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Biomedical subjects

E Suzuki

Publications and source records attributed to E Suzuki.

At least 91 records · Page 5Linked to original sources
PubMed2000

Investigation on the formation and the determination of gamma-glutamyl-beta-alanylhistidine and related isopeptide in the macromolecular fraction of beef soup stock.

To confirm the formation of gamma-glutamyl-beta-alanylhistidine and related peptide, a model solution (amide-containing amino acids and carnosine) has been heated, and the products are investigated. Spectroscopical analysis indicates that the major product from asparagine and carnosine is beta-aspartyl-beta-alanylhistidine, and that from glutamine and carnosine is gamma-glutamyl-beta-alanylhistidine. Furthermore, to confirm the increase of the above peptides during the heating process of food, an HPLC method for the determination of these isopeptides in food protein is constructed. The isopeptides are liberated by proteolytic digestion and fractionated by solid-phase extraction using Toyopack IC-SP cartridges. The fraction containing the isopeptides is derivatized with phenylisothiocyanate (PITC) and separated and quantified by HPLC using an octadecyl-silica column. As a result of quantification, an increase of the gamma-glutamyl-beta-alanylhistidine isopeptide in the macromolecular fraction of heated beef soup stock solution has been observed. These results suggest that the formation of the isopeptide occurs in the heating of various foods containing carnosine.

Chromatography, High Pressure Liquid↗
PubMed2000

Relationship between the glass transition of soy protein and molecular structure.

The change in molecular structure of the soy protein samples as a result of the microbial transglutaminase treatment was studied using solid-state (13)C NMR spectroscopy and circular dichroism (CD), and the relation to the glass transition temperature (T(g)) was examined. From NMR measurements, the structure of the local region of the C(alpha) methine was observed to change, and the region had relatively high mobility. From CD measurements, the structural change seemed to be caused by the change in the secondary structure (disintegration of the beta-structure). By comparison with the T(g) of another protein, the state of the secondary structure of a protein was suggested to be a key in determining its T(g).

Circular Dichroism↗
PubMed2000

Quantitation of catechol estrogens and their N-acetylcysteine conjugates in urine of rats and hamsters.

A method for the analysis of N-acetylcysteine conjugates of catechol estrogens [catechol estrogen mercapturates (CE SRs)], which are likely to be urinary markers of estrogen-induced tumors, was established in this study. The characteristics of the method that was established were (1) cleanup of urine using the immunoaffinity column of CE SRs, (2) detection of catechol estrogens (CEs) and CE SRs by electrochemical detection, which provided the high specificity, and (3) stability of CE SRs through the cleanup. Using this method, the simultaneous quantitation of 2-hydroxy-17beta-estradiol (2-OHE(2)), 4-hydroxy-17beta-estradiol (4-OHE(2)), 2-hydroxyestrone (2-OHE(1)), 4-hydroxyestrone (4-OHE(1)), 2-hydroxyestrone 1-N-acetylcysteine thioether (2-OHE(1) 1SR), 2-hydroxyestrone 4-N-acetylcysteine thioether (2-OHE(1) 4SR), and 4-hydroxyestrone 2-N-acetylcysteine thioether (4-OHE(1) 2SR) in the range of 1-15 ng was performed. We first demonstrated the presence of CE SRs, 2-OHE(1) 1SR and 2-OHE(1) 4SR, in urine from rats treated intraperitoneally with 17beta-estradiol (E(2)) at a dose of 5 mg/kg. In female rats, the amount of 2-OHE(1) 1SR was several-fold greater than that of 2-OHE(1) 4SR, while the presence of 4-OHE(1) 2SR was not confirmed. The level of CEs and CE SRs in male rats was approximately (1)/(2)-(1)/(20) of that in female rats. The excretion rate following administration of 2-OHE(1) at 2 mg/kg and that following the administration of 4-OHE(1) at 2 mg/kg were different in female rats. In addition, 4-OHE(1) 2SR was present in the urine of male Syrian hamsters treated intraperitoneally with E(2), whereas it was absent in rats.

Acetylcysteine↗
PubMed2000

Postsynaptic filopodia in muscle cells interact with innervating motoneuron axons.

