Due to degeneration of dopaminergic neurons and thus lower expected radioligand binding to DAT, higher measurement variability in PD patients might be expected than earlier reproducibility results in healthy controls. Therefore, we aimed to examine the test-retest properties of [18F]FE-PE2I-PET measurements within 7–28 days. The outcome measure was non-displaceable binding potential generated using wavelet-aided parametric imaging with cerebellum as reference region. We assessed test-retest performance using estimates of reliability and repeatability. Regions for primary analysis were caudate, putamen, ventral striatum, and substantia nigra. Exploratory analysis was performed for functional subdivisions of the striatum. We also compared the more vs. less affected side.Nine patients with PD (Hoehn and Yahr stage < 3) were included . Each patient underwent two [18F]FE-PE2I showed absolute variability estimates of 5.3–7.6% in striatal regions and 11% in substantia nigra and ICCs of 0.74–0.97 (median 0.91). The absolute variability for functional striatal subdivisions was 6.0–9.6% and ICCs of 0.76–0.91 (median 0.91). The less affected substantia nigra exhibited greater consistency than the more affected side. According to power calculations based on the current sample size, DAT changes of 5–11% in the striatum and 28% in the substantia nigra can be detected with a power of 0.8 (p < 0.0125).[18F]FE-PE2I in PD patients showed good repeatability and reliability. The slightly lower reliability in the substantia nigra in patients may be explained by lower DAT density and smaller anatomical size. Power calculations suggest that [18F]FE-PE2I PET is a suitable marker for longitudinal DAT decline in PD.DAT-PET measurements with [2017-003327-29EudraCT  Parkinson’s disease (PD) is a movement disorder characterized by progressive degeneration of the dopaminergic system, affecting both the cell bodies in the substantia nigra and its projections to the striatum. Within the dopaminergic system, the dopamine transporter regulates the synaptic dopamine levels by dopamine reuptake, and its density reflects presynaptic functioning.18F]FE-PE2I, developed in 2009 [18F]FE-PE2I in twenty patients with early-stage PD showed an in vivo striato-nigral gradient of DAT loss [18F]FE-PE2I-PET can be achieved with a shortened imaging protocol, making clinical implementation realistic [18F]FE-PE2I has been previously studied in twelve young, healthy men [18F]FE-PE2I as a disease progression marker, it is critical to assess reliability also in patient samples. Due to the degeneration of striatal projections, patients with PD have lower DAT availability, which could lead to lower measurement reliability compared with healthy controls.8FFE-PE2IDAT loss , 11. AftDAT loss , additioealistic , 12. Thelthy men , showingIn the majority of PET studies of the striatal dopaminergic system, the regions of interest are the striatum, divided into caudate, putamen, and ventral striatum (nucleus accumbens), and the substantia nigra (midbrain). This anatomical subdivision of the striatum, although useful, might not represent the functional organization of the striatum. Instead, subdivision based on the connectivity between the basal ganglia and the neocortex can be used, where the striatum is divided into a limbic, associative, and sensorimotor striatum  . The funThe assessment of the test-retest reliability of DAT-PET in patients with PD is relevant for several reasons. First, the knowledge on the natural variability is essential for interpretation of longitudinal follow-up study results; second, the measured variability can be used to estimate the minimum effect size on DAT needed for disease-modifying treatment trials; and third, for the purpose of power calculations for future clinical studies investigating longitudinal treatment efficacy.18F]FE-PE2I measurements in the main striatal areas and substantia nigra in patients with PD. The hypothesis was that the reliability in the striatum would be similar to that observed in healthy subjects, and the reliability in the substantia nigra lower than in the striatum considering the lower DAT density in the substantia nigra. The secondary objective was to evaluate the test-retest reliability of three connectivity-based functional subdivisions of the striatum in view of future PET analyses.The primary objective of this study was, therefore, to assess the test-retest reliability of FE-PE2I was prepared as previously described [18F]FE-PE2I PET measurements were acquired in each subject within an interval of 7–28 days  using 3D OP OSEM with 10 iterations and 16 subsets, including modeling of the PSF . Frame-tHead motion was evaluated by patient observation during data acquisition as well as during image analysis by reviewing the realignment plots and brain time activity curves (TACs). Translation of more than 3 mm on the realignment plots led to additional motion correction using an in-house developed automatic procedure. Description of the method is given in Supplementary Text http://surfer.nmr.mgh.harvard.edu/): whole striatum (STR), caudate (CAU), putamen (PUT), ventral striatum (VS), and cerebellum. For substantia nigra (SN), the functional molecular template, as created in the research group FE-PE2I were generated using wavelet-aided parametric imaging (WAPI) FE-PE2I measurement reproducibility was determined with calculation of repeatability  and reliability , as per recommendation of Weir, Baumgartner, and Matheson FE-PE2I can be used as DAT biomarker in PD.This study was designed to assess the test-retest reproducibility of [18F]FE-PE2I test-retest study in twelve young, healthy controls FE-PE2I FE-PE2I PET provides a comprehensive assessment of the PD pathophysiology in vivo.The higher resolution of PET compared with SPECT permits a better assessment of low-binding regions, such as the SN. Test-retest repeatability and reliability in this region were inferior to test-retest metrics in the striatum. The smaller size of the SN and the smaller numerical value of ]FE-PE2I  showed c18F]FE-PE2I PET is a reliable research tool. This is relevant for studies using the functional rather than anatomical striatal subdivisions in assessing correlations of DAT availability to specific clinical variables of cognitive function or behavior.The low variability and high reliability observed in the connectivity-based functional subdivisions of the striatum furthermore indicate that [18F]FE-PE2I PET is well suited for measuring biological DAT differences in striatal regions in a longitudinal follow-up study with a typical sample size for PET studies.Using the variability in test-retest differences within each region, we estimated the size of within-individual changes which could be detected with a power of 80%, using a significance threshold of 0.0125 . These estimates suggest that a study with a sample size of 9 patients is sufficiently sensitive to detect within-individual differences of greater than between 5 and 11% for different regions within the striatum and greater than 28% for the SN (Supplementary Table S18F]FE-PE2I measurements of DAT have good reliability in Parkinson patients (H&Y 1–2.5) even in the small anatomical areas with lower DAT density, such as the substantia nigra. The test-retest metrics were equal-to-superior to other DAT radioligands. Thus, this study further supports the suitability of [18F]FE-PE2I as imaging marker for longitudinal follow-up studies in PD.[Additional file 1: Supplementary material.


Coordinated activation of muscles is the basis for human locomotion. Impaired muscular activation is related to poor movement performance and disability. To restore movement performance, information about the subject's individual muscular activation is of high relevance. Surface electromyography (sEMG) allows the pain-free assessment of muscular activation and many ready-to-use technologies are available. They enable the usage of sEMG measurements in several applications. However, due to the fact that in most rehabilitation applications dynamic conditions are analyzed, the correct interpretation of sEMG signals remains difficult which hinders the spread of sEMG in clinical applications. From biomechanics it is well-known that the sEMG signal depends on muscle fiber length, contraction velocity, contraction type and on the muscle's biomechanical moment. In non-isometric conditions these biomechanical factors have to be considered when analyzing sEMG signals. Additionally, the central nervous system control strategies used to activate synergistic and antagonistic muscles have to be taken into consideration. These central nervous system activation strategies are rarely known in physiology and are hard to manage in pathology. In this perspective report we discuss how the consideration of biomechanical factors leads to more reliable information extraction from sEMG signals and how the limitations of sEMG can be overcome in dynamic conditions. This is a prerequisite if the use of sEMG in rehabilitation applications is to extend. Examples will be given showing how the integration of biomechanical knowledge into the interpretation of sEMG helps to identify the central nervous system activation strategies involved and leads to relevant clinical information. The coordinated activation of muscles forms the basis for human movement. A frequent consequence of lesions of the central nervous system is muscle paresis accompanied by reduced muscle force and/or the loss of the ability to activate the muscles in a coordinated way. This results in poor movement performance and causes pain and disability. To preserve and restore movement performance is a challenge, and the demand for more effective treatment methods is gaining more importance , 2. HoweSEMG technologies allow the pain-free assessment of muscular activation and many ready-to-use technologies are available , 5. TheyThis perspective report is addressed to clinical users but especially to sEMG developers. The aim is to raise awareness of the importance of biomechanical factors in the analysis of sEMG signals acquired under non-isometric conditions.One major barrier limiting the use of sEMG in clinical applications is that the correct interpretation of sEMG signals remains debatable , 7. ConsThe challenges sEMG is facing are related to the fact that most applications in rehabilitation are associated with non-isometric conditions. If information about the onset and cessation of muscular activation is to be extracted from the sEMG signal, unambiguous conclusions are possible as long as the sEMG signal can be related to individual movement phases. A particularly clinically relevant example for this approach is the determination of phases of muscular activation during gait . TherebyThe relationship between muscle force and sEMG signal has closely linked the disciplines of biomechanics and electromyography for decades , 13–17. These biomechanical factors affect the number of muscle fibers which must be excited to generate the force necessary to execute the movement. Since the sEMG amplitude depends on the number of excited muscle fibers, it is obvious that in non-isometric contractions sEMG amplitude varies with different biomechanical factors. In addition, agonistic, synergistic and antagonistic muscles generally act on a common joint and produce a resulting total net joint torque , 37–41. Normalization of the sEMG amplitude to force or torque is frequently used to counteract the high variability of the sEMG signal . It alloMany disorders that require rehabilitation are associated with the altered control and activation of muscles by the central nervous system and with the progressive development of rheological modifications in soft tissues, joint deviations and deformities that alter the biomechanics of the system and add mechanical boundaries . PromineIn order to be able to determine for each patient specifically, which individual alterations the activation strategy occur during the execution of movements, the