Developmental differences in genome replication program and origin activation

To ensure error-free duplication of all (epi)genetic information once per cell cycle, DNA replication follows a cell type and developmental stage specific spatio-temporal program. Here, we analyze the spatio-temporal DNA replication progression in (un)differentiated mouse embryonic stem (mES) cells. Whereas telomeres replicate throughout S-phase, we observe mid S-phase replication of (peri)centromeric heterochromatin in mES cells, which switches to late S-phase replication upon differentiation. This replication timing reversal correlates with and depends on an increase in condensation and a decrease in acetylation of chromatin. We further find synchronous duplication of the Y chromosome, marking the end of S-phase, irrespectively of the pluripotency state. Using a combination of single-molecule and super-resolution microscopy, we measure molecular properties of the mES cell replicon, the number of replication foci active in parallel and their spatial clustering. We conclude that each replication nanofocus in mES cells corresponds to an individual replicon, with up to one quarter representing unidirectional forks. Furthermore, with molecular combing and genome-wide origin mapping analyses, we find that mES cells activate twice as many origins spaced at half the distance than somatic cells. Altogether, our results highlight fundamental developmental differences on progression of genome replication and origin activation in pluripotent cells.     

Synthesis and pharmacological evaluation of bicyclic SNC80 analogues with separated benzhydryl moiety

Directed by molecular modeling studies the pharmacophoric benzhydryl moiety of the delta opioid receptor agonist SNC80 was separated and the two phenyl residues were attached to different positions of the conformationally constrained 6,8-diazabicyclo[3.2.2]nonane framework in order to find novel delta agonists. The crucial reaction step in the chiral pool synthesis was the establishment of the three carbon bridge by a Dieckmann analogous cyclization of the allyl and propyl derivatives 6 and 7 to yield the mixed methyl silyl acetals 8 and 9, respectively. Stereoselective Grignard reaction, dehydration, and introduction of the pharmacophoric (N,N-diethylcarbamoylbenzyl) residue led to the designed delta receptor agonists 3, ent-3, and 20 with a double bond in the bicyclic framework. Hydrogenation of the allyl derivative 14 was performed with ammonium formate and Pd/C to yield the saturated ligands 24a and 24b. Removal of the allyl substituent with RhCl(3), hydrogenation of the ring system, and re-attachment of the allyl moiety provided the allyl derivatives 4a and 4b. In receptor binding studies with the radioligand [(3)H]-deltorphine II only ent-3 showed considerable delta receptor affinity (K(i)=740 nM). Since ent-3 also interacts with mu receptors (K(i)=250 nM) it belongs to the very interesting compound class of mixed delta/mu ligands.     

Power and maximum strength relationships during performance of dynamic and static weighted jumps

The purpose of this study was to investigate the relationship of the 1 repetition maximum (1RM) squat to power output during countermovement and static weighted vertical squat jumps. The training experience of subjects (N = 22, 87.0 +/- 15.3 kg, 14.1 +/- 7.1% fat, 22.2 +/- 3.8 years) ranged from 7 weeks to 15+ years. Based on the 1RM squat, subjects were further divided into the 5 strongest and 5 weakest subjects (p 相似文献   

A cybernetic approach to osteoporosis in anorexia nervosa

A group of 25 female individuals, who had been admitted to the University Hospital with the diagnosis of anorexia nervosa (AN) 3 to 10 years before, was seen for a follow-up visit in the hospital. These women got a psychiatric exploration to detect a present eating disorder. Moreover, parameters of the muskuloskeletal interaction were determined on the non-dominant forearm. Bone mineral content (BMC) of the radius was measured by pQCT and maximal grip force was evaluated by the use of a dynamometer. Eating disorders were present in 12 females. The mean of BMC standard deviation (SD) score was significantly reduced in comparison with reference values. Furthermore, the mean of BMC SD score was also significantly lower than the mean of grip force in SD score. These results gave the suggestion that the adaptation of bone mass to biomechanical forces is disturbed in AN. The linear regression analyses between the parameters grip force and BMC were compared between the study and the reference group. The comparison delivered a significantly lower constant in the regression equation of the study group. This result can be interpreted on the background of the mechanostat theory. The affection with an eating disorder decreases the set point in the feedback loop of bone modeling. The results offer for the first time the possibility to analyse osteoporosis in anorexic females under the paradigm of muskuloskeletal interaction.     

Calves born after open pulled straw vitrification of immature bovine oocytes

The aim of this study was to evaluate the developmental capacity of immature bovine oocytes after vitrification with 20% ethylene glycol (EG)+20% dimethyl sulfoxide (Me(2)SO) and 0.5M sucrose (SUC), by open pulled straw (OPS) technology. The effect of treatment with cytochalasin D before vitrification was also examined. No differences were observed in cleavage and blastocyst rates among the group vitrified without cytochalasin D treatment (Vitri) (49.0% and 6.1%) and that with cytochalasin D treatment before vitrification (CDVitri) (46.4% and 3.6%), but both were lower (P<0.05) than the unvitrified control group (85.1 and 45.9%). Calves were obtained after transfer of fresh and vitrified blastocysts from the Vitri group and after transfer of vitrified blastocysts from the CDVitri group. Cytochalasin D treatment does not improve the development of immature bovine vitrified oocytes. The results show that a small proportion of immature oocytes vitrified with this technology are fully competent to produce blastocysts, which may be transferred immediately or vitrified before transfer, and go on to develop healthy offspring.     

