PubMed Health⌕ Search

Biomedical subjects

J Devesa

Publications and source records attributed to J Devesa.

At least 19 recordsLinked to original sources
PubMed2001

[Emotional disorders in the working population: an emerging problem in occupational health?].

BACKGROUND: To know the prevalence of emotional disorders in a working population. SUBJECTS AND METHOD: Workers in four companies: construction (A), automobile components (B), electrical and electronic systems (C) and management services (D). The design of the study was cross-sectional with the GHQ-28 questionnaire. RESULTS: The prevalence fluctuated between 8,4% (A) and 24,0% (D). Sex, age, educational level and occupation showed significance differences in some companies. CONCLUSIONS: The results suggest an important prevalence of emotional disorders that likely increase as there will be more firms of services.

Adult↗
PubMed2001

Myostatin regulates cell survival during C2C12 myogenesis.

During the myogenic process in vitro, proliferating myoblasts withdraw irreversible from the cell cycle, acquire an apoptosis-resistant phenotype, and fuse into mature myotubes. The key factor regulating both myocyte cell cycle exit and viability during this transition is the the cyclin-dependent kinase inhibitor p21(cip1). Here we show that the expression of myostatin, a TGF-beta superfamily member known to act as a negative regulator of muscle growth, is upregulated in the course of C2C12 cells myogenesis. We also show that transient transfection of C2C12 myobasts with an expression vector encoding mouse myostatin cDNA efficiently inhibits cell proliferation. Paradoxically, myostatin cDNA overexpression also enhances the survival of differentiating C2C12 myocytes, probably by a mechanism involving, at least in part, upregulation of p21(cip1) mRNA. Our results suggest that myostatin role in myogenesis is more complex than initially suggested and involves another level of regulation apart from inhibition of myoblast proliferation.

Animals↗
PubMed2000

Role of growth hormone receptor in HL-60 cell survival.

Although it is presently well established that locally produced growth hormone (GH) plays a major role in the regulation of survival mechanisms in hemopoietic cells, the responsible mechanisms are poorly understood, and the involvement of the GH receptor (GHR) has not even been demonstrated to date. In this work we investigated the presence of GHR in the human promyelocytic leukemia cell line HL-60, as well as the ability of GH treatment to stimulate both GHR and survival signaling pathways downstream GHR. Our results demonstrate that (1) both GHR mRNA and GHR immunoreactivity are present in HL-60 cells; (2) GH treatment results in an increase in the phosphorylation of the GHR-associated Jak2 and Stat3 proteins, indicating the ability of the hormone to induce receptor activation; and (3) activation of GHR increases the activity of Akt, a serine/threonine kinase that plays a prominent role in the regulation of cell survival. Taken together, these results demonstrate that GHR activation promotes survival of HL-60 cells, thus suggesting that GH plays a major role in the regulation of cell survival in the hemopoietic system, via an autocrine/paracrine mechanism.

Animals↗
PubMed2000

Proteolytic processing of human growth hormone (GH) by rat tissues in vitro: influence of sex and age.

Although a wealth of evidence exists indicating that proteolytic cleavage can enhance the biological activity of the growth hormone (GH) molecule, the mechanisms responsible for the generation of GH fragments are not completely understood. In the present work we investigated the ability of different rat tissues to cleave 22 kDa GH, as well as the influence of sex and age, the two major physiological regulators of GH secretion on this process. Our results show that tissue homogenates obtained from rat liver, skeletal muscle or adipose tissue (three well-documented target organs for the hormone) are able to cleave 22K-GH, while the hormone is resistant to cleavage by rat brain homogenates. This process is rather selective for 22K-GH, since the 20 kDa GH variant exhibits stability to degradation by all tissue homogenates investigated. Moreover, only a minor fraction of 22 kDa GH is cleaved under our experimental conditions, suggesting that GH microheterogeneity within the 22 kDa range may also determine hormone susceptibility. Finally, we also found that 22K-GH processing shows important age-related changes (the greatest intensity observed in 4-day-old pups), while no gender-related differences exist in any of the tissues investigated.

Adipose Tissue↗
PubMed2000

N-glycosylated variants of growth hormone in human pituitary extracts.