Precise synaptogenesis is crucial to brain development, and depends on the ability of specific partner cells to locate and communicate with one another. Dynamic properties of axonal filopodia during synaptic targeting are well documented, but the cytomorphological dynamics of postsynaptic cells have received less attention. In Drosophila embryos, muscle cells bear numerous postsynaptic filopodia ('myopodia') during motoneuron targeting. Here we show that myopodia are actin-filled microprocesses, which progressively clustered at the site of motoneuron innervation while intermingling with presynaptic filopodia. In prospero mutants, which have severe delays in axon outgrowth from the CNS, myopodia were present initially but clustering behavior was not observed, demonstrating that clustering depends on innervating axons. Thus, postsynaptic filopodia are capable of intimate interaction with innervating presynaptic axons. We propose that, by contributing to direct long-distance cellular communication, they are dynamically involved in synaptic matchmaking.

Actins↗
PubMed2000

A phase I dose escalation study of multicyclic, dose-intensive chemotherapy with peripheral blood stem cell support for small cell lung cancer.

A phase I dose-escalation study of multicyclic, ifosfamide, carboplatin, and etoposide (ICE) with sequential reinfusion of peripheral blood stem cells (PBSCs) was conducted to determine the maximum-tolerated dose (MTD) of ICE. Twenty-four patients with SCLC (LD: 6, ED: 18) were treated with ifosfamide (3000-9000 mg/m2, 24-h infusion), carboplatin (300-400 mg/m2), and etoposide (300 mg/m2) followed by subcutaneous filgrastim (75 microg/day) from day 4 to the day of PBSC collection. PBSC were harvested when the WBC count reached >/=5 x 109/l. The leukapheresis product was cryopreserved and reinfused on day 4 of the next cycle, which was started 48 h after the last PBSC collection. The ifosfamide dose was escalated as follows: 3000 mg/m2 (level 1), 5000 mg/m2 (level 2), 7000 mg/m2 (level 3), 9000 mg/m2 (level 4). Patients with LD were treated with concurrent radiotherapy at 1.5 Gy twice daily for the initial 3 weeks to a total dose of 45 Gy and MTD, defined separately. Patients were evaluated for hematologic and non-hematologic toxicity, actual dose intensities, as well as response to therapy. The maximum-tolerated dose (MTD) was defined as the dose level at which more than 5 days of grade 4 myelo- suppression or non-hematologic toxicity greater than grade 3 developed in two thirds of the patients. For ED cases, MTD was level 4 and the recommended dose of ifosfamide was 7000 mg/m2. For LD cases, the recommended dose of ifosfamide was 5000 mg/m2. The dose limiting toxicity of multicyclic ICE was hemato- logic toxicity and CNS toxicity which manifested as ataxia. Tumor responses were seen in all patients, with 14 patients showing a complete response. The actual total dose-intensity at the recommended dose level was 2.2 and 1.74, for ED and LD, respectively, compared with previously reported ICE regimens. PBSC support for dose-intensive ICE regimen permitted dose escalation of ifosfamide with a mean interval of 16-17 days. We conclude that this regimen is well tolerated, with acceptable hematological and non-hematological toxicity. Bone Marrow Transplantation (2000) 25, 5-11.

Adult↗
PubMed2000

Adrenomedullin and nitric oxide inhibit human endothelial cell apoptosis via a cyclic GMP-independent mechanism.

Adrenomedullin, which was discovered as a vasodilating peptide, has been reported to be produced in various organs, in which adrenomedullin regulates not only vascular tone but also cell proliferation and differentiation in an autocrine/paracrine manner. We evaluated the effect of adrenomedullin on endothelial cell apoptosis. Human umbilical vein endothelial cells underwent apoptosis when cultured in serum-free medium. Treatment with adrenomedullin reduced the number of cells with pyknotic nuclei (Hoechst 33258 staining) and inhibited cell death (dimethylthiazol-diphenyltetrazolium bromide assay) in a dose-dependent manner. The administration of adrenomedullin did not alter the expression levels of Bcl-2 family proteins. Experiments with analogs of cAMP or a cAMP-elevating agonist demonstrated that elevation of the intracellular cAMP concentration does not mediate the antiapoptotic effect of adrenomedullin. The coadministration of N-nitro-L-arginine methyl ester (2 mmol/L), an inhibitor of nitric oxide synthase, abrogated the effect of adrenomedullin. Lower doses of sodium nitroprusside (1 to 10 micromol/L), a nitric oxide donor, mimicked the antiapoptotic effect of adrenomedullin. The antiapoptotic effect of sodium nitroprusside was not attenuated by the inhibition of soluble guanylyl cyclase with 1 micromol/L oxadiazolo-quinoxalin-1-one nor could apoptosis be inhibited by the incubation of human umbilical vein endothelial cells with 1 mmol/L 8-bromo-cGMP, a cell-permeant cGMP analog. These results indicate that adrenomedullin and nitric oxide inhibit endothelial cell apoptosis via a cGMP-independent mechanism.