Inferring the number of evolutionary events from DNA coding sequence differences

The estimation of the amount of evolutionary divergence that has taken place between two DNA coding sequences depends strongly on the degree of constraint on amino acid replacements. If amino acid replacements are relatively unconstrained, the individual nucleotide is the appropriate unit of analysis and the method of Tajima and Nei can be used. If amino acid replacements are constrained, however, this method is shown to be inapplicable. For sequences with strong amino acid constraints, a method is outlined analogous to the Tajima and Nei method using codons as the unit of analysis. Only synonymous substitutions are used. Codon usage data can be employed to estimate the necessary parameters of the calculation, or a priori models of substitution may be employed. Sequences with significant but intermediate constraints on amino acid replacements are, in principle, unanalyzable.      

Mitogenic activity of pituitary hormones on cell cultures of normal and carcinogen-induced tumor epithelium from rat mammary glands

Cell suspension containing normal or tumor epithelium were readily obtained by enzymatically digesting rat mammary glands from perphenazine-treated (prolactin-hypersecreting) cycling, female virgin animals or hormone- responsive mammary tumors from animal treated with dimethylbenzanthracene. Cell suspensions were fractioned into predominantly epithelial and predominantly stromal cells by their differential rates of attachment to culture dishes. Both normal mammary and tumor epithelial cells were characterized by the presence of specific cell-junctional complexes, desmosome-like structures, surface microvilli, and their ability to synthesize casein. Serum-dependent protease activity was greater in cultures derived from tumors, and cells from such cultures grew in agarose whereas those from the non-neoplastic gland did not. The addition of prolactin to the culture medium stimulated DNA synthesis in primary or secondary epithelial cultures from tumors, whereas additional insulin and hydrocortisone with prolactin were required for similar levels of DNA synthesis in cultures from non-neoplastic glands. The fraction of cells synthesizing DNA was, however, smaller than that with 10 percent serum measured in the same time period. Both growth hormone and epidermal growth factor stimulated DNA synthesis but to a lesser extent than did prolactin. Prolactin with hydrocortisone and insulin were relatively inactive in promoting DNA synthesis of the nonepithelial cells whereas pituitary fibroblast growth factor was more active. These mitogenic effects were obtained when the hormones were added to the medium at near physiological concentrations, and paralleled the known activities of the hormones in control of mammary gland growth and development in the rat.     

Climatic change in the Russian Altai, southern Siberia, based on palynological and geomorphological results, with implications for climatic teleconnections and human history since the middle Holocene

Two pollen diagrams from near the Chuya Basin, Russian Altai, are presented together with results from geomorphological archives. The Kuray Range profile (2330 m a.s.l.) is situated just above the forest line and starts with the weak cooling of the middle Atlantic period (ca. 6500–5900 b.p.) that bisects the Holocene optimum, as in other parts of Siberia. Taiga (boreal coniferous forest) with Picea obovata established afterwards but was displaced at ca. 5300 b.p. by the pronounced cooling of the early Sub-boreal. A gradual recovery of taiga stopped and it disappeared around 3400 b.p. at the end of the middle Sub-boreal. Since then the climatic and vegetational conditions much resemble those of the Sub-atlantic. In the last two millennia, three climatic declines are documented in the second diagram from the Tarkhata Valley (2210 m a.s.l.) from the dry limit of Larix sibirica: a cold and wet one around the 5th century a.d., a cold and dry one around a.d. 1200 and the Little Ice Age which started around a.d. 1600. In addition, several of the climatic periods and especially local human influence can be proved by the geomorphological findings. Most of the climatic changes seem to be connected with socio-ecological changes in Asia and even Europe, with movements of the Huns and Mongols, which show the possible dimensions of global climatic change. Hints of teleconnections between Siberia, the Mediterranean and the Himalayas via the North Atlantic and the Arctic Oscillation are discussed.     

Short-Term Response of Land Birds to Ponderosa Pine Restoration

We monitored the short‐term (>3 years) response of land birds to forest restoration treatments in Ponderosa pine forests located on the east slope of the North Cascade Range. Restoration treatments were designed to create stand structure and composition similar to pre‐settlement forests, which were influenced by a frequent fire regime. Overall, avian community composition was changed as a result of the treatments. Cassin’s Finch, Chipping Sparrow, and Yellow‐rumped Warbler were found at higher densities in treated stands, whereas Mountain Chickadee, Western Tanager, and Red‐breasted Nuthatch had higher densities in untreated stands. White‐headed Woodpecker and Western Bluebird were only detected in the treated stands. Brown‐headed Cowbird showed no response to treatments. We detected changes in the density of four of five foraging guilds in response to restoration treatments. Tree seedeaters, low understory and ground insectivores, and aerial insectivores all increased in density in treated stands. Overall, bark insectivores showed no density response to treatments. Tree foliage insectivore density was lower in treated than in untreated stands. Overall avian density, density of neotropical migrants, and density of some focal species were higher in treated stands. Monitoring should be continued to understand the longer‐term (5–10 year) responses of land birds and to guide future forest restoration efforts.