The vascular-associated lymphoid tissue: a new site of local immunity

Recent data suggest that atherosclerosis might be a systemic (auto)immune reaction against heat shock protein 60, first occurring at notorious local predilection sites, i.e. the intima at arterial branching points. The local infiltration of mononuclear cells, mainly macrophage-derived foam cells, T cells and smooth-muscle cells in atheromatous plaques, have long been described. During the past few years, research has been concentrated on the early stages in the development of atherosclerosis, and on healthy arteries from young individuals unaffected by arterial disease. In this review, we summarize data characterizing pre-existing mononuclear cell infiltrations in healthy arteries from children and teenagers. These arterial accumulations at regions known to be predilection sites for the later development of atherosclerosis consist mostly of activated T cells, macrophages and dendritic cells, with only a few mast cells and virtually no B or natural killer cells. In analogy to the mucosa-associated lymphoid tissue, we termed these accumulations 'vascular-associated lymphoid tissue', and assumed a similar function as a local immunosurveillance system, monitoring the bloodstream for potentially harmful endogenous or exogenous antigens. In addition to the remarkable accumulation of mononuclear cells, the vascular-associated lymphoid tissue regions are characterized by a typical distribution of extracellular matrix proteins: collagen type I, collagen type III, fibronectin and tenascin are expressed preferentially in the vascular-associated lymphoid tissue region, whereas collagen type IV, collagen type V, collagen type VI and laminin show a homogenous distribution throughout all regions of the intima. Vascular adhesion molecules type 1, intercellular adhesion molecules type 1 and P-selectin are already present on the healthy endothelial cells of young children. Interactions between adhesion molecules, extracellular matrix components and cellular elements of the vascular-associated lymphoid tissue may provide the basis for the cellular accumulations in the vascular-associated lymphoid tissue regions and the possible development of atherosclerotic lesions later in life.     

Optimality principles in arterial branching

A comparative study of four optimality principles for the branching geometry of blood arteries is presented. The results offer four different criteria which can be tested by experimental data to establish which of these principles is followed in the cardiovascular system. More significantly, the results suggest the further possibility that the geometry of arterial junctions may be governed by all of these principles simultaneously, to thus achieve a much higher degree of optimality than has hitherto been suspected. This result offers a basis for seeking a correlation between the degree of optimality of a particular junction and the incidence of certain arterial lesions at that junction.     

The subscapular arterial tree as a source of microvascular arterial grafts

The subscapular arterial tree may be used as a source of microvascular grafts to replace damaged or diseased portions of arteries, particularly in the hand and forearm. By studying cadaver dissections, it is possible to estimate the number of branches that may be found at different arterial segment lengths from the origin of the subscapular artery. Fifty-five preserved cadaver subscapular arterial trees were dissected, and the branching patterns were documented. Three major arterial branching patterns of the subscapular artery were observed with one, two, and three major branches to the serratus anterior in 60 percent, 29 percent, and 9 percent of the cases, respectively. The authors determined the number of 1-mm-diameter, 1-cm-long branches arising from each of six 3-cm regions of the arterial tree measured from the origin of the subscapular artery to the end of the longest terminal branch. The probability of finding at least one usable terminal branch that is at least 12.0 cm in length was found to be 98 percent. Typically, there are two to five useful branches at this distance. Such information may help surgeons fine tune their process of selecting an appropriate arterial donor site for a particular arterial defect and supports the use of the subscapular arterial tree as a donor site for microvascular arterial grafts.     

Arterial branching within the confines of fractal L-system formalism

Parametric Lindenmayer systems (L-systems) are formulated to generate branching tree structures that can incorporate the physiological laws of arterial branching. By construction, the generated trees are de facto fractal structures, and with appropriate choice of parameters, they can be made to exhibit some of the branching patterns of arterial trees, particularly those with a preponderant value of the asymmetry ratio. The question of whether arterial trees in general have these fractal characteristics is examined by comparison of pattern with vasculature from the cardiovascular system. The results suggest that parametric L-systems can be used to produce fractal tree structures but not with the variability in branching parameters observed in arterial trees. These parameters include the asymmetry ratio, the area ratio, branch diameters, and branching angles. The key issue is that the source of variability in these parameters is not known and, hence, it cannot be accurately reproduced in a model. L-systems with a random choice of parameters can be made to mimic some of the observed variability, but the legitimacy of that choice is not clear.     

