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Dynamics of exogenous nitrogen partitioning and nitrogen remobilization from vegetative organs in pea revealed by 15N in vivo labeling throughout seed filling.

The fluxes of (1) exogenous nitrogen (N) assimilation and (2) remobilization of endogenous N from vegetative plant compartments were measured by 15N labeling during the seed-filling period in pea (Pisum sativum L. cv Cameor), to better understand the mechanism of N remobilization. While the majority (86%) of exogenous N was allocated to the vegetative organs before the beginning of seed filling, this fraction decreased to 45% at the onset of seed filling, the remainder being directed to seeds. Nitrogen remobilization from vegetative parts contributed to 71% of the total N in mature seeds borne on the first two nodes (first stratum). The contribution of remobilized N to total seed N varied, with the highest proportion at the beginning of filling; it was independent of the developmental stage of each stratum of seeds, suggesting that remobilized N forms a unique pool, managed at the whole-plant level and supplied to all filling seeds whatever their position on the plant. Once seed filling starts, N is remobilized from all vegetative organs: 30% of the total N accumulated in seeds was remobilized from leaves, 20% from pod walls, 11% from roots, and 10% from stems. The rate of N remobilization was maximal when seeds of all the different strata were filling, consistent with regulation according to the N demand of seeds. At later stages of seed filling, the rate of remobilization decreases and may become controlled by the amount of residual N in vegetative tissues.

Kinetics↗

Flower Formation in Excised Tobacco Stem Segments: III. Deoxyribonucleic Acid Content in Stem Tissue of Vegetative and Flowering Tobacco Plants.

A method has been developed that extracts DNA from stem tissue of flowering tobacco plants, Nicotiana tabacum cv. Wis. 38. The DNA content of stem tissue from a flowering tobacco plant is correlated with its capacity to flower in vitro. Stem segments known to form 100% floral buds contain 10 times more DNA per gram fresh weight than segments that form 5% floral buds and 95% vegetative buds, and in the uppermost 28 centimeters of flowering tobacco plant stems the DNA content decreases roughly in parallel with the floral gradient.((3)H)-Thymidine incorporation into DNA in the internodes is inhibited by attached leaves in flowering tobacco plants but promoted in vegetative plants. Low indoleacetic acid concentrations (2.8 and 5.7 mum) inhibit DNA synthesis in internodes of defoliated stems of flowering tobacco plants to the same extent as attached leaves, whereas the same concentrations of indoleacetic acid promote DNA synthesis in internodes of defoliated stems of vegetative plants. The optimal concentration (11.4 mum) of indoleacetic acid supplied to defoliated stems of vegetative plants increased the rate of DNA synthesis to 4 times the rate in defoliated stems without indoleacetic acid treatment. This increase more than compensates for the promotion of DNA synthesis by the young leaves. Thus, the opposite effects of young leaves on DNA synthesis in internodes of flowering and vegetative tobacco plants may be related to their auxin content.

Journal Article↗

kurkku, a Phenotype of Acetabularia acetabulum That Is Arrested in Vegetative Growth, Can Be Rescued with Wild-Type Cytoplasm.

We isolated several spontaneous phenotypes in the giant unicell Acetabularia acetabulum that have vegetative terminal morphologies. Because they arrest in vegetative development, these cell lines are effectively immortalized. However, they had to be rescued before they could be studied via classical genetics because no heterozygotes from the original self-crosses were found, that is, the wild-type siblings yielded only wild-type progeny. We attempted to rescue these phenotypes in three ways: by amputating the cell apex, by "piggybacking" the mutant nucleus through development in a binucleate heterokaryon, and by replacing the abnormal apex with a wild-type apex. We used one of our immortal cell lines, kurkku, which has a terminal phenotype consistent with arrest early in the juvenile phase of vegetative development, as a prototype for these rescue methods. The kurkku phenotype segregated 1:3 in the original self-cross in which it arose as if it were a single, recessive Mendelian trait. Although amputation failed to rescue kurkku, we succeeded in compensating for the defect both in binucleate heterokaryons and in apical grafts to wild-type cells. kurkku was always recovered in the progeny of the self-crosses of these grafts. These unique ways of analyzing vegetative mutants, combined with the ability to then perform classical genetics, may make A. acetabulum a powerful unicellular model system for the study of vegetative phase change in plants.

