Functional connectivity during facial and vocal emotion recognition: Preliminary evidence for dissociations in developmental change by nonverbal modality.

The developmental trajectory of emotion recognition (ER) skills is thought to vary by nonverbal modality, with vocal ER becoming mature later than facial ER. To investigate potential neural mechanisms contributing to this dissociation at a behavioural level, the current study examined whether youth's neural functional connectivity during vocal and facial ER tasks showed differential developmental change across time. Youth ages 8-19 (n = 41) completed facial and vocal ER tasks while undergoing functional magnetic resonance imaging, at two timepoints (1 year apart; n = 36 for behavioural data, n = 28 for neural data). Partial least squares analyses revealed that functional connectivity during ER is both distinguishable by modality (with different patterns of connectivity for facial vs. vocal ER) and across time-with changes in connectivity being particularly pronounced for vocal ER. ER accuracy was greater for faces than voices, and positively associated with age; although task performance did not change appreciably across a 1-year period, changes in latent functional connectivity patterns across time predicted participants' ER accuracy at Time 2. Taken together, these results suggest that vocal and facial ER are supported by distinguishable neural correlates that may undergo different developmental trajectories. Our findings are also preliminary evidence that changes in network integration may support the development of ER skills in childhood and adolescence.

Proactive Versus Reactive Control Strategies Differentially Mediate Alcohol Drinking in Male Wistars and P Rats.

Problematic alcohol consumption is associated with deficits in decision-making and alterations in prefrontal cortex neural activity likely contribute. We hypothesized that the differences in cognitive control would be evident between male Wistars and a model of genetic risk: alcohol-preferring P rats. Cognitive control is split into proactive and reactive components. Proactive control maintains goal-directed behavior independent of a stimulus, whereas reactive control elicits goal-directed behavior at the time of a stimulus. We hypothesized that Wistars would show proactive control over alcohol seeking whereas P rats would show reactive control over alcohol seeking. Neural activity was recorded from the prefrontal cortex during an alcohol seeking task with two session types. On congruent sessions, the conditioned stimulus (CS+) was on the same side as alcohol access. Incongruent sessions presented alcohol opposite the CS+. Wistars, but not P rats, made more incorrect approaches during incongruent sessions, suggesting that Wistars utilized the previously learned rule. This motivated the hypothesis that neural activity reflecting proactive control would be observable in Wistars but not P rats. While P rats showed differences in neural activity at times of alcohol access, Wistars showed differences prior to approaching the sipper. These results support our hypothesis that Wistars are more likely to engage in proactive cognitive control strategies whereas P rats are more likely to engage in reactive cognitive control strategies. Although P rats were bred to prefer alcohol, the differences in cognitive control may reflect a sequela of behaviors that mirror those in humans at risk for an AUD.

Proactive Versus Reactive Control Strategies Differentially Mediate Alcohol Drinking in Wistar and P rats.

Problematic alcohol consumption is associated with deficits in decision-making, and alterations in prefrontal cortex neural activity likely contributes. We hypothesized that differences in cognitive control would be evident between male Wistar rats and a model for genetic risk for alcohol use disorder (alcohol-preferring P rats). Cognitive control can be split into proactive and reactive components. Proactive control maintains goal-directed behavior independent of a stimulus whereas reactive control elicits goal-directed behavior at the time of a stimulus. We hypothesized that Wistars would show proactive control over alcohol-seeking whereas P rats would show reactive control over alcohol-seeking. Neural ensembles were recorded from prefrontal cortex during an alcohol seeking task that utilized two session types. On congruent sessions the CS+ was on the same side as alcohol access. Incongruent sessions presented alcohol opposite the CS+. Wistars, but not P rats, exhibited an increase in incorrect approaches during incongruent sessions, suggesting that Wistars utilized the previously learned task-rule. This motivated the hypothesis that ensemble activity reflecting proactive control would be observable in Wistars but not P rats. While P rats showed differences in neural activity at times relevant for alcohol delivery, Wistars showed differences prior to approaching the sipper. These results support our hypothesis that Wistars are more likely to engage proactive cognitive-control strategies whereas P rats are more likely to engage reactive cognitive control strategies. Although P rats were bred to prefer alcohol, differences in cognitive control may reflect a sequela of behaviors that mirror those in humans at risk for an AUD.

Functional patterns of neural activation during vocal emotion recognition in youth with and without refractory epilepsy.

