5th Postnatal Day Research Articles

Influence of Prenatal Hypoxia on the Content of Neuron Specific Enolase in the Structures of the Brain and Blood Serum of Rats in Early Ontogeny

Abstract—We studied the dynamics of changes in the content of neuron specific enolase (NSE, EC 4.2.1.11) in the hippocampus, cortex, cerebellum and blood serum of rats at 5th, 10th, and 30th days of postnatal development. It was shown that during the first month of life the content of NSE increases several-fold in all the structures studied, both after hypoxia and in the animals not exposed to hypoxia. It was established that prenatal hypoxia on the 14th day of embryonic development leads to significant changes in the NSE content in all the brain structures studied. In the blood serum, the content of this enzyme is higher in comparison with the control group on the 5th, 10th, and 30th postnatal days in experimental animals. Thus, the lack of oxygen affects the content of this enzyme in both the brain structures and in blood serum of rats, which makes it possible to judge the survival of neurons under conditions of prenatal hypoxia, and also suggests that this index may be used as a marker of fetal CNS disturbance after hypoxia during prenatal development.

Abstract 51: The Incidence of Perinatal Stroke is 1:1200 Live Births: A Population-Based Study in Alberta Canada

Introduction: Perinatal stroke encompasses six cerebrovascular syndromes which occur between the 20 th week of gestation and the 28 th post-natal day. Morbidity is significant including motor, language, behavioral, and cognitive challenges, as well as epilepsy. Acute presentations include neonatal arterial ischemic stroke (NAIS), neonatal cerebral sinovenous thrombosis (CSVT), and neonatal hemorrhagic stroke (NHS). Delayed presentations include arterial presumed perinatal ischemic stroke (APPIS), periventricular venous infarction (PVI), and presumed perinatal hemorrhagic stroke (PPHS). Inconsistent terminology and lack of population-based cohorts has limited accurate measurement of disease-specific perinatal stroke incidence. Our objective was to define the incidence of the subtypes of perinatal stroke using a population-based cohort. Methods: The Alberta Perinatal Stroke Project is a research cohort established in 2008 in Southern Alberta, Canada (population ~2.1 million). Leveraging universal health care at a single tertiary care pediatric center facilitated true population-based epidemiology. Patients included had neuroimaging-confirmed perinatal stroke. Case acquisition included exhaustive retrospective hospital and ICD code searches (1990-2008) and prospective enrollment from all NICU and neurology/stroke clinics (2008-2018). Live birth rate denominators were determined from the provincial census. Results: The overall incidence of perinatal stroke in Southern Alberta from 2008-2018 was 81 cases per 100,000 live births, or 1 case per approximately 1200 live births. The incidence of NAIS was 31 per 100,000 (~1/3000), APPIS was 11 per 100,000 (~1/9000), PVI was 18 per 100,000 (~1/5500), and CSVT was 10 per 100,000 (~1/9800). The incidence of NAIS increased after implementing prospective case identification, from 8.5 (95% CI 5.3-11.6) per 100,000 to 31 per 100,000 (95% CI 24.4-38.2), p<0.00001, the remainder of stroke subtypes were stable over time. Conclusions: The overall incidence of perinatal stroke in Southern Alberta is 1:1200 live births. Population-based sampling of disease-specific states may explain why this rate is much higher than previous estimates.

Влияние раннего провоспалительного стресса на тревожное и депрессивно-подобное поведение крыс разного возраста

An early proinflammatory stress produced by administration of LPS (50 mcg/kg, sc) on 3rd and 5th postnatal days led to decrease of locomotor and search activity and increase of anxiety in rats at the age of 1 month in the tests of an open field and elevated plus maze. The males showed more anxious behavior than females. Administration of LPS in early ontogenesis produced the signs of depressive-like behavior in males and females in the test of anhedonia by decrease of preference of 1% sucrose solution in comparison with control groups. The forced swimming test showed the signs of depressive-like behavior in males, but not in females at the first day of testing at the age of 1 month. At the age of 3 months the all mentioned changes in anxious and depressive-like behavior were practically disappeared. The data obtained point out to the sex differences in the effects of early proinflammatory stress on anxious and depressive-like behavior in rats.

LPS-Induced Systemic Neonatal Inflammation: Blockage of P2X7R by BBG Decreases Mortality on Rat Pups and Oxidative Stress in Hippocampus of Adult Rats.

