Influence of perfusion and cyclic compression on proliferation and differentiation of bone marrow stromal cells in 3-dimensional culture

Until now, there has been no in vitro model that duplicates the environment of bone marrow. The purpose of this study was to analyze proliferation and differentiation of human bone marrow stromal cells (hBMSC) under the influence of continuous perfusion and cyclic mechanical loading. hBMSC of seven individuals were harvested, grown in vitro, and combined. 10(6) hBMSC were seeded on a bovine spongiosa disc and incubated in a bioreactor system. Cell culture was continued using three different conditions: Continuous perfusion (group A), 10% cyclic compression at 0.5Hz (group B) and static controls (group C). After 24h, 1, 2, and 3 weeks, we determined cell proliferation (MTS-assay) and osteogenic differentiation (osteocalcin ELISA, Runx2 mRNA). Tenascin-C mRNA was quantified to exclude fibroblastic differentiation. In groups A and B, proliferation was enhanced after 2 weeks (48.6+/-19.6x10(3) (A) and 44.6+/-14.3 x 10(3) cells (B)) and after 3 weeks (46.6+/-15.1 x 10(3) (A) and 44.8+/-10.2 x 10(3) cells (B)) compared with controls (26.3+/-10.8 x 10(3) (2 weeks) and 17.1+/-6.5 x 10(3) cells (3 weeks), p<0.03). Runx2 mRNA was upregulated in both stimulated groups after 1, 2, and 3 weeks compared to control (group A, 1 week: 5.2+/-0.7-fold; p<0.01, 2 weeks: 4.4+/-1.9-fold; p<0.01, 3 weeks: 3.8+/-1.7-fold; p=0.013; group B, 1 week: 3.6+/-1.1-fold, p<0.01, 2 weeks: 4.2+/-2.2-fold, p<0.01; 3 weeks: 5.3+/-2.7-fold, p<0.01). hBMSC stimulated by cyclic compression expressed the highest amount of osteocalcin at all time points (1 week: 294.5+/-88.4 mg/g protein, 2 weeks: 294.4+/-73.3mg/g protein, 3 weeks: 293.1+/-83.6 mg/g protein, p0.03). The main stimulus for cell proliferation in a 3-dimensional culture of hBMSC is continuous perfusion whereas mechanical stimulation fosters osteogenic commitment of hBMSC. This study thereby contributes to the understanding of physical stimuli that influence hBMSC in a 3-dimensional cell culture system.     

The low-molecular-mass subunit of the cell wall channel of the Gram-positive Corynebacterium glutamicum. Immunological localization, cloning and sequencing of its gene porA.

The 5-kDa protein PorA of the Gram-positive bacterium Corynebacterium glutamicum is the subunit of the cell wall channel. Antibodies raised against PorA specifically detected the protein on the cell surface. PorA was sequenced using Edman degradation and a gas phase sequencer. The primary sequence was used to create degenerate oligonucleotide primers. The gene of the channel-forming protein and its flanking regions were obtained by PCR followed by inverse PCR. The gene porA comprises 138 bp and encodes a 45-amino-acid-long acidic polypeptide with an excess of four negatively charged amino acids in agreement with the high cation selectivity of the PorA cell wall channel. PorA does not contain an N-terminal extension. A ribosomal-binding site was recognized 6 bp before the start codon ATG of porA. It codes for the smallest subunit of a membrane channel known so far and for the first cell wall channel protein of a corynebacterium. Southern blots demonstrated that only the chromosomes of corynebacteria contain homologous sequences to porA; no hybridization could be detected with DNA from other mycolata.     

Particle size distribution in gypsic soils

Summary An investigation has been made of the effects upon soil dispersion of different sized particles of gypsum and of methods of counteracting this effect. A final, recommended procedure for particle size determination in gypsic soils has been formulated and which depends upon the fact that treatment of gypsum crystals with barium chloride solution forms a protective coating of barium sulphate which isolates the gypsum from all further reaction in an alkaline medium. Satisfactory results have been obtained for soils containing up to over 90 percent gypsum, even if present in very fine particles of less than 0.05 mm.Food & Agriculture Organization of the United Nations     

Die postembryonale pterylose bei taubenrassen verschiedener grösse

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In Vivo Labeling by CD73 Marks Multipotent Stromal Cells and Highlights Endothelial Heterogeneity in the Bone Marrow Niche

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Karyologische Anatomie von Zoocecidien und ihre Kernstrukturen

