The Supplemental Nutrition Assistance Program Education (SNAP-Ed), the educational branch of SNAP, can play an important role in improving dietary outcomes, eliminating food insecurity, and preventing chronic disease among low-income populations. This study examined the effects of local SNAP-Ed efforts on self-reported health behaviors and body mass index (BMI) over a 1-year period, using data collected from intercept surveys of program-eligible adults.From 2016 to 2020, the Los Angeles County Department of Public Health partnered with 24 community-based organizations to provide nutrition education and to implement policy, systems, and environmental changes in the community.A cross-sectional survey was conducted in 2018 and repeated in 2019 to measure 6 outcomes describing population-level changes in health behaviors and BMI. The study recruited 4 samples: 2 samples from outside selected supermarkets  and 2 samples from participants at SNAP-Ed class sites .While study results showed an increase in consumption of fruits and vegetables and in vigorous physical activity, they also showed an increase in BMI and high consumption of unhealthy foods. Participating in SNAP-Ed classes was positively associated with several health behaviors but no change in BMI. Participants who experienced food insecurity had worse health behavior outcomes than those who did not experience this condition.SNAP-Ed interventions appear to have a favorable effect on fruit and vegetable consumption, but increases in BMI suggest that unhealthy food consumption is abundant and may be counteracting the benefits gained from eating more fruits and vegetables. Future efforts should take these results into consideration and optimize enrollment in nutrition assistance programs. These efforts should include coordinating with local programs to increase healthy food access for at-risk low-income populations in Los Angeles County. The Supplemental Nutrition Assistance Program Education (SNAP-Ed) can improve dietary and physical activity outcomes. However, despite implementation of SNAP-Ed policy, systems, and environmental changes, the health effects of these interventions are not well understood.Using intercept survey data collected at 2 time points, results showed positive effects of SNAP-Ed programming on several health behaviors but it fell short of eliminating the persistent effect of food insecurity.In addition to increasing access to fresh produce and enrolling eligible populations in nutrition assistance programs, SNAP-Ed implementing agencies should work with partners to address social conditions, such as poverty.Low-income populations are at greater risk for chronic disease because they face a disproportionately higher burden of food insecurity and prevalence of poor dietary consumption ,2. The UWith funding from USDA and state-specific guidance from the California Department of Social Services and the California Department of Public Health, the Los Angeles County Department of Public Health (DPH) operates one of the largest SNAP-Ed programs in the nation. From 2016 to 2020, DPH partnered with 24 implementing agencies to advance policy, systems, and environmental change strategies (PSEs) and to provide nutrition education and promote physical activity in low-income communities in Los Angeles County (LAC). The implementation and evaluation of these efforts are focused on individual-level, direct-interaction nutrition education and physical activity promotion within the community.Participation in a SNAP-Ed nutrition education class has been positively associated with participant nutrition-related self-efficacy, attitudes, and behaviors, such as incorporating fruits and vegetables into meal planning ,5. Some A unique element of SNAP-Ed has been its goal to layer complementary PSEs alongside nutrition education . ExampleOur study sought to address gaps in SNAP-Ed programming by examining data from 2 waves of a cross-sectional intercept survey administered to SNAP-Ed–eligible adults in LAC during 2018 and then again in 2019. The resulting analysis describes SNAP-Ed PSEs and their potential influence on dietary behaviors and body mass index (BMI) for this urban sample.The 2016 to 2020 LAC SNAP-Ed program selected strategies to address diet-related chronic disease risk factors, such as healthy eating and physical activity, at 3 levels of the social-ecological model : individThis local program was developed using the USDA SNAP-Ed Plan Guidance, the USDA Evaluation Framework, and the Dietary Guidelines for Americans –17. DietAn important aspect of the local program was its commitment to partner with community-based agencies that work to improve food quality and food access in LAC. To accomplish this, DPH developed a request for proposals to select 24 organizations that were considered well positioned to reach low-income households throughout under-resourced communities in the county. These agencies were contracted to work within 1 of 8 service planning areas in the county based on their expertise, proposal quality, and population reach.Overall, the evaluation of LAC SNAP-Ed PSEs and programming focused on demonstrating positive changes to USDA priority indicators, including FV consumption, physical activity level, and BMI . We desiFrom 2016 to 2020, the 24 implementing agencies delivered SNAP-Ed nutrition education and implemented PSEs in LAC. These agencies included 17 nonprofit organizations, 2 academic institutions, 3 health care systems, 1 faith-based organization, and 1 school district. All were tasked to carry out 1 required implementation action at each of the 3 levels of the social-ecological model: 1) individual-level nutrition education and physical activity promotion, 2) institutional PSEs, and 3) environmental PSEs. For levels 2 and 3, implementing agencies could choose from a menu of strategies selected by DPH. PSEs specific to LAC were selected and implemented across all 24 partners . This prOver the course of LAC SNAP-Ed, approximately 20,000 direct and indirect nutrition education and physical activity classes and promotional activities were delivered, reaching an estimated 2 million people. Because the SNAP-Ed reporting system did not fully delineate between first-time and repeat attendees, the reach number may contain duplicate program participant entries. To advance the PSEs, most of the funded partners either established new coalitions or joined existing coalitions to coordinate their work in the community. Edible gardens, healthy retail initiatives, and free produce distribution were the most frequently implemented institutional and environmental PSEs. Approximately 300 PSE-related projects were implemented or initiated at some level, reaching approximately 1.2 million people.Our objective was to assess the effects of local SNAP-Ed programming, as indicated by changes in FV servings, SSB consumption, water consumption, energy dense–low nutrient food consumption, days of vigorous physical activity, and BMI. A questionnaire was developed and used to assess these individual-level changes and to estimate the population reach of the overall SNAP-Ed interventions. For the purposes of our study, improvements were defined as changes in the outcomes between 2018 and 2019, time points that account for the varied and broad range of PSEs that were implemented by the 24 implementing agencies.The brief intercept survey collected 2 different adult samples at the 2 time points for a collective total of 4 samples. These samples comprised a general population of adults who were eligible for SNAP-Ed and a group of participants recruited from nutrition education classes that were provided by LAC’s implementing agencies (SNAP-Ed class sites). The intercept method of data collection has been successfully implemented in other studies to recruit low-income and diverse population samples ,21.The general population sample was selected among shoppers at 20 food retail outlets in 2018 and at 15 of the same outlets in 2019. Two large supermarket chains gave permission to contact their customers on the premises. Supermarkets were restricted to neighborhoods where at least 50% of the population in the store census tract was SNAP-Ed–eligible, defined by a household income of less than or equal to 185% of the federal poverty guidelines. SNAP-Ed nutrition education and programs are required to be delivered in these high-need census tracts. All shoppers going in and out of the selected supermarkets were approached and invited to complete a brief questionnaire. To be eligible for the study, participants had to reside in Los Angeles County, be 18 years or older, and speak English or Spanish. Participants or their child(ren) had to be enrolled in 1 of the following programs: SNAP, Special Supplemental Nutrition Program for Women, Infants, and Children (WIC), California Food Assistance Program (CFAP), California Work Opportunity and Responsibility to Kids , General Relief, Child and Adult Care Food Program (CACFP), Head Start, Medicaid , Reduced Lunch, Section 8 housing, Summer Food Program, or Supplemental Security Income (SSI).For the SNAP-Ed class site-specific sample, all program participants present on the day of data collection were asked to participate. Seventeen sites were included in the 2018 sample and 14 of the same sites were included in 2019. The study enrolled adults aged 18 years or older, so that only implementing agencies whose target population was adults participated in the study. Service planning area 5 (West LA) was excluded, as this region did not have any retail food outlets in neighborhoods where at least 50% of residents were SNAP-Ed-eligible.The same questionnaire was administered at all sites in either English or Spanish. The questionnaire was brief, self-administered, and could be completed in 5 to 10 minutes. Questions assessed various health behaviors focusing on those most relevant to the SNAP-Ed mission as well as the interactions between study participants and the implementing agencies. All study participants were offered a $5 gift card.The serial intercept survey used the same questionnaire administered in 2019 as 2018. Data collection occurred during March and April in both years. The only exceptions were that fewer supermarkets and fewer SNAP-Ed class sites were included in 2019, and all study interviewers or data collectors were bilingual in 2019. Five previously participating supermarkets did not renew permission to conduct these surveys, and 3 previously participating SNAP-Ed class sites no longer served adults by 2019 and were not eligible to participate in the second wave of the survey.Six dependent variables were selected, based on the SNAP-Ed Evaluation Framework . The firParticipation in