Neonatal lipopolysaccharide (LPS) exposure-induced brain inflammation has been associated to neuronal injury and facilitates the development of models of neurological disorders in adult rats. The P2X7 receptor (P2X7R) plays a fundamental role in the onset and maintenance of the inflammatory cascade. Brilliant blue G (BBG), a P2X7R antagonist, has been shown to effectively promote neuroinflammatory protection. Here, we have investigated the long-term effects of the neonatal systemic inflammation on hippocampal oxidative stress, anxiety behavior and pain sensitivity in adulthood. We hypothesized that P2X7R blockade is able to modulate the effects of inflammation on these variables. Male and female rat pups received LPS and/or BBG solution intraperitoneally on the 1st, 3rd, 5th and 7th postnatal days. The survival rate and body weight were evaluated during the experimental procedures. The animals were submitted to behavioral tests for anxiety (elevated plus maze, EPM) and nociception (hot-plate and tail-flick) and the oxidative stress was measured by superoxide production in the dentate gyrus of the hippocampus using dihydroethidium (DHE) probe. BBG increased the survival rate in LPS-treated rats. No significant differences were found regarding anxiety behavior and pain sensitivity between the experimental groups. Systemic neonatal inflammation leads to a higher production of superoxide anion in the dentate gyrus of the hippocampus in adulthood and BBG inhibited that effect. Our data suggest that blocking the activation of the P2X7R during neonatal systemic inflammation may have a potential neuroprotective effect in adulthood.

The effects of bosentan on hyperoxia-induced lung injury in neonatal rats.

Bronchopulmonary dysplasia (BPD) remains an important cause of morbidity and mortality in premature infants. There is currently no proven effective treatment modality for BPD, and inflammation and oxidative injury play an important role in the pathogenesis of this disease. This study investigated the histopathological and biochemical effects of bosentan, which is a non-specific endothelin receptor antagonist with known antioxidant and anti-inflammatory properties, on hyperoxia-induced lung injury (HILI) in neonatal rats. The experiment was performed on newborn rats from the 3rd to the 13th postnatal day. The rats were randomly divided into six groups: Group 1 (air-exposed+saline, n=6); Group 2 (HILI, n=8); Group 3 (air-exposed+bosentan, n=7); Group 4 (HILI+saline, n=7); Group 5 (HILI+early bosentan-treated group, n=6), and Group 6 (HILI+late bosentan-treated group, n=7). Bosentan was administered (30mg/kg/day) intraperitoneally. The histopathological effects of bosentan on lung tissue were assessed by their alveolar surface area, fibrosis, and smooth muscle actin (SMA) scores, and the biochemical effects on lung tissue were assessed by interleukin-1 beta (IL-1β), IL-6, IL-10, and tumor necrosis factor-alpha (TNF-α). The alveolar surface area and fibrosis scores were found to be significantly higher in HILI groups compared with Group 1 (P<0.01). The SMA scores in HILI groups were also significantly higher than Group 1 (P<0.01). Bosentan treatment, especially late therapy, reduced all of these histopathological scores and the levels of IL-6 and TNF-α in the hyperoxia groups (P<0.01). This experimental study showed that bosentan had a protective effect on hyperoxic lung injury through its anti-inflammatory properties.

Valproic acid concentrations in nursing mothers, mature milk, and breastfed infants in monotherapy and combination therapy

Valproic acid (VPA) is currently one of the four most often prescribed antiepileptic drugs (AEDs) in pregnancy. However, only a small number of studies have measured suckling infant serum levels of the drug. We studied the transport of VPA from breastfeeding mothers to the mature milk and breastfed infants and the influence of comedication with enzyme-inducing AEDs. The data of 30 nursing women treated by VPA were analyzed retrospectively. Mature milk, maternal, and infant serum levels were collected between the 6th and 32nd postnatal day and measured by gas chromatography during the years 1996–2017.Valproic acid levels varied from 5.4 to 69.0 mg/L (mean: 39.0 ± 16.1 mg/L) in the maternal serum, from <1.0 to 16.7 mg/L (mean: 1.6 ± 3.9 mg/L) in the milk, and from <1.0 to 17.5 mg/L (mean: 4.2 ± 4.3 mg/L) in the infant serum. The milk/maternal serum level ratio ranged from <0.03 to 0.25 (mean: 0.03 ± 0.06) and the infant/maternal serum level ratio from <0.03 to 0.61 (mean: 0.11 ± 0.13). Sixty-seven percent of milk and 33% of infant VPA concentrations were below the limit of quantification. No correlations were observed between maternal serum and milk levels or between maternal and infant serum levels. In conclusion, none of the milk or infant serum VPA levels reached the lower limit of the reference range used for the general population with epilepsy, so the degree of VPA exposure in breastfed infants is less than during gestation. Nevertheless, if signs of potential adverse reactions manifest, infant serum concentrations should be measured.

