Association of mild traumatic brain injury, post-traumatic stress disorder, and other comorbidities on photosensitivity.

SIGNIFICANCE: Photosensitivity is common after mild traumatic brain injury. However, this study demonstrates that photosensitivity is also impacted by common comorbidities that often occur with mild traumatic brain injury. Understanding how physical and psychological traumas impact photosensitivity can help improve provider care to trauma survivors and guide novel therapeutic interventions. PURPOSE: This study aimed to characterize the association between mild traumatic brain injury and common comorbidities on photosensitivity in post-9/11 veterans. METHODS: Existing data from the Translational Research Center for TBI and Stress Disorders cohort study were analyzed including traumatic brain injury history and post-traumatic stress disorder clinical diagnostic interviews; sleep quality, anxiety, and depression symptoms self-report questionnaires; and photosensitivity severity self-report from the Neurobehavioral Symptom Inventory. Analysis of covariance and multiple ordinal regression models were used to assess associations between mild traumatic brain injury and common comorbidities with photosensitivity severity. RESULTS: Six hundred forty-one post-9/11 veterans were included in this study. An initial analysis showed that both mild traumatic brain injury and current post-traumatic stress disorder diagnosis were independently associated with higher photosensitivity ratings compared with veterans without either condition, with no interaction observed between these two conditions. Results of the ordinal regression models demonstrated positive associations between degree of photosensitivity and the number of mild traumatic brain injuries during military service and current post-traumatic stress disorder symptom severity, particularly hyperarousal symptoms, even when controlling for other factors. In addition, the degree of sleep disturbances and current anxiety symptoms were both positively associated with photosensitivity ratings, whereas depression symptoms, age, and sex were not. CONCLUSIONS: Repetitive mild traumatic brain injury, post-traumatic stress disorder, anxiety, and sleep disturbances were all found to significantly impact photosensitivity severity and are therefore important clinical factors that eye care providers should consider when managing veterans with a history of deployment-related trauma reporting photosensitivity symptoms.

Case Report: Visual Snow Syndrome after Repetitive Mild Traumatic Brain Injury.

SIGNIFICANCE: Visual snow syndrome is a recently recognized condition with its own diagnostic criteria, evolving pathophysiologic research, and potential treatment options. PURPOSE: This report documents a rare but likely underdiagnosed condition called visual snow syndrome. A review of the current literature on pathophysiology and treatments is discussed. CASE REPORT: A 40-year-old Whiteman started experiencing symptoms of constant pulsating pixels throughout his entire visual field approximately 3 weeks after a series of mild concussions. In addition, he experienced a persistence of images and photosensitivity. The patient had normal eye examination results, visual fields, and retinal imaging result. Brain MRI, magnetic resonance angiography, electroencephalography, and cerebrospinal fluid analysis were unremarkable. A positron emission tomography scan demonstrated hypometabolism in the posterior parietal lobes and left posterior cingulate gyrus. Pharmacological treatment with antiepileptic and migraine medications was unsuccessful. Tinted lenses were essentially ineffective with a 10% reduction in symptoms reported with the use of a custom blue-tinted lens.Vision rehabilitation aids with optical character recognition were used for prolonged reading needs. CONCLUSIONS: Although rare, visual snow syndrome should be considered in all patients reporting continuous pixelations in their vision for more than 3 months, especially when accompanied by at least two of the following: photosensitivity, palinopsia, enhanced entopic phenomena, or nyctalopia. The pathophysiology is still unclear at this point, with evidence suggesting a link to the secondary visual cortex, specifically the lingual gyrus. More studies are needed to determine the exact cause, especially studies that separate visual snow syndrome patients with and without comorbid migraine. Because the pathophysiology is unclear, the treatment course is also unclear. Anecdotal evidence may suggest that tinted lenses may be of some value.

Blast mild traumatic brain injury is associated with increased myopia and chronic convergence insufficiency.

