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51.
A low-resolution study of testicular lactate dehydrogenase using the molecular replacement technique
W. Donald L. Musick April D. Adams Michael G. Rossmann Thomas E. Wheat Erwin Goldberg 《Journal of molecular biology》1976,104(3):659-668
The orientation of the molecular 2-fold axes of mouse testicular lactate dehydrogenase (LDHase3-C4) was determined by a rotation function search. These were subsequently identified with the P, Q, and R axes of dogfish LDHase-M4. Since LDHase-C4 crystallized with one molecule in a triclinic cell, the origin of the co-ordinate system was arbitrarily fixed at the molecular center. Structure factor phases were derived from an appropriately oriented dogfish apo LDHase-M4 phasing model and combined with the observed structure amplitudes to produce a hybrid electron density map. Density points related by the molecular 222 point symmetry were averaged so as to remove the bias of the phasing model. At 7.5 Å resolution, the structure of the crystallized mouse LDHase-C4 was found to be without coenzyme, with a conformation indistinguishable from that of dogfish apo LDHase-M4. 相似文献
52.
Bob?JA?Schijvenaars Barend?Mons Marc?Weeber Martijn?J?Schuemie Erik?M?van Mulligen Hester?M?Wain Jan?A?KorsEmail author 《BMC bioinformatics》2005,6(1):149
Background
Massive text mining of the biological literature holds great promise of relating disparate information and discovering new knowledge. However, disambiguation of gene symbols is a major bottleneck. 相似文献53.
Miguel L Soares Seiki Haraguchi Maria-Elena Torres-Padilla Tibor Kalmar Lee Carpenter Graham Bell Alastair Morrison Christopher JA Ring Neil J Clarke David M Glover Magdalena Zernicka-Goetz 《BMC developmental biology》2005,5(1):1-11
Background
Studies of gene function in the mouse have relied mainly on gene targeting via homologous recombination. However, this approach is difficult to apply in specific windows of time, and to simultaneously knock-down multiple genes. Here we report an efficient method for dsRNA-mediated gene silencing in late cleavage-stage mouse embryos that permits examination of phenotypes at post-implantation stages.Results
We show that introduction of Bmp4 dsRNA into intact blastocysts by electroporation recapitulates the genetic Bmp4 null phenotype at gastrulation. It also reveals a novel role for Bmp4 in the regulation the anterior visceral endoderm specific gene expression and its positioning. We also show that RNAi can be used to simultaneously target several genes. When applied to the three murine isoforms of Dishevelled, it leads to earlier defects than previously observed in double knock-outs. These include severe delays in post-implantation development and defects in the anterior midline and neural folds at headfold stages.Conclusion
Our results indicate that the BMP4 signalling pathway contributes to the development of the anterior visceral endoderm, and reveal an early functional redundancy between the products of the murine Dishevelled genes. The proposed approach constitutes a powerful tool to screen the functions of genes that govern the development of the mouse embryo. 相似文献54.
Specimens of the mesoparasitic barnacle, Anelasma, were found attached to several host deep-sea squaloid sharks, Centroscyllium fabricii from off Greenland, Etmopterus princeps from the Canary Islands, and Etmopterus unicolor and Etmopterus granulosus from Three Kings Ridge and Louisville Ridge near New Zealand. One to three parasites were partially embedded in each host shark in the head, mouth, bases of 1st and 2nd dorsal fins, pectoral and pelvic fins, caudal fin, abdomen or claspers. Testes and claspers of parasitized mature-sized males were less developed than those of non-parasitized mature males. The ova of parasitized females of mature size were small and whitish-yellow compared to the large yellow ova of non-parasitized mature females. The number of mature ova in a parasitized specimen was lower than expected in relation to a non-parasitized individual of the same size. Attachment of parasitic barnacles appeared to retard the development of reproductive organs of host sharks. 相似文献
55.