Giant Congenital Intracranial Immature Teratoma: A Case Report and Review of Literature

AbstractCongenital intracranial tumour constitute only 0.5–1.5% and are usually associated with poor prognosis. New advancement in imaging may help in early diagnosis. We presented a case of giant fetal intracranial tumour diagnosed antenatally at 28 weeks of gestation in sonography. The magnetic resonance imaging confirmed a large hetrogenous intracranial mass with bilateral ventriculomegaly. Because of the large size of the tumor and associated macrocephaly, the baby was delivered by caessaran section. On delivery, fetal head size was 44.5 cm with wide suture and left facial nerve was involved. Ventriculoeritoneal shunt was inserted on 4th postnatal day, however complete surgical resection of tumour was not performed because of parents refusal. Neonate had rapid increase in head size and expired on postnatal day 16th. The histopathological findings of mass demonstrated a congenital immature teratoma. Neonate expired on 16th postnatal day. Antenatal diagnosis of intracranial tumour is possible with advanced imaging modalities, however an early diagnosis before second trimester is difficult. Prognosis is generally poor and depends on size of tumour, histology and timely intervention.

Hyperactivity and memory/learning deficits evoked by developmental exposure to nicotine and/or ethanol are mitigated by cAMP and cGMP signaling cascades activation

Pregnant smoking women are frequently episodic drinkers. Here, we investigated whether ethanol exposure restricted to the brain growth spurt period when combined with chronic developmental exposure to nicotine aggravates memory/learning deficits and hyperactivity, and associated cAMP and cGMP signaling disruption. To further investigate the role of these signaling cascades, we verified whether vinpocetine (a phosphodiesterase inhibitor) ameliorates the neurochemical and behavioral outcomes. Swiss mice had free access to nicotine (NIC, 50 μg/ml) or water to drink during gestation and until the 8th postnatal day (PN8). Ethanol (ETOH, 5 g/kg, i.p.) or saline were injected in the pups every other day from PN2 to PN8. At PN30, animals either received vinpocetine (20 mg/kg, i.p.) or vehicle before being tested in the step-down passive avoidance or open field. Memory/learning was impaired in NIC, ETOH and NIC + ETOH mice, and vinpocetine mitigated ETOH- and NIC + ETOH-induced deficits. Locomotor hyperactivity identified in ETOH and NIC + ETOH mice was ameliorated by vinpocetine. While cyclic nucleotides levels in cerebral cortex and hippocampus were reduced by NIC, ETOH and NIC + ETOH, this outcome was more consistent in the latter group. As observed for behavior, vinpocetine normalized NIC + ETOH nucleotides levels. pCREB levels were also increased in response to vinpocetine, with stronger effects in the NIC + ETOH group. Exposure to both drugs of abuse worsens behavioral and neurochemical disruption. These findings and the amelioration of deleterious effects by vinpocetine support the idea that cAMP and cGMP signaling contribute to nicotine- and ethanol-induced hyperactivity and memory/learning deficits.

Analysis of Housekeeping Genes for Accurate Normalization of qPCR Data During Early Postnatal Brain Development.

Maturation of the neocortex during the first three postnatal weeks is a complex process that is characterized by different time courses of maturation of different areas of the neocortex. Analysis of gene expression using quantitative PCR after reverse transcription during this period of ontogeny and comparison of different cortical areas require optimal selection of reference genes for correct normalization of the data. Here, we compared expression of nine reference genes in the somatosensory and visual areas of the neocortex at the age of 5, 8, 10, 13, and 20days. Using widely used GeNorm and NormFinder applications, as well as a novel approach, we compared stability of expression of GAPDH, YWHAZ, TFR1, RPS18, Rn18S, HPRT1, KIF5C, OSBP, and UQCRFS1. We found that, in both neocortical areas studied, YWHAZ and UQCRFS1 are the best reference genes whereas GAPDH and TFR1 are also stably expressed in the somatosensory cortex and OSBP is stable in the visual cortex. Additionally, analysis of stability of expression of these genes by a novel approach showed that the expression of these genes is stable during the entire period from the 5th to the 20th postnatal days.

