Dose-response Curves Research Articles

Indicators of Response to the Wee1 Inhibitor Adavosertib in Acute Myeloid Leukemia

Wee1 is a nuclear kinase that regulates cell cycle progression by inhibiting Cdk1, which is essential for G2 to M phase transition. Inhibition of Wee1 leads to high Cdk1 activity, causing cells to bypass proper DNA repair at the G2/M checkpoint, resulting in mitotic catastrophe and cell death, plus increased Cdk2 activity, causing abnormal DNA replication and DNA double-stranded breaks. Inhibitors of Wee1 are being investigated for the treatment of a variety of cancers including AML. In this study, we aimed to identify biomarkers for Wee1 inhibitor (Wee1i) response in AML using ex vivo drug sensitivity data, bulk RNA sequence, exome sequence, and associated clinical data for a large set of primary AML samples tested against the Wee1i adavosertib. Mononuclear cells from bone marrow aspirates (n=136) or peripheral blood samples (n=11) were collected from AML patients using approved protocols and after informed consent. 101 samples were collected at the time of diagnosis and 35 samples were from relapsed/refractory (R/R) patients. For ex vivo drug-sensitivity testing, freshly isolated cells were tested against a library of up to 634 oncology drugs (215 approved drugs) including adavosertib. The drugs were pre-plated in 5 concentrations in a 10,000-fold concentration range and incubated with cells for 72h. Cell viability was assessed using the CellTiter Glo (CTG) assay, and a drug sensitivity score (DSS) calculated based on the area under the dose response curve. Somatic alterations of the drug tested samples were identified via exome sequencing of DNA from the AML samples and matched skin biopsy controls. Gene expression data were obtained from bulk RNA sequence data of the AML samples. Out of 136 samples analyzed, the median DSS for adavosertib was 8.45. We found positive and significant correlations between sensitivity to adavosertib and two factors: 1) malignant cell percentage (R= 0.31, p-value= 0.0097), and 2) final relative viability percentage (R=0.58, p-value= 4.3e-13). Correlation analysis between the DSS of adavosertib and 633 other drugs revealed several strong positive and significant correlations. These included drugs with specific mechanisms of actions including HSP90 inhibitors (n=5), JAK2 inhibitors (n=4), topoisomerase II inhibitors (n=4), CDK4/6 inhibitors (n=3), mitotic inhibitors, vinca alkaloid microtubule depolymerizers (n=3), and PLK1 inhibitors (n=3). Since adavosertib also targets PLK1 in addition to Wee1, the correlation with PLK1 inhibitor activity confirms the similar target profiles. Based on the specific adavosertib DSS thresholds, the samples were separated in two groups. Samples sensitive to adavosertib (n=35) had DSS between 21.3 (max) and 11.9 (3rd quartile). Samples resistant to adavosertib (n=34) had DSS between 0.0 (min) and 5.65 (1st quartile). Differential gene expression analysis between the sensitive vs resistant samples showed that multiple isoforms of the PCDH and HOXB genes were upregulated in adavosertib sensitive samples. The GLI2, PI15, and SKIDA1 genes were also upregulated in the sensitive samples (Figure A). In contrast, genes such DNTT, PF4V1, TRH, CD1B, S100A16, and CD34 were overexpressed in resistant samples (Figure B). Gene set enrichment analysis revealed positive enrichment of pathways related to MYC targets v2 in the sensitive samples. On the other hand, hallmark pathways associated with cell cycle, apoptosis, RAS signaling, and p53 showed significant negative enrichment scores in the sensitive samples. No statistically significant associations were observed between drug response and frequently occuring somatic mutations in our dataset. However, genetic abberations related MLLT4-MLL/MLL-fusions were only observed in sensitive samples. Collectively, our current findings show that there may be specific gene expression signatures associated with Wee1i sensitivity such high expression of PCDH and HOXB. Also, the positive enrichment of the MYC target v2 pathway in adavosertib sensitive samples suggests that highly proliferating tumor cells might be more sensitive to Wee1i. In addition, high CD34 expression in resistant samples suggests that immature stem and progenitor cells may be less susceptible to Wee1i. In summary, targeting of Wee1 in AML - as studied in NCT05682170 azenosertib study - may be effective for a molecularly defined subset of patients.

Drug Response Profiling Informs Personalized Bridging to Cell Therapy for Patients with Relapsed/Refractory Acute Lymphoblastic Leukemia

