An In Vivo Model of Human Macrophages in Metastatic Melanoma.

Despite recent therapeutic progress, advanced melanoma remains lethal for many patients. The composition of the immune tumor microenvironment (TME) has decisive impacts on therapy response and disease outcome, and high-dimensional analyses of patient samples reveal the heterogeneity of the immune TME. Macrophages infiltrate TMEs and generally associate with tumor progression, but the underlying mechanisms are incompletely understood. Because experimental systems are needed to elucidate the functional properties of these cells, we developed a humanized mouse model reconstituted with human immune cells and human melanoma. We used two strains of recipient mice, supporting or not supporting the development of human myeloid cells. We found that human myeloid cells favored metastatic spread of the primary tumor, thereby recapitulating the cancer-supportive role of macrophages. We next analyzed the transcriptome of human immune cells infiltrating tumors versus other tissues. This analysis identified a cluster of myeloid cells present in the TME, but not in other tissues, which do not correspond to canonical M2 cells. The transcriptome of these cells is characterized by high expression of glycolytic enzymes and multiple chemokines and by low expression of gene sets associated with inflammation and adaptive immunity. Compared with humanized mouse results, we found transcriptionally similar myeloid cells in patient-derived samples of melanoma and other cancer types. The humanized mouse model described here thus complements patient sample analyses, enabling further elucidation of fundamental principles in melanoma biology beyond M1/M2 macrophage polarization. The model can also support the development and evaluation of candidate antitumor therapies.

Astrocytic laminin-211 drives disseminated breast tumor cell dormancy in brain.

Although dormancy is thought to play a key role in the metastasis of breast tumor cells to the brain, our knowledge of the molecular mechanisms regulating disseminated tumor cell (DTC) dormancy in this organ is limited. Here using serial intravital imaging of dormant and metastatic triple-negative breast cancer lines, we identify escape from the single-cell or micrometastatic state as the rate-limiting step towards brain metastasis. We show that every DTC occupies a vascular niche, with quiescent DTCs residing on astrocyte endfeet. At these sites, astrocyte-deposited laminin-211 drives DTC quiescence by inducing the dystroglycan receptor to associate with yes-associated protein, thereby sequestering it from the nucleus and preventing its prometastatic functions. These findings identify a brain-specific mechanism of DTC dormancy and highlight the need for a more thorough understanding of tumor dormancy to develop therapeutic approaches that prevent brain metastasis.

Detection of engineered T cells in FFPE tissue by multiplex in situ hybridization and immunohistochemistry.

Identifying engineered T cells in situ is important to understand the location, persistence, and phenotype of these cells in patients after adoptive T cell therapy. While engineered cells are routinely characterized in fresh tissue or blood from patients by flow cytometry, it is difficult to distinguish them from endogenous cells in formalin-fixed, paraffin-embedded (FFPE) tissue biopsies. To overcome this limitation, we have developed a method for characterizing engineered T cells in fixed tissue using in situ hybridization (ISH) to the woodchuck hepatitis post-transcriptional regulatory element (WPRE) common in many lentiviral vectors used to transduce chimeric antigen receptor T (CAR-T) and T cell receptor T (TCR-T) cells, coupled with alternative permeabilization conditions that allows subsequent multiplex immunohistochemical (mIHC) staining within the same image. This new method provides the ability to mark the cells by ISH, and simultaneously stain for cell-associated proteins to immunophenotype CAR/TCR modified T cells within tumors, as well as assess potential roles of these cells in on-target/off-tumor toxicity in other tissue.

Blood pressure control in continuous flow left ventricular assist devices: efficacy and impact on adverse events.

BACKGROUND: Continuous flow (CF) left ventricular assist devices (LVAD) are afterload sensitive and therefore pump performance is affected by hypertension. In addition, poorly controlled hypertension may increase the risk of aortic insufficiency (AI) and stroke. Blood pressure regimens after CF LVAD have not been studied and their impact on rates of AI and stroke are unknown. METHODS: Patients who had CF LVAD at a single center and were supported greater than 30 days were included. Blood pressure was monitored at home by Doppler. Outpatient management of blood pressure was conducted according to a predefined institutional protocol (target mean arterial pressure ≤ 80 mm Hg). RESULTS: A total of 96 patients were included. At the end of follow-up, 25 patients were not on an antihypertensive drug, of these 9 died. Of the 74% receiving antihypertensives, 54% required 1 medication, 34% were on 2, 10% were on 3, and 3% were on 4 or more. Angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers (85% of patients on an antihypertensive) and beta blockers (30%) were the most commonly prescribed medications. There was a significantly higher neurologic event rate in those on no antihypertensives compared with those on antihypertensives (p = 0.009). Only 3% of patients with no or mild AI at baseline progressed to develop moderate or greater AI after a mean of 201 days of follow-up. CONCLUSIONS: Blood pressure control can be achieved in patients with CF LVADs, with the majority of patients requiring only 1 or 2 antihypertensives.