While chronic visual symptom complaints are common among Veterans with a history of mild traumatic brain injury (mTBI), research is still ongoing to characterize the pattern of visual deficits that is most strongly associated with mTBI and specifically, the impact of blast-related mTBI on visual functioning. One area that has not been well explored is the potential impact of blast mTBI on refractive error. While myopic shifts have been documented following head injuries in civilian populations, posttraumatic myopic shifts have not been explored in participants with military mTBI. This study investigated the impact of blast mTBIs on a range of visual function measures including distance acuity and refractive error, in a well-characterized cohort of thirty-one Post-9/11 veterans for whom detailed clinical interviews regarding military and TBI history were available. Seventeen participants had a history of blast-related mTBI (blast mTBI + group) while 14 did not (blast mTBI- group). Results show an increased frequency of convergence insufficiency and myopia in the blast mTBI + group relative to the blast mTBI- group. Linear regression analyses further show that deficits in distance acuity and refractive error are associated with the number of blast mTBIs during military service but not the number of non-blast mTBIs or the number of lifetime non-blast TBIs and cannot be accounted for by PTSD. These results are consistent with long-lasting damage following blast mTBI to subcortical visual structures that support both vergence movements and the accommodative functions needed to see clearly objects at varying distances.

External validation of a convolutional neural network artificial intelligence tool to predict malignancy in pulmonary nodules.

BACKGROUND: Estimation of the risk of malignancy in pulmonary nodules detected by CT is central in clinical management. The use of artificial intelligence (AI) offers an opportunity to improve risk prediction. Here we compare the performance of an AI algorithm, the lung cancer prediction convolutional neural network (LCP-CNN), with that of the Brock University model, recommended in UK guidelines. METHODS: A dataset of incidentally detected pulmonary nodules measuring 5-15 mm was collected retrospectively from three UK hospitals for use in a validation study. Ground truth diagnosis for each nodule was based on histology (required for any cancer), resolution, stability or (for pulmonary lymph nodes only) expert opinion. There were 1397 nodules in 1187 patients, of which 234 nodules in 229 (19.3%) patients were cancer. Model discrimination and performance statistics at predefined score thresholds were compared between the Brock model and the LCP-CNN. RESULTS: The area under the curve for LCP-CNN was 89.6% (95% CI 87.6 to 91.5), compared with 86.8% (95% CI 84.3 to 89.1) for the Brock model (p≤0.005). Using the LCP-CNN, we found that 24.5% of nodules scored below the lowest cancer nodule score, compared with 10.9% using the Brock score. Using the predefined thresholds, we found that the LCP-CNN gave one false negative (0.4% of cancers), whereas the Brock model gave six (2.5%), while specificity statistics were similar between the two models. CONCLUSION: The LCP-CNN score has better discrimination and allows a larger proportion of benign nodules to be identified without missing cancers than the Brock model. This has the potential to substantially reduce the proportion of surveillance CT scans required and thus save significant resources.

Genetic associations and phenotypic heterogeneity in the craniosynostotic rabbit.

Craniosynostosis (CS) is a disorder that involves the premature ossification of one or more cranial sutures. Our research team has described a naturally occurring rabbit model of CS with a variable phenotype and unknown etiology. Restriction-site associated DNA (RAD) sequencing is a genomic sampling method for identifying genetic variants in species with little or no existing sequence data. RAD sequencing data was analyzed using a mixed linear model to identify single nucleotide polymorphisms (SNPs) associated with disease occurrence and onset in the rabbit model of CS. SNPs achieving a genome-wide significance of p ≤ 5 x 10-8 were identified on chromosome 2 in association with disease occurrence and on chromosomes 14 and 19 in association with disease onset. Genotyping identified a coding variant in fibroblast growth factor binding protein 1 (FGFBP-1) on chromosome 2 and a non-coding variant upstream of integrin alpha 3 (ITGA3) on chromosome 19 that associated with disease occurrence and onset, respectively. Retrospective analysis of patient data revealed a significant inverse correlation between FGFBP-1 and ITGA3 transcript levels in patients with coronal CS. FGFBP-1 and ITGA3 are genes with roles in early development that warrant functional study to further understand suture biology.