We have reprogrammed the stimulus-responsive conformational change property of a virus nanoparticle (VNP) to enable the surface exposure of metal binding motifs upon activation with heat. The VNP is based on the widely investigated adeno-associated virus (AAV). An intrinsic bioactive functionality of AAV was genetically replaced with a hexahistidine (His) tag. The peptide domain with the inserted His tag is normally inaccessible. Upon external stimulation with heat, the VNP undergoes a conformational change, resulting in externalization of His tag-containing domains and the conferred ability to bind metal. We show that beyond this newfound functionality of the capsid, the VNPs maintain many of the wild-type capsid properties. Our work lays the groundwork for developing stimulus-responsive VNPs that can be used as "smart" building blocks for the creation of higher order structures. 相似文献
56.
In search for an alternative for nutritional antimicrobials in piglet feeding, the effects of adding whole Cuphea seeds, as a natural source of medium chain fatty acids (MCFA), with known antimicrobial effects, and an exogenous lipase to a weaner diet were studied. The foregut flora, the gut morphology, some digestive parameters and the zootechnical performance of weaned piglets were investigated. Thirty newly weaned piglets, initial weight 7.0 ± 0.4 kg, were divided according to litter, sex and weight in two groups (control diet; Cuphea+lipase diet). The Cuphea seeds (lanceolata and ignea) (50 g kg?1) were substituted for soybean oil (15 g kg?1), Alphacell (25 g kg?1) and soy protein isolate (10 g kg?1) in the control diet. Also 500 mg kg?1 microbial lipase was added to the Cuphea diet. The piglets were weighted individually on days 0, 3, 7, 14 and 16. Feed intake was recorded per pen during days 0 to 3, 3 to 7, 7 to 14 and 14 to 16. On day 7 five piglets of each experimental group were euthanized for counting the gastric and small intestinal gut flora and for gut morphology at two sites of the small intestine (proximal, distal). The results indicate a trend towards improved performances parameters by feeding Cuphea + lipase. The enzymic released MCFA (1.7 g kg?1 fresh gastric contents) tended to decrease the number of Coliforms in the proximal small intestine, but increased the number in the stomach and distal small intestine. With Cuphea, the number of Streptococci was significantly lower in small intestine, but not in the stomach, while the number of Lactobacilli was significantly lower in the distal small intestine and tended to be lower in the stomach and proximal small intestine. No differences between the diets were noted for the total anaerobic microbial load in the stomach or in the gut. Feeding Cuphea+lipase resulted in a significantly greater villus height (distal small intestine) and a lesser crypt depth (proximal and distal small intestine) and greater villus/crypt ratio depth (proximal and distal small intestine). The intra-epithelial lymphocyte (IEL) counts per 100 enterocytes were significantly decreased in the proximal small intestine and tended to decrease in the distal small intestine by feeding the Cuphea+lipase diet. Both phenomena are indicative for a more healthy and better functional state of the mucosa. Present results are in line with foregoing research, showing that manipulation of the gut ecosystem by the enzymic in situ released MCFA in the stomach and foregut can result in improved performances of the piglets, which makes the concept a potential alternative for in-feed nutritional antibiotics. 相似文献
57.
The resting metabolic rates (R(R)) of western North Atlantic Ocean sciaenids, such as Atlantic croaker Micropogonias undulatus, spot Leiostomus xanthurus and kingfishes Menticirrhus spp., as well as the active metabolic rates (R(A)) of M. undulatus and L. xanthurus were investigated to facilitate inter and intraspecific comparisons of their energetic ecology. The R(R) of M. undulatus and L. xanthurus were typical for fishes with similar lifestyles. The R(R) of Menticirrhus spp. were elevated relative to those of M. undulatus and L. xanthurus, but below those of high-energy-demand species such as tunas Thunnus spp. and dolphinfish Coryphaena hippurus. Repeated-measures non-linear mixed-effects models were applied to account for within-individual autocorrelation and corrected for non-constant variance typical of noisy R(A) data sets. Repeated-measures models incorporating autoregressive first-order [AR(1)] and autoregressive moving average (ARMA) covariances provided significantly superior fits, more precise parameter estimates (i.e. reduced s.e.) and y-intercept estimates that more closely approximated measured R(R) for M. undulatus and L. xanthurus than standard least-squares regression procedures. 相似文献
58.
