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Biomedical subjects

G M Clark

Publications and source records attributed to G M Clark.

At least 127 records · Page 7Linked to original sources
PubMed1997

The complex between urokinase plasminogen activator and its type-1 inhibitor in breast cancer extracts quantitated by ELISA.

ELISAs for urokinase plasminogen activator (uPA) and plasminogen activator inhibitor type-1 (PAI-1) have shown that tumor levels of these molecules are prognostic parameters in breast cancer as well as other types of cancer. These ELISAs measure the total amount of the given component, including preforms, active, inactive, and complex-bound forms. However, the amount of the active forms of a component may more closely reflect the ongoing level of proteolytic activity and thereby be particularly related to prognosis. Since the inactive complex between uPA and PAI-1 can only be formed the active forms of the individual components, we have developed a sensitive and specific uPA:PAI-1 complex ELISA consisting of a sandwich format with two monoclonal antibodies (MAbs) against PAI-1 as capture antibodies and three biotinylated MAbs against uPA as detector antibodies. The data were collected as kinetic measurements of bound alkaline phosphatase activity. A standard of uPA:PAI-1 complex could be specifically measured in the assay with a detection limit of 8 pg/ml and a linear relationship between signal and complex concentration up to 4 ng/ml. Neither free uPA nor free PAI-1 were detected by this assay and the addition to the internal standard of free PAI-1 in amounts up to 20 ng/ml or uPA did not reduce the detection of complex by the assay. This ELISA was applied to extracts from 20 individual breast cancers. Each tumor was extracted in two different buffers and the median concentration of uPA:PAI-1 complex in the optimal extraction buffer was 0.8 ng/mg protein, range 0.4-2.8 ng/mg protein. Extraction of the tumor tissue at a low pH prevented de novo formation of complex from free uPA and PAI-1 in the tissue without destabilizing preformed uPA:PAI-1 complex. During incubation of the assay plate at neutral pH further uPA:PAI-1 complex formation from free components in the extracts was blocked by p-nitrophenyl guanidinobenzoate (NPGB). Thus, the present assay selectively quantifies preformed complex in tumor extracts and will enable us, for the first time, to evaluate the potential prognostic value of the uPA:PAI-1 complex in cancer.

Animals↗
PubMed1997

Chronic electrical stimulation of the auditory nerve at high stimulus rates: a physiological and histopathological study.

A major factor associated with recent improvements in the clinical performance of cochlear implant patients has been the development of speech-processing strategies based on high stimulation rates. While these processing strategies show clear clinical advantage, we know little of their long-term safety implications. The present study was designed to evaluate the physiological and histopathological effects of long-term intracochlear electrical stimulation using these high rates. Thirteen normal-hearing adult cats were bilaterally implanted with scala tympani electrode arrays and unilaterally stimulated for periods of up to 2100 h using either two pairs of bipolar or three monopolar stimulating electrodes. Stimuli consisted of short duration (25-50 microseconds/phase) charge-balanced biphasic current pulses presented at 1000 pulses per second (pps) per channel for monopolar stimulation, and 2000 pps/channel for bipolar stimulation. The electrodes were shorted between current pulses to minimize any residual direct current, and the pulse trains were presented using a 50% duty cycle (500 ms on; 500 ms off) in order to simulate speech. Both acoustic (ABR) and electrical (EABR) auditory brainstem responses were recorded periodically during the chronic stimulation program. All cochleas showed an increase in the click-evoked ABR threshold following implant surgery; however, recovery to near-normal levels occurred in approximately half of the stimulated cochleas 1 month post-operatively. The use of frequency-specific stimuli indicated that the most extensive hearing loss generally occurred in the high-frequency basal region of the cochlea (12 and 24 kHz) adjacent to the stimulating electrode. However, thresholds at lower frequencies (2, 4 and 8 kHz), appeared at near-normal levels despite long-term electrode implantation and electrical stimulation. Our longitudinal EABR results showed a statistically significant increase in threshold in nearly 40% of the chronically stimulated electrodes evaluated; however, the gradient of the EABR input/output (I/O) function (evoked potential response amplitude versus stimulus current) generally remained quite stable throughout the chronic stimulation period. Histopathological examination of the cochleas showed no statistically significant difference in ganglion cell densities between cochleas using monopolar and bipolar electrode configurations (P = 0.67), and no evidence of cochlear damage caused by high-rate electrical stimulation when compared with control cochleas. Indeed, there was no statistically significant relationship between spiral ganglion cell density and electrical stimulation (P = 0.459), or between the extent of loss of inner (IHC, P = 0.86) or outer (OHC, P = 0.30) hair cells and electrical stimulation. Spiral ganglion cell loss was, however, influenced by the degree of inflammation (P = 0.016) and electrode insertion trauma. These histopathological findings were consistent with the physiological data. Finally, electrode impedance, measured at completion of the chronic stimulation program, showed close correlation with the degree of tissue response adjacent to the electrode array. These results indicated that chronic intracochlear electrical stimulation, using carefully controlled charge-balanced biphasic current pulses at stimulus rates of up to 2000 pps/channel, does not appear to adversely affect residual auditory nerve elements or the cochlea in general. This study provides an important basis for the safe application of improved speech-processing strategies based on high-rate electrical stimulation.

