Systematic Review on EEG Analysis to Diagnose and Treat Autism by Evaluating Functional Connectivity and Spectral Power.

An abnormality in neural connectivity is linked to autism spectrum disorder (ASD). There is no way to test the concept of neural connectivity empirically. According to recent network theory and time series analysis findings, electroencephalography (EEG) can assess neural network architecture, a sign of activity in the brain. This systematic review aims to evaluate functional connectivity and spectral power using EEG signals. EEG records the brain activity of an individual by displaying wavy lines that depict brain cells' communication through electrical impulses. EEG can diagnose various brain disorders, including epilepsy and related seizure illness, brain dysfunction, tumors, and damage. We found 21 studies using two of the most common EEG analysis methods: functional connectivity and spectral power. ASD and non-ASD individuals were found to differ significantly in all selected papers. Due to high heterogeneity in the outcomes, generalizations cannot be drawn, and no single method is currently beneficial as a diagnostic tool. For ASD subtype delineation, the lack of research prevented the evaluation of these techniques as diagnostic tools. These findings confirm the presence of abnormalities in the EEG in ASD, but they are insufficient to diagnose. Our study suggests that EEG is useful in diagnosing ASD by evaluating entropy in the brain. Researchers may be able to develop new diagnostic methods for ASD which focuses on particular stimuli and brainwaves if they conduct more extensive studies with higher numbers and more rigorous study designs.

A Digital Filter-Based Method for Diagnosing Speech Comprehension Deficits.

OBJECTIVE: To improve the diagnostic efficiency of current tests for auditory processing disorders (APDs) by creating new test signals using digital filtering methods. METHODS: We conducted a prospective study from August 1, 2014, to August 31, 2019, using 3 low speech redundancy tests with novel test signals that we created with specially designed digital filters: the binaural resynthesis test and the low pass and high pass filtered speech tests. We validated and optimized these new tests, then applied them to healthy individuals across different age groups to examine how age affected performance and to children with APD before and after acoustically controlled auditory training (ACAT) to assess clinical improvement after treatment. RESULTS: We found a progressive increase in performance accuracy with less restrictive filters (P<.001) and with increasing age for all tests (P<.001). Our results suggest that binaural resynthesis and auditory closure mature at similar rates. We also demonstrate that the new tests can be used for the diagnosis of APD and for the monitoring of ACAT effects. Interestingly, we found that patients having the most severe deficits also benefited the most from ACAT (P<.001). CONCLUSION: We introduce a method that substantially improves current diagnostic tools for APD. In addition, we provide information on auditory processing maturation in normal development and validate that our method can detect APD-related deficits and ACAT-induced improvements in auditory processing.

Hand rest and wrist support are effective in preventing fatigue during prolonged typing.

STUDY DESIGN: Case series (longitudinal). INTRODUCTION: Only few reports concerning the efficacy of commonly used strategies for preventing upper limb occupational disorders associated with prolonged typing exist. PURPOSE OF THE STUDY: We aimed to investigate whether the duration of typing and the use of 2 strategies (hand rest and wrist support) changes muscle physiological response and therefore the electromyography records. METHODS: We enrolled 25 volunteers, who were unfamiliar with the task and did not have musculoskeletal disorders. The subjects underwent 3 prolonged typing protocols to investigate the efficacy of the 2 adopted strategies in reducing the trapezius, biceps brachii, and extensor digitorum communis fatigue. RESULTS: Typing for 1 hour induced muscular fatigue (60%-67% of the subjects). The extensor digitorum communis muscle exhibited the highest percentage of fatigue (72%-84%) after 1 and 4 hours of typing (1 hour, P = .04; 4 hours, P = .02). Fatigue levels in this muscle were significantly reduced (24%) with the use of pause typing (4 hours, P = .045), whereas biceps brachii muscle fatigue was reduced (32%) only with the use of wrist supports (P = .02, after 4 hours). Trapezius muscle fatigue was unaffected by the tested occupational strategies (1 hour, P = .62; 4 hours, P = .85). DISCUSSION: Despite presenting an overall tendency for fatigue detected during the application of the protocols, the assessed muscles exhibited different behavior patterns, depending on both the preventive strategy applied and the muscle mechanical role during the task. CONCLUSION: Hand rest and wrist support can successfully reduce muscle fatigue in specific upper limb muscles during prolonged typing, leading to a muscle-selective reduction in the occurrence of fatigue and thus provide direct evidence that they may prevent work-related musculoskeletal disorders. LEVEL OF EVIDENCE: N/A.

