Search

El asfalto ecuatoriano tiene la particularidad de ser proclive al envejecimiento prematuro incidiendo en que los pavimentos asfalticos presenten una insuficiente durabilidad. El objetivo de este trabajo experimental es valorar el efecto que produce en las propiedades del asfalto AC 20, procedente de la Refineria de Esmeraldas, su modificacion con el agente rejuvenecedor Sylvaroad RP-1000; asi como evaluar su incidencia en la calidad de una mezcla asfaltica tipica, sin material fresado, preparada con asfalto sin modificar y modificado con rejuvenecedor. El asfalto utilizado clasifica como PG 64-22 y al ser mezclado con 2.5% en peso del rejuvenecedor, su grado cambio a PG 58-28. Se realizaron estudios adicionales empleando los parametros ΔTc, Glover-Rowe y las temperaturas de transicion, evidenciandose mejoras en los resultados. Luego se diseno una mezcla densa con 5.5% de asfalto; a las briquetas confeccionadas se le aplicaron pruebas de modulo de rigidez, compresion ciclica y fatiga con el equipo NAT. Al emplear asfalto modificado el comportamiento a la fatiga mejoro notablemente. Adicionalmente, se aplico un proceso de envejecimiento en horno a 85°C a ambas mezclas, valorandose el modulo y la fatiga a los 8 dias, que demostraron un buen comportamiento solo en la mezcla con rejuvenecedor.

In this chef d’oeuvre, Tomlins offers a heuristic for how to extract the words, ideas, and actions of Nat Turner, the Black, enslaved man who led the most important slave rebellion in American history. Tomlins makes such an effort from within a cluster of different kinds of sources, each one a small window on the past, none of which Turner personally wrote. How to see beyond these particularly distorted glass windows on the past is not obvious. Tomlins’s In the Matter of Nat Turner provides a key not only to Turner, and to his powerful sense of how to fracture the fragile legitimacy of the southern slaveholding elite, but also a metaphysics of interpretive strategy that can serve as a theoretical model.

BACKGROUND Transplantation of kidneys from hepatitis C virus (HCV)-viremic donors into HCV-negative patients followed by direct-acting antiviral therapy was an important breakthrough to increase the number of life-saving kidney transplants. Data suggest that these transplants offer several benefits; however, it is unknown whether adoption of this practice has been shared equitably, especially among disadvantaged groups. METHODS We evaluated United Network for Organ Sharing data on HCV-seronegative adult deceased-donor kidney transplant recipients from January 1, 2017, to June 12, 2020. We compared recipients of a kidney from an HCV antibody- (Ab-)/nucleic acid test- (NAT-), HCV Ab+/NAT-, and HCV NAT+ donor. The primary covariates were as follows: (1) race/ethnicity; (2) female sex; and (3) highest level of education. Models included variables associated with being offered an HCV NAT+ kidney. We fit mixed-effects multinomial logistic regression models with the center as a random effect to account for patient clustering. RESULTS Of 48 255 adult kidney-alone deceased-donor kidney transplant HCV-seronegative recipients, 1641 (3.4%) donors were HCV NAT+-, increasing from 0.3% (January 2017-June 2017) to 6.9% (January 2020-June 2020). In multivariable models, racial/ethnic minorities, women, and those with less education were significantly less likely to receive a kidney from an HCV NAT+ donor relative to an HCV Ab-/NAT- and HCV Ab+/NAT- donor. The disparities were most pronounced among Hispanic and Asian patients with less educational attainment (grade school, high school, or some college/tech school). CONCLUSIONS Despite an increase in transplants from HCV NAT+ donors, we found substantial racial/ethnic disparities in transplantation of these kidneys. These data highlight how the benefits of a scientific breakthrough are often made less available to disadvantaged patients.

Purpose: Increasing tumor-infiltrating lymphocytes (TIL) is associated with higher rates of pathologic complete response (pCR) to neoadjuvant therapy (NAT) in patients with triple-negative breast cancer (TNBC). However, the presence of TILs does not consistently predict pCR, therefore, the current study was undertaken to more fully characterize the immune cell response and its association with pCR. Experimental Design: We obtained pretreatment core-needle biopsies from 105 patients with stage I–III TNBC enrolled in ARTEMIS (NCT02276443) who received NAT from Oct 22, 2015 through July 24, 2018. The tumor-immune microenvironment was comprehensively profiled by performing T-cell receptor (TCR) sequencing, programmed death-ligand 1 (PD-L1) IHC, multiplex immunofluorescence, and RNA sequencing on pretreatment tumor samples. The primary endpoint was pathologic response to NAT. Results: The pCR rate was 40% (42/105). Higher TCR clonality (median = 0.2 vs. 0.1, P = 0.03), PD-L1 positivity (OR: 2.91, P = 0.020), higher CD3+:CD68+ ratio (median = 14.70 vs. 8.20, P = 0.0128), and closer spatial proximity of T cells to tumor cells (median = 19.26 vs. 21.94 μm, P = 0.0169) were associated with pCR. In a multivariable model, closer spatial proximity of T cells to tumor cells and PD-L1 expression enhanced prediction of pCR when considered in conjunction with clinical stage. Conclusions: In patients receiving NAT for TNBC, deep immune profiling through detailed phenotypic characterization and spatial analysis can improve prediction of pCR in patients receiving NAT for TNBC when considered with traditional clinical parameters.

