Tumour infiltrating lymphocytes (TILs) are a prognostic parameter in triple-negative and human epidermal growth factor receptor 2 (HER2)-positive breast cancer (BC). However, their role in luminal (oestrogen receptor positive and HER2 negative (ER + /HER2-)) BC remains unclear. In this study, we used artificial intelligence (AI) to assess the prognostic significance of TILs in a large well-characterised cohort of luminal BC.Supervised deep learning model analysis of Haematoxylin and Eosin (H&E)-stained whole slide images (WSI) was applied to a cohort of 2231 luminal early-stage BC patients with long-term follow-up. Stromal TILs (sTILs) and intratumoural TILs (tTILs) were quantified and their spatial distribution within tumour tissue, as well as the proportion of stroma involved by sTILs were assessed. The association of TILs with clinicopathological parameters and patient outcome was determined.A strong positive linear correlation was observed between sTILs and tTILs. High sTILs and tTILs counts, as well as their proximity to stromal and tumour cells (co-occurrence) were associated with poor clinical outcomes and unfavourable clinicopathological parameters including high tumour grade, lymph node metastasis, large tumour size, and young age. AI-based assessment of the proportion of stroma composed of sTILs  was not predictive of patient outcome. tTILs was an independent predictor of worse patient outcome in multivariate Cox Regression analysis.AI-based detection of TILs counts, and their spatial distribution provides prognostic value in luminal early-stage BC patients. The utilisation of AI algorithms could provide a comprehensive assessment of TILs as a morphological variable in WSIs beyond eyeballing assessment. Breast cancer (BC) is a heterogeneous disease with different molecular subtypes and variable clinical behaviours . DespiteThe role of tumour infiltrating lymphocytes (TILs) in refining BC prognosis and possible targeted immunotherapy has been widely studied –8. It haTo date, it is recommended that visual assessment of TILs in routine clinical practice depend solely on stromal TILs (sTILs), defined as the percentage of mononuclear cells (lymphocytes and plasma cells) quantifiable in the stromal area . AlthougThe widespread use of digital pathology and the increasing applications of artificial intelligence (AI) on whole slide images (WSIs) –20 have In this study, we hypothesised that using AI-based algorithms would provide improved assessment of TILs in early-stage luminal BC patients and identify additional TILs features that have prognostic implications.This study was conducted on two cohorts including:n = 1572) and test (n = 659) sets using stratified random sampling to ensure equal distribution of events in each set. Initially, the discovery set underwent an internal 3-fold cross-validation, where the set was stratified into three different splits which were used for training the model. Then the optimised module resulting from this cross-validation process was applied to the test set. To note, the test set remained untouched throughout the AI model upstream steps to ensure a reliable validation. However, for the simplification of the results, the correlation between TILs against the clinicopathological, and outcome data was carried out on the whole discovery set and validated on the test set.A) Nottingham cohort: a cohort of 2231 endocrine-treated ER + /HER2- BC patients presented to Nottingham City Hospital, Nottingham, United Kingdom. Anonymised clinicopathological data including patient’s age, menopausal status, tumour size, histological subtype, tumour grade, Nottingham prognostic index (NPI), lymph node (LN) status, lymphovascular invasion (LVI), PR and Ki67 expression scores, as well as treatment information were collected. All patients included were treated with adjuvant endocrine therapy only based on local treatment protocol; ER + BC patients were offered chemotherapy only if the NPI score is in the poor prognostic group and the patient tolerates chemotherapy  interval and different augmentation methods  were used during training with values of 0.5. To restrict stromal region only to TAS instead of overall stroma, the following steps were performed: i) stromal regions segmented by CNNReg were combined with tumour regions segmented by CNNDCIS; ii) to capture the TAS, the tumour regions were dilated with a disc of radius 8 pixels. This was followed by filling the holes with a disc of radius 32 pixels; iii) any stroma captured in the final dilated tumours was considered as TAS for further feature calculation. Immune nuclei were then counted separately in the tumour regions as well as in the TAS. Other features included co-occurrences of different nuclei, immune heterogeneity, and contrast.Quantification of immune cells within tumour and TAS requires region segmentation. For the purpose of sematic segmentation of the regions, a CNN model known as the U-Net  was adapNuc based on HoVer-Net [rd epoch onwards), patch size 256 × 256 pixels and batch size 8. During inference on a WSI, a nuclei mask was generated by mapping the type/class of each detected nucleus at its centroid in a five-times down sampled WSI. Combined with segmented tumour and TAS regions, the counts of immune nuclei were used to get the tTILs and sTILs counts  was scored by a well-trained pathologist as a continuous parameter following the proposed scoring method by TILs Working Group [A subset of cases  was considered significant in all the statistical tests.Analysis was performed using Statistical Package SPSS v28 for Windows . Assessed features were used as continuous variables and association with clinicopathological parameters was evaluated using Mann-Whitney and Kruskal-Wallis tests. Each feature was dichotomised, into low and high for outcome analysis based on X-tile software version 3.6.1   using BCAnalytical performance of different ML modules used in TILs quantification and distribution are shown in Supplementary Table AI-based sTILs percentage had a range from 1% to 76%, which was close to results obtained by visual assessment (ranged from 0 to 80%). The concordance between the visual and AI-based sTILs percentage, showed good agreement (ICC = 0.7).p < 0.001) and this correlation was observed in the discovery and test sets. Similarly, the sTILs count and AI-based sTILs percentage showed significantly positive correlation  in both sets. In the external validation cohort, the correlation between sTILs and tTILs count and between sTILs count and AI-based sTILs percentage was positive . The median count of TILs and TILs co-occurrence scores in WSIs, was calculated in the study cohorts  than tumours with low sTILs  .Patients with BC showing high sTILs had significantly shorter survival   .Multivariate Cox Regression analysis of all significant features, adjusted for tumour grade, LN metastasis, Ki67 index and tumour size, revealed that tTILs count is an independent predictor of outcome. Every 1000 unit increase in tTILs count is associated with 20% increase in BC- related death risk  in predicting patient outcomes, and the possible influence of chemotherapy and hormonal treatment on TME has been demonstrated . LuminalIt has been proposed that TILs abundance is sufficient to be assessed in TNBC to guide a prognostic benefit, while in luminal tumours, biological stratification of TILs should be more informative , 47. HowThe use of the readily available H&E WSIs in such automated analyses will enable more cost effective use of tissue and clinical resources . AutomatAlthough most previous studies did not focus on tTILs due to their lower density compared to sTILs , interesAutomated TILs assessment could provide data about the distribution of TILs in relation to stromal and tumour cells. Here the histological ecology of TILs, or how TILs interact with their neighbouring tumour and stromal cells, was investigated in terms of their spatial distribution . It has One of the features that can be generated by AI algorithms is the TILs count, which may be a more powerful prognostic indicator than the usual routine assessment method. In relation to patient outcome, TILs count was significantly associated with poorer survival, unlike the sTILs percentage scored both manually and AI-assisted, which lacked prognostic significance. This was consistent with previous studies that reported high TILs were negatively associated with recurrence-free survival and distant metastasis-free survival , 63. It TILs counts were significantly associated with unfavourable clinicopathological parameters in this study in line with the findings of former studies , 63. HigWe propose that one of the contributors to disagreement between studies was the use of different methodologies, whether total TILs or subclasses quantification through IHC detected subpopulations or using different cut-off values for defining low versus high TILs. In our analysis, the TILs count was used as continuous variables to avoid a biased cut-off point.Our cohort was endocrine treated with poor prognostic outcomes of tumours enriched with TILs. Dunbier and colleagues reported poor response to aromatase inhibitors in ER+ tumours rich TILs suggesting that TILs are involved in resistance to hormonal therapy . MoreoveWe recognise the study has some limitations. Firstly, this study was conducted on H&E-stained sections only which did not allow for the identification of different immune subpopulations and their distinct roles. In routine practice, H&E sections are the standards for TILs evaluation and its association with tumour behaviour and patient outcome, however, the identification of the makeup of the various subpopulations would help in understanding the underlying biology. Various immune cells, including CD8 + , CD4 + , FOXP3 T lymphocytes, B lymphocytes, plasma cells and macrophages (including M1 and M2 subtypes) would have different roles in tumour microenvironment and behaviour. The predominate immune cell type may exert significant control over the tumour behaviour. Secondly, although our model showed high accuracy in identifying and distinguishing TILs from other cell types, we must acknowledge the potential presence of false TILs identification. As with any automated or AI-based system, there is an inherent risk of misclassifications. Thirdly, although using large well characterised cohort, the small number of events in our study warrants careful interpretation of the results and further validation utilising external independent cohorts with long term follow up is recommended.In conclusion, automated assessment of both TILs counts, and spatial distribution provides independent prognostic value in early-stage luminal BCs. The utilisation of AI algorithms would add to the limited information pathologist can retrieve from visual assessment.Supplementary materials


DNA methylation (DNAm) plays a crucial role in a number of complex diseases. However, the reliability of DNAm levels measured using Illumina arrays varies across different probes. Previous research primarily assessed probe reliability by comparing duplicate samples between the 450k-450k or 450k-EPIC platforms, with limited investigations on Illumina EPIC arrays. We conducted a comprehensive assessment of the EPIC array probe reliability using 138 duplicated blood DNAm samples generated by the Alzheimer’s Disease Neuroimaging Initiative study. We introduced a novel statistical measure, the modified intraclass correlation, to better account for the disagreement in duplicate measurements. We observed higher reliability in probes with average methylation beta values of 0.2 to 0.8, and lower reliability in type I probes or those within the promoter and CpG island regions. Importantly, we found that probe reliability has significant implications in the analyses of Epigenome-wide Association Studies (EWAS). Higher reliability is associated with more consistent effect sizes in different studies, the identification of differentially methylated regions (DMRs) and methylation quantitative trait locus (mQTLs), and significant correlations with downstream gene expression. Moreover, blood DNAm measurements obtained from probes with higher reliability are more likely to show concordance with brain DNAm measurements. Our findings, which provide crucial reliable information for probes on the EPIC array, will serve as a valuable resource for future DNAm studies. Alteratcytosine –6. Methycytosine . Althougcytosine , 9.In recent years, several studies have examined the reliability  of DNAm levels from the same DNA samples measured twice using Illumina arrays, and found that probe reliability varies across different probes –15. HoweTo address these critical gaps and assess the reliability of blood DNAm levels measured using EPIC arrays, we conducted an analysis using 138 duplicated blood DNAm samples from the Alzheimer’s Disease Neuroimaging Initiative (ADNI) study . We compadni.loni.usc.edu.We analyzed a subset of whole-blood DNAm samples generated by the Alzheimer’s Disease Neuroimaging Initiative (ADNI) study , in whicDNA methylation was measured using the Illumina HumanMethylation EPIC BeadChip, which includes more than 850,000 CpGs. We preprocessed the