Action Potentials↗
PubMed1997

Acoustic and electric forward-masking of the auditory nerve compound action potential: evidence for linearity of electro-mechanical transduction.

We investigated electro-mechanical transduction within the cochlea by comparing masking of the auditory nerve compound action potential (CAP) by acoustical and electrical maskers. Forward-masking of the CAP reflects the response to the masker of the cochlear location tuned to the probe. Electrical stimulation was delivered through bipolar stimulating electrodes within the basal turn of the scala tympani. The growth of masking of high-frequency probes which excite cochlear locations close to the stimulating electrodes was similar for both acoustic and electrical maskers, suggesting a linear transduction of electrical energy to mechanical energy. Exposure to intense acoustic stimulation caused an equal loss of sensitivity to acoustic and electrical maskers. Masking of lower-frequency probes by electrical maskers increased rapidly with masker current, suggesting the direct electrical stimulation of neural elements. This masking was reduced by the administration of strychnine suggesting a contribution by the efferents towards masking of these low-frequency probes.

Acoustic Stimulation↗
PubMed1997

Estimating mechanical responses to pulsatile electrical stimulation of the cochlea.

This study estimated the mechanical response of the cochlea to pulsatile electrical stimulation of the scala tympani of the cat. The auditory nerve compound action potential evoked by an acoustic probe was forward-masked by a train of charge-balanced biphasic current pulses. Masking as a function of probe frequency reflected the excitation pattern of the response to the masker and resembled the spectrum of the electrical stimulus. Both pulse rate and pulse width influenced the degree of masking. The vibration of a region of the basilar membrane was estimated by recording the local cochlear microphonic evoked by biphasic pulses. The amplitude of the cochlear microphonic was proportional to the amplitude of the spectral component of the electrical stimulus to which the local cochlear microphonic was tuned. These results are consistent with the generation of a mechanical response to the electrical stimulus.

Acoustic Stimulation↗
PubMed1997

Reduction in excitability of the auditory nerve following electrical stimulation at high stimulus rates. II. Comparison of fixed amplitude with amplitude modulated stimuli.