Epilepsy has been associated with deficits in the social cognitive ability to decode others' nonverbal cues to infer their emotional intent (emotion recognition). Studies have begun to identify potential neural correlates of these deficits, but have focused primarily on one type of nonverbal cue (facial expressions) to the detriment of other crucial social signals that inform the tenor of social interactions (e.g., tone of voice). Less is known about how individuals with epilepsy process these forms of social stimuli, with a particular gap in knowledge about representation of vocal cues in the developing brain. The current study compared vocal emotion recognition skills and functional patterns of neural activation to emotional voices in youth with and without refractory focal epilepsy. We made novel use of inter-subject pattern analysis to determine brain areas in which activation to emotional voices was predictive of epilepsy status. Results indicated that youth with epilepsy were comparatively less able to infer emotional intent in vocal expressions than their typically developing peers. Activation to vocal emotional expressions in regions of the mentalizing and/or default mode network (e.g., right temporo-parietal junction, right hippocampus, right medial prefrontal cortex, among others) differentiated youth with and without epilepsy. These results are consistent with emerging evidence that pediatric epilepsy is associated with altered function in neural networks subserving social cognitive abilities. Our results contribute to ongoing efforts to understand the neural markers of social cognitive deficits in pediatric epilepsy, in order to better tailor and funnel interventions to this group of youth at risk for poor social outcomes.

Social brain networks: Resting-state and task-based connectivity in youth with and without epilepsy.

Individuals with epilepsy often experience social difficulties and deficits in social cognition. It remains unknown how disruptions to neural networks underlying such skills may contribute to this clinical phenotype. The current study compared the organization of relevant brain circuits-the "mentalizing network" and a salience-related network centered on the amygdala-in youth with and without epilepsy. Functional connectivity between the nodes of these networks was assessed, both at rest and during engagement in a social cognitive task (facial emotion recognition), using functional magnetic resonance imaging. There were no group differences in resting-state connectivity within either neural network. In contrast, youth with epilepsy showed comparatively lower connectivity between the left posterior superior temporal sulcus and the medial prefrontal cortex-but greater connectivity within the left temporal lobe-when viewing faces in the task. These findings suggest that the organization of a mentalizing network underpinning social cognition may be disrupted in youth with epilepsy, though differences in connectivity within this circuit may shift depending on task demands. Our results highlight the importance of considering functional task-based engagement of neural systems in characterizations of network dysfunction in epilepsy.

Children and adolescents' neural response to emotional faces and voices: Age-related changes in common regions of activation.

The perception of facial and vocal emotional expressions engages overlapping regions of the brain. However, at a behavioral level, the ability to recognize the intended emotion in both types of nonverbal cues follows a divergent developmental trajectory throughout childhood and adolescence. The current study a) identified regions of common neural activation to facial and vocal stimuli in 8- to 19-year-old typically-developing adolescents, and b) examined age-related changes in blood-oxygen-level dependent (BOLD) response within these areas. Both modalities elicited activation in an overlapping network of subcortical regions (insula, thalamus, dorsal striatum), visual-motor association areas, prefrontal regions (inferior frontal cortex, dorsomedial prefrontal cortex), and the right superior temporal gyrus. Within these regions, increased age was associated with greater frontal activation to voices, but not faces. Results suggest that processing facial and vocal stimuli elicits activation in common areas of the brain in adolescents, but that age-related changes in response within these regions may vary by modality.

Use of alternative medicine among patients with head and neck cancer.

OBJECTIVES: To determine the prevalence of alternative medicine use in the population with head and neck cancer and correlate with demographics and tumor characteristics. DESIGN: Cross-sectional survey study. SETTING: Two tertiary cancer centers. PATIENTS: Two hundred consecutive outpatients with consecutive head and neck cancer. INTERVENTIONS: A 10- to 25-minute patient interview administered by primary investigator. MAIN OUTCOME MEASURES: Demographic markers (sex, age, education, household income, marital status, ethnic background, and geographic location); tumor characteristics (tumor site, pathology, staging, time since diagnosis, and incidence of recurrence); conventional mode of treatment; attitudes regarding alternative medicine, source of exposure to alternative medicine, therapeutic rationale, treatment efficacy, sources of information, and discussions with physicians about alternative medicine. RESULTS: Seventy-seven (38.5%) of 200 patients had used alternative medicine for some purpose, and 45 (22.5%) of 200 did so for head and neck cancer. Increased use of alternative medicine occurred among patients of younger age, having a postsecondary education, higher personal income, and Indo-Asian extraction. Of those patients using alternative anticancer therapy, increased use was noted among patients with tumors of the nasopharynx, nonsquamous cell carcinoma pathology, and recurrent disease. Conventional mode of treatment had no association with alternative medicine use. Physicians were believed to be the most knowledgeable about alternative medicine, while the usual proponents of alternative medicine were identified least frequently. CONCLUSIONS: Alternative cancer therapy use among patients with head and neck cancer was 22.5%, with increased use in younger, affluent, better educated patients, and those of Indo-Asian extraction. Patients view physicians as being knowledgeable about alternative medicine. Otolaryngologists should inform themselves about alternative medicine to counsel patients more effectively.

