bluuebunny/crossref_metadata_2025 huggingface dataset:
Elucidating the Mechanism of Action of CC-90009, a Novel Cereblon E3 Ligase Modulator, in AML Via Genome-Wide CRISPR Screen
Authors: Lu Gang , Surka Christine , Lu Chin-Chun , Jang In Sock , Wang Kai , Rolfe Mark
Published: February 2020

CC-90009 is a novel cereblon E3 ligase modulator (CELMoD) currently under investigation in a phase I clinical study in relapsed or refractory acute myeloid leukemia (R/R AML) (CC-90009-AML-001; NCT02848001). CC-90009 coopts the CUL4-DDB1-CRBN-RBX1 (CRL4CRBN) E3 ubiquitin ligase complex to target the translation termination factor G1 to S phase transition 1 (GSPT1) for ubiquitination and proteasomal degradation, resulting in rapid induction of apoptosis and growth inhibition in AML cell lines and primary patient blasts.

To further elucidate the mechanism of action of CC-90009 in AML, we performed a genome-wide CRISPR/Cas9 screen to identify gene(s) whose knockout abrogate(s) the response to CC-90009 in a sensitive AML cell line. In addition to well-established key regulatory proteins required for the activity of all known cereblon modulators, which include components of the CRL4CRBN complex, E2 ubiquitin conjugating enzymes UBE2G1 and UBE2D3, and members of the neddylation and deneddylation machinery, interestingly, the screen identified the ILF2 and ILF3 heterodimeric complex as a novel regulator of cereblon expression. Knockout of ILF2/ILF3 decreased the production of full-length CRBN transcript via modulating alternative splicing of CRBN mRNA, leading to significant downregulation of cereblon expression and hence diminished response to CC-90009. The screen also revealed that mTOR signaling and the integrated stress response (ISR) specifically regulate the response to CC-90009 in contrast to other cereblon modulators. Since CC-90009 inhibits protein translation, it is reasonable to expect interactions with regulators of this pathway. Hyperactivation of the mTOR pathway by inactivation of TSC1 and TSC2 protected against the growth inhibitory effect of CC-90009 , at least in part by reducing CC-90009 induced binding of GSPT1 to cereblon and subsequent GSPT1 degradation. On the other hand, GSPT1 degradation promoted the activation of the GCN1/GCN2/ATF4 pathway and subsequent apoptosis in AML cells. Loss of GCN2 significantly attenuated the growth inhibitory effect of CC-90009, and this effect can be rescued with GCN2 wild-type but not enzymatically-dead mutants.

Collectively, the antitumor activity of CC-90009, a first-in-class GSPT1 degrader, in AML cell lines is mediated by multiple layers of signaling networks and machinery, the elucidation of which reveals the underlying mechanism by which CC-90009 exerts its anti-AML activity and informs on the pathways for further study of CC-90009's clinical utility.

Disclosures

Lu: Celgene Corporation: Employment, Equity Ownership. Surka:Celgene: Employment, Equity Ownership. Lu:Celgene Corporation: Employment, Equity Ownership. Jang:Celgene: Employment, Equity Ownership. Wang:Celgene: Employment, Equity Ownership. Rolfe:Celgene: Employment, Equity Ownership.
Mutational spectrum and dynamics of clonal hematopoiesis in anemia of older individuals
Authors: van Zeventer Isabelle A. , de Graaf Aniek O. , Wouters Hanneke JCM , Van der Reijden Bert A. , van der Klauw Melanie , de Witte Theo M , Jonker Marianne A. , Malcovati Luca , Jansen Joop H , Huls Geert
Published: February 2020

Anemia is a major and currently poorly understood clinical manifestation of hematopoietic ageing. Upon ageing, hematopoietic clones harboring acquired leukemia-associated mutations expand and become detectable, now referred to as clonal hematopoiesis (CH). To investigate the relationship between CH and anemia of the elderly we explored the landscape and dynamics of CH in older anemic individuals. From the prospective population-based Lifelines cohort (n=167.729), we selected all individuals ≥60 years with anemia according to WHO criteria (n=676) and 1:1 matched controls. Peripheral blood of 1298 individuals was analyzed for acquired mutations at ≥1% variant allele frequency (VAF) in 27 driver genes. To track clonal evolution over time, we included all available follow-up samples (n=943). CH was more frequently detected in anemic individuals (46.6%) compared to controls (39.1%) (P=0.007). Although no differences were observed regarding commonly detected DTA mutations (DNMT3A, TET2, ASXL1) in anemic individuals compared to controls, other mutations were enriched in the anemia cohort, including TP53 and SF3B1. Unlike individuals with nutrient deficiency (P=0.84), individuals with anemia of chronic inflammation and unexplained anemia revealed a higher prevalence of CH (P=0.035 and P=0.017 respectively) compared to their matched controls. Follow-up analyses revealed that clones may expand and decline, generally showing only a subtle increase in VAF over 44 months (mean 0.56%), irrespective of the presence of anemia. Specific mutations associated with different growth rates and propensities to acquire an additional hit. In contrast to smaller clones (<5% VAF) which did not affect overall survival, larger clones associated with increased risk of death.
A phase 1 study of azacitidine with high-dose cytarabine and mitoxantrone in high-risk acute myeloid leukemia
Authors: Cahill Kirk E. , Karimi Yasmin H. , Karrison Theodore G. , Jain Nitin , Green Margaret , Weiner Howard , Fulton Noreen , Kadri Sabah , Godley Lucy A. , Artz Andrew S. , Liu Hongtao , Thirman Michael J. , Le Beau Michelle M. , McNerney Megan E. , Segal Jeremy , Larson Richard A. , Stock Wendy , Odenike Olatoyosi
Published: February 2020