DNAm data using the SeSAMe 2 pipeline described by Welsh et al. (2023), which was found to perform the best and produced the largest percentage of reliable CpG probes in a recent comparison of various preprocessing and normalization pipelines . Supplemp-value threshold of 0.01) in the RnBeads R package, which iteratively removes the probe or sample with the highest fraction of unreliable measurements one at a time [P-value with out-of-band array hybridization) algorithm in SeSAMe R package in more than 20% of samples. Finally, we performed a noob  background correction and nonlinear dye-bias correction [First, we removed CpGs that overlapped with single nucleotide polymorphism (SNP), non-CpG probes, cross-reactive probes , and prot a time . Next, wTo estimate the reliability of CpG probes, we computed intraclass correlations (ICCs) for each probe based on methylation beta values, which were measured in duplicates of blood samples collected from the same subject and at the same visit. The ICC is defined as i (2016) , ICC valthe ICC – half-width of 95% confidence Limits of Agreement (HoLA). The HoLA was calculated as As discussed below, probes with a large between-subject variance could yield high ICC values, even in the presence of conspicuous differences between duplicate measurements. To explicitly account for the amount of disagreement in duplicate measurements, we proposed to assess reliability using a modified ICC, which is defined as To compare the reliability of probes with different characteristics , we performed mixed-effects model analyses using the lmerTest R package . For eachttp://mqtldb.godmc.org.uk/downloads. To select significant blood mQTLs in GoDMC, we used the same criteria as in the original study [P-value smaller than 10−8 and a trans P-value smaller than 10−14 as significant. For DNAm-to-mRNA association analysis, we analyzed matched gene expression (Affymetrix Human Genome U 219 array) and DNA methylation (EPIC array) data from 263 independent subjects in the ADNI study. In this analysis, we examined probes located in promoter regions  and distal regions (> ± 2k bp of the TSS) separately. Specifically, for CpGs located in the promoter region, we computed Spearman correlations between CpG methylation and expression levels of the target genes. On the other hand, for CpGs in distal regions, we computed the Spearman correlations between CpG methylation and expression levels of ten genes upstream and downstream, following the approach used in previous integrative DNAm-to-gene expression analyses for probes in distal regions [P-value for each probe and considered P-values less than 1 × 10−5 to be statistically significant, as in several previous analyses of DNA methylation in blood samples [We searched mQTLs for CpG probes using the GoDMC database , which wal study , that is regions . Subsequ samples , 28. To  samples .The estimated ICCs for individual probes are available in Sugden et al. (2020) , Logue eP-value < 0.001, Supplementary Figure 1). A comparison of the 333,588 common probes in both studies showed that a larger number of probes achieved good reliability  and excellent reliability  in our EPIC-EPIC comparison study compared to the 450k-EPIC comparison in Sugden et al. (2020) study  (Supplementary Table 3). Overall, 214,951 probes (64.44%) had the same classification in both the studies.The ICCs for probes in our EPIC-EPIC comparison ranged from −0.362 to 0.999, with a mean of 0.381 and a median of 0.325 . The ICCIn contrast, the ICCs in the EPIC-EPIC comparison were only slightly higher than those in the 450k-450k comparison estimated by Bose et al. (2014), which ranged from 0 to 0.998, with a mean of 0.366 and a median of 0.296  . Given that a substantial proportion of type I probes are located in CpG-rich regions, such as CpG islands and promoter regions [We found that the distribution of ICC values in our EPIC-EPIC comparison showed patterns similar to those of previous comparisons –13, 15. n levels . The hig regions , 31, thith (0.18%) among the 640,960 probes. Therefore, some probes, such as cg14089881, can achieve high ICC values because of their relatively high between-subject variance, As noted previously , 32, a cHalf-width of the 95% confidence Limits of Agreement (HoLA). We define the modified ICC as the ICC subtracted by HoLA, which penalizes the ICC by the amount of disagreement in duplicate measurements for the probe. Note that the modified ICC prioritizes probes with a large ICC and small HoLA. On the other hand, probes with a large ICC and substantial disagreement in duplicate measurements will have a reduced modified ICC compared with the ICC. For example, the modified ICC for cg14089881 mentioned above was 0.5579, indicating only fair reliability for this probe after correction for disagreement based on HoLA.Therefore, we propose a modification to ICC by incorporating the In the following sections, we will assess the impact of probe reliability, as measured by the modified ICC, in relation to different types of analyses in epigenome-wide association studies (EWAS).P-value < 2.2 × 10−16). Specifically, CpG probes influenced by mQTLs had a median modified ICC of 0.546 compared to the other CpGs with a median modified ICC of 0.214.We hypothesized that probe reliability might affect the effectiveness of integrative analyses that correlate DNAm with other types of omics variants, such as mQTLs or downstream gene expression. Methylation Quantitative Trait Locus (mQTLs) refers to genetic variations that influence the patterns of DNAm. Min et al. (2021) performed a large mQTL study involving 32,851 subjects and found that approximately 45% of DNAm sites on the Illumina array were influenced by genetic variants . SupplemP-value = 3.24×10−5 and P-value = 1.41×10−9, respectively; Supplementary Figure 13). These findings highlight the importance of considering probe reliability in EWAS and its potential implications for understanding the relationship between DNAm, genetic variation, and gene expression.Similarly, as DNAm is a key epigenetic modification that influences gene activity by regulating gene expression, we next investigated the impact of probe reliability on DNAm-to-mRNA correlations in blood samples. In this analysis, we examined probes located in promoter regions  and distal regions (> ± 2k bp of the TSS) separately. For both groups of probes, we found that the modified ICCs were higher for those probes significantly associated with downstream gene expression levels compared to other probes  examined matched DNAm profiles of pre-mortem blood samples and post-mortem brain tissues in the London dataset and found that only a small proportion of CpGs showed significant brain-blood correlations in DNAm levels .To assess the impact of probe reliability on cross-tissue associations, we examined modified ICCs for the probes in relation to the correlation of DNAm measured in the brain prefrontal cortex and blood in the London dataset. Supplementary Figure 14 demonstrates that probes with higher brain-blood correlations in DNAm levels also had higher reliability. Specifically, for probes with high (> 0.75), medium (0.4–0.75), and low (< 0.4) brain-blood correlations, the median modified ICCs were 0.869, 0.786, and 0.215, respectively. These results are consistent with another recent study, which found an increase in reliability estimates in the 450k-EPIC comparison [A previous study by Sugden et al. (2020) analyzed the effect of tobacco smoking on DNAm across 22 studies and observed that the number of replications of individual probes across studies positively correlated with reliability . We prevP-value = 0.049). Specifically, the median modified ICC for probes within DMRs was 0.726, which was significantly higher than the median of 0.275 observed for the other probes. We observed similar patterns of association in sex-specific analysis of AD-associated DMRs .Biologically, DNAm levels are often correlated across the genome and occur as a regional phenomenon . DiffereThese findings highlight the importance of probe reliability in studying DNAm associations, as reliable probes help to minimize discrepancies between different datasets and lead to more consistent results. Therefore, probe reliability is crucial for detecting genuine DNAm associations in EWAS.In EWAS, one common approach for accounting for cell-type heterogeneity across different samples is to estimate the proportions of various cell types within each sample. These estimated cell type proportions were then included as covariates in the regression models. We assessed the reliability of these estimated cell type proportions and found that they showed good agreement between duplicate samples (Supplementary Table 5). Specifically, the modified ICC ranged from 0.693 to 0.930 across the different cell types. Notably, the proportions of NK  cells and B cells were observed to have the lowest and highest reliabilities, respectively. These results are consistent with a previous study that examined DNAm levels in newborn and 14-year-old samples, which reported a high correlation in estimated cell-type proportions between duplicate samples . Taken tIn this study, we comprehensively evaluated the reliability of probes on Illumina EPIC arrays and created a valuable resource for EWAS studies (Supplementary Table 6). We carefully selected DNAm samples from the ADNI dataset. To avoid batch effects and ensure accurate assessment of probe reliability, we included duplicate samples that were placed on different methylation plates within the ADNI dataset.Compared to previous studies that estimated probe reliability using duplicate measurements on either the 450k-EPIC or 450k-450k arrays, we found that the overall reliability estimates were higher in our study, in which both duplicates were measured using the EPIC arrays, consistent with results from previous studies , 17  in the SeSAMe R package. This step specifically identifies and removes probes with hybridization issues [It is important to consider additional factors that contribute to probe reliability. During our preprocessing step, we incorporated the pOOBAH algorithm  , we alsoOn the other hand, probes with large between-subject variance could yield high ICC values, even in the presence of sizable differences between duplicate measurements. To explicitly account for the disagreement in duplicate measurements, we proposed a modified ICC that incorporates a penalty based on the half-width of the 95% confidence limits of agreement. We found that the modified ICC maintained the characteristics of the original ICC, with similar patterns of association with the mean and standard deviations of beta values. It also had lower values in CpG islands and TSS200 regions, as well as for CpGs measured by type I probes. The classification of probes based on the modified ICC largely agreed with the classifications based on the original ICC, except for probes with substantial disagreement in duplicate measurements.We next studied the implications of probe reliability in the EWAS, and our findings highlight its significant impact on downstream analyses. Consistent with observations in previous studies that used ICCs to assess reliability , 15, ourSeveral limitations and areas for future study are in order. First, our analyses focused on methylation levels measured in whole blood; therefore, the results may be applicable only to EWAS conducted using blood samples. Additional studies that assess the reliability of DNAm measured by EPIC array probes in target tissues  and other accessible tissues  are needed. Second, due to the specific criteria we applied, including selecting independent subjects and ensuring that samples were placed on different methylation plates, we were able to include only a relatively small number of 69 pairs of duplicate samples in this study. Future studies with larger sample sizes are needed to confirm our findings. Finally, it is important to note that reliability estimates can be influenced by the choice of normalization procedure. In this study, we used the SeSAMe 2 pipeline, which was found to have the best performance and yielded the largest percentage of reliable CpG probes in a recent comparison of various preprocessing and normalization pipelines in a small study with 16 pairs of duplicate samples . Additioa priori [To reduce the burden of multiple comparisons in EWAS, some authors have proposed excluding probes with low reliability a priori , 40, whea priori . To thisparisons . The priIn summary, we comprehensively evaluated the reliability of probes on EPIC arrays. We observed that probes with substantial between-subject variance could still yield high ICC values even when there were notable differences between duplicate measurements. To explicitly account for the disagreement between duplicate measurements, we propose a new statistical measure, the modified ICC. Our findings revealed that probe reliability has significant implications for various downstream analyses of the EWAS. Importantly, we generated a valuable resource for DNAm research by identifying a set of high-quality probes on the EPIC array, which will contribute to optimizing the robustness and potential of EWAS.


