Blood-Brain Barrier Dysfunction and Exposure to Head Impacts in University Football Players.

OBJECTIVE: To investigate the link between dysfunction of the blood-brain barrier (BBB) and exposure to head impacts in concussed football athletes. DESIGN: This was a prospective, observational pilot study. SETTING: Canadian university football. PARTICIPANTS: The study population consisted of 60 university football players, aged 18 to 25. Athletes who sustained a clinically diagnosed concussion over the course of a single football season were invited to undergo an assessment of BBB leakage. INDEPENDENT VARIABLES: Head impacts detected using impact-sensing helmets were the measured variables. MAIN OUTCOME MEASURES: Clinical diagnosis of concussion and BBB leakage assessed using dynamic contrast-enhanced MRI (DCE-MRI) within 1 week of concussion were the outcome measures. RESULTS: Eight athletes were diagnosed with a concussion throughout the season. These athletes sustained a significantly higher number of head impacts than nonconcussed athletes. Athletes playing in the defensive back position were significantly more likely to sustain a concussion than remain concussion free. Five of the concussed athletes underwent an assessment of BBB leakage. Logistic regression analysis indicated that region-specific BBB leakage in these 5 athletes was best predicted by impacts sustained in all games and practices leading up to the concussion-as opposed to the last preconcussion impact or the impacts sustained during the game when concussion occurred. CONCLUSIONS: These preliminary findings raise the potential for the hypothesis that repeated exposure to head impacts may contribute to the development of BBB pathology. Further research is needed to validate this hypothesis and to test whether BBB pathology plays a role in the sequela of repeated head trauma.

Concussion susceptibility is mediated by spreading depolarization-induced neurovascular dysfunction.

The mechanisms underlying the complications of mild traumatic brain injury, including post-concussion syndrome, post-impact catastrophic death, and delayed neurodegeneration remain poorly understood. This limited pathophysiological understanding has hindered the development of diagnostic and prognostic biomarkers and has prevented the advancement of treatments for the sequelae of mild traumatic brain injury. We aimed to characterize the early electrophysiological and neurovascular alterations following repetitive mild traumatic brain injury and sought to identify new targets for the diagnosis and treatment of individuals at risk of severe post-impact complications. We combined behavioural, electrophysiological, molecular, and neuroimaging techniques in a rodent model of repetitive mild traumatic brain injury. In humans, we used dynamic contrast-enhanced MRI to quantify blood-brain barrier dysfunction after exposure to sport-related concussive mild traumatic brain injury. Rats could clearly be classified based on their susceptibility to neurological complications, including life-threatening outcomes, following repetitive injury. Susceptible animals showed greater neurological complications and had higher levels of blood-brain barrier dysfunction, transforming growth factor ß (TGFß) signalling, and neuroinflammation compared to resilient animals. Cortical spreading depolarizations were the most common electrophysiological events immediately following mild traumatic brain injury and were associated with longer recovery from impact. Triggering cortical spreading depolarizations in mild traumatic brain injured rats (but not in controls) induced blood-brain barrier dysfunction. Treatment with a selective TGFß receptor inhibitor prevented blood-brain barrier opening and reduced injury complications. Consistent with the rodent model, blood-brain barrier dysfunction was found in a subset of human athletes following concussive mild traumatic brain injury. We provide evidence that cortical spreading depolarization, blood-brain barrier dysfunction, and pro-inflammatory TGFß signalling are associated with severe, potentially life-threatening outcomes following repetitive mild traumatic brain injury. Diagnostic-coupled targeting of TGFß signalling may be a novel strategy in treating mild traumatic brain injury.

Microbiome differential abundance methods produce different results across 38 datasets.

