Dose-response Curves Research Articles

Association between the C-reactive protein-triglyceride glucose index and erectile dysfunction in US males: results from NHANES 2001-2004.

The C-reactive protein-triglyceride glucose index (CTI) is emerging as a novel indicator for comprehensively assessing the severity of both inflammation and insulin resistance. However, the association between CTI and erectile dysfunction (ED) remains largely unexplored. Participant data for this study were sourced from NHANES 2001-2004, with exclusion criteria applied to those lacking information on clinical variables. The CTI was defined as 0.412*Ln (CRP) + ln [T.G. (mg/dL) × FPG (mg/dL)/2]. Weighted univariable and multivariable logistic regression models were utilized to examine the correlation between the CTI and ED, assessing the CTI as both a continuous and categorical variable (quartile). Moreover, subgroup analyses were conducted to pinpoint sensitive populations, and interaction analysis was performed to validate the findings. A total of 1502 participants were included in the final analysis, encompassing 302 with ED and 1200 without ED. After adjusting for potential confounders, the CTI was positively associated with ED incidence (OR = 1.56, 95% CI: 1.27-1.90, P = 0.002). The fourth quartile of the CTI significantly increased the incidence of ED (OR = 2.69, 95% CI: 1.07-6.74, P = 0.04), and the lowest quartile of CTI was used as the reference. The dose-response curve revealed a positive linear relationship between the CTI and the incidence of ED. Subgroup analysis confirmed the consistent positive relationship between the CTI and ED. The interaction test indicated no significant impact on this association. Finally, a sensitivity analysis was performed to verify the significant positive correlation between the CTI and severe ED (OR = 1.44, 95% CI: 1.19-1.76, P = 0.004). Our national data indicate that a greater CTI is positively linked to an increased risk of ED in US men, suggesting its potential for use in clinical practice for ED prevention or early intervention. Additional large-scale prospective studies are warranted to substantiate the causative relationship between CTI and ED.

Controlled CMS Data Demonstrates a Cost and Clinical Advantage for Hyperbaric Oxygen for Radiation Cystitis

ABSTRACT Feldmeier J, Kirby J, Gelly H, Robins M, Peters J, Gruhn P, Pal S. Controlled CMS Data Demonstrates a Cost and Clinical Advantage for Hyperbaric Oxygen for Radiation Cystitis. Undersea Hyperb Med. 2024 Second Quarter; 51(2):145-157. Introduction: Increasing cancer survivorship, in part due to new radiation treatments, has created a larger population at risk for delayed complications of treatment. Radiation cystitis continues to occur despite targeted radiation techniques. Materials and Methods: To investigate value-based care applying hyperbaric oxygen (HBO2) to treat delayed radiation cystitis, we reviewed public-access Medicare data from 3,309 patients from Oct 1, 2014, through Dec 31, 2019. Using novel statistical modeling, we compared cost and clinical effectiveness in a hyperbaric oxygen group to a control group receiving conventional therapies. Results: Treatment in the hyperbaric group provided a 36% reduction in urinary bleeding, a 78% reduced frequency of blood transfusion for hematuria, a 31% reduction in endoscopic procedures, and fewer hospitalizations when study patients were compared to control. There was a 53% reduction in mortality and reduced unadjusted Medicare costs of $5,059 per patient within the first year after completion of HBO2 treatment per patient. When at least 40 treatments were provided, cost savings per patient increased to $11,548 for the HBO2 study group compared to the control group. This represents a 37% reduction in Medicare spending for the HBO2-treated group. We also validate a dose-response curve effect with a complete course of 40 or more HBO2 treatments having better clinical outcomes than those treated with fewer treatments. Conclusion: These data support previous studies that demonstrate clinical benefits now with cost- effectiveness when adjunctive HBO2 treatments are added to routine interventions. The methodology provides a comparative group selected without bias. It also provides validation of statistical modeling techniques that may be valuable in future analysis, complementary to more traditional methods.