Ephrin-As play a rhombomere-specific role in trigeminal motor axon projections in the chick embryo

In this study, we investigate the possible role of ephrin-Eph signaling in trigeminal motor axon projections. We find that EphA receptors are expressed at higher levels by rhombomere 2 (r2) trigeminal motor neurons than by r3 trigeminal motor neurons in the chick embryo. Mapping of rhombomere-specific axon projections shows that r2 and r3 trigeminal motor neurons project to different muscle targets, including the mandibular adductor and the intermandibularis muscles respectively. Ephrin-A5 is expressed in these muscles, especially in some regions of the intermandibularis muscle, and can cause growth cone collapse of both r2 and r3 motor axons in vitro. We demonstrate that in vivo overexpression of ephrin-A5 in the intermandibularis muscle, or overexpression of dominant-negative EphA receptors in trigeminal motor neurons leads to a reduction in branching of r3-derived motor axons specifically. Overexpression of full-length EphA receptors impairs the formation of r3 projections to the intermandibularis muscle. These findings indicate that ephrins and their Eph receptors play a role in trigeminal motor axon topographic mapping and in rhombomere 3-derived projections in particular.     

Deciphering interfungal relationships in the 410 million-year-old Rhynie chert: Glomoid spores under attack

Various types of other fungi colonize glomeromycotinan (Mucoromycota) spores in the Early Devonian Rhynie chert. However, relatively few of these associations have been described and evaluated in detail. One particular type of glomoid spore located in degrading land plant axes from the Rhynie chert provides evidence of (simultaneous) interaction with three different fungi. Massive callosities occur around the intrusion filaments of a chytrid-like parasite with epibiotic sporangia, while the hyphae of a delicate mycelial fungus extend into the spore lumen without triggering a recognizable host response. Several spores show large numbers of inwardly directed projections, which are regularly distributed and consist of a short hyphal branch encased in host wall material. The projections represent the penetration sites of a distinctive, mycelial fungus that forms a mantle-like hyphal sheath around the spores. This type of fungal interaction with glomeromycotinan spores has not previously been reported, and thus expands our knowledge of the numerous interfungal relationships that existed in early continental ecosystems.     

Dominance of geometric overelastic factors in pulse transmission through arterial branching

The branching characteristic of the arterial system is such that blood pressure pulses propagate with minimum loss. This characteristic depends on the geometric and elastic properties of branching vessels. In the current investigation, mathematical relations of branching geometry and elastic properties are formulated and their relative contributions to pulse reflection at an arterial junction are analyzed. Results show that alteration of pulse transmission through the junction is more significantly affected by changes in branching vessel radii and wall thickness than by corresponding percentage changes in vessel wall elastic moduli.     

A perfusion-independent role of blood vessels in determining branching stereotypy of lung airways

Blood vessels have been shown to play perfusion-independent roles in organogenesis. Here, we examined whether blood vessels determine branching stereotypy of the mouse lung airways in which coordinated branching of epithelial and vascular tubes culminates in their co-alignment. Using different ablative strategies to eliminate the lung vasculature, both in vivo and in lung explants, we show that proximity to the vasculature is indeed essential for patterning airway branching. Remarkably, although epithelial branching per se proceeded at a nearly normal rate, branching stereotypy was dramatically perturbed following vascular ablation. Specifically, branching events requiring a rotation to change the branching plane were selectively affected. This was evidenced by either the complete absence or the shallow angle of their projections, with both events contributing to an overall flat lung morphology. Vascular ablation also led to a high frequency of ectopic branching. Regain of vascularization fully rescued arrested airway branching and restored normal lung size and its three-dimensional architecture. This role of the vasculature is independent of perfusion, flow or blood-borne substances. Inhibition of normal branching resulting from vascular loss could be explained in part by perturbing the unique spatial expression pattern of the key branching mediator FGF10 and by misregulated expression of the branching regulators Shh and sprouty2. Together, these findings uncovered a novel role of the vasculature in organogenesis, namely, determining stereotypy of epithelial branching morphogenesis.     

