Concurrent Activation Research Articles

Abstract 658: AKT pathway as a therapeutic target to constrain lineage plasticity leading to histological transdifferentiation

Abstract Lineage plasticity contributes to therapeutic resistance in cancer. In lung adenocarcinomas (LUADs), this phenomenon drives neuroendocrine (NE) and squamous cell (LUSC) histologic transdifferentiation in the context of acquired resistance to targeted inhibition of driver mutations, with up to 14% and 9% incidences in EGFR-mutant tumors relapsed on EGFR inhibitors, respectively. Notably, survival of patients with NE- or LUSC-transdifferentiated tumors is lower than that of either LUAD or de novo LUSC patients. To date, little is known about the molecular effectors enhancing lineage plasticity and driving histological transdifferentiation due to the paucity of well annotated pre- and post-transdifferentiation clinical samples amenable for molecular analyses. Currently no specific therapies for LUSC or NE transdifferentiation prevention are available for patients at high risk of transformation. We performed multi-omic profiling of transdifferentiating clinical samples, as well as control never-transformed LUAD and de novo LUSC and small cell carcinomas, including comprehensive and integrative genomic (whole exome sequencing), epigenomic (bisulfite sequencing), transcriptomic (RNAseq) and protein (antibody arrays) characterization. Findings were validated in preclinical models including cell lines as well as LUSC- and NE-transdifferentiation patient-derived xenograft models. Our data suggest that histological transdifferentiation is driven by epigenetic -rather than mutational- events, and indicate that transdifferentiated tumors retain molecular features of their previous LUAD state. Integrative analysis revealed biological pathways dysregulated specifically for distinct histological outcomes, including downregulation of RTK signaling and Notch-related genes in NE-transformed tumors, and upregulation of genes involved in Hedgehog and Notch signaling and MYC targets in LUSC-transdifferentiated tumors. Most interestingly, these analyses revealed commonly dysregulated pathways for transdifferentiated tumors, including marked downregulation of a variety of immune-related pathways and upregulation of genes involved in AKT signaling and in the PRC2 epigenetic remodeling complex. Concurrent activation of AKT and MYC overexpression induced a squamous phenotype in EGFR-mutant LUAD preclinical models, further accentuated by EGFR inhibition. Pharmacological targeting of AKT in combination with osimertinib delayed both squamous and NE transformation in EGFR-mutant patient-derived xenograft transdifferentiation models. These results identify common and histology-specific drivers and dysregulated pathways in NE and LUSC transdifferentiation, and nominate AKT as a therapeutic target to constrain lineage plasticity and prevent the acquisition of resistance to EGFR-targeted therapies through histological transdifferentiation. Citation Format: Alvaro Quintanal-Villalonga, Hirokazu Taniguchi, Yingqian A. Zhan, Fathema Uddin, Viola Allaj, Parvathy Manoj, Nisargbhai S. Shah, Umesh K. Bhanot, Jacklynn Egger, Juan Qiu, Elisa de Stanchina, Natasha Rekhtman, Brian Houck-Loomis, Richard P. Koche, Helena A. Yu, Triparna Sen, Charles M. Rudin. AKT pathway as a therapeutic target to constrain lineage plasticity leading to histological transdifferentiation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 658.

Evaluation of signal analysis algorithms for ipsilateral motor-evoked potentials induced by transcranial magnetic stimulation

Objective. Evaluating ipsilateral motor-evoked potentials (iMEP) induced by transcranial magnetic stimulation is challenging. In healthy adults, isometric contraction is necessary to facilitate iMEP induction; therefore, the signal may be masked by the concurrent muscle activity. Signal analysis algorithms for iMEP evaluation need to be benchmarked and evaluated. Approach. An open analysis toolbox for iMEP evaluation was implemented on the basis of 11 previously reported algorithms, which were all threshold based, and a new template-based method based on data-driven signal decomposition. The reliability and validity of these algorithms were evaluated with a dataset of 4244 iMEP from 55 healthy adults. Main results. iMEP estimation varies drastically between algorithms. Several algorithms exhibit high reliability, but some appear to be influenced by background activity of muscle preactivation. Especially in healthy subjects, template-based approaches might be more valid than threshold-based ones. Measurement of iMEP persistence requires algorithms that reject some trials as MEP negative. The stricter the algorithms reject trials, the less reliable they generally are. Our evaluation identifies an optimally strict and reliable algorithm. Significance. We show different benchmarks and propose application for different use cases.

