35, 95% confidence interval, 1.35-4.09, p = 0.003) were associated with post-HCT PC utilization. Within one-year of hematopoietic cell transplantation, 127 patients died; among those, recipients of early PC consultation had significantly higher rates of hospice enrollment (25% vs. https://www.selleckchem.com/MEK.html 9%, p = 0.019) and lower rates of hospital death (71% vs. 90%, p = 0.013), intensive care unit admission (44% vs. 75%, p = 0.001), and high-intensity medical care in last 30 days of life (46% vs. 77%, p = 0.001). Conclusions Our results highlight important pre-HCT risk factors associated with increased PC needs posthematopoietic cell transplantation and benefits of PC involvement for older allo-HCT recipients at the end of life. Prospective studies should examine the optimal timing of PC consultation and its multidimensional benefits for older allo-HCT patients.Purpose Low dose radiation has been reported as an effective treatment for rheumatoid arthritis via multiple dose exposures. The present study was designed to increase the therapeutic efficacy of low dose radiation with the minimum exposure level in arthritic rats by concurrent administration of resveratrol (RSV) as an adjunctive therapy with anti-inflammatory properties.Materials and methods Rats were rendered arthritic by sub-plantar injection of Freund's complete adjuvant (FCA) and exposed to low dose radiation at a total exposure level of 0.5 Gy (2 x 0.25). During the exposure course, RSV (50 mg/kg) was orally administered once daily for two weeks. Diclofenac (3 mg/kg) was administered as a standard anti-inflammatory drug. Paw volume was measured every 4 days. After 28 days of induction, rats were sacrificed and serum was collected for estimation of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), thiobarbituric acid reactive substances (TBARS), and total nitrate/nitrite (NOx). Furthermore, paws were dissected for histopathological examinations and immuno-histochemical estimation of nuclear factor-kappa B p65 (NF-κB p65) expression.Results Administration of RSV during the low dose radiation exposure course produced a significant decrease in the paw swelling and a potentiated inhibition in the serum levels of TNF-α, IL-1β, TBARs, and NOx. The dual treatment strategy alleviated the histopathological damage to a greater extent than that produced by each treatment. Moreover, a pronounced suppression of NF-κB p65 expression in the synovial tissue was observed in the combination group. The combination treatment showed a nearly similar potency to that observed in the diclofenac treated group.Conclusion Administration of RSV augmented the modulatory activity of low dose radiation with minimum exposure level on the disease progression.The study aims to establish how feasible a natural therapy option (safflower oil) is in the treatment of postoperative pain. Naproxen sodium has already been experimentally proven to be effective for this purpose. Accordingly, the analgesic and anti-inflammatory effects of safflower oil were compared with those obtained with benzydamine HCl and naproxen sodium. Forty-two, healthy, adult female rats of Wistar albino species were divided at random into six groups of seven rats. The intervention allocation was as follows Group No. 1-physiological saline 0.9%; Group No. 2-safflower oil 100 mg/kg; Group No. 3-safflower oil 300 mg/kg; Group No. 4-benzydamine HCl 30 mg/kg; Group No. 5-benzydamine HCl 100 mg/kg; and Group No. 6-naproxen sodium 10 mg/kg. Following allocation of treatment, pain was induced experimentally and tested in various ways (hot plate test, tail-pinching test, and writhing test) and the efficacy of each treatment in providing peripheral and central analgesia was evaluated. The second stage consied in venous congestion between placebo and safflower oil 300 mg/kg; in damage to the epithelium between placebo and safflower oil 300 mg/kg and between naproxen sodium 10 mg/kg and safflower oil; and in cell infiltration and development of edema between placebo and safflower oil 300 mg/kg. It is predicted that further research into safflower oil and benzydamine HCl will create opportunities to develop analgesic-anti-inflammatory therapeutics of a novel kind for the treatment of postoperative pain and inflammation.Background Caregivers are decision stakeholders; yet, few interventions have been developed to help patients and caregivers collaborate on advance care planning (ACP). Objective To evaluate a theory-based ACP pilot intervention, Deciding Together, to improve decisional quality, readiness, collaboration, and concordance in ACP decisions for older adult home health (HH) patients and caregivers. Design A one-group, pre- and posttest study using matched questionnaires was conducted. The intervention consisted of a clinical vignette, theoretically guided conversation prompts, and a shared decision-making activity. Setting/Subjects N = 36 participants (n = 18 HH patients; n = 18 family and nonfamily caregivers) were purposively recruited from a HH agency to participate in the intervention at patients' homes. Measurements Demographic and baseline measures were collected for relationship quality, health status, and previous ACP engagement. Outcome measures included perceptions of collaboration, readiness for ACP, concordance in life-sustaining treatment preferences (cardiopulmonary resuscitation, antibiotics, artificial nutrition and hydration, and mechanical ventilation), and decisional conflict. Descriptive statistics, Cohen's κ coefficients, paired t tests, McNemar's tests, and Wilcoxon signed-rank tests (and effect size estimates, r = z/√N) were calculated using R-3.5.1 (p  less then  0.05). Single value imputation was used for missing values. Results While no significant differences were found for perceptions of collaboration, and readiness for ACP, patients (r = 0.38, p = 0.02) and caregivers (r = 0.38, p = 0.02) had reduced decisional conflict at posttest. Patients' and caregivers' agreement increased by 27.7% for an item assessing patients' preference for artificial nutrition and hydration (p = 0.03). Conclusions This study suggests that collaborative ACP decision making may improve decisional conflict for older adult HH patients and their caregivers.

