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U Gehring

Publications and source records attributed to U Gehring.

At least 19 recordsLinked to original sources
PubMed2002

Predictors of high endotoxin concentrations in the settled dust of German homes.

Endotoxin concentrations in settled house dust were measured within the collaborative study named INGA (Indoor Exposure and Genetics in Asthma) together with other asthma-related indoor factors in 405 randomly selected homes in two German cities. Endotoxin was assayed using the quantitative kinetic chromogenic Limulus Amebocyte Lysate (LAL) method. The mean concentration of 2,274 ng endotoxin/g dust in living rooms is about two orders of magnitude below mean occupational exposure (e.g. agriculture). Endotoxin concentrations are higher in old buildings (means ratio = (MR) 1.52, 95% C.I.: 1.14; 2.04), and in the lower storey of the dwelling (MR = 1.30, 95% C.I.: 1.04; 1.62). Furthermore, higher endotoxin concentrations are associated with longer occupancy in the apartment (MR = 1.63, 95% C.I.: 1.21; 2.18), high utilisation of the apartment during the sampling period (MR = 2.52, 95% C.I.: 1.42; 4.47), infrequent vacuum cleaning of the carpets (MR = 1.67; 95% C.I.: 1.10; 2.54), an indifferent attitude to ventilation (MR = 1.37; 95% C.I.: 1.03; 1.82), keeping cats (MR = 1.91; 95% C.I.: 1.43; 2.55) and dogs (1.57; 95% C.I.: 1.08; 2.30), as well as the occurrence of mice (MR = 1.39; 95% C.I.: 1.00; 1.93). The season and indoor climate (operative temperature and relative humidity) did not effect endotoxin concentration. These results indicate that high endotoxin concentration in settled dust is an indicator of poor hygienic conditions in homes.

Adult↗
PubMed2002

Maternal compliance with nutritional recommendations in an allergy preventive programme.

AIMS: To assess maternal compliance with nutritional recommendations in an allergy preventive programme, and identify factors influencing compliance behaviour. METHODS: Randomised double-blind intervention study on the effect of infant formulas with reduced allergenicity in healthy, term newborns at risk of atopy. Maternal compliance with dietary recommendations concerning milk and solid food feeding was categorised. RESULTS: A total of 2252 newborns were randomised to one of four study formulas. The drop out rate during the first year of life was 13.5% (n = 304). The rates of high, medium, and low compliance to milk feeding during weeks 1-16 were 83.4%, 4.0%, and 7.5%; the corresponding rates to solid food feeding during weeks 1-24 were 60.0%, 12.1%, and 22.9%. In 5.1% of subjects no nutritional information was available. Low compliance was more frequent among non-German parents, parents with a low level of education, young mothers, smoking mothers, and those who weaned their infant before the age of 2 months. CONCLUSIONS: Evaluation of allergy preventive programmes should take into account non-compliance for assessing the preventive effectiveness on study outcome.

Adult↗
PubMed2002

Traffic-related air pollution and respiratory health during the first 2 yrs of life.

As part of an international collaborative study on the impact of Traffic-Related Air Pollution on Childhood Asthma (TRAPCA), the health effects associated with long-term exposure to particles with a 50% cut-off aerodynamic diameter of 2.5 microm (PM2.5), PM2.5 absorbance, and nitrogen dioxide (NO2) were analysed. The German part of the TRAPCA study used data from subpopulations of two ongoing birth cohort studies (German Infant Nutrition Intervention Programme (GINI) and Influences of Lifestyle Related Factors on the Human Immune System and Development of Allergies in Children (LISA)) based in the city of Munich. Geographic information systems (GIS)-based exposure modelling was used to estimate traffic-related air pollutants at the birth addresses of 1,756 infants. Logistic regression was used to analyse possible health effects and potential confounding factors were adjusted for. The ranges in estimated exposures to PM2.5, PM2.5 absorbance, and NO2 were 11.9-21.9 microg m(-3), 1.38-4.39 x 10(-5) m(-1), and 19.5-66.9 microg x m3, respectively. Significant associations between these pollutants and cough without infection (odds ratio (OR) (95% confidence interval (CI)): 1.34 (1.11-1.61), 1.32 (1.10-1.59), and 1.40 (1.12-1.75), respectively) and dry cough at night (OR (95% CI): 1.31 (1.07-1.60), 1.27 (1.04-1.55), and 1.36 (1.07-1.74), respectively) in the first year of life were found. In the second year of life, these effects were attenuated. There was some indication of an association between traffic-related air pollution and symptoms of cough. Due to the very young age of the infants, it was too early to draw definitive conclusions from this for the development of asthma.