Distribution and Morphology of Lava Tube Systems on the Western Flank of Alba Mons, Mars

AbstractIntegrated analyses of Thermal Emission Imaging System (THEMIS) IR, Context Camera (CTX), and Mars Orbiter Laser Altimeter (MOLA) data sets have been used to characterize the western flank of the Martian volcano Alba Mons, which hosts a prominent population of lava tube systems. Identification and mapping of lava tube systems is based on both morphologic and topographic analyses, including the presence of chains of collapse depressions and elongate, sinuous ridges. Lava tubes and adjacent tabular lava flows with lengths of 100+ km form an extensive lava flow field. Analyses of topographic data sets, including slope maps, suggest continuity of the radial flow field pattern across the full western flank. Concurrent surface activity across the western flank is consistent with age constraints from geologic mapping and crater size‐frequency distributions that indicate Early Amazonian ages. The mapped population of 331 lava tube systems in the western flank geologic map quadrangle has a mean length of 36.2 km and a total length of ∼12,000 km. Individual lava tube systems extend up to ∼400 km. Orientation and slope data for lava tube systems show small deviations compared to regional values in 50‐km grid cells defined by the MOLA Digital Elevation Model, suggesting a strong coupling of lava tubes to the current slopes of Alba Mons. Mapping of lava tube systems documents segments both showing collapse and with no collapse, indicating the potential for extensive subsurface cavities that would be important astrobiological targets.

AKT inhibition as a therapeutic strategy to constrain histological transdifferentiation in EGFR-mutant lung adenocarcinoma.

e21166 Background: In lung adenocarcinomas (LUADs), lineage plasticity drives neuroendocrine (NE) and squamous cell (LUSC) transdifferentiation in the context of acquired resistance to targeted inhibition of driver mutations, with up to 14% and 9% incidences in EGFR-mutant tumors relapsed on EGFR inhibitors, respectively. Notably, survival of patients with NE- or LUSC-transdifferentiated tumors is remarkably lower than those of LUAD or de novo LUSC patients. The paucity of transforming clinical specimens amenable for molecular analyses has hindered the identification of histological transformation drivers, and to date no specific therapies aimed to prevent or delay transdifferentiation-led therapy relapse are available for patients at high risk of transformation. Methods: We performed multi-omic profiling of LUAD-to-LUSC and LUAD-to-NE transdifferentiating clinical samples, including comprehensive and integrative genomic (whole exome sequencing), epigenomic (bisulfite sequencing), transcriptomic (RNAseq) and protein (antibody arrays) characterization. Clinical findings were validated in preclinical models including cell lines as well as LUSC- and NE-transdifferentiation patient-derived xenograft models. Results: Our data supports that histological transdifferentiation from LUAD to LUSC or NE tumors is driven by epigenetic remodeling rather than by mutational events, and indicate that transdifferentiated tumors retain epigenomic features of their previous LUAD state. Integrative epigenomic, transcriptomic and protein analysis revealed divergent biological pathways dysregulated for each histological outcome, such as downregulation of RTK signaling and Notch-related genes in NE-transformed tumors, and upregulation of genes involved in Hedgehog and Notch signaling and MYC targets in LUSC-transdifferentiated tumors. Most interestingly, these analyses identified commonly dysregulated pathways in both NE- and LUSC-transdifferentiating tumors, including remarkable downregulation of a variety of immune-related pathways and upregulation of genes involved in AKT signaling and in the PRC2 epigenetic remodeling complex. Concurrent activation of AKT and MYC overexpression induced a squamous phenotype in EGFR-mutant LUAD preclinical models, further accentuated by EGFR inhibition. Pharmacological targeting of AKT in combination with osimertinib delayed both squamous and NE transformation in different EGFR-mutant patient-derived xenograft transdifferentiation models. Conclusions: These results identify common and divergent dysregulated pathways in NE and LUSC transdifferentiation, and nominate AKT as a therapeutic target to prevent the acquisition of resistance to EGFR-targeted therapies through histological transdifferentiation.

Urine ALCAM, PF4 and VCAM-1 Surpass Conventional Metrics in Identifying Nephritis Disease Activity in Childhood-Onset Systemic Lupus Erythematosus.

