NKG2D receptor signaling shapes T cell thymic education.

The role of natural killer group 2D (NKG2D) in peripheral T cells as a costimulatory receptor is well established. However, its contribution to T cell thymic education and functional imprint is unknown. Here, we report significant changes in development, receptor signaling, transcriptional program, and function in T cells from mice lacking NKG2D signaling. In C57BL/6 (B6) and OT-I mice, we found that NKG2D deficiency results in Vß chain usage changes and stagnation of the double-positive stage in thymic T cell development. We found that the expression of CD5 and CD45 in thymocytes from NKG2D deficient mice were reduced, indicating a direct influence of NKG2D on the strength of T cell receptor (TCR) signaling during the developmental stage of T cells. Depicting the functional consequences of NKG2D, peripheral OT-I NKG2D-deficient cells were unresponsive to ovalbumin peptide stimulation. Paradoxically, while αCD3/CD28 agonist antibodies led to phenotypic T cell activation, their ability to produce cytokines remained severely compromised. We found that OT-I NKG2D-deficient cells activate STAT5 in response to interleukin-15 but were unable to phosphorylate ERK or S6 upon TCR engagement, underpinning a defect in TCR signaling. Finally, we showed that NKG2D is expressed in mouse and human thymic T cells at the double-negative stage, suggesting an evolutionarily conserved function during T cell development. The data presented in this study indicate that NKG2D impacts thymic T cell development at a fundamental level by reducing the TCR threshold and affecting the functional imprint of the thymic progeny. In summary, understanding the impact of NKG2D on thymic T cell development and TCR signaling contributes to our knowledge of immune system regulation, immune dysregulation, and the design of immunotherapies.

Longitudinal findings of a U.S. preventive evidence-based family intervention tested among youth in Ecuador: Familias Unidas.

Interventions that address adolescent conduct problems are essential for decreasing negative risk behaviors and promoting positive protective factors among youth. Although interventions have been developed and tested in the United States, preventive evidence-based interventions (EBIs) are less available in Latin American countries such as Ecuador. Therefore, the purpose of this study was to evaluate the efficacy of an evidence-based, parent-centered intervention, Familias Unidas, in preventing/reducing conduct problems, across time, among youth in Guayaquil, Ecuador. Ecuadorian youth (ages 12 through 14) and their respective primary caregiver were recruited from two public schools and randomized to either Familias Unidas or Community Practice. A series of latent growth models were run to test for differences between Familias Unidas and Community Practice on conduct disorder symptoms across three timepoints covering 6 months. Ecuadorian mental health professionals were trained to deliver the evidence-based intervention. Findings indicate no direct relationship between condition and average change in conduct problems at 6 months post baseline. However, indirect effects favoring Familias Unidas over Community Practice were found through improvements in family functioning. Findings highlight that Familias Unidas was efficacious in an international setting and indicate the viability of successfully delivering preventive EBIs in Ecuador.

Memory Precursors and Short-Lived Effector T cell Subsets Have Different Sensitivities to TGFß.

After exposure to an antigen, CD8 T cells reach a decision point about their fate: to become either short-lived effector cells (SLECs) or memory progenitor effector cells (MPECs). SLECs are specialized in providing an immediate effector function but have a shorter lifespan and lower proliferative capacity compared to MPECs. Upon encountering the cognate antigen during an infection, CD8 T cells rapidly expand and then contract to a level that is maintained for the memory phase after the peak of the response. Studies have shown that the contraction phase is mediated by TGFß and selectively targets SLECs, while sparing MPECs. The aim of this study is to investigate how the CD8 T cell precursor stage determines TGFß sensitivity. Our results demonstrate that MPECs and SLECs have differential responses to TGFß, with SLECs being more sensitive to TGFß than MPECs. This difference in sensitivity is associated with the levels of TGFßRI and RGS3, and the SLEC-related transcriptional activator T-bet binding to the TGFßRI promoter may provide a molecular basis for increased TGFß sensitivity in SLECs.

Family Functioning in Hispanic Parents of Adolescents: Who Benefits Most from a Family-Based HIV and Substance Use Preventive Intervention?