59.
William C Hamlett John A Musick Christopher K Hysell David M Sever 《The Journal of experimental zoology》2002,292(2):129-144
The fate of spermatozoa deposited within the female reproductive tract has been described in the smoothhound, Mustelus canis. Evidence of uterine epithelial-sperm interaction is presented, as well as documentation of sperm storage specifically in the terminal zone of the oviducal gland. Sperm fate is correlated with morphology of the endometrial cycle and specificity of storage in the oviducal gland. The endometrium of M. canis undergoes dramatic tissue remodeling associated with gestation. In females harboring fertilized ova or preimplantation yolk-reliant embryos, the uterine epithelium is simple cuboidal with mucous droplets for lubrication. The presence of the embryo elicits a response from the uterus, which becomes modified for nutrient and respiratory exchange into vascular uterine attachment sites that abut the distal aspect of the yolk sac. Areas of the uterus adjacent to the uterine attachment sites are termed paraplacental sites. Uterine attachment sites are simple squamous while the paraplacental epithelium is simple columnar. Paraplacental cells have basal metachromatic vesicles and a dense array of apical cytoplasmic filaments. Immediately postpartum the uterine attachment sites, now termed uterine or placental scars, begin to remodel to a mucous epithelium for the next gestational cycle. Paraplacental cells slough off the apical filamentous portion, and sperm become embedded in the epithelium. Bundled sperm occur throughout gestation in the terminal zone of the oviducal gland. Sperm are not embedded in the terminal zone epithelium as in the uterus. Following sperm release from the uterus, the paraplacental epithelium reverts to a mucous epithelium for the next reproductive cycle. Fertilization is presumed to occur in the anterior oviduct above the oviducal gland. The physiological mechanisms that mediate sperm-uterus attachment, release, and storage in the terminal zone of the oviducal gland are currently under investigation. 相似文献
60.
Human alveolar macrophage fibronectin: synthesis, secretion, and ultrastructural localization during gelatin-coated latex particle binding 总被引:10,自引:4,他引:10 下载免费PDF全文
Human pulmonary alveolar macrophages synthesized and secreted several characteristic high molecular weight proteins for at least 7 d in vitro. Immunoprecipitates of medium and cell lysates from metabolically labeled cultures with specific anti-human plasma fibronectin IgG contained one major labeled polypeptide of molecular weight 440,000 (unreduced) or 220,000 (reduced). An identical polypeptide in conditioned medium from radiolabeled macrophages bound specifically to gelatin-Sepharose, demonstrating that alveolar macrophages synthesized and secreted a molecule immunologically and functionally similar to fibronectin. Fibronectin was the major newly synthesized and secreted polypeptide of freshly harvested alveolar macrophages. Pulse-chase experiments revealed that newly synthesized fibronectin was rapidly secreted into medium, approximately 50 percent appearing by 1 h and 80 percent by 8 h. Immunoperoxidase staining using antifibronectin F(ab’)(2)-peroxidase conjugates revealed the majority of immunoreactive fibronectin to be intracellular, localized to endoplasmic reticulum and Golgi apparatus. No extracellular matrix fibronectin was visualized, and cell surface staining was rarely seen, usually appearing only at sites where cells were closely apposed and not at sites of macrophage-substrate attachment. Similar immunostaining of fibroblast cultures revealed cell surface-associated fibrillar fibronectin. Ultrastructural localization of fibronectin during binding and phagocytosis of gelatin-coated and plain latex particles revealed fibronectin only on gelatin-latex beads and at their cell binding sites. Neigher plain latex beads nor their cell membrane binding sites stained for fibronectin. These results demonstrate that fibronectin is a major product of human alveolar macrophages, is rapidly secreted, and is localized at cell membrane binding sites for gelatin-coated particles. In view of the known binding properties of fibronectin, it may serve as an endogenous opsonic factor promoting the binding of staphylococcus, denatured collagen, fibrin, or other macromolecules to macrophages in the lower respiratory tract. 相似文献