Relationship of cholera incidence to El Niño and solar activity elucidated by time-series analysis.

Using time-series analysis, we investigated the monthly cholera incidence in Dhaka, Bangladesh during an 18-year period for its relationship to the sea surface temperature (SST) linked to El Niño, and to the sunspot number. Dominant periodic modes identified for cholera incidence were 11.0, 4.8, 3.5, 2.9, 1.6, 1.0 and 0.5 years. The majority of these modes, e.g. the 11.0-, 4.8-, 3.5-, 1.6- and 1.0-year modes, were essentially consistent with those obtained for the SST data (dominant modes: 5.1, 3.7, 2.5, 2.1, 1.5, 1.0 years) and the sunspot number data (dominant modes: 22.1, 11.1, 7.3, 4.8, 3.1 years). We confirmed that the variations of cholera incidence were synchronous with SSTs, and were inversely correlated to the sunspot numbers. These results suggest that the cholera incidence in Bangladesh may have been influenced by the occurrence of El Niño and also by the periodic change of solar activity.

Gastric carcinoma confined to the muscularis propria: how can we detect, evaluate, and cure intermediate-stage carcinoma of the stomach?

OBJECTIVE: The most important surgical strategy for advanced gastric cancer is its detection at the curative stage. The aim of this study was to characterize the curable intermediate-stage gastric carcinomas. METHODS: Of 1120 consecutive patients who underwent gastric resection for primary gastric cancer from 1979 through 1996, 94 patients were histologically diagnosed as having cancer confined to the muscularis propria (mp cancer), analyzed clinicopathologically, and compared with patients with early and serosal cancers. RESULTS: The operative incidence of mp cancer was around 8% among cases of gastrectomy, and the ratio of mp cancer to advanced cancer began to increase in 1991. Mp cancer was at a statistically intermediate stage, between early and serosal cancers in terms of symptoms, surgical curability (96%), size and histology of the tumor, and the rate of lymph node metastasis (46%). Preoperative assessments of tumor depth were unclear using radiology and endoscopy; however, 35% of 31 cases studied were diagnosed precisely by endoscopic ultrasonography (EUS). Accuracy of lymph node metastasis diagnosis was the same (65%) by preoperative EUS and by surgeon; however, sensitivity of the surgeon's assessment was higher (69% vs 38%) and specificity of EUS was higher (83% vs 39%). The 5-yr survival rate was 85%, which was significantly better than that of serosal cancer and similar to that of early cancer. Patient outcome was not affected by lymph node metastasis or macroscopic type of tumor. CONCLUSIONS: Mp cancer should be considered an intermediate-stage cancer. Surgery with level 2 lymph node dissection should provide a cure rate similar to that for early cancer.

Analysis of rapid eye movement periodicity in narcoleptics based on maximum entropy method.

We examined REM sleep periodicity in typical narcoleptics and patients who had shown signs of a narcoleptic tetrad without HLA-DRB1*1501/DQB1*0602 or DR2 antigens, using spectral analysis based on the maximum entropy method. The REM sleep period of typical narcoleptics showed two peaks, one at 70-90 min and one at 110-130 min at night, and a single peak at around 70-90 min during the daytime. The nocturnal REM sleep period of typical narcoleptics may be composed of several different periods, one of which corresponds to that of their daytime REM sleep.

Relaxant effect of magnesium and zinc on histamine-induced bronchoconstriction in dogs.