To understand which families are likely to benefit most from resource-intensive family-based, evidence-based interventions (EBIs), we must examine the key, modifiable determinant of family functioning. The purpose of this study was to (1) identify whether there are subgroups of Hispanic parents that differ meaningfully based on their family functioning at baseline, (2) test whether the Familias Unidas preventive intervention was differentially effective across the baseline family functioning subgroups, and (3) understand the mechanisms of intervention effectiveness within each baseline family functioning subgroup. On a pooled data set of 4 completed efficacy and effectiveness trials of Familias Unidas (n = 1445 low-income, Hispanic immigrant origin, parents and their adolescent between the ages of 12-17), we conducted a series of secondary data analyses. Latent profile analyses revealed four significantly different profiles: (1) low family functioning (n = 210, 14.55%), (2) low-to-moderate family functioning (n = 554, 38.39%), (3) moderate-to-high family functioning (n = 490, 33.96%), and (4) high family functioning (n = 189, 13.10%). A structural equation modeling approach found there were significant differences in intervention effectiveness between the subgroups. The low family functioning subgroup experienced gains in family functioning, and in turn, lower levels of adolescent substance use, internalizing, and externalizing symptoms. The high family functioning subgroup showed significant direct effects of the intervention on adolescent substance use, internalizing, and externalizing symptoms, but no indirect effects through improvements in family functioning. Implications for screening, targeting, and adapting interventions are discussed.

Metagenomic analysis of the intestinal microbiome in goats on cactus and Salicornia-based diets.

Background: The Peruvian coast is characterized by its arid and saline soils, the cactus being an alternative for arid soils and Salicornia for saline soils. Therefore, it is necessary to develop nutrition based on the intestinal microbiota in goats. Aim: To identify the intestinal microbiota in goats through a metagenomic analysis. Methods: In this study, goats and kids were randomly selected and fed cacti and Salicornia as potential forage species compared to native grass to study the changes in the microbiota using massive sequencing using the 16S rRNA gene as a marker. Results: The sequencing results showed the taxonomic levels of Bacteroidetes and Firmicutes at the phylum level as the most abundant in creole goats' microbiome, varying from 18% to 36% and 47% to 66%, respectively. At the genus level, variants of the genus Ruminococcaceae stand out, related to cellulose degradation, as the most dominant in all samples, followed by Christensenellaceae, Rikenellaceae, and Prevotellaceae. Also, the genus Akkermansia appeared in greater abundance in kids fed with cactus, being necessary for being related to the intestinal mucosa's health and avoiding the adhesion of pathogens to the intestinal epithelium. Conclusion: These microbiota changes based on diets with high fiber content are necessary to understand the adaptation of this species to favorable dietary changes.

Resurrection and redescription of Clepsinepallida Verrill, 1872 (Hirudinida, Glossiphoniidae) with a phylogeny of the genus Alboglossiphonia.

Alboglossiphoniapallida (Verrill, 1872) comb. nov. is resurrected and redescribed based on morphological and molecular data from specimens of the type locality (New Haven County, Connecticut, USA) that demonstrate it is distinct from North American Alboglossiphoniaheteroclita, European Alboglossiphoniaheteroclita, and Alboglossiphoniapapillosa. Alboglossiphoniapallida is characterized by having dark chromatophores on the dorsal surface arranged lateral to patrilaterally and medially as a thin line or interrupted thin line along with three pairs of eye spots (with the first pair closest together), six pairs of crop ceca, and a united gonopore. Additional sampling of specimens of the genus Alboglossiphonia is needed to understand its phylogeny especially as many species have not been collected since their description.

Corrigendum: The Role of NKG2D in Vitiligo.

[This corrects the article DOI: 10.3389/fimmu.2021.624131.].

Socio-Cultural Subgroups of Latina/o Immigrants: A Latent Profile Analysis.