Air Pollution↗
PubMed2002

Predictors of cat allergen (Fel d 1) in house dust of German homes with/without cats.

BACKGROUND: Exposure to cat allergen is a major risk factor for sensitization and the development of asthma in many parts of the world. The study was designed to examine the levels of cat allergen (Fel d 1) in homes of two German cities and to detennine predictors of Fel d 1 exposure. METHODS: We collected dust samples from 405 randomly selected homes in Hamburg (n = 201) and Erfurt (n = 204). In each apartment dust samples were taken from living room floor (LR), bedroom floor (BR), and mattress surface (MA) using vacuum sampling and analyzed by two-site monoclonal antibody ELISA. Environmental variables were assessed by questionnaire to obtain information on factors supposed to have an impact on the Fel d 1 levels in house dust. The effects of possible predictors of Fel d 1 were assessed by multiple linear regression models. RESULTS: Fel d 1 was present in 98% of the homes (395/405), ranging from 0.015 to 7.278 microg/g dust (GM 0.486 microg/g). The levels were similar in floors (GM for LR 0.575 microg/g and BR 0.469 microg/g) and in MA (GM 0.424 microg/g). Higher allergen concentrations (> 2 microg/g) were detected in 99% of the homes keeping cats, in 55% of homes that had had a cat during the last year, and in 28% of the homes without a cat. Fel d 1 levels were significantly higher in homes with a cat (628-682-fold, MA, floor) and in homes in which a cat had previously lived (11-12-fold, floor, MA) than in homes that never had a cat. Furthermore, after controlling for possible other confounders, Fel d 1 levels were significantly higher in summer and in homes with low frequency of cleaning and low ventilation rate. CONCLUSION: It could be confirmed that keeping a cat has the highest impact on Fel d 1 concentration. Besides, a continuous influx of Fel d 1 in homes without cat, as a result of direct or indirect cat contact outside the home, is likely. High frequencies of dusting and ventilation might reduce cat allergens in homes with and without cats.

Allergens↗
PubMed2001

Prospective cohort studies using hydrolysed formulas for allergy prevention in atopy-prone newborns: a systematic review.

UNLABELLED: The aim of this study is to give a systematic overview on publications having investigated the allergy preventive effect of extensively and/or partially hydrolysed infant formulas. Publications were searched by several strategies. Inclusion criteria were: prospective cohort study, randomisation, family history of atopy of the subjects, follow-up for at least the 1st year of life. Studies were described systematically as to methods, study participants, interventions and co-interventions, and study outcomes. The methodological quality was judged according to an instrument assessing the reduction of bias (score between 0 and 7 points). Data of studies having examined partial hydrolysates are presented as odds ratios. Of 16 studies found, 10 fulfilled the inclusion criteria. Study designs showed considerable differences as to methods, intervention and co-intervention procedures and definition of outcomes. Methodological quality varied between 0 and 6 points. The comparison of exclusively test- and control formula-fed groups showed a uniform tendency towards allergy protection in studies with partial hydrolysates. Study results on the effect of extensive hydrolysates were not comparable because of major differences in study designs. CONCLUSION: More well-designed studies with an adequate statistical power are needed to compare the allergy preventive effect of partially and extensively hydrolysed formulas with a standard infant formula.

Cohort Studies↗
PubMed2001

The impact of draught related to air velocity, air temperature and workload.

This experimental study was designed to test the hypotheses that the effects of draught increase with higher air velocity, with lower air temperature, and with lower workload. Thirty healthy young males were exposed to horizontal draught during 55 min while they operated an arm ergometer in a standing posture. Air velocity, air temperature, and workload were varied in 3 steps each, between 11 and 23 degrees C, 0.1 and 0.3 m/s, and 104 to 156 W/m2, respectively. The 27 combinations were distributed over subjects in a fractional factorial 3(3)-design. The participants were clothed for thermal neutrality. Workload was measured at the end of the sessions by respirometry. Draught-induced annoyance was determined every 5 min, separately for 10 body sites. Corresponding skin temperature was also recorded. The hypotheses were verified for the influence of air velocity and air temperature. Regarding workload, local heat production is probably decisive, meaning that draft-induced local annoyance is inversely related to workload in active but independent from workload in non-active body areas. To improve the situation for the workers concerned it is suggested to apply protective gloves that cover an as great area of the forearms as possible and to limit airflows to mean velocities of less than 0.2 m/s (with turbulence intensities of 50%).

