Summary Objective We aimed to predict language deficits after epilepsy surgery. In addition to evaluating surgical factors examined previously, we determined the impact of the extent of functional magnetic resonance imaging (fMRI) activation that was resected on naming ability. Method Thirty‐five adults (mean age 37.5 ± 10.9 years, 13 male) with temporal lobe epilepsy completed a preoperative fMRI auditory description decision task, which reliably activates frontal and temporal language networks. Patients underwent temporal lobe resections (20 left resection). The Boston Naming Test (BNT) was used to determine language functioning before and after surgery. Language dominance was determined for Broca and Wernicke area (WA) by calculating a laterality index following statistical parametric mapping processing. We used an innovative method to generate anatomic resection masks automatically from pre‐ and postoperative MRI tissue map comparison. This mask provided the following: (a) resection volume; (b) overlap between resection and preoperative activation; and (c) overlap between resection and WA. We examined postoperative language change predictors using stepwise linear regression. Predictors included parameters described above as well as age at seizure onset (ASO), preoperative BNT score, and resection side and its relationship to language dominance. Results Seven of 35 adults had significant naming decline (6 dominant‐side resections). The final regression model predicted 38% of the naming score change variance (adjusted r2 = 0.28, P = 0.012). The percentage of top 10% fMRI activation resected (P = 0.017) was the most significant contributor. Other factors in the model included WA LI, ASO, volume of WA resected, and WA LI absolute value (extent of laterality). Significance Resection of fMRI activation during a word‐definition decision task is an important factor for postoperative change in naming ability, along with other previously reported predictors. Currently, many centers establish language dominance using fMRI. Our results suggest that the amount of the top 10% of language fMRI activation in the intended resection area provides additional predictive power and should be considered when planning surgical resection.

Infants born to women with depressive symptoms are at higher risk for insecure attachment and behavioral problems. Thus current medical practice is to continue psychotropic medication of pregnant women with depression despite concerns about its behavioral teratology. There are few animal studies focused on long-term behavioral effects of prenatal antidepressant exposure; in addition, studies have not looked at individual differences in baseline affective state as a source of response variability. In this study, fluoxetine, a selective serotonin reuptake inhibitor (SSRI), was administered to male and female rat pups from postnatal days 2-7 to model exposure to antidepressants in the human third trimester. Four behavioral measures were conducted from the neonatal to adult age periods in Low and High lines selectively bred for their rate of ultrasonic vocalizations after brief maternal separation. Neonatal fluoxetine administration decreased distress calls in both lines, but to a greater extent in High line rats than Low line. Neonatal fluoxetine also impaired motor coordination in neonates. Neonatal fluoxetine administration decreased social behavior in both juvenile and adult subjects. Fluoxetine-related reductions in anxiety behavior were not observed at the two older ages. As expected, High line subjects displayed more anxiety behavior than Low line subjects at all three test ages. These results suggest that there are may be significant behavioral consequences of antidepressant use during late pregnancy on offspring maternal attachment and social behavior, with implications for increased risk of autism spectrum disorders.

Objective To compare manual and automated pre- and postoperative hippocampal volume measurements in patients with intractable epilepsy. Methods We studied 34 patients referred to the Clinical Epilepsy Section, NINDS, NIH for evaluation of intractable epilepsy and 21 normal volunteers who received 1.5 or 3 T GE Signa MRI scans. Hippocampal volumes were manually traced on each slice and assembled into three-dimensional volumes by investigators blinded to other data. Automated volumetric measurements were obtained using FreeSurfer (http://surfer.nmr.mgh.harvard.edu/). Statistical analysis was performed with GraphPad Prism. Results Automated hippocampal volumes were larger than manual volumes in both patients and normal volunteers, p<.05. Right to left hemisphere hippocampal ratio and percent of hippocampus resected did not significantly differ by segmentation method. It was not possible to obtain accurate total resection volumes with the automated method. Significance Values such as side-to-side ratio and percent resected may be more directly translatable between manual and automated methods than absolute measures of volume. Accurate determination of resection volumes is important for studies of the effects of surgery on both seizure control and postoperative neuropsychological deficits. Our preliminary data suggest that FreeSurfer may provide an accurate and simple method for quantitating hippocampal resections. However, it may be less valuable for large or extratemporal resections, or when distortions of normal anatomy are present.

