Correlating the alpha rhythm to BOLD using simultaneous EEG/fMRI: inter-subject variability.

Simultaneous recording of electroencephalogram/functional magnetic resonance images (EEG/fMRI) was applied to identify blood oxygenation level-dependent (BOLD) changes associated with spontaneous variations of the alpha rhythm, which is considered the hallmark of the brain resting state. The analysis was focused on inter-subject variability associated with the resting state. Data from 7 normal subjects are presented. Confirming earlier findings, three subjects showed a negative correlation between the BOLD signal and the average power time series within the alpha band (8--12 Hz) in extensive areas of the occipital, parietal and frontal lobes. In small thalamic areas, the BOLD signal was positively correlated with the alpha power. For subjects 3 and 4, who displayed two different states during the data acquisition time, it was shown that the corresponding correlation patterns were different, thus demonstrating the state dependency of the results. In subject 5, the changes in BOLD were observed mainly in the frontal and temporal lobes. Subject 6 only showed positive correlations, thus contradicting the negative BOLD alpha power cortical correlations that were found in most subjects. Results suggest that the resting state varies over subjects and, sometimes, even within one subject. As the resting state plays an important role in many fMRI experiments, the inter-subject variability of this state should be addressed when comparing fMRI results from different subjects.

A whole head MEG study of the amplitude-modulation-following response: phase coherence, group delay and dipole source analysis.

OBJECTIVE: The amplitude-modulation-following response (AMFR) is the frequency component detectable in the electroencephalogram (EEG) or magnetoencephalography (MEG) corresponding to the modulation frequency of an amplitude modulated tone used as a continuous acoustic stimulus. Various properties of the AMFR depend on modulation frequency, suggesting that different generators along the auditory pathway are involved. The present study addresses these issues on the basis of a whole head MEG experiment. METHODS: AM tones with modulators in the 40 Hz and 80 Hz range were presented unilaterally to 10 normal hearing subjects. Biomagnetic responses were recorded with a 151 channel MEG system. The data analysis concentrated on the phase coherence of the responses, group delays and the estimated location of underlying equivalent dipole sources. RESULTS: MEG AMFR is more reliably detected in the 40 Hz than in the 80 Hz range. Both response amplitude and phase coherence indicate clear bilateral activation over the parietal/temporal region. Dipole source analysis confirms that sources are located in or near the auditory cortex. Group delays at 80 Hz are shorter than at 40 Hz. CONCLUSIONS: In both modulation frequency ranges MEG responses are dominated by activity in the auditory cortex, in apparent contrast with EEG data in the literature, pointing to dominant contributions of thalamic sources to the 80 Hz AMFR.

Decreasing fibrogenesis: an immunohistochemical study of paired liver biopsies following lamivudine therapy for chronic hepatitis B.

BACKGROUND: Activation of hepatic stellate cells is the earliest step in fibrogenesis. Alpha-smooth muscle actin (alpha-SMA), expressed by activated hepatic stellate cells, and C-terminal procollagen alpha1(III) propeptide (PIIICP) are early markers of fibrogenesis and should precede fibrosis. AIM: Determine if suppression of hepatitis B virus replication with lamivudine would decrease fibrogenesis as measured by immunohistochemical markers. METHODS: Paired liver biopsies from patients with hepatitis B before and after therapy with lamivudine (n=47) or placebo (n=33) were studied. alpha-SMA and PIIICP were detected in paraffin-embedded tissue by immunohistochemistry and quantified in a blinded manner by video imaging analysis. RESULTS: Liver biopsies from patients treated with lamivudine showed a significant decrease in alpha-SMA expression (1.06+/-0.23 vs. 0.58+/-0.11, pre vs. post, P<0.05). Placebo recipients had increased levels of alpha-SMA (0.82+/-0.14 vs. 1.32+/-0.21, P<0.05). PIIICP was similarly decreased after lamivudine. Among subjects whose Histologic Activity Index fibrosis score was unchanged or worsened, the mean change in alpha-SMA expression was significantly decreased in the lamivudine group compared with placebo. CONCLUSIONS: Lamivudine decreased markers of hepatic stellate cell activation and collagen synthesis. Immunohistochemical techniques are sensitive for assessing fibrogenesis and will be useful in trials of antiviral and antifibrotic agents.

Amplitude of distortion product otoacoustic emissions in the guinea pig in f(1)- and f(2)-sweep paradigms.

