Innervation of putative rapidly adapting mechanoreceptors by calbindin- and calretinin-immunoreactive primary sensory neurons in the rat.

Calbindin and calretinin are two homologous calcium-binding proteins that are expressed by subpopulations of primary sensory neurons. In the present work, we have studied the distribution of the neurons expressing calbindin and calretinin in dorsal root ganglia of the rat and their peripheral projections. Calbindin and calretinin immunoreactivities were expressed by subpopulations of large- and small-sized primary sensory neurons and colocalized in a majority of large-sized ones. The axons emerging from calbindin- or calretinin-immunoreactive neurons innervated muscle spindles, Pacini corpuscles and subepidermal lamellar corpuscles in the glabrous skin, formed palisades of lanceolate endings around hairs and vibrissae, and gave rise to intraepidermal nerve endings in the digital skin. Since most of these afferents are considered as rapidly adapting mechanoreceptors, it is concluded that calbindin- or calretinin-expressing neurons innervate particular mechanoreceptors that display physiological characteristics of rapid adaptation to stimuli.

Erosivity of rainfall in Lages, Santa Catarina, Brazil

The erosive capacity of rainfall can be expressed by an index and knowing it allows recommendation of soil management and conservation practices to reduce water erosion. The objective of this study was to calculate various indices of rainfall erosivity in Lages, Santa Catarina, Brazil, identify the best one, and discover its temporal distribution. The study was conducted at the Center of Agricultural and Veterinary Sciences, Lages, Santa Catarina, using daily rainfall charts from 1989 to 2012. Using the computer program Chuveros , 107 erosivity indices were obtained, which were based on maximum intensity in 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 110, 120, 135, 150, 165, 180, 210, and 240 min of duration and on the combination of these intensities with the kinetic energy obtained by the equations of Brown & Foster, Wagner & Massambani, and Wischmeier & Smith. The indices of the time period from 1993 to 2012 were correlated with the respective soil losses from the standard plot of the Universal Soil Loss Equation (USLE) in order to select the erosivity index for the region. Erosive rainfall accounted for 83 % of the mean annual total volume of 1,533 mm. The erosivity index (R factor) of rainfall recommended for Lages is the EI30, whose mean annual value is 5,033 MJ mm ha-1 h-1, and of this value, 66 % occurs from September to February. Mean annual erosivity has a return period estimated at two years with a 50 % probability of occurrence.

MORPHOLOGICAL AND MICROMORPHOLOGICAL CHANGES IN THE STRUCTURE OF A RHODIC HAPLUDOX AS A RESULT OF AGRICULTURAL MANAGEMENT

In evaluation of soil quality for agricultural use, soil structure is one of the most important properties, which is influenced not only by climate, biological activity, and management practices but also by mechanical and physico-chemical forces acting in the soil. The purpose of this study was to evaluate the influence of conventional agricultural management on the structure and microstructure of a Latossolo Vermelho distroférrico típico (Rhodic Hapludox) in an experimental area planted to maize. Soil morphology was described using the crop profile method by identifying the distinct structural volumes called Morphologically Homogeneous Units (MHUs). For comparison, we also described a profile in an adjacent area without agricultural use and under natural regrowth referred to as Memory. We took undisturbed samples from the main MHUs so as to form thin sections and blocks of soil for micromorphological and micromorphometrical analyses. Results from the application of the crop profile method showed the occurrence of the following structural types: loose (L), fragmented (F) and continuous (C) in both profiles analyzed. In the Memory soil profile, the fragmented structures were classified as Fptμ∆+tf and Fmt∆μ, whose micromorphology shows an enaulic-porphyric (porous) relative distribution with a great deal of biological activity as indicated by the presence of vughs and channels. Lower down, from 0.20 to 0.35 m, there was a continuous soil volume (sub-type C∆μ), with a subangular block microstructure and an enaulic-porphyric relative distribution, though in this case more compact and with aggregate coalescence and less biological activity. The micromorphometrical study of the soil of the Memory Plot showed the predominance of complex pores in NAM (15.03 %), Fmt∆μ (11.72 %), and Fptμ∆+tf (7.73 %), and rounded pores in C∆μ (8.21 %). In the soil under conventional agricultural management, we observed fragmented structures similar to the Memory Plot from 0.02 to 0.20 m, followed by a volume with a compact continuous structure (C∆μ), without visible porosity and with few roots. In the MHUs under conventional management, reduction in the packing pores (40 %) was observed, mainly in the continuous units (C). The microstructure had well-defined blocks, with the occurrence of planar pores and less evidence of biological activity. In conclusion, the morphological and micromorphological analyses of the soil profiles studied offered complementary information regarding soil structural quality, especially concerning the changes in pore types as result of mechanical stress undergone by the soil.

