Blockade of LAG-3 and PD-1 leads to co-expression of cytotoxic and exhaustion gene modules in CD8+ T cells to promote antitumor immunity.

Relatlimab (rela; anti-LAG-3) plus nivolumab (nivo; anti-PD-1) is safe and effective for treatment of advanced melanoma. We designed a trial (NCT03743766) where advanced melanoma patients received rela, nivo, or rela+nivo to interrogate the immunologic mechanisms of rela+nivo. Analysis of biospecimens from this ongoing trial demonstrated that rela+nivo led to enhanced capacity for CD8+ T cell receptor signaling and altered CD8+ T cell differentiation, leading to heightened cytotoxicity despite the retention of an exhaustion profile. Co-expression of cytotoxic and exhaustion signatures was driven by PRDM1, BATF, ETV7, and TOX. Effector function was upregulated in clonally expanded CD8+ T cells that emerged after rela+nivo. A rela+nivo intratumoral CD8+ T cell signature was associated with a favorable prognosis. This intratumoral rela+nivo signature was validated in peripheral blood as an elevated frequency of CD38+TIM3+CD8+ T cells. Overall, we demonstrated that cytotoxicity can be enhanced despite the retention of exhaustion signatures, which will inform future therapeutic strategies.

TPP1 promoter mutations cooperate with TERT promoter mutations to lengthen telomeres in melanoma.

Overcoming replicative senescence is an essential step during oncogenesis, and the reactivation of TERT through promoter mutations is a common mechanism. TERT promoter mutations are acquired in about 75% of melanomas but are not sufficient to maintain telomeres, suggesting that additional mutations are required. We identified a cluster of variants in the promoter of ACD encoding the shelterin component TPP1. ACD promoter variants are present in about 5% of cutaneous melanoma and co-occur with TERT promoter mutations. The two most common somatic variants create or modify binding sites for E-twenty-six (ETS) transcription factors, similar to mutations in the TERT promoter. The variants increase the expression of TPP1 and function together with TERT to synergistically lengthen telomeres. Our findings suggest that TPP1 promoter variants collaborate with TERT activation to enhance telomere maintenance and immortalization in melanoma.

Co-expression of TNF receptors 1 and 2 on melanomas facilitates soluble TNF-induced resistance to MAPK pathway inhibitors.

BACKGROUND: The effectiveness of MAPK pathway inhibitors (MAPKi) used to treat patients with BRAF-mutant melanoma is limited by a range of resistance mechanisms, including soluble TNF (solTNF)-mediated NF-kB signaling. solTNF preferentially signals through type-1 TNF receptor (TNFR1), however, it can also bind to TNFR2, a receptor that is primarily expressed on leukocytes. Here, we investigate the TNFR2 expression pattern on human BRAFV600E+ melanomas and its role in solTNF-driven resistance reprogramming to MAPKi. METHODS: Flow cytometry was used to test TNFR1, TNFR2 and CD271 expression on, as well as NF-kB phosphorylation in human BRAF-mutant melanoma. The ability of melanoma cell lines to acquire MAPKi resistance in response to recombinant or macrophage-derived TNF was evaluated using the MTT cytotoxicity assay. Gene editing was implemented to knock out or knock in TNF receptors in melanoma cell lines. Knockout and knock-in cell line variants were employed to assess the intrinsic roles of these receptors in TNF-induced resistance to MAPKi. Multicolor immunofluorescence microscopy was utilized to test TNFR2 expression by melanoma in patients receiving MAPKi therapy. RESULTS: TNFR1 and TNFR2 are co-expressed at various levels on 4/7 BRAFV600E+ melanoma cell lines evaluated in this study. In vitro treatments with solTNF induce MAPKi resistance solely in TNFR2-expressing BRAFV600E+ melanoma cell lines. TNFR1 and TNFR2 knockout and knock-in studies indicate that solTNF-mediated MAPKi resistance in BRAFV600E+ melanomas is predicated on TNFR1 and TNFR2 co-expression, where TNFR1 is the central mediator of NF-kB signaling, while TNFR2 plays an auxiliary role. solTNF-mediated effects are transient and can be abrogated with biologics. Evaluation of patient specimens indicates that TNFR2 is expressed on 50% of primary BRAFV600E+ melanoma cells and that MAPKi therapy may lead to the enrichment of TNFR2-expressing tumor cells. CONCLUSIONS: Our data suggest that TNFR2 is essential to solTNF-induced MAPKi resistance and a possible biomarker to identify melanoma patients that can benefit from solTNF-targeting therapies.

