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PubMed2001

Dwarf8 polymorphisms associate with variation in flowering time.

Historically, association tests have been used extensively in medical genetics, but have had virtually no application in plant genetics. One obstacle to their application is the structured populations often found in crop plants, which may lead to nonfunctional, spurious associations. In this study, statistical methods to account for population structure were extended for use with quantitative variation and applied to our evaluation of maize flowering time. Mutagenesis and quantitative trait locus (QTL) studies suggested that the maize gene Dwarf8 might affect the quantitative variation of maize flowering time and plant height. The wheat orthologs of this gene contributed to the increased yields seen in the 'Green Revolution' varieties. We used association approaches to evaluate Dwarf8 sequence polymorphisms from 92 maize inbred lines. Population structure was estimated using a Bayesian analysis of 141 simple sequence repeat (SSR) loci. Our results indicate that a suite of polymorphisms associate with differences in flowering time, which include a deletion that may alter a key domain in the coding region. The distribution of nonsynonymous polymorphisms suggests that Dwarf8 has been a target of selection.

Genes, Plant↗

PubMed2003

Genetic and maternal effect influences on viability of common frog tadpoles under different environmental conditions.

The influence of environmental stress on the expression of genetic and maternal effects on the viability traits has seldom been assessed in wild vertebrates. We have estimated genetic and maternal effects on the viability (viz probability of survival, probability of being deformed, and body size and shape) of common frog, Rana temporaria, tadpoles under stressful (low pH) and nonstressful (neutral pH) environmental conditions. A Bayesian analysis using generalized linear mixed models was applied to data from a factorial laboratory experiment. The expression of additive genetic variance was independent of pH treatments, and all traits were significantly heritable (survival: h2 approximately 0.08; deformities: h2 approximately 0.26; body size: h2 approximately 0.12; body shape: h2 approximately 0.14). Likewise, nonadditive genetic contributions to variation in all traits were significant, independent of pH treatments and typically of magnitude similar to the additive genetic effects. Maternal effects were large for all traits, especially for viability itself, and their expression was partly dependent on the environment. In the case of body size, the maternal effects were mediated largely through egg size. In general, the results give little evidence for the conjecture that environmental stress created by low pH would impact strongly on the genetic architecture of fitness-related traits in frogs, and hamper adaptation to stress caused by acidification. The low heritabilities and high dominance contributions conform to the pattern typical for traits subject to relatively strong directional selection.

Analysis of Variance↗

PubMed2006

Spatiotemporal structure of genetic variation of a spreading plant metapopulation on dynamic riverbanks along the Meuse River.

Long-distance seed dispersal is a crucial determinant of within-population genetic variability and among-population genetic differentiation in plant metapopulations undergoing recurrent local extinctions and (re-)colonization. We investigated the spatial and temporal structure of genetic variation in a metapopulation of Sisymbrium austriacum located along a dynamic river system using dominant AFLP markers. Data on riverbank dynamics and colonization history allowed separating populations based on their age (< or =5 vs >5 years old). Bayesian analysis of population genetic structure indicated that populations were significantly differentiated from each other, but Mantel tests revealed that there was no relationship between pairwise geographic and genetic distances, suggesting that long-distance seed dispersal partly determines spatial genetic structure. Recent populations were less differentiated from each other than old populations. Analysis of molecular variance (AMOVA) indicated that both spatial factors and population age significantly determined genetic diversity, the effects of age being more important than spatial location. Clustering analysis revealed five large clusters, which were related primarily to population age and to a minor extent to geographical location. Our results indicate that the recurrent formation and destruction of riverbank habitats following peak flow events have a large impact on genetic diversity of riparian plant species.

Belgium↗

PubMed2006

Individual-based assessment of population structure and admixture in Austrian, Croatian and German draught horses.