ObjectivesSerial kidney biopsy for repeat evaluation and monitoring of lupus nephritis (LN) in childhood-onset Systemic Lupus Erythematosus (cSLE) remains challenging, thus non-invasive biomarkers are needed. Here, we evaluate the performance of ten urine protein markers of diverse nature including cytokines, chemokines, and adhesion molecules in distinguishing disease activity in cSLE.MethodsEighty-four pediatric patients meeting ≥4 ACR criteria for SLE were prospectively enrolled for urine assay of 10 protein markers normalized to urine creatinine, namely ALCAM, cystatin-C, hemopexin, KIM-1, MCP-1, NGAL, PF-4, Timp-1, TWEAK, and VCAM-1 by ELISA. Samples from active renal (LN) and active non-renal SLE patients were obtained prior to onset/escalation of immunosuppression. SLE disease activity was evaluated using SLEDAI-2000. 59 patients had clinically-active SLE (SLEDAI score ≥4 or having a flare), of whom 29 patients (34.5%) were classified as active renal, and 30 patients (35.7%) were active non-renal. Twenty-five healthy subjects were recruited as controls.ResultsUrine concentrations of ALCAM, KIM-1, PF4 and VCAM-1 were significantly increased in active LN patients versus active non-renal SLE, inactive SLE and healthy controls. Five urine proteins differed significantly between 2 (hemopexin, NGAL, MCP1) or 3 (Cystatin-C, TWEAK) groups only, with the highest levels detected in active LN patients. Urine ALCAM, VCAM-1, PF4 and hemopexin correlated best with total SLEDAI as well as renal-SLEDAI scores (p < 0.05). Urine ALCAM, VCAM-1 and hemopexin outperformed conventional laboratory measures (anti-dsDNA, complement C3 and C4) in identifying concurrent SLE disease activity among patients (AUCs 0.75, 0.81, 0.81 respectively), while urine ALCAM, VCAM-1 and PF4 were the best discriminators of renal disease activity in cSLE (AUCs 0.83, 0.88, 0.78 respectively), surpassing conventional biomarkers, including proteinuria. Unsupervised Bayesian network analysis based on conditional probabilities re-affirmed urine ALCAM as being most predictive of active LN in cSLE patients.ConclusionUrinary ALCAM, PF4, and VCAM-1 are potential biomarkers for predicting kidney disease activity in cSLE and hold potential as surrogate markers of nephritis flares in these patients.

Knowledge of patients with type II diabetes mellitus regarding preventive measures of foot ulcer

A foot ulcer is a dangerous diabetic problem caused by the concurrent physiological activities of ischemia, neuropathy, and infection in patients with type II diabetes mellitus. However, patients' knowledge about foot care is essential for avoiding foot ulceration.A cross-sectional descriptive design is carried out in the current study from September 9th 2021 to May 9th 2022, in order to assess the levels of knowledge of type II diabetic patients regarding preventive measures of foot ulcers. A non-probability (purposive sample) of 300 diabetic patients type II. Data collected using a well-designed questionnaire consists of three parts: Part I consists of demographic data. Part II consists of Clinical Data. Part III includes Patients' knowledge regarding preventive measures for foot ulcers. The findings indicate there is a significant association between patients' knowledge and (educational status and occupation). Also, there was a significant association between patients' knowledge and (duration of the disease and education about diabetes). The study concluded that patients with type II diabetes have moderate knowledge about preventive measures for foot ulcers.

Efektivitas Pembelajaran Daring Di Sekolah MTSN 1 Murung

The purpose of this study is to find out the effectiveness of online learning in MTsN 1 Murung. The questions you want to answer through this study are: 1) To find out the effectiveness of online learning in MTsN 1 Moody. 2) To find out the advantages and disadvantages in the implementation of online learning in MTsN 1 Murung. 3) To know what needs to be improved so that the implementation of online learning runs better and in accordance with the wishes. This research uses qualitative research methods with qualitative descriptive analysis techniques or interview data collection and documentation. The subjects of the study were the head of the madrassa, teachers and learners of MTsN 1 Murung. The data analysis technique in this study uses qualitative descriptive data analysis of interactive models from Milles and Michael Huberman consisting of three concurrent activity paths, namely; data reduction, presentation of data, and withdrawal of conclusions. Based on the results of research and interviews showed that online learning in MTsN 1 Murung is not effective considering that in its short implementation so that gutu can not know whether learners have understood the material or not, the advantages of online learning in MTsN 1 Murung Raya that learning can be accessed anywhere and anytime, learners can learn independently, the provision of tasks is more structured through applications. While the lack of online learning in MTsN 1 Murung is monitoring or monitoring the situation of students sulik done, signals / connections that are less stable in some areas. The field that needs to be improved so that online learning can run well is to increase the creativity of teachers in making learning media that are easy to understand, monitoring / supervision to learners is more tightened, and cooperation between parents and teachers must be well established

18β-Glycyrrhetinic Acid Protects against Cholestatic Liver Injury in Bile Duct-Ligated Rats.