Illegal Wildlife Trade and Emerging Infectious Diseases: Pervasive Impacts to Species, Ecosystems and Human Health.

Emerging infectious disease (EID) events can be traced to anthropogenic factors, including the movement of wildlife through legal and illegal trade. This paper focuses on the link between illegal wildlife trade (IWT) and infectious disease pathogens. A literature review through Web of Science and relevant conference proceedings from 1990 to 2020 resulted in documenting 82 papers and 240 identified pathogen cases. Over 60% of the findings referred to pathogens with known zoonotic potential and five cases directly referenced zoonotic spillover events. The diversity of pathogens by taxa included 44 different pathogens in birds, 47 in mammals, 16 in reptiles, two in amphibians, two in fish, and one in invertebrates. This is the highest diversity of pathogen types in reported literature related to IWT. However, it is likely not a fully representative sample due to needed augmentation of surveillance and monitoring of IWT and more frequent pathogen testing on recovered shipments. The emergence of infectious disease through human globalization has resulted in several pandemics in the last decade including SARS, MERS, avian influenza H1N1,and Ebola. We detailed the growing body of literature on this topic since 2008 and highlight the need to detect, document, and prevent spillovers from high-risk human activities, such as IWT.

Salmonella serovars associated with Grenadian tree boa (Corallus grenadensis) and their antimicrobial susceptibility.

Cloacal swabs from 45 Grenada bank tree boas (Corallus grenadensis) were sampled during a 12-month period (2011-2012) from the rain forests and scrublands of Grenada. Cloacal swabs were examined by enrichment and selective culture for the presence of Salmonella spp. In all, 16 (35.6%) of the snakes were positive for Salmonella, and six serovars of Salmonella were isolated. The most common serovar was Rubislaw (31.3%), the most frequent serovar recently isolated from green iguanas in Grenada, followed by serovar Braenderup (18.8%), and serovar IV:48:g,z51:- (formerly, S. Marina) (18.8%), also found in green iguanas in this country. The remaining three less frequent serovars were, IV:53:g,z51:-, I:6,7:e,h:- and IIIb:38:i:z. Antimicrobial susceptibility tests conducted by a disc diffusion method against amoxicillin-clavulanic acid, ampicillin, cefotaxime, ciprofloxacin, enrofloxacin, gentamicin, imipenem, nalidixic acid, streptomycin, tetracycline and trimethoprim-sulfamethoxazole showed that drug resistance is minimal, with intermediate susceptibility, only to streptomycin. This is the first report of isolation and antimicrobial susceptibilities of Salmonella serovars from wild Grenadian tree boas.

The Effects of the Putative Confession and Parent Suggestion on Children's Disclosure of a Minor Transgression.

PURPOSE: This study examined the effects of the putative confession (telling the child that an adult "told me everything that happened and he wants you to tell the truth") on children's disclosure of a minor transgression after questioning by their parents. METHODS: Children (N = 188; 4 - 7-year-olds) played with a confederate, and while doing so, for half of the children, toys broke. Parents then questioned their children about what occurred, and half of the parents were given additional scripted suggestive questions. Finally, children completed a mock forensic investigative interview. RESULTS: Children given the putative confession were 1.6 times more likely in free recall to disclose truthfully that toys had broken. Among children who failed to disclose during free recall, those who received the putative confession were 1.9 times more likely when asked yes/no questions to disclose true breakage. The putative confession did not decrease accuracy, and children who received the putative confession were 2.6 times less likely to report false toy play. Parent suggestion did not adversely affect the efficacy of the putative confession. CONCLUSIONS: The current study demonstrates that children are often quite reticent to disclose transgressions, and that the putative confession is a promising avenue for increasing children's comfort with disclosing and minimizing their tendency to report false details, even in the face of suggestive questioning by parents.