Haloperidol affects bones while clozapine alters metabolic parameters - sex specific effects in rats perinatally treated with phencyclidine

BackgroundThe presentation of schizophrenia (SCH) symptoms differs between the sexes. Long-term treatment with antipsychotics is frequently associated with decreased bone mineral density, increased fracture risk and metabolic side effects. Perinatal phencyclidine (PCP) administration to rodents represents an animal model of SCH. The aim of this study was to assess the effects of chronic haloperidol and clozapine treatment on bone mass, body composition, corticosterone, IL-6 and TNF-α concentrations and metabolic parameters in male and female rats perinatally treated with PCP.MethodsSix groups of male and six groups of female rats (n = 6-12 per group) were subcutaneously treated on 2nd, 6th, 9th and 12th postnatal day (PN), with either PCP (10 mg/kg) or saline. At PN35, one NaCl and PCP group (NaCl-H and PCP-H) started receiving haloperidol (1 mg/kg/day) and one NaCl and PCP group (NaCl-C and PCP-C) started receiving clozapine (20 mg/kg/day) dissolved in drinking water. The remaining NaCl and PCP groups received water. Dual X-ray absorptiometry measurements were performed on PN60 and PN98. Animals were sacrificed on PN100. Femur was analysed by light microscopy. Concentrations of corticosterone, TNF-α and IL-6 were measured in serum samples using enzyme-linked immunosorbent assay (ELISA) commercially available kits. Glucose, cholesterol and triacylglycerol concentrations were measured in serum spectrophotometrically.ResultsOur results showed that perinatal PCP administration causes a significant decrease in bone mass and deterioration in bone quality in male and female rats. Haloperidol had deleterious, while clozapine had protective effect on bones. The effects of haloperidol on bones were more pronounced in male rats. It seems that the observed changes are not the consequence of the alterations of corticosterone, IL-6 and TNF-α concentration since no change of these factors was observed. Clozapine induced increase of body weight and retroperitoneal fat in male rats regardless of perinatal treatment. Furthermore, clozapine treatment caused sex specific increase in pro-inflammatory cytokines.ConclusionTaken together our findings confirm that antipsychotics have complex influence on bone and metabolism. Evaluation of potential markers for individual risk of antipsychotics induced adverse effects could be valuable for improvement of therapy of this life-long lasting disease.

Effect of Sitz Bath on Episiotomy Wound Healing and Level of Pain among Post Natal Mothers

Enhancing healing of episiotomy wound is one of the major concerns after a normal delivery. The study aimed to identify the effectiveness of sitz bath on reducing pain and wound healing. With use of universal pain scale and REEDA scale were measured from sample of 20 from experimental and control group on 1st, 3rd and 5th postnatal day. The experimental group had a significant improvement in wound healing and level of pain.

Effects of Maternal Hyperhomocysteinemia on the Early Physical Development and Neurobehavioral Maturation of Rat Offspring

During pregnancy, several complications have been associated with hyperhomocysteinemia (HHcy) and elevated homocysteine (Hcy) levels have been shown to play a role in the etiology of preeclampsia, placental abruption, intrauterine growth retardation, and neural tube defects and associated with the neurological consequences. In the present work, we investigated the effects of chronic maternal HHcy on the development and neurobehavioral maturation of the offspring. We analyzed classical parameters of development such as body weight, eyelid opening, ear unfolding, incisor eruption, and the appearance of hair, and subjected the pups to various tests that reflected the neurobehavioral maturation extending from 4th to 20th postnatal days (righting reflex, negative geotaxis, cliff avoidance, head shake, acoustic startle reflex, free-fall righting, cliff avoidance caused by visual stimulus, olfactory discrimination). We have shown that newborn animals were characterized by lower body weight and higher mortality. Besides, the delay in neurobehavioral maturation of the pups from the Hcy group was observed. The obtained results indicate that early developmental impairments of brain maturation induced by prenatal HHcy may underlie long-term deficits in the learning and memory behaviors.

Impact of targeted-volume ventilation on pulmonary dynamics in preterm infants with respiratory distress syndrome.

