寒武纪早期帽状动物壳体Lathamellids类的微细构造及其系统分类

帽状动物壳体 lathamellids 类化石包括 Lathamella caeca, Lathamella sp. nov. 以及 Lathamella symmetrica sp. nov. 三种,目前仅发现于四川峨嵋麦地坪下寒武统麦地坪组上段地层,它们皆以磷质内核方式保存.通过内核化石表面所复制的原始壳体微细构造印痕的研究表明,易漏螺类壳体为双层结构:内层——纤晶层,外层——棱柱层;其壳体原始成分可能为文石质.据上述现象推断,lathamellids 并非为磷质无绞纲腕足类,相反,而与软体动物更为接近,但其在壳腔内具—纵向中突,主要位于壳顶附近,很难与已知的任何一类软体动物直接对比,可能为一类独特的、已经绝灭了的软体动物的1个早期分支.     

Fibrous Aggregates of Short Peptides Containing Two Distinct Aromatic Amino Acid Residues

The aim of the study was the assessment of the ability of short peptides to form aggregates under physiological conditions. The dipeptides studied were derived from different aromatic amino acids (heteroaromatic peptides). Tripeptides were obtained from two distinct aromatic amino acids and cysteine or methionine residue in the C‐terminal, N‐terminal, or central position. The ability of the peptides to form fibrous aggregates under physiological conditions was evaluated using three independent methods: the Congo Red assay, the Thioflavin T assay, and microscopic examinations using normal and polarized light. Materials potentially useful for regenerative medicine were selected based on their cytotoxicity to the endothelial cell line EA.hy 926 and physicochemical properties of films formed by peptides. The required parameters of biocompatibility were fulfilled by H?PheCysTrp?OH, H?PheCysTyr?OH, H?PheTyrMet?OH, and H?TrpTyr?OH.     

Bovine annulus fibrosus hydration affects rate-dependent failure mechanics in tension

The high water content of the intervertebral disc is essential to its load bearing function and viscoelastic mechanical behavior. One of the primary biochemical changes associated with disc degeneration is the loss of proteoglycans, which leads to tissue dehydration. While previous studies have reported the effects of in vivo degeneration on annulus fibrosus (AF) failure mechanics, the independent role of water remains unclear, as does the tissue’s rate-dependent failure response. Our first objective was to determine the effect of loading rate on AF failure properties in tension; our second objective was to quantify the effect of water content on failure properties. Water content was altered through enzymatic digestion of glycosaminoglycans (GAGs) and through osmotic loading. Bovine AF specimens were tested monotonically to failure along the circumferential direction at 0.00697%/s or 6.97%/s. Increased loading rate resulted in a ∼50% increase in linear-region modulus, failure stress, and strain energy density across all treatment groups (p < 0.001). Decreased GAG and water contents resulted in decreased modulus, failure stress, and strain energy density; however, these differences were only observed at the low loading rate (p < 0.05; no changes at high rate). Osmotic loading was used to evaluate the effect of hydration independently from GAG composition, resulting in similar decreases in water content, modulus, and strain energy density. This suggests that hydration is essential for maintaining tissue stiffness and energy absorption capacity, rather than strength, and that GAGs contribute to tissue strength independently from mediating water content.     

Centriolar satellites: molecular characterization, ATP-dependent movement toward centrioles and possible involvement in ciliogenesis

We identified Xenopus pericentriolar material-1 (PCM-1), which had been reported to constitute pericentriolar material, cloned its cDNA, and generated a specific pAb against this molecule. Immunolabeling revealed that PCM-1 was not a pericentriolar material protein, but a specific component of centriolar satellites, morphologically characterized as electron-dense granules, approximately 70-100 nm in diameter, scattered around centrosomes. Using a GFP fusion protein with PCM-1, we found that PCM-1-containing centriolar satellites moved along microtubules toward their minus ends, i.e., toward centrosomes, in live cells, as well as in vitro reconstituted asters. These findings defined centriolar satellites at the molecular level, and explained their pericentriolar localization. Next, to understand the relationship between centriolar satellites and centriolar replication, we examined the expression and subcellular localization of PCM-1 in ciliated epithelial cells during ciliogenesis. When ciliogenesis was induced in mouse nasal respiratory epithelial cells, PCM-1 immunofluorescence was markedly elevated at the apical cytoplasm. At the electron microscopic level, anti-PCM-1 pAb exclusively labeled fibrous granules, but not deuterosomes, both of which have been suggested to play central roles in centriolar replication in ciliogenesis. These findings suggested that centriolar satellites and fibrous granules are identical novel nonmembranous organelles containing PCM-1, which may play some important role(s) in centriolar replication.     