a SNAP-Ed class was captured by asking participants, “Not including today, in the past year, have you taken a nutrition, cooking, or physical activity class sponsored by any of the following Champions for Change groups?” with yes or no response options. Champions for Change was the name for SNAP-Ed in California at the time. For this question, the answer choices were specific to the geographic region where the survey was being administered and listed the name of each implementing agency who delivered services in that service planning area. This was done to help with name recognition and to reduce participant burden by not listing all 24 agencies.The study covariates included age (continuous), sex , and race and ethnicity . Educational attainment categories were less than high school, high school only, and more than high school. Other study covariates were the number of children in the household younger than age 18, whether the participant was a SNAP or CalFresh recipient, Medicaid or Medi-Cal recipient, or WIC recipient, and whether or not they completed the questionnaire in Spanish. Food insecurity was captured by using the 2-question food insecurity tool .Analyses were conducted by using the 2 samples obtained from the supermarkets and the 2 samples obtained from the SNAP-Ed class sites, and study investigators compared responses obtained in 2018 and 2019. Multivariable ordinary least squares regression models were constructed to predict the changes in consumption of FVs, SSBs, energy dense–low nutrient foods, and water; vigorous physical activity (level); and BMI. Outliers with BMI greater than 60 were excluded. The regression analyses controlled for demographic characteristics, public program participation , Spanish language, and food insecurity status. Regression analyses also accounted for the clustering of responses by the supermarket and SNAP-Ed class site samples. All analyses were performed by using Stata version 14 .In the 2018 supermarket sample, 2,098 of the 2,874 (73.0%) shoppers who were approached agreed to participate in the study and completed the intercept questionnaire. In the 2019 supermarket sample, 2,323 of the 3,037 (76.5%) shoppers who were approached agreed to participate and completed the intercept survey. In the 2018 SNAP-Ed class site sample, all 651 program participants (100.0%) who were approached agreed to participate and completed the intercept survey. In the 2019 SNAP-Ed class site sample, 569 of the 634 program participants (89.7%) who were approached agreed to participate and completed the intercept survey.Compared with the 2018 supermarket sample, the 2019 supermarket sample was slightly older. The proportion of women was higher (70.3% vs 54.3%), as was the proportion of participants who were Latino or Hispanic, and the proportion of African Americans was smaller . No signSurvey participants from the SNAP-Ed class site samples were similar in 2018 and in 2019; however, they were older and a higher proportion completed high school in 2019. A smaller proportion of participants were food insecure in 2019 than in 2018 (43.8% vs 57.8%). Across the 2 years, 40.0% of participants had previously taken at least 1 program class.P = .001) (P = .001). Average BMI increased between the years. Among the SNAP-Ed class site samples, a small increase in the average of SSB consumption was observed from 2018 to 2019. Average BMI also increased in these samples, as did average number of days of vigorous exercise .In the supermarket samples, FV consumption increased from an average of 3.3 cups in 2018 to 3.6 cups in 2019 ( = .001) . While tP = .012) and reported 0.34 more days of vigorous physical activity (P < .001) from 2018 to 2019 (P < .001). Among those who participated in a SNAP-Ed class, the increase in FV consumption and vigorous physical activity was of a greater magnitude than that for the other behavior categories. Participating in a class was also associated with an increase in servings of water. Among those who participated in a class, no significant change in BMI was observed. Food insecurity was associated with lower FV consumption, more SSB consumption, more energy dense–low nutrient food consumption, and fewer days of vigorous physical activity.The multivariable regression analyses showed that survey participants from the supermarket samples consumed 0.22 more servings of FV ( to 2019 . ParticiP = .001) yet consumed 0.15 more servings of SSBs (P = .046) from 2018 to 2019 (P = .001). Taking more than 1 SNAP-Ed class, however, was not associated with a significant change in BMI and was not associated with increases in energy dense–low nutrient consumption. Taking more than 1 class was associated with an increase in FV consumption and physical activity. Food insecurity was associated with lower FV consumption, greater SSB, and energy dense–low nutrient food consumption. The multivariable regression analyses from the SNAP-Ed class site samples showed that survey participants reported 0.50 more days of vigorous physical activity  with SNAP-Ed PSEs.SNAP-Ed can play an important role in helping to improve dietary and physical activity outcomes, thereby facilitating opportunities to prevent chronic disease among hard-to-reach, low-income populations. While direct nutrition education is a primary component of SNAP-Ed, delivery of classes can be resource intensive and limited in its scope and reach. Given that millions of people in LAC are eligible for the SNAP-Ed program, scaling complementary PSEs to address structural conditions that can contribute to inequities will be a critical undertaking for the county of Los Angeles, as its health and social services agencies look for cost-effective and sustainable ways to improve health in the community. Future research and evaluation should build on this and other evidence for a better understanding of how SNAP-Ed PSEs affect dietary behaviors and obesity risk from a population health perspective or at the program level.


Comprehensive geriatric assessment (CGA)—a multidimensional diagnostic process to determine medical, cognitive, and functional capacity—has historically included a narrative history supplemented by use of tools to assess domains such as mood or cognition based on assessor preference. This approach to CGA likely works to assess individuals but with increasing clinical complexity and frailty among older adults, a non-standardized approach may mean that key issues are not assessed, and program quality cannot be determined. The COVID-19 pandemic added to these challenges as social distancing practices meant limited face-to-face appointments and use of phone and video assessments. This quality improvement study implemented the interRAI Check-Up Self-Report instrument through a software platform in a specialized geriatric services practice. The instrument can be used over the phone and summarizes specific health problems and needs as well as information about caregiver status and financial trade-offs. Focus groups were also conducted with specialized geriatric services interprofessional team to explore their experiences with implementation. The descriptive analysis of the self-report data revealed expected geriatric issues, such as cognitive and functional impairment, falls and pain. Clients were also commonly experiencing medical instability, cardiorespiratory symptoms, communication impairments, and elevated risk for emergency department visit. Staff found the self-report tool feasible, easy to use, efficient, and the program-level metrics helpful for program planning. In conclusion, introduction of a standardized self-report enhanced CGA by creating a systematic method to flag, track, and prioritize all areas of need for immediate and future care planning at both the client and program level.


The exceptional competitiveness of the orthopedic surgery specialty, combined with the unclear impact of the COVID-19 pandemic on residency recruitment, has presented significant challenges to applicants and residency program directors. With limited in-person opportunities in the 2020-2021 application cycle, applicants have been pressed to gauge chances and best fit by browsing program websites.The aim of the study was to assess the accessibility and content of accredited orthopedic surgery residency program websites during the COVID-19 pandemic.Using the online database of the Electronic Residency Application Service (ERAS), we compiled a list of accredited orthopedic surgery residency programs in the United States. Program websites were evaluated across four domains: program overview, education, research opportunities, and application details. Each website was assessed twice in July 2020, during a period of adjustment to the COVID-19 pandemic, and twice in November 2020, following the October ERAS application deadline.P<.001) and from 0.5% (1/189) to 13.2% , respectively. However, the number of programs that included information about a virtual subinternship or virtual interview on their websites did not change. Over the 4-month period, larger residency programs with 5 or more residents were significantly more likely to add a program video (P<.001) or virtual tour (P<.001) to their websites.A total of 189 accredited orthopedic surgery residency programs were identified through ERAS. Of these programs, 3 (1.6%) did not have functional website links on ERAS. Data analysis of content in each domain revealed that most websites included program details, a description of the didactic curriculum, and sample rotation schedules. Between the two evaluation periods in July and November 2020, the percentage of program websites containing informative videos and virtual tours rose from 12.2% (23/189) to 48.1% (91/189; Most residency program websites offered program details and an overview of educational and research opportunities; however, few addressed the virtual transition of interviews and subinternships during the COVID-19 pandemic. Applicants to orthopedic surgery residencies spend significant time and resources gathering information about potential programs -7. A valDespite the clear utility of websites for residency applicants and programs alike, content is inconsistent and often severely lacking. In a recent review of orthopedic surgery residency websites, Oladeji et al found widespread inconsistencies and noted a scarcity of information desired by prospective applicants . Yong etThe purpose of this study was to assess the content and quality of orthopedic surgery residency program websites and to evaluate adaptations made in response to the COVID-19 pandemic. We hypothesized that, despite the evolving pandemic, updates to program websites, videos, and virtual tours would be limited. This study aims to