Spread-out Bragg peak FLASH: quantifying normal tissue toxicity in a murine model.

A favorable effect of ultra-high dose rate (FLASH) radiation on normal tissue-sparing has been indicated in several preclinical studies. In these studies, the adverse effects of radiation damage were reduced without compromising tumor control. Most studies of proton FLASH investigate these effects within the entrance of a proton beam. However, the real advantage of proton therapy lies in the Spread-out Bragg Peak (SOBP), which allows for giving a high dose to a target with a limited dose to healthy tissue at the entrance of the beam. Therefore, a clinically relevant investigation of the FLASH effect would be of healthy tissues within a SOBP. Our study quantified the tissue-sparing effect of FLASH radiation on acute and late toxicity within an SOBP in a murine model. Radiation-induced damage was assessed for acute and late toxicity in the same mice following irradiation with FLASH (Field dose rate of 60 Gy/s) or conventional (CONV, 0.34 Gy/s) dose rates. The right hindleg of unanesthetized female CDF1 mice was irradiated with single-fraction doses between 19.9-49.7 Gy for CONV and 30.4-65.9 Gy for FLASH with 5-8 mice per dose. The leg was placed in the middle of a 5 cm SOBP generated from a mono-energetic beam using a 2D range modulator. Acute skin toxicity quantified by hair loss, moist desquamation and toe separation was monitored daily within 29 days post-treatment. Late toxicity of fibrotic development measured by leg extendibility was monitored biweekly until 30 weeks post-treatment. Comparison of acute skin toxicity following radiation indicated a tissue-sparing effect of FLASH compared to conventional single-fraction radiation with a mean protection ratio of 1.40 (1.35-1.46). Fibrotic development similarly indicated normal tissue sparing with a 1.18 (1.17-1.18) protection ratio. The acute skin toxicity tissue sparing was similar to data from entrance-beam irradiations of Sørensen etal. (4). Full dose-response curves for acute and late toxicity after CONV and FLASH radiation were obtained. Radiation within the SOBP retains the normal-tissue-sparing effect of FLASH with a dose-modifying factor of 40% for acute skin damage and 18% for fibrotic development.

Effect of dietary supplementation of macadamia oil on the growth, immune function, physio-biochemical componentsand thyroid activity of growing rabbits.

The current research aimed to assess the effects of dietary macadamia oil (MO) on carcass traits, growth performance, physio-biochemical components, immune function, thyroid hormonesand inflammation markers of growing rabbits. A total of 96 growing rabbits were randomly distributed into four treatments, with 24 rabbits in each group. The rabbits were fed a basal diet (control group) or a diet supplemented with MO at 0.5 (MO0.5), 1 (MO1.0)and 2 (MO2.0) mL/kg of diet for eight weeks. The daily body weight gain and feed conversion ratio showed a quadratic improvement with increasing levels of MO, and the optimal dose was 1.25 mL/kg of diet. Increasing levels of MO also had a quadratic effect on hepatic and renal functions. Dose-response curves revealed that the optimal doses of MO were 1.50, 1.75and 1.25 mL/kg of diet for total bilirubin, gamma-glutamyl transferase, and creatininerespectively. A quadratic relationship was observed between the increased levels of MO and tumour necrosis factor-α (p = 0.038), interleukin-6 (p = 0.014)and immunoglobulins (p = 0.016 and IgM p = 0.026). Additionally, a linear relationship was observed between the increment in MO levels and both nitric oxide (p = 0.040) and interleukin-4 (p = 0.001). The activities of superoxide dismutase and glutathione peroxidase showed a linear increase with increasing dietary MO content, while xanthine oxidase showed a linear decrease. Total antioxidant capacity showed quadratic improvement (p = 0.035) with the dietary treatment, with the optimal dose observed at 1.25 mL/kg of diet. The inclusion of MO in the diet had a linear effect on the activity of thyroxine (p = 0.001). Therefore, supplementation of MO at a dose of 1 or 1.5 mL/kg of diet in growing rabbits'diets can improve growth and carcass traits, sustain thyroid function by supporting immunity, and reduce oxidative/inflammation pathways.

