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Abstract A combination of improved body armor, medical transportation, and treatment has led to the increased survival of warfighters from combat extremity injuries predominantly caused by blasts in modern conflicts. Despite advances, a high rate of complications such as wound infections, wound failure, amputations, and a decreased quality of life exist. To study the molecular underpinnings of wound failure, wound tissue biopsies from combat extremity injuries had RNA extracted and sequenced. Wounds were classified by colonization (colonized vs. non-colonized) and outcome (healed vs. failed) status. Differences in gene expression were investigated between timepoints at a gene level, and longitudinally by multi-gene networks, inferred proportions of immune cells, and expression of healing-related functions. Differences between wound outcomes in colonized wounds were more apparent than in non-colonized wounds. Colonized/healed wounds appeared able to mount an adaptive immune response to infection and progress beyond the inflammatory stage of healing, while colonized/failed wounds did not. Although, both colonized and non-colonized failed wounds showed increasing inferred immune and inflammatory programs, non-colonized/failed wounds progressed beyond the inflammatory stage, suggesting different mechanisms of failure dependent on colonization status. Overall, these data reveal gene expression profile differences in healing wounds that may be utilized to improve clinical treatment paradigms.

The spraying method offers the advantages of environmental protection, convenient construction, and cost-effectiveness, making it widely used in pipeline structure repair. Given the complex service environment of pipelines, there is an urgent demand for high-strength and durable spraying materials. To achieve this goal, a PVA fiber reinforced cement mortar material (PFCM) with high early strength, excellent impermeability and stable acid corrosion resistance was developed through orthogonal test design. The optimal material was obtained through factors such as setting time, fluidity, compressive strength and flexural strength. The effects of the contents of fly ash, redispersible polymer powder and PVA fiber on the working and mechanical properties of the material were investigated, and further research was conducted on its tensile strength, impermeability, acid corrosion resistance. In addition, Thermogravimetric-differential Thermal Analysis (TG-DTA) was utilized to analyze the thermal stability of PFCM. X-ray diffraction (XRD) was utilized to analyze the composition of PFCM. Mercury intrusion porosimetry (MIP) was utilized to analyze the pore structure of PFCM, and the feasibility of the PFCM was verified. The results indicate that the contents of fly ash, redispersible polymer powder and PVA fiber have significant effects on the working and mechanical properties. The optimum initial setting time is 60–80 min, and the optimum fluidity is between 170 and 180 mm. The compressive strength and flexural strength of the mortar prepared with the optimum parameters are 102.5 MPa and 20 MPa, respectively. The material has an excellent spraying effect with no sagging, and exhibits high early strength, outstanding corrosion resistance, low porosity, and exceptional impermeability. This study lays the foundation for achieving high-quality spray repairs of underground drainage pipelines.

Summary: Human genetic studies of critical COVID-19 pneumonia have revealed the essential role of type I interferon-dependent innate immunity to SARS-CoV-2 infection. Conversely, an association between the HLA-B∗15:01 allele and asymptomatic SARS-CoV-2 infection in unvaccinated individuals was recently reported, suggesting a contribution of pre-existing T cell-dependent adaptive immunity. We report a lack of association of classical HLA alleles, including HLA-B∗15:01, with pre-omicron asymptomatic SARS-CoV-2 infection in unvaccinated participants in a prospective population-based study in the United States (191 asymptomatic vs. 945 symptomatic COVID-19 cases). Moreover, we found no such association in the international COVID Human Genetic Effort cohort (206 asymptomatic vs. 574 mild or moderate COVID-19 cases and 1,625 severe or critical COVID-19 cases). Finally, in the Human Challenge Characterisation study, the three HLA-B∗15:01 individuals infected with SARS-CoV-2 developed symptoms. As with other acute primary infections studied, no classical HLA alleles favoring an asymptomatic course of SARS-CoV-2 infection were identified.

