The effects of isoflurane on 22Na+ influx, 45Ca2+ influx, catecholamine secretion and cyclic GMP production induced by three kinds of secretagogue (nicotinic agonists, veratridine and a high concentration of K+) have been investigated using cultured bovine adrenal medullary cells. (1) Isoflurane (1-6%) inhibited catecholamine secretion stimulated by carbachol, nicotine and dimethyl-4-phenylpiperazinium in a concentration-dependent manner. Isoflurane suppressed carbachol-evoked 22Na+ influx and 45Ca2+ influx at concentrations similar to those which suppressed catecholamine secretion. The inhibition of catecholamine secretion by isoflurane was not overcome by increasing the concentration of carbachol. (2) The inhibitory effects of isoflurane on veratridine-induced 22Na+ influx, 45Ca2+ influx and catecholamine secretion became evident when the concentration of isoflurane was raised to 4-6%, i.e. 2-3 fold higher than the concentrations (1-2%) employed clinically. (3) High K(+)-evoked 45Ca2+ influx and catecholamine secretion were not affected by isoflurane (1-6%). (4) Isoflurane (1-6%) attenuated the production of cyclic GMP caused by muscarine, but not that caused by atrial natriuretic peptide or by sodium nitroprusside. These results suggest that isoflurane, at clinical anesthetic concentrations, inhibits nicotinic acetylcholine receptor-mediated cell responses as well as muscarinic receptor-mediated cyclic GMP production in adrenal medullary cells.

Adrenal Medulla↗

PubMed1994

Differential up-regulation of voltage-dependent Na+ channels induced by phenytoin in brains of genetically seizure-susceptible (E1) and control (ddY) mice.

We investigated the effect of in vivo administration of an antiepileptic drug, phenytoin, on the saxitoxin binding capacity of receptor site 1 of the Na+ channel alpha-subunit, and the expression activity of the channel messenger RNA in epileptic El mouse brains, as compared with parental ddY mice. Subchronic treatment with phenytoin (25 mg/kg per day) for 14 days increased the [3H]saxitoxin binding to brain-derived synaptic membranes of both El and control ddY mice in a time dependent manner. This increase plateaued at 21 +/- 4% in El mice and 28 +/- 3% in ddY control mice after administration of phenytoin for seven days. After cessation of treatment with phenytoin, [3H]saxitoxin binding capacity returned to the basal level within two weeks in both ddY and El brains. Scatchard plot analysis revealed that the phenytoin treatment caused a 20-30% increase in maximum binding capacity of [3H]saxitoxin binding without any change in equilibrium dissociation constant in the brain cortical synaptic membranes of both epileptic El and control ddY mice. A single injection of phenytoin (25 mg/kg) elevated the level of Na+ channel messenger RNA within 1 h in ddY mouse brains. The increase in Na+ channel messenger RNA reached a peak (about 80% increase) after 5 h of phenytoin administration in a concentration-dependent manner (6.25-50 mg/kg). On the other hand, in El mouse brains, Na+ channel messenger RNA was not elevated until more than 5 h after phenytoin injection, and was increased by only about 33%.(ABSTRACT TRUNCATED AT 250 WORDS)

Amphibian Proteins↗

PubMed1994

The receptor, metabolism and effects of androgen in osteoblastic MC3T3-E1 cells.

We investigated the androgen receptor (AR), metabolism and effects of androgens in osteoblastic MC3T3-E1 cells. AR was proved as a transcript of a 10-kb mRNA and as a 110-kDa protein. An immunocytochemical study showed that AR was located mainly in the nuclei. Specific binding of [3H]DHT was observed in both the nuclear and cytosol fractions. MC3T3-E1 cells possessed approximately 1190 binding sites per cell and most of the sites (1150 sites) situated in the nucleus. The apparent Kd value in the nuclear fraction was 1.35 nM for [3H]DHT binding, and it was similar to that for [3H]testosterone. In the competition analysis, there was not much difference in the displacement of the [3H]DHT binding from AR between the addition of radioinert DHT and testosterone. In studies of 5 alpha-reductase activity and aromatase activity of the cells, both activities were lower than the respective values in classical androgen target tissues. Androgens stimulated the incorporation of [3H]thymidine into the cell, and DHT and testosterone had a similar potency on the cell proliferation. Thus, these results suggest testosterone itself acts mainly on the osteoblasts without conversion to DHT.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

PubMed1994

Resection of persistent nasopharyngeal carcinoma.