Identifying differentially abundant microbes is a common goal of microbiome studies. Multiple methods are used interchangeably for this purpose in the literature. Yet, there are few large-scale studies systematically exploring the appropriateness of using these tools interchangeably, and the scale and significance of the differences between them. Here, we compare the performance of 14 differential abundance testing methods on 38 16S rRNA gene datasets with two sample groups. We test for differences in amplicon sequence variants and operational taxonomic units (ASVs) between these groups. Our findings confirm that these tools identified drastically different numbers and sets of significant ASVs, and that results depend on data pre-processing. For many tools the number of features identified correlate with aspects of the data, such as sample size, sequencing depth, and effect size of community differences. ALDEx2 and ANCOM-II produce the most consistent results across studies and agree best with the intersect of results from different approaches. Nevertheless, we recommend that researchers should use a consensus approach based on multiple differential abundance methods to help ensure robust biological interpretations.

Bacterial Taxa and Functions Are Predictive of Sustained Remission Following Exclusive Enteral Nutrition in Pediatric Crohn's Disease.

BACKGROUND: The gut microbiome is extensively involved in induction of remission in pediatric Crohn's disease (CD) patients by exclusive enteral nutrition (EEN). In this follow-up study of pediatric CD patients undergoing treatment with EEN, we employ machine learning models trained on baseline gut microbiome data to distinguish patients who achieved and sustained remission (SR) from those who did not achieve remission nor relapse (non-SR) by 24 weeks. METHODS: A total of 139 fecal samples were obtained from 22 patients (8-15 years of age) for up to 96 weeks. Gut microbiome taxonomy was assessed by 16S rRNA gene sequencing, and functional capacity was assessed by metagenomic sequencing. We used standard metrics of diversity and taxonomy to quantify differences between SR and non-SR patients and to associate gut microbial shifts with fecal calprotectin (FCP), and disease severity as defined by weighted Pediatric Crohn's Disease Activity Index. We used microbial data sets in addition to clinical metadata in random forests (RFs) models to classify treatment response and predict FCP levels. RESULTS: Microbial diversity did not change after EEN, but species richness was lower in low-FCP samples (<250 µg/g). An RF model using microbial abundances, species richness, and Paris disease classification was the best at classifying treatment response (area under the curve [AUC] = 0.9). KEGG Pathways also significantly classified treatment response with the addition of the same clinical data (AUC = 0.8). Top features of the RF model are consistent with previously identified IBD taxa, such as Ruminococcaceae and Ruminococcus gnavus. CONCLUSIONS: Our machine learning approach is able to distinguish SR and non-SR samples using baseline microbiome and clinical data.

Multi-omics differentially classify disease state and treatment outcome in pediatric Crohn's disease.

BACKGROUND: Crohn's disease (CD) has an unclear etiology, but there is growing evidence of a direct link with a dysbiotic microbiome. Many gut microbes have previously been associated with CD, but these have mainly been confounded with patients' ongoing treatments. Additionally, most analyses of CD patients' microbiomes have focused on microbes in stool samples, which yield different insights than profiling biopsy samples. RESULTS: We sequenced the 16S rRNA gene (16S) and carried out shotgun metagenomics (MGS) from the intestinal biopsies of 20 treatment-naïve CD and 20 control pediatric patients. We identified the abundances of microbial taxa and inferred functional categories within each dataset. We also identified known human genetic variants from the MGS data. We then used a machine learning approach to determine the classification accuracy when these datasets, collapsed to different hierarchical groupings, were used independently to classify patients by disease state and by CD patients' response to treatment. We found that 16S-identified microbes could classify patients with higher accuracy in both cases. Based on follow-ups with these patients, we identified which microbes and functions were best for predicting disease state and response to treatment, including several previously identified markers. By combining the top features from all significant models into a single model, we could compare the relative importance of these predictive features. We found that 16S-identified microbes are the best predictors of CD state whereas MGS-identified markers perform best for classifying treatment response. CONCLUSIONS: We demonstrate for the first time that useful predictors of CD treatment response can be produced from shotgun MGS sequencing of biopsy samples despite the complications related to large proportions of host DNA. The top predictive features that we identified in this study could be useful for building an improved classifier for CD and treatment response based on sufferers' microbiome in the future. The BISCUIT project is funded by a Clinical Academic Fellowship from the Chief Scientist Office (Scotland)-CAF/08/01.

