Correlations of health-related quality of life questionnaire results in asthma and allergic rhinitis: effects of a leukotriene receptor antagonist.

Bronchial asthma and allergic rhinitis frequently coexist. This study investigated correlations of health-related quality of life (QOL) questionnaires for these diseases, assessing whether the selective leukotriene receptor antagonist (LTRA), pranlukast, had additional benefits to overall asthma control when there was concomitant allergic rhinitis. Patients with asthma-associated allergic rhinitis were randomly allocated to either LTRA(+) (n = 21, treated for 3 months with pranlukast), or LTRA(-) (n = 8, no pranlukast). At study start and at 3 months, pulmonary function was evaluated and QOL assessments were made using the Asthma Health Questionnaire-Japan (AHQ-Japan) and the Japan Rhino-conjunctivitis Quality of Life Questionnaire (JRQLQ). Total scores were significantly correlated both before and after therapy. After 3 months' therapy, pulmonary function and total AHQ-Japan and JRQLQ scores significantly improved in the LTRA(+) group, but not in the LTRA(-) group. A significant correlation between change at 3 months in the AHQ-Japan and JRQLQ scores from baseline values was seen in the LTRA(+) group. LTRA therapy improved allergic rhinitis symptoms, asthma symptoms and pulmonary function.

Can nutrition influence circadian rhythm and heart rate variability?

Recent studies indicate that there is an interaction between biorhythms, the biological clock and triggers, which may be important in the pathogenesis of altered heart rate variability (HRV) and blood pressure variability (BPV). Circadian rhythms are under the influence of, and physiological variables are mediated by the activation of the adrenals, sympathetic/parasympathetic, hypothalamic and pituitary activity. Emotional stress, physical exertion, sleep deprivation and large fatty meals are major triggers of myocardial ischemia, angina, infarction, sudden cardiac death (SCD) and stroke. These events have been reported to exhibit a circadian variation with increased frequency in the second quarter of the day, which has also been observed in our studies on Indians. Recent studies indicate that altered HRV and BPV are also important in the pathogenesis and progression of heart failure, atheroma and thrombosis. Mediation via beta-blockers, oestrogens, n-3 fatty acids, vitamin E and coenzyme Q10 and fasting appears to have a beneficial influence whereas progestins, nifedipine, stress and exercise may have an adverse effect on HRV and BPV. We have reported that plasma levels of vitamin E and C are lower in the second quarter of the day than at other times, indicating their role in the pathogenesis of variability and cardiac events. Prospective studies also indicate that HRV and BPV are important and independent risk factors for cardiovascular events. However, no study has yet been conducted in patients with abnormal HRV and BPV in a randomized, placebo-controlled intervention trial to find out whether improvement in variability can cause a significant reduction in cardiovascular events. There is a need to study the role of n-3 fatty acids, coenzyme Q10, the effect of regular physical training, medication and ACE inhibitors in patients with abnormal HRV and BPV to demonstrate that improving variability can modulate cardiovascular events.

Alternating light-darkness-influenced human electrocardiographic magnetoreception in association with geomagnetic pulsations.