Specificity of mutagenesis by 4-aminobiphenyl: mutations at G residues in bacteriophage M13 DNA and G-->C transversions at a unique dG(8-ABP) lesion in single-stranded DNA.

Mutagenesis by the human bladder carcinogen 4-aminobiphenyl (ABP) was studied in single-stranded DNA from a bacteriophage M13 cloning vector. In comparison to ABP lesions in double-stranded DNA, lesions in single-stranded DNA were approximately 70-fold more mutagenic and 50-fold more genotoxic. Sequencing analysis of ABP-induced mutations in the lacZ gene revealed exclusively base-pair substitutions, with over 80% of the mutations occurring at G sites; the G at position 6310 accounted for 25% of the observed mutations. Among the sequence changes at G sites, G-->T transversions predominated, followed by G-->C transversions and G-->A transitions. In order to further elucidate the mutagenic mechanism of ABP, an oligonucleotide containing the major DNA adduct, N-(deoxyguanosin-8-yl)-4-aminobiphenyl (dG(8-ABP)), was situated within the PstI site of a single-stranded M13 genome. After in vivo replication of the adduct containing ABP-modified and control (unadducted) genomes, the mutational frequency and mutational specificity of the dG(8-ABP) lesion were determined. The targeted mutational efficiency was approximately 0.01%, and the primary mutation observed was the G-->C transversion. Thus dG(8-ABP), albeit weakly mutagenic at the PstI site, can contribute to the mutational spectrum of ABP lesions.

Synthesis of oligonucleotides containing two putatively mutagenic DNA lesions: 5-hydroxy-2'-deoxyuridine and 5-hydroxy-2'-deoxycytidine.

Spontaneous oxidative DNA damage occurs as a consequence of aerobic metabolism, lipid peroxidation, immune responses, ionizing radiation, and some chemical oxidants. These processes yield a vast array of oxidized DNA bases and sugars. The existence of significant steady-state levels of oxidized DNA bases in the genome suggests that these lesions are not completely repaired on a biologically relevant time scale and thus may contribute to mutagenesis. In particular, studies have shown that the steady-state levels of 5-hydroxy-2'-deoxycytidine (dC5-OH) and its deamination product, 5-hydroxy-2'-deoxyuridine (dU5-OH), are similar to those found for 7,8-dihydro-8-oxoguanosine, a known highly mutagenic lesion formed by oxidation of guanosine. Structural and biological properties of dC5-OH and dU5-OH have been constrained by the lack of synthetic methodology for oligonucleotides containing these modified bases. A method is described here for the solid-phase synthesis of oligonucleotides containing dC5-OH and dU5-OH. Preparation of each of the required phosphoramidites involved the selective protection of the base 5-hydroxyl group over the deoxyribose 5'- and 3'-hydroxyl groups. The base composition and the incorporation of the adducts into synthetic heptanucleotides were confirmed after purification of the modified oligonucleotides by enzymatic digestion and HPLC analysis. Mass spectrometric analysis of the oligonucleotide products by electrospray MS and GC/MS further confirmed their composition. Most significantly, deamination of the dC5-OH oligomer to a putative dU5-OH product during solid-phase DNA synthesis or oligonucleotide deprotection was not detected by any analytical technique employed.

Synthesis and biological activity of novel nonnucleoside inhibitors of HIV-1 reverse transcriptase. 2-Aryl-substituted benzimidazoles.

