Oguzhan Ekizoglu

Intravenous immunoglobulins prevent the breakdown of the blood-brain barrier in experimentally induced sepsis

Interventions:The effects of immunoglobulin G and immunoglobulins enriched with immunoglobulin A and immunoglobulin M on blood-brain barrier integrity and survival rates in septic rats were comparatively investigated. Measurements:Sepsis was induced by cecal ligation and perforation in Sprague-Dawley rats. The animals were divided into the following groups: Sham, cecal ligation and perforation, cecal ligation and perforation plus immunoglobulin G (250 mg/kg, intravenous), and cecal ligation and perforation plus immunoglobulins enriched with immunoglobulin A and immunoglobulin M (250 mg/kg, intravenous). Immunoglobulins were administered 5 mins before cecal ligation and perforation and the animals were observed for behavioral changes for 24 hrs following cecal ligation and perforation. Blood-brain barrier permeability was functionally and structurally evaluated by determining the extravasation of Evans Blue and horseradish peroxidase tracers, respectively. Immunohistochemistry and Western blotting for occludin were performed. Main Results:The high mortality rate (34%) noted in the septic rats was decreased to 15% and 3% by immunoglobulin G and immunoglobulins enriched with immunoglobulin A and immunoglobulin M, respectively (p < .01). Both immunoglobulin G and immunoglobulins enriched with immunoglobulin A and immunoglobulin M alleviated the symptoms of sickness behavior in the septic rats, with the animals becoming healthy and active. Increased extravasation of Evans Blue into the brain tissue of the septic rats was markedly decreased with the administration of both immunoglobulin G and immunoglobulins enriched with immunoglobulin A and immunoglobulin M (p < .01). Occludin expression remained essentially unchanged in all groups, including the cecal ligation and perforation group. In the cecal ligation and perforation group, increased luminal and abluminal vesicles containing electron-dense horseradish peroxidase-reaction product were noted in the cytoplasm of endothelial cells located in the hippocampus and the cerebral cortex. Tight junction was ultrastructurally intact, suggesting that the transcellular pathway is responsible for the blood-brain barrier breakdown in sepsis. Following immunoglobulin G or immunoglobulins enriched with immunoglobulin A and immunoglobulin M treatment, no ultrastructural evidence of leaky capillaries in the brain was observed in the septic rats, indicating the blockade of the transcellular pathway by immunoglobulins administration. Conclusions:Our study suggests that immunoglobulin G and immunoglobulins enriched with immunoglobulin A and immunoglobulin M improve the integrity of the blood-brain barrier and inhibits cecal ligation and perforation-induced symptoms of sickness behavior in rats. (Crit Care Med 2012; 40:–1220)

Morphological and functional changes of blood–brain barrier in kindled rats with cortical dysplasia

Cortical dysplasia (CD) is one of the major causes contributing to epileptogenesis associated with blood-brain-barrier (BBB) disturbances. The current study investigated the functional and ultrastructural changes of BBB in pentylenetetrazole (PTZ)-kindled rats with CD. Pregnant rats on E17 were exposed to 145 cGy of gamma-irradiation and offspring were used for experiments. The rats were given PTZ three times per week to induce kindling. The permeability of BBB was determined by using sodium fluorescein (NaFlu). Immunohistochemistry for occludin, GFAP and c-fos, western-blot analysis for occludin and electron microscopy for the ultrastructural alterations in BBB were performed. The brain level of NaFlu did not increase in rats with CD and/or kindling. Following administration of a convulsive dose of PTZ, a significant increase in BBB permeability was observed in kindled rats with CD. Occludin immunoreactivity and expression remained essentially unchanged in all groups. Slightly enhanced immunoreactivity for GFAP was observed in all groups except control. c-fos immunoreactivity in brain sections of kindled rats with CD displayed a striking increase by convulsive PTZ challenge. Tight junctions were ultrastructurally intact, whereas markedly increased number of pinocytotic vesicles was noted in brain endothelium of kindled rats with CD by convulsive dose of PTZ. The present study showed that epileptic seizures induced by convulsive PTZ challenge during kindling-mediated epileptogenesis in the presence of CD changed both functional and ultrastructural properties of the BBB and considerably enhanced transendothelial vesicular transport, while paracellular pathway was apparently not involved in this setting.

