Distribution and drift dispersal dynamics of a caddisfly grazer in response to resource abundance and its ontogeny.

Stream grazers have a major impact on food web structure and the productivity of stream ecosystems; however, studies on the longitudinal (upstream versus downstream) and temporal changes in their drift dynamics and resulting distributions remain limited. Here, we investigated the longitudinal and temporal distributions and drift propensity of a trichopteran grazer, the caddisfly, Micrasema quadriloba, during its life cycle in a Japanese stream. The distribution of larvae significantly shifted downstream during the fifth instar larval stage during late winter; with periphyton abundance (i.e. their food source) showing similar shifts downstream. Therefore, our results show that the drift dispersal the caddisfly occurs in response to decline in available food resources (i.e. food-resource scarcity) and an increase in food requirements by growing individuals. Furthermore, our results show that this observed longitudinal shift in larval distribution varies through their life cycle, because the drift dispersal of fifth instar larvae was greater than that of immature larvae. The correlation between periphyton abundance and drift propensity of fourth instar larvae was not statistically significant, whereas that of fifth instar larvae was significantly negative. In conclusion, we detected an ontogenetic shift in drift propensity, which might explain the longitudinal and temporal distributions of this species.

The Parapineal Is Incorporated into the Habenula during Ontogenesis in the Medaka Fish.

The parapineal is present in many teleost families, while it is absent in several others. To find out why the parapineal is absent at adult stages in the latter families, the development of the epithalamus was examined in the medaka fish (Oryzias latipes). For this purpose, a green fluorescent protein-transgenic medaka line, in which the pineal complex (pineal and parapineal) is visible fluorescently, was used. We found that a distinct parapineal was present in the roof plate at early developmental stages. Subsequently, however, the parapineal and the associated roof plate began to be incorporated into the habenula between embryonic stages 28 and 29. Between embryonic stages 29 and 30, the entire parapineal was incorporated into the habenula. That is, the parapineal became a small caudomedial region (termed the 'parapineal domain') within the left habenula in the majority of embryos, resulting in the left-sided asymmetry of the epithalamus. Thereby the left habenula became larger and more complex than its right counterpart. In the minority of embryos, the parapineal was incorporated into the right habenula or into the habenulae on both sides. In the majority of embryos, the parapineal domain projected a fiber bundle to a subnucleus (termed the 'rostromedial subnucleus') in the left habenula. The rostromedial subnucleus sent axons, through the left fasciculus retroflexus, to the rostral region of the left half of the interpeduncular nucleus. We further found that the ratio of the left-sided phenotype was temperature dependent and decreased in embryos raised at a high temperature. The present study is the first demonstration that the supposed lack of a distinct parapineal in adult teleost fishes is due to ontogenetic incorporation into the habenula.

ß-arrestin functionally regulates the non-bleaching pigment parapinopsin in lamprey pineal.

The light response of vertebrate visual cells is achieved by light-sensing proteins such as opsin-based pigments as well as signal transduction proteins, including visual arrestin. Previous studies have indicated that the pineal pigment parapinopsin has evolutionally and physiologically important characteristics. Parapinopsin is phylogenetically related to vertebrate visual pigments. However, unlike the photoproduct of the visual pigment rhodopsin, which is unstable, dissociating from its chromophore and bleaching, the parapinopsin photoproduct is stable and does not release its chromophore. Here, we investigated arrestin, which regulates parapinopsin signaling, in the lamprey pineal organ, where parapinopsin and rhodopsin are localized to distinct photoreceptor cells. We found that beta-arrestin, which binds to stimulated G protein-coupled receptors (GPCRs) other than opsin-based pigments, was localized to parapinopsin-containing cells. This result stands in contrast to the localization of visual arrestin in rhodopsin-containing cells. Beta-arrestin bound to cultured cell membranes containing parapinopsin light-dependently and translocated to the outer segments of pineal parapinopsin-containing cells, suggesting that beta-arrestin binds to parapinopsin to arrest parapinopsin signaling. Interestingly, beta-arrestin colocalized with parapinopsin in the granules of the parapinopsin-expressing cell bodies under light illumination. Because beta-arrestin, which is a mediator of clathrin-mediated GPCR internalization, also served as a mediator of parapinopsin internalization in cultured cells, these results suggest that the granules were generated light-dependently by beta-arrestin-mediated internalization of parapinopsins from the outer segments. Therefore, our findings imply that beta-arrestin-mediated internalization is responsible for eliminating the stable photoproduct and restoring cell conditions to the original dark state. Taken together with a previous finding that the bleaching pigment evolved from a non-bleaching pigment, vertebrate visual arrestin may have evolved from a "beta-like" arrestin by losing its clathrin-binding domain and its function as an internalization mediator. Such changes would have followed the evolution of vertebrate visual pigments, which generate unstable photoproducts that independently decay by chromophore dissociation.

