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M Iigo

Publications and source records attributed to M Iigo.

At least 37 records · Page 2Linked to original sources

Light-dependent expression of pinopsin gene in chicken pineal gland.

The phase of a circadian clock in the chicken pineal gland is reset by an environmental light signal, which is captured by the pineal photoreceptive molecule(s). Here we show that the mRNA level of pinopsin, a predominant photoreceptive molecule in the pineal gland, undergoes a diurnal fluctuation in chickens maintained on a light/dark cycle. The mRNA levels in the light were approximately six times higher than those in the dark. This fluctuation was not observed in constant darkness, where the mRNA levels remained low. Subsequent light exposure of chickens increased the amount of pinopsin mRNA regardless of the circadian time. Clearly, the expression of pinopsin gene is controlled by a light signal, independent of the circadian clock. In vitro experiments using cultured pineal glands isolated from the visual system also revealed the light-dependent increase in pinopsin mRNA level, indicating that the pineal photoreceptive molecule(s) is responsible for the induction. These results demonstrate the presence of a feedback loop through which the light signal captured by pinopsin stimulates the transcription of its own gene in the chicken pineal gland. In contrast, pinopsin protein remained at an almost constant level in chickens maintained under the same light/dark cycles. The protein level, however, decreased to approximately 50% of the light/dark level under constant darkness and subsequently increased upon exposure to light after the dark period. It is suggested that, under the light/dark cycles, the pinopsin protein level is kept constant by the light-dependent synthesis, which would compensate for a possible degradation of pinopsin in the daytime.

Animals↗

Inhibition of initiation and early stage development of aberrant crypt foci and enhanced natural killer activity in male rats administered bovine lactoferrin concomitantly with azoxymethane.

The influence of concomitant administration of bovine lactoferrin (bLF) on induction of aberrant crypt foci (ACF) by azoxymethane was investigated in male F344 rats. Two percent bLF and 3% Bifidobacterium longum (B. longum), as a positive control, significantly decreased the numbers of ACF as well as the total numbers of aberrant crypts reproducibly in three independent studies (2% bLF, P < 0.01; 3% B. longum, P < 0.05). Most importantly large size foci composed of four or more crypts were always significantly decreased by 2% bLF (P < 0.05). Additional investigation of the natural killer activity of spleen cells demonstrated enhancement by bLF (P < 0.01) and B. longum (P < 0.01) in line with the levels of influence on foci induction, indicating a possible role for elevated immune cytotoxicity in the observed inhibition.

Animals↗

Inhibitory effects of oleic and docosahexaenoic acids on lung metastasis by colon-carcinoma-26 cells are associated with reduced matrix metalloproteinase-2 and -9 activities.

In order to determine the effects of single unsaturated fatty acids (UFAs) or combinations on establishment of lung metastatic colonies, UFAs were administered orally to CDF1 mice bearing s.c. implants of the highly metastatic colon carcinoma 26. Oleic acid (OA), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) demonstrated significant inhibition. In the case of DHA, this inhibitory potential was markedly reduced by co-administration of linoleic acid (LA) or EPA. Furthermore, while tumor cells treated with DHA showed a very low potential for lung colony formation when injected i.v., this again being partially reversed by co-administration of EPA. UFAs were found to be well absorbed into tumor tissues after oral administration, causing marked changes in relative levels, the arachidonic acid (AA) content, in particular, being markedly decreased by treatment with DHA or EPA, but not with DHA plus EPA or with DHA plus LA. Investigation of the gelatinolytic activity of the 57-kDa and 92-kDa isoforms of type-IV collagenase (MMP-2 and MMP-9, respectively) showed a clear reduction in the former by treatment with OA, while DHA, but not DHA plus LA or EPA, caused a decrease in the 92-kDa isoform, which was well correlated with AA content in tumor tissues (r = 0.900, p < 0.001). These results suggest that inhibition of metastasis due to treatment with OA and DHA might be due to depressed type-IV collagenase activity.

Animals↗

Unusual responses to light and darkness of ocular melatonin in European sea bass.

Regulation by light and darkness of melatonin rhythms in the plasma and eye of the European sea bass (Dicentrarchus labrax) was studied. During light-dark cycles, plasma and ocular melatonin exhibited day-night changes with higher levels at mid-dark and at mid-light, respectively. Circulating melatonin levels were low in constant light but high in constant darkness (DD); ocular melatonin levels showed the reverse pattern. Plasma melatonin exhibited circadian rhythm for 1 cycle but the rhythm was no longer apparent on day 2. There was no circadian rhythm in ocular melatonin. Acute light exposure in DD decreased plasma melatonin but increased ocular melatonin. These results suggest that circulating melatonin may be used as a signal for darkness but ocular melatonin is used as a signal for the light phase.