In this phase 1 study, azacitidine (AZA) was given before high-dose cytarabine (HiDAC) and mitoxantrone (mito) based on the hypothesis that epigenetic priming with a hypomethylating agent before cytotoxic chemotherapy would improve response rates in patients with high-risk acute myeloid leukemia (AML), including relapsed/refractory disease. The primary objective was to establish the recommended phase 2 dose of AZA given before standard HiDAC/mito. In a dose escalation scheme, 46 patients (median age, 66 years) received AZA at 37.5, 50, or 75 mg/m2 subcutaneously or IV once daily on days 1 to 5 followed by HiDAC (3000 mg/m2) and mitoxantrone (30 mg/m2) once each on days 6 and 10 (the HiDAC/mito dose was reduced 33% in elderly subjects). Two dose-limiting toxicities occurred (both in the same patient): acute liver failure and kidney injury at the 50 mg/m2 dose. The 30-day induction death rate was 2.2% (1 of 46). The overall response rate, including complete remission and complete remission with incomplete count recovery, was 61% (28 of 46). Previously untreated patients aged ≥60 years with therapy-related AML and de novo AML were more likely to respond than untreated patients with AML progressing from an antecedent hematologic disorder (myelodysplastic syndrome and chronic myelomonocytic leukemia). Patients with favorable European Leukemia Network risk (P = .008), NPM1 mutations (P = .007), or IDH2 mutations (P = .03) were more likely to respond, and those with TP53 mutations (P = .03) were less likely to respond. The recommended phase 2 dose of AZA is 75 mg/m2 per day on days 1 to 5 followed by HiDAC (3000 mg/m2) and mitoxantrone (30 mg/m2) once each on days 6 and 10. This trial was registered at www.clinicaltrials.gov as #NCT01839240.
Materials selection for structured horizontal road markings: financial and environmental case studies
Authors: Burghardt Tomasz E. , Pashkevich Anton
Published: February 2020

Introduction

On majority of roads, horizontal road markings are essential safety feature that must be periodically renewed to maintain appropriate performance.

Results

The use of a premium road marking system resulted in prolonged the service life as compared to a standard system, which would be financially-neutral in the long term despite much higher unit and initial costs; advantageously, 54% less glass beads and 63% less paint would be required over 10-year life cycle. When these products usage savings are recalculated into raw materials consumption, the premium system would demand about 21% more of titanium dioxide, but that would be offset by approximately 25% reduction in the consumption of acrylic resin and circa 97% reduction in organic solvents usage and emissions.

Conclusions

The results from this analysis can be used by road administrators to select the best systems, by policy makers seeking minimisation of environmental impacts, and by contractors who won performance tenders.
Improved recovery of cell-cycle gene expression in Saccharomyces cerevisiae from regulatory interactions in multiple omics data
Authors: Panchy Nicholas L. , Lloyd John P. , Shiu Shin-Han
Published: February 2020

Background

Gene expression is regulated by DNA-binding transcription factors (TFs). Together with their target genes, these factors and their interactions collectively form a gene regulatory network (GRN), which is responsible for producing patterns of transcription, including cyclical processes such as genome replication and cell division. However, identifying how this network regulates the timing of these patterns, including important interactions and regulatory motifs, remains a challenging task.

Results

We employed four in vivo and in vitro regulatory data sets to investigate the regulatory basis of expression timing and phase-specific patterns cell-cycle expression inSaccharomyces cerevisiae. Specifically, we considered interactions based on direct binding between TF and target gene, indirect effects of TF deletion on gene expression, and computational inference. We found that the source of regulatory information significantly impacts the accuracy and completeness of recovering known cell-cycle expressed genes. The best approach involved combining TF-target and TF-TF interactions features from multiple datasets in a single model. In addition, TFs important to multiple phases of cell-cycle expression also have the greatest impact on individual phases. Important TFs regulating a cell-cycle phase also tend to form modules in the GRN, including two sub-modules composed entirely of unannotated cell-cycle regulators (STE12-TEC1andRAP1-HAP1-MSN4).

Conclusion

Our findings illustrate the importance of integrating both multiple omics data and regulatory motifs in order to understand the significance regulatory interactions involved in timing gene expression. This integrated approached allowed us to recover both known cell-cycles interactions and the overall pattern of phase-specific expression across the cell-cycle better than any single data set. Likewise, by looking at regulatory motifs in the form of TF-TF interactions, we identified sets of TFs whose co-regulation of target genes was important for cell-cycle expression, even when regulation by individual TFs was not. Overall, this demonstrates the power of integrating multiple data sets and models of interaction in order to understand the regulatory basis of established biological processes and their associated gene regulatory networks.
Presence–absence polymorphisms of single-copy genes in the stony coral Acropora digitifera
Authors: Takahashi-Kariyazono Shiho , Sakai Kazuhiko , Terai Yohey
Published: February 2020

Background

Despite the importance of characterizing genetic variation among coral individuals for understanding phenotypic variation, the correlation between coral genomic diversity and phenotypic expression is still poorly understood.