Voice disorder in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a chronic disease characterized by progressive tissue damage. In recent decades, novel treatments have greatly extended the life span of SLE patients. This creates a high demand for identifying the overarching symptoms associated with SLE and developing therapies that improve their life quality under chronic care. We hypothesized that SLE patients would present dysphonic symptoms. Given that voice disorders can reduce life quality, identifying a potential SLE-related dysphonia could be relevant for the appraisal and management of this disease. We measured objective vocal parameters and perceived vocal quality with the GRBAS (Grade, Roughness, Breathiness, Asthenia, Strain) scale in SLE patients and compared them to matched healthy controls. SLE patients also filled a questionnaire reporting perceived vocal deficits. SLE patients had significantly lower vocal intensity and harmonics to noise ratio, as well as increased jitter and shimmer. All subjective parameters of the GRBAS scale were significantly abnormal in SLE patients. Additionally, the vast majority of SLE patients (29/36) reported at least one perceived vocal deficit, with the most prevalent deficits being vocal fatigue (19/36) and hoarseness (17/36). Self-reported voice deficits were highly correlated with altered GRBAS scores. Additionally, tissue damage scores in different organ systems correlated with dysphonic symptoms, suggesting that some features of SLE-related dysphonia are due to tissue damage. Our results show that a large fraction of SLE patients suffers from perceivable dysphonia and may benefit from voice therapy in order to improve quality of life.

The spatial properties of L- and M-cone inputs to electroretinograms that reflect different types of post-receptoral processing.

We studied the spatial arrangement of L- and M-cone driven electroretinograms (ERGs) reflecting the activity of magno- and parvocellular pathways. L- and M-cone isolating sine wave stimuli were created with a four primary LED stimulator using triple silent substitution paradigms. Temporal frequencies were 8 and 12 Hz, to reflect cone opponent activity, and 30, 36 and 48 Hz to reflect luminance activity. The responses were measured for full-field stimuli and for different circular and annular stimuli. The ERG data confirm the presence of two different mechanisms at intermediate and high temporal frequencies. The responses measured at high temporal frequencies strongly depended upon spatial stimulus configuration. In the full-field conditions, the L-cone driven responses were substantially larger than the full-field M-cone driven responses and also than the L-cone driven responses with smaller stimuli. The M-cone driven responses at full-field and with 70° diameter stimuli displayed similar amplitudes. The L- and M-cone driven responses measured at 8 and 12 Hz were of similar amplitude and approximately in counter-phase. The amplitudes were constant for most stimulus configurations. The results indicate that, when the ERG reflects luminance activity, it is positively correlated with stimulus size. Beyond 35° retinal eccentricity, the retina mainly contains L-cones. Small stimuli are sufficient to obtain maximal ERGs at low temporal frequencies where the ERGs are also sensitive to cone-opponent processing.

Alouatta trichromatic color vision: cone spectra and physiological responses studied with microspectrophotometry and single unit retinal electrophysiology.