Introduction Treatment of relapsed/refractory acute lymphoblastic leukemia (r/r ALL) remains challenging in spite of recent progress with immunotherapy and targeted small-molecule combinations. Growing awareness of the molecular complexity of the disease with subtype-specific cancer dependencies positions functional drug testing as a promising precision tool to improve the prediction of clinical drug efficacy and adapt treatment. We leverage automated, image-based Drug Response Profiling (DRP) and machine learning to deliver individualized therapeutic options in real time for ALL patients with urgent medical needs. Here we report on the DRP experience of 307 primary samples from 278 patients with BCP-ALL, T-ALL/T-LBL and mixed phenotypic acute leukemia (MPAL) registered from 2016 to 2022 in a non-interventional setting. Methods We have generated a reference set of 141 diagnostic samples from patients with high-risk (75%) or standard/medium-risk (25%) disease as defined by the clinical trial AIEOP-BFM ALL 2017, alongside 122 samples with first and 44 with more advanced relapse. We assayed cell viability at single-cell level from viably frozen bone marrow (BM) samples, co-cultured with hTERT-immortalized mesenchymal stromal cells (MSC) and following a 72h drug incubation with a library of up to 110 compounds. We quantified cell counts at five concentrations in duplicates by segmenting and classifying MSC, live and dead leukemia cells using a modular image processing pipeline deployed on cloud infrastructure. We longitudinally confirmed assay consistency with PDX models and monitored integrity of the stromal layer on a per-sample basis. In a subset of cases (15-20%), relevant stromal detachment or low leukemia survival led to samples not meeting all quality control standards. Drugs were ranked by the area under the dose response curve (AUC) and referenced to all previous samples. This patient-specific ex vivo drug fingerprint was reported to the treating physician with a typical turn-around time of less than two weeks. Results Dexamethasone sensitivities measured ex vivo in BM samples from pediatric front-line patients, who were treated on a one-week cytoreductive steroid pre-phase followed by a four-drug induction regimen, correlated significantly with clinical responses at d8 (PGR vs. PPR, p<0.001), d15 (flow MRD below 10%, p<0.001) and d33 (PCR MRD below E-2, p<0.001). By integrating dose-response profiles from all patients, we observed differential drug activities across cytogenetic groups and identified exceptional responders to Venetoclax in very high-risk subtypes including TCF3::HLF (5 patients, AUC percent rank: 0.03 - 0.19) as well as refractory T-ALL. In 77 r/r T-ALL patients from 12 different countries, DRP supported selection of a personalized bridging regimen prior to consolidation with cellular therapy. Top-scoring drug sensitivities (10th percentile of AUC) were found for a variety of agents including Imatinib, Dasatinib, Venetoclax, Bortezomib, Selinexor and different nucleoside analogs. Eleven T-ALL and 5 BCP-ALL patients received a DRP-guided bridging element involving compounds with high ex vivo drug efficacy and complemented by chemotherapy (Table 1). The other 66 r/r patients were treated according to physician's choice incorporating information on drug resistance detected by DRP in real time. Bridging strategies implementing DRP-guided therapy blocks stabilized or reduced disease burden in 15/16 patients allowing HSCT or infusion of CAR-T cells in 12 patients with 8 patients still being in remission at last follow-up. Conclusion We describe an array of pharmacological sensitivity and resistance patterns across a heterogeneous, high-risk cohort of BCP- and T-ALL patients. We identified individual susceptibilities to targeted agents of different classes including BH3-mimetics, proteasome, tyrosine kinase and exportin inhibitors as well as standard-of-care chemotherapeutics. Prospective recording of drug response profiles offers opportunities to systematically correlate ex vivo and clinical responses and inform personalized therapies. These data strongly advocate inclusion of DRP into randomized, early and late phase clinical trials for high-risk pediatric ALL, comprehensively integrating molecular and functional precision information to improve patient stratification and long-term outcome after HSCT and CAR-T.

Cyclosporine and Voclosporin Resistant Immune Effector Cells to Improve Outcomes after Stem Cell Transplantation

Introduction: Relapse of a patient's underlying malignancy, and complications related to viral reactivation are prominent causes of morbidity and mortality after allogeneic hematopoietic cell transplantation (HCT). Immune effector cells (IEC) are increasingly deployed to prevent and treat these complications. These include anti-CD19 CAR-T-cells used for post-HCT B-ALL relapse, as well as viral specific T cells (VSTs) targeting EBV, CMV, and others. However, the efficacy of these IECs early after HCT may be compromised by the inhibitory effects of immunosuppressive agents used to prevent graft-versus host-disease (GVHD). A commonly employed immunosuppressant used for GVHD prophylaxis is the calcineurin inhibitor (CNI) cyclosporine (CsA). A structural analog, voclosporin (VCS), is now FDA approved for lupus nephritis and in trials to prevent rejection after solid organ transplantation. Given its favorable pharmacodynamic and toxicity profile versus CsA and tacrolimus, VCS is poised to become an important option for HCT patients. Both CsA and VCS interact with calcineurin only in the presence of their immunophilin binding partner - cyclophilin A ( PPIA). Tacrolimus and sirolimus interact with a distinct immunophilin, FKBP12. This creates the opportunity to delete a single immunophilin (eg. PPIA) to create IECs resistant to one class of CNI (CsA/VCS) but sensitive to other immunosuppressive agents, an important safety feature. Here we developed CsA/VCS-resistant IECs by CRISPR/Cas9 knockout (KO) of PPIA. These cells exhibited superior proliferation and preserved effector function in the presence of CsA and VCS. Methods: We developed IECs that are simultaneously resistant to both CsA and VCS via CRISPR-mediated KO of PPIA. Proliferation, cytotoxicity, and cytokine production of PPIA KO conventional T cells (Tcon), CMV (pp65)-specific VSTs, and CD19 CAR-T cells were tested against mock-edited (MOCK) cells in functional in vitro assays with escalating doses of CsA and VCS. Results: PPIA KO Tcon demonstrated high editing efficiency (88-92%), high viability, and robust expansion in culture. Indeed, PPIA KO efficiency was maintained over several weeks in culture, suggestive of no survival disadvantage of cells bearing this deletion. Importantly, when PPIA KO cells were used as Responders in a mixed lymphocyte reaction in the presence of CsA, the edited cells demonstrated significant resistance to drug-mediated suppression of CD4+ and CD8+ T cell proliferation across the dose-response curve (p < 0.01; Fig. 1). Similar results were observed with VCS, with PPIA KO cells significantly resistant to VCS-mediated suppression across otherwise highly suppressive doses (p<0.0001; Fig. 1). Having demonstrated that edited Tcon were resistant to CsA and VCS, we next sought to make PPIA KO IECs. CRISPR-edited VSTs and CD19 CAR-T cells showed similar editing efficiencies to Tcon. Consistent with prior results, CMV-specific PPIA KO responder VSTs were significantly resistant to suppression of proliferation by CsA vs MOCK cells at all doses tested (p=0.0004, not shown). Proliferation in the presence of drug was then assessed in KO CAR-T cells using the CD19+ NALM6 line as stimulators. Division of PPIA KO responder CD19 CAR-T cells was almost completely unaffected by CsA and VCS across escalating doses ( Fig. 2). For example, at an effector-to-target (E:T) ratio of 1:0.1 and a CsA dose of 50 ng/mL, there was 40.6 ± 2.3 % suppression of division in MOCK CD8+ CAR-T cells vs. -0.2 ± 0.4 % in PPIA KO CD8+ CAR-T cells (p<0.0001). Similar findings were observed with CD4+ responder CAR-T cells (p<0.0001, not shown). PPIA KO CAR-T cells also demonstrated intact cytotoxic function with no significant differences in % killing between the MOCK and PPIA KO groups at 3 different E:T ratios. For example, at a 1:1 E:T ratio, there was 59 ± 15% killing vs. 55 ± 12% killing in MOCK vs PPIA KO groups, respectively (not shown). Conclusion: KO of PPIA is an effective strategy for engineering resistance to CsA and VCS incellular therapies. Our results demonstrate that PPIA KO conventional T cells, VSTs, and CAR-T cells were markedly resistant to the otherwise suppressive effects on T cell proliferation that are hallmarks of both CsA and VCS. Engineering CNI-resistance into IECs will enhance the ability to deploy these cells early after transplant to prevent or treat both malignant relapse and viral reactivation.

