Biomedical subjects

Y Doi

Publications and source records attributed to Y Doi.

At least 91 records · Page 5Linked to original sources

PubMed2000

Job strain, Type A behavior pattern, and the prevalence of coronary atherosclerosis in Japanese working men.

OBJECTIVE: To examine the relation of type A behavior pattern and job strain to angiographically documented coronary stenosis. METHODS: Subjects were 197 male Japanese patients with a full-time job. A questionnaire-based interview elicited psychosocial and other factors. Type A behavior pattern was measured by 12 questions, and job strain by the method of Karasek. Significant coronary stenosis was defined when a 75% or greater luminal narrowing occurred at one or more major coronary arteries or when a 50% or greater narrowing occurred at the left main artery. Logistic regression analysis was used to calculate odds ratio (OR) and 95% confidence interval (CI) with adjustment for traditional coronary risk factors and job type. RESULTS: Type A behavior pattern was related to a statistically non-significant lower prevalence of the coronary stenosis especially in the absence of job strain (adjusted OR 0.6, 95% CI 0.3-1.2). Job strain was non-significantly associated with a modestly increased prevalence of coronary stenosis (OR 1.7, 95% CI 0.6-5.2). CONCLUSION: These findings suggest that both the behavioral pattern and psychosocial work environment may be related to coronary artery stenosis.

Adult↗

PubMed2000

Troglitazone enhances glycolysis and improves intracellular glucose metabolism in rat mesangial cells.

To clarify the action of the new antidiabetic agent, troglitazone, on rat mesangial cells, we assessed its effect on the uptake and intracellular metabolism of glucose. Troglitazone increased the uptake of 2-deoxyglucose (2DOG) in a dose-dependent manner with an upregulation of glucose transporter 1 (GLUT1) mRNA, whereas it had no effect on the uptake of alpha-methyl glucoside (AMG). This troglitazone-induced glucose uptake was not suppressed by phlorizin. In 5 mmol/L glucose, 2 microg/mL (4.5 micromol/L) troglitazone increased glucose consumption 2.9-fold, similar to that in 20 mmol/L glucose. Troglitazone increased the production of pyruvate and lactate as a consequence of the increase in glycolysis, but did not increase the cellular ATP content. Troglitazone improved the high-glucose-induced accumulation of intracellular sorbitol and fructose and elevation of the cellular redox potential. These data suggest that troglitazone enhances glucose uptake through GLUT1 with an acceleration of glycolysis, and improves the abnormal intracellular glucose metabolism under high-glucose conditions in rat mesangial cells.

Adenosine Triphosphate↗

PubMed2000

Miltpain, a cysteine proteinase, from milt of Pacific cod (Gadus macrocephalus): purification and characterization.

Miltpain (EC.3.4.22.-) is a cysteine proteinase that preferentially hydrolyzes basic proteins, previously found in the milt of chum salmon. Here we report a similar cysteine proteinase in the milt of the marine Pacific cod. The enzyme was isolated and purified 6900-fold and with an estimated mass of 63 kDa by gel filtration chromatography and 72 kDa by SDS/PAGE. Cod miltpain has an optimum pH of 6.0 for Z-Arg-Arg-MCA hydrolysis, and Km of 11.5 microM and kcat of 19.0 s-1 with Z-Arg-Arg-MCA. It requires a thiol-inducing reagent for activation and is inhibited by E-64, iodoacetamide, CA-074, PCMB, NEM, TLCK, TPCK, ZPCK and o-phenanthroline. This proteinase strongly hydrolyzes basic proteins such as salmine, clupeine and histone, and exhibits unique substrate specificity toward paired basic residues such as Lys-Arg, Arg-Arg on the substrates of P2-P1. The isoelectric point is 5.2 by isoelectric focusing. N-Terminal sequencing gave a sequence of < EVPVEVVRXYVTSAPEK. The cysteine proteinase from Pacific cod very closely matches the previously reported miltpain from chum salmon.

Amino Acid Sequence↗

PubMed2000

Relation between green tea consumption and the severity of coronary atherosclerosis among Japanese men and women.