The development of new nonnucleoside inhibitors of human immunodeficiency virus type-1 (HIV-1) reverse transcriptase (RT) active against the drug-induced mutations in RT continues to be a very important goal of AIDS research. We used a known inhibitor of HIV-1 RT, 1-(2,6-difluorophenyl)-1H,3H-thiazolo[3,4-alpha]benzimidazole (TZB), as the lead structure for drug design with the objective of making more potent inhibitors against both wild-type (WT) and variant RTs. A series of structurally related 1,2-substituted benzimidazoles was synthesized and evaluated for their ability to inhibit in vitro polymerization by HIV-1 WT RT. A structure-activity study was carried out for the series of compounds to determine the optimum groups for substitution of the benzimidazole ring at the N1 and C2 positions. The best inhibitor, 1-(2,6-difluorobenzyl)-2-(2,6-difluorophenyl)-4-methylbenzimida zole (35), has an IC50 = 200 nM against HIV-1 WT RT in an in vitro enzyme assay. Cytoprotection assays utilizing HIV-infected MT-4 cells revealed that 35 had strong antiviral activity (EC50 = 440 nM) against wild-type virus while retaining broad activity against many clinically observed HIV-1 strains resistant to nonnucleoside inhibitors. Overall, the activity of 35 against wild-type and resistant strains with amino acid substitution in RT is 4-fold or greater than that of TZB and is comparable to that of other nonnucleoside inhibitors currently undergoing clinical trials, most of which do not have the capacity to inhibit the variant forms of the enzyme.

X-ray structure of a DNA decamer containing 7,8-dihydro-8-oxoguanine.

We have determined the x-ray structure of a DNA fragment containing 7,8-dihydro-8-oxoguanine (G(O)). The structure of the duplex form of d(CCAGOCGCTGG) has been determined to 1.6-A resolution. The results demonstrate that GO forms Watson-Crick base pairs with the opposite C and that G(O) is in the anti conformation. Structural perturbations induced by C.G(O)anti base pairs are subtle. The structure allows us to identify probable elements by which the DNA repair protein MutM recognizes its substrates. Hydrogen bond donors/acceptors within the major groove are the most likely element. In that groove, the pattern of hydrogen-bond donors/acceptors of C.G(O)anti is unique. Additional structural analysis indicates that conversion of G to G(O) would not significantly influence the glycosidic torsion preference of the nucleoside. There is no steric interaction of the 8-oxygen of G(O) with the phospho-deoxyribose backbone.

Peracylation of nucleosides with methionine: foundation for a method to detect carcinogen adducts.

We report the chemical foundation for a new method to detect carcinogen-DNA adducts, which we have designated adduct detection by acylation with methionine (ADAM). The method is based on reaction of DNA adducts with a protected methionine derivative, (tert-butoxycarbonyl)-L-methionine N-hydroxysuccinimidyl ester (TMB-NHS). Acylation of 2'-deoxyguanosine (dGuo), used as a prototypical deoxynucleoside, and N-(deoxyguanosin-8-yl)-4-aminobiphenyl (dGuo-8-ABP), the major DNA adduct formed after in vivo exposure to 4-aminobiphenyl, a known human carcinogen, with TBM-NHS was optimized, and products were characterized by 3H radioactivity, UV absorbance, mass spectrometry, and 1H and 13C NMR. Derivatives acylated on hydroxyl (5' or 3') and/or amine (N2) groups were unambiguously determined to be mono-, bis-, and tris-TBM-acylated nucleosides. Under optimal acylation conditions [TBM-NHS (> or = 4 x 10(5) molar equivalents), pyridine (50 microL), THF (50 microL), and diisopropylcarbodiimide (DIC) (1 microL) and incubation for 2 h at 37 degrees C], the efficiency of acylation for picomole or smaller quantities of dGuo-8-ABP exceeded 95%, with the tris-TMB-acylated nucleoside representing the major product (88%). A linear correlation was obtained between the amount of [3H]dGuo-8-ABP introduced into the reaction and the total amount of TBM-acylated products formed. These results support the validity of this strategy for adaptation as an analytical method for the detection of low levels of DNA adducts through the use of (tert-butoxycarbonyl)-L-[35S]-methionine N-hydroxysuccinimidyl ester.

Adduct detection by acylation with [35S]methionine: analysis of DNA adducts of 4-aminobiphenyl.