18β-Glycyrrhetinic acid is a nutraceutical agent with promising hepatoprotective effects. Its protective mechanisms against cholestatic liver injury were further investigated in a rodent model of extrahepatic cholestasis caused by Bile Duct Ligation (BDL) in rats. The daily oral administration of 18β-Glycyrrhetinic acid improved liver histology, serum biochemicals, ductular reaction, oxidative stress, inflammation, apoptosis, impaired autophagy, and fibrosis. 18β-Glycyrrhetinic acid alleviated the BDL-induced hepatic and systemic retention of bile acids, matrix-producing cell activation, hepatic collagen deposition, Transforming Growth Factor beta-1/Smad activation, malondialdehyde elevation, glutathione reduction, High Mobility Group Box-1/Toll-Like Receptor-4 activation, NF-κB activation, inflammatory cell infiltration/accumulation, Interleukin-1β expression, Signal Transducer and Activator of Transcription-1 activation, Endoplasmic Reticulum stress, impairment autophagy, and caspase 3 activation. Conversely, the protein expression of Sirt1, Farnesoid X Receptor, nuclear NF-E2-Related Factor-2, Transcription Factor EB, bile acid efflux transporters, and LC3-II, as well as the protein phosphorylation of AMP-Activated Protein Kinase, was promoted in 18β-Glycyrrhetinic acid-treated BDL rats. The hepatoprotective effects of 18β-Glycyrrhetinic acid in the present investigation correlated well with co-activation and possible interactions among Sirt, FXR, and Nrf2. The concurrent or concomitant activation of Sirt1, FXR, and Nrf2 not only restored the homeostatic regulation of bile acid metabolism, but also alleviated oxidative stress, inflammation, apoptosis, impaired autophagy, and fibrosis.

Effects of 8-week oral splint usage on body flexibility and muscle strength-endurance performance in Pilates practitioners

PurposeWe investigated the acute and chronic effects of oral splint on muscle strength-endurance performance and body flexibility.MethodsTwelve Pilates practitioners (age 49.5 ± 5.8 years; mass 58.6 ± 6.5 kg) were evaluated with (OS) and without (CTRL) oral splint. Before (PRE) and after (POST) 8 weeks of oral splint usage during Pilates activity, muscle strength-endurance and body flexibility were evaluated through a frontal plank exercise and a Toe-touch test in OS and CTRL. Before each test, the highest electrical activation (EMG) of masseter and temporal muscles during maximal occlusion was determined. During frontal plank, the time-to-exhaustion, the maximum voluntary activation of masticatory muscles and the time of muscles activation have been determined.ResultsNo OS vs CTRL and PRE vs POST differences were found in the maximum voluntary activation of temporal and masseter muscles. The time-to-exhaustion in OS was longer than CTRL in both PRE (P = 0.049) and POST sessions (P = 0.043). Time of masticatory muscles activation during plank was moderately greater in OS at PRE (P = 0.020) and POST (P = 0.022), while no difference between PRE and POST emerged in both conditions. Higher muscular activation was found in OS than CTRL at PRE (P = 0.031) and POST (P = 0.031), whereas no difference between PRE and POST appeared. No effect on flexibility emerged neither acutely nor chronically.ConclusionAcute and chronic oral splint usage improved strength-endurance performance but not body flexibility. These findings suggest biomechanical and neural mechanisms influencing the muscle length and the concurrent activation potentiation, but not muscle tone and stiffness.