PURPOSE: To examine the relation between green tea consumption and arteriographically determined coronary atherosclerosis. METHODS: Study subjects were 512 patients (302 men and 210 women) aged 30 years or older who underwent coronary arteriography for the first time at four hospitals in Fukuoka City or one hospital in an adjacent city between September 1996 and August 1997. Lifestyle characteristics including green tea consumption were ascertained before arteriography by a questionnaire supported with interview. RESULTS: 117 men (38.7%) and 50 women (23.8%) had significant stenosis of one or more coronary arteries. Green tea consumption tended to be inversely associated with coronary atherosclerosis in men, but not in women. An evident, protective association between green tea and coronary atherosclerosis was observed in a subgroup of 262 men excluding those under dietary or drug treatment for diabetes mellitus. In this subgroup, after adjustment for traditional coronary risk factors and coffee, odds ratios of significant stenosis for consumption of 2-3 cups and 4 or more cups per day were 0.5 (95% confidence interval 0.2-1.2) and 0.4 (0.2-0.9), respectively, as compared with a consumption of one cup per day or less. CONCLUSIONS: The results indicate that green tea may be protective against coronary atherosclerosis at least in men.

Adult↗

PubMed2000

Transient expression of activin betaA mRNA on osteoprogenitor cells in rat bone regeneration after drill-hole injury.

We investigated the expression of activin betaA on osteoprogenitor cells in the regenerating bone and bone marrow of the rat femur after drill-hole injury, by immunocytochemistry and in situ hybridization. The periosteum and endosteum adjacent to the wound region showed marked thickening at day 3 and abundant osteoprogenitor cells, which were immunoreactive for proliferating cell nuclear antigen and showed positive reactions for alkaline phosphatase activity, and existed in the inner layer of the periosteum as well as in the endosteum. During the same period, these osteoprogenitor cells began to exhibit activin betaA immunoreactivity and mRNA expression. However, the latter expression gradually reduced the intensity as the cells started to express osteocalcin mRNA during their differentiation to osteoblasts participating in the periosteal and medullary bone formation from day 5. Immunoreactivity for activin type IB and II receptors was also found on activin betaA-immunoreactive cells between days 3 and 7. The above findings suggest that proliferating osteoprogenitor cells, before their transformation to osteoblasts, transiently produce and release activin A, which may play crucial roles in bone and bone marrow regeneration in a receptor-mediated, autocrine and paracrine fashion.

Animals↗

PubMed2000

Function of the catalytic domain of poly(3-hydroxybutyrate) depolymerase from Pseudomonas stutzeri.

The mechanism of enzymatic hydrolysis for (R)-3-hydroxybutyrate (3HB) oligomers with poly[(R)-3-hydroxybutyrate] [P(3HB)] depolymerase (PhaZpst) from Pseudomonas stutzeri was investigated by two deletion mutants lacking the substrate-binding domain and linker region, PhaZpst delta sbd and PhaZpstcore. The two deletion mutants had no ability for hydrolysis of water-insoluble P(3HB), while the hydrolysis activities of two deletion mutants for water-soluble 3HB oligomer and its derivatives (dimer, trimer, and tetramer) were identical with those of the wild type, indicating that the function of catalytic domain is independent of its substrate-binding domain and linker region. The hydrolyzed products analysis of 3HB oligomers by HPLC showed that the active site of catalytic domain recognizes at least two 3HB units for hydrolysis. The initial rates of hydrolysis of dimer derivative were lower by 2 orders of magnitude than those of trimer and tetramer derivatives, suggesting that 3HB oligomer derivatives larger than trimer are favorite substrates for PhaZpst.

Carboxylic Ester Hydrolases↗

PubMed2000

Biodegradable poly(ethylene succinate) (PES). 1. Crystal growth kinetics and morphology.

Crystal growth rates of melt-crystallized poly(ethylene succinate) (PES) with a number-average molecular weight Mn of 21,100 and polydispersity of 1.86, respectively, were studied in an extensive temperature range. On the basis of secondary nucleation theory, only one transition between regime III and regime II was found at around 71 degrees C, but no transition of regime II-->I was detected by kinetic analysis. The ratio of nucleation constants KgIII in regime III to KgII in regime II was influenced by the value of activation energy U*, and tended to 2 as predicted by the theory when U* was set as 3688 cal/mol. The morphological changes at spherulitic level and lamellar level in regime II and regime III were examined by using an optical microscope (OM) and atomic force microscope (AFM). Although spherulitic morphologies were found in both regime II and regime III by optical microscopy, further investigations made by following the crystallization process using OM and by checking the lamellar morphology using AFM revealed morphological changes during the crystal growth process and different crystal morphologies in regime II and regime III. Meanwhile by using ultrathin film (approximately 100 nm in thickness), the morphology of lamellar crystals of lozenge-shape outline and composed of single crystals has been studied by AFM technique. The results indicate that there is indeed a transition from regime II to regime III at around 71 degrees C for the isothermal crystallization of PES with an increasing degree of undercooling.

