One of the important pathogeneses of eustachian tube dysfunction (ETD) is nasal inflammatory disease. The prevalence of allergic rhinitis (AR) in adults ranges from 10 to 30% worldwide. However, research on the status of eustachian tubes in AR patients is still very limited.This prospective controlled cross-sectional study recruited 59 volunteers and 59 patients with AR from Sun Yat-sen Memorial Hospital. Visual analogue scale (VAS) scores for AR symptoms and seven-item Eustachian Tube Dysfunction Questionnaire (ETDQ-7) scores were collected for both groups. Nasal endoscopy, tympanography and eustachian tube pressure measurement  were used for objective assessment. All AR patients underwent 1 month of treatment with mometasone furoate nasal spray and oral loratadine. Then, the nasal condition and eustachian tube status were again evaluated.p < 0.0001). Nasal endoscopic scores were also positively correlated with ETDQ-7 scores, with a correlation coefficient of 0.7328 (p < 0.0001). After 1 month of treatment, VAS scores of nasal symptoms, endoscopic scores and ETDQ-7 scores were significantly decreased in AR patients (p < 0.0001), and TMM examination also suggested that eustachian tube function was significantly improved after treatment (p < 0.0001).TMM examination revealed that 22 patients  among the AR patients and 5 healthy controls  had abnormal eustachian pressure. Twenty-two AR patients (37.3%) and 9 healthy controls had an ETDQ-7 score ≥ 15. With regard to nasal symptoms of AR, the VAS scores of nasal obstruction were correlated with the ETDQ-7 scores, and the correlation coefficient was r = 0.5124  Registered 12 January 2020—Retrospectively registered, http://www.chictr.org.cn/edit.aspx?pid=48328&htm=4 Eustachian tubes maintain middle ear ventilation and pressure balance, enable mucus clearance and protect the middle ear from pathogens and sounds. Eustachian tube dysfunction (ETD) is estimated to affect 1% of adults and 40% of children . ProlongAR affects more than 500 million people worldwide . In ChinThe aim of this study was to comprehensively assess eustachian tube function in patients with AR and to determine the main nasal symptoms and local factors that contribute most to ETD.Dermatophagoides pteronyssinus (Der p1). The exclusion criteria were as follows: (1) a history of CRS or rhinitis; (2) a history of chronic otitis media (COM) or otitis media with effusion (OME); (3) a history of acute upper respiratory tract infection (AURT) in the last 4 weeks and (4) a history of glucocorticoid or antihistamine use within the last 4 weeks. Healthy volunteers aged 18–60 were collected at the same time for control group. The inclusion criteria for control group included (1) without any nasal or ear symptoms; (2) no abnormal nose and ear signs were found by endoscopy and (3) SPT excluded allergic diseases. The exclusion criteria were as follows: (1) a history of CRS, rhinitis; (2) a history of COM or OME; (3) a history of AURT in the last 4 weeks and (4) a history of glucocorticoid or antihistamine use within the last 4 weeks. We eventually included 59 AR patients and 59 healthy controls. More details for the definition of AR could be found in Additional file This study was approved by the Sun Yat-sen Memorial Hospital Institutional Review Board. We recruited consecutive adult AR patients aged 18–60) who visited Sun Yat-sen Memorial Hospital from June 2019 to January 2020 due to their nasal symptoms. The AR subjects represented the outpatients of ENT clinic in a tertiary hospital. AR was diagnosed according to the criteria of the Initiative on Allergic Rhinitis and its Impact on Asthma (ARIA) . The inc8–60 who All patients were asked to complete a visual analogue scale (VAS) assessment for nasal symptoms, because the VAS score was significantly related to the rhinoconjunctivitis quality of life questionnaire (RQLQ) score and was considered a simple and effective method for assessing AR . The sevhttp://www.graphpad.com). Descriptive analyses were utilized to describe the sample characteristics, and the data are presented as number (%), mean ± standard deviation (SD), or median  according to the tests of normality. The Chi square tests for categorical variables and the paired or unpaired Student’s t tests or Wilcoxon rank sum tests for continuous variables were performed. Spearman or Pearson correlation coefficient analyses were applied to assess the correlation between VAS scores and ETDQ-7 scores, depending on the results of the normality test. Chi square tests were used to compare the categorical data. A p value < 0.05 was considered to indicate significance. Considering that the same patient has two ears, our statistical analyses for Tympanometry and TMM tests were performed on the number of ears.Statistical analysis was performed with GraphPad Prism  and 59 volunteers  were included in the study. The clinical characteristics of the two groups were shown in Table 2 = 7.394, p = 0.0065)  had an ETDQ-7 score ≥ 15, significantly more than the 9 patients (15.3%) in the control group . Twenty-two AR patients  and 6 control subjects  had abnormal TMM results, and the difference between groups was statistically significant   than in the control group  (χp < 0.0001). However, other symptoms did not show significant correlations with ETDQ-7 scores. The Pearson correlation coefficients  were as follows: itching, r = 0.0707 (p = 0.5947); sneezing, r = 0.2203 (p = 0.0937); and rhinorrhoea, r = −0.03006 (p = 0.8212). The Pearson correlation coefficient  between ETDQ-7 and nasal endoscopic scores was r = 0.7328 (p < 0.0001), suggesting a strong correlation . A moderate correlation was found between ETDQ-7 scores and nasal obstruction scores . Three mild AR patients  and 19 moderate/severe AR patients  had abnormal TMM results, and the difference between groups was statistically significant . With regard to subjective symptoms, significantly more moderate/severe AR patients  than mild AR patients  had ETDQ-7 scores ≥ 15  (Table The patients with AR were classified into a mild group (22 patients) and a moderate/severe group (37 patients). We found that there were significantly fewer type A tympanograms in the moderate/severe AR group  than in the mild AR group  . The prevalence of abnormal TMM was decreased significantly to 10 patients  . The symptoms of ETD were also improved significantly. We found that only 8 AR patients (13.6%) had ETDQ-7 scores ≥ 15 after treatment, significantly fewer than the 22 patients (37.3%) before treatment   and nasal endoscopic scores (p = 0.0006) were significantly higher in the uncontrolled-ETD group. This suggests that uncontrolled AR symptoms, especially nasal obstruction, might contribute to irreversible ETD  and patients with ETDQ scores < 15 (the controlled-ETD group). We found that after 1 month of treatment, the nasal obstruction VAS scores (ETD Fig. .Fig. 4AfETD is a well-recognized comorbid manifestation of inflammatory diseases of the nasal cavity and nasal sinus . HoweverNasal VAS scores and ETDQ-7 scores were used to assess symptoms of the nose and eustachian tubes in our study. According to the subjective symptom scores, approximately 37.3% of AR patients had ETD, while up to 51.4% of moderate/severe AR patients had ETD. Among the four major symptoms of AR, nasal obstruction showed a moderate correlation with ETDQ-7 score. This means that more severe AR or a greater degree of nasal obstruction is more likely to cause ETD. Many patients with severe AR are prone to concomitant inferior turbinate hypertrophy  and toruObjective measures were also included in our study. Tympanograms, TMM test results and nasal endoscopic scores were taken for every AR patient. According to the TMM test results, 33.1% of AR patients had ETD, and 44.6% had severe AR combined with ETD. Nasal endoscopic scores for the pharyngeal orifices of eustachian tubes exhibited a strong correlation with ETDQ-7 scores. This suggests that the local conditions of the pharyngeal orifices of the eustachian tubes are closely related to ETD. Moreover, we found that the same patients could have different nasal endoscopic scores on the left and right sides. This could also explain why some AR patients only had unilateral ETD.Our study also revealed that treatment with loratadine and mometasone furoate effectively improved nasal symptoms and significantly decreased ETDQ-7 scores and the frequency of abnormal TMM test results in AR patients. Both objective and subjective scores showed good consistency before and after treatment. Intranasal corticosteroids and antihistamines are recommended for treating perennial AR . AlthougTo our knowledge, this is the first pilot study on ETD in AR and to find that AR can cause ETD. However, there are some limitations. First, our study involved a referral system and a single centre; thus, the ETD cases may not reflect the epidemiology of ETD in all AR populations. Second, this study was not based on a double-blinded, randomized controlled trial. Third, although certain characteristics, such as smoking, laryngopharyngeal reflux and atopy, are not known to be risk factors for ETD, these factors were not considered in our analysis. Then, other nasal symptoms like itching, sneezing and rhinorrhoea showed very weak correlations with ETDQ-7; this may be caused by insufficient sample size. Finally, we used only oral antihistamines and nasal mometasone furoate for AR patients with ETD; other treatment approaches that can help to improve eustachian tube function, such as Buteyko breathing and Valsalva training, were not included in our study.In conclusion, AR patients, especially those with severe nasal obstruction, are more likely to have ETD than individuals without AR. ETD in patients with AR is mainly characterized by ear blockage and ear pressure, and the appearance of symptoms is usually associated with the onset of rhinitis, which should attract attention in clinical work. Both ETDQ-7 scoring and TMM are effective means of assessing eustachian tube function in AR patients. Regular treatment for AR can reduce patients’ nasal symptoms and effectively improve eustachian tube function.Additional file 1. Table S1. The degree of skin prick test. Table S2. The degree of specific immunoglobulin E level in serum. Figure S1. Specific immunoglobulin E levels to the 8 common allergens in AR patients. Figure S2. Treatment results of nasal symptom, ETDQ-7 and endoscopic scores in AR patients compared with control group. Table S3. The treatment results of eustachian tube dysfunction for AR patients.