Results

In this study, we detected a high frequency of genes showing presence–absence polymorphisms (PAPs) for single-copy genes in Acropora digitifera. Among 10,455 single-copy genes, 516 (5%) exhibited PAPs, including 32 transposable element (TE)-related genes. Five hundred sixteen genes exhibited a homozygous absence in one (102) or more than one (414) individuals (n = 33), indicating that most of the absent alleles were not rare variants. Among genes showing PAPs (PAP genes), roughly half were expressed in adults and/or larvae, and the PAP status was associated with differential expression among individuals. Although 85% of PAP genes were uncharacterized or had ambiguous annotations, 70% of these genes were specifically distributed in cnidarian lineages in eumetazoa, suggesting that these genes have functional roles related to traits related to cnidarians or the family Acroporidae or the genus Acropora. Indeed, four of these genes encoded toxins that are usually components of venom in cnidarian-specific cnidocytes. At least 17% of A. digitifera PAP genes were also PAPs in A. tenuis, the basal lineage in the genus Acropora, indicating that PAPs were shared among species in Acropora.

Conclusions

Expression differences caused by a high frequency of PAP genes may be a novel genomic feature in the genus Acropora; these findings will contribute to improve our understanding of correlation between genetic and phenotypic variation in corals.
Indirect impact of violent events on emergency department utilization and disease patterns
Authors: Makhlouf-Obermeyer Carla , Sharara Eman , El-Eid Ghada , Hitti Eveline A.
Published: February 2020

Background

The health effects of war and armed conflict on casualties and mental health of those directly exposed has been well described, but few studies have explored the indirect health effects of violent events. This paper assesses the indirect health impact of several violent events that took place in Beirut in 2013–2014 on ED visit utilization and disease patterns.

Methods

As tracked by media reports, there were 9 violent events in Beirut during 2013–2014. We compared visits to the Emergency Department of a major medical center during weeks when violent events happened and weeks without such events (the preceding week and the same week in preceding years). After re-coding de-identified data from the medical records of 23,067 patients, we assessed differences in the volume of visits, severity index, and discharge diagnoses. Individual control charts were used to analyze ED visit trends post-event.

Results

Comparisons of weeks with violent events and weeks without such events indicate that the socio-demographic characteristics of patients who visited the Emergency Department were similar. Patients seen during violent weeks were significantly more likely to be admitted to the hospital, and less likely to present with low acuity complaints, indicating greater complexity of their conditions. The discharge diagnoses that were significantly higher during violent event weeks included anxiety disorders, sprains, and gastritis. Daily ED visits dropped post events by 14.111%, p < 0.0001.

Conclusions

The results indicate that violent events such as bombs, explosions, and terrorist attacks reverberate through the population, impact patterns of ED utilization immediately post-event and are associated with adverse health outcomes, even among those who are not directly affected by the events.
Appendectomy, cholecystectomy and diagnostic laparoscopy conducted before pregnancy and risk of adverse birth outcomes: a nationwide registry-based prevalence study 1996–2015
Authors: Rasmussen Anne Staub , Christiansen Christian Fynbo , Uldbjerg Niels , Nørgaard Mette
Published: February 2020

Background

Non-obstetric surgery conducted during pregnancy may increase the risk of adverse birth outcomes like small for gestational age, preterm birth, and miscarriage. Mechanisms are unclear but possibly longer lasting. We examined whether appendectomy, cholecystectomy and diagnostic laparoscopy conducted before pregnancy affect these outcomes.

Methods

This nationwide Danish prevalence study included all pregnancies during 1996–2015 that had an appendectomy, cholecystectomy or diagnostic laparoscopy registered before last menstrual period in the years 1992–2015. We excluded pregnancies with surgery and categorized pre-pregnancy surgery according to timing (0–11, 12–23, and 24+ months before last menstrual period). Outcomes were small for gestational age, late preterm birth (32–37 weeks), early preterm birth (22–31 weeks) and miscarriage (7–21 weeks). We computed absolute risks and used logistic regression comparing pregnancies with surgery 0–11 or 12–23 to 24+ months before last menstrual period, computing odds ratios for each outcome, adjusting for maternal age and smoking.

Results

We identified 15,939 pregnancies with appendectomy, 12,869 pregnancies with cholecystectomy and 19,330 pregnancies with diagnostic laparoscopy. The absolute risk of small for gestational age was 2.2% for patients with appendectomy 0–11 months before last menstrual period, 3.2% 12–23 months before compared with 2.2% when appendectomy was conducted more than 24 months before (adjusted OR 0.95 (95% CI; 0.65 to 1.31) and 1.37(95% CI;1.00 to 1.86). For early preterm birth, the absolute risks were 0.7, 0.5 and 0.8%, for late preterm birth 4.8, 4.4 and 4.7% and for miscarriage 5.7, 6.2 and 5.4%.We observed similar results for cholecystectomy. For diagnostic laparoscopy 0–11 months before pregnancy we found increased risks of small for gestational age (4.0, 2.8 and 2.6%) and late preterm birth (5.9, 5.0 and 4.8%).

Conclusions

We found no increased risk of adverse birth outcomes among pregnancies with appendectomy or cholecystectomy conducted within 2 years before pregnancy compared to more than 2 years before pregnancy. The increased risks 0–11 months after diagnostic laparoscopy are likely explained by confounding by underlying indication. It appears safe to become pregnant any time following appendectomy and cholecystectomy, but, probably depending on indication, attention should be payed 0–11 months after diagnostic laparoscopy.
Mental health symptoms and their relations with dietary diversity and nutritional status among mothers of young children in eastern Democratic Republic of the Congo
Authors: Emerson Jillian A. , Caulfield Laura E. , Kishimata Espoir Musafiri , Nzanzu Jean-Pierre , Doocy Shannon
Published: February 2020

Background

In developing countries, maternal mental health problems have been linked to sub-optimal child feeding practices and child underweight and stunting, but little is known about how maternal mental health is associated with mothers’ own diets and nutritional status. The objective of the study was to investigate the association between mental health symptoms and diet and nutritional status of mothers of young children in South Kivu, DR Congo.