EFFECT OF SOIL TILLAGE AND PLANT RESIDUE ON SURFACE ROUGHNESS OF AN OXISOL UNDER SIMULATED RAIN

Surface roughness of the soil is formed by mechanical tillage and is also influenced by the kind and amount of plant residue, among other factors. Its persistence over time mainly depends on the fundamental characteristics of rain and soil type. However, few studies have been developed to evaluate these factors in Latossolos (Oxisols). In this study, we evaluated the effect of soil tillage and of amounts of plant residue on surface roughness of an Oxisol under simulated rain. Treatments consisted of the combination of the tillage systems of no-tillage (NT), conventional tillage (CT), and minimum tillage (MT) with rates of plant residue of 0, 1, and 2 Mg ha-1 of oats (Avena strigosa Schreb) and 0, 3, and 6 Mg ha-1 of maize (Zea mays L.). Seven simulated rains were applied on each experimental plot, with intensity of 60±2 mm h-1 and duration of 1 h at weekly intervals. The values of the random roughness index ranged from 2.94 to 17.71 mm in oats, and from 5.91 to 20.37 mm in maize, showing that CT and MT are effective in increasing soil surface roughness. It was seen that soil tillage operations carried out with the chisel plow and the leveling disk harrow are more effective in increasing soil roughness than those carried out with the heavy disk harrow and leveling disk harrow. The roughness index of the soil surface decreases exponentially with the increase in the rainfall volume applied under conditions of no tillage without soil cover, conventional tillage, and minimum tillage. The oat and maize crop residue present on the soil surface is effective in maintaining the roughness of the soil surface under no-tillage.

Hirsutisme: que faire [Hirsutism, what can be done?]

Hirsutism is a relatively frequent condition in an ambulatory setting affecting about 4% of women. A rational clinical and biochemical diagnostic approach assures an optimal treatment directed at etiologic and pathogenetic factors. The diagnosis of hyperandrogenism is evaluated considering the pathophysiologic mechanisms responsible for excessive growth of hair. Ovarian and adrenal tumors are the most serious diseases that have to be excluded by clinical and biochemical tests. The other causes for hirsutism are treatable by a great variety of modalities, available drugs can inhibit pituitary gonadotropins, the hypothalamo-pituitary axis and the conversion of testosterone into biologically active substrate. Finally the binding of androgens to its receptor can be blocked. These possibilities for treatment and their indications for the different etiologies of hirsutism are discussed.

NITROGEN LOSS BY EROSION FROM MECHANICALLY TILLED AND UNTILLED SOIL UNDER SUCCESSIVE SIMULATED RAINFALLS

The description of the fate of fertilizer-derived nitrogen (N) in agricultural systems is an essential tool to enhance management practices that maximize nutrient use by crops and minimize losses. Soil erosion causes loss of nutrients such as N, causing negative effects on surface and ground water quality, aside from losses in agricultural productivity by soil depletion. Studies correlating the percentage of fertilizer-derived N (FDN) with soil erosion rates and the factors involved in this process are scarce. The losses of soil and fertilizer-derived N by water erosion in soil under conventional tillage and no tillage under different rainfall intensities were quantified, identifying the intervening factors that increase loss. The experiment was carried out on plots (3.5 × 11 m) with two treatments and three replications, under simulated rainfall. The treatments consisted of soil with and soil without tillage. Three successive rainfalls were applied in intervals of 24 h, at intensities of 30 mm/h, 30 mm/h and 70 mm/h. The applied N fertilizer was isotopically labeled (15N) and incorporated into the soil in a line perpendicular to the plot length. Tillage absence resulted in higher soil losses and higher total nitrogen losses (TN) by erosion induced by the rainfalls. The FDN losses followed another pattern, since FDN contributions were highest from tilled plots, even when soil and TN losses were lowest, i.e., the smaller the amount of eroded sediment, the greater the percentage of FDN associated with these. Rain intensity did not affect the FDN loss, and losses were greatest after less intense rainfalls in both treatments.