The association of momentary sad moods, concurrent productive behaviour, and global functional outcomes: a 30-day ecological momentary assessment study of people with bipolar illness

ABSTRACT Background Previous weekly sampling studies found that persistent sad moods are associated with disability in bipolar illness. However, those data were collected retrospectively. We examined the momentary quality of activities (productive, unproductive, and passive recreation) in an ecological momentary assessment (EMA) study and related sadness at each survey to quality of momentary activities and overall everyday functioning. Methods Participants with bipolar illness (N = 91) were sampled three times per day for 30 days. Each survey queried participants as to where they were, with whom, what they were doing, and their mood state. Activities were characterised according to predetermined criteria and related to momentary sadness. Observer ratings of everyday functioning were related to daily reports of sadness and activities. Results Sadness was associated with the quality of activities. Momentary reports of unproductive activities were associated with the most sadness (p < .001), followed by passive recreation, and productive activities. Momentary sadness and momentary unproductive activities correlated with observer ratings of competence in work, everyday activities, and social outcomes (p < .001). Using both predictors led to the best model. Conclusions This study on the course of sad moods in people with bipolar illness to EMA found that momentary sadness correlatesdwith the quality of concurrent activities and that both sadness and the quality of everyday activities predicted observer ratings of everyday functioning. Although we cannot determine the causal direction, these findings support the hypothesis that momentary sadness leads to reductions in productive activities and impairments in everyday functioning.

A light-induced small G-protein gem limits the circadian clock phase-shift magnitude by inhibiting voltage-dependent calcium channels.

Calcium signaling is pivotal to the circadian clockwork in the suprachiasmatic nucleus (SCN), particularly in rhythm entrainment to environmental light-dark cycles. Here, we show that a small G-protein Gem, an endogenous inhibitor of high-voltage-activated voltage-dependent calcium channels (VDCCs), is rapidly induced by light in SCN neurons via the calcium (Ca2+)-mediated CREB/CRE transcriptional pathway. Gem attenuates light-induced calcium signaling through its interaction with VDCCs. The phase-shift magnitude of locomotor activity rhythms by light, at night, increases in Gem-deficient (Gem-/-) mice. Similarly, in SCN slices from Gem-/- mice, depolarizing stimuli induce larger phase shifts of clock gene transcription rhythms that are normalized by the application of an L-type VDCC blocker, nifedipine. Voltage-clamp recordings from SCN neurons reveal that Ca2+ currents through L-type channels increase in Gem-/- mice. Our findings suggest that transcriptionally activated Gem feeds back to suppress excessive light-evoked L-type VDCC activation, adjusting the light-induced phase-shift magnitude to an appropriate level in mammals.

A tumor-targeted cytokine/antibody fusion protein to stimulate anti-cancer immunity

Abstract Interleukin-2 (IL-2) is a pleiotropic cytokine that has clinical utility in stimulating anti-cancer immunity. However, the clinical efficacy of IL-2 is limited by its short serum half-life, systemic toxicity, and concurrent activation of immunostimulatory natural killer (NK) and effector T (Teff) cells and immunosuppressive regulatory T (Treg) cells. IL-2 can signal down an intermediate-affinity receptor consisting of IL-2 receptor beta (IL-2Rβ) and common gamma chain (γc) or a high-affinity receptor that also includes IL-2Rα. Tregs express high levels of IL-2Rα, making them much more responsive to IL-2 than naïve NK and Teff cells. To extend the serum half-life and selectively bias the immunostimulatory functions of IL-2 in a stable unimolecular format, we created a single-chain fusion protein, denoted F10 immunocytokine (F10 IC), that intramolecularly fuses human IL-2 to an anti-IL-2 antibody that sterically blocks the IL-2/IL-2Rα interaction and allosterically enhances the IL-2/IL-2Rβ interaction. To exploit the abundance and accessibility of collagen in the tumor microenvironment for targeting, we fused collagen binding domains to F10 IC in various topographies. These novel fusion proteins occlude IL-2Rα binding and enhance IL-2Rβ binding, thereby eliminating the Treg advantage conferred by IL-2Rα and preferentially activating immune effector cells. Furthermore, these proteins selectively bind purified collagen and are capable of homing to, and being retained in, the tumor in vivo. Building on these promising results, we are assessing the in vivo therapeutic efficacy of these fusion proteins in various murine tumor models to illustrate the potential of collagen binding F10 IC as a powerful anti-cancer therapeutic. Supported by a Sanofi Innovation Award and NIH grants (R01 EB029341, T32 AI007417)

Validation of an Automated Wearable Camera-Based Image-Assisted Recall Method and the 24-h Recall Method for Assessing Women's Time Allocation in a Nutritionally Vulnerable Population: The Case of Rural Uganda.