The amplitude versus frequency relations of distortion product otoacoustic emissions (DPOAEs) were studied in the guinea pig, using both the f(1)- and the f(2)-sweep paradigms to vary the primary frequency separation. The amplitude of the DPOAEs 2f(1)-f(2), 3f(1)-2f(2), 4f(1)-3f(2), and 2f(2)-f(1), plotted as a function of DP frequency, exhibited a bandpass structure. The separation of the primaries for which the DPOAE level is maximum is referred to as the optimum ratio f(2)/f(1). For the lower sideband DPOAEs (f(dp)

Repeated inhalation exposure to octamethylcyclotetrasiloxane produces hepatomegaly, transient hepatic hyperplasia, and sustained hypertrophy in female Fischer 344 rats in a manner similar to phenobarbital.

Octamethylcyclotetrasiloxane (D4) has been described as a phenobarbital-like inducer of hepatic enzymes. Phenobarbital (PB) and phenobarbital-like chemicals induce transient hepatic and thyroid hyperplasia and sustained hypertrophy in rats and mice. The extent to which these processes are involved with D4-induced hepatomegaly is not known. The present study has evaluated the effects of repeated inhalation exposure to D4 vapors on hepatic and thyroid cell proliferation and hypertrophy with respect to time and exposure concentration. Female Fischer 344 rats were exposed via whole body inhalation to 0 ppm D4, 700 ppm D4 vapors (6 h/day; 5 days/week), or 0.05% PB in drinking water over a 4-week period. Incorporation of 5'-bromo-2-deoxyuridine (BrdU) and the abundance of proliferating cell nuclear antigen were used as indicators of cell proliferation. Designated animals from each treatment group were euthanized on study days 6, 13, and 27. The effect of D4 exposure concentration on hepatic cell proliferation was evaluated at 0, 7, 30, 70, 150, 300, or 700 ppm. Liver-to-body weight ratios in animals exposed to 700 ppm D4 were increased 18, 20, and 22% over controls while PB-treated animals showed increases of 33, 27, and 27% over controls on days 6, 13, and 27 respectively. Hepatic incorporation of BrdU following exposure to D4 was highest on day 6 (labeling index = 15-22%) and was at or below control values by day 27. This pattern of transient hyperplasia was observed in all hepatic lobes examined and was similar to the pattern observed following treatment with PB.

DPOAE group delays versus electrophysiological measures of cochlear delay in normal human ears.

Group delays of 2 f1-f2 distortion product otoacoustic emissions (DPOAEs) were determined using both f1- and f2-sweep paradigms in 24 normal-hearing subjects. These DPOAE group delays were studied in comparison with cochlear delays estimated from derived band VIIIth nerve compound action potentials (CAPs) and auditory brainstem responses (ABRs) in the same subjects. The center frequencies of the derived bands in the electrophysiological experiment were matched with the f2-frequencies in the DPOAE recording to ensure that DPOAEs and derived CAPs and ABRs were generated at the same places along the cochlear partition, thus allowing for a direct comparison. The degree to which DPOAE group delays are larger in the f2- than in the f1-sweep paradigm is consistent with a theoretical analysis of the so-called wave-fixed model. Both DPOAE group delays are highly correlated with CAP- and ABR-derived measures of cochlear delay. The principal result of this study is that "roundtrip" DPOAE group delay in the f1-sweep paradigm is exactly twice as large as the neural estimate of the "forward" cochlear delay. The interpretation of this notion in the context of cochlear wave propagation properties and DPOAE-generating mechanisms is discussed.

Gene delivery of Cu/Zn-superoxide dismutase improves graft function after transplantation of fatty livers in the rat.