Input impedance for studying hydraulic parameters of the vessel system

Vascular input impedance can be used as an effective tool in estimating hydraulic parameters of arterial bed. These parameters may be interpreted as hydraulic resistance, elastance and inertance of particular sites of the arterial system. There is no significant difference between these parameters and those obtained through a direct measurement.     

Neuropilar Projections of the Anterior Gastric Receptor Neuron in the Stomatogastric Ganglion of the Jonah Crab,Cancer Borealis

Sensory neurons provide important feedback to pattern-generating motor systems. In the crustacean stomatogastric nervous system (STNS), feedback from the anterior gastric receptor (AGR), a muscle receptor neuron, shapes the activity of motor circuits in the stomatogastric ganglion (STG) via polysynaptic pathways involving anterior ganglia. The AGR soma is located in the dorsal ventricular nerve posterior to the STG and it has been thought that its axon passes through the STG without making contacts. Using high-resolution confocal microscopy with dye-filled neurons, we show here that AGR from the crab Cancer borealis also has local projections within the STG and that these projections form candidate contact sites with STG motor neurons or with descending input fibers from other ganglia. We develop and exploit a new masking method that allows us to potentially separate presynaptic and postsynaptic staining of synaptic markers. The AGR processes in the STG show diversity in shape, number of branches and branching structure. The number of AGR projections in the STG ranges from one to three simple to multiply branched processes. The projections come in close contact with gastric motor neurons and descending neurons and may also be electrically coupled to other neurons of the STNS. Thus, in addition to well described long-loop pathways, it is possible that AGR is involved in integration and pattern regulation directly in the STG.     

Arguments justifying the acception of the notion "sublobular artery" in the nomenclature of renal vessels: the "conical" architectonics of the renal cortex

The corrosion technique was used to demonstrate the presence of the "sublobular artery", a constant vessel with a special profile of the dog kidney, generating after two divisions the so-called interlobular arteries as a third generation of branches. More rarely arteriae interlobulares emerge also directly from the arcuate artery. The sublobular arteries (= parent vessels of the interlobular arteries) are characterized by their particular origin, branching type, and by having no or sporadically disposed glomeruli. For this reason, the authors suggest the acception of the term "arteria sublobularis" in the international nomenclature of renal vessels. According to authors' opinion the dog kidney is composed of a number of conical vascular-parenchymatous units, each containing in their axis a sublobular artery of the second order ("conical" architectonics of the renal cortex).     

Oligosaccharide microheterogeneity of the murine major histocompatibility antigens. Reproducible site-specific patterns of sialylation and branching in asparagine-linked oligosaccharides