both describe how orthopedic surgery programs adapted their websites in light of unprecedented circumstances and provide actionable items for programs to improve their online presence during future application cycles.The Electronic Residency Application Service (ERAS) website contains an updated list of all accredited orthopedic surgery residency programs in the United States . Upon acInformation gathered from the orthopedic surgery websites fell into four broad domains: program overview, education, research opportunities, and application details.Program overview included program director name; contact information, including email, phone number, and address; fellowship match lists; wellness opportunities; and salary and benefits information. Efforts to promote diversity were also reported for programs that mentioned underrepresented minorities or gender diversity in their mission statements.The education domain included the mention of extracurricular meetings and courses , didactic sessions, a journal club, sample rotation schedules, clinic and call responsibilities, and educational support, such as funding for loupes and leads.Research opportunities were identified by scanning websites for evidence of a research requirement, publication lists, lab spaces, or funding for national presentations and conferences.Application details gathered from the websites included the number of residents accepted into each program per year, a subinternship description, and guidelines for United States Medical Licensing Examination (USMLE) Step 2 score and Electronic Standardized Letter of Recommendation (eSLOR) submission.Due to the nature of this virtual application cycle, websites were also assessed for the inclusion of program videos, video lengths if applicable, virtual tours, remote opportunities such as virtual subinternships, and details about virtual interviews.t tests, Pearson chi-square tests, and Fisher exact tests. Significance was established at a P value of .05.All data collected from this study were analyzed after the second website review in November 2020. Data from July 2020 were compared to data from November 2020 to assess how programs have modified their websites in response to the virtual application cycle. Unless otherwise noted, statistics were reported on data obtained in July 2020. Analyses were performed using paired-sample Overall, 189 residency programs were identified on ERAS in July 2020. All but 3 residency programs  listed a functional link to the program website. Most programs listed the program director’s name in July 2020  noted didactic sessions, 68.3% (n=129) mentioned a journal club, and 69.8% (n=132) included a sample rotation schedule . Less coMost of the 189 program websites noted a research requirement  and demonstrated research support , such as funding for residents . ResearcThe average number of residents accepted into each program ranged from 4 to 5 residents per year (SD 2.1). Upon reviewing application requirements, only 28.0% (n=53) of 189 websites mentioned a Step 2 score requirement and 14.3% (n=27) requested an eSLOR. P>.99; P<.001), and the percentage of websites including a virtual tour increased from 0.5% (n=1) to 13.2%  and the percentage of virtual tours rose significantly from 0.5% (1/189) to 13.2%  from July to November 2020, the percentage of websites that mentioned a virtual subinternship experience remained stagnant at 6.9% (13/189). This is concerning because, historically, the role of the subinternship in pursuing orthopedic surgery residencies has been to provide both visiting students and programs an opportunity to assess fit based on personal skills, clinical aptitude, and the ability to integrate into program culture [In our study, we accounted for 98.4% (n=186) of the existing 189 orthopedic surgery residency programs through either hyperlinks provided by ERAS or via Google search. We reasoned that the programs would prioritize making COVID-19–related adjustments to websites prior to the ERAS application deadline in October 2020. Therefore, we recorded data in July 2020 and again in November 2020, once the ERAS deadline expired. Our analyses mostly supported our original hypothesis. While the percentage of program videos rose significantly from 12.2% (23/189) to 48.1%  and a virtual tour (P<.001) to their websites between July and November 2020. Further, with less than half  of larger residency programs and less than a quarter  of smaller programs adding a program video during the application season, applicants have been tasked with learning more about orthopedic surgery residency programs using dated videos and online information. Additionally, with only 21% (20/94) of larger programs and 4% (4/95) of smaller programs adding a virtual tour, applicants have limited representations of the physical environment surrounding their potential residency placements. Collectively, these findings indicate that smaller programs were at a potential disadvantage for recruiting applicants since they were less likely than larger programs to have information that applicants would find critical in lieu of in-person interaction.While our results suggest applicants would struggle to find updated information on websites regardless of program characteristics, virtual offerings were also evaluated by program size. Large programs with 5 or more residents were significantly more likely to add a program video  had working websites . In a fo3.4% had For data collection unique to our study, we found that international opportunities were listed on 16.9% (32/189) of websites and fellowship match lists were included on 56.1% (106/189) of websites. While the mention of international opportunities was scarce, the low percentage of fellowship match lists was particularly concerning, given that over 90% of residents choose to complete an orthopedic surgery fellowship following graduation . We alsoPerhaps the most concerning of our findings was the low effort to promote racial and gender diversity on websites. Less than 25% of programs addressed either underrepresented minorities  or gender diversity . Over the past 10 years, racial and gender diversity among orthopedic surgery residency programs has remained stagnant compared to the rise observed in medical schools . DespiteSeveral limitations to this study exist. Since this study started in July 2020, we could not capture COVID-19–related changes prior to this period. Additionally, authors only documented if variables were mentioned on the websites and did not assess the quality of the information. Although data collection by two authors added to the internal validity of the study, we could not control for interrater variability. This study also did not include potential items of interest, such as interview dates, cases performed and their volumes, and operative approaches.The data for this study were collected within a 5-month period from July to November 2020. Although traffic metrics are unavailable, it is reasonable to assume that most applicants visited sites during this time to prepare for ERAS deadlines. This study identified an overall paucity of information on program sites and an inadequate response to the COVID-19 pandemic. Further, websites were difficult to navigate, and important information was dispersed across several tabs. This may have led applicants to overlook time-sensitive application requirements and miss deadlines.We propose several recommendations to improve website quality and quantity of information during the current pandemic and future states of emergency. First, we encourage the ERAS directory of programs to include hyperlinks for all orthopedic surgery residency programs. If a functional hyperlink to a program cannot be found, ERAS should contact the program and encourage it to either provide a link or create a new one if unavailable. Second, all programs should be made aware of standardized information and organization that applicants find useful, such as the ones described in this study. It will ultimately be left to the discretion of the programs whether or not to adjust.Due to the evolving situation of the COVID-19 pandemic, ERAS should encourage and potentially require websites to upload monthly updates. Uploading information about the program’s response to COVID-19 not only has implications for recruitment, but also addresses concerns about safety ,25. SincThis study highlights the inadequate response of orthopedic surgery residency programs to update their websites during this entirely virtual application cycle. As a competitive specialty with the third-lowest specialty match rate, orthopedic surgery programs still have a lot of work to do to improve their online presence, promote diversity, and enhance opportunities for virtual applicants ,19. With


T2 relaxivity (352 Fe mM−1s−1) than a commercial contrast agent Feridex (120 Fe mM−1s−1) at 1.5 T. The alkylation degree of Alkyl-PEI-LAC-TPE effects the restriction of intramolecular rotation process of TPE. Reducing alkane chain grafting ratio aggravated the stack of TPE, increasing the fluorescence lifetime of Alkyl-PEI-LAC-TPE/SPIO nanocomposites. Alkyl-PEI-LAC-TPE/SPIO nanocomposites can effectively labelled HeLa cells and resulted in high fluorescence intensity and excellent MR imaging sensitivity. As an MR/optical imaging probe, Alkyl-PEI-LAC-TPE/SPIO nanocomposites may be used in biomedical imaging for certain applications.Magnetic resonance (MR)/optical dual-mode imaging with high sensitivity and high tissue resolution have attracted many attentions in biomedical applications. To avert aggregation-caused quenching of conventional fluorescence chromophores, an aggregation-induced emission molecule tetraphenylethylene (TPE)-conjugated amphiphilic polyethylenimine (PEI) covered superparamagnetic iron oxide (Alkyl-PEI-LAC-TPE/SPIO nanocomposites) was prepared as an MR/optical dual-mode probe. Alkyl-PEI-LAC-TPE/SPIO nanocomposites exhibited good fluorescence property and presented higher  Recently, aggregation-induced emission (AIE) , 2 has b1T contrast agents have been reported to combine with AIE molecules as a dual-mode imaging probe which realized the liver-specific imaging , 1.31–1.02 [–CH2–2)1010–3CH]. The 1H NMR spectrum of Alkyl-PEI-LAC-TPE with different grafting degrees of alkane chains was displayed in The −1 were attributed to the -NH and at 2940–2820 cm−1 were attributed to the -CH2 of PEI. The characteristic peaks at 3410 cm−1 (-OH), 1650–1560 cm−1 (-C = O) and 1060 cm−1 (-C-O) were attributed to LAC, indicating the successful synthesis of PEI-LAC. For PEI-LAC-TPE, a peak of the stretching vibration of aromatic rings of TPE at 750–700 cm−1 could be seen. The -CH2 peak of Alkyl-PEI-LAC-TPE at 2940–2820 