Geomagnetic variations of partly interplanetary origin, with cyclic signatures in human affairs and pathology include the incidence of various diseases, regarding which this study of healthy subjects attempted to determine an underlying mechanism by worldwide archival and physiological monitoring, notably of heart rate variability (HRV). In the past half-century, the possible health and other hazards of natural, solar variability-driven temporal variations in the earth's magnetic field have become a controversial subject in view of the inconsistent results. Some well-documented claims of associations between geomagnetic storms and myocardial infarction or stroke have been rejected by a study based on more comprehensive data analyzed by rigorous methods - covering, however, only part of a solar cycle in only part of a hemisphere. It seems possible that inter-solar cycle and geographic variability, if not geographic differences, may account for discrepancies. Herein, we examine the start of a planetary study on any influence of geomagnetic disturbances that are most pronounced in the auroral oval, on human HRV. The magnetic field variations exhibit complex spectra and include the frequency band between 0.001-10 Hz, which is regarded as ultra-low frequency by physicists. Since the 'ultra-low-frequency' range, like other endpoints used in cardiology, refers to much higher frequencies than the about-yearly changes that are here shown to play a role in environmental-organismic interactions revealed by HRV, the current designations used in cardiology are all placed in quotation marks to indicate the need for possible revision. Whether or not this suggestion has an immediate response, we have pointed to a need for the development of instrumentation and software that renders the assessment of circadian, infradian and even infra-annual (truly low frequency) modulations routinely feasible. HRV was examined on the basis of nearly continuous 7-day records by ECG between December 10, 1998, and November 2, 2000, on 19 clinically healthy subjects, 21 to 54 years of age, in Alta, Norway. A geomagnetic record was obtained from the Auroral Observatory of the University of Tromsø. First, frequency-domain measures of HRV were compared for each person in 24-hour spans of high geomagnetic disturbance versus quiet conditions. Second, cross-spectra between geomagnetic activity and HRV measures were quantified via the squared coherence spectrum using 7-day time series. A 7.5% increase in the 24-hour average of heart rate, HR (P = 0.00020) and a decrease in HRV were documented on days of high geomagnetic disturbance. The decrease in HRV was validated statistically for the 'total frequency', 'TF' endpoint (18.6% decrease, P= 0.00009). The decrease in spectral power was found primarily in the 'circaminutan frequency', 'VLF' (21.9% decrease, P< 0.000001) in conjunction with the 'minutes-to-hours' component, ultra-low-frequency, 'ULF' (15.5% decrease, P= 0.00865) and circadecasecundan 'low frequency', 'LF' (14.2% decrease, P = 0.00187) regions of the spectrum. Power-law scaling of the power spectra did not show any statistically significant difference. It is noteworthy that most of the decrease in HRV, except for the circaminutan (VLF) component, was observed only in the season in which sunshine alternated with darkness (D/L), a finding suggesting a mechanism influenced by the alternation of light and darkness. The hypothesis of a light-dark-influenced magnetoreception was also supported by cross-spectral analysis. Group-averaged coherence at frequencies coincident with the geomagnetic Pc 6 pulsations (with periods ranging from 10 minutes to 5 hours) differed with a statistical significance (P < 0.000001) among the three natural lighting conditions, the association being weaker during UL or D/D than during D/L. By contrast, no statistically significant differences were found in terms of the circadian and circasemidian frequencies in relation to the alternation of sunshine with darkness or rather circannual rhythm stage. In conclusion, evidence is provided herein that an alteration of HRV is most apparent in the circaminutan ('VLF') region, which is clinically important, because a reduction in its power is a predictor of morbidity and mortality from cardiovascular disease. The circadecasecundan ('LF') component of HRV also decreased in association with geomagnetic disturbance, which may reflect an episodic alteration of arterial pressure related to changes in geomagnetic activity. Lastly, our study suggests the existence of a light-dark-influenced magnetoreception mechanism in humans involving mainly the Pc 6 band of the magnetic field.

The diurnal rhythm of energy expenditure differs between obese and glucose-intolerant rats and streptozotocin-induced diabetic rats.

Otsuka Long Evans Tokushima Fatty (OLETF) rats were developed as a model of noninsulin-dependent diabetes mellitus (NIDDM) with mild obesity. Changes in carcass composition and in the daily profile of energy expenditure were examined before and after manifestation of diabetes (8 and 24 wk, respectively), and compared with the normal control Long Evans Tokushima (LETO) rats and streptozotocin (STZ)-induced diabetic LETO rats. OLETF rats had greater body weights than LETO rats and significantly greater absolute and relative fat weights. A diurnal rhythm of energy expenditure associated with two peaks was observed in LETO rats, but the two peaks were not apparent in OLETF rats at 24 wk of age. A diurnal rhythm associated with one peak was observed in STZ-induced diabetic LETO rats. Energy derived from fat constituted this peak; the pattern of the daily energy expenditure was significantly different from that of either nontreated LETO or OLETF rats at 24 wk of age. NIDDM in OLETF rats at 24 wk of age has only a small role in modification of the diurnal rhythm of energy expenditure, whereas STZ-induced diabetes significantly affected the rhythm.

Effects of rolipram, a selective inhibitor of phosphodiesterase 4, on hyperlocomotion induced by several abused drugs in mice.

