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.

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.

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 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.

Reduced attention-related negative potentials in schizophrenic adults.

Event-related potentials were recorded from outpatient adult schizophrenics receiving maintenance doses of neuroleptics and from normal control subjects during performance of a reaction time task and a complex visual discrimination task, the Span of Apprehension. Difference potentials were computed to isolate endogenous activity associated with the processing demands of the Span task. Schizophrenics produce significantly less early endogenous negative activity than do normal subjects. This processing-related negativity reflects pattern matching activity to an attentional trace during the serial scan of the visual icon. We previously reported an identical reduction in processing-related negativity in childhood-onset schizophrenia, suggesting that this deficit is age independent. Both frontal contingent negative variation and an early frontal P3 were larger in the schizophrenics than in normal subjects, suggesting an inappropriate mobilization of nonspecific attentional resources. A later posterior P3 was significantly smaller in schizophrenics than in normal subjects.

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.

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.

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.

Probing the time-course of the auditory oddball P3 with secondary reaction time.

An elementary neural model of the P3 is proposed in which the P3 is held to manifest a brief, widely-distributed, inhibitory event. A preliminary and indirect test of the model is described using secondary-task methodology. Manual reaction times were measured to probe clicks delivered during the presumed time-course of an auditory oddball P3. We observed that reaction times to probes presented after oddball stimuli were significantly slowed as compared to reaction times to probes presented after standards. The latency of maximum reaction time slowing corresponded generally to the latency of the P3. The latency of maximum reaction time slowing did not respond to a manipulation varying the latency of the P3. Thus, some of the obtained results were consistent with the P3-inhibition hypothesis, whereas others were not. Secondary-task methodology may provide a valuable new approach to understanding the late event-related potentials.

Reduced attention-related negative potentials in schizophrenic children.

ERPs were recorded from normal and schizophrenic children during performance of a reaction time task (RT) followed by a complex visual discrimination, the span of apprehension task (Span), sensitive to vulnerability factors in schizophrenia. Subjects responded rapidly to the onset of the visual arrays in the RT condition and differentially to the presence of 1 of 2 target letters in the Span condition. The EEG was recorded at 19 scalp sites and ERPs included activity 1 sec before through 1 sec after Span array onset. Difference potentials (Span-RT) were computed to remove unvarying exogenous activity, thus isolating endogenous activity associated with the processing demands of the Span task. When RT and Span task ERPs are compared, schizophrenic children produced a significantly smaller than normal increment in endogenous negative activity. This endogenous negativity differed in its topography and time course from the exogenous components (P1, N1 and P2), and most likely reflects attentional effort associated with serial search, pattern recognition and stimulus identification. We believe that the current results support the position that schizophrenics are impaired in their ability to allocate adequate attentional resources for the processing of the Span stimuli. It is important to note that this deficit is apparent quite early in discriminative processing.

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.

rHuEPO treatment improves brain and cognitive function of anemic dialysis patients.

Twenty-four patients with chronic renal failure, stabilized on hemodialysis, were treated with recombinant human erythropoietin. Before treatment, all patients were anemic (mean Hct = 23.7%). Hematocrits reached normal levels (36.5%) after three months of treatment. Brain event-related potentials and neuropsychological tests were used to assess changes in brain and cognitive functions associated with the correction of anemia. Assessments were done prior to and after three and twelve months of rHuEPO treatment. The P3 component of the event-related potential increased in amplitude significantly with treatment, while its latency was unaffected. Of the four neuropsychological tests administered, scores on two improved significantly with treatment, and the other two approached significance. Taken together, these findings suggest that the correction of anemia to hematocrits near normal in uremic patients by rHuEPO treatment improves brain and cognitive function by raising levels of sustained attention, thus increasing speed and efficiency of scanning and perceptual-motor functions and enhancing learning and memory. These findings also suggest that anemia, either directly or indirectly, may impair brain function.

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 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.

