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Biomedical subjects

C Yang

Publications and source records attributed to C Yang.

At least 73 records · Page 4Linked to original sources
PubMed2001

Developmental changes in the expression of GABA(A) receptor subunits (alpha(1), alpha(2), alpha(3)) in the cat visual cortex and the effects of dark rearing.

The present study used Western blots and Northern slot blots to determine changes in the level of expression of GABA(A) receptor subunits alpha(1), alpha(2), and alpha(3), in relation to the "critical period" in cat visual cortex. Levels of the GABA(A) alpha(1) subunit were lowest at 1 week, increased four-fold to a maximum at 10 weeks, and declined slightly (35%) into adulthood. Levels of the GABA(A) alpha(2) and alpha(3) subunits were highest at 1 week of age, decreased two-fold by 10 weeks of age and were constant thereafter. Comparison between visual cortex from normal and dark-reared cats at 5 weeks and 20 weeks showed that alpha(1) and alpha(3) subunit expression was elevated in dark-reared animals by approximately 50% at both ages. alpha(2) expression was not affected. These results implicate the importance of a shift from putative immature to mature GABA(A) receptor subunits during the critical period of visual cortex and in conjunction with parallel analysis of NMDA receptor subunit maturation, further support the notion that a changing excitatory/inhibitory balance is critical for neuronal plasticity.

Animals↗
PubMed2001

Predominance of HIV type 1 subtype G among commercial sex workers from Kinshasa, Democratic Republic of Congo.

We have investigated the genetic diversity and potential mosaic genomes of HIV-1 during the early part of the HIV-1 epidemic among commercial sex workers (CSWs) in Kinshasa, Democratic Republic of Congo (formerly Zaire). Serologic analysis revealed that 27 (28.7%) of the 94 specimens were seropositive by both peptide and whole-virus lysate EIAs and that 24 were positive by molecular screening assays, using generic primers that can detect all known groups of HIV-1. Phylogenetic analyses of the gag(p24), C2V3, and gp41 regions of these 24 specimens showed that all were group M; none of them had any evidence of group O, N, or SIVcpz-like sequences. On the basis of env sequence analysis, the 24 group M specimens were classified as subtypes G (37.5%), A (21%), F1 (12.5%), CRF01_AE (8%), D (4%), and H (4%); 3 (12.5%) were unclassifiable (U). Similar analysis of the gag(p24) region revealed that the majority of infections were subtype A; however, one-third of the specimens were subtype G. Parallel analysis of gag(p24) and env regions revealed discordant subtypes in many specimens that may reflect possible dual and/or recombinant viruses. These data suggest a predominance of subtype G (both pure G and recombinant CRF02_AG) during the early part of the epidemic in Kinshasa. Infections with group N or SIVcpz-like viruses were not present among these CSWs in Kinshasa.

Democratic Republic of the Congo↗
PubMed2001

Relation between pediatric experience and treatment recommendations for children and adolescents with kidney failure.

CONTEXT: Children and adolescent patients with renal failure are frequently cared for by adult subspecialists. While peritoneal dialysis is used in less than 17% of adults with kidney failure, it is the preferred dialysis treatment for children. National data show that 45% of children receiving dialysis are treated with peritoneal dialysis and pediatric nephrologists report its use in 65% of patients receiving dialysis. Whether differences in peritoneal dialysis use among children are due to the pediatric experience of the clinician has not been examined. OBJECTIVE: To assess whether the pediatric experience of nephrologists directly affects treatment recommendations for children with kidney failure. DESIGN: Cross-sectional survey using 10 case vignettes per survey based on random combinations of 8 patient characteristics (age, sex, race, distance from facility, cause of renal failure, family structure, education, and compliance). SETTING AND PARTICIPANTS: National random sample of office-, hospital-, and academic medical center-based adult and pediatric nephrologists, stratified by geographic region and conducted June to November 1999. Of 519 eligible physicians, 316 (61%) responded, including 191 adult and 125 pediatric nephrologists. MAIN OUTCOME MEASURE: Treatment recommendations for peritoneal dialysis vs hemodialysis, compared based on nephrologists' pediatric experience. RESULTS: After controlling for patient characteristics, pediatric nephrologists were 60% more likely than adult nephrologists to recommend peritoneal dialysis for identical patients (odds ratio, 1.61; 95% confidence interval, 1.35-1.92). This was true regardless of dialysis training, years in practice, practice setting, geography, or patient characteristics. CONCLUSIONS: Our data indicate that pediatric specialization of clinicians influences treatment recommendations for children and adolescents with end-stage renal disease. Referring children to adult subspecialists may lead to differences in treatment choices and processes of care.