This study was designed to investigate the existence, in human pituitary extracts, of growth hormone (GH) variants not encoded by the hGH-N gene. Using anion exchange-fast protein liquid chromatography followed by SDS-PAGE, we isolated several basic forms of pituitary GH. Incubation of these basic forms with endoglycosidase F/N-glycosidase F revealed that two of them (about 34 and 12 kD) were N-glycosylated. In contrast, no changes were found when samples were incubated with the O-linked glycosylation-specific O-glycosidase. Since the GH-N molecule lacks consensus sequences for N-linked glycosylation, our findings suggest that GH genes other than hGH-N are expressed in the human pituitary gland.

Amino Acid Sequence↗
PubMed1999

Activation of growth hormone receptor delivers an antiapoptotic signal: evidence for a role of Akt in this pathway.

A signaling pathway was delineated by which GH promotes cell survival. Experiments were performed in human leukemic cells (HL-60) and Chinese hamster ovary (CHO) cells. In HL-60 cells, GH treatment reduced starvation-induced cell death. In contrast, when HL-60 cells were treated with an anti-GH antibody, cell survival was sharply reduced. In CHO cells stably expressing either the wild-type (wtGHR) or a truncated form (delta454GHR) of the GH receptor in which GH induces a sustained activation of the receptor-associated tyrosine kinase JAK2, we found that GH stimulation inhibited programmed cell death induced by withdrawal of survival factors. This effect was enhanced in cells expressing the truncated form. In contrast, GH did not affect cell survival in CHO cells transfected with either the empty vector or a mutated GHR unable to transduce the signal (4P/AGHR). We also showed that the inhibitory action of GH on apoptosis is probably mediated via stimulation of the serine-threonine kinase Akt, as 1) GH treatment induces a prompt phosphorylation of Akt; and 2) GH effects on cell survival are abolished by transfection of an Akt mutant that exhibits dominant negative function. Experiments with pharmacological inhibitors demonstrated that GH-induced Akt phosphorylation is dependent on phosphoinositide 3-kinase activation. In contrast, we found no changes in Bcl-2 levels secondary to GHR activation.

Animals↗
PubMed1998

Correlation of Pit-1 gene expression and Pit-1 content with proliferation and differentiation in human myeloid leukemic cells.

The transcription factor pituitary-1 (Pit-1) is a homeodomain-containing protein that is expressed mainly in the pituitary, where it drives the expression of growth hormone, prolactin, and thyroid-stimulating hormone beta chain genes. In addition, Pit-1 is required for adequate pituitary cell growth and may be involved in the pathogenesis of pituitary adenomas. Pit-1 expression has been also reported in nonpituitary tissues, where it might be involved in the control of cell proliferation. In order to elucidate such a possibility, we have investigated the changes in both Pit-1 mRNA and Pit-1 immunoreactivity in HL-60 cells following the addition of several differentiating agents. Our results show that while high Pit-1 levels are found in exponentially growing HL-60 cells, a significant decrease occurs after induction of cells to differentiate along the macrophage lineage with 12-O-tetradecanoylphorbol-13-acetate (TPA). In contrast no changes were observed when cells were treated with interferon-alpha, which also induces differentiation of HL-60 cells that, at odds with TPA, is not accompanied with growth arrest. In all, these findings suggest that Pit-1 expression is specifically associated with proliferation in HL-60 cells, thus supporting the idea that one of the functions of nonpituitary Pit-1 may be the control of cell proliferation.

Cell Differentiation↗
PubMed1996

Expression of the human growth hormone normal gene (hGH-N) in proliferating and differentiated HL-60 cells.

The possibility that human leukemic cells could synthesize growth hormone (GH) was investigated in the HL-60 cell line. Western blot analysis of protein extracts obtained from these cells revealed the existence of a major immunoreactive GH (irGH) band, with an approximate molecular weight of 22 kDa, together with lower amounts of 20- and 44-kDa bands. Stimulating proliferating HL-60 cells with KCl clearly increased GH concentration in the incubation medium as compared to basal values. RT-PCR amplification of HL-60 RNA and restriction assay of the amplimers demonstrated that those proteins were the result of the expression of the GH-N (normal) gene in this cell line. These results were confirmed by Northern blot, which also showed that the rate of GH-N gene expression was clearly dependent upon the proliferative state of the cells: while GH transcripts were easily detectable in actively proliferating cells, only minute amounts were observed when cells were induced to differentiate with dimethyl sulphoxide (DMSO). Similar differences were observed by Western blot. In all, these findings demonstrate that HL-60 cells are capable to produce and secrete a GH identical to pituitary GH. Interestingly, the rate of synthesis of the hormone dramatically increases when cells are actively proliferating. Therefore, it is likely that locally produced GH might be involved in the control of leukemic cell proliferation. Further studies are now in course to establish whether this mechanism occurs via an autocrine and/or paracrine way.