The howler monkeys (Alouatta sp.) are the only New World primates to exhibit routine trichromacy. Both males and females have three cone photopigments. However, in contrast to Old World monkeys, Alouatta has a locus control region upstream of each opsin gene on the X-chromosome and this might influence the retinal organization underlying its color vision. Post-mortem microspectrophotometry (MSP) was performed on the retinae of two male Alouatta to obtain rod and cone spectral sensitivities. The MSP data were consistent with only a single opsin being expressed in each cone and electrophysiological data were consistent with this primate expressing full trichromacy. To study the physiological organization of the retina underlying Alouatta trichromacy, we recorded from retinal ganglion cells of the same animals used for MSP measurements with a variety of achromatic and chromatic stimulus protocols. We found MC cells and PC cells in the Alouatta retina with similar properties to those previously found in the retina of other trichromatic primates. MC cells showed strong phasic responses to luminance changes and little response to chromatic pulses. PC cells showed strong tonic response to chromatic changes and small tonic response to luminance changes. Responses to other stimulus protocols (flicker photometry; changing the relative phase of red and green modulated lights; temporal modulation transfer functions) were also similar to those recorded in other trichromatic primates. MC cells also showed a pronounced frequency double response to chromatic modulation, and with luminance modulation response saturation accompanied by a phase advance between 10-20 Hz, characteristic of a contrast gain mechanism. This indicates a very similar retinal organization to Old-World monkeys. Cone-specific opsin expression in the presence of a locus control region for each opsin may call into question the hypothesis that this region exclusively controls opsin expression.

Voice disorder in cystic fibrosis patients.

Cystic fibrosis is a common autosomal recessive disorder with drastic respiratory symptoms, including shortness of breath and chronic cough. While most of cystic fibrosis treatment is dedicated to mitigating the effects of respiratory dysfunction, the potential effects of this disease on vocal parameters have not been systematically studied. We hypothesized that cystic fibrosis patients, given their characteristic respiratory disorders, would also present dysphonic symptoms. Given that voice disorders can severely impair quality of life, the identification of a potential cystic fibrosis-related dysphonia could be of great value for the clinical evaluation and treatment of this disease. We tested our hypothesis by measuring vocal parameters, using both objective physical measures and the GRBAS subjective evaluation method, in male and female cystic fibrosis patients undergoing conventional treatment and compared them to age and sex matched controls. We found that cystic fibrosis patients had a significantly lower vocal intensity and harmonic to noise ratio, as well as increased levels of jitter and shimmer. In addition, cystic fibrosis patients also showed higher scores of roughness, breathiness and asthenia, as well as a significantly altered general grade of dysphonia. When we segregated the results according to sex, we observed that, as a group, only female cystic fibrosis patients had significantly lower values of harmonic to noise ratio and an abnormal general grade of dysphonia in relation to matched controls, suggesting that cystic fibrosis exerts a more pronounced effect on vocal parameters of women in relation to men. Overall, the dysphonic characteristics of CF patients can be explained by dysfunctions in vocal fold movement and partial upper airway obstruction, potentially caused by the accumulation of mucus and chronic cough characteristic of CF symptomatology. Our results show that CF patients exhibit significant dysphonia and suggest they may potentially benefit from voice therapy as a parallel treatment strategy.

A new low cost wide-field illumination method for photooxidation of intracellular fluorescent markers.

Analyzing cell morphology is crucial in the fields of cell biology and neuroscience. One of the main methods for evaluating cell morphology is by using intracellular fluorescent markers, including various commercially available dyes and genetically encoded fluorescent proteins. These markers can be used as free radical sources in photooxidation reactions, which in the presence of diaminobenzidine (DAB) forms an opaque and electron-dense precipitate that remains localized within the cellular and organelle membranes. This method confers many methodological advantages for the investigator, including absence of photo-bleaching, high visual contrast and the possibility of correlating optical imaging with electron microscopy. However, current photooxidation techniques require the continuous use of fluorescent or confocal microscopes, which wastes valuable mercury lamp lifetime and limits the conversion process to a few cells at a time. We developed a low cost optical apparatus for performing photooxidation reactions and propose a new procedure that solves these methodological restrictions. Our "photooxidizer" consists of a high power light emitting diode (LED) associated with a custom aluminum and acrylic case and a microchip-controlled current source. We demonstrate the efficacy of our method by converting intracellular DiI in samples of developing rat neocortex and post-mortem human retina. DiI crystals were inserted in the tissue and allowed to diffuse for 20 days. The samples were then processed with the new photooxidation technique and analyzed under optical microscopy. The results show that our protocols can unveil the fine morphology of neurons in detail. Cellular structures such as axons, dendrites and spine-like appendages were well defined. In addition to its low cost, simplicity and reliability, our method precludes the use of microscope lamps for photooxidation and allows the processing of many labeled cells simultaneously in relatively large tissue samples with high efficacy.

