Gamones and mating types in the genus Blepharisma and their possible taxonomic application

SUMMARYMating types I and II ofBlepharisma japonicum v. intermediumexcrete gamones 1 (blepharmone J) and 2 (blepharismone) respectively. The gamone of one type transforms cells of the other type so that they can conjugate with each other. We found that three other species,B. americanum, B. musculusandB. stoltei, have two types of cells homologous to those inB. japonicum; one (type II) excretes a factor which has the same activity as gamone 2 ofB. japonicum, the other (type I) responds to this gamone by cell union. Type I cells of these species also excrete a gamone which induces pairs in type II cells of particular strains. Complementarity for mating is observed in some combinations of the two types.These results indicate that each of the four species has at least one pair of complementary mating types, I and II, with the gamones of the type II's being the same molecule, blepharismone, while gamones of type I's are species- or syngen-specific blepharmone. These generic and specific gamones can be utilized to clarify existing taxonomic and evolutionary questions in the genusBlepharisma

Structural Dynamics of Retroviral Genome and the Packaging

Retroviruses can cause diseases such as AIDS, leukemia, and tumors, but are also used as vectors for human gene therapy. All retroviruses, except foamy viruses, package two copies of unspliced genomic RNA into their progeny viruses. Understanding the molecular mechanisms of retroviral genome packaging will aid the design of new anti-retroviral drugs targeting the packaging process and improve the efficacy of retroviral vectors. Retroviral genomes have to be specifically recognized by the cognate nucleocapsid domain of the Gag polyprotein from among an excess of cellular and spliced viral mRNA. Extensive virological and structural studies have revealed how retroviral genomic RNA is selectively packaged into the viral particles. The genomic area responsible for the packaging is generally located in the 5′ untranslated region (5′ UTR), and contains dimerization site(s). Recent studies have shown that retroviral genome packaging is modulated by structural changes of RNA at the 5′ UTR accompanied by the dimerization. In this review, we focus on three representative retroviruses, Moloney murine leukemia virus, human immunodeficiency virus type 1 and 2, and describe the molecular mechanism of retroviral genome packaging

Capsid Proteins of HIV-1/HIV-2

Timely disassembly of viral core composed of self-assembled capsid (CA) in infected host cells is crucial for retroviral replication. Extensive in vitro studies to date on the self-assembly/disassembly mechanism of human immunodeficiency virus type 1 (HIV-1) CA have revealed its core structure and amino acid residues essential for CA–CA intermolecular interaction. However, little is known about in vitro properties of HIV-2 CA. In this study, we comparatively analyzed the polymerization properties of bacterially expressed HIV-1 and HIV-2 CA proteins. Interestingly, a much higher concentration of NaCl was required for HIV-2 CA to self-assemble than that for HIV-1 CA, but once the polymerization started, the reaction proceeded more rapidly than that observed for HIV-1 CA. Analysis of a chimeric protein revealed that N-terminal domain (NTD) is responsible for this unique property of HIV-2 CA. To further study the molecular basis for different in vitro properties of HIV-1 and HIV-2 CA proteins, we determined thermal stabilities of HIV-1 and HIV-2 CA NTD proteins at several NaCl concentrations by fluorescent-based thermal shift assays. Experimental data obtained showed that HIV-2 CA NTD was structurally more stable than HIV-1 CA NTD. Taken together, our results imply that distinct in vitro polymerization abilities of the two CA proteins are related to their structural instability/stability, which is one of the decisive factors for viral replication potential. In addition, our assay system described here may be potentially useful for searching for anti-CA antivirals against HIV-1 and HIV-2

Phylogenetic Analysis of Vpx/Vpr Expression

Viruses of human immunodeficiency virus type 2 (HIV-2) and some simian immunodeficiency virus (SIV) lineages carry a unique accessory protein called Vpx. Vpx is essential or critical for viral replication in natural target cells such as macrophages and T lymphocytes. We have previously shown that a poly-proline motif (PPM) located at the C-terminal region of Vpx is required for its efficient expression in two strains of HIV-2 and SIVmac, and that the Vpx expression levels of the two clones are significantly different. Notably, the PPM sequence is conserved and confined to Vpx and Vpr proteins derived from certain lineages of HIV-2/SIVs. In this study, Vpx/Vpr proteins from diverse primate lentiviral lineages were experimentally and phylogenetically analyzed to obtain the general expression picture in cells. While both the level and PPM-dependency of Vpx/Vpr expression in transfected cells varied among viral strains, each viral group, based on Vpx/Vpr amino acid sequences, was found to exhibit a characteristic expression profile. Moreover, phylogenetic tree analyses on Gag and Vpx/Vpr proteins gave essentially the same results. Taken together, our study described here suggests that each primate lentiviral lineage may have developed a unique expression pattern of Vpx/Vpr proteins for adaptation to its hostile cellular and species environments in the process of viral evolution

