Search PubMedSearch

SEARCH · Search PubMed

Results for “potentially damaging variation”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 19 recordsLinked to original sources

Phenotypic Impact of Rare Potentially Damaging Copy Number Variation in Obsessive-Compulsive Disorder and Chronic Tic Disorders.

BACKGROUND: Recent studies report an important-and previously underestimated-role of rare variation in risk of obsessive-compulsive disorder (OCD) and chronic tic disorders (CTD). Using data from a large epidemiological study, we evaluate the distribution of potentially damaging copy number variation (pdCNV) in OCD and CTD, examining associations between pdCNV and the phenotypes of probands, including a consideration of early- vs. late-diagnoses. METHOD: The Obsessive-Compulsive Inventory-Revised (OCI-R) questionnaire was used to ascertain psychometric profiles of OCD probands. CNV were identified genome-wide using chromosomal microarray data. RESULTS: For 993 OCD cases, 86 (9%) were identified as pdCNV carriers. The most frequent pdCNV found was at the 16p13.11 region. There was no significant association between pdCNV and the OCI-R total score. However, pdCNV was associated with Obsessing and Checking subscores. There was no significant difference in pdCNV frequency between early- vs. late-diagnosed OCD probands. Of the 217 CTD cases, 18 (8%) were identified as pdCNV carriers. CTD probands with pdCNV were significantly more likely to have co-occurring autism spectrum disorder (ASD). CONCLUSIONS: pdCNV represents part of the risk architecture for OCD and CTD. If replicated, our findings suggest pdCNV impact some OCD symptoms. Genes within the 16p13.11 region are potential OCD risk genes.

Humans

Interindividual variation in the responses of cultured human lymphocytes to exposure from DNA damaging chemical agents: interindividual variation to carcinogen exposure.

Human population variability to standardized doses of N-acetoxy-2-acetylaminofluorene (NA-AAF) and 7, 12-dimethylbenz(a) anthracene (DMBA) was determined in cultured lymphocytes by measuring (a) differential stimulation of unscheduled DNA synthesis after 1 h induction of DNA damage by 10 micrometer NA-AAF, (b) the level of NA-AAF induced chromosome aberrations remaining after 8 h of DNA-repair synthesis, and (c) the level of [3H]DMBA bound to DNA after 18 h incubation of resting lymphocytes in 5 micrometer DMBA. All 3 parameters indicated individual variation to carcinogen exposure and were correlated to the population differences in age, sex, blood pressure and mortality rates. Males always had a greater potential to accumulate DNA-damage than did females regardless of the sampled population. DNA-damage potentials increased with increasing age, blood pressure or mortality rates. There was always proportionally greater DNA-damage potentials in the males than in females. The in vitro response of mature granulocytes to a 10 micrometer NA-AAF dose, as estimated by [3H] thymidine incorporation from unscheduled DNA synthesis, was much lower than lymphocyte response. Nevertheless, individual variations in granulocyte NA-AAF induced unscheduled DNA synthesis paralleled the inter-individual fluctuations observed in the lymphocyte responses to NA-AAF.

9,10-Dimethyl-1,2-benzanthracene

Intrinsic radiosensitivity and PLD repair in osteosarcoma cell lines.

The response to radiation of seven osteosarcoma cell lines was analysed by in vitro colony-forming assay and compared with that of eight human fibroblast strains. The values of D0, the surviving fraction after 2 Gy (S2Gy), and the mean inactivation dose (D) of osteosarcoma cells in log-phase culture were significantly higher than those of fibroblast strains (p less than 0.01). PLD (potentially lethal damage) repair of osteosarcoma cells evaluated in the plateau phase of growth showed great variation for enhancement of survival, although all of the values were maximized within 12 h after irradiation. In the osteosarcoma, intrinsic radiosensitivity in vitro reflected the clinical response to radiation. However, the capacity for PLD repair might not be a good indicator for predicting the results of radiation therapy.

Cell Survival

Exploiting Omic Data to Advance Predictive Ecotoxicology.