Animals↗

Characterization, guanosine 5'-O-(3-thiotriphosphate) modulation, daily variation, and localization of melatonin-binding sites in the catfish (Silurus asotus) brain.

Characteristics, guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) modulation, daily variation, and localization of melatonin-binding sites in the brain of a nocturnal teleost, the catfish Silurus asotus, were studied by radioreceptor assay using 2-[125I]iodomelatonin as the radioligand. The specific binding was rapid, stable, saturable, and reversible. The radioligand binds to a single class of receptor site with an affinity (Kd) of 30.7 +/- 7.3 pM and total binding capacity (Bmax) of 9.76 +/- 0.79 fmol/mg protein (mean +/- SE, n = 5). The binding sites were highly specific for 2-iodomelatonin and melatonin. The specificity was almost identical to that of functional melatonin receptors in the dermal and epidermal melanophores in this species and that of ML-1 subtype melatonin receptors in vertebrates, including melatonin-binding sites in the goldfish brain. GTPgammaS treatment altered both the Kd and Bmax values, indicating that melatonin-binding sites in the catfish brain are coupled to G protein. The Bmax values exhibited no daily variation under light-dark cycles of 12 hr light:12 hr dark whereas plasma melatonin levels and Kd fluctuated in a rhythmic fashion. The density of melatonin-binding sites in discrete brain areas was determined to be highest in optic tectum-thalamus and hypothalamus, intermediate in telencephalon, cerebellum, and medulla oblongata, and lowest in olfactory bulbs. These results suggest that melatonin secreted from the pineal organ and/or retina plays neuromodulatory roles in the catfish brain via G protein-coupled melatonin receptors. Characteristics of melatonin receptors seem to be highly conserved during evolution, although the density of melatonin receptors is not regulated by melatonin itself in this species.

Animals↗

Markedly induced asialoGM1+CD8+ T cell production and enhancement of antimetastatic activity by interferon beta with folic or folinic acid.

Either folic or folinic acid enhanced the antimetastatic activity of recombinant murine interferon beta (rMulFN beta) toward highly metastatic colon carcinoma 26 (Co 26Lu). Folinic acid administered with rMuIFN beta markedly increased asialoGM1+CD4+ and asialoGM1+CD8+ T cell production in the peritoneal cavity but not in the thymus and spleen. Peritoneal cells expressed killing activity toward Co 26Lu cells in vitro. In athymic nude mice, the above combination produced many asialoGM1+CD4+ and few asialoGM1+CD8+ T cells in the peritoneal cavity, but did not decrease lung metastatic colonies. AsialoGM1+CD4+ T cells would thus appear to have no or only very weak killing activity toward these tumor cells. The antimetastatic activity of folinic acid with rMuIFN beta was significantly decreased with anti-asialoGM1 and anti-CD8 antibodies. Inactivated CD8+ and asialoGM1+ cells cease to have killing activity toward Co 26Lu cells as shown by Winn's assay. AsialoGM1+CD8+ cell production was markedly induced in the peritoneal cavity by treatment with rMuIFN beta and folinic acid. AsialoGM1+CD8+ T cells may be inhibiting lung metastasis of Co 26Lu. Folinic acid and interferon are used in combination therapy with 5-fluorouracil for biochemical modulation. Folinic acid with interferon, as adjuvant therapy, may promote the induction of CD8+ T cell production with consequent prevention of metastasis.

Animals↗

Inhibitory effects of docosahexaenoic acid on colon carcinoma 26 metastasis to the lung.

Unsaturated fatty acids, including n-3 polyunsaturated fatty acids (PUFAs) such as docosahexaenoic acid (C22:6, DHA) and eicosapentaenoic acid (C20:5, EPA), and a series of n-6 PUFAs were investigated for their anti-tumour and antimetastatic effects in a subcutaneous (s.c.) implanted highly metastatic colon carcinoma 26 (Co 26Lu) model. EPA and DHA exerted significant inhibitory effects on tumour growth at the implantation site and significantly decreased the numbers of lung metastatic nodules. Oleic acid also significantly inhibited lung metastatic nodules. Treatment with arachidonic acid showed a tendency for reduction in colonization. However, treatment with high doses of fatty acids, especially linoleic acid, increased the numbers of lung metastatic nodules. DHA and EPA only inhibited lung colonizations when administered together with the tumour cells, suggesting that their incorporation is necessary for an influence to be exerted. Chromatography confirmed that contents of fatty acids in both tumour tissues and plasma were indeed affected by the treatments. Tumour cells pretreated with fatty acids in vivo, in particular DHA, also showed a low potential for lung colony formation when transferred to new hosts. Thus, DHA treatment exerted marked antimetastatic activity associated with pronounced change in the fatty acid component of tumour cells. The results indicate that uptake of DHA into tumour cells results in altered tumour cell membrane characteristics and a decreased ability to metastasize.

