Diffusion tensor imaging is highly sensitive to the microstructural integrity of the brain and has uncovered significant abnormalities following traumatic brain injury not appreciated through other methods. It is hoped that this increased sensitivity will aid in the detection and prognostication in patients with traumatic injury. However, the pathological substrates of such changes are poorly understood. Specifically, decreases in fractional anisotropy derived from diffusion tensor imaging are consistent with axonal injury, myelin injury or both in white matter fibres. In contrast, in both humans and animal models, increases in fractional anisotropy have been suggested to reflect axonal regeneration and plasticity, but the direct histological evidence for such changes remains tenuous. We developed a method to quantify the anisotropy of stained histological sections using Fourier analysis, and applied the method to a rat controlled cortical impact model to identify the specific pathological features that give rise to the diffusion tensor imaging changes in subacute to chronic traumatic brain injury. A multiple linear regression was performed to relate the histological measurements to the measured diffusion tensor changes. The results show that anisotropy was significantly increased (P < 0.001) in the perilesioned cortex following injury. Cortical anisotropy was independently associated (standardized β = 0.62, P = 0.04) with the coherent organization of reactive astrocytes (i.e. gliosis) and was not attributed to axons. By comparison, a decrease in white matter anisotropy (P < 0.001) was significantly related to demyelination (β = 0.75, P = 0.0015) and to a lesser extent, axonal degeneration (β = -0.48, P = 0.043). Gliosis within the lesioned cortex also influenced diffusion tensor tractography, highlighting the fact that spurious tracts in the injured brain may not necessarily reflect continuous axons and may instead depict glial scarring. The current study demonstrates a novel method to relate pathology to diffusion tensor imaging findings, elucidates the underlying mechanisms of anisotropy changes following traumatic brain injury and significantly impacts the clinical interpretation of diffusion tensor imaging findings in the injured brain.

Stroke is the leading cause of disability in the United States and affects 15 million people worldwide. Studies performed in various parts of the world have found differences between sexes in stroke incidence, prevalence, mortality, and outcomes. Although men are at higher risk of stroke for most age groups below age 85 years, after this age the incidence reverses dramatically, with women being much more at risk. Furthermore, recent studies suggest that women have worse recovery than men post-stroke. Many aspects of recovery may influence this outcome, including sex-specific comorbidities, aggressiveness of acute treat-ment, prevention therapies, and varying degrees of social support and rates of depression. It is important to further define and investigate sex differences in stroke incidence, care, treatment, and outcomes to improve functional recovery in women.

Increasing evidence suggests that sex differences exist in the etiology, presentation, treatment, and outcome from stroke. The reasons for these sex disparities are becoming increasingly explored, but large gaps still exist in our knowledge. Experimental studies over the past several years have demonstrated intrinsic sex differences both in vivo and in animal models which may have relevance to our understanding of stroke in clinical populations. A greater understanding of the differences and similarities between males and females with respect to the risk factors, pathophysiology, and response to stroke will facilitate the design of future clinical trials and enhance the development of treatment strategies to improve stroke care in both sexes. This article reviews the current literature on sex differences in stroke with an emphasis on the clinical data, incorporating an analysis of bench research as it pertains to the bedside.