Adolescent↗
PubMed2001

Cloning and characterization of full-length human ribosomal protein L15 cDNA which was overexpressed in esophageal cancer.

The aim of this investigation was trying to identify the genes differentially expressed in esophageal cancer. By combining suppression subtractive hybridization (SSH) with reverse Northern high density blots, a gene named EC45 was obtained, which dramatically overexpressed in 70% esophageal cancer (18/26). EC45 was mapped to 3p12-3p11.2 by radiation hybrid mapping (RH mapping). The putative full length EC45 cDNA (1987 bp) was identified by cDNA libraries screening of esophageal cancer. EC45 encoded 204 amino acids, and it shared a 100% similarity with ribosomal protein L15 (635 bp, mRNA) in ORF, but no similarity in 5' UTR or 3' UTR. Northern blot panel of multiple adult human normal tissues showed EC45 distributed in almost normal tissues tested. All these data suggested that EC45, encoding ribosomal protein L15 and overexpressing in esophageal cancer might play a possible role in carcinogenesis of esophagus.

Amino Acid Sequence↗
PubMed2001

Early life stress and inherited variation in monkey hippocampal volumes.

BACKGROUND: Opportunities for research on the causes and consequences of stress-related hippocampal atrophy are limited in human psychiatric disorders. Therefore, this longitudinal study investigated early life stress and inherited variation in monkey hippocampal volumes. METHODS: Paternal half-siblings raised apart from one another by different mothers in the absence of fathers were randomized to 1 of 3 postnatal conditions that disrupted diverse aspects of early maternal care (n = 13 monkeys per condition). These conditions were previously shown to produce differences in social behavior, emotional reactivity, and neuroendocrine stress physiology. Hippocampal volumes were subsequently determined in adulthood by high-resolution magnetic resonance imaging. RESULTS: Adult hippocampal volumes did not differ with respect to the stressful postnatal conditions. Based on paternal half-sibling effects, the estimated proportion of genetic variance, ie, heritability, was 54% for hippocampal size. Paternal half-siblings with small adult hippocampal volumes responded to the removal of all mothers after weaning with initially larger relative increases in cortisol levels. Plasma cortisol levels 3 and 7 days later, and measures of cortisol-negative feedback in adulthood were not, however, correlated with hippocampal size. CONCLUSIONS: In humans with mood and anxiety disorders, small hippocampal volumes have been taken as evidence that excessive stress levels of cortisol induce hippocampal volume loss. Results from this study of monkeys suggest that small hippocampi also reflect an inherited characteristic of the brain. Genetically informed clinical studies should assess whether inherited variation in hippocampal morphology contributes to excessive stress levels of cortisol through diminished neuroendocrine regulation.

Animals↗
PubMed2001

Nasal absorption of tyrosine-linked model compounds.

The objective of this research is to explore the amino acid carrier-mediated pathway for nasal uptake of tyrosine-linked model compounds. These molecules were synthesized by ether linkage between the aromatic hydroxyl group of L-tyrosine and the terminal hydroxyl group of model compounds. An enzymatically stable linkage was utilized to link the model compounds to L-tyrosine so that the loss of the compound from perfusate signals absorption only without complication from metabolism. The rat in situ nasal perfusion technique was utilized in this investigation. The analyte concentrations remaining in the nasal perfusates were quantitated by a reversed-phase high-performance liquid chromatography. All L-tyrosine-linked model compounds were absorbed across the nasal mucosa (15-50% absorbed), and the extent of nasal absorption was concentration dependent. The apparent first-order rate constants of intact conjugate loss from the nasal perfusate were calculated. The Michaelis-Menten kinetic model was applied to fit the perfusion data. The Michaelis constant, K(m), and maximum or saturation rate of compound loss from the perfusate, V(max), were calculated from Lineweaver-Burk plots [e.g., K(m) and V(max) are 0.57 mM and 0.00515 micromol/min, respectively, for one of the model compounds, O-(4-carboxyphenylmethyl)-L-tyrosine]. The absorption characteristics of all these model compounds exhibited saturable kinetics, whereas their parent compounds, possessing different octanol-water partition coefficients, showed either no absorption or transport by passive diffusion. Nasal uptake of L-tyrosine conjugates was inhibited by L-tyrosine. The results from this study indicate that nasal absorption of L-tyrosine-linked compounds is probably through an amino acid transport system. Exploitation of the amino acid transport pathway may be utilized to improve nasal absorption of poorly permeable drugs.