Identification of the binding proteins of organic acid metabolites by matrix thermal shift assay

Organic acid metabolites exhibit acidic properties. These metabolites serve as intermediates in major carbon metabolic pathways and are involved in several biochemical pathways, including the tricarboxylic acid (TCA) cycle and glycolysis. They also regulate cellular activity and play crucial roles in epigenetics, tumorigenesis, and cellular signal transduction. Knowledge of the binding proteins of organic acid metabolites is crucial for understanding their biological functions. However, identifying the binding proteins of these metabolites has long been a challenging task owing to the transient and weak nature of their interactions. Moreover, traditional methods are unsuitable for the structural modification of the ligands of organic acid metabolites because these metabolites have simple and similar structures. Even minor structural modifications can significantly affect protein interactions. Thermal proteome profiling (TPP) provides a promising avenue for identifying binding proteins without the need for structural modifications. This approach has been successfully applied to the identification of the binding proteins of several metabolites. In this study, we investigated the binding proteins of two TCA cycle intermediates, i.e., succinate and fumarate, and lactate, an end-product of glycolysis, using the matrix thermal shift assay (mTSA) technique. This technique involves combining single-temperature (52 ℃) TPP and dose-response curve analysis to identify ligand-binding proteins with high levels of confidence and determine the binding affinity between ligands and proteins. To this end, HeLa cells were lysed, followed by protein desalting to remove endogenous metabolites from the cell lysates. The desalted cell lysates were treated with fumarate or succinate at final concentrations of 0.004, 0.04, 0.4, and 2 mmol/L in the experimental groups or 2 mmol/L sodium chloride in the control group. Considering that the cellular concentration of lactate can be as high as 2-30 mmol/L, we then applied lactate at final concentrations of 0.2, 1, 5, 10, and 25 mmol/L in the experimental groups or 25 mmol/L sodium chloride in the control group. Using high-sensitivity mass spectrometry coupled with data-independent acquisition (DIA) quantification, we quantified 5870, 5744, and 5816 proteins in succinate, fumarate, and lactate mTSA experiments, respectively. By setting stringent cut-off values (i.e., significance of changes in protein thermal stability (p-value)<0.001 and quality of the dose-response curve fitting (square of Pearson's correlation coefficient, R2)>0.95), multiple binding proteins for these organic acid metabolites from background proteins were confidently determined. Several known binding proteins were identified, notably fumarate hydratase (FH) as a binding protein for fumarate, and α-ketoglutarate-dependent dioxygenase (FTO) as a binding protein for both fumarate and succinate. Additionally, the affinity data for the interactions between these metabolites and their binding proteins were obtained, which closely matched those reported in the literature. Interestingly, ornithine aminotransferase (OAT), which is involved in amino acid biosynthesis, and 3-mercaptopyruvate sulfurtransferase (MPST), which acts as an antioxidant in cells, were identified as lactate-binding proteins. Subsequently, an orthogonal assay technique developed in our laboratory, the solvent-induced precipitation (SIP) technique, was used to validate the mTSA results. SIP identified OAT as the top target candidate, validating the mTSA-based finding that OAT is a novel lactate-binding protein. Although MPST was not identified as a lactate-binding protein by SIP, statistical analysis of MPST in the mTSA experiments with 10 or 25 mmol/L lactate revealed that MPST is a lactate-binding protein with a high level of confidence. Peptide-level empirical Bayes t-tests combined with Fisher's exact test also supported the conclusion that MPST is a lactate-binding protein. Lactate is structurally similar to pyruvate, the known binding protein of MPST. Therefore, assuming that lactate could potentially occupy the binding site of pyruvate on MPST. Overall, the novel binding proteins identified for lactate suggest their potential involvement in amino acid synthesis and redox balance regulation.

