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Introduction/purpose Most US adults (54%) do not meet the minimum exercise recommendations by the American College of Sports Medicine. Neuromuscular electrical stimulation (NMES) is a novel alternate strategy to induce muscle contraction. However, the effectiveness of NMES to improve insulin sensitivity and energy expenditure is unclear. The purpose of this study was to investigate the effects of 4 weeks of NMES on glucose tolerance in a sedentary overweight or obese population. Methods Participants (n = 10; age: 36.8 ± 3.8 years; BMI = 32 ± 1.3 kg/m2) were randomized into either control or NMES group. All participants received bilateral quadriceps stimulation (12 sessions; 30 min/session; three times/week at 50 Hz and 300 µs pulse width) altering pulse amplitude to either provide low-intensity sensory level (control; tingling sensation) or at high-intensity neuromuscular level (NMES; maximum tolerable levels with visible muscle contraction). Glucose tolerance was assessed by a 3-h oral glucose tolerance test (OGTT), and substrate utilization was measured by indirect calorimetry and body composition via dual X-ray absorptiometry at baseline and after 4 weeks of NMES intervention. Results Control and NMES groups had comparable fasting blood glucose, glucose tolerance, substrate utilization, and muscle mass at baseline. Four weeks of NMES resulted in a significant improvement in glucose tolerance measured by OGTT, whereas no change was observed in the control group. There was no change in substrate utilization and muscle mass in both control and NMES groups. Conclusion NMES is a novel and effective strategy to improve glucose tolerance in an at-risk overweight or obese sedentary population.

BACKGROUND Hypertension is a major contributor of various adverse health outcomes. Although prior studies have shown benefits of home blood pressure monitoring over office-based measurements, there is limited evidence comparing the effectiveness of whether a blood pressure monitor integrated into the electronic health record is superior to a non-integrated blood pressure monitor. OBJECTIVE In this paper, we describe the protocol for a pragmatic multisite implementation of a quality improvement initiative directly comparing integrated to non-integrated blood pressure monitors for hypertension improvement. METHODS This is a 12-month randomized, comparative effectiveness study conducted at three large academic health centers across California. The three sites will enroll a total of 660 participants (approximately n=220 per site) with 330 in the integrated blood pressure monitor arm and 330 in the non-integrated blood pressure control arm. The primary outcome of this study will be absolute difference in systolic blood pressure in mmHg from enrollment to 6 months. Secondary outcome measures include binary measures of hypertension (controlled vs uncontrolled), hypertension-related health complications, hospitalizations, and death. The list of possible participants will be generated from a central data warehouse. Randomization will occur after enrollment into the study. Participants will use their assigned blood pressure monitor and join site-specific hypertension interventions. Cross-site learning will occur at regular all-site meetings facilitated by the UCLA Value-Based Care Research Consortium. A pre-study and post-study questionnaire will be conducted to further evaluate participant perspectives regarding their blood pressure monitor. Linear mixed effects models will be used to compare the primary outcome measure between study arms. Mixed effects logistic regression models will be used to compare secondary outcome measures between study arms. RESULTS The study will start enrolling participants in the second quarter of 2023 and complete by first half 2024. Results will be published by end of 2024. CONCLUSIONS This pragmatic trial will contribute to the growing field of chronic care management using remote monitoring by answering whether a hypertension intervention coupled with an electronic health record integrated home blood pressure monitor improves patients’ hypertension better than a hypertension intervention with a non-integrated blood pressure monitor. The outcomes of this study may help health system decision-makers determine whether to invest in integrated blood pressure monitors for vulnerable patient populations. CLINICALTRIAL The ClinicalTrials.gov ID for this study is "NCT05390502." It was registered 05/20/2022. Protocol ID is 22000036.