Administration, Intranasal↗
PubMed2001

Chemical stability, enzymatic hydrolysis, and nasal uptake of amino acid ester prodrugs of acyclovir.

The objective of this work was to improve nasal absorption of relatively impermeable small drug molecules via an amino acid prodrug approach. Acyclovir was selected as a model drug. L-Aspartate beta-ester, L-lysyl, and L-phenylalanyl esters of acyclovir were synthesized to investigate their effectiveness in enhancing nasal absorption of acyclovir. A stability study was conducted in phosphate buffer under various pH conditions at 25 and 37 degrees C. Enzymatic hydrolysis in rat nasal washings and plasma was conducted at 37 degrees C. A rat in situ nasal perfusion technique was utilized in this investigation to examine the rate and extent of nasal absorption of amino acid prodrugs. The remaining analyte concentrations in the nasal perfusate were quantitated by reversed-phase high-performance liquid chromatography. The results revealed that the L-lysyl and L-phenylalanyl esters were less stable than L-aspartate beta-ester. The stability of all three esters decreased with increasing pH and temperature. L-phenylalanyl ester is highly susceptible to plasma esterases, with an in vitro half-life 1.33 min. The rat in situ nasal perfusion study revealed that the extent of nasal absorption of acyclovir, L-lysyl and L-phenylalanyl esters was not significant (p < 1%). L-Aspartate beta-ester was absorbed to the extent of approximately 8% over 90 min of perfusion at an initial drug concentration of 100 microM. Nasal absorption of L-aspartate beta-ester of acyclovir was inhibited by L-asparagine but not by a dipeptide glycylsarcosine (Gly-Sar). The enhancement of acyclovir nasal absorption from the L-aspartate beta-ester prodrug suggests that nasal uptake of this prodrug probably involves an active transport system.

Acyclovir↗
PubMed2001

Both CD45R(low) and CD45R(high) "revertant" CD4 memory T cells provide help for memory B cells.

During a primary response to thymus dependent antigens, B cells undergo a number of qualitative changes to become memory B cells - processes that require co-stimulatory signals and cytokine help from CD4 T cells. The question of whether distinct, antigen-experienced memory CD4 T cells are subsequently needed to program memory B cells into antibody synthesis has not been clearly resolved. Using an adoptive transfer model in which memory but not naive B cells were stimulated, we evaluated CD4 T cell help using lymphocytes obtained from primed or unprimed thymectomized donors and expressing a naive (CD45R(high)) or a memory (CD45R(low)) phenotype. Memory B cells, most of which were committed to the IgG1 (Th2) subclass, could be stimulated to produce antibody using help transferred by the CD45R(high) naive subset of unprimed donors (slow onset of response), the CD45R(low) subset of 7 day primed donors (large, rapid antibody response) or by both the CD45R(low) and the CD45R(high) "revertant" subsets of 6 month primed donors. We found that antigen primed CD45R(low) CD4 T cells reverted (defaulted) with time to a CD45R(high) resting state, a change that was prevented by persisting antigen. The evidence suggests that CD4 memory T cells are partitioned into two different functional states (CD45R(high) and CD45R(low)) and that these determine the characteristics of the memory B cell response in terms of speed, size and longevity.

Animals↗
PubMed2001

Hypersensitivity pneumonitis in a metal-working environment.