Influences of twilight on diurnal variation of core temperature, its nadir, and urinary 6-hydroxymelatonin sulfate during nocturnal sleep and morning drowsiness.

This study aimed at elucidating the physiological significance of dusk and dawn in the circadian rhythm of core temperature (T(core)) and urinary 6-hydroxymelatonin sulfate in humans during sleep and the waking sensation just after rising. Seven female and four male students served as participants. Participants retired at 2300 h and rose at 0700 h. They were requested to sit on a chair and spend time as quietly as possible during wakefulness, reading a book or listening to recorded light music. Two lighting conditions were provided for each participant: 1) Light-Dark (LD)-rectangular light change with abrupt decrease from 3,000 lx to 100 lx at 1800 h, abrupt increase from 0 lx to 3,000 lx at 0700 h. 2) LD-twilight light change with gradual decrease from 3,000 lx to 100 lx starting at 1700 h (twilight period about 2 h), with gradual increase from 0 lx to 3,000 lx starting at 0500 h (twilight period about 2 h). The periods of 0 lx at night were from 2300 h to 0700 h on the first day and from 2300 to 0500 h on the second day. Nadir time advanced significantly under the influence of the LD-twilight condition. The amount of 6-hydroxymelatonin sulfate in urine collected at 0200 h was significantly higher under LD-twilight in comparison with LD-rectangular light. Morning drowsiness tended to be lower under LD-twilight. Our results suggest that in architectural design of indoor illumination it is important to provide LD-twilight in the evening and early morning for sleep promotion in healthy normal people and/or light treatment in elderly patients with advanced dementia.

Immunohistochemical characterization of a parapinopsin-containing photoreceptor cell involved in the ultraviolet/green discrimination in the pineal organ of the river lamprey Lethenteron japonicum.

In the pineal organ, two types of ganglion cell exhibit antagonistic chromatic responses to UV and green light, and achromatic responses to visible light. In this study, we histologically characterized UV-sensitive photoreceptor cells that contain a unique non-visual UV pigment, lamprey parapinopsin, in order to elucidate the neural network that is associated with antagonistic chromatic responses. These characteristics were compared with those of lamprey rhodopsin-containing cells, most of which are involved in achromatic responses. RT-PCR analysis revealed that lamprey parapinopsin was expressed in the pineal organ but not in the retina, unlike lamprey rhodopsin, which was expressed in both. Lamprey parapinopsin and lamprey rhodopsin were immunohistochemically localized in the dorsal and ventral regions of the pineal organ, respectively. The two pigments were localized in distinct photoreceptor cells throughout the pineal organ, namely the dorsal and ventral regions as well as the peripheral region, which corresponds to the dorso-ventral border region. The ratio of the number of lamprey parapinopsin-containing cells to lamprey rhodopsin-containing cells around the peripheral region was higher than in the central region. Electron-microscopic analysis revealed that lamprey parapinopsin-containing dorsal cells have outer segments and synaptic ribbons similar to those of ventral photoreceptor cells. However, unlike lamprey rhodopsin-containing cells, lamprey parapinopsin-containing cells connected with each other in a wide area of dorsal and peripheral portions and made direct contact with ganglion cells, mainly in the peripheral portion. These results suggest that UV light information captured by lamprey parapinopsin-containing photoreceptor cells is converged and directly transmitted to chromatic-type ganglion cells in the peripheral region to generate antagonistic chromatic responses.