All over Europe, the number of draught horses has decreased drastically during the last 50 years. As a prerequisite for efficient management decisions, we analysed the conservation status in Austrian (Noriker Carinthia - NC, Noriker Salzburg - NS), Croatian (Croatian Coldblood - C, Posavina horse - P) and German (Altmaerkisch Coldblood - A, Black Forest horse - BF, Mecklenburg Coldblood - M, Rhenish German Draught horse - R, Saxon Thuringa Coldblood - ST, Schleswig Draught horse - Sch, South German Coldblood - SG) draught horses (434) using multilocus genotypic information from 30 (effectively 27) microsatellite loci. Populations located in areas with less intensive agricultural production (C, NC, NS, P and SG) had greater diversity within the population and estimated effective population size than A, BF, Sch, M, R and ST populations. The PCA plots revealed that populations form five separate groups. The 'Noriker' group (NC, NS and SG) and the 'Rhenish' group (A, M, R and ST) were the most distinctive (pairwise F(ST) values ranged from 0.078 to 0.094). The 'Croatian' group (C and P) was in the centre, while the BF and Sch populations formed two out-groups. A posterior Bayesian analysis detected further differentiation, mainly caused by political and geographical factors. Thus, it was possible to separate the South German Coldblood from the Austrian Noriker population where no subpopulation structure was detected. The admixture analysis revealed imprecise classification between C and P populations. A small but notable separation of R from A, M and ST populations was detected, while Sch and BF populations remained as out-groups. The information obtained should aid in making efficient conservation decisions.

Animals↗

PubMed1999

A pharmacokinetic model for alpha interferon administered subcutaneously.

AIMS: To model the pharmacokinetic profiles of alpha interferon (alphaIFN) after a single subcutaneous (s.c.) injection of 3 million units of alpha 2b interferon, to correlate the pharmacokinetic parameters with patient demographic covariates, and to develop a limiting sampling strategy for determining the alphaIFN plasma area under the curve of concentration vs time (AUC). METHODS: The plasma alphaIFN pharmacokinetics were determined in 27 patients with chronic hepatitis C virus infection after the first s.c. injection of the drug. Ten patients had normal renal function and 17 were chronic haemodialysis patients. Plasma samples were assayed by an Elisa method. Concentration-time data was analysed by a population approach using NONMEM. RESULTS: The pharmacokinetic model which better described the concentration vs time data was a one-compartment model with two processes of absorption: a zero-order followed by a first-order process. The mean clearance of dialysis patients represented 37% (with 95% confidence interval: 30% -44%) of the mean value of the patients with normal renal function. The volume of distribution was significantly correlated to the body surface area. Bayesian analysis using NONMEM allowed determination of the individual plasma AUC from three samples within the 24 h period post s.c. injection. CONCLUSIONS: The present pharmacokinetic model will allow one to obtain individual parameters such as, the area under the curve of concentration vs time from a limited-sampling strategy, and to perform pharmacokinetic-pharmacodynamic analysis of combined alphaIFN plasma concentrations and viraemic data.

Adult↗

PubMed2001

Three quantitative approaches to the diagnosis of abdominal pain in children: practical applications of decision theory.

BACKGROUND/PURPOSE: The authors compared 3 quantitative methods for assisting clinicians in the differential diagnosis of abdominal pain in children, where the most common important endpoint is whether the patient has appendicitis. Pretest probability in different age and sex groups were determined to perform Bayesian analysis, binary logistic regression was used to determine which variables were statistically significantly likely to contribute to a diagnosis, and recursive partitioning was used to build decision trees with quantitative endpoints. METHODS: The records of all children (1,208) seen at a large urban emergency department (ED) with a chief complaint of abdominal pain were immediately reviewed retrospectively (24 to 72 hours after the encounter). Attempts were made to contact all the patients' families to determine an accurate final diagnosis. A total of 1,008 (83%) families were contacted. Data were analyzed by calculation of the posttest probability, recursive partitioning, and binary logistic regression. RESULTS: In all groups the most common diagnosis was abdominal pain (ICD-9 Code 789). After this, however, the order of the most common final diagnoses for abdominal pain varied significantly. The entire group had a pretest probability of appendicitis of 0.06. This varied with age and sex from 0.02 in boys 2 to 5 years old to 0.16 in boys older than 12 years. In boys age 5 to 12, recursive partitioning and binary logistic regression agreed on guarding and anorexia as important variables. Guarding and tenderness were important in girls age 5 to 12. In boys age greater than 12, both agreed on guarding and anorexia. Using sensitivities and specificities from the literature, computed tomography improved the posttest probability for the group from.06 to.33; ultrasound improved it from.06 to.48; and barium enema improved it from.06 to.58. CONCLUSIONS: Knowing the pretest probabilities in a specific population allows the physician to evaluate the likely diagnoses first. Other quantitative methods can help judge how much importance a certain criterion should have in the decision making and how much a particular test is likely to influence the probability of a correct diagnosis. It now should be possible to make these sophisticated quantitative methods readily available to clinicians via the computer.

