稀土多元复合肥和三种稀土元素的遗传毒性研究

采用蚕豆根尖细胞微核技术,研究市售稀土多元复合肥和稀土元素镧、铈、铒的化合物对蚕豆根尖细胞的遗传毒性和细胞毒性。结果表明,稀土复合肥和三种稀土元素均可诱发微核效应,在一定浓度下可损伤细胞,影响根尖的正常生长,其中稀土复合肥的微核效应表现出明显的剂量-效应关系。稀土复合肥和稀土元素镧、铈、铒的化合物对蚕豆根尖细胞具有一定的遗传毒性作用和细胞毒性作用,在施用稀土微肥和使用稀土制品时应引起重视。 Abstract：This paper presents the study of genetic toxicity and cell toxicity that is give n by rare earth multi-element compound fertilizer and a chemical compound of rar e earth elements-La3＋、Ce4＋、Er3＋ in root tip cells of Vicia faba.The technique used is micronucleus in root tip cells of Vicia faba.The experiment statistical result shows that both the rare earth compound fertilizer and the three kinds of rare earth elements can cause micronucleus ef fect and under certain concentration,they can hurt cells, affect root tip gro wth .The micronucleus effect of the rare earth compound fertilizer shows a clear relation of dosage-effect. The conclusion is that rare earth compound fertilize r and the chemical compound of rare earth elements La3＋、Ce4＋、E r3＋cause certain genetic toxicity and cell toxicity effect to root tip cells of Vicia faba.Therefore a close attention should be paid when the rar e earth multi-fertilizer and other things made by rare earth are used.     

Dynamics of granzyme B-induced apoptosis: mathematical modeling

Apoptosis is mediated by an intracellular biochemical system that mainly includes proteins (procaspases, caspases, inhibitors, Bcl-2 protein family as well as substances released from mitochondrial intermembrane space). The dynamics of caspase activation and target cleavage in apoptosis induced by granzyme B in a single K562 cell was studied using a mathematical model of the dynamics of granzyme B-induced apoptosis developed in this work. Also the first application of optimization approach to determination of unknown kinetic constants of biochemical apoptotic reactions was presented. The optimization approach involves solving of two problems: direct and inverse. Solving the direct optimization problem, we obtain the initial (baseline) concentrations of procaspases for known kinetic constants through conditional minimization of a cost function based on the principle of minimum protein consumption by the apoptosis system. The inverse optimization problem is aimed at determination of unknown kinetic constants of apoptotic biochemical reactions proceeding from the condition that the optimal concentrations of procaspases resulting from the solution of the direct optimization problem coincide with the observed ones, that is, those determined by biochemical methods. The Multidimensional Index Method was used to perform numerical solution of the inverse optimization problem.     

Lanthanum enhances in vitro osteoblast differentiation via pertussis toxin-sensitive gi protein and ERK signaling pathway

Converging lines of evidence suggest that lanthanum tends to deposit in bone. The influence of lanthanum ion (La3+) on osteoblast differentiation and the related mechanism are essential to understanding its effect on bone metabolism. In this study, La3+ treatment enhanced in vitro osteoblast differentiation as evidenced by promoting alkaline phosphatase (ALP) activity, osteocalcin (OC) secretion, and matrix mineralization. The expressions of osteoblast-specific genes of Cbfa-1, osteopontin (OPN), and bone sialoprotein (BSP) were all increased in the presence of La3+, but no change was observed in that of type I collagen (COL-I). Further studies demonstrated that La3+ treatment enhanced phosphorylation of extracellular signal-regulated kinase (ERK). Inhibition of ERK activation by U0126 suppressed the effects of La3+ on osteoblast activity. Moreover, pretreatment of the cells with pertussis toxin (PTx), a Gi protein inhibitor, suppressed the La3+-enhanced ERK phosphorylation and osteoblast differentiation. These findings suggest that La3+ exposure enhances in vitro osteoblast differentiation and the effect depends on ERK phosphorylation via PTx-sensitive Gi protein signaling.     