Adult↗
PubMed2001

Molecular characterization of a phosphatidylcholine-hydrolyzing phospholipase C.

While searching for a phospholipase C (PLC) specific for phosphatidylcholine in mammalian tissues, we came across such an activity originating from a contamination of Pseudomonas fluorescens. This psychrophilic bacterium was found to contaminate placental extracts upon processing in the cold. The secreted phosphatidylcholine-hydrolyzing PLC was purified by a combination of chromatographic procedures. As substrates, the enzyme preferred dipalmitoyl-phosphatidylcholine and 1-palmitoyl-2-arachidonoyl-phosphatidylcholine over phosphatidylinositol. The active enzyme is a monomer of approximately 40 kDa. As for other bacterial PLCs, the enzyme requires Ca2+ and Zn2+ for activity; dithiothreitol affected the activity due to its chelation of Zn2+, but this inhibition could be compensated for by addition of ZnCl2. The compound D609, described to selectively inhibit phosphatidylcholine-specific PLCs, caused half-inhibition of the P. fluorescens enzyme at approximately 420 microM, while 50-fold lower concentrations similarly affected PLCs from Bacillus cereus and Clostridium perfringens. Partial peptide sequences obtained from the pure P. fluorescens enzyme after tryptic cleavage were used to clone a DNA fragment of 3.5 kb from a P. fluorescens gene library prepared from our laboratory isolate. It contains an ORF of 1155 nucleotides encoding the PLC. There is no significant sequence homology to other PLCs, suggesting that the P. fluorescens enzyme represents a distinct subclass of bacterial PLCs. The protein lacks cysteine residues and consequently contains no disulfide bonds. Interestingly, P. fluorescens reference strain DSMZ 50090 is devoid of the PLC activity described here as well as of the relevant coding sequence.

Amino Acid Sequence↗
PubMed2001

Pets and vermin are associated with high endotoxin levels in house dust.

BACKGROUND: Previous studies have shown that the risk for allergic sensitization is lower in children who grew up on farms and in young adults who were exposed to dogs in early childhood. A higher microbial exposure in general and in particular to endotoxin in early childhood might contribute to this lower risk of atopy. OBJECTIVE: We examined whether the presence of pets or vermin in the home is associated with higher endotoxin concentrations in settled house dust. METHODS: House dust was sampled in a standardized manner on the living room floors of 454 homes of German children aged 5-10 years (participation rate 61%). Endotoxin was assessed with a quantitative kinetic chromogenic Limulus Amebocyte Lysate (LAL) method. Associations between endotoxin levels, pets and vermin are presented as ratios of the crude and confounder adjusted geometric means (means ratios) in the category of study vs. a reference category using multiple linear regression models. RESULTS: Endotoxin concentrations in living room floor dust sampled in homes without pets and vermin were lower (1246 ng per square meter, 1519 ng endotoxin/g dust, n = 157) than those sampled in homes with pets or vermin (2267 ng per square meter, 2200 ng endotoxin/g dust, n = 296). After adjustment for city of residence, season of dust sampling, age of the building and story of the dwelling, means ratios for endotoxin expressed per gram of dust were statistically significantly increased for dog (1.64, 95% CI 1.09-2.46), for cat (1.50, 95% CI 1.03-2.18) and for cockroach (3.01, 95% CI 1.37-6.60), whereas no major statistically significant associations were found for other pets, ants and mice. CONCLUSION: Keeping a dog or a cat in the home is consistent with higher exposure to endotoxin and might therefore contribute to the lower risk of atopy in later life.

Air Pollution, Indoor↗
PubMed2001

Exposure to endotoxin decreases the risk of atopic eczema in infancy: a cohort study.