Biodegradation, Environmental↗

PubMed2000

Biodegradable poly(ethylene succinate) (PES). 2. Crystal morphology of melt-crystallized ultrathin film and its change after enzymatic degradation.

Poly(ethylene succinate) (PES) ultrathin films with an initial thickness of approximately 100 nm were prepared by the solution cast method on either cover glass or freshly cleaved mica as the substrate. The ultrathin films were then melt-crystallized at a given temperature for a certain period of time. The surface morphologies of these films on the substrates were observed by an atomic force microscope (AFM) and an optical microscope (OM) under ambient conditions. Two different crystal morphologies having fibril-like structure and flat-on lamellar crystals with a certain width were formed, and their growth mechanisms were discussed in association with previous kinetic data. It has been shown that at a higher crystallization temperature such as 85 degrees C (smaller degree of undercooling) the crystal aggregates tend to form lozenge-shaped hedrites which evolved from a single crystal. The enzymatic degradation of PES crystals on the ultrathin films was carried out by using a PHB depolymerase from Pseudomonas stutzeri at room temperature. The crystal morphologies before and after enzymatic degradation were examined by AFM. The lamellar crystals were hydrolyzed into many small fragments, and these fragments had the same thickness as that of the lamellar crystals before enzymatic degradation. The analysis of morphological results for PES lamellar crystals has revealed the existence of many defects on the surface of melt-crystallized lamellar crystals. These defects were preferentially attacked by the enzyme molecules. Hydrolysis starts from the chains folding in crystal defect area and proceeds along the lateral edges, i.e., along the direction perpendicular to the folding chain.

Biodegradation, Environmental↗

PubMed2000

Microbial degradation of poly(aspartic acid) by two isolated strains of Pedobacter sp. and Sphingomonas sp.

Microbial degradation of thermally synthesized poly(aspartic acid) (PAA) was investigated. A PAA-P1 sample (Mn, 7500; Mw, 20,000; number of branched units/100 monomer units, 3.1) was completely degraded in natural river water within 15 days at 25 degrees C. A new PAA-degrading bacterium (strain KP-2: JCM10638) was isolated together with Sphingomonas sp. KT-1 (JCM10459) from river water, and identified as a member of Pedobacter. A Pedobacter isolate was capable of degrading high-molecular-weight PAA polymers of 5000 to 150,000, and a small amount of low-molecular-weight products of 250 to 5000 was accumulated as residues during the growth of the isolate on PAA. In contrast, the other isolate Sphingomonas sp. KT-1 degraded only low-molecular-weight PAA below 5000. A mixed culturing of Pedobacter sp. KP-2 with Sphingomonas sp. KT-1 resulted in a complete degradation of PAA-P1 sample, but a small amount of low molecular weight components was accumulated during the degradation of highly branched PAA-P2 and PAA-P3 samples.

Bacteria↗

PubMed2000

Biosynthesis and properties of poly(3-hydroxybutyrate-co-3-hydroxyalkanoates) by recombinant strains of Pseudomonas sp. 61-3.

Pseudomonas sp. 61-3 (phbC::tet) strain, which is a phbCPs-disrupted mutant, accumulated a random copolymer consisting of (R)-3-hydroxybutyrate (3HB) and (R)-medium-chain-length 3-hydroxyalkanoate (3HA) units of 6-12 carbon atoms, but the 3HB fraction in the copolymer was less than 50 mol %, resulting in the formation of an amorphus polymer. Therefore, the genes encoding beta-ketothiolase (PhbARe) and NADPH-dependent acetoacetyl-CoA reductase (PhbBRe) from Ralstonia eutropha were expressed under the control of promoters for Pseudomonas sp. 61-3 pha locus or R. eutropha phb operon together with the phaC1Ps gene (PHA synthase 1 gene) from Pseudomonas sp. 61-3 in the phbCPs-disrupted mutant. The recombinant strains synthesized P(3HB-co-3HA) copolymers with very high 3HB compositions (up to 94 mol %) from glucose. The number-average molecular weights of P(3HB-co-3HA) were in the range of 349 x 10(3) to 605 x 10(3). The structure and physical properties of P(3HB-co-3HA) copolymers were characterized by 1H- and 13C-NMR spectrometry, differential scanning calorimetry, and mechanical tensile measurement. P(94% 3HB-co-3HA) copolymer was demonstrated to have good physical properties and to be a flexible material with moderate toughness, similar to low-density polyethylene.

