Simian immunodeficiency virus infection and flow cytometric characterization of Japanese macaque (Macaca fuscata) hematopoietic cells

We characterized Japanese macaque (Macaca fuscata) hematopoietic cells using flow cytometry and identified 28 cross-reactive anti-human antibody clones. Furthermore, productive infection of peripheral T lymphocytes with simian immunodeficiency virus (SIV) in vitro was confirmed by intracellular SIV p27 staining. This study could facilitate using Japanese macaques as models for human hematological and immunological disorders and infectious diseases.     

Mitochondrial Remodeling in Endothelial Cells under Cyclic Stretch is Independent of Drp1 Activation

Mitochondria in endothelial cells remodel morphologically when supraphysiological cyclic stretch is exerted on the cells. During remodeling, mitochondria become shorter, but how they do so remains elusive. Drp1 is a regulator of mitochondrial morphologies. It shortens mitochondria by shifting the balance from mitochondrial fusion to fission. In this study, we hypothesized that Drp1 activation is involved in mitochondrial remodeling under supraphysiological cyclic stretch. To verify the involvement of Drp1, its activation was first quantified with Western blotting, but Drp1 was not significantly activated in endothelial cells under supraphysiological cyclic stretch. Next, Drp1 activation was inhibited with Mdivi-1, but this did not inhibit mitochondrial remodeling. Intracellular Ca²⁺ increase activates Drp1 through calcineurin. First, we inhibited the intracellular Ca²⁺ increase with Gd³⁺ and thapsigargin, but this did not inhibit mitochondrial remodeling. Next, we inhibited calcineurin with cyclosporin A, but this also did not inhibit mitochondrial remodeling. These results indicate that mitochondrial remodeling under supraphysiological cyclic stretch is independent of Drp1 activation. In endothelial cells under supraphysiological cyclic stretch, reactive oxygen species (ROS) are generated. Mitochondrial morphologies are remodeled by ROS generation. When ROS was eliminated with N-acetyl-L-cysteine, mitochondrial remodeling was inhibited. Furthermore, when the polymerization of the actin cytoskeleton was inhibited with cytochalasin D, mitochondrial remodeling was also inhibited. These results suggest that ROS and actin cytoskeleton are rather involved in mitochondrial remodeling. In conclusion, the present results suggest that mitochondrial remodeling in endothelial cells under supraphysiological cyclic stretch is induced by ROS in association with actin cytoskeleton rather than through Drp1 activation.     

Crim1C140S mutant mice reveal the importance of cysteine 140 in the internal region 1 of CRIM1 for its physiological functions

Cysteine-rich transmembrane bone morphogenetic protein regulator 1 (CRIM1) is a type I transmembrane protein involved in the organogenesis of many tissues via its interactions with growth factors including BMP, TGF-β, and VEGF. In this study, we used whole-exome sequencing and linkage analysis to identify a novel Crim1 mutant allele generated by ENU mutagenesis in mice. This allele is a missense mutation that causes a cysteine-to-serine substitution at position 140, and is referred to as Crim1^C140S. In addition to the previously reported phenotypes in Crim1 mutants, Crim1^C140S homozygous mice exhibited several novel phenotypes, including dwarfism, enlarged seminal vesicles, and rectal prolapse. In vitro analyses showed that Crim1^C140S mutation affected the formation of CRIM1 complexes and decreased the amount of the overexpressed CRIM1 proteins in the cell culture supernatants. Cys140 is located in the internal region 1 (IR1) of the N-terminal extracellular region of CRIM1 and resides outside any identified functional domains. Inference of the domain architecture suggested that the Crim1^C140S mutation disturbs an intramolecular disulfide bond in IR1, leading to the protein instability and the functional defects of CRIM1. Crim1^C140S highlights the functional importance of the IR1, and Crim1^C140S mice should serve as a valuable model for investigating the functions of CRIM1 that are unidentified as yet.

     

An Insecticidal Alkaloid,Cocculolidine from Cocculus trilobus DC.

Cocculolidine isolated from leaves of Cocculus trilobus DC. (Kamiebi in Japanese), the host plant of Japanese fruit-piercing moths, exhibited insecticidal activity to leaf hoppers (Nephotettix bipunctatus cincticepts Uhler) and Azuki-been weevils (Callosobruchus chinensis Linne), however it had no activity to larvae of the fruit-piercing moth, Oraesia excavata Butler (Akaeguriba in Japanese), suggesting an interesting phenomenon of the host-parasite interrelationship.     

Growth during the early life history of the Pacific tarpon,Megalops cyprinoides

The early growth of the Pacific tarpon,Megalops cyprinoides, was studied by larval otolith analysis and rearing of larvae and juveniles in the laboratory. Morphology of the sagitta, validation of sagittal daily increments, age at the start of metamorphosis, decrement of standard length in early metamorphosis, and growth under rearing conditions are described. The sagitta of fully-grown Pacific tarpon leptocephali were transparent and circular, with regular intervals between the neighboring rings becoming wider at the onset of metamorphosis. Alizarin complexone treatment of larvae confirmed the daily formation of the sagittal rings. Metamorphosis was estimated to start about one month after hatching. After drastic shrinkage during the first several days of metamorphosis, the body length more or less stabilized for one month and then resumed rapid growth. The early growth of Pacific tarpon was divided into four phases as follows: A) leptocephalus positive growth phase; B) leptocephalus negative growth phase; C) sluggish growth phase; and D) juvenile growth phase.     

Relationship between growth temperature of Anacystis nidulans and phase transition temperature of its thylakoid membranes

The temperatures of the lipid phase transition at which the solid phase disappears were determined by using the X-ray diffraction method in thylakoid membranes of the blue-green alga, Anacystis nidulans. The temperatures were determined as 26 and 16°C for cells grown at 38 and 28°C, respectively.     

S-Locus-Specific Glycoproteins Associated with Self-Incompatibility in Brassica campestris

Specific S-glycoproteins were isolated from three Brassica campestriscultivars homozygous with respect to the S-alleles S₈, S₉ andS₁₂. Amino acid sequences of various peptide fragments of theS-proteins were determined using a gas-phase protein sequencer,and a nearly complete amino acid sequence of the S₈-glycoproteinwas determined on the basis of the revised cDNA sequence ofthe B. oleracea S-specific glycoprotein. The lysyl endopeptidasefragments of S₉ and S₁₂-glycoproteins were aligned in comparisonwith the sequence of the S₈-glycoprotein. Although extensivesequence homology was evident among the three S-glycoproteins,the sequences of the middle part were relatively different fromeach other. The numbers and positions of N-glycosylation alsodiffered among the S-glycoproteins of Brassica species. (Received April 20, 1987; Accepted July 29, 1987)