Methods

Participants were 828 mothers of young children enrolled in a larger, quasi-experimental study evaluating a multi-year food security and nutrition project. The present analysis was conducted with cross-sectional data collected from 2015 to 2016. We assessed symptoms of anxiety and depression using the Hopkins Symptom Checklist-25 (HSCL-25) and post-traumatic stress disorder (PTSD) with the Harvard Trauma Questionnaire (HTQ), using a four-point Likert scale. Mean scale scores were calculated ranging from one to four. A variable was created for high distress (participants scoring in the upper quartile of both measures). Dietary diversity scores were calculated from the number of food groups (range zero to ten) consumed the previous day, identified from an open recall. Nutritional status was measured by body mass index (BMI) and underweight (BMI < 18.5 kg/m2, or mid-upper arm circumference < 23 cm for pregnant women). Bivariate and multivariate (adjusting for parent study intervention group, education, age, health, parity, livelihoods zone, and territory of origin) regression analyses were conducted.

Results

Maternal mental health measures were positively and statistically significantly associated with higher dietary diversity scores in adjusted analyses (HSCL-25: ß= 0.18, p = 0.002, HTQ: ß= 0.12, p = 0.029, High Distress: ß= 0.47, p < 0.001). Mental health symptoms were not significantly associated with BMI (HSCL-25: ß = − 0.04, p = 0.824; HTQ: ß = 0.02, p = 0.913; High distress: ß= − 0.02, p = 0.938) or underweight (HSCL 25: OR = 0.91, p = 0.640; HTQ: OR = 1.03, p = 0.866; High distress: OR = 0.78, p = 0.489).

Conclusions

More severe maternal mental health symptoms were associated with higher dietary diversity but not nutritional status, and the reasons for these findings are not clear from available data. More research is needed to identify underlying factors that could influence mental health symptomatology and diet quality among food insecure and extremely resource-limited populations.
Low skeletal muscle mass is associated with arterial stiffness in community-dwelling Chinese aged 45 years and older
Authors: Yang Mingzhe , Zhang Xuguang , Ding Zhenkai , Wang Feijie , Wang Yufang , Jiao Changya , Chen Jie-Hua
Published: February 2020

Background

Evidence suggests that body composition has impact on arterial stiffness. However, evidence in Chinese are limited, and results remain controversial. The aim of our study is to investigate whether skeletal muscle mass is associated with arterial stiffness in Chinese community-dwelling men and women aged 45 years and older.

Methods

In this cross-sectional study, 20,477 participants (age range: 45–80 years, 68.8% women) were included in the analysis. Brachial-ankle pulse wave velocity (baPWV), an indicator of arterial stiffness was measured using a waveform device. Total muscle mass and muscle mass of arm, leg and trunk were measured by bioelectrical impedance analysis. Height and weight were measured and appendicular skeletal muscle mass index (ASMI) was calculated as appendicular skeletal muscle mass (sum of arm and leg muscle mass) divided by height square.

Results

After adjustment for age, body fat percentage, systolic blood pressure and diastolic blood pressure, ASMI was negatively associated with baPWV [β (SE) for men: − 0.208 (0.016),p < 0.0001; for women: − 0.245 (0.012),p < 0.0001]. High ASMI was a protective factor for the presence of arterial stiffness (defined as baPWV) [OR (95%CI) for men: 0.730 (0.682, 0.782),p < 0.0001; women: 0.634 (0.593, 0.677),p < 0.0001]. Similar associations were found between quantity of muscle mass (total and appendicular muscle mass, muscle mass of arm, leg and trunk) and arterial stiffness in men and women after further adjustment for height (allp < 0.0001).

Conclusion

Low skeletal muscle mass is associated with increased risk of arterial stiffness in Chinese community-dwelling adults aged 45 years and older.
The complex neurobiology of resilient functioning after childhood maltreatment
Authors: Ioannidis Konstantinos , Askelund Adrian Dahl , Kievit Rogier A. , van Harmelen Anne-Laura
Published: February 2020

Background

Childhood maltreatment has been associated with significant impairment in social, emotional and behavioural functioning later in life. Nevertheless, some individuals who have experienced childhood maltreatment function better than expected given their circumstances.

Main body

Here, we provide an integrated understanding of the complex, interrelated mechanisms that facilitate such individual resilient functioning after childhood maltreatment. We aim to show that resilient functioning is not facilitated by any single ‘resilience biomarker’. Rather, resilient functioning after childhood maltreatment is a product of complex processes and influences across multiple levels, ranging from ‘bottom-up’ polygenetic influences, to ‘top-down’ supportive social influences. We highlight the complex nature of resilient functioning and suggest how future studies could embrace a complexity theory approach and investigate multiple levels of biological organisation and their temporal dynamics in a longitudinal or prospective manner. This would involve using methods and tools that allow the characterisation of resilient functioning trajectories, attractor states and multidimensional/multilevel assessments of functioning. Such an approach necessitates large, longitudinal studies on the neurobiological mechanisms of resilient functioning after childhood maltreatment that cut across and integrate multiple levels of explanation (i.e. genetics, endocrine and immune systems, brain structure and function, cognition and environmental factors) and their temporal interconnections.

