cases. The major differences in the complimentary hemicrania concern the parietal and occipital parts of the skull vault. Dorsum sellae erosion, an indicator for raised intracranial pressure, was observed in all three cases.

The aim of the study was to determine the impact of the resolution and texture of three-dimensional (3D) models created through laser scanning on the measurement error (ME) of craniometric landmarks. Ten skulls were scanned at five different resolutions, and the generated 3D models were exported with and without texture. The 3D coordinates of 28 landmarks were derived. Each landmark was picked five times by one observer. The ME of a definite landmark was calculated as an average of distances between the repeated placements of the landmark by the observer and the landmark centroid. One-way analysis of variance was applied for detection of significant differences in the MEs between and within landmark types recorded at different resolutions. The MEs of landmark types in textured and nontextured models were compared by a paired test. Twelve linear measurements were calculated as interlandmark distances, and their values obtained on the models of different resolution were compared. The Frankfurt horizontal plane was constructed for each model and its deviation was calculated at different resolutions. Scan resolution impacted MEs of Type 1 and Type 2 landmarks but not the precision level of Type 3 landmarks. Texture most influenced the precise identification of Type 1 landmarks. The interlandmark distances between Type 2 landmarks were most consistent in their values, those between Type 1 landmarks showed deviations in low-resolution models, and distances between Type 3 landmarks demonstrated various patterns of transition of the values throughout the resolutions. Altogether, the use of textured high-resolution models would be preferable in morphometric studies. Anat Rec, 2019. © 2019 American Association for Anatomy Anat Rec, 303:1950-1965, 2020. © 2019 American Association for Anatomy.

Recently, we have reported that while agomelatine (Ago) is unable to prevent development of epilepsy it exerts a strong neuroprotective and anti-inflammatory response in the KA post-status epilepticus (SE) rat model. In the present study, we aimed to explore whether the brain-derived neurotrophic factor (BDNF) in the hippocampus is involved in the neuroprotective effect of Ago against the KA-induced SE and epileptiform activity four months later in rats. Lacosamide (LCM) was used as a positive control. The EEG-recorded seizure activity was also evaluated in two treatment protocols. In Experiment#1, Ago given repeatedly at a dose of 40 mg/kg during the course of SE was unable neither to modify EEG-recorded epileptiform activity nor the video- and EEG-recorded spontaneous seizures four months later compared to LCM (50 mg/kg). However, both Ago and LCM inhibited the expression of BDNF in the mossy fibers and also prevented neuronal loss in the dorsal hippocampal and the piriform cortex after SE. In Experiment#2, acute injection of Ago and LCM on epileptic rats, characterized by high seizure rates, did not prevent EEG-recorded paroxysmal events while only LCM decreased either absolute or relative powers of gamma (28–60 Hz) and high (HI) (60–120 Hz) frequency bands to baseline in the frontal and parietal cortex, respectively. Our results suggest that the protection against neuronal loss in specific limbic regions and overexpressed BDNF in the mossy fibers resulting from the repeated treatment with Ago and LCM, respectively, during SE is not a prerequisite for alleviation of epileptogenesis and development of epilepsy. In addition, a reduction of gamma and HI bands in the frontal and parietal cortex is not associated with EEG-recorded paroxysmal events after acute injection of LCM.

The carotid body (CB) is a small neural crest-derived chemosensory organ that detects the chemical composition of the arterial blood and responds to its changes by regulating breathing. The effects of acute nitrite treatment on the CB morphology in rats were examined by morphometry. We found that 1h after administrating a single dose of sodium nitrite, the CB underwent structural changes characterized by a prominent increase in its size with a marked, several-fold dilation of the blood vessels. The obvious CB enlargement mostly due to apparent vasodilation and glomus cell hypertrophy was at its highest one day later and persisted until the fifth day. 20 days after the treatment, the CB regained its size to the normoxic control state. Morphometric analysis revealed that the CB size increase in treated animals is statistically significant when compared to that of untreated controls. It can be inferred that the nitrite-exposed CB displays remarkable structural plasticity and enlarges its size mostly through vascular expansion.

