The effect of thermal processing on the behaviour of peanut allergen peptide targets used in multiple reaction monitoring mass spectrometry experiments.

Mass spectrometry-based methods offer an alternative means of determining allergens in foods. Whilst targeted methods are likely to offer the most robust approach for detection and quantification, little is known about how food processing may affect the behaviour of peptide targets. A systematic study has been undertaken to investigate the effects of thermal processing (boiling, roasting, frying) on the behaviour of a suite of peanut peptide targets representing the major clinically-relevant allergens. Initially the effect of thermal processing on protein extractability was investigated and a mass spectrometry-compatible buffer identified comprising 50 mM Tris-HCl, pH 8.8 containing 50 mM dithiothreitol and 0.04% (w/v) acid labile detergent which was able to extract 45-100% of protein from raw, boiled, roasted and fried peanuts using sonication at 60 °C. Eight peptide targets were identified including two peptides from each cupin allergen, Ara h1 and Ara h3 and four peptides from the prolamin superfamily allergens Ara h2, 6 and 7. AQUA peptide standards were synthesised and used to undertake multiple-reaction monitoring experiments, giving assay sensitivities of 0.1-30 amoles of peptide on-column (3 : 1 signal : noise), calculated limits of quantification between 96-1343 amoles of peptide on-column and a linear dynamic range of 4-5 orders of magnitude. Absolute quantification of individual peanut allergens in thermally processed samples showed that peptide targets in the cupin allergens were more prone to processing-induced effects than those from Ara h2, 6 and 7. Targets flanked by arginine residues showed greater thermostability. Identification of processing-stable targets, coupled with more efficient extraction procedures and a wide dynamic range, shows that targeted mass spectrometry methods have great potential as an additional method for quantifying peanut allergens in complex food matrices.

Auditory frequency-following response: neural or artifact?

An electrical response which reproduces the waveform and frequency of the sound stimulus can be recorded from the central neural pathway for audition. Controversy has existed for some years over whether this frequency-following response (FFR) is neural or an artifact such as remote pickup of the cochlear microphonic or cross talk in the recording system. Two experiments resolve this issue by demonstrating that the frequency-following response depends upon functionally intact neural pathways. The frequency-following response, as well as auditory evoked potentials, is abolished by section of the eighth nerve; it is reversibly abolished by cooling of the cochlear nucleus.

Human auditory frequency-following responses to a missing fundamental.

Both a complex tone perceived as a 365-hertz "missing fundamental" and a 365-hertz pure tone evoked 365-hertz far-field frequency-following responses. Narrow-band masking noise centered at 365 hertz attenuated the responses to the pure tone but not to the complex tone. Results support the concept that perception of the missing fundamental is based on periodic neural activity.

Continuous-processing related ERPS in schizophrenic and normal children.

The continuous performance task (CPT) has proven to be sensitive to schizophrenic impairments. Multichannel event-related potential (ERP) data were recorded from schizophrenic and normal children during performance of easy and hard versions of the CPT. Schizophrenics produced fewer hits, more false alarms, and prolonged reaction times. Poor performance in schizophrenics was associated with four ERP abnormalities: (1) Schizophrenics did not exhibit the normal increase in amplitude of an early-onset, processing-related negativity from nontarget to target stimuli, suggesting a failure to appropriately allocate attentional resources to discriminative processing. (2) Although P3 amplitude to targets was not significantly smaller in schizophrenic children, the distribution of P3 amplitude between target and nontarget responses in the easy and hard versions of the CPT was abnormal, suggesting that schizophrenics differed in the strategic allocation of resources in later stages of CPT processing. (3) In all task conditions schizophrenics showed a parietal negative component with a latency of 400 msec seen in younger, but not older normal children, suggestive of maturational lag. (4) ERP data demonstrated absence of right-lateralized P1/N1 amplitude in schizophrenic children. Taken together these data indicate that at several stages of information processing, schizophrenics are deficient in the control and strategic allocation of processing resources.

Event-related potential correlates of impaired attention in schizophrenic children.