Mental- and physical-health indicators and sexually explicit media use behavior by adults.

INTRODUCTION: Converging evidence from culturally diverse contexts indicates that sexually explicit media use behavior (SEMB; i.e., pornography consumption) is associated with risky sexual health perceptions and behaviors, many that involve high risks of HIV/STD transmission. AIM: Essentially unexplored, and the focus here, are potential relationships between SEMB and nonsexual mental- and physical-health indicators. MAIN OUTCOME MEASURE: Variability in six continuously measured health indicators (depressive symptoms, mental- and physical-health diminished days, health status, quality of life, and body mass index) was examined across two levels (users, nonusers) of SEMB. METHODS: A sample of 559 Seattle-Tacoma Internet-using adults was surveyed in 2006. Multivariate general linear models parameterized in a SEMB by respondent gender (2 × 2) factorial design were computed incorporating adjustments for several demographics. RESULTS: SEMB was reported by 36.7% (n = 205) of the sample. Most SEMB users (78%) were men. After adjusting for demographics, SEMB users, compared to nonusers, reported greater depressive symptoms, poorer quality of life, more mental- and physical-health diminished days, and lower health status. CONCLUSIONS: The findings show that mental- and physical-health indicators vary significantly across SEMB, suggesting the value of incorporating these factors in future research and programmatic endeavors. In particular, the findings suggest that evidence-based sexual health promotion strategies simultaneously addressing individuals' SEMB and their mental health needs might be a useful approach to improve mental health and address preventable sexual health outcomes associated with SEMB.

Health information-seeking behaviors, health indicators, and health risks.

OBJECTIVES: We examined how different types of health information-seeking behaviors (HISBs)-no use, illness information only, wellness information only, and illness and wellness information combined-are associated with health risk factors and health indicators to determine possible motives for health information seeking. METHODS: A sample of 559 Seattle-Tacoma area adults completed an Internet-based survey in summer 2006. The survey assessed types of HISB, physical and mental health indicators, health risks, and several covariates. Covariate-adjusted linear and logistic regression models were computed. RESULTS: Almost half (49.4%) of the sample reported HISBs. Most HISBs (40.6%) involved seeking a combination of illness and wellness information, but both illness-only (28.6%) and wellness-only (30.8%) HISBs were also widespread. Wellness-only information seekers reported the most positive health assessments and the lowest occurrence of health risk factors. An opposite pattern emerged for illness-only information seekers. CONCLUSIONS: Our findings reveal a unique pattern of linkages between the type of health information sought (wellness, illness, and so on) and health self-assessment among adult Internet users in western Washington State. These associations suggest that distinct health motives may underlie HISB, a phenomenon frequently overlooked in previous research.

Streptococcosis in farmed Litopenaeus vannamei: a new emerging bacterial disease of penaeid shrimp.

Presumptive systemic streptococcal infections were detected histologically in farmed Litopenaeus vannamei juveniles submitted from a Latin American country and the bacteria isolated. Characterization work demonstrated that the Gram-positive cocci form chains, grow aerobically and anaerobically, are oxidase- and catalase-negative, non-hemolytic, non-motile, Lancefield Group B positive and PCR positive when amplified with a universal streptococcal primer set. Differing Streptococcus identifications were obtained using API 20 Strep and Biolog systems, the former identifying the isolate as S. uberis and the latter as S. parauberis. Injection of specific pathogen-free (SPF) L. vannamei with the bacteria resulted in 100% mortality by 3 d post-injection with successful recovery of the agent from moribund test shrimp hemolymph samples. The recovered isolate was used in per os and waterborne exposure studies of SPF L. vannamei with mortalities ranging from 40 to 100% and 80 to 100%, respectively. Histologic analysis of 5 to 8 moribund shrimp from each exposure method demonstrated that all contained a severe bacteremia characterized by numerous free cocci within the hemolymph and aggregates of vacuolated hemocytes with notable intravacuolar cocci. This unique lesion type was most pronounced within the lymphoid organ and considered pathodiagnostic for this disease. Experimentally induced lesions were identical to those in naturally infected farmed shrimp and the Streptococcus sp. responsible was re-isolated, fulfilling Koch's postulates. Five freeze/thaw cycles of 10 experimentally infected shrimp were performed over a 2 mo period and the bacteria successfully cultured from all shrimp at each interval. These collective findings describe the first reported case of streptococcosis in marine penaeid shrimp in the Western Hemisphere and indicate that the agent may be disseminated via live or frozen infected shrimp.

