For identification of genes involved in embryogenesis in the model cereal rice, we have constructed a collection of cDNA libraries of well-defined stages of embryo development before, during and ...after organ differentiation. Here, we focus on the possible role of KNOX (maize Knotted1-like) class homeobox genes in regulation of rice embryogenesis. Three types of KNOX clones were identified in libraries of early zygotic embryos. Two of these, Oskn2 and Oskn3, encode newly described KNOX genes, whereas the third (Oskn1) corresponds to the previously described OSH1 gene. In situ hybridizations showed that during the early stages of embryo development, all three KNOX genes are expressed in the region where the shoot apical meristem (SAM) is organizing, suggesting that these genes are involved in regulating SAM formation. Whereas OSH1 was previously proposed to function also in SAM maintenance, Oskn3 may be involved in patterning organ positions, as its expression was found to mark the boundaries of different embryonic organs following SAM formation. The expression pattern of Oskn2 suggested an additional role in scutellum and epiblast development. Transgenic expression of Oskn2 and Oskn3 in tobacco further supported their involvement in cell fate determination, like previously reported for Knotted1 and OSH1 ectopic expression. Whereas Oskn3 transformants showed the most pronounced phenotypic effects during vegetative development, Oskn2 transformants showed relatively mild alterations in the vegetative phase but a more severely affected flower morphology. The observation that the KNOX genes produce similar though distinct phenotypic reponses in tobacco, indicates that their gene products act on overlapping but different sets of target genes, or that cell-type specific factors determine their precise action.
This study focuses on explaining differences in soft X-ray reflectivity observed for Mo/Si multilayers produced by e-beam evaporation with two different additional methods: ion etching of the Si ...layer and optimization of the substrate temperature during deposition. A 32-period multilayer made with ion etching has a near normal incidence reflectivity of 50%, while the one made at an optimum deposition temperature shows only 41%.
The values for the interface roughness or layer thickness errors obtained from the analysis of small angle reflectivity measurements cannot explain this difference, but a different morphology of the Si and Mo layers could. Extended X-ray absorption fine structure (EXAFS) experiments have been carried out at the SiK and the MoK edge to obtain information on the multilayer structure and chemical composition.
The results of these EXAFS measurements on the two types of multilayers indicate that both the Si and the Mo have an amorphous structure and that no MoSi compounds are formed at the interfaces. However, it is found that multilayers made with ion etching have a reduced fraction of oxygen (≤ 0.5%) in the Si layer compared to layers produced without etching (4.7%). The latter fraction of oxygen, orginating from the background gas in the deposition system, results in higher absorption of soft X-ray radiation and therefore affects the near normal incidence reflectivity. Although both the Si and Mo layers produced with the three different deposition methods are amorphous, some structural differences have been found using the EXAFS analysis.
Intestinal microvascular permeability was studied in the isolated vascularly perfused small intestine of the rat by arterial injection of tracer molecules and collection of venous samples. The ...injection mixture contained a rhodamine-labeled dextran and a fluorescein-labeled dextran or free fluorescein. Pharmacokinetic analysis, based on statistical moment theory, of the tracer outflow concentration-time curve and the application of either the well-stirred model (WSM) or parallel tube model (PTM) was used to assess vasopermeability. The results indicate that the experimental system cannot be considered a pure WSM or a PTM. No different intrinsic clearance (Clint,i) values were found by applying the two models: Clint,i (in ml/min) = 1.23 +/- 0.14 (radius 0.5 nm); 0.44 +/- 0.09 (radius 1.4 nm); 0.31 +/- 0.08 (radius 2.2 nm); 0.02 +/- 0.01 (radius 6.0 nm); and 0 (radius 20.8 nm). Infusion of histamine (10(-5)-10(-3) M) and destruction of the endothelium via perfusion with distilled water increased the permeability for the tracers. We have established a technique for measurement of microvascular permeability characteristics in the rat small intestine. Histamine-induced changes and destruction of the endothelium can be detected in a quantitatively reliable way.
A quantitative procedure to characterize NK cell populations based on the dependence of the frequency of conjugation (alpha) on the effector-to-target ratio (R) is shown. To this end, a detailed ...study of the influence exerted by: (a) the value of R; (b) the number of effector and target cells (N, T); and (c) the source (donor) and enrichment of the effector cell population on the frequency of conjugation between NK effector and K562 target cells has been performed. This has demonstrated that for a given value of R large differences in the values of alpha can be obtained for different donors and/or N values. Hence, the usual practice of reporting the frequency of conjugation at a given value of R cannot be used as a valid criterion for comparison, and this could explain the differences in the alpha values reported in the literature for the same effector-target system. Moreover, the frequency of conjugation depends on the enrichment of the effector cell populations, although it has been shown that in all cases a plot of 1/alpha vs. R for N = constant is always linear with intercept 1/alpha max.alpha max represents the maximum frequency of conjugation for an effector-target system and remains constant for all values of R and N, and is also independent on the donor of the cell source. These characteristics make that the values of alpha max can be used as an easy criterion to determine with accuracy conjugate frequencies in an effector-target system, and could also be applied to characterize the activation or inhibition of effector cell populations by monoclonal antibodies or other agents. This criterion was applied to characterize the enrichment of NK cell populations and so, a value of alpha max = 58 +/- 3% has been obtained when highly purified (greater than or equal to 99%) NK effector cells obtained by panning with the monoclonal antibodies Leu-2, Leu-3 and Leu-4 are used. However, the corresponding value for MDC (14% NK cells) was lowered to 26 +/- 1%.