Combined influences of gradual changes in room temperature and light around dusk and dawn on circadian rhythms of core temperature, urinary 6-hydroxymelatonin sulfate and waking sensation just after rising.

The present experiment aimed at knowing how a gradual changes of room temperature (T(a)) and light in the evening and early morning could influence circadian rhythms of core temperature (T(core)), skin temperatures, urinary 6-hydroxymelatonin sulfate and waking sensation just after rising in humans. Two kinds of room environment were provided for each participant: 1) Constant room temperature (T(a)) of 27 degrees C over the 24 h and LD-rectangular light change with abrupt decreasing from 3,000 lx to 100 lx at 1800, abrupt increasing from 0 lx to 3,000 lx at 0700. 2) Cyclic changes of T(a) and with gradual decrease from 3,000 lx to 100 lx onset at 1700 (twilight period about 2 h), with gradual increasing from 0 lx to 3,000 lx onset at 0500 (about 2 h). Main results are summarized as follows: 1) Circadian rhythms of nadir in the core temperature (T(core)) significantly advanced earlier under the influence of gradual changes of T(a) and light than no gradual changes of T(a) and light. 2) Nocturnal fall of T(core) and morning rise of T(core) were greater and quicker, respectively, under the influence of gradual changes of T(a) and light than no gradual changes of T(a) and light. 3) Urinary 6-hydroxymelatonin sulfate during nocturnal sleep was significantly greater under the influence of gradual changes of T(a) and light. 4) Waking sensation just after rising was significantly better under the influence of gradual changes of T(a) and light. We discussed these findings in terms of circadian and thermoregulatory physiology.

Physiological significance of cyclic changes in room temperature around dusk and dawn for circadian rhythms of core and skin temperature, urinary 6-hydroxymelatonin sulfate, and waking sensation just after rising.

The present study investigated whether room temperature (T(a)) cycles around dusk and dawn could influence the circadian rhythm of rectal temperature (T(core)), urinary 6-hydroxymelatonin sulfate during nocturnal sleep, and subjective assessments of sleep in humans. Six female and two male students served as participants. Two different T(a) conditions, cyclic and constant, were established. Two kinds of room temperature were provided to subjects: cyclic T(a) (gradual decrease from 27 degrees C to 24 degrees C between 1800 and 2200 h and gradual increase from 24 degrees C to 27 degrees C between 0300 and 0700 h) and constant T(a) (27 degrees C over 24 h). At cyclic T(a), the circadian nadir of T(core) rhythm was significantly advanced, while T(core) was significantly lower from 2300 to 0200 h and significantly higher from 0600 to 1000 h. The nocturnal concentration of 6-hydroxymelatonin sulfate in the urine during sleep was significantly higher during cyclic T(a). Waking sensation just after rising was significantly better with cyclic T(a). (Skin temperatures in the extremities-T(a)) were significantly higher with cyclic T(a) especially during the evening and night. Our results suggest that gradual change of room temperature in the evening and early morning is important in terms of sleep promotion and fresh awakening. It seems probable that mankind has been evolved to have deeper sleep under the influence of cyclic T(a) around dusk and dawn.

Neural interaction of gonadotropin-regulating hormone immunoreactive neurons and the suprachiasmatic nucleus with the paraventricular organ in the Japanese grass lizard (Takydromus tachydromoides).

