EVOLUTION OF EQUINE ARTERITIS VIRUS DURING PERSISTENT INFECTION IN THE REPRODUCTIVE TRACT OF THE STALLION AND THE MALE DONKEY

Equine arteritis virus (EAV) establishes persistent infection in the stallion reproductive tract, and the carrier stallion continues to shed virus in semen for weeks to years or lifelong. The objective of this study was to elucidate the intra-host evolution of EAV during persistent infection in stallions. Seven EAV seronegative stallions were experimentally infected with EAV KY84 strain and followed for 726 days post-infection, and sequential clinical samples including semen were collected for virus isolation and next-generation sequencing (NGS). In addition, archived sequential semen samples from two stallions that were naturally infected with EAV KY84 for a long-period (up to 10 years) were also sequenced by NGS. The data demonstrated genetic bottleneck event and selection during acute infection followed by intra-host quasispecies diversification during persistent infection in the stallion reproductive tract. Also, the full-length genome of a novel EAV donkey strain from Chile and a noncytopathic bovine viral diarrhea virus-1 (ncpBVDV-1) strain contaminating rabbit kidney-13 cells were also sequenced by NGS. The EAV donkey strain was genetically distinct but antigenically cross-reacted with EAV antisera, and it was phylogenetically closely related to the South African donkey strain of EAV. Genetic and phylogenetic analyses demonstrated that ncpBVDV-1 belongs to BVDV-1b group

Be-Insertion in Interlanguage: A Topic Marker, A Tense/Agreement Morpheme, or Both?

In L2 production, be-forms are often inserted before thematic verbs (e.g., She is study hard). There have been two hypotheses concerning the function of be-forms, that they are topic markers transferred from L1 or tense/agreement morphemes. This study examines the explanatory adequacy of the two hypotheses by comparing acceptability judgments and self-paced listening between Korean and Russian EFL learners. The results support neither hypothesis completely. Contrary to the Topic Marker Hypothesis, both L2 groups accepted be-insertion after non-topic as well as topic subjects. Against the T/AGR Morpheme Hypothesis, participants also accepted be-insertion after topicalized non-subjects. These results suggest that be-forms can have two functions simultaneously, and thus the two hypotheses must not be considered mutually exclusive. The data also call into question the role of L1 transfer, suggesting that the phenomenon may reflect a universal L2 strategy

Phytopythium leanoi sp. nov. and Phytopythium dogmae sp. nov., Phytopythium species associated with mangrove leaf litter from the Philippines

The genus Phytopythium is a monophyletic taxon of the Peronosporaceae with characteristics intermediate between Phytophthora and Pythium. In the Philippines, reports of Phytopythium are scarce, with the mangrove-swamp-inhabiting species Phytopythium kandeliae being the only species recorded to date. It was the aim of the current study to investigate the diversity of Phytopythium in mangrove habitats in more detail. Based on culture characteristics, morphology, and molecular phylogenetic position, two new species of Phytopythium are described from Philippine mangroves, P. leanoi USTCMS 4102 and P. dogmae USTCMS 4101. Phytopythium leanoi is a species morphologically similar to P. kandeliae, but with the ability to develop gametangia in a homothallic fashion. The other new species, P. dogmae, is characterized by having a short discharge tube, semipapillate to papillate sporangia and frequently exhibiting a clustering of two sporangia per sporangiogenic hypha. With the addition of the two species described in this study, the genus Phytopythium has grown from around 10 to beyond 20 recognized species over the past decade, and it seems likely that several more species of this genus await discovery

Complete Genome Sequence of Noncytopathic Bovine Viral Diarrhea Virus 1 Contaminating a High-Passage RK-13 Cell Line

A high-passage rabbit kidney RK-13 cell line (HP-RK-13[KY], originally derived from the ATCC CCL-37 cell line) used in certain laboratories worldwide is contaminated with noncytopathic bovine viral diarrhea virus (ncpBVDV). On complete genome sequence analysis, the virus strain was found to belong to BVDV group 1b