BackgroundAfter central nervous system injury, inflammatory macrophages (M1) predominate over anti-inflammatory macrophages (M2). The temporal profile of M1/M2 phenotypes in macrophages and microglia after traumatic brain injury (TBI) in rats is unknown. We subjected female rats to severe controlled cortical impact (CCI) and examined the postinjury M1/M2 time course in their brains.MethodsThe motor cortex (2.5 mm left laterally and 1.0 mm anteriorly from the bregma) of anesthetized female Wistar rats (ages 8 to 10 weeks; N = 72) underwent histologically moderate to severe CCI with a 5-mm impactor tip. Separate cohorts of rats had their brains dissociated into cells for flow cytometry, perfusion-fixed for immunohistochemistry (IHC) and ex vivo magnetic resonance imaging or flash-frozen for RNA and protein analysis. For each analytical method used, separate postinjury times were included for 24 hours; 3 or 5 days; or 1, 2, 4 or 8 weeks.ResultsBy IHC, we found that the macrophagic and microglial responses peaked at 5 to 7 days post-TBI with characteristics of mixed populations of M1 and M2 phenotypes. Upon flow cytometry examination of immunological cells isolated from brain tissue, we observed that peak M2-associated staining occurred at 5 days post-TBI. Chemokine analysis by multiplex assay showed statistically significant increases in macrophage inflammatory protein 1α and keratinocyte chemoattractant/growth-related oncogene on the ipsilateral side within the first 24 hours after injury relative to controls and to the contralateral side. Quantitative RT-PCR analysis demonstrated expression of both M1- and M2-associated markers, which peaked at 5 days post-TBI.ConclusionsThe responses of macrophagic and microglial cells to histologically severe CCI in the female rat are maximal between days 3 and 7 postinjury. The response to injury is a mixture of M1 and M2 phenotypes.

Objective Metrics of diffusion tensor imaging (DTI) and magnetization transfer imaging (MTI) can detect diffuse axonal injury in traumatic brain injury (TBI). The relationship between the changes of these imaging measures and the underlying pathologies is still relatively unknown. This study investigated the radiological-pathological correlation between these imaging techniques and immunohistochemistry using a closed head rat model of TBI. Methods TBI was performed on female rats followed longitudinally by MRI out to 30 days post-injury, with a subset of animals selected for histopathological analyses. A MRI-based finite element analysis was generated to characterize the pattern of the mechanical insult and estimate the extent of brain injury to direct the pathological correlation with imaging findings. Results DTI axial diffusivity and fractional anisotropy (FA) were sensitive to axonal integrity, while radial diffusivity showed significant correlation to the myelin compactness. FA was correlated to astrogliosis in the gray matter while mean diffusivity was correlated to increased cellularity. Secondary inflammatory responses also partly affected the changes of these DTI metrics. The magnetization transfer ratio (MTR) at 3.5 ppm demonstrated a strong correlation with both axon and myelin integrity. Decrease in MTR at 20 ppm correlated with the extent of astrogliosis in both gray and white matter. Interpretation While conventional T2-weighted MRI did not detect abnormalities following TBI, DTI and MTI afforded complementary insight into the underlying pathologies reflecting varying injury states over time, thus may substitute for histology to reveal DAI pathologies in vivo. This correlation of MRI and histology furthers understanding of the microscopic pathology underlying DTI and MTI changes in TBI.

Traumatic microbleeds are a common neuroimaging finding in traumatic brain injury, yet their clinical significance remains unclear. Griffin et al. report that traumatic microbleeds predict disability, and use MRI-guided histopathology to elucidate the underlying pathophysiology. They conclude that traumatic microbleeds may be a form of traumatic vascular injury.