Hard petroleum asphalt (HPA) has good application prospects because of its simple manufacturing process and excellent performance. However, according to previous studies, the performance of HPA at low temperatures cannot be well characterised by commonly used methods. The use of HPA in cold climates is severely constrained. In order to comprehensively evaluate the low-temperature performance of the material, this study used four different types of HPA as the research object, and used the bending beam rheological test ( BBR) to determine the basic rheological properties of the material. On this basis, after fitting and analysing the low-temperature creep compliance curve using the Burgers viscoelastic model, the asphalt was assigned a low-temperature viscoelastic assessment index. The glass transition temperature and sensitivity of HPA were investigated by differential scanning calorimetry (DSC). At the same time, the HPA assessment index at low temperatures was determined systematically by using grey correlation theory.

BackgroundNormal hepatic functional reserve is the key to avoiding liver failure after liver surgery. This study investigated the assessment of hepatic functional reserve using liver shear wave velocity (LSWV) combined with biochemical indicators, tumor volume, and portal vein diameter.MethodsIn this single-center prospective study, a total of 123 patients with hepatocellular carcinoma (HCC) were divided into a test group (n=92) and a validation group (n=31). All patients were Child-Pugh grade A. The indocyanine green retention rate at 15 min (ICG-R15), liver shear wave velocity (LSWV), portal vein diameter (Dpv), alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (γ-GGT), albumin (ALB), prothrombin time (PT), and also liver tumor volume (maximum diameter ≤5 cm) were measured. In the test group, multiple parameters were used to evaluate hepatic functional reserve, and the multiparametric model was established. Receiver operating characteristic (ROC) curve analysis was conducted to assess the diagnostic performance of the multiparametric model. In the validation group, the predictive effectiveness of the multiparametric model was analyzed using consistency tests.ResultsIt was revealed that LSWV, ALB, and PT were statistically significant in evaluation of the hepatic functional reserve (P

Abstract Introduction Parameters, such as left ventricular ejection fraction, peak strain dispersion, global longitudinal strain, etc. are influential and clinically interpretable for detection of cardiac disease, while manual detection requires laborious steps and expertise. In this study, we evaluated a video-based deep learning method that merely depends on echocardiographic videos from four apical chamber views of hypertensive cardiomyopathy detection. Methods One hundred eighty-five hypertensive cardiomyopathy (HTCM) patients and 112 healthy normal controls (N) were enrolled in this diagnostic study. We collected 297 de-identified subjects’ echo videos for training and testing of an end-to-end video-based pipeline of snippet proposal, snippet feature extraction by a three-dimensional (3-D) convolutional neural network (CNN), a weakly-supervised temporally correlated feature ensemble, and a final classification module. The snippet proposal step requires a preliminarily trained end-systole and end-diastole timing detection model to produce snippets that begin at end-diastole, and involve contraction and dilatation for a complete cardiac cycle. A domain adversarial neural network was introduced to systematically address the appearance variability of echo videos in terms of noise, blur, transducer depth, contrast, etc. to improve the generalization of deep learning algorithms. In contrast to previous image-based cardiac disease detection architectures, video-based approaches integrate spatial and temporal information better with a more powerful 3D convolutional operator. Results Our proposed model achieved accuracy (ACC) of 92%, area under receiver operating characteristic (ROC) curve (AUC) of 0.90, sensitivity(SEN) of 97%, and specificity (SPE) of 84% with respect to subjects for hypertensive cardiomyopathy detection in the test data set, and outperformed the corresponding 3D CNN (vanilla I3D: ACC (0.90), AUC (0.89), SEN (0.94), and SPE (0.84)). On the whole, the video-based methods remarkably appeared superior to the image-based methods, while few evaluation metrics of image-based methods exhibited to be more compelling (sensitivity of 93% and negative predictive value of 100% for the image-based methods (ES/ED and random)). Conclusion The results supported the possibility of using end-to-end video-based deep learning method for the automated diagnosis of hypertensive cardiomyopathy in the field of echocardiography to augment and assist clinicians. Trial registration Current Controlled Trials ChiCTR1900025325, Aug, 24, 2019. Retrospectively registered.