Promising results from the use of a korean drama to address knowledge, attitudes, and behaviors on school bullying and mental health among Asian American college-aged students

The limited research on bullying, mental health (MH), and help-seeking for Asian American (ASA) college students is concerning due to the public health importance. Korean drama (K-Drama) television shows may be an innovative approach to improve knowledge, attitudes, and behaviors (KAB) on bullying. This study examined whether the KAB about school bullying improved after watching a K-Drama and asked participants about their perspectives of using a K-Drama as an intervention. A convenience sample of college students (n = 118) watched a K-Drama portraying school bullying and MH issues. Pre-/post-tests on KAB on bullying were conducted. Interviews (n = 16) were used to understand their experiences with K-Dramas. The mean age was 22.1 years (1.6 SD), 83.9% were female, and 77.1% were ASAs. Many reported experiences with anxiety (67.8%), depression (38.1%), and school bullying victim experience (40.8%). Post-test scores revealed significant differences in knowledge by most school bullying variables (e.g., victim; witness) and MH issues. There were varying significant findings in post-test scores in attitudes and behaviors by these variables. Participants reported that they “love” the drama, felt an emotional connection, and thought that K-Dramas can be an educational tool for ASAs. K-Dramas may be an effective population-level tool to improve health outcomes among ASAs

Racial Bias Beliefs Related to COVID-19 Among Asian Americans, Native Hawaiians, and Pacific Islanders: Findings From the COVID-19 Effects on the Mental and Physical Health of Asian Americans and Pacific Islanders Survey Study (COMPASS)

Background: During the COVID-19 pandemic, there have been increased reports of racial biases against Asian American and Native Hawaiian and Pacific Islander individuals. However, the extent to which different Asian American and Native Hawaiian and Pacific Islander groups perceive and experience (firsthand or as a witness to such experiences) how COVID-19 has negatively affected people of their race has not received much attention. Objective: This study used data from the COVID-19 Effects on the Mental and Physical Health of Asian Americans and Pacific Islanders Survey Study (COMPASS), a nationwide, multilingual survey, to empirically examine COVID-19-related racial bias beliefs among Asian American and Native Hawaiian and Pacific Islander individuals and the factors associated with these beliefs. Methods: COMPASS participants were Asian American and Native Hawaiian and Pacific Islander adults who were able to speak English, Chinese (Cantonese or Mandarin), Korean, Samoan, or Vietnamese and who resided in the United States during the time of the survey (October 2020 to May 2021). Participants completed the survey on the web, via phone, or in person. The Coronavirus Racial Bias Scale (CRBS) was used to assess COVID-19-related racial bias beliefs toward Asian American and Native Hawaiian and Pacific Islander individuals. Participants were asked to rate the degree to which they agreed with 9 statements on a 5-point Likert scale (ie, 1=strongly disagree to 5=strongly agree). Multivariable linear regression was used to examine the associations between demographic, health, and COVID-19-related characteristics and perceived racial bias. Results: A total of 5068 participants completed the survey (mean age 45.4, SD 16.4 years; range 18-97 years). Overall, 73.97% (3749/5068) agreed or strongly agreed with ≥1 COVID-19-related racial bias belief in the past 6 months (during the COVID-19 pandemic). Across the 9 racial bias beliefs, participants scored an average of 2.59 (SD 0.96, range 1-5). Adjusted analyses revealed that compared with Asian Indians, those who were ethnic Chinese, Filipino, Hmong, Japanese, Korean, Vietnamese, and other or multicultural had significantly higher mean CRBS scores, whereas no significant differences were found among Native Hawaiian and Pacific Islander individuals. Nonheterosexual participants had statistically significant and higher mean CRBS scores than heterosexual participants. Compared with participants aged ≥60 years, those who were younger (aged \u3c30, 30-39, 40-49, and 50-59 years) had significantly higher mean CRBS scores. US-born participants had significantly higher mean CRBS scores than foreign-born participants, whereas those with limited English proficiency (relative to those reporting no limitation) had lower mean CRBS scores. Conclusions: Many COMPASS participants reported racial bias beliefs because of the COVID-19 pandemic. Relevant sociodemographic contexts and pre-existing and COVID-19-specific factors across individual, community, and society levels were associated with the perceived racial bias of being Asian during the pandemic. The findings underscore the importance of addressing the burden of racial bias on Asian American and Native Hawaiian and Pacific Islander communities among other COVID-19-related sequelae

