人骨髓基质细胞输注裸鼠后的体内分布

采用密度梯度离心和差速贴壁法分离培养人骨髓基质细胞.对细胞进行形态观察、表型分析、细胞生长曲线和成骨分化潜能测定.然后将第4代细胞通过尾静脉输注到未经任何处理的裸鼠体内,移植1周和4周后,利用RT-PCR检测人Alu基因在裸鼠不同脏器组织中的表达以判断人源性供体细胞的植入情况.结果表明培养的人骨髓基质细胞为CD34 -CD45-成纤维样细胞,并具有向成骨分化的能力,但第1至第5代人骨髓基质细胞增殖速度无显著性差异(P＞0.05).经尾静脉输注后,人源性供体细胞可在肺、肝、脾、肾、肌肉、心、肠、脑等多种脏器或组织中均有不同程度的分布.     

The HIV-1 p6/EIAV p9 docking site in Alix is autoinhibited as revealed by a conformation-sensitive anti-Alix monoclonal antibody

Alix [ALG-2 (apoptosis-linked gene 2)-interacting protein X], a component of the endosomal sorting machinery, contains a three-dimensional docking site for HIV-1 p6(Gag) or EIAV (equine infectious anaemia virus) p9(Gag), and binding of the viral protein to this docking site allows the virus to hijack the host endosomal sorting machinery for budding from the plasma membrane. In the present study, we identified a monoclonal antibody that specifically recognizes the docking site for p6(Gag)/p9(Gag) and we used this antibody to probe the accessibility of the docking site in Alix. Our results show that the docking site is not available in cytosolic or recombinant Alix under native conditions and becomes available upon addition of the detergent Nonidet P40 or SDS. In HEK (human embryonic kidney)-293 cell lysates, an active p6(Gag)/p9(Gag) docking site is specifically available in Alix from the membrane fraction. The findings of the present study demonstrate that formation or exposure of the p6(Gag)/p9(Gag) docking site in Alix is a regulated event and that Alix association with the membrane may play a positive role in this process.     

Identifying foldable regions in protein sequence from the hydrophobic signal

Structural genomics initiatives aim to elucidate representative 3D structures for the majority of protein families over the next decade, but many obstacles must be overcome. The correct design of constructs is extremely important since many proteins will be too large or contain unstructured regions and will not be amenable to crystallization. It is therefore essential to identify regions in protein sequences that are likely to be suitable for structural study. Scooby-Domain is a fast and simple method to identify globular domains in protein sequences. Domains are compact units of protein structure and their correct delineation will aid structural elucidation through a divide-and-conquer approach. Scooby-Domain predictions are based on the observed lengths and hydrophobicities of domains from proteins with known tertiary structure. The prediction method employs an A*-search to identify sequence regions that form a globular structure and those that are unstructured. On a test set of 173 proteins with consensus CATH and SCOP domain definitions, Scooby-Domain has a sensitivity of 50% and an accuracy of 29%, which is better than current state-of-the-art methods. The method does not rely on homology searches and, therefore, can identify previously unknown domains.     

The presence of 9-cis-beta-carotene in cytochrome b(6)f complex from spinach.

Cytochrome b(6)f complex with stoichiometrically bound beta-carotene molecule was purified from spinach chloroplasts. The configuration of this beta-carotene was studied by reversed-phase HPLC and resonance Raman spectroscopy. Both the absorption spectrum of this beta-carotene in dissociated state and the Raman spectrum in native state can be unambiguously assigned to a 9-cis configuration. This finding is in contrast to the predominantly all-trans isomers commonly found in membranes and protein-pigment complexes of chloroplasts, suggesting that the 9-cis-beta-carotene is an authentic component and may have a unique structural and functional role in cytochrome b(6)f complex.     

Tumorhead, a Xenopus gene product that inhibits neural differentiation through regulation of proliferation

Tumorhead (TH) is a novel maternal gene product from Xenopus laevis containing several basic domains and a weak coiled-coil. Overexpression of wild-type TH resulted in increased proliferation of neural plate cells, causing expansion of the neural field followed by neural tube and craniofacial abnormalities. Overexpressed TH protein repressed neural differentiation and neural crest markers, but did not inhibit the neural inducers, pan-neural markers or mesodermal markers. Loss of function by injection of anti-TH antibody inhibited cell proliferation. Our data are consistent with a model in which tumorhead functions in regulating differentiation of the neural tissues but not neural induction or determination through its effect on cell proliferation.     