Predicting species-specific chemical sensitivity using in silico approaches has the potential to transform environmental risk assessment, conservation, and biomonitoring, while reducing, and ultimately replacing, animal testing. Genomic and transcriptomic data capture extensive sensitivity-relevant variation, including differences in molecular targets, xenobiotic metabolism, and damage mitigation pathways. Large-scale sequencing initiatives therefore offer an unprecedented opportunity to address ecotoxicology's "too many species" problem. Although existing omic-based predictive tools provide proof of concept, they have so far been applied to a narrow set of relatively straightforward prediction scenarios. To achieve broader applicability, current and future tools must be firmly grounded in the diverse molecular mechanisms underlying differential chemical responses. Here, we critically evaluate the emerging field of predicting species sensitivity using molecular variation inferred from omic data. We analyze the strengths and limitations of current omic-based approaches and identify major sequence and ecotoxicological data gaps, as well as critical bioinformatic challenges. We then review the current knowledge of how molecular biology underlies differential chemical sensitivity, outlining research paths to allow the next generation of sensitivity prediction tools to exploit ever expanding omic data.

Ecotoxicology

The use of direct current to cause selective block of large fibres in peripheral nerves.

The effect of direct current on the propagated compound action potential in a cutaneous branch of the radial nerve of the dog was studied and a convenient method for applying electrical current to a nerve is described. Changes in the flow of blocking current, even when these were made slowly during a period of 5-10 sec, caused stimulation of the nerve. The method was satisfactory for producing differential nerve block within the myelinated fibres. It was confirmed that direct current could be used to eliminate selectively the group II/III potentials. When these potentials had been eliminated for a period of 5 min the mean height of the group IV potentials was reduced to 65% of the control value, and their conduction velocity was decreased by 10%. Relatively small variations in current flow caused a marked change in conduction. In the early stages of block, frequent relatively large reductions in current flow were necessary to produce a constant response and it was impossible to cause a stable effect on conduction until the blocking current had been applied for at least 1 min. Because of nerve damage the method is unsuitable for clinical use.

Action Potentials

[Basis and clinical application of the "contingent negative variation" in "evoked response audiometry" (author's transl)].

After repeated stimulation with an indicative stimulus and a second imperative stimulus, a slow negative potential shife, the "Contingent Negative Variation" (CNV), can be recorded in the EEG before the second stimulus. On account of the main features of CNV two qualitative tests can be used in audiometry. "Tone-CNV" is an objective test of perception of a pure-tone. The indicative pure-tone is followed regularly by a second imperative stimulus, a flash of light or a slide. The appearance of CNV is the positive sign of perception, even if the "acoustically evoked potential" (AEP) is difficult to define. Adults and children (5 to 12 years) with normal hearing or suffering from several forms of hearing loss developed a CNV, except those patients with severe brain damage. The advantage of this method is the short testing time and accentuation of AEPs. The test can be also done under sedation. In "Objective Speech Audiometry", a proof of concept discrimination, only words of one concept are combined with a second imperative stimulus. The other group of words of the second concept remains "unreinforced". After correct discrimination all normal subjects developed a CNV before the second stimulus and after the "unreinforced" concept a "Discriminative Positive Variation" (DPV). Thus, this method can be used for clinical purposes. Loss of discrimination, perseveration phenomena and hemispheric differences were observed in aphasic patients.

Acoustic Stimulation

[Objective diagnosis of recruitment by brainstem audiometry (author's transl)].

The differential diagnosis in cochlear damage with brainstem potentials is easily done by looking for normal latencies that coincide with substantial hearing loss. Further validation can now be achieved with methods representing Fowler and SISI test equivalents. Amplitudes of brainstem potential were compared in 28 patients with marked differences of hearing loss in either ear. Amplitudes are balanced at the stimulus intensity corresponding to subjective loudness balance (Fig. 3). Intersubject comparison of input/output function steepness is not advisable because of the greater interindividual variation of amplitudes (Fig. 2). Amplitude modulation by short Gauss-shaped increments (Fig. 1) or by decrements of a continuous tone evokes clear brainstem potentials at thresholds of about 5dB above subjective detection, but it also depends upon stimulus duration or steepness (Fib. 4), frequency (Fig. 5) and the degree of modulation (Fig. 6). With a cochlear hearing loss of more than 40dB, a 1 dB increment evokes potentials for continuous tone intensities higher than 20 dB above subjective thresholds (Fig. 7). This test is mainly useful for children with retarded language development and lacking a stapedius reflex, for adults, who cannot sufficiently cooperate, and where there is a prolonged latency due to middle ear damage or a steep high frequency loss.