Mechanical ventilation is an essential therapy in the treatment of respiratory failure in preterm infants. However, optimal ventilation strategy continues to be difficult to define. To compare the effects of volume guarantee (VG) combined with intermittent mandatory ventilation (SIMV) and VG combined with pressure support ventilation (PSV) on the pulmonary mechanics and short term prognosis in preterm infants with respiratory distress syndrome. Infants of <32 weeks gestational age ventilated for respiratory distress syndrome were randomized to receive either SIMV + VG or PSV + VG. The patient characteristics, ventilator variables including PIP, PEEP, MAP, VT, dynamic compliance, resistance, C20/C, and neonatal outcomes (IVH, ROP, oxygen dependency at 28th postnatal day and 36 weeks of PMA), mortality and extubation failure were recorded in each groups. Thirty-four infants were enrolled in to the study: 19 patients were randomized to the SIMV + VG group, and 15 patients to the PSV + VG group. No significant differences were observed between the two groups in terms of the birth weight, gestational age, gender, multiple pregnancy, delivery mode, and antenatal steroid treatment. The respiratory and ventilatory parameters were similar in the groups. The need for reintubation were common in SIMV + VG group (P < 0.01). Volume guaranteed ventilation combined with PSV may be a convenient method for preterm infants with RDS in terms of reducing postextubation atelectasis and the need for reintubation. Pediatr Pulmonol. 2017;52:213-216. © 2016 Wiley Periodicals, Inc.

Effect of creatine monohydrate supplementation on learning, memory and neuromuscular coordination in female albino mice.

Research findings made over the last few years have highlighted the important role of creatine (Cr) in health and disease. However, limited information is available regarding the effect of Cr supplementation on cognation. Present study was designed to determine the effect of variable doses of Cr (1% and 3%) on selected parameters of female albino mice behaviour. Following weaning, on 20th postnatal day, female albino mice were divided into three groups on the basis of dietary supplementation. Control group were was fed with normal rodent diet, whereas treated groups received diet supplemented with 1% and 3% Creatine monohydrate (Ssniff, Germany) for 10 weeks. Morris water maze (MWM), Rota rod and open field (OF) tests were carried out at the end of diet supplementation for neurofunctional assessment in all the groups. Data analysis showed that Cr supplementation did not affect the muscular activity and during rota rod test as well as locomotor and exploratory behaviour during OF test. Results of MWM probe trial indicated that mice supplemented with 3% Cr had significantly more entries in platform area than other two treatments (p=0.03) indicating improved spatial memory. Body weight remained unaffected (p>0.05) when compared between three experimental treatments. Female mice supplemented with 3% Cr showed improved spatial memory than mice fed on 1% Cr-supplemented diet and mice on normal rodent diet.

Effects of maternal low-protein diet on parameters of locomotor activity in a rat model of cerebral palsy

Children with cerebral palsy have feeding difficulties that can contribute to undernutrition. The aim of this study was to investigate the effect of early undernutrition on locomotor activity and the expression of the myofibrillar protein MuRF-1 in an experimental model of cerebral palsy (CP). In order to achieve this aim, pregnant rats were divided into two groups according to the diet provided: Normal Protein (NP, n=9) and Low Protein (LP, n=12) groups. After birth, the pups were divided into four groups: Normal Protein Sham (NPS, n=16), Normal Protein Cerebral Palsy (NPCP, n=21), Low Protein Sham (LPS, n=20) and Low Protein Cerebral Palsy (LPCP, n=18) groups. The experimental cerebral palsy protocol consisted of two episodes of anoxia at birth and during the first days of life. Each day, nitrogen flow was used (9l/min during 12min). After nitrogen exposure, sensorimotor restriction was performed 16h per day, from the 2nd to the 28th postnatal day (PND). Locomotor activity was evaluated at 8th, 14th, 17th, 21th and 28th PND. At PND 29, soleus muscles were collected to analyse myofibrillar protein MuRF-1. Our results show that CP animals decreased body weight (p<0.001), which were associated with alterations of various parameters of locomotor activity (p<0.05), compared to their control. Undernourished animals also showed a decrease (p<0.05) in body weight and locomotor activity parameters. Moreover, CP decreased MuRF-1 levels in nourished rats (p=0.015) but not in undernourished rats. In summary, perinatal undernutrition exacerbated the negative effects of cerebral palsy on locomotor activity and muscle atrophy, but it appears not be mediated by changes in MuRF-1 levels.