The influence of peptide structure of endogenous cell-surface glycoproteins on the branching and sialylation of their asparagine-linked oligosaccharides was evaluated in a murine B cell lymphoma, AKTB-1b. This cell line simultaneously synthesizes two classes of major histocompatibility antigens that, within each class, share a high degree of amino acid sequence homology and possess potential N-linked glycosylation sites at invariant positions. [3H]Mannose-labeled oligosaccharides were released from each of 11 purified glycosylation sites by the almond peptide:N-glycosidase and analyzed by a variety of chromatographic procedures and glycosidase treatments. The data indicate: 1) a unique distribution of oligosaccharide structures is present at each glycosylation site; 2) each site-specific oligosaccharide pattern is highly reproducible, independent of the number of in vivo tumor passages. The heavy chain of the class I antigens, H-2Kk and H-2Dk contain two and three sites, respectively, in which biantennary structures predominate. However, each site varies with respect to the extent of sialylation and the proportions of more highly branched structures present. The class II antigens, I-Ak and I-Ek, each contain an alpha-chain site toward the N terminus and a single beta-chain site where the overall extent of sialylation is similar, yet the distributions of antennary structures are dramatically different for each. The alpha-chains of each class II antigen also contain a more C-terminal underglycosylated site where sialylation and branching are reduced to differing degrees depending upon the site. The influence of peptide structure on oligosaccharide microheterogeneity is manifest at two levels. First, the overall distributions of oligosaccharides at corresponding sites on structurally related glycoproteins are similar. Second, the specific "fingerprint" of sialylation and branching patterns at a particular site are reproducibly unique. These data suggest that subtle changes in peptide structure are reflected in the extent of sialylation and branching of oligosaccharides found at corresponding glycosylation sites of structurally related glycoproteins.     

Similarities and gradients in growth unit branching patterns during ontogeny in "Fuji" apple trees: a stochastic approach

This study aims to explore and model the changes in growth unit (GU) branching patterns during tree ontogeny. The question was addressed in apple trees cv. "Fuji", by analysing the relative impact of GU length and within-tree position. The development of two 6-year-old trees was recorded over 6 years. The fate of axillary buds along each GU was represented as a sequence of symbols corresponding to five types of lateral growth: latent buds, short, medium, long, and floral lateral GUs. Based on an exploratory analysis of data and a priori hypotheses, a hidden semi-Markov chain was estimated from all of these GU sequences. This model was composed of six transient states representing successive branching zones along the GUs. The accuracy of this global model was a posteriori assessed by fitting the characteristic distributions computed from model parameters to the corresponding empirical characteristic distributions extracted from the observed sequences. The observed sequences were then grouped hierarchically according to the GU length, year of growth, and branching order. Comparing model parameters between these sub-groups revealed similarities between GUs. These similarities were based on particular branching zones whose composition and relative position within the GUs remained invariant across the subgroups: the latent zones, floral zone, and short-lateral zone. The probability of occurrence of the floral zone varied with the year, showing the alternate fruiting of "Fuji". It is shown that, during tree ontogeny, as GU length decreases, branching patterns tend to progressively simplify due to the disappearance of the most central zones and a progressive reduction in the length of the floral zone.     

Synapse‐dependent and independent mechanisms of thalamocortical axon branching are regulated by neuronal activity

Axon branching and synapse formation are critical processes for establishing precise circuit connectivity. These processes are tightly regulated by neural activity, but the relationship between them remains largely unclear. We use organotypic coculture preparations to examine the role of synapse formation in the activity‐dependent axon branching of thalamocortical (TC) projections. To visualize TC axons and their presynaptic sites, two plasmids encoding DsRed and EGFP‐tagged synaptophysin (SYP‐EGFP) were cotransfected into a small number of thalamic neurons. Time‐lapse imaging of individual TC axons showed that most branches emerged from SYP‐EGFP puncta, indicating that synapse formation precedes emergences of axonal branches. We also investigated the effects of neuronal activity on axon branching and synapse formation by manipulating spontaneous firing activity of thalamic cells. An inward rectifying potassium channel, Kir2.1, and a bacterial voltage‐gated sodium channel, NaChBac, were used to suppress and promote firing activity, respectively. We found suppressing neural activity reduced both axon branching and synapse formation. In contrast, increasing neural activity promoted only axonal branch formation. Time‐lapse imaging of NaChBac‐expressing cells further revealed that new branches frequently appeared from the locations other than SYP‐EGFP puncta, indicating that enhancing activity promotes axonal branch formation due to an increase of branch emergence at nonsynaptic sites. These results suggest that presynaptic locations are hotspots for branch emergence, and that frequent firing activity can shift branch emergence to a synapse‐independent process. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 323–336, 2016     

Modeling of intraorgan arterial vasculatures. II. Propagation of pressure waves

The dependences of the wave conductivity of self-similar dichotomous branching models of intraorgan arterial vasculatures on the model parameters have been calculated. It was found that, with different sets of parameters, it is possible to model the sucking effect connected with negative wave reflection at the arterial branching as well as the resonant properties of arterial beds. It was shown that the selection of an adequate model for a given intraorgan vasculature should be based on the agreement between the biophysical characteristics of the model and the vasculature that reflect the propagation and reflection of pulse waves.     

