Characterizing the splice map of Turkey Hemorrhagic Enteritis Virus.

BACKGROUND: Hemorrhagic enteritis, caused by Turkey Hemorrhagic Enteritis Virus (THEV), is a disease affecting turkey poults characterized by immunosuppression and bloody diarrhea. An avirulent THEV strain that retains the immunosuppressive ability is used as a live vaccine. Characterizing the splice map of THEV is an essential step that would allow studies of individual genes mediating its immunosuppressive functions. We used RNA sequencing to characterize the splice map of THEV for the first time, providing key insights into the THEV gene expression and mRNA structures. METHODS: After infecting a turkey B-cell line with the vaccine strain, samples in triplicates were collected at 4-, 12-, 24-, and 72-hours post-infection. Total RNA was extracted, and poly-A-tailed mRNA sequenced. Reads were mapped to the THEV genome after trimming and transcripts assembled with StringTie. We performed PCR of THEV cDNA, cloned the PCR products, and used Sanger sequencing to validate all identified splice junctions. RESULTS: Researchers previously annotated the THEV genome as encoding 23 open reading frames (ORFs). We identified 29 spliced transcripts from our RNA sequencing data, all containing novel exons although some exons matched some previously annotated ORFs. The three annotated splice junctions were also corroborated by our data. During validation we identified five additional unique transcripts, a subset of which were further validated by 3' rapid amplification of cDNA ends (3' RACE). Thus, we report that the genome of THEV contains 34 transcripts with the coding capacity for all annotated ORFs. However, we found six of the previously annotated ORFs to be truncated ORFs on the basis of the identification of an in-frame upstream start codon or the detection of additional coding exons. We also identified three of the annotated ORFs with longer or shorter isoforms, and seven novel unannotated ORFs that could potentially be translated; although it is beyond the scope of this manuscript to investigate whether they are translated. CONCLUSIONS: Similar to human adenoviruses, all THEV transcripts are spliced and organized into five transcription units under the control of their cognate promoters. The genes are expressed under temporal regulation and THEV also produces multiple distinctly spliced transcripts that code for the same protein. Studies of the newly identified potential proteins should be urgently performed as these proteins may have roles in THEV-induced immunosuppression. Also, knowing the splicing of THEV genes should be invaluable to future research focusing on studying THEV genes, as this will allow accurate cloning of the mRNAs.

Changes in Attitudes and Barriers to Seasonal Influenza Vaccination from 2007 to 2023.

This study investigated how factors and barriers to flu vaccination among college students has changed over the past 16 years. Data were collected from 440 students using a survey and compared to previous data from the same university. Respondents were also asked about their experiences with Covid-19 and its effect on their intent to vaccinate. We found that vaccination rates had increased from 12.4 to 30.5%. Among the unvaccinated, expense, fear of getting influenza from vaccination, fear of side effects, and lack of information have decreased by 28%, 20%, 17%, and 15% respectively. Time, convenience, and perceived risk are still significant barriers to vaccination. Students are getting more encouragement to vaccinate from their health care providers and parents, but it is becoming less effective. The Covid-19 pandemic has changed vaccine attitudes and vaccine fatigue has been a large contributor. Additionally, political affiliation has become a predictor of flu vaccine uptake with conservatives being less likely to vaccinate. There has also been a shift in motivation from concern for personal safety to concern for public safety.

A Randomized Study to Determine the Effect of a Culturally Focused Video Intervention on Improving HPV Vaccine Intentions in a Christian Population in the United States.