Single suture craniosynostosis: Identification of rare variants in genes associated with syndromic forms.

We report RNA-Sequencing results on a cohort of patients with single suture craniosynostosis and demonstrate significant enrichment of heterozygous, rare, and damaging variants among key craniosynostosis-related genes. Genetic burden analysis identified a significant increase in damaging variants in ATR, EFNA4, ERF, MEGF8, SCARF2, and TGFBR2. Of 391 participants, 15% were found to have damaging and potentially causal variants in 29 genes. We observed transmission in 96% of the affected individuals, and thus penetrance, epigenetics, and oligogenic factors need to be considered when recommending genetic testing in patients with nonsyndromic craniosynostosis.

Development and validation of clinical prediction models to risk stratify patients presenting with small pulmonary nodules: a research protocol.

INTRODUCTION: Lung cancer is a common cancer, with over 1.3 million cases worldwide each year. Early diagnosis using computed tomography (CT) screening has been shown to reduce mortality but also detect non-malignant nodules that require follow-up scanning or alternative methods of investigation. Practical and accurate tools that can predict the probability that a lung nodule is benign or malignant will help reduce costs and the risk of morbidity and mortality associated with lung cancer. METHODS: Retrospectively collected data from 1500 patients with pulmonary nodule(s) of up to 15 mm detected on routinely performed CT chest scans aged 18 years old or older from three academic centres in the UK will be used to to develop risk stratification models. Radiological, clinical and patient characteristics will be combined in multivariable logistic regression models to predict nodule malignancy. Data from over 1000 participants recruited in a prospective phase of the study will be used to evaluate model performance. Discrimination, calibration and clinical utility measures will be presented.

Intracranial pressure changes during mouse development.

During early stages of postnatal development, pressure from the growing brain as well as cerebrospinal fluid, i.e. intracranial pressure (ICP), load the calvarial bones. It is likely that such loading contributes to the peripheral bone formation at the sutural edges of calvarial bones, especially shortly after birth when the brain is growing rapidly. The aim of this study was to quantify ICP during mouse development. A custom pressure monitoring system was developed and calibrated. It was then used to measure ICP in a total of seventy three wild type mice at postnatal (P) day 3, 10, 20, 31 and 70. Retrospectively, the sample in each age group with the closest ICP to the average value was scanned using micro-computed tomography to estimate cranial growth. ICP increased from 1.33±0.87mmHg at P3 to 1.92±0.78mmHg at P10 and 3.60±1.08mmHg at P20. In older animals, ICP plateaued at about 4mmHg. There were statistically significant differences between the ICP at the P3 vs. P20, and P10 vs. P20. In the samples that were scanned, intracranial volume and skull length followed a similar pattern of increase up to P20 and then plateaued at older ages. These data are consistent with the possibility of ICP being a contributing factor to bone formation at the sutures during early stages of development. The data can be further used for development and validation of computational models of skull growth.

Activation of the IGF1 pathway mediates changes in cellular contractility and motility in single-suture craniosynostosis.

Insulin growth factor 1 (IGF1) is a major anabolic signal that is essential during skeletal development, cellular adhesion and migration. Recent transcriptomic studies have shown that there is an upregulation in IGF1 expression in calvarial osteoblasts derived from patients with single-suture craniosynostosis (SSC). Upregulation of the IGF1 signaling pathway is known to induce increased expression of a set of osteogenic markers that previously have been shown to be correlated with contractility and migration. Although the IGF1 signaling pathway has been implicated in SSC, a correlation between IGF1, contractility and migration has not yet been investigated. Here, we examined the effect of IGF1 activation in inducing cellular contractility and migration in SSC osteoblasts using micropost arrays and time-lapse microscopy. We observed that the contractile forces and migration speeds of SSC osteoblasts correlated with IGF1 expression. Moreover, both contractility and migration of SSC osteoblasts were directly affected by the interaction of IGF1 with IGF1 receptor (IGF1R). Our results suggest that IGF1 activity can provide valuable insight for phenotype-genotype correlation in SSC osteoblasts and might provide a target for therapeutic intervention.