Prostate cancer is the second most frequent malignancy in men worldwide and the fifth leading cause of death by cancer. Although most patients initially benefit from therapy, many of them will progress to metastatic castration-resistant prostate cancer, which still remains incurable. The significant mortality and morbidity rate associated with the progression of the disease results mainly from a lack of specific and sensitive prostate cancer screening systems, identification of the disease at mature stages, and failure of anticancer therapy. To overcome the limitations of conventional imaging and therapeutic strategies for prostate cancer, various types of nanoparticles have been designed and synthesized to selectively target prostate cancer cells without causing toxic side effects to healthy organs. The purpose of this review is to briefly discuss the selection criteria of suitable nanoparticles, ligands, radionuclides, and radiolabelling strategies for the development of nanoparticle-based radioconjugates for targeted imaging and therapy of prostate cancer and to evaluate progress in the field, focusing attention on their design, specificity, and potential for detection and/or therapy. Prostate cancer (PCa) is the second most frequent malignancy in men worldwide and the fifth leading cause of death by cancer, with a near estimate of 1.4 million new cases and 375,000 deaths worldwide . Accordi99mTc-labelled bisphosphonate bone scintigraphy (BS) Ga-PSMA-11 or [18F]F-DCFPyL is approved by the US Food and Drug Administration (FDA) and recommended in the Guidelines of the European Association of Urology as an imaging technique of choice for the majority of patients with biochemically recurrent PCa after curative treatment and for assessment of metastases Tc-PSMA agents have been explored for PCa imaging and radio-guided surgery. Although clinical studies have shown that these molecular probes possess high sensitivity and specificity in the initial diagnosis of PCa, monitoring of disease progression and response to therapeutic treatment, none of them exceeds the accuracy and sensitivity of the PSMA PET radiopharmaceuticals Lu-PSMA-617 (PluvictoTM) was approved by the FDA for the treatment of patients with PSMA-positive mCRPC who have previously failed other therapy options [177Lu]Lu-PSMA-617 [177Lu]Lu-PSMA-617 is not efficient for the treatment of micrometastatic disease and circulating tumour cells because β− particles emitted from 177Lu have a relatively long range in the soft tissue (1–10 mm), well beyond a diameter of micrometastases CuS) (64Cu]CuS radioconjugate and untargeted PEG-[64Cu]CuS) radioconjugate, and determined their specific binding and cellular uptake in vitro into aggressive prostate cancer cells PC-3-KD1. The ability of these radioconjugates to accumulate in orthotopic prostate tumours was further evaluated via PET imaging in nu/nu mice bearing PC-3-KD1 tumours. The in vitro results showed that bombesin-targeted showed a much faster cellular binding rate and far better uptake efficiency compared with untargeted NPs.In 2018, Cai et al.  presente4Cu]CuS) . The aut64Cu]CuS NPs had the potential to reduce a nonspecific uptake of NPs by the mononuclear phagocytic system.The PET/CT imaging revealed that in vivo uptake of untargeted NPs by tumour tissue was very low and ranged from 1.2 and 1.4% (ID/g) after 1 and 6 h, respectively. In contrast, the bombesin-targeted radioconjugate gradually accumulated in the tumour tissue, displaying enhanced tumour-to-surrounding tissue contrast and significantly higher tumour accumulation levels . Analysis of biodistribution confirmed the PET/CT data and showed that both bombesin-targeted and -untargeted NPs were highly uptaken by liver. Moreover, untargeted NPs displayed significantly higher uptake than bombesin-targeted NPs in the spleen, suggesting the active targeting capability of the Bom-PEG-PEG-(DFB)3, [89Zr]PEG-(DFB)3(ACUPA)1 and [89Zr]PEG-(DFB)1(ACUPA)3 were tested in PSMA-positive PC3-Pip and PSMA-negative PC3-Flu cells. In vivo PET imaging of the 89Zr-labelled nanocarriers and organ biodistribution studies were performed in the nu/nu athymic mice bearing subcutaneous dual xenografts of PC3-Pip and PC3-Flu tumours. In vitro results revealed that the use of multivalent PSMA binder [89Zr]PEG-(DFB)1(ACUPA)3 with three copies of ACUPA ligands increased PSMA binding affinity and cellular internalization compared with the conjugate containing the single ACUPA ligand.In 2022, Meher et al.  designedchelator . In vitr89Zr]PEG-(DFB)1(ACUPA)3 bearing three ACUPA ligands showed a remarkably higher PC3-Pip/blood ratio and background clearance.In vivo and ex vivo studies revealed that both conjugates demonstrated remarkably higher tumour retention and background clearance in PSMA-positive PC3-Pip tumours compared to nontargeted conjugates. Although targeting significantly improved tumour retention and tissue penetration of both nanocarriers in PSMA-positive PC3-Pip xenografts, the multivalent nanocarrier -AuNP-Lys3-bombesin conjugate in tumour, liver, and spleen. The results revealed a 1.5 times higher uptake by the tumour tissue and tumour-to-blood ratio for the targeted NP-based conjugate than for targeted NP-free [99mTc]-HYNIC-Lys3-bombesin conjugate.In vitro binding studies revealed that the NPs are able to specifically recognise the GRP receptors overexpressed in prostate cancer PC-3 cells. Cellular uptake of the targeted NPs ] in nude mice bearing PC-3 tumour showed ~six-fold higher tumour uptake after 24 h post-injection compared with non-targeted AuNP-DTDTPA-67Ga.Biodistribution studies of this radioconjugate -PSMA-PLGA-CUR accumulated specifically in the tumour tissue in a dose-dependent manner in the LNCaP C4-2 xenograft mice model. The whole-body and ex vivo imaging of multiple organs proved that the NP-based radioconjugate retained within the tumour tissue at a much higher level than the NP-free [131I]-PSMA targeting antibody. In contrast to the tumour, other organs exhibited minimal 131I radioactivity from the NP-based radioconjugate.The results indicated that these NPs efficiently inhibited the growth of PCa cells both in vitro and in vivo. The NP-based radioconjugate [111In by DOTA chelation. The hydrophobic antiangiogenic drug sorafenib was loaded onto these nanoparticles for chemotherapeutic applications  and radiolabelled with 64Cu [64Cu]DOTA-cys-DB-PEG-LNP, and NP-free doxorubicin radiolabelled with 64Cu [64Cu]DOTA-Dox  compared to the NP-based untargeted [64Cu]DOTA-Dox-PEG-LNP radioconjugate (10% ID/g) and NP-free untargeted [64Cu]DOTA-Dox radioconjugate (3% ID/g). Moreover, a four-fold increase in tumour uptake was observed for NP-based untargeted radioconjugate containing doxorubicin [64Cu]Dox-DOTA-PEG-LNP (10% ID/g) compared to NP-free untargeted doxorubicin [64Cu]-DOTA-Dox (3% ID/g).A pharmacokinetic study revealed that blood clearance of a covalent mixture of NP-based [3), comprising a small molecule Lys–urea–Glu-based PSMA inhibitor (PSMAL), PEG2000, and palmitate linker, and radiolabelled with 99mTc radionuclide or loaded with doxorubicin . The purpose of this study entitled, “Molecular Phenotyping and Image-Guided Surgical Treatment of Prostate Cancer Using Ultrasmall Silica Nanoparticles”, is to determine the usefulness of [225Ac and 223Ra). This prevents daughter radionuclides from escaping from the target site and reduces toxic side effects. In addition, nanoparticles allow overcoming difficulty in the stable attachment of some radionuclides  to the targeting ligands. Moreover, they allow encapsulation and delivery of drugs , sorafenib, and curcumin) into the tumour. The in vivo results confirmed the controlled release of the drug over a long period of time, increasing therapeutic index, accumulation into the tumour tissues, and inhibition of the tumour growth. Furthermore, the most important advantage of some NP-based radioconjugates is the possibility to deliver the drug and imaging agent(s) at the targeted site to diagnose and treat the PCa. Despite these many advantages of the NP-based radioconjugates, there are still some disadvantages/challenges hampering their translation to the clinic. One of the biggest problems in the use of NP-based radioconjugates is their entrapment in mononuclear phagocytic systems. Surface modification of NPs with polyethylene glycol (PEG) prevents their agglomeration and results in prolonged presence in circulation due to hindered recognition and phagocytosis, but this strategy is not efficient against accumulation in non-tumour tissues. The next important disadvantage of these radioconjugates is the lack of adequate knowledge about their long-term impact on cellular signalling pathways and biochemical processes of the human body. Another disadvantage is poor control of degradation, pharmacokinetics, and biodistribution in vivo. Therefore, for each promising NP-based radioconjugate evaluated in preclinical studies in vitro, the risk-to-benefit ratio should be determined in vivo before introducing them to the clinic.Recent developments in the field of NP-based radioconjugates for targeted imaging and therapy of PCa have revealed a wide diversity of design approaches and methods of preparation. The most important advantage of these radioconjugates is highly specific targeting . This pr


Inappropriate antibiotic prescribing continues to be an issue in the United States (US) despite a decline in prescription rate overall. Acute gastroenteritis (AGE) often requires medical attention but is generally managed conservatively. We describe antibiotic use among children with AGE in two healthcare settings.Clostridioides difficile detections were excluded due to high colonization rates in this group. A 20-point modified Vesikari score was calculated to categorize AGE severity .Children aged < 5 years with AGE were enrolled from December 2011-August 2021 at 7 New Vaccine Surveillance Network (NVSN) sites across the US. Inpatients (IP) and emergency department (ED) patients with diarrhea (≥3 episodes) and/or vomiting (≥1 episode) within 24 hours, without history of noninfectious diarrhea or immunodeficiency were included. Stools were collected within 10 days of symptom onset and tested for enteric pathogens by validated molecular methods. Antibiotic use during AGE visit was collected via medical chart abstraction; antibiotic use was mostly empiric and not driven by test results. Children < 2 years with Enteropathogenic E. coli .Among 21,194 enrolled children , median IP age was 14 months (IQR 5-28) and ED age 18 months (IQR 9-32). Most were male (IP and ED=54%) and white (IP=61% and ED=43%). Antibiotic use for AGE was noted in 994 (26%) IP and 765 (4%) ED cases; the majority were infants  and had severe AGE . Fever was more common among antibiotic recipients versus non-recipients . Antibiotic use across study years was low in ED (2-10%) but higher in IP (19-31%) . Among tested antibiotic recipients, the predominant pathogens detected were norovirus  and rotavirus  without bacterial codetection, and Overall antibiotic use for AGE among enrolled children was low but was higher among IP, especially in infants and those with severe AGE. Antibiotic use for viral AGE in both ED and IP settings highlights the need to improve antibiotic stewardship among children with AGE.Rangaraj Selvarangan, BVSc, PhD, D(ABMM), FIDSA, FAAM, Abbott: Honoraria|Altona Diagnostics: Grant/Research Support|Baebies Inc: Advisor/Consultant|BioMerieux: Advisor/Consultant|BioMerieux: Grant/Research Support|Bio-Rad: Grant/Research Support|Cepheid: Grant/Research Support|GSK: Advisor/Consultant|Hologic: Grant/Research Support|Lab Simply: Advisor/Consultant|Luminex: Grant/Research Support Christopher J. Harrison, MD, GSK: Grant/Research Support|Pfizer: Grant/Research Support Natasha B. Halasa, MD, MPH, Merck: Grant/Research Support|Quidell: Grant/Research Support|Quidell: donation of kits|Sanofi: Grant/Research Support|Sanofi: vaccine support Geoffrey A. Weinberg, MD, Merck & Co: Honoraria Janet A. Englund, MD, Ark Biopharma: Advisor/Consultant|AstraZeneca: Advisor/Consultant|AstraZeneca: Grant/Research Support|GlaxoSmithKline: Grant/Research Support|Meissa Vaccines: Advisor/Consultant|Merck: Grant/Research Support|Moderna: Advisor/Consultant|Moderna: Grant/Research Support|Pfizer: Advisor/Consultant|Pfizer: Grant/Research Support|Sanofi Pasteur: Advisor/Consultant Mary A. Staat, MD, MPH, CDC: Grant/Research Support|Cepheid: Grant/Research Support|Merck: Grant/Research Support|NIH: Grant/Research Support|Pfizer: Grant/Research Support|Up-To-Date: Honoraria John V. Williams, MD, Merck: Grant/Research Support|Quidel: Board Member Marian G. Michaels, MD, MPH, Merck: Grant/Research Support|Viracor: Grant/Research Support


