Association between dynapenia and multimorbidity: A systematic review.

Dynapenia and multimorbidity are common health problems affecting older adults. However, few studies have systematically reviewed the association between dynapenia and multimorbidity. Therefore, this systematic review aimed to provide a comprehensive overview of studies on the association between these conditions. We searched four electronic databases for relevant articles published in July 2023. The main inclusion criteria were the following: (1) a description of dynapenia, which indicates loss of muscle strength and (2) a description of multimorbidity with two or more chronic diseases. Five studies met these inclusion criteria. In all five of these studies, the participants were community-dwelling older adults. All the studies showed an association between dynapenia and multimorbidity. The prevalence of dynapenia and multimorbidity ranged from 16% to 25.9%. The results of our systematic review demonstrated that dynapenia in older adults increases the risk of multimorbidity. We propose that interventions and reversible changes in dynapenia can prevent multimorbidity.

Vesicular stomatitis virus pseudotyped with severe acute respiratory syndrome coronavirus spike protein.

Severe acute respiratory syndrome coronavirus (SARS-CoV) contains a single spike (S) protein, which binds to its receptor, angiotensin-converting enzyme 2 (ACE2), induces membrane fusion and serves as a neutralizing antigen. A SARS-CoV-S protein-bearing vesicular stomatitis virus (VSV) pseudotype using the VSVDeltaG* system was generated. Partial deletion of the SARS-CoV-S protein cytoplasmic domain allowed efficient incorporation into VSV particles and led to the generation of a pseudotype (VSV-SARS-St19) at high titre. Green fluorescent protein expression was demonstrated as early as 7 h after infection of Vero E6 cells with VSV-SARS-St19. VSV-SARS-St19 was neutralized by anti-SARS-CoV antibody and soluble ACE2, and its infection was blocked by treatment of Vero E6 cells with anti-ACE2 antibody. These results indicated that VSV-SARS-St19 infection is mediated by SARS-CoV-S protein in an ACE2-dependent manner. VSV-SARS-St19 will be useful for analysing the function of SARS-CoV-S protein and for developing rapid methods of detecting neutralizing antibodies specific for SARS-CoV infection.

Stringent requirement for the C protein of wild-type measles virus for growth both in vitro and in macaques.

The P gene of measles virus (MV) encodes the P protein and three accessory proteins (C, V, and R). However, the role of these accessory proteins in the natural course of MV infection remains unclear. For this study, we generated a recombinant wild-type MV lacking the C protein, called wtMV(C-), by using a reverse genetics system (M. Takeda, K. Takeuchi, N. Miyajima, F. Kobune, Y. Ami, N. Nagata, Y. Suzaki, Y. Nagai, and M. Tashiro, J. Virol. 74:6643-6647). When 293 cells expressing the MV receptor SLAM (293/hSLAM) were infected with wtMV(C-) or parental wild-type MV (wtMV), the growth of wtMV(C-) was restricted, particularly during late stages. Enhanced green fluorescent protein-expressing wtMV(C-) consistently induced late-stage cell rounding and cell death in the presence of a fusion-inhibiting peptide, suggesting that the C protein can prevent cell death and is required for long-term MV infection. Neutralizing antibodies against alpha/beta interferon did not restore the growth restriction of wtMV(C-) in 293/hSLAM cells. When cynomolgus monkeys were infected with wtMV(C-) or wtMV, the number of MV-infected cells in the thymus was >1,000-fold smaller for wtMV(C-) than for wtMV. Immunohistochemical analyses showed strong expression of an MV antigen in the spleen, lymph nodes, tonsils, and larynx of a cynomolgus monkey infected with wtMV but dramatically reduced expression in the same tissues in a cynomolgus monkey infected with wtMV(C-). These data indicate that the MV C protein is necessary for efficient MV replication both in vitro and in cynomolgus monkeys.

Enhancing of measles virus infection by magnetofection.