physiological activation pattern of the muscles involved should be used as a baseline , 50. HowVon Werder et al. referred to the separation of the sEMG signal into near-isometric epochs as categorization . They usIn contrast to concentric contractions, the force that a single sarcomere can produce in eccentric contractions increases as the velocity of muscle stretch increases. Based on this biomechanical consideration, the sEMG envelope should decrease with increasing movement velocity during eccentric contractions. However, this can only be noticed in the biceps . The sEMParticularly in rehabilitation, there is a great demand to be able to evaluate functionality in everyday situations . This isSince, according to the definition of Lance, stretch velocity dependency is a characteristic property of spasticity, the question arises whether the integration of biomechanical knowledge might support a distinction between voluntary muscle activation and spasticity. As discussed before, to achieve comparability between physiological and spastic muscular activation patterns in dynamic conditions, near-isometric epochs of the sEMG signal have to be compared. As in healthy subjects, the biceps' sEMG envelope increases in patients with increasing contraction velocity during concentric contractions. This relationship is reflected by a positive gradient . HoweverThe sEMG signal is significantly dependent on various biomechanical factors and it can be assumed that an interpretation of the sEMG signal with respect to amplitude becomes more accurate when these biomechanical factors are taken into account. Hence, isometric measurements, in which these factors can be controlled, are still widely used. In rehabilitation, however, isometric contractions usually play a subordinate role. Here, the analysis of intentionally executed movement tasks is of primary importance. This complicates the interpretation of the sEMG signals, if beside the timing, the magnitude of the muscular activation is of interest. The consequences are often contradictory results both between individual studies and between follow-up measurements. This is fatal for the translation of the sEMG into clinical application, as it essentially weakens the users' confidence in the methodology.Among others, Bogey et al. emphasized the clinical relevance of an integrated analysis of timing and relative magnitude of the sEMG signal . ConsequWhen analyzing non-isometric conditions, additional measurement methods must be applied synchronously to the sEMG signal, which provide information about the execution of movements, such as movement cycle intervals, joint positions, movement velocities and external forces. This approach is already successfully applied in clinical gait analysis  and needOn the basis of current biomechanical knowledge, information about the execution of the movement must be merged with the sEMG signal. This will probably only be reliable if the analysis is broken down into near-isometric epochs. Appropriate algorithms that make this possible must be developed in the future.The two examples given show how the consequent implementation of this strategy leads to new insights important for rehabilitation. They give a representative of the potential of integrating biomechanical principles into the interpretation of the sEMG.In conclusion, there is an increasing demand in rehabilitation for objective methods, which on one hand provide evidence and on the other hand enable a therapy tailored more to the individual patient. sEMG has the potential to contribute significantly to this goal, even in dynamic conditions . HoweverThe data analyzed in this study is subject to the following licenses/restrictions: The copyright for the data belongs to the Department of Rehabilitation and Prevention Engineering; Institute of Applied Medical Engineering; RWTH Aachen. Requests to access these datasets should be directed to Catherine Disselhorst-Klug, disselhorst-klug@ame.rwth-aachen.de.The studies involving human participants were reviewed and approved by Ethik-Kommission der Medizinischen Fakultät an der RWTH Aachen. The patients/participants provided their written informed consent to participate in this study.CD-K elaborated the relationship between the surface EMG and the biomechanical factors, identified the barriers for translation into clinical application, developed the measures necessary to overcome them, reinterpreted the data from the presented studies, and wrote the manuscript. SW conducted an extensive literature research on the context of the manuscript, involved in the reinterpretation of the study data, assigned the results to the state of the art, and revised the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Preclinical studies suggest that interactions between granulocyte colony-stimulating factor (G-CSF) and hypoxia-induced carbonic anhydrase IX regulate the trafficking and function of immune cells in the tumour microenvironment. We investigated the clinical significance of this crosstalk by analyzing the protein expression of G-CSF and macrophage markers by immunohistochemistry on a well-characterized tissue microarray series of invasive breast cancers. We report that high expression of G-CSF on breast carcinoma cells is linked with significantly improved survival in an important group of breast cancers that do not respond to hormonal therapy. These tumours were infiltrated by immune cells expressing biomarkers that can be targeted with immune checkpoint inhibitor drugs. In contrast, carbonic anhydrase IX expression was associated with unfavourable outcomes.high expression showed a significant positive association with ER negativity, HER2 positivity, presence of CD163+ M2 macrophages, and CAIX expression. In univariate analysis, G-CSFhigh phenotype was associated with improved survival in non-luminal cases, although the CAIX+ subset had a significantly adverse prognosis. A significant positive association was observed between immune checkpoint biomarkers on tumour-infiltrating lymphocytes and both G-CSF- and CAIX-expressing carcinoma cells. Immune checkpoint biomarkers correlated significantly with favourable prognosis in G-CSFhigh/non-luminal cases independent of standard clinicopathological features. Conclusions: The prognostic associations linking G-CSF to immune biomarkers and CAIX strongly support their immunomodulatory roles in the tumour microenvironment.Purpose: Granulocyte colony-stimulating factor (G-CSF) and hypoxia modulate the tumour immune microenvironment. In model systems, hypoxia-induced carbonic anhydrase IX (CAIX) has been associated with G-CSF and immune responses, including M2 polarization of macrophages. We investigated whether these associations exist in human breast cancer specimens, their relation to breast cancer subtypes, and clinical outcome. Methods: Using validated protocols and prespecified scoring methodology, G-CSF expression on carcinoma cells and CD163 expression on tumour-associated macrophages were assayed by immunohistochemistry and applied to a tissue microarray series of 2960 primary excision specimens linked to clinicopathologic, biomarker, and outcome data. Results: G-CSF Granulocyte colony-stimulating factor (G-CSF) is a member of the colony-stimulating factor superfamily and exerts its biological effects by binding to the G-CSF receptor (G-CSFR). The archetypal G-CSF/G-CSFR signalling pathway is critical for proliferation and survival of myeloid precursors and their subsequent differentiation into neutrophils with augmentation of their effector functions . Recogni2 into bicarbonate ions and protons. The bicarbonate is transported into the cells via bicarbonate transporters, thereby aiding in the buffering of intracellular pH, whereas the protons remain in the extracellular space, contributing to extracellular acidification [Tumour hypoxia is a hallmark of invasive tumours and is associated with genomic instability, the emergence of therapy-resistant clones, inhibition of anti-tumour immune responses, and cancer progression ,17. Carbfication . We and fication ,20,21. Ification . Furtherfication . Thus, tfication ,24,25,26Tumour-associated macrophages comprise heterogeneous populations of cells that exhibit remarkable plasticity and play an important role in modulating adaptive immune responses . HoweverThus, existing data from preclinical studies highlight hypoxia and inflammation as critical modulators of the immune microenvironment of solid tumours where a significant interplay between G-CSF and CAIX has been found to play a role. Presently, it is not known whether there is an association amongst G-CSF, CAIX, and immune biomarkers in breast cancer clinical samples. Furthermore, to date, only limited studies  have evaluated the prognostic significance of G-CSF expression in breast cancer . Hence, n = 330). The clinicopathological characteristics, adjuvant treatments, and inclusion/exclusion criteria have been described previously [n = 2960) of female patients diagnosed with stage I–III breast cancer in the province of British Columbia at the British Columbia Cancer Agency between 1986–1992, for which formalin-fixed paraffin-embedded tumour blocks were accessible. The adjuvant systemic and endocrine therapies recommended in the specified years were in accordance with the provincial standards. No patients received immune checkpoint inhibitor therapy. The clinicopathological characteristics of this cohort have been formerly described [We first established the immunohistochemical (IHC) staining procedure, scoring methodology, and interpretation of G-CSF protein expression on a series of female breast cancer patients diagnosed with stage I–III disease during 1998–2002  guidelines .The construction of the tissue microarrays used in this study has been described previously ,38,39. IThe IHC protocols are detailed in http://www.gpec.ubc.ca/gcsf accessed on 1 January 2021), and representative images are shown in The scoring of G-CSF expression was adapted from Hollmen et al. . The int® SPSS software (version 25) and R (v 3.3.2) were used for statistical analyses. Relevant descriptive statistics were computed for continuous and categorical variables. The correlation of G-CSF with clinicopathological factors and key biomarkers was assessed by chi-square. The primary endpoint for the clinical outcome was breast cancer-specific survival (BCSS), specified as the time interval between the date of diagnosis and the date of breast cancer-associated mortality. Patients who were alive at the end of follow-up or died due to non-breast cancer-related causes were censored. Overall survival (OS) and relapse-free survival (RFS) were used as secondary endpoints. OS was defined as the time period from the date of diagnosis till the date of last follow-up or death due to any cause, whereas RFS was defined as the time period from the date of diagnosis until any breast cancer-related relapse, whether local, regional, distant, or contralateral. Univariate analysis for survival probabilities was computed by the Kaplan–Meier method, and differences in the survival rates between analyzed groups were estimated by log-rank test. Cox proportional hazard modelling was used for multivariable analysis, and adjusted hazard ratios with 95% confidence intervals were reported for each variable included in the model. A p-value of <0.05 was considered statistically significant. To address the concerns of multiple comparisons while assessing associations between G-CSF expression and clinicopathological features, a Bonferroni correction was applied, making the criterion for statistical significance p < 0.003.IBMCohort I was used to evaluate the performance of G-CSF immunostaining, finalize the scoring methodology, and explore cut points such that these could be locked down prior to assessment of the main study cohort II. The correlative analyses with clinicopathological features and association with primary and secondary endpoints for cohort I are presented in n = 2960), data from which has been previously