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Background: Neonatal white matter injury (NWMI) is the leading cause of cerebral palsy in prematurely born children. In order to develop a test bed for therapeutics, we recently reported a mouse model of NWMI by using a modified Rice-Vannucci model of neonatal ischemia on postnatal day 5 (P5) in CD-1 mice. We have previously shown that these mice illustrate initial neuroinflammation and oligodendroglial differentiation arrest followed by long-term dysmyelination, periventricular astrogliosis and axonal injury, resembling human NWMI. The objective of this study was to determine the sex-dependent long-term effects of neonatal brain injury on neurobehavioral and advanced in vivo neuroimaging indices in this mouse model, and to correlate these variables with histopathology., Methods: After right common artery ligation on P5, in vivo T2-weighted imaging and diffusion tensor imaging (DTI) were performed on ligated and control animals at 4 and 8 weeks. Common sets of regions of interest were used to compare fractional anisotropy (FA) values between ischemic and control mice. Behavioral testing (open field, startle response and grip strength) was performed at adult age. Finally, the animals were sacrificed for immunohistochemical (IHC) assessment of major white matter tracts., Results: DTI revealed significant sex-dependent changes in FA values ipsi- and contralateral to the ligation. Behavioral testing showed decreased reaction to acoustic stimuli in males but not females. Similarly, increased number of rearings and lack of novelty-induced habituation in the open field were encountered only in the male subgroup. Several regional correlations were found between FA values and these behavioral alterations. IHC studies revealed degeneration of mature oligodendrocytes and damage of white matter tracts in ligated animals, as previously reported in this model, and showed regional correlation with in vivo FA values and behavioral alterations., Conclusions: Our findings suggest structural sex-dependent long-term abnormalities after neonatal ischemia. These changes lead to behavioral deficits resembling common problems of patients with cerebral palsy., (© 2016 S. Karger AG, Basel.)

Objective The authors sought to assess the relationship between low oxygen delivery (DO 2) during cardiopulmonary bypass (CPB) and a neuron-specific biomarker of neurologic injury, ubiquitin C-terminal hydrolase L1 (UCH-L1). Design Retrospective analysis of patient charts and prospectively collected blood samples. Setting University-affiliated tertiary care hospital. Participants Adult patients undergoing cardiac surgery on CPB. Interventions Serum UCH-L1 levels were drawn at baseline and 6 and 24 hours after CPB cessation. DO 2 was computed from perfusion records, with area-under-the-curve (AUC) computations performed to account for distance of DO 2 excursions below predefined DO 2 thresholds and the amount of time spent below them. Strokes were defined radiographically using computed tomography and magnetic resonance imaging. Measurements and Main Results Forty-three adults were included (median age 65 y, interquartile range 59-72). Three patients experienced strokes (imaged at 2, 7, and 8 d postoperatively). Most patients underwent isolated coronary artery bypass grafting (41%, 18 patients) or isolated aortic valve replacement (30%, 13). Median UCH-L1 levels differed from baseline to 6 and 24 hours after CPB (40, 232, and 166 pg/mL, respectively; p < 0.001). On multivariable linear regression analysis controlling for baseline and surgical variables, only DO 2 AUC <225 was significantly associated with 6-hour UCH-L1 levels (p = 0.001), whereas only DO 2 AUC <300 was significantly associated with 24- hour levels (p < 0.001). The 3 patients who experienced radiographic strokes had nonsignificantly elevated 24-hour UCH-L1 levels compared with control patients (585 v 151 pg/mL, p = 0.11). Conclusions This is the first study to demonstrate an independent association between DO 2 during CPB and elevations of a brain injury biomarker; additional study is needed to clarify the clinical significance of these results. [ABSTRACT FROM AUTHOR]