Flicker ERGs representing chromaticity and luminance signals.

PURPOSE: To test the hypothesis that electroretinograms (ERGs) reflect luminance activity when measured at high temporal frequencies and chromatic activity when measured near 12 Hz. METHODS: The authors measured the responses to stimuli in which the output of red and green light-emitting diodes was modulated in counterphase at different ratios, varying the luminance content in the stimulus while keeping the red-green chromatic contrast and its phase constant. RESULTS: The high temporal frequency electroretinography was determined mainly by the luminance contrast. At 12 Hz, electroretinographic response amplitudes and phases primarily reflected the red-green chromatic content of the stimulus. Control experiments, performed with a deuteranopic subject and with stimuli that silenced the rods and S-cones, excluded an explanation based on intrusion from rod- and S-cone-driven responses. CONCLUSIONS: It now is possible to perform noninvasive measurements of basic electrophysiological properties of the luminance and chromatic pathways on a retinal level, and their disease-related changes, in human observers.

Electrophysiological effects of guanosine and MK-801 in a quinolinic acid-induced seizure model.

Quinolinic acid (QA) is an N-methyl-D-aspartate receptor agonist that also promotes glutamate release and inhibits glutamate uptake by astrocytes. QA is used in experimental models of seizures studying the effects of overstimulation of the glutamatergic system. The guanine-based purines (GBPs), including the nucleoside guanosine, have been shown to modulate the glutamatergic system when administered extracellularly. GBPs were shown to inhibit the binding of glutamate and analogs, to be neuroprotective under excitotoxic conditions, as well as anticonvulsant against seizures induced by glutamatergic agents, including QA-induced seizure. In this work, we studied the electrophysiological effects of guanosine against QA-induced epileptiform activity in rats at the macroscopic cortical level, as inferred by electroencephalogram (EEG) signals recorded at the epidural surface. We found that QA disrupts a prominent basal theta (4-10 Hz) activity during peri-ictal periods and also promotes a relative increase in gamma (20-50 Hz) oscillations. Guanosine, when successfully preventing seizures, counteracted both these spectral changes. MK-801, an NMDA-antagonist used as positive control, was also able counteract the decrease in theta power; however, we observed an increase in the power of gamma oscillations in rats concurrently treated with MK-801 and QA. Given the distinct spectral signatures, these results suggest that guanosine and MK-801 prevent QA-induced seizures by different network mechanisms.

[Anthelmintic activity of aqueous and ethanolic extracts of Morinda citrifolia fruit on Ascaridia galli]. / Atividade anti-helmintica dos extratos aquoso e etanolico do fruto da morinda citrifolia sobre Ascaridia galli.