published for expression of immune biomarkers and CAIX [We describe hereafter the results of our detailed analysis on cohort II , and the median duration of follow-up was 12.5 years (range: 0.1–18.5 years). A total of 1956 deaths were recorded in the entire cohort, of which 58.4% were attributed to breast cancer. The clinicopathological characteristics of the study cohort are summarized in low cases that comprised 53.3% of the study population  negativity, HER2 positivity, CD163+ M2 macrophages, CAIX expression, and (IHC-defined) HER2 and basal intrinsic breast cancer subtypes, relative to the expression of these biomarkers among the G-CSFpulation .p = 0.43) nor when the analysis was restricted to the luminal subtype, which comprised 74% of the G-CSF interpretable cases . In contrast, amongst the non-luminal subtype, high expression of G-CSF correlated with a significantly improved BCSS   . Similar = 0.02) .n = 2960) using Cox proportional hazards model for multivariate analysis . However, within the non-luminal cases (n = 665), G-CSFhigh tumours correlated with a 28% reduced risk of breast cancer-related deaths compared to G-CSFlow tumours, independent of the standard clinicopathological factors including age, tumour size, grade, lymphovascular invasion, and axillary lymph node metastasis  (high expression was significantly associated with better BCSS in HER2 subtype (n = 196); triple-negative subgroup (n = 469) and basal (n = 274) subgroups of non-luminal breast cancers did not generate hazard ratios that remained significant in multivariable analysis (data not shown).Consistent with the univariate analysis, G-CSF expression was not an independent prognostic indicator in the whole cohort ( = 0.01) . Within high phenotype, significantly adverse BCSS  was observed in CAIX-expressing tumours , with a similar favourable trend observed for breast cancer-specific and overall survival , lymphocyte activation gene-3 (LAG-3), and T-cell immunoglobulin domain and mucin domain-3 (TIM3)) or downregulate normal T-cell responses by increasing FOXP3 regulatory T cells . To addrrophages .Considering the immune-modulating role of G-CSF, we performed a multivariable analysis to investigate the individual prognostic significance of immune checkpoint biomarkers in G-CSF-expressing non-luminal cases. We found that intratumoural lymphocytes expressing CD8, PD-1, FOXP3, TIM3, and LAG3, as well as PD-L1-expressing carcinoma cells, were each associated with better survival, independent from standard clinicopathologic factors . SimilarTaken together, our results demonstrate that the prognostic value of G-CSF in non-luminal breast cancers is influenced by tumour microenvironmental features associated with CAIX positivity.We herein report the prognostic significance of G-CSF expression on breast carcinoma cells in a large population-based cohort of stage I–III invasive breast cancers. The most salient findings of our study were (a) a survival advantage associated with high expression of G-CSF in non-luminal subtypes of breast cancer; (b) a novel association between G-CSF, CAIX, and markers of immune exhaustion on tumour-infiltrating lymphocytes, which is consistent with the presence of an immune-suppressive hypoxic and acidic tumour microenvironment; (c) the identification of a patient subset among non-luminal breast cancers with high G-CSF and CAIX expression that is associated with worse survival compared to those that did not express CAIX; and (d) a positive correlation between tumour-infiltrating lymphocytes expressing immune checkpoint biomarkers and carcinoma cells expressing G-CSF, such that their concurrent presence is associated with a survival advantage in non-luminal immune-enriched subtypes.The G-CSF/G-CSFR signalling axis has been implicated in stimulating neoangiogenesis, tumour cell proliferation, enhanced migratory and metastatic potential, and expansion of the cancer stem cell pool in preclinical models of several solid organ malignancies . AdditioFOXP3 transcriptional upregulation enriches for FOXP3+ T regulatory cells, which play a crucial role in self-tolerance [high and CD163+ M2 macrophage-infiltrated non-luminal breast cancers.It is well documented that tumour hypoxia and ensuing acidification of the microenvironment promote immunological escape and resistance to immunotherapy by several mechanisms ,54. Firsolerance . Thirdlyolerance . Moreoveolerance . Consistolerance . Our datThe identification of a soluble marker, G-CSF, whose expression is associated with the expression of CAIX in a clinical cohort, is an important finding. Our preclinical findings linked CAIX and G-CSF through the NF-κB signalling pathway . Non-lumCA9, which encodes CAIX [Differences between the prognostic significance of G-CSF in our study and those previously reported in other tumour types may be attributed to fundamental differences in the underlying tumour biology. For example, inactivation of tumour suppressor pVHL is observed in up to 80% of clear cell renal cell carcinomas, yet this is rare in breast carcinomas ,70. The des CAIX . Similardes CAIX ,72 of thdes CAIX . Hence, n = 127) with high G-CSF expression were associated with poor overall survival [To date, limited studies have investigated the significance of G-CSF protein expression in breast cancer. Considering the strong association with neutrophilic mobilization and recruitment in carcinomas, serum G-CSF has been evaluated as a surrogate biomarker for prognostication. In this context, a few studies have shown a higher plasma level of G-CSF in breast cancer patients compared to healthy controls and post-surgical wound-healing fluids ,74,75. Ssurvival . Though CSF3R was identified as one of the differentially expressed genes in a subset of immune-rich ER+ breast cancers [n = 284), and we observed no significant difference in survival between G-CSFhigh and G-CSFlow tumours (data not shown).In a recent study, using publicly available data sets,  cancers . AlthougOur study has major strengths, including the use of analytically validated antibodies, predefined scoring criteria (CD163 and CAIX), and the use of an independent cohort (cohort I) to confirm the performance of G-CSF as a biomarker on breast cancer tissue microarrays. However, there are some important limitations worthy of mention. First, we had hoped to see significant findings in cohort I  that could guide prespecified formal hypotheses testing using cohort II as an independent validation set. However, the results on the training set were not significant for our primary endpoint in univariable analysis. Instead, we were able to use the findings from cohort I to lock down the staining methodology, interpretation, and cutpoints, but our observations using the more powered cohort II with longer-term follow-up will need independent validation. Secondly, we did not evaluate the presence of tumour-associated neutrophils or myeloid-derived suppressor cells, which are the two major immune cell types mobilized and recruited to the tumour site by G-CSF. Instead, we took into consideration the expression of immune checkpoints on tumour-infiltrating lymphocytes as a related biomarker, hypothesizing that G-CSF mobilizes immune infiltrates and promotes an immune exhaustion phenotype on tumour-infiltrating lymphocytes. Thirdly, we limited our analysis to G-CSF expression by the carcinoma cells, and these results do not take into account the expression of G-CSF on immune cells, such as macrophages, endothelial cells, or other stromal cells that are part of the tumour immune microenvironment. The use of tissue microarrays may be considered a potential limitation of our study. While there are more than a few studies showing considerable agreement between cores derived from source tumour blocks and the full-face sections ,78, thisIn conclusion, our results demonstrate the prognostic significance of G-CSF in invasive breast cancer, whereby high expression serves as an indicator of better survival in aggressive non-luminal subtypes of breast cancer in the absence of CAIX. Our identified associations between CAIX, G-CSF, and immune biomarkers provide a rationale for additional prospective investigations to understand the underlying mechanisms and their role as potential biomarkers for predicting responses to immune checkpoint inhibitor therapy.


Albumin, the most abundant plasma protein, not only controls osmotic blood pressure, but also serves as a carrier for various small molecules, including pharmaceuticals. Its impact on pharmacological properties of many drugs has been extensively studied over decades. Here, we focus on its interaction with the following mobilizing agents: Granulocyte-colony stimulating factor (G-CSF) and AMD3100, where such analyses are lacking. These compounds are widely used for hematopoietic stem cell mobilization of healthy donors or patients. Using albumin-deficient (Alb−/−) mice, we studied the contribution of albumin to mobilization outcomes. Mobilization with the bicyclam CXCR4 antagonist AMD3100 was attenuated in Alb−/− mice compared to wild-type littermates. By contrast, mobilization with recombinant human G-CSF (rhG-CSF), administered twice daily over a five-day course, was significantly increased in Alb−/− mice. In terms of a mechanism, we show that rhG-CSF bioavailability in the bone marrow is significantly improved in Alb−/− mice, compared to wild-type (WT) littermates, where rhG-CSF levels dramatically drop within a few hours of the injection. These observations likely explain the favorable mobilization outcomes with split-dose versus single-dose administration of rhG-CSF to healthy donors. Albumin is, by a wide margin, the most abundant plasma protein of all higher species, including man, typically at concentrations of 30–50 g/L. It not only controls colloid osmotic pressure ,2,3 but Hematopoiesis is the continuous, life-long process of generating short-lived mature blood cells that originate from hematopoietic stem and progenitor cells (HSPC). The process of proliferation and cell differentiation is tightly controlled. Individual HSPC will undergo asymmetric cell divisions, whereby progenitors arise. Under the influence of their environment in the bone marrow (BM), they will give rise to increasingly more differentiated precursors and mature blood cells, which are ultimately released into the circulation . The neaOur interest in HSPC is significantly driven by translational aspects. In particular, the enforced egress of HSPC, so called mobilization, for graft manufacturing, remains a focus of our studies. HSPC are routinely collected from the peripheral blood of healthy donors after a 5-day course of mobilization with recombinant human granulocyte-colony stimulating factor (rhG-CSF) ,15. As aSerum albumin plays a crucial role in the transport of small molecules and drugs through the blood stream, and as such is a potential carrier of both AMD3100 and rhG-CSF. As was shown, up to 58% of administered AMD3100 directly binds to plasma proteins [em8Mvw/MvwJ (Alb−/−) mice were described previously [For all experiments, young adult mice  were used. C57BL/6J-Albeviously , albeit Peripheral blood was drawn from the facial vein into EDTA tubes using a 23G needle. Total cell counts were analyzed by Hemavet 950SF+ . BM cells were harvested by aseptically flushing femurs and/or tibiae using Phosphate-Buffered Saline (PBS) + 0.5% Bovine Serum Albumin (BSA). Splenic cells were obtained by aseptic blunt extrusion of the capsule.Hematopoietic cells in blood, BM, and spleen were enumerated using multi-parametric flow cytometry for informative markers, for mature and immature subsets, as well as in vitro clonogenic assays ,33,34. AProgenitor cells were mobilized into peripheral blood using either AMD3100  or rhG-CSF  either in 0.9% NaCl  or 20% human albumin solution . Blood was drawn 1 h, 2 h, and 4 h after administration of AMD3100 or 1 h after the last administration of rhG-CSF (unless otherwise stated), followed by colony-forming unit culture (CFU-C) enumeration