Galpha and MEK1/2 inhibition in combination with Baf A1 and CQ in MP41 cells

Supplementary Materials and Methods

Purpose:Mutational activation of GNAQ or GNA11 (GNAQ/11), detected in >90% of uveal melanomas, leads to constitutive activation of oncogenic pathways, including MAPK and YAP. To date, chemo- or pathway-targeted therapies, either alone or in combination, have proven ineffective in the treatment of patients with metastatic uveal melanoma.Experimental Design:We tested the efficacy of chloroquine or hydroxychloroquine, in combination with MAPK pathway inhibition in GNAQ/11-mutated cells in vitro and in vivo and identified mechanisms of MEK1/2 inhibitor plus chloroquine-induced cytotoxicity.Results:Inhibition of GNAQ/11-mediated activation of MAPK signaling resulted in the induction of autophagy. Combined inhibition of Gα and autophagy or lysosome function resulted in enhanced cell death. Moreover, the combination of MEK1/2 inhibition, using trametinib, with the lysosome inhibitor, chloroquine, also increased cytotoxicity. Treatment of mice bearing GNAQ/11-driven melanomas with trametinib plus hydroxychloroquine resulted in inhibition of tumor growth and significantly prolonged survival. Interestingly, lysosomal- and autophagy-specific inhibition with bafilomycin A1 was not sufficient to promote cytotoxicity in combination with trametinib. However, the addition of YAP inhibition with trametinib plus bafilomycin A1 resulted in cell death at comparable levels to trametinib plus chloroquine (T/CQ) treatment. Furthermore, T/CQ-treated cells displayed decreased YAP nuclear localization and decreased YAP transcriptional activity. Expression of a constitutively active YAP5SA mutant conferred resistance to T/CQ-induced cell death.Conclusions:These results suggest that YAP, MEK1/2, and lysosome function are necessary and critical targets for the therapy of GNAQ/11-driven melanoma, and identify trametinib plus hydroxychloroquine as a potential treatment strategy for metastatic uveal melanoma.

Purpose:Mutational activation of GNAQ or GNA11 (GNAQ/11), detected in >90% of uveal melanomas, leads to constitutive activation of oncogenic pathways, including MAPK and YAP. To date, chemo- or pathway-targeted therapies, either alone or in combination, have proven ineffective in the treatment of patients with metastatic uveal melanoma.Experimental Design:We tested the efficacy of chloroquine or hydroxychloroquine, in combination with MAPK pathway inhibition in GNAQ/11-mutated cells in vitro and in vivo and identified mechanisms of MEK1/2 inhibitor plus chloroquine-induced cytotoxicity.Results:Inhibition of GNAQ/11-mediated activation of MAPK signaling resulted in the induction of autophagy. Combined inhibition of Gα and autophagy or lysosome function resulted in enhanced cell death. Moreover, the combination of MEK1/2 inhibition, using trametinib, with the lysosome inhibitor, chloroquine, also increased cytotoxicity. Treatment of mice bearing GNAQ/11-driven melanomas with trametinib plus hydroxychloroquine resulted in inhibition of tumor growth and significantly prolonged survival. Interestingly, lysosomal- and autophagy-specific inhibition with bafilomycin A1 was not sufficient to promote cytotoxicity in combination with trametinib. However, the addition of YAP inhibition with trametinib plus bafilomycin A1 resulted in cell death at comparable levels to trametinib plus chloroquine (T/CQ) treatment. Furthermore, T/CQ-treated cells displayed decreased YAP nuclear localization and decreased YAP transcriptional activity. Expression of a constitutively active YAP5SA mutant conferred resistance to T/CQ-induced cell death.Conclusions:These results suggest that YAP, MEK1/2, and lysosome function are necessary and critical targets for the therapy of GNAQ/11-driven melanoma, and identify trametinib plus hydroxychloroquine as a potential treatment strategy for metastatic uveal melanoma.

Multiple sclerosis (MS) is a neuro-inflammatory autoimmune disease of the central nervous system (CNS) that affects young adults. It is characterised by the development of demyelinating lesions and inflammation within the CNS. Although the causes of MS are still elusive, recent work using patient samples and experimental animal models has demonstrated a strong relationship between the gut microbiota and its contribution to CNS inflammation and MS. While there is no cure for MS, alteration of the gut microbiota composition through the use of probiotics is a very promising treatment. However, while most recent works have focused on the use of probiotics to modify pre-existing disease, little is known about its role in protecting from the establishment of MS. In this study, we determined whether colonisation with the probiotic bacterium Escherichia coli strain Nissle 1917 (EcN) could be used as a prophylactic strategy to prevent or alter the development of experimental autoimmune encephalomyelitis (EAE), a preclinical model of MS. We found that double gavage (two doses) of EcN before induction of EAE delayed disease onset and decreased disease severity. We also found that EcN-treated mice had decreased amounts of perivascular cuffing, CD4+ T cell infiltration into the CNS, together with significantly decreased absolute numbers of Th1 cells, and reduced activation of microglia. Although further studies are necessary to comprehend the exact protective mechanisms induced, our study supports a promising use of EcN as a probiotic for the prevention of MS.