Socio-cultural factors, such as familismo, social support, machismo, and multigroup ethnic identity, are strongly associated with Latina/o immigrants' alcohol misuse and depressive symptoms. However, research has rarely explored whether unobserved groups of Latina/o immigrants with similar socio-cultural factors exist. Latent Profile analysis can illuminate which subgroups to target, or which socio-cultural factors need to be supported, to have an impact on the prevention and treatment of alcohol use and/or depression in the Latina/o immigrant population. Cross sectional data from on ongoing longitudinal investigation was utilized (N= 518 Latina/o adults living in Miami-Dade County, Florida and have immigrated to the US within one year prior to assessment). Latent Profile Analyses (LPA) were conducted utilizing validated measures of familismo, social support, neighborhood collective efficacy, ethnic identity, machismo, caballerismo, and marianismo. The LPA revealed three, significantly different profiles: (1) low socio-cultural protection (n=155, 29.98%), (2) high socio-cultural protection (n=21, 4.06%), and (3) high socio-, low-cultural protection (n=341, 65.96%). Profile membership was associated significantly with immigrant documentation status, education level, and past family history of substance use. Results indicate that Latina/o immigrants in the low socio-cultural protection group had significantly higher alcohol use compared to high socio-, low cultural protection group. No significant differences were found for depressive symptoms. We discuss implications of our findings and encourage researchers to continue to unpack the complexities associated with socio-cultural factors and Latina/o mental and behavioral health. Specifically, research should focus on socio-cultural factors can provide protection from negative health outcomes and increase resiliency among this population.

The Role of the NKG2D in Vitiligo.

Vitiligo is an acquired multifactorial disease that affects melanocytes and results in skin depigmentation. In this review, we examine the role of cells stress and self-reactive T cells responses. Given the canonical and non-canonical functions of NKG2D, such as authenticating stressed target and enhance TCR signaling, we examine how melanocyte stress leads to the expression of ligands that are recognized by the activating receptor NKG2D, and how its signaling results in the turning of T cells against self (melanocyte suicide by proxy). We also discuss how this initiation phase is followed by T cell perpetuation, as NKG2D signaling results in self-sustained long-lasting T cells, with improved cytolytic properties.

A Feasibility Trial of an Online-Only, Family-Centered Preventive Intervention for Hispanics: e-Familias Unidas.

We piloted the preventive intervention e-Familias Unidas, delivered exclusively through the Internet to maximize reach and sustainability. This program is an adaptation of an evidence-based, family-centered intervention that aims to prevent Hispanic adolescent drug use and risky sexual behavior through improving family functioning. The purpose of this feasibility trial was to: (1) explore the use of Facebook and a trusted community champion in the online recruitment of Hispanic parents of adolescents, (2) test an online-only assessment and randomization protocol, (3) pilot intervention delivery via e-familiasunidas.com, and (4) assess pre-post changes in family functioning. We tracked participant recruitment and enrollment via Facebook analytics and REDCap. Intervention completion was tracked via our website. We conducted an ANCOVA to test for between group (e-Familias Unidas vs. control) differences post-intervention. Recruitment lasted for a total of 10 weeks and was divided into three phases, with each phase systematically assessing which strategies maximized recruitment and enrollment. Overall, 93 Hispanic parents enrolled in the study. Of those randomized to the intervention group (n = 46), 70% were engaged and watched an average of 5.4 out of the 12 online videos. We found that participants recruited through a community champion, versus Facebook advertisements, had higher rates of enrollment and intervention completion. There was a significant improvement in parent-adolescent communication for parents who received e-Familias Unidas compared to those in the control group (n = 47). This pilot trial demonstrated it is feasible to recruit, assess, and deliver e-Familias Unidas to Hispanic parents through an online-only platform. Our findings highlight the promise of an online platform to optimize the reach of preventive interventions for underserved populations, to more effectively target participants, and to disseminate sustainable evidence-based interventions. We discuss lessons learned and recommendations for future research.

Parent Attendance in a Family-Based Preventive Intervention Delivered in Latin America and the United States.