Abdominal Pain↗

PubMedDate not supplied

Principles for combining radionuclide and echocardiographic stress imaging into a single clinical laboratory service.

Physicians who supervise stress-imaging procedures, whether they be stress nuclear or stress echocardiography tests, are increasingly required to incorporate important principles in the interpretation of the test results. Important guiding principles, common to these tests, are reviewed in this article. The principles include the following: avoiding categorical interpretation of test results; identifying equivocal test responses; combining diagnostic and prognostic information in rendering information to referring physicians; integrating the stress test results with other clinical data and using Bayesian analysis as a decision-aid; reporting one's confidence in the interpretation of the results of testing; characterizing both the extent and severity of any induced myocardial ischemia; and analyzing the test results in a blinded fashion, without prior knowledge of patient clinical history

Bayes Theorem↗

PubMed2002

The pulmonologist's perspective regarding the solitary pulmonary nodule.

The pulmonologist's goal in managing a patient with a solitary pulmonary nodule is to distinguish the benign from malignant nodule and, where malignancy is either confirmed or strongly suspected, to expedite resection. By using established clinical features (eg, age, smoking status) and radiographic findings (eg, calcification, growth rate, size), a probability of malignancy can be determined. If necessary, noninvasive or adjuvant invasive testing is used to alter the probability to one that permits observation or demands resection. The proper use of these tests mandates knowledge about their performance characteristics. Decision-analytic approaches, using Bayesian analysis, may assist with the calculation of probability. These models have not consistently outperformed the clinician or adjuvant testing. The use of low-dose computed tomography (CT) scanning as a screening tool has led to the discovery of many small, indeterminate nodules. Management decisions for these nodules are influenced by their low prevalence of malignancy and small size. Future advances will add to our ability to effectively meet our stated goal.

Decision Making↗

PubMed2002

Relationship between neonatal leukemoid reaction and bronchopulmonary dysplasia in low-birth-weight infants: a cross-sectional study.

Leukemoid reaction in low-birth-weight (LBW) infants is a rare, recently documented phenomenon, implicated in the sequence of multiorgan inflammatory diseases of preterm infants. The aim of the present paper is to establish whether a neonatal leukemoid reaction is related to bronchopulmonary dysplasia (BPD) development in LBW infants. The design was a case-controlled, retrospective study of all premature infants (born </=31 weeks' gestation) admitted to the neonatal intensive care unit (NICU) over a period of 3 years, from January 1998 to December 2000. The infants who developed BPD formed the study group, while the remainder without pulmonary sequelae, matched for gestational age formed the control group. Leukemoid reaction was considered a white blood cell (WBC) count >40,000/mm 3. The relation between BPD occurrence and WBC counts was studied by Bayesian analysis, dividing WBC counts in 5 progressive bands of 10,000 WBC/mm 3, starting from <10,000 to >40,000/mm 3. Five of 50 BPD infants studied demonstrated WBC counts >40,000/mm 3, with an incidence of 10%, while no control preterm infants presented neonatal leukemoid reaction; the estimated number difference is statistically significant ( p <0.001). There was no other significant association demonstrated between maternal or neonatal variables and leukemoid reaction, including chorioamnionitis, sepsis, and the use of antenatal steroids. Our findings provide further data for the identification of prematures exposed to pro-inflammatory cytokines in utero; the injury responsible for BPD in a subset of prematures may begin with a transient leukemoid reaction.

Bronchopulmonary Dysplasia↗

PubMed2000

[The walking-counting test. A simple test for assessing the risk of falling].