The anthelmintic activity of aqueous and ethanolic extracts of Morinda citrifolia fruit (noni) was evaluated in chicken naturally infected by Ascaridia galli. The anthelmintic activity in vitro was determined in adult parasites. The aqueous and ethanolic extracts were used in the following concentrations: 1.69; 3.37; 6.74; 13.48 e 26.96 mg.mL(-1) and 4.17; 8.34; 16.68; 33.36 and 66.72 mg.mL(-1), respectively. The anthelmintic activity in vivo was determined by the administration of 10 mL.kg(-1) of the aqueous (50.1 mg.mL(-1)) and ethanolic (24.6 mg.mL(-1)) extracts during three consecutive days. Later the chickens were euthanized and necropsy was performed in order to count the remaining helminths. The data were analyzed by the Student-Newman-Keuls test. In the concentrations of 13.48 and 26.96 mg.mL(-1), the aqueous extract demonstrated mortality of 46.67 and 50%, respectively, there was a significative difference from the negative control (P < 0.05). The ethanolic extract presented statistical difference from the negative control (diluent) (P < 0.05) for the concentrations of 33.36 and 66.72 mg.mL(-1), expressed by a mortality rate of 66.67 and 76.67%, respectively. In the in vivo test, the aqueous extract of noni fruit showed 27.08% of elimination, deferring statistically from the control group. There was no statistical difference between the ethanolic extract treatments and the control (P > 0.05). It follows that the anthelmintic activity of noni fruit test showed satisfactory results in vitro, there is a need for studies in higher concentrations in the in vivo test.

Dengue maculopathy: visual electrophysiology and optical coherence tomography.

The objective of this study was to evaluate the visual loss due to dengue fever using retinal and cortical electrophysiology and retinal imaging. The participants were three female patients with low visual acuity after dengue fever. They were evaluated by routine ophthalmological investigations, transient pattern electroretinogram (tPERG), transient pattern visual evoked cortical potential (tPVECP), and retinal optical coherence tomography (retinal OCT). tPERG and tPVECP amplitude (microV) and implicit time (ms) were the parameters evaluated using OCT retinal thickness (microm) and reflectivity. All patients presented low visual acuity and scotomata with or without changes in the oculus fundus. tPERG from two patients showed decreased amplitude or absence of the main components; it was not possible to record a reliable response in the third patient due to excessive blinking. tPVECP at 0.5 cpd was normal in all three patients, while at 2 cpd the main components were absent in one patient and normal in the other two patients. OCT image was abnormal in two patients, one of them with high reflectance areas and another with decreased retinal thickness (the third patient was not studied with this technique).The dengue fever can lead to visual impairment detectable by ophthalmological exams such as angiography, retinography, and OCT imaging, as well as retinal and cortical electrophysiology. Dengue maculopathy which could be caused by vascular alterations and/or aberrant immune response after infection may result in temporary or permanent visual losses.

Developmental sources of conservation and variation in the evolution of the primate eye.

Conserved developmental programs, such as the order of neurogenesis in the mammalian eye, suggest the presence of useful features for evolutionary stability and variability. The owl monkey, Aotus azarae, has developed a fully nocturnal retina in recent evolution. Description and quantification of cell cycle kinetics show that embryonic cytogenesis is extended in Aotus compared with the diurnal New World monkey Cebus apella. Combined with the conserved mammalian pattern of retinal cell specification, this single change in retinal progenitor cell proliferation can produce the multiple alterations of the nocturnal retina, including coordinated reduction in cone and ganglion cell numbers, increase in rod and rod bipolar numbers, and potentially loss of the fovea.

Color discrimination ellipses of trichromats measured with transient and steady state visual evoked potentials.