Journal Article↗

Survival of Bacillus cereus spores and vegetative cells in acid media simulating human stomach.

AIMS: To determine the fate of Bacillus cereus spores or vegetative cells in simulated gastric medium. METHODS AND RESULTS: The effects of acidity on the survival of B. cereus in a medium simulating human stomach content was followed on spores at pH 1.0-5.2, and on vegetative cells at pH 2.5-5.7. Gastric media (GM) were prepared by mixing equal volumes of a gastric electrolyte solution with J broth (JB), half-skim milk, pea soup and chicken. At pH 1.0 and 1.4, the number of spores slightly decreased in GM-JB and GM-pea soup and remained stable in GM-milk and GM-chicken. A rapid marked decrease (always higher than 2.0 log CFU ml(-1) in 2 h) in vegetative cell counts was observed at pH below 4.2, 4.0, 3.6 and 3.5 in GM-chicken, GM-JB, GM-milk and GM-pea soup, respectively. Between pH 5.0 and 5.3, B. cereus growth was observed in GM-JB (1.2 log CFU ml(-1) increase after 4 h) and in GM-pea soup (1.8 log CFU ml(-1) increase after 4 h). CONCLUSIONS: Bacillus cereus spores are very much more resistant to gastric acidity than vegetative cells. This resistance strongly depends on the type of food present in the GM. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results suggest that the probability that viable B. cereus cells enter the small intestine, where they can cause diarrhoea, strongly depends on the form of the ingested cells (spores or vegetative cells), on what food they are ingested with, and on the level of stomach acidity.

Bacillus cereus↗

Echocardiographic assessment of vegetations in patients with infective endocarditis: prognostic implications.

Today, echocardiography is the most important technique next to clinical findings and blood cultures in the diagnosis of infective endocarditis. The sensitivity of echocardiography, particularly the transesophageal approach, for detection of vegetations and endocarditis related valvular destructions is high. In addition, echocardiographic findings may have some prognostic implications. The size and mobility of vegetations stratifies endocarditis patients into a high risk group for arterial embolism. In particular, mobile vegetations attached to the mitral valve with a maximal diameter > 10 mm may be prone to embolic events. Furthermore, increase in size of vegetations during antimicrobial treatment may identify patients with no, or at least a prolonged, healing process. Also, a lack of increase in the echo density of vegetations under adequate antibiotic treatment may indicate a poor healing process and may necessitate more aggressive management. The demonstration of paravalvular abscesses by echocardiography, particularly by transesophageal echocardiography, identifies a subgroup of patients who will need urgent cardiac surgery before widespread tissue destruction has occurred.

Echocardiography, Transesophageal↗

Effect of feeding thoroughbred horses a high unsaturated or saturated vegetable oil supplemented diet for 6 months following a 10 month fat acclimation.

This study looked at the effect of feeding diets supplemented with either a predominantly saturated or unsaturated vegetable oil over a prolonged period to exercising horses. Eight Thoroughbred horses were assigned to 2 diet treatments and for 10 months were fed Timothy hay and oats, together with a fortified sweet feed supplemented with either a predominantly unsaturated (Un) or a saturated (S) vegetable oil so that approximately 19% DE (Digestible Energy) came from dietary fat and approximately 12% from either the Un or S source (AC). An increased amount of Un or S fortified sweet feed, replacing the oats, was then fed for a further 6 months (HF) so that approximately 27% DE came from fat and approximately 20% from the Un or S vegetable oil. Standardised incremental treadmill exercise (8-12 m/s) tests (STEP) and duplicate oral glucose tolerance tests (TOL) were carried out after 3, 6 and 9 months of the AC diet and after 3 and 6 months on the HF diet. There was no significant effect of dietary treatment or when the tests were undertaken (time) on the insulin or lactate responses to the STEP tests. Overall there was a significant (P < 0.05) effect of time and treatment on the glucose response, but there was no difference between treatments at the first and last tests or between the results for these tests or between the endAC and endHF tests. No significant effect of treatment or time was seen on the TOL glucose response (% change from Time '0') although there was a trend for the glucose concentrations to be lower and the insulin responses higher (nonsignificant) in the S treatment group. No significant effect of treatment on haematological parameters, monitored monthly, was found. Total protein and gamma glutamyl transferase remained within the normal range throughout. There was a significant effect of treatment (P < 0.05) on cholesterol and triglycerides with higher concentrations in the S group from the first (1 month) sample. Linoleic acid was the main fatty acid in all the 4 plasma lipid classes with slightly, but significant (P < 0.05), higher concentrations in Un for the cholesterol ester and phospholipid classes. There was no effect of time. Overall, the total resting plasma fatty acid content was significantly higher (P < 0.05) with S at the sample points (endAC and endHF). No adverse effects of feeding either diet on apparent coat condition or hoof appearance were seen apart from an apparent increase in the grease score. Many of the parameters assessed showed significant improvements with time (P < 0.05). In conclusion, no apparent adverse effects of feeding a diet supplemented with either an unsaturated or saturated vegetable oil for 6 months at approximately 20% DE after 10 months at approximately 12% DE were identified and there were no apparent disadvantages of feeding a saturated vegetable oil supplemented diet compared with an unsaturated one.