We have previously shown that acute electrical stimulation of the auditory nerve using charge-balanced biphasic current pulses presented continuously can lead to a prolonged decrement in auditory nerve excitability (Tykocinski et al., Hear. Res. 88 (1995), 124-142). This work also demonstrated a reduction in electrically evoked auditory brainstem response (EABR) amplitude decrement when using an otherwise equivalent pulse train with a 50% duty cycle. In the present study we have extended this work in order to compare the effects of electrical stimulation using both fixed amplitude electrical pulse trains and amplitude modulated (AM) pulse trains that more accurately model the dynamic stimulus paradigms used in cochlear implants. EABRs were recorded from guinea pigs following acute stimulation using AM trains of charge-balanced biphasic current pulses. The extent of stimulus-induced reductions in the EABR were compared with our previous results using either fixed amplitude continuous, or 50% duty cycle pulse trains operating at 0.34 microC/phase (2 mA, 170 micros/phase) at 400 or 1000 pulses/s (Tykocinski et al., Hear. Res. 88 (1995) 124-142). The AM pulse train, operating at the same rates, was based on a 1-s sequence of the most extensively activated electrode of a Nucleus Mini-22 cochlear implant using the SPEAK speech processing strategy exposed to 4-talker babble, and delivered the same total charge as the fixed amplitude 50% duty cycle pulse train. Two hours of continuous stimulation induced a significant, rate-dependent reduction in auditory nerve excitability, and showed only a slight post-stimulus recovery for monitoring periods of up to 6 hours. Following 2 or 4 h of stimulation using an otherwise equivalent pulse train with a 50% duty cycle or the AM pulse train, significantly less reduction in the EABR was observed, and recovery to pre-stimulus levels was generally rapid and complete. These differences in the extent of the recovery between the continuous waveform and both the 50% duty cycle and AM waveforms were statistically significant for both 400 and 1000 pulses/s stimuli. Consistent with our previous results, the stimulus changes observed using AM pulse trains were rate dependent, with higher rate stimuli evoking more extensive stimulus-induced changes. The present findings show that while stimulus-induced reductions in neural excitability are dependent on the extent of stimulus-induced neuronal activity, the use of an AM stimulus paradigm further reduces post-stimulus neural fatigue.

Animals↗
PubMed1997

Histological and physiological effects of the central auditory prosthesis: surface versus penetrating electrodes.

To rehabilitate profoundly deaf patients who are not suitable for cochlear implants, central auditory prostheses have been implanted. To compare two possible electrode configurations - penetrating and surface ones - electrical stimulation of the cochlear nucleus with both types of arrays was tested on guinea pigs and cats. Electrophysiological, autoradiographic and histological measures were used to study effects of the central auditory prostheses on the auditory pathway. The results showed that a successful electrically evoked auditory brainstem response could be recorded with both surface and penetrating electrodes in cats and guinea pigs. In guinea pigs the penetrating electrodes had advantages over surface arrays in the sense of lower thresholds and wider dynamic ranges. In cats penetrating electrodes showed lower thresholds than surface ones. In cats and guinea pigs stimulated with either surface or penetrating electrodes, evoked 2-deoxyglucose (2-DG) label was found in the auditory pathway from the cochlear nucleus to the inferior colliculus. No non-auditory tissues were found with evoked 2-DG label. Histological results showed that in subdivisions of the guinea pig cochlear nucleus stimulated with penetrating electrodes the neurone density was decreased, and the mean soma area was increased compared with the control side. In the cat, penetrating electrodes were associated only with increased mean soma area in parts of the stimulated cochlear nucleus. These results suggest that the physiological advantages of penetrating electrodes over surface ones were achieved with some trade-off in safety, especially in the guinea pig.

Animals↗
PubMed1997

Prognostic significance of PAI-1 and uPA in cytosolic extracts obtained from node-positive breast cancer patients.