Osteoblast differentiation profiles define sex specific gene expression patterns in craniosynostosis.

Single suture craniosynostosis (SSC) is the premature fusion of one calvarial suture and occurs in 1-1700-2500 live births. Congenital fusion of either the sagittal, metopic, or coronal sutures represents 95% of all cases of SSC. Sagittal and metopic synostosis have a male preponderance (3:1) while premature fusion of the coronal suture has a female preponderance (2:1). Although environmental and genetic factors contribute to SSC, the etiology of the majority of SSC cases remains unclear. In this study, 227 primary calvarial osteoblast cell lines from patients with coronal, metopic, or sagittal synostosis and unaffected controls were established and assayed for ALP activity and BrdU incorporation (n = 226) as respective measures of early stage osteoblast differentiation and proliferation. Primary osteoblast cell lines from individuals with sagittal synostosis demonstrated higher levels of ALP activity and reduced proliferation when compared to control lines. In order to address the sex differences in SSC types, the data was further stratified by sex. Osteoblasts from males and females with sagittal synostosis as well as males with metopic synostosis demonstrated higher levels of ALP activity when compared to sex matched controls, and males with sagittal or metopic synostosis demonstrated reduced levels of proliferation. In order to elucidate genes and pathways involved in these observed phenotypes, correlation analyses comparing ALP activity and proliferation to global gene expression was performed. Transcripts related to osteoblast differentiation were identified both differentially up and downregulated, correlated with ALP activity when compared to controls, and demonstrated a striking sex specific gene expression pattern. These data support that the dysregulation of osteoblast differentiation plays a role in the development of SSC and that genetic factors contribute to the observed sex related differences.

High-oleic rapeseed (canola) and flaxseed oils modulate serum lipids and inflammatory biomarkers in hypercholesterolaemic subjects.

Recently, novel dietary oils with modified fatty acid profiles have been manufactured to improve fatty acid intakes and reduce CVD risk. Our objective was to evaluate the efficacy of novel high-oleic rapeseed (canola) oil (HOCO), alone or blended with flaxseed oil (FXCO), on circulating lipids and inflammatory biomarkers v. a typical Western diet (WD). Using a randomised, controlled, crossover trial, thirty-six hypercholesterolaemic subjects consumed three isoenergetic diets for 28 d each containing approximately 36% energy from fat, of which 70% was provided by HOCO, FXCO or WD. Dietary fat content of SFA, MUFA, PUFA n-6 and n-3 was 6, 23, 5, 1% energy for HOCO; 6, 16, 5, 7·5% energy for FXCO; 11·5, 16, 6, 0·5% energy for WD. After 28 d, compared with WD, LDL-cholesterol was reduced 15·1% (P < 0·001) with FXCO and 7·4% (P < 0·001) with HOCO. Total cholesterol (TC) was reduced 11% (P < 0·001) with FXCO and 3·5% (P = 0·002) with HOCO compared with WD. Endpoint TC differed between FXCO and HOCO (P < 0·05). FXCO consumption reduced HDL-cholesterol by 8·5% (P < 0·001) and LDL:HDL ratio by 7·5% (P = 0·008) v. WD. FXCO significantly decreased E-selectin concentration compared with WD (P = 0·02). No differences were observed in inflammatory markers after the consumption of HOCO compared with WD. In conclusion, consumption of novel HOCO alone or when blended with flaxseed oil is cardioprotective through lipid-lowering effects. The incorporation of flaxseed oil may also target inflammation by reducing plasma E-selectin.

Economic valuation of the potential health benefits from foods enriched with plant sterols in Canada.