BACKGROUND: Previous studies have shown a protective effect of early exposure to cats and dogs on the development of atopic eczema, asthma, allergic rhinitis, and atopic sensitization in later life. In particular, a higher microbial exposure to endotoxin in early childhood might contribute to this effect. OBJECTIVE: We examined the associations between bacterial endotoxin in house dust and atopic eczema, infections, and wheezing during the first year of life in an ongoing birth cohort study (LISA). METHODS: Data of 1884 term and normal-weight neonates with complete information on exposure to biocontaminants and confounding variables were analyzed. House dust from the mothers' and the children's mattresses was sampled 3 months after birth. Endotoxin content was quantified by using a chromogenic kinetic limulus amoebocyte lysate test. RESULTS: During the first 6 months of life, the risk of atopic eczema was significantly decreased by endotoxin exposure in dust from mothers' mattresses in the fifth quintile (odds ratio [OR], 0.50; 95% CI, 0.28-0.88), whereas the risk was increased for respiratory infections (OR, 1.69; 95% CI, 1.25-2.28) and cough with respiratory infection, bronchitis, or both (OR, 1.73; 95% CI, 1.28-2.33). The risk of wheezing was also significantly increased during the first 6 months of life (OR, 2.37; 95% CI, 1.40-4.03). For the entire first year of life, these associations attenuated, except for the risk of wheezing, which remained significant (OR, 1.60; 95% CI, 1.10-2.30). CONCLUSION: Our findings support the hygiene hypothesis that exposure to high concentrations of endotoxin very early in life might protect against the development of atopic eczema within the first 6 months of life, along with an increased prevalence of nonspecific respiratory diseases.

Air Pollution, Indoor↗
PubMed2001

Respiratory symptoms in relation to indoor exposure to mite and cat allergens and endotoxins. Indoor Factors and Genetics in Asthma (INGA) Study Group.

The authors investigated the relationship between respiratory symptoms in adults and exposure to mite and cat allergens, the role of endotoxins in house dust, the effects of mixtures of several allergens, and interactions between allergen exposure and allergic sensitization. Within a nested case-control study, 405 subjects aged 25-50 yrs from two German cities answered a standardized questionnaire. Allergen-specific immunoglobulin-E was measured. Dust samples were taken from the subjects' homes to determine exposure to mite (Dermatophagoides pteronyssinus antigen 1 Der p 1) and (D. farinae antigen 1 Der f l) and cat (cat antigen d1 Fel d 1) allergen and endotoxin content in settled house dust. Exposure to Der f 1 and Der p 1 plus Der f 1 >10 microg x g(-1) of mattress dust, respectively, increased the risk of wheeze and breathlessness (odds ratios (OR): 4.04, 95% confidence interval (CI): 1.53-10.64, OR: 2.78, 95% CI: 1.06-7.28). Fel d 1 >8 microg x g(-1) was positively associated with cough at night (OR: 2.74, 95%, CI: 1.22-.17), noteworthy also in the nonsensitized subjects. Subjects exposed to elevated concentrations of more than one allergen had an up to seven-fold increase in the risk of respiratory symptoms, compared to nonexposed subjects. Sensitized subjects exposed to elevated concentrations of Der f 1 or Fel d 1 were found to have the highest risk of asthma attacks and respiratory symptoms. No statistically significant association was found between exposure to endotoxins and respiratory health. Indoor exposure to Dermatophagoides farinae antigen 1 and cat antigen d1 is a risk factor for respiratory symptoms in adults, and for cat antigen d 1 even in nonsensitized subjects. The risk is increased if subjects are exposed to a mixture of allergens or if they are sensitized in addition to high exposure.

Adult↗
PubMed2001

Transcriptional activation by the human Hsp70-associating protein Hap50.

We investigated human Hap50, the large isoform of the previously characterized Hsp70/Hsc70-associating protein Hap46, also called BAG-1, for effects on transcriptional activities. Overproduction by transient transfection led to enhanced expression of reporter gene constructs in various cell types using different promoters, suggesting independence of promoter type. Similarly, overexpression of Hap50 resulted in increased levels of poly(A)(+ )mRNAs in HeLa, COS-7, 3T3 and HTC cells. Concomitantly, the expression of some selected endogenous genes, such as those coding for c-Jun and the glucocorticoid receptor, was enhanced significantly relative to actin. Nuclear runoff transcription assays using HeLa cells showed that the effect is caused by increased transcription rates rather than mRNA stabilization. Activation of transcription by Hap50 occurred at 37 degrees C and did not require prior thermal stress, as is the case for Hap46. In accordance with these biological effects, Hap50 is localized exclusively in the nuclear compartment of different cell types, whereas Hap46 is mostly cytoplasmic in unstressed cells, as revealed by use of fusion constructs with green fluorescent protein. High cellular levels of Hap50 were found to make cells less susceptible to adverse environmental effects such as heat stress. Our data suggest that Hap50 is a nuclear protein that acts in cells to increase the transcription of various genes.