The purpose of this work is to investigate the use of different forms of visual evoked potentials (VEPs) to measure color discrimination thresholds and to plot color discrimination ellipses (MacAdam, 1942). Five normal trichromats (24.5 +/- 2.6 years-old) were monocularly tested. Stimuli consisted of sinusoidal isoluminant chromatic gratings made from chromaticity pairs located along four different color directions radiating from one reference point of the CIE 1976 chromaticity diagram (u' = 0.225; v' = 0.415). Heterochromatic flicker photometry (HFP) was used to obtain the isoluminance condition for every subject and for all chromaticity pairs. VEPs were elicited using two cycles per degree grating stimuli at three different temporal configurations: transient, onset (300 ms)/offset (700 ms), 1 Hz fundamental frequency; steady-state, onset (50 ms)/offset (50 ms), 10 Hz fundamental frequency; and steady-state pattern reversal at 5 Hz fundamental frequency (10 Hz phase reversal). VEP amplitude was measured using transient VEP N1-P1 components and steady state VEP first (10 Hz) and second (20 Hz) harmonics. VEP amplitude was plotted as a function of chromatic distance in the CIE 1976 color space and the data points were extrapolated to zero amplitude level to obtain chromatic discrimination thresholds. The results were compared with psychophysical measurements performed using the same stimulus configurations and with the pseudoisochromatic method of Mollon-Reffin (one-way ANOVA). For all subjects and all stimulation methods, the ellipses showed small sizes, low ellipticities, and were vertically oriented. Despite some consistent differences in the results obtained with different procedures, there was no statistical difference between ellipses obtained electrophysiologically and psychophysically. For steady state VEPs, ellipses obtained from second harmonic amplitudes were larger and more elongated in the tritan direction than those obtained with first harmonic amplitudes.

Amplitude of the transient visual evoked potential (tVEP) as a function of achromatic and chromatic contrast: contribution of different visual pathways.

We investigated how the stimulation mode influences transient visual evoked potentials (tVEP) amplitude as a function of contrast of achromatic and isoluminant chromatic gratings. The chromatic stimulation probed only responses to the red-green axis. Visual stimuli were monocularly presented in a 5 degrees diameter circle, achromatic and chromatic horizontal gratings, 1 Hz pattern reversal stimulation, and achromatic and chromatic gratings, 300 ms onset per 700 ms offset stimulation. For the achromatic pattern reversal stimulation, a double slope function describes how the P100 amplitude varied as a function of log contrast which had a limb at low-to-medium contrasts and another limb at high contrasts. For the achromatic onset/offset stimulation, C2 amplitude saturated at the highest contrast tested and a single straight line described how it changed along most of the contrast range. Both presentation modes for chromatic gratings resulted in amplitude versus log contrast relations which were well described by single straight lines along most of the contrast range. The results may be interpreted as if at 2 cpd, achromatic pattern reversal stimulation evoked the activity of at least two visual pathways with high and low contrast sensitivity, respectively, while achromatic onset/offset stimulation favored the activity of a pathway with high contrast sensitivity. The neural activity in the M pathway is the best candidate to be the high contrast mechanism detected with pattern reversal and pattern onset/offset VEPs. The activity of color opponent pathways such as the P and K pathways either combined or in isolation seems to be responsible for VEPs obtained with isoluminant chromatic gratings at both presentation modes. When the amplitudes of chromatic VEPs were plotted in the same contrast scale as used for achromatic VEPs, chromatic contrast thresholds had similar values to those of the achromatic mechanism with high contrast sensitivity.

Mercury toxicity in Amazon gold miners: visual dysfunction assessed by retinal and cortical electrophysiology.

Amazonian gold mining activity results in human exposure to mercury vapor. We evaluated the visual system of two Amazonian gold miners (29 and 37 years old) by recording the transient pattern electroretinogram (tPERG) and transient pattern visual evoked potential (tPVEP). We compared these results with those obtained from a regional group of control subjects. For both tPERG and tPVEP, checkerboards with 0.5 or 2 cycles per degree (cpd) of spatial frequency were presented in a 16 degrees squared area, 100% Michelson contrast, 50cd/m2 mean luminance, and 1 Hz square-wave pattern-reversal presentation. Two averaged waveforms (n=240 sweeps, 1s each) were monocularly obtained for each subject in each condition. Both eyes were monocularly tested only in gold miners. Normative data were calculated using a final pooled waveform with 480 sweeps. The first gold miner, LCS, had normal tPERG responses. The second one, RNP, showed low tPERG (P50 component) amplitudes at 0.5 cpd for both eyes, outside the normative data, and absence of response at 2 cpd for his right eye. Delayed tPVEP responses (P100 component) were found at 2 cpd for LCS but the implicit times were inside the normative data. Subject RNP also showed delayed tPVEP responses (all components), but only the implicit time obtained with his right eye was outside the normative data at 2 cpd. We conclude that mercury exposure levels found in the Amazon gold miners is high enough to damage the visual system and can be assessed by non-invasive electrophysiological techniques.