The effects of rolipram, a selective inhibitor of phosphodiesterase 4, on the hyperlocomotion induced by several abused drugs (methamphetamine, morphine and phencyclidine) and a dopamine D1-receptor agonist (SKF81297; (+/-)-6-chloro-7,8-dihydroxy-1-phenyl-2,3 ,4,5-tetrahydro-1H-3-benzazepin hydrobromide) in mice were investigated. Methamphetamine (0.5-2.0 mg/kg), morphine (5.0-20 mg/kg), phencyclidine (1.25-5.0 mg/kg) and SKF81297 (2.5-10 mg/kg) each induced dose-dependent hyperlocomotion. A low dose (1.0 mg/kg) or moderate dose (3.2 mg/kg) of rolipram suppressed methamphetamine (2.0 mg/kg)- and morphine (20 mg/kg)-induced hyperlocomotion, but not phencyclidine (5.0 mg/kg)-induced hyperlocomotion. These results suggest that cAMP in the brain is involved in methamphetamine- and morphine-induced hyperlocomotion, while the underlying mechanism(s) of phencyclidine-induced hyperlocomotion may be different from those of methamphetamine- and morphine-induced hyperlocomotion. It is well known that methamphetamine- and morphine-induced hyperlocomotion are mediated by the dopaminergic system and that interaction between postsynaptic D1- and D2-receptors may play an important role in the expression of various dopamine-mediated behaviors. In the present study, SKF81297 (10 mg/kg)-induced hyperlocomotion was significantly but not completely suppressed by the highest dose of rolipram (10 mg/kg). Therefore it is unlikely that postsynaptic D1-receptor-mediated functions are involved in the suppressive effects of rolipram on methamphetamine- and morphine-induced hyperlocomotion. These results suggest that rolipram may inhibit methamphetamine- and morphine-induced hyperlocomotion via increase cAMP levels at D2-receptors.

[Pharmacological and clinical aspects of fluvoxamine (Depromel), the first selective serotonin reuptake inhibitor approved for clinical use employed in Japan].

Fluvoxamine (Depromel), a selective serotonin reuptake inhibitor (SSRI), was launched in May 1999 in Japan with more than 10 years' delay from the marketing in Europe and the United States. Fluvoxamine has been approved in about 80 countries as the indication to "depression" since 1983. As the indication to obsessive-compulsive disorder (OCD), fluvoxamine was first approved in the United States in 1994 and then in about 30 countries. Efficacy of the drug on "depression and depressed state" was found to be comparable to traditional tricyclic antidepressants (TCAs) by the clinical studies in Japan. Indication to OCD was first approved for fluvoxamine in Japan. The antidepressant and the anti-OCD action are considered the result of the serotonin reuptake inhibition at the serotonergic neurons. Fluvoxamine has little affinity for muscarinic, adrenergic alpha 1- and histamine H1-receptors, which TCAs have. Therefore, fluvoxamine possesses less side effects such as dry mouse, disuria, dizziness, orthostatic hypotension and drowsiness, etc.; and it is useful for elderly patients and long-term treatments for depression and OCD.

Development of the polysomnographic database on CD-ROM.

We have developed a polysomnographic database on CD-ROM. The data were obtained from 16 subjects with sleep apnea syndrome. The physiological signals include electroencephalogram, electromyogram, electrooculogram, invasive blood pressure, respiratory wave, oxygen saturation, and cardiac volume as measured by VEST method. The CD-ROM also include programs to analyze polysomnography (PSG) data. The CD-ROM has values: (i) for researchers investigating clinical physiology or non-linear dynamics during sleep apnea syndrome; (ii) for engineers developing a new algorithm for the computerized analysis of PSG data related to sleep apnea syndrome; (iii) for students learning sleep physiology.

Development of an analysis system for 24-hour blood pressure and heart rate variability in the rat.

We have developed a system to evaluate 24-h changes of heart rate (HR) and blood pressure variability (BPV) in free moving rat. As a sensor to monitor electrocardiogram, we used the telemetry system. Analog data were sent to an analog to digital converter directly. The data were digitized at a sampling frequency of 1250 Hz and digitally stored on a computer. RR-intervals (difference of successive RR in electrocardiogram) were calculated using custom designed programs. Circadian change of the blood pressure and heart rate variability were calculated. We suggest that the system is a powerful tool in the evaluation of circadian changes of hemodynamics and autonomic nervous function in free moving rats.

[Cardiovascular diseases].