Inflammatory signal molecules are suggested to be involved in the mechanism underlying comorbid depression in epilepsy. In the present study, we tested the hypothesis that the novel antidepressant agomelatine, a potent melatonin MT1 and MT2 receptor agonist and serotonin 5HT2C receptor antagonist, can prevent depressive symptoms developed during the chronic epileptic phase by suppressing an inflammatory response. Chronic treatment with agomelatine (40 mg/kg, i.p.) was initiated an hour after the kainate acid (KA)-induced status epilepticus (SE) and maintained for a period of 10 weeks in Wistar rats. Registration of spontaneous motor seizures was performed through a video (24 h/day) and EEG monitoring. Antidepressant activity of agomelatine was explored in the splash test, sucrose preference test (SPT) and forced swimming test (FST) while anxiolytic effect was observed through the novelty suppression-feeding test (NSFT) during chronic phase in epileptic rats. The frequency of motor seizures detected by video and EEG recording did not differ between vehicle and Ago group. Rats with registered spontaneous motor seizures showed symptoms typical for depressive behavior that included decreased grooming, anhedonia during the dark period and hopeless-like behavior. Epileptic rats exhibited also anxiety with novelty-induced hypophagia. This behavioral deficit correlated with increased signal markers of inflammation (plasma levels of interleukin (IL)-1β and activated glia in brain), while plasma corticosterone levels were not changed. Agomelatine treatment during epileptogenesis exerted a clear antidepressant effect by suppressing all behavioral hallmarks, reducing plasma IL-1β levels and preventing microgliosis and astrogliosis in specific limbic regions. The present results suggest that agomelatine treatment starting after SE can provide an effective therapy of comorbid depression in chronic epileptic condition through suppression of inflammatory signaling.

Mast cells (MCs) are important object in experimental acupuncture due to their putative involvement in local reactions to needling. In the rat, they are shown to contain in their granules, among other tissue mediators, serotonin, also called 5-hydroxytryptamine (5-HT). The aim of this study is to examine the normal distribution of 5-HT-containing MCs in soft tissues of Zusanli (ST36) acupuncture point (acupoint) and their morphological changes caused by experimental acupuncture. We observed 5-HT-immunopositive MCs in the tissues and in the vicinity of the needle tract formed after acupuncture. As a result of acupuncture needling, the tissue integrity is disrupted and certain folds are formed in the direction of the needle tract. Connective tissue in the vicinity of the needle tract gets compressed and displaced, together with the 5-HT-immunoreactive MCs seen there. Some of those 5-HT-immunopositive MCs showed signs of degranulation with numerous discharged granules, some of them found at a considerable distance form the cell. Furthermore, 5-HT-immunopositive MCs are unevenly distributed in soft tissues of ST36 acupoint. Larger numbers of 5-HT-containing MCs were visualized in subcutis and dermis, compared to the observed in striated muscles. Placing the acupuncture needle into the rat skin caused a formation of an apparent needle tract, tissue displacement and degranulation of 5-HT-immunopositive MCs. The demonstrated serotonin release by means of MC degranulation might be involved in the local tissue response to acupuncture.

The therapeutic efficacy of regular physical exercises in an animal model of epilepsy and depression comorbidity has been confirmed previously. In the present study, we examined the effects of endurance training on susceptibility to kainate (KA)-induced status epilepticus (SE), behavioral changes and neuronal damage in spontaneously hypertensive rats (SHRs). Male SHRs were randomly divided into two groups. One group was exercised on a treadmill with submaximal loading for four weeks and the other group was sedentary. Immediately after the training period, SE was evoked in half of the sedentary and trained rats by KA, while the other half of the two groups received saline. Basal systolic (SP), diastolic (DP) and mean arterial pressure (MAP) of all rats were measured at the beginning and at the end of the training period. Anxiety, memory and depression-like behaviour were evaluated a month after SE. The release of 5-HT in the hippocampus was measured using a liquid scintillation method and neuronal damage was analyzed by hematoxylin and eosin staining. SP and MAP of exercised SHRs decreased in comparison with the initial values. The increased resistance of SHRs to KA-induced SE was accompanied by an elongated latent seizure-free period, improved object recognition memory and antidepressant effect after the training program. While the anticonvulsant and positive behavioral effects of endurance training were accompanied by an increase of 5-HT release in the hippocampus, it did not exert neuroprotective activity. Our results indicate that prior exercise is an effective means to attenuate KA-induced seizures and comorbid behavioral changes in a model of hypertension and epilepsy suggesting a potential influence of hippocampal 5-HT on a comorbid depression. However, this beneficial impact does not prevent the development of epilepsy and concomitant brain damage.