Adaptation, Physiological↗

Enhancement of Interdecadal Climate Variability in the Sahel by Vegetation Interaction.

The role of naturally varying vegetation in influencing the climate variability in the West African Sahel is explored in a coupled atmosphere-land-vegetation model. The Sahel rainfall variability is influenced by sea-surface temperature variations in the oceans. Land-surface feedback is found to increase this variability both on interannual and interdecadal time scales. Interactive vegetation enhances the interdecadal variation substantially but can reduce year-to-year variability because of a phase lag introduced by the relatively slow vegetation adjustment time. Variations in vegetation accompany the changes in rainfall, in particular the multidecadal drying trend from the 1950s to the 1980s.

Journal Article↗

OCCURRENCE OF COLIFORMS, FECAL COLIFORMS, AND STREPTOCOCCI ON VEGETATION AND INSECTS.

This study considers the sanitary significance of coliforms, fecal coliforms, and streptococci isolated from 152 species of plants and 40 samples of insects. These specimens were collected from various ecological environments and grouped into several categories. Results indicate that typical coliforms of the warm-blooded animal gut contribute a relatively small percentage of the organisms associated with vegetation (14.1%) and insects, (14.9%). A total of 1,203 coliform strains from vegetation and 1,084 coliform strains from insects were classified as to IMViC type and fecal coliform. No type was predominant in either the vegetation or insect groupings. The biochemical results for 646 streptococci from vegetation and 226 cultures from insects were reported. The predominant group, Streptococcus fecalis, as defined by Sherman criteria, constituted a majority of all strains from vegetation and insects. The "Completed Coliform Test" is recommended for the examination of plant and insect specimens to eliminate the many anaerobic and aerobic sporeforming bacteria that frequently produce false positive reactions by the "Confirmed Test" procedure. These findings support the current interpretation of the significance of the fecal coliform test for stream investigations or for surface water quality evaluations.

Animals↗

Pathogenesis of Clostridium botulinum Type A: Study of In Vivo Toxin Release by Implantation of Diffusion Chambers Containing Spores, Vegetative Cells, and Free Toxin.

Millipore diffusion chambers (MDC) with 0.22-mum filters loaded with spores or vegetative cells of Clostridium botulinum were surgically implanted intraperitoneally (ip) into guinea pigs. MDC expose C. botulinum spores or vegetative cells to body fluids yet protect them from phagocytes. Guinea pigs receiving MDC containing 10(9) spores plus 10(8) polymorphonuclear (PMN) leukocytes and MDC with 10(9) vegetative cells died within 48 hr, indicating that toxin was released and diffused out. MDC with 10(9) spores alone allowed 90% of animals to survive for at least 96 hr. Microscopically, it was observed that vegetative cells in MDC were disintegrated and leukocytes plus spores were phagocytized and germinated; spores alone remained intact and phase bright. Chemotactic attraction of leukocytes to MDC walls was also observed. Apparently, body fluids do not attack spores; thus, PMN leukocyte engulfment is essential for germination and release of spore-bound toxin in this type of C. botulinum pathogenesis. However, vegetative cells appear to be attacked by bacteriolytic enzymes (e.g., lysozyme) in body fluids, and leukocyte engulfment is not essential for toxin release.