in blood, as well as, where indicated, in BM and spleen. WBC were counted at every time point. LSK, LSK-SLAM, and cell cycle analysis were performed in BM and spleen cell suspensions using flow cytometry.Human serum albumin in mouse serum was detected and quantified using human albumin ELISA  according to the manufacturer’s recommendations.Human G-CSF Flex Set  was used to quantify levels of rhG-CSF in murine plasma or BM-fluid samples.t-tests were calculated using Excel ; twoway ANOVA with Bonferroni post-test was calculated using GraphPad Prism 5 ; for non-normally distributed data, the nonparametric Mann–Whitney U test was calculated using SPSS . Unless stated otherwise, results are presented as mean ± standard error of the mean (SEM), ns p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001.Descriptive statistics and students’ In order to assess the effect of albumin deficiency on hematopoietic stem cell mobilization, we first enumerated phenotypically and functionally mature and immature hematopoietic cells in all hematopoietic organs of untreated young adult Alb−/− mice, or wild-type littermates as baseline values. The numbers of mature leukocytes in all compartments  were normal in Alb−/− mice . MoreoveMice received a single i.p. bolus injection of AMD3100. WBC egress and accumulation in peripheral blood could be detected as early as 1 h after administration, with a maximum at 2 h in both Alb−/− and WT mice A irrespeSimilarly, modeling clinical mobilization with the more slowly acting cytokine G-CSF, mice received a total of nine, 12-hourly i.p. injections of rhG-CSF, followed by mature and immature leukocyte enumeration in blood, BM, and spleen. Interestingly, albumin-deficient mice showed a 2-fold higher WBC and HSPC mobilization into the circulation, when compared to WT controls , but theWe hypothesized that local rhG-CSF concentrations might be affected by albumin, and by that mechanism, lead to differences in mobilization efficiency. To test that, we studied the pharmacokinetics of rhG-CSF in plasma and BM fluids of Alb−/− and WT mice. In plasma, we observed the same high (15 ng/mL) and only slowly (over 8 h) decreasing levels of rhG-CSF in both Alb−/− and WT mice F. On theTo ascertain that the observed mobilization phenotype of the Alb−/− mouse is directly attributable to lack of albumin, as opposed to some complex compensatory mechanism, we also tested rhG-CSF in hAlb-substituted mice of both genotypes, Alb−/− and WT. Bioavailability of hAlb was independent of genotype, so that hAlb substitution raised albumin in Alb−/− mice to near-normal levels A. EndogeAlbumin is the most abundant serum protein, controlling osmotic pressure and acting as a cargo carrier ,2,3. In In the course of this study, we have addressed the impact of serum albumin on homeostatic hematopoiesis and pharmacological mobilization of HSPC. We have shown that homeostatic mature and immature hematopoiesis is independent from albumin. However, when it comes to drug delivery and drug efficacy of clinically relevant mobilizing agents, albumin has marked impact on hematopoietic outcomes, as similarly already shown for many other pharmaceuticals ,47,48. H


Delia root maggot species are commonly found in onions. It is unclear which species affects onions most and how they are distributed among the major onion growing regions in Eastern Canada. Through oviposition and feeding preference bioassays, we determined that all species can similarly affect onion crops. We also determined that proportions of Delia species differ between the growing regions.Delia antiqua, Delia platura and Delia florilega are three root maggot species commonly considered pests in Eastern Canadian onions. The onion maggot, D. antiqua, is considered the primary root maggot pest in onion but it remains unclear whether the other two species are also causing damage. In order to develop updated management strategies for root maggot, we tested adult oviposition and feeding preference by Delia larvae on four growth stages of onion using bioassays and we determined the Delia species composition across the four major onion growing regions in eastern Canada. Delia species oviposit readily on onion at the 5–7 true leaf growth stage but damage on onions is not statistically different between Delia species in our zero-inflated models. The four eastern Canadian onion growing regions have different proportions of Delia species. Southern Ontario and Quebec were the only two regions where Delia antiqua was collected. The highest average numbers of Delia spp. were caught in Quebec and Nova Scotia. Our study shows that timing is important in implementation of management strategies for root maggot in Eastern Canadian onions. Delia Robideau-Desvoidy (Diptera: Anthomyiidae) [Delia radicum (Linnaeus), the cabbage maggot, and Delia antiqua (Meigen), the onion maggot. Delia radicum feeds on members of the Brassiceae [D. antiqua feeds on various Allium species. Other species within the genus, e.g., Delia platura (Meigen), the seedcorn maggot, and Delia florilega (Zetterstedt), the bean seed maggot, are considered generalists. Delia platura can develop on over 40 host plants in multiple families [Delia florilega is polyphagous with the ability to feed and develop on legumes, garden crops and cereals [Canadian field vegetable and field crops can be host to several damaging root maggot species in the genus myiidae) . The twoassiceae ,3 while families . Delia f cereals .Delia species are commonly associated with onion crops: D. antiqua, D. platura and D. florilega [Delia spp. damage plants by feeding on the germinating seeds, seedlings and, if conditions are right, can even mine the stems, rendering the plants unmarketable. Losses of untreated onion plants attributed to D. antiqua feeding can average 24–40% in Canada, depending upon variety [D. antiqua is considered the primary pest of onions, studies have shown that D. platura and D. florilega may be causing a larger proportion of damage to onion bulbs than previously thought [D. platura and D. florilega may be primary invaders in onion [Three lorilega . Larvae  variety . Althoug thought ,8,9,10. in onion ,11.Delia maggots. Insecticides  are commonly applied at seeding to control the first generation of root maggots in the spring; however, D. antiqua has been showing signs of resistance to chlorpyrifos [Delia whether they attack onions or not. Cultural controls such as appropriate crop rotation and timed planting potentially control root maggot populations [Delia pest pressure [Current management strategies targeting root maggots in eastern Canadian onion crops include chemical, cultural and biological controls, but are applied non-discriminately to all species of rpyrifos ,12. Thisulations , howeverulations ,15,16 mapressure .Delia species in eastern Canadian onion growing regions . The combination of these will hopefully provide insight into management strategies for root maggots in onion.Even with multiple Integrated Pest Management (IPM) tools available to control root maggots, the probability of success is affected by the lack of knowledge about the species that are causing damage to the crops. The main objectives of our research were to use bioassays to (a) elucidate female preference to lay eggs at particular plant growth stages (hereafter oviposition), (b) determine feeding preference to identify which species actively damages onion and at what growth stage and (c) determine the diversity and distribution of Delia antiqua and D. platura puparia were obtained from London Research and Development Centre  and Montmorency College , respectively. Delia florilega was not included into the bioassays because we could not obtain or establish a laboratory colony for that species. Delia species were reared for one generation on an artificial diet as described in Ishikawa et al. [a et al.  and, fola et al. , maintaiAllium cepa cv.  was obtained from Stokes Seed, Thorold, Ontario, and sowed in plastic pots (4 cm diameter) with a standard greenhouse potting mix, watered as needed and maintained in the same growth chamber with the Delia colonies. When plants reached one of 4 different plant growth stages: (1) 5 days post seeding, (2) radical/flag leaf stage, (3) second true leaf stage (TLS) and (4) 5–7th true leaf stage (TLS), they were used in bioassays. The plant stages 2 through 4 lasted on average four days each.Onion seed of For each replicate, one mating pair (approximately 48–96 h old) was introduced into a BugDorm-1  with one onion plant for 48 h with 20 replicates for each growth stage and species combination. We chose 48–96 h old mating couples because that length of time allowed us to be certain that the females were mated and that they have not started laying yet. The plant was then destructively examined under a microscope for the number and placement of eggs .D. antiqua or D. platura larvae, which had hatched 24–36 h prior to the assay, at the specific plant growth stages, with 20 replicates for each density and root maggot species. Larvae remained in pots until pupae or adult flies were recovered. Once the plants reached the bulb stage, they were assessed for damage  using Akaike Criterion (AIC) (ZIP AIC score 387.49 and ZINB AIC score 326.21).The data for the oviposition bioassay was over-dispersed (variance = 57.16 > mean = 3.61) with an excess of zeros (76.3% zero counts) so we fitted a Zero-Inflated Generalized Linear Model with Negative Binomial distribution (ZINB) with plant growth stage and Delia species as explanatory variables and mean damage rating as the response. It was the best-fit model when compared with a ZINB using AIC (ZIP AIC score 1031.18 vs. ZINB AIC score 1033.18).Similarly, for the damage bioassays, the data were over-dispersed (variance = 4.75 > mean = 1.89) with an excess of zeros (46.9% zero counts). We fitted a ZIP Model with plant growth, larval density and All statistics were done with the pscl package ,21 in RSDelia species flying through the field. We used blue sticky cards because that is the color of sticky card used by Canadian growers to monitor for onion maggot. These sticky cards were changed twice a weekly from planting (late May) through to harvest (late July) in 2017  based on Bray–Curtis distance matrix was used to determine significant differences among Delia species assemblages by region using the function Adonis. All statistics were run with the vegan R package [For each region, the data were pooled over the entire season since there was little difference in diversity of  package  in RStud package .p < 0.01) showed a significant difference with respect to ovipositional preference in the zero-inflation portion of the models  and 5–7 TL growth stage . A single D. antiqua caused onion to be non-marketable (mean damage rating of 3) when fed upon at the 5–7 TL stage. With 3 larvae, the damage rating increased to 4 . The Mantel test showed that spatial distance between the regions is not auto correlated with Delia diversity . Delia species composition is more similar within regions then between regions  monitoring for Delia spp. should focus early in the planting season to determine if management of the crop is necessary before the vulnerable growth stage (5–7 TL), (2) monitoring and pulling non-healthy young onion plants to discourage Delia spp. from ovipositing and populations growing and (3) managing the field with chemical or biocontrol should focus when the plants are near a 5 TL stage if the Delia spp. populations are high. In order to prevent damage in onions by Delia spp. root maggots, biological control methods [Through these findings, we suggest a three-fold approach to managing  methods ,14,15 sh


