NK cells modulate in vivo control of SARS-CoV-2 replication and suppression of lung damage.

Natural killer (NK) cells play a critical role in virus control. However, it has remained largely unclear whether NK cell mobilization in SARS-CoV-2 infections is beneficial or pathologic. To address this deficit, we employed a validated experimental NK cell depletion non-human primate (NHP) model with SARS-CoV-2 Delta variant B.1.617.2 challenge. Viral loads (VL), NK cell numbers, activation, proliferation, and functional measures were evaluated in blood and tissues. In non-depleted (control) animals, infection rapidly induced NK cell expansion, activation, and increased tissue trafficking associated with VL. Strikingly, we report that experimental NK cell depletion leads to higher VL, longer duration of viral shedding, significantly increased levels of pro-inflammatory cytokines in the lungs, and overt lung damage. Overall, we find the first significant and conclusive evidence for NK cell-mediated control of SARS-CoV-2 virus replication and disease pathology. These data indicate that adjunct therapies for infection could largely benefit from NK cell-targeted approaches.

Natural killer-like B cells are a distinct but infrequent innate immune cell subset modulated by SIV infection of rhesus macaques.

Natural killer-like B (NKB) cells are unique innate immune cells expressing both natural killer (NK) and B cell receptors. As first responders to infection, they secrete IL-18 to induce a critical cascade of innate and adaptive immune cell infiltration and activation. However, limited research exists on the role of NKB cells in homeostasis and infection, largely due to incomplete and erroneous evaluations. To fill this knowledge gap, we investigated the expression of signaling and trafficking proteins, and the in situ localization and transcriptome of naïve NKB cells compared to conventionally-defined NK and B cells, as well as modulations of these cells in SIV infection. Intracellular signaling proteins and trafficking markers were expressed differentially on naïve NKB cells, with high expression of CD62L and Syk, and low expression of CD69, α4ß7, FcRg, Zap70, and CD3z, findings which were more similar to B cells than NK cells. CD20+NKG2a/c+ NKB cells were identified in spleen, mesenteric lymph nodes (MLN), colon, jejunum, and liver of naïve rhesus macaques (RM) via tissue imaging, with NKB cell counts concentrated in spleen and MLN. For the first time, single cell RNA sequencing (scRNAseq), including B cell receptor (BCR) sequencing, of sorted NKB cells confirmed that NKB cells are unique. Transcriptomic analysis of naïve splenic NKB cells by scRNAseq showed that NKB cells undergo somatic hypermutation and express Ig receptors, similar to B cells. While only 15% of sorted NKB cells showed transcript expression of both KLRC1 (NKG2A) and MS4A1 (CD20) genes, only 5% of cells expressed KLRC1, MS4A1, and IgH/IgL transcripts. We observed expanded NKB frequencies in RM gut and buccal mucosa as early as 14 and 35 days post-SIV infection, respectively. Further, mucosal and peripheral NKB cells were associated with colorectal cytokine milieu and oral microbiome changes, respectively. Our studies indicate that NKB cells gated on CD3-CD14-CD20+NKG2A/C+ cells were inclusive of transcriptomically conventional B and NK cells in addition to true NKB cells, confounding accurate phenotyping and frequency recordings that could only be resolved using genomic techniques. Although NKB cells were clearly elevated during SIV infection and associated with inflammatory changes during infection, further interrogation is necessary to acurately identify the true phenotype and significance of NKB cells in infection and inflammation.

Multiplex interrogation of the NK cell signalome reveals global downregulation of CD16 signaling during lentivirus infection through an IL-18/ADAM17-dependent mechanism.