Electrophysiological correlates of focused attention were studied in 13 schizophrenic and 19 age- and gender-matched children. Subjects performed a version of the Continuous Performance Task (CPT) in which a target was designated as any digit from 0 through 9 occurring on two successive stimulus presentations. Signal digits were surrounded by distractor digits which varied in position, value, and number. Event-related potentials (ERPs) elicited by each stimulus of a target pair were recorded from midline and homologous parietal, temporal, and occipital electrode placements. Schizophrenic children made significantly more errors of omission and commission than normal children. The amplitude and time course of the intertrial CNV was the same for both groups. There was a circumscribed amplitude asymmetry, left smaller than right, for the P1/N1 and P2 measures which was greater in normal than in schizophrenic children. The P3 component was significantly larger to the second stimulus of the target pair than to the first for both groups, and larger for the normal than the schizophrenic children to both stimuli.

Continuous-processing--related event-related potentials in children with attention deficit hyperactivity disorder.

Visual information processing in children with attention deficit hyperactivity disorder (ADHD) was studied using event-related potentials recorded during two versions of the Continuous Performance Task (CPT). ADHD children made more errors, and had longer reaction times than normal children on both the single- and dual-target CPT. Event-related potential waveforms were normal in the ADHD children with reference to early processing stages, i.e., contingent negative variation, P1-N1 laterality, and processing negativities, suggesting that ADHD children did not differ in their level of preparedness or their ability to mobilize resources for target identification and categorization. With respect to later processing, P3 amplitude was reduced in the ADHD group, whereas P3 latency was longer than normal. ADHD children had a diminished late frontal negative component, suggestive of reduced involvement in postdecisional processing.

Event-related potential correlates of linguistic information processing in schizophrenics.

Event-related potentials (ERPs) were recorded from adult schizophrenics and age- and education-matched normal controls during performance of an idiom recognition task involving judgments of the meaningfulness of idiomatic, literal, and nonsense phrases. Schizophrenics produced more errors and had prolonged reaction times while attempting to correctly differentiate meaningful from meaningless phrases. An ERP correlate of that deficit was a larger than normal N400 to idioms and literals, with no difference in N400 amplitude to nonsense phrases. This result was interpreted as evidence that the influence of the linguistic context provided by the first word of two-word idiomatic and literal phrases is reduced in schizophrenia. Schizophrenics also showed reduced amplitude P300.

Continuous-processing related ERPS in adult schizophrenia: continuity with childhood onset schizophrenia.

BACKGROUND: Previous work with schizophrenic children disclosed deficits on two continuous performance tests (CPTs) and ERP indices of reduced attentional resource allocation. METHODS: The two CPTs were administered to adult schizophrenics and matched control subjects. The simple CPT required only that the subject respond whenever the target digit was displayed. The complex version required a response whenever any digit was displayed on two successive trials. Event-related potentials (ERPs) were recorded during task performance. RESULTS: Schizophrenics had fewer hits on both CPT versions, showed a greater drop in performance from the simple to the complex CPT, and took longer to respond than controls. The processing negativity (Np) showed a greater amplitude increase from nontarget to target in normals than in schizophrenics, and the overlapping P2 component was more negative in normals. P3 latency was longer in schizophrenics, but P3 amplitude did not differ. CONCLUSIONS: Group performance and processing negativity effects replicated those from an earlier study of schizophrenic and normal children administered the same versions of the CPT, suggesting similar abnormalities in the allocation and modulation of information processing resources.

PET and P300 relationships in early Alzheimer's disease.

The P300 (P3) wave of the auditory brain event-related potential was investigated in patients with probable Alzheimer's disease to determine whether P300 latency discriminated these patients from controls and whether prolonged P300 latency correlated with rates of brain glucose metabolism as measured by Positron Emission Tomography. P300 latency was prolonged by more than 1.5 standard deviations from age expectancy in 14 of 18 patients, but none of 17 controls. In these subjects P300 latency was shown to be inversely correlated with relative metabolic rates of parietal and, to a lesser extent, temporal and frontal association areas, but not with subcortical areas.

Event-related potentials in high-functioning adult autistics: linguistic and nonlinguistic visual information processing tasks.