Coronary microembolization (CME) has a poor prognosis, with ventricular arrhythmia being the most serious consequence. Understanding the underlying mechanisms could improve its management. We investigated the effects of granulocyte colony-stimulating factor (G-CSF) on connexin-43 (Cx43) expression and ventricular arrhythmia susceptibility after CME.n = 10 each): Sham, CME, G-CSF, and AG490 (a JAK2 selective inhibitor). Rabbits in the CME, G-CSF, and AG490 groups underwent left anterior descending (LAD) artery catheterization and CME. Animals in the G-CSF and AG490 groups received intraperitoneal injection of G-CSF and G-CSF + AG490, respectively. The ventricular structure was assessed by echocardiography. Ventricular electrical properties were analyzed using cardiac electrophysiology. The myocardial interstitial collagen content and morphologic characteristics were evaluated using Masson and hematoxylin-eosin staining, respectively.Forty male rabbits were randomized into four groups (P < 0.01), indicating less severe myocardial damage and arrhythmias. The G-CSF group showed higher phosphorylated-Cx43 expression (P < 0.01 vs. CME). Those G-CSF-induced changes were reversed by A490, indicating the involvement of JAK2. G-CSFR, phosphorylated-JAK2, and phosphorylated-STAT3 protein levels were higher in the G-CSF group than in the AG490 (P < 0.01) and Sham (P < 0.05) groups.Western blot and immunohistochemistry were employed to analyze the expressions of Cx43, G-CSF receptor (G-CSFR), JAK2, and STAT3. The ventricular effective refractory period (VERP), VERP dispersion, and inducibility and lethality of ventricular tachycardia/fibrillation were lower in the G-CSF than in the CME group (G-CSF might attenuate myocardial remodeling via JAK2-STAT3 signaling and thereby reduce ventricular arrhythmia susceptibility after CME. Early percutaneous coronary intervention (PCI) can restore blood flow to ischemic myocardium, decrease infarct size, and reduce mortality and complications , but aboConnexin-43 (Cx43) is the major protein of gap junctions in the cardiac ventricles and is crucial for the synchronized contraction of the heart . RemodelGranulocyte colony-stimulating factor (G-CSF) can reduce myocardial apoptosis and inflammation, inhibit ventricular remodeling, improve cardiac function, stabilize the myocardial electrophysiological characteristics, and reduce the incidence of ventricular arrhythmia after ischemia-reperfusion injury and MI . G-CSF cWe hypothesized that G-CSF would attenuate the structural and electrical remodeling of the myocardium after CME and reduce the susceptibility to ventricular arrhythmia by regulating Cx43 phosphorylation and distribution via the JAK2-STAT3 pathway. Therefore, we investigated the effects of G-CSF and a JAK2-STAT3 inhibitor (AG490) on myocardial remodeling and arrhythmia susceptibility in a novel rabbit model of CME.n = 10 per group). All experimental procedures were approved by the National Laboratory Animal Management Regulations and Fujian Provincial Laboratory Animal Management Regulations.Forty male New Zealand white rabbits (3.0–3.5 kg) were purchased from Songlian Laboratory Animal Center, Shanghai, China (certificate no. 2007001103447). All animals were housed at a constant temperature of 24 °C with a humidity of ~ 50% with a 12-h/12-h light/dark cycle , with food and water available ad libitum. After 1 week of adaptive feeding, the rabbits were randomly divided into four groups: CME, G-CSF, AG490, and Sham (− 1 im), intubated, and ventilated with supplemental oxygen (2–4%). During the procedure, the rabbits were anesthetized with continuous intravenous infusion of ketamine and xylazine (5 and 4.5 mg·kg− 1·h− 1). The right common carotid artery was exposed and punctured, a guidewire was inserted into the ascending aorta under X-ray fluoroscopy, and a catheter was introduced into the left coronary artery for non-selective coronary angiography. For rabbits in the CME, G-CSF, and AG490 groups, autologous thrombus particles  were injected via the catheter into the LAD artery to establish the CME model. For rabbits in the Sham group, an equal volume of normal saline was injected. The rabbits in the G-CSF group received subcutaneous injections of G-CSF (10 μg/kg/d) beginning at 2 h after surgery and then daily for 6 days [Each rabbit was sedated with ketamine and xylazine . Echocardiography was carried out using a Vivid 7 ultrasound instrument  and an S12 transducer. The following parameters were recorded: parasternal left ventricular end-diastolic dimension , left ventricular end-systolic dimension , interventricular septum dimension , left ventricle posterior wall dimension , fractional shortening , and left ventricular ejection fraction . The average of three independent measurements was used for the analysis.Echocardiography was performed before and 2 weeks after surgery, as previously described . Each ra− 1 im) and buprenorphine (0.03 mg·kg− 1 sq), intubated, and ventilated with isoflurane . Heart rate, QRS interval, and QT interval (lead II) were recorded using a multi-channel electrophysiology recorder. The right jugular vein and carotid artery were identified and separated and heparinized. Then, 4-French, quad-polar electrode catheters were inserted under X-ray guidance into the right and left ventricles through the jugular vein and carotid artery, respectively. The catheters were connected to a multi-channel electrophysiology recorder, and stimuli were applied at twice the diastolic threshold with a pulse width of 2 ms. The evaluated parameters were ventricular effective refractory period (VERP), VERP dispersion, and occurrence of ventricular tachycardia (VT) or ventricular fibrillation (VF). VERP was defined as the shortest S1-S2 interval that elicited an electrogram response in the absence of absolute refractoriness. VERP dispersion was defined as the maximum difference between VERPs measured at the left and right ventricular apex as well as the mitral valve and tricuspid annulus. The presence/absence of VT/VF was determined after the application of S1S1 stimuli (40-ms duration) at the right ventricular apex for 10 s, repeated three times at intervals of 3 min.The electrophysiology study was performed using a modification of a method described previously . Two wee− 1, i.v.), and the heart was excised and washed in 0.9% pre-cooled (4 °C) saline. A portion of the left ventricle containing the papillary muscles was fixed in 10% neutral formalin for 24 h, embedded in paraffin, and sectioned to produce 10 consecutive sections with a thickness of 3.5 μm.After completion of the electrophysiology experiments, the animals were sacrificed with an overdose of pentobarbitone sodium . The ratio of the microinfarcted area to the total area (%) was calculated using the Image-Pro Plus (IPP) 6.0  image analysis software.Standard techniques were used to stain paraffin-embedded sections with Wiegert’s iron hematoxylin (10 min), plasma stain , phosphomolybdic acid (5 min), and aniline blue (15 min). Collagen fibers are stained blue, cardiomyocytes are stained red, and nuclei are stained blue-black. Sections were observed under the DMI3000B microscope. IPP6.0 was used to calculate the ratio of the area of collagen in the ventricular interstitium (excluding collagen around blood vessels) to the total area.The expression levels of phosphorylated Cx43 (p-Cx43), total Cx43 (t-Cx43), and G-CSFR in the anterior ventricular wall were evaluated. The paraffin sections were dewaxed, blocked with goat serum, and incubated overnight at 4 °C with goat anti-p-Cx43 , anti-t-Cx43 , or anti-G-CSFR  primary antibody. The sections were incubated with mouse anti-goat secondary antibody labeled with horseradish peroxidase (HRP)  for 20 min at 37 °C. The sections were developed using streptavidin and diaminobenzidine. Negative controls were incubated with phosphate-buffered saline instead of antibodies. After mounting with resinene, the sections were evaluated under high magnification (100× and 400×).Protein samples were extracted from left ventricular myocardium using Protein Extraction Kit . The protein concentrations were determined using the bicinchoninic acid protein assay . Equal amounts (50 μg) of protein were subjected to 10% SDS-PAGE, transferred to a polyvinylidene difluoride membrane, blocked with 5% non-fat milk in Tris-buffered saline , and incubated overnight at 4 °C with a primary antibody: anti-p-Cx43 , anti-t-Cx43 , anti-G-CSFR , anti-t-JAK2 , anti-p-JAK2 , anti-t-STAT3 , anti-p-STAT3 , or anti-β-actin . β-actin was used as internal control. After three washes in TBS-Tween-20, the membranes were incubated with the appropriate secondary antibody labeled with HRP  in the dark for 1 h at room temperature. Protein bands were developed by electrochemiluminescence, and their intensities were analyzed using the ImageJ software .P < 0.05 was considered statistically significant. Prism 5  was used for graph plotting.Data are presented as means ± standard deviations and were analyzed using SPSS 19.0 . Normally distributed data (according to the Kolmogorov-Smirnoff test) with homogeneity of variance were compared between groups using one-way analysis of variance (ANOVA) and the least significant difference (LSD) test. Data within each group between baseline and 2 weeks were tested using the paired t-test. Data that were not normally distributed or had heterogeneity of variance were analyzed using the Kruskal-Wallis U test. VT/VF rate and mortality rate were compared using the chi-square test. P < 0.01), G-CSF (P < 0.01), and AG490 (P < 0.05) groups  and a significantly lower LVEF and FS (P < 0.01) than the Sham and G-CSF groups  and significantly shorter in the AG490 group than in the Sham group (P < 0.01) , with no significant differences between the Sham and G-CSF groups or the CME and AG490 groups , with no significant differences between the former two groups or latter two groups  Fig. e. Notabl01) Fig. e, suggesups Fig. f. Similaups Fig. g, h. TakP < 0.01), but significantly smaller in the G-CSF group than in the CME or AG490 groups , but interstitial collagen content was significantly lower in the G-CSF group than in the CME or AG490 groups  and substantially higher in the G-CSF group than in the CME or AG490 groups (P < 0.01) , but these elevations were attenuated in the AG490 group  and electrical remodeling  that were accompanied by reduced levels of p-Cx43 and a redistribution of Cx43. Importantly, G-CSF partially or completely reversed these effects of CME, and the beneficial actions of G-CSF were attenuated by AG490, an inhibitor of JAK2-STAT3 signaling. Taken together, our findings suggest that G-CSF might act via the JAK2-STAT3 pathway to inhibit structural and electrical remodeling of the heart and enhanced susceptibility to ventricular arrhythmia following CME. This finding is another innovation of the present study.Most previous preclinical studies of CME used rat models induced by an autologous thrombus to simulate the rupture of a plaque in vivo –20, but Although MI and myocardial microinfarction may share some pathophysiological features , the mecG-CSF has received increasing attention as a cytokine that promotes repair after MI . G-CSF hThe mechanisms by which G-CSF exerts the above effects are unclear. Previous investigations reported that G-CSF causes bone marrow cells to migrate to infarcted regions and differentiate into cardiomyocytes and stimulates monocytes and neutrophils to promote neovascularization through paracrine mechanisms . On the The present study provides novel insight into CME, which affects 10–30% of patients and seriously affecting their prognosis . No drugThis study has some limitations. First, when generating the CME model, extraction of the microcatheter is challenging, and air embolism is more likely in rabbits than in larger animals because of narrow coronary arteries. Second, the patch-clamp technique was not used to directly measure the action potential duration of myocytes in the infarct site and peripheral zone, and the roles of ion channels were not investigated. Third, only AG490 was used , 37–39, In conclusion, our findings suggest that G-CSF acts via the JAK2-STAT3 pathway to inhibit the abnormal Cx43 expression, electrical remodeling, and enhanced susceptibility to ventricular arrhythmias that occur after CME.