Death from heart disease is sometimes observed at night. Life threatening arrhythmias or ischemic heart disease are suspected to be the cause of sudden death during night. Cheyne-Stokes respiration (CSR) is frequently observed in patients with chronic cardiac failure. CSR augments sympathetic nervous activity and reduces the quality of sleep. Sleep apnea or snoring is another stressful condition during sleep. During hyperventilatory phase of sleep apnea, the blood pressure, heart rate, end-systolic ventricular volume and vosomotor tone increases, and the periodic EEG arousal patterns are observed. Sleep apnea is suspected to be one of the risk factors of hypertension. The detection and early treatment of sleep apnea or Cheyne-Stokes respiration are required to reduce the mortality due to cardiac events during sleep.

Vitamin B12 treatment for delayed sleep phase syndrome: a multi-center double-blind study.

The active form of vitamin B12 (methylcobalamin) has been reported to be effective on sleep-wake rhythm disorders. Previous studies, however, were performed under open trial, and the effect of vitamin B12 has not been properly evaluated. The aim of this double-blind study was to investigate the efficacy of methylcobalamin on delayed sleep phase syndrome (DSPS). Methylcobalamin (3 mg/day) or placebo was administered for 4 weeks. The subjects were 50 patients with DSPS aged 13-55 years (26.8 +/- 1.3), 27 of whom received the active drug while 23 received the placebo. No significant differences were observed between the 2 groups in subjective evaluations of mood or drowsiness during the daytime or in night sleep by sleep-log evaluation. These results indicate that 3 mg methylcobalamin administered over 4 weeks is not an effective treatment for DSPS.

Amphetamine sensitization augments amphetamine-induced Fos expression in the lateral habenula.

Repeated amphetamine (AMPH) administration results in behavioral sensitization. To investigate the neuroanatomical basis of this phenomenon, we examined the effects of AMPH sensitization on AMPH-induced Fos expression in 24 regions of the rat brain. Rats received repeated injections of AMPH (4 mg/kg, intraperitoneally, once every other day, eight times in total) or saline (same schedule as for AMPH). After a 14-day drug abstinence period, rats were challenged with 2 mg/kg AMPH intraperitoneally. As measured by Fos immunohistochemistry, the AMPH sensitization procedure enhanced subsequent AMPH-induced Fos expression in only one structure, the medial part of the lateral habenula. These results indicate that AMPH-induced behavioral sensitization is not accompanied by widespread increases in the ability of AMPH to increase regional Fos expression in the forebrain. The lateral habenula appears to be involved in the possible neural framework that is responsible for the expression of behavioral sensitization.

Ameliorating effects of rolipram on experimentally induced impairments of learning and memory in rodents.

The effects of rolipram, a cAMP-specific phosphodiesterase (phosphodiesterase 4) inhibitor, on experimentally-induced amnesia were examined using a 3-panel runway paradigm in rats and a passive avoidance task in mice. Scopolamine, cerebral ischemia induced by four-vessel occlusion and electric convulsive shock impaired working memory in the 3-panel runway task. Rolipram at 0.1 mg/kg reduced the increase in errors induced by scopolamine or cerebral ischemia. Rolipram at 0.32 mg/kg also reduced the increase in errors induced by electric convulsive shock. Dibutyryl cAMP also had similar effects in 3-panel runway experiments. In the passive avoidance task, rolipram reversed the impairments of the avoidance response induced by scopolamine, cycloheximide and electric convulsive shock at 10, 10 and 3 mg/kg, respectively. These results indicate that rolipram ameliorates impairments of learning and memory in rats and mice, and suggest that rolipram might ameliorate the impairments of learning and memory by elevating cAMP levels.

Biphasic effects of D3-receptor agonists, 7-OH-DPAT and PD128907, on the D1-receptor agonist-induced hyperactivity in mice.

Effects of D3-receptor agonists, 7-OH-DPAT and PD128907, on the D1-receptor agonist SKF81297-induced hyperactivity in mice were examined. 7-OH-DPAT and PD128907 significantly suppressed the SKF81297-induced hyperactivity at low doses, but significantly potentiated the hyperactivity at high doses. These D3-agonists alone had no effect on the motor activity. A kappa-receptor agonist that reduces dopamine release had no effect on the SKF81297-induced hyperactivity. These results suggest that lower doses of 7-OH-DPAT and PD128907 may negatively influence the D1-receptor mediated behaviors via post synaptic D3-receptors. On the other hand, higher doses of these compounds may positively influence these behaviors via D2- or D3-receptors.