Background: We analyzed mRNA profiles of prostate cancer related genes in circulating tumor cells (CTCs) of primary, non-metastatic triple-negative breast cancer (TNBC) patients (pts) before and after neoadjuvant chemotherapy to elucidate the potential of prostate cancer targets in this BC subgroup.Method: Blood from 41 TNBC pts (n = 41 before / 26 after therapy) was analyzed for CTCs applying the AdnaTest EMT-2/Stem Cell Select. Multimarker RT-qPCR allowed the detection of the prostate specific antigen PSA, the prostate specific membrane antigen PSMA, full-length androgen receptor (AR-FL), and AR splice-variant seven (AR-V7).Results: Before therapy, at least one prostate cancer related gene was detected in 15/41 pts (37%). Notably, in 73% of AR-FL positive cases, AR-V7 was co-expressed. After therapy, CTCs of only one patient harbored prostate cancer related genes. AR-V7+ and PSMA+ CTCs significantly correlated with early relapse  whereas PSMA+ CTCs also associated with a reduced OS (p = 0.0059). This correlation was confirmed for PSMA+ CTCs in univariate , but not multivariate analysis.Conclusion: Although CTCs that expressed prostate cancer related genes were eliminated by the given therapy, PSMA+ CTCs significantly identified pts at high risk for relapse. Furthermore, AR inhibition, often discussed for this BC subgroup, might not be successful in the primary setting of the disease since we identified AR-FL+ CTCs together with AR-V7+ CTCs, associated with therapeutic failure. Triple-negative breast cancer (TNBC), accounting for 15–20% of all breast cancers (BC), has an destructive behavior which is associated with poor prognosis , 4 and cLooking for new predictive biomarkers, prostate cancer related markers have been evaluated in TNBC for additional treatment options. In this context, based on findings in prostate cancer (PCA), the prostate specific membrane antigen (PSMA) has become an attractive molecular target for oncological imaging and radionuclide therapy using PSMA PET/CT in TNBC , 11. In Comparable with data for HR, concordance of AR expression status between primary BC tissue and metastatic lesions was shown to be 15–35% . ConsequAR which was detected in 33% of the patients before but rarely after therapy (AR-FL) wildtype itself but AR splice variants, and in particular AR variant seven (AR-V7), have been linked to resistance toward anti-AR drugs like enzalutamide and abiraterone  in blood as a minimal invasive approach to assess the real time AR status –33. Most therapy . Howeverraterone . In thisV7+ CTCs , 37. In reatment .AR-FL, AR-V7, PSA, and PSMA in blood samples of 41 TNBC patients before and 26 TNBC patients after therapy to elucidate their prognostic value and their potential as therapeutic targets.Based on the current findings, the complex interplay of AR and the prostate specific antigen (PSA) and PSMA  and the The clinical characteristics of all patients evaluated before and after therapy are shown in via EpCAM, HER2, and EGFR, a patient was defined as CTC+ if overexpression of one of the four prostate cancer related genes was detected. Before therapy, at least one prostate cancer related gene was detected in 15/41 pts (37%). The expression of AR-FL was documented in 11/41 patients (27%), AR-V7 in 8/41 patients (20%), PSMA in 6/41 patients (15%), and PSA in 5/41 patients (12%), respectively. Notably, as apparent from AR-FL+ cases before therapy, AR-V7 was co-expressed. In 26 of the in total 41 patients analyzed before therapy, we were able to perform CTC analysis also after therapy. In only one of these 26 patients after therapy, we found CTCs with an overexpression of prostate cancer related genes . Using immunomagnetic selection PSMA+ CTCs were more often found in patients experiencing no pCR, as compared to those achieving one. Although these findings were not significant . 8/41 (20%) of the patients died, eight of them BC specific, after a median survival time of 25 months (range: 3–38 months).PSMA+ CTCs . PSMA+ CTCs (p = 0.0059) while AR-V7+ CTCs (p = 0.051). While half of the pts showing PSMA+ CTCs relapsed within 19 months after first diagnosis, more than half of the pts with PSMA- CTCs did not experience a relapse within the period of follow-up  and OS (p = 0.015), respectively. Using multivariate Cox proportional hazard analysis, neither PSMA+ CTCs nor AR-V7+ CTCs, independently associated with a significant shorter PFS  as well as lymph node involvement before therapy (p = 0.028) and non-pCR (p = 0.01) as unfavorable variables for PFS which was confirmed in multivariate analysis for non-pCR (p = 0.041) and lymph nodes before therapy (p = 0.008). For OS, univariate analysis identified tumor size before (p = 0.034), after therapy (p = 0.041) and non-pCR (p = 0.041) to associate with shorter OS.rter PFS  nor withrter PFS . With reERCC1+ CTCs, associated with resistance, as an unfavorable factor for PFS (p = 0.026) and OS (p = 0.017)  .p = 0.01 and p = 0.02). Consequently, comprehensive analysis of CTCs could probably direct physicians to stratify TNBC patients for additional treatment options. The same holds true for prostate cancer related genes, especially AR, which has frequently been discussed to be a target for treatment of TNBC patients.TNBC remains a subtype with a very aggressive behavior and worse outcome , 2. ThusPSMA+ CTCs before therapy significantly identified patients with worse outcome. In the context of AR inhibition, often discussed for the TNBC subgroup, this therapeutic approach might not be successful in the primary setting since we detected AR-FL+ CTCs together with AR-V7+ CTCs, associated with therapeutic failure.We here demonstrated that prostate cancer related genes expressed on CTCs in primary, non-metastatic TNBC patients were mainly found before but rarely after therapy, thus, were eliminated by the given therapy. However, vice versa with an overall disconcordance of 58%  in CTCs from PCA patients  transcripts from as low as a single AR-V7+/K19+ cell , diagnosed between January 2013 and August 2018, were enrolled. All patients presented with first diagnosis of TNBC in our clinic, were non-metastatic and had not been treated before. Blood was obtained after written informed consent from all subjects using protocols approved by the clinical ethic committee of the University Hospital Essen (05/2856). Patient characteristics are documented in The study was conducted at the Department of Gynecology and Obstetrics, at the University Hospital of Essen, in Germany. In total, 41 early TNBC patients (before therapy: n = 39) or adjuvant treatment (n = 2) ; 4 = no tumor detectable. pCR was defined as regression 4 according to Sinn, no evidence of residual invasive cancer and DCIS, both, in breast and axilla; pathological partial response (pPR) was defined as regression 1–3 according to Sinn . Samples were stored at 4°C and processed not later than 4 h after blood withdrawal.2 x 5 ml EDTA blood were collected for CTC isolation in S-Monovettes25-beads and was reverse transcribed . The final reaction volume of 40 μl cDNA was stored at −20°C.Positive immunomagnetic selection targeting EpCAM, EGFR, and HER2  was employed for CTC isolation from 2 × 5 ml blood. The method has been described in detail elsewhere . mRNA waProstateCancerPanel AR-V7 detecting CD45 (PTPRC), PSA (KLK3), PSMA (FOLH1), full-length AR (AR-FL), AR splice variant seven (AR-V7), and GAPDH  has been described in detail recently  with 18 PCR cycles. PCR was performed as follows: denaturation for 5 min at 95°C, followed by 18 cycles of 30 s at 95°C, 90 s at 60°C, and 90 s at 72°C. Preamplified cDNA  was analyzed in duplicates for one of the six transcripts in a reaction volume with miRCURY SYBR Green MasterMix  and ROX Reference Dye  of in total 15 μl. RT-qPCR was performed with the StepOnePlus™  real-time system as follows: PCR activation for 10 min at 95°C, followed by 35 cycles 10 s at 95°C, 10 s at 60°C, and 10 s at 78°C. In addition to fluorescence readout at 78°C in each cycle, melting curves were obtained.The multimarker RT-qPCR AdnaTest recently –73. The n = 14) using individual cut off values for each gene -Sample Cq(gene)]-[Cutoff(PTPRC)-Sample Cq(PTPRC)]. The transcripts PSA, PSMA, AR-FL, AR-V7 were independent of a growing number of leukocytes, thus, the ΔCq value was calculated as follows: ΔCq = [Cutoff(gene)-Sample Cq(gene)]. Only positive Δ(Δ)Cq values were considered as evaluable signals and signals were analyzed binary to be interpreted as overexpression yes/no results. Results of primer that showed Cq values below 35 in the negative control and results of amplicons with the wrong melt temperature [ΔTm (positive control – sample) >2°C] and [Tm <76.6°C] were excluded. We evaluated a sample to be positive for one transcript, if at least one of the two sample duplicates showed a Cq value below the cut-off. Cq values of all patient samples and healthy donors are listed in CTC isolation was conducted in duplicate from 2 × 5 ml blood for each patient sample. cDNA was analyzed separately from these duplicates. The fluorescence threshold of 0.48 was employed for all transcripts (programmed with StepOne Software v2.3) and defined the PCR cycle used for transcript quantification. CTC expression data was normalized to data of healthy donor controls (n = 2) who had no documented BC associated death. In addition, a univariate and multivariate Cox proportional hazard analysis was conducted to confirm the Kaplan Meier findings and to identify factor dependencies. P < 0.05 were considered to indicate a statistically significant difference. Diagrams were computed with the R script mentioned or with Microsoft Excel .Statistical analysis was performed using R (version 3.6.1) with R packages shiny, hmisc, ggplot2, survival, broom, and dplyr. Survival intervals were screened from the time of first diagnosis until the date of recurrence (PFS) or death (OS) and calculated with Kaplan-Meier estimator (Log-rank test). In this cohort, recurrence was supposed in all deceased patients . The patients/participants provided their written informed consent to participate in this study.SK-B and RK supervised the study. SK-B developed the concept and design of the study. SH established and developed the method. CK established, validated the method in the Department of Gynecology and Obstetrics, University Hospital Essen, and wrote sections of the manuscript. OH and A-KB recruited the patients. SK-B, CK, OH, and A-KB collected the experimental and clinical data. SK-B, CK, and SH evaluated, analyzed the data, reviewed, and edited the manuscript. CK and SH performed the statistical analysis and visualized the results. SK-B and A-KB wrote the first draft of the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version.SK-B is a consultant for QIAGEN and has received honoraria from Novartis. CK received support for travel expenses from QIAGEN. OH received honoraria from Roche, Amgen, Pfizer, MSD, and Novartis. SH is an employee at QIAGEN. RK has received honoraria/is part of the advisory board from/at Tesaro, Astra-Zeneca, Medtronic in the last 3 years, and council of IGCS, president of SERGS and proctored and presented for Intuitive Surgical. A-KB received honoraria from Roche.