INTRODUCTION: Neoadjuvant therapy (NAT) is an emerging strategy for operable pancreatic ductal adenocarcinoma (PDAC). While neoadjuvant therapy increases multimodal therapy completion, it risks functional decline and treatment drop out. We used decision analysis to determine optimal management of localized PDAC and consider risks faced by elderly patients. METHODS: A Markov cohort decision analysis model evaluated treatment options for a 60-year-old patient with resectable PDAC: 1) upfront pancreaticoduodenectomy or 2) neoadjuvant therapy (NAT). One-way and probabilistic sensitivity analyses were performed. A sub-analysis considered the scenario of a 75-year-old patient. RESULTS: For the base case, NAT offered an incremental survival gain of 4.6 months compared to SF (overall survival: 26.3 vs 21.7 months). In One-way sensitivity analyses, findings were sensitive to recurrence free survival for NAT patients undergoing adjuvant, probability of completing NAT, and probability of being resectable at exploration after NAT. On probabilistic analysis, NAT was favored in a majority of trials (97%) with a median survival benefit of 5.1 months. In altering the base case for the 75-year-old scenario, NAT had a survival benefit of 3.8 months. CONCLUSIONS: This analysis demonstrates a significant benefit to neoadjuvant therapy in patients with localized PDAC. This benefit persists even in the elderly cohort.

A XXI. századi kompetenciák (IKT kompetencia, média- és információs műveltség, hatékony tanulási készség) egyre nagyobb hatással vannak az életünkre. Mind a magán, mind a szakmai életünkben használjuk az egyes készségeket, képességeket, kompetenciákat, amelyeket folyamatosan tanulnunk, fejlesztenünk kell. Pár évtizede senki nem gondolta volna, hogy a digitális vagy az innovációs kompetenciát kulcskompetenciaként fogjuk megjeleníteni. Pedig így lett, és egyre nagyobb teret hódítanak az egyes eszközök, technológiák, szoftverek alkalmazásával összefüggő készségek, emellett pedig kitüntetett figyelem jut a tanulási képességeknek, az élethosszig tartó tanulásnak, hiszen mindig képesnek kell lennünk új ismeretek befogadására, készségek alkalmazására. Tanulmányunkban a 2020-as Nemzeti Alaptantervet vizsgáljuk meg szöveg- és tartalomelemzés segítségével olyan aspektusból, hogyan jelennek meg benne a XXI. századi kompetenciák, elsősorban az információs műveltség és a digitális kompetencia, illetve ezek részkészségei. Vizsgálatunk alapján elmondható, hogy a 2020-as NAT elemeiben tartalmazza az egyes XXI. századi kompetenciákat, azonban nem egyenlő mértékben, némelyik csak érintőlegesen kap szerepet benne, az információs műveltség pedig fogalmi szinten sem jelenik meg a dokumentumban. Pozitív jelenségként értelmezhető, hogy a kompetenciafejlesztésen belül a digitális források használata mellett a hagyományos dokumentumok és ezek használatának képessége is szerephez jut. Ám az egyes tantárgyakon belül elvárt készségszintek sajnos több helyen nem épülnek egymásra, így inkoherens módon történik a kompetenciafejlesztés.

Background Nucleic acid amplification testing (NAT) for blood screening has been previously performed in some countries to determine NAT yields. The current study sought to explore the non-discriminating reactive NAT yields using individual-NAT (ID-NAT) and characteristics of HBV NAT yields through a 10-year retrospective analysis in Zhejiang, China. Methods Blood donations were analyzed using individual-NAT mode by the transcription-mediated amplification (TMA) method. Supplementary HBV serological tests were performed using chemiluminescent immunoassay, and HBV viral load assay was performed by real-time polymerase chain reaction. Follow-up studies were performed in partial donors with low HBV viral loads. Results Non-discriminating reactive NAT yields and HBV NAT yields varied in different years. The yields ranged from 853.73 per million to 2018.68 per million and 624.60 per million to 1669.50 per million, respectively. In the 476 NAT yields, 19 were probable window periods (WP), 33 probable occult hepatitis B virus infections (OBIs), 409 were confirmed OBIs and 15 were chronic HBV infections. ID-NAT results were categorized in four groups, and the findings showed that the levels of HBV DNA viral loads were different in the four different groups (χ2 = 275.02, p 2 = 49.429, p Conclusion NAT screening of blood donations can reduce the risk of transfusion-transmitted HBV infections. Positive proportions of anti-HBs and anti-HBc are correlated with the HBV viral load level. However, low level of viral load donors pose risks in HBV NAT assays, and show fluctuating state for HBV viral load and leads to non-repeated NAT results during follow up studies.