Cancer cell invasion is accomplished by the concerted action of several extracellular proteolytic enzyme systems, one of which is the urokinase plasminogen activation system. The different components of this system. e.g. urokinase plasminogen activator (uPA), its receptor uPAR, as well as its main inhibitor plasminogen activator inhibitor type 1 (PAI-1) have all been shown to have prognostic value in breast cancer, i.e. high tumor levels are associated with a poor prognosis. In order to further substantiate the prognostic value of uPA and PAI-1, we have tested the cutpoints (median values and optimized outpoints) from our first study (Cancer Res 53: 2513-2521, 1993) in an independent group of breast cancer patients. Breast cancer cytosols from 100 premenopausal and 150 post-menopausal node positive patients were included. The median observation time was 80 months (range 49-145). Univariate analysis showed that high PAI-1 levels (above the median PAI-1 value) were significantly associated with short recurrence-free survival (RR: 1.65; 95% CI: 1.04-2.63; P = 0.03) and short overall survival (RR: 2.46; 95% CI: 1.52-3.96; P = 0.0001) in postmenopausal patients. Postmenopausal patients with high uPA levels (above the median uPA value) had a significantly shorter recurrence-free survival (RR: 2.04; 95% CI: 1.17-3.56; P = 0.01) and overall survival (RR: 2.07; 95% CI: 1.16-3.70; P = 0.01) than patients with low uPA values. Nearly identical results were obtained when using the optimized PAI-1 or uPA value. In a Cox multivariate analysis which included other established prognostic factors, high PAI-1 was found to be an independent prognostic variable predicting short overall survival with a relative risk of 2.27 in postmenopausal women, and high uPA was found to be an independent prognostic variable predicting short recurrence-free survival with a relative risk of 1.86 in postmenopausal women. The present study indicates that uPA and PAI-1 are independent and significant prognostic variables in subsets of breast cancer patients.

Analysis of Variance↗
PubMed1997

The effect of loudness imbalance between electrodes in cochlear implant users.

OBJECTIVE: The aim was to determine the effect of loudness imbalance between electrodes in patients using the 22-electrode cochlear implant (Cochlear Pty Ltd). It was hypothesized that speech perception scores would be greater when the loudness of electrodes was balanced at the comfort (C) levels than when the C levels were unbalanced. DESIGN: Ten adult patients received a monosyllabic word test (CNC) words) in quiet and a sentence test (CUNY sentences) in noise under two conditions: with C levels balanced for equal loudness and with unbalanced C levels. RESULTS: When the C levels across electrodes were pseudo-randomly unbalanced by 0 to +/- 20% of the electrodes' dynamic ranges (20% unbalancing), 6 of the 10 subjects showed a significant drop in sentence perception scores. Of these patients, none had a significant decrease in perception when the degree of unbalancing was halved. Of the four patients who showed no change with 20% unbalancing, three revealed a significant decline in sentence perception when the degree of unbalancing was doubled. There also were significant group effects for phonemes on the word test as well as for sentences in noise for the 20% unbalancing. CONCLUSIONS: The implications for clinical practice are that it is important to balance the C levels and that clinicians should be encouraged to refine methods for setting C levels in very young children, who may be using unbalanced MAPs. Nevertheless, although most patients revealed a statistically significant drop in sentence perception with 20% imbalance of the C levels, the changes in percentage scores often were only small.

Adolescent↗
PubMed1997

Changes in synthetic and natural vowel perception after specific training for congenitally deafened patients using a multichannel cochlear implant.

OBJECTIVE: The aim was to determine whether the ability to use place-coded vowel formant information could be improved after training in a group of congenitally deafened patients, who showed limited speech perception ability after cochlear implant use ranging from 1 yr 8 mo to 6 yr 11 mo. A further aim was to investigate the relationship between electrode position difference limens and vowel recognition. DESIGN: Three children, one adolescent, and one young adult were assessed with synthesized versions of the words/hid, head, had, hud, hod, hood/containing three formants and with a natural version of these words as well as with a 12-alternative, closed-set task containing monosyllabic words. The change in performance during a nontraining period was compared to the change in performance after 10 training sessions. RESULTS: After training, two children showed significant gains on a number of tests and improvements were consistent with their electrode discrimination ability. Difference limens ranged from one to three electrodes for these patients as well as for two other patients who showed minimal to no improvements. The minimal gains shown by the final patient could be partly explained by poorer apical electrode position difference limen. CONCLUSIONS: Significant gains in vowel perception occurred post-training on several assessments for two of the children. This suggests the need for children to continue to have aural rehabilitation for a substantial period after implantation. Minimal improvements, however, occurred for the remaining patients. With the exception of one patient, their poorer performance was not associated with poorer electrode discrimination.