The elimination of Ringer's solution is severely depressed during general anaesthesia, but the degree to which this continues postoperatively is poorly established.−1 was administered over 60 min in 12 patients undergoing laparoscopic cholecystectomy. Population kinetic analysis was performed based on repeated measurements of blood haemoglobin concentration and urinary excretion over 240 min regardless of when the operations were finished. The analysis contrasted the periods before and after awakening from general anaesthesia and compared them with data from 18 volunteers who received the same fluid at the same rate.An intravenous infusion of Ringer's acetate solution 20 ml kgk21) and the rate constant for urinary excretion (k10) were significantly higher postoperatively than during the surgical period. Computer simulations indicated that urinary excretion after surgery was almost restored to the rate found in the volunteers. In contrast, the redistribution of fluid from the extravascular space to the plasma, which was almost nil during the surgery, showed only limited recovery during the postoperative phase, and was only approximately 10% of the flow rate found in the volunteers. The combination of nearly normalised urinary excretion and lack of adequate return of distributed fluid to the plasma promoted postoperative hypovolaemia.Patients were monitored for approximately 2 h after awakening from general anaesthesia. The rate constant for redistribution of fluid from the extravascular space to the plasma (The kinetic analysis indicates that plasma volume support should be given during the first 2 h after laparoscopic cholecystectomy. Holte and co-workersThe aim of the present study was to contrast the turnover of fluid during the first 2 h after laparoscopic cholecystectomy against the fluid kinetics measured during the preceding period of anaesthesia and surgery. The results were compared with infusion experiments performed in conscious volunteers. The hypothesis was that normal fluid kinetics would only be partially restored in the immediate postoperative period because of the residual effects of the anaesthetics.,This report is a retrospective analysis based on data derived from three published prospective studies where data were collected in the same way. In the first study, plasma dilution was measured as a surrogate for plasma volume expansion when Ringer's acetate solution was administered to 12 patients during elective laparoscopic cholecystectomy.The protocols for the original studies were approved by the Ethics Committee of Huddinge University Hospital , and each subject provided informed written consent to participate. Eleven of the 12 patients had been included in the first study, the female volunteers in the second, and the male volunteers in the third.The patients were allowed to have a light morning breakfast before the study. No premedication was given. With the subject in the supine position, a cannula was placed in the antecubital vein of each arm, one for blood sampling and the other for fluid infusion. General anaesthesia was induced using propofol, rocuronium, and remifentanil, and maintained with remifentanil and sevoflurane as described previously.−1 was given at a constant rate over 60 min via an infusion pump. Ringer's solution has the following ionic content: Na+ 130, K+ 4, Ca2+ 2, Mg2+ 1, Cl− 110, and acetate 30 mmol L−1. The osmolality is approximately 270 mOsm kg−1. Monitoring included pulse oximetry and electrocardiography. Noninvasive arterial pressure was measured in the arm not used for fluid infusion by an automatic device. The excreted urine was measured every 15 min via an indwelling catheter, which had been inserted into the bladder after the induction of anaesthesia but before the infusion started. No other fluid was given during the experiment.When surgery started, an intravenous infusion of acetated Ringer's solution 20 ml kg−1 of the same fluid over 45 min (n=14) or 80 min (n=4). Selection was made from a larger cohort to achieve the same average infusion rate as in the patients. Urine was voided spontaneously halfway through and at the end of each experiment.The volunteers were given 25 ml kgVenous blood samples, 3 ml each, were collected every 5 min during the first 90 min and every 10 min during the subsequent 150 min. All samples were analysed for blood haemoglobin (Hb) concentration at the same hospital's certified clinical chemistry laboratory, with a coefficient of variation (CV) of 1–2%.k12, k21, and k10) and one scaling factor between dilution and volume  was fitted to the dependent variables .A two-volume kinetic model with three rate constants (Ro into the plasma (Vc) from which distribution occurs to (k12) and is redistributed from (k21) an extravascular space (Vt). Elimination from Vc occurs by urinary excretion (k10). All flow rates are proportional by one of these rate constants  to the volume expansion of the respective body fluid space  whereas expanded volumes are shown in lower-case letters (vc and vt), and U is the urinary excretion. Hence, the volume expansion of the central fluid space is given by (vc–Vc).Baseline volumes are given in capital letters  to which covariates were added to evaluate possible changes in kinetics during the postoperative period, and any difference from the volunteers. The criterion for accepting ‘postoperative’ or ‘volunteer’ as covariate in the model required that its inclusion reduced the –2 LL (log likelihood) for the model by >6.6 points (P<0.01).The fluid volume kinetics during general anaesthesia and surgery are described by the parameters in the basic model  in the 30 infusion experiments using the Phoenix 8.3.4 software for non-linear mixed effects  with the First-Order Conditional Estimation Extended Least Squares (FOCE ELS) as a search routine. The sandwich method was used as the variance estimator, as it is robust for covariance misspecification. The programme derived The parameters ultimately reported represented a ‘Full block model’, in which the inter-individual variability of the fixed effects and their covariates (theta:s) are supplemented by the variances and the covariance variability of the random effects (eta:s). A full block model generates the most precise simulations.Goodness-of-fit and performance of the final model were evaluated by predictive checks, residual plots, and plots of the conditional weighted residuals (CWRES).Simulations and plots were performed by inserting the optimal parameter values derived by the analysis into the same kinetic model programmed in MATLAB R2018b .sd), and changes were evaluated using paired t test. Kinetic parameters were reported as the best estimate and 95% CI according to the output from the Phoenix software. The significance levels for inclusion of the covariates were taken from the Phoenix software.Data showing a normal distribution were reported as the mean and standard deviation  who underwent laparoscopic cholecystectomy. They were 43 (11) yr old and weighed 75 (11) kg.The 18 control subjects were volunteers . They were 29 (7) yr old and weighed 70 (13) kg.vs 1.32 (0.35) L, which were administered at rates of 25 (4) and 26 (7) ml min−1, respectively.The average infused volumes were 1.51 (0.21) Patients were monitored for 90 (33) min during ongoing surgery and for an additional 140 (33) min after general anaesthesia had been reversed.Kinetic analysis was performed on all 812 data points on a single occasion. Data on urinary excretion were available on 131 occasions .Three different sets of parameters were derived from the data shown in Compared with the kinetic parameters obtained during general anaesthesia, the parameters representing the postoperative period were characterised by slightly smaller plasma volume expansion in response to the simulated fluid load a. Moreovflow rates showed that the redistribution of fluid from the extravascular space to the plasma was still retarded postoperatively  during the first postoperative 2 h compared with anaesthesia and ongoing surgery.The diuretic response to infused Ringer's solution was much poorer during surgery than in conscious volunteers, but it was almost restored in the early postoperative period after laparoscopic cholecystectomy. The covariance analysis showed a 7-fold increase in the rate constant for urinary excretion (k21) was not normalised after the surgery. This rate constant was almost nil during the surgery, but not even the strong postoperative (hundred-fold) increase in k21 was sufficient to restore the return flow to the plasma.In contrast, the rate constant for the redistribution of fluid from the extravascular space to the plasma  as compared with a restrictive fluid programme (15 ml kg−1).A study in patients undergoing laparoscopic cholecystectomy, where the operating time is usually 1 h, reported improved organ function and recovery when using a liberal fluid regimen (40 ml kgThe Enhanced Recovery After Surgery (ERAS) programme encourages the earliest possible return to oral intake of fluid,Vt), as slow redistribution implies that the extravascular space is not yet able to adequately clear excess fluid.Henriques and CorreiaVc and the interstitial fluid volume to Vt. The flow given by k12 is intended to represent the capillary leakage and k21 primarily the lymphatic return. At steady state, the relationship between k12 and k21 mimics the relationship between Vc and Vt. The fluid kinetic model seems to represent these physiological correlates reasonably well on a whole-body level, provided that the Starling forces are not abruptly changed during an experiment.,The kinetic approach to the analysis and simulation of the distribution of infused fluid has close similarities to drug pharmacokinetics. The model is intended to mimic physiological body fluid spaces, where the plasma corresponds to Vt that does not communicate with the plasma during a normal experimental period of 3–4 h.k21 was almost zero during the surgery. However, the current data set was not sufficiently large to model this second compartment.During surgery, a second elimination compartment might exist alongside k10,Likewise, the arterial pressures probably affect k21.,The poor return of fluid from the extravascular space might be peculiar to the laparoscopic positioning of the patient, as data from thyroid surgery, where the patient rests in the supine position, show a normal Strengths of the study include the fact that the kinetic analysis had a good ability to re-create the measured data, as shown in A kinetic analysis of the fluid kinetics during the first 2 h after laparoscopic cholecystectomy shows that the diuretic response to fluid, which was poor during the surgery, was soon restored to almost the same level as that in volunteers. However, the return flow of distributed fluid to the plasma remained very poor, which promoted peripheral oedema and slight hypovolaemia. Consequently, plasma volume support of modest intensity seems to be warranted in the early postoperative phase after laparoscopic cholecystectomy.Patient recruitment and data collection: JO, RGH.Literature search: JO.Study design, data analysis, and manuscript preparation: RGH.The authors are grateful to the collaborators who helped collect the data from the original studies for creation of this database.10.13039/501100016387Grifols for studies of 20% albumin and is Member of Baxter's IV Fluid Therapy management Advisory Board. JO declares that he has no conflicts of interest.RGH has received a research grant from


Missing FundingIn the original publication , the fun“This research was funded by a subsidy for the maintenance and development of research potential BKM-538/RAU1/2022 (02/040/BKM22/1031) granted by the Polish Ministry of Science and Higher Education (M.K.K), the Polish National Science Center grant 2018/29/B/ST7/02550 (R.J.), the European Union through the European Social Fund grant POWR.03.02.00-00-I029 (P.K.), the NSF/DMS Rapid Collaborative grant NSF/DMS-2030577 to Marek Kimmel and Simon Tavaré (M.K.), and the Polish National Science Center grant 2021/41/B/NZ2/04134 to Marek Kimmel (M.K.).”Text CorrectionThere was an error in the original publication. Practical computations of the SFS based on Tung–Durrett model were carried out using notation A correction has been made to Materials and Methods, SFS under Birth-and-Death Process Model with Mutation and Selection, paragraph 1:0 < λ1, then the site frequency spectrum approximately follows the power curve, in our notationT(x) = ∑ξ>xS(ξ) follows the lawα = λ0/λ1.Under these hypotheses, if the fitnesses of the two types are λThe authors state that the scientific conclusions are unaffected. These corrections were approved by the Academic Editor. The original publication has also been updated.