The effects of aging on neuron number in the rat superior olivary complex.

The superior olivary complex (SOC) is an auditory center in the brainstem involved in the processing of binaural information. The ability to localize sounds is dependent on the efficient processing of binaural signals by the SOC and other auditory centers. It has been demonstrated that sound localization ability is impaired in aged rats, and morphological changes in the SOC may contribute to this deficit. In the present study, neuron counts were done on three key nuclei of the SOC: the medial nucleus of the trapezoid body (MNTB), the lateral superior olivary nucleus (LSO), and the medial superior olivary nucleus (MSO) in groups of Fischer 344 rats aged 3, 12, 24, and 30 months. Neuron number remains stable between 3 and 30 months of age in the LSO and MSO, however, in the MNTB, neuron number is significantly reduced at 24 and 30 months of age. Neuron loss in the MNTB of 24-month-old Fischer 344 rats is not as heavy as that reported earlier in 24-month-old Sprague-Dawley rats (8% loss versus 34% loss), indicating a strain difference with regard to aging in the SOC.

Aging in the rat medial nucleus of the trapezoid body. III. Alterations in capillaries.

The medial nucleus of the trapezoid body (MTB), a large cell group in the rat brainstem auditory pathway, undergoes significant cell loss and loss of synapses with advancing age [5,6]. The purpose of the present study was to examine the microvasculature of the MTB in rats of the following ages: 3 months (MO), 6 MO, 24 MO, 27 MO, and 33 MO. In rats aged 24 to 33 MO, the following ultrastructural changes were observed in MTB capillaries: (1) large cavitations or spaces within capillary basal laminae, and (2) membranous debris, indicative of cellular degeneration within leaflets of capillary basal lamina. The volume density ratio (VDR) of capillaries decreases significantly between 6 and 33 MO of age.

Aging in the rat medial nucleus of the trapezoid body. I. Light microscopy.

The medial nucleus of the trapezoid body (MTB), a cell group in the superior olivary complex, was examined in an age-graded series of rats for neuron loss, changes in the giant synaptic endings (chalices of Held) on MTB neurons, and accumulation of age pigment. Neuron counts were done on protargol-stained paraffin sections of MTBs from a series of 17 rats aged 2-3, 6, 18, and 24 months. Between 2-3 and 24 months a 34% decrease in the mean number of MTB neurons was observed. Significant loss (p less than 0.05) was first evident in the early portion of the life span, between 2-3 and 6 months. In thionin-stained sections, there was no change with aging in the proportions of three MTB neuron types: principal cells (approximately 82%), elongate cells (approximately 15%), and stellate cells (approximately 3%). In young adult rats, 25-26% of all MTB neurons were associated with identifiable chalices of Held in protargol stained sections. This ratio did not vary significantly with aging. Age pigment accumulation in the MTB was examined in 2 micrometers Araldite sections stained with toluidine blue. Age pigment deposits were larger and more numerous in the MTBs of old animals, but not as extensive as has been described previously in many other parts of the nervous system. This study is the first to report neuron loss in an animal brainstem nucleus.

Aging in the rat medial nucleus of the trapezoid body. II. Electron microscopy.

The medial nucleus of the trapezoid body (MTB), the largest cell group of the rat superior olivary complex, was studied ultrastructurally in five groups of rats ranging in age from 2 to 33 months. Relatively little aging pigment (lipofuscin) accumulates intraneuronally, even in rats aged 33 months. The heaviest accumulation of age pigment is in glial and perivascular cells of the MTB. In rats aged 24-33 months, there is evidence of axonal and dendritic degeneration in the MTB neuropil. Aged rats also show evidence of degeneration of nerve endings. This includes the loss of calycine axosomatic terminals synapsing with MTB principal cells. All of the ultrastructural changes observed in the MTB gradually increase in frequency with advancing age.