BACKGROUND We sought to investigate the expression of KPNA2 in bladder cancer (BC) and its relationship with prognosis, and to analyze the potential mechanism of KPNA2 in promoting BC progression. MATERIAL AND METHODS The RNA-seq data on BC from The Cancer Genome Atlas (TCGA) database were imported into R statistical software for differential analysis. The clinical data for patients with BC were screened and analyzed with R software. The survival curve was drawn with the Kaplan-Meier Plotter. The expression of KPNA2 in 4 human BC cell lines and a human bladder epithelial cell line was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting (WB). The proliferation of BC cells was detected with Cell Counting Kit-8 (CCK8), detection of apoptosis, and flow cytometry, and the migration and invasion of BC cells were detected through Transwell assays. WB was used to detect proteins involved in the P53 pathway. RESULTS The expression of KPNA2 was higher in BC. The difference in KPNA2 expression was associated with many clinicopathological factors, and high expression of KPNA2 was associated with shorter survival time. After KPNA2 knockout, the proliferation, migration, and invasion ability decreased significantly, the cell cycle was clearly arrested in the G0/G1 phase, and the number of apoptotic cells increased. Moreover, CyclinD1, BCL2, and pro-caspase3 decreased significantly, whereas P53, P21, BAX, and cleaved-caspase3 increased significantly. The results in the overexpression group were the opposite of results in the knockdown group. CONCLUSIONS KPNA2 is an oncogenic factor that facilitates BC tumorigenicity through the P53 pathway.BACKGROUND Cefepime-induced neurotoxicity has been described in intensive care units (ICUs) and neuro ICU settings, occurring in patients started on cefepime for management of severe infections and sepsis. Most cases occur within 1 to 10 days after starting the drug. https://www.selleckchem.com/Androgen-Receptor.html We publish a case that occurred on the general medical ward of a patient who had been on cefepime therapy for 4 weeks prior to admission. The aim of this study was to improve the knowledge of this serious condition to general internists as our patient was being managed on the general medical ward. CASE REPORT A 72-year-old female on prolonged intravenous antibiotics for sacral and pelvic osteomyelitis presented with acute encephalopathy and aphasia in the setting of an acute kidney injury. Due to the acute focal neurologic deficit, she was initially admitted as a stroke alert. After a negative magnetic resonance imaging (MRI) of the brain, an electroencephalogram (EEG) was pursued and showed nonconvulsive status epilepticus (NCSE). NCSE was likely a result of cefepime therapy in the setting of an acute kidney injury. CONCLUSIONS Cefepime-induced neurotoxicity should be suspected in any patient on cefepime therapy who develops acute changes in mental status, myoclonus, or evidence of seizures. Risk factors for the disease include older age, renal dysfunction, critical illness, and inappropriate dosing based upon renal function. A high index of suspicion is required and delays in diagnosis are common as there are frequently multiple possible causes for altered mental status in systemically ill patients requiring treatment with broad-spectrum antibiotics.Brugada syndrome (BrS) is a congenital channelopathy associated with the development of malignant ventricular arrhythmias and sudden cardiac death. The diagnosis of BrS is made based on Brugada ECG pattern and clinical history. Brugada phenocopies (BrP) are clinical entities that are characterized by ECG patterns identical to those of BrS but arise from different underlying conditions such as metabolic abnormalities, myocardial ischemia and mechanical compression. Distinction between the two is important because BrS requires investigations for risk stratification whereas BrP requires appropriate treatment for the underlying conditions. In the present review, we summarized the current data in the literature, systematic diagnostic approach, gaps in the literature and future perspective on BrP.OBJECTIVE A permanent pacemaker (PPM) is necessary for patients with a symptomatic third-degree or advanced second-degree atrioventricular (AV) block. An AV block due to medication use can often be reversed; however, subsequent relapse can occur and necessitate subsequent PPM implantation. The aim of this study was to explore the course and prognosis of patients with an AV block. METHODS This historical cohort study was conducted between January 2013 and June 2018. A total of 1900 patient records were analyzed and 1123 subjects with an AV block on admission were enrolled. The patients were categorized into 2 groups Group 1 comprised patients with an AV block due to medication use (n=316, 28%) and Group 2 included patients with an AV block caused by other etiologies (n=807, 72%). Data of the cause of AV block, recurrence, and PPM implantation were analyzed. Patients in both groups who did not require a PPM during the index admission were followed up regarding subsequent implantation of a PPM. RESULTS AV conduction was recovered in 38 (12%) patients in Group 1 and 48 (6%) patients in Group 2 during the index hospitalization. However, recurrence of the AV block was observed in 18% of Group 1 patients and 40% of Group 2 patients. Only 25 patients in each group (4.5% of the whole study population) remained PPM-free during a median 3-year follow-up period. CONCLUSION The study findings suggest that drug-induced AV blocks may not be as benign as previously thought. The high relapse rate indicates that watchful follow-up may be required despite discontinuation of the responsible medication and that consideration of earlier PPM implantation in cases of early recurrence may be warranted.OBJECTIVE Inappropriate sinus tachycardia (IST) is a syndrome characterized by an elevated resting heart rate with distressing symptoms and no secondary cause of sinus tachycardia. This study was conducted to evaluate both the prevalence of IST among symptomatic patients and heart rate variability (HRV) characteristics. METHODS The records of all consecutive symptomatic patients who had undergone 24-hour Holter monitoring between September 2015 and November 2016 at a single center were retrospectively evaluated. IST was defined as a 24-hour mean heart rate (HR) of ≥90 beats/minute and a resting HR of ≥100 beats/minute in the absence of any secondary cause of sinus tachycardia. All of the study data related to clinical characteristics, symptoms, concomitant diseases, and Holter electrocardiogram parameters were obtained from the electronic hospital records. A propensity age- and sex-matched control group was selected from a non-IST patient cohort. RESULTS A total of 1865 consecutive patients were evaluated and 32% were excluded due to an inadequate Holter recording period or insufficient quality, atrial fibrillation episodes, atrioventricular block, or >1% atrial or ventricular extrasystoles.