Lysosomal Membrane Permeabilization Sensitizes Ctss-Hyperactive Tumors to BCL2-Targeting Therapies

Hyperactivity of the cysteine protease cathepsin S (CTSS) -either through Y132 mutations or amplification/overexpression- is a recurrent alteration in follicular lymphoma (FL) and promotes tumor growth by inducing a supportive immune microenvironment (Bararia et al, 2020). Of note, patients with CTSS-hyperactive FL had better outcomes with standard therapies, suggesting that CTSS-hyperactivity can sensitize tumors to treatment. CTSS hyperactivity has also been reported in other B cell lymphomas (BCLs) (Dheilly et al, 2020) and solid cancers (Olson & Joyce, 2015). CTSS is mainly localized intralysosomally but can be released into the cytosol by lysosomal membrane permeabilization (LMP). Low level LMP can occur spontaneously (e.g., during cell division) and can be enhanced by treatment. Unlike other cathepsins, cytosolically released CTSS maintains its enzymatic activity at non-acidic pH. Thus, we aimed to (i) identify the determinants of the cytosolic CTSS activity, (ii) determine its impact on the regulation of apoptosis, and (iii) study LMP as a therapeutic approach for CTSS-hyperactive tumors. First, we accrued biochemical, functional, and clinical data supporting the role of cystatin B (CSTB) as a clinically relevant endogenous CTSS inhibitor in BCLs. Through unbiased and complementary proteomics (BioID2 labelling and co-IP followed by LC-MS/MS) we identified CSTB as a direct CTSS-interacting protein (8-fold enriched in the BCL cell line Karpas422 engineered to express CTSS wild type (WT) or Y132D vs CTSS knock-out (KO), P=0.0002). Single-cell RNA-Seq of primary FL (N=10) showed significantly higher CSTB expression in FL cells compared to normal B cells ( P=0.004). Moreover, shRNA mediated knock-down (k/d) of CSTB increased the overall cathepsin activity in BCL cell lines (N=8) by 1.5 to 5.5-fold, most notably in CTSS-hyperactive cells ( Fig A, top). We next employed LMP-inducing tool compounds (LLOMe) and clinically used drugs or analogs (desipramine, hexamethylene amiloride) to release cathepsins into the cytosol. CTSS-hyperactive Karpas422 were significantly more sensitive to LMP-inducing treatments compared to native cells (1.5 to 10-fold reduction of IC50). Importantly, CTSS hyperactivity and CSTB k/d increased LMP-mediated cell killing ( Fig A, bottom). Thus, the cytosolic CTSS/CSTB interaction determines the net cytosolic cathepsin activity and sensitivity of cells to undergo LMP-induced cell death. Next, we hypothesized that LMP-induced cytosolic CTSS hyperactivity could prime BCLs towards apoptosis. We used BH3 profiling to functionally quantify the dependencies and interactions of BCL2 family members in BCLs with and without CTSS hyperactivity. In Karpas422 cells expressing CTSS Y132D, LMP increased overall apoptotic priming and dependencies on the anti-apoptotic proteins MCL-1 (delta priming >30 % at 10 µM, P=0.04), BCL-xL (>45 % at 10 µM, P=0.0006) and BCL2 (> 45 % at 0.5 and 1 µM, P=0.0001). We hypothesized that BCL2 family members are proteolytically cleaved by cytosolic CTSS. Indeed, e.g., BCL2 protein levels were 2.5 to 3.5-fold lower in LLOMe-treated Karpas422 cells with CTSS-hyperactivity compared to CTSS KO, and CSTB k/d further decreased BCL2 levels. To validate CTSS-mediated cleavage of BCL2, we purified FLAG-tagged BCL2 and CTSS WT and Y132D. CTSS WT efficiently cleaved BCL2 in vitro <1 hour at the top ranked predicted cleavage site and the reaction rate increased 1.3-fold for CTSS Y132D. Finally, we hypothesized that LMP sensitizes cells to BCL2-targeting therapies ( Fig B). The combination of LLOMe-induced LMP and the BCL2 inhibitor venetoclax (VEN) showed increased cytotoxicity in CTSS-hyperactive Karpas422 cells compared to monotherapy and CSTB k/d enhanced this phenotype ( Fig A, bottom). We assessed cathepsin activities and generated dose-response curves for VEN with and without LLOMe-induced LMP in 15 primary CLL samples. Thereof, 12 samples had intermediate or high cathepsin activities and LLOMe-induced LMP increased their sensitivity to VEN, including a VEN-resistant sample in which the IC50 decreased to <3 nM. In summary, we show that CSTB is a functionally relevant inhibitor that determines the net activity of LMP-released cytosolic CTSS. Furthermore, LMP-inducing therapies may be a promising approach to sensitize CTSS-hyperactive tumors towards apoptosis by proteolytic cleavage of BCL2 family members.