Safe and effective vaccines have been developed that protect against high-risk strains of HPV, but uptake is relatively low. We previously identified factors such as sexual attitudes and HPV knowledge that impact the intent of Christian parents to vaccinate their children against HPV. We hypothesized that culturally specific interventions in the form of short videos would be effective at improving HPV vaccine intentions and attitudes. We made three short educational videos, one with a Christian focus, one informational about HPV, and one control. Videos were distributed electronically with accompanying surveys, and responses were measured before and after watching a randomly selected video. The religious-focused and educational interventions significantly (p < 0.0001, p = 0.0015) improved intentions towards HPV vaccination. The religiously-focused video also significantly diminished the belief that the HPV vaccine is unnecessary because of a family's values (p = 0.014). Parents significantly credited both interventions with improving their intent to vaccinate their children against HPV (p < 0.001 for both). These results suggest that culturally focused educational interventions are effective at influencing vaccine intentions and attitudes, even when those are based on religious or cultural feelings. Highly specific interventions are likely to be necessary for optimal improvement in vaccine hesitancy.

Synthetic Pharmacotherapy for Systemic Lupus Erythematosus: Potential Mechanisms of Action, Efficacy, and Safety.

The pharmacological treatment of systemic lupus erythematosus (SLE) aims to decrease disease activity, progression, systemic compromise, and mortality. Among the pharmacological alternatives, there are chemically synthesized drugs whose efficacy has been evaluated, but which have the potential to generate adverse events that may compromise adherence and response to treatment. Therapy selection and monitoring will depend on patient characteristics and the safety profile of each drug. The aim of this review is to provide a comprehensive understanding of the most important synthetic drugs used in the treatment of SLE, including the current treatment options (mycophenolate mofetil, azathioprine, and cyclophosphamide), review their mechanism of action, efficacy, safety, and, most importantly, provide monitoring parameters that should be considered while the patient is receiving the pharmacotherapy.

EBI2 expression in B lymphocytes is controlled by the Epstein-Barr virus transcription factor, BRRF1 (Na), during viral infection.

Epstein-Barr virus-induced gene 2 (EBI2) is an important chemotactic receptor that is involved in proper B-cell T-cell interactions. Epstein-Barr virus (EBV) has been shown to upregulate this gene upon infection of cell lines, but the timing and mechanism of this upregulation, as well as its importance to EBV infection, remain unknown. This work investigated EBV's manipulation of EBI2 expression of primary naive B cells. EBV infection induces EBI2 expression resulting in elevated levels of EBI2 after 24 h until 7 days post-infection, followed by a dramatic decline (P=0.027). Increased EBI2 expression was not found in non-specifically stimulated B cells or when irradiated virus was used. The EBV lytic gene BRRF1 exhibited a similar expression pattern to EBI2 (R2=0.4622). BRRF1-deficient EBV could not induce EBI2. However, B cells transduced with BRRF1 showed elevated expression of EBI2 (P=0.042), a result that was not seen with transduction of a different EBV lytic transfection factor, BRLF1. Based on these results, we conclude that EBI2 expression is directly influenced by EBV infection and that BRRF1 is necessary and sufficient for EBI2 upregulation during infection.

Piracy on the molecular level: human herpesviruses manipulate cellular chemotaxis.

Cellular chemotaxis is important to tissue homeostasis and proper development. Human herpesvirus species influence cellular chemotaxis by regulating cellular chemokines and chemokine receptors. Herpesviruses also express various viral chemokines and chemokine receptors during infection. These changes to chemokine concentrations and receptor availability assist in the pathogenesis of herpesviruses and contribute to a variety of diseases and malignancies. By interfering with the positioning of host cells during herpesvirus infection, viral spread is assisted, latency can be established and the immune system is prevented from eradicating viral infection.

Effects of religious and cultural beliefs on vaccine attitudes in a Hispanic immigrant population in the United States.

Hispanic Immigrants (HI) have lower vaccination rates than their non-Hispanic white counterparts. Culturally appropriate interventions are more influential in evoking change among viewers; therefore, it is important to understand the cultural factors of specific ethnic groups. In this study, we identify cultural barriers to vaccination of Hispanic Immigrants. An electronic survey was administered among HI parents living in the United States. Using structural equation modeling, we found that high religious practice and positive religious beliefs towards vaccination correspond with positive vaccine attitudes (+0.20 and +0.587, respectively). Trust in institutions is strongly correlated with strong positive vaccine attitudes (+0.734). While trusting one's folk practitioner more than a medical doctor leads to negative vaccine attitudes (-0.596), the use of home remedies is associated with positive vaccine attitudes (+0.486). The cultural competence of a medical practitioner, especially regarding folk medicine, is essential to lower cultural barriers HI patients face in vaccination.