In Latin America, there is an increasing interest in the implementation and dissemination of evidence-based, family-centered interventions to prevent youth behavioral problems. While families' participation in interventions is integral to achieving the interventions' desired impact, little is known about what predicts Latin American families' attendance. The current study provides a unique opportunity to explore the participation of families living in the United States, Ecuador, and Chile in an evidence-based intervention, Familias Unidas. We tested for differences in attendance rates, family functioning variables, and adolescent behavioral problem variables, then applied a hierarchical multiple regression to (a) identify which variables significantly predicted program attendance and (b) assess whether the country in which the intervention was implemented in moderated the relationship between predictors and program attendance. On average, Chilean and Ecuadorian parents were more engaged and attended more sessions than parents living in the United States. Across samples, there was significant differences in family functioning and adolescent behavioral problem variables. However, effective parent-adolescent communication was the only significant predictor of lower program attendance. A significant interaction effect revealed that even though Chilean parents had high parent-adolescent communication, they were more likely to attend sessions, compared to parents living in the United States. We highlight the promise of engaging and retaining families, across U.S. and Latin American samples, into a culturally syntonic, family-based intervention, and discuss potential explanations for success in Chile and Ecuador. Researchers interested in implementing interventions in Latin America could utilize these findings to better target participants and intervention efforts.

A facile solvent extraction method facilitating surface-enhanced Raman spectroscopic detection of ochratoxin A in wine and wheat.

The capability of a solvent-mediated liquid-liquid extraction (LLE) method to improve the detection of ochratoxin A (OTA) in food matrixes using surface-enhanced Raman spectroscopy (SERS) is described. SERS detection of mycotoxins with nanoparticle aggregation is a simple method but with low reproducibility due to the heterogeneous distribution of the nanoparticle aggregates. We evaluated three different LLE protocols to analyze their performance in combination with SERS. A facile extraction method based on sample acidification and addition of chloroform as a separation solvent showed to not only extract OTA from wine and wheat but also facilitate the uniform distribution of the nanoparticles leading to an improvement of the detection signals and the reproducibility. This method enables rapid and simple analysis of mycotoxin Ochratoxin A in food systems.

Characterization of Actinobacillus seminis biofilm formation.

Actinobacillus seminis is an autochthonous gram-negative bacterium that affects reproductive organs, causing epididymitis, low fertility, and occasional abortions in ovine and goats. The virulence factors and the pathogenicity mechanisms of A. seminis have not been clearly elucidated yet. In this work, biofilm production by A. seminis in in vitro assays is described and characterized. After 48-h incubation at 37 °C in trypticase soy broth, A. seminis formed biofilms containing an extracellular matrix comprised mainly of fibrillar material. Microaerophilia or the presence of calcium diminished biofilm formation in approximately 50% and 70%, respectively, but low iron concentrations increased it 40%. Through enzymatic digestion, it was found that proteins were the main component of these biofilms. Structural observations through scanning electron microscopy indicated the presence of a high amount of fibrillar material in which bacteria were immersed. Antibodies against different bacterial surface proteins, such as anti-biofilm matrix and anti-adhesin, diminished biofilm formation in 70% and 25%, respectively; whereas furanone C-30 and LED-209, compounds described as quorum-sensing inhibitors, completely inhibited biofilm formation. In conclusion, environmental conditions can influence strongly biofilm formation in A. seminis, and this could be an advantageous strategy that allows bacteria to persist inside a host.

Parenting Interventions in Pediatric Primary Care: A Systematic Review.

CONTEXT: More than 4 decades of research indicate that parenting interventions are effective at preventing and treating mental, emotional, and behavioral disorders in children and adolescents. Pediatric primary care is a viable setting for delivery of these interventions. OBJECTIVE: Previous meta-analyses have shown that behavioral interventions in primary care can improve clinical outcomes, but few reviews have been focused specifically on the implementation of parenting interventions in primary care. We aimed to fill this gap. DATA SOURCES: We reviewed 6532 unique peer-reviewed articles published in PubMed, the Cumulative Index to Nursing and Allied Health Literature, and PsycInfo. STUDY SELECTION: Articles were included if at least part of the intervention was delivered in or through primary care; parenting was targeted; and child-specific mental, emotional, and behavioral health outcomes were reported. DATA EXTRACTION: Articles were reviewed in Covidence by 2 trained coders, with a third coder arbitrating discrepancies. RESULTS: In our review of 40 studies, most studies were coded as a primary. Few researchers collected implementation outcomes, particularly those at the service delivery system level. LIMITATIONS: Including only published articles could have resulted in underrepresentation of implementation-related data. CONCLUSIONS: Parenting interventions delivered and implemented with fidelity in pediatric primary care could result in positive and equitable impacts on mental, emotional, and behavioral health outcomes for both parents and their children. Future research on the implementation strategies that can support adoption and sustained delivery of parenting interventions in primary care is needed if the field is to achieve population-level impact.