Spotted lanternfly (SLF) is an invasive insect in the Northeastern U.S. projected to spread nationally and globally. While SLF is a significant pest of vineyards, little is known about the pest in grape agroecosystems including its spatial ecology. SLF spatial patterns were analyzed using a combination of approaches including generalized linear mixed effect models, Moran’s I statistic for spatial clustering, and Empirical Bayesian Kriging. Analysis revealed that SLF displayed significantly clumped distributions in monitored vineyards. Approximately 54% and 44% of the respective adult and egg mass populations were observed within the first 15 m of the vineyard edge. Importantly, the spatial concentration of adults at the edge was consistent temporally, both between years and weeks. Moreover, high populations of SLF on vines were significantly correlated with reduced fruit production in the following year. Mark-release-recapture of SLF revealed that higher proportions of SLF were recaptured on vines with high pre-existing SLF populations, indicating that SLF may exhibit aggregation behavior along vineyard perimeters. Monitoring and management efforts for SLF should be prioritized around vineyard edges as it may significantly reduce infestations and subsequent damage. Spatial ecology offers unique insights into how organisms organize and utilize habitats across both space and time. Geostatistical and spatial analyses identify predictable invasion patterns of invasive pests, which aid in describing how these species colonize foreign habitats and what landscape attributes contribute to the their success8. Importantly, the use of spatial ecology in pest management can identify vulnerable ecosystems  as well as inherent weaknesses in the pest’s ecology which may be exploited in management efforts.The spatial ecology of invasive species has received considerable attention in the past decadeLycorma delicatula (White) (Hemiptera: Fulgoridae), is a new invasive insect in the Northeastern United States native to China9. Prior to SLF’s introduction to the US, it has become invasive in Korea and Japan11. Owing to its broad host range, high fecundity, and inconspicuous egg masses, SLF is expected to invade regions throughout North America and into new countries13. SLF has been reported to feed on 70 different plant hosts, including many valuable ornamental and agricultural crops9. Damage caused by SLF and corresponding efforts to offset feeding damage are expected to cost millions in the United States. Recent economic analyses from Pennsylvania indicate that SLF infestations have already amounted to an annual economic loss of nearly 50.1 million dollars (USD) statewide14.Spotted lanternfly (SLF), 15. SLF adults and nymphs feed on plant phloem from the trunk, cordon, or shoot tissue15 which likely reduces photosynthetic potential and vigor of vines. Damage caused by these insects may significantly alter grape production. In a recent 2020 survey, most grape growers (53%) stated that they would forgo replanting or expanding their production without effective SLF management options . The need for effective management strategies is paramount to securing the livelihoods and industries impacted by SLF.Grape production is greatly impacted by SLF infestations. In the Northeast, growers report reduced vine production or complete plant loss with poor SLF control. Numbers of SLF on individual vines can easily exceed 400 adults, with seasonal averages of 7.5 SLF per vine16 and agrichemicals17, predation from natural enemies18, and chipping wood that contains egg masses19. Currently, pest management for SLF in grape has relied heavily on repeated insecticide applications. Unsurprisingly, the number of insecticide applications in SLF infested vineyards has increased drastically, with most vineyards increasing applications by 2–5 times as compared to pre-SLF infestation20. SLF is expected to continue to be a serious threat to many agricultural and ornamental industries. However, the success of SLF management tactics relies on appropriate timing and placement. Therefore, it is critical that research addresses not only effective management options but also the biology underpinning the success of these management tactics.Some studies have identified approaches to managing SLF infestations including applications of entomopathogens23, no studies have identified the geostatistical or spatial patterns of SLF in grape production. The objectives of this study were to characterize SLF adult and egg distributions across space and time within commercial vineyards in Pennsylvania. Using a combination of weekly monitoring data and grid sampling from 6 to 9 Pennsylvania vineyards, we examined the spatiotemporal relationships of SLF in grape agroecosystems. Mark-recapture of adult SLF was also used to evaluate dispersal behavior based on existing abundance of SLF within vineyards. Further, vine health metrics were measured to determine associations with SLF infestation. We hypothesized that SLF would aggregate around vineyard edges, with greatest densities of SLF adults and egg masses within the perimeter of vineyards. We also predicted that SLF would be negatively associated with vine health.Management of SLF could significantly benefit from an improved understanding of the spatial patterns and damage caused by this pest. Scouting and monitoring can be targeted to specific problem areas, and insecticide use tailored to areas where infestations are highest. Despite a number of recent studies examining basic biology, management tactics, and monitoring efforts2 = 237.3, df = 5, p < 0.0001). Approximately, 54% of SLF were found within the first 15 m of the vineyard edge , SLF distribution was significantly impacted by the interaction between grape variety and distance from vineyard edge . Greater proportions of SLF were observed within the first two distances on hybrid vines . However, the pattern was similar for both small (> 0.5 ha) and large (> 1.5 ha) vineyard blocks, and SLF proportions decreased from the vineyard edge. . Similar to the findings reported for adults, 44% of SLF egg masses were found within 15 m of the vineyard edge and a significantly lower proportions of egg masses (12%) were found on vines located 75 m from the vineyard block edge . Reduced fruit clusters were associated with high numbers of SLF in the previous growing season (adjusted R2 = 0.44) . Other health metric measures  were not associated with adult SLF infestation in the previous year .Adult SLF numbers were significantly negatively associated with two plant health metrics: cluster production and leaf redness . On average, 2 times more adult SLF were found within the first 15 m of the vineyard edges compared to the interior of the vineyard. Distance from the wood edge also significantly influenced egg mass abundance . On average, 2.8 times more SLF egg masses were found within the 15 m of the vineyard edges compared to the interior of the vineyard block. SLF across all vineyards were significantly spatially autocorrelated using Moran’s I statistic with the Moran’s index ranging from 0.19 to 0.68 . These results were consistent at both 12 and 24 h .Pre-existing SLF populations on vines significantly impacted the percent of SLF recaptured after both 12 and 24 h Fig. . Vines wSLF is a noted pest of vineyards in the Northeastern U.S., however, little is known about its biology, behavior, and management in these agroecosystems. In this study, we evaluated the spatial and temporal relationships of SLF in grape systems. In line with our proposed hypothesis, we found that the greatest proportions of SLF adults and egg masses were consistently observed along the vineyard perimeters. Further, greater densities of SLF were significantly related to reduced vine health (reduced fruit production and increased leaf redness). This is the first study to report on the spatial organization of SLF within a landscape as well as indicate the potential negative effects of SLF on grapevine yield.15. Therefore, these relationships are likely significantly impacted by the quantity and quality of nearby host plants. Indeed, in this study we found that vineyard distance from wooded edges was strongly correlated with both SLF adult and egg abundance in the vineyard. Successful SLF control will likely necessitate some type of landscape management, either at a local or regional scale. Edge effects have been observed for other agricultural insect pests (see review24) with predictive models that attempt to explain such behavior25, which presents an opportunity to exploit this spatial organization for precision pest management. Further research may also address specific landscape factors that explain the prevalence of SLF in certain ecosystems, including land cover, which was not analyzed in this study.Weekly monitoring and grid sampling of SLF in vineyard blocks revealed clumping of SLF around vineyard edges. From geostatistical modeling using the EBK approach, higher densities of adult SLF were predicted at the outside edge of vineyard blocks and low densities in the vineyard interior. Importantly, this occurs regardless of overall population density within the vineyard. These edge effects are likely impacted by landscape features and the ability of SLF to utilize resources in those landscapes. SLF is not confined to vineyards and is known to disperse into vineyards from wooded edges18, thus patterns observed in this study may be a result of large daily influxes to vineyards15. Insecticide exposure could also cause sublethal effects on SLF (e.g. reduced mobility). Therefore, it is possible that SLF may assume a different spatial distribution in unmanaged or infrequently sprayed vineyards. While egg masses displayed a clumped distribution at vineyard edges, this pattern had a higher degree of variance compared to adult distributions. Further study is needed to elucidate the biological and/or ecological mechanisms that explain this discrepancy. Our previous study indicated that SLF densities decrease in vineyards towards the end of the growing season15 which may reduce overall ovipositional activity. Female SLF may prefer to oviposit in nearby wooded edges, thereby reducing egg laying in vineyards and decreasing edge effects associated with egg masses.All vineyards monitored in this study used repeated insecticide applications to manage SLF which could alter their spatial organization. Insecticides can be highly efficacious against SLF26. Other vineyard pests, such as grape berry moth and Japanese beetle also exhibit edge effects and may increase the utility of these targeted management efforts (e.g. perimeter sprays) in grape production27.Our findings suggest that insecticide regimes can be optimized to control SLF populations. Specifically, vineyard edges infested with SLF can be targeted with insecticidal sprays (e.g. perimeter sprays) which will reduce overall insecticide use and cost and mitigate negative off-target effects. Furthermore, management that focuses on the reducing SLF populations at vineyard edges may prevent infestations in the vineyard interior. Non-chemical approaches, like netting along vineyard edges, may limit SLF dispersing into vineyards. These tactics have been successful with highly mobile insects