Search PubMed⌕ Search

Biomedical subjects

C M Henley

Publications and source records attributed to C M Henley.

34 records · Page 2Linked to original sources

DFMO reduces cortical infarct volume after middle cerebral artery occlusion in the rat.

Ornithine decarboxylase (ODC), a key enzyme in polyamine biosynthesis, is induced in ischemic tissue and may mediate vasogenic edema and delayed neuronal death. We determined the effects of alpha-difluoromethylornithine (DFMO), a specific inhibitor of ODC, on infarct size and ODC activity in a rat model of transient focal ischemia. DFMO blocked the ischemia-induced increase in ODC and significantly reduced infarct volumes by 57-45%, depending upon the treatment regimen. These studies suggest that polyamine metabolism plays a role in the development of cerebral infarction after focal ischemia and that DFMO may be useful in limiting injury after a stroke.

Animals↗

Developmental ototoxicity.

The aminoglycoside antibiotics are active bactericidal agents used for the treatment of tuberculosis and gram-negative bacterial infections. Well over 2 million patients per year receive aminoglycosides; however, a major limitation to their use is ototoxicity (cochleotoxicity and vestibulotoxicity), nephrotoxicity, and neuromuscular blockade. Ototoxicity can result from aminoglycoside administration to any patient population, including adults, fetuses, and preterm and full-term neonates. Preterm infants and experimental animals are hypersensitive to aminoglycoside ototoxicity during the period of anatomic and functional maturation of the inner ear. Developing animals, including humans, are also hypersensitive to loop diuretics and especially to the combination of loop diuretics and aminoglycosides. Ototoxicity depends upon several factors in addition to age, including patient population, dose and duration of therapy, genetic and pharmacokinetic factors, and the particular aminoglycoside used. The molecular mechanisms of developmental hypersensitivity have not yet been determined.

Aminoglycosides↗

Kanamycin inhibits cochlear-renal ODC in neonatal rats.

Ornithine decarboxylase (ODC), a key enzyme in polyamine biosynthesis, is important in development and regeneration. We hypothesize that aminoglycoside inhibition of ODC mediates developmental hypersensitivity to aminoglycoside ototoxicity. Kanamycin effects on ODC activity (decarboxylation of ornithine) in vitro were determined in the postmitochondrial fraction of cochlear and renal homogenates from 11-day-old rats. Kanamycin inhibited cochlear and renal ODC by an uncompetitive mechanism. For the cochlear enzyme, the inhibitor constant (Ki) for kanamycin was 99 +/- 25 mumol/L; for the renal enzyme, the Ki = 1.5 +/- 0.1 mmol/L. In vivo effects of kanamycin on cochlear, renal, brain ODC activity were determined in rats treated with kanamycin (400 mg/kg/day, intramuscularly) or saline during postnatal days 11 through 20, the hypersensitive period for ototoxicity. Rats were killed on postnatal days 12, 14, 16, and 20 and ODC was assayed. Kanamycin significantly inhibited ODC in the lateral wall-organ of Corti and kidney (ANOVA alpha = 0.05), but had no effect on cochlear nerve and no consistent inhibitory effect in the brain. These results suggest that ODC is a potential target of kanamycin in susceptible tissues and may be a contributing factor in developmental sensitivity to the drug by inhibiting repair and developmental processes mediated by ODC.

Animals↗

Postnatal developmental changes in inner ear ornithine decarboxylase (ODC).

Ornithine decarboxylase (ODC), the rate-limiting enzyme in polyamine synthesis, is important in cellular growth, differentiation and development. Although ODC has been quantitated in cochlear tissues of the adult rat, it has not been assessed quantitatively in developing inner-ear tissues. The purpose of the present study was to quantitate ODC in cochlear tissues of the rat during the period of development of hearing. Cochlear ODC was significantly elevated throughout the period of cochlear maturation in that it increased rapidly during the first 10 days, peaked on day 10 and then declined thereafter. ODC in the lateral wall/organ of Corti tissues was significantly higher than in the cochlear nerve in developing, but not in adult rats. Further examination of separate cochlear tissues from 10-day old rats revealed that ODC activity was higher in the organ of Corti than in the lateral wall or cochlear nerve. Postnatal changes in ODC paralleled functional maturation of hearing and the hypersensitive period for aminoglycoside ototoxicity in the rat. Since aminoglycosides have been shown to inhibit ODC in vitro, aminoglycoside inhibition of polyamine synthesis may mediate the hypersensitivity of developing animals to the effects of these drugs.