Liming in Agricultural Production Models with and Without the Adoption of Crop-Livestock Integration

ABSTRACT Perennial forage crops used in crop-livestock integration (CLI) are able to accumulate large amounts of straw on the soil surface in no-tillage system (NTS). In addition, they can potentially produce large amounts of soluble organic compounds that help improving the efficiency of liming in the subsurface, which favors root growth, thus reducing the risks of loss in yield during dry spells and the harmful effects of “overliming”. The aim of this study was to test the effects of liming on two models of agricultural production, with and without crop-livestock integration, for 2 years. Thus, an experiment was conducted in a Latossolo Vermelho (Oxisol) with a very clayey texture located in an agricultural area under the NTS in Bandeirantes, PR, Brazil. Liming was performed to increase base saturation (V) to 65, 75, and 90 % while one plot per block was maintained without the application of lime (control). A randomized block experimental design was adopted arranged in split-plots and four plots/block, with four replications. The soil properties evaluated were: pH in CaCl2, soil organic matter (SOM), Ca, Mg, K, Al, and P. The effects of liming were observed to a greater depth and for a long period through mobilization of ions in the soil, leading to a reduction in SOM and Al concentration and an increase in pH and the levels of Ca and Mg. In the first crop year, adoption of CLI led to an increase in the levels of K and Mg and a reduction in the levels of SOM; however, in the second crop year, the rate of decline of SOM decreased compared to the decline observed in the first crop year, and the level of K increased, whereas that of P decreased. The extent of the effects of liming in terms of depth and improvement in the root environment from the treatments were observed only partially from the changes observed in the chemical properties studied.

Sheep Excreta as Source of Nitrous Oxide in Ryegrass Pasture in Southern Brazil

ABSTRACT Livestock urine and dung are important components of the N cycle in pastures, but little information on its effect on soil nitrous oxide (N2O) emissions is available. We conducted a short-term (39-day) trial to quantify the direct N2O-N emissions from sheep excreta on an experimental area of ryegrass pasture growing on a Typic Paleudult in southern Brazil. Four rates of urine-N (161, 242, 323, and 403 kg ha-1 N) and one of dung-N (13 kg ha-1 N) were applied, as well as a control plot receiving no excreta. The N2O-N emission factor (EF = % of added N released as N2O-N) for urine and dung was calculated, taking into account the N2O fluxes in the field, over a period of 39 days. The EF value of the urine and dung was used to estimate the emissions of N2O-N over a 90-day period of pasture in the winter under two grazing intensities (2.5 or 5.0 times the herbage intake potential of grazing lambs). The soil N2O-N fluxes ranged from 4 to 353 µg m-2h-1. The highest N2O-N fluxes occurred 16 days after application of urine and dung, when the highest soil nitrate content was also recorded and the water-filled pore space exceeded 60 %. The mean EF for urine was 0.25 % of applied N, much higher than that for dung (0.06 %). We found that N2O-N emissions for the 90-day winter pasture period were 0.54 kg ha-1 for low grazing intensity and 0.62 kg ha-1 for moderate grazing intensity. Comparison of the two forms of excreta show that urine was the main contributor to N2O-N emissions (mean of 36 %), whereas dung was responsible for less than 0.1 % of total soil N2O-N emissions.

MMP13 mutations are the cause of recessive metaphyseal dysplasia, Spahr type.

Metaphyseal dysplasia, Spahr type (MDST; OMIM 250400) was described in 1961 based on the observation of four children in one family who had rickets-like metaphyseal changes but normal blood chemistry and moderate short stature. Its molecular basis and nosologic status remained unknown. We followed up on those individuals and diagnosed the disorder in an additional member of the family. We used exome sequencing to ascertain the underlying mutation and explored its consequences on three-dimensional models of the affected protein. The MDST phenotype is associated with moderate short stature and knee pain in adults, while extra-skeletal complications are not observed. The sequencing showed that MDST segregated with a c.619T>G single nucleotide transversion in MMP13. The predicted non-conservative amino acid substitution, p.Trp207Gly, disrupts a crucial hydrogen bond in the calcium-binding region of the catalytic domain of the matrix metalloproteinase, MMP13. The MDST phenotype is associated with recessive MMP13 mutations, confirming the importance of this metalloproteinase in the metaphyseal growth plate. Dominant MMP13 mutations have been associated with metaphyseal anadysplasia (OMIM 602111), while a single child homozygous for a MMP13 mutation had been previously diagnosed as "recessive metaphyseal anadysplasia," that we conclude is the same nosologic entity as MDST. Molecular confirmation of MDST allows distinction of it from dominant conditions (e.g., metaphyseal dysplasia, Schmid type; OMIM # 156500) and from more severe multi-system conditions (such as cartilage-hair hypoplasia; OMIM # 250250) and to give precise recurrence risks and prognosis. © 2014 Wiley Periodicals, Inc.