Inhibition of Histone Deacetylase (HDAC) Enhances Checkpoint Blockade Efficacy by Rendering Bladder Cancer Cells Visible for T Cell-Mediated Destruction.

Inhibitory checkpoint blockade therapy is an immunomodulatory strategy that results in the restoration of T cell functions, and its efficacy depends on the recognition of tumor cells for destruction. Considering the factors at play, one could propose that anti-tumor responses will not occur if tumor cells are immunologically invisible to T cells. In this study, we tested a strategy based on the modulation of cancer cell's immunovisibility through HDAC inhibition. In a model (heterotopic and orthotopic) of mouse urothelial bladder cancer, we demonstrated that the use of intratumoral or intravesical HDACi in combination with systemic anti-PD-1 was effective at inducing curative responses with durable anti-tumor immunity capable of preventing tumor growth at a distal site. Mechanistically, we determined that protective responses were dependent on CD8 cells, but not NK cells. Of significance, in an in vitro human model, we found that fully activated T cells fail at killing bladder cancer cells unless tumor cells were pretreated with HDACi. Complementary to this observation, we found that HDACi cause gene deregulation, that results in the upregulation of genes responsible for mediating immunorecognition, NKG2D ligands and HSP70. Taken together, these data indicate that HDAC inhibition results in the elimination of the tumor cell's "invisibility cloak" that prevents T cells from recognizing and killing them. Finally, as checkpoint blockade therapy moves into the adjuvant setting, its combined use with locally administrated HDACi represents a new approach to be included in our current therapeutic treatment toolbox.

It Takes Two to Tango.

In the study by Jacquemin et al., the authors reported that ligands for NKG2D are upregulated in vitiligo perilesional skin and especially in patients with active disease. The reasons for the elevated expression of NKG2D ligands are unknown. This study, however, provides a framework for understanding vitiligo: Skin resident CD8 T cells recognize and kill melanocytes through NKG2D signaling. This event results in the increased production and release of cyto/chemokines and the development of long-lasting CD8 T cells, which in turn causes the recruitment of new T cells, thus perpetuating and disseminating the disease.

Enhancing the Routine Screening Infrastructure to Address a Syphilis Epidemic in Miami-Dade County.

BACKGROUND: In a recent sexually transmitted disease surveillance report from the Centers for Disease Control and Prevention, Miami-Dade County had the nation's fourth highest rate of infectious syphilis, with rates of congenital syphilis on the rise. As a public health response, Homestead Hospital, in collaboration with the Florida Department of Health, enhanced their emergency department's routine HIV/HCV "opt-out" screening infrastructure to include a syphilis smart screening algorithm. The purpose of this article is to describe the development, implementation, and evaluation of the algorithm. METHODS: A retrospective evaluation of patient records prompted the development of the algorithm. Homestead Hospital's electronic health record system automatically triggers a syphilis test based on the reason for medical visit (e.g., rash, penile discharge, a positive pregnancy test, historical or present sexually transmitted disease result). If a patient tests positive, he/she is counseled and linked to care. RESULTS: Since implementation (April 2018 to August 2019), the smart screening algorithm triggered 4806 syphilis tests: 122 patients tested positive (2.5% seropositivity). After confirmatory testing, 59 patients were positive for syphilis, of which 27 were pregnant. CONCLUSIONS: The Homestead Hospital and Department of Health-Miami-Dade's response to Miami-Dade County's syphilis problem is innovative and replicable. The program embraces technology, enhances the routine opt-out screening model, and does not affect preexisting workflows. Ultimately, implementation of this algorithm allows patients to get treatment, receive comprehensive prevention services, and, in some cases, avert congenital syphilis.