cm−1 was strengthened after modified with alkane chains.FTIR was used to characterize PEI-LAC, PEI-LAC-TPE and Alkyl-PEI-LAC-TPE. As shown in 1H NMR of Alkyl-PEI-LAC-TPE according to the integral values at δ 7.22–6.74 and δ 0.81 ppm  and nitrogen (N) in PEI, PEI-LAC and PEI-LAC-TPE. The results showed that the LAC and TPE grafting ratios were 11% and 10%, respectively. Besides, the alkane chain grafting ratio was calculated by the 0.81 ppm . ResultsOrganic SPIO nanocrystal has a narrow size distribution with a mean diameter of 9 nm characterized by DLS in n-hexane . Alkyl-PAs fluorescence probes, the fluorescence properties of Alkyl-PEI-LAC-TPE micelles and Alkyl-PEI-LAC-TPE/SPIO nanocomposites were studied, respectively. As shown in The fluorescence intensity of gradient alkyl chain Alkyl-PEI-LAC-TPE micelles was measured at a series of TPE concentrations to evaluate the effect of alkane chain grafting ratios on the fluorescence properties of Alkyl-PEI-LAC-TPE. As shown in To understand if the degree of alkane chain grafting ratios has any effect on the fluorescence property of Alkyl-PEI-LAC-TPE micelles, their fluorescence lifetime was measured. Fluorescence lifetime of fluorescence probes can be used to evaluate the impact of microenvironment  around the fluorophores on their fluorescence property . To obtaAs previously reported, restriction of intramolecular rotation (RIR) is the main mechanism for the AIE characteristic of TPE fluorophores , 30, 31.In addition, the fluorescence lifetime of Alkyl-PEI-LAC-TPE/SPIO nanocomposites have also been studied. As shown in T2 relaxivities of Alkyl-PEI-LAC-TPE/SPIO nanocomposites with gradient alkyl chains were measured on a 1.5 T clinical MRI scanner (Siemens). IONP is an effective MR T2 contrast agent, which can significantly shorten the T2 relaxation time of surrounding water protons [T2 relaxivity of the Alkyl-PEI-LAC-TPE (13%)/SPIO, Alkyl-PEI-LAC-TPE (27%)/SPIO and Alkyl-PEI-LAC-TPE (40%)/SPIO nanocomposites was 284.6, 337.2 and 351.8 Fe mM−1s−1, respectively. Results were similar to previous studies that nanocluster containing multiple SPIO nanocrystals performed higher T2 relaxivities than those containing single SPIO nanocrystal [T2 relaxivities of Alkyl-PEI-LAC-TPE/SPIO nanocomposites became higher with the increased alkane chain grafting ratio. One possible reason might be that there are more SPIO nanocrystals stacked in the hydrophobic core of Alkyl-PEI-LAC-TPE/SPIO nanocomposites with higher alkane chain grafting ratios. Moreover, the T2 relaxivities of all Alkyl-PEI-LAC-TPE/SPIO nanocomposites were higher than the commercial contrast agent Feridex  [T2 contrast agent or cell labelling probe. protons , 33. MR ocrystal . What is, 1.5 T) . This imBoth Alkyl-PEI-LAC-TPE micelles and Alkyl-PEI-LAC-TPE/SPIO nanocomposites have shown good fluorescence properties, and their efficacies were tested with cell labelling. Alkyl-PEI-LAC-TPE/SPIO nanocomposites shown good biocompatibility to HeLa cells with TPE concentration at 2.5 µg/ml . CLSM waT2-weighted signal intensity, which was inversely to the number of cells. As shown in T2 value decreased from 216 to 61 ms with the increasing number of Alkyl-PEI-LAC-TPE (13%)/SPIO labelled HeLa cells, which was similar to the results of Alkyl-PEI-LAC-TPE (27%)/SPIO and Alkyl-PEI-LAC-TPE (40%)/SPIO nanocomposites labelled HeLa cells. These nanocomposites had demonstrated excellent MR imaging sensitivity in label HeLa cells and may find potential applications in cell imaging.As a dual-mode MR/fluorescence imaging probe, Alkyl-PEI-LAC-TPE/SPIO nanocomposites should have both MR and fluorescence imaging capabilities. In order to evaluate the probes’ MR visibility, HeLa cells were labelled by Alkyl-PEI-LAC-TPE/SPIO nanocomposites with different Fe concentration  for 12 h. Results  showed tT2 relaxivities, which was higher than that of the commercial MR contrast agent Feridex. The dual-mode imaging probes showed good labelling efficiency for HeLa cells and exhibited excellent MR imaging sensitivity in vitro MRI study under clinical MR scanners. In summary, the Alkyl-PEI-LAC-TPE/SPIO nanocomposites as an IONP and AIE molecular-based MR/optical imaging probe may find appropriate applications in biomedical imaging.In conclusion, we have developed an MR/optical imaging probe by encapsulating multiple hydrophobic SPIO nanocrystals in AIE-based amphiphilic Alkyl-PEI-LAC-TPE polymer micelles. Alkyl-PEI-LAC-TPE/SPIO nanocomposites are spherical with controllable size distribution. These nanocomposites exhibited good AIE properties with bright blue emission. Moreover, lower alkylation degree of Alkyl-PEI-LAC-TPE was helpful for longer fluorescence lifetime and higher fluorescence intensity of Alkyl-PEI-LAC-TPE/SPIO nanocomposites. These nanocomposite probes were efficiently internalized by HeLa cells and mainly accumulated in the cytoplasm with high fluorescence intensity. Besides, Alkyl-PEI-LAC-TPE/SPIO nanocomposites had a high rbab023_Supplementary_DataClick here for additional data file.


The capacitive EIS sensor has been characterized regarding its TMV sensitivity by means of constant-capacitance method. The EIS sensor was able to detect biotinylated TMV particles from a solution with a TMV concentration as low as 0.025 nM. A good correlation between the registered EIS sensor signal and the density of adsorbed TMV particles assessed from scanning electron microscopy images of the SiO2-gate chip surface was observed. Additionally, the isoelectric point of the biotinylated TMV particles was determined via zeta potential measurements and the influence of ionic strength of the measurement solution on the TMV-modified EIS sensor signal has been studied.Plant viruses are major contributors to crop losses and induce high economic costs worldwide. For reliable, on-site and early detection of plant viral diseases, portable biosensors are of great interest. In this study, a field-effect SiO It is widely distributed and infects tobacco, tomato, bell pepper, and other members of the family Solanaceae as well as plants in about 30 further families, while it is harmless for mammals [Harmful organisms, like animal pests, weeds, and plant pathogens, have reduced the productivity of crops since the beginning of agriculture. To avoid such crop losses, farmers have developed various strategies. Nowadays, thanks to modern technologies, crop plants can be protected against a lot of vermin that caused crop loss and famines in the past by using insecticides, fungicides, or genetic modification of the plants . Despiteit crops , the barl Africa , 6 .  mammals . The wor mammals , 11. TMV mammals . During  mammals –15 and b mammals –20.To develop control strategies for plant viral diseases, reliable and early detection methods are required . Enzyme-To overcome these limitations, portable fast, easy-to use biosensor devices for on-site detection are of great interest. As an example for TMV detection, biosensors based on surface plasmon resonance (SPR) and quartz-crystal microbalance (QCM) have been introduced –31. Thes2/Ta2O5 electrolyte-insulator-semiconductor (EIS) structure, where TMV particles loaded on the Ta2O5 surface serve as nano-scaffolds for an extremely dense immobilization of enzymes (penicillinase) [2O5 with a nearly Nernstian pH sensitivity , which represents an important commercial factor for a later large-scale production of on-site, low-cost biosensors.Recently, we reported on a highly sensitive, long-term stable penicillin biosensor based on a nanoparticle-modified, capacitive field-effect Al/p-Si/SiOllinase) . In addillinase) . Here, T58 mV/pH , 43) was58 mV/pH , 45. The38 mV/pH , 47). In2-gate EIS sensors are applied for the electrical detection of charged intact TMV particles loaded from solutions with different TMV concentrations ranging from 0.025 nM (0.001 mg/mL) to 5 nM (0.2 mg/mL), whereby a broader concentration range could be studied than in our previous work [2 surface is evaluated from scanning electron microscopy (SEM) images. For a better understanding of the label-free detection of the TMV particles by their intrinsic charge, zeta potential (ZP) measurements for the determination of the isoelectric point (pI) of the biotinylated TMV particles are conducted for the first time. Finally, an influence of the ionic strength (Debye screening effect) of the measurement solution on the TMV-modified EIS sensor signal is studied and discussed for the first time.The SiOous work . The sen2 layer structure, a p-doped silicon wafer  was thermally oxidized for 30 min at 1000 °C in O2 atmosphere, creating a 30-nm-thick SiO2-gate insulator layer. During this oxidation process, an unwanted SiO2 layer was also formed on the back side of the p-Si, which was removed by etching with hydrofluoric acid. Then, 300 nm Al was deposited on the rear side of the wafer to provide an electrical contact to the Si. In a final step, this wafer was tempered for 10 min in N2 atmosphere at 400 °C. The fabricated wafer was separated to 1 cm × 1 cm chips and cleaned for 5 min in an ultrasonic bath consecutively with acetone, isopropanol, ethanol, and deionized water, respectively.To achieve the Al/p-Si/SiO2PO4  and 10 mM K2HPO4 , pH 7) was stored at 4 °C. For ZP measurements, the TMV stock solution was diluted with 10 mM SPP (15 mM ionic strength) to a concentration of 0.1 mg/mL (2.5 nM) and adjusted to different pH values between pH 7 and pH 3 by titration with 1 M HCl . For the TMV loading experiments, differently concentrated TMV solutions were prepared by diluting the TMV stock solution with 0.1 M SPP buffer, pH 7, and an ionic strength of 0.15 M.As a model plant virus particle, we use a genetically modified TMV variant with a cysteine residue exposed on each coat protein , which wC–V) mode, this leads to a shift of the measurement curve to more positive (or less negative) voltages, which can be seen particularly well in the depletion region of the C–V curve. In the ConCap measurement mode, which is used in this study, the capacitance in the depletion region of the C–V curve  is set constant by applying a sign-inverted voltage to the reference electrode and, thus, compensating potential changes at the gate surface generated due to the adsorption of TMV particles. This way, the EIS sensor response  is recorded dynamically over time. Since the presented field-effect sensor detects TMV particles by their intrinsic charge, the ionic strength of the electrolyte solution represents an important parameter. The charge of the virus particles may partially be screened by counter ions. Thus, the ionic strength of the solution has an impact on the sensor behavior and may also affect the TMV adsorption. Due to that, for all electrolyte