Audiometry

Molecular identification of Hymenopteran insects collected by using Malaise traps from Hazarganji Chiltan National Park Quetta, Pakistan.

The order Hymenoptera holds great significance for humans, particularly in tropical and subtropical regions, due to its role as a pollinator of wild and cultivated flowering plants, parasites of destructive insects and honey producers. Despite this importance, limited attention has been given to the genetic diversity and molecular identification of Hymenopteran insects in most protected areas. This study provides insights into the first DNA barcode of Hymenopteran insects collected from Hazarganji Chiltan National Park (HCNP) and contributes to the global reference library of DNA barcodes. A total of 784 insect specimens were collected using Malaise traps, out of which 538 (68.62%) specimens were morphologically identified as Hymenopteran insects. The highest abundance of species of Hymenoptera (133/538, 24.72%) was observed during August and least in November (16/538, 2.97%). Genomic DNA extraction was performed individually from 90/538 (16.73%) morphologically identified specimens using the standard phenol-chloroform method, which were subjected separately to the PCR for their molecular confirmation via the amplification of cytochrome c oxidase subunit 1 (cox1) gene. The BLAST analyses of obtained sequences showed 91.64% to 100% identities with related sequences and clustered phylogenetically with their corresponding sequences that were reported from Australia, Bulgaria, Canada, Finland, Germany, India, Israel, and Pakistan. Additionally, total of 13 barcode index numbers (BINs) were assigned by Barcode of Life Data Systems (BOLD), out of which 12 were un-unique and one was unique (BOLD: AEU1239) which was assigned for Anthidium punctatum. This indicates the potential geographical variation of Hymenopteran population in HCNP. Further comprehensive studies are needed to molecularly confirm the existing insect species in HCNP and evaluate their impacts on the environment, both as beneficial (for example, pollination, honey producers and natural enemies) and detrimental (for example, venomous stings, crop damage, and pathogens transmission).

Humans

Chemometric insights into Lactiplantibacillus plantarum effects on onion (Allium cepa L.) metabolism and antidiabetic activity under cadmium stress.

Cadmium (Cd) is a toxic heavy metal that causes severe physiological damage in plants, inhibiting growth and ultimately reducing crop yield. Lactic acid bacteria regulate Cd availability through bioaccumulation and biosorption. This study evaluated the Cd tolerance of Lactiplantibacillus plantarum 10CH by determining its survival capacity under Cd stress and its potential to mitigate Cd-induced stress in onion (Allium cepa L.). The bacterial strain tolerated Cd concentrations up to 100 µM, and whole-genome sequencing identified genes involved in Cd biosorption, accumulation, and efflux. Exposure of onion to increasing CdCl2 concentrations significantly reduced root and shoot biomass. Inoculation with Lb. plantarum 10CH alleviated Cd stress at 100 µM, enhancing root and shoot biomass, reducing Cd accumulation, lowering oxidative damage markers, and stimulating antioxidant enzyme activities. Metabolic profiling revealed that Cd stress significantly reduced primary metabolites and amino acids, particularly at 100 µM, while bacterial inoculation restored key amino acids and peptides, including arginine, tyrosine, and glutamic acid. Chemometric analysis using unsupervised (PCA) and supervised (OPLS-DA) models revealed clear metabolite variation among untreated, Cd-stressed, and bacterial inoculated Cd-stressed onion leaves. Furthermore, leaf extracts exhibited α-glucosidase inhibitory activity, with the highest activity in control plants (IC50 = 425.2 ± 0.5 µg/mL). Cd-stressed plants showed moderate antidiabetic activity, which was significantly reduced by bacterial inoculation. Overall, these findings demonstrate that Lb. plantarum 10CH can survive under Cd stress and alleviates Cd-induced stress in onion, highlighting its potential as a bioinoculant to mitigate heavy metal stress.

Onions

Flow cytometry of mammalian sperm: progress in DNA and morphology measurement.