Prevention of surgical site infections (SSIs) involves implementation of numerous steps including ultraclean air in the operating room (OR). Despite all efforts, particles in the room air may exist and some of these particles may be live pathogens that can potentially cause subsequent SSI. This prospective study aimed to determine and compare the nature and quantity of microbes in the operating room, as detected from the inlet flow of an ultraviolet filtration unit, and the efficacy of the unit to remove particles and creating clean room air (the outlet flow).This prospective study was conducted at a single institution, where primary total joint arthroplasty and spine surgeries were performed. The OR was fitted with a positive ventilation system. In addition, a filtration unit with a crystalline ultraviolet unit (C-UVC) was placed in the OR. The inflow and outflow air from the unit was sampled using specialized swabs at the beginning and at conclusion of each procedure. Additional surgical-related variables were also recorded at each time of sampling. Swabs were processed for culture and Next-Generation Sequencing.Operating room floor planThe filtration unit with ultraviolet-C light is represented with a red rectangle and doors (source of contamination) are represented by dark gray rectangles.Air flow and air swabs collection.The mean length of the surgical procedures sampled was 68 ± 13 minutes. Overall, 19 out of 200 (9.5%) swabs isolated microorganisms. Inflow swabs were positive at a higher rate , compared to the outcoming air swabs. A wide variety of Gram-positive, Gram-negative, anaerobic bacteria, and fungi (only in the inflow swabs) were isolated. The detection of microorganisms was also higher in surgical procedures with a higher number of door openings .Surgical-related variables collected at air swab sampling.Microorganisms are present in the operating room air. A specialized filtration unit with a C-UVC light was effective in filtering these microorganisms in the majority of cases.Javad Parvizi, MD, FRCS, 3M: Grant/Research Support|Acumed, LLC: Stocks/Bonds|Aesculap: Grant/Research Support|Alphaeon: Stocks/Bonds|AO Spine: Stocks/Bonds|Becton Dickenson: Advisor/Consultant|Biomet: Grant/Research Support|Cardinal Health: Advisor/Consultant|Cempra: Grant/Research Support|CeramTec: Grant/Research Support|Ceribell: Stocks/Bonds|Coracoid: Stocks/Bonds|Corentec: Advisor/Consultant|Datatrace: Grant/Research Support|DePuy: Grant/Research Support|Elsevier: Grant/Research Support|Elute: Stocks/Bonds|Ethicon: Advisor/Consultant|Hip Innovation Technology: Stocks/Bonds|Illuminus: Stocks/Bonds|Integra: Grant/Research Support|Intellijoint: Stocks/Bonds|Jaypee Publishers: Grant/Research Support|KCI / 3M (Acelity): Advisor/Consultant|Lima: Grant/Research Support|MicroGenDx: Advisor/Consultant|Molecular Surface Technologies: Stocks/Bonds|Myoscience: Grant/Research Support|Nanooxygenic: Stocks/Bonds|National Institutes of Health (NIAMS & NICHD): Grant/Research Support|NDRI: Grant/Research Support|Novartis: Grant/Research Support|OREF: Grant/Research Support|Orthospace: Grant/Research Support|Osteal: Stocks/Bonds|Parvizi Surgical Innovations and Subsidiaries: Stocks/Bonds|Peptilogic: Stocks/Bonds|Peptilogics: Advisor/Consultant|Pfizer: Grant/Research Support|PRN-Veterinary: Grant/Research Support|Rotation Medical: Grant/Research Support|Simplify Medical: Grant/Research Support|SLACK Incorporated: Grant/Research Support|Smith & Nephew: Grant/Research Support|Sonata: Stocks/Bonds|Stelkast: Grant/Research Support|Stryker: Grant/Research Support|Synthes: Grant/Research Support|Tenor: Advisor/Consultant|TissueGene: Grant/Research Support|Tornier: Grant/Research Support|Wolters Kluwer Health - Lippincott Williams & Wilkins: Grant/Research Support|Zimmer Biomet: Advisor/Consultant|Zimmer Biomet: Grant/Research Support


Drilling cuttings from the rock formation generated during the drilling process are generally smashed to fine particles through hydraulic cutting and grinding using a drilling tool, and then are mixed with the drilling fluid during circulation. However, some of these particles are too small and light to be effectively removed from the drilling fluid via solids-control equipment. These small and light solids are referred to as low gravity solids (LGSs). This work aimed to investigate the effect of LGSs on the performance of oil-based drilling fluid (OBDF), such as the rheological properties, high-temperature and high-pressure filtration loss, emulsion stability, and filter cake quality. The results show that when the content of LGSs reached or even exceeded the solid capacity limit of the OBDF, the rheological parameters including the plastic viscosity, gel strength, and thixotropy of OBDF increased significantly. Furthermore, the filtration of OBDF increases, the filter cake becomes thicker, the friction resistance becomes larger, and the stability of emulsion of OBDF also decreases significantly when the concentration of LGSs reached the solid capacity limit of OBDF (6–9 wt% commonly). It was also found that LGSs with a smaller particle size had a more pronounced negative impact on the drilling fluid performance. This work provides guidance for understanding the impact mechanism of LGSs on drilling fluid performance and regulating the performance of OBDF. Oil-based drilling fluid (OBDF) refers to a non-aqueous solvent drilling fluid that uses oil  as the continuous phase, as opposed to water-based drilling fluid ,2,3. OBDsV is the volume of rock fragments entering the OBDF, φ is the average porosity of the formation, r is the bit radius, and ROP is an abbreviation for the rate of penetration. Usually, up to 100,000 pounds of rock debris could be carried with the drilling fluid every day.During the drilling process, drilling cuttings composed of rocks and low-quality clay are mixed into OBDF. In addition, the commercial solids  intentionally added to OBDF, drilling cuttings represent a significant source of solid particles. The volume of rock debris or drilling solids removed from the borehole is enormous and can be estimated using the following formula:The primary function of a drilling fluid is to carry the drilling cuttings generated by the drill bit during the drilling process. This is achieved via circulation, where the drilling fluid carries the cuttings to the surface, and they are subsequently removed using solid-control equipment. This ensures continuous drilling and the advancement of the wellbore ,6. FurthHowever, due to the high preparation cost of OBDF, the method of multi-well circulation is applied to reduce the overall cost of drilling fluid. Recycling drilling fluid and improper solid-control methods could lead to a gradual increase in the LGSs’ content. Because it is impossible to entirely remove all of the drilling particles from the drilling fluid, they are considered persistent pollutants in the fluid system, which highlights the significance of this study. The degree of pollution caused by these drilling solids depends largely on their content, particle size, and type . Up to nSince the LGSs are an inevitable product during the drilling process, understanding the effect of LGSs on the performance of OBDF is crucial for adjusting the drilling fluid properties and mitigating the deterioration of the drilling fluid performance. Furthermore, it is necessary to monitor closely the content of LGS in OBDF and maintain the drilling fluid in a timely manner during the drilling process.With this in mind, the objective of this study is to evaluate the impact of LGSs on various important properties of OBDF, including rheology, high-temperature and high-pressure (HTHP) filtration, emulsion stability, and filter cake quality. Firstly, a new method was designed to separate LGSs from waste OBDF. Then, the impact of LGSs on the rheological properties of OBDF is evaluated through adding the separated LGSs into the freshly prepared OBDF. Differently from previous papers, this study further investigated the influence of LGS on the rheological properties of drilling fluids via steady-state shear experiments, the recovery rate of drilling fluid structures at low shear rates, and the thixotropy ring method. Furthermore, to gain a more comprehensive understanding of the impact of LGS on filtration, emulsion stability, and the other properties of drilling fluids, the filtration test, mud cake property, and emulsion stability test were performed.3) and the weighting material (4.2 g/cm3) in the drilling fluid. An airflow classifier  was further used to classify LGSs based on their particle sizes. In the airflow classifier, LGSs were subject to the impact and disturbance of the airflow. Due to the different particle sizes of LGSs, the motion state of LGSs changed accordingly. In this work, LGSs were divided into three categories according to their median particle size (D50), i.e., LGS-14.5, LGS-9.1, and LGS-6.6.2/kg, 1185.754 m2/kg, and 1579.346 m2/kg, respectively, which is consistent with the results of the particle size reduction.As shown in −1) is positively correlated with the concentration of LGSs  of 17.23 Pa, 27.66 Pa, 44.70 Pa, 86.56 Pa, 145.82 Pa, and 198.62 Pa for drilling fluids with the LGSs’ concentrations of 0 wt%, 3 wt%, 6 wt%, 9 wt%, 12 wt%, and 15 wt%, respectively. The gel strength of the drilling fluid is also positively correlated with the amount of LGSs added. Similarly, the fitted plastic viscosity values (ηp) of 97.44 mPa·s, 126.01 mPa·s, 165.73 mPa·s, 175.99 mPa·s, 212.00 mPa·s, and 256.06 mPa·s for drilling fluids with the LGSs’ concentrations of 0 wt%, 3 wt%, 6 wt%, 9 wt%, 12 wt%, and 15 wt%, respectively, indicate a positive correlation between the frictional resistance in the drilling fluid and the amount of LGSs added. In order to reduce the annular circulation pressure loss, reduce the bottom hole back pressure, and increase the mechanical drilling speed, regardless of the drilling fluid density or other requirements of the downhole working conditions, it is better to have a smaller ηp because of the annular circulation pressure loss Δp∞ηp. τ0 is one of the most obvious rheological parameters that determine the fluctuating pressure. A large number of studies abroad have shown that only changing the ratio from 30 Pa to 50 Pa significantly increases the bottom fluctuation pressure, and the annular pressure loss Δp is also proportional to τ0. Therefore, the maximum allowable limit set by foreign countries is 50 Pa. Moreover, research has shown that for the benefit of wellbore purification, the minimum limit of τ0 is 15~20 Pa, and the density of the high-density drilling fluid helps to improve the rock carrying efficiency without requiring too high rheological parameters of the carrier. 