Cost analysis of arterial branching in the cardiovascular systems of man and animals

A new scheme is presented whereby data on arterial branching can be interpreted in terms of direct cost to the physiological system. The scheme makes it possible to assess, at a glance, the true degree of optimality of an arterial network. Departure from optimality is indicated in terms of cost, rather than in terms of the difference between theoretical and measured branching angles. The scheme is applied to several groups of biological data and new conclusions are reached with regard to their degrees of optimality.     

Combined action of Escherichia coli glycogen synthase and branching enzyme in the so-called "unprimed" polyglucoside synthesis.

Mutants of Escherichia coli which are unable to synthesize glycogen were used to study the so-called “unprimed” synthesis of glycogen. The glycogen synthase has been partially purified from these mutants. During the purification, attempts were made to separate the activity which requires the addition of an exogenous primer (primed activity) from the activity which does not require a primer but is highly dependent on the presence of some salts such as citrate and EDTA (unprimed activity). No separation between these two activities could be achieved but the results obtained by chromatography on DEAE-Sephadex indicate that there is a single form of glycogen synthase which is responsible for both unprimed and primed activity. The evidence that a single protein was necessary to catalyze these two reactions was given by the findings that mutants defective in glycogen synthase activity were unable to catalyze glucosyl transfer without added primer. At low concentration, the glycogen synthase purified from a branching enzyme negative mutant catalyzed the unprimed reaction at a slow rate even in presence of salts. A protein activator of this reaction was found in mutants lacking glycogen synthase but not in mutants lacking branching enzyme. The hypothesis that this activator is the branching enzyme itself was supported by the observation that it co-purified with the branching enzyme from a E. coli strain defective in glycogen synthase activity. EDTA or Triton X-100 increased the stimulation of the unprimed synthesis by the branching enzyme. The apparent affinity of the glycogen synthase for glycogen was increased twofold in the presence of EDTA but the branching enzyme further increased the effect of EDTA. The combined action of the glycogen synthase and the branching enzyme on the endogenous glucan associated with the synthase may account for the unprimed activity observed in vitro.     

Cost of departure from optimality in arterial branching

Measurements of branching angles in the arterial tree have in the past indicated a great deal of scatter away from what is expected to be optimum on theoretical grounds. In this study the cost penalty of nonoptimum branching angles is calculated for the first time to determine how far from optimum these angles are. The results lead to the remarkable conclusion that while the scatter of the measured branching angles is fairly large, they represent deviations from optimum angles which correspond to only 2% or so penalty in cost.     

Neurite consolidation is an active process requiring constant repression of protrusive activity

During development, neurons extend projections that pathfind to reach their appropriate targets. These projections are composed of two distinct domains: a highly dynamic growth cone and a stable neurite shaft, which is considered to be consolidated. Although the regulation of these domains is critical to the appropriate formation of neural networks, the molecular mechanisms that regulate neurite shape remain poorly understood. Here, we show that calpain protease activity localizes to the neurite shaft, where it is essential for the repression of protrusive activity by limiting cortactin levels and inhibiting actin polymerization. Correspondingly, inhibition of calpain by branching factors induces the formation of new growth cones along the neurite shaft through cAMP elevation. These findings demonstrate that neurite consolidation is an active process requiring constant repression of protrusive activity. We also show that sprouting is, at least in part, accomplished by turning off the mechanism of consolidation.