Despite their importance, natural killer (NK) cell responses are frequently dysfunctional during human immunodeficiency virus-1 (HIV-1) and simian immunodeficiency virus (SIV) infections, even irrespective of antiretroviral therapies, with poorly understood underlying mechanisms. NK cell surface receptor modulation in lentivirus infection has been extensively studied, but a deeper interrogation of complex cell signaling is mostly absent, largely due to the absence of any comprehensive NK cell signaling assay. To fill this knowledge gap, we developed a novel multiplex signaling analysis to broadly assess NK cell signaling. Using this assay, we elucidated that NK cells exhibit global signaling reduction from CD16 both in people living with HIV-1 (PLWH) and SIV-infected rhesus macaques. Intriguingly, antiretroviral treatment did not fully restore diminished CD16 signaling in NK cells from PLWH. As a putative mechanism, we demonstrated that NK cells increased surface ADAM17 expression via elevated plasma IL-18 levels during HIV-1 infection, which in turn reduced surface CD16 downregulation. We also illustrated that CD16 expression and signaling can be restored by ADAM17 perturbation. In summary, our multiplex NK cell signaling analysis delineated unique NK cell signaling perturbations specific to lentiviral infections, resulting in their dysfunction. Our analysis also provides mechanisms that will inform the restoration of dysregulated NK cell functions, offering potential insights for the development of new NK cell-based immunotherapeutics for HIV-1 disease.

Multiplex Analysis of Cytokines and Chemokines in Persons Aging With or Without HIV.

People with HIV (PWH) on combination antiretroviral therapy (cART) are living longer lives due to modern cART advances and increased routine medical care. The full landscape of aging with HIV is unclear; given that HIV emerged relatively recently in human history and initially had a high mortality rate, there has not been a substantially aged population to evaluate. In this study, we set out to perform high-throughput plasma analyte profiling by multiplex analysis, focusing on various T helper (Th)-related cytokines, chemokines, and proinflammatory and anti-inflammatory cytokines. The primary goals being to provide reference ranges of these analytes for aging PWH cohorts, as well as testing the utility of high-throughput multiplex plasma assays. The cohort used in this study comprised age-matched healthy donors (32.6-73.5 years of age), PWH on cART (26.7-60.2 years of age), and viremic PWH (27.5-59.4 years of age). The patients in each group were then stratified across the age span to examine age-related impacts of these plasma biomarkers. Our results largely indicate feasibility of plasma analyte monitoring by multiplex and demonstrate a high degree of person-to-person variability regardless of age and HIV status. Nonetheless, we find multiple associations with age, duration of known infection, and viral load, all of which appear to be driven by either prolonged HIV disease progression or long-term use of cART.

Multiplex analysis of cytokines and chemokines in persons aging with or without HIV.

People with HIV (PWH) on combined antiretroviral therapy (cART) are living longer lives due to modern cART advances and increased routine medical care. The full landscape of aging with HIV is unclear; given that HIV emerged relatively recently in human history and initially had a high mortality rate, there has not been a substantially aged population to evaluate. In the present study, we set out to perform high throughput plasma analyte profiling by multiplex analysis, focusing on various T helper (Th)-related cytokines, chemokines, and pro- and anti-inflammatory cytokines. The primary goals being to provide reference ranges of these analytes for aging PWH cohorts, as well as testing the utility of high throughput multiplex plasma assays. The cohort used in this study was comprised of age-matched healthy donors (aged 32.6-73.5), PWH on cART (aged 26.7-60.2), and viremic PWH (aged 27.5-59.4). The patients in each group were then stratified across the age span to examine age-related impacts of these plasma biomarkers. Our results largely indicate feasibility of plasma analyte monitoring by multiplex and demonstrate a high degree of person-to-person variability regardless of age and HIV status. Nonetheless, we find multiple associations with age, duration of known infection, and viral load, all of which appear to be driven by either prolonged HIV disease progression or long-term use of cART.

Child Sexual Grooming of Catholic Clergy: A Comparison between Victims Allegedly Abused by Clergy with Single versus Multiple Victims.