The aim of this study was to explore alterations in plasma metabolites among mares afflicted with endometritis. Mares were divided into two groups, namely, the equine endometritis group (n = 8) and the healthy control group (n = 8), which included four pregnant and four non-pregnant mares, using a combination of clinical assessment and laboratory confirmation. Plasma samples from both groups of mares were analyzed through untargeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics. A total of 28 differentially abundant metabolites were identified by screening and identifying differentially abundant metabolites and analyzing the pathway enrichment of differentially. Ten metabolites were identified as potential biomarkers for the diagnosis of endometritis in mares. Among them, seven exhibited a decrease in the endometritis groups, including hexadecanedioic acid, oleoyl ethanolamide (OEA), [fahydroxy(18:0)]12_13-dihydroxy-9z-octa (12,13-diHOME), deoxycholic acid 3-glucuronide (DCA-3G), 2-oxindole, and (+/-)9-HPODE, and 13(S)-HOTRE. On the other hand, three metabolites, adenosine 5′-monophosphate (AMP), 5-hydroxy-dl-tryptophan (5-HTP), and l-formylkynurenine, demonstrated an increase. These substances primarily participate in the metabolism of tryptophan and linolenic acid, as well as fat and energy. In conclusion, metabolomics revealed differentially abundant metabolite changes in patients with mare endometritis. These specific metabolites can be used as potential biomarkers for the non-invasive diagnosis of mare endometritis.

Zearalenone (ZEA) is a common non-steroidal estrogenic mycotoxin found in a range of animal feeds and poses a serious threat to the reproductive health of farm animals and humans. However, the mechanism underlying ZEA-induced reproductive toxicity in sheep remains unknown. Granulosa cells are crucial for egg maturation and the fertility of female sheep. In this study, we aimed to examine the impact of different ZEA concentrations on sheep follicular granulosa cells and to elucidate the potential molecular mechanism underlying ZEA-induced toxicity using transcriptome sequencing and molecular biological approaches. Treating primary sheep follicular granulosa cells with different concentrations of ZEA promoted the overproduction of reactive oxygen species (ROS), increased lipid peroxidation products, led to cellular oxidative stress, decreased antioxidant enzyme activities, and induced cell apoptosis. Using transcriptome approaches, 1395 differentially expressed genes were obtained from sheep follicular granulosa cells cultured in vitro after ZEA treatment. Among them, heme oxygenase-1 (HMOX1) was involved in 11 biological processes. The protein interaction network indicated interactions between HMOX1 and oxidative and apoptotic proteins. In addition, N-acetylcysteine pretreatment effectively reduced the ZEA-induced increase in the expression of HMOX1 and Caspase3 by eliminating ROS. Hence, we suggest that HMOX1 is a key differential gene involved in the regulation of ZEA-induced oxidative stress and apoptosis in follicular granulosa cells. These findings provide novel insights into the prevention and control of mycotoxins in livestock.

BackgroundLiver cirrhosis is closely associated with cardiac dysfunction. The aims of this study were to evaluate left ventricular systolic function in patients with hepatitis B cirrhosis by non-invasive left ventricular pressure-strain loop (LVPSL) technique, and to explore the correlation between myocardial work indices and liver function classification.MethodsAccording to the Child-Pugh classification, 90 patients with hepatitis B cirrhosis were further divided into three groups: Child-Pugh A group (n = 32), Child-Pugh B group (n = 31), and Child-Pugh C group (n = 27). During the same period, 30 healthy volunteers were recruited as the control (CON) group. Myocardial work parameters, which included global work index (GWI), global constructive work (GCW), global wasted work (GWW), and global work efficiency (GWE), were derived from the LVPSL and compared among the four groups. The correlation between myocardial work parameters and Child-Pugh liver function classification was evaluated, and the independent risk factors affecting left ventricular myocardial work in patients with cirrhosis were investigated by univariable and multivariable linear regression analysis.ResultsGWI, GCW and GWE of Child-Pugh B and C groups were lower than those of CON group, while GWW was higher than that of CON group, and the changes were more obvious in Child-Pugh C group (P

DNA damage repair genes have been linked with increased aggressiveness of prostate cancer, however, the extent of mutation of these genes has not been analyzed within a cohort of African American patients. Here, the authors identify increased mutation rates in specific DNA repair genes, compared with prostate cancer patients with European Ancestry.

