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.

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.

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.

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.

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 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.

Multicentric neurofibromatosis with rectal prolapse in a California sea lion (Zalophus californianus).

An approximately 31-yr-old California sea lion (Zalophus californianus) with a history of chronic visual impairment and corneal disease presented with slow onset, progressive neurologic deficits. Treatment for rear flipper paresis was not effective and the animal was euthanatized. Histopathologic findings included hepatocellular and biliary neoplasia, ocular amyloidosis, adrenal adenoma and pheochromocytoma, and spinal cord changes consistent with multicentric neurofibromatosis. This is the first documentation of these conditions in a California sea lion.

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.

Ethnic and language disparities in diabetes care among California residents.

OBJECTIVE: We examined ethnic and language disparities in diabetes care and management among California residents with type 2 diabetes based on ethnicity and English-language proficiency. METHODS: Data were drawn from the 2007 California Health Interview Survey with a total of 3,531 Asian, Latino, and Caucasian adults with diabetes. Latino and Asian groups were subdivided by their limited English proficiency (LEP) level. Population-weighted regression analyses were conducted to examine group differences, controlling for socioeconomic and clinical variables. RESULTS: Latino English Proficient (EP) and Latino LEP respondents received fewer hemoglobin A1c checks (EP: b = -0.11, P < .05; LEP: b = -0.27, P < .01) than Caucasians. Latino and Asian LEP respondents checked their glucose less frequently than Caucasians (Latino LEP: b = -0.49, P < .05; Asian LEP: b = -0.79, P < .01). Asian LEP respondents were less likely to receive feet checks than Caucasians (Asian LEP: b = -.52, P < .001). Asian LEP respondents received significantly fewer feet checks than Asian EP respondents (P < .05). CONCLUSIONS: Ethnic disparities in disease management exist among California residents. However, beyond ethnicity, English proficiency should be taken into account when examining diabetes management among minority groups. Diabetic Californians who belong to ethnic minorities and speak limited English, particularly Asians, are less likely to receive the standard of care for diabetes than English proficient Caucasians. From a policy perspective, care should be taken to ensure that adequate information about diabetes management is available in multiple languages for patients with limited English skills.

The role of cortisol reactivity in children's and adults' memory of a prior stressful experience.

The purpose of this study was to identify whether cortisol reactivity to a stressful laboratory event was related to children's memory of that event and to determine whether this relation was comparable to that observed in adults. Nine- to 12-year-olds and young adults completed an impromptu speech and math task during which repeated cortisol samples and self-reported stress ratings were collected. Two weeks later, participants' memory for the tasks was examined. Greater cortisol reactivity was associated with enhanced memory, most prominently in children. Self-reported stress was unrelated to memory. Findings reveal that an important mechanism underlying the association between emotion and memory in adults, namely activation of the hypothalamic pituitary adrenal axis, appears to operate similarly in late childhood. Findings also demonstrate that positive associations between cortisol reactivity and memory are evident when the event that actually elicited that reactivity serves as the to-be-remembered event.

Clinical management of a western lowland gorilla (Gorilla gorilla gorilla) with a cardiac resynchronization therapy device.

A 24-yr-old, male western lowland gorilla (Gorilla gorilla gorilla) was diagnosed with congestive heart failure using transesophageal and transthoracic echocardiology. New York Heart Association (NYHA) Class III was assigned to the severity of the condition. Over 16 mo, this progressed to NYHA Class IV despite increasing medical therapy. Repeated evaluations suggested that implantation of a cardiac resynchronization therapy device with a defibrillator (CRT-D) could benefit this animal based on clinical signs and underlying evidence of dyssynchrony and suspected fibrotic myocardial disease. Surgical implantation of leads into the right atrium, right ventricle, and left ventricle was accomplished. The CRT-D device was placed under the thoracic pectoral muscles during an initial surgical procedure. Improvement in the gorilla's clinical condition after implantation of the CRT-D device was immediate and dramatic. Subsequent scanning of the device was accomplished through operant conditioning. The data from these device interrogations included stored and real-time cardiac data, which were used to minimize recognized environmental stressors and change device settings. Over 4 yr, case management was critical to successful device use in treatment of the clinical disease. This involved medications, training for device interrogation, exercise to increase activity and improve body condition, and phlebotomy attempts. Dietary management was necessary to manipulate caloric and sodium intake and encourage medication compliance. Cardiac resynchronization therapy device implantation, although requiring specialized equipment and surgical skill, appears to be a viable option for treatment of fibrosing cardiomyopathy with systolic dysfunction in gorillas refractory to medical management. In addition to treatment, this device provides cardiovascular data at rest that could allow for early diagnosis and treatment of gorillas with this and other cardiac conditions in the future. This describes the comprehensive medical, husbandry, and training techniques necessary to successfully manage this intense clinical case in conjunction with intracardiac device therapy.

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.

Surgical implantation of a cardiac resynchronization therapy device in a western lowland gorilla (Gorilla gorilla gorilla) with fibrosing cardiomyopathy.

A 24-yr-old, male western lowland gorilla (Gorilla gorilla gorilla) was diagnosed in March of 2003 with congestive heart failure (CHF). Transesophageal and transthoracic echocardiography demonstrated global left and right ventricular hypokinesia with a left ventricular ejection fraction of 0.20. At the time of diagnosis, the animal exhibited symptoms and signs of CHF with minimal exertion (New York Heart Association class III). Over a 16-mo period, the severity of CHF progressed to class IV (resting signs and symptoms) despite angiotensin-converting enzyme inhibition, beta-blockers, and diuretics. Because of intractable CHF and a QRS duration that was markedly prolonged compared with the normal range for this species, a cardiac resynchronization therapy (CRT) device was implanted using implantation techniques based on human surgical procedures. Placement of the right ventricular, right atrial, and left ventricular leads and pulse generator were accomplished in 5.5 hr. Telemetry of the device postoperatively via wand or remote radio frequency has allowed for noninvasive programming and interrogation. The clinical improvement in CHF with this therapy was immediate and dramatic for this animal. Six months after CRT device implantation, the device leads became dislodged during an altercation with another gorilla, with the rapid development of CHF upon cessation of biventricular pacing. A second procedure to replace the leads returned the gorilla to his previous level of activity. In 2007, the pulse generator was electively replaced for battery depletion with a device capable of remote radiofrequency programming and interrogation. CRT implantation, although requiring specialized equipment and surgical skill, appears to be a viable option for treatment of dilated cardiomyopathy in gorillas.

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 NPM-RAR fusion protein associated with the t(5;17) variant of APL does not interact with PML.

The PML protein localizes to regions of the nucleus known as nuclear bodies or PODs. However, in t(15;17) Acute Promyelocytic Leukemia (APL) blasts, PML is found in a micro-punctate pattern. In order to test the hypothesis that delocalization of PML from PODs is necessary for APL, we investigated the interaction of the t(5;17) APL fusion protein NPM-RAR with PML. NPM-RAR localizes diffusely throughout the nucleoplasm. NPM-RAR does not alter the localization of PML in transfected HeLa cells, and does not associate with PML in vitro. These studies suggest that NPM-RAR does not interact with PML.

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.