Conclusion

We conclude that a turn towards complexity is likely to foster collaboration and integration across fields. It is a promising avenue which may guide future studies aimed to promote resilience in those who have experienced childhood maltreatment.
Incremental value of three-dimensional echocardiography for evaluating left atrial function in patients with coronary slow flow phenomenon: a case control study
Authors: Li Jun , Wang Yonghuai , Zhao Cuiting , Zhu Qing , Li Guangyuan , Yang Jun , Jia Dalin , Ma Chunyan
Published: February 2020

Background

Coronary slow flow phenomenon (CSFP) involves the delayed opacification of the coronary distal vessel, in the absence of an obstructive lesion in the epicardial coronary artery during angiography. Since the link between left atrial (LA) function and decreased left ventricular function is still unclear, we evaluated LA function using real-time three-dimensional echocardiography (RT3DE) in patients with CSFP, and subsequently determined the incremental value of RT3DE.

Methods

This study enrolled 60 patients with CSFP and 45 control subjects. CSFP was diagnosed based on thrombolysis in myocardial infarction frame count (TFC). The LA phasic volume and function was evaluated by both two-dimensional echocardiography (2DE) and RT3DE.

Results

The LA maximal volume (Volmax), pre-systolic volume (Volp), and minimal volume (Volmin) increased, but LA total and active ejection fraction decreased in patients with CSFP. Based on our results, Volmax, Volp, Volmin, and LA total and active ejection fraction correlated with TFC, and with the number of arteries involved. The LA total ejection fraction by RT3DE was the only independent predictor for CSFP (odds ratio, 0.64 [95% confidence interval, 0.49–0.83]; P = 0.001). Also, the LA total ejection fraction by RT3DE demonstrated good predictive power for CSFP, with a cut-off value of 54.15% (area under curve, 0.85; sensitivity, 84%; specificity, 83%).

Conclusions

The LA reservoir and contractile function decreased in the patients with CSFP and correlated with coronary flow rate and with the number of arteries involved. The LA total ejection fraction by RT3DE can independently predict CSFP, and RT3DE demonstrated incremental value for evaluating LA phasic function in the patients with CSFP compared to 2DE.
Low-intensity ultrasound attenuates paw edema formation and decreases vascular permeability induced by carrageenan injection in rats
Authors: Kim Kil Hwan , Im Hyeon-Woo , Karmacharya Mrigendra Bir , Kim Sejong , Min Byoung-Hyun , Park So Ra , Choi Byung Hyune
Published: February 2020

Background

Therapeutic potential of low-intensity ultrasound (LIUS) has become evident in various musculoskeletal diseases. We have previously shown that LIUS has an inhibitory effect on local edema in various diseases including the arthritis and brain injury. In this study, we examined whether LIUS can attenuate paw edema formation vis-à-vis vascular permeability and inflammation in rats induced by carrageenan. LIUS with a frequency of 1 MHz and the intensities of 50, 100, or 200 mW/cm2 were exposed on rat paws for 10 min immediately after carrageenan injection.

Results

Carrageenan injection induced paw edema which was peaked at 6 h and gradually decreased nearly to the initial baseline value after 72 h. LIUS showed a significant reduction of paw edema formation at 2 and 6 h at all intensities tested. The highest reduction was observed at the intensity of 50 mW/cm2. Histological analyses confirmed that LIUS clearly decreased the carrageenan-induced swelling of interstitial space under the paw skin and infiltration of polymorphonuclear leukocytes. Moreover, Evans Blue extravasation analyses exhibited a significant decreases of vascular permeability by LIUS. Finally, immunohistochemical staining showed that expression of pro-inflammatory proteins, namely, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) induced by carrageenan injection was reduced back to the normal level after LIUS stimulation.

Conclusions

These results provide a new supporting evidence for LIUS as a therapeutic alternative for the treatment of edema in inflammatory diseases such as cellulitis.
The eEF2 kinase-induced STAT3 inactivation inhibits lung cancer cell proliferation by phosphorylation of PKM2
Authors: Xiao Min , Xie Jianling , Wu Yu , Wang Genzhu , Qi Xin , Liu Zailiang , Wang Yuying , Wang Xuemin , Hoque Ashfaqul , Oakhill Jon , Proud Christopher G. , Li Jing
Published: February 2020

Background

Eukaryotic elongation factor-2 kinase (eEF2K) is a Ca 2+ /calmodulin (CaM)-dependent protein kinase that inhibits protein synthesis. However, the role of eEF2K in cancer development was reported paradoxically and remains to be elucidated.

Methods

Herein, A549 cells with eEF2K depletion or overexpression by stably transfected lentivirus plasmids were used in vitro and in vivo study. MTT and colony assays were used to detect cell proliferation and growth. Extracellular glucose and lactate concentration were measured using test kit. Immunoblot and co-immunoprecipitation assays were used to examine the molecular biology changes and molecular interaction in these cells. LC-MS/MS analysis and [γ- 32 P] ATP kinase assay were used to identify combining protein and phosphorylation site. Nude mice was utilized to study the correlation of eEF2K and tumor growth in vivo.

