Virus-Like Vesicles of Kaposi's Sarcoma-Associated Herpesvirus Activate Lytic Replication by Triggering Differentiation Signaling.

Virus-like vesicles (VLVs) are membrane-enclosed vesicles that resemble native enveloped viruses in organization but lack the viral capsid and genome. During the productive infection of tumor-associated gammaherpesviruses, both virions and VLVs are produced and are released into the extracellular space. However, studies of gammaherpesvirus-associated VLVs have been largely restricted by the technical difficulty of separating VLVs from mature virions. Here we report a strategy of selectively isolating VLVs by using a Kaposi's sarcoma-associated herpesvirus (KSHV) mutant that is defective in small capsid protein and is unable to produce mature virions. Using mass spectrometry analysis, we found that VLVs contained viral glycoproteins required for cellular entry, as well as tegument proteins involved in regulating lytic replication, but lacked capsid proteins. Functional analysis showed that VLVs induced the expression of the viral lytic activator RTA, initiating KSHV lytic gene expression. Furthermore, employing RNA sequencing, we performed a genomewide analysis of cellular responses triggered by VLVs and found that PRDM1, a master regulator in cell differentiation, was significantly upregulated. In the context of KSHV replication, we demonstrated that VLV-induced upregulation of PRDM1 was necessary and sufficient to reactivate KSHV by activating its RTA promoter. In sum, our study systematically examined the composition of VLVs and demonstrated their biological roles in manipulating host cell responses and facilitating KSHV lytic replication.IMPORTANCE Cells lytically infected with tumor-associated herpesviruses produce a high proportion of virus-like vesicles (VLVs). The composition and function of VLVs have not been well defined, largely due to the inability to efficiently isolate VLVs that are free of virions. Using a cell system capable of establishing latent KSHV infection and robust reactivation, we successfully isolated VLVs from a KSHV mutant defective in the small capsid protein. We quantitatively analyzed proteins and microRNAs in VLVs and characterized the roles of VLVs in manipulating host cells and facilitating viral infection. More importantly, we demonstrated that by upregulating PRDM1 expression, VLVs triggered differentiation signaling in targeted cells and facilitated viral lytic infection via activation of the RTA promoter. Our study not only demonstrates a new strategy for isolating VLVs but also shows the important roles of KSHV-associated VLVs in intercellular communication and the viral life cycle.

Rapid Detection and Inhibition of SARS-CoV-2-Spike Mutation-Mediated Microthrombosis.

Activation of endothelial cells following severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is thought to be the primary driver for the increasingly recognized thrombotic complications in coronavirus disease 2019 patients, potentially due to the SARS-CoV-2 Spike protein binding to the human angiotensin-converting enzyme 2 (hACE2). Vaccination therapies use the same Spike sequence or protein to boost host immune response as a protective mechanism against SARS-CoV-2 infection. As a result, cases of thrombotic events are reported following vaccination. Although vaccines are generally considered safe, due to genetic heterogeneity, age, or the presence of comorbidities in the population worldwide, the prediction of severe adverse outcome in patients remains a challenge. To elucidate Spike proteins underlying patient-specific-vascular thrombosis, the human microcirculation environment is recapitulated using a novel microfluidic platform coated with human endothelial cells and exposed to patient specific whole blood. Here, the blood coagulation effect is tested after exposure to Spike protein in nanoparticles and Spike variant D614G in viral vectors and the results are corroborated using live SARS-CoV-2. Of note, two potential strategies are also examined to reduce blood clot formation, by using nanoliposome-hACE2 and anti-Interleukin (IL) 6 antibodies.

[Temporal and spacial expression of LIM mineralization protein 1 during rat pulp injury and reparation].