Recently, we have shown that the blockade of AT1 receptor might be useful as an adjuvant treatment strategy for the prevention of oxidative stress and neurotoxicity caused by status epilepticus (SE) in rats. The purpose of the present study was to further assess the efficacy of long-term treatment with losartan (10mg/kg), the selective AT1 receptor antagonist, during kainate (KA)-induced epileptogenesis in Wistar rats. Losartan treatment started after onset of SE and continued for 4weeks. The rats were video- and EEG-recorded for 3months. Locomotor activity, anxiety and depressive-like behavior were evaluated 9weeks after SE, when all rats had developed chronic epileptic state. Neuronal damage in hippocampus was analyzed by hematoxylin while serotonin (5-HT) levels in hippocampus by HPLC. AT1 receptor antagonism increased the latent seizure-free period and decreased the frequency of spontaneous motor seizures. Losartan positively affected epilepsy-provoked behavioral changes, including impulsivity, low anxiety level and depression in a phase-dependent manner and restored the changes in diurnal fluctuation of motor activity. Losartan exerted neuroprotection selectively in the CA1 area of the hippocampus in the KA-treated rats and lowered the 5-HT levels both in normal and abnormal conditions. Our findings suggest that the AT1 receptor antagonist exerts disease-modifying effects during KA-induced epileptogenesis and neuronal damage in CA1 hippocampal area, attenuated some of the behavioral changes and restored diurnal variability in locomotor activity.

Background: The aim of the study is to measure the facial soft tissue thicknesses (STTs) in Bulgarians, to evaluate the relation of the STTs to the nutritional status, sex and bilateral asymmetry, and to examine the correlations between the separate STTs as well as between the STTs and body weight, height, and body mass index (BMI). In the present study, the facial STTs were measured on computed tomography scans of the head of Bulgarian adults. Materials and methods: The STTs were measured at 7 midline and 9 bilateral landmarks. The measurements were performed in the free software InVesalius in the axial and sagittal planes. The mean, standard deviation, minimum and maximum values, median and coefficient of variation were reported for the STT at each landmark according to the sex and BMI category. The BMI, sex and bilateral differences were assessed for statistical significance. Pearson correlation analysis was applied to assess the strength and direction of the relationships between the STTs and body height, weight and BMI, as well as between separate STTs. Results and Conclusions: The facial soft tissues in Bulgarian adults changed in accordance with the nutritional status of the individual and in both sexes all STTs augmented with the increasing BMI. For both normal and overweight BMI categories, males had more soft tissue at the majority of facial points than females, as the only exceptions were observed in the cheek zone, where STTs were thicker in females. Significant bilateral differences were observed in either sex and BMI category. Stronger correlations were established for the STTs in the jaw region and between the cheek and jaw soft tissues. Besides, the correlations between the homologous bilateral landmarks were among the strongest ones.

Experimental and clinical studies have demonstrated that components of renin-angiotensin system are elevated in the hippocampus in epileptogenic conditions. In the present work, we explored the changes in the expression of angiotensin II receptor, type 1 (ATThe expression of ATThe naive and epileptic SHRs were characterized by stronger protein expression of ATOur results confirm the important role of AT