OBJECTIVE: Magnesium sulfate (MgSO4) has been reported to produce bronchodilation in asthmatic patients. In vitro studies have suggested that divalent cations inhibit L-type voltage-sensitive calcium ion (Ca2+) channels in cardiac and smooth muscles. In this study, we evaluated the in vitro and in vivo effects of magnesium ion (Mg2+) and zinc ion (Zn2+) on the airway contracted by histamine. SETTING: A university research laboratory. IN VITRO: Tracheal smooth muscle from guinea pigs. IN VIVO: Mongrel dogs. IN VITRO STUDY: The tension of isolated guinea pig tracheal strips was measured isometrically with a force displacement transducer. The specimen was contracted with histamine (10 microM). Then, MgSO4 (n = 6), zinc sulfate (ZnSO4, n = 6), or sodium sulfate (Na2SO4, n = 6) was cumulatively added to the organ bath. IN VIVO STUDY: The bronchial cross-sectional area of mongrel dogs was measured by a direct visualization method demonstrated previously. The dogs were randomly assigned to three groups: group Mg (n = 7), group Zn (n = 7), and group Na (n = 7). Bronchoconstriction was elicited with histamine (10 microg/kg plus 500 microg/kg/hr iv). Thirty minutes after the start of histamine infusion, 0 (saline), 1, 10, and 100 micromol/kg ZnSO4 or 1, 10, 100, and 1000 micromol/kg MgSO4 or Na2SO4 were administered intravenously in group Zn, Mg, or Na, respectively. The bronchial cross-sectional area was assessed before (basal) and 30 mins after the start of histamine infusion and 5 mins after each dose of ZnSO4, MgSO4, or Na2SO4. Arterial blood was also obtained to measure plasma levels of epinephrine and norepinephrine by gas chromatography-mass spectrometry. All data are expressed as mean +/- SEM. The doses of the divalent cations that reversed histamine-induced contraction by 50% were calculated by GraphPad Prism. MgSO4 and ZnSO4 (9.38+/-0.28 and 1.84+/-0.30 mM, respectively) relaxed histamine-contracted tracheal strip in a concentration-dependent manner, whereas Na2SO4 did not. Similarly, the in vivo study showed that MgSO4 and ZnSO4 dose-dependently reversed histamine-induced bronchoconstriction (potency, ZnSO4 > MgSO4), whereas Na2SO4 did not. In groups Mg and Zn, the plasma catecholamine levels also dose-dependently increased except when 1000 micromol/kg MgSO4 was administered. CONCLUSION: Because the divalent cations tested produced a spasmolytic effect on the contracted airway, infusion of divalent cations might be effective against asthmatic attack. However, high concentrations of these cations produce significant toxicity, so dosage will be an important concern in development of these agents.

Prostaglandin E1 produces spasmolytic effects on histamine-induced bronchoconstriction in dogs.

OBJECTIVE: In this study, we evaluated the spasmolytic effect of intravenous prostaglandin (PG) E1 on histamine-induced bronchoconstriction with a direct visualization method using a superfine fiberoptic bronchoscope. SETTING: A university research laboratory. SUBJECTS: Mongrel dogs. INTERVENTIONS: The bronchial cross-sectional area (BCA) of mongrel dogs was measured by a direct visualization method using a superfine fiberoptic bronchoscope. Bronchoconstriction was elicited with histamine (H) infusion: 10 microg/kg iv bolus + 500 microg/kg/h continuous iv. The first protocol (n = 7) was designed to determine the effects of intravenous bolus of PGE1: 0 (saline), 0.01, 0.1, 1.0 and 10 microg/kg on H-induced bronchoconstriction. BCA was assessed before and 30 min after starting the H infusion and 5 min after each dose of intravenous PGE1. The second protocol was designed to determine whether continuous intravenous infusion of PGE1 reverses H-induced bronchoconstriction. In the PG group (n = 6), PGE1 was continuously infused at 0.1 microg/kg/min (20 mL/hr). In the control group (n = 6), saline was administered at a rate of 20 mL/hr iv. BCA was assessed before and 30 min after starting the H-infusion and at 5, 10, 30 and 60 min after commencing the PGE1 or saline infusion. Arterial blood was obtained simultaneously for measurement of plasma concentrations of epinephrine and norepinephrine by gas chromatography mass spectrometry. MEASUREMENTS AND MAIN RESULTS: In the first protocol, PGE1 produced a dose-dependent increase in the percentage of BCA and 10 microg/kg of PGE1 almost fully reversed the H-induced bronchoconstriction. Plasma catecholamines did not change significantly. In the second protocol, continuous infusion of PGE1 produced a time-dependent reversal of H-induced bronchoconstriction (percentage of BCA increased to 80.0+/-9.0% 60 min after the start of PGE1 infusion), whereas saline infusion did not reverse the bronchoconstriction. Plasma catecholamines did not change significantly in either group. CONCLUSIONS: Both intravenous bolus and continuous intravenous infusion of PGE1 reversed the H-induced bronchoconstriction. PGE1 may be used safely for patients with the hyperreactive airway and might be useful as a therapeutic agent for these patients.

