Immunocytochemical location of vitellin in the egg of the silkworm,Bombyx mori,during early developmental stages

Summary Vitellin was purified from eggs of the silkworm,Bombyx mori, by a new method in which vitellin was extracted from isolated yolk granules. The purified vitellin had a molecular weight of 540,000. An antibody against purified vitellin was prepared in rabbits. It reacted with the hemolymph vitellogenin as well as with purified vitellin, but not with other proteins in the hemolymph or in the extract from yolk granules. The anti-vitellin IgG was used to immunocytochemically locate vitellin in theBombyx non-diapause egg during early developmental stages. In the egg, just after oviposition, vitellin was located in internal yolk granules and in small yolk granules of the periplasm. During the early developmental stages studied, vitellin was not metabolized uniformly throughout the egg. The vitellin of the internal yolk granules located at the posterior-dorsal part and of the small peripheral yolk granules was utilized in 16 h and 2 days, respectively, after oviposition. A thin, very vitellin-poor layer was located between the periplasm and the vitellin-rich interior in the newly laid egg. it was always in close contact with the periphery where blastoderm and germ-band cells developed.     

Further characterization of the aggregation and fusion of chromaffin granules by synexin as a model for compound exocytosis

Synexin was isolated from bovine liver and found to aggregate adrenal chromaffin granules in the same Ca2+-dependent manner as previously described for adrenal synexin. The chromaffin granule aggregating activity of liver synexin was blocked in vitro by the addition of an antibody prepared to the 47,000 molecular weight band extracted from an SDS gel of an adrenal medullary synexin preparation. Chromaffin granules aggregated by synexin fused when exposed to cis-unsaturated fatty acids at concentrations comparable to those released from phospholipids by stimulated secretory cells. The synexin-induced aggregation reaction was blocked by Erythrosin B, a common food coloring, and by the phenothiazine antipsychotic trifluoperazine and promethazine. The aggregation and fusion of chromaffin granules thus appears to be a useful model system for studying synexin from diverse tissues and for testing pharmacologically or toxicologically active substances for effects on secretory systems.     

Isolation and characterization of noradrenalin storage granules of bovine adrenal medulla

A method is described for the preparation of (1) the heavy population of bovine adrenal chromaffin granules ($\text{S}{}_{\mathrm{<mtext>H</mtext>}}\text{(}\text{average sedimentation coefficient}\text{) = 12 400}\text{S}$ in 0.25 M sucrose) essentially free from contamination with mitochondria and other organelles, and (2) a subpopulation of this heavy population which is highly enriched in noradrenalin ($\text{?95\%}$ of the total catecholamine is noradrenalin). The method is based on isopycnic gradient centrifugation using a self-generating gradient of polyvinylpyrrolidone-coated colloidal silica particles (Percoll) in 0.5 M sucrose medium.The isolated population of noradrenalin granules appeared highly electron dense in transmission electron microscopy and revealed a rather narrow size distribution. The specific content of amine and adenine nucleotides (with reference to total granule protein) was markedly higher than for the total population of heavy chromaffin granules. The molar ratio of amines to adenine nucleotides was, however, lower in the noradrenalin granules, i.e. 4.8 vs. 11.9.     

Ultrastructure of retrocerebral complex of the earwig, Euborellia annulipes, and rôle of aorta as a neurohaemal organ

The ultrastructure of the retrocerebral endocrine-aortal complex of the earwig, Euborellia annulipes has been studied. The space between the inner and outer stromal layers of the aorta is occupied by numerous axon terminals and pre-terminals containing large electron dense granules (NS-I) of approximately 100 to 220 nm and a few axon terminals having small granules (NS-II) of approximately 40 to 90 nm; the former appear to belong to medial neurosecretory A-cells, and the latter to the B-cells of the brain. The corpora cardiaca consist of intrinsic cells with mitochondria and multivesicular bodies. Granules of type NS-II and NS-III are observed in the axon terminals and pre-terminals in the corpora cardiaca. The NS-II are identical to those found in the aorta and are probably the secretions of the lateral B-cells. Granules of type NS-III are 40 to 120 nm and electron dense, and are intrinsic in origin. Similar granules occur in the intrinsic cells of the corpora cardiaca. E M studies have confirmed the rôle of the aorta as a neurohaemal organ for the medial neurosecretory cells, and the corpora cardiaca for the lateral neurosecretory cells of the brain. The corpora cardiaca also act as a reservoir for the intrinsic secretion. The corpus allatum is a solid body consisting of parenchymal cells with prominent nuclei, mitochondria, and endoplasmic reticulum. In between its cells are occasional glial cells and also neurosecretory as well as non-neurosecretory axons. The gland is devoid of A-cell NSM.     

Polyphosphate granules in encysted zoospores of Rozella allomycis

Methods of ultracytochemistry and of X-ray energy dispersive analysis have been used to demonstrate that the gamma-like granules in encysted zoospores of the chytrid Rozella allomycis contain polyphosphate. The possibility that cysts contain two classes of polyphosphate granules which differ in structure, in function, and in origin is discussed.     