Background Multiple sclerosis (MS) is an inflammatory demyelinating disease that affects 2.5 million people worldwide. Growing evidence suggests that perturbation of the gut microbiota, the dense collection of microorganisms that colonize the gastrointestinal tract, plays a functional role in MS. Indeed, specific gut-resident bacteria are altered in patients with MS compared to healthy individuals, and colonization of gnotobiotic mice with MS-associated microbiota exacerbates preclinical models of MS. However, defining the molecular mechanisms by which gut commensals can remotely affect the neuroinflammatory process remains a critical gap in the field. Methods We utilized monophasic experimental autoimmune encephalomyelitis (EAE) in C57BL/6J mice and relapse-remitting EAE in SJL/J mice to test the effects of the products from a human gut-derived commensal strain of Lactobacillus paracasei (Lb). Results We report that Lb can ameliorate preclinical murine models of MS with both prophylactic and therapeutic administrations. Lb ameliorates disease through a Toll-like receptor 2-dependent mechanism via its microbe-associated molecular patterns that can be detected in the systemic circulation, are sufficient to downregulate chemokine production, and can reduce immune cell infiltration into the central nervous system (CNS). In addition, alterations in the gut microbiota mediated by Lb-associated molecular patterns are sufficient to provide partial protection against neuroinflammatory diseases. Conclusions Local Lb modulation of the gut microbiota and the shedding of Lb-associated molecular patterns into the circulation may be important physiological signals to prevent aberrant peripheral immune cell infiltration into the CNS and have relevance to the development of new therapeutic strategies for MS.

Using in-depth, face-to-face personal interviews of 25 formerly incarcerated participants, this study examines former inmates’ perceptions of prison gang affiliation, factors that shaped their deci...

We present an overview and first results from a $M$-band spectroscopic survey of planet-forming disks performed with iSHELL on IRTF, using two slits that provide resolving power R $\approx$ 60,000-92,000 (5-3.3 km/s). iSHELL provides a nearly complete coverage at 4.52-5.24 $μ$m in one shot, covering $>50$ lines from the R and P branches of $^{12}$CO and $^{13}$CO for each of multiple vibrational levels, and providing unprecedented information on the excitation of multiple emission and absorption components. Some of the most notable new findings of this survey are: 1) the detection of two CO Keplerian rings at $, Accepted for publication on The Astronomical Journal

Viral encephalitis is a major cause of morbidity and mortality, but the manifestation of disease varies greatly between individuals even in response to the same virus. Microglia are professional antigen presenting cells that reside in the central nervous system (CNS) parenchyma that are poised to respond to viral insults. However, the role of microglia in initiating and coordinating the antiviral response is not completely understood. Utilizing Theiler’s murine encephalomyelitis virus (TMEV), a neurotropic picornavirus, and PLX5622, a small molecule inhibitor of colony-stimulating factor 1 receptor (CSF1R) signaling that can deplete microglia in the CNS; we investigated the role of the CSF1R-microglia axis in neurotropic picornavirus infection of C57BL/6J and SJL/J mice. These mouse strains differ in their ability to clear TMEV and exhibit different neurological disease in response to TMEV infection. CSF1R antagonism in C57BL/6J mice, which normally clear TMEV in the CNS, led to acute fatal encephalitis. In contrast, CSF1R antagonism in SJL/J mice, which normally develop a chronic CNS TMEV infection, did not result in acute encephalitis, but exacerbated TMEV-induced demyelination. Immunologically, inhibition of CSF1R in C57BL/6J mice reduced major histocompatibility complex II expression in microglia, decreased the proportion of regulatory T cells in the CNS, and upregulated proinflammatory pathways in CNS T cells. Acute CSF1R inhibition in SJL/J mice had no effect on microglial MHC-II expression and upregulated anti-inflammatory pathways in CNS T cells, however chronic CSF1R inhibition resulted in broad immunosuppression. Our results demonstrate strain-specific effects of the CSF1R-microglia axis in the context of neurotropic viral infection as well as inherent differences in microglial antigen presentation and subsequent T cell crosstalk that contribute to susceptibility to neurotropic picornavirus infection.