Results

We demonstrated that eEF2K inhibited lung cancer cells proliferation and affected the inhibitory effects of EGFR inhibitor gefitinib. Mechanistically, we showed that eEF2K formed a complex with PKM2 and STAT3, thereby phosphorylated PKM2 at T129, leading to reduced dimerization of PKM2. Subsequently, PKM2 impeded STAT3 phosphorylation and STAT3-dependent c-Myc expression. eEF2K depletion promoted the nuclear translocation of PKM2 and increased aerobic glycolysis reflected by increased lactate secretion and glucose.

Conclusions

Our findings define a novel mechanism underlying the regulation of cancer cell proliferation by eEF2K independent of its role in protein synthesis, disclosing the diverse roles of eEF2K in cell biology, which lays foundation for the development of new anticancer therapeutic strategies.
Clinical, pathological and dermoscopic phenotype of MITF p.E318K carrier cutaneous melanoma patients
Authors:   Italian Melanoma Intergroup (I.M.I.), Ciccarese Giulia , Dalmasso Bruna , Bruno William , Queirolo Paola , Pastorino Lorenza , Andreotti Virginia , Spagnolo Francesco , Tanda Enrica , Ponti Giovanni , Massone Cesare , Drago Francesco , Parodi Aurora , Ghigliotti Giovanni , Pizzichetta Maria Antonietta , Ghiorzo Paola
Published: February 2020

Background

The p.E318K variant of the Melanocyte Inducing Transcription Factor (MITF) has been implicated in genetic predisposition to melanoma as an intermediate penetrance allele. However, the impact of this variant on clinico-phenotypic, as well as on dermoscopic patterns features of affected patients is not entirely defined. The purpose of our study was to assess the association between the p.E318K germline variant and clinic-phenotypical features of MITF+ compared to non-carriers (MITF−), including dermoscopic findings of melanomas and dysplastic nevi.

Methods

we retrospectively analyzed a consecutive series of 1386 patients recruited between 2000 and 2017 who underwent genetic testing for CDKN2A, CDK4, MC1R and MITF germline variants in our laboratory for diagnostic/research purposes. The patients were probands of melanoma-prone families and apparently sporadic single or multiple primary melanoma patients. For all, we collected clinical, pathological information and dermoscopic images of the histopathologically diagnosed melanomas and dysplastic nevi, when available.

Results

After excluding patients positive for CDKN2A/CDK4 pathogenic variants and those affected by non-cutaneous melanomas, our study cohort comprised 984 cutaneous melanoma patients, 22 MITF+ and 962 MITF−. MITF+ were more likely to develop dysplastic nevi and multiple primary melanomas. Nodular melanoma was more common in MITF+ patients (32% compared to 19% in MITF−). MITF+ patients showed more frequently dysplastic nevi and melanomas with uncommon dermoscopic patterns (unspecific), as opposed to MITF− patients, whose most prevalent pattern was the multicomponent.

Conclusions

MITF+ patients tend to develop melanomas and dysplastic nevi with histopathological features, frequency and dermoscopic patterns often different from those prevalent in MITF− patients. Our results emphasize the importance of melanoma prevention programs for MITF+ patients, including dermatologic surveillance with digital follow-up.
YAP1 is an independent prognostic marker in pancreatic cancer and associated with extracellular matrix remodeling
Authors: Zhou Qimin , Bauden Monika , Andersson Roland , Hu Dingyuan , Marko-Varga György , Xu Jianfeng , Sasor Agata , Dai Hua , Pawłowski Krzysztof , Said Hilmersson Katarzyna , Chen Xi , Ansari Daniel
Published: February 2020

Background

Pancreatic cancer is a major cause of cancer-related mortality. The identification of effective biomarkers is essential in order to improve management of the disease. Yes-associated protein 1 (YAP1) is a downstream effector of the Hippo pathway, a signal transduction system implicated in tissue repair and regeneration, as well as tumorigenesis. Here we evaluate the biomarker potential of YAP1 in pancreatic cancer tissue.

Methods

YAP1 was selected as a possible biomarker for pancreatic cancer from global protein sequencing of fresh frozen pancreatic cancer tissue samples and normal pancreas controls. The prognostic utility of YAP1 was evaluated using mRNA expression data from 176 pancreatic cancer patients in The Cancer Genome Atlas (TCGA), as well as protein expression data from immunohistochemistry analysis of a local tissue microarray (TMA) cohort comprising 140 pancreatic cancer patients. Ingenuity Pathway Analysis was applied to outline the interaction network for YAP1 in connection to the pancreatic tumor microenvironment. The expression of YAP1 target gene products was evaluated after treatment of the pancreatic cancer cell line Panc-1 with three substances interrupting YAP–TEAD interaction, including Super-TDU, Verteporfin and CA3.

Results

Mass spectrometry based proteomics showed that YAP1 is the top upregulated protein in pancreatic cancer tissue when compared to normal controls (log2 fold change 6.4; p = 5E−06). Prognostic analysis of YAP1 demonstrated a significant correlation between mRNA expression level data and reduced overall survival (p = 0.001). In addition, TMA and immunohistochemistry analysis suggested that YAP1 protein expression is an independent predictor of poor overall survival [hazard ratio (HR) 1.870, 95% confidence interval (CI) 1.224–2.855, p = 0.004], as well as reduced disease-free survival (HR 1.950, 95% CI 1.299–2.927, p = 0.001). Bioinformatic analyses coupled with in vitro assays indicated that YAP1 is involved in the transcriptional control of target genes, associated with extracellular matrix remodeling, which could be modified by selected substances disrupting the YAP1-TEAD interaction.

