Membrane enzyme systems responsible for the Ca(2+)-dependent phosphorylation of Ser(27), the independent phosphorylation of Tyr(10) and Tyr(7), and the dephosphorylation of these phosphorylated residues in the alpha-chain of H/K-ATPase.

H/K-ATPase preparations (the G1 membrane) from pig stomach contain both kinases and phosphatases and show reversible phosphorylation of Tyr(7), Tyr(10), and Ser(27) residues of the alpha-chain of H/K-ATPase. The Tyr-kinase is sensitive to genistein and quercetin and recognized by anti-c-Src antibody. The Ser-kinase is dependent on Ca(2)(+) (K(0.5) = 0.9 microM), sensitive to a PKC inhibitor, and recognized by antibodies against PKCalpha and PKCbetaII. The addition of 3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonic acid (CHAPS) caused a dramatic increase in the phosphorylation of added synthetic copolymer substrates and permitted the phosphorylation of maltose-binding proteins fused with the N-terminal domain of alpha-chains. The phosphotyrosine phosphatase was inhibited by vanadate. The phosphoserine phosphatase was inhibited by okadaic acid and by inhibitor-2. The presence of protein phosphatase-1 was immunologically detected. Column chromatographic separation of CHAPS-solubilized G1 membrane and others indicate the apparent molecular weight of the Src-kinase to be approximately 60 kDa, the PKCalpha and/or PKCbII to be approximately 80 kDa, the Tyr-phosphatase to be 200 kDa, and PP-1 to be approximately 35 kDa. These data show that these membrane-bound enzyme systems are in sufficiently close proximity to be responsible for reversible phosphorylation of Tyr(7), Tyr(10), and Ser(27) of the catalytic subunit of membrane H/K-ATPase in parietal cells, the physiological role of which is unknown.

Intestinal cell kinase (ICK) localizes to the crypt region and requires a dual phosphorylation site found in map kinases.

Identification of key regulatory kinases in the intestinal epithelium are useful to understand the molecular mechanisms that underlie proliferation and differentiation in cells found in this compartment. We used the polymerase chain reaction (PCR) to amplify the catalytic kinase domain of serine-threonine kinases by employing degenerate primers and then screened an intestinal crypt cDNA library to clone and sequence the open reading frame of a novel serine-threonine kinase. This was then further characterized by Northern blot analysis and RNA in situ hybridization. This kinase, designated intestinal cell kinase, harbors a dual phosphorylation site found in mitogen-activating protein (MAP) kinases that is important for kinase activity.

Influence of alcohol on respiratory disturbance during sleep.

Oxygen saturation was measured in 37 patients with sleep-related breathing disorders over 2 nights: after alcohol intake and under control conditions. Both the number of 3% oxygen desaturation per hour (ODI3) and the lowest saturation (LSAT) were significantly aggravated after alcohol ingestion. Oxygen saturation was degraded in 28 cases after alcohol intake. In 69% of the cases in which ODI3 increased after alcohol ingestion, the aggravation during the alcohol metabolism time was noted. No correlation was found between the quantity of alcohol and oxygen desaturation following alcohol ingestion. Our study revealed that alcohol aggravates sleep-related breathing disorders.

The influence of sleep position and obesity on sleep apnea.

The influence of sleep position and the degree of obesity were examined in 257 subjects with sleep apnea. Subjects were divided into three groups according to obesity: normal weight (body mass index (BMI) under 24.0 kg/m2), mild obese (BMI 24.0-26.4 kg/m2) and obese group (BMI 26.4 kg/m2 and heavier). The apnea + hypopnea index (AHI), the intraesophageal pressure and the lowest oxygen saturation became significantly worse according to the degree of obesity. The subjects were also divided into two groups according to the reduction in the AHI by lateral position: good responders showed 50% or more reduction of AHI in lateral position and poor responders indicated less than 50% reduction. The percentage of good responders to sleep position change was 90.9% in normal weight group, 74.0% in mild obese group and 57.4% in the obese group. The ratio of the subjects who had indicated two or more obstructive sites in normal weight group was 36.0% in good responders and 40.0% in poor responders. The ratio in the mild obese group was 51.8% in good responders and 66.7% in poor responders. In the obese group, the ratio was 59.4% in good responders and 78.9% in poor responders.