Exploratory and confirmatory factor analyses are commonly employed to identify and explore underlying factor structures. Unfortunately, when variable selection is involved, results often fluctuate across studies making it difficult to determine the “best” and most replicable factor structure. This study proposes a new factor analysis variable selection algorithm called the Replicable Factor Analytic Solutions (RFAS) that incorporates sound statistical and psychometric practices when selecting the final factor structure, while simultaneously examining the observed variables and factor structures replicability. This article outlines the algorithm development and rationale for each decision in the algorithm’s development. An example using simulated and empirical data is also provided to display the algorithm results and delineate the utility of these results for future analysts.

There is perhaps no disease in the modern era as geographically ubiquitous as cancer. Despite the high rates of the disease, particularly in developed countries, there is some geographic va...

This study examined three methods for retrospectively identifying infection in emergency department (ED) patients: modified objective definitions of infection (MODI) from the CDC/NHSN, physician adjudication determination of infection, and ED treating physician behavior.This study used a subset of data from a prospective sepsis trial. We used Fleiss's Kappa to compare agreement between two physicians retrospectively adjudicating infection based on the patient's medical record, modified infection definition from the CDC/NHSN, and ED treating physician behavior.Overall, there was similar agreement between physician adjudication of infection and MODI criteria (Kappa=0.59) compared to having two physicians independently identify infection through retrospective chart review (Kappa=0.58). ED treating physician behavior was a poorer proxy for infection when compared to the MODI criteria (0.41) and physician adjudication (Kappa = 0.50).Retrospective identification of infection poses a significant challenge in sepsis clinical trials. Using modified definitions of infection provides a standardized, less time consuming, and equally effective means of identifying infection compared to having multiple physicians adjudicate a patient's chart.

United Nations Police (UNPOL) officers who are deployed to United Nations Peacekeeping Missions worldwide must first undergo pre-deployment training. After arrival in the mission area, UNPOL officers go through another week of induction training. Both pre-deployment and induction training provide training and insight into the cultural paradigms of the population of the mission area. The idea being that the UNPOL officers must be prepared to work with host country nationals. The problem is that UNPOL missions are comprised of police officers from dozens of different nationalities. The hyper-diversity of an UNPOL organization means that UNPOL officer will encounter significant cross-cultural challenges simply working with each other. UNPOL officers never receive cultural training to facilitate their ability to work with people from all over the world. This chapter covers the newly identified and described concept of complex acculturation, which is the cultural phenomenon of having to adjust to multiple new cultures simultaneously. An understanding of complex acculturation could be a significant element in the improvement of UNPOL efficiency in peacekeeping missions around the world.

One of the primary functions of United Nations Police (UNPOL) officers in peacekeeping missions is the mentoring of local police forces in the mission area. UNPOL Mentors are tasked with motivating, training, developing, and promoting democratic police reform in the mission country’s organic police force. However, mentoring police officers from a country different than the UNPOL’s home country is more difficult and complex than domestic mentorship. This chapter explores the complexities of international police mentorship through lessons learned from the author’s experience as an UNPOL Mentor and Regional Commander in Haiti. The primary concepts and goals of UNPOL mentorship in a United Nations Peacekeeping mission are explored by using real-life situations to contextualize the primary goals of UNPOL mentorship.

The central nervous system (CNS) is an important regulator of the gastrointestinal tract, and CNS dysfunction can result in significant and disabling gastrointestinal symptom manifestation. For patients with neuroimmunologic and neuroinflammatory conditions, the recognition of gastrointestinal symptoms is under-appreciated, yet the gastrointestinal manifestations have a dramatic impact on quality of life. The current treatment strategies, often employed independently by the neurologist and gastroenterologist, raise the question of whether such patients are being treated optimally when siloed in one specialty. Neuroimmunogastroenterology lies at the borderlands of medical specialties, and there are few resources to guide neurologists in this area. Here, we provide an overview highlighting the potential mechanisms of crosstalk between immune-mediated neurological disorders and gastrointestinal dysfunction.