solutions used in this study, the concentration as well as the ionic strength is specified.The operation principle of EIS sensors is specified in , and the2 EIS sensors, the EIS chips were mounted in a homemade measurement chamber and sealed with an O-ring, as depicted in Fig. 2 of the SiO2-gate surface was accessible for TMV adsorption and in contact with the electrolyte solution during the electrochemical measurements. The reference electrode  was immersed in the measurement solution and connected to an impedance analyzer . By connecting the Al rear side contact with the impedance analyzer, the electric circuit was closed.For the detection of TMV particles with SiOConCap measurements have been performed in 0.33 mM phosphate-buffered saline (PBS) solution (pH 7) before and after loading the sensor surface with different TMV concentrations; ionic strength was 5 mM. For studying the influence of ionic strength, PBS buffer solutions between 6.5 mM PBS (100 mM ionic strength) and 0.06 mM PBS (1 mM ionic strength) have been used. By diluting a 10-mM PBS stock solution (~154 mM ionic strength) with deionized water, the respective buffer concentrations were achieved. For TMV loading, the sensor chip was incubated with 50 μL of the appropriate TMV solution for 1 h at room temperature in a humid chamber. Afterwards, the sensor was washed three times with 0.33 mM PBS buffer to get rid of non-adsorbed TMV particles.The working principle of the TMV-modified EIS sensor is based on detecting the intrinsic charge of the virus particle. Therefore, the pI  is an important parameter for this detection method. The pI gives information about the sign of a particle charge depending on the pH value. By that, the expected direction of the signal change, induced by the adsorption of charged TMV particles to the gate surface, can be determined. The pI of native TMV  is described to be about pH 3.5: TMV particles are negatively charged at pH values above 3.5 and positively charged at pH values below 3.5 , 51. To 2 surface, SEM images were taken from 6 representative areas of the chip surface, using a JEOL JSM-7800F Schottky field-emission microscope . Before imaging, a ca. 3–5 nm Pt/Pd film was sputtered onto the chip surface. The density of adsorbed TMV particles on the sensor surface was determined from the SEM images using the image evaluation software ImageJ Fiji . For calculating the TMV density, the TMV particle was defined as a 300-nm long nanotube. Though, TMV particles can adsorb as long end-to-end structures, or as smaller particle fractions with lengths often in the range of 50–200 nm. Therefore, the number of the differently sized particles was multiplied with corresponding factors.For morphological characterization of the TMV-loaded SiOFor ZP measurements, the biotinylated TMV particles were dissolved in 10 mM SPP buffer, with a TMV concentration of 2.5 nM. The ZP was measured at different pH values between pH 3 and pH 7. Figure 2-gate surface.As indicated in Fig. 2 surface will alter the gate surface charge. To evaluate the TMV sensitivity of the SiO2 EIS sensor, ConCap measurements have been performed in 0.33 mM PBS buffer (pH 7) before and after loading of virus particles on the sensor surface from differently concentrated TMV solutions. For each TMV concentration, three SiO2-gate EIS chips were characterized. In Fig. Since EIS sensors are charge-sensitive devices and because biotinylated TMV particles are negatively charged at pH > 3.3, it can be expected that loading of TMV particles on the SiOWith increasing TMV concentration from 0.025 to 5 nM, the signal change rises from 22 to 68 mV, evidencing that the charge at the sensor surface becomes more negative as a consequence of an increased number of adsorbed TMV particles. It is worth to note that even at the lowest TMV concentration of 0.025 nM used in this work, the sensor shows a sufficiently detectable sensor signal.2-gate EIS sensors was 13 ± 1.4 mV/dec in the range of 0.025–5 nM. For a TMV concentration of 5 nM, a high standard deviation for the sensor signal can be seen. The reason for this is that the surface density of adsorbed TMV particles on one of the three sensors was significantly lower, which has been supported by scanning electron microscopy examination. Consequently, this particular sensor showed a clearly lower signal amplitude resulting in a high standard deviation. This means that the reproducibility of SiO2 surface treatment procedure and/or TMV adsorption process has to be further improved. At the same time, this underlines that the sensing principle is working properly, since the sensor signal correlates with the number of adsorbed TMV particles on the gate insulator. Concluding, the results displayed in Fig. Figure We have found only a few articles related to the detection of intact TMV particles with biosensors. Dickert et al. reported a linear detection range from 100 ng/mL to 1000 μg/mL by combining molecular imprinting technique and QCM . In the 2-gate surface was studied by SEM. SEM images were taken from the surfaces of the same EIS chips, which were already used for ConCap measurements.Since field-effect EIS sensors detect charge changes on the gate surface induced by the adsorbed TMV particles, the ConCap signal changes in Fig. 9 TMV/cm2 to (4.2 ± 2.4) × 109 TMV/cm2 with increasing the TMV concentration in solution from 0.025 to 5 nM, respectively.In Fig. The bar chart in Fig. e, and is inversely proportional to the ionic strength of the electrolyte solution. Due to the screening of the virus charge by counter ions in the surrounding solution, the EIS sensor signal could be affected significantly by the Debye length/ionic strength and by the distance between the virus charge and the gate surface.It is known that electrolyte-gated field-effect devices, including the capacitive EIS sensors studied in this work, are generally capable to detect potential/charge changes that occur at the gate surface or within the order of the Debye length from the surface , 54–56. In Fig. 2-gate EIS sensors was recorded in PBS buffer (pH 7) with different ionic strengths of 1 mM, 5 mM, 10 mM, and 100 mM . Typical ConCap curves of EIS sensors with TMV particles loaded from the solution with two different TMV concentrations of 0.5 nM and 2.5 nM are depicted in Fig. Therefore, to find out the impact of the charge screening effect, the ConCap signal of TMV-modified SiO2 layers typically have some sensitivity to Na+ ions [+-ion sensitivity for SiO2 of ~2.5 mV/pNa reported in [Figure Na+ ions , 57, theorted in .The results achieved in this experiment underline the significant influence of the ionic strength on the performance of virus-modified EIS biosensors, which should be considered in future studies on the application of EIS sensors for plant pathogen detection.2-gate EIS sensor for the detection of TMV particles as model pathogen is presented. Since the working principle is based on the electric charge of the TMV particles, first, the pI of the utilized biotinylated TMV particles was determined by means of ZP measurements and shown to be in the range of natural virus strains. The capacitive EIS sensors were characterized by ConCap measurements before and after loading of negatively charged TMV particles of different concentrations on the SiO2-gate surface. The mean TMV sensitivity of SiO2-gate EIS sensors was 13 ± 1.4 mV/dec in the concentration range of 0.025–5 nM TMV.Plant viruses are a major cause of crop losses worldwide, leading to enormous economic damage. The best strategy to prevent great crop failures is the early detection of virus infections. Due to numerous advantages, like small size, robustness, fast response time, and compatibility with micro- and nanofabrication technologies, electrolyte-gated field-effect devices are very attractive for on-site, real-time, and label-free detection of plant viruses by their intrinsic charge. In this study, a capacitive field-effect SiO2 surface was additionally studied by means of SEM images, from which the density of TMV particles was evaluated. The calculated mean surface density of adsorbed TMV particles rose from (0.014 ± 0.009) × 109 TMV/cm2 to (4.2 ± 2.4) × 109 TMV/cm2 with increasing the TMV concentration in solution from 0.025 to 5 nM, respectively. Moreover, the ConCap signal shifts correlate well with the density of TMV particles adsorbed on the gate surface: The higher the density of loaded virus particles, the larger was the amplitude of the ConCap signal. In addition, to find out the impact of the charge screening effect, the TMV-modified SiO2-gate EIS sensors were characterized in PBS buffers with different ionic strengths between 1 and 100 mM. As expected, due to the more efficient screening of the TMV charge by counter ions, the amplitude of the ConCap signal of TMV-loaded EIS sensors decreased with increasing the ionic strength. The results achieved in this experiment prove the considerable influence of ionic strength on the performance of virus-modified EIS sensors.The morphology of the TMV-loaded SiOEven though the presented sensor shows a high potential for label-free detection of viral particles, the measurements have only been performed in model analytes so far. In real sample analytes, non-specific adsorption of other charged species may generate false-positive signals or mask the usable signal from the target virus particles of interest. The specificity of EIS sensors and the ability to discriminate the virus type are very important aspects and will be studied in further experiments. As for other kinds of field-effect sensors, specificity can be achieved by means of functionalization of the gate surface with receptor molecules, like antibodies specific to the coat protein of the TMV particle, antibody fragments, aptamers, etc. To reduce/eliminate non-specific adsorption, various strategies such as the use of selectively active blocking agents, pre-purifying the biological liquids, on-chip filtering, separation, desalting, pre-concentration, and other strategies have been discussed in the literature, which, in principle, could also be applicable for EIS sensors. As TMV typically accumulates to high concentrations in infected cells, the prospects for a relatively simple selective enrichment protocol from plant raw material are good, but will need careful comparative evaluation.In conclusion, although the study demonstrates the potential of EIS sensors for label-free detection of intact TMV particles (as a model plant pathogen), this approach could most likely be extended to other plant viruses, too. Capacitive EIS sensors represent an attractive platform for future application as sensors for in-field and real-time, early detection of plant diseases.