3T3 Cells↗
PubMed2001

Beta(1-->3)-glucan in house dust of German homes: housing characteristics, occupant behavior, and relations with endotoxins, allergens, and molds.

beta(1-->3)-Glucans are potent proinflammatory agents that have been suggested to play a role in indoor-related respiratory health effects. The aim of this study was to assess whether beta(1-->3)-glucan concentrations in house dust are correlated with levels of endotoxins, allergens, and culturable mold spore counts in house dust. Further, the associations of beta(1-->3)-glucan with housing characteristics and occupant behavior were assessed. beta(1-->3)-Glucan was measured in settled house dust from living room floors of 395 homes of two German cities, Erfurt and Hamburg, with a specific enzyme immunoassay. Concentrations ranged from below the limit of detection to 19,013 microg/m(2) (22,588 microg/g dust). Concentrations per square meter were found to be correlated with endotoxins, mite and cat allergens, and culturable mold spores. Correlations were weaker when concentrations were expressed per gram of dust, indicating that variance in concentrations of all factors is largely determined by the amount of dust sampled. Associations between beta(1-->3)-glucan, housing characteristics, and occupant behavior were found for concentrations per square meter but not for concentrations per gram of dust. The following characteristics were associated with a significant increase in beta(1-->3)-glucan levels: carpets in the living room [means ratio (MR) = 1.9-2.1], keeping a dog inside (MR = 1.4), use of the home by four or more persons (MR = 1.4), use of the living room for > 180 hr/week (MR = 2.1), lower frequency of vacuum cleaning (MR = 1.6-3.0) and dust cleaning (MR = 1.2 and 1.4, respectively), and presence of mold spots during the past 12 months (MR = 1.4). We conclude that that the amount of dust sampled can be used as a proxy for hygiene and that beta(1-->3)-glucan concentrations per square meter are related to the amount of dust sampled.

Air Pollution, Indoor↗
PubMed2000

Dissection of the ATP-binding domain of the chaperone hsc70 for interaction with the cofactor Hap46.

Several unrelated proteins are known that specifically interact with members of the mammalian hsp70 chaperone protein family independent of the hsp70 substrate-binding site. One of these is Hap46, also called BAG-1, which binds to the ATP-binding domain of hsp70 and its constitutively expressed, highly homologous counterpart hsc70, thereby affecting nucleotide binding, as well as protein folding properties, of these molecular chaperones. In an attempt to delineate the potential contact sites on hsp70/hsc70 involved in this interaction we made use of the following two independent approaches: (i) screening of membrane-bound peptide libraries based on the sequence of the ATP-binding domain and (ii) the phage-display technique with random dodecapeptides. These approaches yielded partially overlapping results and identified several possible contact regions. On the space-filling model of hsc70, the two major contact areas for Hap46 delineated in the present study are located on the same side of the molecule on either subdomain that border the central cleft harboring the nucleotide-binding site. We suggest that this bridging affects the conformation of the ATP-binding domain in a way similar to the opening of the nucleotide-binding cleft produced in the bacterial hsp70 homologue DnaK upon binding its regulatory protein GrpE.

Adenosine Triphosphate↗
PubMed2000

Drafts in cold environments--the significance of air temperature and direction.

This paper concerns the influence of air temperature and of the direction of drafts on subjective and physiological responses. In three experimental series 58 healthy persons (50 men, 8 women, 20-29 yrs) were exposed to drafts in overall 446 experimental sessions. Drafts were applied either horizontally or vertically with mean air velocities of 0.1 to 0.3 m/s and a turbulence intensity of 50%. Air temperature was varied between 11 and 23 degrees C and metabolic rates between < 70 and 156 W/m2. These parameters were kept constant during the single one-hour sessions. The subjects were dressed for thermal neutrality. Draft-induced annoyance was registered every 5 minutes using a list of prescribed body parts and skin temperature was measured at the forearm and at the neck. Subjective and physiological responses were systematically related to air temperature. Draft-induced general annoyance, draft-induced local annoyance (neck, forearm) and the drop of the corresponding skin temperature were inversely related to air temperature. Concerning the direction horizontal drafts seem to cause somewhat stronger reactions. The predictive model developed by Toftum underestimates the percentage of persons annoyed. A modified version increases the predictive power significantly.