BACKGROUND: An outbreak of lung disease among workers in a metal-working plant included 16 biopsy-confirmed cases of hypersensitivity pneumonitis and additional patients with asthma, bronchiolitis and emphysema, usual interstitial pneumonitis, and sarcoidosis. Study design Clinical examination of patients; cross-sectional questionnaire survey of the outbreak plant and two control plant areas, one with and one without MWF exposures, in a separate facility; industrial hygiene survey with laboratory characterization of microbial flora; and immunological investigation. METHODS: Patients with suspected hypersensitivity pneumonitis underwent a clinical examination including detailed lung function, imaging, and tissue studies. A plant walk-through identified metal-working processes, microbial aerosols, and work practices. Microbial characteristics of the three microbial aerosol-producing processes were characterized. Antibodies to those agents were determined in patient sera. A questionnaire survey was conducted in the case plant and in two areas of a control plant, one with and one without metal-working fluids exposure. RESULTS: Thirty-nine (79.6%) patients described symptoms consistent with work-related lung disease, eight received other diagnoses, and two did not complete their examinations. Sixteen patients had hypersensitivity pneumonitis confirmed on biopsy. Mean decrements in lung forced expiratory volume in 1 s and force vital capacity from before to after work were similar in the 16 biopsy-confirmed cases of hypersensitivity pneumonitis ( - 6.3%; - 7.2%) and the 19 symptomatic patients without biopsies ( - 11.2%, - 10.1%). Symptoms were more common in the case plant than in a non-MWF control plant area. Three sources of water-based aerosols were identified that grew similar microbial flora. Although machining increased airborne bacterial levels, the increase was not related to the concentration of viable bacteria in the sumps. Antibody testing did not identify a specific single organisms. Endotoxin levels were similar in case and MWF control plant. CONCLUSIONS: Lung disease in environments with water-based aerosols may be more common than usually recognized. Patients with HP often present with only subtle abnormalities and may be missed if multiple clinical abnormalities are required to document disease.

Adult↗
PubMed2001

Stereoselective reduction with NADH model BNAH through chiral induction in cyclodextrins.

Stereoselective reductive debromination-cyclopropanation of 2-bromo-1-phenylethylidene-malononitrile and 2-bromo-1-beta-naphthylethylidenemalononitrile by coenzyme NADH model BNAH through chiral induction in cyclodextrins is reported. The matching between substrates and cyclodextrins, the substituent effect, and the effect of cyclodextrin concentration on the optical yields have been investigated.

Cyclodextrins↗
PubMed2001

Human vascular endothelial growth factor cDNA cloning and expression in osteoblasts.

Human vascular endothelial growth factor (VEGF) cDNA was amplified by nested polymerase chain reaction method from the HL60 cells. Then a pCD-hVEGF165 recombinant plasmid was constructed. Rabbit osteoblasts were transfected with pCD-hVEGF165 plasmid by lipofectin mediated gene transfer. The transient expressive results were detected by immunohistochemical method. It was observed that the expression of human VEGF gene was detected 72 h after transfecting distinctly.

Animals↗
PubMed2001

Bladder neck circulation by Doppler ultrasonography in postmenopausal women with urinary stress incontinence.

OBJECTIVE: To investigate the effect of hormone replacement therapy (HRT) on bladder neck circulation in postmenopausal women with genuine urinary stress incontinence (USI). METHODS: A total of 227 women were enrolled in the study, including 114 postmenopausal women with USI (n = 57) or without USI (n = 57) and 113 premenopausal women with USI (n = 55) or without USI (n = 58). The bladder neck circulation was measured in 31 postmenopausal women with USI and in 12 without USI, all of whom received 0.625 mg conjugated equine estrogen plus 5 mg medroxyprogesterone acetate daily for 6 months. Bladder neck blood flow data detected by perineal color Doppler ultrasonography and weekly urinary diary data were collected before HRT and then 3 and 6 months after beginning HRT. Two-way analysis of variance and repeated measures of variance were used for statistical analysis. Power analysis was done by a two-sided test with a significance level at.05. RESULTS: The pulsatility index (PI) was significantly higher in the postmenopausal than in the premenopausal women. The postmenopausal women with USI had the highest PI level. The presence of USI did not change the PI values in the premenopausal women. After 3 months of HRT, the PI levels decreased significantly (P <.001) in the postmenopausal women with USI. The subjective improvement of USI appeared after 3 months of HRT. CONCLUSION: The blood flow around the bladder neck in women can be measured by perineal color Doppler ultrasonography. Hormone replacement therapy increases the blood flow around the bladder neck in postmenopausal women with USI. The clinical improvement of USI can be seen with HRT after 3 months.