Associations of plasma metals/metalloids with arrhythmia among occupational population: a prospective study

To investigate the association between levels of twenty-three plasma metals/metalloids and the risk of arrhythmia among occupational population. In 2017, a total of 765 workers aged 18 and above were recruited from a non-ferrous metal factory. The general demographic characteristics were obtained by using questionnaire. Plasma metal/metalloid levels were determined by inductively coupled plasma mass spectrometry(ICP-MS). Participants were followed up in 2018, 2019 and 2020 respectively. After the elements that may affect the incidence of arrhythmia were screened out by least absolute shrinkage and selection operator(LASSO) regression, Cox regression model was used to analyze the relationship between levels of selected elements and risk of arrhythmia occurrence, Quantile g-computation model was used to analyze the effect of element mixture exposure on arrhythmia, and the dose-response curve was estimated by using restricted cubic spline(RCS) function. Of all the research subjects, 386(50.5%) were ≤45 years old; 401(52.4%) had 20 years or more of work experience; 712(93.1%) subjects were male workers. The incidence of arrhythmia was 17.6%. After adjusting for age, seniority, gender, body mass index(BMI), marital status, education level, smoking, drinking, drinking tea, regular exercise, chronic diseases(hypertension, hyperlipidemia), sleep quality and psychological stress, chromium, molybdenum and antimony increased the risk of arrhythmia with HR(95%CI) values of 1.22(1.11-1.34), 1.51(1.20-1.90) and 2.38(1.03-5.49), respectively, while barium reduced the risk of arrhythmia with HR(95%CI) value of 0.98(0.95-1.00). Chromium, molybdenum and antimony are the risk factors while barium is the protective factor for arrhythmia.

Association of serum copper (Cu) with cardiovascular mortality and all-cause mortality in a general population: a prospective cohort study

BackgroundCopper (Cu) homeostasis and Cu-induced cell death are gaining recognition as crucial processes in the pathogenesis of cardiovascular disease (CVD). Circulating Cu associated with CVD and mortality is yet to be fully elucidated.ObjectiveThis national prospective cohort study is to estimate relationship between serum Cu and the risk of CVD and all-cause mortality.MethodsThis study included participants from the National Health and Nutrition Examination Survey 2011–2016. Weighted Cox proportional hazards regression analysis and exposure-response curves were applied.ResultsThis included 5,412 adults, representing 76,479,702 individuals. During a mean of 5.85 years of follow-up (31,653 person-years), 96 CVD and 356 all-cause mortality events occurred. Age and sex-adjusted survival curves showed that individuals with higher levels of serum Cu experienced increased CVD and all-cause death rates (tertiles, p < 0.05). Compared with the participant in tertile 1 of serum Cu (< 16.31 mol/L), those in tertile 3 (≥ 19.84 mol/L) were significantly associated with CVD mortality (HR: 7.06, 95%CI: 1.85,26.96), and all-cause mortality (HR: 2.84, 95% CI: 1.66,4.87). The dose-response curve indicated a linear relationship between serum Cu and CVD mortality (p -nonlinear = 0.48) and all-cause (p -nonlinear = 0.62). A meta-analysis included additional three prospective cohorts with 13,189 patients confirmed the association between higher serum Cu and CVD (HR: 2.08, 95% CI: 1.63,2.65) and all-cause mortality (HR: 1.89, 95%CI: 1.58,2.25).ConclusionThe present study suggests excessive serum Cu concentrations are associated with the risk of CVD and all-cause mortality in American adults. Our findings and the causal relationships require further investigation.Graphical

Preliminary research on the dose response of polymer gel dosimeter with carbon beam irradiation

This paper is to investigate dose response of polymer gel dosimeter after carbon irradiation. We employed polymer gel dosimeter, MAGAT-f to measure the dose response curve, single spot shape and Bragg peak range of carbon beam. Irradiation energy was 80.55 MeV/u. In order to characterize the dose response, R2-dose curve and absorbance-dose curve were built up. An approximate linearity is found for R2-dose relationship. Saturation is found for the absorbance of the gel after 10 Gy. The results illustrate the feasibility of utilizing polymer gel dosimeter for Bragg peak and spot shape measurement of carbon ion beam.