Effect of Nitrogen and Potassium Rates on Early Development of Macaw Palm

ABSTRACT The economic exploitation of macaw palm [Acrocomia aculeate(Jacq.) Lodd. ex Mart.] is currently in transition, from extractivism to agricultural cultivation, thus requiring studies on the fertilization of the crop. This study evaluated the response of three genotypes of macaw palm to increasing rates of nitrogen and potassium, grown in the field until the 2nd year and to establish reference contents of mineral nutrients in the leaf. The experiment was a split-plot randomized block design with five main treatments (N and K rates) and three secondary treatments (genotypes), with three replications, each plot containing three plants. Plant height, leaf number, vigor, and nutrient contents in leaf tissues were evaluated at the end of 2nd year of cultivation. Differential responses were observed among genotypes, indicating that some genotypes are more efficient in the use of mineral inputs. There was a differentiated and positive response to increasing side-dressed N and K rates in the vegetative development of macaw genotypes until the 2nd year of field cultivation, indicating variability in the species in terms of nutrient use efficiency. The N and K fertilization rate corresponding to 360 g N + 480 g K2O per plant, in four split applications over the two years of cultivation, was insufficient to induce maximum vegetative development in the three macaw genotypes. There was no variation in macro- and micronutrient contents in leaf dry matter of the three macaw genotypes.

SWEET SORGHUM PERFORMANCE AFFECTED BY SOIL COMPACTION AND SOWING TIME AS A SECOND CROP IN THE BRAZILIAN CERRADO

ABSTRACT Increasing attention has recently been given to sweet sorghum as a renewable raw material for ethanol production, mainly because its cultivation can be fully mechanized. However, the intensive use of agricultural machinery causes soil structural degradation, especially when performed under inadequate conditions of soil moisture. The aims of this study were to evaluate the physical quality of aLatossolo Vermelho Distroférrico (Oxisol) under compaction and its components on sweet sorghum yield forsecond cropsowing in the Brazilian Cerrado (Brazilian tropical savanna). The experiment was conducted in a randomized block design, in a split plot arrangement, with four replications. Five levels of soil compaction were tested from the passing of a tractor at the following traffic intensities: 0 (absence of additional compaction), 1, 2, 7, and 15 passes over the same spot. The subplots consisted of three different sowing times of sweet sorghum during the off-season of 2013 (20/01, 17/02, and 16/03). Soil physical quality was measured through the least limiting water range (LLWR) and soil water limitation; crop yield and technological parameters were also measured. Monitoring of soil water contents indicated a reduction in the frequency of water content in the soil within the limits of the LLWR (Fwithin) as agricultural traffic increased (T0 = T1 = T2>T7>T15), and crop yield is directly associated with soil water content. The crop sown in January had higher industrial quality; however, there was stalk yield reduction when bulk density was greater than 1.26 Mg m-3, with a maximum yield of 50 Mg ha-1 in this sowing time. Cultivation of sweet sorghum as a second crop is a promising alternative, but care should be taken in cultivation under conditions of pronounced climatic risks, due to low stalk yield.