including brown marmorated stink bug28. If SLF feeding reduces overwintering resources for the vine, this could manifest as increased damage to primary buds rather than complete bud death. This may explain why our study failed to find a significant effect of SLF abundance on the prevalence of dead shoots. Red leaves on red producing cultivars were significantly related with SLF abundance, which could be used as a metric to predict stressed plants and inform potential changes in sap flow and nutrient status of the vine29. Crown gall was not significantly related to SLF abundance; however, our study did not include measurements of crown gall in the vineyards prior to SLF infestations, which limits our ability to address this relationship. Further study is needed to evaluate the relationship between SLF feeding injury and the severity and spread of crown gall. Additionally, nymphal SLF were not evaluated in this study and may influence these vine health metrics, although low levels of this life stage have been documented in vineyards15.The number of clusters per shoot were negatively associated with increased adult SLF from the prior year. Primary buds, which bear the majority of fruit, are the most sensitive to cold damage, followed by secondary buds (reduced fruit) and tertiary buds (no fruit)Our mark and recapture results suggest that SLF may aggregate in areas with higher densities of other SLF individuals. Thus, the relationships presented here are likely not only a result of vineyard management but also SLF behavior. Advantages of feeding in groups including overcoming plant defenses or mate selection may explain these patterns. These results should be further explored as they could play a critical role in developing mass-trapping, monitoring, or behavioral disruption tactics. It is important to note that the potential effects of powder-marked SLF on their mobility has not been fully evaluated and this should be studied in greater detail. Furthermore, this study only examined the short-term retention of SLF within an area (12–24 h), and longer time periods (7–14 days) may provide meaningful insights to behavior and management of SLF.The spatial concentration of SLF adults at vineyard edges was consistent temporally, both between years and weeks of sampling. These findings suggest a predictable trend in behavior which can be used to optimize monitoring and precise targeting of management efforts, including release of natural enemies and chemical applications in these agroecosystems. Utilizing this knowledge to modify chemical applications could reduce overall insecticide input into the production system, reducing non-target effects and secondary pest problems.Vitis vinifera varieties in all vineyards, and a subset of vineyards (n = 4) also included monitoring on hybrid Vitis cultivars. Vineyards were actively managed for SLF and other pests with insecticide applications.SLF infestations were sampled weekly in vineyards within southeastern Pennsylvania in 2018 and 2019. Six vineyards were used in both 2018 and 2019, plus an additional 2 vineyards were added in 2019. Weekly monitoring for adult SLF began in September in 2018 and August in 2019. In both years, monitoring continued until no more SLF were observed in each vineyard. At least two rows were randomly selected within each vineyard block and the number of SLF were recorded every 15 m along a transect spanning the entire length of the row. The length of each row varied depending on the vineyard (50 m–210 m). At each distance, the total number of SLF on the vine were counted. SLF was monitored on 29, which may be linked to phloem feeding by SLF. In the fall of 2019, percent leaf redness was estimated on a scale of 0–100% for all monitored vines. Leaf redness was only observed on red fruiting cultivars . Similarly, crown gall severity at the trunks and cordon were taken for each vine due to the association of crown gall (caused by the bacterium Agrobacterium vitis) and grapevine decline, which can be exacerbated by physical injury to the plant30. Severity was ranked on a scale of 0  to 5 . For analysis, the severity rankings for both cordon and trunk were combined.In grapevines, the fruit set in a given year is determined in the year prior, when buds are being formed. After overwintering, these buds provide shoots and clusters for the following season. In spring 2019 and 2020, the number of dead shoots, living shoots, and clusters on each shoot were counted on all monitored vines to evaluate the relationship between SLF infestation and bud fruitfulness and survival. Leaf redness in grapevines has been related to reduced translocation and accumulation of sugars within the canopyAdult and egg mass counts were completed in selected blocks within 7 different vineyards in southeastern Pennsylvania. Vineyard block size varied with the smallest being 0.4 hectare and the largest 1.5 hectares. Blocks were often mixed cultivar plantings and varied in SLF infestation pressure. Adults were sampled between 16 and 27 Sept 2019 and egg masses were sampled from 7 to 16 Oct 2019. Within each block, vines were selected every 5 m along each row and the total number of SLF adults and egg masses counted.Adult SLF (n = 348) were collected from unmanaged forested areas and marked with fluorescent powder . SLF were randomly split into groups of 20 and released within vineyard rows featuring variable pre-existing densities of SLF. Prior to the release of the marked SLF adults, densities of SLF in experimental vineyard rows were determined by counting a subset of vines in each row. Marked SLF were placed at the base of vines in each vineyard row. After either 12 or 24 h, vines were examined, and the number of remaining of marked SLF counted. The powder was tested prior to the initiation of the experiment on adult SLF to ensure it would not lead to increased mortality after 48 h of exposure.31). The proportion of SLF at each distance was determined from the total number of SLF counted in that row. Proportion of SLF at each distance was analyzed using fixed effects of distance and site, and year, week, and row as random nested effects using a binomial distribution. In order to determine the effect of vineyard edges on the proportion of SLF, a ‘distance’ variable was assigned to each sampling point within a vineyard transect to indicate the distance from the edge of the vineyard block. Additional analyses tested the effect of vineyard block size and grape variety as fixed effects. Due to the potential variation of grape variety across the different vineyards, only vineyard blocks that contained both V. vinifera and hybrid varieties (4/8 vineyards) were included in the analysis with grape variety.Weekly monitoring data were analyzed using generalized linear mixed models (GLMM) (R version 3.6.2)  or a normal distribution . Health metrics were modeled with previous SLF infestation (average adult SLF per vine) as the fixed effect whereas vineyard and year were considered random effects in model. Mark and recapture data were analyzed assuming a binomial model where the total number of recaptured SLF was compared to the total number of SLF released. Adjusted R squared values were computed for all significant models using R package, MuMIn35. A Moran’s I Index value of 1 signifies strong spatial autocorrelation (clustering), 0 indicates no spatial autocorrelation (random), and -1 indicates highly dispersed autocorrelation. Empirical Bayesian Kriging (EBK) was also used to observe spatial trends in abundance of egg masses and adult SLF within each vineyard block. EBK is a mapping method to predict unmeasured values using measured values within a spatial framework36. For all EBK models, the power function was used to create the semiovariograms.Grid sampling data was georeferenced with latitude and longitude using Google Earth. Using ArcGIS Pro, a polyline was drawn around the wood edge for each vineyard, and the distance of each grid sampling point was then calculated to the nearest wood edge . The distance from the wooded edge and the number of SLF (adults and egg masses) were then analyzed using a GLMM with row and vineyard as random nested effects using a Poisson distribution in R. The number and position of SLF in each vineyard were also analyzed in ArcGIS Pro using Anselin Local Moran’s I tool to determine spatial relatedness of pointsSupplementary Information.


Worldwide mortality due to stroke and heart disease will increase to 23.6 million in 2030 . BulgariThe scope of this study was to assess the role of rehabilitation for improving the functional capacity, independence in the ergotherapeutic activities and the quality of life of the patients with stroke, as well as the necessity of a lasting and adequate to the patients’ needs rehabilitation. The patients had two seven-day hospital treatment courses in the Physical Medicine and Rehabilitation Clinics at the General Hospital “St. Panteleymon” Plovdiv, Republic of Bulgaria for the period between 2016 and 2017.The prescription period for the studied patients was from one to ten months. Forty-three patients had two full treatment courses as between the treatment stages 4 patients dropped out because of the neurological complications that occurred. It was of great interest to us to trace the functional recovery of patients with relatively the same localization of the brain damage, which was in correlation with the rehabilitation potential and the complex rehabilitation programme.In the acute stage of the disease, the physical therapy and rehabilitation are targeted at preventing complications in the locomotor system , of the nervous and respiratory system . As a reThe choice of specific kinesitherapeutical techniques , was done individually for each patient after precise kinesic analysis of the spasticity syndrome and functional assessment in the beginning and the end of each rehabilitation programme. An individual complex rehabilitation algorithm was used, which includes kinesitherapy and therapeutical massage: inhabiting positions under the Bobath approach for suppression of the spastically increased muscle tonus; exercises to decrease the muscle imbalance; stretching exercises; suspension of upper and lower limbs; methods for proprioceptive neuromuscular facilitation in accordance with the Kabat, Knott and Voss approach for neuromuscular stimulation; diagonal-spiral patterns for upper and lower libs of Kabat; exercises for coordination and balance .The applied complex functional rehabilitation, combined with personal health care led to significantly faster reduction of the functional dysfunctions and improvement of the independence and the quality of life of the patients. However, the recovery is not sufficient in terms of the independence of the general movements and social re-integration. The patients and their relatives must be trained to apply continuously the rehabilitation activities along with the ergotherapeutical at home. The tolerant attitude on behalf of close people and relatives is essential for people with stroke after the incident. The significance of training in the complex approach is also very important for the improvement of recovery skills and the functional condition which leads to improvement of quality of life in patients with stroke.