Conclusions

Our findings indicate that YAP1 is an important prognostic biomarker for pancreatic cancer and may play a regulatory role in the remodeling of the extracellular matrix.
A very low thymus function identifies patients with substantial increased risk for long-term mortality after kidney transplantation
Authors: Betjes Michiel G. H. , Langerak Anton W. , Klepper Mariska , Litjens Nicolle H. R.
Published: February 2020

Background

End-stage renal disease is associated with premature ageing of the T cell immune system but inter-individual variation is substantial. The hypothesis was tested that advanced immunological T cell ageing assessed by peripheral T cell differentiation increases the long-term mortality risk after renal transplantation.

Results

Circulating T cells of 211 recipients of a kidney from a living donor were analyzed before and in the first year after transplantation. The number of CD31-positive naive T cells (as a marker for recent thymic emigrants) and the differentiation status of the memory T cells was assessed. Thirty recipients died during follow-up of at least 5 years. Absolute numbers of naive CD4+(living:258 cells/μl vs. deceased:101 cells/μl,p < 0.001) and naive CD8+T cells (living:97 cells/μl vs. deceased:37 cells/μl, p < 0.001) were significantly lower in the deceased group prior to transplantation. In a multivariate proportional hazard analysis the number of naive CD4+T cells remained associated with all-cause mortality (HR 0.98, CI 0.98–0.99,p < 0.001). The low number of naive T cells in the deceased patient group was primarily caused by a decrease in recent thymic emigrants (i.e. less CD31+naive T cells) indicating a lowered thymus function. In addition, the physiological age-related compensatory increase in CD31−naïve T cells was not observed. Within the first year after transplantation, the number and characteristics of naive T cells remained stable.

Conclusions

A severe reduction in circulating naïve T cells because of a decrease in recent thymic emigrants is highly associated with all-cause mortality after renal transplantation.
Assessment of bilateral motor skills and visuospatial attention in children with perinatal stroke using a robotic object hitting task
Authors: Hawe Rachel L. , Kuczynski Andrea M. , Kirton Adam , Dukelow Sean P.
Published: February 2020

Background

While motor deficits are the hallmark of hemiparetic cerebral palsy, children may also experience impairments in visuospatial attention that interfere with participation in complex activities, including sports or driving. In this study, we used a robotic object hitting task to assess bilateral sensorimotor control and visuospatial skills in children with hemiparesis due to perinatal arterial ischemic stroke (AIS) or periventricular venous infarct (PVI). We hypothesized that performance would be impaired bilaterally and be related to motor behavior and clinical assessment of visuospatial attention.

Methods

Forty-nine children with perinatal stroke and hemiparetic cerebral palsy and 155 typically developing (TD) children participated in the study. Participants performed a bilateral object hitting task using the KINARM Exoskeleton Robot, in which they used virtual paddles at their fingertips to hit balls that fell from the top of the screen with increasing speed and frequency over 2.3 min. We quantified performance across 13 parameters including number of balls hit with each hand, movement speed and area, biases between hands, and spatial biases. We determined normative ranges of performance accounting for age by fitting 95% prediction bands to the TD children. We compared parameters between TD, AIS, and PVI groups using ANCOVAs accounting for age effects. Lastly, we performed regression analysis between robotic and clinical measures.

Results

The majority of children with perinatal stroke hit fewer balls with their affected arm compared to their typically developing peers. We also found deficits with the ipsilesional (“unaffected”) arm. Children with AIS had greater impairments than PVI. Despite hitting fewer balls, we only identified 18% of children as impaired in hand speed or movement area. Performance on the Behavioral Inattention Test accounted for 21–32% of the variance in number of balls hit with the unaffected hand.

Conclusions

Children with perinatal stroke-induced hemiparetic cerebral palsy may have complex bilateral deficits reflecting a combination of impairments in motor skill and visuospatial attention. Clinical assessments and interventions should address the interplay between motor and visuospatial skills.
Drought and heat stress tolerance screening in wheat using computed tomography
Authors: Schmidt Jessica , Claussen Joelle , Wörlein Norbert , Eggert Anja , Fleury Delphine , Garnett Trevor , Gerth Stefan
Published: February 2020

Background

Improving abiotic stress tolerance in wheat requires large scale screening of yield components such as seed weight, seed number and single seed weight, all of which is very laborious, and a detailed analysis of seed morphology is time-consuming and visually often impossible. Computed tomography offers the opportunity for much faster and more accurate assessment of yield components.

Results

An X-ray computed tomographic analysis was carried out on 203 very diverse wheat accessions which have been exposed to either drought or combined drought and heat stress. Results demonstrated that our computed tomography pipeline was capable of evaluating grain set with an accuracy of 95–99%. Most accessions exposed to combined drought and heat stress developed smaller, shrivelled seeds with an increased seed surface. As expected, seed weight and seed number per ear as well as single seed size were significantly reduced under combined drought and heat compared to drought alone. Seed weight along the ear was significantly reduced at the top and bottom of the wheat spike.

Conclusions

We were able to establish a pipeline with a higher throughput with scanning times of 7 min per ear and accuracy than previous pipelines predicting a set of agronomical important seed traits and to visualize even more complex traits such as seed deformations. The pipeline presented here could be scaled up to use for high throughput, high resolution phenotyping of tens of thousands of heads, greatly accelerating breeding efforts to improve abiotic stress tolerance.
Evaluation of plan quality and robustness of IMPT and helical IMRT for cervical cancer
Authors: Shang Haijiao , Pu Yuehu , Wang Wei , Dai Zhitao , Jin Fu
Published: February 2020

Background

Both plan quality and robustness were investigated through comparing some dosimetric metrics between intensity modulated proton therapy (IMPT) and helical tomotherapy based intensity modulated radiotherapy (IMRT) for cervical cancer.