Microglial activation in crossing white matter tracts is a hallmark of noncystic periventricular leukomalacia (PVL), the leading pathology underlying cerebral palsy in prematurely born infants. Recent studies indicate that neuroinflammation within an early time window can produce long-lasting defects in oligodendroglial maturation, myelination deficit, as well as disruption of transcription factors important in oligodendroglial maturation. We recently reported an ischemic mouse model of PVL, induced by unilateral neonatal carotid artery ligation, leading to selective long-lasting bilateral myelination deficits, ipsilateral thinning of the corpus callosum, ventriculomegaly, as well as evidence of axonopathy. Here, we report that permanent unilateral carotid ligation on postnatal day 5 in CD-1 mice induces an inflammatory response, as defined by microglial activation and recruitment, as well as significant changes in cytokine expression (increased IL-1β, IL-6, TGF-β1, and TNF-α) following ischemia. Transient reduction in counts of oligodendrocyte progenitor cells (OPCs) at 24 and 48 h after ischemia, a shift in OPC cell size and morphology towards the more immature form, as well as likely migration of OPCs were found. These OPC changes were topographically associated with areas showing microglial activation, and OPC counts negatively correlated with increased microglial staining. The presented data show a striking neuroinflammatory response in an ischemia-induced model of PVL, associated with oligodendroglial injury. Future studies modulating the neuroinflammatory response in this model may contribute to a better understanding of the interaction between microglia and OPCs in PVL and open opportunities for future therapies. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Abstract: Stroke is an important cause of neurologic injury in the neonatal period and frequently results in lifelong neurologic impairments. We reported previously that unilateral carotid ligation on postnatal day (P)12 in CD1 mice causes acute behavioral seizures and unilateral brain injury and provides a model for neonatal stroke in human infants. In the present study we confirmed that behavioral seizures observed after ligation on P12 in the CD1 strain are associated with rhythmic ictal discharges that show temporal progression on electrocorticograms. We also examined the effects of carotid ligation performed at different ages in CD1 mice or performed in the C57Bl/6 strain. The right common carotid was ligated at P7, P10, P12 or P21 in CD1 mice or at P12 in C57Bl/6 mice. Littermate controls received sham surgery. Seizures were rated for 4h after surgery; brain injury was scored one week later. In a separate group of P12 CD1 mice, electrocorticographic activity was recorded continuously for 4h after carotid ligation or sham surgery. Brain injury and cumulative seizure score varied significantly with age (p <0.001) and strain (p <0.001). In CD1 mice, injury was greatest after ligation on P10 to P12 and seizure score was maximal at P12. Seizure scores were significantly correlated with injury after ligation on P10 or P12. C57Bl/6 mice, like C3Heb/FeJ mice examined previously, were much less vulnerable to seizures and injury than CD1 mice after ligation on P12. This study demonstrates that carotid ligation in the CD1 mouse on P12 causes acute electrographic rhythmic discharges that correlate with behavioral seizures. We also found that the age at which ligation is performed and genetic strain have a strong influence on the severity of injury. [Copyright &y& Elsevier]

Cerebral palsy in humans results from a diverse group of disorders that produce nonprogressive motor impairments in the developing brain. Stroke is an important cause of hemiparetic cerebral palsy in neonates and young children. We recently developed a new immature mouse model of stroke that demonstrates seizures, the severity of which correlates with brain injury. This model has strengths compared with other immature rodent models of ischemic injury, such as relative technical ease and the presence of seizures, as is seen in humans. This model also has relative weaknesses, such as the inability to titrate the severity of the injury with different periods of hypoxia. In addition, more work is needed to delineate the long-term consequences of the insult in this new model. [ABSTRACT FROM AUTHOR]

Neonatal stroke is an important cause of neurologic morbidity and cerebral palsy. Recently, we have determined that in postnatal day 12 CD1 mice unilateral carotid ligation alone results in seizures and brain injury. We have shown that, in this model, seizure scores correlate with brain injury scores. We have applied this model to another strain of mice to assess strain-related differences in vulnerability to seizures and brain injury after unilateral carotid ligation. Under isoflurane anesthesia, unilateral right-sided carotid ligation was performed in postnatal day 12 C3HeB/FeJ mice followed by a 4-hour period of observation in a 35°C incubator. Seizure scores and brain jury scores were assigned and compared to scores in mice receiving sham surgery. Timing of seizure onset and regional distribution of brain injury were compared in the CD1 and C3HeB/FeJ mice. Unilateral carotid ligation in postnatal day 12 C3HeB/FeJ mice resulted in seizure behavior and brain injury in some animals, with similar time to seizure onset and regional injury distribution, but affected a significantly smaller percentage of C3HeB/FeJ pups than that observed in postnatal day 12 CD1 mice, indicating strain-related vulnerability in this model. Copyright © 2005 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Poly(ADP-ribose) polymerase-1 is over-activated in the adult brain in response to ischemia and contributes to neuronal death, but its role in perinatal brain injury remains uncertain. To address this issue, 7-day-old wild-type ( wt ) and PARP-1 gene deficient ( parp+/– and parp–/–) Sv129/CD-1 hybrid mice were subjected to unilateral hypoxia-ischemia and histologic damage was assessed 10 days later by two evaluators. Poly(ADP-ribose) polymerase-1 knockout produced moderate but significant ( p < 0.05) protection in the total group of animals, but analysis by sex revealed that males were strongly protected ( p < 0.05) in contrast to females in which there was no significant effect. Separate experiments demonstrated that PARP-1 was activated over 1–24 h in both females and males after the insult in neonatal wt mice and rats using immnocytochemistry and western blotting for poly(ADP-ribose). Brain levels of NAD+ were also significantly reduced, but the decrease of NAD+ during the early post-hypoxia-ischemia (HI) phase was only seen in males. The results indicate that hypoxia-ischemia activates Poly(ADP-ribose) polymerase-1 in the neonatal brain and that the sex of the animal strongly influences its role in the pathogenesis of brain injury. [ABSTRACT FROM AUTHOR]