Oxygen-derived free radicals play a central role in reperfusion injury after organ transplantation, and fatty livers are particularly susceptible. Endogenous radical scavengers such as superoxide dismutase (SOD) degrade these radicals; however, SOD is destroyed rapidly when given exogenously. Therefore, an adenoviral vector encoding the Cu/Zn-SOD gene (Ad.SOD1) was used here to test the hypothesis that organ injury would be reduced and survival increased in a rat model of transplantation of fatty livers. Donors received chow diet (untreated), high-fat diet, or ethanol-containing high-fat diet. Some of the ethanol-fed donors were infected either with the gene lacZ encoding bacterial beta-galactosidase (Ad.lacZ), or Ad.SOD1. After liver transplantation, SOD activity and protein expression in liver, survival, histopathology, release of transaminases, free radical adducts in bile, and activation of NF-kappaB, IkappaB kinase (IKK), Jun-N-terminal kinase (JNK), and TNFalpha were evaluated. Ad.SOD1 treatment increased survival dramatically, blunted transaminase release, and reduced necrosis and apoptosis significantly. Free radical adducts were increased two-fold in the ethanol group compared with untreated controls. Ad. SOD1 blunted this increase and reduced the activation of NF-kappaB. However, release of TNFalpha was not affected. Ad.SOD1 also blunted JNK activity after transplantation. This study shows that gene therapy with Ad.SOD1 protects marginal livers from failure after transplantation because of decreased oxygen radical production. Genetic modification of fatty livers using viral vectors represents a new approach to protect marginal grafts against primary nonfunction.

Expression of small heat shock protein alphaB-crystallin is induced after hepatic stellate cell activation.

Using the differential PCR display method to select cDNA fragments that are differentially expressed after hepatic stellate cell (HSC) activation, we have isolated from activated HSCs a cDNA that corresponds to rat alphaB-crystallin. Northern blots confirmed expression of alphaB-crystallin in culture-activated HSCs but not in quiescent HSCs. Western blot analysis and immunocytochemical staining confirmed expression of alphaB-crystallin protein in activated but not quiescent HSCs. alphaB-crystallin is induced as early as 6 h after plating HSCs on plastic and continues to be expressed for 14 days in culture. Expression of alphaB-crystallin was also induced in vivo in activated HSCs from experimental cholestatic liver fibrosis. Confocal microscopy demonstrated a cytoplasmic distribution of alphaB-crystallin in a cytoskeletal pattern. Heat shock treatment resulted in an immediate perinuclear redistribution that in time returned to a normal cytoskeletal distribution. The expression pattern of alphaB-crystallin was similar to that of HSP25, another small heat shock protein, but differed from the classic heat shock protein HSP70. Therefore, alphaB-crystallin represents an early marker for HSC activation.

Improving the accuracy of the boundary element method by the use of second-order interpolation functions.

The boundary element method (BEM) is a widely used method to solve biomedical electromagnetic volume conduction problems. The commonly used formulation of this method uses constant interpolation functions for the potential and flat triangular surface elements. Linear interpolation for the potential on a flat triangular mesh turned out to yield a better accuracy. In this paper, we introduce quadratic interpolation functions for the potential and quadratically curved surface elements, resulting from second-order spatial interpolation. Theoretically, this results in an accuracy that is inversely proportional to the third power of element size. The method is tested on a four concentric sphere geometry, representative for electroencephalogram modeling, and compared to previous solutions of this problem in literature. In addition, a cylindrical test configuration is used. We conclude that the use of quadratic interpolation functions for the potential and of quadratically curved surface elements in BEM results in a significant increase in accuracy and in some cases even a reduction of the computation time with the same number of nodes involved in the calculations.

Oxidants from nicotinamide adenine dinucleotide phosphate oxidase are involved in triggering cell proliferation in the liver due to peroxisome proliferators.

It was shown that 4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio acetic acid (Wy-14,643), a potent peroxisome proliferator, caused rapid oxidant-dependent activation of nuclear factor kappaB (NF-kappaB) in Kupffer cells in vivo and activated superoxide production by isolated Kupffer cells. Here, we tested the hypothesis that NADPH oxidase (NADPH OX) is the source of oxidants increased by Wy-14,643. Indeed, both activation of NF-kappaB and increases in cell proliferation due to a single dose of Wy-14,643 (100 mg/kg) were prevented completely when rats were pretreated with diphenyleneiodonium (1 mg/kg), an inhibitor of NADPH OX. p47phox is a critical subunit of NADPH OX; therefore, p47phox knockout mice were used to specifically address the hypothesis of NADPH OX involvement. In livers of wild-type mice, Wy-14,643 activated NF-kappaB, followed by an increase in mRNA for tumor necrosis factor a. Importantly, these changes did not occur in p47phox knockouts. Moreover, when Kupffer cells were treated with Wy-14,643 in vitro, superoxide production was increased in cells from wild-type but not p47phox-null mice. Finally, when mice were fed a Wy-14,643-containing (0.1%) diet for 7 days, the increase in liver weight and cell proliferation caused by Wy-14,643 in wild-type mice was blocked in p47phox-null mice. Combined, these results are consistent with the hypothesis that Wy-14,643 activates NADPH OX, which leads to NF-kappaB-mediated production of mitogens that causes hepatocellular proliferation characteristic of this class of nongenotoxic carcinogens.