CRISPR/Cas9-mediated editing of Clpsk1 enhanced watermelon resistance to Fusarium oxysporum. The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system has proven to be an effective genome-editing tool for crop improvement. Previous studies described that Phytosulfokine (PSK) signalling attenuates plant immune response. In this work, we employed the CRISPR/Cas9 system to knockout Clpsk1 gene, encoding the PSK precursor, to confer enhanced watermelon resistance to Fusarium oxysporum f.sp. niveum (FON). Interactions between PSK and FON were analysed and it was found that transcript of Clpsk1 was significantly induced upon FON infection. Meanwhile, application of exogenous PSK increased the pathogen growth. Then, one sgRNA, which targeted the first exon of Clpsk1, was selected for construction of pRGEB32-CAS9-gRNA-Clpsk1 expression cassette. The construct was then transformed to watermelon through Agrobacterium tumefaciens-mediated transformation method. Six mutant plants were obtained and three types of mutations at the expected position were identified based on Sanger sequencing. Resistance evaluation indicated that Clpsk1 loss-of-function rendered watermelon seedlings more resistant to infection by FON. These results indicate that CRISPR/Cas9-mediated gene modification is an effective approach for watermelon improvement.Twenty-three PeLACs have been identified in moso bamboo, overexpression of PeLAC10 increases the lignin content and confers drought and phenolic acid tolerance in transgenic Arabidopsis. Laccases (LACs) have multifunction involved in the processes of cell elongation, lignification and stress response in plants. However, the function of laccases in bamboo remain unclear. Here, a total of 23 laccase genes (PeLAC1-PeLAC23) were identified in moso bamboo (Phyllostachys edulis). The diverse gene structure and expression pattern of PeLACs suggested that their function should be spatiotemporal and complicated, which was supported by the expression profiles in different tissues of moso bamboo. Eighteen PeLACs were identified as the targets of ped-miR397. The putative ped-miR397-binding site in the coding region of PeLAC10 was further confirmed by RLM-5' RACE, indicating that PeLAC10 was regulated by ped-miR397 after transcription. With the increasing shoot height, the expression abundance of PeLAC10 was up-regulated and reached the maximum in 15 cm shoots, while that of ped-miR397 was relative lower and showed the minimum in 15 cm shoots. PeLAC10 was up-regulated obviously under both ABA (100 μmol L-1) and NaCl (400 mmol L-1) treatments, and it was down-regulated under the GA3 (100 μmol L-1) treatment. The transgenic Arabidopsis plants over-expressing PeLAC10 became slightly smaller and their petioles were shorter than those of Col-0. However, they had a stronger capacity in resistance to phenolic acids and drought besides higher lignin content in stems. https://www.selleckchem.com/TGF-beta.html These results indicated that overexpression of PeLAC10 was helpful to increase the content of lignin in transgenic Arabidopsis and improve the adaptability to phenolic acid and drought stresses.The use of homeopathy is remarkably popular. Popularity, however, is not an arbiter in a scientific discourse. In fact, the assumptions underlying homeopathy violate fundamental laws of nature. Homeopathy does not have any explanatory power and fails other criteria established for a scientific approach. Two large-scale efforts have recently documented that in spite of a plethora of clinical trials there is no evidence that homeopathic remedies have any therapeutic effect, which goes beyond that of a placebo. Relaxed regulations and lack of scientific literacy and of health education allow for continuous thriving of homeopathy. While the tide may be changing on the regulatory side, health education of the general public is presumably more important to support informed decision making by patients. Otherwise, the responsible patient, who is posited to decide on the medical choices, remains a convenient legal fiction.OBJECTIVE To compare the application effects of venous indwelling needles and intravenous infusion with steel needles in pediatric outpatient care and to observe the complications of the two approaches. METHODS A total of 110 children who received intravenous infusions in our hospital from March 2018 to May 2018 were selected as study subjects. Children were assigned to an observation group (n = 55) or a control group (n = 55) based on a random number table, of which the children in the control group received intravenous infusion with the steel needles while the children in the observation group received intravenous infusion with indwelling needles. Their mental states were rationally evaluated by the state-trait anxiety inventory (STAI) and modified Yale preoperative anxiety score (MYPAS) in the two groups. At the same time, the total number of punctures in 3 days of infusion, complications and the parents' satisfaction with nursing service were compared between the two groups. RESULTS The difference in STAI and MYPAS scores was statistically significant between the two groups (P  0.05) between the two groups, but the difference in the total number of punctures on the second day and the third day was statistically significant (P  less then  0.05). Besides this, the incidence of complications in the observation group was less than that in the control group, and the nursing satisfaction in the observation group was higher than that in the control group (P  less then  0.05). CONCLUSION The children's psychological impacts are relatively small when comparing the venous indwelling needle with the steel needle. Moreover, venous indwelling needles avoid repeated punctures; therefore, the parents' satisfaction in nursing is improved, and it is worth popularizing.PURPOSE To compare the effect of the lateral meniscus (LM) complete radial tear at different tear sites on the load distribution and transmission functions. METHODS A compressive load of 300 N was applied to the intact porcine knees (n = 30) at 15°, 30°, 60°, 90°, and 120° of flexion. The LM complete radial tears were created at the middle portion (group M), the posterior portion (group P), or the posterior root (group R) (n = 10, each group), and the same loading procedure was followed. Finally, the recorded three-dimensional paths were reproduced on the LM-removed knees. The peak contact pressure (contact area) in the lateral compartment and the calculated in situ force of the LM under the principle of superposition were compared among the four groups (intact, group M, group P, and group R). RESULTS At all the flexion angles, the peak contact pressure (contact area) was significantly higher (lower) after creating the LM complete radial tear as compared to that in the intact state (p  less then  0.01). At 120° of flexion, group R represented the highest peak contact pressure (lowest contact area), followed by group P and group M (p  less then  0.