White adipose tissue plays an integral role in energy metabolism and is governed by endocrine, autocrine, and neural signals. Neural control of adipose metabolism is mediated by sympathetic neurons that innervate the tissue. To investigate the effects of this innervation, an ex vivo system was developed in which 3T3-L1 adipocytes are cocultured with sympathetic neurons isolated from the superior cervical ganglia of newborn rats. In coculture, both adipocytes and neurons exhibit appropriate morphology, express cell-type-specific markers, and modulate key metabolic processes in one another. Lipolysis (stimulated by ␤-adrenergic agents) and leptin secretion by adipocytes are down-regulated by neurons in coculture, effects apparently mediated by neuropeptide Y (NPY). Secretion of NPY by neurons is up-regulated dramatically by the presence of adipocytes in coculture and appears to be mediated by an adipocyte-derived soluble factor. Insulin, an antilipolytic agent, down-regulates NPY secretion. Our findings suggest that an adipocyte-derived factor(s) up-regulates the secretion of NPY by sympathetic neurons, which, in turn, attenuates lipolytic energy mobilization by adipocytes.A dipose tissue functions not only as a storage site for triglyceride energy reserves, but also as the source of numerous endocrine (1-3) and paracrine factors (4). The excessive accumulation of adipose tissue as occurs in obesity (5) or too little of this tissue as occurs in lipodystrophy (6) leads to major human health problems that are increasing in frequency. For these reasons, there has been increased interest in understanding how endocrine, paracrine, and neural factors control the metabolism of white adipose tissue (WAT). Least understood are the neural factors that regulate the metabolism of WAT.The primary role of adipose tissue is to store energy as triglyceride when nutritional fuel is abundant and to mobilize that energy reserve when stimulated appropriately. In WAT, stored triglyceride is mobilized by the hormone-sensitive lipase to release and export free fatty acids and glycerol to provide physiological fuel for other tissues. In contrast, the lipid in brown adipose tissue (BAT) is used predominantly for heat production. Lipolysis is triggered by a variety of hormonal and neural effectors that increase the cAMP level in adipocytes, including catecholamines (epinephrine and norepinephrine) that interact with ␤-adrenergic receptors (7). While circulating catecholamine (epinephrine) regulates lipolysis in adipose tissue, particularly in the fasted state, catecholamine (notably norepinephrine) also is released from sympathetic neurons that innervate WAT (8).Sympathetic fibers of the autonomic nervous system innervate both white and brown adipose tissue (9). Histological evidence gained from studies with rodents and pigs has shown that adipose tissue is innervated by the sympathetic neurons and is highly vascularized, with every adipocyte in close proximity to a capillary (10). Early studies (11, 12) suggested that the major innervatio...

Acute traumatic brain injury (TBI) is associated with long-term cognitive and behavioral dysfunction. In vivo studies have shown histone deacetylase inhibitors (HDACis) to be neuroprotective following TBI in rodent models. HDACis are intriguing candidates because they are capable of provoking widespread genetic changes and modulation of protein function. By using known HDACis and a unique small-molecule pan-HDACi (LB-205), we investigated the effects and mechanisms associated with HDACi-induced neuroprotection following CNS injury in an astrocyte scratch assay in vitro and a rat TBI model in vivo. We demonstrate the preservation of sufficient expression of nerve growth factor (NGF) and activation of the neurotrophic tyrosine kinase receptor type 1 (TrkA) pathway following HDACi treatment to be crucial in stimulating the survival of CNS cells after TBI. HDACi treatment up-regulated the expression of NGF, phospho-TrkA, phospho-protein kinase B (p-AKT), NF-κB, and B-cell lymphoma 2 (Bcl-2) cell survival factors while down-regulating the expression of p75 neurotrophin receptor (NTR), phospho-JNK, and Bcl-2-associated X protein apoptosis factors. HDACi treatment also increased the expression of the stem cell biomarker nestin, and decreased the expression of reactive astrocyte biomarker GFAP within damaged tissue following TBI. These findings provide further insight into the mechanisms by which HDACi treatment after TBI is neuroprotective and support the continued study of HDACis following acute TBI.A cute traumatic brain injury (TBI) is a leading cause of disability and death, and results in reduced quality of life for surviving patients and prolonged economic effects on society (1). Primary injury occurs at the moment of trauma and is the direct result of shearing, tearing, and stretching of the brain parenchyma affecting neural tissue and blood vessels, and causing immediate contusion, hemorrhage, diffuse axonal injury, and ischemia (2, 3). TBI brings about cognitive and behavioral dysfunction through a complex sequence of secondary pathologic changes following injury. The mechanisms evolve over time and include excessive neurotransmitter release, mitochondrial dysfunction, increased bloodbrain barrier (BBB) permeability, cerebral edema, inflammation, and seizures, causing cell death and clinical morbidity (4).Clinical trials designed to test therapeutic agents aimed at improving residual cerebral function after TBI have not been successful because of the multifactorial nature of the disorder (2, 5-7). No pharmacological agent is currently approved for the treatment of acute TBI. The present standard of care consists of maintaining physiologic function by preserving adequate cerebral perfusion and normal intracranial pressure. Histone deacetylase inhibitors (HDACis) are potential candidates for the treatment of TBI. They are capable of inducing widespread alterations in cellular function and protein expression through posttranslational modification of histones, transcriptional factors, and heat shock chaperones (8-...