Freeze-Fracture Study of the Bloodstream Form of Trypanosoma brucei gambiense

The ultrastructure of Trypanosoma brucei gambiense was investigated by the freeze-fracture method. Three different regions of the continuous plasma membrane; cell body proper, flagellar pocket, and flagellum were compared in density and distribution of the intramembranous particles (IMP's). The IMP-density was highest in the flagellar pocket membrane and lowest in flagellum. Intra membranous particles of the cell body membrane were distributed uniformly on both the protoplasmic (P) and exoplasmic (E) faces. On the P face of the flagellar membrane, a single row of IMP-clusters was seen along the juncture of the flagllum to the cell body. Since the spacing of the IMP-clusters was almost equal to the spacing of the paired rivet structures observed in thin section, these clusters likely are related to the junction of flagellum and cell body. At the neck of the flagellar pocket, several linear arrays of IMP's were found on the P face of the flagellar membrane, while on the E face rows of depressions were seen. At the flagellar base, the clusters of IMP's were only seen on the P face. On the flagellar pocket membrane, particle-rich depressions and linear particle arrays were also found on the P face, while on the E face such special particle arrangements were not recognized. These particle-rich depressions may correspond to the sites of pinocytosis of the bloodstream forms which have been demonstrated in thin sections.     

Aspects Regarding the Action of CCC in Hollyhock Plants

CCC [(2-chloroethyl) tnmethyl ammonium chloride] retarded shootelongation and increased leaf thickness and size (or volume)of leaf mesophyll cells in hollyhock plants. Photosyntheticactivities were stimulated by CCC, whereas post-illuminationCO₂ outburst-1 [PIB-1], PIB-2 and dark respiration were depressedby CCC. Chlorophyll and protein content of leaves treated withCCC also increased as compared with those of control leaves.CCC enabled the plant to bear flowers at the nodes from lowerto upper positions of the hollyhock shoots without flower (bud)abortion. Key words: Photosynthesis, CCC, mesophyll cell     

Freeze-Fracture Localization of Filipin-Sterol Complexes in Plasma- and Cyto-Membranes of Pneumocystis carinii1

The polyene antibiotic, filipin, was used as the probe for demonstrating sterols in the freeze-fractured plasma- and cytomembranes of Pneumocystis carinii. The distribution of filipin-sterol complexes was homogeneous on the plasma membrane throughout all developmental stages from trophozoite to cyst; however, the density of the complexes gradually decreased with the progress of development. In the trophozoite, the density of the complexes was 485 ± 42/μm² on the P face and 341 ± 27/μm² on the E face. It was 249 ± 50 on the P face and 132 ± 48 on the E face in the precyst and 138 ± 24 and 59 ± 20, respectively, in the cyst. The membranes of nucleus, mitochondria, and small round bodies showed more or fewer complexes while no complexes were found in the membranes of one endoplasmic reticulum. In nuclear and mitochondrial membranes, some small scattered clusters of complexes were observed. Two types of vacuoles were distinguished: one having many complexes in its membrane and the other having none at all.     

Analysis of Cellular Mosaicism in a Transgenic Mouse by Histological In Situ Hybridization

One of the transgenic mice carrying a chicken δ-crystallin gene was found to be mosaic with regard to the distribution of the exogenous gene. Taking advantage of the exogenous DNA sequences as a cell lineage marker detectable by histological in situ hybridization technique, we studied cellular mosaicism in mouse 7–5. This mouse carried the exogenous gene in 20–40% of its cells, probably reflecting chromosomal integration of the exogenous DNA which occurred in a blastomere of around the 4-cell stage. The cells carrying the gene contributed to virtually any kind of tissue and their distribution varied from one tissue to another. For instance, in the neural retina, gene-positive cells formed columns several cells wide, indicating that migration of the cells derived from the founder cells is mainly along the radial axis. However, in other tissues we examined, clusters of the marked cells were less obvious, indicating the occurrence of extensive cell mixing during histogenesis. Thus, mosaic analysis of cell lineage in mouse ontogeny appears meaningful in early developmental stages or when clonal outgrowth takes place in a tissue.     

Sporogony in Pneumocystis carinii: Synaptonemal Complexes and Meiotic Nuclear Divisions Observed in Precysts1

Evidence for meiosis was demonstrated electron microscopically for the first time in Pneumocystis carinii in rat alveoli by the observation of synaptonemal complexes followed by nuclear divisions. Synaptonemal complexes indicating meiotic nuclear divisions were observed in uninuclear precysts. Additionally, owing to the use of tannic acid as a fixative, spindle microtubules were also observed for the first time in the precyst. Based on these facts, a new life cycle of the organism is proposed. The precyst has generally been considered an intermediate form between the trophozoite and the cyst. The present paper proposes that the precyst is additionally defined as the cell in which eight intracystic bodies are produced through meiotic reduction. The most characteristic feature of the precyst is a clump of mitochondria in the cytoplasm. We divide the precyst phase into three forms, which are named early, intermediate, and late. Synaptonemal complexes were only observed in the early precyst, which is a uninuclear cell with a thin pellicle. In the intermediate precyst, nuclear divisions are observed as follows: meiosis I produces two haploid nuclei and each of these divides at meiosis II producing four nuclei. After that, another postmeiotic mitosis takes place, resulting in eight haploid nuclei. In the late precyst, a delimiting membrane originates from the mother plasmalemma and surrounds the daughter nuclei and a small portion of the adjacent cytoplasm. Finally, when the eight intracystic bodies are complete, the precyst changes to a cyst. Thus, we deduce that intracystic bodies resulting from meiotic nuclear division are haploid and, after excystation, they are haploid trophozoites. We consider that this process can be called sporogony. Although we could not distinguish between the haploid and the diploid trophozoite, it is quite plausible that copulation occurs, probably in host alveoli.     

The Relationship of Capsular-type of Staphylococcus epidermidis to Virulence and Induction of Resistance in the Mouse

Biochemical, immunological and morphological properties of mouse virulent Staphylococcus epidermidis strains ATCC-31432, SE-360 and SE-10 isolated from clinical specimens were compared. Heat-killed organisms and cell surface polysaccharides extracted from cell surface fractions induced resistance in mice only against challenge with the homologous strain. Hyperimmune rabbit serum prepared with these strains passively protected mice against challenge infection only with the homologous strain. Protective activity in immune sera was absorbed by homologous whole cell and cell surface polysaccharide, but not by heterologous organisms and cell surface polysaccharide. In agar diffusion tests, cell surface polysaccharides from strains ATCC-31432, SE-360 and SE-10 produced single precipitin lines only with the homologous antiserum. The outermost layer of ultra-thin sections of the three strains was labelled by homologous but not by heterologous ferritin-conjugated serum. Biochemical analysis of the cell surface polysaccharides showed that they were composed of hexosamine, glycerol, phosphorous, alanine, glycine and phenylalanine. The three strains ATCC-31432, SE-360 and SE-10 were regarded as different from each other. Thirteen of 300 fresh isolates of Staph. epidermidis randomly selected from human clinical specimens proved to be virulent for mice. With ATCC-31432, SE-360 and SE-10 tentatively designated as capsular-type I, type II and III, respectively, a majority of mouse virulent strains belonged to capsular-type II.