Long-term audiometric follow-up of click-evoked auditory brainstem response in hearing-impaired infants.

Conventional pure-tone thresholds were collected as determined at ages between 4 and 8 years from a group of 163 infants, tested by auditory brainstem response (ABR) in the age range between 1 and 3 years old for objective hearing assessment. The subjects suffered from a variety of degrees and types of sensorineural hearing impairment. The prognostic value of the ABR peak V thresholds in response to 0.1 ms clicks with respect to the behavioural thresholds at octave frequencies from 125 to 8,000 Hz obtained later is evaluated. Correlation between ABR and behavioural thresholds is largest in the 1,000- to 8,000-Hz frequency range. Predicted pure-tone audiograms (mean and SD) were determined for each 10-dB class of ABR thresholds. SDs are in the order of 15 to 18 dB in the 500- to 4,000-Hz range and slightly higher at adjacent frequencies (i.e., somewhat larger than in comparable adult studies). Mean pure-tone thresholds in the 1,000- to 8,000-Hz frequency range are up to 20 dB worse than ABR thresholds, which is opposite to findings in normally-hearing subjects. Thus, with an increasing degree of sensorineural hearing impairment, pure-tone thresholds increase at a significantly higher rate than ABR thresholds. The observation is explained in terms of reduced temporal integration in cochlear hearing loss. ABR thresholds worse than 80 dB nHL are demonstrated to have very limited predictive value with respect to the amount of residual hearing, not only in the low- but also in the high-frequency range. The presence of otitis media during ABR testing is shown to make estimation errors increase to more than 25 dB (SD).

Nuclear factor kappaB in proliferation, activation, and apoptosis in rat hepatic stellate cells.

BACKGROUND/AIMS: Activation of the transcription factor NFkappaB has been demonstrated in activated hepatic stellate cells (HSCs). We investigated the role of NFkappaB in proliferation, in activation, and in TNFalpha-induced apoptosis of HSCs. METHODS: NFkappaB activation was inhibited using an adenovirus expressing an IkappaB dominant negative protein (Ad5IkappaB) in both quiescent and activated HSCs. Quiescent HSCs were infected with Ad5IkappaB or an adenovirus expressing beta-galactosidase (Ad5LacZ). The cells were cultured for 7 days. HSCs activation was determined by cell morphology, smooth muscle alpha-actin (alpha-sma) expression, and steady-state mRNA levels of alpha1(I) collagen as assessed by Western blot and RNase protection assay, respectively. Proliferation was determined in culture-activated HSCs by 3H-thymidine incorporation and direct cell counting. Apoptosis was analyzed by infecting quiescent or activated HSCs with Ad5IkappaB or Ad5LacZ, and then treating with TNFalpha. Apoptosis was demonstrated by determining cell number, assessing nuclear morphology, TUNEL assay and caspase 3 activity. RESULTS: After 7 days in culture no differences were noted between the Ad5IkappaB- and the Ad5LacZ-infected cells in the morphology, alpha-sma expression or in alpha1(I) collagen mRNA levels. Ad5IkappaB infection did not modify proliferation in activated HSCs. TNFalpha induced apoptosis only in Ad5IkappaB-infected activated, but not quiescent HSCs. Apoptosis was initially demonstrated 12 h after exposure to TNFalpha. Twenty-four h after the TNFalpha treatment, 60% of the activated HSCs were apoptotic. CONCLUSION: NFkappaB activity is not required for proliferation or activation of HSCs; however, NFkappaB protects activated HSCs against TNFalpha-induced apoptosis.

Group delays of distortion product otoacoustic emissions: relating delays measured with f1- and f2-sweep paradigms.