There is evidence that clergy who commit child sexual abuse (CSA) may utilize sexual grooming behaviors in the offense process consistent with a content-validated model (Sexual Grooming Model; SGM). Although research has examined differences in characteristics between clergy with single versus multiple CSA victims, little is known about how sexual grooming behaviors may vary between these groups. The present study utilized a large sample of alleged clergy sexual abuse incidents (n = 10,667) to examine the differences in reported sexual grooming behaviors for victims who experienced abuse by an individual who had a single (V-SVOs) or multiple (V-MVOs) victims. As part of a larger study, files of victim reports of CSA were reviewed from 195 dioceses/eparchies and 140 religious institutes in the United States; these data were analyzed based on behaviors in the SGM for the present study. Overall, the findings showed that the two groups (V-SVOs and V-MVOs) experienced similar tactics throughout the sexual grooming process outlined by the SGM. However, there were some differences for certain tactics within the gaining access and isolation, trust development, and desensitization to sexual contact and physical touch stages of sexual grooming. The implications for prevention, policy, and treatment, as well as future directions of research, are discussed.

Natural Killer Cells Regulate Acute SIV Replication, Dissemination, and Inflammation, but Do Not Impact Independent Transmission Events.

Natural killer (NK) cells are potent effector cells of the innate immune system possessing both cytotoxic and immunoregulatory capabilities, which contribute to their crucial role in controlling human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) infections. However, despite significant evidence for NK cell modulation of HIV disease, their specific contribution to transmission and control of acute infection remains less clear. To elucidate the contribution of NK cells during acute SIV infection, we performed an acute necropsy study, where rhesus macaques (RM) were subjected to preinfection depletion of systemic NK cells using established methods of IL-15 neutralization, followed by subsequent challenge with barcoded SIVmac239X. Our study showed that depletion was highly effective, resulting in near total ablation of all NK cell subsets in blood, liver, oral, and rectal mucosae, and lymph nodes (LN) that persisted through the duration of the study. Meanwhile, frequencies and phenotypes of T cells remained virtually unchanged, indicating that our method of NK cell depletion had minimal off-target effects. Importantly, NK cell-depleted RM demonstrated an early and sustained 1 to 2 log increase in viremia over controls, but sequence analysis suggested no difference in the number of independent transmission events. Acute bulk, central memory (CM), and CCR5+ CD4+ T cell depletion was similar between experimental and control groups, while CD8+ T cell activation was higher in NK cell-depleted RM as measured by Ki67 and PD-1 expression. Using 27-plex Luminex analyses, we also found modestly increased inflammatory cytokines in NK cell-depleted RM compared to control animals. In the effort to determine the impact of NK cells on HIV/SIV transmission and acute viremia, future studies will be necessary to better harness these cells for future viral therapies. Collectively, these data suggest NK cells are important modulators of lentivirus dissemination and disease but may not have the capacity to independently eliminate individual transmission events. IMPORTANCE Natural killer (NK) cells as major effector cells of the innate immune system can contribute significantly to human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) control. However, a specific role for NK cells in blocking lentivirus transmission remains incompletely clear. In this study, we depleted NK cells prior to challenge with a barcoded SIV. Importantly, our studied showed systemic NK cell depletion was associated with a significant increase in acute viremia, but did not impact the number of independent transmission events. Collectively, these data suggest NK cells are critical modulators of early lentivirus replication but may not regulate individual transmission events at mucosal portals of entry.

The Prevalence of Sexual Grooming Behaviors in a Large Sample of Clergy.

Following the exposure of child sexual abuse (CSA) within the U.S. Catholic Church, it was postulated that many of these cases went unreported because sexual grooming was involved. The present study examined the prevalence of sexual grooming behaviors of clergy through the lens of the content-validated Sexual Grooming Model (SGM). The study used archival data from a victim survey of 10,667 cases of alleged CSA within the U.S. Catholic Church. Results revealed the accused clergy frequently used tactics in the gaining access and isolation, trust development, and desensitization stages of the SGM. From the limited data available for the victim selection and post-abuse maintenance stages, the findings showed these tactics were relatively less common. Overall, clergy who were alleged to have committed CSA used tactics consistent with sexual grooming in general, although there were unique aspects of sexual grooming given their institutional role and position.