We would like to thank the authors for exploring students’ understanding of the National Residency Match Program (NRMP) algorithm,The benefits of mentorship during training are well described.We believe the converse is also true. If a program is ranking an applicant low there may be many reasons, including a strong applicant pool, differences in weightings of the written application and interview, or potentially a poor fit based on the interview. The NRMP algorithm does favor the student; so if all other aspects of that residency program are ideal for the applicant, they may still rank a program highly even after hearing they would be ranked low by the program. Acknowledging that it would be an aberrancy for an applicant to know for certain that they will be ranked low, this information may indicate that the residency program feels the fit is not ideal, and there may be programs perceived as less competitive that might be a better fit and, ultimately, a better match for that applicant.Applicants must assess programs based on many characteristics. Each applicant will determine their own personal algorithm for weighting each of these assessments. Factors that have been important to applicants include location and reputation, but knowing that a program will rank an applicant highly may well shine a light on the “goodness of fit,” which is described as the second most important factor in how applicants rank programs.


T315I-driven chronic myeloid leukemia–like disease and in BCR-ABL1-driven acute lymphoblastic leukemia (ALL). Although crizotinib binds to the ATP-binding site, it also allosterically affected the myristol binding pocket, the binding site of GNF2 and asciminib (former ABL001). Therefore, crizotinib has a seemingly unique double mechanism of action, on the ATP-binding site and on the myristoylation binding pocket. These findings strongly suggest the clinical evaluation of crizotinib for the treatment of advanced and therapy-resistant Ph+ leukemia.Resistance remains the major clinical challenge for the therapy of Philadelphia chromosome–positive (Ph+) leukemia. With the exception of ponatinib, all approved tyrosine kinase inhibitors (TKIs) are unable to inhibit the common “gatekeeper” mutation T315I. Here we investigated the therapeutic potential of crizotinib, a TKI approved for targeting ALK and ROS1 in non-small cell lung cancer patients, which inhibited also the ABL1 kinase in cell-free systems, for the treatment of advanced and therapy-resistant Ph+ leukemia. By inhibiting the BCR-ABL1 kinase, crizotinib efficiently suppressed growth of Ph+ cells without affecting growth of Ph− cells. It was also active in Ph+ patient-derived long-term cultures (PD-LTCs) independently of the responsiveness/resistance to other TKIs. The efficacy of crizotinib was confirmed in vivo in syngeneic mouse models of BCR-ABL1- or BCR-ABL1 Although tyrosine kinase inhibitors (TKIs) directed against BCR-ABL1 have greatly improved the outcome of Philadelphia chromosome–positive (Ph+) leukemia , there iT315I, and all known single BCR-ABL1 mutants in vitro as well as in vivo is approved for treatment of CML and Ph+ ALL patients with multi-TKI-resistant disease or presence of the T315I mutation [Clinically most challenging is the gatekeeper mutation T315I due to its resistance to all approved second-generation TKIs. Only ponatinib, a multi-kinase inhibitor, which targets potently native BCR-ABL1, BCR-ABL1mutation .In advanced Ph+ leukemia, responses to TKIs are limited and transient, despite administration guided by mutational analysis. In these cases, mechanisms of resistance are often unknown but include amplification of BCR-ABL1, aberrant phosphatase activity, drug transporter activity, and “sanctuary sites,” e.g., the CNS .All these therapeutic obstacles including intolerability and adverse side effect emerging with chronic exposition to a given TKI prompted us to explore the therapeutic potential of FDA-approved TKIs with activity against the ABL1 kinase, which revealed crizotinib as one of the most promising candidates. Crizotinib is an inhibitor of mesenchymal-epithelial transition factor (MET)/hepatocyte growth factor receptor (HGFR) and anaplastic lymphoma kinase (ALK) and several other kinases . As crizT315I in Ba/F3 cells showing that it inhibited not only the autophosphorylation of BCR-ABL1 and BCR-ABL1T315I but also the substrate phosphorylation of STAT5  only suppressed BCR-ABL1, but not the “gatekeeper” mutant BCR-ABL1T315I  with different response rates to TKI where a unique culture system allows to culture primary cells without genetic or immunophenotypical changes or any sign of senescence for at least 6 months . PH is  HP Fig. . InteresBCR-ABL1 . The faT315I raised the question of the modalities by which crizotinib inhibited ABL1 kinase activity. It has been shown that crizotinib only binds to the ATP-binding site of ABL1 [The fact that crizotinib exhibited nearly the same effect on native BCR-ABL1 as on BCR-ABL1 of ABL1 . Assumin of ABL1 . This r of ABL1 . The ab of ABL1 . Howeve of ABL1 .T315I in a syngeneic mouse model for CML-like disease [T315i (p = 0.038 for crizotinib and p = 0.014 for ponatinib)  Fig. .Fig. 2TIn order to study the effect of crizotinib in an advanced Ph+ leukemia, we treated syngeneic BCR-ABL1-driven ALL in the same way as CML-like disease with a very similar outcome Fig.  confirmiE255K-T315I to be resistant against ponatinib and PF-114 [E255K-T315I at the same concentration needed for native BCR-ABL1 in contrast to ponatinib which exhibited activity only starting from 125 nM which is far above its clinical feasible concentration.One future clinical challenge upon exposure to third-generation TKIs in high-risk Ph+ leukemia will be the emergence of “compound mutations,” multiple TKD mutations in one BCR-ABL1 allele. Mutation screens predict observations in patients BCR-ABL1d PF-114 , 16. HeIn conclusion, crizotinib appears as a new subclass of inhibitor of BCR-ABL1, binding at the ATP-binding site and, allosterically, affecting the MBP regulatory site. The facts that it (i) is in an advanced stage of clinical development, (ii) acts as an allosteric inhibitor of BCR-ABL1, and (iii) represents a reserve in the case of resistance due to a compound mutation in BCR-ABL1 opens completely new therapeutic options and warrants consideration for clinical testing of crizotinib in Ph+ leukemia patients resistant to available TKI.All compounds  used in this study were purchased from Selleck Chemicals  and were diluted in DMSO to a 1000× stock solution, which then were diluted to 1× working concentrations for the experiments. For the in vivo experiments, the compounds were diluted to working concentration in 0.5% methyl-cellulose .BCR/ABL constructs harboring point mutations related to TKI resistance  were cloned into the retroviral PINCO vector as shown previously [p185eviously .The catalytic domain of His-ABL1 was produced as previously described . In shoAll of the cell lines used in this study were obtained from the German Collection of Microorganisms and Cell Cultures . BV-173 and Jurkat cells were cultured in RPMI 1640 medium supplemented with 10% fetal calf serum (FCS) . SupB15 cells were cultured in RPMI 1640 containing 20% FCS. The packaging cell line Phoenix was cultured in DMSO with 10% FCS. Ba/F3 cells were grown in RPMI with 10% FCS supplemented with 10 ng of murine (m)IL-3 . Ph+ ALL PD-LTCs expressing BCR/ABL  or BCR/ABL-T315I (KÖ) were maintained in serum-free medium as described previously , 19.Western blot analyses were performed according to widely established protocols. The following antibodies were used: anti-ABL , anti-phosphorylated ABat Y245 (α-p-ABL-Y245), anti-STAT5 and anti-phosphorylated STAT5 . Blocking and antibody incubation were performed in 5% low-fat dry milk, followed by washing in Tris-buffered saline  containing 0.1% Tween-20 (TBS-T).50 was calculated using the Erithacus software .Cytotoxicity and proliferation were assessed using the XTT Proliferation Kit , according to the manufacturer’s instructions. Cell growth was assessed by dye exclusion using Trypan blue. Apoptosis was evaluated using 7-amino-actinomycin D (7-AAD) staining, as described previously . The IC+ cells were isolated from the BM of 8–12-week-old female C57BL/6N mice  using the Sca1+ Enrichment Kit according to the manufacturer’s instructions .Sca1+ cells pre-stimulated for 2 days in DMEM, 10% FCS, mIL-3 (20 ng/mL), mIL-6 (20 ng/mL), and mSCF (100 ng/mL) (Cell Concepts) or Ba/F3 cells were plated onto RetroNectin-coated  non-tissue culture 24-well plates and exposed to the retroviral supernatant for 3 h at 37 °C.Ecotropic Phoenix packaging cells were transfected with the PINCO vectors. The retroviral supernatant was collected after 36 h. Sca1All animal studies were conducted in accordance with national animal protection laws and were approved by the relevant monitoring institution (Regierungspräsidium Darmstadt - F 39/08). We used for these experiments a completely randomized design with independent experimental units. We defined cohorts with 7–8 mice/group . The an5 retrovirally transduced donor cells were inoculated via tail vein.For the transduction/transplantation model 8–12-week-old C57BL/6N mice, recipient mice were sub-lethally irradiated with 4.5 Gy. 104) from C57BL/6N mice with BCR/ABL-driven ALL were injected via tail vein into sub-lethally irradiated (4.5 Gy) recipient.For the syngeneic ALL, cryo-preserved BM cells (4 × 102-humidified atmosphere until colonies were visible (2–3 weeks). The plates were stained for 4 h with 5 mg/mL 3--2,5-diphenyltetrazolium bromide (MTT), and the dye was extracted with 1 mL solubilization buffer (20% sodium dodecyl sulfate (SDS), 50% N,N-dimethyl-formamide, 25 mM HCL) for 24 h. The optical density was measured at 570 nm wavelength with a reference wavelength of 630 nm.A colony-forming assay in soft agar was performed as previously described . BrieflThe AlphaScreen assay was performed according to the manufacturer’s general protocol . Reactions were performed in a 25 μL final volume in white 384-well microtiter plates . His-tagged ABL1 (200 nM) and biotinylated myristoylated-peptide derived from the myristoylated N-terminus of ABL1 (myristoyl-GQQPGKVLGDQRRPSLK-Biotin)  (400 nM) were preincubated 30 m in 50 mM Tris-HCl (pH 7.4), 100 mM NaCl, 1 mM dithiothreitol, and 10 μM polyglycine in the presence of 1% DMSO (100% binding) or in the presence of different concentrations of compounds crizotinib or GNF2 diluted in DMSO. Subsequently, 5 μL of beads containing nickel chelate-coated acceptor beads and streptavidin-coated donor beads was added and incubated in the dark for 60′ at room temperature; the emission of light from the acceptor beads was measured in an EnVision reader (Perkin Elmer) and analyzed using the EnVision Spotfire® Analyst software (Perkin Elmer).