Biochar is a "green" material that has been widely used in environmental applications for its capability to remove or immobilize contaminants in different environmental media (i.e. soil, water and air) and mitigate climate change. In this study, the feasibility of using KOH enhanced biochar for soil Cd and Pb stabilization was investigated, and the effects of pyrolysis temperature and alkaline concentrations for modification were explored. Field-emission scanning electron microscopy (FESEM), N2 adsorption-desorption, and Fourier Transform Infrared Spectroscopy (FTIR) analyses were conducted to reveal the influence on biochar physiochemical properties. The immobilization performances were examined through Toxicity Characteristics Leaching Procedure (TCLP), and Response Surface Methodology (RSM) was adopted to visualize the results from leaching tests. The stabilization mechanisms of alkaline enhanced biochars were investigated using Time of Flight Secondary Ion Mass Spectroscopy (TOF-SIMS), Tessier sequential extraction method and X-ray diffraction (XRD) analyses. The results indicated that rice husk biochar pyrolyzed at a relatively low temperature (i.e., 300 °C) and activated by moderate alkaline concentrations (i.e., 1 M or 3 M KOH) rendered optimum stabilization performance. KOH activation was a double-edged sword, with high alkaline concentrations destroying biochar's cell structures. Moreover, the integration of TOF-SIMS, XRD and sequential leaching method shed lights on the underlying mechanisms involved in metal stabilization. Surface complexation between toxic metals and oxygen-containing functional groups rather than liming or precipitation was proven to be the fundamental stabilization mechanism. The eastern Tibetan Plateau geothermal belt in the southwest of China hosts a number of hot springs with a wide range of temperature and hydrogeochemical conditions, which may harbor different niches for the distribution of microbial communities. In this study, we investigated hydrochemical characteristics and microbial community composition in 16 hot springs with a temperature range of 34.6 to 88.2 °C within and across three typical hydrothermal fields (Kangding, Litang, and Batang). According to aquifer lithologic and tectonic differences, the hydrochemical compositions of hot springs displayed an apparent regional-specific pattern with distinct distributions of major and trace elements (e.g., Ca2+, Mg2+, F-/B) and were primarily formed by water-rock interaction across the three hydrothermal fields. Nonetheless, microbial communities significantly assembled with the temperature rather than the geographic locations with distinct hydrogeological features. Low temperature (80% negative associations hinting a low co-existence pattern and highlighted the driving force of temperature as well as F- or total organic carbon (TOC) for microbial interactions. Microbial dissimilarity displayed significant linear correlations with environmental (temperature) and geographic distance in Batang but only with temperature in Kangding area, which might be attributed to the regional-specific hydrogeochemistry. This study may help us to better understand the distribution of the microbial community in hot spring across different hydrothermal fields. Developmental exposure to bisphenol A (BPA) is associated with liver dysfunction and diseases in adulthood. The aims of this study were to assess the effects of prenatal BPA exposure on the hepatic transcriptome and proteome in female and male offspring and to understand adverse outcome pathways (AOPs) to observed phenotypic effects. Pregnant Wistar rats were exposed to 50 or 5000 μg BPA/kg bw/day, or 17β-estradiol (E2, 50 μg/kg bw/day) from embryonic day 3 to 18. The liver transcriptome and proteome profiles were analyzed in the newborn (postnatal day 1; PND1) and weaning (PND21) rat offspring. Based on the differentially expressed genes/proteins derived from transcriptome and proteome profiles, we performed pathway, transcription factor, and disease enrichment analyses. A principal component analysis of transcriptome data demonstrated that prenatal BPA exposure caused masculinization of the hepatic transcriptome in females. Both of transcriptomic and proteomic data showed that prenatal BPA exposure led to the disruption of cell cycle, lipid homeostasis, and hormone balance in offspring. Most of the effects at the transcript level were extended from newborn to weaning in males, but were moderated until weaning in females. The alterations at the transcript and protein levels were accordant with the observation of increases in body weight and anogenital distance and changes in hepatosomatic index in the offspring. Collectively, we constructed AOPs with evidence of sex- and age-specific actions of prenatal BPA exposure in the offspring. Although multiple sclerosis (MS) has been the leading cause of neurologically-induced disability in young adults, risk factors for the relapse and acute aggravation of MS remain unclear. A few studies have suggested a possible role of temperature changes on the relapse and acute aggravation of MS. We investigated the association between short-term exposure to wide diurnal temperature ranges (DTRs) and acute exacerbation of MS requiring an emergency department (ED) visit. A total of 1265 patients visited EDs for acute aggravation of MS as the primary disease in Seoul between 2008 and 2014 from the national emergency database. We conducted a conditional logistic regression analysis of the time-stratified case-crossover design to compare DTRs on the ED visit days for MS and those on control days matched according to the day of the week, month, and year. We examined possible associations with other temperature-related variables (ambient temperature, between-day temperature change, and sunlight hours). Short-term exposure to wide DTRs immediately increased the risk of ED visits for MS. https://www.selleckchem.com/products/p62-mediated-mitophagy-inducer.html Especially, 2-day average (lag0-1) DTR levels on the day of and one day prior to ED visits exhibited the strongest association (an 8.81% [95% CI 3.46%-14.44%] change in the odds ratio per 1 °C increase in the DTR). Other temperature-related variables were not associated with MS aggravation. Our results suggest that exposure to wider DTR may increase the risk of acute exacerbation of MS. Given the increasing societal burden of MS and the increasing temperature variability due to climate change, further studies are required.

BACKGROUND A step change in the night environment is taking place, with the large-scale installation of bright, broad-spectrum road lighting such as white light-emitting diodes (LEDs). One justification for this is a reduction in road traffic collisions (RTCs). This study aimed to estimate the effect of new lighting on personal injury RTCs within a large UK city. METHODS We analysed a 9-year time series of weekly RTC personal injury counts in 132 areas of the city using multilevel modelling. The RTC rate over a full 24-hour period was the primary outcome; darkness and daylight RTC rates were secondary. The background change in RTC rate was separated from the change associated with the number of newly installed bright lamps by including a polynomial underlying time trend for the logarithm of the mean number of collisions per week for each area. The study was based on a rigorous, predesigned and archived protocol. RESULTS Within-area coefficients for the broad lighting effect were positive; as the number of bright lamps in an area increased, so did the RTC rate. The estimate for the increase in the within-area 24-hour RTC rate is 11% (95% CI 2% to 20%). The estimate of darkness-only RTCs is 16% (95% CI 2% to 32%). If the effect of lighting on darkness RTC rate is adjusted by that for daylight, one obtains 4% (95% CI -12% to +23%). CONCLUSION No evidence was found for bright lamps leading to an improvement in road safety in any of the analyses. For this city, introducing brighter road lighting may have compromised safety rather than reducing harm. © Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY. Published by BMJ.Oxalate is a common