Accurate data are essential for investigating relationships between maternal time-use patterns and nutritional outcomes. The 24 h recall (24HR) has traditionally been used to collect time-use data, however, automated wearable cameras (AWCs) with an image-assisted recall (IAR) may reduce recall bias. This study aimed to evaluate their concurrent criterion validity for assessing women’s time use in rural Eastern Ugandan. Women’s (n = 211) time allocations estimated via the AWC-IAR and 24HR methods were compared with direct observation (criterion method) using the Bland–Altman limits of agreement (LOA) method of analysis and Cronbach’s coefficient alpha (time allocation) or Cohen’s κ (concurrent activities). Systematic bias varied from 1 min (domestic chores) to 226 min (caregiving) for 24HR and 1 min (own production) to 109 min (socializing) for AWC-IAR. The LOAs were within 2 h for employment, own production, and self-care for 24HR and AWC-IAR but exceeded 11 h (24HR) and 9 h (AWC-IAR) for caregiving and socializing. The LOAs were within four concurrent activities for 24HR (−1.1 to 3.7) and AWC-IAR (−3.2 to 3.2). Cronbach’s alpha for time allocation ranged from 0.1728 (socializing) to 0.8056 (own production) for 24HR and 0.2270 (socializing) to 0.7938 (own production) for AWC-IAR. For assessing women’s time allocations at the population level, the 24HR and AWC-IAR methods are accurate and reliable for employment, own production, and domestic chores but poor for caregiving and socializing. The results of this study suggest the need to revisit previously published research investigating the associations between women’s time allocations and nutrition outcomes.

Co-targeting B-RAF and PTEN Enables Sensory Axons to Regenerate Across and Beyond the Spinal Cord Injury.

Primary sensory axons in adult mammals fail to regenerate after spinal cord injury (SCI), in part due to insufficient intrinsic growth potential. Robustly boosting their growth potential continues to be a challenge. Previously, we showed that constitutive activation of B-RAF (rapidly accelerated fibrosarcoma kinase) markedly promotes axon regeneration after dorsal root and optic nerve injuries. The regrowth is further augmented by supplemental deletion of PTEN (phosphatase and tensin homolog). Here, we examined whether concurrent B-RAF activation and PTEN deletion promotes dorsal column axon regeneration after SCI. Remarkably, genetically targeting B-RAF and PTEN selectively in DRG neurons of adult mice enables many DC axons to enter, cross, and grow beyond the lesion site after SCI; some axons reach ∼2 mm rostral to the lesion by 3 weeks post-injury. Co-targeting B-RAF and PTEN promotes more robust DC regeneration than a pre-conditioning lesion, which additively enhances the regeneration triggered by B-RAF/PTEN. We also found that post-injury targeting of B-RAF and PTEN enhances DC axon regeneration. These results demonstrate that co-targeting B-RAF and PTEN effectively enhances the intrinsic growth potential of DC axons after SCI and therefore may help to develop a novel strategy to promote robust long-distance regeneration of primary sensory axons.

In a Degraded Stream, Increased Salt Concentration is Correlated with Decreased EPT Diversity

Many streams in the Northern Panhandle of West Virginia are biologically impaired due to historical and concurrent anthropogenic activities, including mining, agriculture, and urbanization. These activities alter water chemistry which affects the benthic macroinvertebrate communities. While taxa like Ephemeroptera, Plecoptera, and Trichoptera (EPT taxa) are sensitive to water pollution, other taxa like tipulids, beetles, and chironomid midges are more pollution tolerant. However, responses to specific chemical pollutants are taxa specific and even some EPT are relatively pollution tolerant. We investigated how increases in chloride (Cl) and specific conductance (SPC) affected the macroinvertebrate community in a stream already heavily impacted by mining, urbanization and agriculture. Five upstream sampling sites were placed in a degraded stream above the outfall of an active coal mine, while five additional sites were placed downstream of the discharge. Water chemistry was measured in each site, and macroinvertebrates were collected with kick nets, leaf packs and a visual collection of unique habitats following the Stream Monitoring Information Exchange (SMIE) protocol. We found that temperature, SPC, and Cl were significantly higher downstream of the mine outfall with SPC and Cl more than doubling. SMIE water quality scores and whole community Shannon-Wiener Diversity Index scores were not significantly different between upstream and downstream sites but EPT diversity was significantly higher upstream of the mine. This data helps determine taxa specific responses to changes in water chemistry, specifically SPC and Cl, and highlights how streams already afflicted with poor water quality can lose additional taxa when the concentrations of salts increases.