Journal Article↗

Epsilon-toxin is required for most Clostridium perfringens type D vegetative culture supernatants to cause lethality in the mouse intravenous injection model.

Clostridium perfringens type D enterotoxemias have significant economic impact by causing rapid death of several domestic animal species. Consequently, domestic animals are commonly vaccinated, at varying efficacy, with inactivated type D vegetative supernatants. Improved type D vaccines might become possible if the lethal toxins produced by type D isolates were characterized and the contributions of those toxins to supernatant-induced lethality were established. Therefore, the current study evaluated the presence of lethal toxins in supernatants prepared from late-log-phase vegetative cultures of a large collection of genotype D isolates. Under this growth condition, most genotype D isolates produced variable levels of at least three different lethal toxins, including epsilon-toxin (ETX). To model the rapid lethality of type D enterotoxemias, studies were conducted involving intravenous (i.v.) injection of genotype D vegetative supernatants into mice, which were then observed for neurotoxic distress. Those experiments demonstrated a correlation between ETX (but not alpha-toxin or perfringolysin O) levels in late-log-phase genotype D supernatants and lethality. Consistent with the known proteolytic activation requirement for ETX toxicity, trypsin pretreatment was required for, or substantially increased, the lethality of nearly all of the tested genotype D vegetative supernatants. Finally, the lethality of these trypsin-pretreated genotype D supernatants could be completely neutralized by an ETX-specific monoclonal antibody but not by an alpha-toxin-specific monoclonal antibody. Collectively, these results indicate that, under the experimental conditions used in the present study, ETX is necessary for the lethal properties of most genotype D vegetative supernatants in the mouse i.v. injection model.

Animals↗

Properties and developmental roles of the lysyl- and tryptophanyl-transfer ribonucleic acid synthetases of Bacillus subtilis: common genetic origin of the corresponding spore and vegetative enzymes.

The lysyl-transfer ribonucleic acid synthetase (LRS) and tryptophanyl-transfer ribonucleic acid synthetases (TRS) (l-lysine:tRNA ligase [AMP], EC 6.1.1.6; and l-tryptophan:tRNA ligase [AMP], EC 6.1.1.2) have been purified 60- and 100-fold, respectively, from vegetative cells and spores of Bacillus subtilis. There are no significant differences between the corresponding spore and vegetative enzymes with respect to their elution characteristics from columns of phosphocellulose or hydroxylapatite, their molecular weight (~130,000 for LRS and ~87,000 for TRS as determined by gel filtration), their kinetic constants for substrates (in the amino acid-dependent adenosine triphosphate-pyrophosphate exchange reaction), and the kinetics of inactivation by heat and by antibody. The Mg(2+) requirement for optimal enzyme activity of the corresponding spore and vegetative enzyme differ slightly. Mutants having defective (temperature sensitive) vegetative LRS or TRS activities produce spores in which these enzymes are also defective. The mutant spores are more heat sensitive than the parental type, but contain normal levels of dipicolinic acid. They germinate normally at the restrictive temperature (43 C), but are blocked at specific developmental stages in outgrowth. No modification in temperature sensitivity phenotype occurs during outgrowth, nor is there a change in molecular weight of the two enzymes. The implication is that the LRS and TRS activities of the vegetative and spore stages are each coded (at least in part) by the same structural gene. The temperature sensitivity of mutant spores is discussed with respect to those factors which are involved in the formation of the heat-resistant state.

Amino Acyl-tRNA Synthetases↗

The spectrum of spontaneous rifampin resistance mutations in the rpoB gene of Bacillus subtilis 168 spores differs from that of vegetative cells and resembles that of Mycobacterium tuberculosis.

Mutations causing rifampin resistance in vegetative cells of Bacillus subtilis 168 have thus far been mapped to a rather restricted set of alterations at either Q469 or H482 within cluster I of the rpoB gene encoding the beta subunit of RNA polymerase. In this study, we demonstrated that spores of B. subtilis 168 exhibit a spectrum of spontaneous rifampin resistance mutations distinct from that of vegetative cells. In addition to the rpoB mutations Q469K, Q469R, and H482Y previously characterized in vegetative cells, we isolated a new mutation of rpoB, H482R, from vegetative cells. Additional new rifampin resistance mutations arising from spores were detected at A478N and most frequently at S487L. The S487L change is the predominant change found in rpoB mutations sequenced from rifampin-resistant clinical isolates of Mycobacterium tuberculosis. The observations are discussed in terms of the underlying differences of the DNA environment within dormant cells and vegetatively growing cells.