Effect of murine recombinant interleukin-1 on brain oedema in the rat.

We investigated the effects of murine recombinant interleukin-1 (rIL-1, Du Pont) in vivo in the normal rat brain and here report both local and systemic effects of centrally administered rIL-1. Normal rats were given single or multiple atraumatic doses of either rIL-1 or and equal volume (5 microliters) of vehicle for control comparison. All dosages of intraparenchymal rIL-1 produced a uniform a hyperthermic response and concomitant lethargy. There was a related anorexia beyond fever duration. Histologic examination of intraparenchymal injection tracts revealed fibrillary whorls of oedema and a cellular infiltrate surrounding the rIL-1 tract, while similar changes were less prominent in control injection tracts. Repeated high doses of rIL-1 produced significantly higher concentrations of brain water as measured by the gravimetric technique. We conclude that rIL-1 is not only a potent chemoattractant, but is also an edigematic agent when administered in high doses.

Recombinant human erythropoietin treatment may improve quality of life and cognitive function in chronic hemodialysis patients.

Medical, psychological, and social adaptation (quality of life) as well as cognitive function were studied in 15 chronic stable hemodialysis patients before the onset of treatment with recombinant human erythropoietin (r-HuEPO), 1 month after stabilization of normal hematocrit levels, and 10 to 15 months after treatment onset. After r-HuEPO treatment, subjects had significantly higher hematocrits, markedly improved energy levels, and marginally improved global health. r-HuEPO treatment was also associated with progressively decreased levels of subject mood disturbance and dialysis-related stresses. Subjects had no increased participation in paid employment and only minimally increased participation in social and leisure activities at posttreatment data points. There was no significant improvement in cognitive function after treatment. r-HuEPO treatment appears to be associated with higher energy levels, significant psychological benefits, and minimal improvements in social adaptation. The effects on cognitive function merit further study.

Longitudinal P300 latency changes in Alzheimer's disease.

A longitudinal study of the changes in latency of the P300 (P3) wave of the auditory event-related brain potential was undertaken in a group of 18 thoroughly screened and diagnosed possible and probable Alzheimer's disease (pAD) patients and 15 normal controls. On initial recording, P3 latency was significantly prolonged in the pAD group by more than 1.5 standard deviations (40 msec) beyond the normal group. Over the course of the next 3 years, the rate of increase in P3 latency was significantly greater for the patient group than for the controls. The rate of change in P3 latency may reflect accelerated senescence in Alzheimer's disease. Development of the auditory P300 as a marker of neurobiological processes in aging and dementia is discussed.

Relationship of dialysis modality and other factors to cognitive function in chronic dialysis patients.

To determine if dialysis modality may be an independent factor in the level of cognitive function in chronic dialysis patients, cognitive function was studied in 17 pairs of continuous ambulatory peritoneal dialysis (CAPD) and center hemodialysis (CHD) subjects matched for sex, age, diabetic status, and interval since dialysis onset. Data on current metabolic, medical, psychological, and vocational function status were obtained. Neuropsychological (NP) measures included the Number Cancellation Protocol (NCP), Trailmaking test forms A and B (TMT A, TMT B), Symbol Digit Modalities (SDM), and the Rey Auditory Verbal Learning Test (RAVLT). The CAPD subject group had consistently more efficient cognitive function than the CHD subject group. Regardless of modality, the groups of subjects under age 51 and those who were vocationally active had significantly better NP performance. No cognitive function differences were found in groups categorized by sex or duration of dialysis. Creatinine levels were more highly correlated with NP scores than were BUN levels, with higher creatinine levels associated with better cognitive function. Serum calcium, CO2, total protein, albumin, and SGOT levels also were correlated with NP scores. CAPD may be more effective than HD in reversing uremic encephalopathy by mechanisms mostly unrelated to serum creatinine and BUN levels. Longitudinal studies will be needed to determine if dialysis modality is an independent factor in the degree of reversal of uremic encephalopathy.

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.

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.