Biological variability of cell-free DNA in healthy females at rest within a short time course.

INTRODUCTION: Alterations in cell-free DNA concentration (cfDNA) over time have been studied in diseased or injured patients or analyzed in athletes during exhaustive exercise. However, no fluctuations have been examined over a short time course in healthy humans at rest so far, wherefore the aim of this study was to examine individual variations at different time points within 75 min. METHODS: Serial blood drawing was performed in 14 healthy female volunteers at rest within 75 min. Plasma DNA was quantified by real-time qPCR, and absolute levels were analyzed together with relative variations. cfDNA alterations were moreover analyzed in consideration of potential volunteer-related impact factors (e.g., pulse) and were compared to alterations of plasma CK and AST. RESULTS: Absolute cfDNA concentration ranged from 0.6 to 3.4 ng/ml. Regarding alterations over time, positive and negative variations were identified, whereby the interdecile range of fold changes was from 0.5 to 1.4. The maximum fold change was determined at 10 min. No relations were found between cfDNA levels and the analyzed individual factors. CONCLUSION: We evidenced the variability of cfDNA in healthy humans at rest within a short time course. The determined variations should serve in future studies to distinguish small cfDNA increases after minor trauma from natural fluctuations. Without such reference of intra-individual variation at rest, it would not be feasible to distinguish an injury from a fluctuation with certainty. Thus, a basis was established for the application of cfDNA as biomarker for the detection of mild injuries in forensic biomechanics.

Perceptions of companion dog benefits on well-being of US military veterans with HIV/AIDS.

OBJECTIVES: Patients with human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) frequently experience psychosocial complications in addition to physical illness. Conflicting data on the value of companion dog ownership in minimizing psychosocial distress suggest the need for more research in this field. This study helps to clarify and expand upon previous research on perceived well-being among patients with HIV/AIDS, specifically as it relates to how owning dogs influences the well-being of US military veterans living with HIV/AIDS. METHODS: Twenty-nine male veterans with a mean age of 52 years who reported having owned a dog since being diagnosed as having HIV/AIDS completed semistructured interviews regarding pet ownership and perceived well-being. Participants also completed a brief survey describing their pets and rating scales that assessed symptoms of depression (nine-question Patient Health Questionnaire-9) and the extent of attachment to their pets (Lexington Attachment to Pets Scale). Descriptive statistics were completed and interview responses were transcribed and examined qualitatively for key themes. RESULTS: The mean Patient Health Questionnaire-9 score of 8.9 (median score of 6) was consistent with mild depressive symptoms, and the mean Lexington Attachment to Pets Scale score was 83.2, indicative of high attachment to one's dog. Veterans reported walking their dogs a mean of 49 minutes/day. Qualitative analysis of the interviews showed that having HIV/AIDS interfered with well-being in three main ways (emotional burden, physical condition, and social isolation). Owning dogs enhanced perceived well-being in four ways (physical activity, companionship, responsibility, and stress reduction). CONCLUSIONS: Twenty-eight of the 29 participants (97%) reported that owning dogs was a positive experience. Overall, this study suggests that veterans with HIV/AIDS who own companion dogs believe that it improves their well-being.

Omicron BA.2 breakthrough infection enhances cross-neutralization of BA.2.12.1 and BA.4/BA.5.

BNT162b2-vaccinated individuals after Omicron BA.1 breakthrough infection have strong serum-neutralizing activity against Omicron BA.1, BA.2, and previous SARS-CoV-2 variants of concern (VOCs) yet less against the highly contagious Omicron sublineages BA.4 and BA.5 that have displaced previous variants. Because the latter sublineages are derived from Omicron BA.2, we characterized serum-neutralizing activity of COVID-19 mRNA vaccine triple-immunized individuals who experienced BA.2 breakthrough infection. We demonstrate that sera of these individuals have broadly neutralizing activity against previous VOCs and all tested Omicron sublineages, including BA.2-derived variants BA.2.12.1 and BA.4/BA.5. Furthermore, applying antibody depletion, we showed that neutralization of BA.2 and BA.4/BA.5 sublineages by BA.2 convalescent sera is driven to a considerable extent by antibodies targeting the N-terminal domain (NTD) of the spike glycoprotein. However, neutralization by Omicron BA.1 convalescent sera depends exclusively on antibodies targeting the receptor binding domain (RBD). These findings suggest that exposure to Omicron BA.2, in contrast to BA.1 spike glycoprotein, triggers substantial NTD-specific recall responses in vaccinated individuals and thereby enhances the neutralization of BA.4/BA.5 sublineages. Given the current epidemiology with a predominance of BA.2-derived sublineages such as BA.4/BA.5 and rapidly ongoing evolution, these findings helped to inform development of our Omicron BA.4/BA.5-adapted vaccine.

Omicron BA.1 breakthrough infection drives cross-variant neutralization and memory B cell formation against conserved epitopes.

