Fibulin-5/DANCE has an elastogenic organizer activity that is abrogated by proteolytic cleavage in vivo

Elastic fibers are required for the elasticity and integrity of various organs. We and others previously showed that fibulin-5 (also called developing arteries and neural crest EGF-like [DANCE] or embryonic vascular EGF-like repeat–containing protein [EVEC]) is indispensable for elastogenesis by studying fibulin-5–deficient mice, which recapitulate human aging phenotypes caused by disorganized elastic fibers (Nakamura, T., P.R. Lozano, Y. Ikeda, Y. Iwanaga, A. Hinek, S. Minamisawa, C.F. Cheng, K. Kobuke, N. Dalton, Y. Takada, et al. 2002. Nature. 415:171–175; Yanagisawa, H., E.C. Davis, B.C. Starcher, T. Ouchi, M. Yanagisawa, J.A. Richardson, and E.N. Olson. 2002. Nature. 415:168–171). However, the molecular mechanism by which fiblin-5 contributes to elastogenesis remains unknown. We report that fibulin-5 protein potently induces elastic fiber assembly and maturation by organizing tropoelastin and cross-linking enzymes onto microfibrils. Deposition of fibulin-5 on microfibrils promotes coacervation and alignment of tropoelastins on microfibrils, and also facilitates cross-linking of tropoelastin by tethering lysyl oxidase-like 1, 2, and 4 enzymes. Notably, recombinant fibulin-5 protein induced elastogenesis even in serum-free conditions, although elastogenesis in cell culture has been believed to be serum-dependent. Moreover, the amount of full-length fibulin-5 diminishes with age, while truncated fibulin-5, which cannot promote elastogenesis, increases. These data suggest that fibulin-5 could be a novel therapeutic target for elastic fiber regeneration

Distinct temporal integration of noradrenaline signaling by astrocytic second messengers during vigilance

Astrocytes may function as mediators of the impact of noradrenaline on neuronal function. Activation of glial α1-adrenergic receptors triggers rapid astrocytic Ca2+ elevation and facilitates synaptic plasticity, while activation of β-adrenergic receptors elevates cAMP levels and modulates memory consolidation. However, the dynamics of these processes in behaving mice remain unexplored, as do the interactions between the distinct second messenger pathways. Here we simultaneously monitored astrocytic Ca2+ and cAMP and demonstrate that astrocytic second messengers are regulated in a temporally distinct manner. In behaving mice, we found that while an abrupt facial air puff triggered transient increases in noradrenaline release and large cytosolic astrocytic Ca2+ elevations, cAMP changes were not detectable. By contrast, repeated aversive stimuli that lead to prolonged periods of vigilance were accompanied by robust noradrenergic axonal activity and gradual sustained cAMP increases. Our findings suggest distinct astrocytic signaling pathways can integrate noradrenergic activity during vigilance states to mediate distinct functions supporting memory

A Comprehensive Resource of Interacting Protein Regions for Refining Human Transcription Factor Networks

Large-scale data sets of protein-protein interactions (PPIs) are a valuable resource for mapping and analysis of the topological and dynamic features of interactome networks. The currently available large-scale PPI data sets only contain information on interaction partners. The data presented in this study also include the sequences involved in the interactions (i.e., the interacting regions, IRs) suggested to correspond to functional and structural domains. Here we present the first large-scale IR data set obtained using mRNA display for 50 human transcription factors (TFs), including 12 transcription-related proteins. The core data set (966 IRs; 943 PPIs) displays a verification rate of 70%. Analysis of the IR data set revealed the existence of IRs that interact with multiple partners. Furthermore, these IRs were preferentially associated with intrinsic disorder. This finding supports the hypothesis that intrinsically disordered regions play a major role in the dynamics and diversity of TF networks through their ability to structurally adapt to and bind with multiple partners. Accordingly, this domain-based interaction resource represents an important step in refining protein interactions and networks at the domain level and in associating network analysis with biological structure and function