Animals↗

Chromosomal mapping of the gene encoding serotonin N-acetyltransferase to rat chromosome 10q32.3 and mouse chromosome 11E2.

Pineal melatonin is produced during the night. Its nocturnal increase regulates circadian rhythms and the photoperiodic reproductive response. Serotonin is acetylated to N-acetylserotonin by serotonin N-acetyltransferase (SNAT) and then methylated to form melatonin by hydroxyindole-O-methyltransferase (HIOMT). The rhythmicity of melatonin synthesis is regulated by the rhythmic activity of SNAT. Most laboratory mice do not have melatonin because of a genetic defect in the activity of SNAT and/or HIOMT. In a previous study using a recombinant inbred strain, we have found that the locus controlling pineal SNAT activity (Nat4) is located on mouse Chromosome 11. Recently, SNAT has been cloned in the rat. In the present study, the gene encoding SNAT was localized, using a rat cDNA fragment, on rat and mouse chromosomes by direct R-banding fluorescence in situ hybridization (FISH). In addition, using molecular linkage analysis with interspecific backcross mice, a gene encoding SNAT was mapped on a mouse chromosome. The gene encoding SNAT was localized to rat chromosome 10q32.3 and mouse Chromosome 11E2 by FISH. The molecular linkage analysis demonstrated that the gene encoding SNAT maps 1.5 cM distal to D11Mit11. The data suggest that Nat4 encodes SNAT. These chromosomal locations are in a region of conserved linkage homology between the two species.

Animals↗

Ocular melatonin rhythms in the goldfish, Carassius auratus.

Ocular melatonin rhythms in the goldfish were studied and compared to those in the pineal organ and plasma. Under light:dark (LD) of 12 h light:12 h dark, melatonin contents in the eye as well as the pineal organ and plasma exhibited clear day-night changes with higher levels at mid-dark than at mid-light. However, melatonin contents in the eye at mid-light and mid-dark were approximately 100 and 9 times greater than those in the pineal organ, respectively. Day-night changes of ocular melatonin persisted after pinealectomy, which abolished those in plasma melatonin under LD 12:12. Ocular melatonin contents in the pinealectomized fish at mid-light were significantly higher than those in the sham-operated control. Under constant darkness (DD), circadian melatonin rhythms were observed in the eye but damped on the 3rd day, whereas plasma melatonin rhythms generated by the pineal organ persisted for at least 3 days. Under constant light, ocular melatonin contents exhibited a significant fluctuation with a smaller amplitude than that under DD, whereas plasma melatonin remained at low levels. These results indicate the involvement of LD cycles, a circadian clock, and the pineal organ in the regulation of ocular melatonin rhythms in the goldfish.

Animals↗

Circadian rhythms of locomotor activity in the goldfish Carassius auratus.

Locomotor activities in the goldfish Carassius auratus were recorded under light-dark (LD) and constant light regimens. Under LD 12:12, all goldfish adjusted to given LD cycles. Activity patterns were classified into three types: L-type activity during the photophase (83%), LD-type activity both during the photophase and the scotophase (10%), and D-type activity during the scotophase (7%). However, these patterns were not fixed, but flexible. Under constant conditions, goldfish exhibited circadian locomotor activities: The proportions of appearance of circadian rhythms and tau values under constant darkness (DD), constant light (LL, 500 lx) and constant dim light (dimLL, 20 lx) were 57% (24.4 +/- 1.6 h, [mean +/- SE], n = 34), 57% (25.2 +/- 2.5 h, n = 35) and 67% (26.0 +/- 3.4 h, n = 10), respectively. These results indicate that the goldfish exhibit flexibility in phasing of locomotor activity rhythms under LD 12:12. In addition, the coupling between circadian clocks and locomotor activities does not seem strong, although the diel locomotor activity in the goldfish is regulated, in part, by a circadian clock. These results are discussed in context with the structure of the teleostean circadian system.

Animals↗

Demand feeding and locomotor circadian rhythms in the goldfish, Carassius auratus: dual and independent phasing.