Prevalence of Journal Clubs: A Survey of Clinical Pastoral Education Residencies in the United States.

Journal clubs are an established and effective method of promoting research literacy and evidence-based practice in the medical field. However, their use in clinical pastoral education (CPE) residencies is relatively new and largely unstudied. In 2016 we surveyed 201 ACPE certified educators of CPE residency programs throughout the United States. Eighty-eight certified educators participated in this first-of-its-kind study. Our aim was to determine the prevalence and structure of existing CPE journal clubs and establish baseline data to determine the effectiveness of future programs for increasing research literacy. Seventy-three percent of respondents indicated they did not have a journal club. Journal clubs that do exist tend to meet at least monthly and last one to two hours. Chaplains, certified educators, or chaplain residents lead the majority of journal clubs. Seventy percent of respondents whose programs had journal clubs indicated they were interested to learn more about journal clubs. Journal clubs do not appear to be common in CPE residencies and chaplains may feel unprepared to lead them. CPE journal clubs would likely be strengthened by collaboration with other disciplines and from additional training for CPE certified educators. We offer suggestions to this end as well as areas for further research.

Child Sexual Abuse in Organizational Settings: A Research Framework to Advance Policy and Practice.

Recent decades have produced growing public attention to the problem of child sexual abuse (CSA) occurring within civic institutions, school settings, youth sports, religious institutions, and other youth-serving organizations. Often amid considerable media and public scrutiny, such institutions have been called upon to improve their responses to sexual abuse incidents, address underlying organizational conditions that may foster such incidents, and develop viable systems of early intervention and prevention. These system challenges, in turn, have produced growing demand for high-quality research that can refine our understanding of the correlates and dynamics of sexual abuse within institutional settings and can help identify the parameters of effective strategies and responses. This article introduces a special issue of Sexual Abuse featuring emerging scholarship focused on CSA within organizational settings and sets forth a framework for further advancing the base of knowledge in this vital area of policy and practice.

Campus Sexual Assault: Forging an Action-Focused Research Agenda.

In recent years, the problem of campus-based sexual assault has emerged as a prominent matter of concern for institutions of higher education. Amid expanded media attention and a shifting policy landscape, many institutions have grappled with programmatic and legal challenges related to systems of investigation and adjudication of sexual assault cases. Meanwhile, many have worked to develop and deploy proactive preventive measures including those involving bystander engagement, peer-driven interventions, sexual assault awareness campaigns, self-defense programs, and the innovative use of social media and other technology. In this context, there is a growing need for high-quality empirical research that can shed light on the extent and nature of campus sexual assault; evaluate existing institutional systems and processes; and promote the development, testing, and evaluation of novel approaches that respond to unmet needs and challenges. As an introduction to a special journal issue devoted to this nascent but rapidly emerging field of inquiry, this article offers context and perspective on the vital role that research can play in the development and advancement of effective policies and strategies to prevent and effectively respond to campus-based sexual assault.

POSTOPERATIVE INSULIN REQUIREMENTS IN BARIATRIC SURGERY.