Although radiotherapy has been generally accepted as the treatment of choice for nasopharyngeal carcinoma (NPC), NPC at the primary site is not always controlled by this therapy. We performed surgical treatment to eradicate such residual tumor through a transmandibular, transpterygoid approach, on four patients with local residual NPC after curative radiotherapy. Two patients survived with no evidence of disease for more than 6 years. The other two patients died of multiple metastases to the liver or bone at 4 years and at 6 months after treatment. However, the nasopharynx remained free of disease in these two patients. Although the number of patients in the present series is small, surgical treatment of local persistent tumor after radiotherapy was beneficial. The indication for this technique is tumor in the nasopharynx extending to the parapharyngeal space, not invading intracranially. The transmandibular, transpterygoid approach offers a wide operative field with minimal postoperative morbidity, making it possible to manage the internal carotid artery easily.

Journal Article↗

PubMed1994

Regeneration of facial nerve after hypoglossal facial anastomosis: an animal study.

Hypoglossal-facial nerve anastomosis was carried out in 20 adult guinea pigs. Electromyographic responses of orbicularis oculi muscle evoked by blink reflex were recorded 2, 4, and 6 months after surgery. Then the anastomotic site was reopened, and a segment of buccal branch and the anastomotic trunk were resected for detailed histologic study. Regenerated axons were counted and the cross-sectional area of axons and fasciculi was measured. Data obtained from both blink reflex measurement and histologic study demonstrate a good quality of regeneration of the facial nerve from the hypoglossal nerve completed 6 months after the operation. In addition a new nerve bundle was regenerated from the proximal stump of the facial nerve connecting to the anastomotic site in 80% of the animals. Postoperative change in innervation pattern of the facial nerve was also illustrated.

Anastomosis, Surgical↗

PubMed1994

Experimental cholesteatomas arising from autologous free skin grafting in the middle ear cavity.

An autologous free skin graft taken from the upper or the lower external ear canal or the auricle was implanted in the otic bulla of 28 guinea pigs. An epidermal cyst simulating middle ear cholesteatoma was successfully produced in 25 (89.3%) animals 8 weeks after grafting. Using this model, the histological reaction of middle ear granulation tissue to the debris was studied. Cysts originated from the auricular skin showed the strongest activity with regard to growth rate, amount of debris produced, and inflammatory reaction to the exposed debris. We conclude that the exposed debris plays an important role in chronic granulomatous inflammation in association with cholesteatoma.

Animals↗

PubMed1994

Change in facial nerve innervation following hypoglossal-facial anastomosis. An animal study.

Hypoglossal-facial nerve anastomosis was carried out in 20 adult guinea pigs. Horseradish peroxidase (HRP) was applied to the buccal branch of the facial nerve at intervals of 2, 4 and 6 months after the anastomosis operation. HRP labeled neurons and the number of regenerated axons in the buccal branch were counted to determine postoperative nerve regeneration. The number of surviving neurons in the facial and hypoglossal nuclei was also counted to determine postoperative change of these nuclei. Following anastomosis, 97% of the hypoglossal neurons remained surviving, while the facial neurons underwent pronounced degeneration of 65% survival ratio at 2 months and 37% at 6 months after the anastomosis operation. In 80% of the animals, a new nerve bundle regenerated from the proximal stump of the facial nerve to the anastomotic trunk. A linear increase of HRP-labeled neurons in the facial and hypoglossal nuclei paralleled the increase of the axons in the buccal branch. The HRP-labeled neurons in the facial nucleus were demonstrated to have direct connection with the newly formed bundle and the facial mimetic muscles were dually innervated by both the hypoglossal and facial nerves. Although the present study design might not fully represent the clinical situation, possible advantages of hypoglossal-facial nerve anastomosis are discussed from the view of nerve regeneration.

Anastomosis, Surgical↗

PubMed1994

Effects of combined rupture of Reissner's membrane and the round window on hearing in the guinea pig.

Hearing impairment caused by sequential rupture of Reissner's membrane and the round window was investigated in guinea pigs to determine if the double-membrane break can cause acute profound sensorineural hearing loss. Reissner's membrane was lacerated through the stria vascularis at one of the turns of the cochlea, followed by rupture of the round window to create leakage of the perilymph. Action potentials (APs) to tone pip stimuli were recorded from the Fallopian canal electrode before and 90 min after these procedures. The results showed that the ears with the double-membrane break at the second turn showed a larger increase in AP threshold than did the control ears with only round window rupture. However, this was not seen when Reissner's membrane had been ruptured at the other turns.

Acoustic Stimulation↗

PubMed1994

Postoperative recurrence of perilymphatic fistulas.

Postoperative follow-up study of perilymphatic fistulas (PLFs) showed that recurrence of PLF was not rare and revision was sometimes needed to relieve the symptoms associated with leakage of perilymph. Of the 54 PLF patients surgically treated in our clinic, some sign or symptom of recurrence was found in 9 cases (17%). Vertigo accompanied by spontaneous or positional nystagmus was noted in all 9 cases, while only 3 complained of exacerbation of the existing hearing loss. Revision was indicated in 3 patients since they had no predisposition to spontaneous healing. Various etiological and underlying factors contributed to the incidence of recurrence. Careful operative procedures together with strict postoperative management are required for surgical treatment of PLF.