Reproduction during spaceflight by plants in the family Brassicaceae

Researchers report on studies of reproduction in Arabidopsis thaliana in space during during the Chromex-03 on STS-54, Chromex-04 on STS-51, and Chromex-05 on STS-68 missions. The obstacles to seed formation were related to carbon dioxide levels. Other experiments examined in flight pollination and seed production in Brassica rapa during parabolic flight, a 4-1/2 month stay on Mir, and on STS-87. During the Mir experiment, Brassica seeds were harvested from seeds sown in flight. The second generation seeds grew to produce new seeds that contained more starch and less protein and lipid when compared to ground control seeds.     

FADD is required for DR4- and DR5-mediated apoptosis: lack of trail-induced apoptosis in FADD-deficient mouse embryonic fibroblasts

TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) is a member of the tumor necrosis factor family that can kill a wide variety of tumor cells but not normal cells. TRAIL-induced apoptosis in humans is mediated by its receptors DR4 (TRAIL-R1) and DR5 (TRAIL-R2). What constitutes the signaling molecules downstream of these receptors, however, remains highly controversial. Using the FADD dominant negative molecule, several groups have reached different conclusions with respect to the role of FADD in TRAIL-induced apoptosis. More recently, using FADD-deficient (-/-) mouse embryonic fibroblasts, Yeh et al. (Yeh, W.-C., Pompa, J. L., McCurrach, M. E., Shu, H.-B., Elia, A. J., Shahinian, A., Ng, M., Wakeham, A., Khoo, W., Mitchell, K., El-Deiry, W. S., Lowe, S. W., Goeddel, D. V., and Mak, T. W. (1998) Science 279, 1954-1958) concluded that DR4 utilizes a FADD-independent apoptotic pathway. The latter experiment, however, involved transient overexpression, which often leads to nonspecific aggregation of death domain-containing receptors. To address this issue in a more physiological setting, we stably transfected mouse DR4/5, human DR4, or human DR5 into FADD(-/-) mouse embryonic fibroblast cells. We showed that FADD(-/-) MEF cells stably transfected with TRAIL receptors are resistant to TRAIL-mediated cell death. In contrast, TRAIL receptors stably transfected into heterozygous FADD(+/-) cells or FADD(-/-) cells reconstituted with a FADD retroviral construct are sensitive to the TRAIL cytotoxic effect. We conclude that FADD is required for DR4- and DR5-mediated apoptosis.     

Study on blood constitutive parameters in different blood constitutive equations

A method is proposed to study the steady blood constitutive equation (BCE). With the method several BCEs are tested and Quemada, Bi-exponent and K-L equations are found to be well in agreement with the hemorheological characteristics of human and canine blood and these equations may be well used in hemorheology and hemodynamics. In addition, the potential clinical applications of some blood constitutive parameters (BCPs) are discussed. It is found that the hematocrit can significantly affect the BCPs: K(A) = alpha2/alpha1, Q(A) = eta0/eta(infinity), R(A) = etaA/eta(e) - 1, which means relative viscosities and they may be related to RBC aggregative level. Moreover, we also show that the parameters are related to the RBC aggregative strength and level.     

Chain Correlation between Variables of Gas Exchange and Yield Potential in Different Winter Wheat Cultivars

Variables of gas exchange of flag leaves and grain yield potentials of five representative winter wheat (Triticum aestivum L.) cultivars varied greatly across different development stages under the same management and irrigation. The cultivars with high yield potential had higher net photosynthetic rate (P _N), PPFD (photosynthetic photon flux density) saturated photosynthetic rate (P _sat), stomatal conductance (g _s), and maximum apparent quantum yield of CO₂ fixation (_m,app) than those with low grain yield, but their dark respiration rate (R _D) and compensation irradiance (I _c) were remarkably lower. Compared with overall increase of yield potential of 71 % from low yield cultivars to high yield ones, P _N, P _sat, _m,app, and g _s were 13, 19, 57, and 32 % higher, respectively; but R _D and I _c decreased by 19 and 76 %, respectively. Such difference was evidently large during anthesis stage (e.g., P _N by 33 %), which indicated that this period could be the best for assisting further selection for better cultivars. However, transpiration rate (E) and water use efficiency (WUE) differed only little. At different development stages, especially at anthesis, P _N and P _sat were positively correlated with _m,app, g _s, and yield potential, and negatively correlated with R _D and I _c. Thus the high-yield-potential winter wheat cultivars possess many better characters in photosynthesis and associated parameters than the low-yield cultivars.