Det danske sundhedsvæsen og sundhedsfaglige uddannelser er optaget af åndelig-eksistentiel omsorg og trosmæssige livsdimensioner, men samtidigt oplever patienter og pårørende ikke, at deres ...åndelige-eksistentielle behov bliver mødt tilstrækkeligt. Dette skyldes personlige og faglige barrierer hos de sundhedsprofessionelle. Øget praksisnær viden og øvelse på dette område kan afhjælpe disse barrierer. I denne artikel skildrer vi praksisnær religionspsykologisk viden til fagprofessionelle (præster, psykologer, almen praktiserende læger, sundhedsprofessionelle, lærere, etc.), der ønsker at drage omsorg for kriseramte mennesker åndelige og trosmæssige behov. Vi præsenterer modellen »Search for the Sacred« samt flere religionspsykologiske begreber; hellige kerne og ring, helliggørelse, åndelige følelser, åndelige relationer, hellig motivation og hellige øjeblikke. Artiklen diskuterer disse religionspsykologiske perspektiver i forhold til nogle teologiske begreber. Til sidst inddrager vi interviews med troende kriseramte mennesker til at eksemplificere den religionspsykologiske viden, og vi benytter ligeledes disse til at pege på, hvordan den præsenterede viden kan inddrages af fagprofessionelle i deres møde med mennesker i krise og svære livssituationer, med henblik på at imødekomme og understøtte den hjælpsøgendes åndelige og trosmæssige behov ogressourcer.
We have recently described potent antibacterial activity of purified human NK cells. Here we show that this function is regulated by T cytotoxic/suppressor CD8+ cells. Thus, coculture of NK and CD8+ ...cells for 3 h or longer times abrogated the expression of the NK antibacterial activity, and of two activation markers IL-2R and transferrin receptor (Tf-R). The suppressive activity was mediated by PGE2 as demonstrated by direct PGE2 determination in CD8+ cell free supernatants, and by inhibition of CD8+ cell suppression with indomethacin or piroxicam in vitro. We also found that resting T cytotoxic/suppressor cells purified by negative selection produce higher amounts of PGE2 than adherent cells like monocytes and macrophages, and that these concentration levels are in the range of concentrations known to suppress a significant number of in vitro immunologic functions.
A quantitative model for the population distributions of the different types of conjugates formed between cytotoxic T lymphocytes and target cells has been developed. The comparison of the ...theoretical predictions with data of the literature reveals that the transit populations among the different types of conjugates depends on the lymphocyte-to-target ratio, R, and two constants, k and k1. These constants (where k greater than k1) govern, respectively, the transit populations among conjugates of the type LTi (LTn---LTn-1---...LT), and among LjT conjugates (LT---L2T---...---LmT). We have found that high ratios are necessary to obtain conjugates where multiple T lymphocytes are bound to one target cell, and that under these conditions the predominant conjugate, LjT, varies according to j = 1 + k1R. Conversely, for low values of R the predominant population is of the type LTi, where i also shows a linear dependence on R. Our model explains also why the conjugate LT is normally the predominant population under the experimental conditions reported in the literature. A discussion of the influence exerted by the population distributions of lymphocyte-target cell conjugates on the kinetic of the lytic process for these kinds of effector-target systems has also been made.
We have recently shown that purified human NK cells, both resting and activated, efficiently kill gram-negative and gram-positive bacteria in vitro. To investigate the mechanism of NK cell-mediated ...cytotoxicity against Escherichia coli we have developed a mathematical model of the kinetics using the experimental data. The kinetics of killing are characterized by initial target bacterial multiplication, followed by rapid bacterial death. Experiments demonstrates that for each donor there is a threshold number of effector cells necessary to observe a net killing effect. Below the threshold, even use of high effector-to-target ratios lack killing activity and the bacterial growth cannot be stopped. In contrast, if the number of NK cells is larger than the threshold, complete killing is achieved, even at ratios as low as 1/1000. The threshold number varies among donors, ranging between 1200 and 12000 purified NK cells/tube, and provides a quantitative measure of antibacterial activity. Performing the assay at 4 degrees C raised the threshold number required for killing. Experiments performed in Boyden chambers confirm that NK cell-bacteria contact is not necessary for efficient killing, although the kinetics of bacterial lysis is slower. The fit between model and data supports the hypothesis that the bactericidal mechanism is extracellular and is mediated by an anti-microbial factor released from NK cells. Accumulated evidence also indicates that this factor is distinguishable from the mechanisms mediating tumor cell cytotoxicity.
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