0 and ηp with the LGSs’ concentration. It was observed that τ0 satisfied an exponential equation with the LGSs’ concentration, while ηp satisfied a linear equation. This is because the yield stress reflects the interaction force between the particles and the drilling fluid treatment agents during laminar flow of fluid. This force significantly increases with the increase in solid particles, resulting in a geometrically amplified strength of the spatial grid structure. The plastic viscosity reflects the internal friction between particles, droplets, and liquids in the drilling fluid. It is mainly related to the solid content or particle number. The viscosity of the suspension in consistent units is given via the Krieger–Dougherty equation  is the intrinsic viscosity of the medium.As shown in Thixotropy is also an important rheological parameter of drilling fluid. Thixotropy is the structural degradation caused by the breakage of flocculated or interconnected particles during shearing. When the shear stress is eliminated, the microstructure of drilling fluid will be reconstructed and ultimately restored to its original state ,22. Dril−1, indicating the presence of a high strength gel structure in drilling fluid. However, when the shear rate increased to 100 s−1, the viscosity of the drilling fluid gradually decreased and stabilized, indicating that the structure in the drilling fluid was damaged and changed to a sol state. Furthermore, when the shear rate restored to 0.1 s−1, the viscosity of the drilling fluid gradually recovered to a high viscosity, indicating a gradual restoration of the drilling fluid structure. In addition, as the LGSs’ content increased, the viscosity of the drilling fluid also increased. When the LGSs’ content in the drilling fluid was increased from 0 wt% to 15 wt%, the viscosity of the drilling fluid increased by nearly 100 times at 0.1 s−1 and nearly 10 times at 100 s−1. Moreover, when the viscosity of the drilling fluid was decreased again to a low shear rate, the recovery rate of viscosity was faster, indicating that the introduction of LGSs enhanced the structural recovery ability of the drilling fluid at low shear rates. The results of structure recovery show that . When LGSs was added, the viscosity increased with the size of the LGSs’ particles added; however, when the shear rate was increased to 100 s−1, there was minimal variation in viscosity among drilling fluids with various LGSs’ particle sizes. The thixotropic loop method was used to evaluate the impact of the LGSs’ size on the thixotropy of OBDF, as depicted in Furthermore, the interaction between particles is weakened with the increase in shear rate. This phenomenon is evident in As shown in In addition, emulsion stability (ES) is also one of the most important properties of OBDF. The effect of the LGSs’ content on ES was investigated, as shown in As shown in In summary, a novel method was developed to separate the LGSs from waste OBDF. Then, the separated LGSs were added to newly prepared OBDF to investigate the effects of the LGSs’ concentration and particle sizes on the performance of the OBDF. The results of the rheological test indicated that when the concentration of LGS in OBDF exceeded 6 wt%, LGSs formed a strong spatial grid structure in the drilling fluid, resulting in increased yield stress, plastic viscosity, and the thixotropy of OBDF. This relatively strong gel structure is detrimental to drilling operations. Furthermore, when the solid capacity limit of OBDF is exceeded, adding excessive LGSs results in thickened mud cakes, increased friction resistance, decreased emulsion stability, and increased filtrate loss of drilling fluid. Moreover, the particle size of the LGSs plays a significant role, where smaller particle sizes have a more pronounced negative impact on the drilling fluid performance. A lower solid capacity limit for the LGS with smaller particle sizes can be observed. The impact of LGSs on the performance of drilling fluids exhibits both a “cumulative effect” and a “size effect”. The “cumulative effect” and “size effect” can be summarized as an exponential relationship between the degree of deterioration of the drilling fluid performance and the content of LGS, and this deterioration is related to the particle size of LGS. The smaller the particle size, the more significant negative impact on the drilling fluid performance. We believe that these findings provides guidance for understanding the impact mechanism of LGSs on the drilling fluid performance and regulating the drilling fluid performance. Moreover, considering the negative impact of LGS on the drilling fluid performance, drilling fluid engineers should pay more attention to the removal of LGS and reasonably use solid phase control equipment to reduce the LGS content in the drilling fluid as much as possible. At the same time, when it is found that the LGS content is too high, additional drilling fluid dilution with a new configuration is used to reduce the LGS concentration.2, AR), calcium oxide , petroleum ether (AR), methylene iodide , and mineral oil were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., Shanghai, China. The weighting agent, barite, was obtained from Hubei Hanc New-Technology Co., Ltd., Jingzhou, China.The primary emulsifier, secondary emulsifier, wetting agent, organic clay, and filtrate reducer were all provided by China National Offshore Oil Corporation (CNOOC) Co., Ltd., Tianjin, China. Calcium chloride  with stirring at 1000 rpm, and then centrifuged at 2000 rpm for 5 min. The density of LGSs is usually considered to be 2.6 g/cm3, while the density of barite used in drilling fluids is usually 4.2 g/cm3. According to the principle of settlement, low-density solid particles tend to float upwards, while heavier materials settle down. Finally, the LGSs were collected by filtering the upper liquid. The separation process of the LGSs is illustrated in The waste OBDF sample was diluted three times with mineral oil and subjected to centrifugation for 10 min at 10,000 rpm using a high-speed centrifuge  to collect the sediment at the bottom. Then, the collected sediment was transferred to a Soxhlet extractor and subjected to extraction for 12 h to remove the mineral oil and treatment agents adsorbed on the solids’ surface. The solids in the waste OBDF were obtained by drying the sample at 70 °C until a constant weight was achieved. The obtained solids were subsequently dispersed into diiodomethane (density: 3.32 g/cm50), the LGSs with different particles were marked as LGS-6.6, LGS-9.1, and LGS-14.5.After drying the separated LGSs, an airflow classifier was used to separate the LGSs into three different particle sizes. According to the median particle size :τ is shear stress, γ is shear rate, and η is viscosity. The τ as a function of γ was fitted according to the Bingham Model (Equation (4)) to obtain the yield stress (τ0) and plastic viscosity (pη).The rheological properties of the OBDF were tested using a MARS60 rheometer (Thermo Fisher Scientific Inc. Germany) equipped with a parallel plate geometry . All experiments were carried out at 25 °C. The steady shearing measurements were performed to record the viscosity at shear rates ranging from 10−1) and high shear rates (100 s−1). For the thixotropic loop method, the applied shear rate was increased from 0 to 300 s−1 within 60 s and kept at 300 s−1 for 10 s. Then, the shear rate was decreased within the same range for the same duration of 60 s. The corresponding shear stress was monitored for each imposed shear rate.The thixotropic behavior was evaluated via the three-stage rheological test and the thixotropic loop method. The three-stage rheological test involved measuring the change in viscosity over time when applying the alternating low :τ0 and the pressure loss in the pipeline ΔThe relationship between dynamic shear force R1 is the outer radius of the drill pipe, R2 is the wellbore radius, and mR is the distance from the maximum flow velocity point in the annular to the axial diameter of the wellbore. mR can be calculated from Equation (9):The pressure loss in the annulus can be obtained from Equation (8):HTHP) of the OBDF at 120 °C was measured using an HPHT instrument . As per the API testing standard for testing drilling fluids (API 13-B), the filtration period was set to 30 min, and the differential pressure was kept at 3.5 MPa.The high-temperature and high-pressure filtration . Firstly, a mud cake obtained after the filtrate loss test was placed on a horizontal platform, and a stainless steel cylinder with a diameter of 20 mm and a length of 60 mm was placed on the surface of the mud cake. The mud cake sticking coefficient tester rotated the platform and measured the rotation angle at which the steel column started to slide after overcoming the frictional forces. Initially, the steel column remained stationary due to the friction between the mud cake and the stainless steel cylinder. However, when the rotation angle of the platform reached a critical value, the steel column began to slide downwards under the action of gravity. Finally, the sticking coefficient was obtained by calculating the tangent value of the critical angle of the platform.The emulsion stability (ES) of the OBDF was investigated via an electrical stability measurement  with an electrode distance of 1.55 ± 0.04 mm at 25 °C. The electrode probe was placed in the middle of OBDF, and the emulsion-breaking voltage was accorded.