Most U.S. adults do not meet minimum exercise recommendations by ACSM. We and others have shown an increase in GLUT4 protein and glucose uptake with electrical stimulation induced muscle contraction using an in vitro cell culture model. However, effectiveness of neuromuscular electrical stimulation (NMES) in humans to improve insulin sensitivity is not clear. Purpose: To determine the effects of four weeks of NMES on insulin sensitivity, substrate utilization, and muscle mass in sedentary humans. Methods: Sedentary overweight/obese subjects (n=10; Age 36.8 ±3.8 yrs; BMI 32.0 ±1.3 kg/m2) were randomized into a control (Con) or NMES group. All participants received bilateral quadriceps stimulation (3x/wk; 30 min/ses) either at low intensity sensory level (Con) or high intensity neuromuscular level (NMES) for 4 weeks (50Hz). Insulin sensitivity was assessed by oral glucose tolerance test (OGTT), substrate utilization by indirect calorimetry, blood lactate by lactate analyzer, and body composition by DXA. Results: Con and NMES groups had comparable fasting blood glucose, glucose tolerance, substrate utilization, and muscle mass (all p>0.05) at baseline. Four weeks of NMES tended to improve glucose tolerance, measured by glucose area under the curve (AUC) (455.6 ±26.1 to 415.4 ±25.9 AU; p=0.07) whereas no change was observed in Con (430.7 ±20.2 to 494.7 ±77.2 AU; p=0.76). Glucose level after 2 hours of glucose consumption during OGTT was significantly lower after 4 weeks of NMES (150.9 ±7.6 to 138.2 ±7.6 mg/dL; p=0.03) whereas no change was observed in Con (151.8 ±6.1 to 173.2 ±34.2 mg/dL; p=0.33).Additionally, lactate AUC assessed during 30 min of NMES, was significantly greater compared to that of Con (p Conclusion: NMES is a novel and effective strategy to improve insulin sensitivity and glucose utilization in an at-risk overweight/obese sedentary population. Disclosure S. Bajpeyi: None. M. Galvan: None. M.J. Sanchez: None. J.B. Boyle: None. K. Min: None. F.J. Agullo: None. J.D. Covington: None. Funding University of Texas at El Paso

Multiple sclerosis (MS) is a metabolically demanding disease involving immune-mediated destruction of myelin in the central nervous system. We previously demonstrated a significant alteration in disease course in the experimental autoimmune encephalomyelitis (EAE) preclinical model of MS due to diet. Based on the established crosstalk between metabolism and gut microbiota, we took an unbiased sampling of microbiota, in the stool, and metabolites, in the serum and stool, from mice (Mus musculus) on the two different diets, the Teklad global soy protein-free extruded rodent diet (irradiated diet) and the Teklad sterilisable rodent diet (autoclaved diet). Within the microbiota, the genus Lactobacillus was found to be inversely correlated with EAE severity. Therapeutic treatment with Lactobacillus paracasei resulted in a significant reduction in the incidence of disease, clinical scores and the amount of weight loss in EAE mice. Within the metabolites, we identified shifts in glycolysis and the tricarboxylic acid cycle that may explain the differences in disease severity between the different diets in EAE. This work begins to elucidate the relationship between diet, microbiota and metabolism in the EAE preclinical model of MS and identifies targets for further study with the goal to more specifically probe the complex metabolic interaction at play in EAE that may have translational relevance to MS patients.