Water Erosion on an Oxisol under Integrated Crop-Forest Systems in a Transitional Area between the Amazon and Cerrado Biomes

ABSTRACT Water erosion is one of the main factors driving soil degradation, which has large economic and environmental impacts. Agricultural production systems that are able to provide soil and water conservation are of crucial importance in achieving more sustainable use of natural resources, such as soil and water. The aim of this study was to evaluate soil and water losses in different integrated production systems under natural rainfall. Experimental plots under six different land use and cover systems were established in an experimental field of Embrapa Agrossilvipastoril in Sinop, state of Mato Grosso, Brazil, in a Latossolo Vermelho-Amarelo Distrófico (Udox) with clayey texture. The treatments consisted of perennial pasture (PAS), crop-forest integration (CFI), eucalyptus plantation (EUC), soybean and corn crop succession (CRP), no ground cover (NGC), and forest (FRS). Soil losses in the treatments studied were below the soil loss limits (11.1 Mg ha-1 yr-1), with the exception of the plot under bare soil (NGC), which exhibited soil losses 30 % over the tolerance limit. Water losses on NGC, EUC, CRP, PAS, CFI and FRS were 33.8, 2.9, 2.4, 1.7, 2.4, and 0.5 % of the total rainfall during the period of study, respectively.

Ectodysplasin A (EDA)-EDA receptor signalling and its pharmacological modulation

L'ectodysplasine Al (EDA1 ou EDA), un ligand de la famille du TNF, et son récepteur EDAR favorisent le développement des poils, des dents et de plusieurs types de glandes. Chez l'humain, une déficience en EDA cause une dysplasie ectodermique liée à l'X, caractérisée par la genèse défectueuse des phanères. Les souris Tabby, déficientes en Eda, présentent des symptômes similaires. Nous démontrons que les souris Tabby sont en moyenne 7% plus légères que les contrôles au moment du sevrage. Ce phénotype ne dépend pas du génotype des petits, mais exclusivement de celui de la mère, suggérant que l'absence d'EDA perturbe la fonction mammaire. La glande mammaire se développe en plusieurs étapes, principalement à la puberté et pendant la grossesse. Nous avons généré des anticorps pour activer ou inhiber la signalisation d'EDAR. Les anticorps agonistes corrigent le développement de souris ou de chiens déficients en EDA, alors que les antagonistes provoquent une dysplasie ectodermique chez les souris saines. L'exposition répétée de souris Tabby aux anticorps agonistes après le sevrage accroît la taille et la fonction des glandes sébacées, démonstration pharmacologique qu'EDA contrôle l'homéostasie de la glande sébacée adulte. Ces outils seront utiles pour étudier la fonction d'EDA aux diverses étapes du développement de la glande mammaire. Fc-EDAl, un stimulateur d'EDAR, est en phase d'évaluation clinique. Nous avons montré que les structures dépendantes d'EDA qui se forment à différentes étapes du développement répondent à l'action du Fc-EDAl dans des fenêtres temporelles étroites ou larges. De plus, certaines structures peuvent être induites plusieurs jours après le début naturel de leur formation. Alors que la plupart des structures se forment suite à un seul jour d'activation d'EDAR, d'autre demandent un temps de stimulation plus long. La formation des dents est régulée par des signaux activateurs et inhibiteurs. Une forte stimulation d'EDAR spécifiquement appliquée aux deux premières molaires induit des signaux négatifs qui avortent la formation de la troisième molaire, alors qu'une forte stimulation donnée à la troisième molaire la rend hypertrophique tout en induisant parfois une quatrième molaire jamais observée chez les souris de type sauvage ou Tabby. EDA est donc un activateur important de la formation dentaire. Pris dans leur ensemble, ces résultats ont des implications pour la thérapie des dysplasies ectodermiques. - The TNF family ligand Ectodysplasin Al (EDA1 or EDA) and its receptor ED AR regulate embryonic development of hair, teeth and several types of glands. In humans, EDA mutations cause X-linked hypohidrotic ectodermal dysplasia (XLHED), a condition characterized by defective development of skin appendages. £da-deficient (Tabby) mice suffer from similar defects. We observed that Tabby pups at weaning were on average 7% smaller than WT controls, a phenotype that was curiously not linked to the genotype of pups, but to that of mothers, suggesting decreased mammary gland function in the absence of EDA. Mammary glands develop in several steps, most of which are post-natal. We generated monoclonal antibodies to block or activate EDAR signaling. Agonist antibodies rescued developmental defects when administered timely in £cfo-deficient mice and dogs, whereas blocking antibodies induced ectodermal dysplasia in WT mice. Agonist antibodies administered after weaning in £da-deficient mice for several months markedly increased both size and function of sebaceous glands, providing the first demonstration that pharmacological activation of the EDAR pathway in adults can correct important aspects of the dry skin phenotype. This also highlights a role for EDA1 in the homeostasis of adult sebaceous glands. These tools will be useful to study the function of EDA 1 at different stages of mammary gland development. Another EDAR agonist, Fc-EDAl, is currently evaluated in clinical trials. We found that EDA 1-dependent structures forming at different time points during development can respond to Fc-EDAl during time response windows that are narrow or wide. Also, some structures can be triggered up to several days after their normal time of induction. While most structures could be rescued by a single day of EDAR signaling, others required longer exposure times to form. Tooth formation is regulated by activating and inhibitory signals that impact one on the other. When strong EDAR signals were specifically given to the first two molars, overwhelming inhibitory signals completely inhibited formation of the third molar. In contrast, strong signals specifically given to the third molar induced hypertrophy of the later with occasional appearance of a fourth molar never observed in WT or £da-deficient mice. This clearly positions EDA as an important activating signal in tooth formation. Taken together, these results have implications for the therapy of ectodermal dysplasias.