BACKGROUND: Increased consumption of foods containing plant sterols has the potential to reduce the incidence of coronary heart disease (CHD) and thus reduce costs associated with treating that disease in a significant way. This paper reports the results of an investigation of the potential monetary benefits of allowing foods enriched with plant sterols to be marketed in Canada. OBJECTIVE: The objective of this research was to estimate the annual savings that would accrue to Canada's single-payer publicly funded health care system if plant sterols were approved for use. If foods containing plant sterols are consumed at a sufficient rate, a reduction in CHD should follow. Given the significant costs associated with CHD, approval of plant sterols in Canada has important public policy implications. DESIGN: This research employs a variation of traditional cost-of-illness analysis entailing four steps: (1) estimation of a 'success rate' (proportion of persons who would consume plant sterols at the necessary rate); (2) presumption of blood cholesterol reduction due to plant sterol consumption; (3) assumption of reduction in CHD that follows from blood cholesterol reduction; and (4) calculation of cost savings associated with reduced incidence of CHD. RESULTS: CALCULATIONS WERE CARRIED OUT FOR FOUR SCENARIOS: ideal, optimistic, pessimistic, and very pessimistic. It was estimated that between $38 million (very pessimistic scenario) and $2.45 billion (ideal scenario) could be saved annually by Canada's health care system with plant sterol-enriched food products being made available for sale. CONCLUSION: Significant expenditure reductions within Canada's publicly funded health care system could be realized with plant sterols approved for sale. Reduced CHD resulting from lower blood cholesterol levels would lessen the financial burden of disease in Canada.

The stability of metabolic syndrome in children and adolescents.

CONTEXT: Some studies suggest the presence of metabolic syndrome before adulthood may identify those at high risk for later cardiovascular morbidity, but there are few data examining the reliability of pediatric metabolic syndrome. OBJECTIVE: To examine the short- and long-term stability of pediatric metabolic syndrome. DESIGN: Metabolic syndrome was defined as having at least three of the following: waist circumference, blood pressure, and fasting serum triglycerides in the 90th or higher percentile for age/sex; high-density lipoprotein-cholesterol 10th or lower percentile for age/sex; and fasting serum glucose of at least 100 mg/dl. Short-term metabolic syndrome stability (repeated measurements within 60 d) was assessed in obese youth ages 6-17 yr. Long-term metabolic syndrome stability (repeated measurements more than 1.5 yr apart) was studied in 146 obese and nonobese children age 6-12 yr at baseline. PATIENTS AND SETTING: Convenience samples of obese and nonobese youth ages 6-17 yr participating in research studies were collected at a clinical research hospital. RESULTS: Short-term metabolic syndrome stability (repeat measurements performed 19.7 +/- 13.1 d apart) was assessed in 220 children. The diagnosis of metabolic syndrome was unstable in 31.6% of cases. At their short-term follow-up visit, incidence of metabolic syndrome among participants who did not have metabolic syndrome at baseline was 24%. In the long term (repeat measurements performed 5.6 +/- 1.9 yr apart), the diagnosis of metabolic syndrome was unstable in 45.5% of cases. CONCLUSIONS: Cutoff-point-based definitions for pediatric metabolic syndrome have substantial instability in the short and long term. The value of making a cutoff-point-based diagnosis of metabolic syndrome during childhood or adolescence remains in question.

A prospective study of loss of control eating for body weight gain in children at high risk for adult obesity.

OBJECTIVE: Limited data suggest that disordered-eating may predispose children to excessive weight gain. We investigated the relationship between baseline responses to the Eating Disorder Examination adapted for Children (ChEDE) and change in BMI (kg/m(2)) in children at high risk for adult obesity. METHOD: Children (6-12 years) were administered the ChEDE to assess loss of control (LOC) eating, dietary restraint, and eating, shape, and weight concern. Height and weight were measured at baseline and annually. RESULTS: Between July, 1999, and August, 2007, 772 measurements were obtained from 143 children over 4.5 +/- 1.9 years. LOC eating predicted an increased rate of BMI growth over time (p = .02). Compared with children without LOC, those reporting LOC gained an additional mean 2.4 kg of weight per year. CONCLUSION: LOC is a salient predictor of weight gain during middle childhood. Interventions that decrease LOC eating should be evaluated for their ability to prevent excessive pediatric weight gain.