In contrast to the common diurnal and nocturnal ways of life, some fish species have been shown to have a dual phasing behaviour. Therefore, the daily pattern of behaviour is not always rigidly confined to the light or dark phase and a diurnal fish may become nocturnal and vice versa. In the present study, the locomotor and feeding activities of single goldfish were simultaneously investigated to examine the existence of such dual behaviour. Nineteen goldfish weighing 97.2 g on average were placed individually in 35-1 glass tanks equipped with an infrared sensor and a newly developed self-feeding device. Fish were exposed to a light:dark (LD) 12:12 h cycle, constant darkness (DD), and 45:45 min LD pulses to study endogenous rhythmicity. Under LD 12:12, the daily pattern of behaviour differed between individual fish; some goldfish were diurnal and others were nocturnal. Furthermore, some of them displayed an extraordinary flexibility in phasing because they were light active but dark feeding, and vice versa. Generally, goldfish tended to be day active, although their feeding habits appeared equally distributed between light and dark phases. Under DD, goldfish showed free-running rhythms that averaged 25.3 +/- 1.8 h and 24.4 +/- 1.7 h for locomotor activity and feeding, respectively, but that were slightly shorter under LD pulses. These results indicate that the type of phasing of locomotor activity did not necessarily decide the feeding phase; much of this is explained by the fact that goldfish were self-fed. Flexibility in phasing and a certain degree of independence between locomotor and feeding activities could be seen as an adaptative response of the highly adaptable circadian system of fish.

Animals↗

Nerve growth factor (NGF) induces increase in multi-ubiquitin chains and concomitant decrease in free ubiquitin in nuclei of PC12h.

Changes in Ubiquitin-immunoreactivity after nerve growth factor (NGF) treatment were investigated in PC12h cells. Ubiquitin-immunoreactivity was increased in the nucleus of NGF-treated cells. The quantitative analysis revealed that, after 7 days of NGF treatment, almost 20% of cells had ubiquitin-immunoreactive nuclei and the frequency was increased thereafter. Levels of free ubiquitin and multi-ubiquitin chains were measured by radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA), respectively. Measurements were carried out for four subcellular fractions: urea- and water-soluble extracts of nuclei and cytoplasm. Decrease in free ubiquitin was observed in water-soluble cytoplasmic extracts of NGF-treated cells, though increase in multi-ubiquitin chains in the same fraction was not observed. As for nuclei, increase in multi-ubiquitin chains and concomitant decrease in free ubiquitin were found in the water-soluble extracts after NGF treatment. Levels of multi-ubiquitin chains did not change in urea-soluble cytoplasmic extracts as well as nuclear urea-soluble ones after NGF treatment. These results indicated that multi-ubiquitination of nuclear proteins is increased during NGF-induced neuronal differentiation of PC12h cells.

Animals↗

Decreased levels of 2-amino-3-methylimidazo[4,5-f]quinoline-DNA adducts in rats treated with beta-carotene, alpha-tocopherol and freeze-dried aloe.

To assess mechanisms of chemoprevention of hepatocarcinogenesis by trans-beta-carotene (beta-C), DL-alpha-tocopherol (alpha-T), and freeze-dried whole leaves of Kidachi aloe (Aloe), formation of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ)-DNA adducts was measured by 32P-post-labeling analysis, and CYP1A1 and CYP1A2 protein levels were analyzed by ELISA. Group 1 rats were fed diet containing 0.02% beta-C, 1.5% alpha-T or 30% Aloe over an 8-day period, while group 2 was given basal diet alone. On day 7, all animals were subjected to two-thirds partial hepatectomy (PH). Twelve hours after PH, they received a single dose of the carcinogenic food pyrolysate IQ (100 mg/kg) intragastrically, to initiate hepatocarcinogenesis. Rats were killed 6, 12, 24 and 48 h after IQ administration. The levels of adducts, expressed as relative adduct labeling values in rats treated with beta-C, alpha-T and Aloe, were decreased as compared with the control group at hour 24 (36 h after PH), with a significant difference in the case of the beta-C group (46.4% of the control value). Similarly, all showed a tendency for decrease at hour 48. Furthermore, the levels of CYP1A2, known to be responsible for activation of IQ, showed a significant reduction at hour 24. It is concluded that beta-C, and possibly also alpha-T and Aloe, have the potential to reduce IQ-DNA adduct formation, presumably as a result of decreased formation of active metabolites. The results may explain, at least in part, the previously observed inhibitory effects of these compounds on induction of preneoplastic hepatocellular lesions.

Aloe↗

Demonstration of organotropic effects of chemopreventive agents in multiorgan carcinogenesis models.