Individual and Contextual Risk and Protective Factors for Suicidal Thoughts and Behaviors among Black Adolescents with Arrest Histories.

Black adolescents in the United States have experienced an increase in suicidal thoughts and behaviors (STBs). Since Black adolescents are overrepresented in the youth punishment system, more research is needed to investigate correlates of STBs for this population. The purpose of this paper is to explore and establish correlates of individual, family, and community risk and protective factors and their relationship to lifetime STBs in a national sample of Black youth with arrest histories. Guided by an intersectional eco-behavioral lens, we investigated individual, family and contextual risk and protective factors for STBs among a national sample of justice-involved Black youth aged 12–17 with a history of arrest (n = 513). We used logistic regression models to test risk and protective factors for STBs. Among the sample, 9.78% endorsed suicidal ideation, and 7.17% endorsed a previous suicide attempt. Further, gender (female) and depression severity were risk factors for STBs, while positive parenting and religiosity were protective factors for STBs. School engagement was associated with lower levels of suicidal ideation. The findings suggest suicide prevention and intervention efforts should identify developmentally salient risk and protective factors to reduce mental health burden associated with STBs and concurrent alleged law-breaking activity of Black youth.

Circulating corticosterone levels mediate the relationship between acute ethanol intoxication and markers of NF-κB activation in male rats

Binge drinking is a harmful pattern of alcohol use that is associated with a number of serious health problems. Of particular interest are the rapid alterations in neuroimmune gene expression and the concurrent activation of the hypothalamic-pituitary-adrenal (HPA) axis activation associated with high intensity drinking. Using a rat model of acute binge-like ethanol exposure, the present studies were designed to assess the role of corticosterone (CORT) in ethanol-induced neuroimmune gene expression changes, particularly those associated with the NFκB signaling pathway, including rapid induction of IL-6 and IκBα, and suppression of IL-1β and TNFα gene expression evident after administration of moderate to high doses of ethanol (1.5–3.5 g/kg ip) during intoxication (3 h post-injection). Experiment 1 tested whether inhibition of CORT synthesis with metyrapone and aminoglutethimide (100 mg/kg each, sc) would block ethanol-induced changes in neuroimmune gene expression. Results indicated that rapid alterations in IκBα, IL-1β, and TNFα expression were completely blocked by pretreatment with the glucocorticoid synthesis inhibitors, an effect that was reinstated by co-administration of exogenous CORT (3.75 mg/kg) in Experiment 2. Experiment 3 assessed whether these rapid alterations in neuroimmune gene expression would be evident when rats were challenged with a subthreshold dose of ethanol (1.5 g/kg) in combination with 2.5 mg/kg CORT, which showed limited evidence for additive effects of low-dose CORT combined with a moderate dose of ethanol. Acute inhibition of mineralocorticoid (spironolactone) or glucocorticoid (mifepristone) receptors, alone (Experiment 4) or combined (Experiment 5) had no effect on ethanol-induced changes in neuroimmune gene expression, presumably due to poor CNS penetrance of these drugs. Finally, Experiments 6 and 7 showed that dexamethasone (subcutaneous; a GR agonist) recapitulated effects of ethanol. Overall, we conclude that ethanol-induced CORT synthesis and release is responsible for suppression of IL-1β, TNFα, and induction of IκBα in the hippocampus through GR signaling. Interventions designed to curb these changes may reduce drinking, and subdue detrimental neuroimmune activation induced by ethanol.

Human B Cells Mediate Innate Anti-Cancer Cytotoxicity Through Concurrent Engagement of Multiple TNF Superfamily Ligands.