Insulin resistance, hyperinsulinemia, and energy intake in overweight children.

OBJECTIVE: To examine the relationship between energy intake during a buffet meal and indexes of insulin dynamics in overweight children. STUDY DESIGN: Ninety-five nondiabetic, overweight (body mass index > or = 95th percentile) children (age 10.3 +/- 1.4 years) selected lunch from a 9835-kcal buffet eaten ad libitum after an overnight fast. The associations between energy intake and measures of insulin dynamics, in the postabsorptive state and during a 2-hour hyperglycemic clamp, were determined. Covariates in the statistical model included race, sex, skeletal age, fat-free mass, fat mass, socioeconomic status, and number of foods in the buffet rated as acceptable. RESULTS: Energy intake was positively associated with the fasting homeostasis model assessment for insulin resistance index (beta = 0.24, P = .042), fasting insulin/glucose ratio (beta = 0.24, P = .044), first-phase insulin (beta = 0.23, P = .032), and first-phase C-peptide (beta = 0.21, P = .046); energy intake was negatively associated with clamp-derived insulin sensitivity (beta = -0.29, P = .042). Each 10% decrease in clamp-derived insulin sensitivity predicted a 27-kcal greater energy intake. CONCLUSIONS: Insulin resistance and hyperinsulinemia are associated with greater energy intake after an overnight fast in overweight children. These associations suggest mechanisms whereby insulin resistance may contribute to excessive weight gain in children.

Psychometric properties of a new questionnaire to assess eating in the absence of hunger in children and adolescents.

BACKGROUND: Eating in the absence of hunger (EAH), studied in the context of laboratory paradigms, has been associated with obesity and is predictive of excess weight gain in children. However, no easily administered questionnaire exists to assess for EAH in children. OBJECTIVE: We developed an Eating in the Absence of Hunger Questionnaire to be administered to children and adolescents (EAH-C) and examined psychometric properties of the measure. DESIGN: Two-hundred and twenty-six obese (BMI > or = 95th percentile for age and sex, n=73) and non-obese (BMI<95th percentile, n=153) youth (mean age+/-S.D., 14.4+/-2.5 y) completed the EAH-C and measures of loss of control and emotional eating, and general psychopathology. Temporal stability was assessed in a subset of participants. RESULTS: Factor analysis generated three subscales for the EAH-C: Negative Affect, External Eating, and Fatigue/Boredom. Internal consistency for all subscales was established (Cronbach's alphas: 0.80-0.88). The EAH-C subscales had good convergent validity with emotional eating and loss of control episodes (p's<0.01). Obese children reported higher Negative Affect subscale scores than non-obese children (p

Structure analysis of the Children's Eating Attitudes Test in overweight and at-risk for overweight children and adolescents.

BACKGROUND: In school-based samples of children, the Children's Eating Attitudes Test (ChEAT) has a four-factor structure; however, previous studies have not examined its factor structure in samples restricted to overweight youth. METHODS: The ChEAT was administered to 220 overweight (BMI>or=95th percentile) and 45 at-risk for overweight (BMI 85th-<95th percentile) children and adolescents. Factors were identified by a principal component analysis with varimax rotation. ChEAT factor scores of children with BMI>or=85th percentile were contrasted with those of 152 non-overweight (BMI 5th to <85th percentile) children and adolescents. RESULTS: Factor analysis generated four subscales described as 'body/weight concern,' 'food preoccupation,' 'dieting,' and 'eating concern.' ChEAT total score, body/weight concern, and dieting subscale scores were positively related to BMI-Z and body fat mass (p's<.05). Compared to non-overweight children, overweight and at-risk for overweight children had higher ChEAT total (9.9+/-7.4 vs. 6.6+/-7.8, p<.001), body/weight concern (3.2+/-3.1 vs. 1.3+/-3.0, p<.001), and dieting (1.8+/-2.2 vs. .8+/-2.3, p<.001) subscale scores. CONCLUSIONS: The previously elucidated factor structure of the ChEAT was primarily supported in a sample of overweight children. The emergence of separate body/weight concern and dieting subscales may relate to these children's experiences with attempted weight reduction.