Intensity of snoring in patients with sleep-related breathing disorders.

The intensity of snoring was measured in 73 patients with snoring or sleep apnea using a noise meter. The mean intensity of snoring at 50 cm in front of the mouth was 61.7 dB in the supine position and 53.7 dB in the lateral position. There existed a definite correlation between the logarithmic transformation of the intra-esophageal pressure amplitude and the intensity of snoring in the supine and lateral decubitus positions. These findings suggest that the intensity of snoring may be a useful index for sleep-related breathing disorders.

Evaluation of the auto continuous positive airway pressure efficacy by upper airway pressure measurement.

The efficacy of auto continuous positive airway pressure (CPAP) was prospectively evaluated in 11 obstructive sleep apnea cases by the upper airway pressure measurement. Their apnea plus hypopnea index decreased from 68.3+/-20.2/h to 0.5+/-0.9/h during manually adjusted and 9.6+/-14.5/h during auto CPAP The intraesophageal pressure decreased from -88.0+/-34.0 cm H2O to 11.9+/-7.2 cm H2O during manually adjusted and -26.0+/-13.7 cm H2O during auto CPAP. The manually adjusted CPAP level was 9.1+/-1.7 cm H2O and the maximum pressure of the auto CPAP was 9.4+/-2.0 cm H2O. Auto CPAP could automatically detect the optimal CPAP level. However, the efficacy of the auto CPAP was less than that of the manually adjusted CPAP.

Direct evidence for in vivo reversible tyrosine phosphorylation of the N-terminal domain of the H/K-ATPase alpha-subunit in mammalian stomach cells.

In vivo reversible phosphorylation of Tyr-7 and Tyr-10 of the pig stomach H/K-ATPase alpha-chain was initially demonstrated in mammals, rat, rabbit, and pig, in the presence of vanadate + H(2)O(2). In vitro phosphorylation has also been unequivocally demonstrated via the use of protease inhibitors during membrane H/K-ATPase preparation. An amphoretic detergent permitted each intrinsic kinase to phosphorylate each fusion protein containing the requisite Tyr residues, along with a reduction in alpha-chain phosphorylation. These and other data suggest that some important enzyme systems are present in the apical membrane and that they are in sufficient proximity to participate in the reversible phosphorylation of the amino terminal soluble domain of the alpha-chain with an unknown physiological function in the membrane embedded H/K-ATPase.

[Late phase II clinical study of RP56976 (docetaxel) in patients with advanced/recurrent head and neck cancer].

A late phase II clinical study of RP56976 (docetaxel), a new anticancer agent for advanced/recurrent head and neck cancer, was conducted in 29 institutions all over Japan as a multi-institutional cooperative study. Docetaxel was administered by 1 to 2-hour intravenous infusion at a dose of 60 mg/m2 every 3 to 4 weeks. Of 63 patients eligible in this study, 59 were judged as complete cases. Complete response (CR) was observed in 1 patient, partial response (PR) in 13, no change (NC) in 25, and progressive disease (PD) in 20, for an overall response rate of 22.2% (14/63, 95% CI: 12.7-34.5%) in eligible cases, and 23.7% (14/59, 95% CI: 13.6-36.6%) in complete cases. Previously treated patients showed a 17.9% (10/56) response rate, whereas treatment--naive patients showed a 57.1% (4/7) response rate. Among 46 patients who received prior chemotherapy, one CR and 7 PR were observed with a 17.4% response rate. Major hematological toxicities were leucopenia in 95.1% (> or = grade 3, 59.7%) and neutropenia in 90.3% (> or = grade 3, 79.0%). Other severe toxicities (> or = grade 3) included anorexia in 9.7% (6 cases), diarrhea in 3.2% (2 cases), dyspnea in 3.2% (2 cases), and fatigue in 3.2% (2 cases). One patient had a grade 3 interstitial pneumonia; however, symptoms were resolved by the administration of corticosteroids. During this study, one patient died due to multiple organ failure (MOF) caused by disseminated intravascular coagulation (DIC), and this case was reported as a therapy-related death. Based on these results, docetaxel is an active agent for treatment of head and neck cancer.