A developmental cascade perspective of paediatric obesity: A systematic review of preventive interventions from infancy through late adolescence.

The goals of this systematic review were to identify and describe paediatric obesity prevention interventions from infancy to late adolescence and to provide recommendations for future intervention research in light of a recently proposed developmental cascade (DC) model of paediatric obesity. We conducted an electronic search of randomized controlled trials with a minimum 6-month postintervention follow-up published between 1995 and 2019. We included 74 interventions: prenatal/infancy (n = 4), early childhood (n = 11), childhood (n = 38), early to mid-adolescence (n = 18), and late adolescence (n = 3). Infancy and early childhood trials targeted early feeding and positive parenting skills. Half of the childhood and adolescence trials were school based and used universal prevention strategies; those classified as selective or indicated prevention tended to involve the family for more intensive lifestyle modification. Less than 10% of studies followed participants over long periods of time (greater than or equal to 5 years), and only 16% and 31% of studies assessed intervention mediators and moderators, respectively. We recommend that future interventions focus on early prevention, assess long-term intervention effects, use a standardized taxonomy for defining intervention behavioural strategies, assess underlying mechanisms of action and intervention moderators, target parent and family management strategies across development, and increase scientific equity. We also provide specific recommendations regarding intervention targets for each developmental stage.

Eikenella corrodens causing deep-seated infections. Six-year experience in a University Hospital in Madrid.

INTRODUCTION: Eikenella corrodens (EC) is part of the normal microbiota of the oropharynx and a recognised opportunistic pathogen. It is mainly involved in head and neck infections, but it has also been identified as a cause of pleuropulmonary and intraabdominal infections. Its identification could be difficult due to its fastidious growth requirements, especially in the context of polymicrobial infection and is probably underreported. METHODS: We carried out a retrospective 5-year review of clinical charts and laboratory database. RESULTS: We describe the clinical and microbiological characteristics of 9 deep-seated infections caused by EC, diagnosed in locations different from the head and neck. CONCLUSION: EC deep-seated infections are often found in patients with comorbid conditions and a history of interventional procedures. Due to the characteristic torpid evolution of EC abscesses, imaging to assess the necessity of debridement and avoid early cessation of antibiotics is necessary.

NKG2D signaling certifies effector CD8 T cells for memory formation.

BACKGROUND: The development of memory responses is an evolutionary function of the adaptive immune system. We propose that for the immune system to populate the memory compartment with the best-suited CD8 T cells it utilizes a process of certification or molecular accreditation mediated through Natural Killer Group 2D (NKG2D). This process of certification assures that the memory compartment is filled with CD8 T cells that have demonstrated their ability to kill their cognate targets through a two-step process that utilizes T cell receptor (TCR) and NKG2D signaling. METHODS: One week after immunization with peptide-pulsed dendritic cells, NKG2D signaling was transiently blocked in vivo with a single injection of neutralizing antibodies. Under such conditions, we determined the importance of NKG2D signaling during the effector phase for memory formation without compromising NKG2D signaling at the memory phase. Both open (polyclonal) and closed (monoclonal) CD8 T cell repertoires were studied. RESULTS: We show that signaling through NKG2D mediated this certification. Temporary blockade of NKG2D signaling during the effector phase resulted in the formation of highly defective memory CD8 T cells characterized by altered expression of the ribosomal protein S6 and epigenetic modifiers, suggesting modifications in the T cell translational machinery and epigenetic programming. Finally, these uncertified memory cells were not protective against a B16 tumor challenge. CONCLUSION: Signaling through NKG2D during the effector phase (certification) favors the development of functional memory CD8 T cells, a previously undescribed role for NKG2D. Temporary blockade of NKG2D signaling during the effector phase results in the formation of highly defective memory CD8 T cells potentially by affecting the expression of the ribosomal protein S6 and epigenetic modifiers, suggesting alterations in T cell translational machinery and epigenetic programming.