Sigma1 and sigma2 receptor binding affinity and selectivity of SA4503 and fluoroethyl SA4503.

SA4503, a potent sigma(1) receptor agonist, is reported as having 103-fold higher affinity for sigma(1) (IC(50) = 17.4 nM) than sigma(2) (IC(50) = 1,784 nM) sites in guinea pig brain membranes. Modest structural changes appear to have major effects on sigma(1)/sigma(2) selectivity. The fluoroethyl analog, FE-SA4503, is described as having high primary affinity for sigma(2) sites (IC(50) = 2.11 nM) and a weaker interaction with sigma(1) sites (IC(50) = 6.48 nM). SA4503 and FE-SA4503 have been radiolabeled for PET studies, and both bind selectively to sigma(1) receptors in animal and human brain in vivo. We prepared SA4503 and FE-SA4503 as reference compounds for radioligand development efforts. In our hands, SA4503 is 14-fold selective for sigma(1) (K(i) = 4.6 nM) over sigma(2) (K(i) = 63.1 nM) sites in guinea pig brain homogenates. Further, FE-SA4503 exhibits the same 14-fold selectivity for sigma(1) (K(i) = 8.0 nM) over sigma(2) (K(i) = 113.2 nM) receptors. The main differences from previously reported values stem from sigma(2) affinity determinations. This protocol, displacement of [(3)H]DTG binding to sigma(2) sites using (+)-pentazocine (200 nM) to mask sigma(1) sites, was validated by the proper rank order of sigma(2) inhibitory potencies shown by a panel of additional ligands: ifenprodil > haloperidol > DTG >> (+)-pentazocine. Robust Pearson correlation (r = 1.0, P = 0.002; slope = 0.97) was observed for our pK(i) values against those from a prior study by others. The findings have bearing on structure-activity relationships for this active series, and on conclusions that might be drawn from experiments relying upon defined sigma(1)/sigma(2) binding selectivity.

Stability of a PKCI-1-related mRNA is controlled by the splicing factor ASF/SF2: a novel function for SR proteins.

Pre-mRNA splicing is a widely used regulatory mechanism for controlling gene expression, and a family of conserved proteins, SR proteins, participate in both constitutive and alternative splicing. Here we describe a novel function for the SR protein ASF/SF2. We used an embryonic chicken cDNA library to screen for differential mRNA expression in the chicken B-cell line DT40-ASF, expressing or not expressing ASF/SF2. Remarkably, out of 3 x 10(6) clones screened, only one, isolated several times independently, showed ASF/SF2-related differential expression. The isolated cDNA, referred to here as PKCI-r (for PKCI-related), is closely related to the protein kinase C interacting protein (PKCI-1) gene. Transcript levels were increased approximately sixfold in ASF/SF2-depleted cells compared with cells expressing ASF/SF2, indicating a negative role for the SR protein. Strikingly, inhibition of ASF/SF2 expression had no significant effect on PKCI-r splicing, or transcription, but markedly increased the half-life of PKCI-r mRNA (6.6-fold). Similarly, increased mRNA stability was also observed upon expression of exogenous PKCI-r mRNA in cells depleted of ASF/SF2. ASF/SF2 bound to a discrete region containing a purine-rich sequence in the 3' UTR of the PKCI-r transcript, and deletion of this region eliminated ASF/SF2-mediated regulation of transcript stability. Together these data indicate a novel, direct effect of ASF/SF2 on PKCI-r mRNA stability. Therefore, ASF/SF2, and perhaps other SR proteins, affects gene expression in vertebrate cells through regulation of mRNA stability as well as splicing.