An inhibitor specific for the mouse T-cell associated serine proteinase 1 (TSP-1) inhibits the cytolytic potential of cytoplasmic granules but not of intact cytolytic T cells

We have investigated a proteinase inhibitor, designed according to the preferred amino acid sequence that is cleaved by the murine T-cell specific serine proteinase 1 (TSP-1) for its effect on the cytolytic potential of cloned cytotoxic T-cell lines (CTLL) and of cytoplasmic granules, derived from these cells. Pretreatment of effector cells with H-D-Pro-Phe-Arg-chloromethyl-ketone (PFR-CK) prior to the cytotoxicity assay did not result in inhibition of cytolytic activity of three independent CTLL and did not effect their granule-associated TSP-1 activity after extraction with Triton X-100. Furthermore, PFR-CK did not interfere with cytolysis of target cells by CTLL when present for the entire incubation period. In contrast, PFR-CK inhibited in a dose-dependent manner both TSP-1 activity and the hemolytic/cytolytic potential of isolated cytoplasmic granules after their pretreatment with high-salt concentration. We interpret these results to mean that cytolysis of target cells by CTLL involves the granule-associated proteinase TSP-1, which probably becomes active upon exocytosis following effector-target cell interactions.     

NOTE—THE STA-1 MUTATION PREVENTS ASSEMBLY OF STARCH GRANULES IN NITROGEN-STARVED CELLS AND SERVES AS A USEFUL MORPHOLOGICAL MARKER DURING SEXUAL REPRODUCTION IN CHLAMYDOMONAS MONOICA (CHLOROPHYCEAE)

Iodine staining of clones of nitrogen-starved Chlamydomonas cells was used to screen for mutants with altered levels or altered composition of storage starch. Mutations leading to defects in quantity or morphology of starch granules not only can provide information on storage starch biosynthesis and granule assembly but can also be used as morphological markers in genetic and cell biological studies. A mutant of Chlamydomonas monoica Strehlow devoid of starch granules was obtained following ultraviolet mutagenesis. Nitrogen-starved cells of the sta-1 strain lacked pyrenoidal starch granules and granules normally associated with thylakoid membranes. The mutant phenotype was the consequence of a single Mendelian mutation that appeared to affect granule assembly rather than starch biosynthesis per se and that had no effect on vegetative growth, sexual reproduction, or zygospore viability.     

The soluble components of chromaffin granules a carbon-13 NMR survey

Carbon-13 NMR spectra of the reconcentrated chromaffin granule lysate have been obtained at 50 MHz and 62.9 MHz. The spectrum contains a number of assignable resonances in addition to those of the main soluble components (catecholamines, adenine nucleotides and chromogranin). Guanine and uridine nucleotides are present at levels of 0.13 and 0.08 mol/mol adenine nucleotides, respectively. Concentrations of cytidine nucleotides and NAD⁺ are below the detection limit (0.02 mol/mol adenine nucleotides). An unidentified low molecular weight species, thought to be an adenine-containing oligonucleotide, is also present. Ascorbic acid was observed at a concentration of 0.14 mol/mol adenine nucleotides, but both dopamine and dehydroascorbic acid were below the detection limit. Protein resonances agree well with the reported amino acid composition of chromogranin A, with the exception of tryptophan and glutamine which have not previously been measured. The concentrations of these residues are estimated to be 12 ± 3 and 39 ± 5 residues per 77 000 dalton unit of chromogranin A. Substantial intensity due to unsaturated fatty acid side-chains in solubilized lipid is seen in the olefinic carbon region and in the methylene region, suggesting the presence of lipoprotein. Unassigned carbohydrate resonances are also present, but are largely obscured by sucrose in the isolation medium.     

解毒颗粒联合阿帕替尼治疗中晚期肝癌的回顾性研究

目的:评估解毒颗粒联合阿帕替尼治疗中晚期肝癌患者的疗效及其不良反应。方法:对2018年12月至2019年6月收治于海军军医大学第一附属医院口服解毒颗粒联合阿帕替尼的27例肝癌患者的临床资料进行回顾性研究。无法切除或复发的中晚期肝癌患者被纳入研究,给予解毒颗粒联合阿帕替尼治疗直至疾病进展或不可耐受其毒副反应,随访观察治疗效果、生存期、炎症因子指标及不良反应。结果:治疗后完全缓解(CR)4例(14.81%),部分缓解(PR)4例(14.81%),稳定(SD)8例(29.63%),进展(PD)11名患者(40.74%),疾病控制率(DCR)为59.26%(16/27),客观缓解率(ORR)为29.63%(8/27)。中位无进展生存期(PFS)为3.630个月,中位总生存期(OS)为13.667个月。常见的不良反应是高血压59.26%(16/27)、蛋白尿59.26%(16/27)、腹泻74.07%(20/27)以及手足综合征62.96%(17/27)。治疗后炎症因子指标中C反应蛋白、白介素2水平下降,存在统计学差异(P0.05)。结论:解毒颗粒联合阿帕替尼治疗中晚期肝癌安全、有效,可降低患者炎症反应,不良反应可耐受。