In animal tissues, N-acyltransferase (NAT) catalyzes the first reaction in the biosynthetic pathway of bioactive N-acylethanolamines, in which an acyl chain is transferred from the sn-1 position of the donor phospholipid, such as phosphatidylcholine, to the amino group of phosphatidylethanolamine, resulting in the formation of N-acylphosphatidylethanolamine. NAT has long been known to be stimulated by Ca(2+), and hence it has been referred to as Ca(2+)-dependent NAT. On the other hand, members of the phospholipase A/acyltransferase (PLA/AT) family (also known as HRAS-like suppressor family) show Ca(2+)-independent NAT activity. In this chapter, we describe (1) partial purification of Ca(2+)-dependent NAT from rat brain, (2) purification of recombinant PLA/AT-2, and (3) NAT assay using radiolabeled substrate.

Je voudrais tout d’abord remercier l’administrateur et la professeure Edith Heard, qui – au nom des biologistes – a présenté le projet d’une chaire annuelle Biodiversité et écosystèmes à l’Assemblée des professeurs, puis ma candidature à cette première chaire, mais aussi la fondation Jean-François et Marie-Laure de Clermont-Tonnerre, qui a apporté son soutien financier généreux, ainsi que ma famille, bien sûr, mes amis et mes collègues. Cette chaire est la dernière en date d’une longue tradit...

International journal of molecular sciences 23(18), 10805 - (2022). doi:10.3390/ijms231810805, Most eukaryotic proteins are N-terminally acetylated by a set of N$\alpha$ acetyltransferases (NATs). This ancient and ubiquitous modification plays a fundamental role in protein homeostasis, while mutations are linked to human diseases and phenotypic defects. In particular, Naa50 features species-specific differences, as it is inactive in yeast but active in higher eukaryotes. Together with NatA, it engages in NatE complex formation for cotranslational acetylation. Here, we report Naa50 homologs from the filamentous fungi $Chaetomium$ $thermophilum$ and $Neurospora$ $crassa$ with significant N- and C-terminal extensions to the conserved GNAT domain. Structural and biochemical analyses show that CtNaa50 shares the GNAT structure and substrate specificity with other homologs. However, in contrast to previously analyzed Naa50 proteins, it does not form NatE. The elongated N-terminus increases Naa50 thermostability and binds to dynein light chain protein 1, while our data suggest that conserved positive patches in the C-terminus allow for ribosome binding independent of NatA. Our study provides new insights into the many facets of Naa50 and highlights the diversification of NATs during evolution., Published by Molecular Diversity Preservation International, Basel

Voice transmission over the Internet network is now taken for granted. Many end-user applications address this issue. However, this paper focuses on the specific use of the SCCP protocol created by Cisco, its implementation in a computer network and end devices, determination of the operational properties of this implementation, and their comparison in different conditions. VoIP traffic is compared at different bandwidths and implemented by different configurations of IP protocols. By investigated implementations of IP protocols are meant IPv4, IPv6, and IPv4 protocol with applied NAT. As part of the application of various IP protocols is also compared VoIP communication with a video stream on a local basis. The conclusion of the paper is devoted to the graphical evaluation of these observations and to draw conclusions based on them.

Efficient transcription termination relying on intrinsic terminators is critical to maintain cell fitness by avoiding unwanted read-through in bacteria. Natural intrinsic terminator (NIT) typically appears in mRNA as a hairpin followed by approximately eight conserved uridines (U-tract) at the 3' terminus. Owing to their simple structure, small size, and protein independence, assorted NITs have been redesigned as robust tools to construct gene circuits. However, most NITs exert functions to adapt to their physiological requirements rather than the demand for building synthetic gene circuits, rendering uncertain working performance when they are constructed intact in synthetic gene circuits. Here, rather than modifying NITs, we established a data-driven and in silico-assisted (DISA) design framework to forward engineer minimal intrinsic terminators (MITs). By comprehensively analyzing 75 natural intrinsic terminators from Bacillus subtilis, we revealed that two pivotal features, the length of the U-tract and the thermodynamics of the terminator hairpin, were involved in the sequence-activity relationship (SAR) of termination efficiency (TE). As per the SAR, we leveraged DISA to fabricate an array of MITs composed of in silico-assisted designed minimal hairpins and fixed U-tracts. Most of these MITs exhibited high TE in diverse Gram-positive and Gram-negative bacteria. In contrast, the TEs of the NITs were highly varied in different hosts. Moreover, TEs of MITs were flexibly tuned over a wide range by modulating the length of the U-tract. Overall, these results demonstrate an efficient framework to forward design functional and broad host-range terminators independent of tedious and iterative screening of mutagenesis libraries of natural terminators.