Amphetamine sensitization enhances regional c-fos expression produced by conditioned fear.

Chronically administered amphetamine can result in a paranoid psychosis that can be re-induced in former amphetamine abusers by psychological stressors. In an attempt to investigate the neurobiological correlates of this phenomenon, the present study examined the effects of prior D-amphetamine sensitization on regional c-fos expression induced by a psychological stressor. Rats received intermittent footshock in a distinctive environment for 30 min/day for three days. Three days after the last fear conditioning session, the animals received injections of saline or D-amphetamine (4 mg/kg, i.p.) once every second day for 16 days (eight injections in total). After a 14-day drug abstinent period, the animals were placed in the fear conditioning apparatus but without footshock. The amphetamine sensitization procedure significantly enhanced the effects of conditioned fear on c-fos expression in several brain regions. These included the cingulate cortex area 3, agranular insular cortex (layers 2 and 3), claustrum, piriform cortex, the shell region of the nucleus accumbens, medial striatum, ventral lateral septum, and CA3 and polymorphic layer of the hippocampal formation. These results indicate that D-amphetamine sensitization can have long-lasting effects on the neural circuitries activated by conditioned stressors.

5-HT1A-receptor subtype mediates the effect of fluvoxamine, a selective serotonin reuptake inhibitor, on marble-burying behavior in mice.

The effect of fluvoxamine, a selective serotonin (5-HT) reuptake inhibitor, was studied in a model of anxiety and/or obsessive compulsive disorder (OCD) in mice. In the anxiety/OCD model, marble-burying behavior, marble-burying was significantly suppressed by fluvoxamine at 30 and 60 mg/kg, p.o. and the monoamine reuptake inhibitor clomipramine, at 60 mg/kg, p.o. No suppressive effect, however, was observed by the selective norepinephrine reuptake inhibitor desipramine at doses from 15 to 60 mg/kg, p.o. Suppressive effects were obtained by the serotonergic anxiolytic buspirone at 30 and 60 mg/kg, p.o. and the benzodiazepine anxiolytic diazepam at 10 mg/kg, p.o. The effect of fluvoxamine on marble-burying was slightly attenuated after repeated administration. On the other hand, both the effects of buspirone and diazepam completely disappeared after repeated administration. Effect of fluvoxamine on the marble-burying was unaffected by the 5-HT2 antagonist ritanserin. However, the 5-HT1A antagonist NAN-190 (1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl] piperazine) inhibited the suppressive effect of fluvoxamine on the marble-burying. From these results, the 5-HT1A-receptor subtype may be involved in the suppressive effect of fluvoxamine on the marble-burying, but the 5-HT2-receptor subtype is not involved in this effect.

Neither the 5-HT1A- nor the 5-HT2-receptor subtype mediates the effect of fluvoxamine, a selective serotonin reuptake inhibitor, on forced-swimming-induced immobility in mice.

The effect of fluvoxamine, a selective serotonin (5-HT) reuptake inhibitor, was studied in the forced-swimming test, a model of depression, in mice. Fluvoxamine at 60 mg/kg, p.o. significantly decreased the immobility time in the forced-swimming test. A similar effect was observed by the selective norepinephrine reuptake inhibitor desipramine at the same dose. Furthermore, the suppression of immobility time was slightly potentiated by repeated administration of fluvoxamine, and a significant effect was observed at 30 mg/kg, p.o. The effect of fluvoxamine on forced-swimming was unaffected by the 5-HT2 antagonist ritanserin. On the other hand, the 5-HT1A antagonist NAN-190 (1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl] piperazine) potentiated the effect of fluvoxamine on forced-swimming. It is expected, however, that a 5-HT1A antagonist should antagonize the effect of fluvoxamine when 5-HT1A mediates the suppressive effect of fluvoxamine on the immobility time in forced-swimming. From these results, neither the 5-HT1A- nor the 5-HT2-receptor subtype is involved in the suppressive effect of fluvoxamine on the immobility associated with forced-swimming.

Computer analysis of Holter electrocardiogram.