Spatial luminance contrast sensitivity measured with transient VEP: comparison with psychophysics and evidence of multiple mechanisms.

PURPOSE: To compare the spatial luminance contrast sensitivity function (CSF) obtained with transient visual evoked potentials (VEPs) with that obtained with psychophysical measurements. METHODS: The stimuli consisted of horizontal luminance gratings. In the VEP experiments, 0.4, 0.8, 2, 4, 8, and 10 cpd of spatial frequency were used, at 1 Hz square-wave contrast-reversal mode. Eight to 10 Michelson contrasts were used at each spatial frequency. Contrast thresholds were estimated from extrapolation of contrast response functions. Psychophysical sensitivities were obtained with spatial gratings of 0.4, 0.8, 1, 2, 4, 6, 8, and 10 cpd and presented at 1 Hz square-wave contrast-reversal or stationary mode (dynamic and static presentation, respectively). CSF tuning was estimated by calculating the ratio between peak sensitivity and the sensitivity at 0.4 cpd. RESULTS: In all subjects tested (n = 6), VEP contrast-response functions showed nonlinearities-namely, amplitude saturation and double-slope amplitude functions that occurred at low and medium-to-high spatial frequencies, respectively. Mean electrophysiological and psychophysical CSFs peaked at 2 cpd. CSF tuning for electrophysiology and dynamic and static psychophysics were, respectively, 1.08, 1.11, and 1.31. Correlation coefficients (r(2)) between electrophysiological CSF and dynamic or static psychophysical CSF were, respectively, 0.81 and 0.45. CONCLUSIONS: Electrophysiological and psychophysical CSFs correlated more positively when temporal presentation was similar. Spatial frequencies higher than 2 cpd showed that at least two visual pathways sum their activities at high contrasts. At low contrast levels, the results suggest that the transient VEP is dominated by the magnocellular (M) pathway.

Response of carp (Cyprinus carpio) horizontal cells to heterochromatic flicker photometry.

The objective of the present work was to determine the interaction of cone inputs in the response of horizontal cells using heterochromatic flicker photometry (HFP). Intracellular electrophysiological recordings were made in horizontal cells of isolated retinae of carp maintained in physiological solution, with the receptor side up. Sharp glass microelectrodes filled with 3 M KCl solution with resistances between 100 and 120 M Omega were used. Stimuli comprised six cycles of two 6-Hz sinusoidal light waves in counterphase adjusted for the same number of quanta: a green light (550 nm) from a monochromator with a Xenon lamp and an LED red light (628 nm). The stimulation program consisted of 10 steps with the 550-nm wave at constant amplitude, while the 628-nm wave varied in increments of 10% up to 100%, followed by another 10 steps with the 628-nm wave at constant amplitude while the 550-nm wave varied in increments of 10% up to 100%. We recorded responses from four different horizontal cell classes: H1 (monophasic, broadband, n = 37), H2 (biphasic, red-green color-opponent, n = 13), and H3 (biphasic, blue-yellow color-opponent, n = 2) cone horizontal cells; and RH (monophasic, broadband, n = 3) rod horizontal cells. H1 and RH horizontal cells showed a similar cancellation point at a heterochromatic mixture consistent with mixed inputs from 630- and 550-nm cones. No cancellation point was found for the H2 cell class. Fish H1 cells add cone inputs and signal "luminance" in light levels appropriate for cone stimulation. The same occurs with RH cells, which also signal "luminance," but in light levels appropriate for rod work. For both cell classes there is an HFP cancellation point occurring at a combination of 628-nm and 550-nm lights in opposing phase that leads to the cancellation of the cell's response. No cancellation was found for H2 and H3 cells, which are the chromatically opponent horizontal cells in lower vertebrates.