Objective: The objective of the study is to compare three techniques, routinely used rapid diagnostic tests (lateral flow immune chromatography) versus nucleic acid amplification test (NAT) versus Paper-based microfluidics for DNA diagnostics of Malaria, in terms of their sensitivity and specificity as diagnostic tests in detecting malarial infection among febrile illnesses, suspected of malaria, as well as to compare their cost-effectiveness. Methodology: Three seventy febrile cases suspected of malaria with negative results with RDT will be screened by real-time PCR and DNA microfluidics techniques, sensitivity and specificity of these as screening tests will be compared. The number of extra positive cases detected by NAT gives us the yield. Cost-effectiveness analysis will be done by calculating the incremental cost-effectiveness ratio (ICER) and average cost-effectiveness ratio (ACER) for the tests. Statistical Analysis: Statistical analysis will be done using SPSS version 21. Sensitivity, specificity, Positive predictive values will be computed. Comparison of sensitivity and specificity of NAT, a paper microfluidic technique for DNA diagnostics and RDT will be carried out using McNemar’s test. Receiver operating curves will be generated separately to assess the utility of the NAT. Conclusion: The Implications of this study from the patient's perspective would mean early diagnosis which forms the tenet of control of the disease by increasing the yield. Early diagnosis at the community level would translate into the application of efficient prevention mechanisms to spread the infection. The cost-effectiveness analysis would provide a scientific basis for the adoption of the best test for the diagnosis, given the economic feasibility of the study.

Despite its increased application in pancreatic ductal adenocarcinoma (PDAC), complete response to neoadjuvant therapy (NAT) is rare. Given the critical role of host immunity in regulating cancer, we sought to correlate baseline inflammatory profiles to significant response to NAT. PDAC patients receiving NAT were classified as responders (R) or nonresponders (NR) by carbohydrate antigen 19-9 response, pathologic tumor size, and lymph node status in the resected specimen. Baseline (treatment-naive) plasma was analyzed to determine levels of 27 inflammatory mediators. Logistic regression was used to correlate individual mediators with response. Network analysis and Pearson correlation maps were derived to determine baseline inflammatory mediator profiles. Forty patients (20R and 20NR) met study criteria. The R showed significantly higher overall survival (59.4 vs. 21.25 mo, P=0.002) and disease-free survival (50.97 vs. 10.60 mo, P=0.005), compared with NR. soluble interleukin-2 receptor alpha was a significant predictor of no response to NAT (P=0.045). Analysis of inflammatory profiles using the Pearson heat map analysis followed by network analysis depicted increased inflammatory network complexity in NR compared with R (1.69 vs. 1), signifying a more robust baseline inflammatory status of NR. A panel of inflammatory mediators identified by logistic regression and Fischer score analysis was used to create a potential decision tree to predict NAT response. We demonstrate that baseline inflammatory profiles are associated with response to NAT in PDAC, and that an upregulated inflammatory status is associated with a poor response to NAT. Further analysis into the role of inflammatory mediators as predictors of chemotherapy response is warranted.

The role of lifestyle behaviors on neck adipose tissue (NAT), a fat depot that appears to be involved in the pathogenesis of different cardiometabolic diseases and in inflammatory status, is unknown. In this cross-sectional and exploratory study, the authors examined the relationship between sedentary time and physical activity (PA) with neck adiposity in young adults. A total of 134 subjects (69% women, 23 ± 2 years) were enrolled. The time spent in sedentary behavior and PA of different intensity were objectively measured for 7 consecutive days (24 hr/day), using a wrist (nondominant)-worn accelerometer. The NAT volume was assessed using computed tomography, and the compartmental (subcutaneous, intermuscular, and perivertebral) and total NAT volumes were determined at the level of vertebra C5. Anthropometric indicators and body composition (by dual-energy X-ray absorptiometry) were determined. The time spent in light physical activity and moderate physical activity (MPA) and the overall PA were inversely associated with the intermuscular NAT volume in men, as were the MPA and overall PA with total NAT volume (all ps ≤ .04). Sedentary time was directly related to the total NAT volume (p = .04). An opposite trend was observed in women, finding a direct relationship of MPA with the subcutaneous NAT; of light physical activity, MPA, and overall PA with the perivertebral NAT; and of light physical activity with total NAT volumes (all ps ≤ .05). The observed associations were weak, and after adjusting for multiplicity, the results became nonsignificant (p > .05). These findings suggest that the specific characteristics of PA (time and intensity) might have sex-dependent implications in the accumulation of NAT.

The gold standard for HIV screening of blood donors is individual nucleic acid amplification testing (NAT). However, individual NAT testing is cost-prohibitive, especially in a resource-limited setting. The fourth-generation ELISA that detects both p24 antigen and antibody to HIV-1 and 2 has been recommended as the minimum test for HIV to enhance blood transfusion safety and can be an alternative to NAT testing in resource-limited settings. The aim was to assess the performance of a fourth-generation ELISA in use at a regional blood transfusion service using nucleic acid amplification testing on units of screened blood negative to HIV. The study was a cross-sectional study conducted at the National Blood Transfusion Service center and the Plateau State Virology Research Centre, both in Jos, Nigeria. Between August and October 2016, one thousand and eighteen voluntary blood donors were recruited consecutively and had their samples tested using fourth-generation ELISA. One thousand p24 antigen-negative samples were pooled for NAT in an aliquot of 50 samples. All the pools of fifty samples of 1,000 HIV p24 antigen-antibody negative donor blood screened by the fourth-generation ELISA tested negative for HIV RNA on nucleic acid amplification. The yield of pooled NAT for HIV after a fourth-generation ELISA screening of blood donors was found to be zero in this study, thus establishing the fourth-generation ELISA's reliability. Therefore, we recommend adopting the fourth-generation ELISA test as a minimum requirement for blood donor screening.