Adrenomedullin↗
PubMed2000

Synaptic development is controlled in the periactive zones of Drosophila synapses.

A cell-adhesion molecule fasciclin 2 (FAS2), which is required for synaptic growth and still life (SIF), an activator of RAC, were found to localize in the surrounding region of the active zone, defining the periactive zone in Drosophila neuromuscular synapses. BetaPS integrin and discs large (DLG), both involved in synaptic development, also decorated the zone. However, shibire (SHI), the Drosophila dynamin that regulates endocytosis, was found in the distinct region. Mutant analyses showed that sif genetically interacted with Fas2 in synaptic growth and that the proper localization of SIF required FAS2, suggesting that they are components in related signaling pathways that locally function in the periactive zones. We propose that neurotransmission and synaptic growth are primarily regulated in segregated subcellular spaces, active zones and periactive zones, respectively.

Animals↗
PubMed2000

Temporally restricted expression of transcription factor betaFTZ-F1: significance for embryogenesis, molting and metamorphosis in Drosophila melanogaster.

FTZ-F1, a member of the nuclear receptor superfamily, has been implicated in the activation of the segmentation gene fushi tarazu during early embryogenesis of Drosophila melanogaster. We found that an isoform of FTZ-F1, betaFTZ-F1, is expressed in the nuclei of almost all tissues slightly before the first and second larval ecdysis and before pupation. Severely affected ftz-f1 mutants display an embryonic lethal phenotype, but can be rescued by ectopic expression of betaFTZ-F1 during the period of endogenous betaFTZ-F1 expression in the wild type. The resulting larvae are not able to molt, but this activity is rescued again by forced expression of betaFTZ-F1, allowing progression to the next larval instar stage. On the other hand, premature expression of betaFTZ-F1 in wild-type larvae at mid-first instar or mid-second instar stages causes defects in the molting process. Sensitive periods were found to be around the time of peak ecdysteroid levels and slightly before the start of endogenous betaFTZ-F1 expression. A hypomorphic ftz-f1 mutant that arrests in the prepupal stage can also be rescued by ectopic, time-specific expression of betaFTZ-F1. Failure of salivary gland histolysis, one of the phenotypes of the ftz-f1 mutant, is rescued by forced expression of the ftz-f1 downstream gene BR-C during the late prepupal period. These results suggest that betaFTZ-F1 regulates genes associated with ecdysis and metamorphosis, and that the exact timing of its action in the ecdysone-induced gene cascade is important for proper development.

Animals↗
PubMed2000

Intermediate state during the crystal transition in aspartame, studied with thermal analysis, solid-state NMR, and molecular dynamics simulation.

Aspartame (L-alpha-aspartyl-L-phenylalanine methyl ester) is a dipeptide sweetener about 200 times as sweet as sugar. It exists in crystal forms such as IA, IB, IIA, and IIB, which differ in crystal structure and in the degree of hydration. Among these, IIA is the most stable crystal form, and its crystal structure has been well determined (Hatada et al., J. Am. Chem. Soc., 107, 4279-4282 (1985)). To elucidate the structural factors of thermal stability in the IIA form of aspartame and to examine the physical process in the crystal transformation between the IIA and IIB forms, we performed a thermal analysis and solid-state NMR measurements. We found that a quasi-stable intermediate state exists in the transformation, and it has the same crystal lattice as the usual IIA form, despite the dehydration from 1/2 mol to 1/3 mol per 1 mol of aspartame. The results of the energy component analysis and the molecular dynamics simulation suggest that the entropic effect promotes the generation of the intermediate state, which is presumably caused by the evaporation of the water of crystallization and the increase of molecular motion in aspartame. Thus, the thermal stability of the IIA form is attributable to a structural property, i.e., the crystal lattice itself is retained during the above dehydration. Moreover, the molecular dynamics simulations suggest that the aspartame molecules have two kinds of conformational flexibility in the intermediate state.