Organotropic chemopreventive effects of three (pro)vitamins and three unsaturated fatty acids were examined using mouse and rat multiorgan carcinogenesis models. For the study of (pro)vitamins, male and female B6C3F1 mice were treated with N,N-diethylnitrosamine (DEN) and N-methyl-N-nitrosourea (MNU) during the first 11 weeks, then from weeks 12 to 32 they received alpha-carotene (0.4 mg/mouse), beta-carotene (0.4 mg/mouse) or alpha-tocopherol (40 mg/mouse) three times a week by gavage; control mice received vehicle alone. In male mice, alpha-carotene significantly reduced liver weights, representing a reduced tumour mass (P < 0.001), and alpha-carotene, beta-carotene and alpha-tocopherol significantly reduced the numbers of liver tumours (adenomas and carcinomas combined) (P < 0.001-0.01) as compared with control mice, the effects being greatest with alpha-carotene. In female mice, alpha-carotene significantly decreased the number of liver tumours (P < 0.001). In the lung, alpha-carotene and alpha-tocopherol reduced the area of lesions (hyperplasias and adenomas combined) only in males (P < 0.05). For the study of unsaturated fatty acids, F344 male rats were treated with DEN, MNU, N-butyl-N-hydroxybutylnitrosamine (BBN), 1,2-dimethylhydrazine (DMH) and N,N-bis(2-hydroxy)propylnitrosamine during the first 5 weeks, then from weeks 6 to 36 they were given docosahexaenoic acid (C22:6), eicosapentaenoic acid (C20:5) or linoleic acid (C18:2) at 1.0 g/rat, three times a week by gavage; control rats were treated with oleic acid (C18:1) using the same protocol. All animals were fed a low linoleic acid and calorie-adjusted basal diet during fatty acid administration. Docosahexaenoic acid and linoleic acid reduced tumours in the large and small intestines, respectively. However, they did not influence the yield of preneoplastic liver, lung, kidney, forestomach and urinary bladder lesions. The data thus provide evidence for organotropic effects of carotenoids and unsaturated fatty acids on carcinogenesis.

Animals↗

Effects of pinealectomy and constant light exposure on day-night changes of melatonin binding sites in the goldfish brain.

Effects of pinealectomy and constant light exposure on day-night changes of melatonin binding sites in the goldfish brain were examined. The density and affinity of binding sites were higher at mid-day than at mid-night in sham-pinealectomized goldfish under light-dark cycles. The rhythms disappeared after pinealectomy, or constant light exposure both of which abolish plasma melatonin rhythms. The effects of pinealectomy and constant light exposure were not additive. These results indicate that diel changes of melatonin binding sites in the goldfish brain are regulated by endogenous melatonin of pineal origin.

Animals↗

Characterization of extrathymic T cells induced by recombinant murine interferon-beta in the peritoneal cavity of nude mice.

The effects of recombinant murine interferon-beta (rMuIFN-beta) on extrathymic lymphocyte formation in the peritoneal cavity of BALB/c athymic nude mice and tumor-bearing nude mice were examined for comparison with BALB/c normal euthymic mice. The 7 days of administration of 1 x 10(5) IU rMuIFN-beta caused the number of these cells to increase remarkably in the peritoneal cavity as a unique subset of asialoGM1+CD4+CD8-TcR alpha beta+ T cells. The asialoGM1+CD4+ T cells produced IL-2 and IFN-gamma in primary culture without stimulant but did not proliferate. Thus, extrathymic T cells can be induced easily in the peritoneal cavity in addition to the thymus for host defense systems.

Adenocarcinoma↗

Characteristics of the inhibitory effect of mitoxantrone and pirarubicin on lung metastases of colon carcinoma 26.

This study was performed to evaluate the antimetastatic activity of antitumor agents against metastatic colon carcinoma 26 (Co 26Lu), and to investigate their mechanisms of action. Pirarubicin demonstrated the most striking antitumor activity in mice bearing intravenously injected Co 26Lu cells. Etoposide and mitoxantrone also showed marked antitumor activity. Pirarubicin and mitoxantrone also exerted remarkable inhibitory effect on spontaneous lung metastases from subcutaneously implanted Co 26Lu. Pirarubicin showed marked inhibition of both primary tumor growth and lung metastases. Mitoxantrone was effective in preventing lung metastases even at doses that did not exhibit an antitumor effect on the primary tumor. Moreover, mitoxantrone administered two days after intravenous injection of tumor cells obviously reduced the number of lung colonies, while simultaneous injection of the drug did not inhibit colony formation. Mitoxantrone effectively inhibited angiogenesis on the chorioallantoic membrane at doses that did not affect the growth rate of embryos. These results suggest that mitoxantrone, besides its direct antitumor effect on tumor cells, may inhibit lung metastases by inhibiting angiogenesis.

Animals↗