Effects of scattering and dust grain size on the temperature structure of protoplanetary discs: A three-layer approach

The temperature in the optically thick interior of protoplanetary discs is essential for the interpretation of millimeter observations of the discs, for the vertical structure of the discs, for models of the disc evolution and the planet formation, and for the chemistry in the discs. Since large icy grains have a large albedo even in the infrared, the effect of scattering of the diffuse radiation in the discs on the interior temperature should be examined. We have performed a series of numerical radiation transfer simulations including isotropic scattering by grains with various typical sizes for the diffuse radiation as well as for the incident stellar radiation. We also have developed an analytic model including isotropic scattering to understand the physics concealed in the numerical results. With the analytic model, we have shown that the standard two-layer approach is valid only for grey opacity (i.e. grain size \ga10 \micron) even without scattering. A three-layer interpretation is required for grain size \la10 \micron. When the grain size is 0.1--10 \micron, the numerical simulations show that isotropic scattering reduces the temperature of the disc interior. This reduction is nicely explained by the analytic three-layer model as a result of the energy loss by scatterings of the incident stellar radiation and of the warm diffuse radiation in the disc atmosphere. For grain size \ga10 \micron (i.e. grey scattering), the numerical simulations show that isotropic scattering does not affect the interior temperature. This is nicely explained by the analytic two-layer model; the energy loss by scattering in the disc atmosphere is exactly offset by the "green-house effect" due to scattering of the cold diffuse radiation in the interior.Comment: MNRAS accepte

Pharmacokinetics of Chiral Dendrimer-Triamine-Coordinated Gd-MRI Contrast Agents Evaluated by in Vivo MRI and Estimated by in Vitro QCM

Recently, we developed novel chiral dendrimer-triamine-coordinated Gd-MRI contrast agents (Gd-MRI CAs), which showed longitudinal relaxivity (r1) values about four times higher than that of clinically used Gd-DTPA (Magnevist®, Bayer). In our continuing study of pharmacokinetic differences derived from both the chirality and generation of Gd-MRI CAs, we found that the ability of chiral dendrimer Gd-MRI CAs to circulate within the body can be directly evaluated by in vitro MRI (7 T). In this study, the association constants (Ka) of chiral dendrimer Gd-MRI CAs to bovine serum albumin (BSA), measured and calculated with a quartz crystal microbalance (QCM) in vitro, were found to be an extremely easy means for evaluating the body-circulation ability of chiral dendrimer Gd-MRI CAs. The Ka values of S-isomeric dendrimer Gd-MRI CAs were generally greater than those of R-isomeric dendrimer Gd-MRI CAs, which is consistent with the results of our previous MRI study in vivo

Career Decision-Making Self-Efficacy of Freshmen and Sophomores at “A” University during the COVID-19 Pandemic

本稿では，コロナ禍におけるA 大学の大学1・2 年生（大学生低学年）の進路選択セルフ・エフィカシーについて議論する。パネル調査の結果，次の2 つのことが示された。（1）COVID-19 によってもたらされた雇用状況に対する不安は，大学1・2 年生の進路選択セルフ・エフィカシーの向上を阻害している。（2）雇用状況の悪化の下でも，大学1・2 年生の女子学生の進路選択セルフ・エフィカシーは高まっていることが示唆された。We discuss career decision-making self-efficacy (CDMSE) of freshmen and sophomores during the COVID-19 Pandemic. A panel survey with questionnaires (two-way ANOVA with replications) was conducted. The following two results were indicated: (1) the anticipation of a downward trend in employment during the COVID-19 Pandemic is preventing freshmen and sophomores from increasing CDMSE, (2) in spite of the downward trend, first year and second year female students have increased CDMSE

Different interaction between HIV-1 Vif and its cellular target proteins APOBEC3G/APOBEC3F

We examined a series of site-directed point mutants of human immunodeficiency virus type 1 (HIV-1) Vif for their interaction with cellular anti-viral factors APOBEC3G/APOBEC3F. Mutant viruses that display growth-defect in H9 cells did not counteract effectively APOBEC3G and/or APOBEC3F without exception, as monitored by single-cycle infectivity assays. While growth-defective mutants of Vif C-terminal region were unable to suppress APOBEC3G/APOBEC3F, some N-terminal region mutants did neutralize one of APOBEC3G/APOBEC3F. These data have suggested that members of APOBEC3 family other than APOBEC3G/APOBEC3F are not important for anti-HIV-1 activity. Furthermore, APOPEC3G/APOBEC3F were found to differently associate with Vif in virions as analyzed by equilibrium density centrifugation. Taken together, these results indicated that interaction of HIV-1 Vif and APOBEC3G is distinct from that between Vif and APOBEC3F