Dose-Response Relationship Between Radiation Therapy and Loss of Lung Perfusion Comparing Positron Emission Tomography and Dual-Energy Computed Tomography in Non-Small Cell Lung Cancer

Background: Radiotherapy treatment for non-small cell lung cancer (NSCLC) may result in radiation damage to the perfused lung. The loss in perfusion may be measured from positron tomography emission (PET) perfusion imaging, however, this modality may not be widely available. Dual-energy computed tomography (DECT) with contrast may be an alternative to PET/CT. The purpose of this work is to investigate the equivalence of dose-response curves (DRC) determined from PET and DECT in NSCLC. Methods: PET and DECT datasets from the prospective clinical trial BLINDED FOR REVIEW (BLINDED FOR REVIEW) were included in this pre-planned trial analysis. Patients underwent 68Ga-macroaggregated albumin (68Ga-MAA) PET/CT examination and DECT with contrast on the same day at baseline/3-months/12-months post-treatment. The perfused lung was defined from a threshold based on the maximum standardized uptake value (%SUVmax)/iodine concentration (%IoMax) in PET/DECT. The equivalence between PET and DECT DRC was established by comparing (1) the average of the normalized overlap of the two DRCs (aNO), ranging from 0 (no overlap) to 1 (perfect overlap), and (2) the slope of a linear model applied to DRCs. Results: Out of the 19 patients enrolled in the clinical trial, 14/10 patients had a post-treatment imaging session at median = 4.5 months/13.5 months, respectively. With 30%SUVmax/35%IoMax, aNO was maximized and the difference between PET and DECT slope of the linear model was minimized at each time point (slope = 0.76%/Gy / 0.75%/Gy at 3 months and 0.86%/Gy / 0.87%/Gy at 12 months determined from PET / DECT). Conclusions: Dose-response relationship determined from DECT was comparable to PET at 3- and 12-months post-treatment in NSCLC patients.

Additive Effects of Helix aspersa Mucus Fractions with Chemotherapy Drugs in Breast Cancer: Exploring Anti-proliferative Potential

This study investigated the anti-proliferative effects of biologically active substances (BAS) derived from the mucus of the garden snail Helix aspersa on breast cancer cell lines (MCF-7 and MDA-MB-231) and a non-cancerous cell line (MCF-10A). Two mucus fractions with different molecular weights (Mw) were examined, and their interactions with the chemotherapy drugs cisplatin and tamoxifen were evaluated.&#x0D; The results showed that the mucus fractions exhibited a slight anti-proliferative effect on breast cancer cells, with the fraction having Mw above 50 kDa demonstrating stronger activity compared to the fraction with Mw above 20 kDa. Furthermore, the combination of the mucus fractions with cisplatin or tamoxifen resulted in significant reductions in the half effective dose, indicating possible synergistic effects which were more pronounced with the fraction having Mw above 20 kDa. Dose-response curves and microscopic images supported the presence of such effects, revealing lower IC50 values and significant cell death when the cancer cells were treated with the combination of the mucus fractions and chemotherapy drugs. However, the analysis using the (highest single agent) HSA model indicated mostly additive interactions between the mucus fractions and chemotherapy drugs.&#x0D; These findings suggest the potential of biologically active substances derived from Helix aspersa mucus in enhancing the anti-proliferative effects of chemotherapy drugs in breast cancer treatment. The identification of additive interactions provides insights into the development of combination therapies with reduced drug dosage. Overall, this study contributes to the understanding of the anti-proliferative effects of Helix aspersa mucus fractions and their interactions with chemotherapy drugs, highlighting their potential as novel therapeutic approaches in breast cancer treatment.

Sex difference in the non-linear relationship between ethylene oxide exposure and depressive symptoms: A cross-sectional study

BackgroundEthylene oxide (EO) has been recognized as an animal carcinogen and environmental EO exposure was linked to several diseases. However, the association of EO exposure with depression prevalence is still not clear. MethodsWe included 6016 participants with complete data on HbEO concentrations, depression diagnosis, and necessary covariates using the 2013–2020 National Health and Nutrition Examination Survey. Weighted multivariable logistic model was applied to examine the association of HbEO concentrations with depression risk. Weighted restricted cubic spline model was applied to draw the dose-response curve. ResultsIn the total population, individuals in the second, third, and fourth quartile of HbEO respectively had an adjusted OR of 0.99 (95%CI: 0.60, 1.63), 1.13 (95%CI: 0.73, 1.75), and 2.87 (95%CI: 1.86, 4.45) (Ptrend < 0.001) for depression with a significant “J” shaped non-linear dose-response relationship (Pnon-linear < 0.001). Females, drinkers, and smokers were susceptible to the depressive effect of EO. Doubling the HbEO concentrations was respectively associated with a 1.50-fold (95%CI: 1.25, 1.79), 1.29-fold (1.15, 1.44), and 1.17-fold (1.04, 1.33) increased risk of depression for females, drinkers, and smokers. LimitationsCross-sectional study design and self-reported depressive symptoms. ConclusionsEnvironmental EO exposure was associated with increased depression risk, especially among females, drinkers, and smokers. Further prospective studies are required to affirm these findings.