The leukemic oncoprotein NPM1-RARA inhibits TP53 activity.

The variant acute promyelocytic leukemia (APL) translocation t(5;17)(q35;q21) fuses the N-terminus of nucleophosmin (NPM1) to the retinoic acid receptor alpha (RARA). We found that ectopic NPM1-RARA expression decreased TP53 protein levels in target cells. NPM1-RARA impaired TP53-dependent transcription. Cells expressing NPM1-RARA were more resistant to apoptotic stimuli. This work identifies the TP53 tumor suppressor as a novel target through which NPM1-RARA impacts leukemogenesis, and confirms the importance of impairment of TP53 in establishment of the APL phenotype.

Stress and emotional valence effects on children's versus adolescents' true and false memory.

Despite considerable interest in understanding how stress influences memory accuracy and errors, particularly in children, methodological limitations have made it difficult to examine the effects of stress independent of the effects of the emotional valence of to-be-remembered information in developmental populations. In this study, we manipulated stress levels in 7-8- and 12-14-year-olds and then exposed them to negative, neutral, and positive word lists. Shortly afterward, we tested their recognition memory for the words and false memory for non-presented but related words. Adolescents in the high-stress condition were more accurate than those in the low-stress condition, while children's accuracy did not differ across stress conditions. Also, among adolescents, accuracy and errors were higher for the negative than positive words, while in children, word valence was unrelated to accuracy. Finally, increases in children's and adolescents' cortisol responses, especially in the high-stress condition, were related to greater accuracy but not false memories and only for positive emotional words. Findings suggest that stress at encoding, as well as the emotional content of to-be-remembered information, may influence memory in different ways across development, highlighting the need for greater complexity in existing models of true and false memory formation.

Optimal husbandry of hatchling Eastern Indigo Snakes (Drymarchon couperi) during a captive head-start program.

Optimal husbandry techniques are desirable for any headstart program, but frequently are unknown for rare species. Here we describe key reproductive variables and determine optimal incubation temperature and diet diversity for Eastern Indigo Snakes (Drymarchon couperi) grown in laboratory settings. Optimal incubation temperature was estimated from two variables dependent on temperature, shell dimpling, a surrogate for death from fungal infection, and deviation of an egg from an ovoid shape, a surrogate for death from developmental anomalies. Based on these relationships and size at hatching we determined optimal incubation temperature to be 26°C. Additionally, we used incubation data to assess the effect of temperature on duration of incubation and size of hatchlings. We also examined hatchling diets necessary to achieve optimal growth over a 21-month period. These snakes exhibited a positive linear relationship between total mass eaten and growth rate, when individuals were fed less than 1711 g of prey, and displayed constant growth for individuals exceeding 1711 g of prey. Similarly, growth rate increased linearly with increasing diet diversity up to a moderately diverse diet, followed by constant growth for higher levels of diet diversity. Of the two components of diet diversity, diet evenness played a stronger role than diet richness in explaining variance in hatchling growth. These patterns document that our goal of satiating snakes was achieved for some individuals but not others and that diets in which total grams consumed over the first 21 months of life is distributed equivalently among at least three prey genera yielded the fastest growth rates for hatchling snakes.

Experimental manipulation of the Trier Social Stress Test-Modified (TSST-M) to vary arousal across development.

Reliable laboratory protocols manipulating the intensity of biobehavioral arousal for children are uncommon, and those available have minimal converging evidence of their efficacy in manipulating arousal across multiple biological systems. This report presents two studies of the efficacy of the modified Trier Social Stress Test (TSST-M). In Study 1, sixty-three 7-15-year olds, and 19 young adults (18-25 yrs) completed the TSST-M. Comparable reactivity across age groups was observed for salivary cortisol, salivary alpha-amylase (sAA), pre-ejection period (PEP) and respiratory sinus arrhythmia (RSA), whereas self-reported stress was higher in adults compared to children. In Study 2, eighty-four 7-8-year olds and eighty-six 12-15-year olds were randomly assigned to a standard or low-stress TSST-M condition. Cortisol and self-reported stress responses were higher in the standard compared to the low-stress condition. In contrast, sAA and PEP were higher in the low-stress condition and RSA responses were comparable between the two TSST-M conditions. In addition, age group differences emerged in Study 2, though never in conjunction with the TSST-M manipulation. To test, refine, and advance theory about the implications of stress for child development, laboratory tasks that manipulate and enable assessment of biobehavioral arousal in children are needed.