NUTRITIONAL REQUIREMENTS OF CORYNEBACTERIUM RENALE

The growth factor and amino acid requirements of Corynebacterium renale were investigated with special reference to the 3 types of this species reported by YANAGAWA et al. (1967). The results are summarized as follows. 1) Strains of type I required thiamine, biotin and pantothenic acid for maximum growth. These vitamins acted as growth stimulatory factors. Strains of type II required biotin, nicotinic acid and p-aminobenzoic acid as essential growth factors. Thiamine and pantothenic acid acted as growth stimulants. Strains of type III which was most exacting, required thiamine, biotin, nicotinic acid, pantothenic acid and pyridoxine as essential growth factors. Without even one of these vitamins type III strains did not grow. Pyridoxine was required by type III but not by type II while p-aminobenzoic acid was essential for type II but not for type III. 2) Sodium bicarbonate stimulated the growth of all 3 types of C. renale. 3) Ammonium sulfate was necessary for maximum growth of each type. 4) The essential amino acid common to the 3 types of C. renale included glutamic acid, valine and isoleucine, Type I required only these 3 amino acids. Type II required the 3 amino acids essentially ; the growth was accelerated with the addition of methionine, phenylalanine and histidine. Growth of type III was most exacting. In addition to the common 3 amino acids, it required tryptophane essentially. Methionine, phenylalanine and arginine were also needed for maximum growth. Thus, in complexity, the requirements of both growth factors and amino acids was as follows : types III, II and I. 5) Growth inhibition by cystine was particularly noticeable in type I. 6) There was discussion on the comparison of the nutritional requirements of C. diphtheriae, C. sepedonicum and C. renale

A CINEMATOGRAPHIC STUDY OF THE PENETRATION OF CULTURED CELLS BY TOXOPLASMA GONDII

Cinematographic observation with phase contrast microscopy was undertaken to elucidate a course of penetration of cultured cells by Toxoplasma gondii (RH strain). The results are summarized as follows : 1) The parasites which moved toward the cells showed active, irregular movements. No penetration was observed by less active or completely inactived parasites. 2) In the course of penetration of the cell membrane, the parasite attached itself to the cell membrane at a pole of its major axis. The attached portion of the parasite formed a rostrum with which it seemed to bore a small hole in the cell membrane through which it entered the cytoplasm. The hole or invagination must be smaller than the parasite because when passing through, the parasite was constricted. The time required for penetration of the cell averaged about 40 seconds. 3) Following the penetration, the parasites moved slowly in direction of the cell nucleus, but none were observed to penetrate the nuclei. 4) No common, definite cellular response was observed but, in general, the movement of the cytoplasmic granules surrounding the parasites decreased with time. 5) Escape of toxoplasma from the infected cell was observed, which was due to the movement of toxoplasma itself. Since the cell from which the toxoplasma escaped was found rather inactive, it is probable toxoplasma is highly sensitive to the nutritional condition of host cell

HISTOPATHOLOGICAL SURVEY OF PROTOZOA, HELMINTHS AND ACARIDS OF IMPORTED AND LOCAL PSITTACINE AND PASSERINE BIRDS IN JAPAN

A total of 534 psittacine and passerine birds consisting of 241 imported and 293 local birds were examined histologically. As a result, the following parasites were found : Giardia (86 cases), Knemido-coptes (26 cases), coccidia (10 cases), Ascaridia (6 cases), Cryptosporidium (5 cases), Sarcocystis (5 cases), tapeworm (4 cases), microfilaria (2 cases), Hexamita (1 case), and Spiroptera (1 case). High incidences of giardiasis and knemido-coptic infestation were detected in the local birds, but rarely in the imported birds. Giardial trophozoites were observed mainly in the duodenum of budgerigars (Melopsittacus undulatus). Knemidocoptic mites burrowed into the epidermis producing proliferative dermatitis in 25 budgerigars and l African Grey Parrot (Psittacus erithacus erithacus). This ectoparasite often infested the skin around the cloaca. Coccidiosis was seen only in the small intestines of the finch (Poephila gouldiae gouldiae), African Grey Parrot, Rainbow lory (Trichoglossus haematodus), Indian Ring-necked parakeet (Psittacula krameri manillensis) and peach-faced lovebird (Agapornis roseicollis). Two parrots (Amazona aestiva aestiva and Psittacus erithacus erithacus) and two budgerigars had intestinal cryptosporidiosis. Conjunctivitis associated with cryptosporidial infection was seen in a lovebird. Sarcocystis cysts containing crescent-shaped bradyzoites were found not only in the thigh and breast but also in the heart and cloacal muscles. Other organisms such as Ascaridia, tapeworm, microfilaria, Hexamita, and Spiroptera were clinically less significant. However, infections such as Giardia and Cryptosporidim might have zoonotic implications