The HIV-1 vpr R77Q Mutant Induces Apoptosis, G2 Cell Cycle Arrest, and Lower Production of Pro-Inflammatory Cytokines in Human CD4+ T Cells.

Acquired immunodeficiency syndrome (AIDS) occurs when HIV depletes CD4+ helper T cells. Some patients develop AIDS slowly or not at all, and are termed long-term non-progressors (LTNP), and while mutations in the HIV-1 Viral Protein R (vpr) gene such as R77Q are associated with LTNP, mechanisms for this correlation are unclear. This study examines the induction of apoptosis, cell cycle arrest, and pro-inflammatory cytokine release in the HUT78 T cell line following infection with replication-competent wild-type strain NL4-3, the R77Q mutant, or a vpr Null mutant. Our results show a significant enhancement of apoptosis and G2 cell cycle arrest in HUT78 cells infected with R77Q, but not with WT NL4-3 or the vpr Null strain. Conversely, HUT78 cells infected with the WT virus show higher levels of necrosis. We also detected lower TNF and IL-6 release after infection with R77Q vs. WT. The apoptotic phenotype was also seen in the CEM cell line and in primary CD4+ T cells. Protein expression of the R77Q vpr variant was low compared to WT vpr, but expression levels alone cannot explain these phenotypes because the Null virus did not show apoptosis or G2 arrest. These results suggest that R77Q triggers a non-inflammatory apoptotic pathway that attenuates inflammation, possibly contributing to LTNP.

Comparison of B-Cell Lupus and Lymphoma Using a Novel Immune Imbalance Transcriptomics Algorithm Reveals Potential Therapeutic Targets.

BACKGROUND/OBJECTIVES: Systemic lupus erythematosus (lupus) and B-cell lymphoma (lymphoma) co-occur at higher-than-expected rates and primarily depend on B cells for their pathology. These observations implicate shared inflammation-related B cell molecular mechanisms as a potential cause of co-occurrence. METHODS: We consequently implemented a novel Immune Imbalance Transcriptomics (IIT) algorithm and applied IIT to lupus, lymphoma, and healthy B cell RNA-sequencing (RNA-seq) data to find shared and contrasting mechanisms that are potential therapeutic targets. RESULTS: We observed 7143 significantly dysregulated genes in both lupus and lymphoma. Of those genes, we found 5137 to have a significant immune imbalance, defined as a significant dysregulation by both diseases, as analyzed by IIT. Gene Ontology (GO) term and pathway enrichment of the IIT genes yielded immune-related "Neutrophil Degranulation" and "Adaptive Immune System", which validates that the IIT algorithm isolates biologically relevant genes in immunity and inflammation. We found that 344 IIT gene products are known targets for established and/or repurposed drugs. Among our results, we found 48 known and 296 novel lupus targets, along with 151 known and 193 novel lymphoma targets. Known disease drug targets in our IIT results further validate that IIT isolates genes with disease-relevant mechanisms. CONCLUSIONS: We anticipate the IIT algorithm, together with the shared and contrasting gene mechanisms uncovered here, will contribute to the development of immune-related therapeutic options for lupus and lymphoma patients.

Comparison of Predictive Factors of Flu Vaccine Uptake Pre- and Post-COVID-19 Using the NIS-Teen Survey.