Calorimetry, Differential Scanning↗

PubMed2000

Identification of a marine benthic P(3HB)-degrading bacterium isolate and characterization of its P(3HB) depolymerase.

A poly[(R)-3-hydroxybutyrate] (P(3HB))-degrading marine bacterium (strain NK-1, JCM10458) was isolated from the Pacific Ocean deep-sea floor (1165 m in depth) in Japan. The organism was a motile and Gram negative, aerobic, and rod-shaped bacterium, and its DNA had a guanine-plus-cytosine content of 57.7 mol%. On the basis of several phenotypic characters and a phylogenetic analysis of the gene coding for 16S rRNA, this strain was identified as Marinobacter sp. The strain required sodium salt for growth in the medium and secreted a P(3HB) depolymerase into the supernatant when it was cultivated on (S)-3-hydroxybutyric acid or P(3HB) as the sole carbon source. The P(3HB) depolymerase (PhaZMsp) was purified to homogeneity from the culture supernatant of Marinobacter sp. by hydrophobic and ion exchange column chromatography and showed a molecular mass of 70 kDa. PhaZMsp was stable at temperatures below 37 degrees C and at pH values of 7.5-10.0. The N-terminal amino acid sequences of both the purified enzyme and the truncated one shared high homologies to the N-terminal and internal sequences of Pseudomonas stutzeri depolymerase, respectively. High-performance liquid chromatography analysis revealed that the enzymatic products of P(3HB) yielded monomer, dimer, and trimer of 3-hydroxybutyric acid. PhaZMsp was capable of hydrolyzing P(3HB), poly(3-hydroxypropionate), and poly(4-hydroxybutyrate).

Amino Acid Sequence↗

PubMed2000

Dietary zinc deficiency decreases glutathione S-transferase expression in the rat olfactory epithelium.

Zinc deficiency leads to olfactory and gustatory dysfunction, but little is known about the underlying molecular mechanism of this phenomenon. We examined the effect of dietary zinc deficiency on the rat olfactory epithelium. Immunoreactivities of glutathione S-transferase (GST) mu, neuron-specific enolase (NSE) and proliferating cell nuclear antigen (PCNA), and in situ hybridization of GST mu mRNA in the olfactory epithelia were examined under different dietary zinc intake conditions. Adult male rats were fed a zinc-deficient (ZD) diet (0.5 mg zinc/kg diet), whereas control rats, including pair-fed (PF) and zinc-adequate (ad libitum consumption, AL) groups, were fed a zinc-adequate diet (58 mg zinc/kg diet) for 7 wk. We also examined the effect of zinc replacement (ZR) by subsequently feeding half of the ZD group a zinc-adequate diet for 5 wk after the initial 7-wk deprivation. No significant differences in immunoreactivity for NSE in olfactory epithelial receptor cells or for PCNA in basal cells were noted among groups. Intense GST mu immunoreactivity and hybridization signals were observed in olfactory supporting cells of AL, PF and ZR groups, but very minimal or no such signal was noted in ZD rats. Our findings indicated that zinc deficiency reduces GST mu expression in the supporting cells of rat olfactory epithelia but does not affect receptor cell proliferation or maintenance.

Actins↗

PubMed2000

Effect of mutagenic replacement of the carboxyl terminal amino acid, val124, on the properties and regeneration of bovine pancreatic ribonuclease A.