Aspartame↗
PubMed2000

Effects of vasodilatory antihypertensive agents on endothelial dysfunction in rats with ischemic acute renal failure.

Ischemic acute renal failure is associated with vascular endothelial dysfunction. We examined whether vasodilatory antihypertensive agents would improve endothelial function in rats with ischemia/reperfusion renal injury. Rat kidneys were isolated and perfused after clipping of the bilateral renal arteries for 45 min and reperfusion for 24 h, and renal perfusion pressure and nitric oxide concentration in the venous effluent (chemiluminescence assay) were monitored. Preischemic administration of celiprolol (a beta-blocker; 100 mg/kg p.o.), benidipine (a calcium channel blocker; 1 mg/kg p.o.), or imidapril (an angiotensin converting-enzyme inhibitor; 3 mg/kg p.o.) restored endothelial function in rats subjected to acute renal ischemia (deltarenal perfusion pressure [10(-8) M acetylcholine]: sham -42+/-3%, ischemia -31+/-1%, ischemia +celiprolol -39+/-1%*, ischemia+benidipine -38+/-2%*, ischemia+imidapril -42+/-2%*; *p<0.05 vs. ischemia). Serum urea nitrogen and creatinine levels were also lower in the treated groups. Furthermore, ischemia-induced decreases in the response to acetylcholine and renal excretory function were smaller in SHR than in deoxycorticosterone-salt hypertensive rats, in which endothelial damage was marked. These results suggest that preischemic endothelial function may influence the degree of ischemic renal injury. Calcium channel blockers, converting-enzyme inhibitors, and endothelial NO synthase-activating beta-blockers had beneficial effects on renovascular endothelial dysfunction due to ischemia.

Acetylcholine↗
PubMed2000

The presence of specialized epithelial cells on the bronchus-associated lymphoid tissue (BALT) in the mouse.

The aggregation of lymphoid cells in the bronchial mucosa has been named the bronchus-associated lymphoid tissue (BALT) and investigated in comparison with the gut-associated lymphoid tissue (GALT). To elucidate precisely the structure and function of the BALT, the present study examined the age-related change in the mouse BALT by light microscopy. We also observed the characteristics of the overlying epithelium, especially the lectin-binding properties of the epithelial cells, by the combined use of light microscopy (LM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). By LM, lymphoid aggregates were not recognizable in the bronchial mucosa of young (8-10 week-old) mice, while they were commonly found at the second to fourth branching portions of the bronchial tree in older (32-40 week-old) mice. The epithelium overlying the lymphoid aggregates of the mature mice often contained a large number of mononuclear cells. Lectin cytochemistry revealed that UEA1 (Ulex europaeus agglutinin 1) positive cells were not only restricted to the overlying epithelium of the BALT in the older mice but also found in a cell group in the mucous epithelium at the branching portions in the young mice. Comparison between the LM and SEM images of the UEA1-stained whole mount specimens clarified the surface morphology of the lectin-stained epithelial cells, showing them to be non-ciliated cells with a large number of short microvillous projections on the apical surface. TEM studies further demonstrated that the UEA1 reaction products appeared on the plasma membrane of the non-ciliated cells which often enfolded lymphocytes in the old mice. Latex microbeads, which were administrated intratracheally, were selectively taken up by the UEA1-positive cells of the BALT. These results indicate that the mouse BALT has specialized epithelial cells similar to the UEA1 positive M cells in the GALT and probably functions as a part of the mucosal immune system. This study also showed the possibility that the UEA1 positive cells appear in the mucous epithelium before the formation of the BALT.

Age Factors↗
PubMed2000

Time-dependent changes of tissue erythorbic acid concentrations in guinea pigs.

The concentrations of erythorbic acid(ErA) and L-ascorbic acid(AsA) in the tissues of guinea pigs orally administered AsA or ErA were measured over the passage of time using high-performance liquid chromatography. Guinea pigs were each administered 5 mg AsA or 100 mg ErA, and killed at a specified time thereafter. The AsA concentrations in the tissues of AsA-administered guinea pigs and the ErA concentrations in the tissues of ErA-administered guinea pigs increased for 3 h after the respective administrations and decreased thereafter in both groups. The AsA concentration in the tissues of AsA-administered guinea pigs tended to be similar to the sum of AsA and ErA concentrations in the ErA-administered guinea pigs within 3 h after administration.