Adolescent↗
PubMed1997

Pitch and loudness estimation for single and multiple pulse per period electric pulse rates by cochlear implant patients.

Numerical estimates of loudness and pitch for electric pulse rates were obtained from 14 patients using the 22 electrode cochlear implant manufactured by Cochlear Limited. Six patients were postlinguistically deafened adults, and eight patients were adults and children who became deaf very early in life. Comparisons were made between two types of pulse rate patterns. The SPP pulse pattern presented a single pulse every period, the inverse of the pulse rate. The MPP pulse pattern presented multiple pulses in the first half of the period, using a rate of 1000 pulses/s, with no stimulation in the second half of the period. The pulse rates used for the SPP and MPP pulse patterns were 71.4-500 pulses/s, which corresponded to periods of 14-2 ms. For the SPP pulse pattern, the total number of pulses over the duration of the stimulus increased with increases in pulse rate, while for the MPP pulse pattern, the total number of pulses remained constant. Pitch and loudness estimates were obtained from the postlinguistically deafened patients for the SPP and MPP pulse patterns, and from the early-deafened patients for the MPP pulse pattern. Loudness estimates for the SPP pulse pattern increased with increases in pulse rate for all postlinguistically deafened patients. Loudness estimates for the MPP pulse pattern decreased with increases in pulse rate for three of the six postlinguistically deafened patients and for six of the eight early-deafened patients. For the other patients (three postlinguistically deafened and two early-deafened), loudness estimates marginally increased with increases in pulse rate. Pitch estimates for the SPP and MPP pulse patterns increased with increases in pulse rate for the six postlinguistically deafened patients. For the early-deafened patients, pitch estimates for the MPP pulse patterns increased with increases in pulse rate for only five of the eight patients. For the other three early-deafened patients, pitch estimates were similar to the loudness estimates and decreased with increases in pulse rate.

Adolescent↗
PubMed1997

Psychophysical studies with two binaural cochlear implant subjects.

Psychophysical studies have been completed with two binaural cochlear implant patients. In our earlier studies [van Hoesel et al., J. Acoust. Soc. Am. 94, 3187-3189 (1993); R. J. M. van Hoesel and G. M. Clark, Ann. Otol. Rhinol. Laryngol. Suppl. 106 104, 233-235 (1995)], lateralization experiments showed good sensitivity to interaural amplitudes but poor sensitivity to interaural time delays when compared with normal hearing subjects. In the studies presented here, both temporal and binaural intensity interactions were further explored. Interaural time delay (ITD) perception was investigated using direct measurement of the just-noticeable difference (jnd) in ITD. Both rate and place of stimulation were varied. Binaural rate discrimination was measured and compared with monaural rate perception. Binaural intensity interaction was explored for matched and unmatched place conditions by means of loudness summation and central masking studies. Results showed that ITDs for interaural time delays were large when compared to normal hearing, even when place of stimulation on each of the two sides was carefully matched. The jnds in ITD were similar for stimulation rates from 50 to 200 pps, and increased at 300 pps. Rate difference limens experiments showed similar results for diotic and monaural stimuli, but improved jnds for dichotic presentation at stimulation rates below 150-200 pps. Binaural intensity interactions showed loudness summation effects with both patients, for matched as well as unmatched place conditions. Central masking was also observed with both subjects, although it was not found to be place dependent.

Cochlear Implants↗
PubMed1997

bcl-2, p53, and response to tamoxifen in estrogen receptor-positive metastatic breast cancer: a Southwest Oncology Group study.