Amino Acid Sequence↗

Localization of the vegetative cell wall hydrolases LytC, LytE, and LytF on the Bacillus subtilis cell surface and stability of these enzymes to cell wall-bound or extracellular proteases.

LytF, LytE, and LytC are vegetative cell wall hydrolases in Bacillus subtilis. Immunofluorescence microscopy showed that an epitope-tagged LytF fusion protein (LytF-3xFLAG) in the wild-type background strain was localized at cell separation sites and one of the cell poles of rod-shaped cells during vegetative growth. However, in a mutant lacking both the cell surface protease WprA and the extracellular protease Epr, the fusion protein was observed at both cell poles in addition to cell separation sites. This suggests that LytF is potentially localized at cell separation sites and both cell poles during vegetative growth and that WprA and Epr are involved in LytF degradation. The localization pattern of LytE-3xFLAG was very similar to that of LytF-3xFLAG during vegetative growth. However, especially in the early vegetative growth phase, there was a remarkable difference between the shape of cells expressing LytE-3xFLAG and the shape of cells expressing LytF-3xFLAG. In the case of LytF-3xFLAG, it seemed that the signals in normal rod-shaped cells were stronger than those in long-chain cells. In contrast, the reverse was found in the case of LytE-3xFLAG. This difference may reflect the dependence on different sigma factors for gene expression. The results support and extend the previous finding that LytF and LytE are cell-separating enzymes. On the other hand, we observed that cells producing LytC-3xFLAG are uniformly coated with the fusion protein after the middle of the exponential growth phase, which supports the suggestion that LytC is a major autolysin that is not associated with cell separation.

Amidohydrolases↗

Biochemistry of sporulation. I. Metabolism of acetate by vegetative and sporulating cells.

Hanson, Richard S. (University of Illinois, Urbana), V. R. Srinivasan, and H. Orin Halvorson. Biochemistry of sporulation. I. Metabolism of acetate by vegetative and sporulating cells. J. Bacteriol. 85:451-460. 1963.-The transition from the vegetative to the sporulating cycle in a sporeformer is marked by a change in the enzymatic machinery of the cell. When vegetative cells of Bacillus cereus strain T are grown in a glucose-yeast extract-minerals medium, acetate accumulates until the beginning of the sporulation cycle. The acetate-activating systems are present in the vegetative cells as well as in the cells of the early stages of sporulation, whereas the enzymes necessary for the terminal oxidation of acetate to carbon dioxide are absent in the vegetative stage. The induction of a functional tricarboxylic acid cycle during early sporulation is inhibited by chloramphenicol. alpha-Picolinic acid also prevents morphological, as well as physiological, changes during the transition.

Acetates↗

LETHALITY FOR MICE OF VEGETATIVE AND SPORE FORMS OF BACILLUS CEREUS AND BACILLUS CEREUS-LIKE INSECT PATHOGENS INJECTED INTRAPERITONEALLY AND SUBCUTANEOUSLY.

Lamanna, Carl (Army Research Office, Washington, D.C.) and Linda Jones. Lethality for mice of vegetative and spore forms of Bacillus cereus and Bacillus cereus-like insect pathogens injected intraperitoneally and subcutaneously. J. Bacteriol. 85:532-535. 1963.-Lethality for mice of Bacillus thuringiensis var. thuringiensis (two strains), var. sotto (one strain), var. alesti (one strain), and B. entomocidus var. entomocidus (one strain) maintained on artificial laboratory media is dependent upon the route of injection, the number of the bacilli, and their biological state, i.e., vegetative or spore form. Deaths did not occur upon subcutaneous injection of tenfold greater numbers of organisms than the ld(50) dose for the intraperitoneal route. From 10 to 100 times the number of spores were required to kill than vegetative bacilli given intraperitoneally. These observations duplicated those made with strains of B. cereus. For both types of spore forms, from 5 x 10(5) to 5 x 10(7) vegetative organisms were required for one intraperitoneal ld(50). With two strains of highly virulent B. anthracis, ld(50) values were the same for the vegetative and spore states, and for subcutaneous and intraperitoneal injections. Thus, the insect pathogenic sporeforming bacteria tested are more like B. cereus than like B. anthracis in their capacity to infect mice. The similarity to B. cereus extends to the rapidity of death after fatal injection of the insect pathogenic strains. The medical and taxonomic implications of the observations are discussed.