How does the current program-centric algorithm for the National Resident Matching Program (NRMP) compare with a student-centric algorithm?In this cross-sectional study of randomized computer-generated data corresponding to the NRMP match for 2018, 2019, and 2020 among more than 50 000 student applicants and 4000 programs in 23 specialties, the 2 algorithms did not differ in percentage of students matched. The student-centric algorithm, relative to the program-centric algorithm, matched a significantly higher percentage of students to their first-ranked program and to their top-5–ranked programs; however, the last position was filled with students who had lower program rankings in the student-centric algorithm vs the program-centric algorithm.These findings suggest that research is needed on these 2 algorithms’ resource demands as well as ensuing resident and program performance. This cross-sectional study of randomized computer-generated data compares program-centric and student-centric algorithms for the National Resident Matching Program. The current program-centric algorithm for the National Resident Matching Program (NRMP) primarily uses the program’s ranking of students to determine a match. Concerns that the existing algorithm favors programs over students, recent findings that the program’s ranking of applicants is not associated with resident performance, and disruptions of existing screening methods and metrics have prompted reevaluation of the current algorithm relative to a student-centric algorithm, in which student ranking of programs is primary and program ranking of students is secondary.To compare program-centric and student-centric algorithms for the NRMP participants.This cross-sectional study used randomized computer-generated data reflecting the NRMP match for 2018, 2019, and 2020, capturing more than 50 000 students and more than 4000 programs in 23 specialties, to compare the 2 algorithms.The same simulated students, programs, and rankings were exposed to the 2 algorithms, running 2300 simulations in the overall analysis and 1000 simulations in each of 23 specialties.The percentage of students who did and did not match, the percentage of students who matched to their top-ranked and top-5–ranked programs, and the program’s rank of the last student matched per position were examined.P = .73). The student-centric algorithm, relative to the program-centric algorithm, matched a significantly higher percentage of students to their first-ranked program  and to their top-5–ranked programs . However, the last position was filled with students who had lower program rankings in the student-centric algorithm vs the program-centric algorithm .The 2 algorithms were not different in percentage of students matched overall (eg, for 2020, program-centric: 59% ; student-centric: 58% ; In this study, the 2 algorithms were not different in the percentage of students matched overall. However, the student-centric algorithm matched a significantly higher percentage of students to their preferred programs. The program-centric algorithm was associated with a lower program’s last matched student rank. Further research is needed on the algorithms’ associations with cost and time demands in the match, postmatch resident and program performance, and fit with a changing environment. The National Resident Matching Program (NRMP) has grown to serve more than 50 000 student applicants to more than 30 000 positions in more than 4000 medical residency programs. Programs invite selected student applicants for interviews. Interviewed students then rank programs by preference and vice versa. Programs and students submit their rank-ordered lists to the NRMP for possible matching through its algorithm.1 Students have to apply, be selected for interviews, schedule interviews, manage and pay for travel, interview, and submit postinterview rankings, in addition to their ongoing academic and clinical work. Similarly, residency program directors and other administrators face a growing number of applicants to be reviewed, invited, interviewed, and rank ordered.2 Concerns and debates have arisen regarding the matching process.3 Surveys of 206 program directors and 314 students participating in the match reveal that many program directors are confused about how the match actually works and feel it is too expensive and time-consuming for the applicants and programs, while most applicants (60%) perceive that the process needs to be changed.4 The NRMP favors programs over students, who are not informed of this bias.5 A counterargument has been that since students received higher-ranked matches than programs, students are not treated unfairly.6 Nonetheless, several groups, including the American Medical Student Association, the Public Citizen’s Health Research Group, and the Medical Student Section of the American Medical Association, argue that the matching algorithm is biased toward residency programs at the expense of students and seek an algorithm that is more favorable to applicants.7 The match process has been legally challenged as uncompetitive, but Congress deemed that the process is ”highly efficient, pro-competitive, and longstanding.”8The process is exhausting for students and programs.9 Some proposals have prompted radical change to residency matching. There has been a call for a modern, free-market approach to residency matching in which hospitals and students negotiate directly year-round.10 Such a proposal would allow competent students to apply and fill positions at any point during the year, which could better suit applicants, programs, and the public. Such a proposal involves substantial changes among many stakeholders, but what are needed are “other more immediate practical solutions to the issues with the Match.”11Several attempts have tried to improve the residency match process for applicants and residency programs.7The matching process in the current program-centric algorithm starts by considering students and programs in order of student preference and then matches to the program are decided by the program’s rankings of students. This algorithm was adopted after heated debate and concerns raised regarding how much the former (pre-1998) matching algorithm was biased toward residency programs at the expense of applicants and interest in whether it would be feasible to replace that algorithm with something more favorable to applicants.7 It has been posited that there would be little difference between a program-optimal algorithm and a student-optimal algorithm and future research should examine the results of the 2 algorithms.12 The existing program-centric algorithm has been used for more than 30 years with little examination or comparison to alternative algorithms.However, the revised program-centric algorithm resulted in few match changes—less than approximately 1 in 1000 students or approximately 16 students total—relative to the previous algorithm.13 The general increase in the number of applications per student over time has not been accompanied by increases in match rates2 and applicant success, while making the application process less personal.14 The residency application process and traditional screening methods have been disrupted by the recent curtailing of educational opportunities and in-person interviews.15 The Step 1 exam, which has been used as screening criterion, is becoming pass/fail. The application process needs to improve for applicants and programs by identifying and using predictive factors of resident success in screening, finding better ways to match applicants with programs, and increasing match rates among female applicants and applicants from underrepresented groups.16 There is a need for outcomes data for medical educators to reevaluate the process and explore changes.17 Incorporating student preferences may be a way of making the recruiting process more personal while also increasing sensitivity to and beginning to meet the expectations of applicants from minority groups. In ranking programs, applicants from underrepresented groups placed a higher weight on patient population, culture, inclusion, and diversity than other students.18 Minoritized applicants seem to restrict their search to programs diverse enough for them to feel comfortable.19 There is a high prevalence of burnout among trainees, which undermines professional development, places patients at risk, and diminishes well-being.20 Permitting a larger role for student preferences in the matching process may improve residents’ fit with program, wellness, trainee experience, and patient care. Incorporating more student preferences may reduce some of the inefficiency in the residency