BACKGROUND AND OBJECTIVE: Falls and fractures caused by falls are common in the elderly. Interventions to prevent such falls and/or fractures are available. But there are no appropriate tests in general practice for the targeted use of preventative measures. This prospective study was undertaken to validate a simple "walking and counting" test for assessing the risk of falling. PATIENTS AND METHODS: 92 patients of a general practice, aged between 70 and 96 years (64 women, 28 men) were included in the study. All patients were given the "walking and counting" test in which they would walk as quickly as possible along a stretch of 4 m, repeated immediately in a 3-step sequence while counting backwards from 100. The walking speed without counting was measured initially (vo), as was the speed during distraction by counting (vD). The difference between the two values, delta v%, measured the percentage change under distraction. In 80 patients it was possible to record the number of falls over the entire subsequent period of 12 months. All data were assessed by relative operating characteristics (ROC) and Bayesian analysis. RESULTS: 40 patients had falls in the follow-up period of 12 months, falls being more frequent the higher the age and if there had been previous falls. While all patients were able to do the test, individual values differed considerably. There was a statistical correlation between the calculated change in walking speed and the risk of falling. Using a threshold value of 20% slowing of the walking speed, the test sensitivity was 95% (0.51-0.81: 95% confidence interval) and the specificity 0.85 (0.71-0.93), with a positive predictive value of 0.77 (0.59-0.84). CONCLUSION: The walking and counting test is a simple and informative test, more reliable than other parameters, to predict for an individual patient with danger of falling over a 12-months period. It provides the general practitioner with a tool for demanding appropriately targeted preventative measures.

Accidental Falls↗

PubMed2002

Potential applications and pitfalls of Bayesian inference of phylogeny.

Only recently has Bayesian inference of phylogeny been proposed. The method is now a practical alternative to the other methods; indeed, the method appears to possess advantages over the other methods in terms of ability to use complex models of evolution, ease of interpretation of the results, and computational efficiency. However, the method should be used cautiously. The results of a Bayesian analysis should be examined with respect to the sensitivity of the results to the priors used and the reliability of the Markov chain Monte Carlo approximation of the probabilities of trees.

Bayes Theorem↗

PubMed2004

Molecular phylogeny, historical biogeography, and divergence time estimates for swallowtail butterflies of the genus Papilio (Lepidoptera: Papilionidae).

Swallowtail butterflies are recognized as model organisms in ecology, evolutionary biology, genetics, and conservation biology but present numerous unresolved phylogenetic problems. We inferred phylogenetic relationships for 51 of about 205 species of the genus Papilio (sensu lato) from 3.3-Kilobase (kb) sequences of mitochondrial and nuclear DNA (2.3 kb of cytochrome oxidases I and II and 1.0 kb of elongation factor 1 alpha). Congruent phylogenetic trees were recovered within Papilio from analyses of combined data using maximum likelihood, Bayesian analysis, and maximum parsimony bootstrap consensus. Several disagreements with the traditional classification of Papilio were found. Five major previously hypothesized subdivisions within Papilio were well supported: Heraclides, Pterourus, Chilasa, Papilio (sensu stricto), and Eleppone. Further studies are required to clarify relationships within traditional "Princeps," which was paraphyletic. Several biologically interesting characteristics of Papilio appear to have polyphyletic origins, including mimetic adults, larval host associations, and larval morphology. Early diversification within Papilio is estimated at 55-65 million years ago based on a combination of biogeographic time constraints rather than fossils. This divergence time suggests that Papilio has slower apparent substitution rates than do Drosophila and fig-pollinating wasps and/or divergences corrected using best-fit substitution models are still being consistently underestimated. The amount of sequence divergence between Papilio subdivisions is equivalent to divergences between genera in other tribes of the Papilionidae, and between genera of moths of the noctuid subfamily Heliothinae.

Animals↗

PubMed2004

Testing the phylogenetic position of a parasitic plant (Cuscuta, Convolvulaceae, asteridae): Bayesian inference and the parametric bootstrap on data drawn from three genomes.

Previous findings on structural rearrangements in the chloroplast genome of Cuscuta (dodder), the only parasitic genus in the morning-glory family, Convolvulaceae, were attributed to its parasitic life style, but without proper comparison to related nonparasitic members of the family. Before molecular evolutionary questions regarding genome evolution can be answered, the phylogenetic problems within the family need to be resolved. However, the phylogenetic position of parasitic angiosperms and their precise relationship to nonparasitic relatives are difficult to infer. Problems are encountered with both morphological and molecular evidence. Molecular data have been used in numerous studies to elucidate relationships of parasitic taxa, despite accelerated rates of sequence evolution. To address the question of the position of the genus Cuscuta within Convolvulaceae, we generated a new molecular data set consisting of mitochondrial (atpA) and nuclear (RPB2) genes, and analyzed these data together with an existing chloroplast data matrix (rbcL, atpB, trnL-F, and psbE-J), to which an additional chloroplast gene (rpl2) was added. This data set was analyzed with an array of phylogenetic methods, including Bayesian analysis, maximum likelihood, and maximum parsimony. Further exploration of data was done by using methods of phylogeny hypothesis testing. At least two nonparasitic lineages are shown to diverge within the Convolvulaceae before Cuscuta. However, the exact sister group of Cuscuta could not be ascertained, even though many alternatives were rejected with confidence. Caution is therefore warranted when interpreting the causes of molecular evolution in Cuscuta. Detailed comparisons with nonparasitic Convolvulaceae are necessary before firm conclusions can be reached regarding the effects of the parasitic mode of life on patterns of molecular evolution in Cuscuta.