Organic acids promote the uptake of lanthanum by barley roots

Organic acids play an important role in metal uptake by, and accumulation in, plants. However, the relevant mechanisms remain obscure. Acetic, malic and citric acids increased the uptake of lanthanum (La) by barley (Hordeum vulgare) roots and enhanced La content in shoots under hydroponic conditions. Concentration-dependent net La influx in the absence and presence of organic acids yielded nonsaturating kinetic curves that could be resolved into linear and saturable components. The saturable component followed Michaelis-Menten kinetics. The K(m) values were similar; however, the V(max) values in the presence of acetic, malic and citric acids were 4.3, 2.8, 1.5-times that of the control, respectively. Enhanced uptake of La by organic acids was mediated mainly, but not solely, by Ca(2+) channels. X-ray absorption spectroscopic techniques provided evidence of La-oxygen environment and established that La(III) was coordinated to 11 oxygen atoms that are likely to be involved in the binding of La(III) to barley roots via carboxylate groups and hydration of La(III).     

La, PTB, and PAB proteins bind to the 3(') untranslated region of Norwalk virus genomic RNA

Noroviruses are human enteric caliciviruses for which no cell culture is available. Consequently, the mechanisms and factors involved in their replication have been difficult to study. In an attempt to analyze the cis- and trans-acting factors that could have a role in NV replication, the 3(')-untranslated region of the genome was studied. Use of Zuker's mfold-2 software predicted that NV 3(')UTR contains a stem-loop structure of 47 nts. Proteins from HeLa cell extracts, such as La and PTB, form stable complexes with this region. The addition of a poly(A) tail (24 nts) to the 3(')UTR permits the specific binding of the poly(A) binding protein (PABP) present in HeLa cell extracts, as well as the recombinant PABP. Since La, PTB, and PABP are important trans-acting factors required for viral translation and replication, these RNA-protein interactions may play a role in NV replication or translation.     

Coordinated assembly of Ku and p460 subunits of the DNA-dependent protein kinase on DNA ends is necessary for XRCC4-ligase IV recruitment

Repair of DNA double-strand breaks by the non-homologous end-joining pathway (NHEJ) requires a minimal set of proteins including DNA-dependent protein kinase (DNA-PK), DNA-ligase IV and XRCC4 proteins. DNA-PK comprises Ku70/Ku80 heterodimer and the kinase subunit DNA-PKcs (p460). Here, by monitoring protein assembly from human nuclear cell extracts on DNA ends in vitro, we report that recruitment to DNA ends of the XRCC4-ligase IV complex responsible for the key ligation step is strictly dependent on the assembly of both the Ku and p460 components of DNA-PK to these ends. Based on co-immunoprecipitation experiments, we conclude that interactions of Ku and p460 with components of the XRCC4-ligase IV complex are mainly DNA-dependent. In addition, under p460 kinase permissive conditions, XRCC4 is detected at DNA ends in a phosphorylated form. This phosphorylation is DNA-PK-dependent. However, phosphorylation is dispensable for XRCC4-ligase IV loading to DNA ends since stable DNA-PK/XRCC4-ligase IV/DNA complexes are recovered in the presence of the kinase inhibitor wortmannin. These findings extend the current knowledge of the assembly of NHEJ repair proteins on DNA termini and substantiate the hypothesis of a scaffolding role of DNA-PK towards other components of the NHEJ DNA repair process.     