Event-related potentials (ERPs) were recorded from high-functioning adult autistics and age- and IQ-matched normal controls during performance of two non-linguistic information processing tasks, the Continuous Performance Task (CPT) and Span of Apprehension (SPAN), and an Idiom Recognition Task (IRT) involving idiomatic, literal and nonsense phrases. The autistics exhibited behavioral deficits only when attempting to identify idiomatic phrases. The ERP correlate of that deficit was greatly reduced N400 to idioms. In addition, autistics produced larger N1 amplitudes in all tasks, and larger P3s in the IRT and CPT.

Cognitive function, mood and P3 latency: effects of the amelioration of anemia in dialysis patients.

Attention difficulties and psychomotor slowing associated with depressed mood affect the ability of individuals to perform on most neuropsychological tests. It has been suggested that latency of the P3 (P300) component of the event-related EEG potential is an index of neurocognitive status which is not affected by mood. Dialysis patients, who experience diminished dysphoric mood with the reversal of anemia when treated with recombinant human erythropoietin (rHuEPO), were tested for neurocognitive performance, mood and latency of P3. Prior to rHuEPO treatment mood was dysphoric, and neurocognitive testing showed mild deficits, but P3 latency was normal. After treatment, mood improved and neurocognitive test performance was normal. P3 amplitude increased over frontal areas, while P3 latency remained unchanged. Thus, in the case of dysphoric mood, P3 latency may provide a more accurate index of cognitive capacity (as opposed to level of functioning) than neurocognitive test measures.

Information-processing deficits across childhood- and adult-onset schizophrenia.

Event-related potentials were recorded for childhood- and adult-onset schizophrenia subjects performing the span of apprehension (Span) task, which is sensitive to vulnerability factors in schizophrenia. Subjects responded to the onset of the Span arrays in a reaction time condition and then responded differentially to the presence of one of two target letters in the Span condition. While neither the childhood- nor the adult-onset group exhibited abnormalities in preparatory contingent negative variation activity, both groups produced significantly less endogenous negative activity between 100 and 300 ms after Span stimulus onset than age-matched normals. This endogenous negative activity reflects attentional effort associated with serial search and stimulus identification. These results support the position that schizophrenia subjects are impaired in their ability to allocate adequate attentional resources for processing Span stimuli. Moreover, the similarity of this information-processing deficit in the two groups suggests that childhood- and adult-onset schizophrenia lie on a continuum in this regard.

Neural temporal coding of low pitch. I. Human frequency-following responses to complex tones.

The neural basis of low pitch was investigated in the present study by recording a brainstem potential from the scalp of human subjects during presentation of complex tones which evoke a variable sensation of pitch. The potential recorded, the frequency-following response (FFR), reflects the temporal discharge activity of auditory neurons in the upper brainstem pathway. It was used as an index of neural periodicity in order to determine the extent to which the low pitch of complex tones is encoded in the temporal discharge activity of auditory brainstem neurons. A tone composed of harmonics of a common fundamental produces a sensation of pitch equal to that of the 'missing' fundamental. Such signals generate brainstem potentials which are spectrally similar to FFR recorded in response to sinusoidal signals equal in frequency to the missing fundamental. Both types of signals generate FFR which are periodic, with a frequency similar to the perceived pitch of the stimuli. It is shown that the FFR to the missing fundamental is not the result of a distortion product by recording FFR to a complex signal in the presence of low-frequency bandpass noise. Neither is the FFR the result of neural synchronization to the waveform envelope modulation pattern. This was determined by recording FFR to inharmonic and quasi-frequency-modulated signals. It was also determined that the 'existence region' for FFR to the missing fundamental lies below 2 kHz and that the most favorable spectral region for FFR to complex tones is between 0.5 and 1.0 kHz. These results are consistent with the hypothesis that far-field-recorded FFR does reflect neural activity germane to the processing of low pitch and that such pitch-relevant activity is based on the temporal discharge patterns of neurons in the upper auditory brainstem pathway.

Judgments of emotion in words and faces: ERP correlates.