Role of colonic mast cell infiltration and nerve growth factor in visceral hypersensitivity of irritable bowel syndrome

Objective To investigate the role of mucosal mast cells infiltration and degranulation with nerve growth factor (NGF) in development of visceral hypersensitivity in Sprague-Dawley (SD) rats. Methods The model of visceral hypersensitivity of irritable bowel syndrome (IBS) was established in 19 neonate SD rats with intestinal stimulation (rectalballon distention) on 8th, 10th and 12th postnatal days. The other 19 neonate SD rats without colonic distention were assigned to the control group. After rats grew up (six to eight weeks old), the visceral sensitivity was tested by abdominal withdrawal reflex (AWR) in 10 rats of each group. Mast cell infiltration and degranulation were observed with toluidine blue staining in colon tissue slides. The NGF level of intestinal tissues was detected by enzyme-linked immunosorbent assay (ELISA) methods in the left nine rats of each group. The culture system of dorsal root ganglias (DRG) from the neonatal rats was set up. The changes of electrophysilogical characters of DRG stimulated with NGF (100 ng/mL) for four days were recorded with patch-clamp.Paired t test was performed for comparison between groups. Results The results of AWR indicated that neonatal colonic stimulation could significantly increase visceral sensitivity after growing up. Under 20, 40 and 60 mmHg (1 mmHg=0.133 kPa) distention pressure, visceral sensitivity scores of visceral hypersensitivity rats and rats of control group were 1.00±0.50 vs 1.67±0.50, 1.89±0.31 vs 2.89±0.34 and 2.89±0.33 vs 3.89±0.33, the differences were statistically significant (t=-2.83, -6.00 and -6.00, all P<0.05). The results of master cells staining in tissue slides showed colonic master cells infiltration was obvious in rats with visceral hypersensitivity, and part of mast cells were degranulation. The result of ELISA demonstrated that NGF level of visceral hypersensitivity rats was significantly higher than that of control group ((11.07±3.06) pg/mg vs (2.38±1.88) pg/mg, t=-6.93, P<0.05). The results of electrophysilogical tests of primary cultured DRG indicated that compared with blank control growp, the action potential threshold of neuron in NGF 100 ng/mL group significantly decreased ((-18.0±2.1) mV vs (-29.0±2.5) mV, t=12.26, P<0.05) and discharge frequency increased ((5.0±1.4)/800 ms vs (12.0±3.2)/800 ms, t=-8.40, P<0.05). Meanwhile, neuron voltage-gated K+ current density remarkably decreased, most were sustained delayed rectifier K+ current (IK) decreasing ((279.0±48.0) pA/pF vs (203.0±39.0) pA/pF, t=6.18, P<0.05). Conclusion Colonic stimulation in neonatal rats could cause intestinal master cells infiltration and degranulation, which induced changes of neuron electrophysilogical characters and resulted in visceral hypersensitivity after growing up. Key words: Irritable bowel syndrome; Hyperesthesia; Ganglion, spinal; Mast cells; Nerve growth factor

Mitochondrial impairment, apoptosis and autophagy in a rat brain as immediate and long-term effects of perinatal phencyclidine treatment — influence of restraint stress