Phosphatidylinositol (4,5)-bisphosphate (PI(4,5)P2) is an important component of the inner leaflet of the plasma membrane of eukaryotic cells. Despite the fact that it only comprises approximately 1 mol% of the total membrane phospholipids, this phosphoinosite has been associated with many different cells functions including membrane trafficking, actin cytoskeleton remodeling and cell motility, among others. The physiological functions of this lipid seem to depend on localized concentration fluctuations within the plasma membrane. In fact, the distribution of this lipid in the plasma membrane has been proposed to be heterogeneous, and PI(4,5)P2 clustering is detected on model membranes under specific conditions. Domains highly enriched in PI(4,5)P2 were also reported at the plasma membrane of specific cell types. However, for most cellular models, scarce evidence has been found for PI(4,5)P2 segregation/clustering in the plasma membrane. In this context, our main goal was to study the distribution of PI(4,5)P2 molecules in cells lines where no heterogeneity in PI(4,5)P2 lateral distribution had been previously observed. The distribution of PI(4,5)P2 was assessed from FRET microscopy measurements with pleckstrin homology (PH) domains tagged with different fluorescent proteins. We applied a FRET methodology capable of discriminating between FRET from aggregates/clusters from noninteracting molecules within the plasma membrane of living cells. Our results clearly show distinct PI(4,5)P2 local densities in different cellular models, suggesting different patterns of PI(4,5)P2 lateral distributions within the plasma membrane. In addition, the effect of cholesterol removal on PI(4,5)P2 lateral organization is significantly different in distinct cell lines, suggesting that the role of cholesterol in the formation of PI(4,5)P2 enriched domains varies considerably. The membrane spanning G-protein coupled receptors (GPCRs) facilitate crucial physiological responses to a variety of extracellular ligands, such as hormones, neurotransmitters, ions, photons, and other stimuli [1], thus 50% of drugs available in the market today target GPCRs [2]. The biological mechanisms behind GPCR-signaling involves several other biomolecules, including the key protein group of b-arrestins. These cytosolic adaptor proteins regulate signaling through several different pathways, classically by removing receptors from the plasma membrane via clathrin-mediated endocytosis [3]. In membrane binding experiments using both confocal fluorescence microscopy and quartz crystal microbalance, we found that b-arrestin-1 binds specifically to supported lipid bilayers (SLBs) containing phosphatidylinositol trisphosphate lipids, PI(3,4,5)P3. We have consistently shown, using both methods, that such binding is lipid specific and not driven by membrane charge. We have also shown that the membrane-protein interaction depends on protein concentration and membrane composition, and that b-arrestins can induce membrane curvature, which may play a role in the endocyt...

brightness. The statistical distribution of probe molecules per platform was determined by single molecule brightness analysis. For demonstration, we used the consensus raft marker glycosylphosphatidylinositol-anchored monomeric GFP and the fluorescent lipid analogue Bodipy-GM1 which preferentially partitions into liquid ordered phases. For both markers we found cholesterol-dependent homo-association in the plasma membrane of living CHO and Jurkat T-cells in the resting state, thereby demonstrating the existence of small, mobile, stable platforms containing these probes. To further validate our method we extended TOCCSL by utilizing two-color co-localization and photo-activation. While two-color TOCCSL allows for direct imaging of mobile nanoplatforms containing different probe molecules and thus supporting the lipid raft concept, photo-activation based TOCCSL addresses an additional population of observed probes. Since TOCCSL is suitable for characterizing the mobile fraction of marker-proteins/lipids, the information about slowly diffusing or immobile nanoplatforms is not accessible. This can be circumvented by substituting the fluorescent marker by a photoactivatable protein linked to the molecule of interest. By irreversibly switching a small fraction of markers from a dark into the fluorescent state single molecule brightness and diffusion analysis after activation will add to the characterization of nanodomains.