Omicron is the evolutionarily most distinct severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variant of concern (VOC) to date. We report that Omicron BA.1 breakthrough infection in BNT162b2-vaccinated individuals resulted in strong neutralizing activity against Omicron BA.1, BA.2, and previous SARS-CoV-2 VOCs but not against the Omicron sublineages BA.4 and BA.5. BA.1 breakthrough infection induced a robust recall response, primarily expanding memory B (BMEM) cells against epitopes shared broadly among variants, rather than inducing BA.1-specific B cells. The vaccination-imprinted BMEM cell pool had sufficient plasticity to be remodeled by heterologous SARS-CoV-2 spike glycoprotein exposure. Whereas selective amplification of BMEM cells recognizing shared epitopes allows for effective neutralization of most variants that evade previously established immunity, susceptibility to escape by variants that acquire alterations at hitherto conserved sites may be heightened.

Exposure to BA.4/5 S protein drives neutralization of Omicron BA.1, BA.2, BA.2.12.1, and BA.4/5 in vaccine-experienced humans and mice.

The SARS-CoV-2 Omicron variant and its sublineages show pronounced viral escape from neutralizing antibodies elicited by vaccination or prior SARS-CoV-2 variant infection owing to over 30-amino acid alterations within the spike (S) glycoprotein. Breakthrough infection of vaccinated individuals with Omicron sublineages BA.1 and BA.2 is associated with distinct patterns of cross-neutralizing activity against SARS-CoV-2 variants of concern (VOCs). In continuation of our previous work, we characterized the effect of Omicron BA.4/BA.5 S glycoprotein exposure on the neutralizing antibody response upon breakthrough infection in vaccinated individuals and upon variant-adapted booster vaccination in mice. We found that immune sera from triple mRNA-vaccinated individuals with subsequent breakthrough infection during the Omicron BA.4/BA.5 wave showed cross-neutralizing activity against previous Omicron variants BA.1, BA.2, BA.2.12.1, and BA.4/BA.5 itself. Administration of a prototypic BA.4/BA.5-adapted mRNA booster vaccine to mice after SARS-CoV-2 wild-type strain-based primary immunization is associated with broader cross-neutralizing activity than a BA.1-adapted booster. Whereas the Omicron BA.1-adapted mRNA vaccine in a bivalent format (wild-type + BA.1) broadens cross-neutralizing activity relative to the BA.1 monovalent booster, cross-neutralization of BA.2 and descendants is more effective in mice boosted with a bivalent wild-type + BA.4/BA.5 vaccine. In naïve mice, primary immunization with the bivalent wild-type + Omicron BA.4/BA.5 vaccine induces strong cross-neutralizing activity against Omicron VOCs and previous variants. These findings suggest that, when administered as boosters, mono- and bivalent Omicron BA.4/BA.5-adapted vaccines enhance neutralization breadth and that the bivalent version also has the potential to confer protection to individuals with no preexisting immunity against SARS-CoV-2.

Are Proinflammatory Cytokines Relevant for the Diagnosis of Prostate Cancer?

BACKGROUND/AIM: Inflammation plays an important role in prostate cancer (PCa). We evaluated proinflammatory cytokines regarding differential diagnosis of PCa in men with PSA levels between 2-10 ng/ml. PATIENTS AND METHODS: Serum samples of 79 men (PSA 2-10 ng/ml) were analyzed for 10 proinflammatory cytokines (IL-6, IL-8, TNF-α, IFN-γ, IL-10, IL-1ß, IL-2, IL-4, IL-12p70, IL-13) and results were evaluated regarding presence of PCa and disease severity. RESULTS: Significant differences between PCa patients and controls were found for IL-6 (p=0.002), IL-8 (p=0.030), and TNF-α (p=0.009), although they were not predictors of PCa in a logistic regression analysis. In addition, IL-6 and TNF-α levels were significantly higher in patients with high-risk PCa (p<0.05). No significant differences were observed regarding the other cytokines. CONCLUSION: In patients with PSA levels between 2-10 ng/ml, IL-6, IL-8, and TNF-α are associated with PCa, and IL-6 and TNF-α are associated with high-risk PCa.

Relevance of Circulating Nucleosomes, HMGB1 and sRAGE for Prostate Cancer Diagnosis.

BACKGROUND/AIM: Evasion from cell death occurs in prostate cancer (PCa). We verified whether serum levels of cell death markers can have diagnostic value in PCa. PATIENTS AND METHODS: A total of 233 men scheduled for prostate biopsy [prostate specific antigen (PSA) level: 2-10 ng/ml] were enrolled. Serum nucleosomes, nucleosomes containing the H3 histone (H3), high mobility group box 1 (HMGB1), and soluble receptor for advanced glycation end products (sRAGE) were analyzed by enzyme immunoassays. RESULTS: There were no differences (p>0.05) in nucleosomes, H3, and sRAGE levels between patients with and without PCa or clinically significant PCa (csPCa). HMGB1 had lower levels in PCa patients (p=0.023) and was a predictor of PCa (p=0.047), but not of csPCa (p=0.180). CONCLUSION: In patients with critical PSA levels between 2-10 ng/ml, HMGB1 had some diagnostic value for overall PCa detection, but it was not predictive of csPCa. Nucleosomes, H3 and sRAGE did not discriminate between PCa or csPCa and controls.