The expression of the androgen receptor (AR) and its splice variant AR-V7 is crucial for prostate cancer (PCa) biology. An immunohistochemical staining was performed on a tissue microarray with specimens from 410 PCa patients. AR staining, neither in the nucleus nor in the cytoplasm was associated with prognosis. AR-V7 staining of the general cytoplasm was associated with a shorter relapse free survival (RFS), whereas AR-V7 staining of cytoplasmic granules was associated with a longer RFS. Further subgroup stratification for AR-V7 granular staining revealed it as an independent prognostic factor in younger patients (age ≤ 65), patients with negative CK20 staining and patients with perineural invasion. Altogether, AR-V7 protein detected in granular cytoplasmic structures is an independent prognostic factor for RFS in PCa patients.p = 0.006). In a further subgroup analysis by multivariate Cox’s regression analysis, AR-V7 was an independent prognostic factor in the following groups: age ≤ 65 , negative CK20 staining , and positive perineural invasion . Altogether, AR-V7 protein in granular cytoplasmic structures is an independent prognostic factor for RFS in PCa patients.Prostate cancer (PCa) is the second most common cancer, causing morbidity and mortality among men world-wide. The expression of the androgen receptor (AR) and its splice variants is a crucial factor of prostate cancer biology that has not been comprehensively studied in PCa tumors. The aim of this study was to characterize the protein expression of the AR and its splice variant, AR-V7, and their subcellular distributions in PCa by immunohistochemistry and to correlate the results to the clinicopathological data and prognosis. Immunohistochemical staining for AR and AR-V7 was performed on a tissue microarray (TMA) with specimens from 410 PCa patients using an immunoreactive score (IRS) or only the percentage of AR-V7 staining in cytoplasmic granules. Nuclear or cytoplasmic AR staining was not associated with prognosis. AR-V7 staining was only occasionally observed in the nucleus. However, AR-V7 staining in the cytoplasm or in cytoplasmic granules was associated with relapse-free survival (RFS). AR-V7 staining of the cytoplasm was associated with a shorter RFS, whereas AR-V7 staining of cytoplasmic granules was associated with a longer RFS. In a multivariate Cox’s regression analysis, only negative (<5%) AR-V7 staining of cytoplasmic granules remained an independent prognostic factor for RFS (HR = 5.3;  Prostate cancer (PCa) represents the second most common cancer and the fifth most common cause of cancer-associated death in men globally . PCa is Based on the dependency of PCa on AR signaling, most therapeutic approaches affect the androgen receptor itself, androgen biosynthesis and/or the interaction of the AR with androgens. As a consequence, the tumor develops therapeutic resistance to AR-targeted therapies due to AR mutations, AR amplification or AR splice variants ,10,11. AThe aim of the present study was to examine AR and AR-V7 protein expression and their subcellular distributions in PCa and to correlate the findings with clinicopathological and prognostic data.A TMA containing prostate tumor sections from a cohort of 410 PCa patients was evaluated for AR and AR-V7 protein expression by immunohistochemistry (IHC) using an AR-V7 monoclonal antibody as described in the methods section. AR staining was detected in the nucleus and cytoplasm, whereas AR-V7 expression was observed in the cytoplasm (cytoplasmic) or in distinct granular structures in the cytoplasm (granular), but only occasionally in the nucleus, by IHC. The staining distributions for AR and AR-V7 were as follows: For AR cytoplasmic staining, 282 cases (68.8%) were negative, and 127 cases (31.0%) were positive. For AR nuclear staining, 96 cases (23.4%) were negative, and 314 cases (76.6%) were positive. For AR-V7 cytoplasmic staining, 59 cases (14.4%) were negative, and 347 cases (84.6%) were positive. For AR-V7 granular staining, 261 cases (63.7%) were negative, and 144 cases (35.1%) were positive . Howevers = 0.142; p = 0.005), prostatectomy Gleason sum (GS) , relapse occurrence (RFS) , pathological tumor stage (pT) , CK20 staining , AR nuclear staining , and AR-V7 cytoplasmic staining , but it was not inversely or negatively correlated with any clinicopathological or molecular factor (AR cytoplasmic staining was directly (positively) correlated with perineural invasion (Pn) , pT , CK20 staining , AR cytoplasmic staining , and AR-V7 cytoplasmic staining . However, it was negatively correlated with AR-V7 granular staining , prostatectomy GS , pT , CK20 staining , AR nuclear staining , and AR cytoplasmic staining . It was negatively correlated with AR-V7 granular staining , prostatectomy GS , RFS , pT , metastasis occurrence , AR nuclear staining , and AR-V7 cytoplasmic staining  encoded by the cryptic exon 3 (CE3) of AR-V7, that competes with the AR-V7 antibody was applied as previously described . ApplicaThe association of AR and AR-V7 staining in the 410 PCa tumor samples with patient survival was examined by Kaplan-Meier analysis. AR staining in the nucleus or in the cytoplasm was not associated with OS, DSS, nor RFS. In addition, both AR-V7 staining in the cytoplasm and in the cytoplasmic granules were not associated with OS or DSS.p = 0.027; p = 0.057).However, both AR-V7 staining patterns were significantly associated with RFS but had opposite correlations . In Kaplp = 0.263), DSS (p = 0.473), nor RFS (p = 0.164) (data not shown). However, when we considered only the best (both negative) and the worst prognosis groups (both positive), there was a significant difference in RFS  or DSS (p = 0.309). Univariate Cox’s regression analysis revealed that the combination of cytoplasmic AR and cytoplasmic AR-V7 positivity had the same trend as the single cytoplasmic AR-V7 positivity towards being associated with a 6.8-fold risk of relapse occurrence .Since AR cytoplasmic and AR-V7 cytoplasmic staining are related, we were interested if their combination could be associated with prognosis. Combination of both staining patterns resulting in four groups, showed no significant differences in OS .Multivariate Cox’s regression analysis, adjusted for pT, prostatectomy GS and Pn, revealed no association between cytoplasmic AR-V7 positivity and RFS  showed that negative granular AR-V7 staining was an independent prognostic factor for RFS , GS 6 (p = 0.040), age at diagnosis ≤ 65 years (p = 0.001), age at diagnosis > 65 years (p = 0.045), without Pn (p = 0.005), with Pn (p = 0.018), and negative CK20 staining  in the Kaplan-Meier analysis (p = 0.029), negative CK20 staining , and in the group with Pn   .Next, we examined which granular structures were stained for AR-V7. Since the structures resembled the Golgi apparatus, we stained the PCa TMA for GOLGB1 , a major protein of the Golgi apparatus. Interestingly, we observed similar granular structures stained by both AR-V7 and giantin . AlthougThe AR is the most important hormone receptor in PCa, and its splice variants, mainly AR-V7, can participate in endocrine resistance . Here, tBy correlating AR cytoplasmic, AR nuclear, and AR-V7 cytoplasmic staining with clinicopathological data and with each other, we found a positive correlation with perineural invasion (Pn), pathological tumor stage (pT), and CK20 staining and between these three staining patterns . Additionally, AR cytoplasmic and AR-V7 cytoplasmic staining was negatively correlated with the Gleason sum (GS) at prostatectomy, whereas AR-V7 cytoplasmic staining was associated with relapse occurrence. AR-V7 granular staining showed the opposite result, i.e., a negative correlation with the clinicopathological parameters Pn, prostatectomy GS, relapse occurrence, pT, metastasis occurrence, and with AR nuclear and AR-V7 cytoplasmic staining.In our survival analysis, AR staining in the nucleus or in the cytoplasm was not associated with OS, DSS, nor RFS. In addition, AR-V7 staining in both the cytoplasm and the cytoplasmic granules was not associated with OS or DSS. However, positive AR-V7 staining in the cytoplasm was a negative prognostic marker for RFS in univariate analysis, in agreement with the multivariate analysis result by Guo et al. . ConversBased on the assumption that only nuclear AR-V7 can affect gene transcription, several studies scored only nuclear AR-V7 expression and disregarded cytoplasmic staining in their experiments . IncreasWe showed, for the first time, that AR-V7 granular staining (≥5%) is an independent prognostic biomarker for longer RFS in PCa patients. Next, we were interested in whether this prognostic effect could be further stratified into different clinicopathological subgroups. We found that AR-V7 granular staining (≥ 5%) was an independent prognostic marker for RFS in the subgroups age ≤ 65 years, patients with Pn and patients with negative CK20 staining (IRS ≤ 2) but was not in the groups age > 65 years and patients with positive CK20 staining (IRS > 2). Recently, we showed that another marker, i.e., progranulin (GP88) protein expression, was associated with a shorter RFS in younger PCa patients. Since there is an increase in early-onset PCa in Europe and America . Howeverhttps://www.proteinatlas.org/ENSG00000173230-GOLGB1/pathology). We think AR-V7 detected as granular staining means that AR-V7 is not active. We hypothesize that granular AR-V7 is involved in a protein degradation process in the Golgi apparatus. In this way, AR-V7 cannot function as transcription factor and this may slow down tumor growth and tumor development resulting in a longer RFS.Next, we were interested in why AR-V7 granular staining could be associated with a longer RFS. AR-V7 participates in endocrine resistance . TherefoWe showed in this study that the granular staining patterns of AR-V7 and giantin are very similar, suggesting that granular AR-V7 is located in the Golgi apparatus. However, what function could AR-V7 have in the Golgi apparatus? There are several mechanisms for cellular proteostasis, including protein-degradation mechanisms (reviewed in Reference ). RecentThe tissue microarrays (TMAs) contained consecutively collected, formalin-fixed and paraffin embedded tumor samples of 410 PCa patients. They were diagnosed in the Department of Pathology, University Hospital Erlangen between 1999 and 2010. The tumors were collected from radical prostatectomy specimens, and the follow-up time from date of diagnosis ranged from 0 to 246 months (median 95 months). For OS, the observation time ends with the death of the patient , all other patients that were alive at their last visit in the observation time are considered as alive (no event). For DSS, the observation ends either with the tumor-specific death (=event); any other reason for death or patients that are still alive in the follow up time are considered as alive (no event). For RFS, the observation time ends either with a recurrence (=event), all patients who did not develop a recurrence within the observation time are considered as recurrence-free (=no event). The tumor histology was reviewed by experienced uropathologists (AH and ME). All procedures were performed in accordance with the ethical standards established in the 1964 Declaration of Helsinki and its later amendments. All patients starting in 2008 provided written informed consent. For samples collected before 2008, the Ethics Committee in Erlangen waived the need for informed individual consent. The study was approved by the Ethics Committee of the University Hospital Erlangen (No. 3755). Since archival tissue was studied several years after resection, the analysis of AR and AR-V7 protein expression was retrospective without affecting clinical decisions. Clinicopathological data of the patients are shown in 2O2. Visualization of bound antibody was performed using the ultraVIEW TM DAB system (Ventana). All sections were counterstained with hematoxylin II/Mayer’s hematoxylin (Ventana).For analysis of AR and CK20 protein expression, IHC staining was performed on a fully automated Ventana Benchmark Ultra autostainer . The following antibodies were applied: AR expression was analyzed with a monoclonal mouse AR antibody  and CK20 expression was studied with a monoclonal mouse CK20 antibody  by a routine staining procedure as previously described . BrieflyFor the AR-V7 protein expression study, a manual immunohistochemistry (IHC) protocol was applied as previously described . A primaThe expression of AR (nucleus/cytoplasm), AR-V7 (cytoplasm) and CK20 (cytoplasm) was detected by assessing the percentage of stained tumor cells and the staining intensity semiquantitatively. The percentage of positive cells was scored as follows: 1, 1–9% positive cells; 2, 10–50%; 3, 51–80%; and 4, >80% positive cells. Staining intensity was scored as 0, negative; 1, weak; 2, moderate; and 3, strong. The immunoreactive score (IRS) was calculated as the product of staining percentage and staining intensity, resulting in an immunoreactive score (IRS) from 0 to 12 . Negativ2-test. The associations of AR and AR-V7 expression with overall survival (OS), disease-specific survival (DSS), and relapse-free survival (RFS) were determined by univariate  and multivariate  analyses. A p-value < 0.05 was considered statistically significant. The statistical analyses were performed with the SPSS 21.0 software package .The correlations between the IHC scores and clinicopathological data were calculated using Spearman’s bivariate correlation or the chiAR-V7 staining in the cytoplasm or in cytoplasmic granules was associated with relapse-free survival (RFS) in the Kaplan-Meier analysis. However, AR-V7 cytoplasmic positive staining was associated with a shorter RFS, and AR-V7 granular staining was associated with a longer RFS. In a multivariate Cox’s regression analysis, negative (< 5%) AR-V7 granular staining remained an independent prognostic factor for RFS. Further subgroup analysis revealed that AR-V7 granular staining was an independent prognostic factor in the following groups: younger patients (age ≤ 65), patients with negative CK20 staining and patients with perineural invasion. Altogether, AR-V7 protein in granular cytoplasmic structures appears to be an independent prognostic factor for RFS in PCa patients.