Effects of thiopental on airway calibre in dogs: direct visualization method using a superfine fibreoptic bronchoscope.

Induction of anaesthesia with thiopental sometimes causes bronchospasm. Although the mechanism by which thiopental induces bronchospasm may involve cholinergic stimulation, direct spastic effect and histamine release, the spastic effects of thiopental have not been comprehensively defined. In this study, we have assessed the effect of thiopental on in vivo airway smooth muscle tone using direct visualization method with a superfine fibreoptic bronchoscope as previously reported. Twenty-one mongrel dogs were anaesthetized with pentobarbital (30 mg kg-1) and paralysed with pancuronium (200 micrograms kg-1 h-1). The trachea was intubated with a tube that had a second lumen for insertion of the bronchoscope (od: 2.2 mm) to continuously measure bronchial cross-sectional area. The tip of the bronchoscope was placed between the second and third bronchial bifurcation of the right lung. The dogs were allocated to three groups of seven: group T, A+T, H+T. In group T, thiopental 0 (saline), 0.1, 1.0 and 10 mg kg-1 was given i.v. In group A+T, saline i.v., 5 min later atropine 0.1 mg kg-1 i.v., and 5 min later thiopental 10 mg kg-1 was administered. In group H+T, bronchoconstriction was produced with histamine 10 micrograms kg-1 i.v. followed by infusion at 500 micrograms kg-1 h-1. Thirty minutes later, thiopental 0, 1.0 and 10 mg kg-1 were given. Arterial blood sampling was performed for measurement of plasma catecholamines and histamine. In group T, thiopental significantly reduced bronchial cross-sectional area (maximally by 28.7 (5.6% at 0.5 min after thiopental 10 mg kg-1), which returned to the baseline in 3 min, while any changes in plasma concentrations of catecholamines and histamine were not observed except norepinephrine level at 1 min following thiopental 10 mg kg-1 i.v. Atropine pretreatment completely prevented thiopental-induced bronchospasm in group A+T. In group H+T, thiopental 10 mg kg-1 transiently but significantly decreases bronchial cross-sectional area. Therefore, the present study indicates that the mechanism of thiopental bronchospasm may result from cholinergic nerve stimulation.

Midazolam reverses histamine-induced bronchoconstriction in dogs.

PURPOSE: Midazolam has been used clinically as a sedative and as an anaesthetic induction agent. However, the bronchodilating effects of midazolam have not been comprehensively evaluated. We sought to determine relaxant effects of midazolam on the airway. METHODS: After our Animal Care Committee approved the study, eight mongrel dogs were anaesthetized with 30 mg.kg-1 pentobarbitone iv, and were paralysed with 200 micrograms.kg-1.hr-1 pancuronium. The trachea was intubated with an endotracheal tube (ID 7 mm) that had a second lumen for insertion of a superfine fibreoptic bronchoscope (OD 2.2 mm) to measure the bronchial cross-sectional area (BCA) continuously. The tip of the bronchoscope was placed at the level of the second or third bronchial bifurcation of the right bronchus. A videoprinter printed the BCA which was then measured with a NIH image program. Bronchoconstriction was produced with histamine (H) 10 micrograms.kg-1 followed by 500 micrograms.kg-1.hr-1. Thirty minutes later, 0 [saline], 0.01, 0.1 and 1.0 mg.kg-1 midazolam and 25 micrograms.kg-1 flumazenil were given. The BCA was assessed before (basal area) and 30 min after the start of H infusion, and was also measured five minutes after each midazolam and flumazenil iv. At the same time, arterial blood was sampled for plasma catecholamine measurement. RESULTS: Histamine infusion decreased BCA to 49.7 +/- 17.3% of basal BCA. More than 0.1 mg.kg-1 midazolam increased BCA up to 71.7 +/- 15.3% of the basal (1.0 mg.kg-1) (P < 0.01). Plasma adrenaline concentration was decreased from 6.9 +/- 3.8 to 3.7 +/- 1.9 ng.ml-1 by 1.0 mg.kg-1 midazolam (P < 0.05). Flumazenil did not antagonize the relaxant effect of midazolam but reversed the inhibitory effect of midazolam on histamine-induced adrenaline release. CONCLUSION: Midazolam has a spasmolytic effect on constricted airways but this bronchodilatation was not reversed by flumazenil.