Targeted randomization dose optimization trials enable fractional dosing of scarce drugs.

Administering drug at a dose lower than that used in pivotal clinical trials, known as fractional dosing, can stretch scarce resources. Implementing fractional dosing with confidence requires understanding a drug's dose-response relationship. Clinical trials aimed at describing dose-response in scarce, efficacious drugs risk underdosing, leading dose-finding trials to not be pursued despite their obvious potential benefit. We developed a new set of response-adaptive randomized dose-finding trials and demonstrate, in a series of simulated trials across diverse dose-response curves, these designs' efficiency in identifying the minimum dose that achieves satisfactory efficacy. Compared to conventional designs, these trials have higher probabilities of identifying lower doses while reducing the risks of both population- and subject-level underdosing. We strongly recommend that, upon demonstration of a drug's efficacy, pandemic drug development swiftly proceeds with response-adaptive dose-finding trials. This unified strategy ensures that scarce effective drugs produce maximum social benefits.

A Method for Ensuring a Consistent Dose–Response Relationship Between An Entire Population and One Region in Multiregional Dose–Response Studies Using MCP-Mod

Multiregional clinical studies are frequently conducted worldwide to accelerate drug approval. Conducting multiregional dose–response studies is useful to determine the effects of important ethnic factors on a drug during the early stages of development. Sample size allocation for regions should be based on a scientific rationale. Recently, Yamaguchi and Sugitani proposed a method for sample size allocation for multiregional dose–response studies using MCP-Mod. This method ensures that a consistent dose–response relationship in the contrast test statistic and the maximum dose–response curve difference between the entire population and one region are observed with sufficient probability. However, this method is not easy to use because of the inability to compute the assurance probability of consistency. In this study, we propose an analytical solution by defining the probability formulas for the existing method. The difficulty lies in formulating the probability of maximum curve differences between two nested populations: the entire population and one specific region. We addressed this difficulty by approximating the curve of the entire population by combination of the independent regional curves. The proposed formula enables the computation of the probability based on a multivariate normal distribution. The accuracy of the proposed formulas was confirmed through a simulation study.

Relationship between weight-adjusted waist circumference index and prevalence of gallstones in U.S. adults: a study based on the NHANES 2017-2020.

We aimed to assess the association between weight-adjusted waist circumference index (WWI) and gallstone prevalence in US adults. We selected individuals from the National Health and Nutrition Examination Survey (NHANES) database from 2017 to 2020 and used logistic regression analyses, subgroup analyses, and dose-response curves to assess the association between WWI and gallbladder stone prevalence and age, sex, and ethnicity. A total of 7971 participants aged ≥20 years were enrolled in our study; 828 patients had a self-reported history of gallstones. After correcting for confounders, for each unit of WWI after Ln conversion, the prevalence of gallbladder stones increased by 34% (OR=1.34, 95% CI:1.20, 1.50). Dose-response curves showed a positive correlation between WWI and gallbladder stone prevalence.According to the subgroup analysis, the positive association between TyG index and high-frequency HI was more significant in males(OR=1.34, 95% CI:1.07, 1.69), <40 years old(OR=1.42, 95% CI:1.18, 1.71), white people Americans(OR=1.35, 95% CI:1.08, 1.68) and other races(OR= 1.56, 95% CI:1.13, 2.14). Higher WWI was positively associated with the prevalence of gallbladder stones and was associated with gender, age, and ethnicity. This is noteworthy, although a causal relationship could not be established.

Predictive value of nutritional indices for left atrial thrombus in patients with valvular atrial fibrillation

BackgroundThe prognostic nutritional index (PNI) and geriatric nutritional risk index (GNRI) are well known indicators for adverse outcomes in various diseases, but there is no evidence on their association with the risk of left atrial thrombus (LAT) in patients with valvular atrial fibrillation (VAF).MethodsA comparative cross-sectional analytical study was conducted on 433 VAF patients. Demographics, clinical characteristics and echocardiographic data were collected and analyzed. Patients were grouped by the presence of LAT detected by transesophageal echocardiography.ResultsLAT were identified in 142 patients (32.79%). The restricted cubic splines showed an L-shaped relationship between PNI and LAT. The dose-response curve flattened out near the horizontal line with OR = 1 at the level of 49.63, indicating the risk of LAT did not decrease if PNI was greater than 49.63. GNRI was negative with the risk of LAT and tended to be protective when greater than 106.78. The best cut-off values of PNI and GNRI calculated by receiver operating characteristics curve to predict LAT were 46.4 (area under these curve [AUC]: 0.600, 95% confidence interval [CI]:0.541–0.658, P = 0.001) and 105.7 (AUC: 0.629, 95% CI:0.574–0.684, P<0.001), respectively. Multivariable logistic regression analysis showed that PNI ≤ 46.4 (odds ratio: 2.457, 95% CI:1.333–4.526, P = 0.004) and GNRI ≤ 105.7 (odds ratio: 2.113, 95% CI:1.076–4.149, P = 0.030) were independent predictors of LAT, respectively.ConclusionsLower nutritional indices (GNRI and PNI) were associated with increased risk for LAT in patients with VAF.