Animals↗

Nutritional requirements for germination, outgrowth, and vegetative growth of putrefactive anaerobe 3679 in a chemically defined medium.

Fujioka, Roger S. (University of Hawaii, Honolulu), and Hilmer A. Frank. Nutritional requirements for germination, outgrowth, and vegetative growth of putrefactive anaerobe 3679 in a chemically defined medium. J. Bacteriol. 92:1515-1520. 1966.-A chemically defined medium was used to study the nutritional requirements for germination, outgrowth, and vegetative growth of putrefactive anaerobe 3679. Vegetative growth required arginine, cysteine, phenylalanine, threonine, valine, tryptophan, K(2)HPO(4), and Na(+); was markedly stimulated by isoleucine, tyrosine, nicotinic acid, and p-aminobenzoic acid; and was stimulated slightly by alanine, biotin, pyridoxamine, glucose, and salts (MgSO(4), FeSO(4), MnSO(4)). Growth occurred over an initial pH range of 6.0 to 8.2, and at incubation temperatures ranging from 20 to 45 C. No autolysis occurred during vegetative growth, although loss of motility and cell settling were observed upon prolonged incubation. Vegetative growth was inhibited completely by aerobic conditions. Completion of spore germination was inhibited at pH 4.8, outgrowth was inhibited at pH 8.5 and higher, and germination was inhibited at 9.0 and higher. Slow germination, but no outgrowth, was observed at 8 C; at 47 and 52 C, spore inocula germinated only partially. Under aerobic conditions, all inoculated spores germinated completely, and one-half of these also emerged. Alanine was considered the primary germinant in the chemically defined medium, and arginine and mineral salts (MgSO(4), FeSO(4), MnSO(4), and NaCl), as secondary germinants. During outgrowth of germinated spores, cysteine and K(+) were required for emergence, Na(+) for elongation, and arginine and valine for division of the elongated cell.

Alanine↗

Properties of fructose 1,6-diphosphate aldolases from spores and vegetative cells of Bacillus cereus.

Fructose 1,6-diphosphate aldolase from cells of Bacillus cereus appears to be typical Class II aldolase as judged by its functional and physical properties. Spore and vegetative cell aldolase had similar enzymatic, immunochemical, and heat resistance properties in the absence of calcium, but they differed in their thermal stabilities in the presence of calcium, their Stokes' radii, their mobility in acrylamide gel electrophoresis, and their molecular weights. The pH optimum for both enzymes was 8.5, and their K(m) with respect to substrate was 2 x 10(-3)m. Highly purified spore and vegetative cell aldolases were both heat labile with half-lives of 4 min at 53 C and pH 6.4. In the presence of 3 x 10(-2)m solution of calcium ions, the stability of the spore protein increased 12-fold but the vegetative form became more heat labile. The enhanced stability of the spore aldolase was not diminished by dialysis or gel filtration but was lost after chromatography on diethylaminoethyl cellulose at pH 7.4. Aldolase from vegetative cells exists in an equilibrium mixture of two molecular weights, 115,000 and 79,000 in the approximate ratio of 1:4, respectively. The molecular weight of spore aldolase is 44,000. Spore aldolase was more mobile during electrophoresis than its vegetative cell counterpart because of its smaller size.

Aldehyde-Lyases↗

Prognostic implication of detecting vegetations by M-mode echocardiography.

The prognostic significance of finding vegetations by M-mode echocardiography in patients with infective endocarditis remains controversial because many such patients are referred for early surgery. We detected vegetations in 7 of 25 consecutive subjects with active infective endocarditis (28.1%) studied by M-mode echocardiography. 11 patients died within 1 year and only 1 underwent surgery. Patients with vegetations had a higher likelihood of developing serious complications such as death, congestive heart failure or "severe congestive heart failure' when compared with patients without vegetation. This study validates previously reported series including a high percentage of patients "requiring surgery' and points to the serious prognostic implication of finding a vegetation by M-mode echocardiography.

Adolescent↗