Disclosure Suspicion Bias and Abuse Disclosure: Comparisons Between Sexual and Physical Abuse.

Prior research has found that children disclosing physical abuse appear more reticent and less consistent than children disclosing sexual abuse. Although this has been attributed to differences in reluctance, it may also be due to differences in the process by which abuse is suspected and investigated. Disclosure may play a larger role in arousing suspicions of sexual abuse, while other evidence may play a larger role in arousing suspicions of physical abuse. As a result, children who disclose physical abuse in formal investigations may be doing so for the first time, and they may be more reluctant to provide details of the abuse. We examined abuse disclosure and evidence in comparable samples of court-substantiated physical (n = 33) and sexual (n = 28) abuse. Consistent with predictions, the likelihood that the child had disclosed abuse before an investigation began was lower in physical (27%) than that in sexual (67%) abuse cases, and there was more nondisclosure evidence of abuse in physical abuse cases. These findings have implications for understanding the dynamics and meaning of disclosure in cases involving different types of abuse.

NPM-RAR, not the RAR-NPM reciprocal t(5;17)(q35;q21) acute promyelocytic leukemia fusion protein, inhibits myeloid differentiation.

The t(5;17) variant of acute promyelocytic leukemia (APL) fuses the nucleophosmin (NPM) gene at 5q35 with the retinoic acid receptor alpha (RARA) at 17q12-22. We have previously shown that leukemic cells express both NPM-RAR and RAR- NPM reciprocal translocation products. In this study we investigated the potential role of both proteins in modulating myeloid differentiation. Expression of NPM-RAR inhibited vitamin D3/transforming growth factor ß (TGFß)-mediated differentiation of U937 cells by more than 50%. In contrast, RAR-NPM expression did not alter vitamin D3/TGFß-induced differentiation of U937 clones. These results indicate that NPM-RAR, not RAR-NPM, is the prime mediator of myeloid differentiation arrest in t(5;17) APL.

Effects of stress on memory in children and adolescents: testing causal connections.

Although a sizeable body of research has examined children's memory for stressful prior experiences, relatively few studies have experimentally manipulated stress during a to-be-remembered event to draw causal inferences about the effects of stress, especially across wide age ranges. We exposed children and adolescents to a more or a less arousing version of the Trier Social Stress Test-Modified (TSST-M), a widely used laboratory stress task. Two weeks later, we tested their memory for what happened. Interviewers behaved in a supportive or non-supportive manner. In adolescents, those who completed the high-arousal TSST-M provided fewer correct responses to recognition questions and fewer incorrect responses to misleading questions for which any answer would have been incorrect, compared to those who completed the lower-arousal TSST-M. Thus, arousal seemed to have reduced the adolescents' willingness to answer questions rather than having influenced their memory per se. In children, across TSST-M conditions, greater physiological arousal during the TSST-M predicted enhanced recall. Finally, interviewer support reduced the amount of factual information provided in free recall but increased correct responses to misleading questions. Results highlight the complex ways in which event stress and interviewer demeanour shape recounting of prior experiences across development.

Physical activity and depressive symptoms in older adults.

OBJECTIVES: Depressive symptoms are prevalent in older adults, and physical activity (PA) may have beneficial effects on depression. The purpose of this study was to explore the association between physical activity and depressive symptoms, taking into account demographic factors, and the associations between selected demographic factors and physical activity levels in community-dwelling older adults (age ≥ 60 years). METHODS: Data were drawn from the National Health and Nutrition Examination Survey 2005-2006. Descriptive statistics and logistic models were used in data analysis. RESULTS: Four percent of participants reported moderate depressive symptoms, and 24% of subjects exhibited sedentary PA. Factors associated with increased risk of moderate depression included age, sedentary PA, and chronic medical conditions (ps < 0.05). Sedentary PA was significantly associated with age, race, education, BMI, smoking status, alcohol use, and taking psychotropics (ps < 0.05). DISCUSSION: PA is a protective factor for depression in older adults, and clinical implications to encourage PA are discussed.