Association between polyunsaturated fatty acid intake and the prevalence of erectile dysfunction: A cross-sectional analysis of the NHANES 2001–2004

BackgroundPolyunsaturated fatty acids (PUFAs) have demonstrated significant therapeutic potential across a wide range of disease. The aim of this study was to investigate the potential impact of PUFA intake on the prevalence of erectile dysfunction (ED).MethodsThe study included a total of 3730 participants from the National Health and Nutrition Examination Survey (NHANES) 2001–2004. Univariate analysis, multivariate regression analysis, subgroup analysis and machine learning were utilized to explore the relationship of variables to ED. Dose response curves were constructed to observe the linear or nonlinear relationship between PUFA intake and the prevalence of ED. Propensity score matching (PSM) was used for sensitivity analysis. Finally, the potential mechanistic link between PUFA intake and ED was explored.ResultsThrough univariate and multivariate regression analysis results before and after PSM and XGBoost algorithm model results, arachidonic acid (AA) was chosen as the main research object. The consumption of AA was found to be associated with a decreased prevalence of ED under the fully adjusted model [OR = 0.33 (0.20, 0.56), P < 0.001]. The interaction between AA and education was found in the subgroup analysis. Dose-response curves indicated a linear negative correlation between AA intake and the prevalence of ED. The above results were confirmed in the data analysis after 1:1 PSM. In addition, AA intake was associated with a decrease in inflammatory biomarkers and homocysteine.ConclusionsThe results suggest that AA intake is negatively correlated with the prevalence of ED. Further, anti-inflammatory and anti-endothelial damage may play a role in this.

Treatment Outcomes With Licensed and Unlicensed Stimulant Doses for Adults With Attention-Deficit/Hyperactivity Disorder

Stimulants (methylphenidate and amphetamines) are often prescribed at unlicensed doses for adults with attention-deficit/hyperactivity disorder (ADHD). Whether dose escalation beyond US Food and Drug Administration recommendations is associated with positive risk benefits is unclear. To investigate the impact, based on averages, of stimulant doses on treatment outcomes in adults with ADHD and to determine, based on averages, whether unlicensed doses are associated with positive risk benefits compared with licensed doses. Twelve databases, including published (PubMed, Cochrane Library, Embase, Web of Sciences) and unpublished (ClinicalTrials.gov) literature, up to February 22, 2023, without language restrictions. Two researchers independently screened records to identify double-blinded randomized clinical trials of stimulants against placebo in adults (18 years and older) with ADHD. Aggregate data were extracted and synthesized in random-effects dose-response meta-analyses and network meta-analyses. Change in ADHD symptoms and discontinuations due to adverse events. A total of 47 randomized clinical trials (7714 participants; mean age, 35 (SD, 11) years; 4204 male [56%]) were included. For methylphenidate, dose-response curves indicated additional reductions of symptoms with increments in doses, but the gains were progressively smaller and accompanied by continued additional risk of adverse events dropouts. Network meta-analyses showed that unlicensed doses were associated with greater reductions of symptoms compared with licensed doses (standardized mean difference [SMD], -0.23; 95% CI, -0.44 to -0.02; very low certainty of evidence), but the additional gain was small and accompanied by increased risk of adverse event dropouts (odds ratio, 2.02; 95% CI, 1.19-3.43; moderate certainty of evidence). For amphetamines, the dose-response curve approached a plateau and increments in doses did not indicate additional reductions of symptoms, but there were continued increments in the risk of adverse event dropouts. Network meta-analysis did not identify differences between unlicensed and licensed doses for reductions of symptoms (SMD, -0.08; 95% CI, -0.24 to 0.08; very low certainty of evidence). Based on group averages, unlicensed doses of stimulants may not have positive risk benefits compared with licensed doses for adults with ADHD. In general, practitioners should consider unlicensed doses cautiously. Practitioners may trial unlicensed doses if needed and tolerated but should be aware that there may not be large gains in the response to the medication with those further increments in dose. However, the findings are averages and will not generalize to every patient.

The use of in silico molecular modelling to screen potential estrogen mimics as part of medicines and agrochemicals development and product license applications.

Estrogen mimics are a diverse group of synthetic and naturally occurring compounds that can interact with estrogen receptors (ERs) in animals, including humans. These interactions rely on key structural features of 17b-estradiol (E2) and if these molecular features are in a similar spatial arrangement on other compounds, they are likely to elicit an agonist (i.e., they are E2 mimics) or antagonist response. The structural diversity of some compounds vis-à-vis analogies with E2 makes it difficult to reliably predict E2 mimicry on simple structural grounds alone. We propose a new approach methodology: in silico molecular modelling augmented by an in vitro transactivation reporter gene assay to predict E2 mimicry and thus further reduce regulatory reliance on animal studies. Transactivation reporter gene assay dose response curves and in silico molecular modelling were used to obtain EC50-values and docking parameters (DockScores), respectively of thirty E2 mimics to assess the reliability of in silico receptor interaction parameters to predict E2 mimicry. A linear relationship (R2 = 0.75) was found between DockScores and EC50s, suggesting molecular modelling is a good tool for predicting E2 mimicry in a regulatory setting.