Our previous study demonstrated that the paraventricular organ (PVO) in the hypothalamus of the Japanese grass lizard (Takydromus tachydromoides) showed immunoreactivity against the light signal-transducing G-protein, transducin. This finding suggested that the PVO was a candidate for the deep-brain photoreceptor in this species. To understand functions of the PVO, we investigated distributions of transducin, serotonin, gonadotropin-releasing hormone (GnRH), and gonadotropin-inhibitory hormone (GnIH) in the lizard's brain. We immunohistochemically confirmed co-localization of transducin and serotonin in PVO neurons that showed structural characteristics of cerebrospinal fluid (CSF)-contacting neurons. GnRH-immunoreactive (ir) cells were localized in the posterior commissure and lateral hypothalamic area. Some of the serotonin-ir fibers extending from the PVO to the lateral hypothalamic area contacted the GnRH-ir cell bodies. GnIH-ir cells were localized in the nucleus accumbens, paraventricular nucleus, and upper medulla, and GnIH-ir fibers from the paraventricular nucleus contacted the lateral processes of serotonin-ir neurons in the PVO. In addition, we found that serotonin-ir fibers from the PVO extended to the suprachiasmatic nucleus (SCN), and the retrograde transport method confirmed the PVO projections to the SCN. These findings suggest that the PVO, by means of innervation mediated by serotonin, plays an important role in the regulation of pituitary function and the biological clock in the Japanese grass lizard.

Histological changes in immune and endocrine organs of quail embryos: exposure to estrogen and nonylphenol.

Effects of 17beta-estradiol and p-nonylphenol were examined in the immune and endocrine organs of Japanese quail embryos. The test compounds were injected into the yolk of embryonated eggs. Injection of estrogen resulted in (1) disappearance of lymphoid cells and flattened development of plicae in the bursa of Fabricius, (2) decreased area of thyroid follicles and height of simple cuboidal epithelial cells in the thyroid, (3) increased follicular appearance of the thymus, and (4) development of an ovotestis in male embryos. Injection of nonylphenol did not induce flattened plicae in the bursa of Fabricius or development of an ovotestis in male embryos, but it increased the disappearance of lymphoid cells from the lymphoid follicles in the bursa, decreased the height of simple cuboidal epithelial cells surrounding the thyroid follicle, and increased the follicle-like structure in the thymus in male embryos. These results suggest that nonylphenol has estrogenic effects, but these are low compared to those of estrogen itself.

Reach-scale distribution dynamics of a grazing stream insect, Micrasema quadriloba Martynov (Brachycentridae, Trichoptera), in relation to current velocity and periphyton abundance.

Reach-scale temporal shifts in the distribution of larvae of a grazing caddisfly, Micrasema quadriloba (Brachycentridae), were investigated in a Japanese mountain stream. The larvae showed an aggregated distribution within the reach at the beginning of the immigration, then became randomly dispersed throughout the reach as the immigration progressed. The abundance of periphyton in the reach decreased dramatically with increasing dispersal of the larvae. Simple regression analyses revealed that the stream's flow regime was the most important environmental factor that determined the reach-scale distribution of the larvae and that the relationship between the flow regime and the distribution of the larvae shifted temporally. In addition, our results suggest that only this species of grazing insect, which was dominant in the study reach, controlled the reach-scale abundance of the periphyton.

Effects of estrogen (17 beta-estradiol) and p-nonylphenol on the development of immune organs in male Japanese quail.

The aim of this study was to determine the effects of estrogen and endocrine disrupters on the immune organs of Japanese quail. We used p-nonylphenol (NP) as a model endocrine disrupter. Birds were injected with 17 beta-estradiol (E2) in doses of 1, 0.1 and 0.01 ng/g body weight and NP in doses of 1000, 100 and 10 ng/g body weight from 4 to 7 weeks of age. The data indicate injection of E2 during the growth phase resulted in cyst formation, increased connective tissue, disappearance of lymphoid cells and disappearance of plicae in the bursa, disappearance of lymphocytes in the thymus and a greater population of trabecular arteries and veins in the spleen. Injection of NP tended to induce many empty vacuoles and increased connective tissue in the bursa of Fabricius but did not affect the structures of the spleen and thymus. We also studied whether apoptosis is involved in the histological changes of the bursa and found that apoptosis did not play an important role in the changes of the bursa. These results suggest that E2 and NP may affect the immune organs and the effect on the lymphoid tissue in immune organs is higher with E2 treatment than with NP.

Vitellogenin-immunohistochemistry in the liver and the testis of the medaka, Oryzias latipes, exposed to 17beta-estradiol and p-nonylphenol.