selection process, given that programs and applicants cannot currently discern the true level of interest they have in each other.15New findings have emerged, and the recruiting environment has changed substantially. The NRMP’s program’s rank list has been found of little value in predicting which residents would do best in residency or would take on academic or leadership roles once graduated.This study illustrates the steps in the current program-centric algorithm relative to an alternative student-centric algorithm. It then compares the matching outcomes of the 2 algorithms using simulations of randomized computer-generated students, programs, and positions that reflect NRMP data.Per the Common Rule, this cross-sectional study was exempt from institutional review board or ethics committee review as well as the requirement for informed consent because it did not use actual, private patient, student, or program data.21 is documented and explained in the NRMP’s published video and data.23The The program-centric algorithm begins by considering an individual student’s proposed, prioritized program. The algorithm then matches using the criterion of the program’s ranking of students. Matches are tentative, in that a student may be dematched in a subsequent iteration after considering other students who are ranked higher by the program. The tentative matches are finalized after the iterative steps consider all students and programs.The student-centric algorithm starts by compiling joint ratings, ie, student’s rankings of programs and program’s rankings of the student. The matching criteria are applied at the program level. The algorithm considers a student’s ranking of a program first to possibly match to a program’s positions. If students are tied for the program’s last position(s), the program’s ranking of the students is used to break the tie. The algorithm has fewer steps, and its matches are not tentative.Some observations of the example are as follows. First, the 2 algorithms matched the same number of students . Next, both algorithms filled all the positions of the 2 programs. Third, the 2 algorithms overlapped for most matches . Thus, the 2 algorithms had a common center. Fourth, 2 students (D and A) matched only with the program-centric algorithm, while 2 other students (B and G) matched only with the student-centric algorithm. Fifth, more students matched to their preferred, higher-ranked programs with the student-centric algorithm  than with the program-centric algorithm . Finally, the last student to match with the program had a better rank in the program-centric algorithm (ranks 2 and 4) than with the student-centric algorithm (ranks 4 and 5).19 In the student-centric algorithm, student preferences are given a larger role, but programs do not abdicate all their power. The program would still retain substantial influence through its selection of students for interviews at the beginning of the process, which are needed to submit ranking inputs into the algorithm. At the end of the process, the program would also break any ties using the program’s ranking of students. As more students rank a program highly, the number of ties increases, and the program plays a larger role in breaking ties to determine which of the students match.In the current application system, the student is the least powerful actor.The 2 algorithms were evaluated using the same input data and comparing their matching results. The first step was to generate random data sets based on published NRMP data for 2018, 2019, and 2020. The number of programs and number of students used in each simulation are the same as those published by the NRMP each year and include more than 50 000 students applying to more than 30 000 positions in more than 4000 programs  that follows the steps shown in the The fourth step was to record the match results from the simulations of the 2 algorithms on the same data set. These 4 steps constituted 1 simulation.The matching results were averaged across the simulations for each algorithm. We ran a minimum of 1000 simulations each year to attain a margin of error in the results of less than 3%. The following metrics were measured and compared between the 2 algorithms: the proportions of all students who matched, the proportions of students who matched to their first-ranked program overall and in each of 23 specialties, the proportion of students who matched to their top-5–ranked programs, and the rank of the last student matched to a program per position.12 Accordingly, the null hypotheses were that the 2 algorithms would not be different on these matching outcomes, while the alternative hypotheses were that the 2 algorithms would exhibit differences in these matching outcomes. Analysis was conducted in Excel version 16.49 (Microsoft). Two-tailed tests were performed on these matching outcomes since the student-centric algorithm could possibly have a positive or negative effect. Statistical significance was set at P < .05.It has been posited that a program-optimal and student-optimal algorithm would exhibit little difference.P = .73).The overall match percentages for the 2 algorithms for each of the years 2018, 2019, and 2020 are shown in P < .001; top-5–ranked: 60%  vs 46% ; P < .001). This finding held in each of the 3 years examined.The match percentages of the program-centric and student-centric algorithms to students first-ranked and top-5–ranked programs are also shown in P < .001). This finding held for each of the 3 years examined.The programs’ mean of the ratio of the rank of the last student matched in program to its number of positions appear in 4 Students felt it favors programs over applicants and seek an algorithm more favorable to students.7 The program-centric algorithm matches primarily through the use of the program’s ranking of students, which has recently been found not to be associated with student performance in a residency program, pursuit of academic positions, or leadership achievements.13 These factors prompted a comparison of the program-centric algorithm and a student-centric algorithm, in which student ranking of programs is primary and program ranking of students secondary.The current program-centric algorithm has been used by the NRMP to match students to residency positions for a generation. Program directors have been confused about how the match actually works and feel it is too expensive and time-consuming for both applicants and programs.12 This comparison study found that the 2 algorithms were not different in overall match rates; however, the student-centric algorithm matched more students to their first-ranked and top-5 programs, but it also decreased the program’s ranking of the last matched student compared with the program-centric algorithm.A program-optimal algorithm has been posited to exhibit little difference from an applicant-optimal algorithm.11 If a program’s ranking or student’s ranking improves resident and/or program performance, it provides support for its use in screening applications. Research is also needed to analyze if a combination of these 2 rankings improves performance. Another consideration is how the screening criteria should be adapted to a changing environment. As the average number of applications has increased over time, the process has become impersonal.14 The process may become more personal by incorporating more students’ preferences in the matching algorithm, including those of minoritized students who are attuned to diversity, inclusion, desired patient population, and atmosphere,18 and including preferences of students seeking a personal fit with the program, which may improve resident well-being and patient care.There is a need to examine how the program’s ranking and student’s ranking criteria relate to postmatch resident and program performance. Recent research has found that the program’s ranking of students is not associated with resident performance.This study has limitations. It used randomized computer-generated data on positions, programs, students, and rank-ordered lists based on NRMP data. Future research should include real data on these variables and associate them with matching outcomes as well as with longer-term, diverse indicators of resident and program performance. The matching algorithms relied on a single summary ranking by students and programs. Future research should explore inclusion of multiple rankings and variables, such as geographical affinity, to develop richer matching models. This study showed that the 2 algorithms had relative strengths and weaknesses. Future research should explore how algorithms may be combined to build on their strengths and cover their weaknesses. An algorithm change may have varied intended and unintended consequences that should be researched to guide implementation. Further research is also needed on the relative impacts of the 2 algorithms on the number of applications, interviews, and the cost and time demands on students and programs, which have been increasing over time.In this study, the current program-centric algorithm, which has been used for 30 years without a comparative evaluation, and a student-centric algorithm were similar in overall match rates, but the student-centric algorithm matched more students to their first-ranked and top-5 programs, while the program-centric algorithm increased a program’s rank of the last matched student. The application environment is changing and disrupting traditional application metrics and methods. Research should examine prospects for increasing inputs in algorithms, the effects of these algorithms on resident and program performance after the match, how to adapt them to a changing environment, and whether biases exist in algorithms that differentially affect participants.