Anaesthesia plays an important role in diagnosis procedures and the treatment and pain management of oncological patients. However, studies suggest that anaesthetic drugs may increase the risk of tumour dissemination in the perioperative period by directly and indirectly suppressing the immune system, which is primarily responsible for controlling tumour growth. Science faces the challenge of understanding their influence on the immune system to develop anaesthetic strategies that assure greater immune stability in patients with cancer or other immune-compromising diseases.Throughout the course of oncological disease, the majority of patients require surgical, anaesthetic and analgesic intervention. However, during the perioperative period, anaesthetic agents and techniques, surgical tissue trauma, adjuvant drugs for local pain and inflammation and other non-pharmacological factors, such as blood transfusions, hydration, temperature and nutrition, may influence the prognosis of the disease. These factors significantly impact the oncologic patient’s immune response, which is the primary barrier to tumour progress, promoting a window of vulnerability for its dissemination and recurrence. More research is required to ascertain which anaesthetics and techniques have immunoprotective and anti-tumour effects, which will contribute to developing novel anaesthetic strategies in veterinary medicine. The growing attention paid to animal health  and the Considering human and animal studies, the main risk of tumour dissemination occurs in the perioperative period ,16, duriAccording to this knowledge, this review aims to debate the influence of anaesthetic drugs on the immune system and tumour cells based on retrospective studies in humans and animals.This review has been conducted to describe the outcomes of relevant experimental studies. A search strategy of the PubMed database was conducted to find the qualifying articles. We applied the following keywords: “anaesthetic strategies in oncological surgery”; “anaesthesia and cancer”; “anaesthesia and cancer recurrence and metastasis”; “veterinary anaesthesia oncology”; “cancer in anaesthesia dogs”; “anaesthesia and cancer recurrence and metastasis in dogs”; “regional anaesthesia in animals”; “total intravenous anaesthesia in small animals”; “intravenous anaesthetics and tumour spread”; “volatile anaesthetics and tumour spread”; “surgery and immune system”; “surgery and tumour dissemination”; “perioperative period and tumour spread”; “cancer and immune system”. The search terms determined were full text, ten years, randomised controlled trial, clinical trial, meta-analysis, review and veterinary anaesthesia.The tumour growth and spread process is influenced by the tumour’s characteristics but mainly by the host’s immune system . The sciIn veterinary and human searches, a high count and ratio of circulating leukocytes  ,32, as wConsidering the immune system as a main player in the body’s defence against tumour growth, some human and animal studies suggest a new approach to minimising the impact on the immune cells of several suppressive factors such as stress, pain, anaesthesia and surgery ,44,45. AExcisional tumour surgery is the main approach for treating primary solid tumours  and the First, the surgical manipulation of neoplastic tissue promotes the release of neoplastic cells into the blood and lymphatic system , where tSecondly, the microenvironment of the damaged tissue in the area of the surgical procedure changes, and minutes after the incision, a proportional local and systemic stress response is generated . PhysiolConsidering the inflammatory, immunosuppressive and tumour-spreading effects of surgery trauma, the potential advantages of minimally invasive surgery (MIS) over conventional open surgery have been discussed in human and animal studies, but with inconsistent results ,64. MIS Lastly, tumour dormancy is a condition characterised by the presence of tumour cells in the absence of evidence of oncological disease . This phEven though some biological factors may paradoxically enhance the proliferation and dissemination of metastatic cells, surgical tumour removal provides an evident benefit in preventing the growth and spread of cancer . FurtherOncological diseases often require the use of anaesthetic and analgesic agents during diagnostic ,79, therVolatile anaesthetics, such as isuflurane and sevoflurane, are halogenated agents used to induce and maintain general anaesthesia through the respiratory system  as well Isuflurane is an inhaled anaesthetic agent commonly used in veterinary medicine that produces a state of unconsciousness due to its action on the central nervous system and without analgesic properties . Like otSevoflurane is an inhaled anaesthetic agent used to induce and maintain anaesthesia . It prodPropofol, alphaxalone and ketamine are all commonly used for the induction, co-induction and maintenance of anaesthesia ,127,128.Propofol  is one of the intravenous anaesthetic agents routinely used in veterinary clinical practice for the induction and maintenance of general anaesthesia, with affinity for γ-aminobutyric acid (GABA) receptors and producing sedative and hypnotic effects . They poTotal intravenous anaesthesia (TIVA) is defined as the induction and maintenance of anaesthesia achieved exclusively by intravenous (IV) drugs . This apAlfaxalone  is a neurosteroid anaesthetic that interacts with the gamma aminobutyric acid A receptor, producing anaesthesia and muscle relaxation . MoreoveKetamine is a phenylcyclohexylamine derivative and antagonist of the excitatory neurotransmitter N-methyl-D-aspartate (NMDA) receptors . It is wThe influence of surgery and anaesthetic agents on the immune system and tumour cells is still uncertain. Despite several studies, the controversial outcomes remain a fact and can be attributed to distinct experimental conditions, such as in vitro or in vivo research, the type of cell line, surgical technique, and anaesthetic agents and their concentration. Moreover, we know that the perioperative period is a time when the body is more susceptible to developing a compromised immune response and, consequently, an increased risk of tumour dissemination. However, this time can be seen as an opportunity to control or minimise the impact of anaesthetic and surgical effects. After reviewing the above studies, propofol-based anaesthesia appears to have immunoprotective and anti-tumour effects and has been shown to decrease surgical stress, immunosuppression and angiogenesis. On the other hand, inhaled anaesthetics have been associated with a higher rate of tumour recurrence and metastasis because of their capacity to induce pro-inflammatory activity and decrease immune function. As well, minimally invasive surgery, when compared with open surgery, shows major tissue protection and minor stress responses. However, it has been linked to prolonged surgical and anaesthetic times.Through the new anaesthesia and surgery strategy, the perioperative period is already recognised as a window of opportunity for minimising their side effects. To draw definitive conclusions about the outcomes of surgery and anaesthesia in oncology patients, additional clinical trials are required.


Ceftobiprole is being developed for the treatment of patients with S. aureus bacteremia (SAB). Pharmacokinetic-pharmacodynamic (PK-PD) relationships for efficacy were evaluated using data from ceftobiprole-treated patients with SAB from the ERADICATE Phase 3 study .Ceftobiprole medocaril is an intravenously (IV) administered cephalosporin prodrug that is rapidly converted S. aureus and PK data were considered. On Days 1-8, patients received ceftobiprole 500 mg IV every 6 hours followed by 500 mg IV every 8 hours on Day 9 and onwards with dose adjustment for renal impairment. Using a population PK model and PK data from patients, ceftobiprole free-drug plasma %T >MIC on Days 1 and 10 were determined. Univariable relationships between the dichotomous or time-to-event efficacy endpoints summarized in Table 1 and each of Days 1 or 10 free-drug plasma %T >MIC and baseline MIC were evaluated.Ceftobiprole-treated patients in the modified Intent-to-Treat (mITT) populations of Cohorts 1 and 2  with an MIC value for the baseline Table 2. As shown in Table 3, nearly all patients achieved Days 1 and 10 ceftobiprole free-drug plasma %T > MIC ≥ 97.9%. Given this distribution, the evaluation of associations with efficacy endpoints was limited. There was no substantial evidence of relationships between either increased free-drug plasma %T > MIC or reduced baseline MIC and increased efficacy among the endpoints assessed. All patients achieved free-drug plasma %T > MIC that exceeded the non-clinical free-drug plasma %T > MIC target of 24.7% associated with a 1-log10 CFU reduction from baseline for S. aureus.A summary of response for efficacy endpoints among the 172 evaluable patients with SAB is provided in Results of these analyses did not demonstrate relationships between either ceftobiprole free-drug plasma %T > MIC or baseline MIC and response. Given the favorable Phase 3 ERADICATE study findings of ceftobiprole non-inferiority relative to daptomycin, these data suggested that ceftobiprole exposures were efficacious and provide support for the ceftobiprole dosing regimens evaluated.Sujata M. Bhavnani, PharmD; MS; FIDSA, Adagio Therapeutics, Inc.: Grant/Research Support|Albany Medical Center: Grant/Research Support|Amplyx Pharmaceuticals, Inc.: Grant/Research Support|AN2 Therapeutics: Grant/Research Support|Antabio SAS: Grant/Research Support|Arcutis Biotherapeutics, Inc.: Grant/Research Support|B. Braun Medical Inc.: Grant/Research Support|Basilea Pharmaceutica: Grant/Research Support|BioFire Diagnostics LLC: Grant/Research Support|Boston Pharmaceuticals: Grant/Research Support|Cidara Therapeutics Inc.: Grant/Research Support|Cipla USA: Grant/Research Support|Crestone Inc.: Grant/Research Support|CXC: Grant/Research Support|Debiopharm International SA: Grant/Research Support|Entasis Therapeutics: Grant/Research Support|Genentech: Grant/Research Support|GlaxoSmithKline: Grant/Research Support|Hoffmann-La Roche: Grant/Research Support|ICPD: Ownership Interest|Inotrem: Grant/Research Support|Insmed Inc.: Grant/Research Support|Iterum Therapeutics Limited: Grant/Research Support|Kaizen Bioscience, Co.: Grant/Research Support|KBP Biosciences USA: Grant/Research Support|Matinas Biopharma: Grant/Research Support|Meiji Seika Pharma Co., Ltd.: Grant/Research Support|Melinta Therapeutics: Grant/Research Support|Menarini Ricerche S.p.A.: Grant/Research Support|Mutabilis: Grant/Research Support|Nabriva Therapeutics