Base Sequence↗
PubMed1995

Regulation of hypothalamic somatostatin by glucocorticoids.

Glucocorticoids (GCs) play a key role in the physiology of the hypothalamic-somatotroph axis, since these steroids enhance growth hormone (GH) gene transcription and increase GHRH receptor synthesis. However, GC excess inhibits normal growth in all species studied. This is mainly due to the impaired GH secretion observed during hypercortisolism, a situation in which GH responses to a number of stimuli, including GHRH, are blunted. The inhibitory effect of GCs on GH secretion seems to be dependent on enhanced hypothalamic SS secretion. Since SS release is stimulated by beta-adrenergic agonism we tested the possibility that GC inhibition of GH secretion would depend on increased beta-adrenoceptor activity in SS-producing neurons. The experimental design consisted in evaluating the GH response to GHRH in normal subjects after having induced hypercortisolism, with DEX, and blocking beta-adrenoceptors with propranolol (PRO). Moreover, to investigate the specificity of this mechanism, GHRH-induced GH release was tested after inducing hypercortisolism and enhancing alpha 2-adrenergic or muscarinic cholinergic tone, by giving clonidine (CLO) or pyridostigmine (PD), respectively. As expected, nocturnal DEX administration inhibited the GH response to GHRH. In this situation of hypercortisolism, both PRO and CLO, but not PD, were able to reverse the inhibitory effect of DEX on GHRH-elicited release. However, the potentiating effect of these drugs on the GHRH-induced GH secretion was only observed for PRO. These data confirm that GC excess inhibits GH release by increasing hypothalamic SS secretion, and that the mechanism is mediated by GC-induced enhanced beta-adrenergic responsiveness. Therefore, the defective GHRH secretion observed in chronic hypercortisolism must be a consequence of the continuous blockade that SS excess exerts on GHRH-producing neurons. Our postulate agrees with other data in the literature showing that GCs modulate the secretion of some hypothalamic peptides by changing the responsiveness of the producing neurons from alpha 2-adrenoceptors to that of beta-adrenoceptors.

Clonidine↗
PubMed1995

Clonidine potentiates the growth hormone response to a growth hormone releasing hormone challenge in hypothalamic growth hormone releasing hormone deficient rats.

This study was designed to further investigate our postulate regarding the inhibitory role played by central alpha 2-adrenergic pathways on hypothalamic somatostatin (SS) release in rats. The growth hormone (GH) responses to exogenous GH-releasing factor (GRF; 3 micrograms/kg i.v.) or clonidine (CLO; 100 micrograms/kg i.v.), either given alone or in combination, were tested in 3-month-old male rats made GH-releasing hormone (GH-RH) deficient neonatally by administration of monosodium glutamate (MSG; 4 mg/g body weight s.c.). To prevent the presumable decrease in the pituitary GH content in these animals from leading to an erroneous interpretation of the results obtained, half of these rats were given GRF (MSG-GRF rats; 30 micrograms/kg s.c.) for 3 days immediately prior to GH testing. The other half of MSG-treated and non MSG-treated rats received saline during these days (MSG-S and controls, respectively). To establish the efficiency of GRF priming, the pituitary GH content was measured in other MSG-GRF, MSG-S, and control animals. The mean (+/- SEM) GH peaks in response to GRF challenge were significantly higher in controls than in MSG-GRF rats (125.2 +/- 28.5 vs. 67.5 +/- 19.4 micrograms/l; p < 0.05), while no significant GRF-induced GH release was observed in the MSG-S group. Most likely these results are related to the different pituitary GH content, significantly (p < 0.01) higher in controls than in MSG-GRF rats, and in the latter higher than in MSG-S animals (p < 0.05). CLO administration did not evoke a significant GH release in MSG rats, whether primed with GRF or not.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Studies on alpha 2-adrenergic modulation of hypothalamic somatostatin secretion in rats.

