Pathways of lymphocyte migration within the periarterial lymphoid sheath of rat spleen

Summary Male Wistar rats were injected intravenously with 5-(³H)uridine-labeled lymphocytes isolated from lymph nodes of syngeneic donors and enriched in T cells. After short periods of time (3 to 120 min after injection), labeled lymphocytes were localized in spleen compartments using autoradiography to identify routes of lymphocyte movement from blood into splenic parenchyma and to follow migration pathways of recirculating lymphocytes within the periarterial lymphoid sheath (PALS). Topographical analysis of labeled lymphocytes was performed in specific planes of PALS characterized by the diameter of the arterial vessel and termed PALS large, PALS medium, and PALS small (PALS L, PALS M, PALS S, respectively). Attention was also paid to accumulations of labeled lymphocytes close to the arterial vessel wall. Initially, labeled lymphocytes were localized in PALS S and PALS M near the terminal branching of arterial vessels and in the marginal zone (MZ). We conclude that lymphocytes emigrate from blood into splenic parenchyma within two white pulp compartments: in MZ, and directly within PALS through the wall of capillary vessels. The sequential accumulation of labeled cells near arterial vessels of increasing diameter suggests that the recirculating pool of lymphocytes migrates into the central part of PALS L by two routes: from MZ, and along arterial vessels from PALS S and PALS M.R.B. was a fellow of the Alexander von Humboldt-Stiftung, on leave from the Department of Histology and Embryology, Institut of Biostructure, Academy of Medicine, ul. Swiecickiego 6, PL-60-781 Pozna, Poland.     

Polarity and competition between megaspores in the ovule ofOenothera hybrids

New data on the development of polarity in the ovules during megasporogenesis and early stages of embryo sac development inOenothera-hybrids are presented. It is confirmed that allOe. hookeri-hybrids show a strong tendency to form heteropolar tetrads, with the micropylar megaspore developing into an embryo sac. This preference is seen in the delay of the second meiotic division on the chalazal side, the absence of callose in the lateral wall of the micropylar megaspore, and the accumulation of starch in this megaspore. However, homopolar tetrads, chalazal preference, and ovules with two developing embryo sacs are also observed with considerable frequency. Quantitative data on the frequency of the different developmental types are compared with earlier genetic results about competition in the haplophase. There is sufficiently good agreement to support the hypothesis ofRenner that there is a correlation between the developmental processes in the megaspore tetrad and the genetic phenomena of competition in the haplophase.     

Mouse X chromosome

Overall, the probe map fromDXWas70 toAmg encompasses 72 cM and includes 103 loci. Eight of these have been designated reference loci (see Table 2 and previous section) on account of their wide usage that would enable the cross reference of independent maps created in different laboratories. Reference loci are to be readily available and easily used probes for Southern hybridization. By and large, they will be STSs, regeneratable by PCR, and correspond to a known gene. In addition, on the mouse X Chr, there is a reference locus from each of the known conserved linkage groups found between the mouse and human X Chrs. All the loci, barDXWas31, represent conserved sequences. Committee Members: D. Adler, P.J. Barnard, Y. Boyd, N. Brockdorff, J. Derry, C. Disteche, C. Faust, M.F. Lyon, S. Rastan, M. Seldin and L. Siracusa.     

Cellulolytic enzymes for the hydrolysis of leached beet cosette

Summary Cellulolytic fungi were isolated from rotting leaves and tested for extra-cellular cellulase activities (CMCase, avicelase, cellobiase and xylanase). The effect of the proportion of the enzyme activities on the rate of degradation of leached beet cosette was observed using a range of supernatant fluids in appropriate combinations. At low cosette concentrations (1.5–3.0 g/l), avicelase and cellobiase were the rate limiting enzymes; avicelase in the initial stages of reaction and cellobiase after 6–8 hours, when cellobiose inhibition becomes important. A ratio of celiobiase to avicelase of approx 2.0 was established as appropriate. At higher substrate concentrations (10 g/l, 40 g/l) the best cellobiase to avicelase ratio was maintained and up to 40% hydrolysis was obtained in the 10 g/l incubation with 10 Uav/l and 20 Ucellob/l. At 100 g/l cosette concentration, substrate inhibition was observed.     

Preparation of an enantiomerically pure, highly porous metalloorganic crystal

We report the synthesis and the structure determination of tris-(4,5-diazo-spiro-bifluorene)ruthenium(II) chloride, a chiral building block whose racemic mixture solution spontaneously resolves and forms two crystalline, enantiomerically pure, porous networks composed exclusively of the Λ or Δ atropisomers. The extended chiral channels are occupied by water molecules (approximately 20% by weight) and the chloride counter-ions.     

CHRONOCRISIS: When Cell Cycle Asynchrony Generates DNA Damage in Polyploid Cells

Polyploid cells contain multiple copies of all chromosomes. Polyploidization can be developmentally programmed to sustain tissue barrier function or to increase metabolic potential and cell size. Programmed polyploidy is normally associated with terminal differentiation and poor proliferation capacity. Conversely, non-programmed polyploidy can give rise to cells that retain the ability to proliferate. This can fuel rapid genome rearrangements and lead to diseases like cancer. Here, the mechanisms that generate polyploidy are reviewed and the possible challenges upon polyploid cell division are discussed. The discussion is framed around a recent study showing that asynchronous cell cycle progression (an event that is named “chronocrisis”) of different nuclei from a polyploid cell can generate DNA damage at mitotic entry. The potential mechanisms explaining how mitosis in non-programmed polyploid cells can generate abnormal karyotypes and genetic instability are highlighted.     

Targeted Exosomes for Drug Delivery: Biomanufacturing,Surface Tagging,and Validation

Exosomes hold great potential to deliver therapeutic reagents for cancer treatment due to its inherent low antigenicity. However, several technical barriers, such as low productivity and ineffective cancer targeting, need to be overcome before wide clinical applications. The present study aims at creating a new biomanufacturing platform of cancer‐targeted exosomes for drug delivery. Specifically, a scalable, robust, high‐yield, cell line based exosome production process is created in a stirred‐tank bioreactor, and an efficient surface tagging technique is developed to generate monoclonal antibody (mAb)‐exosomes. The in vitro characterization using transmission electron microscopy, NanoSight, and western blotting confirm the high quality of exosomes. Flow cytometry and confocal laser scanning microscopy demonstrate that mAb‐exosomes have strong surface binding to cancer cells. Furthermore, to validate the targeted drug delivery efficiency, romidepsin, a histone deacetylase inhibitor, is loaded into mAb‐exosomes. The in vitro anti‐cancer toxicity study shows high cytotoxicity of mAb‐exosome‐romidepsin to cancer cells. Finally, the in vivo study using tumor xenograft animal model validates the cancer targeting specificity, anti‐cancer efficacy, and drug delivery capability of the targeted exosomes. In summary, new techniques enabling targeted exosomes for drug delivery are developed to support large‐scale animal studies and to facilitate the translation from research to clinics.