Background. Pharmacological openers of mitochondrial ATP-sensitive potassium (mitoKATP) channels have been shown to mimic ischemic preconditioning (IPC) in both the brain and myocardium. We hypothesized that similar endogenous mechanisms exist in the spinal cord and that diazoxide, a potent mitoKATP opener, could reduce neurologic injury after aortic cross-clamping in a model of spinal cord ischemia.Methods. The infra-renal aorta was cross-clamped in 45 male New Zealand white rabbits for 20 minutes. Control animals received no pretreatment. Diazoxide-treated animals were dosed (5 mg/kg) 15 minutes before cross-clamp. A third group underwent 5 minutes of IPC 30 minutes before cross-clamp. Two groups received KATP antagonists, 5-hydroxydecanoic acid (5-HD, 20 mg/kg) or glibenclamide (1.0 mg/kg), before diazoxide administration. Systemic hypotension was induced in a final group with excess isoflurane. Tarlov Scoring was used to assess neurologic function at 24 and 48 hours, after which, the spinal cords were procured for histopathological analysis.Results. Tarlov scoring demonstrated marked improvement in the Diazoxide group compared with control at 24 hours (p < 0.02) and 48 hours (p < 0.009). Moreover, no further neurologic injury occurred in this group at 7 days. IPC-treated animals showed neurologic improvement but were not significantly different from controls. Further, administration of glibenclamide was effective in antagonizing diazoxide’s protective effect.Conclusions. Administration of diazoxide resulted in significant improvement in neurologic outcome in this model. This protective effect improved outcome at both early and late time points. Further, the antagonistic effect of glibenclamide implicates diazoxide’s ATP-dependent potassium channel agonism as the mechanism of protection. Overall, this study suggests that diazoxide may be useful in the prevention of neurologic injury after thoracic aneurysm surgery. [Copyright &y& Elsevier]

Ischemia in the developing brain triggers a series of events that cause damage after a threshold of severity and duration are reached. Release of glutamate, the major excitatory amino acid neurotransmitter in the brain, is one of the primary events in this cascade of injury. Release of glutamate combined with reversal of glutamate uptake pumps in nerve terminals causes glutamate levels to increase from 10-1000x baseline levels in the brains extracellular space during ischemia, traumatic brain injury, and hypoglycemia in animal models and in humans. High levels of extracellular glutamate trigger opening of N-methyl-D-aspartate (NMDA) and non-NMDA glutamate channels, allowing calcium and sodium to flood into neurons. Energy failure in mitochondria facilitates opening of NMDA channels by lowering the membrane potential and reducing their blockade by magnesium that resides Within the channel at normal membrane potentials. Calcium entry into the cytoplasm triggers several events including activation of lipase's, proteases, and nucleuses that can destroy the neuron's cellular machinery. Generation of reactive oxygen free radicals including the messenger molecule nitric oxide can have additional destructive effects. Drugs that block glutamate receptors and the excitotoxin cascade" of down- stream events can protect the brain from injury and show promise for Clinical application in infants and children. [ABSTRACT FROM AUTHOR]

The authors comment on a study published within the issue which used the hypoxia agent Tc-99m-HL9 for the early detection of neuronal injury in a neonatal rat model of hypoxic ischemia. They believe that the greatest advantage of the technique proposed in the study would be the ability to detect salvageable cells that could potentially be rescued by therapeutic interventions. They added that it is also beneficial in patients where the approximate time of a hypoxic-ischemic event is known.