Age Factors↗

Amino acid assay of vestibular nuclei 10 months after unilateral labyrinthectomy in squirrel monkeys.

Amino acids were assayed by HPLC in bilateral vestibular nuclei from normal squirrel monkeys (n = 3) and those 10-month post-unilateral labyrinthectomy (n = 4). Findings of vestibulo-spinal and vestibulo-oculomotor functions were identical for both groups. No left-right asymmetry of amino acids was found within either group, nor between groups with the exception of GABA: GABA was significantly reduced in the bilateral vestibular nuclei of the 10-month post-lesion animals. This may be indicative of a reduction of cerebello-vestibular inhibitory control which could be secondary to the reduction of excitatory inputs to the system.

Amino Acids↗

Postnatal development of 2f1-f2 otoacoustic emissions in pigmented rat.

Distortion-product emissions at 2f1-f2 were measured in developing, pigmented rats to determine the characteristics of the functional onset and maturation of these emitted responses. Distortion-product testing was conducted on postnatal days 12, 14, 18, 21, 24, 28, and 51. The geometric-mean frequencies of the primaries were tested at one-half octave intervals, between 4 and 11.3 kHz. No emissions were detected on postnatal day 12; however, by day 14, emissions were measured at geometric-mean frequencies between 5.7 and 11.3 kHz, but not at 4 kHz. By the 18th postnatal day, all animals had measurable emissions at 4, 5, 7, 8, and 11.3 kHz. Both the functional onset and maturation of high- to midfrequency emissions developed before those elicited by lower-frequency primaries. Response/growth or input/output functions for the higher frequencies exhibited 'adult-like' properties that included an increase in maximum amplitude and 'saturation' at high levels of stimulation by the third to fourth postnatal week. Maturation of the lower-frequency responses progressed at a slower rate. These results differ somewhat from those reported previously for the neonatal albino rat. However, the present findings were based upon a wider frequency range of primary tones, lower levels of acoustic stimulation, and a measuring system with a significantly lower noise floor. These results are consistent with documented periods of the anatomical maturation of the rat outer hair-cell system and establish a functional 'baseline' for future studies utilizing agents that damage the developing cochlea.

Acoustic Stimulation↗

Effects of cisplatin and thiosulfate upon auditory brainstem responses of guinea pigs.

Two side effects which limit the use of cisplatin in cancer chemotherapy are severe nephrotoxicity and ototoxicity. The concurrent administration of sodium thiosulfate with cisplatin reportedly protects from cisplatin nephrotoxicity, however, protection from ototoxicity has not been documented. The purpose of this study was to examine the efficacy of using thiosulfate to ameliorate the ototoxic effects of cisplatin. Toward this end, the effects of cisplatin alone, cisplatin administered concurrently with sodium thiosulfate (CIS/THIO), and sodium thiosulfate alone on the auditory brainstem response (ABR) of guinea pigs were compared. ABR waveforms, comparing latencies, amplitudes and response thresholds, were monitored before, immediately after, and 30 days post treatment. Sodium thiosulfate administered with cisplatin (CIS/THIO) consistently protected animals from hearing loss and surprisingly yielded significant increases in amplitude when compared to baseline and saline controls. However, ABRs of CIS/THIO animals returned toward baseline values after 30 days.

Animals↗

Inhibition of renal ornithine decarboxylase by aminoglycoside antibiotics in vitro.

The inhibition of renal ornithine decarboxylase (ODC) by aminoglycoside antibiotics was characterized in the postmitochondrial fraction of a kidney homogenate from adult pigmented guinea pigs. Enzymatic activity was defined as the rate of decarboxylation of [14C]ornithine sensitive to a specific ODC inhibitor, alpha-difluoromethylornithine (DFMO). The Km for ornithine was 61 +/- 32 microM. There were two forms of the enzyme with respect to their affinity for pyridoxal phosphate (PLP): (I) Km = 2.1 +/- 1.8 microM; (II) Km = 36.2 +/- 12.7 microM. Putrescine, a known ODC inhibitor, acted competitively on the renal enzyme with Ki = 1.7 +/- 1.4 mM. Aminoglycoside antibiotics inhibited ODC by an uncompetitive mechanism with inhibitor constants of comparable magnitude: neomycin, Ki = 1.3 +/- 0.1 mM; gentamicin, Ki = 1.6 +/- 0.1 mM; kanamycin, Ki = 1.9 +/- 0.2 mM; and netilmicin, Ki = 1.7 +/- 0.2 mM. Neomycin inhibited both forms of the enzyme (low and high affinity for PLP) uncompetitively with similar inhibitor constants (1.5 +/- 0.3 and 1.8 +/- 0.4 mM respectively), suggesting a single mechanism of action. Inhibition of ODC suggests that aminoglycoside-polyamine interactions may be an important component of the sequence of biochemical events associated with aminoglycoside toxicity.