Base Sequence↗

PubMed2004

Biogeography of the Pistia clade (Araceae): based on chloroplast and mitochondrial DNA sequences and Bayesian divergence time inference.

Pistia stratiotes (water lettuce) and Lemna (duckweeds) are the only free-floating aquatic Araceae. The geographic origin and phylogenetic placement of these unrelated aroids present long-standing problems because of their highly modified reproductive structures and wide geographical distributions. We sampled chloroplast (trnL-trnF and rpl20-rps12 spacers, trnL intron) and mitochondrial sequences (nad1 b/c intron) for all genera implicated as close relatives of Pistia by morphological, restriction site, and sequencing data, and present a hypothesis about its geographic origin based on the consensus of trees obtained from the combined data, using Bayesian, maximum likelihood, parsimony, and distance analyses. Of the 14 genera closest to Pistia, only Alocasia, Arisaema, and Typhonium are species-rich, and the latter two were studied previously, facilitating the choice of representatives that span the roots of these genera. Results indicate that Pistia and the Seychelles endemic Protarum sechellarum are the basalmost branches in a grade comprising the tribes Colocasieae (Ariopsis, Steudnera, Remusatia, Alocasia, Colocasia), Arisaemateae (Arisaema, Pinellia), and Areae (Arum, Biarum, Dracunculus, Eminium, Helicodiceros, Theriophonum, Typhonium). Unexpectedly, all Areae genera are embedded in Typhonium, which throws new light on the geographic history of Areae. A Bayesian analysis of divergence times that explores the effects of multiple fossil and geological calibration points indicates that the Pistia lineage is 90 to 76 million years (my) old. The oldest fossils of the Pistia clade, though not Pistia itself, are 45-my-old leaves from Germany; the closest outgroup, Peltandreae (comprising a few species in Florida, the Mediterranean, and Madagascar), is known from 60-my-old leaves from Europe, Kazakhstan, North Dakota, and Tennessee. Based on the geographic ranges of close relatives, Pistia likely originated in the Tethys region, with Protarum then surviving on the Seychelles, which became isolated from Madagascar and India in the Late Cretaceous (85 my ago). Pistia and Protarum provide striking examples of ancient lineages that appear to have survived in unique or isolated habitats.

Araceae↗

PubMed2003

The effect of metformin on ovarian stimulation and in vitro fertilization in insulin-resistant women with polycystic ovary syndrome: an open-label randomized cross-over trial.

Metformin effectively restores insulin sensitivity in insulin-resistant women with polycystic ovary syndrome (PCOS). We examined whether metformin, given prior to and during ovarian stimulation for in vitro fertilization (IVF), altered follicle stimulating hormone (FSH) requirement and increased the number of collected oocytes in these women. Seventeen insulin-resistant women with PCOS were recruited to our IVF unit to receive two consecutive cycles of ovarian stimulation with or without metformin co-treatment, the order of treatments being randomized using a table of random numbers. Metformin treatment (1500 mg/day) started 3 weeks before downregulation with buserelin acetate and was continued throughout ovarian stimulation with human recombinant FSH. Nine women completed both cycles, the results of eight women being excluded because of pregnancy after the first cycle (n = 4) or because the protocol of the study was not followed (n = 4). Mean total FSH dose was 2301 IU (range 1500-6563 IU) in metformin cycles and 2174 IU (range 1200-3900 IU) in parallel control cycles, while the mean number of collected oocytes was 8.6 (range 2-28) and 4.6 (range 1-16), respectively. Bayesian analysis showed probabilities of 0.05 that metformin reduces FSH requirement by at least 10%, and of 0.61 that at least 10% more oocytes are collected after metformin co-treatment. Co-administration of metformin is therefore likely to increase the number of oocytes collected after ovarian stimulation in insulin-resistant women with PCOS but is unlikely to reduce the requirement for FSH.