Bovine endometritis is characterized by reduced milk production and high rates of infertility. Prior research has indicated that melatonin may possess anti-inflammatory and antioxidant properties that can counteract the progression of inflammatory diseases. In this research, we attempted to elucidate the protective effects of melatonin on LPS-induced endometritis. The results obtained from enzyme-linked immunosorbent assay (ELISA) and quantitative real-time PCR (qRT-PCR) revealed that melatonin effectively reduced the production and release of pro-inflammatory cytokines in an LPS-induced bovine endometrial epithelial cell line (BEND cells). Furthermore, western blotting demonstrated that melatonin treatment reduced the expression levels of the NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome-related proteins, including NLRP3, activated caspase-1, and cleaved IL-1β. Importantly, we further demonstrated that the anti-inflammatory effect of melatonin on BEND cells was related to autophagy by western blotting. Moreover, we used western blotting to detect autophagy-related proteins, MitoSOX to detect mitochondrial reactive oxygen species production (mtROS), and mitochondrial membrane potential (MMP) assay to detect mitochondrial membrane potential. The administration of melatonin demonstrated a significant enhancement in autophagy within BEND cells, leading to the effective elimination of impaired mitochondria. This process resulted in a reduction in the generation of reactive oxygen species within the mitochondria, restoration of mitochondrial membrane potential, and inhibition of the NLRP3 inflammasome activation. Moreover, in a mouse model of LPS-induced endometritis, melatonin treatment repressed the expression of pro-inflammatory cytokines by ELISA and qRT-PCR, alleviated pathological changes by hematoxylin–eosin staining (H&E), and inhibited myeloperoxidase (MPO) activity. In conclusion, our study showed that melatonin inhibited the activation of the NLRP3 inflammasome in BEND cells through autophagy, which may provide a novel therapeutic strategy for bovine endometritis.

Abstract Background Microvascular insufficiency plays an important role in the development of diabetic cardiomyopathy (DCM), therapeutic angiogenesis has been mainly used for the treatment of ischemic diseases. This study sought to verify the preclinical performance of SonoVue microbubbles (MB) combined ultrasound (US) treatment on myocardial angiogenesis in the rat model of DCM and investigate the optimal ultrasonic parameters. Methods The male Sprague–Dawley (SD) rats were induced DCM by streptozotocin through intraperitoneal injecting and fed with high-fat diet. After the DCM model was established, the rats were divided into the normal group, DCM model group, and US + MB group, while the US + MB group was divided into four subsets according to different pulse lengths (PL) (8 cycles;18 cycle;26 cycle; 36 cycle). After all interventions, all rats underwent conventional echocardiography to examine the cardiac function. The rats were sacrificed and myocardial tissue was examined by histology and morphometry evaluations to detect the myocardial protective effect of SonoVue MBs using US techniques. Results From morphologic observation and echocardiography, the DCM rats had a series of structural abnormalities of cardiac myocardium compared to the normal rats. The US-MB groups exerted cardioprotective effect in DCM rats, improved reparative neovascularization and increased cardiac perfusion, while the 26 cycle group showed significant therapeutic effects on the cardiac functions in DCM rats. Conclusion This strategy using SonoVue MB and US can improve the efficacy of angiogenesis, even reverse the progress of cardiac dysfunction and pathological abnormalities, especially using the 26 cycle parameters. Under further study, this combined strategy might provide a novel approach for early intervention of DCM in diabetic patients.