OBJECTIVE: Though insulin dose reduction months after surgery is a well-studied outcome, there are limited data on immediate postoperative changes. The goals of the present study were to ( 1) To determine peri-operative glycemic control in patients with type 2 diabetes mellitus (DM) on insulin who have undergone Roux-en-Y gastric bypass (RYGB) and ( 2) to compare pre- and postoperative insulin regimens and dosages in these patients. METHODS: A retrospective chart review was conducted on patients with type 2 DM on insulin who underwent RYGB surgery. Blood glucose (BG) levels and insulin doses were compared prior to surgery, on the day of surgery (DOS), and postoperative days (POD) 1 and 2. Subgroup analysis was performed to see if insulin dose was related to glucose control. RESULTS: There were 114 subjects with a mean (SD) age of 52.8 ± 9.8 years, body mass index (BMI) 46.2 ± 8.0 kg/m2, glycated hemoglobin A1c (HbA1c) 8.3% (67 mmol/mol) ± 1.7%, and 66% on insulin plus noninsulin medications and 34% on insulin only. Mean blood glucose (BG) significantly decreased from the DOS (185 ± 43 mg/dL) through POD2 (160 ± 36, P<.0001). The median daily insulin dose significantly decreased from before surgery on usual diet (75 units [36, 116 interquartile range (IQR)]) through POD2 (6 [2, 15 IQR]), P<.0001). The median insulin dose per body weight decreased significantly from before surgery on usual diet (0.58 units/kg [0.35, 0.84 IQR]) through POD2 (0.04 [0.02, 0.11 IQR]), P<.0001). The subgroup with relatively good control experienced a larger percentage reduction in insulin requirements versus subjects with poor control. CONCLUSION: An 87.5% reduction in total daily insulin dose was seen by POD2. This will assist in developing algorithms for insulin titration postbariatric surgery. ABBREVIATIONS: BG = blood glucose DM = diabetes mellitus DOS = day of surgery HbA1c = glycated hemoglobin IQR = interquartile range IV = intravenous NPH = neutral protamine Hagedorn POD = postoperative day RYGB = Roux-en-Y gastric bypass SSRI = sliding scale regular insulin.

Promoter Hypermethylation and Decreased Expression of Syncytin-1 in Pancreatic Adenocarcinomas.

Syncytin-1 is a member of human endogenous retroviral W gene family (HERVW1). Known to be expressed in human placental trophoblast, syncytin-1 protein mediates the fusion of cytotrophoblasts for the formation of syncytiotrophoblasts, the terminally differentiated form of trophoblast lineage. In addition, in vitro studies indicate that syncytin-1 possessed nonfusogenic functions such as those for immune suppression, cell cycle regulation and anti-apoptotic activities. Overexpression of syncytin-1 has been observed in various malignant tissues including breast, endometrial and ovarian cancers. It was reported that syncytin-1 gene expression is associated with dynamic changes of DNA hypomethylation in the 5' LTR. In this study, applying the real-time PCR, Western blot analysis and immunohistochemistry methods, we demonstrate a constitutive expression of syncytin-1 in normal pancreas tissues as well as normal tissues adjacent to cancer lesions. Moreover, a reduced expression is found in the pancreatic adenocarcinoma tissues. The expression levels of syncytin-1 are not correlated with the stage, historical grade and gender, but inversely correlated with patients' age. Furthermore, COBRA and bisulfite sequencing results indicated that the lower expression of syncytin-1 is correlated with the hypermethylation of two CpG dinucleotides in the 5' LTR of syncytin-1 gene. The nonfusogenic function of syncytin-1 in normal pancreas as well as its role(s) in the pathogenesis and progression of pancreatic cancers remains to be investigated. Identification of the two CpG dinucleotides around transcription start site as key epigenetic elements has provided valuable information for further studies on the epigenetic regulation of syncytin-1 in pancreatic cancer cells.

Blessed be the Children: A Case-Control Study of Sexual Abusers in the Catholic Church.

Individuals working in churches and other youth-serving institutions have a unique level of access to children, yet the problem of sexual abuse in institutional settings has received scant research attention. To address this gap, we analyzed data from a large sample of clergy (N = 1,121) and applied a social-ecological model of offending to identify risk factors for sexual abuse perpetration. Using a case-control study design that compared clergy sexual abusers with three control groups of clergy, this study focuses specifically on individual-, relationship-, and community-level factors associated with a higher risk of abuse in professional populations. Findings revealed that clergy sexual abusers tended to have more truncated pre-seminary dating histories, and that their dating and sexual partners were more likely to have been male than female. Self-reported sexual abuse history was associated with a greater likelihood of sexual abuse perpetration among clergy. Clergy abusers tended to be more involved with youth and adolescents in their ministries; however, they were observed to relate less well to youth and adolescents than their clergy counterparts. Given widespread changes in our cultural understanding of abuse as well as more specific changes in the organizational approach to seminary education, these differences underscore the role that youth-serving institutions and society can have in the primary prevention of child sexual abuse.