Methods

Both a spot-scanning robust (SRO) IMPT plan and a helical tomotherapy robust (TRO) IMRT plan were generated for each of 18 patients. In order to evaluate the quality of nominal plans without dose perturbations, planning scores (PS) on clinical target volume (CTV) and five organs at risk (OARs) based on clinical experience, and normal tissue complication probabilities (NTCP) of rectum and sigmoid were calculated based on Lyman-Kutcher-Burman (LKB) model. Dose volume histogram bands width (DVHBW) were calculated in 28 perturbed scenarios to evaluate plan robustness.

Results

Compared with TRO, the average scores of SRO nominal plans were higher in target metrics [V46.8Gy, V50Gy, Conformity and Homogeneity](16.5 vs. 15.1), and in OARs metrics (60.9 vs. 53.3), including bladder [V35,V45, Dmean,D2cc], rectum [V40,V45,D2cc,Dmax], bowel [V35,V40,V45, Dmax], sigmoid [V40,Dmax] and femoral heads [V30,Dmax]. Meanwhile, NTCP calculation showed that the toxicities of rectum and sigmoid in SRO were lower than those in TRO (rectum: 2.8% vs. 4.8%, p < 0.05; sigmoid: 5.2% vs. 5.7%, p < 0.05). DVHBW in target coverage for the SRO plan was smaller than that for the TRO plan (0.6% vs. 2.1%), which means that the SRO plan generated a more robust plan in target.

Conclusion

Better CTV coverage and OAR Sparing were obtained in SRO nominal plan. Based on NTCP calculation, SRO was expected to allow a small reduction in rectal toxicity. Furthermore, SRO generated a more robust plan in CTV target coverage.
Validation of the graded prognostic assessment for gastrointestinal cancers with brain metastases (GI-GPA)
Authors: Nieder Carsten , Hintz Mandy , Popp Ilinca , Bilger Angelika , Grosu Anca L.
Published: February 2020

Purpose

The purpose of this study was to validate a new prognostic model (GI-GPA) originally derived from a multi-center database (USA, Canada, Japan).

Patients and Methods

This retrospective study included 92 German and Norwegian patients treated with individualized approaches, always including brain radiotherapy. Information about age, extracranial spread, number of brain metastases, performance status and other variables was collected. The GI-GPA score was calculated as described by Sperduto et al.

Results

Median survival was 4 months. The corresponding figures for the 4 different prognostic strata were 2.3, 4.4, 9.4 and 12.7 months, respectively (p = 0.0001). Patients whose management included surgical resection had longer median survival than those who were treated with other approaches (median 11.9 versus 3.0 months, p = 0.002). Comparable results were seen for additional systemic therapy (median 8.5 versus 3.5 months, p = 0.01).

Conclusion

These results confirm the validity of the GI-GPA in an independent dataset from a different geographical region, despite the fact that overall survival was shorter in all prognostic strata, compared to Sperduto et al. Potential explanations include differences in molecular tumor characteristics and treatment selection, both brain metastases-directed and extracranially. Long-term survival beyond 5 years is possible in a small minority of patients.
Umbilical cord-derived Wharton’s jelly for regenerative medicine applications
Authors: Gupta Ashim , El-Amin Saadiq F. , Levy Howard J. , Sze-Tu Rebecca , Ibim Sobrasua E. , Maffulli Nicola
Published: February 2020

Background

The last decade has seen an explosion in the interest in using biologics for regenerative medicine applications, including umbilical cord-derived Wharton’s Jelly. There is insufficient literature assessing the amount of growth factors, cytokines, hyaluronic acid, and extracellular vesicles including exosomes in these products. The present study reports the development of a novel Wharton’s jelly formulation and evaluates the presence of growth factors, cytokines, hyaluronic acid, and extracellular vesicles including exosomes.

Methods

Human umbilical cords were obtained from consenting caesarian section donors. The Wharton’s jelly was then isolated from the procured umbilical cord and formulated into an injectable form. Randomly selected samples from different batches were analyzed for sterility testing and to quantify the presence of growth factors, cytokines, hyaluronic acid, and extracellular vesicles.

Results

All samples passed the sterility test. Growth factors including IGFBP 1, 2, 3, 4, and 6, TGF-α, and PDGF-AA were detected. Several immunomodulatory cytokines, such as RANTES, IL-6R, and IL-16, were also detected. Pro-inflammatory cytokines MCSFR, MIP-1a; anti-inflammatory cytokines TNF-RI, TNF-RII, and IL-1RA; and homeostatic cytokines TIMP-1 and TIMP-2 were observed. Cytokines associated with wound healing, ICAM-1, G-CSF, GDF-15, and regenerative properties, GH, were also expressed. High concentrations of hyaluronic acid were observed. Particles in the extracellular vesicle size range were also detected and were enclosed by the membrane, indicative of true extracellular vesicles.

Conclusion

There are numerous growth factors, cytokines, hyaluronic acid, and extracellular vesicles present in the Wharton’s jelly formulation analyzed. The amount of these factors in Wharton’s jelly is higher compared with other biologics and may play a role in reducing inflammation and pain and augment healing of musculoskeletal injuries.