Background. To explore the application value of layered strain technique in non-ST elevation acute coronary syndrome (NSTE-ACS). Methods. 120 patients with suspected NSTE-ACS undergoing coronary angiography in our hospital from December 2018 to December 2019 were prospectively selected. According to the results of coronary angiography, the patients were divided into the significant CAD group and the nonsignificant CAD group. Echocardiography was performed 1-2 hours before invasive coronary angiography. The long axis and circumferential strain of the endocardium, myocardial layer, and epicardium were evaluated by the layered strain technique. The territorial longitudinal strain (TLS), the global longitudinal strain (GLS) of the three myocardial layers, and the global circumferential strain (GCS) were calculated based on the perfusion region of the three coronary arteries and the 17-segment model of the left ventricle. The primary endopoints were TLS and GCS of the three-layer myocardium. Results. Compared with the nonsignificant CAD patients, the TLS and GCS of three-layer myocardium in significant CAD patients were decreased, especially in the endocardium. The absolute values of TLS and GCS of the endocardium and epicardium in significant CAD patients were lower than those in nonsignificant CAD patients. This indicates a significant decrease in endocardial function. Receiver operating characteristic (ROC) curve analysis showed that endocardial TLS was superior to LVEF, Troponin I (TnI), and other strain parameters in evaluating the extent of coronary lesions. Conclusions. The layered strain technique of 2D-STE can evaluate the severity of coronary lesions in patients with NSTE-ACS, and for significant CAD patients, endocardial function is significantly more impaired than epicardial function.

Hard petroleum asphalt, which is a high modulus asphalt binder material, is used for improving asphalt pavement rutting distress. This property is attributed to the good high-temperature rheological characteristics of hard petroleum asphalt. The study investigates the performance of hard petroleum asphalt and Styrene-Butadiene-Styrene polymer modified asphalt (SBSPMA) based on three aspects: asphalt oil source, asphalt grade, and aging degree. The short-term and long-term aging of four hard petroleum asphalts and SBSPMA of comparative samples were measured using the rolling thin film oven test (RTFOT) and the pressure aging test (PAV). Firstly, the viscosity-temperature curve of hard petroleum asphalt was obtained using the Brookfield viscosity test at different temperatures combined with the improved REFUTAS formula. The complex shear modulus (G∗), phase angle (δ), and rutting factor (G∗/sin δ) of each asphalt sample were then measured by the dynamic shear rheometer test (DSR). Furthermore, the temperature sensitivity of each asphalt sample was calculated using the complex modulus index method (GTS). The compaction and paving temperature ranges of each asphalt were then confirmed. The results showed that the high-temperature rheological characteristics of hard petroleum asphalt were similar to those of SBSPMA. The temperature sensitivity of hard petroleum asphalt was basically the same as that of SBSPMA, while the influence of aging on the G∗ of hard petroleum asphalt was greater than that of SBSPMA. In addition, the hard petroleum asphalt construction temperature was lower than that of SBSPMA.

To some extent, the photosynthetic traits of developing leaves of maize are regulated systemically by water and nitrogen. However, it remains unclear whether photosynthesis is systematically regulated via water and nitrogen when maize crops are grown under close (high density) planting conditions. To address this, a field experiment that had a split-split plot arrangement of treatments was designed. Two irrigation levels on local traditional irrigation level (high, I2, 4,050 m3 ha−1) and reduced by 20% (low, I1, 3,240 m3 ha−1) formed the main plots; two levels of nitrogen fertilizer at a local traditional nitrogen level (high, N2, 360 kg ha−1) and reduced by 25% (low, N1, 270 kg ha−1) formed the split plots; three planting densities of low (D1, 7.5 plants m−2), medium (D2, 9.75 plants m−2), and high (D3, 12 plants m−2) formed the split-split plots. The grain yield, gas exchange, and chlorophyll a fluorescence of the closely planted maize crops were assessed. The results showed that water–nitrogen coupling regulated their net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), quantum yield of non-regulated non-photochemical energy loss [Y(NO)], actual photochemical efficiency of PSII [Y(II)], and quantum yield of regulated non-photochemical energy loss [Y(NPQ)]. When maize plants were grown at low irrigation with traditional nitrogen and at a medium density (i.e., I1N2D2), they had Pn, Gs, and Tr higher than those of grown under traditional treatment conditions (i.e., I2N2D1). Moreover, the increased photosynthesis in the leaves of maize in the I1N2D2 treatment was mainly caused by decreased Y(NO), and increased Y(II) and Y(NPQ). The coupling of 20%-reduced irrigation with the traditional nitrogen application boosted the grain yield of medium density-planted maize, whose Pn, Gs, Tr, Y(II), and Y(NPQ) were enhanced, and its Y(NO) was reduced. Redundancy analysis revealed that both Y(II) and SPAD were the most important physiological factors affecting maize yield performance, followed by Y(NPQ) and NPQ. Using the 20% reduction in irrigation and traditional nitrogen application at a medium density of planting (I1N2D2) could thus be considered as feasible management practices, which could provide technical guidance for further exploring high yields of closely planted maize plants in arid irrigation regions.