Edar/Eda interactions regulate enamel knot formation in tooth morphogenesis.

tabby and downless mutant mice have apparently identical defects in teeth, hair and sweat glands. Recently, genes responsible for these spontaneous mutations have been identified. downless (Dl) encodes Edar, a novel member of the tumour necrosis factor (TNF) receptor family, containing the characteristic extracellular cysteine rich fold, a single transmembrane region and a death homology domain close to the C terminus. tabby (Ta) encodes ectodysplasin-A (Eda) a type II membrane protein of the TNF ligand family containing an internal collagen-like domain. As predicted by the similarity in adult mutant phenotype and the structure of the proteins, we demonstrate that Eda and Edar specifically interact in vitro. We have compared the expression pattern of Dl and Ta in mouse development, taking the tooth as our model system, and find that they are not expressed in adjacent cells as would have been expected. Teeth develop by a well recorded series of epithelial-mesenchymal interactions, similar to those in hair follicle and sweat gland development, the structures found to be defective in tabby and downless mice. We have analysed the downless mutant teeth in detail, and have traced the defect in cusp morphology back to initial defects in the structure of the tooth enamel knot at E13. Significantly, the defect is distinct from that of the tabby mutant. In the tabby mutant, there is a recognisable but small enamel knot, whereas in the downless mutant the knot is absent, but enamel knot cells are organised into a different shape, the enamel rope, showing altered expression of signalling factors (Shh, Fgf4, Bmp4 and Wnt10b). By adding a soluble form of Edar to tooth germs, we were able to mimic the tabby enamel knot phenotype, demonstrating the involvement of endogenous Eda in tooth development. We could not, however, reproduce the downless phenotype, suggesting the existence of yet another ligand or receptor, or of ligand-independent activation mechanisms for Edar. Changes in the structure of the enamel knot signalling centre in downless tooth germs provide functional data directly linking the enamel knot with tooth cusp morphogenesis. We also show that the Lef1 pathway, thought to be involved in these mutants, functions independently in a parallel pathway.

Biological activity of ectodysplasin A is conditioned by its collagen and heparan sulfate proteoglycan-binding domains.

Mutations in the TNF family ligand EDA1 cause X-linked hypohidrotic ectodermal dysplasia (XLHED), a condition characterized by defective development of skin appendages. The EDA1 protein displays a proteolytic processing site responsible for its conversion to a soluble form, a collagen domain, and a trimeric TNF homology domain (THD) that binds the receptor EDAR. In-frame deletions in the collagen domain reduced the thermal stability of EDA1. Removal of the collagen domain decreased its activity about 100-fold, as measured with natural and engineered EDA1-responsive cell lines. The collagen domain could be functionally replaced by multimerization domains or by cross-linking antibodies, suggesting that it functions as an oligomerization unit. Surprisingly, mature soluble EDA1 containing the collagen domain was poorly active when administered in newborn, EDA-deficient (Tabby) mice. This was due to a short stretch of basic amino acids located at the N terminus of the collagen domain that confers EDA1 with proteoglycan binding ability. In contrast to wild-type EDA1, EDA1 with mutations in this basic sequence was a potent inducer of tail hair development in vivo. Thus, the collagen domain activates EDA1 by multimerization, whereas the proteoglycan-binding domain may restrict the distribution of endogeneous EDA1 in vivo.