constituent of kidney stones but the mechanism of its transport across epithelia are not well understood. With prior research on the role of the intestine focused on mammals this study considered oxalate handling by teleost fish. Given the osmotic challenge of seawater (SW), teleosts have limited scope for urinary oxalate excretion relative to freshwater (FW). The marine teleost intestine was hypothesized as the principal route for oxalate elimination thus demanding epithelial secretion. To test this, intestinal 14C-oxalate flux was compared between FW- and SW-acclimated sailfin molly (Poecilia latipinna). In SW, oxalate was secreted at remarkable rates (367.90±22.95 pmol cm-2 h-1) which were similar following FW transfer (387.59±27.82 pmol cm-2 h-1), implying no regulation by salinity. Nevertheless, this ability to secrete oxalate 15-19 times higher than mammalian small intestine supports this proposal of the teleost gut as a previously unrecognized excretory pathway. © 2020. Published by The Company of Biologists Ltd.PURPOSE ROS1 tyrosine kinase inhibitors (TKIs) provide significant benefit in lung adenocarcinoma (LUAD) patients with ROS1 fusions. However, as observed with all targeted therapies, resistance arises. Detecting mechanisms of acquired resistance (AR) is crucial to finding novel therapies and improve patient outcomes. EXPERIMENTAL DESIGN ROS1 fusions were expressed in in HBEC and NIH-3T3 cells either by cDNA overexpression (CD74/ROS1, SLC34A2/ROS1) or CRISPR-Cas9-mediated genomic engineering (EZR/ROS1). We reviewed targeted large-panel sequencing data (using the MSK-IMPACT assay) patients treated with ROS1 TKIs, and genetic alterations hypothesized to confer AR were modeled in these cell lines. RESULTS Eight of the 75 patients with a ROS1 fusion had a concurrent MAPK pathway alteration and this correlated with shorter overall survival. In addition, the induction of ROS1 fusions stimulated activation of MEK/ERK signaling in comparison with AKT signaling, suggesting the importance of the MAPK pathway in driving ROS1 fusion-positive cancers. Of 8 patients, 2 patients harbored novel in-frame deletions in MEK1 (MEK1delE41_L54) and MEKK1 (MEKK1delH907_C916) that were acquired after ROS1-TKIs, and 2 patients harbored NF1 loss-of-function mutations. Expression of MEK1del or MEKK1del, and knockdown of NF1 in ROS1 fusion-positive cells activated MEK/ERK signaling and conferred resistance to ROS1-TKIs. Combined targeting of ROS1 and MEK inhibited growth of cells expressing both ROS1 fusion and MEK1del. CONCLUSIONS We demonstrate that the activation of MAPK pathway is mechanisms of innate or acquired resistance and that patients harboring ROS1 fusion and concurrent MAPK alterations have worse survival. Our findings suggest a treatment strategy to target both aberrations. Copyright ©2020, American Association for Cancer Research.PURPOSE RNA splicing is a fundamental biological process that generates protein diversity from a finite set of genes. Recurrent somatic mutations of splicing factor genes are common in some hematological cancers but are relatively uncommon in Acute Myeloid Leukemia (AML, less then 20% of patients). https://www.selleckchem.com/products/n-formyl-met-leu-phe-fmlp.html We examined whether RNA splicing differences exist in AML, even in the absence of splicing factor mutations. EXPERIMENTAL DESIGN We developed a bioinformatics pipeline to study alternative RNA splicing in RNA-seq data from large cohorts of AML patients. RESULTS We have identified recurrent differential alternative splicing between patients with poor and good prognosis. These splicing events occurred even in patients without any discernible splicing factor mutations. Alternative splicing recurrently occurred in genes with specific molecular functions, primarily related to protein translation. Developing tools to predict the functional impact of alternative splicing on the translated protein, we discovered that ~45% of the splicing events directly affected highly conserved protein domains. Several splicing factors were themselves misspliced and the splicing of their target transcripts were altered. Studying differential gene expression in the same patients, we identified that alternative splicing of protein translation genes in ELNAdv patients resulted in the induction of an integrated stress response and up-regulation of inflammation-related genes. Lastly, using machine learning techniques, we identified a splicing signature of four genes which refine the accuracy of existing risk prognosis schemes and validated it in a completely independent cohort. CONCLUSIONS Our discoveries therefore identify aberrant alternative splicing as a molecular feature of adverse AML with clinical relevance. Copyright ©2020, American Association for Cancer Research.

Furthermore, strong correlations between metabolite types and bacterial genus were detected. There were remarkable differences in the gut microbiota composition and metabolomic profile between wild and captive Chinese monals. This study has established a baseline for a normal gut microbiota and metabolomic profile for wild Chinese monals, thus allowing us to evaluate if differences seen in captive organisms have an impact on their overall health and reproduction. There were remarkable differences in the gut microbiota composition and metabolomic profile between wild and captive Chinese monals. This study has established a baseline for a normal gut microbiota and metabolomic profile for wild Chinese monals, thus allowing us to evaluate if differences seen in captive organisms have an impact on their overall health and reproduction.Left ventricular thrombus (LVT) is associated with a significant risk of ischemic stroke (IS) and peripheral embolization. Societal guidelines recommend the use of warfarin, with direct oral anticoagulants (DOACs) only for patients unable to tolerate warfarin. We studied the natural history of LVT with anticoagulation (AC) with emphasis on comparing warfarin and DOAC use. In this single center study, we identified patients with a confirmed LVT. Type and duration of anticoagulation, INR levels and clinical outcomes (bleeding, ischemic stroke or peripheral embolization, and thrombus resolution) were recorded. LVT was confirmed in a total of 110 patients. https://www.selleckchem.com/products/PLX-4032.html Mean age was 59 + 14 years. 79% were men. Underlying etiology was chronic ischemic cardiomyopathy in 58%, non-ischemic cardiomyopathy in 23%. AC was started in 96 (87%) patients. At 1 year follow up, 11 patients (10%) had a stroke while on any AC (2 had hemorrhagic stroke and 9 had IS). Of those with IS, 7 were on warfarin (71% of those had subtherapeutic INR) and 2 patients on DOACs had IS. The 1-year risk of any stroke was 15% in warfarin group (12% risk of ischemic stroke) compared to 6% in the DOACs group (p = 0.33). 37 (63%) patients on warfarin and 18 (53%) on DOACs had resolution of thrombus (p = 0.85). One-year risk of stroke with LVT is high (10%) even with AC. Most patients IS on warfarin had subtherapeutic INR. There was no statistical difference in stroke risk or rate of thrombus resolution between warfarin and DOACs treated patients. Dietary nutrient intake plays a significant role in carcinogenesis. Few studies have investigated the association between dietary nutrient intake and cervical intraepithelial neoplasia (CIN) risk in China. Data on 2304 women from an ongoing cohort comprising 40,000 women from China in 2014 were included. Study randomly selected 218 out of 2304 people as subjects during 2019. All participants were surveyed through in-person interviews, physical examinations, and laboratory tests. Clinical data were obtained from physical examinations and laboratory tests. Dietary intakes were assessed using a semiquantitative food frequency questionnaire. Nutrition intakes from 26 food sources were calculated using a comprehensive validated database. Descriptive statistics were used to describe the frequency and proportion, and mean and standard deviation of the demographic characteristics. Characteristics were examined for significant differences, and Pearson chi-square tests were used