Background In the healthy central nervous system (CNS), microglia are found in a homeostatic state and peripheral macrophages are absent from the brain. Microglia play key roles in maintaining CNS homeostasis and acting as first responders to infection and inflammation, and peripheral macrophages infiltrate the CNS during neuroinflammation. Due to their distinct origins and functions, discrimination between these cell populations is essential to the comprehension of neuroinflammatory disorders. Studies comparing the gene profiles of microglia and peripheral macrophages, or macrophages in vitro-derived from bone marrow, under non-infectious conditions of the CNS, have revealed valuable microglial-specific genes. However, studies comparing gene profiles between CNS-infiltrating macrophages and microglia, when both are isolated from the CNS during viral-induced neuroinflammation, are lacking. Methods We isolated, via flow cytometry, microglia and infiltrating macrophages from the brains of Theiler’s murine encephalomyelitis virus-infected C57BL/6 J mice and used RNA-Seq, followed by validation with qPCR, to examine the differential transcriptional profiles of these cells. We utilized primary literature defining subcellular localization to determine whether or not particular proteins extracted from the transcriptional profiles were expressed at the cell surface. The surface expression and cellular specificity of triggering receptor expressed on myeloid cells 1 (TREM-1) protein were examined via flow cytometry. We also examined the immune response gene profile within the transcriptional profiles of these isolated microglia and infiltrating macrophages. Results We have identified and validated new microglial- and macrophage-specific genes, encoding cell surface proteins, expressed at the peak of neuroinflammation. TREM-1 protein was confirmed to be expressed by infiltrating macrophages, not microglia, at the peak of neuroinflammation. We also identified both unique and redundant immune functions, through examination of the immune response gene profiles, of microglia and infiltrating macrophages during neurotropic viral infection. Conclusions The differential expression of cell surface-specific genes during neuroinflammation can potentially be used to discriminate between microglia and macrophages as well as provide a resource that can be further utilized to target and manipulate specific cell responses during neuroinflammation. Electronic supplementary material The online version of this article (10.1186/s12974-019-1545-x) contains supplementary material, which is available to authorized users.

Supplemental Digital Content is available in the text., Objectives: Sepsis is a common cause of morbidity and mortality. A reliable, rapid, and early indicator can help improve efficiency of care and outcomes. To assess the IntelliSep test, a novel in vitro diagnostic that quantifies the state of immune activation by measuring the biophysical properties of leukocytes, as a rapid diagnostic for sepsis and a measure of severity of illness, as defined by Sequential Organ Failure Assessment and Acute Physiology and Chronic Health Evaluation-II scores and the need for hospitalization. Design, Setting, SUBJECTS: Adult patients presenting to two emergency departments in Baton Rouge, LA, with signs of infection (two of four systemic inflammatory response syndrome criteria, with at least one being aberration of temperature or WBC count) or suspicion of infection (a clinician order for culture of a body fluid), were prospectively enrolled. Sepsis status, per Sepsis-3 criteria, was determined through a 3-tiered retrospective and blinded adjudication process consisting of objective review, site-level clinician review, and final determination by independent physician adjudicators. MEASUREMENTS AND MAIN RESULTS: Of 266 patients in the final analysis, those with sepsis had higher IntelliSep Index (median = 6.9; interquartile range, 6.1–7.6) than those adjudicated as not septic (median = 4.7; interquartile range, 3.7–5.9; p < 0.001), with an area under the receiver operating characteristic curve of 0.89 and 0.83 when compared with unanimous and forced adjudication standards, respectively. Patients with higher IntelliSep Index had higher Sequential Organ Failure Assessment (3 [interquartile range, 1–5] vs 1 [interquartile range, 0–2]; p < 0.001) and Acute Physiology and Chronic Health Evaluation-II (7 [interquartile range, 3.5–11.5] vs 5 [interquartile range, 2–9]; p < 0.05) and were more likely to be admitted to the hospital (83.6% vs 48.3%; p < 0.001) compared with those with lower IntelliSep Index. CONCLUSIONS: In patients presenting to the emergency department with signs or suspicion of infection, the IntelliSep Index is a promising tool for the rapid diagnosis and risk stratification for sepsis.