To compare overall survival (OS) in patients who underwent surgery for early-stage pancreatic adenocarcinoma (rPca) based on sequence (NAT, neoadjuvant therapy and/or AT, adjuvant therapy) and type (SA, single-agent or MA, multi-agent) of chemotherapy received.Using the National Cancer Database, patients with clinical stage I/II rPca diagnosed between 2010 and 2014 were identified and five comparison matches (1: NAT vs. upfront resection (UR); 2: multi-agent neoadjuvant (MA NAT) vs. single-agent adjuvant therapy (SA AT), single-agent neoadjuvant therapy (SA NAT), multi-agent adjuvant therapy (MA AT); 3: MA NAT vs. MA AT; 4: NAT + AT vs NAT; 5: NAT + AT vs AT) were constructed using minimum distance matching strategy. Median OS (mOS) was analysed using Kaplan-Meier method, log-rank test and Cox proportional hazard model.A total of 18,470 patients with stage I/II rPca were eligible for analysis. NAT showed a 5 month (mo.) improved OS compared with UR (3271 patients/group, 28.1 vs 23.2 mo. P 0.0001 hazard ratio [HR]: 0.79). MA-NAT was shown to be superior to other chemotherapy approaches SA AT, SA NAT, and MA AT (1349 patients/group: 30 vs. 25.9 mo., P = 0.0001 [HR: 0.82]). MA NAT showed a survival advantage over MA-AT (1349 patients/group, 30 vs 26.1 mo., P = 0.0008 [HR: 0.86]). The combination of NAT and AT showed a better outcome when compared with NAT alone (1128 patients/group, 31.6 vs 27.4 mo., P = 0.0011 [HR: 0.81]) or AT alone (1128 patients/group, 31.6 vs. 25.2 mo., P 0.0001 [HR: 0.76]).In patients with stage I/II rPca, MA NAT showed improved mOS compared to UR and all other chemotherapy sequences except both NAT plus AT. These findings support the use of MA NAT in stage I/II rPca patients and warrant prospective trials evaluating MA NAT and post-resection maintenance therapies.

Long noncoding RNAs or lncRNAs are a class of non-protein-coding RNAs that are >200 nt in length. Almost 50% of lncRNAs during zebrafish development are transcribed in an antisense direction to a protein-coding gene. However, the role of these natural antisense transcripts (NATs) during development remains enigmatic. To understand NATs in early vertebrate development, we took a computational biology approach and analyzed existing as well as novel data sets. Our analysis indicates that zebrafish NATs can be divided into two major classes based on their coexpression patterns with respect to the overlapping protein-coding genes. Group 1 NATs have characteristics similar to maternally deposited RNAs in that their levels decrease as development progresses. Group 1 NAT levels are negatively correlated with that of overlapping sense-strand protein-coding genes. Conversely, Group 2 NATs are coexpressed with overlapping protein-coding genes. In contrast to Group 1, which is enriched in genes involved in developmental pathways, Group 2 protein-coding genes are enriched in housekeeping functions. Group 1 NATs also show larger overlap and higher complementarity with the sense-strand mRNAs compared to other NATs. In addition, our transcriptomics data, quantifying RNA levels from cytoplasmic and nuclear compartments, indicates that Group 1 NATs are more abundant in the cytosol. Based on their expression pattern, cytosolic nature, and their higher complementarity to the overlapping developmental mRNAs, we speculate that Group 1 NATs function post-transcriptionally to silence spurious expression of developmental genes.

Background Testing plays a critical role in treatment and prevention responses to the COVID-19 pandemic. Compared to nucleic acid tests (NATs), antigen-detection rapid diagnostic tests (Ag-RDTs) can be more accessible, but typically have lower sensitivity and specificity. By quantifying these trade-offs, we aimed to inform decisions about when an Ag-RDT would offer greater public health value than reliance on NAT. Methods Following an expert consultation, we selected two use cases for analysis: rapid identification of people with COVID-19 amongst patients admitted with respiratory symptoms in a ‘hospital’ setting and early identification and isolation of people with mildly symptomatic COVID-19 in a ‘community’ setting. Using decision analysis, we evaluated the health system cost and health impact (deaths averted and infectious days isolated) of an Ag-RDT-led strategy, compared to a strategy based on NAT and clinical judgement. We adopted a broad range of values for ‘contextual’ parameters relevant to a range of settings, including the availability of NAT and the performance of clinical judgement. We performed a multivariate sensitivity analysis to all of these parameters. Results In a hospital setting, an Ag-RDT-led strategy would avert more deaths than a NAT-based strategy, and at lower cost per death averted, when the sensitivity of clinical judgement is less than 90%, and when NAT results are available in time to inform clinical decision-making for less than 85% of patients. The use of an Ag-RDT is robustly supported in community settings, where it would avert more transmission at lower cost than relying on NAT alone, under a wide range of assumptions. Conclusions Despite their imperfect sensitivity and specificity, Ag-RDTs have the potential to be simultaneously more impactful, and have a lower cost per death and infectious person-days averted, than current approaches to COVID-19 diagnostic testing.