Adult↗

PubMed2005

Identification of QTL for production traits in chickens.

If the poultry industry hopes to continue to flourish, the identification of potential quantitative trait loci (QTL) for production-related traits must be pursued This remains true despite the sequencing of the chicken genome. In view of this need, a scan of the chicken genome using 72 microsatellite markers was carried out on a meat-type x egg-type resource population measured for production and egg quality traits. Using a Bayesian analysis, potential QTL for a number of traits were identified on several chromosomes. Evidence of eight QTL regions associated with a total of eight traits (specific gravity, albumin height, Haugh score, shell shape, total number of eggs, final body weight, gain, and feed efficiency) was found. Two of these regions, one spanning the area of 263/287 cM on GAA01 and the other spanning the area of 23/28 cM on GAA02, were associated with multiple QTL.

Animals↗

PubMed1999

A reanalysis of IgM Western blot criteria for the diagnosis of early Lyme disease.

A two-step approach for diagnosis of Lyme disease, consisting of an initial EIA followed by a confirmatory Western immunoblot, has been advised by the Centers for Disease Control and Prevention (CDC). However, these criteria do not examine the influence of the prior probability of Lyme disease in a given patient on the predictive value of the tests. By using Bayesian analysis, a mathematical algorithm is proposed that computes the probability that a given patient's Western blot result represents Lyme disease. Assuming prior probabilities of early Lyme disease of 1%-10%, the current CDC minimum criteria for IgM immunoblot interpretation yield posttest probabilities of 4%-32%. The value of the two-step approach for diagnosis of early Lyme disease may be limited in populations at lower risk of disease or when patients present with atypical signs and symptoms.

Blotting, Western↗

PubMed1998

Can biological activity be maintained at ultra-high dilution? An overview of homeopathy, evidence, and Bayesian philosophy.

Forty percent of general practitioners in the Netherlands practice homeopathy. With over 100 homeopathic medical schools, homeopathy is practiced in India along with conventional Western medicine in government clinics. In Britain, 42% of general practitioners refer patients to homeopaths. Two recent meta-analyses of homeopathy both indicate that there is enough evidence to show that homeopathy has added effects over placebo. Against this evidence is a backdrop of considerable scientific scepticism. Homeopathic remedies are diluted substances--some are so diluted that statistically there are no molecules present to explain their proposed biological effects (ultra-high dilutions or UHDs). Without knowledge of the evidence, most scientists would reject UHD effects because of their intrinsic implausibility in the light of our current scientific understanding. The objective of this article is to critically review the major pieces of evidence on UHD effects and suggest how the scientific community should respond to its challenge. Such evidence has been conducted on a diverse range of assays--immunologic, physiological, behavioral, biochemical, and clinical in the form of trials of homeopathic remedies. Evidence of UHD effects has attracted the attention of physicists who have speculated on their physical mechanisms. Included is a critique of several experiments that form the Benveniste affair which was sparked by a publication in Nature that advocated the existence of UHD effects of anti-immunoglobulin E (IgE) on human basophils, and is the paradigm example of how a controversial phenomenon can split the scientific community. It is argued that if the phenomenon was uncontroversial, the evidence suffices to show that UHD effects exist. However, given that the observations contradict well-established theory, normal science has to be abandoned and scientists need to decide for themselves what the likelihood of UHD effects are. Bayesian analysis describes how scientists ought rationally to change their prior beliefs in the light of evidence. Theories by Kuhn and Lakatos indicate that whether UHD effects are proved or not depends on the beliefs and behaviors of scientists in their communities. This article argues that there is as yet insufficient evidence to drive rational scientists to a consensus over UHD effects, even if they possessed knowledge of all the evidence. The difficulty in publishing high-quality UHD research in conventional journals prevents a fair assessment of UHD effects. Given that the existence of UHD effects would revolutionize science and medicine, and given the considerable empirical evidence of them, the philosophies of science tell us that possible UHD effects warrant serious investigation by conventional scientists and serious attention by scientific journals.

Animals↗