Microfluidic Sensor Based on Cell-Imprinted Polymer-Coated Microwires for Conductometric Detection of Bacteria in Water.

The rapid, inexpensive, and on-site detection of bacterial contaminants using highly sensitive and specific microfluidic sensors is attracting substantial attention in water quality monitoring applications. Cell-imprinted polymers (CIPs) have emerged as robust, cost-effective, and versatile recognition materials with selective binding sites for capturing whole bacteria. However, electrochemical transduction of the binding event to a measurable signal within a microfluidic device to develop easy-to-use, compact, portable, durable, and affordable sensors remains a challenge. For this paper, we employed CIP-functionalized microwires (CIP-MWs) with an affinity towards E. coli and integrated them into a low-cost microfluidic sensor to measure the conductometric transduction of CIP-bacteria binding events. The sensor comprised two CIP-MWs suspended perpendicularly to a PDMS microchannel. The inter-wire electrical resistance of the microchannel was measured before, during, and after exposure of CIP-MWs to bacteria. A decline in the inter-wire resistance of the sensor after 30 min of incubation with bacteria was detected. Resistance change normalization and the subsequent analysis of the sensor's dose-response curve between 0 to 109 CFU/mL bacteria revealed the limits of detection and quantification of 2.1 × 105 CFU/mL and 7.3 × 105 CFU/mL, respectively. The dynamic range of the sensor was 104 to 107 CFU/mL where the bacteria counts were statistically distinguishable from each other. A linear fit in this range resulted in a sensitivity of 7.35 μS per CFU/mL. Experiments using competing Sarcina or Listeria cells showed specificity of the sensor towards the imprinted E. coli cells. The reported CIP-MW-based conductometric microfluidic sensor can provide a cost-effective, durable, portable, and real-time solution for the detection of pathogens in water.

Association of mean arterial pressure with non-alcoholic fatty liver disease: results from the NAGALA study.

Several recent reports have suggested the use of mean arterial blood pressure (MAP) to assess/predict the risk of developing atherosclerosis, chronic kidney disease, diabetes, metabolic syndrome, and poor prognosis in a variety of cardiovascular and cerebrovascular diseases. The current study aimed to investigate the association of MAP with non-alcoholic fatty liver disease (NAFLD) and to explore the differences in this association across populations. This study used data from the NAGALA study from 1994 to 2016. MAP was calculated as 1/3 systolic blood pressure (SBP) + 2/3 diastolic blood pressure (DBP). Restricted cubic spline (RCS) and logistic regression models were used to examine the correlation of MAP with NAFLD. The study population was 14,251 general people undergoing health screening, with a median (interquartile range) age of 42 (36-50) years; among them, 48% were women, and 2,507 (17.59%) were diagnosed with NAFLD. After fully controlling for confounders in the current dataset, MAP was positively and non-linearly associated with NAFLD [(odds ratios (ORs): 1.39, 95% confidence intervals (CIs): 1.15, 1.68); P for non-linearity = 0.024]; the dose-response curve showed that there was a transient saturation effect interval when MAP was between 85 and 95 mmHg, where the risk of NAFLD was neither increased nor decreased. The results of the stratified analysis showed that the risk of NAFLD associated with MAP appeared to be influenced only by age (P-interaction = 0.002), but not by sex, body mass index (BMI), habits of exercise, drinking status, or smoking status (P-interaction > 0.05); further age-stratified RCS analysis showed that the non-linear association between MAP and NAFLD in the young and middle-aged and the middle-aged and elderly populations was consistent with the results of the whole population, whereas, in the elderly population, a U-shaped curve association between MAP and NAFLD was observed, with both low and high MAP increasing the risk of NAFLD. In the general population, MAP was positively and non-linearly associated with NAFLD, and this association only differed significantly by age, but not by sex, BMI, habits of exercise, drinking status, and smoking status.

Fluoride Exposure and Skeletal Fluorosis: a Systematic Review and Dose-response Meta-analysis.

We performed a systematic review and meta-analysis on the relation between fluoride exposure and skeletal fluorosis (SF) using a novel statistical methodology for dose-response modeling. Skeletal fluorosis, a major health issue that is endemic in some regions, affects millions of people worldwide. However, data regarding the dose-response relation between fluoride exposure and SF are limited and outdated. We included twenty-three studies in the meta-analysis. When comparing the highest versus the lowest fluoride category, the summary risk ratio (RR) for SF prevalence was 2.05 (95% CI 1.60; 2.64), with a value of 2.73 (95% CI 1.92; 3.90) for drinking water and 1.40 (95% CI 0.90; 2.17) for urinary fluoride. The RR by the risk of bias (RoB) was 2.37 (95% CI 1.56; 3.58) and 1.78 (95% CI 1.34; 2.36) for moderate and high RoB studies, respectively. The dose-response curve based on a one-stage cubic spline regression model showed an almost linear positive relation between exposure and SF occurrence starting from relatively low concentrations up to 5 mg/L and 2.5 mg/L, respectively, for water and urinary fluoride, with no substantial increase above this threshold. The RR for developing moderate-severe forms increases at 5.00 mg/L and 2.5 mg/L of water and urinary fluoride, respectively. Better-quality studies are needed to confirm these results, but greater attention should be given to water fluoride levels to prevent SF, in addition to the other potential adverse effects of fluoride exposure.