Backward masking reveals coarse-to-fine dynamics in human V1.

Natural images exhibit luminance variations aligned across a broad spectrum of spatial frequencies (SFs). It has been proposed that, at early stages of processing, the coarse signals carried by the low SF (LSF) of the visual input are sent rapidly from primary visual cortex (V1) to ventral, dorsal and frontal regions to form a coarse representation of the input, which is later sent back to V1 to guide the processing of fine-grained high SFs (i.e., HSF). We used functional resonance imaging (fMRI) to investigate the role of human V1 in the coarse-to-fine integration of visual input. We disrupted the processing of the coarse and fine content of full-spectrum human face stimuli via backward masking of selective SF ranges (LSFs: <1.75cpd and HSFs: >1.75cpd) at specific times (50, 83, 100 or 150 ms). In line with coarse-to-fine proposals, we found that (1) the selective masking of stimulus LSF disrupted V1 activity in the earliest time window, and progressively decreased in influence, while (2) an opposite trend was observed for the masking of stimulus' HSF. This pattern of activity was found in V1, as well as in ventral (i.e. the Fusiform Face area, FFA), dorsal and orbitofrontal regions. We additionally presented subjects with contrast negated stimuli. While contrast negation significantly reduced response amplitudes in the FFA, as well as coupling between FFA and V1, coarse-to-fine dynamics were not affected by this manipulation. The fact that V1 response dynamics to strictly identical stimulus sets differed depending on the masked scale adds to growing evidence that V1 role goes beyond the early and quasi-passive transmission of visual information to the rest of the brain. It instead indicates that V1 may yield a 'spatially registered common forum' or 'blackboard' that integrates top-down inferences with incoming visual signals through its recurrent interaction with high-level regions located in the inferotemporal, dorsal and frontal regions.

Selection and characterization of HIV-1 with a novel S68 deletion in reverse transcriptase.

Resistance to human immunodeficiency virus type 1 (HIV-1) represents a significant problem in the design of novel therapeutics and the management of treatment regimens in infected persons. Resistance profiles can be elucidated by defining modifications to the viral genome conferred upon exposure to novel nucleoside reverse transcriptase (RT) inhibitors (NRTI). In vitro testing of HIV-1LAI-infected primary human lymphocytes treated with ß-D-2',3'-dideoxy-2',3'-didehydro-5-fluorocytidine (DFC; Dexelvucitabine; Reverset) produced a novel deletion of AGT at codon 68 (S68Δ) alone and in combination with K65R that differentially affects drug response. Dual-approach clone techniques utilizing TOPO cloning and pyrosequencing confirmed the novel S68Δ in the HIV-1 genome. The S68Δ HIV-1 RT was phenotyped against various antiviral agents in a heteropolymeric DNA polymerase assay and in human lymphocytes. Drug susceptibility results indicate that the S68Δ displayed a 10- to 30-fold increase in resistance to DFC, lamivudine, emtricitabine, tenofovir, abacavir, and amdoxovir and modest resistance to stavudine, ß-d-2',3'-oxa-5-fluorocytidine, or 9-(ß-D-1,3-dioxolan-4-yl)guanine and remained susceptible to 3'-azido-3'-deoxythymidine, 2',3'-dideoxyinosine (ddI), 1-(ß-D-dioxolane)thymine (DOT) and lopinavir. Modeling revealed a central role for S68 in affecting conformation of the ß3-ß4 finger region and provides a rational for the selective resistance. These data indicate that the novel S68Δ is a previously unrecognized deletion that may represent an important factor in NRTI multidrug resistance treatment strategies.

Inhibition of hepatitis C replicon RNA synthesis by beta-D-2'-deoxy-2'-fluoro-2'-C-methylcytidine: a specific inhibitor of hepatitis C virus replication.

beta-D-2'-Deoxy-2'-fluoro-2'-C-methylcytidine (PSI-6130) is a cytidine analogue with potent and selective anti-hepatitis C virus (HCV) activity in the subgenomic HCV replicon assay, 90% effective concentration (EC90)=4.6 +/- 2.0 microM. The spectrum of activity and cytotoxicity profile of PSI-6130 was evaluated against a diverse panel of viruses and cell types, and against two additional HCV-1b replicons. The S282T mutation, which confers resistance to 2'-C-methyl adenosine and other 2'-methylated nucleosides, showed only a 6.5-fold increase in EC90. When assayed for activity against bovine diarrhoea virus (BVDV), which is typically used as a surrogate assay to identify compounds active against HCV, PSI-6130 showed no anti-BVDV activity. Weak antiviral activity was noted against other flaviviruses, including West Nile virus, Dengue type 2, and yellow fever virus. These results indicate that PSI-6130 is a specific inhibitor of HCV. PSI-6130 showed little or no cytotoxicity against various cell types, including human peripheral blood mononuclear and human bone marrow progenitor cells. No mitochondrial toxicity was observed with PSI-6130. The reduced activity against the RdRp S282T mutant suggests that PSI-6130 is an inhibitor of replicon RNA synthesis. Finally, the no-effect dose for mice treated intraperitoneally with PSI-6130 for six consecutive days was > or =100 mg/kg per day.

An electrophysiological insight into visual attention mechanisms underlying schizotypy.