for categoric variables. Logistic rerial Register (ChiCTR-ROC-15006479) ( https//www.chictr.org.cn ). Low folate; vitamin B6, C, and K; and niacin intakes were associated with CIN2+ risk. Nutrients may influence the development of higher grade CIN and cervical cancer. Trial registration The study was registered in the Chinese Clinical Trial Register (ChiCTR-ROC-15006479) ( https//www.chictr.org.cn ). The objective of this study was to develop a new predictive equation of resting energy expenditure (REE) for acute kidney injury patients (AKI) on dialysis. A cross-sectional descriptive study was carried out of 114 AKI patients, consecutively selected, on dialysis and mechanical ventilation, aged between 19 and 95years. For construction of the predictive model, 80% of cases were randomly separated to training and 20% of unused cases to validation. Several machine learning models were tested in the training data linear regression with stepwise, rpart, support vector machine with radial kernel, generalised boosting machine and random forest. The models were selected by ten-fold cross-validation and the performances evaluated by the root mean square error. There were 364 indirect calorimetry measurements in 114 patients, mean age of 60.65 ± 16.9years and 68.4% were males. The average REE was 2081 ± 645kcal. REE was positively correlated with C-reactive protein, minute volume (MV), expiratory positive airway pressure, serum urea, body mass index and inversely with age. The principal variables included in the selected model were age, body mass index, use of vasopressors, expiratory positive airway pressure, MV, C-reactive protein, temperature and serum urea. The final r-value in the validation set was 0.69. We propose a new predictive equation for estimating the REE of AKI patients on dialysis that uses a non-linear approach with better performance than actual models. We propose a new predictive equation for estimating the REE of AKI patients on dialysis that uses a non-linear approach with better performance than actual models. The feeding apparatus of salamanders consists mainly of the cranium, mandible, teeth, hyobranchial apparatus and the muscles of the cranial region. The morphology of the feeding apparatus in turn determines the boundary conditions for possible food processing (i.e., intraoral mechanical reduction) mechanisms. However, the morphology of the feeding apparatus changes substantially during metamorphosis, prompting the hypothesis that larvae might use a different food processing mechanism than post-metamorphic adults. Salamandrid newts with facultative metamorphosis are suitable for testing this hypothesis as adults with divergent feeding apparatus morphologies often coexist in the same population, share similar body sizes, and feed on overlapping prey spectra. We use high-speed videography to quantify the in vivo movements of key anatomical elements during food processing in paedomorphic and metamorphic Alpine newts (Ichthyosaura alpestris). Additionally, we use micro-computed tomography (μCT) to analyze morphological differences in the feeding apparatus of paedomorphic and metamorphic Alpine newts and sort them into late-larval, mid-metamorphic and post-metamorphic morphotypes.

7 mW and 0.64 nJ, respectively. During the experiment, the mode-locked pulse remained stable till the pump power reached a value of 400 mW and, for the regulation of power, the slope efficiency is calculated at about 4.99%. These results indicate that PdS2 material is a promising nonlinear optical material for ultrafast optical applications in the near-infrared (NIR) region.This study was performed to develop an adjuvant therapy in the form of a self-administered vaginal tablet regimen for the localized delivery of chemotherapeutic drugs. This therapy will help to reduce relapse by eradicating cancerous cells in the margin of cervical tumors. The vaginal tablet is a very common formulation that is easy to manufacture, easy to place in the vagina, and has a low cost of manufacture, making them ideal for use in developing countries. A combination of disulfiram and 5-fluorouracil, which are both off-patent drugs and provide different modes of action, were evaluated. The tablets developed were evaluated for weight variation, thickness, hardness, friability, swelling index, differential scanning calorimetry (DSC), particle morphology, in vitro drug release, and cytotoxicity on Ca-Ski cells. Both layers were designed to release both drugs concurrently for a synergistic effect. The polymer-polymer interaction between the layers was able to reduce the loss of formulation due to chitosan. While the bilayer tablet had satisfactory performance in the physicochemical tests, in vitro cell culture with Ca-Ski also showed a synergistic effect using a combination of drugs at a low dose. However, the formulation only had 24-h dose release before degradation. Further drug combinations should be evaluated in subsequent studies.Non-destructive estimation of the internal properties of fruits and vegetables is very important, because better management can be provided for subsequent operations. Researchers and scientists around the world are focusing on non-destructive methods because if they are developed and commercialized, there will be an impressive change in the food industry. In this regard, this paper aims to present a non-destructive method based on Vis-NIR spectral data. https://www.selleckchem.com/products/Everolimus(RAD001).html The different stages of the proposed algorithm are (1) Collection of samples of Gala apples, (2) Spectral data extraction by spectroscopy, (3) Pre-processing of spectral data, (4) Measurement of chemical properties of titratable acidity (TA) and taste index, (5) Selection of key wavelengths using hybrid artificial neural network-firefly algorithm (ANN-FA), (6) Non-destructive estimation of the properties using two methods of hybrid ANN- Particle swarm optimization algorithm and partial least squares regression. For considering the reliability of methods for estimating the chemical properties, the prediction operation was executed in 300 iterations. The results represented that the mean and standard deviation of the correlation coefficient and the root mean square error of hybrid ANN-PSO and PLSR for TA were 0.9095 ± 0.0175, 0.0598 ± 0.0064, 0.834 ± 0.0313 and 0.0761 ± 0.0061 respectively. These values for taste index were 0.918 ± 0.02, 3.2 ± 0.39, 0.836 ± 0.033 and 4.09 ± 0.403, respectively. Therefore, it can be concluded that the hybrid ANN-PSO has a better performance for non-destructive prediction of the two mentioned chemical properties than the PLSR method. In general, the proposed method can predict the chemical properties of TA and taste index non-destructively, which is very useful for mechanized harvesting and management of post-harvest operation.Systemic lupus erythematosus (SLE) is a complex systemic autoimmune disease with heterogeneous clinical manifestations. A diverse innate and adaptive immune dysregulation is involved in the immunopathogenesis of SLE. The dysregulation of immune-related cells may derive from the intricate interactions among genetic, epigenetic, environmental, and immunological factors. Of these contributing factors, non-coding RNAs (ncRNAs), including microRNAs (miRNAs, miRs), and long non-coding RNAs (lncRNAs) play critical roles in the post-transcriptional mRNA expression of cytokines, chemokines, and growth factors, which are essential for immune modulation. In the present review, we emphasize the roles of ncRNA expression in the immune-related cells and cell-free plasma, urine, and tissues contributing to the immunopathogenesis and tissue damage in SLE. In addition, the circular RNAs (circRNA) and their post-translational regulation of protein synthesis in SLE are also briefly described. We wish these critical reviews would be useful in the search for biomarkers/biosignatures and novel therapeutic strategies for SLE patients in the future.SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) infection is associated, alongside with lung infection and respiratory disease, to cardiovascular dysfunction that occurs at any stage