Spissistilus festinus (Say) (Hemiptera: Membracidae) was shown to transmit Grapevine red blotch virus (GRBV) in a greenhouse study. Grapevines infected with GRBV exhibit reduced sugar accumulation, altered secondary metabolite production and delayed berry maturation that negatively impacts wine quality and economics. Augmentative biocontrol may be a useful integrated pest management (IPM) tool for suppressing S. festinus populations in vineyards, but minimal research has been conducted on testing potential predators against the different life stages of S. festinus. The susceptibility of S. festinus adults and nymphs (1st through 5th instar) to predation by six commercially available biocontrol agents in petri dish and bell bean plant arenas was determined under greenhouse conditions. No significant mortality of S. festinus nymphs or adults occurred when exposed to Cryptolaemus montrouzieri adults, C. montrouzieri larvae and Sympherobius barberi adults in petri dish or bell bean plant arenas. Significant mortality of 1st and 2nd instar nymphs of S. festinus in the presence of Zelus renardii nymphs was observed in petri dish but not in bell bean arenas. Hippodamia convergens adults and Chrysoperla rufilabris larvae both consumed a significant number of S. festinus nymphs in petri dish and bell bean arenas. No significant predation of S. festinus adults was documented in this experiment. Results of this study aid in identifying predators that may be suitable candidates for additional field testing to determine their potential efficacy as biocontrol agents of S. festinus in a vineyard setting. Grapevine red blotch virus (GRBV) is the causative agent of grapevine red blotch disease (GRBD) –3. GrapeSpissistilus festinus (Say) (Hemiptera: Membracidae) was shown to transmit GRBV to grapevines in a greenhouse study [S. festinus was considered an incidental pest of grapevines [S. festinus has typically focused on other cropping systems such as alfalfa, peanuts, and soybeans [S. festinus in grapevines, recent studies have focused on the biology and behavior of S. festinus in vineyards, on identifying feeding and reproductive hosts and documenting the treehopper’s seasonal dynamics in vineyards [The threecornered alfalfa hopper, se study . Prior tapevines  due to tapevines . In factsoybeans ,15, withineyards –18.S. festinus adults move into vineyards from surrounding habitats in early to mid-February. On arrival to a vineyard, females may oviposit on ground vegetation provided suitable hosts are available [S. festinus [S. festinus inhabit ground cover present in vineyards, insecticides are not a viable control option as candidate insecticides are not registered for application to vineyard ground cover. Instead, resident vegetation and/or cover crops could be tilled under during spring when early instars of S. festinus are present. While springtime tillage is a common vineyard practice [In northern California, overwintering vailable . Legumesfestinus ,15,17,19festinus ,21. As spractice ,23, not S. festinus will move into the grape canopy starting in June, causing girdling damage to grape petioles and shoots and potentially transmitting GRBV. While the number of S. festinus adults moving into the grape canopy increases as the season progresses, insecticide treatments applied to the grape canopy are expected to have limited efficacy as many S. festinus remain on ground vegetation [S. festinus collected from vineyards do not test positive for GRBV until June, suggesting that management actions designed to suppress S. festinus populations should be implemented prior to June.Some adult getation . In addigetation  showed tS. festinus populations in vineyards. Previous tests by Medal et al. [S. festinus nymphs and adults by Geocoris punctipes, Nabis roseipennis, and Orius insidiosus. However, these species only represent a few of the many predacious insects available for purchase from commercial insectaries. In this study, five additional predator species that are commercially available for purchase but have not been previously tested were evaluated. A greenhouse study was designed to determine the life stages of S. festinus most susceptible to predation by the following biological control agents: Hippodamia convergens (Guérin-Méneville) (Coleoptera: Coccinellidae) adults, Cryptolaemus montrouzieri (Mulsant) (Coleoptera: Coccinellidae) adults and 2nd instar larvae, Chrysoperla rufilabris (Burmeister) (Neuroptera: Chrysopidae) 2nd instar larvae, Sympherobius barberi (Banks) (Neuroptera: Hemerobiidae) adults, and Zelus renardii (Kolenati) (Hemiptera: Reduviidae) 1st instar nymphs. Results from this study aid in identifying biological control agents that may be suitable for additional field testing, with the ultimate goal of assisting growers and vineyard managers in choosing appropriate biological control agents to suppress S. festinus populations which may aid in reducing vector-mediated spread of GRBV.Release of biocontrol agents in vineyards is an additional IPM tactic that could be used to reduce l et al.  evaluateS. festinus colony was established in March 2019 from adults collected by sweep net from alfalfa, Medicago sativa, fields in Parlier, CA  and Clovis, CA . The authors have a permit issued from the California Department of Food and Agricultural for collecting S. festinus (permit #3479). Spissistilus festinus were collected from privately owned land with permission obtained from the owners prior to collecting. Field studies did not involve endangered or protected species. The colony was maintained in the greenhouse in insect rearing cages  and reared on a combination of potted bell bean (Vicia faba) and purple vetch . Field collected adults were occasionally added to the colony to increase genetic diversity. Adult and 1st-5th instar nymphs of S. festinus were collected from the colony prior to setting up the assays. Eggs were not tested.An Hippodamia convergens (Guérin-Méneville) (Coleoptera: Coccinellidae), Cryptolaemus montrouzieri (Mulsant) (Coleoptera: Coccinellidae), Chrysoperla rufilabris (Burmeister) (Neuroptera: Chrysopidae), Sympherobius barberi (Banks) (Neuroptera: Hemerobiidae), and Zelus renardii (Kolenati) (Hemiptera: Reduviidae). Hippodamia convergens were collected by sweep net from alfalfa, Medicago sativa, fields in Clovis, CA . All remaining predators were purchased from California based commercial biological control providers. The predator’s life stage chosen to be tested against the different life stages of S. festinus was dependent upon the life stage of the predator available for purchase from commercial insectaries. This decision was made to reflect the predator/prey interactions expected from the life stage supply available to industry purchasers. Hippodamia convergens and S. barberi were tested as adults, C. montrouzieri were tested separately as 2nd instar larvae and adults, C. rufilabris were tested as 2nd instar larvae, and Z. renardii were tested as 1st instar nymphs. Upon delivery, all predators were held individually in 1-ounce condiment cups and starved for 24 hours prior to being introduced into their testing arena with the exception of Z. renardii, which were purchased as eggs and reared on Aphis gossypii for 4–7 days before being subjected to the same 24-hour starvation period prior to testing.Five species of biocontrol agents were tested: S. festinus food source in petri dish arenas, green beans (Phaseolus vulgaris) were cut into ~2.5 cm sections and hot glued to the bottom of the Petri dish (100 mm x 15 mm) to hold the green bean section in place. A crumpled moistened paper towel (17.5 cm2) was placed in each Petri dish as a water source. One S. festinus of a certain life stage (1st through 5th instar or adult) was placed on the green bean in each petri dish and allowed 30 minutes to settle before a predator was introduced into the arena. Each Petri dish was sealed with Parafilm immediately following introduction of the predator. Petri dishes were placed in the greenhouse and S. festinus mortality recorded after 24 hours. Ten replicates of each of the six developmental stages (1st– 5th instar and adult) of S. festinus were tested. To quantify S. festinus background mortality, five Petri dishes per S. festinus life stage that contained food and a water source, but no predator served as controls.To provide an S. festinus held on plants. Bell beans (Vicia faba) were chosen as the test plant because they are a feeding host of S. festinus  and have an upright growth pattern. Bell bean seeds were purchased from Harmony Farms Supply & Nursery, Sebastopol, CA, USA. Professional growing mix potting soil was purchased from Sun Gro Horticulture Canada Ltd., Seba Beach, Canada. Dart red plastic party cups 532-ml  served as pots with holes (~4 mm) drilled in the bottom for drainage. Each pot was planted with one bell bean seed and allowed to grow for 12–14 days to ~10–15 cm in height before being used for testing.As petri dish arenas are a simplified environment, additional tests were conducted with S. festinus of a certain life stage was placed on the base of the bell bean plant and allowed 30 minutes to settle before a predator was introduced into the cage. Each cage was sealed and attached to the pot with 5.1 cm packaging tape  immediately following introduction of the predator. Bell bean cages were held in the greenhouse and mortality recorded after 24 hours. Ten replicates each of six developmental life stages (1st– 5th instar and adult) of S. festinus were tested. Ten bell bean cages per life stage containing prey but no predator served as controls that quantified background mortality. Petri dish and bell bean arenas were conducted simultaneously for each predator tested.Mesh cages were constructed by cutting 3.8 liter plain top paint strainers  in half vertically and sealing the length of the vertical opening with hot glue. Bamboo skewers– 46 cm  were cut to 36.8 cm and three skewers were placed vertically into the pots equidistant from each other in a triangular arrangement. Mesh cages were placed over the skewers. One S. festinus life stage, test arena type, and all possible interaction terms on S. festinus mortality. As predators were not present in controls, the predator species term in the analysis of data collected from controls represented the effect of test date on S. festinus mortality. Replicates in which the predator died before the end of the assay were excluded from analysis .A generalized linear model with binomial error and a logit link function was fit to control and