Female↗
PubMed2001

Mechanisms of bradykinin-mediated Ca(2+) signalling in canine cultured corneal epithelial cells.

Experiments were designed to differentiate the mechanisms of bradykinin receptors mediating the changes in intracellular Ca(2+) concentration ([Ca(2+)](i)) in canine cultured corneal epithelial cells (CECs). Bradykinin and Lys-bradykinin caused an initial transient peak of [Ca(2+)](i) in a concentration-dependent manner, with half-maximal stimulation (pEC(50)) obtained at 6.9 and 7.1, respectively. Pretreatment of CECs with pertussis toxin (PTX) or cholera toxin (CTX) for 24 h did not affect the bradykinin-induced [Ca(2+)](i) changes. Application of Ca(2+) channel blockers, diltiazem and Ni(2+), inhibited the bradykinin-induced Ca(2+) mobilization, indicating that Ca(2+) influx was required for the bradykinin-induced responses. Addition of thapsigargin (TG), which is known to deplete intracellular Ca(2+) stores, transiently increased [Ca(2+)](i) in Ca(2+)-free buffer, and subsequently induced Ca(2+) influx when Ca(2+) was readded to this buffer. Pretreatment of CECs with TG completely abolished bradykinin-induced initial transient [Ca(2+)](i), but had slight effect on bradykinin-induced Ca(2+) influx. Pretreatment of CECs with 1-[beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole (SKF96365) and 1-(6-((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122) inhibited the bradykinin-induced Ca(2+) release and Ca(2+) influx, consistent with the inhibition of receptor-gated Ca(2+) channels and phospholipase C (PLC) in CECs, respectively. These results demonstrate that bradykinin directly stimulates B(2) receptors and subsequently Ca(2+) mobilization via a PTX-insensitive G protein in canine CECs. These results suggest that bradykinin-induced Ca(2+) influx into the cells is not due to depletion of these Ca(2+) stores, as prior depletion of these pools by TG has no effect on the bradykinin-induced Ca(2+) influx that is dependent on extracellular Ca(2+) in CECs.

Animals↗
PubMed2001

Gelatinase A (MMP-2) activation by skin fibroblasts: dependence on MT1-MMP expression and fibrillar collagen form.

The respective requirements of collagen and MT1-MMP in the activation of MMP-2 by primary fibroblast cultures were explored further. Three-dimensional gels enriched in human collagen types I and III or composed of recombinant human type II or III collagen, caused increased MT1-MMP production (mRNA and protein) and induced MMP-2 activation. Only marginal induction was seen with dried monomeric collagen confirming the need for collagen fibrillar organisation for activation. To our surprise, relatively low amounts (as low as 25 microg/ml) of acid soluble type I collagen added to fibroblast cultures also induced potent MMP-2 activation. However, the requirement for collagen fibril formation by the added collagen was indicated by the inhibition seen when the collagen was pre-incubated with a fibril-blocking peptide, and the reduced activation seen with alkali-treated collagen preparations known to have impaired fibrilisation. Pre-treatment of the collagen with sodium periodate also abrogated MMP-2 activation induction. Further evidence of the requirement for collagen fibril formation was provided by the lack of activation when type IV collagen, which does not form collagen fibrils, was added in the cultures. Fibroblasts derived from MT1-MMP-deficient mice were unable to activate MMP-2 in response to either three-dimensional collagen gel or added collagen solutions, compared to their littermate controls. Collectively, these data indicate that the fibrillar structure of collagen and MT1-MMP are essential for the MMP-2 activational response in fibroblasts.

Animals↗
PubMed2001

Modulation of aromatase expression in human breast tissue.

