Structure of DNMT1-DNA complex reveals a role for autoinhibition in maintenance DNA methylation

Maintenance of genomic methylation patterns is mediated primarily by DNA methyltransferase-1 (DNMT1). We have solved structures of mouse and human DNMT1 composed of CXXC, tandem bromo-adjacent homology (BAH1/2), and methyltransferase domains bound to DNA-containing unmethylated CpG sites. The CXXC specifically binds to unmethylated CpG dinucleotide and positions the CXXC-BAH1 linker between the DNA and the active site of DNMT1, preventing de novo methylation. In addition, a loop projecting from BAH2 interacts with the target recognition domain (TRD) of the methyltransferase, stabilizing the TRD in a retracted position and preventing it from inserting into the DNA major groove. Our studies identify an autoinhibitory mechanism, in which unmethylated CpG dinucleotides are occluded from the active site to ensure that only hemimethylated CpG dinucleotides undergo methylation.     

Ethylene modulates stem cell division in the Arabidopsis thaliana root

The construction of multicellular organisms depends on stem cells-cells that can both regenerate and produce daughter cells that undergo differentiation. Here, we show that the gaseous messenger ethylene modulates cell division in the cells of the quiescent center, which act as a source of stem cells in the seedling root. The cells formed through these ethylene-induced divisions express quiescent center-specific genes and can repress differentiation of surrounding initial cells, showing that quiescence is not required for these cells to signal to adjacent stem cells. We propose that ethylene is part of a signaling pathway that modulates cell division in the quiescent center in the stem cell niche during the postembryonic development of the root system.     

Constraints on neon and argon isotopic fractionation in solar wind

To evaluate the isotopic composition of the solar nebula from which the planets formed, the relation between isotopes measured in the solar wind and on the Sun's surface needs to be known. The Genesis Discovery mission returned independent samples of three types of solar wind produced by different solar processes that provide a check on possible isotopic variations, or fractionation, between the solar-wind and solar-surface material. At a high level of precision, we observed no significant inter-regime differences in 20Ne/22Ne or 36Ar/38Ar values. For 20Ne/22Ne, the difference between low- and high-speed wind components is 0.24 +/- 0.37%; for 36Ar/38Ar, it is 0.11 +/- 0.26%. Our measured 36Ar/38Ar ratio in the solar wind of 5.501 +/- 0.005 is 3.42 +/- 0.09% higher than that of the terrestrial atmosphere, which may reflect atmospheric losses early in Earth's history.     

Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum

Flow cytometry is an essential tool for dissecting the functional complexity of hematopoiesis. We used single-cell "mass cytometry" to examine healthy human bone marrow, measuring 34 parameters simultaneously in single cells (binding of 31 antibodies, viability, DNA content, and relative cell size). The signaling behavior of cell subsets spanning a defined hematopoietic hierarchy was monitored with 18 simultaneous markers of functional signaling states perturbed by a set of ex vivo stimuli and inhibitors. The data set allowed for an algorithmically driven assembly of related cell types defined by surface antigen expression, providing a superimposable map of cell signaling responses in combination with drug inhibition. Visualized in this manner, the analysis revealed previously unappreciated instances of both precise signaling responses that were bounded within conventionally defined cell subsets and more continuous phosphorylation responses that crossed cell population boundaries in unexpected manners yet tracked closely with cellular phenotype. Collectively, such single-cell analyses provide system-wide views of immune signaling in healthy human hematopoiesis, against which drug action and disease can be compared for mechanistic studies and pharmacologic intervention.     

Allometric equations for aboveground biomass estimation by size class for Pinus brutia Ten. trees growing in North and South Aegean Islands, Greece

Empirical allometric equations relating biomass of aboveground components to dendrometric variables for Pinus brutia Ten. trees are derived in this paper. They are based on data collected from Lesvos (North Aegean Sea) and Crete (South Aegean Sea) Islands. Comparisons to published equations for the same species growing in northwestern and southeastern Turkey, for Pinus nigra A. growing in Turkey and Pinus halepensis Mill. found in Western Aegean (island of Evia), are also presented. The biomass of branches from destructively sampled trees (twelve in Crete and six in Lesvos) was divided into four size classes (0?C0.63 cm, 0.64?C2.5 cm, 2.51?C7.61 cm, and 7.62?C22.8 cm). Tree crown biomass was calculated as the sum of the biomass in the four classes plus the fraction of stem above crown base. Over bark stem biomass was estimated through bole volume conversion based on wood density. The results showed clearly that, for a given diameter, the Cretan trees had more crown biomass and a higher share of small branches than trees on Lesvos, probably due to differences in environment and stand structure. Comparisons to published diameter versus crown biomass equations reveal a lower crown biomass for Turkish sites of Calabrian pine and Aleppo pine on Evia Island, while only Turkish Black pine seems to be comparable to the Calabrian pine on Crete. The derived allometries can be used for landscape fire behavior modeling, for ecophysiological studies and for the Kyoto protocol requirements of carbon changes in Pinus brutia Ten. forests located in northern and southern Greek sites.     