This study was undertaken to investigate whether or not alpha 2-adrenergic pathways would negatively modulate the hypothalamic somatostatin release in rats. To induce pharmacological changes in SS release, three groups of male Sprague-Dawley rats (n = 30/group) were separately anesthetized by ip administration of urethan (which increases SS tone; 1.2 g/kg), pentothal (which impairs SS release; 30 mg/kg), or ketamine (which does not affect spontaneous SS secretion; 40 mg/kg). Ten rats from each group were challenged with GRF (2 micrograms/kg iv), clonidine (40 micrograms/kg iv), or GRF plus clonidine. Administration of clonidine markedly increased the GH responsiveness to GRF in rats anesthetized with urethane or ketamine. In contrast, the GH response to GRF was not modified by clonidine in rats anesthetized with pentothal. These results show that alpha 2-adrenergic stimulation only modifies the GRF-induced GH rise when SS release is high. Therefore, in rats, central alpha 2-adrenergic pathways appear to play a main inhibitory effect on hypothalamic SS secretion.

Animals↗
PubMed1993

Clonidine potentiates the growth hormone (GH) response to GH-releasing hormone in norepinephrine synthesis-inhibited rats: evidence for an alpha-2-adrenergic control of hypothalamic release of somatostatin.

The aim of this study was to investigate whether central alpha 2-adrenergic pathways act, in rats, by inhibiting somatostatin (SS) release, as it has been postulated to occur in other species. The growth hormone (GH) responses to GH-releasing hormone (GRF, 3 micrograms/kg i.v.) or clonidine (CLO, 100 micrograms/kg, i.v.), either given alone or in combination, were tested in male rats in which norepinephrine synthesis had been previously blocked with the dopamine-beta-hydroxylase inhibitor diethyldithio-carbamate (DDTC, 400 mg/kg i.p.). These experiments were also carried out in a control group of animals that had been given placebo (P) instead of DDTC. The GH responses to GRF in the presence of anti-SS serum given to other P and DDTC rats, allowed us to assess whether DDTC treatment had induced increased SS secretion. GH tests were carried out during spontaneous (P) or pharmacologically induced (DDTC) trough periods. Therefore, the mean (+/- SEM) GRF-induced GH peak was similarly low in both groups of rats (P: 66.5 +/- 16.6 micrograms/l; DDTC: 58 +/- 14 micrograms/l). The administration of anti-SS serum significantly (p < 0.01) increased these responses (P: 413 +/- 22; DDTC: 695 +/- 36; p < 0.01 vs. P). CLO administration elicited a maximal GH peak significantly higher (p < 0.05) in P (20.5 +/- 3) than in DDTC rats (4.1 +/- 2); however, the former was significantly lower than GH responses to GRF.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Glucocorticoids may inhibit growth hormone release by enhancing beta-adrenergic responsiveness in hypothalamic somatostatin neurons.

The aim of this study was to investigate the mechanisms by which glucocorticoids inhibit GH secretion in man. In 10 normal volunteers subjects we compared the pattern of GHRH-induced GH release to that elicited by similar challenge given 60 min after a pretreatment with drugs affecting adrenergic and muscarinic cholinergic neurotransmission, both in basal situations and after having induced hypercortisolism. In a first study (P), synthetic GHRH [GRF-(1-29); 1 microgram/kg, i.v.] was administered 60 min after giving placebo. In other experiments, the administration of propranolol (PRO; 40 mg, orally), or clonidine (CLO; 0.300 mg, orally), or pyridostigmine (PD; 120 mg, orally) was followed by GHRH administration 60 min later. These experiments were repeated after giving a nocturnal dose of dexamethasone (DEX; 8 mg, orally at 2300 h). The administration of DEX significantly (P < 0.05) blunted the GH response to GHRH (peaks: 10.7 +/- 3.9 vs. 20.3 +/- 5.5 micrograms/L; DEX vs. P study, respectively). Conversely, either beta-adrenergic blockade (PRO), or alpha 2-adrenergic agonism (CLO), or the enhancement of muscarinic cholinergic tone (PD) significantly increased the GH response to GHRH (peaks: 43 +/- 4.6, 55.6 +/- 5.6 and 51.2 +/- 7, micrograms/L; PRO, CLO, and PD, respectively; P < 0.01 vs. P study). After nocturnal DEX administration, both PRO and CLO, but not PD, were able to reverse the inhibitory effect of DEX on GHRH-elicited GH release (peaks: 39 +/- 5.5, 25.9 +/- 3.9 and 12.9 +/- 3.1, micrograms/L; DEX + PRO, DEX + CLO, and DEX + PD, respectively). However, whereas the potentiating effect of PRO on GHRH-elicited GH release was still observed under hypercortisolism, it was lacking for both CLO and PD when these drugs were given in this situation. These data suggest that the inhibitory effect of glucocorticoid excess on GH release is due to increased hypothalamic somatostatin secretion which appears to be dependent on DEX-induced enhanced beta-adrenergic responsiveness. Moreover, the data further support a major role of hypothalamic alpha 2-adrenergic and beta-adrenergic activities in GH neuroregulation in man.