An important role of C-terminal amino acid residues of bovine pancreatic ribonuclease A (RNase A) in the formation of the three-dimensional structure was previously implied. In this study, we replaced the C-terminal amino acid, Val124, with amino acid residues with different properties by site-directed mutagenesis. The recombinant mutant enzymes were purified and subjected to a refolding study after being converted to a fully reduced and denatured state. There was a significant difference among the mutant enzymes in the rate of recovery of the activity when air oxidation was performed: the rate decreased in the order of V124E, V124L, V124G, V124K, V124A, and V124W. On the other hand, the recovery rates for all the mutant RNase A in the presence of GSH and GSSG were almost the same. The recovered activity of V124E after 24 h incubation reached approximately 90% of that of the wild type enzyme, followed by V124L 80%, V124A and V124W 65%, and V124K and V124G 50%. The duration of the initial lag phase became shorter in the order of V124W, V124A, V124K or V124G, V124E, or V124L. The results imply that the C-terminal amino acid significantly influences the formation of correct disulfide bonds during the refolding process and that the hydrophobic interaction of Val124 is important for efficient packing of the RNase A molecule.

Animals↗

PubMed2000

Differential scanning calorimetry of light meromyosin fragments having various lengths of carp fast skeletal muscle isoforms.

Various recombinant light meromyosin (LMM) fragments were prepared from cDNAs encoding the 10 degrees C and 30 degrees C types of myosin heavy chain isoforms predominantly expressed in fast skeletal muscles of the 10 degrees C- and 30 degrees C-acclimated carp, respectively. These included three kinds of quarter fragments, 1/4-, 2/4-, and 4/4-quarter, composed of residues 1-130, 131-270, and 401-563 from the N-terminus, respectively, as well as three halves, N-, M-, and C-half fragments, containing residues 1-301, 131-400, and 302-563, respectively, and 69K fragments of residues 1-525. Unfortunately, in spite of extensive efforts, the 3/4-quarter fragment was not expressed for both 10 degrees C and 30 degrees C types in our expression system using Escherichia coli. All the LMM fragments except for the 10- and 30-2/4 quarters for the 10 degrees C and 30 degrees C types, respectively, exhibited a typical pattern of a-helix in CD spectrometry. When these were subjected to differential scanning calorimetry (DSC), 30 degrees C-type LMM fragments were all found to be more thermostable than the 10 degrees C-type counterparts. To identify amino acid substitutions responsible for different thermostabilities between the 10 degrees C- and 30 degrees C-type LMMs, six mutant proteins were prepared, mainly focusing on substitutions in the C-terminal half of LMM, and subjected to DSC and CD analyses. For three mutants in which two residues of the 10 degrees C type were replaced by those of the 30 degrees C type, 10-S355T/T361A, 10-M415L/L417V, and 10-S535A/H536Q, the endothermic peaks in DSC increased by 1.4-2.0 degrees C from that of the original 10 degrees C type. The T(m) values for two single-residue substitutions, 10-H449R and 10-T491I, shifted 0.8 and 1.3 degrees C higher than that for the 10 degrees C-type LMM, respectively, whereas the last mutant, 10-G61V, showed no change in thermostability. The finding that the difference in T(m) values for major endothermic peaks from the 10-69K and 30-69K fragments was 4.6 degrees C, which roughly corresponds to that between the original 10 degrees C and 30 degrees C types, suggested that the eight substitutions located in the C-terminal region of the 69K fragments (residues 302-525) are major candidates for the residues responsible for the difference in thermostability between the 10 degrees C- and 30 degrees C-type LMMs.

Amino Acid Sequence↗

PubMed2000

Development of different phenotypes of hypertensive heart failure: systolic versus diastolic failure in Dahl salt-sensitive rats.

OBJECTIVE: There are two phenotypes of heart failure, systolic failure and isolated diastolic heart failure with preserved left ventricular systolic function. Although isolated diastolic heart failure frequently occurs, there are only models for diastolic dysfunction unassociated with heart failure and models with overt diastolic heart failure have not been established. We attempted to develop two different models, i.e. diastolic and systolic failure models, based on hypertension. MATERIALS AND METHODS: Dahl salt-sensitive rats were placed on 8% NaCl diet from 7 weeks old (7-week starting group) or 8 weeks old (8-week starting group). As an age-matched control, Dahl salt-sensitive rats were consistently placed on normal chow. In these rats, echocardiogram was serially recorded, followed by hemodynamic and histological studies. RESULTS: The 7-week starting rats showed a steep elevation in blood pressure and progressive left ventricular hypertrophy, and fell into overt heart failure at approximately 19 weeks. The development of heart failure was not associated with a decrease in left ventricular midwall fractional shortening or an increase in left ventricular end-diastolic dimension as compared with the age-matched control, which mimics the characteristics of clinically observed isolated diastolic heart failure. The 8-week starting rats showed a gradual rise in blood pressure and less progressive left ventricular hypertrophy, and fell into heart failure at approximately 26 weeks with a decrease in mid-wall fractional shortening and an increase in left ventricular end-diastolic dimension. Hemodynamic and histological studies at failing stage revealed comparable elevation of left ventricular end-diastolic pressure and comparable left ventricular fibrosis in both groups. CONCLUSION: These two different models of overt heart failure may be useful as models of isolated diastolic heart failure and systolic heart failure based on the same hypertensive heart disease, respectively, and may contribute to discrimination of the mechanisms of the development of the two different phenotypes of heart failure.