OBJECTIVES This study aimed to establish the frequency of the frontal sinus (FS) aplasia, to compare metopic and nonmetopic series and thus to assess the relationship between the preservation of metopic suture and FS development. MATERIALS AND METHODS FSs were investigated in 230 dry skulls of adult males distributed into control (137) and metopic (93) series. They were visualized through industrial digital radiography. RESULTS In the control series, the FS aplasia was observed in 12.41% of the skulls, and it was mostly unilateral (8.76%) than bilateral (3.65%). The left-sided aplasia (5.11%) slightly prevailed over the right-sided one (3.65%). In the metopic series, the aplasia was observed with a frequency of 19.35%, and the bilateral aplasia (7.53%) was rarer that the unilateral one (11.83%), while the right-sided aplasia was clearly predominant (9.68%) compared to the left-sided one (2.15%). DISCUSSION The significant differences between both series showed a tendency for the persistence of metopic suture to be frequently related with FS underdevelopment in the vertical plate of the frontal bone, but in cases of pneumatization, it was preferentially on the left side. Taking into account that the cranial hypertension leads to suture diastasis and hinders development of the FS, it could be suggested that persistence of the metopic suture along with underdevelopment of the FS in nonsyndromic adults could be an expression of an elevated intracranial pressure during early development as an after-effect of certain condition.

Tripeptidyl peptidase I (TPPI) is a lysosomal enzyme widely distributed in mammals and humans. Its genetically determined deficiency causes the classical late-infantile form of neuronal ceroid lipofuscinosis, a fatal hereditary neurodegenerative disease associated with severe symptoms and early death, usually in the second decade of life. Many studies also show that TPPI is differentially regulated under various pathological conditions such as malignancy, neurodegeneration, ischemia, and inflammation, pointing at possible enzyme involvement in the pathogeneses of these entities. This chapter focuses on the TPPI participation in neurodegenerative and neoplastic diseases.

Obliteration of the cranial sutures is an age-dependent process. Its premature occurrence (craniosynostosis) causes different craniofacial deformations, dependent on the affected suture(s). The understanding of the suture morphology and the remodeling processes during the obliteration is essential for early diagnosis and treatment of the premature closure. This study aimed to investigate the morphology of open and obliterated sutures and to perform comparison analysis on the 3D images obtained by both industrial and medical computed tomography (CT) systems with various resolutions. A segment of the sagittal suture of dry skulls of known age and sex was scanned using Nikon XTH 225, an industrial CT system, developed by Nikon Metrology. The same section of the sagittal suture was observed on patients undergoing CT scanning with a multislice system Toshiba Aquilion 64 with 0.5 mm slice thickness. For 3D visualization, VGStudioMax 2.2 were used. The suture morphology was observed in coronal section on sequential 2D slices. Micro-CT (μCT) scanning of dry skulls enabled calculation of the morphometric parameters and visualization of the microarchitecture of the suture and its reorganization during the obliteration, unlike the CT imaging of patients, where the sutures were scarcely discernable. In the entirely open sections of the suture the bone edges were separated by a gap of various widths. As the obliteration proceeded, the gap gradually reduced and the bone edges got into a contact. In the fi nal stages, the traces from the contact faded away and the sutural area became a homogenous structure of increased integrity. The μCT scanning of dry bones is a powerful non-destructive technique for examination of the suture morphology. Remodeling of the suture during the obliteration leads to gradually diminishing of the gap between the bone edges to their entire coalescence.

Enzyme histochemical methods are valuable for the studies on the enzyme involvement in different pathological processes. Here we describe two protocols for chromogenic and fluorogenic histochemical demonstration of tripeptidyl aminopeptidase I (TPPI), a protease that is crucial for neuronal functions. The procedures are based on newly synthesized substrates for TPPI-glycyl-L-prolyl-L-metionyl-5-chloro-1-anthraquinonylhydrazide (GPM-CAH) and glycyl-L-prolyl-L-metionyl-4-hydrazido-N-hexyl-1,8-naphthalimide (GPM-HHNI). Using such protocols, precise enzyme localization can be obtained in tissue sections of mammalian organs.

The carotid body (CB) is a small neural crest-derived structure that senses oxygen levels in blood and monitors ventilation. The spontaneously hypertensive rat (SHR) is considered a good experimental model for primary hypertension and is extensively used to study cardiovascular diseases. The hypertensive CB shows structural plasticity and could enlarge without vasodilation. Our immunohistochemical studies revealed the presence of nuclear Ki-67 protein in the sustentacular cells, nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3 and their corresponding receptors p75(NTR), TrkA, TrkB and TrC in the majority of glomus cells and also in a subset of sustentacular cells. In addition, virtually all glomus cells expressed glial cell line-derived neurotrophic factor and its specific receptor GFRα1. The present study demonstrates that in glomus cells of hypertensive animals there is enhanced expression of components of the neurotrophin signaling system compared to normotensive rats. Our results suggest that the elevated production of neurotrophic factors in SHRs could explain CB and sympathetic hyperactivity leading to hypertension.