Age-related loss of synaptic terminals in the rat medial nucleus of the trapezoid body.

The effect of aging on axosomatic synaptic terminals in the rat medial nucleus of the trapezoid body was studied using quantitative electron microscopy. In young adult rats (3 months of age), the mean percentage of the surface area of principal cells covered by synaptic terminals is 61.7% (S.E.M. = 4.1) while in aged animals (27-33 months of age) the per cent coverage is 43.7% (S.E.M. = 3.3). Likewise, between 3 and 27-33 months of age, the average number of synaptic terminals present along a 100 micron length of principal cell surface decreases significantly (P less than 0.001) from 28.3 (S.E.M. = 1.3) to 18.9 (S.E.M. = 1.3). Only terminals derived from calyces of Held are lost in the aged animals, displaying a 37% reduction between 3 and 27-33 months of age. The length of apposition by synaptic terminals in the medial nucleus of the trapezoid body does not change significantly with aging. We conclude that because of a significant loss of calycine synaptic endings, the structure of calyces of Held becomes less complex with advancing age in rats. This would presumably result in an age-related partial deafferentation of principal cells, causing significant alterations in the processing of auditory information in the medial nucleus of the trapezoid body.

Factors influencing food choices of adolescents: findings from focus-group discussions with adolescents.

OBJECTIVE: To assess adolescents' perceptions about factors influencing their food choices and eating behaviors. DESIGN: Data were collected in focus-group discussions. SUBJECTS/SETTING: The study population included 141 adolescents in 7th and 10th grade from 2 urban schools in St Paul, Minn, who participated in 21 focus groups. ANALYSIS: Data were analyzed using qualitative research methodology, specifically, the constant comparative method. RESULTS: Factors perceived as influencing food choices included hunger and food cravings, appeal of food, time considerations of adolescents and parents, convenience of food, food availability, parental influence on eating behaviors (including the culture or religion of the family), benefits of foods (including health), situation-specific factors, mood, body image, habit, cost, media, and vegetarian beliefs. Major barriers to eating more fruits, vegetables, and dairy products and eating fewer high-fat foods included a lack of sense of urgency about personal health in relation to other concerns, and taste preferences for other foods. Suggestions for helping adolescents eat a more healthful diet include making healthful food taste and look better, limiting the availability of unhealthful options, making healthful food more available and convenient, teaching children good eating habits at an early age, and changing social norms to make it "cool" to eat healthfully. APPLICATIONS/CONCLUSIONS: The findings suggest that if programs to improve adolescent nutrition are to be effective, they need to address a broad range of factors, in particular environmental factors (e.g., the increased availability and promotion of appealing, convenient foods within homes schools, and restaurants).

IBM correcting selectric typewriter: an analysis of the use of the correctable film ribbon in altering typewritten documents.

From the point of view of a document examiner, the advent of the Correcting Selectric typewriter has not changed any of the procedures ordinarily used in the examination of altered typewritten material. The same precautions should be observed in conducting examinations of such material as would be observed in making examinations for any other type of possible alteration on negotiable instruments. The security question raised with the Correcting Selectric is no different than the questions raised when an eraser was put on a pencil or a chemical bleach was found to remove ink on paper. The Correcting Selectric might even be compared to the introduction of ball-point pens on the market in the late 1940s. At that time, financial publications were suggesting that ball-point pens should not be used on any negotiable or valuable document as the ink was considered to be too readily transferable, and it was thought that they left little in the way of individual writing characteristics. However, it was not long before financial institutions were supplying such pens on their counters. Because of the complexities involved in using the Correcting Selectric to alter a document, it is felt that such problems will prove to be less prevalent than those involving other means of alterations.