Differential phenotypic variability among the Apalachee mission populations of La Florida: a diachronic perspective

Phenotypic variability is evaluated in a series of skeletal samples from the Apalachee region of Florida. Based on ethnohistoric evidence, several predictive models for changes in variability are generated. If variability decreases through time, this likely represents the effect of genetic drift in populations experiencing epidemic disease and population loss. If variability increases through time, this suggests that population aggregation or genetic admixture were primary factors shaping the Apalachee population during the mission period. Dental dimensions were collected from a series of precontact (pre-1500), early mission (AD 1633-1650) (San Pedro y San Pablo de Patale), and late mission (post-1657) (San Luis) samples from the Apalachee region and were subjected to univariate and multivariate variability analyses. The results indicate that the late mission San Luis sample was significantly more variable than the Patale or precontact samples; however, the Patale sample exhibited no significant variability change in comparison to the precontact population. This suggests that the missions initially effected limited change in genetic variability in the mission populations. However, San Luis was affected by either admixture or population aggregation to such a degree that the observed variation had increased beyond earlier levels. Given the limited historic evidence for population aggregation at this mission, and the comparatively large resident Spanish population, the increased variability may be indicative of admixture at this mission, and potentially at this mission only. Based on a limited data set, however, it appears that the mission period cannot be typified by a single evolutionary or historic process.     

Effect of cysteine,glutamate and glycine supplementation to in vitro fertilization medium during bovine early embryo development

Glutathione (GSH) is an antioxidant synthesized from three constitutive amino acids (CAA): cysteine (Cys), glycine (Gly) and glutamate (Glu). Glutathione plays an important role in oocyte maturation, fertilization and early embryo development. This study aimed to investigate the effect of Cys (0.6 mM), Gly (0.6 mM) and Glu (0.9 mM) supplementation during in vitro fertilization (IVF) of cattle oocytes. In a Pilot Experiment, de novo synthesis of GSH in bovine zygote was evaluated using a modified TALP medium prepared without MEM-essential and MEM-non-essential amino acids (mTALP): mTALP + CAA (constitutive amino acids); mTALP + CAA+5 mMBSO (buthionine sulfoximide); mTALP + Cys + Gly; mTALP + Cys + Glu and mTALP + Gly + Glu. This evidence led us to investigate the impact of CAA supplementation to TALP medium (with essential and non-essential amino acids) on zygote viability, lipid peroxidation, total intracellular GSH content (include reduced and oxidized form; GSH-GSSG), pronuclear formation in zygotes and subsequent embryo development. IVF media contained a) TALP; b) TALP + Cys + Gly + Glu (TALP + CAA); c) TALP + Cys + Gly; d) TALP + Cys + Glu; e) TALP + Gly + Glu, were used. Total GSH-GSSG concentration was increased in TALP, TALP + CAA, and TALP + Cys + Gly. The viability of zygote was similar among treatments. Lipid peroxidation was increased in zygote fertilized with TALP + Cys + Gly; TALP + Cys + Glu; TALP + Gly + Glu and TALP + CAA. The percentage of penetrated oocytes decreased in TALP + CAA and TALP + Cys + Gly. The cleavage rate was lower in TALP + CAA and TALP + Gly + Glu. The percentage of embryos developing to the blastocyst stage was lower in TALP + Cys + Glu and TALP + CAA. In conclusion, we have demonstrated the synthesis of GSH during IVF. However, Cys, Gly and Glu supplementation to TALP medium had negative effects on embryonic development.     

Attempted DNA extraction from a Rancho La Brea Columbian mammoth (Mammuthus columbi): prospects for ancient DNA from asphalt deposits

Fossil‐bearing asphalt deposits are an understudied and potentially significant source of ancient DNA. Previous attempts to extract DNA from skeletons preserved at the Rancho La Brea tar pits in Los Angeles, California, have proven unsuccessful, but it is unclear whether this is due to a lack of endogenous DNA, or if the problem is caused by asphalt‐mediated inhibition. In an attempt to test these hypotheses, a recently recovered Columbian mammoth (Mammuthus columbi) skeleton with an unusual pattern of asphalt impregnation was studied. Ultimately, none of the bone samples tested successfully amplified M. columbi DNA. Our work suggests that reagents typically used to remove asphalt from ancient samples also inhibit DNA extraction. Ultimately, we conclude that the probability of recovering ancient DNA from fossils in asphalt deposits is strongly (perhaps fatally) hindered by the organic compounds that permeate the bones and that at the Rancho La Brea tar pits, environmental conditions might not have been ideal for the general preservation of genetic material.