Health-risk correlates of video-game playing among adults.

BACKGROUND: Although considerable research suggests that health-risk factors vary as a function of video-game playing among young people, direct evidence of such linkages among adults is lacking. PURPOSE: The goal of this study was to distinguish adult video-game players from nonplayers on the basis of personal and environmental factors. It was hypothesized that adults who play video games, compared to nonplayers, would evidence poorer perceptions of their health, greater reliance on Internet-facilitated social support, more extensive media use, and higher BMI. It was further hypothesized that different patterns of linkages between video-game playing and health-risk factors would emerge by gender. METHODS: A cross-sectional, Internet-based survey was conducted in 2006 with a sample of adults from the Seattle-Tacoma area (n=562), examining health risks; media use behaviors and perceptions, including those related to video-game playing; and demographics. Statistical analyses conducted in 2008 to compare video-game players and nonplayers included bivariate descriptive statistics, stepwise discriminant analysis, and ANOVA. RESULTS: A total of 45.1% of respondents reported playing video games. Female video-game players reported greater depression (M=1.57) and poorer health status (M=3.90) than female nonplayers (depression, M=1.13; health status, M=3.57). Male video-game players reported higher BMI (M=5.31) and more Internet use time (M=2.55) than male nonplayers (BMI, M=5.19; Internet use, M=2.36). The only determinant common to female and male video-game players was greater reliance on the Internet for social support. CONCLUSIONS: A number of determinants distinguished video-game players from nonplayers, and these factors differed substantially between men and women. The data illustrate the need for further research among adults to clarify how to use digital opportunities more effectively to promote health and prevent disease.

Developing healthy kids in healthy communities: eight evidence-based strategies for preventing high-risk behaviour.

Australian youth engage in behaviour that threatens their health and wellbeing. National surveys report that about a third of young Australians have tried an illicit drug. High rates of substance use and risky sexual behaviour among young Australians suggest that effective prevention efforts based on empirical evidence need to be expanded. Church-associated organisations are an untapped resource that could be used to improve the health and welfare of young people. We describe eight evidence-based elements to consider in designing strategies to prevent high-risk behaviour in young people.

Effect of transducer velocity on intramuscular temperature during a 1-MHz ultrasound treatment.

STUDY DESIGN: A 3 x 2 repeated-measures design was used. The independent variables were transducer velocity (2-3 cm/s, 4-5 cm/s, and 7-8 cm/s) and time (pretreatment and posttreatment). OBJECTIVE: To determine if transducer velocity of a 1-MHz ultrasound treatment affects intramuscular tissue temperature. BACKGROUND: Most authors advocate ultrasound transducer velocities of 2 to 4 cm/s within an area of 2 to 3 times the effective radiating area or 2 times the size of the transducer head. However, a much faster rate of application (approximately 7-8 cm/s) is often observed in clinical settings. METHODS AND MEASURES: Eleven healthy screened volunteers (9 males, 2 females; mean +/- SD age, 22.6 +/- 1.7 years; mean +/- SD height, 175.7 +/- 13.7 cm; mean +/- SD body mass, 82.5 +/- 19.5 kg) were randomly assigned to a treatment order with all conditions administered during a single testing session. Each transducer velocity condition was administered for 10 minutes, using 1-MHz ultrasound with a 100% continuous duty cycle at an intensity of 1.5 W/cm2 over an area twice the size of the transducer head. After the first treatment, the 2 remaining subsequent velocity conditions were administered after the intramuscular temperature returned to within +/- 0.3 degrees C of the initial pretreatment temperature for 5 minutes. The dependent variable was left triceps surae muscle temperature measured at 3 cm below one half the measured skinfold thickness. RESULTS: Temperature increase across the 3 velocities was within 0.4 degrees C (F2.20 = 0.07, P = .93). Posttreatment values (mean +/- SD) ranged from 42.7 degrees C +/- 2.3 degrees C for the slowest velocity to 43.1 degrees C +/- 1.4 degrees C for the fastest velocity. Temperature increase was significant for time (F1.01 = 155.68, P<.00001), increasing from 37.8 degrees C +/- 0.8 degrees C pretreatment to 42.9 degrees C +/- 1.9 degrees C after treatment. CONCLUSION: Very similar intramuscular temperature increases can be observed among ultrasound treatments (10-minute duration, 1-MHz frequency, 100% continuous duty cycle, 1.5 W/cm2 intensity, within an area twice the size of the transducer head), with transducer velocities of 2 to 3, 4 to 5, and 7 to 8 cm/s.