The study was carried out between January 2013 and December 2018 at the Clinic of Urology . The protocol for all sEMG measurements of PFM activity consisted of following elements: “baseline”, “quick flicks”, “contractions”, “static hold”, and “rest tone”; we then compared these results between groups. To determine the optimal cutoff level for sEMG activation of the PFM to detect the occurrence of SUI, we performed receiver operating characteristic (ROC) curve analysis (with Youden’s index). Results: Significantly lower results were obtained for all PFM measurements in women with SUI. The optimum diagnostic cutoff for “baseline” was 3.7 μV (area under curve (AUC), 0.63), “quick flicks” was 9.15 μV , “contractions” was 11.33 μV , “static hold” was 9.94 μV , and “rest” was 3.89 μV . Conclusions: Measuring sEMG activity in the PFM may be a useful diagnostic tool to confirm the absence of SUI. We can expect that the sEMG activity of subjects with SUI will be lower than that of healthy people. In order to determine appropriate reference values for assessing sEMG activity data in the PFM, it is now necessary to conduct multicenter studies.Background: The use of surface electromyography (sEMG) measurements to evaluate the bioelectrical activity of the pelvic floor muscle (PFM) during therapeutic intervention is now well established. This study investigates the diagnostic possibilities of sEMG in women with stress urinary incontinence (SUI). The aim of this study was to carry out objective assessments of the bioelectrical activity of the PFM in women after menopause and determine the prognostic value of sEMG for assessing the PFM in patients with SUI. Methods: This was a prospective, observational study that evaluated the bioelectrical activity of the PFM in postmenopausal women with or without SUI (SUI group,  Previous literature highlights the validity of using surface electromyography (sEMG) measurements when assessing pelvic floor muscle (PFM) bioelectrical activity with regard to therapeutic progress ,3,4,5,6.This was a prospective, observational study that evaluated the sEMG activity of the PFM in postmenopausal women with or without SUI. The study protocol was approved by the Bioethics Committee of Wroclaw Medical University (KB-611/2012), and the study was conducted in accordance with the ethical principles of the Declaration of Helsinki. All patients provided written and informed consent.This study included women in their postmenopausal period with or without SUI and was carried out between January 2013 and December 2018 at the Department of Urology, University Hospital in Wroclaw (Poland). We recruited female patients from the department and volunteers who responded to a notice published in the local media. All recruited women were subjected to an evaluation that assessed specific inclusion and exclusion criteria. The exclusion criteria were as follows: aged over 75 years; no history of menopause; gynecological surgeries; surgeries in the abdomen/pelvis over the previous 10 years; injuries of the lower extremities, pelvis, or spine that were evident on the day of examination; prolapse of the reproductive organs; and third-degree incontinence ,12. PatiStudy participants were divided into two groups depending on the occurrence of urinary incontinence symptoms (SUI group vs. non-SUI group). The occurrence, or absence, of SUI symptoms was evaluated by an experienced urologist. In the urological examination, the urodynamic testing was included in order to determine the occurrence of urinary incontinence, its type, and its degree, and an additional International Consultation on Incontinence Questionnaire—Short Form (ICIQ-SF) was also used. The basis for qualifying for the SUI group was the involuntary loss of urine at least once a week for the preceding 3 months.The participants visited the clinic once at the beginning of the study, at which point a detailed medical history was collected. Prior to commencing the trial involving the use of sEMG to assess PFM performance, we assessed the correct action of the PFM during a contraction. Measurements were made in a supine position with bent lower limbs .Bioelectrical activity of PFM was assessed by surface electromyography and a compatible endovaginal electrode. The protocol of all measurements of PFM activity consisted of the assessment of the following elements: “baseline” , “quick flicks” , “contractions” , “static hold” (in which the participant attempted to hold the PFM contraction for 60 s), and “rest tone” .2/band (stainless steel). During measurements, the surfaces were directed to the right and left sides to record PFM activity with minimal crosstalk during tasks. This probe was inserted up to the handle at the introitus of the vagina. The monopolar reference electrode was placed on the anterior superior iliac spine.sEMG measurements were acquired using a MyoSystem 1400L eight-channel electromyographic apparatus . A Life-Care Vaginal Probe PR-02  vaginal probe was used to record the sEMG signal from the PFM. The pear-shaped probe has a total length of 7.6 cm and a maximal circumference of 2.8 cm. The length of the recording plate is 4.5 cm, and the active surface area is 7.68 cmThe electromyographic signal (EMG) signal was subjected to standard post hoc processing. First, data were rectified and smoothed using the root mean square (RMS) algorithm and then subjected to a filtering method to reduce the phase shift. A narrow band filter was used in the range from 50 to 1000 Hz (finite impulse response filter). The bioelectric activity of the muscles tested was expressed in microvolts.We paid particular attention to the following pitfalls related to sEMG measurements: (1) attention was paid to the exact, repeatable position during measurements; (2) we imposed an absolute ban on patient movement during the examination period; (3) the probe placement was carefully checked and performed in the same way for each participant; (4) the same type of probe was used for each examination; (5) in order to exclude additional pressure on the pelvic floor structures, the examination was performed with an empty bladder, and we also ensured that the patient had defecated and was not bloated on the day of examination. In addition, the patient was taught not to activate the abdominal press during PFM activity.t-test. We determined the following parameters: SUI group mean 9.4 μV; non-SUI group mean 14.2 μV; standard deviation 7.7 μV; null difference 0 μV. The alpha level was set at 0.05, and the power of the test was set to 0.9. On the basis of these parameters, the estimated sample size was 56 women for each group. In addition, we assumed that 10% of patients would withdraw from the study. The final sample size was defined as 62 participants per group.The estimated sample size was calculated in accordance with a pilot study . The mear ≥ 0.90). The distributions of baseline characteristics in the SUI group and the non-SUI group were compared with the chi-squared test for proportions and either the t-test or the Mann–Whitney U-test for continuous variables. Univariable and multivariable logistic regression were used to evaluate the influence of individual predictor variables, as well as their combined effect in predicting the absence of SUI. Variables in the univariate analysis that indicated a relationship to outcome (p ≤ 0.30) were selected for the multivariate logistic regression model (variables without multicollinearity). In cases where several variables had a correlation coefficient >0.6, a maximum of two of such variables were selected for the model . Statistically significant predictors were subjected to a backward, stepwise logistic regression analysis to determine which combination of predictors best explained the response. To determine the optimal cutoff level for sEMG activation of the PFM to detect the occurrence of SUI, we performed receiver operating characteristic (ROC) curve analysis (with Youden’s index). Based on the Youden index, the variable sEMG activities in the PFM were categorized (low vs. high) and included in a new logistic regression model. For all comparisons, we defined α as 0.05.Statistical analysis was conducted using Statistica 13 . In measurements where the study participant performed the test more than three times, the reliability and repeatability of these measurements for sEMG activity was assessed using an intraclass correlation coefficient (ICC). In each case, In total, 222 women were recruited and screened for this study; 80 women were excluded from the study for various reasons . Finallyp < 0.001). Significantly lower results were also noted in the assessment of sEMG activity during the “contraction” test , during the “static hold” test , at “baseline” , and at “rest” in women with SUI  and BMI  .The relationships between the absence of SUI and the sEMG activity of the PFM (continuous variables) and other variables , type of work, and the number of births) were determined by logistic regression analysis . The regROC curves were constructed separately for all sEMG tests  in ordern = 30) aged from 58 to 78 years  to validate the main results. On the basis of the results obtained, SUI was found in 17 participants. AUC results were compared between the study and validation groups. No statistically significant differences were found between the results. In the validation group the results were as follows: “baseline”—AUC, 0.62; 95% CI, 0.52–0.72; “quick flicks”—AUC, 0.84; 95% CI, 0.77–0.91; “contractions”—AUC, 0.79; 95% CI, 0.71–0.87; “static hold”—AUC, 0.85; 95% CI, 0.78–0.90; and “rest”—AUC, 0.63; 95% CI, 0.53–0.73.Additionally, a study was conducted on a prospectively assessed group of women , taking into account binary variables for sEMG .p < 0.001). In addition, the model also featured “quick flicks”    .We observed statistically significantly lower sEMG activities in the PFM of women with SUI. We also observed a higher odds ratio for the absence of SUI symptoms in women who obtained results above the cutoff point .Differences in the sEMG activities of the PFM between the SUI group and the non-SUI group represent important information in the context of SUI diagnostics. For example, Koenig et al.  evaluateIt should be noted that the results in these earlier articles show higher levels of activity in the PFM than observed in the current study; however, the mean age of the participants in the earlier studies was lower than in the present study. A large number of reports emphasize the fact that PFM activity decreases with age ,9,15,16.The existing literature shows that sEMG activities in the PFM differ markedly ,22,23,24The present study was limited by the fact that it was carried out in a single center. Applying the same research protocol in other centers would allow us to include a larger number of participants. It would also give us the possibility of including a wider range of population groups and the ability to compare results across centers. These factors would allow us to improve the generalizability of this study. Our results did not incorporate normalized sEMG test values. This is because normalization for the PFM assessment can be difficult and inappropriate and because we wanted to compare our data with data derived from other studies, which also presented non-normalized values.Our data suggest that measuring sEMG activity in the PFM may be a useful diagnostic tool to confirm the absence of SUI. We can expect that the sEMG activity of subjects with SUI will be lower than that of healthy people. In order to determine appropriate reference values for assessing sEMG activity data in the PFM, it is now necessary to conduct multicenter studies on a larger population, taking into account factors such as uniform research methodology, age, and the patient’s health status.