Variability in DNA content and head shape of mammalian sperm are potentially useful markers for flow cytometric monitoring of genetic damage in spermatogenic cells. The high refractive index and extreme flatness of the sperm heads produce an optical effect which interferes with DNA measurements in flow cytometers which have dye excitation and fluorescence light collection normal to the axis of flow. Orientation of sperm in flow controls this effect and results in coefficients of variation of 2.5% and 4.2%, respectively, for DNA measurements of mouse and human sperm. Alternatively, the optical effect can be used to generate shape-related information. Measurements on randomly oriented sperm from three mammalian species using a pair of fluorescence detectors indicate that large shape differences are detectable. Acriflavine-Feulgen stained sperm nuclei are significantly bleached during flow cytometric measurements at power levels routinely used in many flow cytometers. Dual beam studies of this phenomenon indicate it may be useful in detecting abnormally shaped sperm.

Animals

Low-pass whole-genome sequencing reveals genomic diversity and ecotype-specific adaptation in indigenous Tigrayan chickens.

Indigenous chickens play a critical role in food security and climate resilience in smallholder systems, yet their genomic diversity and adaptive potential remain insufficiently characterised. This study employed low-pass whole-genome sequencing (LP-WGS; 0.2-1.99×) to investigate genomic diversity, population structure, inbreeding and candidate environment-associated genomic variation in 33 chickens from highland, midland, and lowland agroecologies in the Tigray region of northern Ethiopia. After imputation and stringent filtering, 23.4 million high-confidence SNPs were retained, including ~ 17% novel variants, indicating substantial uncharacterised genetic diversity in these populations. SNP density (13.8 ± 8.6 SNPs/kb) was comparable to values reported from high-coverage Ethiopian chicken datasets, demonstrating the suitability of LP-WGS for population genomics in resource-limited settings. Marked differences in genomic diversity were observed among ecotypes: midland chickens showed the highest nucleotide diversity (π = 0.00267), followed by lowland (π = 0.00233), whereas highland chickens showed the lowest diversity (π = 0.00203) and elevated genomic inbreeding (FROH and FHOM ≈ 0.18). Population structure analyses revealed clear genetic separation among ecotypes. PCA (13.91% variation explained) distinguished lowland chickens along PC1 and separated highland from midland along PC2, while ADMIXTURE and FST patterns supported three major ancestral genomic backgrounds. Functional annotation of private missense variants uncovered distinct adaptive signatures reflecting the contrasting agroecological conditions. Highland chickens showed enrichment of candidate genes potentially involved in physiological processes relevant to high-altitude environments, including cold response, angiogenesis, cardiovascular regulation and metabolic homeostasis (eg., PARP1, ACOX2, ITGB3, EDNRB, SOX8, and SOX10). Midland chickens exhibited candidate signals of selection in genes with known roles in innate antiviral immunity, bacterial defence and inflammatory regulation (eg., BAK1, CLSTN1, CYSLTR1, CYSLTR2, CXCR7, GIPR, DSCAM, GDAP1, TLR3, TLR4, TLR7, IFIH1, ADORA1, EPHB1, and TMPRSS2). Lowland chickens displayed candidate variants associated with heat-stress response, DNA damage repair, oxidative balance and cardiovascular support under extreme temperatures (e.g., MLH1, BDKRB1, GPR19, FLT1, CCL18, TGM2, and RAMP3). Overall, the results indicate substantial genomic differentiation among ecotypes and suggest candidate environment-associated genetic divergence across Tigray's diverse agroecological zones. These populations may represent important reservoirs of adaptive genetic variation for climate-resilient poultry breeding, warranting further functional validation and conservation-oriented management.

Animals

The perineuronal glial reaction after axotomy.

The perineuronal glial reaction after axotomy of the facial nerve was examined in adult mice and rats and in newborn rabbits. The facial nerve was damaged in two ways: by crush lesions, which were followed by complete neuronal regeneration, and by transection (in mice) or evulsion (in rats and rabbits), which caused extensive nerve cell disintegration. After nerve lesions which caused irreversible damage the microglial cells encircled the degenerating neurons and ultimately phagocytosed them. After crush lesions there were great species variations in the glial reaction: the neurons of rats and rabbits showed considerable and reversible separation of the synaptic terminals by glial cells, while mice showed no significant synaptic separation. It is suggested that the main function of the microglial cells may be to serve as potential phagocytes for disintegrating neurons, and that the reversible glial changes around regenerating neurons possibly represent abortive glial reactions.