Social Support and Technology Use and Their Association With Mental and Physical Health During the COVID-19 Pandemic Among Asian Americans: The COMPASS Cross-sectional Study

BACKGROUND: The global COVID-19 pandemic disproportionately affected Asian Americans and Pacific Islanders (AAPIs) and revealed significant health disparities with reports of increased discrimination and xenophobia. Among AAPIs, the pandemic exacerbated their social, linguistic, and geographic isolation. Social support may be especially important for AAPIs given the salience of collectivism as a cultural value. Another mechanism for support among AAPIs was technology use, as it is generally widespread among this population. However, older adults may not perceive the same benefits. OBJECTIVE: We examined social support and technology use and their relationships with mental and physical health outcomes through the COVID-19 pandemic among AAPIs. METHODS: Data were drawn from the COVID-19 Effects on the Mental and Physical Health of AAPI Survey Study (COMPASS) for the time period of October 2020 to February 2021. COMPASS was a cross-sectional, multilingual, national survey conducted online, by phone, and in person with AAPI adults who were ≥18 years of age, in collaboration with academic and community partners in the United States. Data were analyzed using multivariable linear regression using the outcome variables of mental and physical health with various predictors such as social support and technology use. We tested for interactions specific to age and ethnicity. RESULTS: Among 4631 AAPIs (mean age 45.9, SD 16.3 years; 2992/4631, 63.1% female), we found that (1) increased social support was associated with better physical health, (2) total social support was positively associated with better mental health, (3) higher technology use was associated with poorer mental health and inversely associated with poorer physical health, (4) the association of technology use with mental health was weaker among those with low social support (vs those with high social support), (5) adults younger than 60 years old (vs ≥60 years old) were more negatively affected with social support and mental health, and (6) Korean Americans appeared to be a high-risk group for poor physical health with increased technology use. CONCLUSIONS: Our paper identified mental and physical health needs along with supportive therapies observed among AAPIs during the pandemic. Future research on how social support can be leveraged, especially among AAPIs younger than 60 years old, and how various types of technology are being utilized are important to guide the recovery efforts to address both mental and physical disparities across communities as a result of the COVID-19 pandemic

Human Plasmablast Migration Toward CXCL12 Requires Glucose Oxidation by Enhanced Pyruvate Dehydrogenase Activity via AKT

Migration of human plasmablast to the bone marrow is essential for the final differentiation of plasma cells and maintenance of effective humoral immunity. This migration is controlled by CXCL12/CXCR4-mediated activation of the protein kinase AKT. Herein, we show that the CXCL12-induced migration of human plasmablasts is dependent on glucose oxidation. Glucose depletion markedly inhibited plasmablast migration by 67%, and the glucose analog 2-deoxyglucose (2-DG) reduced the migration by 53%; conversely, glutamine depletion did not reduce the migration. CXCL12 boosted the oxygen consumption rate (OCR), and 2-DG treatment significantly reduced the levels of all measured tricarboxylic acid (TCA) cycle intermediates. AKT inhibitors blocked the CXCL12-mediated increase of OCR. CXCL12 enhanced the pyruvate dehydrogenase (PDH) activity by 13.5-fold in an AKT-dependent manner to promote mitochondrial oxidative phosphorylation. The knockdown and inhibition of PDH confirmed its indispensable role in CXCL12-induced migration. Cellular ATP levels fell by 91% upon exposure to 2-DG, and the mitochondrial ATP synthase inhibitor oligomycin inhibited CXCL12-induced migration by 85%. Low ATP levels inhibited the CXCL12-induced activation of AKT and phosphorylation of myosin light chains by 42%, which are required for cell migration. Thus, we have identified a mechanism that controls glucose oxidation via AKT signaling and PDH activation, which supports the migration of plasmablasts. This mechanism can provide insights into the proper development of long-lived plasma cells and is, therefore, essential for optimal humoral immunity. To our knowledge, this study is the first to investigate metabolic mechanisms underlying human plasmablast migration toward CXCL12