Zusammenfassung Von den 60 untersuchten Cecidien erbrachten bezüglich des Vorkommens somatischer Polyploidie 26 positive, 9 vermutlich negative und 25 eindeutig negative Ergebnisse. In 25 Gallen wurden endopolyploide Kerne mit Hilfe der Analyse der Kernstruktur und der statistischen Erfassung der Kernvolumina nachgewiesen. Es wurden folgende höchste Endopolyploidiestufen in den Nährgeweben der zwölf näher untersuchten endopolyploiden Gallen festgestellt (in Klammern die höchsten in den übrigen Gallengeweben beobachteten Polyploidiegrade): beiNeuroterus numismalis 512n (16n), beiAndricus marginalis 1024n (16n), beiNeuroterus quercusbaccarum 512n (8n), beiAndricus ostrea 32n (4n), beiAndricus aries 512n (4n), alle bisher genannten aufQuercus robur; beiCynips agama aufQuercus petraea 128n (16n), beiEriophyes similis pruni spinosae aufPrunus spinosa 512n (4n), beiAceria tristriata aufJuglansregia 256n (16n), beiAceria eucriotes aufLycium barbarum 128n (256n in Epidermiszellen), beiSierraphytoptus setiger aufFragaria viridis 32n (2n), beiJanetiella lemei aufUlmus minor 128n (16n); die Kerne der Epidermiszellen der Gallen vonDasyneura affinis aufViola odorata werden bis zu 256ploid (im unbeeinflußten Blatt 16n), die Haare an der Außente werden bis 256ploid (die Haare des unbeeinflußten Blattes bis zu 32ploid), dagegen sind die stark aufgelockerten Kerne der Nährzellen dieser Galle wahrscheinlich bloß tetraploid. Mit den Werten 512n bzw. 1024n wurden die bisher höchsten außerhalb des Blütenbereiches auftretenden Endopolyploidiegrade bei Angiospermen gefunden.Auch in manchen anderen, nicht näher untersuchten Cecidien — meist Cynipidengallen, aber auch Acaro- und Dipterocecidien — treten endomitotisch vergrößerte Kerne auf. In allen untersuchten Gallen von Tenthrediniden (Blattwespen), Aphididen (Blattläusen), in vielen Acarosowie in manchen Dipterocecidien, weiters in allen Fällen, in denen Homopteren Pflanzen befallen, ohne eigentliche Gallen zu bilden, in allen Bakterien-, aber auch in einer Cynipidengalle fanden sich keine endopolyploiden Kerne.Der Grad der somatischen Polyploidie steigt meistens mit der Annäherung an den Parasiten an; eine Ausnahme bilden die beiden untersuchten Dipterengallen, bei denen sich die höchsten Polyploidiestufen nicht in unmittelbarer Umgebung der Larve, sondern weiter außen finden.Die Endochromosomen der endopolyploiden Kerne mancher Gallen sind zu Bündeln vereinigt (beiNeuroterus numismalis ,Andricus marginalis undDasyneura affinis), in anderen kommt es nur zur Bil dung verschieden gebauter Endochromozentren (beiAceria tristriata, Eriophyes similis pruni spinosae undJanetiella lemei); es treten aber auch, in ein und demselben Kern neben Endochromozentren auch Bündel von Endochromosomen auf (beiAndricus aries undNeuroterus quercusbaccarum ). Zumeist ergaben sich zwischen den gleichen Gallen, die von verschiedenen Pflanzenindividuen derselben Art stammten, keine greifbaren Unterschiede; nur bei denen, die von einem Exemplar vonQuercus robur f.fastigiata gesammelt wurden, trat ein Dimorphismus in der Ausbildung der dem Nukleolus assoziierten Bündel von Endochromosomen auf. In den hoch endopolyploiden Kernen der Gallen vonAndricus aries aufQu. robur f.fastigiata finden sich in den Bündeln der Endochromosomen gewisse Anklänge an den Scheibenbau der Dipteren-Riesenchromosomen.

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Summary It has been proved that endopolyploid nuclei exist in many of the investigated plant galls. They are to be found typically in most galls produced by gall-flies (Cynipidae), and also frequently in galls produced by mites (Acarina — Tetrapodili) or Diptera — but not in galls produced by saw-flies (Tenthredinidae) and Homoptera. Endopolyploid nuclei occur mostly in nutritive tissues but frequently in other tissues also; in the nutritive tissue ofAndricus marginalia there are to be found nuclei up to 1024n: this is the highest degree of endopolyploidy known from parts outside the angiosperm floral region. The degree of endopolyploidy increases commonly from the periphery of the gall towards the parasite. The endopolyploid nuclei contain either endochromosomes retained in bundles or variously formed endochromocentres; in some cases they contain both.

     

Role of Cyclic Adenosine 3′,5′-Monophosphate in the In Vivo Expression of the Galactose Operon of Escherichia coli

Studies of levels of galactokinase in Escherichia coli with mutations affecting synthesis of, or response to, cyclic adenosine 3',5'-monophosphate show that this nucleotide does not play a major role in expression of the galactose operon, causing at most a twofold stimulation. The discrepancy between our in vivo results and the marked stimulation by cyclic adenosine 3',5'-monophosphate in in vitro systems indicates that current cell-free systems lack a factor which allows efficient expression of the galactose operon even in the absence of cyclic adenosine 3',5'-monophosphate or of the binding protein for this nucleotide.     

Referate

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Coccystes undoxylophus

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