Animals↗

PubMed2000

Internal jugular vein thrombosis, Lemierre's syndrome; oropharyngeal infection with antibiotic and anticoagulation therapy--a case report.

The authors present a case of Lemierre's syndrome that is an uncommon septic thrombophlebitis of the internal jugular vein. A 31-year-old man developed pharyngeal pain one month before hospital admission when he suffered from a severe headache and painful swelling of the left side of his neck. He was diagnosed with tonsillitis. Contrast-enhanced computed tomography and magnetic resonance imaging of the neck revealed the presence of an occlusive thrombosis of the left internal jugular vein and an inflamed mesopharynx. His symptoms and the jugular vein thrombus showed remarkable improvement after administration of antibiotic and anticoagulation therapy. No pulmonary embolism or other metastatic infection were observed. It was suggested that accurate diagnosis during early treatment is essential to obtain a successful prognosis for Lemierre's syndrome.

Adult↗

PubMed2000

Radiofrequency catheter ablation of a concealed atrioventricular accessory pathway associated with a coronary sinus diverticulum.

A 31-year-old woman underwent radiofrequency catheter ablation of a concealed left posteroseptal accessory pathway associated with a coronary sinus diverticulum. The patient had previously undergone unsuccessful catheter ablation of the posteroseptal region of the mitral annulus. Coronary sinus venography revealed the presence of the diverticulum near the ostium. An electrogram in the neck of the diverticulum showed the shortest ventriculoatrial conduction time and a large accessory pathway potential during atrioventricular reciprocating tachycardia. The pathway was successfully ablated within the neck of the diverticulum. The findings in this case underscore the importance of coronary sinus venography before ablation.

Adult↗

PubMed2000

Forensic study of sex determination using PCR on teeth samples.

In this study, sex determination using polymerase chain reaction (PCR) on tooth material was evaluated from the viewpoint of forensic medicine. The sensitivity of PCR for detection of the Y chromosome-specific alphoid repeat sequence and the X chromosome-specific alphoid repeat sequence was 0.5 pg of genomic DNA. Sex could be determined by PCR of DNA extracted from the pulp of 16 freshly extracted permanent teeth and dentine including the surface of the pulp cavity of 6 freshly extracted milk teeth. Sex could be determined using the pulp in all 20 teeth (10 male and 10 female) preserved at room temperature for 22 years. For the pulp of teeth stored in sea water, the sex could be determined in all 8 teeth immersed for 1 week and in 5 of 6 teeth immersed for 4 weeks. In the remaining 1 tooth, in which sex determination based on the pulp failed, the sex could be determined correctly when DNA extracted from the tooth hard tissue was examined. For teeth stored in soil, the sex could be determined accurately in all 8 teeth buried for 1 week, 7 of 8 teeth buried for 4 weeks, and in all 6 teeth buried for 8 weeks. When teeth were heated for 30 min, sex determination from the pulp was possible in all teeth heated to 100, 150, and 200 degrees C, and even in some teeth heated to 250 degrees C. When this method was applied to actual forensic cases, the sex of a mummified body estimated to have been discovered half a year to 1 year after death could be determined readily by examination of the dental pulp. In the skeletons of 2 bodies placed under water for approximately 1 year and approximately 11 years and 7 months, pulp tissues had been dissolved and lost, but sex determination was possible using DNA extracted from hard dental tissues. These results indicate that this method is useful in forensic practices for sex determination based on teeth samples.

Adult↗