BACKGROUND: Seasonal influenza vaccination rates are very low among teenagers. OBJECTIVES: We used publicly available data from the NIS-Teen annual national immunization survey to explore factors that influence the likelihood of a teen receiving their seasonal flu shot. METHODS: Traditional stepwise multivariable regression was used in tandem with machine learning to determine the predictive factors in teen vaccine uptake. RESULTS AND CONCLUSIONS: Age was the largest predictor, with older teens being much less likely to be vaccinated than younger teens (97.48% compared to 41.71%, p < 0.0001). Provider participation in government programs such as Vaccines for Children and the state vaccine registry positively impacts vaccine uptake (p < 0.0001). Identifying as non-Hispanic Black was a small, negative predictor of teen vaccine uptake (78.18% unvaccinated compared to 73.78% of White teens, p < 0.0001). The state quartile for COVID-19 vaccine uptake also strongly predicted flu vaccine uptake, with the upper quartile of state COVID-19 vaccine uptake being significantly more likely to also get vaccinated for influenza (76.96%, 74.94%, 74.55%, and 72.97%, p < 0.0001). Other significant factors are the number of providers, education of the mother, poverty status, and having a mixed provider facility type. Additionally, the multivariable regression analysis revealed little difference in the predictive factors of vaccine uptake between pre- and post-pandemic datasets.

Effects of religion, politics and distance to providers on HPV vaccine attitudes and intentions of parents in rural Utah.

PURPOSE: Human papillomavirus (HPV) causes an estimated 300,000 high grade cervical dysplasias and 36,000 preventable cancers each year in the United States alone. Despite having a safe, effective and long lasting vaccine since 2006, the rate of uptake has been suboptimal, particularly in rural areas. In 2020, Utah ranked near last in teenage HPV vaccination rates with rural areas trailing urban areas by about 18 percent. In this study, we identified factors that affect the intent of rural Utah parents to vaccinate their children against HPV. METHODS: A survey was distributed electronically to Utah residents in rural areas. Recruitment was carried out through targeted advertising, community organizations, and professional survey panels. The survey was open from Nov. 15, 2022 to April 15, 2023. A total of 410 respondents were used for analysis. Survey results were analyzed using exploratory factor analysis, confirmatory factor analysis, and structural equation modeling. FINDINGS: Distance to care was shown to negatively influence direct intent to vaccinate, while trust in government, general vaccine attitudes, and HPV knowledge positively influence intent to vaccinate. It was found that religious practice decreased vaccine hesitancy while cautious sexual attitudes, distance to care, and general negative vaccine attitudes increased vaccine hesitancy. Conservative political identity and high income were both shown to decrease vaccine hesitancy as covariates.

Cytokine inhibition as a strategy for treating systemic lupus erythematosus.

Cytokines regulate and control the immune system. In systemic lupus erythematosus, several of these cytokines are overexpressed and contribute to the pathogenesis of the disease. Cytokine inhibition has been successfully used to treat other rheumatic and autoimmune diseases, and several cytokines are currently being investigated to determine whether inhibition would be therapeutic in lupus. The cytokines discussed in this review have all undergone clinical trials, and include TNF-α, IL-1, IL-6, IL-10, IL-15, IL-17, IL-18 and IL-23. Inhibition of the majority of these targets was safe and showed some efficacy in treating lupus. Cytokine inhibition strategies have just started to realize their potential for the treatment of this difficult disease, and show great promise for the future.

Association between Religious Beliefs and HPV Vaccination Attitudes among College Students.

OBJECTIVE AND PARTICIPANTS: The authors sought an updated examination of attitudes toward Human Papillomavirus (HPV) catch-up vaccination among college students at a private religious university. METHODS: A total of 1557 college students completed a 62-question survey of religious and HPV vaccination attitudes during the fall of 2021. Students' willingness to receive catch-up HPV vaccination and willingness to vaccinate a future child against HPV were recorded. RESULTS: Of the 46.8% of students who reported being unvaccinated or unaware of vaccination status, ~26% reported being uninterested in receiving catch-up HPV vaccination; ~22% of all students surveyed reported being unwilling to vaccinate a future child against HPV. The strongest predictors of vaccine hesitancy included religious concerns about sexual abstinence and safety concerns. CONCLUSIONS: College health professionals can increase the rate of HPV vaccination among college students and subsequent future generations by addressing the safety and utility of the vaccine regardless of intentions for sexual abstinence prior to marriage. Additionally, rather than a uniform approach to all students who self-identify as Christian, an effort to identify and discuss the unique religiously influenced beliefs of individual students is recommended when discussing HPV vaccination.