of the disease. This includes ischemic heart disease, arrhythmias, and cardiomyopathies. The common pathophysiological link between SARS-CoV-2 infection and the cardiovascular events is represented by coagulation abnormalities and disruption of factors released by endothelial cells, which contribute in maintaining the blood vessels into an anti-thrombotic state. Thus, early alteration of the functionality of endothelial cells, which may be found soon after SARS-CoV-2 infection, seems to represent the major target of a SARS CoV-2 disease state and accounts for the systemic vascular dysfunction that leads to a detrimental effect in terms of hospitalization and death accompanying the disease. In particular, the molecular interaction of SARS-CoV-2 with the ACE2 receptor located in the endothelial cell surface, either at of the disease in high-risk patients.In this article, a Siamese network is applied to the drill wear classification problem. For furniture companies, one of the main problems that occurs during the production process is finding the exact moment when the drill should be replaced. When the drill is not sharp enough, it can result in a poor quality product and therefore generate some financial loss for the company. In various approaches to this problem, usually, three classes are considered green for a drill that is sharp, red for the opposite, and yellow for a tool that is suspected of being worn out, requiring additional evaluation by a human expert. In the above problem, it is especially important that the green and the red classes not be mistaken, since such errors have the highest probability of generating financial loss for the manufacturer. Most of the solutions analysing this problem are too complex, requiring specialized equipment, high financial investment, or both, without guaranteeing that the obtained results will be satisfactory. In the approach presented in this paper, images of drilled holes are used as the training data for the Siamese network.

The aim of the study is to identify the peculiarities of local immune reactions in the skin with underlying soft tissues in patients with different variants of the multiple sclerosis' course. Material and methods The study included 35 patients, hospitalized in the neurological department of the Communal Nonprofit Enterprise of Kharkiv Regional Council «Regional Clinical Hospital» with the established diagnosis of multiple sclerosis. https://www.selleckchem.com/products/17-oh-preg.html The patients were divided into three study groups, based on different variants of this pathology's course. Group 1 included 16 patients with relapsing-remitting type of multiple sclerosis. Group 2 included 11 patients with a secondary-progressive type of multiple sclerosis course. Group 3 included 8 patients with a primary progressive type of multiple sclerosis. Patients of all groups underwent a biopsy of the skin with underlying soft tissues in the lower third of the inner surface of the right lower leg. The comparison group (group 4) was represented by 10 autopsy cases (7 wced violations of local immune reactions in the skin with underlying hypodermis are noted in remitting-relapsing variant of multiple sclerosis course, more pronounced in a secondary-progressing and, especially, primary-progressing variants. Conclusions In patients with multiple sclerosis in the skin with underlying hypodermis activation of T-cell immunity, B-cell immunity and the macrophage system is observed with the development of an immune imbalance between them, characterized by the prevalence of the absolute number of macrophages among all immune cells. Less pronounced violations of local immune reactions in the skin with underlying hypodermis are noted in remitting-relapsing variant of multiple sclerosis course, more pronounced in a secondary-progressing and, especially, primary-progressing variants.The objective of this systematic review was to examine the effectiveness of outdoor exercise park equipment on physical activity levels, physical function, psychosocial outcomes, and quality of life of older adults living in the community and to evaluate the evidence of older adults' use of outdoor exercise park equipment. A search strategy was conducted from seven databases. Nine articles met the inclusion criteria. The study quality results were varied. Meta-analyses were undertaken for two physical performance tests 30-s chair stand test and single-leg stance. The meta-analysis results were not statistically significant. It was not possible to conclude whether exercise parks were effective at improving levels of physical activity. The review shows that older adults value the benefits of health and social interaction from the use of exercise parks. Findings should be interpreted with caution due to the small sample sizes and the limited number of studies.The authors examined if body mass index (BMI), weight, and height across age groups differ between adults with Down syndrome (DS) and adults with intellectual disability but without DS. They conducted secondary analyses of cross-sectional data from 45,803 individuals from the United States from 2009 to 2014 of the National Core Indicators Adult Consumer Survey across five age groups 18-29, 30-39, 40-49, 50-59, and 60+ years. For both men and women with DS, BMI and weight increased between the 18- to 29- and the 30- to 39-year age groups and decreased thereafter. For both men and women with intellectual disability, BMI and weight increased between the 18- to 29- and the 30- to 39-year age groups, stayed about the same until the 50- to 59-year age group, and decreased thereafter. Height demonstrated a small but significant decrease with older age in all groups. These cross-sectional comparisons indicate that BMI and weight may start decreasing at a younger age in adults with DS than in adults with intellectual disability. To investigate the concurrent validity of the International Physical Activity Questionnaire-short form (IPAQ-SF) and the Nordic Physical Activity Questionnaire-short (NPAQ-short) when compared with objectively measured daily steps among older adults. Spearman's ρ between IPAQ-SF and NPAQ-short and objectively measured steps using Garmin Vivofit 3 physical activity monitors. A total of 54 participants were included. The IPAQ-SF subscales' moderate physical activity (PA), moderate to vigorous PA (MVPA), and sedentary time showed little or no correlation with daily steps. The NPAQ-short subscales' vigorous PA, moderate PA, and MVPA showed little or no correlation. The IPAQ-SF subscales' vigorous PA and walking showed fair correlation. Only the IPAQ-SF metabolic equivalent of task minutes showed moderate to good correlation with daily steps. The IPAQ-SF categories and NPAQ-short categorization of World Health Organization compliance were significantly different, but the magnitudes were small and distributions indicated problems with the categorization. The concurrent validity is low, as the scores did not reflect objectively measured daily steps. The concurrent validity is low, as the scores did not reflect objectively measured daily steps.Összefoglaló. Bevezetés és célkitűzés Az obstruktív alvási apnoe (OSA) a felnőtt lakosság jelentős részét érintő betegség, mely ismert rizikófaktora a cardiovascularis és metabolicus betegségeknek és a korai halálozásnak. Mindazonáltal kevés magyarországi adat áll rendelkezésre az OSA-ban szenvedő betegek demográfiai és klinikai jellemzőiről, így vizsgálatunk célja ennek bemutatása volt. Módszer Retrospektív vizsgálatunkban a Semmelweis Egyetem Pulmonológiai Klinikájának Alváslaboratóriumában kivizsgált 394 személy (58 [46-66] év, 73% férfi, testtömegindex [BMI] = 32,5 [29,2-37,5]) adatait elemeztük, akik kitöltötték az Epworth Álmosság Skálát, poliszomnográfiás vizsgálaton estek át; felvettük az anamnézist, és 255 esetben reggeli éhomi vérvétel történt. Eredmények 282 esetben igazolódott OSA. Ebben a csoportban magasabb volt a férfiak aránya (66 vs. 35%), magasabb volt az életkor (59 [48-66] vs. 47 [39-60] év) és a BMI (32,11 [27,78-37,18] vs. 25,29 [22,04-29,03] kg/m2), magasabbak voltak a C-reaktív protein értékek (3,0 [1,71-5,34] mg/l vs.