Age Factors

Type 2 diabetes genetics in 125,000 admixed adults from Mexico City.

Type 2 diabetes (T2D) is a highly heritable, polygenic disease with over 600 loci identified through genome-wide association studies (GWAS). However, despite possessing unique genetic variation shaped by demographic history and admixture, Latin American populations remain markedly underrepresented in global genomic research. To address this gap, we conducted genome- and exome-wide analyses of 19,431 T2D cases and 105,611 controls from the Mexico City Prospective Study (MCPS). We identified 86 independent GWAS associations, including 21 novel signals, 15 of which replicated in external cohorts. Risk alleles at novel loci were enriched in individuals with Indigenous American ancestry. Exome analyses revealed rare and ultra-rare missense variants with substantial risk effects at HNF1A and GCK, as well as a protein-damaging variant in SLC30A8 that reduced T2D risk by 45% in carriers. Integrative analyses indicate that T2D genetic architecture in Mexico is predominantly driven by common regulatory variation acting in the endocrine pancreas. Polygenic risk scores strongly stratified T2D risk and transferred to Indigenous Mexican populations. These findings demonstrate the power of large-scale genetic discovery in diverse populations to refine disease architecture and identify loci with potential therapeutic relevance.

Journal Article

SLC1A5 and NUMA1 are potential regulators and therapeutic targets of ferroptosis in diffuse large B-cell lymphoma.

BACKGROUND: Ferroptosis, a form of regulated cell death driven by iron-dependent lipid peroxidation, has emerged as a potential therapeutic target in various cancers, including diffuse large B-cell lymphoma (DLBCL). This study aimed to identify and characterize ferroptosis-related panel genes with prognostic value in DLBCL. METHODS: Transcriptomic data from Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) were analyzed to identify differentially expressed genes (DEGs) in DLBCL samples. Gene set variation analysis (GSVA) and network topology analysis were performed to identify key ferroptosis-related genes. Lasso regression was utilized to construct a prognostic model based on the identified panel genes. In vitro experiments, including gene silencing, overexpression, and ferroptosis induction, were conducted to evaluate the functional roles of the identified genes, NUMA1 and SLC1A5, in DLBCL cells. RESULTS: A panel of ferroptosis-related genes with prognostic value, including NUMA1 and SLC1A5, was identified in DLBCL samples. Silencing SLC1A5 or overexpressing NUMA1 in DLBCL cells enhanced sensitivity to ferroptosis inducers, increased intracellular labile iron and lipid peroxidation levels, promoted mitochondrial damage, and modulated the expression of key ferroptosis markers. Furthermore, SLC1A5 silencing or NUMA1 overexpression augmented radiation-induced ferroptosis in DLBCL cells. CONCLUSION: NUMA1 and SLC1A5 are potential ferroptosis regulators and therapeutic targets in DLBCL. Silencing the ferroptosis-suppressive transporter SLC1A5 or restoring NUMA1 expression promotes lipid peroxidation and ferroptotic cell death, thereby sensitizing DLBCL cells to ferroptosis and enhancing radiosensitivity-providing a rationale for novel ferroptosis-based therapeutic strategies.

Humans

Osteoarthritis phenotypes: advancing precision medicine through clinical, structural, and molecular stratification.

PURPOSE: Osteoarthritis (OA) is now understood as a heterogeneous syndrome driven by diverse biological, biomechanical, metabolic, genetic, and molecular mechanisms. This variability explains differences in disease progression and treatment response, challenging the traditional "one-size-fits-all" approach. This review highlights OA phenotyping as a key step toward precision medicine, focusing on clinical, structural, and molecular classifications that inform individualized care. METHODS: A narrative review was conducted using a non-systematic search of major databases and Osteoarthritis Research Society International sources (2010-2026). Evidence was thematically synthesized across clinical, imaging, and molecular domains to characterize OA phenotypes and their potential relevance to precision medicine. RESULTS: Multiple OA phenotypes were identified: inflammatory, metabolic, biomechanical, cartilage-subchondral, pain-sensitization, and aging/senescence. These exhibit distinct clinical features, risk factors, and therapeutic responses. Imaging-based phenotypes (e.g., inflammatory, meniscus-cartilage, subchondral bone, atrophic, hypertrophic) and molecular endotypes (low turnover, structural damage, systemic inflammation) further refine stratification. Pain-structure discordance is notable in sensitization phenotypes and may predict poorer surgical outcomes. Joint-specific variations and emerging genomic and epigenetic insights underscore disease complexity. Advances in imaging, biomarkers, and machine learning may enable earlier detection and patient clustering, though clinical application remains limited. CONCLUSION: Phenotype- and endotype-based classification represents a critical advancement toward precision OA management. Tailored interventions based on stratification hold promise for improving outcomes; however, clinical translation remains limited by overlapping phenotypes, lack of validated biomarkers, and inconsistent results from phenotype-driven trials. Wider clinical adoption requires standardized definitions, validation across joints, and integration of multimodal diagnostic tools into routine practice.