Effect of unilateral section of cervical afferent nerve upon optokinetic response and vestibular nystagmus induced by sinusoidal rotation in guinea pigs.

An experiment was carried out in guinea pigs to clarify the effect of unilateral section of cervical afferent nerve (C1-C3) upon optokinetic nystagmus and vestibular nystagmus induced by sinusoidal rotation. To produce optokinetic nystagmus and optokinetic after-nystagmus, a random dot pattern was utilized as visual stimulation at a speed of 30 degrees/s. As for vestibular nystagmus, sinusoidal rotation at a frequency of 0.1 Hz with an amplitude of 120 degrees was used. Results showed that for about a week after the surgical section of the C1-C3 nerves, directional preponderance of the vestibular nystagmus was found toward the lesion side, whereas no significant change was obtained in optokinetic nystagmus and optokinetic after nystagmus. This asymmetric change of the vestibular nystagmus was compensated for within a week or two. Thus. unilateral section of the cervical afferent nerve produced only a temporary effect on the vestibulo-ocular reflex but it had no significant effect on the optokinetic response.

p53 tumor suppressor gene in acoustic neuromas.

Alterations of the p53 tumor suppressor gene in 21 cases with acoustic neuroma were investigated by polymerase chain reaction-restriction fragment polymorphism (PCR-RFLP) and single strand conformation polymorphism (PCR-SSCP). Neither mutation nor deletion was found. In 13 informative cases, no loss of heterozygosity (LOH) was confirmed. Thus our results further substantiate the scant contribution of p53 gene tumorigenesis and cell proliferation to acoustic neuromas.

Significance of hearing preservation in acoustic neuroma surgery.

In the past 10 years, 43 patients with acoustic neuroma have been operated on by the middle cranial fossa approach. In all cases, meticulous care was taken to preserve the cochlear function regardless of the degree of preoperative hearing disorder. Thirty-nine of 43 patients had various degrees of residual hearing, hearing was preserved in 16 of these (41%). The best result was obtained in patients with a tumor located less than 1 cm from the porus. The hearing preservation rate was 64% (11/17). The patients recovered hearing acuity quite well, especially at low frequencies in 2 cases who had a pure tone hearing level of more than 50 dB and less than 50% of speech discrimination. These 2 cases had a history of sudden deafness that was intractable 5 months preoperatively. Thus it is not correct to set certain criteria for hearing preservation for patients whose preoperative hearing may not be serviceable.

Changes in nasal responsiveness to histamine and to specific antigen after laser surgery.

An initial treatment with several kinds of anti-allergic medicines is useful for reducing nasal allergy symptoms in patients suffering from Japanese cedar pollinosis during the pollen season. Since laser surgery before the pollen season seems to have a preventive effect as well, it would be of interest to know the time course of changes in the nasal reactivity to specific and non-specific stimuli after laser surgery. In this study, we investigated the changes in the nasal reactivities to specific antigen and histamine after CO2 laser surgery. The nasal reactivities to both specific antigen and histamine were enhanced 2 weeks after the laser surgery. On the other hand, they were significantly reduced after 4 weeks. Our data strongly suggest. therefore. that laser surgery must be done more than 4 weeks before the start of the pollen season to avoid temporary enhancement of nasal allergy symptoms.

Influence of nasal obstruction on obstructive sleep apnea.

Nasal obstruction, either partial or complete, may cause snoring and/or obstructive sleep apnea (OSA). However, there is no clear answer to the question, in how many cases nasal airway is a main or combined site of obstruction. Fifty-four OSA cases (47 men and 7 women) were examined by measuring the upper airway pressure gradient while they were asleep, using a multi-sensor pressure transducer. Their average apnea + hypopnea index was 35.6/h and body mass index was 26.5 kg/m2. Our hypothesis was that elevation of the inspiratory negative airway pressure at the epipharynx could indicate the influence of nasal obstruction. Seven cases out of 54 proved to have inspiratory negative pressure more than -3 cmH2O at the epipharynx during sleep. Four cases indicated elevated negative pressure in the lateral position, and 3 cases out of 19 CPAP candidates showed the same elevation in the supine position. In all 7 cases, the main obstructive site was either at the soft palate or the base of the tongue. Although nasal airway is the preferred route of respiration during sleep, nasal obstruction itself cannot be a main factor causing OSA, but a co-factor of OSA during sleep.