PURPOSE: To test the hypothesis that high bcl-2 expression and accumulation of p53 protein, both of which should inhibit apoptosis, are associated with a poorer tamoxifen response and a more aggressive clinical course in estrogen receptor (ER)-positive metastatic breast cancer. METHODS: A total of 205 paraffin-embedded tumor blocks were evaluated for nuclear p53 (a marker of p53 inactivation) and cytoplasmic bcl-2 by immunohistochemistry (IHC). All patients received tamoxifen as initial therapy for metastatic disease. The study began in 1982 and follow-up duration of the 24 patients last known alive is 8 years. RESULTS: Response to tamoxifen and time to treatment failure (TTF) were not significantly associated with p53 status, although patients with higher p53 had a worse survival (P = .008; median, 36 v 20 months). Higher bcl-2 expression was associated with higher levels of ER (P = .02), better response to tamoxifen (62% v 49%; P = .07), longer TTF (median, 9 v 5 months; P = .002), and better survival (median, 40 months v 25 months; P = .009). In multivariate analyses, including ER, progesterone receptor (PgR), and p53, high bcl-2 remained significantly associated with a longer TTF (P = .007) and survival (P = .07). p53 status was a significant factor for shorter survival (P = .05), but not for TTF (P = .61). CONCLUSION: p53 status, as determined by IHC is not significantly associated with response to tamoxifen, although tumors with altered p53 protein are inherently more aggressive. Contrary to expectation, high bcl-2 identifies a relatively indolent phenotype of ER-positive metastatic breast cancer, in which patients experience a better clinical response to tamoxifen and a longer survival.

Adult↗
PubMed1997

Comparing prospective and retrospective evaluations of temporomandibular disorders after orthognathic surgery.

The authors conducted a study to document agreement between prospective examinations performed by trained clinical trial examiners and retrospective surgical chart reviews in identifying signs and symptoms of temporomandibular disorders, or TMD. Only a small fraction of the signs and symptoms identified by clinical trial examiners were documented in the surgical charts. Studies relying on retrospective chart reviews may seriously underestimate signs and symptoms of TMD.

Adolescent↗
PubMed1997

Acute study on the neuronal excitability of the cochlear nuclei of the guinea pig following electrical stimulation.

To help deaf patients who cannot benefit from the cochlear implant due to interruption of the auditory nerve, a central auditory prosthesis has been developed to directly stimulate the cochlear nucleus in the brainstem. The electrode array lies on the surface of the cochlear nucleus and is designed to stimulate at 250 pulses/sec. To examine the safety of this prosthesis, guinea pig cochlear nuclei were stimulated acutely with bipolar surface electrodes using charge-balanced biphasic current pulses at rates of 250, 500 or 1,000 pulses/s and charge intensities of 1.8, 2.8, 3.5 or 7.1 microC/phase cm(-2). The electrically evoked auditory brainstem response (EABR) was used to monitor neuronal excitability of the cochlear nuclei following this acute electrical stimulation. Respiration rate and body temperature were also monitored during the experiment. The amplitudes and latencies of the EABR waves were measured and compared among the before, during and after stimulation periods. The results showed that respiration rate and body temperature remained within normal limits for the duration of the acute stimulation. During and after electrical stimulation, no change was found in the EABR waveform, dynamic ranges and threshold with up to 6 h direct continuous stimulation of the cochlear nucleus. There was no significant change in the amplitudes and latencies of the EABR waves after stimulation. However, a slight temporary reduction in the amplitude of the EABR waves was observed at 30-60 min during the course of acute stimulation using the highest charge density (7.1 microC/phase cm(-2)). This reduction showed a stronger correlation with the stimulus current, charge/phase and charge density than threshold. The present findings suggest that acute bipolar electrical stimulation with surface electrodes at rates up to 1,000 pulses/s and charge density up to 7.1 microC/phase cm(-2) is safe for neuronal excitability of the cochlear nucleus in guinea pig.

Animals↗