Levetiracetam decreases the seizure activity and blood-brain barrier permeability in pentylenetetrazole-kindled rats with cortical dysplasia

This study investigates the effects of levetiracetam (LEV) on the functional and structural properties of blood-brain barrier (BBB) in pentylenetetrazole (PTZ)-kindled rats with cortical dysplasia (CD). Pregnant rats were exposed to 145 cGy of gamma-irradiation on embryonic day 17. In offsprings, kindling was induced by giving subconvulsive doses of PTZ three times per week for 45 days. While all kindled rats with CD died during epileptic seizures evoked by the administration of a convulsive dose of PTZ in 15 to 25 min, one week LEV (80 mg/kg) pretreatment decreased the mortality to 38% in the same setting. LEV caused a remarkable decrease (p<0.01) in extravasation of sodium fluorescein dye into the brain tissue of kindled animals with CD treated with convulsive dose of PTZ. Occludin immunoreactivity and expression remained essentially unchanged in all groups. Immunoreactivity for glial fibrillary acidic protein (GFAP) was observed to be slightly increased by acute convulsive challenge in kindled rats with CD while LEV pretreatment led to GFAP immunoreactivity comparable to that of controls. An increased c-fos immunoreactivity in kindled rats with CD exposed to convulsive PTZ challenge was also observed with LEV pretreatment. Tight junctions were ultrastructurally intact, whereas LEV decreased the increased pinocytotic activity in brain endothelium of kindled rats with CD treated with convulsive dose of PTZ. The present study showed that LEV decreased the increased BBB permeability considerably by diminishing vesicular transport in epileptic seizures induced by convulsive PTZ challenge in kindled animals with CD.

Effects of levetiracetam on blood-brain barrier disturbances following hyperthermia-induced seizures in rats with cortical dysplasia

AIMS The mechanisms underlying the changes in blood-brain barrier (BBB) integrity and the generation of seizures in childhood associated with preexisting brain lesions like cortical dysplasia (CD) are poorly understood. We investigated the effects of levetiracetam (LEV) on BBB integrity and the survival during hyperthermic seizures in rats with CD. MAIN METHODS Pregnant rats were exposed to 145 cGy of gamma-irradiation on embryonic day 17. On postnatal day 28, hyperthermia-induced seizures were evoked in offspring with CD. To show the functional and morphological alterations in BBB integrity, quantitative analysis of sodium fluorescein (NaFlu) extravasation, immunohistochemistry and electron microscopy were performed. KEY FINDINGS Seizure scores and mortality rates were decreased by LEV during hyperthermia-induced seizures in rats with CD (P<0.01). Increased NaFlu extravasation into brain by hyperthermia-induced seizures in animals with CD was decreased by LEV (P<0.01). While glial fibrillary acidic protein (GFAP) immunoreactivity slightly increased in brain sections of animals with CD during hyperthermia-induced seizures, LEV led to GFAP immunoreactivity comparable to that of controls. Decreased occludin immunoreactivity and expression in CD plus hyperthermia-induced seizures was increased by LEV. Opening of tight junctions and abundance of pinocytotic vesicles representing ultrastructural evidences of BBB impairment and severe perivascular edema were observed in animals with CD exposed to hyperthermia-induced seizures and LEV treatment led to the attenuation of these findings. SIGNIFICANCE These results indicate that LEV may present a novel approach for the protection of the BBB besides its antiepileptic impact on hyperthermic seizures in the setting of CD.