Acoustic analysis of snoring and the site of airway obstruction in sleep related respiratory disorders.

Seventy-five adult patients with sleep related respiratory disorders were examined by polysomnography with simultaneous recordings of the intraluminal pressure of the upper airway and snoring sound. Obstructed sites in the upper airway during sleep were determined by comparing the amplitude of respiratory fluctuation of the pressures in the epipharynx, mesopharynx, hypopharynx and esophagus. A definite correlation existed between the intensity of snoring sound and the amplitude of respiratory fluctuation of the intraesophageal pressure. Based on the results of the intraluminal pressure partitioning, the subjects were divided into the soft palate type (28), the tonsil/tongue base type (14), the combined type (27) and the larynx type (6). The average value of fundamental frequency (ff) was 102.8+/-34.9 Hz in the soft palate type, 331.7+/-144.8 Hz in the tonsil tongue base type, 115.7+/-58.9 Hz in the combined type and around 250 Hz in the larynx type.

Evaluation of uvulopalatopharyngoplasty in treatment of obstructive sleep apnea syndrome.

Many methods have been reported to select the proper candidates for uvulopalatopharyngoplasty (UPPP) among obstructive sleep apnea patients. Polysomnography with intraluminal pressure recordings in the upper airway was found useful to predict obstructive site(s) and hence the success of surgery. According to the pressure analysis, 11 cases with high negative pressure at the mesopharynx were selected for UPPP, while UPPP combined with midline partial glossectomy (UPPP-MLG) was done in 18 cases proved to have high negative pressure difference between the mesopharynx and esophagus. A reduction of the apnea hypopnea index (AHI) of more than 50% after UPPP was found in 8 cases (72.7%) and in 11 cases (61.l%) after UPPP-MLG. It was found that the response rate was lower among the second group if compared to that of the first because of other aggravating factors rather than simple obstruction at the level of the soft palate and or tonsils-tongue base level.

Late phase response in nasal mucosa closely correlated with immediate phase reaction and hyperreactivity to histamine.

It has been suggested that the onset of the late phase response (LPR) and hyperreactivity to non-specific stimuli occurs in the lower airway. However, its relationship in the nose has not yet been studied. This study was designed to examine the mechanism of LPR and the relationship between LPR and hyperreactivity. A total of 25 Japanese cedar pollinosis patients participated in this study. On the first visit, the frequency of sneezes, weight of nasal discharge, and the nasal airway resistance (NAR) were time-dependently measured without antigen challenge. The histamine reactivity was observed after 12 h. The same protocol was used during the second to fourth visits. The frequency of sneezes, weight of nasal discharge, and NAR were measured continuously for 12 h after antigen challenge, and nasal reactivity to histamine was observed. The percent change of NAR during immediate phase response (IR) and during LPR showed a significant correlation. The frequency of sneezes and weight of nasal discharge induced by histamine were both significantly higher in the positive than in the negative LPR group. These results suggest that the chemical mediators and inflammatory cells inducing nasal swelling during IR cause, directly or indirectly, nasal swelling during LPR, and induce hyperreactivity to histamine.

Effectiveness of tonsillectomy in adult sleep apnea syndrome.

Tonsillectomy was performed on 10 adult sleep apnea patients (five males, five females; average age, 39 years old; average body mass index, 24.8 kg/m2). Tonsillectomy alone was indicated if tonsillar hypertrophy was moderate to severe and the length of the soft palate was less than 35 mm according to cephalometry. Remarkable improvements were observed in all cases following surgery. The average weight of the resected tonsils was 11 g. The patients demonstrated a preoperative apnea + hypopnea index (AHI) of 14.4/h. This decreased to 2.9/h postoperatively. The intraesophageal pressure change improved from -36.6 to -15.7 cmH2O following surgery. A significant correlation between preoperative AHI and the degree of obesity (r=0.684, P < 0.05) was found. Although uvulopalatopharyngoplasty has been used extensively to treat sleep apnea, the present results suggests that certain subjects can be effectively treated through only tonsillectomy.