Using simulations we show that such a converter structure is capable of generating a coherent spin wave beam. By introducing a short wavelength spin wave beam into existing magnonic gate logic, it is conceivable to reduce the size of devices to the micrometer scale. This method gives access to short wavelength spin waves to a broad range of magnonic devices without the need for refined sample preparation techniques. https://www.selleckchem.com/products/epacadostat-incb024360.html The presented toolbox for spin wave manipulation, emission, and conversion is a crucial step for spin wave optics and gate logic.Collective cell migration plays a key role in tissue repair, metastasis, and development. Cellular tension is a vital mechanical regulator during the force-driven cell movements. However, the contribution and mechanism of cell-cell force interaction and energetic costs during cell migration are yet to be understood. Here, we attempted to unfold the mechanism of collective cell movement through quantification of the intercellular tension and energetic costs. The measurement of pN intercellular force is based on a "spring-like" DNA-probe and a molecular tension fluorescence microscopy. During the process of wound healing, the intercellular force along with the cell monolayer mainly originates from actin polymerization, which is strongly related to the cellular energy metabolism level. Intracellular force at different spatial regions of wound and the energetic costs of leader and follower cells were measured. The maximum force and energy consumption are mainly concentrated at the wound edge and dynamically changed along with different stages of wound healing. These results indicated the domination of leader cells other than follower cells during the collective cell migration.In the early 1940s, Paul Flory and John Rehner published a series of papers on the properties of swellable polymeric networks. Originally intended for vulcanized rubber, their development has since been extensively used and extended to much more complex systems, such as hydrogels, and used to estimate the mesh size of such networks. In this article, we take a look at the development of the Flory-Rehner equation and highlight several issues that arise when using such a theory for the described hydrogel networks. We then propose a new approach and equations to accurately calculate the backbone molecular weight in-between crosslinks while explicitly accounting for the molecular mass of the crosslinker and branch segments. The approach also provides more applicable mesh dimensions, for complex networks with macromeric crosslinkers and/or a high degree of branching, as is the case of biocompatible hydrogels. The approach is finally illustrated by a case study comparing the values obtained with our proposed approach to those using the state-of-the-art approach.Luminescence lifetime-based nanosensors for chloride ions were designed by incorporating a luminescent ruthenium dye [Ru(1,10-phenanthroline)3] inside silica nanoparticles and chemically labelling their outer surface with chloride ion-sensitive fluorescent dyes (N,N'-bis(carboxypropyl)-9,9'-biacridine). The nanosensor surface was further functionalized with positively charged amino groups to facilitate intracellular uptake via endocytosis and target lysosomes. The nanosensors have an average diameter of 52 nm and are monodispersed in aqueous solutions. Because of the long lifetime of the reference ruthenium dye, the sensor response can be analyzed using the time-domain dual-lifetime referencing (td-DLR) approach. The use of pulsed excitation in td-DLR rather than intense continuous illumination in ratiometric measurements greatly prevents the dye from photobleaching which significantly improves its measurement stability and reproducibility for long-term monitoring. At optimum conditions, the sensor can measure chloride concentration in the range of 0-200 mM with a large ratiometric signal change from 140.9 to 40.2. Combined with our custom-built microscopic td-DLR system, variations of intracellular chloride concentration in lysosomes were imaged quantitatively with a high spatial resolution and accuracy.Graphene-based e-textile gas sensors have received significant attention as wearable electronic devices for human healthcare and environmental monitoring. Theoretically, more the attached graphene on the devices, better is the gas-sensing performance. However, it has been hampered by poor adhesion between graphene and textile platforms. Meanwhile, amyloid nanofibrils are reputed for their ability to improve adhesion between materials, including between graphene and microorganisms. Despite that fact, there has been no attempt to apply amyloid nanofibrils to fabricate graphene-based e-textiles. By biomimicking the adhesion ability of amyloid nanofibrils, herein, we developed a graphene-amyloid nanofibril hybrid e-textile yarn (RGO/amyloid nanofibril/CY) for the detection of NO2. Compared to traditional e-textile yarn, the RGO/amyloid nanofibril/CY showed better performance in response time, sensing efficiency, sensitivity, and selectivity for NO2. Last, we suggested a practical use of RGO/amyloid nanofibril/CY combined with a light-emitting diode as a wearable e-textile gas sensor.The gut microbiome and its metabolic processes are dynamic systems. Surprisingly, our understanding of gut microbiome dynamics is limited. Here, we report a metaproteomic workflow that involves protein stable isotope probing (protein-SIP) and identification/quantification of partially labeled peptides. We also developed a package, which we call MetaProfiler, that corrects for false identifications and performs phylogenetic and time series analysis for the study of microbiome dynamics. From the stool sample of five mice that were fed with 15N hydrolysate from Ralstonia eutropha, we identified 12 326 nonredundant unlabeled peptides, of which 8256 of their heavy counterparts were quantified. These peptides revealed incorporation profiles over time that were different between and within taxa, as well as between and within clusters of orthologous groups (COGs). Our study helps unravel the complex dynamics of protein synthesis and bacterial dynamics in the mouse microbiome. MetaProfiler and the bioinformatic pipeline are available at https//github.