Abstract Cystic fibrosis (CF) is a life‐limiting autosomal recessive genetic disease caused by variants in the CFTR gene, most commonly by the [F508del] variant. Although CF is a classical Mendelian disease, genetic variants in several modifier genes have been associated with variation of the clinical phenotype for pulmonary and gastrointestinal function and urogenital development. We hypothesized that whole genome sequencing of a well‐phenotyped CF populations might identify novel variants in known, or hitherto unknown, modifier genes. Whole genome sequencing was performed on the Illumina HiSeq X platform for 98 clinically diagnosed cystic fibrosis patient samples from the Adult CF Clinic at the University of California San Diego (UCSD). We compared protein‐coding, non‐silent variants genome wide between CFTR [F508del] homozygotes vs CFTR compound heterozygotes. Based on a single variant score test, we found 3 SNPs in common variants (MAF >5%) that occurred at significantly different rates between homozygous [F508del]CFTR and compound heterozygous [F508del]CFTR patients. The 3 SNPs were all located in one gene on chromosome 2: Tensin 1 (TNS1: rs3796028; rs2571445: and rs918949). We observed significantly lower BMIs in homozygous [F508del]CFTR patients who were also homozygous for Tensin 1 rs918949 (T/T) (p = 0.023) or rs2571445 (G/G) (p = 0.02) variants. The Tensin 1 gene is thus a potential modifier gene for low BMI in CF patients homozygous for the [F508del]CFTR variant.

Abstract Posttraumatic stress disorder is developed by exposure to a threatening and/or a horrifying event and characterized by the presence of anxiety, hyperarousal, avoidance, and sleep abnormality for a prolonged period of time. To elucidate the potential molecular mechanisms, we constructed a mouse model by electric foot shock followed by situational reminders and performed transcriptome analysis in brain tissues. The stressed mice acquired anxiety-like behavior after 2 weeks and exaggerated startle response after 4 weeks. Avoidance latency and freezing behavior were sustained up to 5 weeks post stress and abnormal static behavior was observed during the sleep period. RNA sequencing was performed in two of the emotional regulatory regions, anterior cingulate cortex and amygdala, at 2 and 5 weeks post stress. More than 1000 differentially expressed genes were identified at 2 weeks in both regions. The number of the regulated genes remained constant in amygdala at 5 weeks post stress, whereas those in anterior cingulate cortex were plummeted. Although synaptic remodeling and endocrine system were the most enriched signaling pathways in both anterior cingulate cortex and amygdala, the individual gene expression profile was regulated in a region- and time-dependent manner. In addition, several genes associated with PTSD involved in Hypothalamic-Pituitary-Adrenal axis were differentially regulated. These findings suggested that global gene expression profile was dynamically regulated in accordance with the disease development stage, and therefore targeting the distinct signaling molecules in different region and development stage might be critical for effective treatment to PTSD.

Polymer is a kind of high molecular elastic material. The polymer cement mortar composite material formed by mixing it with cement mortar has the advantages of light weight, high strength, and good durability compared with traditional mortar materials. The effect of polyurethane polymer content on mechanical properties and microstructure of polyvinyl alcohol (PVA) fiber cement mortar was studied by compressive test, flexural test, and SEM analysis. The test results show that as the content of polyurethane increases, the compressive strength gradually decreases, and the flexural strength gradually increases. The addition of polyurethane helps to optimize the microstructure of PVA mortar, improve the compactness of the material, and enhance the bending resistance of the mortar. The mechanical properties of materials obtained from the experiment can provide references for engineering applications.