Reaction of synthetic N-(2'-deoxyguanosin-8-yl)-4-aminobiphenyl (dGuo-8-ABP) with t-butoxycarbonyl-L-[35S]methionine, N-hydroxysuccinimidyl ester (35S-labeled TBM-NHS), under optimized conditions produced mono-, bis-, and tris-TBM-acylated nucleosides that were separable by HPLC. Reaction of different amounts of N-(2'-deoxy-1',2'-[3H]guanosin-8-yl)-4-aminobiphenyl ([3H]dGuo-8-ABP) with 35S-labeled TBM-NHS established that total 35S content of acylated products was linearly related to adduct concentration (r = 0.992) over the range of 10 fmol to 30.6 pmol. Additionally, the N-(deoxyguanosin-8-yl)-4-[3H]aminobiphenyl (dGuo-8-[3H]ABP) adduct was isolated from calf thymus DNA adducted in vitro and from rat liver DNA adducted in vivo and similarly reacted with 35S-labeled TBM-NHS. Acylation products of dGuo-8-ABP from all three sources showed HPLC retention times identical to those of authentic TBM-dGuo-8-ABP, and 35S incorporation into acylated products was linearly related to amount of adduct reacted. These results indicate that the procedure, to which we have referred as adduct detection by acylation with methionine (ADAM), has potential applicability as an analytical procedure for detection and quantification of DNA adducts in human tissues in the molecular epidemiology of cancer.

Genetic effects of oxidative DNA damage: comparative mutagenesis of 7,8-dihydro-8-oxoguanine and 7,8-dihydro-8-oxoadenine in Escherichia coli.

A single 7,8-dihydro-8-oxoguanine (G8-OXO; 8-hydroxyguanine) adduct in the lacZ alpha gene of bacteriophage M13 DNA induces a targeted G-->T transversion after replication in Escherichia coli (Biochemistry, 29, 7024-7031 (1990)). This mutation is thought to be due to the facile formation during DNA synthesis of a G8-OXO.base pair, where G8-OXO is in the syn conformation about the deoxyglycosyl bond. A related modified purine, 7,8-dihydro-8-oxoadenine (A8-OXO; 8-hydroxyadenine), is an abundant product found in irradiated and oxidized DNAs. Similar to G8-OXO, as a mononucleoside A8-OXO assumes the syn conformation. This work has assessed the relative mutagenicities of A8-OXO and G8-OXO in the same experimental system. A deoxypentanucleotide containing A8-OXO [d(GCT-A8-OXOG)] was synthesized. After 5'-phosphorylation with [gamma-32P] ATP, the oligomer was ligated into a duplex M13mp19-derived genome at a unique NheI restriction site. Genomes containing either A8-OXO (at position 6275, [+] strand) or G8-OXO (position 6276) were denatured with heat and introduced into E.coli DL7 cells. Analysis of phage DNA from mutant plaques obtained by plating immediately after transformation (infective centers assay) revealed that G8-OXO induced G-->T transversions at an apparent frequency of approximately 0.3%. The frequency and spectrum of mutations observed in DNA sequences derived from 172 mutant plaques arising from the A8-OXO-modified DNA were almost indistiguishable from those generated from transfection of an adenine-containing control genome. We conclude that A8-OXO is at least an order of magnitude less mutagenic than G8-OXO in E.coli cells with normal DNA repair capabilities.

Generation and characterization of monoclonal antibodies to prostate secretory protein.

Monoclonal antibodies (MAbs) were produced against a highly purified preparation of prostate secretory protein (PSP) isolated from normal seminal plasma. Fifteen antibodies were selected for further evaluation based on their strong reactivity and specificity for PSP. All the MAbs had a specificity for prostate epithelial cells and none reacted to any of a variety of normal tissues as determined by immunoperoxidase staining. Six of the MAbs were selected for further immunohistochemical evaluation based on their ability to recognize different antigenic determinants. Using competitive binding immunoassays, a variety of overlapping specificities were observed with at least 2 distinct epitopes identified. Although some staining variability was noted, the 6 antibodies, in general, gave the same pattern of tissue reactivity. Both the normal prostate and the benign prostate hyperplastic ductal epithelial cells stained intensely, with 78 to 100% and 50-100% of the cells staining, respectively. The number and often the staining intensity of the tumor cells decreased as the tumor became more undifferentiated. Approximately 40 to 100% and 15 to 70% of the tumor cells stained in the moderately-differentiated and well-differentiated carcinoma tissues, respectively, whereas either no staining was observed or less than 20% of the tumor cells stained in the poorly-differentiated and undifferentiated tumors. Most of the metastatic prostate tumors showed either no staining or scattered staining in a few cells (i.e., less than 20%).