Limitations in the quantitative separation of DNA samples by density gradient centrifugation

Résumé Des mélanges de l'ADN («léger») et de l'ADN-¹⁴ C-BU («lourd») ont étés centrifugés dans le chloride de caesium pendant 65 h à 60,000g. Le résidu de l'ADN «lourd» dans l'ADN «léger» fut de l'ordre de 0.1 à 1%. La grandeur du résidu n'a pas correspondu à la proportion de l'ADN «lourd», ni à la concentration totale de l'ADN dans le chloride de caesium.     

Isolation of plasmid DNA fromAgrobacterium by isopycnic density gradient centrifugation in vertical rotors

Summary Fixed angle and vertical rotors are compared in terms of their sample load, run time, and resolution for the preparation of plasmid DNA fromAgrobacterium.Acknowledgments. This work was supported by a CASE Studentship from the SRC and ICI Corporate Laboratories to J.D., and a grant from the SRC to M.R.D. and E.C.C. The authors thank Mr B.V. Case for photographic assistance.     

Enrichment in spontaneous and complement dependent rosette forming cell populations using ficoll-hypaque gradient centrifugation

Summary Peripheral blood human T (TL) and B (BL) lymphocytes were separated by continuous Ficoll-Hypaque (FH) gradients. BL were found at the 1050 density interfase (70.5% EAC rosettes) with no TL (0% EAC rosettes); while at the 1068 density interfase there was an enrichment of TL (68% E rosettes) with very few BL (8.5% EAC rosettes).     

Density gradient centrifugation of tumor cells from needle biopsies and their respective source tumors: a comparison of density distributions

Summary A needle biopsy technique was applied to a murine fibrosarcoma (FSa) tumor system. FSa tumors of 8 mm diameter in size were biopsied and then made into single cell suspensions. Resulting density profiles of cells from both sources were compared following centrifugation in Renografin gradients. In all cases, there was excellent agreement between the density profiles of the material from each of the biopsies and the corresponding solid tumors.This investigation was supported by grant CA-18628, awarded by the National Cancer Institute, Department of Health and Human Services.     

Density gradient centrifugation of tumor cells from needle biopsies and their respective source tumors: a comparison of density distributions

A needle biopsy technique was applied to a murine fibrosarcoma (FSa) tumor system. FSa tumors of 8 mm diameter in size were biopsied and then made into single cell suspensions. Resulting density profiles of cells from both sources were compared following centrifugation in Renografin gradients. In all cases, there was excellent agreement between the density profiles of the material from each of the biopsies and the corresponding solid tumors.     

Density gradient centrifugation of cells separated from multicellular tumor spheroids

Summary Cells from a murine fibrosarcoma (FSa) have been grown in vitro as multicell tumor spheroids (MTS). The growth rate of these MTS was determined. Follwing selected periods of growth, MTS were made into a single cell suspension and separated on linear density gradients of Renografin. While only 1 population of cells was separated from small spheroids (400 m diameter), at least 3 subpopulations of tumor cells were separated and isolated from large spheroids (800 m in diameter).This investigation was supported in part by grants numbered CA-18628 and CA-06294, awarded by the National Cancer Institute, DHEW.     

Density gradient centrifugation of cells separated from multicellular tumor spheroids.

Cells from a murine fibrosarcome (FSa) have been grown in vitro as multicell tumor spheroids (MTS). The growth rate of these MTS was determined. Following selected periods of growth, MTS were made into a single cell suspension and separated on linear density gradients of Renografin. While only 1 population of cells were separated from small spheriods (400 mum diameter), at least 3 subpopulations of tumor cells were separated and isolated from large spheroids (800 mum in diameter).     

Production of Friend virus particles at the surface of cells synchronized by Ficoll gradient centrifugation]

Murine cells continuously producing Friend viral complex were synchronized by Ficoll gradient centrifugation. Cells were isolated from different parts of the gradient and counts of budding particles were performed on ultra-thin sections. No significant differences were noticed, suggesting that viral release is not related to cell cycle.     

A new plot to estimate protein molecular weight by density gradient ultracentrifugation

Summary A plot of the logarithm of the molecular weight against the logarithm of the sedimenting distance is proposed for estimation of protein molecular weight. The proteins are separated in acrylamide-containing linear density gradients, polymerized and stained after centrifugation.     

Closed circulation in the rat spleen as evidenced by scanning electron microscopy of vascular casts

Zusammenfassung Rasterelektronenmikroskopische Beobachtungen der Methakrylatausgüsse der Blutgefässe von Rattenmilz ergeben, dass jedes Arteriolenende unmittelbar mit einem Sinus verknüpft ist. Man bekommt nirgends ein Bild, das eine offene Endigung der Arteriolen in die Milzsträge andeutet. Dieses Ergebnis stützt die «geschlossene» Theorie der Milzgefässe, die neuerdings von der «offenen» Theorie überschattet scheint.     

Heterogeneity of gangliosides among T cell subsets

Gangliosides are major components of highly organized membrane microdomains or rafts, yet little is known about the role of gangliosides in raft organization. This is also the case of gangliosides in TCR-mediated activation. Comprehensive structural analysis of gangliosides in the primary thymocytes and CD4⁺ T and CD8⁺ T cells was not achieved due to technical difficulties. We have found that CD8⁺ T cells express very high levels of o-series gangliosides, but on the other hand, CD4⁺ T cells preferably express a-series gangliosides. In the TCR-dependent activation, CD4⁺ T cells selectively require a-series gangliosides, but CD8⁺ T cells do require only o-series gangliosides but not a-series gangliosides. Ganglioside GM3 synthase-deficient mice lacking a-series gangliosides neither exhibited the TCR-dependent activation of CD4⁺ T nor developed ovalbumin-induced allergic airway inflammation. These findings imply that the distinct expression pattern of ganglioside species in CD4⁺ and CD8⁺ T cells define the immune function of each T cell subset.