The examination of biophysical parameters of skin (transepidermal water loss, skin hydration and pH value) in different body regions of normal cats of both sexes

The purpose of this study was to evaluate transepidermal water loss (TEWL), skin hydration and skin pH in normal cats. Twenty shorthaired European cats of both sexes were examined in the study. Measurements were taken from five different sites: the lumbar region, the axillary fossa, the inguinal region, the ventral abdominal region and the left thoracic region. In each of the regions, TEWL, skin hydration and skin pH were measured. The highest TEWL value was observed in the axillary fossa (18.22g/h/m(2)) and the lowest in the lumbar region (10.53g/h/m(2)). The highest skin hydration was found in the inguinal region (18.29CU) and the lowest in the lumbar region (4.62CU). The highest skin pH was observed in the inguinal region (6.64) and the lowest in the lumbar region (6.39). Statistically significant differences in TEWL were observed between the lumbar region and the left side of the thorax region (P=0.016), the axillary fossa (P=0.0004), the ventral region (P=0.005), and the inguinal region (P=0.009). There were significant differences in skin hydration between the lumbar region and the left thorax (P=0.000003), the axillary fossa (P=0.002), the ventral abdomen (P=0.03), and the inguinal region (P=0.0003) as well as between the thorax and the ventral abdomen (P=0.005). TEWL was higher in females (15g/h/m(2)) than in males (4.57g/h/m(2)). Skin hydration was higher in females (13.89CU) than in males (12.28CU). Significant differences were not found between males and females for TEWL and skin hydration. Skin pH was higher in males (6.94) than in females (6.54), which was significant (P=0.004).     

Local immunity of the respiratory mucosal system in chickens and turkeys

This review article presents fundamental mechanisms of the local mucosal immunity in selected regions of the respiratory tract in healthy birds and in some pathological conditions. The respiratory system, whose mucosa come into direct contact with microorganisms contaminating inhaled air, has some associated structures, such as Harderian gland (HG), conjunctive-associated lymphoid tissue (CALT) and paranasal glands (PG), whose participation in local mechanisms of the mucosal immunity has been corroborated by numerous scientific studies. The nasal mucosa, with structured clusters of lymphoid tissue (NALT - nasal-associated lymphoid tissue) is the first to come into contact with microorganisms which contaminate inhaled air. Lymphoid nodules, made up of B cells with frequently developed germinal centres (GC), surrounded by a coat of CD4+ cells, are the major NALT structures in chickens, whereas CD8+ cells are situated in the epithelium and in the lamina propria of the nasal cavity mucosa. Studies into respiratory system infections (e.g. Mycoplasma gallisepticum) have shown the reactivity of the tracheal mucosa to infection, despite a lack of essential lymphoid tissue. Bronchus-associated lymphoid tissue (BALT) takes part in bronchial immune processes and its structure, topography and ability to perform defensive function in birds is largely age-dependent. Mature BALT is covered by a delicate layer of epithelial cells, called follicle-associated epithelium (FAE). Germinal centres (GC), surrounded by CD4+ cells are developed in most mature BALT nodules, while CD8+ lymphocytes are dispersed among lymphoid nodules and in the epithelium, and they are rarely present in GC. Macrophages make up the first line of defence mechanisms through which the host rapidly responds to microorganisms and their products in the respiratory mucosal system. Another very important element are polymorphonuclear cells, with heterophils being the most important of them. Phagocytic cells obtained from lung lavages in birds are referred to as FARM (free avian respiratory macrophages). Their number in chickens and turkeys is estimated to be 20 times lower than that in mice and rats, which indicates a deficit in the first-line of defence in the birds' respiratory system. There are numerous B cells and antibody secreting cells (ASC) present throughout the respiratory system in birds. Their role comes down to perform antigen-specific protection by producing antibodies (IgM, IgY or IgA class) as a result of contact with pathogenic factors.     

Real-time PCR detection of Mycoplasma felis in domestic cats suffering from chronic conjunctivitis (Poland)

Real-time PCR directed to intergenic spacer (IGS) noncoding region between 16S and 23S rRNA genes was used for species specific detection of Mycoplasma felis in conjunctival scrapings. Samples were collected from 57 cats suffering from chronic conjunctivitis in 2008-2010 (Wroc?aw, Poland). Samples from 36 cats (63.2%) were shown to be positive for Mycoplasma felis. Our research gives a first insight in the occurrence of Mycoplasma felis among domestic cats in Poland suggesting that this pathogen may constitute an underestimated cause of chronic conjunctivitis.     

Role of extracellular matrix and prolactin in functional differentiation of bovine BME-UV1 mammary epithelial cells

Interactions between extracellular matrix (ECM) and epithelial cells are necessary for proper organisation and function of the epithelium. In the present study we show that bovine mammary epithelial cell line BME-UV1 cultured on ECM components, commercially available as Matrigel, constitutes a good model for studying mechanisms controlling functional differentiation of the bovine mammary gland. In contact with Matrigel BME-UV1 cells induce apicobasal polarity, and within 16 days form three dimensional (3D) acinar structures with a centrally localized hollow lumen, which structurally resemble mammary alveoli present in the functionally active mammary gland. We have shown that the 3D culture system enables a high expression and proper localisation of integrin receptors and tight junction proteins in BME-UV1 cells to be induced. This effect was not obtained in cells grown in the classical 2D culture system on plastic. Moreover, ECM highly stimulated the synthesis of one of the major milk proteins, beta-casein, even in the absence of prolactin. Our results show that contact with ECM plays an important role in the lactogenic activity of bovine MECs, however, prolactin is necessary for the efficient secretion of milk proteins.