Magnetofection is a viral and non-viral gene delivery method using polyethyleneimine-conjugated super-paramagnetic nanoparticle under a magnetic field. Previous studies have indicated that magnetofection enhanced the infection of adenoviruses and retroviruses. It is shown that magnetofection enhances the infection of measles virus, a paramyxovirus. When cells expressing a measles virus receptor human SLAM were infected with a measles virus that encodes green fluorescent protein gene, magnetofection enhanced measles virus infection by 30- to 70-fold. The infection of SLAM-negative cells with measles virus was also enhanced by magnetofection, but to a lesser extent. These results indicate that magnetofection could be useful for isolation of measles virus from clinical specimens.

Efficient rescue of measles virus from cloned cDNA using SLAM-expressing Chinese hamster ovary cells.

We here report a highly efficient reverse genetics system for measles virus (MeV), using Chinese hamster ovary cells constitutively expressing a MeV receptor human signaling lymphocyte activation molecule (CHO/hSLAM cells). The recombinant vaccinia virus vTF7-3 that encodes the T7 RNA polymerase under the control of the early/late promoter was used in the system. Replication of vTF7-3 was highly restricted in CHO/hSLAM cells, but the virus could still drive the T7 promoter, allowing us to recover MeV from the transfected cDNA efficiently. With this system the number of infectious centers, in which MeV replication cycles are initiated from transfected cDNAs, was approximately 100 times higher than that with the previous system (. J. Virol. 74, 6643-6647), and the recovery rate was 100%. The wild-type MeV that encodes the lac-Z gene of approximately 3.2kb in length, was easily generated with this CHO/hSLAM system, while such virus could not be recovered with the previous system. Since SLAM acts as a cellular receptor for both MeV vaccine and wild-type strains, the Edmonston vaccine strain was also recovered with this system more efficiently than with any other systems reported previously. Thus, the CHO/hSLAM-based system would expand applications of the MeV reverse genetics by allowing productions of mutant MeVs that have been difficult to generate with less efficient systems.

Cell tropism of wild-type measles virus is affected by amino acid substitutions in the P, V and M proteins, or by a truncation in the C protein.

Two nucleotide differences in the P/C/V and M genes between B95a cell- and Vero cell-isolated wild-type measles viruses (MV) have previously been found from the same patient. The nucleotide difference in the P/C/V gene resulted in an amino acid difference (M175I) in the P and V proteins and a 19 aa deletion in the C protein. The nucleotide difference in the M gene resulted in an amino acid difference (P64H) in the M protein. To verify this result and to examine further whether the amino acid difference or truncation is important for MV cell tropism, recombinant MV strains containing one of the two nucleotide substitutions, or both, were generated. It was found that the P64H substitution in the M protein was important for efficient virus growth and dissemination in Vero cells and that the M175I substitution in the P and V protein or truncation of the C protein was required for optimal growth.

Wild-type measles virus induces large syncytium formation in primary human small airway epithelial cells by a SLAM(CD150)-independent mechanism.

In the natural course of measles virus (MV) infection, epithelial cells are primary targets of MV. However, it has been shown that wild-type MV utilizes signaling lymphocyte activation molecule (SLAM or CD150) as a cellular receptor, which is expressed only in some T and B cells, thymocytes, and dendritic cells. To understand how wild-type MV infects non-lymphoid cells, several non-lymphoid cells were examined for their susceptibility to wild-type MV. Here, we show that wild-type MV can infect primary human small airway epithelial cells (SAEC) and induce formation of large syncytia in vitro. mRNA specific for SLAM was not detected in SAEC, indicating that wild-type MV infects SAEC and induces syncytia formation via a SLAM-independent mechanism.

Measles virus V protein blocks interferon (IFN)-alpha/beta but not IFN-gamma signaling by inhibiting STAT1 and STAT2 phosphorylation.