Visual event-related potentials (ERPs) to two types of stimuli (faces and words) were analyzed to determine the effects of the perceived emotional connotations of the stimuli (positive, neutral, or negative) in 10 right-handed normal functioning adult males. Principal component analysis (PCA) of the ERPs reveals 5 factors accounting for over 90% of the ERP waveform variance for both faces and words. In the facial data, two ERP components varied in amplitude according to the perceived emotional connotation of the stimulus. For the P3 component, neutrally rated stimuli produced significantly larger amplitudes than stimuli rated as positive or negative. This effect was lateralized to the left hemisphere. A later positive component, the slow wave (448-616 ms), manifested complementary effects, i.e. faces perceived as positive and negative produced larger amplitudes than those perceived as neutral over the right hemisphere. The verbal stimuli did not result in significant main effects for perceived emotional connotation, but produced subtle connotation-related differences in slow wave topography. Hemispheric asymmetries, unrelated to affective connotation, were evident in the verbal data, manifesting different patterns of lateralization depending on the ERP component. The results suggest that differential processing of emotional connotation affects ERP waveforms and that the effects can be understood in terms of ERP components known to be associated with more general aspects of cognitive processing.

Principal component analysis of ERP differences related to the meaning of an ambiguous word.

Event-related potentials (ERPs) to the noun and verb meanings of/'led/in the single ambiguous phrase 'it was/'led/' were re-analyzed using principal component analysis (PCA). These data had previously been analyzed by SWDA and reported in this journal. PCA defined 3 meaning-related components, comprising 40.3% of the entire data variance. The N150 component was shown to be larger for the noun meaning than for the verb meaning; the P230 epoch differed in its anterior-posterior distribution according to meaning; and N370 for noun responses was relatively more negative at the right posterior lead and positive at the left anterior. All components taken together, the left anterior lead showed the greatest meaning-related difference. Previous analysis by SWDA had resulted in significant discriminant functions for left hemisphere ERPs, but this analysis did not yield a clear definition of the effects of meaning on specific ERP components or of the scalp distributions of meaning-related components. Thus, while the results of both analyses support the interpretation that the perceived meaning of words has a substantial effect on ERP wave forms, PCA appears to provide the clearest definition of the ERP component effects.

Evoked potential waveform differences produced by the perception of different meanings of an ambiguous phrase.

To test the effects of perceived linguistic meaning on evoked potential (EP) waveform, an experiment was performed using the stimulus homophone "led" or "lead" in the single ambiguous phrase "it was /'led/". The phrase was presented aurally in sets of 60 repetitions, instructing the subject before each set as to the specific meaning of the stimulus word to be perceived. Averages of one hundred responses were obtained for each of the two meanings of the stimulus. Responses were recorded from scalp locations over Broca's and Wernicke's areas and homologous right hemisphere loci. Average waveforms evoked by the two meanings of the stimulus word were dissimilar for the left anterior locus and similar for the other three loci as indicated by correlations. The application of the Discrimination Index to these data disclosed that the EP waveform differences observed at the left anterior locus would not be attributed to random variability. Stepwise Discriminant Function Analysis on the same data revealed that responses from individual subjects could be classified according to experimental conditions at a greater-than-chance level. Discrimination was best for responses from left hemisphere loci. There was some commonality of discriminative EP components across subjects, specifically components late in the EP epoch. These data are interpreted as demonstrating an EP correlate of the processing of the contextual meaning of words.

Far-field recorded frequency-following responses: evidence for the locus of brainstem sources.

Two experiments were performed to determine the brainstem origins of the scalp recorded auditory frequency-following response (FFR). The first was a study of FFR onset latency in which responses observed by direct recording from depth electrodes in brainstem auditory nuclei were compared with those obtained from the scalp. The mean onset latency of scalp recorded FFR (5.8 msec) closely approximated the 5.4 msec mean latency of response recorded from within the inferior colliculus (IC). In a second experiment, cooling of the IC greatly reduced or eliminated FFR both within this nucleus and at the scalp. FFR at the medial superior olive was unaffected during cooling. It was concluded that at moderate intensities of stimulation the primary source of scalp recorded FFR is the IC.