The ongoing pandemic of coronavirus disease 2019 threatens the whole world, which catalyzes a variety of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid test (NAT) kits. To monitor test quality and evaluate NAT kits, quality control materials that best simulate real clinical samples are needed. In this study, the performance of SARS-CoV-2 cell culture supernatant, PCDH-based pseudovirus, and MS2-based pseudovirus as quality control materials was compared. PCDH-based pseudovirus was found to be more similar in characteristics to SARS-CoV-2 particle, and more suitable for evaluating SARS-CoV-2 NAT kits than MS2-based pseudovirus. Proper detection using sensitive and precise NAT kits is essential to guarantee diagnosis. Thus, limit of detection, precision, anti-inference ability, and cross-reactivity of NAT kits from PerkinElmer, Beijing Wantai Biological Pharmacy Enterprise Co, Ltd, Shanghai Kehua Bio-Engineering Co, Ltd, Sansure Biotech Inc., Da An Gene Co, Ltd, Shanghai BioGerm Medical Biotechnology Co, Ltd, and Applied Biological Technologies Co, Ltd, were compared using PCDH-based pseudovirus. For the seven kits evaluated, N gene was more sensitive than ORF1ab gene in most kits, whereas E gene was most sensitive among the three genes in Shanghai Kehua Bio-Engineering Co, Ltd, and Applied Biological Technologies Co, Ltd. PerkinElmer got the lowest limit of detection for N gene at 11.61 copies/mL, and the value was 34.66 copies/mL for ORF1ab gene. All of the kits showed good precision, with CV values less than 5%, as well as acceptable anti-interference ability of 2 mg/L human genomic DNA. No cross-reactivity was observed with other respiratory viruses.

The total synthesis of (+)-03219A, a rare Δ8,9-pregnene isolated from the marine-derived Streptomyces sp. SCSIO 03219, is described that is based on a series of transformations that enable progression from epichlorohydrin to an ent-estrane, then conversion to a nat-androstane, and finally establishment of the natural product target. Key to the success of these studies was implementation of two rearrangement processes to formally invert the quaternary center at C13 and establish the C10 quaternary center.

Background: Treatment patterns in hormone receptor (HR)+/human epidermal growth factor receptor 2 (HER2)− eBC continue to evolve. While AT remains the standard of care, NAT has been indicated as a viable approach to downstage higher-stage tumors. As interest in NAT trials increases, detailed data on current treatment patterns may inform future research. The objective of this retrospective observational study was to evaluate patient characteristics and treatment patterns in US patients diagnosed with HR+/HER2− eBC. Methods: This analysis included adults diagnosed with non-metastatic (stage I-III) invasive eBC between January 1, 2011, and March 1, 2019, in the nationwide Flatiron Health electronic health record–derived de-identified database. All patients had HR+/HER2− test results within 90 days of eBC diagnosis and evidence of both primary surgery and systemic treatment in the eBC setting. Patients with records of HER2-targeted treatment were excluded. NAT was defined as systemic treatment before first primary surgery; AT was defined as systemic treatment within 12 months after primary surgery among patients with ≥ 6 months of follow-up after surgery. Pathologic complete response (pCR) was defined as the absence of residual invasive disease in the breast and axillary lymph nodes after NAT and was subsequently confirmed at the time of primary surgery. Results: Among 4739 patients in the study, 341 (7%) received NAT (318/341 received AT following NAT) and 4398 (93%) received AT only. The proportion of patients receiving NAT increased annually from 5% in 2011 to 9% in 2018. Of NAT-treated patients, 7% achieved pCR. Patients who had higher pathology stage, higher tumor grade, younger age and who were Black or Hispanic/Latino had higher proportions of NAT (Table). Patients who received NAT were more likely to receive a mastectomy (vs lumpectomy) compared with patients who received AT only. The most common NAT regimens included chemotherapy (54%; primarily anthracycline + cyclophosphamide + taxane [ACT] regimens) or endocrine (ET) monotherapy (36%; primarily aromatase inhibitor [AI]). The most common AT regimens were ET monotherapy (79%; primarily AI or tamoxifen). Conclusions: Although AT remains the standard treatment approach in US clinical practice for HR+/HER2− eBC, interest in NAT has grown in recent years and NAT use has increased. Given the low pCR rates following NAT and since pCR is considered a surrogate marker for survival, further investigation into longer-term clinical benefits of NAT in this population is warranted as NAT may be becoming more common. Table. Characteristics of patients with HR+/HER2− eBC who received AT or NATAT only(first treatment after surgery)N = 4398NAT(first treatment before surgery)N = 341pCR achievedYesNA25 (7.3)NoNA271 (79)UnknownNA45 (13)Age at eBC diagnosisMedian (IQR), years64 (54-72)59 (49-68)RaceNon-Hispanic White2975 (68)208 (61)Non-Hispanic Black278 (6.3)31 (9.1)Non-Hispanic Asian120 (2.7)13 (3.8)Non-Hispanic other428 (9.7)27 (7.9)Hispanic/Latino282 (6.4)39 (11)Unknown315 (7.2)23 (6.7)Year of eBC diagnosis2011/20121023 (23)53 (16)2013/20141136 (26)81 (24)2015/20161129 (26)101 (30)2017/2018/201911100 (25)106 (31)Pathology group stageI2370 (54)70 (21)II1209 (27)86 (25)III282 (6.4)58 (17)Unknown537 (12)127 (37)Tumor grade11268 (29)41 (12)22297 (52)186 (55)3768 (17)111 (33)Unknown65 (1.5)3 (0.9)HistologyIDC3486 (79)267 (78)ILC615 (14)51 (15)Other/Unknown297 (6.8)23 (6.7)Primary surgery typeLumpectomy2911 (66)128 (38)Mastectomy1487 (34)213 (62)Data are n (%) unless otherwise specified.AT, adjuvant chemotherapy; eBC, early breast cancer; IDC, invasive ductal carcinoma; ILC, invasive lobular carcinoma; NA, not applicable, NAT, neoadjuvant chemotherapy; pCR, pathologic complete response.* The number of patients diagnosed in 2019 is very low because we only included patients diagnosed through March 2019; thus 2019 is grouped with 2017/18. Citation Format: Mary K Downer, Christopher Craggs, Preet Dhillon, Patricia Luhn, Carlos Flores-Avile, Amie Tan, Monika Patre, Frauke Schimmoller, Tanja Badovinac Crnjevic. Neoadjuvant and adjuvant therapy (NAT/AT) treatment patterns and clinical characteristics in a real-world cohort of US patients with HR+/HER2− early breast cancer (eBC) [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS7-23.