Aminoglycosides↗

Pharmacokinetics of aminoglycoside antibiotics in blood, inner-ear fluids and tissues and their relationship to ototoxicity.

This review critically evaluates the literature on aminoglycoside pharmacokinetics in order to answer the question how fluid and tissue levels of the drugs relate to the development of ototoxic and nephrotoxic side effects. We will summarize the evidence that: (1) aminoglycosides do not accumulate in inner-ear fluids; (2) aminoglycoside levels in fluids do not correlate with the ototoxic potential of a drug, and (3) selective toxicity cannot be explained by selective tissue penetration of the drugs. We suggest that studies of drug disposition at the cellular level after chronic aminoglycoside treatment be conducted to establish whether a cell-specific uptake contributes to the selective toxicity of the aminoglycoside antibiotics. A sequence of biochemical events that may lead to the development of toxicity at the molecular level is briefly described.

Aminoglycosides↗

Inhibition of inner ear ornithine decarboxylase by neomycin in-vitro.

We quantitated the activity of ornithine decarboxylase (ODC) in homogenates and subcellular fractions of inner ear tissues from the rat and guinea pig and demonstrate inhibition of cochlear ODC by the aminoglycoside neomycin. Subcellular fractionation showed the enzyme associated with the post-mitochondrial supernatant fraction in each of the tissues: Specific activities of ODC, defined as alpha-difluoromethylornithine (DFMO)-sensitive decarboxylation of ornithine, in the supernatant fractions of combined inner ear tissues were: guinea pig = 44 +/- 4 pmoles CO2 produced/hour/mg protein, and rat = 133 +/- 30. In the guinea pig, supernatant fractions of the lateral wall tissues (stria vascularis and spiral ligament) had specific activities of 62 +/- 25, those of the organ of Corti (plus VIIIth nerve) 64 +/- 41. The ototoxic aminoglycoside neomycin produced a dose-dependent inhibition of ODC with half-maximal inhibition observed at 50 microM drug and almost complete inhibition at 100 microM. This is the first report of the presence of ODC in the inner ear and its inhibition by neomycin. Since both the ODC-inhibitors, DFMO and neomycin, can cause hearing loss in patients and experimental animals it is suggested that inhibition of ODC may be an important factor in the ototoxicity of these drugs.

Animals↗

Auditory aspects of seizure in the genetically epilepsy prone rat.

The organ of Corti of Genetically Epilepsy Prone Rats was examined anatomically and electrophysiologically using scanning electron microscopy (SEM) and electrophysiological recording of alternating current cochlear potentials (ACCP) and N1, a volume conductor recording of the primary auditory afferent action potentials. ACCPs for GEPRs with low intensity seizures (Acoustic Response Score (ARS) = 2 or 3) and high intensity seizures (ARS = 9) showed similar impairment in cochlear function. Approximately a 25-35 dB shift in input-output functions was present in GEPRs as compared to controls. SEM revealed several types of possible genetic abnormalities which explain the deficits in cochlear function and could serve as the basis for seizure predisposition in these animals.

Acoustic Stimulation↗

Link between functional and morphological changes in the inner ear--functional changes produced by ototoxic agents and their interactions.