A Novel Application of Spinning Disk Technology to Collect Plasma from Whole Blood Prior to Quantifying Plasma RNA.

The spinning disk technology has previously been utilized to isolate bacterial components from blood in hours instead of days. We hypothesized that this platform could be applied as an alternative approach to isolating plasma RNA from a whole blood sample. We consequently tested the efficacy of the spinning disk technology to extract plasma from whole blood upstream of RNA isolation and analysis. To do so, we collected plasma using either the spinning disk or the typical two-spin centrifuge method. We found that the spinning disk method results in significantly more hemolysis during collection than the conventional two-spin centrifuge method. However, when plasma RNA recovered from both collection methods was quantified using quantitative Real-Time Polymerase Chain Reaction (qRT-PCR), we found that the spinning disk method yielded a higher plasma RNA concentration than the two-spin centrifuge method. This suggests that the spinning disk may be an efficient alternative method to recover plasma RNA. Further work is needed to determine whether red blood cell RNA contamination is present in the plasma RNA extracted from spinning disk-processed plasma.

Factors Affecting Vaccine Attitudes Influenced by the COVID-19 Pandemic.

The development of vaccines has significantly contributed to the success of disease prevention. However, there has been a sharp decline in immunization rates since COVID-19 spread globally. Seemingly overnight, the world shut down and most non-essential medical procedures were postponed. Since the COVID-19 vaccine became available, and the world started going back to normal these vaccine rates have not recovered. In this paper, we review the published literature to explore how convenience factors, perceived risk of vaccination, media or anti-vaccination ideals/movements, and healthcare professionals affect an individual's compliance to be vaccinated to better understand the factors that contribute to the change in overall vaccination rates.

Decrease in Overall Vaccine Hesitancy in College Students during the COVID-19 Pandemic.

The COVID-19 pandemic changed our world as we know it and continues to be a global problem three years since the pandemic began. Several vaccines were produced, but there was a considerable amount of societal turmoil surrounding them that has affected the way people view not only COVID-19 vaccines but all vaccines. We used a survey to compare how attitudes towards vaccination have changed in college students during the pandemic. An initial survey was administered in 2021, then a follow-up in 2022. Out of 316 respondents who answered the first survey, 192 completed the follow-up. The survey was designed to measure trends in changes to vaccine attitudes since the COVID-19 pandemic began. By comparing the first survey in 2021 and the follow-up, we found that roughly 55% of respondents' vaccine attitudes did not change, roughly 44% of respondents' attitudes towards vaccines became more positive, and only about 1% of the respondents' vaccine attitudes became more negative. Improved view of vaccines was associated with political views and increased trust in medicine and the healthcare system. Worsened opinions of vaccines were associated with a belief that the COVID-19 vaccine affected fertility.

Effects of IRF5 lupus risk haplotype on pathways predicted to influence B cell functions.

Both genetic and environmental interactions affect systemic lupus erythematosus (SLE) development and pathogenesis. One known genetic factor associated with lupus is a haplotype of the interferon regulatory factor 5 (IRF5) gene. Analysis of global gene expression microarray data using gene set enrichment analysis identified multiple interferon- and inflammation-related gene sets significantly overrepresented in cells with the risk haplotype. Pathway analysis using expressed genes from the significant gene sets impacted by the IRF5 risk haplotype confirmed significant correlation with the interferon pathway, Toll-like receptor pathway, and the B-cell receptor pathway. SLE patients with the IRF5 risk haplotype have a heightened interferon signature, even in an unstimulated state (P = 0.011), while patients with the IRF5 protective haplotype have a B cell interferon signature similar to that of controls. These results identify multiple genes in functionally significant pathways which are affected by IRF5 genotype. They also establish the IRF5 risk haplotype as a key determinant of not only the interferon response, but also other B-cell pathways involved in SLE.