New technique for time series analysis combining the maximum entropy method and non-linear least squares method: its value in heart rate variability analysis.

A new technique for time series analysis, which is a combination of the maximum entropy method (MEM) for spectral analysis and the non-linear least squares method (LSM) for fitting analysis, is described. In this technique, the MEM power spectral density (MEMPSD) is calculated using a very large lag that could diminish the lag dependence of dominant periods estimated by the MEM analysis. The validity of this large lag is confirmed by the LSM, given that the ten dominant MEM periods are known quantities. To validate the MEM plus LSM technique, it is compared with autoregressive (AR) modelling, by analysing heart rate variability under pharmacological interventions (phenylephrine and trinitroglycerine), using 16 young males. The results indicate that the MEMPSD, when compared with the ARPSD, has numerous periods that could reproduce the original time series much more accurately, as revealed by the LSM analysis. However, both the low- and high-frequency powers with MEMPSD and ARPSDs shift in the expected directions in accordance with the pharmacological effects on the cardiovascular system. The implications of these results are discussed from the theoretical and practical standpoints of the MEM plus LSM technique, compared with AR modelling.

In vivo direct measurement of the bronchodilating effect of sevoflurane using a superfine fiberoptic bronchoscope: comparison with enflurane and halothane.

OBJECTIVES: Although volatile anesthetics have been shown to have spasmolytic effects on constricted airways, most previous studies were performed to assess bronchodilation with indirect methods such as measurement of airway resistance, whose reliability is controversial. As the authors have developed a new direct method using a superfine fiberoptic bronchoscope (SFB) and confirmed the accuracy of this method, this study assessed the spasmolytic effect of sevoflurane (S) as compared with enflurane (E) and halothane (H). DESIGN: Open-labeled, randomized study. SETTING: The study protocol was approved by the Animal Care Committee in a school of medicine. PARTICIPANTS: Twenty-two mongrel dogs. INTERVENTIONS: The dogs were anesthetized with pentobarbital, IV, paralyzed with pancuronium, and the lungs were mechanically ventilated. The endotracheal tube had an additional lumen to insert the SFB (outer diameter 2.2 mm). The tip of the SFB was located between a second and third bronchial bifurcation to continuously monitor the bronchial cross-sectional area (BCA) of third or fourth generation bronchi. The BCA was printed out by videoprinter at the end of expiration and was calculated on a Macintosh IIci computer using an NIH (National Institutes of Health) image program. Bronchoconstriction was produced with histamine (HA: 10 micrograms/kg + 500 micrograms/kg/hr). In the first protocol, 4 mongrel dogs were used to assess the accuracy of the new method. BCA was measured 30 and 60 minutes after the placement of the SFB and also 30 and 60 minutes after the start of the histamine (HA) infusion. Arterial blood (4 mL) was collected at 30 and 60 minutes after placement of the SFB to measure the plasma concentration of catecholamines with gas chromatographic mass spectrometry. In the second protocol, 18 dogs were randomly allocated to 3 groups of 6 each: group S, group E, and group H. Thirty minutes after the start of the HA infusion, the dogs subsequently inhaled the following concentrations of S, E, or H for 15 minutes: 0.2, 0.4, 0.8, and 1.6 minimum alveolar concentrations. The BCA was assessed before and 30 minutes after the start of HA and at the end of the inhalation period of each concentration. MEASUREMENTS AND MAIN RESULTS: In the first study, no significant differences in the BCA were found between 30 and 60 minutes after the placement of the SFB and between 30 and 60 minutes after the start of HA. The plasma concentrations of norepinephrine and epinephrine changed from 138 +/- 45 and 162 +/- 18 to 188 +/- 48 and 136 +/- 18 pg/mL, respectively. In the second study, all volatile anesthetics significantly increased the BCA in a dose-dependent manner. There was no significant difference among the three groups. CONCLUSION: As the BCA did not change in the first study, the tip of the SBF could be well fixed, and the 30-minute interval is long enough for the HA infusion to produce stable bronchoconstriction. The plasma catecholamine concentrations suggest that the pentobarbital anesthesia was deep enough to inhibit the direct irritant effect of the endotracheal tube and the SFB on the airway. This new direct method indicates that S dilates HA-constricted proximal airway with the same potency as E and H.