Equine Arteritis Virus Elicits a Mucosal Antibody Response in the Reproductive Tract of Persistently Infected Stallions

Equine arteritis virus (EAV) has the ability to establish persistent infection in the reproductive tract of the stallion (carrier) and is continuously shed in its semen. We have recently demonstrated that EAV persists within stromal cells and a subset of lymphocytes in the stallion accessory sex glands in the presence of a significant local inflammatory response. In the present study, we demonstrated that EAV elicits a mucosal antibody response in the reproductive tract during persistent infection with homing of plasma cells into accessory sex glands. The EAV-specific immunoglobulin isotypes in seminal plasma included IgA, IgG1, IgG3/5, and IgG4/7. Interestingly, seminal plasma IgG1 and IgG4/7 possessed virus-neutralizing activity, while seminal plasma IgA and IgG3/5 did not. However, virus-neutralizing IgG1 and IgG4/7 in seminal plasma were not effective in preventing viral infectivity. In addition, the serological response was primarily mediated by virus-specific IgM and IgG1, while virus-specific serum IgA, IgG3/5, IgG4/7, and IgG6 isotype responses were not detected. This is the first report characterizing the immunoglobulin isotypes in equine serum and seminal plasma in response to EAV infection. The findings presented herein suggest that while a broader immunoglobulin isotype diversity is elicited in seminal plasma, EAV has the ability to persist in the reproductive tract, in spite of local mucosal antibody and inflammatory responses. This study provides further evidence that EAV employs complex immune evasion mechanisms during persistence in the reproductive tract that warrant further investigation

Equine Arteritis Virus Has Specific Tropism for Stromal Cells and CD8\u3csup\u3e+\u3c/sup\u3e T and CD21\u3csup\u3e+\u3c/sup\u3e B Lymphocytes but Not for Glandular Epithelium at the Primary Site of Persistent Infection in the Stallion Reproductive Tract

Equine arteritis virus (EAV) has a global impact on the equine industry as the causative agent of equine viral arteritis (EVA), a respiratory, systemic, and reproductive disease of equids. A distinctive feature of EAV infection is that it establishes long-term persistent infection in 10 to 70% of infected stallions (carriers). In these stallions, EAV is detectable only in the reproductive tract, and viral persistence occurs despite the presence of high serum neutralizing antibody titers. Carrier stallions constitute the natural reservoir of the virus as they continuously shed EAV in their semen. Although the accessory sex glands have been implicated as the primary sites of EAV persistence, the viral host cell tropism and whether viral replication in carrier stallions occurs in the presence or absence of host inflammatory responses remain unknown. In this study, dual immunohistochemical and immunofluorescence techniques were employed to unequivocally demonstrate that the ampulla is the main EAV tissue reservoir rather than immunologically privileged tissues (i.e., testes). Furthermore, we demonstrate that EAV has specific tropism for stromal cells (fibrocytes and possibly tissue macrophages) and CD8+ T and CD21+ B lymphocytes but not glandular epithelium. Persistent EAV infection is associated with moderate, multifocal lymphoplasmacytic ampullitis comprising clusters of B (CD21+) lymphocytes and significant infiltration of T (CD3+, CD4+, CD8+, and CD25+) lymphocytes, tissue macrophages, and dendritic cells (Iba-1+ and CD83+), with a small number of tissue macrophages expressing CD163 and CD204 scavenger receptors. This study suggests that EAV employs complex immune evasion mechanisms that warrant further investigation