Adrenal Glands↗
PubMed2000

Enhanced expression of mRNA coding for the adrenaline-synthesizing enzyme phenylethanolamine-N-methyl transferase in adrenaline-secreting pheochromocytomas.

PURPOSE: In some pheochromocytomas, the tumors contain and secrete greater amounts of adrenaline than do normal adrenal medullas. It is not yet known how adrenaline synthesis is enhanced in the adrenaline-secreting pheochromocytomas. MATERIALS AND METHODS: As a first step toward understanding the molecular mechanisms by which adrenaline synthesis is controlled in these tumors, we measured the level of mRNA coding for the adrenaline-synthesizing enzyme phenylethanolamine N-methyl transferase (PNMT) and the content of adrenaline in the pheochromocytomas (n = 9), including 3 cases of the adrenaline-secreting type (one of the patients had bilateral pheochromocytomas), and in normal adrenal medullas (n = 7). We then measured the concentration of cortisol, which is thought to regulate the PNMT activity. Finally, we examined the expression of the mRNA for Egr-1, which was recently reported to be a transcriptional factor regulating PNMT gene expression. RESULTS: In the 4 tissue specimens from 3 adrenaline-secreting pheochromocytomas, the contents of adrenaline and the PNMT mRNA expression were considerably greater than those of the normal adrenal medullas. PNMT immunoreactivity was only detected in the adrenaline-secreting tumors. Three of the 4 specimens showed high concentrations of cortisol. To show the capacity for cortisol production locally in the pheochromocytoma tissues, we showed the expression of a glucocorticoid biosynthetic enzyme, 17alpha-hydroxylase, in the tumors by Western blotting. PNMT expression was found to be associated with 17alpha-hydroxylase expression in the tumors. The glucocorticoid receptor expression was also correlated with PNMT expression in the tumors and the expression of Egr-1 was also high in 3 of the 4 specimens. CONCLUSIONS: These findings indicate that adrenaline production in adrenaline-secreting pheochromocytomas is primarily controlled by the level of PNMT gene expression, and that the gene expression may be enhanced by both cortisol and Egr-1.

Adrenal Gland Neoplasms↗
PubMed2000

[Multiple nodular and patchy shadows in chest radiograph in a patient with autoimmune hemolytic anemia associated with chronic thyroiditis].

A 65-year-old woman was admitted because of dyspnea and persistent cough. Her chest radiograph and CT films revealed multiple nodular and patchy shadows in bilateral lungs, indicating bronchiolitis obliterans organizing pneumonia (BOOP)-like shadows. Autoimmune hemolytic anemia was diagnosed because peripheral blood cell analysis revealed anemia (hemoglobin, 5.5 g/dl) with increased reticulocytes (253/1000), and positive direct and indirect Coomb's tests. In addition, decreased diffusion capacity and mild hypoxemia (PaO2, 71 Torr) were recognized. Bronchoalveolar lavage fluid (BALF) showed the increased number of total cells and lymphocytes, and a decreased CD4(helper)/CD8(suppressor) ratio (= 0.65). Histopathological features of transbronchial lung biopsy showed an infiltration of lymphocytes in the alveolar walls and spaces, and thickening of the alveolar walls. A discrepancy was apparent among roentgenogram, histopathological findings and BALF results. Open lung biopsy had not been performed because of severe hemolytic anemia and lack of informed consent. In addition, the patient suffered from chronic thyroiditis with positive thyroid autoantibody. After oral administration of prednisolone, both pulmonary shadows and anemia improved. We concluded that autoimmune hemolytic anemia may be associated with steroid-sensitive interstitial lung lesions in some cases. Further immunologic studies are necessary to elucidate the relationship between autoimmune hemolytic anemia and interstitial lung shadows.

Aged↗
PubMed2000

[Myeloperoxidase-antineutrophil cytoplasmic antibody positive alveolar hemorrhage during propylthiouracil therapy for hyperthyroidism].