Promoting harm reduction programs as utilitarian in the public arena disregards their deeper prioritarian impulses. ‘Harm reduction’ is a misnomer, and the name should be abandoned sooner rather than later.‘Harm reduction’ programs are usually justified on the utilitarian grounds that they aim to reduce the net harms of a behavior. In this paper, I contend that (1) the  Harm reduction programs are not utilitarian, they are rarely implemented in a spirit of utilitarianism, they should not be promoted or defended as such, and the extension of ‘harm reduction’ into new arenas under the banner of utilitarianism is misguided. For most of the programs that bear the name, ‘harm reduction’ is a potentially damaging misnomer.Given the foregrounding of the words ‘harm’ (which implies an explicit assessment of harms and an implicit assessment of benefits) and ‘reduction’ (which implies a demonstrated decrease in harms), it is unsurprising that ‘harm reduction’ is generally considered a utilitarian approach to public health. Unsurprising, but wrong. In this paper, I contend that (1) the The key is to distinguish two responses to an unhealthy activity or risky health behavior. One response is to try to get people to stop behaving in that way, i.e., to try to get them to give up the activity in question. Policies with this intention are usually referred to as abstinence policies…The other response is to accept that, for whatever reason, it is unrealistic to expect people to give up the activity in question so it is better to allow it and try to reduce its harmful effects. Policies based on this response are known as ‘harm reduction’ .Precise definitions of ‘harm reduction’ are hard to come by, and the term has been subject to robust debate. Programs bearing the name encompass a variety of actors, practices, and goals , 11, andThere is some debate over whether harm reduction can ultimately be compatible with abstinence. Some claim that harm reduction should be sharply distinguished from abstinence programs ; others implicit in the reasoning of harm reduction advocates is a Utilitarian argument, which holds that the key guideline in ethics is that if negative consequences can be avoided they should be avoided. Abstinence advocates, however, generally seem to employ a Deontological ethic, which maintains that the moral worth of one’s actions has nothing to do with the consequences of those actions but, rather, is determined by the intention of the actors .Closely affiliated with this practical distinction between prohibition and acceptance is a conceptual one. As Christie et al. note, harm reduction is generally characterized as a consequentialist or utilitarian response to prohibitionist approaches that identify particular behaviors as wrong and seek to eliminate them:Similarly, Wodak defines harm reduction as “a consequentialist approach to drug policy, emphasizing the importance of outcomes rather than the policy intent,” and argues that the debate between its advocates and its critics “is essentially a conflict between ‘consequentialists’, more concerned to evaluate interventions by considering their impact, while the ‘non-consequentialist’ critics of harm reduction prefer to evaluate interventions by considering their moral worth” . Zajdow The centrality of consequences to the definition of harm reduction is often mobilized as an indication of its pragmatism and value-neutrality, and a contrast to moralizing abstinence-based approaches , 15, 17.An additional indication of the importance of utilitarianism within contemporary definitions of harm reduction can be found in the accounts of harm reduction’s critics. Supporters of abstinence-based programs openly reject consequentialist logic, in favor of deontologically-flavored arguments that drug use and other behaviors are categorically wrong . More recited as a core philosophical principle, when one looks more closely at the history and implementation of harm reduction, it is much harder to find evidence of utilitarianism as a moral imperative or guide to practice.While some form of utilitarian sensibility is almost universally The history of harm reduction is complex and multifaceted, and is characterized by “a historic tension between those who see harm reduction primarily as a medical means of promoting health and mitigating the harm to individual, and a more activist group who see it as a platform for broader and more structural social change” . While sanything that could potentially cause harms, and whose harms could be reduced, can be or already is subject to a ‘harm reduction’ program. Speed limits reduce the harms of driving. Seat belts and motorcycle helmets reduce the harms of traffic accidents. HPV vaccination programs reduce the harms of sexual behavior. Food regulations reduce the harms of eating and drinking. Social security reduces the harms of growing old. Maintaining a strong military reduces the harms of living in a world with antagonistic neighbors. Many possible policies could also be thought of in terms of harm reduction: Putting statins in the water could reduce the harms of poor diets, sedentary lifestyles, and possibly air pollution. Dispensing anti-depressants and mood stabilizers to employees could reduce the harms of stressful occupations. Conducting a pre-emptive strike on military bases within a hostile neighbor’s territory could reduce the harms of future disputes. One could plausibly make the case that any health or social policy that reduces the negative impact of a human behavior could be considered a ‘harm reduction’ program. But in practice this is not the case, and few if any of the interventions listed above are explicitly classified as such.On a conceptual level, it is difficult to see how utilitarianism and concern for consequences could possibly be a point that distinguishes ‘harm reduction’ from almost any other health or social policy . In theobehaviors that are illegal, socially unacceptable, or widely stigmatized; or harms to individuals or social groups who are stigmatized, marginalized, or generally worse-off. This fact—that the objects of harm reduction programs are chosen according to value-laden criteria, and not on the basis of dispassionate calculations of overall benefits and costs—is the first clue that the moral imperative behind harm reduction programs is anything but utilitarian. Programs do not provide clean needles and supervised injection sites (SIS) to injectable drug users, or publicly-funded ‘wet houses’ to alcoholics, because they have conducted a cost–benefit analysis and determined that these interventions would be an efficient use of resources, and we can certainly think of more efficient public health uses for these resources. If utilitarianism is the impetus behind these programs, it is a form of utilitarianism that is so heavily constrained or poorly conceived as to barely deserve the name.Most interventions reduce harm. What characterizes actual ‘harm reduction’ programs is that they reduce particular sorts of harm: either harms that result from The fact that harm reduction programs explicitly benefit the worst off is the clearest clue that their ultimate impetus is not utilitarian. They don’t simply ‘reduce harms’ on a population basis, they address the suffering of specific kinds of people. Indeed, if harm reduction programs were subject to a rigorous cost–benefit assessment—say, by comparing them to other interventions that would cost the same amount of money—they would likely fare poorly. This is not a bug of harm reduction programs, it is a feature, because what these interventions do that distinguishes them from other interventions is to address the suffering of the worst-off. Harm reduction is driven by compassion and a de facto prioritarian sense of justice, not a utilitarian calculus.One might reasonably contend that, even if the initial spirit of harm reduction was not utilitarian, its implementation surely is. Regardless of whether particular behaviors or groups are targeted, in practice these programs intend to, and actually do, reduce harms without making any sort of judgment about the morality of particular behaviors or individuals, which surely qualifies as the sort of value-neutral utilitarianism central to many of the definitions presented in the first section.This is wrong on two counts. First, harm reduction programs are selective about the exact harms that they reduce, and this selectivity poses problems for a utilitarian defense. SIS programs reduce overdose-related mortality, and transmission of communicable diseases through needle-sharing. They do not, however, reduce many of the other harms associated with illicit drug use, including the negative aspects of addiction itself; indeed, a successful abstinence program would arguably do a better job at harm reduction than any extant harm reduction program. If we are to be truly utilitarian about our assessment of these programs, it is worth asking whether, on balance, the harm reduced by preventing an overdose death actually outweighs the harms of continuing to use illegal drugs, living with an addiction, and causing continued suffering to those who are addicted and their families and caregivers.all the harms associated with particular behaviors [benefits of, for example, drug use are seldom accounted for [Second, harm reduction proponents routinely fail to fully account for ehaviors , 20. Harnted for , 8.For example, SIS aim  to reduce mortality associated with illicit drug overdoses. As it turns out, many of the feared costs of the programs—increased drug use, concentration of discarded needles, and so forth—have not materialized. On balance, then, SIS provide a net reduction of harms. But this conclusion is reached only by eliminating a number of other harms from consideration. The ‘mixed message’ criticism is often brushed aside, but one can certainly argue that it is harmful for a state to prohibit an activity, selectively enforce that prohibition, and at the same time devote resources to ensuring that individuals that engage in that activity can continue to do so. Most would agree that, if the state decided to employ a special emergency response team to ensure that drunk drivers who got in accidents received prompt medical attention, it would constitute a harmful mixed message. This harm may be difficult or impossible to identify; it may be so diffuse that its impact on any single individual is negligible; it may only apply to those who support the prohibition in the first place; it may be entirely debatable. But it is a harm. The diminution in property values or perceived quality of life of residents or businesses nearby SIS may be a harm that is merely financial or psychological, and impact very few, but these are still harms. Finally, by keeping users of a dangerous drug alive, the state implicitly supports producers and distributors of that drug—that is, it supports and incentivizes violent and antisocial criminals in their continued distribution of a harmful and addictive substance. This may well be due primarily to the fact that these drugs are criminalized, and perhaps decriminalization would reduce the harms they cause; but it is nevertheless clear that the drug and its distributors cause harms, and supporting them by keeping their customers alive is harmful. These are all harms, and if the philosophy of harm reduction is truly utilitarian then they should be counted. But they are not. Why?There are at least four grounds on which these and other harms may not be counted: theoretical impossibility, practical difficulty, extreme discounting, or irrelevance. We may dismiss the complaint of a parent uncomfortable with explaining to their child that the state prohibits drug use while simultaneously enabling it by claiming that it is not actually a consequence and thus can’t count as a ‘harm;’ by claiming that it is difficult or impossible to adequately quantifiable; by counting it but discounting it to such an extent as to render it irrelevant to any utilitarian calculus; or simply by ignoring it, because we do not see it as a problem.impossible to quantify. If it is the case that some critics of harm reduction are unconcerned with consequences, then perhaps the ‘harm’ of these programs results from the fact that they support behaviors that are intrinsically wrong, and cannot be considered in an instrumental manner within a consequentialist framework. This line of thinking is circular: we would rather not count objections to harm reduction as harms, so they must be deontological; and if they are deontological, we do not need to count these objections in a cost/benefit analysis. Moreover, many if not all of the criticisms of harm reduction programs could be expressed in terms of consequences. To use Campbell Brown’s apt term, we could ‘consequentialize’ objections to harm reduction, ‘count’ the costs to individuals or populations of (for example) tacitly condoning illegal behavior, and trade these costs off against the measurable benefits [First, they may also be benefits . This mapractically ‘uncountable.’ Harms that are primarily psychological or political are difficult to quantify, or at least far more difficult than the harms  that frequently are counted. It may be difficult to predict future possible harms like reduction of property values, and to isolate the contribution of an SIS to that reduction. This is not an unusual dilemma for utilitarians, and is closely related to the familiar aggregation problem—in this case, the difficulty of trading off clear and easily quantifiable harms for a small number of vulnerable individuals against diffuse and difficult-to-quantify harms to a larger and less vulnerable population. However, the practical difficulty of counting some harms does not disqualify them as harms, and a utilitarianism that simply ignores them because they are difficult to count is disingenuous.A second, more likely reason for the rigging of the calculus of harm reduction may be that many harms are can’t count some harms because it is impossible or prohibitively difficult, it is simply that they don’t count them.What I believe to be the most likely possibility is that many harms remain uncounted simply because harm reduction proponents choose not to count them. It is not that they do account for all possible harms, but simply weight many of them as zero or so infinitesimally small that they do not meaningfully impact a cost/benefit analysis of these programs. It is true that many objections to harm reduction programs are accounted for, and that these feared harms are often overestimated. Supervised injection sites do not increase discarded needles, drug use or initiation; promotion of contraceptive use does not increase sexual activity among students; and so forth. But the fact that the easily quantifiable benefits clearly outweigh the easily quantifiable harms is a happy accident, and does not imply that the unquantified harms will also have been overestimated.Failure to account for certain harms may occur within or outside of a utilitarian framework. The third possibility is that harm reduction proponents really are stringently utilitarian, and The fourth and most plausible explanation is that harm reduction proponents don’t count the harms proposed by unpalatable criticisms because they dismiss them entirely. For example, many harm reduction proponents  reject the ‘mixed messages’ argument because they believe that it relies on an illegitimate characterization of certain behaviors as ‘bad’, and thus unfairly stigmatizes behaviors or people as ‘bad.’ If drugs shouldn’t be illegal in the first place, how can it be harmful for the state to tacitly condone their use and provide clean needles and readily available naloxone? If teenagers having sex isn’t wrong in the first place, how can it be harmful for a school to acknowledge that teenagers might have sex and provide contraceptives to reduce unwanted pregnancies and STI transmission? Clearly, some assessment of right and wrong that takes no account of consequences guides these decisions about what counts as a ‘harm.’These examples illustrate a more general criticism that utilitarianism cannot stand on its own as a moral theory or guide to action, unless it is united with a theory of the good . Harm reContending that a philosophical framework is difficult or impossible to practically implement does not disqualify it as a useful tool to think with, even if it may consign it only to the realm of ideal theory. Utilitarianism is a useful means of encouraging us to dispassionately assess benefits and harms to the greatest extent possible, to acknowledge and confront the potential tradeoffs inherent in any policy or intervention, and to consider the competing claims to benefits and harms of different parties who are impacted—however tenuously—by these policies and interventions. The utilitarian justification of harm reduction is useful insofar as it helps us to confront these questions.raison d’être. Utilitarianism, which implies a dispassionate calculation of benefits and harms across an entire population, does not represent the ideal behind harm reduction programs, nor does it capture the essential quality that separates them from other public programs or policies that also reduce harms: an explicit selection of harms on the basis of moral commitments regarding who is harmed.Emphasizing the utilitarian aspects of harm reduction may also be an effective pragmatic strategy for justifying otherwise politically unpopular programs. Yet these contingent justifications are tactical, not essential, and we should not misread the public relations arm of a social movement as its Above, I demonstrated that harm reduction programs are not actually utilitarian in theory or in practice. Below, I will argue that this is fine, because the last thing in the world that we should do is justify the programs that currently bear the name “harm reduction” on the basis of utility or consequences. Utilitarianism is neither sufficient nor necessary for justifying harm reduction, and is ultimately detrimental.sufficient moral framework to justify harm reduction, because its practical implementation requires a prior choice of which harms and benefits count, and how they must be weighted. As noted above, this is a constituent feature of utilitarianism, which always requires some theory of the good to identify the appropriate harms and benefits. Yet utilitarian defenses of harm reduction programs tend to downplay or ignore these prior choices altogether, while emphasizing the dispassionate calculation. This gets it precisely backward. The substantive reasons for choosing which harms and benefits to count, among which people, and why, should take pride of place in justifications of harm reduction programs.Utilitarianism is not a unnecessary as a moral framework to justify harm reduction. Harm reduction programs are not, ultimately, about ‘reduction’ of ‘harms.’ They are about rejecting the punitive Manichaeanism that stigmatizes and admonishes those who engage in ‘bad’ behaviors. They are about rejecting the idea of moral desert, and showing compassion and respect for individuals, regardless of what kind of behavior they engage in. They are about making the lives of the worst-off slightly less terrible. We may argue over what philosophical framework best captures these impulses, but we can agree on one thing: utilitarianism isn’t it.1Utilitarianism is also It is worth asking why interventions rooted in justice and compassion for the worst off have been given the coldly utilitarian name of ‘harm reduction.’ On one hand, it is a pragmatic response to complex problems whose solutions may not materialize any time soon. Rather than waiting for an end to the War on Drugs, or a sea change in attitudes towards sex, we adopt practical programs to make peoples’ lives a little less bad. On the other hand, it might be a recognition of the meanness of our political moment: in a context of social division and skyrocketing inequality, rejection of Manichaeanism and compassion for the worst-off—particularly stigmatized groups like drug users and sex workers—is so unpopular that we are afraid to invoke it even in the context of public health. We feel that we must frame compassion as calculation, interpersonal warmth as impersonal coldness.2If this is the case, if utilitarianism is simply a pragmatic public relations ploy, some might (aptly) wonder, what’s the harm with ‘harm reduction’? Indeed, shouldn’t we capitalize on the success of extant harm reduction programs by extending their logic to other arenas? So long as these programs help without causing meaningful harms, does it matter what we call them? I think that it does.all the harms, and found them to outweigh the benefits? What if, for example, provision of free naloxone for drug overdoses increases drug use?First, if you live by the utilitarian sword you may also die by it. It so happens that the tradeoffs that have so far been identified have come out in favor of harm reduction programs; but what if they don’t? What if we counted Second, I think that framing compassionate programs as utilitarian ones ultimately contributes to the continued stigmatization of behaviors and dehumanization of the people that they seek to help. Defending compassionate attempts to address the suffering of the worst-off on the utilitarian grounds that they reduce overall harms, rather than the grounds that they are human beings whose suffering must be addressed, diminishes their moral force. We do not justify extraordinary interventions to help children with leukemia, or to rescue a teenage soccer team trapped in a cave, on the grounds that they reduce harm or improve overall utility—for surely they do not. Why then should we justify helping sex workers or injectable drug users on these grounds, unless we already tacitly accept that they do not ‘deserve’ our help in the same way that child cancer patients or teenage soccer players do?Finally, I think that framing compassionate programs in utilitarian terms unnecessarily, and wrongly, cedes the ‘moral high ground’ to critics. ‘Harm reduction’ is often pitched as the value-neutral, less-controversial label for programs that might be unpopular because they are associated with ‘bad’ behaviors or ‘bad’ people. This might be politically pragmatic, but it is ultimately harmful because it implicitly accepts the designation of these behaviors and people as ‘bad’ rather than rejecting these labels, challenging the moralizing, and offering the positive argument that human beings deserve respect and compassion. Claiming an instrumental ethos for harm reduction invites opponents to claim an intrinsic motivation which in the popular arena reads as “proponents of harm reduction just care about numbers, opponents care about what’s right.” This claim is tendentious, but so long as defenders of compassionate programs embrace instrumentalism it will persist.The rightness of ‘harm reduction’ programs—and what distinguishes them from other public health interventions—has nothing to do with harm and nothing to do with reduction. Their rightness inheres in rejecting Manichaeanism and moral desert, and respecting and giving priority to the lives of the worst off. It is not and should not be subject to a consequential calculus, and we should not pretend that it is or should be. We should not cede the territory of rightness to the critics; we should instead forcefully proclaim the conviction that helping those who suffer is intrinsically right. ‘Harm reduction’ is a misnomer, and the name should be abandoned sooner rather than later.