Normal and dichromatic color discrimination measured with transient visual evoked potential.

It would be informative to have an electrophysiological method to study, in an objective way, the effects of mercury exposure and other neurotoxics on human color vision performance. The purpose of the present work was to study human color discrimination by measuring chromatic difference thresholds with visual evoked potential (VEP). Six young normal trichromats (24 +/- 1 years old) and one deutan (26 years old) were tested. The stimuli consisted of sinusoidal isoluminant chromatic gratings made from chromaticity pairs located along four different color directions centered on two reference points. Heterochromatic flicker photometry (HFP) protocol was used to obtain the isoluminance condition for every subject and for all chromaticity pairs. Spatial frequency was 2 cycles/deg. Presentation mode comprised onset (300 ms)/offset (700 ms) periods. As previously described, we found a negative deflection in the VEP which was related to the chromatic difference: as chromatic difference increased, amplitude increased and latency decreased. VEP response amplitude was plotted against distance in the CIE 1976 color space between the grating chromaticities and fitted with a regression line. We found color thresholds by extrapolating the fitting to null amplitude values. The thresholds were plotted in the CIE 1976 color space as MacAdam ellipses. In normal trichromats the ellipses had small size, low ellipticity, and were vertically oriented. In the deutan subject, the ellipses had large size, high ellipticity, and were oriented towards the deutan copunctal locus. The VEP thresholds were similar to those obtained using grating stimuli and psychophysical procedures, however smaller than those obtained using pseudoisochromatic stimuli (Mollon-Reffin method). We concluded that transient VEP amplitude as a function of contrast can be reliably used in objective studies of chromatic discrimination performance in normal and altered human subjects.

Horizontal cell morphology in nocturnal and diurnal primates: a comparison between owl-monkey (Aotus) and capuchin monkey (Cebus).

Horizontal cell morphology was studied in the retina of the nocturnal owl-monkey, Aotus, and compared with that of its diurnal, close relative, the capuchin monkey, Cebus . Cells were initially labeled with DiI and the staining was later photoconverted in a stable precipitated using DAB as chromogen. The sizes of cell bodies, dendritic fields, and axon terminals, number of dendritic clusters, intercluster spacing, and intercone spacing were measured at increasing eccentricities. Two distinct morphological classes of horizontal cells were identified, which resembled those of H1 and H3 cells described in diurnal monkeys. A few examples of a third class, possibly corresponding to the H2 cells of diurnal monkeys, were labeled. Both H1 and H3 cells increased in size and had increasing numbers of dendritic clusters with eccentricity. H3 cells were larger and had a larger number of dendritic clusters than H1 cells. Owl-monkey H1 cells had larger dendritic fields than capuchin monkey H1 cells at all quadrants in the central and midperipheral retinal regions, but the difference disappeared in the far periphery. Owl-monkey and capuchin monkey H1 cells had about the same number of dendritic clusters across eccentricity. As owl-monkey H1 cells were larger than capuchin monkey H1 cells, the equal number of clusters in these two primates was due to the fact that they were more spaced in the owl-monkey cells. H1 intercluster distance closely matched intercone spacing for both the owl-monkey and capuchin monkey retinas. On the other hand, H3 intercluster distance was larger than intercone spacing in the retina of both primates. Owl-monkey H1 axon terminals had 2-3 times more knobs than capuchin monkey H1 axon terminals in spite of having about the same size and, consequently, knob density was 2-3 times higher for owl-monkey than capuchin monkey H1 axon terminals across all eccentricities. The differences observed between owl-monkey and capuchin monkey horizontal cells, regarding the morphology of their dendritic trees and axon terminals, may be related to the differences found in the cone-to-rod ratio in the retina of these two primates. They seem to represent retinal specializations to the nocturnal and diurnal life styles of the owl-monkey and capuchin monkey, respectively.