Phencyclidine (PCP) acts as a non-competitive antagonist of glutamatergic N-methyl-d-aspartate receptor. Its perinatal administration to rats causes pathophysiological changes that mimick some pathological features of schizophrenia (SCH). Numerous data indicate that abnormalities in mitochondrial structure and function could be associated with the development of SCH. Mitochondrial dysfunction could result in the activation of apoptosis and/or autophagy. The aim of this study was to assess immediate and long-term effects of perinatal PCP administration and acute restraint stress on the activity of respiratory chain enzymes, expression of apoptosis and autophagy markers and ultrastructural changes in the cortex and hippocampus of the rat brain.Six groups of rats were subcutaneously treated on 2nd, 6th, 9th and 12th postnatal days (P), with either PCP (10mg/kg) or saline (0.9% NaCl). One NaCl and one PCP group were sacrificed on P13, while other two NaCl and PCP groups were sacrificed on P70. The remaining two NaCl and PCP groups were subjected to 1h restraint stress prior sacrifice on P70. Activities of respiratory chain enzymes were assessed spectrophotometrically. Expression of caspase 3 and AIF as markers of apoptosis and Beclin 1, p62 and LC3, as autophagy markers, was assessed by Western blot. Morphological changes of cortical and hippocampal ultrastructure were determined by transmission electron microscopy.Immediate effects of perinatal PCP administration at P13 were increased activities of complex I in the hippocampus and cytochrome c oxidase (COX) in the cortex and hippocampus implying mitochondrial dysfunction. These changes were followed by increased expression of apoptotic markers. However the measurement of autophagy markers at this time point has revealed decrease of this process in cortex and the absence of changes in hippocampus. At P70 the activity of complex I was unchanged while COX activity was significantly decreased in cortex and increased in the hippocampus. Expressions of apoptotic markers were still significantly higher in PCP perinatally treated rats in all investigated structures, but the changes of autophagy markers have indicated increased level of autophagy also in both structures. Restraint stress on P70 has caused increase of COX activity both in NaCl and PCP perinatally treated rats, but this increase was lower in PCP group. Also, restraint stress resulted in decrease of apoptotic and increase of autophagy processes especially in the hippocampus of PCP perinatally treated group. The presence of apoptosis and autophagy in the brain was confirmed by transmission electron microscopy.In this study we have demonstrated for the first time the presence of autophagy in PCP model of SCH. Also, we have shown increased sensitivity of PCP perinatally treated rats to restraint stress, manifested in alterations of apoptotic and autophagy markers. The future studies are necessary to elucidate the role of mitochondria in the pathophysiology of SCH and putative significance for development of novel therapeutic strategies.

Inflammation induced at different developmental stages affects differently the range of microglial reactivity and the course of seizures evoked in the adult rat.

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. 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. In naïve 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. 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".

A clinical scoring system to predict the development of bronchopulmonary dysplasia.

This study aims to develop a scoring system for the prediction of bronchopulmonary dysplasia (BPD). Medical records of 652 infants whose gestational age and birth weight were below 32 weeks and 1,500 g, respectively, and who survived beyond 28th postnatal day were reviewed retrospectively. Logistic regression methods were used to determine the clinical and demographic risk factors within the first 72 hours of life associated with BPD, as well as the weights of these factors on developing BPD. Predictive accuracy of the scoring system was tested prospectively at the same unit. Birth weight, gestational age, gender, presence of respiratory distress syndrome, patent ductus arteriosus, intraventricular hemorrhage, hypotension were the most important risk factors for BPD. Therefore, a scoring system (BPD-TM score) ranging from 0 to 13 and grouped in four tiers (0-3: low, 4-6: low intermediate, 7-9: high intermediate, and 10-13: high risk) was developed based on these factors. Below the score of 4, 4.1% of infants (18/436), above the score of 9, 100% (29/29) of the infants developed BPD. The score was validated successfully in 172 infants. With this easy to use scoring system, one can predict the neonate at risk for BPD at 72 hours of life and direct preventive measures toward these infants.

PO-0716 Continuous Venovenous Hemodiafiltration Experience Of Four Newborns

Aim Conventional methods are the first treatment modalities of renal failure or metabolic diseases in newborns. If these modalities fail to treat, we start to use peritoneal dialysis (PD). Continuous venovenous hemodiafiltration (vvHDF) is used when PD can not be performed. Our continuous vvHDF experience of 4 patients in neonatal intensive care unit, is presented. Case1: A male term newborn infant, having mapple syrup urine disease with a high serum leucine value after PD could not be performed, vvHDF was successfully provided. He was discharged from our hospital on 34th postnatal day. Case2: A preterm newborn, having polycystic renal disease and could not use under PD and vvHDF was started on 13th postnatal day. He died due to ventilator associated pneumonia on 135th postnatal day. Case3: A term newborn, having “polycystic renal disease” and could not perform PD, was referred to our unit for continuous vvHDF administration on 3rd postnatal day. vvHDF application was continued until his 61st postnatal day. Case4: A term newborn was appealed to our emergency service, with dyspnea and supraventricular tachycardia was diagnosed on 13th postnatal day. After intervention, multiorgan failure developed in our patient. At his postnatal day 27, vvHDF was performed. The patient died because of ventilator associated pneumonia. Conclusion Continuous vvHDF application should be considered in the neonatal period, in cases where it is impossible to apply PD. Due to the technical difficulties in the neonatal period, such application is not common but it is also life saving.