Vitellogenin (VTG) produced in male fish has been used for a biomarker to study endocrine disrupters. However, the characteristics of VTG produced in male fish have not been studied well. In this study, we investigated the localization of VTG in the liver and the testis of male medaka (Oryzias latipes) treated with 17beta-estradiol (E2) and p-nonylphenol (NP). The male fish were exposed to 1 microg/L E2 and 500 microg/L NP for 1-12 days. Control groups were kept in water including only vehicle. The frozen sections of the liver and the testis were stained with immunohistochemical methods using an antiserum against medaka VTG as the first antibody. In the E2 and NP treated liver, the hepatocytes showed immunoreactivity. In particular, the cytoplasm close to the cell membrane surrounding the sinusoids was strongly immunopositive. In the testis of both treatments, the interstitial tissues and the cells (spermatocytes) in the seminiferous tubules were immunopositive. The concentration of VTG became gradually higher in both tissues with longer treatments. These results suggest that germ cells in the testis treated with E2 and NP are able to incorporate and accumulate VTG.

Association between moringness-eveningness preference and mental/physical premenstrual symptoms in Japanese females 12 to 31 years of age.

This study investigates the relationship between circadian typology, i.e., morningness-eveningness (M-E) preference, and the occurrence and severity of premenstrual mental and physical symptoms among 154 young Japanese female university students (range, 18 to 31 yrs; mean+/-S.D., 20.69+/-3.69 yrs) and 417 junior high school students (range, 12 to 15 yrs; mean+/-S.D., 14.29+/-0.67 yrs) living in an urban or suburban area of Kochi prefecture. Female university students experienced melancholy mood more frequently than did males, and the female university students who frequently became melancholy were more evening-typed than those who did not experience melancholy. Female university students who experienced frequent fluctuations in mood and/or menstrual pain were more evening-typed than those who were not so affected. M-E preference of junior high school students was not correlated with stability of mood or frequency of menstrual pain. In urban areas, however junior high school students who had very stable menstrual cycles were significantly more morning-typed than those whose menstrual cycles were not stable. In suburban areas, the bedtimes of female junior high students who had stable menstrual cycles were significantly earlier than those whose menstrual cycle duration was not stable. A physiological relationship between the circadian system, M-E, and attributes of the menstrual cycle seems to be present in adolescent female Japanese junior high school students.

Effect of dim and bright light exposure on some immunological parameters measured under thermal neutral conditions.

This study assesses the effects of ambient light conditions, under a thermoneutral environment, on selected immunological parameters of 7 healthy young women (aged 19 to 22 yrs). Subjects entered the bioclimatic chamber at 11: 00 h, controlled at 26 degrees C and 60% relative humidity, a "neutral climate". They lead a well-regulated life in the climatic chamber (pre-condition) while exposed to dim (200 lux) or, on the next day, bright (5000 lux) light between 06 : 00 to 12 : 00 h. Just before the end of each period of light exposure, a blood sample was taken for later immunological assay of white blood cell count (WBC), phagocytosis, interferon-gamma (IFN-gamma), interleukin-4 (IL-4), CD69 T cells (CD69), CD4+CD25+ T cells (CD4+CD25+), and transforming growth factor-beta 1 (TGF-beta1). The results, when compared with the pre-condition, were as follows: 1) CD69 and IFN-gamma increased during normal conditions without thermal stress under dim light; 2) WBC increased and IL-4 decreased under bright light; 3) as shown by the highly significant decrease of TGF-beta1, the immune system was activated under bright light; 4) phagocytosis tended to increase under bright light exposure; 5) CD69 and IFN-gamma were significantly higher, and CD4+CD25+ tended to decrease under bright light; 6) phagocytosis tended to be lower and TGF-beta1 significantly higher under dim light, indicating a decline of immune system function. Taken together, this preliminary single time-point sampling study infers that some parameters are activated (CD69) while others are attenuated (phagocytosis, TGF-beta1) according to the environmental light intensity, dim vs. bright, in women adhering to a standardized routine in the absence of thermal stress. These findings are discussed in terms of inhibition of the sympathetic and excitation of the parasympathetic nervous system under the influence of life-style regularity and daytime bright light exposure.