predator data sets using JMP stats . Each anS. festinus life stage combinations that resulted in significant S. festinus mortality, adjusted mortality was determined for each predator species by S. festinus life stage combination using Abott’s formula [S. festinus mortality in the presence of the predator was considered significant. Adjusted mortality values of 100% or 0% were not associated with an estimate of error as there was no variation in the response .To determine the predator species by  formula . SubsequS. festinus held in control petri dishes  died and 30 out of 360 (8.3%) S. festinus held on control bell bean plants died. In controls, S. festinus mortality was not affected by predator species, S. festinus life stage, test arena (petri dish versus bell bean) or any of the interaction terms  on terms . As S. feraction . Thus, SC. montrouzieri larvae  of S. festinus. In contrast, Z. renardii nymphs, H. convergens adults, and C. rufilabris larvae attacked some life stages of S. festinus. None of the tested predators consumed S. festinus adults.The petri dish and bell bean arenas were no-choice assays in which each biocontrol agent was confined to their assigned environment and could either attack/consume the prey provided or not. Predators were unable to choose a different environment or alternate feeding option. The petri dish arena was the most artificial constituting a small area for the predator to search without refuge for the prey. In contrast, the bell bean arena more closely resembled a natural environment with a host plant, soil, and areas for refuge. As petri dish arenas were simplified environments compared to bell bean arenas, petri dish arenas may overestimate the potential of the predator to serve as a biocontrol agent. While bell bean arenas provided a more complex environment, bell bean arenas were still limited in size, confined the prey in proximity to the predator, and lacked alternative prey/predators that may be found in an actual vineyard setting. Nonetheless, both tests aid in determining which (if any) Z. renardii nymphs significantly increased mortality of 1st and 2nd instar S. festinus  and E. elegantula (Osborn) [C. rufilabris may aid in suppressing vineyard pests other than S. festinus.Larvae of n arenas . These r(Osborn) . Thus, rS. festinus populations is promising, especially in no-till vineyards where S. festinus is present. Results from this study suggest that C. rufilabris may be an ideal candidate for augmentative releases. Future research involving choice tests and measuring predation rates as well as field testing of C. rufilabris releases in a vineyard setting and the subsequent impact on S. festinus populations along with the spread of GRBD are needed.The use of augmentative biocontrol to help reduce an insect pest population is an important tool that can be utilized as part of an IPM program. Identifying commercially available predators that may assist in reducing S1 Table(DOCX)Click here for additional data file.


Bone marrow-derived mesenchymal stem cells (BMSCs) represent an alternative to chondrocytes to support cartilage regeneration in osteoarthritis (OA). The sympathetic neurotransmitter norepinephrine (NE) has been shown to inhibit their chondrogenic potential; however, their proliferation capacity under NE influence has not been studied yet. Therefore, we used BMSCs obtained from trauma and OA donors and compared the expression of adrenergic receptors (AR). Then, BMSCs from both donor groups were treated with NE, as well as with combinations of NE and α1-, α2- or β1/2-AR antagonists . Activation of AR-coupled signaling was investigated by analyzing ERK1/2 and protein kinase A (PKA) phosphorylation. A similar but not identical subset of ARs was expressed in trauma  and OA BMSCs . NE in high concentrations inhibited the proliferation of both trauma and OA BMCSs significantly. NE in low concentrations did not influence proliferation. ERK1/2 as well as PKA were activated after NE treatment in both BMSC types. These effects were abolished only by propranolol. Our results demonstrate that NE inhibits the proliferation and accordingly lowers the regenerative capacity of human BMSCs likely via β2-AR-mediated ERK1/2 and PKA phosphorylation. Therefore, targeting β2-AR-signaling might provide novel OA therapeutic options. Thus, in sympathetically innervated tissues, such as the bone marrow or the synovium and the synovial fluid, NE can be present in a concentration range from 10−9 to 10−5 M [−7 M) NE binds preferentially to α-ARs leading to decreasing cAMP concentrations and subsequently to inhibition of protein kinase A (PKA) [−6 M), β-ARs are primarily activated resulting in cAMP increase and PKA activation . In addition, α- and β-ARs activation alternatively leads to extracellular signal-regulated kinases 1/2 (ERK1/2) pathway activation [Mesenchymal stem cells (MSCs) are multipotent progenitor cells with high proliferative activity ,2. Their−9 M) . In addiage area . Furthero 10−5 M . It was o 10−5 M ,15,16. No 10−5 M : At low  A (PKA) . At hightivation ,19. Prevtivation . Similartivation . Furthertivation , the chotivation . Neverth2) was investigated by RT-PCR (mRNA levels). To recognize autocrine effects, gene expression of TH was also analyzed. Trauma and OA BMSCs expressed distinct subtypes of ARs on day 0 of monolayer culture. In trauma BMSCs, α2B-, α2C- and β2-AR were strongly expressed  an increase of cell numbers was clearly visible until day 7 in both trauma and OA BMSCs cultures . The cel−9 to 10−5 M) for 7 days in monolayer culture under physioxic conditions. After 7 days, BMSCs treated with high concentrations of NE (10−5 to 10−6 M) showed an apparent microscopically visible decrease of confluency. Treatment with NE led to a clear and dose-dependent reduction in cell numbers on day 7 compared to the untreated control group (−5 to 10−6 M) significantly reduced cell numbers in both trauma (−9 to 10−7 M) had no effect on cell numbers. Cell viability of trauma and OA BMSCs, measured by lactate dehydrogrenase (LDH) activity, was not affected by any treatment.BMSCs derived from trauma and OA patients were treated with NE in different concentrations  in comparison to the untreated control group, while the expression of α1A-, α1B- and β2-AR did not change significantly. In contrast, treatment with NE in low concentrations (10−8 M) had no influence on the AR expression profile of OA BMSCs.We investigated if NE treatment over 7 days led to alterations in AR gene expression of trauma and OA BMSCs . In trau−6 M) for 5, 10, 30 and 60 min. NE treatment resulted in an increase of ERK1/2 and PKA phosphorylation already after 5 min  in combination with specific AR antagonists for 7 days in monolayer culture under physioxic conditions. The α1-AR-antagonist doxazosin , the α2-AR-antagonist yohimbine  as well as the β2-AR-antagonist propranolol  were applied. Treatment of both trauma and OA BMSCs with NE in combination with propranlol led to a significant increased cell count compared to NE treatment alone (−8 M) and the specific β2-AR-agonist formoterol  were applied for 7 days. In both trauma and OA BMSCs, treatment with formeterol resulted in a significant reduction of the cell number to the same extent as NE in high concentrations (10−5 to 10−6 M) when compared to the untreated control group  decreased the cell count significantly in comparison to the untreated control group. Cytotoxicity expressed as LDH activity was not affected by any treatment indicating that decreased cell counts are due to reduced proliferation potential and not the result of cytotoxic effects. In contrast, Han et al. as well as Kido et al. demonstrated that the treatment of rat BMSCs with NE in high concentrations (10−4 to 10−7 M) lead to an increased proliferation [First, we determined the AR gene expression profile of human trauma and OA BMSCs because the expression of diverse subtypes of AR is a prerequisite for any effect of NE on these cells. We observed that both trauma and OA BMSCs, do express α2A-, α2B-, α2C-, and β2-AR, while α1A-, α1B-, α1D- β1- and β3-AR were not detected on day 0. This is in line with an earlier study in which α1- and β2-AR were identified immunohistochemically in BMSCs obtained from trauma patients . In contferation ,33. One feration ,35. It mferation . Furtherferation . Also diferation . Similarferation ,40,41. Tferation .−6 M) resulted in a slight but not significant α2A- and α2B-AR down-regulation in OA BMSCs, but especially β2-AR expression did not change after treatment with NE. In contrast, AR expression profile of trauma BMSCs was not affected by treatment with NE. On the one hand, these data indicate that the pathophysiological microenvironment of OA damaged joint tissues might result in the fact that MSCs are more affected by an imbalance of external factors such as growth factors, inflammatory mediators or hormones [To evaluate if the inhibitory effects of NE on the proliferation capacity are a result of AR regulation, we investigated the AR gene expression profile of OA and trauma BMSCs treated with NE. Treatment with NE in high concentrations . Therefore, targeting specific adrenergic receptors might represent a promising approach for the development of future therapeutic options for both trauma-induced or OA-related articular cartilage regeneration.p < 0.001).Human bone marrow was harvested from the iliac crests of patients after knee trauma and from OA patients undergoing total joint replacement. Knee trauma patients  underwent surgery 2–3 weeks after injury upon the initial swelling and hematoma abated. The Ethics Committees of the University of Regensburg (vote number 13-101-0135) and of the Goethe University Frankfurt am Main (vote number 148-17B) approved the project. Experiments were performed in consideration of relevant guidelines and regulations. All patients provided their written consent after being informed about the purpose of the study. A total of 31 patients as BMSC donors were recruited for the study. Patient characteristics are shown in ®® life technologies™, Thermo Fisher Scientific, Darmstadt, Germany), supplemented with 10% fetal bovine serum for mesenchymal stem cells  and 1% penicillin/ streptomycin . Cells were cultured at 37 °C under physioxic conditions  [®® solution , counted manually and seeded for proliferation experiments as described below (see 4.3.). Stem cell-specific surface markers of MSCs were investigated in cells of randomly selected donors by standardized FACS analysis of CD73, CD90, and CD105 (positive