PURPOSE: To investigate the expression and localization of LIM mineralization protein 1(LMP-1) during rat pulp injury and reparation. METHODS: Dental pulp injury models were established in maxillary first molars on one side of 12 Wistar rats, the isonym healthy teeth on the opposite side were used as the controls. Immunohistochemistry technique was used to observe the expression of LMP-1 at 1, 3 and 7 day after pulp injury. The results of staining were analyzed by Image-Pro Plus 6.0 and SPSS 17.0 software package. RESULTS: LMP-1 was not detected in normal rat dental pulp. Positive staining of LMP-1 was found in odontoblasts and some dental pulp cells at 1 day after pulp injury. The expression of LMP-1 was converged at cell proliferation zone under the injury site at 3 and 7 day after injury. CONCLUSIONS: LMP-1 might play a role in the dental pulp cell proliferation and differentiation and reparative dentin formation during pulp injury and reparation.

Comparison of Cervical Spine Anatomy in Calves, Pigs and Humans.

BACKGROUND CONTEXT: Animals are commonly used to model the human spine for in vitro and in vivo experiments. Many studies have investigated similarities and differences between animals and humans in the lumbar and thoracic vertebrae. However, a quantitative anatomic comparison of calf, pig, and human cervical spines has not been reported. PURPOSE: To compare fundamental structural similarities and differences in vertebral bodies from the cervical spines of commonly used experimental animal models and humans. STUDY DESIGN: Anatomical morphometric analysis was performed on cervical vertebra specimens harvested from humans and two common large animals (i.e., calves and pigs). METHODS: Multiple morphometric parameters were directly measured from cervical spine specimens of twelve pigs, twelve calves and twelve human adult cadavers. The following anatomical parameters were measured: vertebral body width (VBW), vertebral body depth (VBD), vertebral body height (VBH), spinal canal width (SCW), spinal canal depth (SCD), pedicle width (PW), pedicle depth (PD), pedicle inclination (PI), dens width (DW), dens depth (DD), total vertebral width (TVW), and total vertebral depth (TVD). RESULTS: The atlantoaxial (C1-2) joint in pigs is similar to that in humans and could serve as a human substitute. The pig cervical spine is highly similar to the human cervical spine, except for two large transverse processes in the anterior regions ofC4-C6. The width and depth of the calf odontoid process were larger than those in humans. VBW and VBD of calf cervical vertebrae were larger than those in humans, but the spinal canal was smaller. Calf C7 was relatively similar to human C7, thus, it may be a good substitute. CONCLUSION: Pig cervical vertebrae were more suitable human substitutions than calf cervical vertebrae, especially with respect to C1, C2, and C7. The biomechanical properties of nerve vascular anatomy and various segment functions in pig and calf cervical vertebrae must be considered when selecting an animal model for research on the spine.

[A comparative study of split-root and medial resistance removal in extraction of medially impacted tooth].

PURPOSE: To compare the operating time, root fracture and postoperative complications between split-root extraction and medial resistance removal in extraction of mandibular small-angle impacted third molars, to evaluate the advantages of split-root extraction in medially impacted tooth extraction. METHODS: Forty male patients with bilaterally mandibular medial small-angle impacted third molars, having multiple roots in panoramic films, were selected. The impacted teeth on one side were extracted by using split-root method, while the similar impacted teeth on the other side were extracted by using medial resistance removal method. The operating time, root fracture, postoperative pain, facial edema, and mouth opening were recorded. SPSS11.5 software package was used and paired t test was performed to analyze the data. RESULTS: There were significant differences in operating time and root fracture between the two methods (P<0.05). In the split-root group, the operating time was shorter and root fracture were less. After 24 hours, facial edema, limited mouth opening, and pain of the split-root group was less severe than that of the medial resistance removal group, differences were significant(P<0.05). After 72 hours, limited mouth opening of the split-root group was milder than that of the resistance removal group, the difference was significant (P<0.05). After 120 hours, there was no significant difference of any complications between the two groups. CONCLUSIONS: In extraction of mandibular medial small-angle impacted third molars with multiple roots, the operating time of split-root extraction is shorter. The root fracture possibility of split-root extraction is smaller, and postoperative complications are less common.