Forensic age estimation via 3-T magnetic resonance imaging of ossification of the proximal tibial and distal femoral epiphyses: Use of a T2-weighted fast spin-echo technique

Radiation exposure during forensic age estimation is associated with ethical implications. It is important to prevent repetitive radiation exposure when conducting advanced ultrasonography (USG) and magnetic resonance imaging (MRI). The purpose of this study was to investigate the utility of 3.0-T MRI in determining the degree of ossification of the distal femoral and proximal tibial epiphyses in a group of Turkish population. We retrospectively evaluated coronal T2-weighted and turbo spin-echo sequences taken upon MRI of 503 patients (305 males, 198 females; age 10-30 years) using a five-stage method. Intra- and interobserver variations were very low. (Intraobserver reliability was κ=0.919 for the distal femoral epiphysis and κ=0.961 for the proximal tibial epiphysis, and interobserver reliability was κ=0.836 for the distal femoral epiphysis and κ=0.885 for the proximal tibial epiphysis.) Spearmans rank correlation analysis indicated a significant positive relationship between age and the extent of ossification of the distal femoral and proximal tibial epiphyses (p<0.001). Comparison of male and female data revealed significant between-gender differences in the ages at first attainment of stages 2, 3, and 4 ossifications of the distal femoral epiphysis and stage 1 and 4 ossifications of the proximal tibial epiphysis (p<0.05). The earliest ages at which ossification of stages 3, 4, and 5 was evident in the distal femoral epiphysis were 14, 17, and 22 years in males and 13, 16, and 21 years in females, respectively. Proximal tibial epiphysis of stages 3, 4, and 5 ossification was first noted at ages 14, 17, and 18 years in males and 13, 15, and 16 years in females, respectively. MRI of the distal femoral and proximal tibial epiphyses is an alternative, noninvasive, and reliable technique to estimate age.

The use of maxillary sinus dimensions in gender determination: a thin-slice multidetector computed tomography assisted morphometric study.

AbstractGender determination is an important step in identification. For gender determination, anthropometric evaluation is one of the main forensic evaluations. In the present study, morphometric analysis of maxillary sinuses was performed to determine gender. For morphometric analysis, coronal and axial paranasal sinus computed tomography (CT) scan with 1-mm slice thickness was used. For this study, 140 subjects (70 women and 70 men) were enrolled (age ranged between 18 and 63). The size of each subject’s maxillary sinuses was measured in anteroposterior, transverse, cephalocaudal, and volume directions. In each measurement, the size of the maxillary sinus is significantly small in female gender (P < 0.001). When discrimination analysis was performed, the accuracy rate was detected as 80% for women and 74.3% for men with an overall rate of 77.15%. With the use of 1-mm slice thickness CT, morphometric analysis of maxillary sinuses will be helpful for gender determination.

Forensic age estimation by spheno-occipital synchondrosis fusion degree: computed tomography analysis.

Abstract The analysis of ossification points plays a considerable role in forensic age estimation. Although traditional methods are still in use, researchers are working on different age estimation procedures especially within the development of radiologic methods. One of these methods is to define spheno-occipital synchondrosis fusion degree. Spheno-occipital synchondrosis, an important growth point on cranial base, provides noteworthy information about age estimation through its late stage ossification nature. This study aimed to investigate spheno-occipital synchondrosis fusion degree for age estimation in the Turkish population. In our study, 1-mm-sectioned computed tomography images of 638 (399 men and 139 women) subjects within the age of 10 to 25 years were retrospectively examined. It is stated in our study that spheno-occipital syncondrosis fusion begins superiorly and progresses inferiorly until it is completed. Spheno-occipital syncondrosis is known to be totally open at the mean (SD) age of 11.5 (1.5) years in men and 10.7 (0.8) years in women. In addition, fusion degree is known to be increased with age. Fusion starts approximately 2 years earlier in women than in men, and the process of fusion completes at the age of 17 years in both sexes. An analysis of fusion degree between sex groups showed significance at the age of 11 to 15 years, and Spearman rank correlations indicate a significant positive relationship between age and degree of spheno-occipital fusion (P < 0.001; men, &rgr; = 0.714; women, &rgr; = 0.698). Consequently, 5-staged analysis of spheno-occipital synchondrosis fusion degree in use with 1-mm computed tomography images will be helpful for age estimation between 11 and 17 years.