markers) as well as HLA-DR, CD11b, CD19, CD34, and CD45 (negative markers) as suggested by The Mesenchymal and Tissue Stem Cell Committee of the International Society for Cellular Therapy and as described by us earlier [BMSCs were isolated from the bone marrow as described previously . MSC sep 5% CO2) , medium  earlier ,55,56. B2, 5% CO2). We used both trauma and OA BMSCS at passages 2–4. Medium was changed after 24 h and supplemented with NE ((±)-Norepinephrine (+)-bitartrate salt, Sigma-Aldrich, Munich, Germany) in different concentrations (10−9 to 10−5 M), the specific β2-AR agonist formoterol  or the specific α2-AR-agonist UK14,304 . To investigate if effects are reversible, cells were treated with NE (10−6 M) in combination with the specific α1-AR-antagonist doxazosin , the specific α2-AR-antagonist yohimbine  or the specific β-AR-antagonist propranolol -(−)-Proporanolol hydrochloride, Tocris Bioscience, Bristol, UK) for seven days in further experiments. Medium was replaced and freshly supplemented every two days. Untreated BMSCs served as control group, where the medium without any agonist or antagonist was replaced every two days and the total medium volume was adjusted to the total volume of the treated groups.BMSCs were seeded at a density of 2.667 cells/cm² and allowed to adhere overnight at 37 °C under physioxic conditions . Cells were cultured for seven days and detached as described above. Total cell count and the number of living cells were determined and compared to an untreated control group. To investigate cell viability, supernatants were obtained on day 7 and used immediately for cell viability assays as described below.To demonstrate possible cytotoxic effects, cell viability was determined by measuring LDH activity in cell culture supernatants using the TaKaRa LDH Cytotoxicity Detection Kit MK401  according to the manufacturer’s instructions. Supernatants were harvested on day 7. In addition, culture medium without cells and dead control treated with 1% Triton-X-100  for 10 min were used as controls. Samples of each donor were analyzed in duplicate in a 96-well plate at 490 nm  2016).−6 M for 5, 10, 30 and 60 min at 37 °C and physioxic conditions  and the phosphorylation of the two major AR-related pathways PKA and ERK1/2 was investigated (single replicate per donor). At least 6 × 104 cells were placed in six well plates and allowed to adhere overnight. After 24 h medium was replaced to remove non-adherent cells. In addition to an untreated group, medium was supplemented with NE 10−6 M. In further experiments, NE 10−6 M was combined with 10−7 M doxazosin, 10−7 M yohimbine or 10−6 M propranolol. Cells were incubated for 10 min at 37 °C under physioxic conditions . Subsequently, wells were washed three times with DPBS . Cells were harvested using PhosphoSafe™ Extraction Reagent according to the manufacturer’s instructions . A 10% SDS-PAGE-gel was loaded with the samples and electrotransferred to a polyvinylidene difluoride membrane . Membranes were blocked in 5% BSA for 1 h at room temperature. Afterwards membranes were incubated with primary antibodies for at least 16 h at 4 °C. Primary and secondary antibodies are listed in ® ChemiDocTM XRS+ with Image LabTM Software . Quantification was made with ImageJ 1.52a  with GAPDH as loading control.To identify AR-dependent signaling pathways cells were treated with NE 10®® RNA/ Protein kit  according to the manufacturer’s instructions. RNA concentration was determined with NanoDrop One spectrophotometer. cDNA synthesis was performed using the qScript™ cDNA Synthesis Kit  in a thermocycler qTOWER3 G  using qPCRsoft 3.4 . Samples were incubated at 22 °C for 5 min, at 42 °C for 30 min, at 85 °C for 5 min, followed by cooling at 4 °C for 60 min.RNA was isolated using NucleoSpinHuman primers were obtained by Thermo Fisher Custom Primers , sequences are listed in Human GAPDH served as housekeeping gene, TH gene expression was investigated to consider possible autocrine effects. PCR was performed in a thermocycler qTOWER3 G , programmed with qPCRsoft 3.4  under following cycling conditions: at 94 °C for 0:01 min, followed by 36 cycles at 94 °C for 0:30 min, 64 °C for 0:30 min and 72 °C for 0:59 min. To analyse gene expression of ARs real-time RT-PCR was performed using TaqPCR Master Mix Kit  on day 0 and day 7 (duplicates per donor). 10 µL TaqPCR Master Mix, 5 µL RNAse free water, 2 µM of each primer and 10 ng cDNA were added to a final volume of 20 µL. ® ChemiDocTM XRS+ with Image LabTM Software . Bands were standardized to GAPDH used as endogenous control.Samples were loaded onto a 1.8% agarose gel subsequently stained with GelRed Nucleic Acid Gel Stain . Bands were detected and quantified with Molecular Imagerp values ≤ 0.05 were considered statistically significant. Experiments were performed with samples of 3–13 patients per experiment using n = 6–7 samples for proliferation experiments under NE influence, n = 4 samples for AR gene expression screenings, n = 4–7 samples for proliferation experiments under NE plus antagonist influence , n = 2–4 samples for intracellular signaling analysis via Western blot . Detailed information on replicates is given in the figure legends.Data were analyzed with SigmaPlot software  via Kruskal-Wallis ANOVA on ranks using the post hoc tests Bonferroni or Mann-Whitney U-test. We used Mann-Whitney U-test to analyze the data between two independent groups.


It is intended as a textbook for students and trainees at various levels of training, including medical physics graduate students, clinical radiation oncology and physics residents, and students in medical dosimetry and radiation therapist educational programs. Teaching and learning medical physics are both challenging tasks. While several textbooks are available for medical physics graduate students, such as the classic Khan's The Physics of Radiation Therapy, there is a limited selection for students and instructors involved in the training programs of other professionals in radiation oncology with limited physics and mathematics backgrounds. This book fills in this gap and is designed to support clinical residents, medical dosimetrist, and radiation therapist trainees who are taking their first medical physics course. The text presents physics principles and knowledge in basic language with interactive video tutorials for a broad audience. For medical physics trainees, it is not designed to replace existing textbooks of a graduate‐level medical physics course, but it serves well as a great study review resource for graduate students as well as for board exam preparation.As suggested by its title, 2https://routledgetextbooks.com/textbooks/9781138591707/interactive‐videos.php). Each video is 10‐15 minutes in length and corresponds to a specific topic in the text. These videos mimic an interactive classroom learning environment with whiteboard drawings and embedded quiz questions that can be invaluable to enhance the learning experience and effectiveness.This book has over 350 pages and consists of 27 chapters. These chapters are organized and structured in a logical sequence beginning with basic radiation physics principles and progressing to various physics aspects involved in modern clinical radiation oncology. The text starts with an introduction to the basic physics principles, quantities, and radiation interactions as well as radiation production (chapters 1‐9). It is followed by descriptions of the properties of megavoltage photon and electron beams, treatment planning, and radiation measurements. The last section of the book focuses on advanced topics such as quality assurance, image guidance, and specific treatment techniques including SBRT/SRS, TBI, TSET, and particle therapy. This book not only covers an impressively broad range of medical physics topics, but is also up to date with modern advanced technologies in radiotherapy. For example, MR‐guided linac and MR‐guided radiation therapy are discussed in Chapter 9 and 21, respectively, which present the current frontiers in radiation therapy. Instead of providing a comprehensive bibliography, corresponding chapters in the classic textbooks such as Khan and McDermott are listed at the end of each chapter for further in‐depth reading. A set of 10 questions is provided for the readers to enhance their learning and understanding by solving problems related to each chapter. A total of 60 teaching videos are provided free on the publisher's website (3The strength of this book lies in its simplicity. It is written concisely with words that are intelligible to the readers who do not possess strong physics and mathematics backgrounds. The text is supplemented by numerical color illustrations to facilitate the learning of complicated physics concepts and processes. In addition, most pictures related to radiation treatment plans and dose distributions are from commercial treatment planning systems with actual patient images, making it easy to connect the knowledge to clinical practice.The most exceptional feature of this book is its interactive video series. As an instructor of a radiation physics course for clinical residents in a classroom setting for more than a decade, I found it extremely difficult to teach the course online this past year due to the social distancing requirement caused by the pandemic. To facilitate online classroom interaction and engagement, I started exploring the flipped classroom model by recording the lectures prior to class. However, it was very time consuming and also required extensive video editing skills. As a result, I was only able to flip a small portion of the lectures although they were well received by our residents. I was thrilled to find that this book provides a comprehensive series of teaching videos that can be very helpful in flipping the entire course into a more powerful interactive learning experience.4Primer on Radiation Oncology Physics by Eric Ford is an excellent textbook for diverse trainees in radiation oncology who take the first course in radiation therapy physics. It is also valuable to medical physics graduate students and residents as a refresher and review resource. The book is ideally suited for new teaching models with interactive videos and extensive problem sets, and it directly meets the educational needs of radiation oncology professionals with diverse backgrounds.In my opinion, Minsong Cao, Ph.D., DABR, FAAPM, is an Associate Professor and Associate Vice Chair of Education of the Department of Radiation Oncology, University of California, Los Angeles.