A 62-year-old woman had been treated with propylthiouracil(PTU) for hyperthyroidism. Because bloody sputum, dyspnea, and severe hypoxemia developed, the patient was admitted to our hospital. Chest X-ray and chest computed tomographic (CT) films disclosed diffuse infiltrative shadows in both lung fields. Bronchoalveolar lavage revealed abundant hemosiderin-laden macrophages. Alveolar hemorrhage associated with myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA) positive vasculitis syndrome was diagnosed because of the high serum level of MPO-ANCA. After the initiation of steroid therapy and termination of PTU, the infiltrative shadows in both lung fields disappeared, PaO2 improved, and MPO-ANCA decreased. There have been some reports of MPO-ANCA positive vasculitis syndrome developing during PTU therapy, but most were concerned with renal disease. We concluded that PTU and similar agents should be given consideration as one of the possible causes of MPO-ANCA-induced alveolar hemorrhage.

Antibodies, Antineutrophil Cytoplasmic↗
PubMed2000

[Levofloxacin-induced eosinophilic pneumonia complicated by bronchial asthma].

A 76-year-old woman who had complained of cough and productive sputum since mid-January, 1999, was admitted to our hospital with fever and dyspnea on February 4, 1999. She had been treated with levofloxacin at an outpatient clinic. On admission, she had orthopnea, and auscultation revealed coarse crackles and wheeze in the bilateral lung fields. Chest x-ray and CT films showed non-segmental infiltration in bilateral lung fields. Laboratory data revealed eosinophilia in peripheral blood (= 24%) and sputum (= 10%), airflow limitation, hypoxemia (PaO2: 46 Torr), and increased airway responsiveness to methacholine (Dmin: 0.127 units). A bronchoalveolar lavage (BAL) fluid showed increased total cells and a 55% increase in eosinophils, and CD4/CD8 ratio was decreased to 0.8. In addition, IL-5 was increased in BAL fluid. Transbronchial lung biopsy specimens revealed infiltrations of eosinophils in the alveolar and interstitial compartments. Histological features of the bronchial biopsy specimens included increased eosinophils in the submucosa and goblet cell metaplasia. The woman was diagnosed with eosinophilic pneumonia complicated by bronchial asthma. She was given theophylline, pranlukast hydrate, and an inhaled beta 2 receptor agonist (procaterol hydrochloride), and pre-admission drugs including Levofloxacin were discontinued. Her symptoms were improved, peak expiratory flow rate and PaO2 increased, airway responsiveness to methacholine decreased (Dmin: 0.615 units), and radiographic abnormalities disappeared without steroid therapy. A leukocyte migration test for levofloxacin was weakly positive. An environmental provocation test in the patient's home gave negative results. A challenge test for levofloxacin was not performed due to a lack of informed consent. Based on these findings, we diagnosed this case as levofloxacin-induced lung injury manifesting as eosinophilic pneumonia complicated by bronchial asthma. Levofloxacin should be added to the list of agents that can produce eosinophilic pneumonia.

Aged↗
PubMed2000

[Efficacy of sustained-released theophylline on peak expiratory flow rate in asthmatic patients].

Nocturnal worsening is an important problem in asthma management. We evaluated the efficacy of two sustained-released theophylline formulations, administered twice-a-day (TD) and once-a-day (UP). In 20 asthmatic patients with low peak expiratory flow rate (PEF) in the morning (< 80% of predicted) administered TD (mean dose 475 mg/day) for 2 weeks, we evaluated PEF, diurnal variability of PEF, symptom score, results of spirometry and serum theophylline concentration (STC) and then changed theophylline, from TD to UP (mean dose 470 mg/day) for 2-4 weeks. PEF in the morning was higher during the UP period (mean +/- SD: 335 +/- 110 L/min) than during the TD period (mean +/- SD: 308 +/- 95 L/min) (p < 0.05), and diurnal variabirity of PEF was lower during the UP period (17.1 +/- 8.0%) than TD period (22.9 +/- 13.4%) (p < 0.05). However, there were no changes in PEF at night or daytime, symptom score, spirometry parameters or STC during the study period. The improvement in morning PEF observed in this study was consistent with the differences in pharmacokinetics between the two theophylline formulations. Morning PEF was increased during the UP period, probably because STC in the early morning during the UP period was higher than STC during the TD period.

Adult↗