A theoretical analysis is presented of group delays of distortion product otoacoustic emissions (DPOAEs) measured with the phase-gradient method. The aim of the analysis is to clarify the differences in group delays D1 and D2, obtained using the f1- and the f2-sweep paradigms, respectively, and the dependence of group delays on the order of the DPOAE. Two models are considered, the place-fixed and the wave-fixed models. While in the former model the generation place is assumed to be invariant with both f1- and f2-sweeps, in the latter model the shift of generation place is fully accounted for. By making a simple local approximation of the cochlear scale invariance, a mathematical conversion from phase-place to phase-frequency gradients is incorporated in the wave-fixed model. Under the assumption that the DPOAE (as recorded at the best f2/f1 ratio) is dominated by the contribution from the generation site and not by, e.g., reflection components, the analysis leads to simple expressions for the ratio and difference between D1 and D2. Validation of the models against experimental data indicates that lower sideband DPOAEs (2f1-f2, 3f1-2f2, 4f1-3f2) are most consistent with the wave-fixed model. Upper sideband components (2f2-f1), in contrast, are not properly described by either the place-fixed or the wave-fixed model, independent whether DPOAE generation is assumed to originate at the f2 or at the more basally located f(dp) characteristic place.

Delivery of Cu/Zn-superoxide dismutase genes with a viral vector minimizes liver injury and improves survival after liver transplantation in the rat.

BACKGROUND: Oxygen-derived free radicals play a central role in pathomechanisms of reperfusion injury after organ transplantation. Endogenous radical scavenger systems such as superoxide dismutase (SOD) degrade toxic radicals; however, SOD is degraded rapidly when given exogenously. Therefore, the hypothesis that treatment of the donor liver with an adenoviral vector encoding the Cu/Zn-SOD gene (Ad-SOD1) would lead to permanent gene expression and therefore protect the organ against injury and increase survival in a rat model of liver transplantation was tested. METHODS: Some donors were infected with Ad-SOD1, whereas untreated grafts and livers infected with the indicator gene lacZ encoding bacterial beta-galactosidase (Ad-lacZ) served as controls. After orthotopic liver transplantation, survival, serum transaminases, and histopathology were evaluated. RESULTS: Approximately 80% of hepatocytes expressed beta-galactosidase 72 hr after injection of Ad-lacZ. Moreover, SOD1 gene expression and activity were increased 3- and 10-fold in the Ad-SOD1 group, respectively. After transplantation, 20-25% of rats treated with Ad-lacZ survived. In contrast, all SOD1-treated animals survived. Transaminases measured 8 hr after transplantation in Ad-SOD1 rats were only 40% of those in controls, which increased 40-fold above normal values. Approximately 20% of hepatocytes in untreated and Ad-lacZ-infected organs were necrotic 8 hr after reperfusion, whereas necrosis was nearly undetectable in grafts from rats treated with Ad-SOD1. CONCLUSIONS: This study provides clear evidence for the first time that gene therapy with Ad-SOD1 increases survival and decreases hepatic injury after liver transplantation. Genetic modification of the liver represents a future approach to protect organs against injury where oxygen-derived free radicals are involved.

Inhibition of chronic rejection of aortic allografts by dietary glycine.

BACKGROUND: Chronic rejection is influenced by a variety of risk factors, including histoincompatibility and ischemia. Glycine, a cytoprotective agent, has been shown to protect against ischemia-reperfusion injury in the liver, inactivate hepatic resident macrophages, minimize cyclosporin A-induced nephrotoxicity, and exhibit immunosuppressive properties in vitro. The aim of this study was to investigate whether dietary glycine could reduce development of chronic rejection. METHODS: Lewis recipients of Fisher-344 abdominal aortic allografts received diets that contained either 5% glycine plus 15% casein or 20% casein as control for 10 weeks. Vascular lesions of aortic isografts and allografts were evaluated quantitatively with image analysis and cell counting. RESULTS: No significant vascular changes were observed in isografts (mean medial areas of 3.3 +/- 0.3x0(5) microm2). However, dramatic intimal thickening (neointimal area 2.1+/-0.3) and medial thinning (1.5+/-0.3) were observed in allografts from rats fed control diet. In contrast, glycine significantly reduced the neointima by 45% (1.2+/-0.3) and protected the media (3.5+/-0.2). This led to intima to media area ratios almost twice as large in the control group as in glycine-fed rats (2.2+/-0.4 vs. 1.1+/-0.3, P<0.05). Moreover, infiltrating leukocytes, especially macrophages, were reduced significantly in the adventitia by glycine. In addition, glycine inhibited proliferation and migration of rat aortic smooth muscle cells in culture by 45 and 60%, respectively. CONCLUSION: These results indicate that dietary glycine minimizes histopathological changes of chronic rejection by reducing the immune response and, in part, by minimizing proliferation and migration of smooth muscle cells.