Computer analysis is indispensable for the interpretation of Holter ECG, because it includes a large quantity of data. Computer analysis of Holter ECG is similar to that of conventional ECG, however, in computer analysis of Holter ECG, there are some difficulties such as many noise, limited analyzing time and voluminous data. The main topics in computer analysis of Holter ECG will be arrhythmias, ST-T changes, heart rate variability, QT interval, late potential and construction of database. Although many papers have been published on the computer analysis of Holter ECG, some of the papers was reviewed briefly in the present paper. We have studied on computer analysis of VPCs, ST-T changes, heart rate variability, QT interval and Cheyne-Stokes respiration during 24-hour ambulatory ECG monitoring. Further, we have studied on ambulatory palmar sweating for the evaluation of mental stress during a day. In future, the development of "the integrated Holter system", which enables the evaluation of ventricular vulnerability and modulating factor such as psychoneural hypersensitivity may be important.

[Klinefelter syndrome accompanied by hypoplasia of the bilateral internal carotid arteries--a case report].

The patient was a 31-year-old male who was admitted to our hospital due to slowly progressive weakness of the right upper limb and both lower limbs. Physical examination showed a tall stature, gynecomastia, feminine pubic hair, small penis and small testis. His intellectual function on the Kohs-cubic test scored 85, on the WAIS scored performance 80, verbal 74 and full scale 78. He showed BE mixed type in YG personality test and neurotic in CMI. No abnormalities were observed except for severe weakness of the right upper limb, moderate weakness of the bilateral lower limbs and hypalgesia in the limbs. Clinical laboratory tests revealed low blood and urinary testosterone levels, and a high blood gonadotropin level. Cytogenetic studies showed a 47, XXY karyotype. On neuroradiological examinations, there were small bilateral carotid canals in CT scan and multiple long T2 lesions in MRI. Angiography showed hypoplasia of the bilateral internal carotid arteries and dilatation of the bilateral vertebral arteries. The results of activity measurements of affected limbs using actigraph indicated that a conversion type hysteria had resulted in weakness of the limbs. There is no report of Klinefelter syndrome accompanied by the hypoplasia of bilateral internal carotid arteries, and the condition is considered to be extremely rare.

Rhythms in the central nervous system and 1/f fluctuations of the heart rate.

We have developed a system to analyze heart rate variability (HRV) (power spectral array of the HRV) during 24 h ambulatory electrocardiographic monitoring. Several rhythms (circadian and several ultradian rhythms) were observed in the power spectral array of the heart rates and 1/f-like fluctuations in the log-log scaled heart rate power spectrum. The circadian change of the heart rate is closely related to the body temperature rhythm. The 90 min rhythm of HRV during sleep was suspected to be produced by the sleep cycle (REM/NREM) and the lower frequency peak of the HRV was coherent with oscillation in amplitude modulated respiration. These circadian and ultradian rhythm as assessed by heart rate variability exist both in normal subjects and in patients with autonomic failure. The power of the high frequency band decreases in subjects with autonomic failure. The power of low frequency components increases during periodic breathing or Cheyne-Stokes respiration. Log-log scaled analysis of the power spectrum of HRV disclosed that the slope of the HRV is markedly modulated by the range of the frequency applied for the least square regression line analysis. The increased power that might be produced by periodic breathing and decreased power in patients with autonomic failure might strongly modulate the slope of the log-log scaled HRV. It is concluded that the power spectral array of the HRV during 24 h period is useful in the detection of circadian and ultradian rhythm, and log-log scaled power spectra might be useful in the overall integration of the heart rate dynamics produced by the central nervous system. The several rhythm factors that might be produced by the central nervous system might modulate 1/f fluctuations of the HRV.

Identification and isolation of endogenous insect phenoloxidase inhibitors.

Inhibitors of phenoloxidase were identified in pupae of the housefly, Musca domestica L. The phenoloxidase inhibitors were purified from final instar pupae of the housefly by a combination of ammonium sulfate fractionation, ion-exchange chromatography, gel filtration and reverse-phase high performance liquid chromatography. The potent phenoloxidase inhibitors were heat-stable low molecular weight peptides with an inhibition constant of nM range. To the best of our knowledge, this is the first time that endogenous phenoloxidase inhibitors have been identified among the insects, and probably also among the invertebrates. It is likely that the inhibitors play a central role in regulating the action of active phenoloxidases and will also serve as important tools for understanding the structures and functions of phenoloxidases, as well as their role in insect metamorphosis.