BACKGROUND A bifid mandibular condyle (BMC) is a rare anatomical variation with an etiology not fully understood. Although there are numerous case reports regarding it, purposeful epidemiological investigations on the BMC frequency among different groups are scarce. This study aims to investigate the incidence and laterality of BMC among series of adult males from Bulgaria and perform a morphometric analysis of it. MATERIALS AND METHODS A series of 500 dry intact mandibles from adult males was investigated. The condyles were macroscopically observed and when skulls were available, the corresponding mandibular fossae were also inspected. In the cases when bifid condyles were found, 27 measurements were taken. RESULTS Bifid mandibular condyle was observed in 4 (0.8%) mandibles. All of the cases were unilateral, 2 on the right side (0.4%) and 2 on the left (0.4%). The condyles were divided into medial and lateral heads by a sagittal fissure or a notch. CONCLUSION The established frequency of BMC was comparable with those reported in Turkish and Korean populations. The lack of injury marks and traces implies a developmental etiology of this condition.

The mesencephalic trigeminal nucleus (Me5) is a unique structure in the central nervous system (CNS), made up of pseudounipolar sensory neurons. It is also a suitable paradigm for studying the plastic alterations in neurons. It is known that the Me5 neurons utilize various neurotransmitters under normal conditions, though little information is available about the morphological and chemical events taking place in the nucleus after injury. This review provides concise description of the structural adaptive changes in Me5 neurons following peripheral axotomy of the masseteric nerve. Furthermore, it validates NADPH-diaphorase activity in them, and using immunohistochemistry for glutamate (Glu), substance P (SP), calcitonin-gene related protein (CGRP), neuropeptide tyrosine (NPY) and galanin (GAL), it deals with the altered neurochemical phenotype of the injured neurons. Our results distinctly show that the Me5 neurons in the rat are extremely sensitive to peripheral injury and we demonstrate their distinct structural and neurochemical plasticity. The adaptive morphological alterations comprise of both qualitative and quantitative alterations in the axotomized Me5 population which are statistically significant when compared with the number and phenotype of the neurons on the contralateral intact side. Besides, the axotomy-induced alterations in the neurochemical character of Me5 are best signified by the down-regulation of the classical neurotransmitters under normal conditions, and the up-regulation of nitric oxide synthase and de novo synthesis of certain neuroactive substances such as NPY, SP, GAL and VIP. It can be inferred that the described phenomena only occur in the nucleus in cases of injury and changes in the environmental cues, and serve as adaptive mechanisms and powerful trophic factors for the neuronal survival in the Me5. There is, undoubtedly, still a long way to go in order to clarify the dynamic and plastic alterations occurring in the CNS in health and disease, and also explain their role in such important functions as pain, perception, learning, cognition and memory.

article i nfo Melatonin is a potent antioxidant which showed anticonvulsant activities both in experimental and clinical studies. In the present study, we examined the effect of melatonin treatment (10 mg/kg/day, diluted in drinking water, 8 weeks) during epileptogenesis on the consequences of a kainate (KA)-induced status epilepticus (SE) in rats. Melatonin increased the latency in the appearance of spontaneous recurrent seizures (SRSs) and decreased their frequency only during the treatment period. The behavioral alterations associated with hyperactivity, depression-like behavior during the light phase, and deficits in hippocampus-dependent working memory were positively affected by melatonin treatment in rats with epilepsy. Melatonin reduced the neuronal damage in the CA1 area of the hippocampus and piriform cortex and recovered the decrease of hippocampal serotonin (5-HT) level in rats with epilepsy. Taken together, long-term melatonin treatment after SE was unable to suppress the development of epileptogenesis. However, it showed a potential in reduc- ing some of the deleterious alterations that develop during the chronic epileptic state in a diurnal phase- dependent mode.