Aromatase plays an important role in breast cancer development through its role in the synthesis of estrogen. Aromatase expression in breast tissue can be regulated by several mechanisms. The major promoter usage for aromatase expression in breast tumors (i.e. cAMP-stimulated promoters I.3 and II) is different from that in normal breast tissue (i.e. glucocorticoid-stimulated promoter I.4). Recent characterization of transcription factors that interact with the two important regulatory elements near promoters I.3 and II, i.e. S1 and CREaro, helps us better understand the mechanism of the switch of promoter usage between normal breast tissue and cancer tissue. It is thought that in normal breast tissue, the function of promoters I.3 and II is suppressed through the binding of EAR-2, COUP-TFI, and EARgamma to S1, and through the binding of Snail/Slug proteins to their binding site that quenchs the CREaro activity. In cancer tissue, the expression levels of EAR-2, COUP-TFI, EARgamma, Snail, and Slug decrease, and aromatase expression is then up regulated through the binding of ERRalpha-1 to S1 and the binding of CREB or related factors to CREaro. Results from this and other laboratories reveal that aromatase activity in aromatase expressing cells can also be modified by treatment with aromatase inhibitors and the antiestrogen ICI 182, 780. While aromatase inhibitors are used to treat breast cancer, the treatment has been found to increase the level of aromatase in the breast tissue of some patients. The enhancement of aromatase activity by aromatase inhibitors is thought to be due to a decrease of aromatase protein degradation by enzyme-inhibitor complex formation, up-regulation of the aromatase gene transcription through a cAMP-mediated mechanism, and an induction of aromatase expression by gonadtropins that are released from the pituitary in response to a reduction of estrogen levels in circulation in premenopausal women. Antiestrogen ICI 182, 780 has been found to suppress aromatase expression, but the mechanism has not yet been determined. In addition, aromatase activity and expression can be affected by environmental chemicals. A detailed structure-function study has revealed that flavones, but not isoflavones, are inhibitors of aromatase. It was found that flavones bind to the active site of aromatase in an orientation in which their rings-A and -C mimic rings-D and -C of the androgen substrate. The modulation of aromatase expression by endocrine disrupting chemicals is exemplified by two organochlorine pesticides (i.e. toxaphene and chlordane) that have been found to be antagonists of ERRalpha-1 orphan receptor. These compounds reduce ERRalpha-1 activity, resulting in a suppression of aromatase expression.

Aromatase↗
PubMed2001

Sonochemical degradation for toxic halogenated organic compounds.

This paper describes the degradation of p-chlorophenol using three different ultrasonic devices. The dissipated power in the reaction matrix was measured based on calorimetric method. The study showed that hydrogen peroxide can improve the sonochemical reaction and gases dissolved in reaction matrix can affect the process to a small extent. The reaction mechanism and kinetics of degradation were also investigated.

Calorimetry↗
PubMed2001

Volatiles from Ficus hispida and their attractiveness to fig wasps.

Volatile compositions of receptive (ready to be pollinated), postpollinated, and postparasitized figs. and leaves of Ficus hispida were analyzed. Differences among them were examined, and the specificity of fig wasp attractiveness was investigated. Linalool was the major constituent of steam-distilled oil of either male or female receptive figs, while dibutyl phthalate was the major compound of the oils of postparasitized and postpollinated figs. In petroleum ether extracts, palmitic oil, and 9,12-octadecadienoic acid were the main constituents of male and female receptive figs, while hexadecanoic acid ethyl ester was the major compound of postparasitized and postpollinated figs. In dichloromethane extracts, linalool was the major constituent of male and female receptive figs, 1-hydroxylinalool was the major component of male postparasitized figs, and 1-hydroxylinalool and benzyl alcohol were the major constituents of female postpollinated figs. Bioassays with sticky traps showed that Ceratosolen solmsimarchal was attracted to dichloromethane extracts of male and female receptive figs and to petroleum ether extracts of female receptive figs, but was not attracted to dichloromethane and petroleum ether extracts of male postparasitized and female postpollinated figs. Figs were attractive to pollinating wasps only at the receptive stage. The volatile constituents of receptive figs were different from those of postpollinated or postparasitized figs. From a receptive to a postpollinated state, figs changed in their volatile composition. Some compounds disappeared or decreased in amount. These include linalool, linalool oxide, alpha-terpeneol, and 2,6-dimethyl-1,7-octadiene-3,6-diol, which may act as the attractants of the wasps. Others increased in amount, or several additional chemicals appeared. These include dibutyl phthalate, 1-hydroxylinalool, and benzyl alcohol, which may be repellents of the wasps. That dichloromethane extracts of male and female receptive figs showed similar activities in attracting fig wasps indicates that receptive figs of both sexes are similarly attractive to fig wasps, which is further supported by their similar volatile composition. Leaf extract was not attractive to the wasps.

Animals↗