Adolescent↗

PubMed1994

Detection of human alpha-herpesvirus DNA using consensus primers and specific probes.

A simple and rapid diagnostic assay system was developed for detecting and identifying human alpha-herpesviruses, such as herpes simplex virus types 1 and 2 and varicella-zoster virus, by polymerase chain reaction. This system was based on an amplification step, using primers that bind the DNA polymerase consensus sequence of alpha-human herpesviruses, and a detection step, using non-radioactive virus-specific probes. This method could be used to amplify any human alpha-herpesviruses, and each virus-specific probe was highly specific for identification of the amplified product. This system is readily applicable for implementation in the clinical laboratory.

Cytomegalovirus↗

PubMed1994

Correlation of activation of Ca2+/calmodulin-dependent protein kinase II with catecholamine secretion and tyrosine hydroxylase activation in cultured bovine adrenal medullary cells.

We have investigated the activation of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) in cultured bovine adrenal medullary cells. The activation was assayed as an increase in the Ca(2+)-independent (autonomous) activity of CaM kinase II, using the synthetic substrate Syntide-2. Incubation of cells with acetylcholine increased the Ca(2+)-independent activity in a time (20 sec to 5.0 min)- and concentration (10-300 microM)-dependent manner. These curves were closely correlated with those of catecholamine secretion and tyrosine hydroxylase activation. Removal of extracellular Ca2+ completely abolished the stimulatory effects of acetylcholine on the Ca(2+)-independent activity, as well as on catecholamine secretion and activation of tyrosine hydroxylase. Nicotine but not muscarine increased the Ca(2+)-independent activity as potently as did acetylcholine, and hexamethonium but not atropine completely blocked the acetylcholine-induced increase. In 32P-labeled cells, acetylcholine stimulated the phosphorylation of a 50-kDa protein that was immunoprecipitated with an anti-brain CaM kinase II antibody. These results suggest that acetylcholine stimulates CaM kinase II activity through nicotinic acetylcholine receptor-mediated influx of Ca2+ and that the activation of CaM kinase II is closely related to catecholamine secretion and tyrosine hydroxylase activation in cultured adrenal medullary cells.

Acetylcholine↗

PubMed1994

Hearing improvement following removal of acoustic neuroma with preoperative sudden deafness.

We report three cases of small intracanalicular acoustic tumor with sudden hearing loss (SHL). The tumors were totally removed through the middle cranial fossa approach, preserving both the facial and cochlear nerves between one and four months after the onset of deafness. From one to three months postoperatively hearing has improved significantly. Hearing improvement in acoustic tumor surgery is a new aspect to be studied more in the future. Although the data are insufficient to draw broad conclusions, surgical removal of small acoustic tumors is rational to improve hearing, reduce or eliminate tinnitus and prevent recurrent attacks of deafness or vertigo. Factors relevant to postoperative hearing improvement are considered.

Audiometry↗

PubMed1994

C-type natriuretic peptide stimulates catecholamine synthesis through the accumulation of cyclic GMP in cultured bovine adrenal medullary cells.

We examined the effects of C-type natriuretic peptide (CNP) on cyclic GMP production and catecholamine synthesis in cultured bovine adrenal medullary cells. 1) CNP increased intracellular cyclic GMP content in a concentration-dependent manner (10-1000 nM). 2) The cyclic GMP production induced by 1 microM CNP reached a 200-fold increase, and the effect of CNP was most potent among the natriuretic peptide family. 3) The CNP-induced cyclic GMP production was attenuated by endothelin (1 microM) and angiotensin II (0.1-1 microM). 4) When the cells were cultured with hypertonic NaCl medium, the CNP-induced cyclic GMP production was potentiated in a time (1-4 days)- and concentration (25-100 mM)-dependent manner. 5) CNP stimulated the synthesis of 14C-labeled catecholamines from [14C] tyrosine but not from [14C] dopa. The stimulatory effect of CNP on the 14C-labeled catecholamine synthesis was observed at the concentrations of 100 to 100 nM. 6) 8-Bromo cyclic GMP, a membrane-permeable cyclic GMP analog, and sodium nitroprusside, an activator of soluble guanylate cyclase, also stimulated the synthesis of 14C-labeled catecholamines from [14C]tyrosine, whereas C-ANF, a specific ligand for the ANP-C (clearance) receptor that does not increase cyclic GMP content, failed to stimulate the synthesis of 14C-labeled catecholamines. 7) CNP (1 microM) as well as 8-bromo cyclic GMP and sodium nitroprusside increased the activity of tyrosine hydroxylase in the cells. These results suggest that in the adrenal medulla, CNP is a potent agonist for cyclic GMP production, which is modulated by endothelin, angiotensin II and the hypertonic NaCl condition.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenal Medulla↗