The early determination of response to neoadjuvant therapy (NAT) in triple-negative breast cancer would enable the treating oncologist to adapt the therapeutic regimen of a non-responding patient (e.g., by changing dosage, dose schedule, prescribed drugs), and thereby improve treatment outcomes while avoiding unnecessary toxicities. To address this challenge, we propose to use personalized, in silico forecasts of tumor response to therapeutic regimens via a mechanistic mathematical model calibrated with patient-specific longitudinal multi-parametric magnetic resonance imaging (MRI) data acquired early in the course of NAT. Here, we extend our mechanistic model to include a new term describing the synergistic effects of NAT drug combinations and identify the driving parameters involved in its formulation by means of a sensitivity analysis. Our model describes tumor cell dynamics as a combination of proliferation, which is regulated by a logistic term, and mobility, which is described as a diffusion process constrained by the local tumor-induced mechanical stress. Tumor cell density is extracted from diffusion-weighted MRI data, while tissue mechanical properties are defined from segmented T1-weighted MRI data. We adjust the tumor proliferation rate in response to NAT drug combinations with a recent model of drug synergy, MuSyC, which accounts for distinct types of synergistic drug effects (synergy of potency vs. synergy of efficacy). We also consider the heterogeneous intratumoral delivery of drugs by means of perfusion maps estimated from dynamic contrast-enhanced MRI data. We use Sobol’s method for the sensitivity analysis of two different tumors - one well-perfused and one poorly-perfused. We simulate a four-cycle NAT protocol in which NAT drugs are delivered every 14 days, and assess the total effect (ST) of each parameter on the mean relative difference of tumor cell density with respect to a control simulation of tumor growth without NAT. Sensitivity analysis results directly depend on the definition of the parameter space, which we construct by combining two approaches. First, we experimentally constrain parameter ranges using time-resolved, high-throughput, automated microscopy assays to capture the changes in proliferation rates of various breast cancer lines (HCC1143, SUM149, MDAMB231, and MDAMB468) caused by two standard drug combinations: paclitaxel with carboplatin and doxorubicin with perfosfamide (metabolic derivative of the pro-drug cyclophosphamide), and fitting the MuSyC model to these data. Second, we scale the resulting in vitro parameter ranges to clinically-relevant in vivo ranges by running an in silico study with our mechanistic model of breast cancer growth and NAT response. Our results show that, out of the ten parameters involved in the synergy term, three have a dominant role in the dynamics of breast cancer during NAT (ST > 0.1): synergistic potency, the maximal change in tumor cell proliferation by the slowest decaying drug, and its concentration producing half of maximal effects. The other parameters have marginal (0.02 < ST < 0.1) to negligible effect (ST < 0.02). Ongoing studies are assessing the ability of our mechanistic model to forecast NAT response over a small patient cohort after patient-specific calibration of the driving parameters identified in the present study. Citation Format: Guillermo Lorenzo, Angela M. Jarrett, Christian T. Meyer, Darren R. Tyson, Vito Quaranta, Thomas E. Yankeelov. Identifying relevant parameters that characterize the early response to NAT in breast cancer patients using a novel personalized mechanistic model integrating in vitro and in vivo imaging data [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS13-44.