Adult↗
PubMed1999

The hsp70-associating protein Hap46 binds to DNA and stimulates transcription.

We investigated the ubiquitously expressed hsp70-associating protein Hap46, which is also called RAP46 and is homologous to BAG-1, for activities independent of hsp70 interactions. We observed in vitro binding to various DNA fragments but detected no apparent sequence specificity. Deletion of the amino-terminal decapeptide, which contains two clusters of three basic amino acids each, abolished the DNA-binding ability of Hap46. Similarly, exchange of either of these positively charged clusters for three alanines resulted in loss of DNA binding. Using a fusion of Hap46 and green fluorescent protein, we found preferential accumulation in cell nuclei on heat stress as compared with unstressed cells. The repressive effect of heat shock on overall transcriptional activity in human DU145 carcinoma cells was largely prevented when Hap46 was overexpressed by transfection. Such overproduction of Hap46 also resulted in enhanced expression of specific reporter gene constructs and in increased levels of mRNAs specific for hsp70 and hsp40 after temperature stress. In vitro transcription with nuclear extracts was stimulated greatly by Hap46. Like DNA binding, transcriptional enhancement required amino-terminally located basic amino acid residues but not the carboxyl-terminal portion of Hap46 known to participate in hsp70 interaction. Our results show that Hap46 is a bifunctional protein that can interact with both hsp70s and DNA, employing different portions of the molecule. They also suggest that Hap46 is involved in temperature-sensitive regulation of transcription, acting as a general transcriptional activator.

Amino Acid Sequence↗
PubMed1998

The chaperone cofactor Hop/p60 interacts with the cytosolic chaperonin-containing TCP-1 and affects its nucleotide exchange and protein folding activities.

The folding of protein structures often requires the presence of molecular chaperones and/or chaperonin complexes. We here investigated the inhibitory effects of the chaperone cofactors Hop/p60 and Hap46. By coimmunoprecipitation, we observed a direct interaction of the eukaryotic chaperonin-containing TCP-1 (CCT) purified from rabbit reticulocyte lysate with Hop/p60. By contrast, Hap46 was not coprecipitated. Binding of Hop/p60 to CCT is dependent on the presence of ATP or ADP and occurs through carboxyl-terminal sequences of Hop/p60. Hop/p60 significantly stimulates nucleotide exchange on CCT but not its ATPase activity, while Hap46 has no effects. We used denatured firefly luciferase as a model protein and found decreased binding to CCT in the presence of Hop/p60 and ATP. This coincides with the inhibitory effect of Hop/p60 on luciferase reactivation in an assay using purified CCT in combination with hsc70 and hsp40. We also observed that an antibody directed against one of the subunits of CCT efficiently inhibits refolding in a system which depends on crude reticulocyte lysate.

Adenosine Diphosphate↗
PubMed1998

RAP46 is a negative regulator of glucocorticoid receptor action and hormone-induced apoptosis.

RAP46 was first identified by its ability to bind the glucocorticoid receptor. It has since been reported to bind several cellular proteins, including the anti-apoptotic protein Bcl-2, but the biological significance of these interactions is unknown. Here we show that RAP46 binds the hinge region of the glucocorticoid receptor and inhibits DNA binding and transactivation by the receptor. We further show that overexpression of RAP46 in mouse thymoma S49.1 cells inhibits glucocorticoid-induced apoptosis. Conversely, glucocorticoid-induced apoptosis and transactivation were enhanced after treating S49.1 cells with the immunosuppressant rapamycin, which down-regulates cellular levels of BAG-1, the mouse homolog of RAP46. The effect of rapamycin can, however, be overcome by overexpression of RAP46. These results together identify RAP46 as a protein that controls glucocorticoid-induced apoptosis through its negative regulatory action on the transactivation property of the glucocorticoid receptor.

Animals↗