Summary: Neonatal stroke presents with seizures that are usually treated with phenobarbital. We hypothesized that anticonvulsants would attenuate ischemic injury, but that the dose-dependent effects of standard anticonvulsants would impact important age-dependent and injury-dependent consequences. In this study, ischemia induced by unilateral carotid ligation in postnatal day 12 (P12) CD1 mice was immediately followed by an i.p. dose of vehicle, low-dose or high-dose phenobarbital. Severity of acute behavioral seizures was scored. 5-Bromo-2′-deoxyuridine (BrdU) was administered from P18 to P20, behavioral testing performed, and mice perfused at P40. Atrophy quantification and counts of BrdU/NeuN-labeled cells in the dentate gyrus were performed. Blood phenobarbital concentrations were measured. 30mg/kg phenobarbital reduced acute seizures and chronic brain injury, and restored normal weight gain and exploratory behavior. By comparison, 60mg/kg was a less efficacious anticonvulsant, was not neuroprotective, did not restore normal weight gain, and impaired behavioral and cognitive recovery. Hippocampal neurogenesis was not different between treatment groups. These results suggest a protective effect of lower-dose phenobarbital, but a lack of this effect at higher concentrations after stroke in P12 mice. [ABSTRACT FROM AUTHOR]

Ischemic brain injury from stroke is an important cause of disability in infants and children, but current experimental models for the disorder are complex. These preparations require occlusion of small intracerebral vessels or common carotid artery ligation combined with exposure to reduced levels of oxygen. Unilateral carotid artery ligation alone was sufficient to cause brain injury in more than 70% of 12-day-old CD1 mice.Using a blinded behavioral rating scale of seizure activity in mice, a direct, highly significant correlation between the severity of seizures over the 4-hour period after ligation and the severity of histologic brain injury 7 days later (Spearman''s rho = 0.835, P < 0.001) was documented. This study presents the first model of stroke in immature mice produced by unilateral carotid artery ligation alone, and the first to demonstrate a clear correlation between acute ischemia-induced seizures and brain injury. This new model should be useful for examining the pathogenesis of stroke in the immature brain and the potential contribution of seizures to final outcome. [Copyright &y& Elsevier]

Hypoxic-ischemic encephalopathy (HIE) is a leading cause of mortality and morbidity during the perinatal period, and currently no therapeutic drug is available. Minocycline, an antibiotic, has recently been shown to have neuroprotective effects distinct from its antimicrobial effect in several neurological disorders including ischemic brain injury. We examined the effect of minocycline on neonatal hypoxic-ischemic brain injury by using histologic scoring in both mouse and rat models. Mouse (C57Bl/6) and rat (SD) pups were exposed to a unilateral hypoxic-ischemic insult at 8 and 7 days of age, respectively. Minocycline hydrochloride was administered according to protocols that were reported to provide neuroprotection in adult or neonatal rats. Seven days after the insult, we examined brain injury in Nissl stained sections. Although minocycline ameliorated brain injury in the developing rat, it increased injury in the developing mouse. This detrimental effect in the mouse was consistent across different regions (cortex, striatum, and thalamus), with both single and multiple injection protocols and with both moderate and high-dose treatment (P < 0.05). The mechanism of the contrasting effects in mouse and rat is not clear and remains to be elucidated. Minocycline has been used as an antibiotic in the clinical setting for decades; therefore, it may be considered for use in infants with hypoxic-ischemic brain damage, based on prior reports of neuroprotection in the rat. However, it is important to examine this drug carefully before clinical use in human infants, taking our data in the mouse model into consideration. [Copyright &y& Elsevier]

A consecutive case series of 55 children (0-17 years old) with arterial ischemic stroke is reported. Twenty of these children were critically ill at the time their stroke occurred. Mortality among these 20 children was 40%, compared with 3% in non-critically ill children with arterial ischemic stroke; overall mortality in this case series was 16%. Mortality resulted primarily from the underlying illness. Prognosis after stroke is markedly worsened in children with premorbid critical illness. [Copyright &y& Elsevier]