A theoretical framework has been put forward to understand attention deficits in schizophrenia (Luck SJ & Gold JM. Biological Psychiatry. 2008; 64:34-39). We adopted this framework to evaluate any deficits in attentional processes in schizotypy. Sixteen low schizotypal (LoS) and 16 high schizotypal (HiS) individuals performed a novel paradigm combining a match-to-sample task, with inhibition of return (using spatially uninformative cues) and memory-guided efficient visual-search within one trial sequence. Behavioural measures and event-related potentials (ERPs) were recorded. Behaviourally, HiS individuals exhibited a spatial cueing effect while LoS individuals showed the more typical inhibition of return effect. These results suggest HiS individuals have a relative deficit in rule selection - the endogenous control process involved in disengaging attention from the uninformative location cue. ERP results showed that the late-phase of N2pc evoked by the target stimulus had greater peak latency and amplitude in HiS individuals. This suggests a relative deficit in the implementation of selection - the process of focusing attention onto target features that enhances relevant/suppresses irrelevant inputs. This is a different conclusion than when the same theoretical framework has been applied to schizophrenia, which argues little or no deficit in implementation of selection amongst patients. Also, HiS individuals exhibited earlier onset and greater amplitude of the mismatch-triggered negativity component. In summary, our results indicate deficits of both control and implementation of selection in HiS individuals.

Reverse two-hybrid techniques in the yeast Saccharomyces cerevisiae.

Use of the yeast two-hybrid system has provided definition to many previously uncharacterized pathways through the identification and characterization of novel protein-protein interactions. The two-hybrid system uses the bifunctional nature of transcription factors, such as the yeast enhancer Gal4, to allow protein-protein interactions to be monitored through changes in transcription of reporter genes. Once a positive interaction has been identified, either of the interacting proteins can be mutated by site-specific or randomly introduced changes, to produce proteins with a decreased ability to interact. Mutants generated using this strategy are very powerful reagents in tests of the biological significance of the interaction and in defining the residues involved in the interaction. Such techniques are termed reverse two-hybrid methods. We describe a reverse two-hybrid method that generates loss-of-interaction mutations of the catalytic subunit of the Escherichia coli heat-labile toxin (LTA1) with decreased binding to the active (GTP-bound) form of human ARF3, its protein cofactor. While newer methods are emerging for performing interaction screens in mammalian cells, instead of yeast, the use of reverse two-hybrid in yeast remains a robust and powerful means of identifying loss-of-interaction point mutants and compensating changes that remain among the most powerful tools of testing the biological significance of a protein-protein interaction.

Reverse two-hybrid techniques in the yeast Saccharomyces cerevisiae.

Use of the yeast two-hybrid system has provided definition to many previously uncharacterized pathways through the identification and characterization of novel protein-protein interactions. The two-hybrid system uses the bi-functional nature of transcription factors, such as the yeast enhancer Gal4, to allow protein-protein interactions to be monitored through changes in transcription of reporter genes. Once a positive interaction has been identified, either of the interacting proteins can mutate, either by site-specific or randomly introduced changes, to produce proteins with a decreased ability to interact. Mutants generated using this strategy are very powerful reagents in tests of the biological significance of the interaction and in defining the residues involved in the interaction. Such techniques are termed reverse two-hybrid methods. We describe a reverse two-hybrid method that generates loss-of-interaction mutations of the catalytic subunit of the Escherichia coli heat-labile toxin (LTA1) with decreased binding to the active (GTP-bound) form of human ARF3, its protein cofactor.

Event-related potential N270 delayed and enhanced by the conjunction of relevant and irrelevant perceptual mismatch.

Event-related potential studies using delayed match-to-sample tasks have demonstrated the presence of two components, N270 and N400, possibly reflecting the sequential processing of multiple sources of endogenous mismatch. To date, studies have only investigated mismatch between a single cue and target. In this study, we used distractor stimuli to investigate the effect of a secondary source of mismatch distinct from the task-relevant stimulus. Subjects performed two paradigms in which the cue and target could match or mismatch. In one paradigm, task-irrelevant distractors were added--producing a source of task-irrelevant perceptual mismatch. A mismatch-triggered negativity was elicited in both paradigms, but was delayed and enhanced in magnitude in the distractors present paradigm. It is suggested that the distractors may differentially affect mismatch responses through the generation of a task-irrelevant mismatch response.

Quantitative electroencephalography as a biomarker for proneness toward developing psychosis.

The fully dimensional approach to the relationship between schizotypal personality traits and schizophrenia describes schizotypy as a continuum throughout the general population ranging from low schizotypy (LoS) and psychological health to high schizotypy (HiS) and psychosis-proneness. However, no biological markers have yet been discovered that reliably quantify an individual's degree of schizotypy and/or psychosis. This study aimed to evaluate quantitative electroencephalographic (qEEG) measures of power spectra as potential biomarkers of the proneness towards the development of psychosis in schizotypal individuals. The resting-state oscillatory brain dynamics under eyes-closed condition from 16 LoS and 16 HiS individuals were analysed for qEEG measures of background rhythm frequency, relative power in δ, θ, low-α, high-α, low-ß, high-ß and low-γ frequency bands, and the high-temporal cross-correlation of power spectra between low- and high-frequency bands observed by averaging signals from whole-head EEG electrodes. HiS individuals at rest locked the thalamocortical loop in the low-α band at a lower-frequency oscillation and displayed an abnormally high level of neural synchronisation. In addition, the high-α band was found to be positively correlated with both the high-ß and low-γ bands unlike LoS individuals, indicating widespread thalamocortical resonance in HiS individuals. The increase of regional alpha oscillations in HiS individuals suggests abnormal high-level attention, whereas the pattern of correlation between frequency bands resembles the thalamocortical dysrhythmia phenomenon which underlies the symptomatology of a variety of neuropsychiatric disorders including schizophrenia. These qEEG biomarkers may aid clinicians in identifying HiS individuals with a high-risk of developing psychosis.