CABYR, a novel calcium-binding tyrosine phosphorylation-regulated fibrous sheath protein involved in capacitation

To reach fertilization competence, sperm undergo an incompletely understood series of morphological and molecular maturational processes, termed capacitation, involving, among other processes, protein tyrosine phosphorylation and increased intracellular calcium. Hyperactivated motility and an ability to undergo the acrosome reaction serve as physiological end points to assess successful capacitation. We report here that acidic (pI 4.0) 86-kDa isoforms of a novel, polymorphic, testis-specific protein, designated calcium-binding tyrosine phosphorylation-regulated protein (CABYR), were tyrosine phosphorylated during in vitro capacitation and bound (45)Ca on 2D gels. Acidic 86-kDa calcium-binding forms of CABYR increased during in vitro capacitation, and calcium binding to these acidic forms was abolished by dephosphorylation with alkaline phosphatase. Six variants of CABYR containing two coding regions (CR-A and CR-B) were cloned from human testis cDNA libraries, including five variants with alternative splice deletions. A motif homologous to the RII dimerization domain of PK-A was present in the N-terminus of CR-A in four CABYR variants. A single putative EF handlike motif was noted in CR-A at aas 197-209, while seven potential tyrosine phosphorylation-like sites were noted in CR-A and four in CR-B. Pro-X-X-Pro (PXXP) modules were identified in the N- and C-termini of CR-A and CR-B. CABYR localizes to the principal piece of the human sperm flagellum in association with the fibrous sheath and is the first demonstration of a sperm protein that gains calcium-binding capacity when phosphorylated during capacitation.     

Modeling alpha-helical coiled-coil interactions: the axial and azimuthal alignment of 1B segments from vimentin intermediate filaments

Attempts at predicting the relative axial alignments of fibrous protein molecules in filamentous structures have relied upon representing the (multichain) molecular structure by a one-dimensional sequence of amino acids. Potential intermolecular ionic and apolar interactions were counted and determined as a function of the relative axial stagger between the molecules. No attempts were made to consider the azimuthal aspect of the interacting molecules and neither were apolar or ionic energy terms used. Surprisingly, this simple approach proved remarkably informative and yielded accurate predictions of the axial periods present. However, a more comprehensive analysis involving the energetics of aggregation taking due regard for the relative azimuths of the molecules as well as their separation should decrease the noise level in the calculations and reveal other pertinent information. Toward that end, we have modeled the interaction between two alpha-helical coiled-coil segments in intermediate filament molecules (1B segments from human vimentin). The relative axial alignment and polarity of the molecules is already known from detailed crosslinking studies and this provides a criterion against which the success (or otherwise) of the modeling can be judged. The results confirm that an antiparallel alignment of two 1B segments is preferred over any of the parallel options (as observed experimentally). The calculated axial alignment, however, is not identical to that observed from detailed crosslinking studies indicating that other parts of the molecule (probably the head and tail domains as well as other coiled-coil segments) have a crucial role in determining the precise mode of axial aggregation. The results also show that the apolar interactions seem to be significantly less important in the alignment process than the ionic ones. This is consistent with the observation of a well-defined period in the linear disposition of the charged (but not apolar) residues along the length of the outer surface of the vimentin molecule.     

The cDNA sequences of three tetrins, the structural proteins of the Tetrahymena oral filaments, show that they are novel cytoskeletal proteins

The oral filaments of the ciliate Tetrahymena consist of the tetrins, insoluble polypeptides with molecular masses of around 85 kD. We characterised the tetrins of T. thermophila by two-dimensional gels and derived a large number of peptide sequences by in gel digestion. Using RT-PCR techniques and RACE-PCR, the complete cDNA sequences of tetrins A, B and C were established. Although tetrins differ strikingly in protein sequence they show a common structural principle. A N-terminal domain of 60 to 100 residues contains most of the proline residues of the tetrins and is probably globular. It is followed by a long alpha-helical domain of 620 to 640 residues which either lacks prolines or in tetrin A contains a single proline residue. Although this long domain has coiled coil forming ability, the individual heptad repeats are not extensive. Tetrins are novel cytoskeletal proteins unique to ciliates. Since the three tetrin sequences account for all 900 amino acid residues obtained by microsequencing of peptides, an additional major tetrin seems excluded. A minor component D is related to tetrin B by peptide sequences. The isoelectric variants, particularly obvious for tetrin A, most likely reflect post-translational modifications. These could arise by phosphorylation of serines and threonines in the proline rich N-terminal domain.     

Rodlet cells in epidermal explant cultures of Lepomis Macrochirus

Sunfish rodlet cells were examined in vitro using a novel tissue explant system. Outgrowth of epidermal cell layers from explanted fish scales enabled both live cell videomicroscopy and immunocytochemical analysis of rodlet cells within the cell layer. Cells stained with fluorescent phallotoxin and antibody to tubulin showed that F‐actin is a component of the fibrous capsule that envelopes the cell and a microtubule network extends from the basal to apical ends of the cell interior. The fibrous capsule is also enriched for phosphotyrosine suggesting a potential signal‐transducing capability is present in this structure. Videomicroscopy analysis of live explant cultures demonstrated that rodlet cells are immobile and that interior structures are highly dynamic. Rodlet sacs can undergo extension and retraction, while intracellular particles can move rapidly within these cells. Fish scale tissue explants provide a useful system for analyzing the molecular composition and dynamic behavior of rodlet cells.