Expression and diagnostic value of HE4 in pancreatic adenocarcinoma.

Human epididymis protein 4 (HE4) is a recognized biomarker in ovarian and endometrial cancer and over-expressed in pancreatic adenocarcinoma. The diagnostic value of HE4 in pancreatic adenocarcinoma remains unknown. Here we elucidate mRNA, protein and serum level of HE4 in pancreatic adenocarcinoma. HE4 mRNA level in tumor adjacent tissues and pancreatic adenocarcinoma tissues were tested by real time-PCR. Tissue microarray containing normal, adenocarcinoma, and adjacent pancreatic tissue was tested by immunohistochemistry (IHC). Serum level of HE4, carbohydrate antigen 19-9 (CA19-9), carbohydrate antigen 15-3 (CA15-3) and carbohydrate antigen 125 (CA125) were detected by ELISA assay in control and tumor patients. Further we compared the sensitivity and specificity of determining HE4, CA19-9, CA15-3, and CA125 for diagnosis of pancreatic adenocarcinoma and assessed the complementary diagnostic value of HE4, CA19-9, CA15-3 and CA125. Real time PCR showed significantly increased HE4 mRNA level in pancreatic adenocarcinoma compared with control. Result of IHC showed that HE4 significantly higher expressed in the human pancreatic carcinoma tissues than in both normal and adjacent non-tumorous pancreatic tissues, and the staining intensity is inversely correlated with the clinical stage. HE4 was highly expressed in early stage of pancreatic adenocarcinoma. Serum HE4 level is higher in cases with pancreatic adenocarcinoma than in the controls. Serum HE4 levels could research to a sensitivity of 45.83% and specificity of 93.75% when the Cutoff was set at 4.59 ng/mL. The Combined HE4 and CA19-9 increased the sensitivity to 83.33%; and interestingly, the combination of HE4 with CA15-3 led to the most powerful sensitivity of 87.5%. Combined with CA19-9 and CA15-3, HE4 could be a potential biomarker to improve the diagnostic power for pancreatic adenocarcinoma.

Understanding child sexual abuse by Catholic priests from a situational perspective.

Most sexual offense research focuses on offender motivation and individual risk factors rather than the criminal events themselves. This article provides an analysis of data from two studies on child sexual abuse by Catholic priests to help understand the opportunities clergy had or created to abuse youth. Findings show that situational factors played a role in victim choice among a heterogeneous group of abusers. Priests abused more male than female victims and had significantly greater access to male youth historically. When access to female youth increased in the 1990s, abuse of females as a percentage of victims also increased. The article concludes with a discussion about how ecological information can be used to craft intervention policies to prevent sexual offenses.

Development, implementation, and evaluation of a comprehensive fall risk program.

PURPOSE: The purpose was to develop and implement I'M SAFE, a comprehensive patient fall-risk assessment tool linked to a tiered-intervention falls prevention program. DESIGN AND METHOD: A fall-risk evaluation tool was incorporated into electronic nurse documentation along with risk-specific nursing interventions. RESULTS: Intrinsic fall rates declined significantly (preimplementation: .67 falls/1,000 patient days; postimplementation: .51 falls/1,000 patient days, p = .015) and has been sustained 2 years following implementation. PRACTICE IMPLICATIONS: The I'M SAFE tool identifies patients at increased risk for falls. When linked to a multidisciplinary fall prevention program, the incidence of preventable falls can be reduced. The program's impact has persisted across two facilities.