Photoperiodic response of serotonin- and galanin-immunoreactive neurons of the paraventricular organ and infundibular nucleus in Japanese quail, Coturnix coturnix japonica.

We investigated the photoperiodic response of serotonin- and galanin (GA)- immunoreactive (ir) cells in the paraventricular organ (PVO) and infundibular nucleus (IF) of the Japanese quail and the interaction of these cells with gonadotropin-releasing hormone (GnRH)-ir neurons in the hypothalamus. Serotonin-ir cells were located in series from the PVO to the IF, and were connected with each other. The number of serotonin-ir cells differed significantly between light and dark phases on the short days (SD), but did not differ between light and dark phases on long days (LD). GA-ir cells were also found in the PVO and IF. The number of GA-ir cells under SD conditions was significantly greater than under LD conditions but did not change diurnally. Both serotonin-ir and GA-ir fibers ran along the GnRH-ir cells in the nucleus commissurae pallii. Serotonin-ir and GA-ir fibers were connected with the GnRH-ir fibers in the external layer of the median eminence (ME). We confirmed that GA-ir fibers were closely associated with serotonin-ir neurons in the PVO and IF. GA-ir neurons have at least 2 routes of regulating GnRH neurons directly, and indirectly via the serotonin-ir cells in the PVO and IF.

Bistable UV pigment in the lamprey pineal.

Lower vertebrates can detect UV light with the pineal complex independently of eyes. Electrophysiological studies, together with chromophore extraction analysis, have suggested that the underlying pigment in the lamprey pineal exhibits a bistable nature, that is, reversible photoreaction by UV and visible light, which is never achieved by known UV pigments. Here we addressed the molecular identification of the pineal UV receptor. Our results showed that the long-hypothesized pigment is a lamprey homologue of parapinopsin, which exhibits an absorption maximum at 370 nm, in the UV region. UV light causes cis-trans isomerization of its retinal(2) chromophore, forming a stable photoproduct having an absorption maximum at 515 nm, in the green region. The photoproduct reverts to the original pigment upon visible light absorption, showing photoregeneration of the pigment. In situ hybridization showed that parapinopsin is selectively expressed in the cells located in the dorsal region of the pineal organ. We successfully obtained the hyperpolarizing responses with a maximum sensitivity of approximately 380 nm from the photoreceptor cells at the dorsal region, in which the outer segment was clearly stained with anti-parapinopsin antibody. These results demonstrated that parapinopsin is the pineal UV pigment having photointerconvertible two stable states. The bistable nature of the parapinopsin can account for the photorecovery of the pineal UV sensitivity by background green light in the lamprey. Furthermore, we isolated the parapinopsin homologues from fish and frog pineal complexes that exhibit UV sensitivity, suggesting that parapinopsin is a common molecular basis for pineal UV reception in the vertebrate.

Variations in plasma melatonin levels of the rainbow trout (Oncorhynchus mykiss) under various light and temperature conditions.

Daily variations in plasma melatonin levels in the rainbow trout Oncorhynchus mykiss were studied under various light and temperature conditions. Plasma melatonin levels were higher at mid-dark than those at mid-light under light-dark (LD) cycles. An acute exposure to darkness (2 hr) during the light phase significantly elevated the plasma melatonin to the level that is comparable with those at mid-dark, while an acute exposure to a light pulse (2 hr) during the dark phase significantly suppressed melatonin to the level that is comparable with those at mid-light. Plasma melatonin kept constantly high and low levels under constant darkness and constant light, respectively. No circadian rhythm was seen under both conditions. When the fish were subjected to simulative seasonal conditions (simulative (S)-spring: under LD 13.1:10.9 at 13 degrees C; S-summer: under LD 14.3:9.7 at 16.5 degrees C; S-autumn: under LD 11.3:12.7 at 13 degrees C; S-winter: under LD 10.1:13.9 at 9 degrees C), melatonin levels during the dark phase were significantly higher than those during the light phase irrespective of simulative seasons. The peak melatonin level in each simulative season significantly correlated with temperature but not with the length of the dark phase employed. In addition, the peak melatonin level in S-autumn was significantly higher than those in S-spring although water temperature was the same under these conditions. These results indicate that the melatonin rhythm in the trout plasma is not regulated by an endogenous circadian clock but by combination of photoperiod and water temperature.