Influences on Attitudes Regarding COVID-19 Vaccination in Germany.

Trust in institutions and democracy may be a major contributor to the willingness to be vaccinated. We investigated these factors and others with regard to COVID-19 vaccine uptake in Germany. Even though effective vaccination is a major contributor to slowing down the current pandemic, vaccine hesitancy remains a major challenge. To analyze attitudes toward vaccine hesitancy, a web-based cross-sectional survey was used to understand and describe the influences of attitudes about vaccination against COVID-19 in the German population. A descriptive analysis for the entire dataset was carried out, and multiple proportional odds regression, path model, and structural equation modeling (SEM) were subsequently used to analyze any relationship between latent variables and COVID-19 vaccine acceptance. In total, 1092 responses from across Germany were analyzed. SEM modeling revealed that trust in institutions, trust in non-pharmaceutical interventions, and various demographic factors were associated with intent to vaccinate. Descriptive analysis and multiple proportional odds regression confirmed that a history of influenza vaccination and level of satisfaction with democratic institutions were highly predictive (p < 0.05) for COVID-19 vaccine acceptance. Additionally, social determinants of health such as gender, age, number of children in the family, and the degree of satisfaction with life were also predictors (p < 0.05) for COVID-19 vaccine acceptance. Results also demonstrated a significant relationship between receiving the flu vaccine and acceptance of the COVID-19 vaccination. Governments that provide COVID-19 vaccines and control messaging should strive for trust and transparency to maximize vaccine uptake. Government-based vaccine measures should also involve measures to communicate trust in democratic and scientific institutions.

Cautious Sexual Attitudes Diminish Intent to Vaccinate Children against HPV in Utah.

Although most Human papillomavirus (HPV) infections are mild and are cleared by the immune system, some high-risk HPV strains can cause various cancers. Vaccines have been developed that protect against high-risk HPV strains. HPV vaccines have been approved for use by the CDC (Centers for Disease Control and Prevention) and are recommended for everyone aged 11-26. Despite the availability of safe and effective vaccines, uptake is low. HPV vaccine uptake has been extensively studied on a national and international level, but less is known about vaccine acceptance on a state or local level. The state of Utah, in the United States of America, has a relatively low HPV vaccination rate. In this study, we identified factors that impact the intent of Utah parents to vaccinate their children against HPV. A survey was distributed electronically to Utah residents. Survey results were analyzed using confirmatory factor analysis, structural equation modeling, and univariate analysis. Knowledge about HPV and positive vaccine attitudes had the greatest positive effect on intent to vaccinate children against HPV. Cautious sexual attitudes and high religious practice were found to have a negative impact on intent to vaccinate. Effective public health messaging will consider the cultural and religious influences of the target population.

Effects of Religious Practice and Teachings about Sexual Behavior on Intent to Vaccinate against Human Papillomavirus.

Human papillomavirus (HPV) is the most common sexually transmitted infection in the United States. Most infections are mild and clear without treatment in 1 to 2 years. Some HPV strains result in persistent infection, which can cause various cancers, including cervical, penile, anal, mouth, and throat cancers. Vaccines have been developed that provide protection against the highest risk HPV strains. Despite HPV vaccines having been proven to be safe and effective, uptake has been low. Religiosity has been negatively correlated with HPV vaccine uptake in some studies. It is hypothesized that religiosity and Christian religious affiliation could impact parents' decision to vaccinate their children against HPV via teachings and beliefs about sexual behaviors. A survey was distributed to participants to determine what factors, including religiosity and views about sex, impacted HPV vaccination. The survey results (n = 442) were analyzed using confirmatory factor analysis, structural equation modeling, and univariate factor analysis. The association between religious practice and vaccine attitudes were complex, with religious practice slightly positively correlated with pro-vaccine attitudes and vaccine knowledge, but also with the belief that religious adherence to expectations surrounding sexual behavior will protect children from HPV infection, as well as more negative views towards vaccines, in general.