Effects of beta-hydroxybutyrate on brain vascular permeability in rats with traumatic brain injury

This study investigates the effect of beta-hydroxybutyrate (BHB) on blood-brain barrier (BBB) integrity during traumatic brain injury (TBI) in rats. Evans blue (EB) and horseradish peroxidase (HRP) were used as determinants of BBB permeability. Glutathione (GSH) and malondialdehyde (MDA) levels were estimated in the right (injury side) cerebral cortex of animals. The gene expression levels for occludin, glucose transporter (Glut)-1, aquaporin4 (AQP4) and nuclear factor-kappaB (NF-κB) were performed, and Glut-1 and NF-κB activities were analyzed. BHB treatment decreased GSH and MDA levels in intact animals and in those exposed to TBI (P<0.05). Glut-1 protein levels decreased in sham, BHB and TBI plus BHB groups (P<0.05). NF-κB protein levels increased in animals treated with BHB and/or exposed to TBI (P<0.05). The expression levels of occludin and AQP4 did not significantly change among experimental groups. Glut-1 expression levels increased in BHB treated and untreated animals exposed to TBI (P<0.05). While NF-κB expression levels increased in animals in TBI (P<0.01), a decrease was noticed in these animals upon BHB treatment (P<0.01). In animals exposed to TBI, EB extravasation was observed in the ipsilateral cortex regardless of BHB treatment. Ultrastructurally, BHB attenuated but did not prevent the presence of HRP in brain capillary endothelial cells of animals with TBI; moreover, the drug also led to the observation of the tracer when used in intact rats (P<0.01). Altogether, these results showed that BHB not only failed to provide overall protective effects on BBB in TBI but also led to BBB disruption in healthy animals.

Spheno-occipital synchondrosis fusion degree as a method to estimate age: a preliminary, magnetic resonance imaging study

The investigation of ossification regions in the skeletal system is a basic method for forensic age estimation. Although X-ray examination is commonly used in living subjects, magnetic resonance imaging (MRI) has gained importance because of its high resolution and its avoidance of radiation exposure. In this study, we investigated the utility of MRI in the evaluation of Spheno-occipital synchondrosis (SOS) fusion degree as a method to estimate age. The MRI findings of 1078 subjects (455 were male and 623 were female) aged 7–21 years were retrospectively evaluated according to a five-stage method. We found that SOS fusion started two years earlier in female subjects. The average age for stage 5 SOS fusion was 18.43 ± 1.84 years in male subjects and 17.78 ± 2.20 years in female subjects. Our results show that MRI in the determination of SOS fusion degree is an efficient non-invasive method to estimate age and avoids the disadvantages of radiation exposure incurred by X-ray or computed tomography examinations.

Virtual Assessment of Sex: Linear and Angular Traits of the Mandibular Ramus Using Three-Dimensional Computed Tomography.

AbstractMorphometric analysis of the mandibular ramus (MR) provides highly accurate data to discriminate sex. The objective of this study was to demonstrate the utility and accuracy of MR morphometric analysis for sex identification in a Turkish population.Four hundred fifteen Turkish patients (18–60 y; 201 male and 214 female) who had previously had multidetector computed tomography scans of the cranium were included in the study. Multidetector computed tomography images were obtained using three-dimensional reconstructions and a volume-rendering technique, and 8 linear and 3 angular values were measured. Univariate, bivariate, and multivariate discriminant analyses were performed, and the accuracy rates for determining sex were calculated.Mandibular ramus values produced high accuracy rates of 51% to 95.6%. Upper ramus vertical height had the highest rate at 95.6%, and bivariate analysis showed 89.7% to 98.6% accuracy rates with the highest ratios of mandibular flexure upper border and maximum ramus breadth. Stepwise discrimination analysis gave a 99% accuracy rate for all MR variables.Our study showed that the MR, in particular morphometric measures of the upper part of the ramus, can provide valuable data to determine sex in a Turkish population. The method combines both anthropological and radiologic studies.