GNAQ and GNA11 (GNAQ/11) mutations are found in less than 2% of all melanoma, but more than 80% of uveal melanoma. Mutations in these Gα proteins lead to constitutive activation of multiple oncogenic pathways, including MAPK (RAF->MEK1/2->ERK1/2) and YAP signaling. Metastatic uveal melanoma is refractory to all forms of pharmacologic treatment, such as FDA-approved targeted therapies inhibiting MEK1/2 (i.e. trametinib and binimetinib). We show that combining MEK1/2 inhibitors with 4-aminoquinoline antimalarials, chloroquine or hydroxychloroquine, resulted in synergistic and apoptosis-mediated cytotoxicity in GNAQ/11 mutant uveal melanoma cell lines. Interestingly, in contrast to our previous work in pancreatic and other RAS-driven cancers, the lysosomotropic role of chloroquine was not sufficient to promote cytotoxicity with MEK1/2 inhibitors, as neither lysosome inhibition with Bafilomycin A1 nor autophagy-specific and macropinocytosis-specific inhibition yielded enhanced cell death in combination with MEK1/2 inhibition. We then found that chloroquine prevented nuclear localization of the transcriptional coactivator, YAP, suggesting a novel mechanism of chloroquine. YAP inhibition combined with MEK1/2 inhibition enhanced cell death only in the presence of Bafilomycin A1. Gα-specific inhibition (inhibiting YAP and MAPK) combined with Bafilomycin A1 yielded similar results. This implies that the ability of chloroquine to inhibit both YAP signaling and lysosome function is required for promoting cell death in the presence of MEK1/2 inhibition. For in vivostudies, we utilized a hepatic colonization model using luciferized human metastatic uveal melanoma cell lines, OMM2.5 and OMM1. Daily treatment of trametinib with hydroxychloroquine in combination resulted in delayed tumor growth and increased overall survival compared to either treatment as monotherapy or chemotherapy. These findings were also recapitulated in an immunocompetent mouse model in which immortalized mouse melanocytes (Melan-A) with either a GNAQ or GNA11 activating mutation were implanted into syngeneic C57BL/6 mice. Our findings identify a novel mechanism of chloroquine and suggest a potentially effective strategy combining two FDA-approved drugs for the treatment of metastatic uveal melanoma. Citation Format: Amanda Truong, Michael Scherzer, Conan Kinsey, John Michael Sanchez, Jae Hyuk Yoo, Jackson Richards, Donghan Shin, Phaedra Ghazi, Michael Onken, Kendall Blumer, Shannon Odelberg, Martin McMahon. Chloroquine synergizes with MEK1/2 targeted therapy through dual YAP and lysosomal inhibition in GNAQ/11 mutant uveal melanoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1890.

Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system (CNS) that affects 2.5 million people worldwide. Growing evidence suggests that perturbation of the gut microbiota, a dense collection of microorganisms that colonize the gut, plays a functional role in MS. Utilizing experimental autoimmune encephalomyelitis (EAE), the most commonly used murine model of MS, we previously identified a significant negative correlation between the relative abundance of gut Lactobacillus and the severity of EAE, mirroring what is seen in MS patients. We now report that a human gut-derived commensal strain of Lactobacillus paracasei (Lb) can suppress preclinical murine models of MS with both prophylactic and therapeutic administration. Surprisingly, we found that heat-killed Lb was as effective as live Lb in protecting animals against EAE, as measured by disease score and demyelination. Heat-killed Lb-treated animals had reduced numbers of CNS-infiltrating leukocytes and this was associated with a decrease in select chemokines in the serum. The beneficial effect of heat-killed Lb was dependent on host expression of Toll-like receptor 2 (TLR2), which senses major components of the Lb cell wall, as EAE is not suppressed by heat-killed Lb in TLR2-deficient mice. Thus, Lb-associated molecular patterns may be an important signal to prevent aberrant peripheral immune cell infiltration into the CNS and warrant further investigation as a novel therapy for MS patients.

Inter-ethnic violence has plagued many areas of the globe, but not many regions have been impacted by the level of violence and atrocity that has been documented during the conflicts following the collapse of the Yugoslavian Republic. In particular, the clash between Serbian Christians and Muslim Albanians in the Serbian region of Kosovo resulted in international military intervention and the rare incidence of United Nations governance in that previously unsettled area. As a direct result, a fully functional criminal justice sector and ethnically balanced Kosovo Police now serve the region. But these efforts have not come without regional geo-political differences and disagreement. The recent initiatives undertaken by the Kosovo government and police to enhance inter-ethnic relations have been outlined in detail.

Microglia are the only resident myeloid cell in the central nervous system (CNS) parenchyma, but the role of microglia in the context of neurotropic viral infection is poorly understood. Using different amounts of Theiler's murine encephalomyelitis virus (TMEV) in a preclinical model of epilepsy and PLX5622, a colony stimulating factor-1 receptor inhibitor that selectively depletes microglia in the CNS, we report that microglia-depleted, TMEV-infected mice develop seizures, manifest paralysis, and uniformly succumb to fatal encephalitis regardless of viral amount. CNS demyelination correlates with viral amount; however, viral amount does not correlate with axon damage and TMEV antigen in the CNS.