The relation among emergence date, days until first effective mating, fecundity and adult longevity in Isoperla aizuana and Sweltsa sp. (Plecoptera).

The relation between emergence date and the factors concerned in reproduction were studied in the stoneflies Isoperla aizuana (Perlodidae) and Sweltsa sp. (Chloroperlidae) in the laboratory. Individuals emerging earlier had larger adult size and greater adult longevity. Fecundity was also related to emergence date, adult size, and adult longevity. Furthermore, individuals emerging earlier had more days until first effective mating and tended to mate more effectively than later-emerging individuals. The maturity of the reproductive tissue may differ with emergence date. While most of the later-emerging individuals had a lower rate of effective mating, longer-lived and larger size adults mated more effectively in females of Sweltsa sp. The differences in mating character might affect the effectiveness of mating.

Thermal comfort zones in the Vietnamese.

Thermally comfortable zones in Vietnamese were investigated during winter in Hanoi. The subjects were 21 males (age: 19.7 +/- 0.4 yrs; height: 165 +/- 1.5 cm; body mass: 55.1 +/- 1.1 kg) and 19 females (age: 19.7 +/- 0.4 yrs; height; 155.6 +/- 1.7 cm; body mass: 45.6 +/- 1.3 kg). Each participant entered singly the climatic chamber controlled at 22 degrees C and 40% RH. After 20 min rest, the participant was requested to indicate on a 7-point scale (Table 1) how he or she felt to the room temperature given. Then, the room temperature increased by 1 degrees C over 10 min every 20 min. Just before the rise of the room temperature, the participant judged his or her thermal sensation. More than 90% of the participants felt 24-29 degrees C of the room temperature as "slightly cool", "neutral" and "slightly warm" (Table 2). We defined these sensations as "thermally comfort". These thermally comfortable zones were quite higher than those (20-24 degrees C) recommended by ISO-7730 (1994). We discussed these discrepancies in terms of higher establishment of thermoregulatory set-point in the Vietnamese.

Seasonal changes in neurosteroid concentrations in the amphibian brain and environmental factors regulating their changes.

Up to now the regulatory mechanisms, which govern the concentrations of neurosteroids in the brain, are unclear. Seasonal breeders may serve as excellent models to understand physiological changes in neurosteroid levels and their regulatory mechanisms. The present study first investigated immunohistochemically the localization of cytochrome p450 side-chain cleavage enzyme (p450scc) and 3beta-hydroxysteroid dehydrogenase/delta(5)-delta(4)-isomerase (3betaHSD) in the brain of the newt Cynops pyrrhogaster, a seasonally breeding amphibian. Both p450scc- and 3betaHSD-like immunoreactive cells were restricted to the preoptic area. Seasonal changes in neurosteroid concentrations were then examined using adult males. Pregnenolone concentrations in the brain showed marked changes during annual breeding cycle and a maximal level in August, independent of the plasma steroid levels which were all low throughout the year. Progesterone concentrations in the brain, which were lower than pregnenolone levels, also showed peaks in April and August. In contrast, the pregnenolone sulfate level was low and its change was less pronounced. To clarify environmental factors that induce seasonal changes in neurosteroid levels, adult males were further subjected to different photoperiods and ambient temperatures for 5 weeks. Both pregnenolone and progesterone concentrations in the brain were significantly higher in the long day (LD) group than in the short day (SD) group, whereas no significant effects of different ambient temperatures on neurosteroid levels were detected. These results suggest that the newt brain possesses steroidogenic enzymes p450scc and 3betaHSD and exhibits seasonal changes in the pregnenolone and progesterone concentrations. Photoperiod may be a more important environmental factor than temperature for the regulation of neurosteroid levels in the brain.