Adult↗
PubMed1993

A 12-kilodalton N-glycosylated growth hormone-related peptide is present in human pituitary extracts.

This study was designed to investigate whether N-linked glycosylation could account for the presence of glycosylated GH forms (G-GH) in human pituitary extracts. The study was carried out in commercially available pituitary GH preparations (pitGH). Recombinant GHs obtained from eu- or prokaryotic cells were used as controls. Radioiodinated GHs were incubated in tubes containing Concanavalin-A (Con-A) attached to a Sepharose 4B matrix. Pituitary G-GH forms were selectively displaced from Con-A by adding N-acetyl-D-glucosamine or methyl-alpha-D-mannopyranoside and electrophoresed. Autoradiographies of these gels identified a 12-kilodalton (12K) band as the glycosylated form. Due to the fact that this peptide was partially immunoprecipitated with an anti-GH serum and the absence of detectable PRL in the pitGH extracts, it would indicate that such a glycopeptide was GH related. Endoglycosidase-F treatment of pitGH extracts induced a decrease in the mol wt of that 12K peptide, as indicated by the changes observed in its mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This demonstrated that the sugar moieties were N-linked. When recombinant GHs were assayed by a similar method, no specific binding to Con-A was detected. NH2-terminal amino acid sequence analysis from the 12K band demonstrated that this band was composed by three peptides. Peptide 1 corresponds to the GH-N 102-119 sequence. Interestingly, peptide 2 exhibits GH-V 1-18 sequence, while peptide 3 seems to be a novel GH-related peptide. Taken together, these data suggest that the pituitary G-GH form found in human pituitary extracts is derived not from the "normal" GH-N gene, but, rather, from the GH-V gene or another unidentified gene. The potentially important pathophysiological implications of this finding need to be investigated.

Amino Acid Sequence↗
PubMed1991

The role of sexual steroids in the modulation of growth hormone (GH) secretion in humans.

Sex steroids contribute to modulate GH secretion in man. However, both the exact locus and mechanism by which their actions are exerted still remain not clearly understood. We undertook a number of studies designed to ascertain: (1) whether or not sudden or chronic changes in circulating gonadal steroids may affect GH secretion in normal adults; and (2) the reason(s) for gender-related dimorphic pattern of GH release. The pituitary reserve of GH, as evaluated by means of a GHRH challenge, was similar in women with anorexia nervosa and in normally menstruating women. Estrogenic receptor blockade with tamoxifen (TMX) did not significantly change GHRH-induced GH response in these normal women. Therefore, acute or chronic hypoestrogenism apparently had no important effects at level of somatotrophs. In another group of normal women we tested the possibility that changes in circulating estrogens might induce changes in the hypothalamic-somatotroph rhythm (HSR). GHRH challenges were performed throughout a menstrual cycle, and again after having achieved functional ovarian blockade with a GnRH agonist treatment. Short-term ovarian blockade did not significantly affect the parameters of GH response to GHRH, although it was accompanied by an increase in the number of women in a refractory HSR phase at testing. This suggested a low potentiating effect on the basic pattern of somatostatin (SS) release occurring as a consequence of the decrease in circulating estrogens. In normal men, neither the GH response to GHRH nor the HSR were affected by functional testicular blockade (after GnRH agonist treatment). However, the administration of testosterone enanthate (250 mg) to another group of men increased both the GHRH-induced GH release and the number of subjects in a spontaneous secretory HSR phase at testing; these were reversed by estrogenic receptor blockade with TMS. In another group of normal men, the fraction of GH secreted in pulses (FGHP) during a nocturnal sampling period was significantly decreased by testicular blockade. Other parameters of GH secretion,such as the number of GH pulses and their mean amplitude (A), and the mean plasma GH concentration (MCGH), showed a slight, although not significant, decrease following the lack of androgens. The administration of testosterone enanthate (500 mg) reversed these parameters to values similar to those in the basal study. Interestingly, when tamoxifen was given after testosterone enanthate, A, MCGH and FGHP increased to values significantly higher than in any other experimental condition in that study.(ABSTRACT TRUNCATED AT 400 WORDS)