The concentric pushing method was used to study the bonding properties between polymer and concrete. This paper studied the influence of polymer density, environmental temperature, and moisture content of concrete between polymer and concrete on the bond strength. The results indicated that the bond failure of specimens occurred mainly when the polymer was pushed out. Furthermore, increasing the polymer density increases the bond strength at the polymer-concrete interface but decreases as the moisture content of the concrete increases. The environmental temperature affects the curing time, and the bond strength increases with increasing temperature. Under the same condition, the bond strength was influenced by the roughness of the interface. This study provides references for the construction design and enhances polymer materials and matrix application for repairing cracks in concrete dams.

Castration Resistant Prostate Cancer (CRPC) is thought to be driven by a collaborative mechanism between TNFα/NFκB and TGFβ signaling, leading to inflammation, Epithelial-to-Mesenchymal-Transition (EMT), and metastasis. Initially, TGFβ is a tumor suppressor, but in advanced metastatic disease it switches to being a tumor promoter. TGFBR2 may play a critical role in this collaboration, as its expression is driven by NFκB and it is the primary receptor for TGFβ. We have previously reported that the cardenolide drug digitoxin blocks TNFα/NFκB-driven proinflammatory signaling. We therefore hypothesized that digitoxin might break the collaborative process between NFκB and TGFβ by also inhibiting expression of TGFBR2. We therefore tested whether TGFβ-driven EMT and resulting metastases would be suppressed. Here we show, in vitro, that digitoxin inhibits NFκB-driven TGFBR2 expression, as well as Vimentin, while elevating E-cadherin expression. Digitoxin also significantly reduces HSPB1 mRNA and the HSPB1/RBFOX2 mRNA ratio in PC3 cells. In vivo, in a syngeneic, immune competent rat model of metastatic CRPC, we show that digitoxin also suppresses Tgfbr2 expression, as well as expression of other genes classically driven by NFκB, and of multiple EMT genes associated with metastasis. Concurrently, digitoxin suppresses tumor growth and metastasis in these animals, and prolongs survival. Gross tumor recurrence following tumor resection also appears prevented in ca 30% of cases. While the existence of a collaboration between NFκB and TGFβ to drive EMT and metastasis has previously been appreciated, we show here, for the first time, that chronic, low concentrations of digitoxin are able to block CRPC tumor progression, EMT and the ensuing metastatic disease.

For unknown reasons, there is huge variability in risk conferred by different HPV types and, remarkably, strong differences even between closely related variant lineages within each type. HPV16 is a uniquely powerful carcinogenic type, causing approximately half of cervical cancer and most other HPV-related cancers. To permit the large-scale study of HPV genome variability and precancer/cancer, starting with HPV16 and cervical cancer, we developed a high-throughput next-generation sequencing (NGS) whole-genome method. We designed a custom HPV16 AmpliSeq™ panel that generated 47 overlapping amplicons covering 99% of the genome sequenced on the Ion Torrent Proton platform. After validating with Sanger, the current “gold standard” of sequencing, in 89 specimens with concordance of 99.9%, we used our NGS method and custom annotation pipeline to sequence 796 HPV16-positive exfoliated cervical cell specimens. The median completion rate per sample was 98.0%.Our method enabled us to discover novel SNPs, large contiguous deletions suggestive of viral integration (OR of 27.3, 95% CI 3.3–222, P=0.002), and the sensitive detection of variant lineage coinfections. This method represents an innovative high-throughput, ultra-deep coverage technique for HPV genomic sequencing, which, in turn, enables the investigation of the role of genetic variation in HPV epidemiology and carcinogenesis. Keywords: HPV16, HPV epidemiology, HPV genomics