Stress, interviewer support, and children's eyewitness identification accuracy.

Few studies have investigated how stress affects eyewitness identification capabilities across development, and no studies have investigated whether retrieval context in conjunction with stress affects accuracy. In this study, one hundred fifty-nine 7- to 8- and 12- to 14-year-olds completed a high- or low-stress laboratory protocol during which they interacted with a confederate. Two weeks later, they attempted to identify the confederate in a photographic lineup. The lineup administrator behaved in either a supportive or a nonsupportive manner. Participants who experienced the high-stress event and were questioned by a supportive interviewer were most accurate in rejecting target-absent lineups. Results have implications for debates about effects of stress on eyewitness recall, how best to elicit accurate identifications in children, and developmental changes in episodic mnemonic processes.

Interaction with RXR is necessary for NPM-RAR-induced myeloid differentiation blockade.

The t(5;17)(q35;q21) APL variant results in expression of a fusion protein linking the N-terminus of nucleophosmin (NPM) to the C-terminus of the retinoic acid receptor alpha (RAR). We have previously shown that NPM-RAR is capable of binding to DNA either as a homodimer or heterodimer with RXR. To determine the biological significance of NPM-RAR/RXR interaction, we developed two mutants of NPM-RAR that showed markedly diminished ability to bind RXR. U937 subclones expressing the NPM-RAR mutants showed significantly less inhibition of vitamin D3/TGFbeta-induced differentiation, compared with NPM-RAR. These results support the hypothesis that RXR interaction is necessary for NPM-RAR-mediated myeloid maturation arrest.

Stress at encoding, context at retrieval, and children's narrative content.

Research concerning the relations between stress and children's memory has been primarily correlational and focused on memory volume and accuracy. In the current study, we experimentally manipulated 7- and 8-year-olds' and 12- to 14-year-olds' experienced stress during a to-be-remembered event to examine the effects of stress on the content of their memory. We further manipulated the degree of interviewer support at retrieval to determine whether it moderated the effects of stress at encoding on memory. Children's age, gender, stress at encoding, and interviewer support all influenced the type of information included in their narrative reports. Most notably, across ages, children who experienced a more stressful event but were questioned in a supportive manner provided the largest ratio of terms representing internal states such as those about cognitions and emotions. Results suggest that how children process past events may be influenced by both the nature of the event itself and the context within which it is recalled.

Negative emotions and risk for type 2 diabetes among Korean immigrants.

PURPOSE: The purpose of this study is to examine the relationship between negative emotions and bio-behavioral risk factors among Korean immigrants at risk for type 2 diabetes (T2DM). METHODS: Data were collected from 148 Korean immigrant adults who are "at risk" for T2DM as defined by having family history of T2DM in first-degree relatives, body mass index greater than 23, or history of gestational diabetes in women. Participants completed questionnaires and underwent biological measures. Negative emotions included feeling nervous, hopeless, restless, anxious, and stressed as well as depressive symptoms. RESULTS: High percentages of participants had T2DM risk factors including overweight, greater than normal waist to hip ratio, and blood glucose readings that are indicative of T2DM. Feeling stressed was the most commonly reported negative emotion (66%), followed by feeling anxious (51%), restless (38%), nervous (30%), and hopeless (13%). Experience of negative emotions was significantly related to behavioral risk factors; higher levels of experiencing negative emotions were related to increased soda intake and a decreased likelihood of doing at least 10 minutes of moderate exercise. Stress and anxiety were each negatively related to moderate exercise, and depressive symptoms were negatively related to both moderate and vigorous exercise. No significant relationship was found between negative emotions and biological risk factors. CONCLUSIONS: Findings suggest that negative emotions, individually and taken together, may be related to T2DM risk behaviors in high-risk Korean immigrants. Behavioral interventions to prevent T2DM in this population should consider assessing and addressing negative emotions.