AG: Grant/Research Support|Paratek Pharmaceuticals, Inc.: Grant/Research Support|Qpex Biopharma: Grant/Research Support|Sfunga Therapeutics: Grant/Research Support|Spero Therapeutics: Grant/Research Support|Suzhou Sinovent Pharmaceuticals Co.: Grant/Research Support|Theravance: Grant/Research Support|tranScrip Partners: Grant/Research Support|University of Wisconsin: Grant/Research Support|Utility Therapeutics: Grant/Research Support|ValanBio Therapeutics Inc.: Grant/Research Support|VenatoRx: Grant/Research Support Jeffrey P. Hammel, MS, Adagio Therapeutics, Inc.: Grant/Research Support|Albany Medical Center: Grant/Research Support|Amplyx Pharmaceuticals, Inc.: Grant/Research Support|AN2 Therapeutics: Grant/Research Support|Antabio SAS: Grant/Research Support|Arcutis Biotherapeutics, Inc.: Grant/Research Support|B. Braun Medical Inc.: Grant/Research Support|Basilea Pharmaceutica: Grant/Research Support|BioFire Diagnostics LLC: Grant/Research Support|Boston Pharmaceuticals: Grant/Research Support|Cidara Therapeutics Inc.: Grant/Research Support|Cipla USA: Grant/Research Support|Crestone Inc.: Grant/Research Support|CXC: Grant/Research Support|Debiopharm International SA: Grant/Research Support|Entasis Therapeutics: Grant/Research Support|Genentech: Grant/Research Support|GlaxoSmithKline: Grant/Research Support|Hoffmann-La Roche: Grant/Research Support|ICPD: Employee|Inotrem: Grant/Research Support|Insmed Inc.: Grant/Research Support|Iterum Therapeutics Limited: Grant/Research Support|Kaizen Bioscience, Co.: Grant/Research Support|KBP Biosciences USA: Grant/Research Support|Matinas Biopharma: Grant/Research Support|Meiji Seika Pharma Co., Ltd.: Grant/Research Support|Melinta Therapeutics: Grant/Research Support|Menarini Ricerche S.p.A.: Grant/Research Support|Mutabilis: Grant/Research Support|Nabriva Therapeutics AG: Grant/Research Support|Paratek Pharmaceuticals, Inc.: Grant/Research Support|Qpex Biopharma: Grant/Research Support|Sfunga Therapeutics: Grant/Research Support|Spero Therapeutics: Grant/Research Support|Suzhou Sinovent Pharmaceuticals Co.: Grant/Research Support|Theravance: Grant/Research Support|tranScrip Partners: Grant/Research Support|University of Wisconsin: Grant/Research Support|Utility Therapeutics: Grant/Research Support|ValanBio Therapeutics Inc.: Grant/Research Support|VenatoRx: Grant/Research Support Kathryn Liolios, MA, Adagio Therapeutics, Inc.: Grant/Research Support|Albany Medical Center: Grant/Research Support|Amplyx Pharmaceuticals, Inc.: Grant/Research Support|AN2 Therapeutics: Grant/Research Support|Antabio SAS: Grant/Research Support|Arcutis Biotherapeutics, Inc.: Grant/Research Support|B. Braun Medical Inc.: Grant/Research Support|Basilea Pharmaceutica: Grant/Research Support|BioFire Diagnostics LLC: Grant/Research Support|Boston Pharmaceuticals: Grant/Research Support|Cidara Therapeutics Inc.: Grant/Research Support|Cipla USA: Grant/Research Support|Crestone Inc.: Grant/Research Support|CXC: Grant/Research Support|Debiopharm International SA: Grant/Research Support|Entasis Therapeutics: Grant/Research Support|Genentech: Grant/Research Support|GlaxoSmithKline: Grant/Research Support|Hoffmann-La Roche: Grant/Research Support|ICPD: Employee|Inotrem: Grant/Research Support|Insmed Inc.: Grant/Research Support|Iterum Therapeutics Limited: Grant/Research Support|Kaizen Bioscience, Co.: Grant/Research Support|KBP Biosciences USA: Grant/Research Support|Matinas Biopharma: Grant/Research Support|Meiji Seika Pharma Co., Ltd.: Grant/Research Support|Melinta Therapeutics: Grant/Research Support|Menarini Ricerche S.p.A.: Grant/Research Support|Mutabilis: Grant/Research Support|Nabriva Therapeutics AG: Grant/Research Support|Paratek Pharmaceuticals, Inc.: Grant/Research Support|Qpex Biopharma: Grant/Research Support|Sfunga Therapeutics: Grant/Research Support|Spero Therapeutics: Grant/Research Support|Suzhou Sinovent Pharmaceuticals Co.: Grant/Research Support|Theravance: Grant/Research Support|tranScrip Partners: Grant/Research Support|University of Wisconsin: Grant/Research Support|Utility Therapeutics: Grant/Research Support|ValanBio Therapeutics Inc.: Grant/Research Support|VenatoRx: Grant/Research Support Anthony P. Cammarata, M.S., Adagio Therapeutics, Inc.: Grant/Research Support|Albany Medical Center: Grant/Research Support|Amplyx Pharmaceuticals, Inc.: Grant/Research Support|AN2 Therapeutics: Grant/Research Support|Antabio SAS: Grant/Research Support|Arcutis Biotherapeutics, Inc.: Grant/Research Support|B. Braun Medical Inc.: Grant/Research Support|Basilea Pharmaceutica: Grant/Research Support|BioFire Diagnostics LLC: Grant/Research Support|Boston Pharmaceuticals: Grant/Research Support|Cidara Therapeutics Inc.: Grant/Research Support|Cipla USA: Grant/Research Support|Crestone Inc.: Grant/Research Support|CXC: Grant/Research Support|Debiopharm International SA: Grant/Research Support|Entasis Therapeutics: Grant/Research Support|Genentech: Grant/Research Support|GlaxoSmithKline: Grant/Research Support|Hoffmann-La Roche: Grant/Research Support|ICPD: Employee|Inotrem: Grant/Research Support|Insmed Inc.: Grant/Research Support|Iterum Therapeutics Limited: Grant/Research Support|Kaizen Bioscience, Co.: Grant/Research Support|KBP Biosciences USA: Grant/Research Support|Matinas Biopharma: Grant/Research Support|Meiji Seika Pharma Co., Ltd.: Grant/Research Support|Melinta Therapeutics: Grant/Research Support|Menarini Ricerche S.p.A.: Grant/Research Support|Mutabilis: Grant/Research Support|Nabriva Therapeutics AG: Grant/Research Support|Paratek Pharmaceuticals, Inc.: Grant/Research Support|Qpex Biopharma: Grant/Research Support|Sfunga Therapeutics: Grant/Research Support|Spero Therapeutics: Grant/Research Support|Suzhou Sinovent Pharmaceuticals Co.: Grant/Research Support|Theravance: Grant/Research Support|tranScrip Partners: Grant/Research Support|University of Wisconsin: Grant/Research Support|Utility Therapeutics: Grant/Research Support|ValanBio Therapeutics Inc.: Grant/Research Support|VenatoRx: Grant/Research Support Mikael Saulay, MSc, Basilea Pharmaceutica International Ltd, Allschwil, Switzerland: Full time employee|Basilea Pharmaceutica International Ltd, Allschwil, Switzerland: Stocks/Bonds Christopher M. Rubino, PharmD, Adagio Therapeutics, Inc.: Grant/Research Support|Albany Medical Center: Grant/Research Support|Amplyx Pharmaceuticals, Inc.: Grant/Research Support|AN2 Therapeutics: Grant/Research Support|Antabio SAS: Grant/Research Support|Arcutis Biotherapeutics, Inc.: Grant/Research Support|B. Braun Medical Inc.: Grant/Research Support|Basilea Pharmaceutica: Grant/Research Support|BioFire Diagnostics LLC: Grant/Research Support|Boston Pharmaceuticals: Grant/Research Support|Cidara Therapeutics Inc.: Grant/Research Support|Cipla USA: Grant/Research Support|Crestone Inc.: Grant/Research Support|CXC: Grant/Research Support|Debiopharm International SA: Grant/Research Support|Entasis Therapeutics: Grant/Research Support|Genentech: Grant/Research Support|GlaxoSmithKline: Grant/Research Support|Hoffmann-La Roche: Grant/Research Support|ICPD: Ownership Interest|Inotrem: Grant/Research Support|Insmed Inc.: Grant/Research Support|Iterum Therapeutics Limited: Grant/Research Support|Kaizen Bioscience, Co.: Grant/Research Support|KBP Biosciences USA: Grant/Research Support|Matinas Biopharma: Grant/Research Support|Meiji Seika Pharma Co., Ltd.: Grant/Research Support|Melinta Therapeutics: Grant/Research Support|Menarini Ricerche S.p.A.: Grant/Research Support|Mutabilis: Grant/Research Support|Nabriva Therapeutics AG: Grant/Research Support|Paratek Pharmaceuticals, Inc.: Grant/Research Support|Qpex Biopharma: Grant/Research Support|Sfunga Therapeutics: Grant/Research Support|Spero Therapeutics: Grant/Research Support|Suzhou Sinovent Pharmaceuticals Co.: Grant/Research Support|Theravance: Grant/Research Support|tranScrip Partners: Grant/Research Support|University of Wisconsin: Grant/Research Support|Utility Therapeutics: Grant/Research Support|ValanBio Therapeutics Inc.: Grant/Research Support|VenatoRx: Grant/Research Support Marc Engelhardt, MD, Astellas Pharma Global Development, Inc.: Support for the present publication|Basilea Pharmaceutica International Ltd: Employee of Basilea Pharmaceutica International Ltd|Basilea Pharmaceutica International Ltd: Stocks/Bonds Jennifer Smart, PhD, Basilea Pharmaceutica International Ltd, Allschwil, Switzerland: Stocks/Bonds Mark E. Jones, PhD, Astellas Pharma Global Development, Inc: Support for the present publication|Basilea Pharmaceutica International Ltd: Employee of Basilea Pharmaceutica International Ltd|Basilea Pharmaceutica International Ltd: Stocks/Bonds Paul G. Ambrose, PharmD; MS; FIDSA, Adagio Therapeutics, Inc.: Grant/Research Support|Albany Medical Center: Grant/Research Support|Amplyx Pharmaceuticals, Inc.: Grant/Research Support|AN2 Therapeutics: Grant/Research Support|Antabio SAS: Grant/Research Support|Arcutis Biotherapeutics, Inc.: Grant/Research Support|B. Braun Medical Inc.: Grant/Research Support|Basilea Pharmaceutica: Grant/Research Support|BioFire Diagnostics LLC: Grant/Research Support|Boston Pharmaceuticals: Grant/Research Support|Cidara Therapeutics Inc.: Grant/Research Support|Cipla USA: Grant/Research Support|Crestone Inc.: Grant/Research Support|CXC: Grant/Research Support|Debiopharm International SA: Grant/Research Support|Entasis Therapeutics: Grant/Research Support|Genentech: Grant/Research Support|GlaxoSmithKline: Grant/Research Support|Hoffmann-La Roche: Grant/Research Support|ICPD: Ownership Interest|Inotrem: Grant/Research Support|Insmed Inc.: Grant/Research Support|Iterum Therapeutics Limited: Grant/Research Support|Kaizen Bioscience, Co.: Grant/Research Support|KBP Biosciences USA: Grant/Research Support|Matinas Biopharma: Grant/Research Support|Meiji Seika Pharma Co., Ltd.: Grant/Research Support|Melinta Therapeutics: Grant/Research Support|Menarini Ricerche S.p.A.: Grant/Research Support|Mutabilis: Grant/Research Support|Nabriva Therapeutics AG: Grant/Research Support|Paratek Pharmaceuticals, Inc.: Grant/Research Support|Qpex Biopharma: Grant/Research Support|Sfunga Therapeutics: Grant/Research Support|Spero Therapeutics: Grant/Research Support|Suzhou Sinovent Pharmaceuticals Co.: Grant/Research Support|Theravance: Grant/Research Support|tranScrip Partners: Grant/Research Support|University of Wisconsin: Grant/Research Support|Utility Therapeutics: Grant/Research Support|ValanBio Therapeutics Inc.: Grant/Research Support|VenatoRx: Grant/Research Support Karine Litherland, Ph.D., Basilea Pharmaceutica International Ltd, Allschwil, Switzerland: Full time employee|Basilea Pharmaceutica International Ltd, Allschwil, Switzerland: Stocks/Bonds