Warm-mixed asphalt (WMA) has the characteristics of low mixing temperature and energy consumption, which makes it more suitable than hot-mixed asphalt for plateau regions (the altitude is above 500 m, and the terrain is relatively flat or has a wide area with certain undulations). However, WMA is subject to severe ultraviolet (UV) aging because the UV radiation in plateau areas is more intense. The asphalt’s aging changes its rheological properties inevitably, and thus degrades the asphalt pavement’s performance. Accordingly, the purpose of this paper is to investigate the effect of UV and oxygen coupling aging on WMA’s rheological properties and functional group index. Temperature and frequency sweep tests were performed with a dynamic shear rheometer. At the same time, the functional group index was used as an indicator to compare the changes in the WMA’s infrared spectrum before and after UV aging. The results showed that WMA’s elasticity increased and its viscosity decreased after aging with UV. Under the condition of UV–oxygen isolation, as the aging period increased, the WMA’s rutting factor increased gradually. The degree of improvement was greater than that under the condition of oxygen isolation. In contrast, the time required for the WMA’s complex shear modulus to recover under the condition of UV–oxygen coupling was delayed. An increase in the peak value of infrared spectrum indicates that the WMA has undergone oxygen aging. The presence and change in the carbonyl group reflects the degree of the WMA’s UV aging, and the UV aging condition had a greater effect on the peak values of the carbonyl and sulfoxide groups.

Following publication of the original article [1], the authors reported that one of the authors’ names was spelled incorrectly.

Hodgkin lymphoma shows strong familial aggregation but no major susceptibility genes have been identified to date. The goal of this study was to identify high-penetrance variants using whole exome sequencing in 17 Hodgkin lymphoma prone families with three or more affected cases or obligate carriers (69 individuals), followed by targeted sequencing in an additional 48 smaller HL families (80 individuals). Alignment and variant calling were performed using standard methods. Dominantly segregating, rare, coding or potentially functional variants were further prioritized based on predicted deleteriousness, conservation, and potential importance in lymphoid malignancy pathways. We selected 23 genes for targeted sequencing. Only the p.A1065T variant in KDR (kinase insert domain receptor) also known as VEGFR2 (vascular endothelial growth factor receptor 2) was replicated in two independent Hodgkin lymphoma families. KDR is a type III receptor tyrosine kinase, the main mediator of vascular endothelial growth factor induced proliferation, survival, and migration. Its activity is associated with several diseases including lymphoma. Functional experiments have shown that p.A1065T, located in the activation loop, can promote constitutive autophosphorylation on tyrosine in the absence of vascular endothelial growth factor and that the kinase activity was abrogated after exposure to kinase inhibitors. A few other promising mutations were identified but appear to be “private”. In conclusion, in the largest sequenced cohort of Hodgkin lymphoma families to date, we identified a causal mutation in the KDR gene. While independent validation is needed, this mutation may increase downstream tumor cell proliferation activity and might be a candidate for targeted therapy.

Neurofibromatosis type 1 (NF1) is an autosomal dominant, monogenic disorder of dysregulated neurocutaneous tissue growth. Pleiotropy, variable expressivity and few NF1 genotype-phenotype correlates limit clinical prognostication in NF1. Phenotype complexity in NF1 is hypothesized to derive in part from genetic modifiers unlinked to the NF1 locus. In this study, we hypothesized that normal variation in germline gene expression confers risk for certain phenotypes in NF1. In a set of 79 individuals with NF1, we examined the association between gene expression in lymphoblastoid cell lines with NF1-associated phenotypes and sequenced select genes with significant phenotype/expression correlations. In a discovery cohort of 89 self-reported European-Americans with NF1 we examined the association between germline sequence variants of these genes with café-au-lait macule (CALM) count, a tractable, tumor-like phenotype in NF1. Two correlated, common SNPs (rs4660761 and rs7161) between DPH2 and ATP6V0B were significantly associated with the CALM count. Analysis with tiled regression also identified SNP rs4660761 as significantly associated with CALM count. SNP rs1800934 and 12 rare variants in the mismatch repair gene MSH6 were also associated with CALM count. Both SNPs rs7161 and rs4660761 (DPH2 and ATP6V0B) were highly significant in a mega-analysis in a combined cohort of 180 self-reported European-Americans; SNP rs1800934 (MSH6) was near-significant in a meta-analysis assuming dominant effect of the minor allele. SNP rs4660761 is predicted to regulate ATP6V0B, a gene associated with melanosome biology. Individuals with homozygous mutations in MSH6 can develop an NF1-like phenotype, including multiple CALMs. Through a multi-platform approach, we identified variants that influence NF1 CALM count.