The essential innate immunity effector cells, natural killer and dendritic cells, express multiple plasma membrane-associated tumor necrosis factor (TNF) superfamily (TNFSF) ligands that, through simultaneous and synergistic engagement, mediate anti-cancer cytotoxicity. Here, we report that circulating B cells, mediators of adaptive humoral immunity, also mediate this innate anti-cancer immune mechanism. We show that resting human B cells isolated from peripheral blood induce apoptosis of, and efficiently kill a large variety of leukemia and solid tumor cell types. Single-cell RNA sequencing analyses indicate, and flow cytometry data confirm that B cells from circulation express transmembrane TNF, Fas ligand (FasL), lymphotoxin (LT) α1β2 and TNF-related apoptosis-inducing ligand (TRAIL). The cytotoxic activity can be inhibited by individual and, especially, combined blockade of the four transmembrane TNFSF ligands. B cells from tumor-bearing head and neck squamous cell carcinoma patients express lower levels of TNFSF ligands and are less cytotoxic than those isolated from healthy individuals. In conclusion, we demonstrate that B cells have the innate capacity to mediate anti-cancer cytotoxicity through concurrent activity of multiple plasma membrane-associated TNFSF ligands, that this mechanism is deficient in cancer patients and that it may be part of a general cancer immunosurveillance mechanism.

Dusp26 phosphatase regulates mitochondrial respiration and oxidative stress and protects neuronal cell death.

The dual specificity protein phosphatases (Dusps) control dephosphorylation of mitogen-activated protein kinases (MAPKs) as well as other substrates. Here, we report that Dusp26, which is highly expressed in neuroblastoma cells and primary neurons is targetedtothe mitochondrial outermembrane via its NH2-terminal mitochondrial targeting sequence. Loss of Dusp26 has asignificant impact on mitochondrial function that is associated with increased levels of reactive oxygen species (ROS), reduction in ATP generation, reduction in mitochondria motility and release of mitochondrial HtrA2 protease into the cytoplasm. The mitochondrial dysregulation in dusp26-deficient neuroblastoma cells leads to the inhibition of cell proliferation and cell death. In vivo, Dusp26 is highly expressed in neurons in different brain regions, including cortex and midbrain (MB). Ablation of Dusp26 in mouse model leads to dopaminergic (DA) neuronal cell loss in the substantia nigra par compacta (SNpc), inflammatory response in MB and striatum, and phenotypes that are normally associated with Neurodegenerative diseases. Consistent with the data fromour mouse model, Dusp26 expressing cells are significantly reduced in the SNpc of Parkinson's Disease patients. The underlying mechanism of DA neuronal death is that loss of Dusp26 in neurons increases mitochondrial ROS and concurrent activation of MAPK/p38 signaling pathway and inflammatory response. Our results suggest that regulation of mitochondrial-associated protein phosphorylation is essential for the maintenance of mitochondrial homeostasis and dysregulation of this process may contribute to the initiation and development of neurodegenerative diseases.

The effects of immune checkpoint modulators on the clinical course of patients with resectable hepatocellular carcinoma.

Immune checkpoint proteins regulating T-cell mediated anti-tumor immunity have been reported to affect clinical outcomes in multiple malignancies. This study aimed to investigate the prognostic effect of histological expression of immune checkpoint proteins in patients with resected hepatocellular carcinoma (HCC). A total of 221 patients with HCC who underwent curative resection were included. Expression of programmed-cell death ligand-1 (PD-L1) in tumor cells (tPD-L1) and tumor infiltrating mononuclear cells (TIMCs) (iPD-L1), programmed-cell death-1 in TIMCs (iPD-1), and cytotoxic T lymphocyte antigen-4 in TIMCs (iCTLA-4) were measured immunohistochemically. Histo-positivity for iCTLA-4, iPD-1, iPD-L1, and tPD-L1 was 32.1%, 42.5%, 35.3%, and 14.9%, respectively. Multivariate logistic analyses revealed that male sex and tumor >5 cm were variables related to iCTLA-4 positivity (odds ratio [OR], 0.46 and 1.94, respectively; P<0.05). Poor differentiation was related to PD-L1 expression in both tumor cells and TIMCs (OR, 2.88 and 3.46, respectively; P<0.05). Microvascular invasion was significantly associated only with iPD-L1 (OR, 2.24; P<0.05). In time-dependent outcome analyses, expression of immune checkpoint proteins in TIMCs (i.e., iCTLA-4, iPD-1, and iPD-L1) was significantly related to longer overall survival and non-cancer-related survival (all P<0.05), but not to time-to-recurrence or cancer-specific deaths. Concurrent activation of the PD-1:PD-L1 and CTLA-4 pathways predicted improved outcomes in terms of overall survival and non-cancer related survival (P=0.06 and P=0.03, respectively). Immune checkpoint proteins upregulated in TIMCs in HCC tissues have individual and additive effects in prolonging the survival of patients, specifically in terms of survival not related to cancer recurrence.