Common potentials used to evaluate cochlear function are the ac cochlear potential (ACCP), N1 and the positive dc endocochlear potential (EP). The ACCP is an electrical analogue of the sound stimulus; its source is the electrical activity of the cochlear hair cells. N1 is a volume conductor recorded action potential of the auditory nerve. The EP is the positive polarization of the middle compartment of the cochlea (scala media) with respect to the other compartments (the scalae tympani and vestibuli); the stria vascularis is apparently responsible for the EP. Generally, ototoxic drugs and very intense broad-band noise affect the basal portion of the cochlea first and, because of tonotopic organization, the ACCP responses to high frequency pure tones are affected before those to the low frequencies. However, the correlation between the effect of an ototraumatic agent on the ACCP and its effect on cochlear morphology is not always reliable. The correlations between changes in N1 and EP and in cochlear morphology are even less precise. Also discussed will be the cochlear effects of noise and the ototoxic interactions between drug/drug, noise/drug, and noise/drug/otitis media.

Acetylcysteine↗

Scanning electron microscopy of the cochlea in rats with Streptococcus pneumoniae otitis media.

This investigation was performed to determine the morphologic effects of bacterial otitis media on the organ of Corti. The middle ear cavities of rats were inoculated with Streptococcus pneumoniae or saline and the animals were killed on days 1, 4, 7, 10, 14, and 21 after inoculation. Middle ear cultures were obtained and the cochleas were examined using scanning electron microscopy. All animals killed on day 1 had positive cultures, but by day 21, all cultures were negative. Cochlear changes observed were (1) damage to supporting cells (Deiters' cells), (2) morphologic changes of hair cell stereocilia, and (3) loss of inner and outer hair cell stereocilia and cell bodies (to a lesser extent), especially in the lower middle and basal turns. These changes appeared to occur in a definite sequence; ie, damage to the supporting cells, changes in stereocilia, and, finally, hair cell loss. These data show that cochlear damage and hair loss can be associated with bacterial otitis media.

Animals↗

Impairment in cochlear function produced by chloramphenicol and noise.

Ototoxic interaction between chloramphenicol and noise was studied in two separate investigations. In the first study, permanent ototoxicity was demonstrated in a group of rats which were subjected to short-duration, high-intensity noise and were then given chloramphenicol orally. The anatomical damage in this group was consistent with observed changes in cochlear round window recordings of cochlear microphonics at 4 kHz and of the N1 component of the eighth nerve action potential. In the second study, a temporary depression in the function of the cochlea was observed in rats subjected to the noise-chloramphenicol regimen used in the first study. Depressions in recordings of the round window similar to those in the first study were seen only during the first five days of recordings. After the fifth day, the recordings of the round window were normal, indicating recovery from a temporary shift in threshold produced by chloramphenicol and noise. Incidence of purulent otitis media was found in 57 and 0% of the animals in the first and second studies, respectively. The combination of chloramphenicol and noise appears to be responsible for the production of temporary cochlear deficits. The addition of the third variable, otitis media, appears to result in permanent impairment of the cochlea.

Animals↗

Ototoxic drugs and noise.

Drugs that produce tinnitus can be subdivided into those which produce temporary or permanent hearing loss and those which apparently do not cause any hearing loss. The tinnitus occurring with drugs of the first group is probably secondary to the hearing loss. However, most of the drugs that produce tinnitus without an accompanying hearing loss probably do so because of their effect on biogenic amines in the central nervous system and/or as an extension of their proconvulsant side-effects. A pre-existing cochlear impairment is the underlying factor in most patients who experience tinnitus. Not only can ototoxic drugs or high levels of noise produce cochlear impairment but the interaction of the two can place humans in more jeopardy than when exposed to either agent alone. Chloramphenicol has little ototoxic potential when administered systemically in humans. However, our studies show that when chloramphenicol is combined with noise exposure in rats, considerably more cochlear damage results than from the noise alone (chloramphenicol alone does no produce any cochlear damage). We are presently conducting more detailed studies of this ototoxic interaction to determine whether it occurs with other antibiotics (such as erythromycin) which are also commonly considered to have minimal ototoxicity.

Acoustic Stimulation↗

Iron deficiency anemia and hearing.

Iron deficiency anemia is a frequently occurring clinical disorder. Despite the suggested association with hearing loss in the literature, cochlear sequelae of iron deficiency have yielded conflicting results in experimental studies. Auditory function was tested in iron-deficient and normal male Wistar albino rats using distortion product otoacoustic emissions and auditory brainstem response audiometry for the clarification of the opposing results in the literature. Hemoglobin, hematocrit, serum iron and albumin levels were monitored to verify iron deficiency. Although dramatic differences in weight gain and blood test parameters were noted, no significant change in auditory function due to iron deficiency was detected.

Anemia, Iron-Deficiency↗