Humans

Studies on acute and late stages of experimental central retinal artery occlusion in the Cynomolgus monkey. II. Influence on the cyclic changes in the amplitude of the c-wave of the ERG and in the standing potential of the eye.

The slow (2-3/h) oscillations of the c-wave amplitude and of the standing potential of the eye (SP) were studied in the Cynomolgus monkey during the acute and late stages after experimentally induced (laser photocoagulation) occlusion of the central retinal artery (OCRA). Whereas the healthy control eyes showed large cyclic variations in both the c-wave amplitude and the SP, no oscillations in the c-wave amplitude were observed in the OCRA eyes at any stage, and the SP oscillations were barely detectable in these eyes. OCRA causes morphological damage to the inner retina but not to the pigment epithelium-photoreceptor complex, which generates the positive component of the c-wave, and where the SP is also believed to originate. The findings of the present study strongly indicate that the SP and c-wave oscillations are related, and that they are either dependent on an intact inner retina or that the pigment epithelium-photoreceptor complex is functionally affected by OCRA. If not taken into consideration, the marked difference in oscillations between the OCRA eye and the healthy eye may be a major source of error when comparing c-waves from the two eyes, and it seems that some of the conflicting results reported by others can be explained in this way.

Acute Disease

Extensive longevity and DNA virus-driven adaptation in nearctic Myotis bats.

The genus Myotis is one of the largest clades of bats, and exhibits some of the most extreme variation in lifespans among mammals alongside unique adaptations to viral tolerance and immune defense. To study the evolution of longevity-associated traits and infectious disease, we generated cell lines and near-complete genome assemblies for 8 closely related species of Myotis. Using genome-wide screens of positive selection, analyses of structural variation, and functional experiments in primary cells, we identify new patterns of adaptation contributing to longevity, cancer resistance, and viral interactions in bats. We show that the recurrent evolution of longevity seen in Myotis leads to some of the highest predicted increases in cancer risk across mammals and demonstrate a unique DNA damage response in primary cells of the long-lived M. lucifugus. We also find evidence of abundant adaptation in response to DNA viruses - but not RNA viruses - in Myotis and other bats in sharp contrast with other mammals, potentially contributing to the role of bats as reservoirs of zoonoses. Together, our results demonstrate how genomics and primary cells derived from diverse taxa uncover the molecular bases of extreme adaptations in non-model organisms.

Aging

Postlaminectomy arteriovenous fistula.

Forty-nine cases of postlaminectomy arteriovenous fistula have been reported. Perforation of the anterior spinal ligament by the pituitary rongeur during discectomy with resultant simultaneous damage to artery and vein is causative. Whether aorta, cava, and/or iliac vessels are involved depends upon the level of laminectomy, the angle of the instrument, and anatomic variations in aortocaval bifurcation. Nine patients presented early after operation and were diagnosed promptly. The 40 patients who presented from months to years following laminectomy to physicians unfamiliar with this entity often had a distressing delay in diagnosis. High output congestive heart failure, particularly in a young person, and the characteristic abdominal and back bruit, should arouse suspicion. Arteriography confirms the diagnosis and allows planning for the operative repair. Though potentially disastrous, the operative correction of a major arteriovenous fistula may be done safely if standard principles of vascular surgery are followed. When combined with technical hints regarding clamp placement, transvascular repair, balloon catheter use, vein preservation, and the multiple inventive uses of the Dacron vascular prosthesis, a successful outcome should be expected.

Adult