Aim: Δ9-Tetrahydrocannabinol (Δ9-THC) is a potentially addictive cannabinoid. Its impact on the activity of liver arylamine N-Acetyltransferase (NAT) has not been reported. This study investigated the rewarding effects of Δ9-THC in mice and whether Δ9-THC had any impact ex-vivo and in-vitro on NAT activity. Methods: Thirty-six Swiss albinomice randomly assigned to six groups (n = 6) completed a biased, 8-week Conditioned Place Preference (CPP) paradigm. Mice exhibiting ~80% preference for the black chamber at pre-conditioning were selected. Treatment groups were administered Δ9-THC (0.10, 0.50 or 2.0 mg/kg/mL, ip) or amphetamine (AMP, 5.0 mg/kg/mL, ip); while untreated groups (controls) received vehicle solutions (coconut oil or 0.9% saline). Entries and time spent in the white, drug-paired chamber during a 15-min post-conditioning exploration of the CPP apparatus were compared with the pre-conditioning exploratory scores. Livers from Δ9-THC treated and untreated mice were excised and NAT enzyme activity determined ex-vivo using a spectrophotometric assay with p-anisidine as substrate. The impact of varying concentrations of Δ9-THC (0.00 - 162 μM) on the activities of NAT from untreated mice livers were also investigated in-vitro. Results: Δ9-THC treated mice entered and spent significantly more time in the drug-paired CPP chamber (p ≤ 0.05) at post-conditioning vs pre-conditioning (F = 11.22). Mice treated with 2.0 mg/kg Δ9-THC made significantly more entries into the drug-paired chamber (p ≤ 0.05) as compared with their vehicle controls. AMP-treated mice displayed significant (p In-vitro NAT evaluations revealed a dose-dependent inhibitory impact of Δ9-THC on NAT activity with an IC50 value of 34.97 ±1.58 μM. Ex-vivo evaluations of livers from Δ9-THC treated mice showed no significant impact on liver NAT enzyme activity. Conclusion: Δ9-THC induced significant conditioned place preference (drug reward) and produced a moderate dose-dependent inhibition on NAT activity in-vitro, but not ex-vivo.

BACKGROUND: With the improvement of the efficacy of neoadjuvant therapy (NAT) that is guided by molecular subtypes, the rate of pathologically node-negative disease after NAT (ypN0) is increasing for HER2 positive (HER2+) and triple-negative (TN) breast cancer patients. The necessity of axillary surgery for patients with high ypN0 has been questioned. This study aimed to identify patients among HER2+ and TN breast cancer with low risk for axillary metastases after NAT, and, perhaps, they are suitable for selective elimination of axillary surgery staging. METHODS: From January 2010 to August 2018, 865 breast cancer patients who underwent NAT were included in this retrospective clinical study, and 184 patients (21.3%,184/865) suffered from TN and HER2+ breast cancer and received full-course NAT. The correlation among clinicopathological characteristics of HER2+ and TN breast cancer and ypN0 were analyzed. RESULTS: Among the 184 HER2+ and TN breast cancer patients, tumor staging, lymph node staging and Ki-67 before NAT, clinically node-negative disease after NAT (ycN0), and breast radiologic and pathologic complete response (bpCR) were correlated with ypN0 (P

Digital PCR has shown great potential for quantitative nucleic acid testing (NAT), but most existing platforms are dependent on large auxiliary equipment (e.g., vacuum pump, amplification instrument, fluorescence microscope) to achieve target dispersion, amplification, signal capture and result analysis. Such complex, expensive and bulky NAT platforms have limited their applications in resource-limited areas, especially for point-of-care testing (POCT). In this work, we designed a digital isothermal NAT platform based on a pump-free open droplet array microfluidic chip. A pump-free microfluidic chip was developed based on an open microdroplet array in the form of thousands of independent microdroplets for spontaneous sample dispersion, without the need for external power. Combined with a handheld fluorescent signal reader based on a smartphone, this digital NAT platform can accurately quantify as low as 1 copy per μL of λDNA. Therefore, our integrated NAT platform, as a potable, robust and low-cost tool for highly accurate NA quantitative analysis, holds great potential for POCT applications.

In Italy, the West Nile Virus surveillance plan considers a multidisciplinary approach to identify the presence of the virus in the environment (entomological, ornithological, and equine surveillance) and to determine the risk of infections through potentially infected donors (blood and organ donors). The costs associated with the surveillance program for the Lombardy Region between 2014 and 2018 were estimated. The costs of the program were compared with a scenario in which the program was not implemented, requiring individual blood donation nucleic acid amplification tests (NAT) to detect the presence of WNV in human samples throughout the seasonal period of vector presence. Considering the five-year period, the application of the environmental/veterinary surveillance program allowed a reduction in costs incurred in the Lombardy Region of 7.7 million EUR. An integrated surveillance system, including birds, mosquito vectors, and dead-end hosts such as horses and humans, can prevent viral transmission to the human population, as well as anticipate the detection of WNV using NAT in blood and organ donors. The surveillance program within a One Health context has given the possibility to both document the expansion of the endemic area of WNV in northern Italy and avoid most of the NAT-related costs.