Androgens↗
PubMed1991

Clonidine pretreatment modifies the growth hormone secretory pattern induced by short-term continuous GRF infusion in normal man.

OBJECTIVE: The aim of this study was to investigate the effect of a single dose of clonidine on the pattern of GH release in response to a 10-hour continuous GRF infusion in normal man. DESIGN: Plasma GH was analysed in samples withdrawn at 20-minute intervals, from 0900 to 1900 h, according to the following protocols: in a control study, a placebo was given at 1000 h; in other experiments, clonidine (300 micrograms, orally) was given at 1000 h, alone or together with a continuous intravenous infusion of GRF 1-29 (0.3 micrograms/kg/h) starting at this time. In another experiment, the continuous infusion of GRF 1-29 was preceded by placebo administration at 1000 h. PATIENTS: Eight normal volunteers (four women and four men), aged 19-24 years were studied. MEASUREMENTS: Plasma GH levels were measured by RIA. Analysis of the pattern of GH secretion was performed using cluster analysis. RESULTS: Clonidine induced a slight but significant increase in plasma GH values, peaking 60 to 120 minutes later; however, no significant changes were observed in indices of total and pulsatile GH release for the whole sampling period in this study. Continuous GRF administration led to increased episodic GH secretion, by augmenting GH peak amplitude, although peak frequency was not modified. An increase in interpulse GH values was also observed during GRF infusion. Pretreatment with clonidine clearly changed the pattern of GH release during GRF infusion: the amount of GH secreted was significantly higher, the number of GH peaks significantly increased, and almost all the GH was secreted within them. CONCLUSIONS: These data concord with our previous demonstration that clonidine disrupts the hypothalamic-somatotroph rhythm by inhibiting the hypothalamic release of somatostatin. Given that clonidine pretreatment induced a more physiological episodic pattern of GRF-induced GH release, the possibility of combining clonidine and GRF therapy for short stature in children is envisaged.

Adult↗
PubMed1991

Role of central dopaminergic pathways in the neural control of growth hormone secretion in normal men: studies with metoclopramide.

The aim of this study was to gain further insight into the role that central dopaminergic pathways play in GH neuroregulation in man. Our experimental hypothesis was based on the possibility that most of the controversies on DA role could be due to the fact that the hypothalamic somatotroph rhythm (HSR) was not taken into account when interpreting the GH responses after pharmacological manipulations on dopaminergic pathways. In 10 normal subjects we monitored the effect of central dopaminergic blockade, achieved with metoclopramide (MCP; 10 mg, i.v. bolus), on the pattern of spontaneous GH secretion and the GH responses to a GHRH challenge (GRF1-29, 1 microgram/kg, i.v. bolus) administered together with MCP or 60 min after this drug was given. The study of HSR was made according to our previous postulate. Our results indicate that MCP administration, either prior to or together with the GHRH bolus, significantly increased GHRH-induced GH release during a refractory HSR phase; but not when the GHRH challenge took place during a spontaneous secretory phase. The strong relationship between pre-GHRH plasma GH values and GHRH-elicited GH peaks was lost when MCP was given. These data indicate that MCP was able to disrupt the intrinsic HSR by inhibiting the hypothalamic release of somatostatin (SS). While a main conclusion would be that central DA is a secretagogue for SS secretion, our results also suggest that this role could be dependent on its effects on the adrenergic inputs to SS neurons.

Adult↗