Measles virus (MV), a member of the family Paramyxoviridae, encodes C and V non-structural proteins. To clarify the functions of MV C and V proteins, HeLa cell lines constitutively expressing C or V protein were established. We found that expression of V protein inhibited interferon (IFN)-alpha/beta signaling but not IFN-gamma signaling. C protein had no inhibitory effect on IFN signaling in our experimental condition. Degradation of selective signal transducers and activators of transcription (STAT) proteins was not observed in HeLa cells expressing V protein. In contrast, tyrosine phosphorylation of both STAT1 and STAT2 was inhibited in these cells after IFN-beta stimulation.

Toward understanding the pathogenicity of wild-type measles virus by reverse genetics.

The Edmonston (Ed) strain of measles virus (MV) isolated in primary human kidney cells in 1954 has long been thought of as a representative MV strain. But this view has been challenged by wild-type MV strains isolated in marmoset B-lymphoblastoid B95a cells. Although the Ed strain is not pathogenic in monkey models, wild-type MV isolated in B95a cells from measles patients induces clinical signs typical of human measles, indicating that wild-type MV retains its pathogenicity. In addition, wild-type MV has restricted cell tropism and replicates only in B95a and some lymphocyte cell lines. This is in sharp contrast to the ability of the Ed strain to replicate in a variety of human cell lines. To understand the molecular basis for the pathogenicity and the cell tropism of wild-type MV, we have established a reverse genetics system based on a highly pathogenic wild-type MV strain (IC-B) isolated in B95a cells. By using this system, we have constructed recombinant wild-type and Ed strains of MV bearing heterologous envelope hemagglutinin (H) proteins, and we have examined roles of the H protein in determining the cell tropism. Our results clearly indicate that the MV cell tropism is determined by not only the H protein, but also other viral proteins. We thus propose the presence of another unidentified MV receptor on the surface of Vero cells.

Recombinant wild-type and edmonston strain measles viruses bearing heterologous H proteins: role of H protein in cell fusion and host cell specificity.

Wild-type measles virus (MV) isolated from B95a cells has a restricted host cell specificity and hardly replicates in Vero cells, whereas the laboratory strain Edmonston (Ed) replicates in a variety of cell types including Vero cells. To investigate the role of H protein in the differential MV host cell specificity and cell fusion activity, H proteins of wild-type MV (IC-B) and Ed were coexpressed with the F protein in Vero cells. Cell-cell fusion occurred in Vero cells when Ed H protein, but not IC-B H protein, was expressed. To analyze the role of H protein in the context of viral infection, a recombinant IC-B virus bearing Ed H protein (IC/Ed-H) and a recombinant Ed virus bearing IC-B H protein (Ed/IC-H) were generated from cloned cDNAs. IC/Ed-H replicated efficiently in Vero cells and induced small syncytia in Vero cells, indicating that Ed H protein conferred replication ability in Vero cells on IC/Ed-H. On the other hand, Ed/IC-H also replicated well in Vero cells and induced small syncytia, although parental Ed induced large syncytia in Vero cells. These results indicated that an MV protein(s) other than H protein was likely involved in determining cell fusion and host cell specificity of MV in the case of our recombinants. SLAM (CDw150), a recently identified cellular receptor for wild-type MV, was not expressed in Vero cells, and a monoclonal antibody against CD46, a cellular receptor for Ed, did not block replication or syncytium formation of Ed/IC-H in Vero cells. It is therefore suggested that Ed/IC-H entered Vero cells through another cellular receptor.

Dephosphorylation failure of tyrosine-phosphorylated STAT1 in IFN-stimulated Sendai virus C protein-expressing cells.

Sendai virus C protein (SeV C) has been reported to counteract the antiviral activities of interferons (IFNs) by inhibiting the expression of IFN-stimulated gene products. In SeV C-expressing cells, formation of an active ISGF3 complex and translocation of STAT1 into the nucleus were not observed. STAT1 was continuously phosphorylated at tyrosine 701 by IFN signaling; however, its serine phosphorylation was suppressed. In addition, tyrosine-phosphorylated STAT1 grew to form abnormally huge complexes. These findings suggest that the counteraction of IFN in SeV C-expressing cells is caused by disordered phosphorylation and dephosphorylation of STAT1.