[Preliminary screening for antiviral AIDS drugs. VI. Report for fiscal year 1993].

Preliminary screening of antiviral AIDS drugs has been carried out using three different in vitro assay systems. Among 138 samples tested, two were found to inhibit the growth of HIV in vitro. Neither of the positive samples has hopeful signs, as the ranges of effective doses of the samples are very narrow.

[Changes in rectal temperature during tympanoplasty under general anesthesia].

We measured rectal temperature of 35 patients who underwent tympanoplasty under various types of general anesthesia. They were allocated at random to three groups according to the types of general anesthesia; 10 patients of enflurane-N2O (enflurane group); 10 patients of neuroleptic anesthesia with droperidol, pentazocine-N2O (NLA group); 15 patients of total intravenous anesthesia with droperidol, fentanyl and ketamine (DFK group). After the induction of anesthesia, their rectal temperature was continuously monitored with an electric thermometer of NEC San-ei throughout the surgical procedure. Increase in rectal temperature was observed in all three groups, and increase in DFK group was significant compared with other two groups. It is possible that the significant increase in rectal temperature in DFK group is supposedly due to normally maintained hypothalamic thermoregulatory function as well as direct surgical stimulation to central nervous system.

[Clinical study on intraoperative hyperketonemia in non-diabetic surgical patients under general anesthesia].

We measured plasma 3-hydroxybutyrate (3-OHBA) concentrations in 925 non-diabetic surgical patients who underwent various surgical procedures under various types of general anesthesia. Lactated Ringer's solution only was used as an intraoperative fluid. Among them, 46 patients (4.9%) developed high 3-OHBA levels of over 500 microM.l-1 during surgery. The causes of observed hyperketonemia would be surgical stress and preoperative fasting irrespective of 8 types of general anesthesia. Blood glucose, lactic acid, base excess and pH were not influenced by hyperketonemia. Prolonged recovery from anesthesia was not observed in any patients with hyperketonemia. Without any specific treatments, plasma 3-OHBA levels were unchanged or rather decreased in 70% of the patients whose 3-OHBA levels were over 500 microM. The results suggest that hyperketonemia below 500 microM.l-1 is not always disadvantageous in nondiabetic patients under general anesthesia.

[Clinical study on total intravenous anesthesia with droperidol, fentanyl, and ketamine--18. Effect on peripheral circulation as judged by core-peripheral temperature gradient].