Corn oil rapidly activates nuclear factor-kappaB in hepatic Kupffer cells by oxidant-dependent mechanisms.

N-6 polyunsaturated fatty acids (N-6 PUFAs), major constituents of corn oil and natural ligands for peroxisome proliferator-activated receptors, increase the rate of growth of established tumors. It has been proposed that chemical peroxisome proliferators increase hepatocyte proliferation by mechanisms involving activation of nuclear factor-kappaB (NF-kappaB) and production of low levels of tumor necrosis factor alpha (TNFalpha) by Kupffer cells; however, how N-6 PUFAs are involved in increased cell proliferation in liver is not well understood. Here, the hypothesis that N-6 PUFAs increase production of mitogens by activation of Kupffer cell NF-kappaB was tested. A single dose of corn oil (2 ml/kg, i.g.), but not olive oil or medium-chain triglycerides (saturated fat), caused an approximately 3-fold increase in hepatocyte proliferation. Similarly, when activity of NF-kappaB in whole rat liver or isolated hepatocytes and Kupffer cells was measured at various time intervals for up to 36 h, only corn oil activated NF-kappaB. Corn oil increased NF-kappaB activity approximately 3-fold 1-2 h after treatment exclusively in the Kupffer cell fraction. In contrast, increases were small and only occurred after approximately 8 h in hepatocytes. The activation of NF-kappaB at 2 h and increases in cell proliferation at 24 h due to corn oil were prevented almost completely when rats were pretreated for 4 days with either dietary glycine (5% w/w), an agent that inactivates Kupffer cells, or the NADPH oxidase inhibitor, diphenyleneiodonium (s.c., 1 mg/kg/day). Furthermore, arachidonic acid (100 microM) activated superoxide production approximately 4-fold when added to isolated Kupffer cells in vitro. This phenomenon was not observed with oleic or linoleic acids. Interestingly, a single dose of corn oil increased TNFalpha mRNA nearly 2-fold 8 h after treatment. It is concluded that corn oil rapidly activates NF-kappaB in Kupffer cells via oxidant-dependent mechanisms. This triggers production of low levels of TNFalpha which is mitogenic in liver and promotes growth of hepatocytes.

Gentle organ manipulation during harvest as a key determinant of survival of fatty livers after transplantation in the rat.

Both in situ organ manipulation during harvest and steatosis reduce survival after liver transplantation via mechanisms involving Kupffer cells; thus, their effect on survival was compared here. Moderate steatosis was induced by a single dose of ethanol to Lewis rats, while long-term administration of ethanol yielded severe steatosis in donor animals. After minimal dissection during the first 12 min, livers were either manipulated gently or left alone for 13 min subsequently. Orthotopic liver transplantation was performed after 1 h of cold storage in UW solution. Ethanol increased hepatic lipid content to a level of moderate or severe steatosis that reduced survival after transplantation from 100% to approximately 70% (P < 0.05). However, gentle manipulation decreased survival to approximately 30% (P < 0.05) in livers from normal, saline-treated rats and in livers from rats fed a high-fat control diet. Moreover, after short- or long-term ethanol administration, manipulation of fatty livers decreased survival from 70% to approximately 13% (P < 0.05). Further, manipulation elevated serum transaminases, total bilirubin, and necrosis significantly about 2- to 20-fold in fatty grafts after transplantation. At the end of harvest, trypan blue distribution time and hypoxia assessed from 2-nitroimidazole binding were elevated significantly about two- to fourfold by manipulation of fatty grafts. Gadolinium chloride, a Kupffer cell toxicant, blocked the detrimental effects of manipulation. These data demonstrate for the first time that, while steatosis is detrimental for survival, organ manipulation plays a much greater role than fat in mechanisms of primary nonfunction.

The prognostic value of electrocochleography in severely hearing-impaired infants.