Malpelo es Colombia, Maravilla Estrategica esta compuesto por siete capitulos. Comenzamos nuestro viaje a traves del reconocimiento de los diferentes esfuerzos que se han realizado para que Malpelo sea conocida como un punto estrategico de Colombia. Luego, en el segundo capitulo, empezamos a comprender la complejidad de Malpelo. Su lejania, sus ambientes extremos, la intensa interaccion entre la roca y las olas, sus inclinadas pendientes, y las condiciones abioticas de sus aguas, que hacen que este lugar albergue abundantes individuos de especies particulares. Estas descripciones fisicas nos abren el camino para indagar en su diversidad. Es asi como en el tercer capitulo nos sumergimos en sus aguas para conocer pequenos organismos esenciales en la cadena trofica y en la dinamica de las poblaciones del entorno de la isla; tambien, los fundadores exitosos de los ecosistemas marinos, como las esponjas, los corales que, increiblemente logran conquistar mas de 30 m de profundidad; los briozoos con sus novedosos descubrimientos, los moluscos, artropodos, equinodermos y otros invertebrados con su alta diversidad y conspicuidad, para terminar nadando con los grandes vertebrados, los peces y mamiferos que sorprenden a los cientificos con sus interacciones, endemismos, abundancias y comportamientos. Seguido, escalamos las paredes rocosas para descubrir, en el capitulo cuarto, la vida terrestre de la isla, que presenta una diversidad muy singular con una interaccion tan estrecha, agresiva, explosiva y subita que dejara sorprendidos a los lectores. Para concluir con las caracteristicas biologicas de Malpelo, en el quinto se expone el conocimiento de la diversidad de la flora, una diversidad que en su parte terrestre es bastante escasa, y en la marina desconocida, sin dejar a un lado su importancia en la dinamica de los ecosistemas. La ultima seccion del libro es un llamado a mejorar los usos y el control en el Santuario. Es asi como en el capitulo sexto se describe una de las mayores amenazas a la diversidad y las oportunidades de aprovechamiento turistico del santuario. Para concluir, en el septimo se exponen las proyecciones de Malpelo, para mejorar su conocimiento cientifico, gestion, proteccion y apropiacion en el futuro.

Widespread prevalence of multidrug and pandrug-resistant bacteria has prompted substantial concern over the global dissemination of antibiotic resistance genes (ARGs). Environmental compartments can behave as genetic reservoirs and hotspots, wherein resistance genes can accumulate and be laterally transferred to clinically relevant pathogens. In this work, we explore the ARG copy quantities in three environmental media distributed across four cities in California and demonstrate that there exist city-to-city disparities in soil and drinking water ARGs. Statistically significant differences in ARGs were identified in soil, where differences in blaSHV gene copies were the most striking; the highest copy numbers were observed in Bakersfield (6.0 × 10–2 copies/16S-rRNA gene copies and 2.6 × 106 copies/g of soil), followed by San Diego (1.8 × 10–3 copies/16S-rRNA gene copies and 3.0 × 104 copies/g of soil), Fresno (1.8 × 10–5 copies/16S-rRNA gene copies and 8.5 × 102 copies/g of soil), and Los Angeles (5.8 × 10–6 copies/16S-rRNA gene copies and 5.6 × 102 copies/g of soil). In addition, ARG copy numbers in the air, water, and soil of each city are contextualized in relation to globally reported quantities and illustrate that individual genes are not necessarily predictors for the environmental resistome as a whole.

Background The traditional intraoral manual reduction of temporomandibular joint (TMJ) dislocations is time consuming, difficult, and at times ineffective, and commonly requires conscious sedation. Objectives We describe a novel technique for the reduction of acute nontraumatic TMJ dislocations in the emergency department (ED). Methods This study was a prospective convenience sample population during a 3-year period at two university teaching-hospital EDs where acute nontraumatic TMJ dislocations were reduced utilizing our syringe technique. Demographics, mechanism, duration of dislocation, and reduction time were collected. Briefly, the “syringe” technique is a hands-free technique that requires a syringe to be placed between the posterior molars as they slide over the syringe to glide the anteriorly displaced condyle back into its normal anatomical position. Procedural sedation or intravenous analgesia is not required. Results Of the 31 patients, the mean age was 38 years. Thirty patients had a successful reduction (97%). The majority of dislocations were reduced in Conclusion We describe a novel technique for the reduction of the acutely nontraumatic TMJ dislocation in the ED. It is simple, fast, safe, and effective.