Effect of total intravenous anesthesia with droperidol, fentanyl and ketamine (DFK) on peripheral circulation was studied by examining core-peripheral temperature gradient in twenty five patients who underwent abdominal surgery. A core temperature probe was attached on the forehead and peripheral probe on the palm of the hand of the side on which the blood pressure cuff was not applied. The temperature gradient was less than three degrees centigrade in 60% of the patients and the gradient was significantly less as compared with that of isoflurane anesthesia even at 300 minutes after the start of surgical operation. This advantage would have been caused by such factors as circulatory stimulating effect of ketamine, sympathetic blocking effect by droperidol and adequate postoperative analgesia by fentanyl and norketamine, a metabolite of ketamine. The results suggest that DFK would exert a beneficial effect on peripheral circulation, particularly during prolonged surgical procedures.

Transforming activity of a synthetic c-Ha-ras gene containing O6-methylguanine in codon 12.

A mutagenic DNA-adduct, O6-methylguanine, was introduced into codon 12 of the synthetic c-Ha-ras gene by cassette mutagenesis. Transfection of this modified ras gene into normal NIH3T3 cells by the calcium phosphate procedure resulted in significant induction of focus formation. The ras gene inserted into the transformed cells was found to have a G to A transition at the position of the modified base. These results indicate that an O6-methylguanine residue in DNA may lead to a mutation and be one cause of activation of the ras gene.

[Preliminary screening for antiviral AIDS drugs. II. Report on fiscal year 1989].

Preliminary screening for antiviral AIDS drugs was carried out using three different in vitro assay systems. Among 104 samples tested, six were found to inhibit the growth of HIV in vitro. Four of six were acidic polysaccharides of sea weed origin and one was a well-known anti HIV chemical whose anti-HIV activity has not been reported.

Pathogenesis of Vibrio parahaemolyticus: intraperitoneal and orogastric challenge experiments in mice.

It has been noted that V. parahaemolyticus isolated from patients with food poisoning are almost always thermostable direct hemolysin (TDH)-positive, whereas, isolates from foods or environmental sources are usually TDH-negative. The virulence of V. parahaemolyticus in mice was examined by using intraperitoneal and orogastric challenge models, where the strains used were those isolated from patients and foods with food poisoning and included TDH-positive and -negative. The LD50 in mice was estimated to be approximately 10(7) in the former and 10(8) in the latter challenge model. In the mice killed by challenge, either intraperitoneal or orogastric, the following pathological changes were almost always observed: swelling, redness and fluid accumulation in the small intestine, particularly the upper part of it. Histologically, congestion, edema, and vacuolation were observed in mucosal and submucosal tissues; furthermore, ulceration at tips of some villi was noticed. The lethality and pathological finding in mice due to V. parahaemolyticus were demonstrated similarly in both challenge models and irrespective of the TDH phenotype of the strain used. It will be suggested that the findings in challenged mice may be attributed to enteropathogenic factor(s) other than TDH of V. parahaemolyticus.

[Preliminary screening for antiviral AIDS drugs. I. Report on fiscal year 1988].

Preliminary screening for antiviral AIDS drugs has been carried out, using three different in vitro assay systems. Among 105 samples tested, 13 were found to inhibit the growth of HIV in vitro. Eleven of 13 were well-known anti-HIV chemicals, while the remaining two of plant origin were new chemicals whose anti-HIV activities have not been reported elsewhere.

Transformation of NIH3T3 cells with synthetic c-Ha-ras genes.

Synthetic human c-Ha-ras genes in which amino acid codons were altered to those which are frequently used in highly expressed Escherichia coli genes were ligated to the 3'-end of Rous sarcoma virus long terminal repeat. When NIH3T3 cells were transfected with the plasmids having those genes with valine at codon 12, leucine at codon 61 or arginine at codon 61, transformants were efficiently produced. These results indicated that the synthetic c-Ha-ras genes are expressed in a mammalian system even though their codon usage is altered to correspond with that of E. coli. This expression vector system should be useful for studies on the structure-function relationships of c-Ha-ras, since the synthetic gene can be easily modified to have multiple base alterations, and can also be used simultaneously for the production of large amounts of p21 in E. coli for biochemical and biophysical studies.