This paper presents a longitudinal evaluation of electrocochleographic assessment in severely hearing-impaired infants. Electrophysiological data were obtained by transtympanic electrocochleography to tone-burst stimuli at octave frequencies of 500 to 8000 Hz at the age of 0-6 years in a group of 126 subjects. The results are compared with auditory thresholds determined at school age in the same children by means of pure-tone audiometry. Cochlear microphonics could be recorded in virtually all ears, although the majority of subjects had hearing losses of 90 dB and more. Compound action potentials (CAPs) showed waveforms varying from normal to a wide range of abnormalities. Audiometric thresholds correlated generally well with the compound action potential (CAP) thresholds obtained in infancy. The error in the predicted audiometric thresholds is between 15 and 20 dB, as compared with 11 dB reported for more moderate hearing losses. It is shown that, in spite of the high stimulus levels used, substantial frequency-specific threshold information is retained. Occasional large discrepancies in thresholds were often associated with markedly abnormal response waveforms. Among the many cases in which no ABR could be elicited, 68 per cent produced detectable electrocochleographic responses in the 1000-4000 Hz range. It is concluded that electrocochleography is a valuable method for the assessment of residual hearing in infants suspected of having a severe hearing impairment.

Group delays of distortion product otoacoustic emissions in the guinea pig.

This paper presents a comprehensive set of experimental data on group delays of distortion product otoacoustic emissions (DPOAEs) in the guinea pig. Group delays of the DPOAEs with frequencies 2f1-f2, 3f1-2f2, 4f1-3f2, and 2f2-f1 were measured with the phase gradient method. Both the f1- and the f2-sweep paradigm were used. Differences between the two sweep paradigms were investigated for the four DPOAEs, as well as the group delay differences between the DPOAEs. Analysis revealed larger group delays with the f2-sweep paradigm, but only for the lower sideband DPOAEs (with fdp < f1,f2). For the lower sideband cubic distortion product 2f1-f2, the f2-sweep delays were a factor of 1.17-1.54 larger than the f1-sweep delays, depending on frequency. The upper sideband DPOAE 2f2-f1 showed no significant difference between f1- and f2-sweep group delays, except for the highest and lowest f2 frequencies. Comparing the group delays of the DPOAEs for each sweep paradigm separately, equal group delays were found for all four DPOAEs measured with the f1-sweep. With the f2-sweep paradigm on the other hand, the group delays of the three lower sideband DPOAEs occurred to be larger than the group delays of the upper sideband DPOAE 2f2-f1. A tentative interpretation of the data in the context of proposed explanatory hypotheses on DPOAE group delays is given.

Kupffer cell oxidant production is central to the mechanism of peroxisome proliferators.

Increased cell proliferation most likely plays a key role in peroxisome proliferator-induced liver cancer. Recently, Kupffer cells were shown to be responsible for Wy-14,643-induced cell proliferation. However, the mechanism by which peroxisome proliferators activate Kupffer cells is unknown. Since gut-derived endotoxin is a known activator of Kupffer cells, the hypothesis that it is involved was evaluated. Increased cell proliferation and peroxisome induction were unaffected by gut sterilization. Moreover, endotoxin was not detectable in portal blood following treatment with Wy-14,643. Therefore, it is concluded that gut-derived endotoxin is not responsible for Kupffer cell activation. To test the hypothesis that Kupffer cells are activated by Wy-14,643 directly, Kupffer cell superoxide production was measured following treatment in vitro. Wy-14,643 increased superoxide production in a dose-dependent manner (0.1 and 50 microM) with half-maximal stimulation at 2.5 microM. Diethylhexylphthalate (DEHP) and ethylhexanol did not increase superoxide production even at doses 50 times higher than Wy-14,643; however, monoethylhexylphthalate (MEHP) activated superoxide production as effectively as Wy-14,643 with half-maximal stimulation at 5 microM. Treatment with Wy-14,643 for 21 days caused a 2-fold increase in Kupffer cell superoxide production while DEHP did not. Pretreatment of Kupffer cells with staurosporine (0.01-10 pM) completely blocked generation of superoxide demonstrating that protein kinase C is required. Moreover, Wy-14,643 increased Kupffer cell protein kinase C activity 3-fold. Pretreatment of Kupffer cells with the amino acid glycine (0.01-3 mM), which blunts calcium signaling, inhibited Wy-14,643-stimulated superoxide production and increased protein kinase C activity completely. These data are consistent with the hypothesis that potent peroxisome proliferators (Wy-14,643 and MEHP) directly activate Kupffer cell production of oxidants via mechanisms involving protein kinase C. Further, peroxisome proliferator treatments that sustain elevated rates of cell proliferation (e.g. Wy-14,643) activate Kupffer cell superoxide production following long-term dietary treatment supporting the hypothesis that Kupffer cell-derived oxidants are involved in peroxisome proliferator-induced neoplasia.