Personalized drug screening using patient-derived organoid and its clinical relevance in gastric cancer

The efficacy of chemotherapy varies significantly among patients with gastric cancer (GC), and there is currently no effective strategy to predict chemotherapeutic outcomes. In this study, we successfully establish 57 GC patient-derived organoids (PDOs) from 73 patients with GC (78%). These organoids retain histological characteristics of their corresponding primary GC tissues. GC PDOs show varied responses to different chemotherapeutics. Through RNA sequencing, the upregulation of tumor suppression genes/pathways is identified in 5-fluorouracil (FU)- or oxaliplatin-sensitive organoids, whereas genes/pathways associated with proliferation and invasion are enriched in chemotherapy-resistant organoids. Gene expression biomarker panels, which could distinguish sensitive and resistant patients to 5-FU and oxaliplatin (area under the dose-response curve [AUC] >0.8), are identified. Moreover, the drug-response results in PDOs are validated in patient-derived organoids-based xenograft (PDOX) mice and are consistent with the actual clinical response in 91.7% (11/12) of patients with GC. Assessing chemosensitivity in PDOs can be utilized as a valuable tool for screening chemotherapeutic drugs in patients with GC.

Secondary cancer risk in six anatomical sites when using PAT, IMPT, and VMAT/IMRT radiotherapy

Background and purposeCompared to intensity modulated proton therapy (IMPT), proton arc therapy (PAT) is expected to improve dose conformality, delivery efficiency, and provide a more favorable LET distribution. Alternatively, the low-dose bath is potentially spread over larger volumes, which could impact the likelihood of developing a radiation-induced, secondary cancer (SC). The goal of this study was to evaluate this risk in several anatomical sites using newly developed commercial tools. Materials and methodsTreatment plans encompassing six anatomical sites, five patients per site, and three techniques per patient were created using RayStation. Techniques included PAT and IMPT for protons, and either volumetrically modulated radiotherapy (VMAT) or intensity modulated radiotherapy (IMRT) for photons. Risk estimates were based on the organ-equivalent dose (OED) concept using both Schneider’s mechanistic dose–response model for carcinoma induction and a linear dose–response model. ResultsWith few exceptions, mean and integral dose were lowest with PAT. For protons, the factor OEDIMPT/OEDPAT ranged from 0.7 to 1.8 with both the mechanistic and linear model, while for photons OEDphoton/OEDPAT ranged from 1.5 to 10 using the mechanistic model and 1.3 to using the linear model. A strong correlation was found between mean dose and OED for organs with significant repopulation/repair (high R value) and less cell death from single hit interactions (low α value). ConclusionBased on results from both mechanistic and linear risk models, the transition from IMPT to PAT should not substantially affect SC risk in patients treated with proton therapy. Additionally, when using Schneider’s model, the shapes of the dose–response curves can be used as a good predictor of how SC risk will respond to shifts from intermediate dose to low dose as anticipated when moving from IMPT to PAT.

Climate change in edaphic systems – Impact of salinity intrusions in terrestrial invertebrates

Amongst climate change’s impacts a major concern is salinization of soils, for example due to saltwater intrusion. The aim of the present study was to investigate in a standard field soil the impacts of soil salinity increase. The purpose was to study this in two soil invertebrates that are key model ecotoxicology test-species, Enchytraeus crypticus and Folsomia candida as surrogate representatives of the soil ecosystem. The exposure followed the standard ecotoxicity OECD (Organization for Economic Cooperation and Development) test guidelines, and the assessed endpoints were survival, reproduction and size. Exposure done in LUFA 2.2 soil, spiked with 0,0.6,1,2,3,4,5,6,8 g NaCl/kg soil dry weight (DW) for E. crypticus (21 days) and 0,0.06,0.6,1,2,3,4,5,6 g NaCl/kg soil DW for F. candida (28 days). There was a similar impact for both species in terms of survival (LC50=4.2 g NaCl/kg soil DW), whereas at the reproductive output level of F. candida (EC50=2.1 g NaCl/kg soil DW) was more sensitive than E. crypticus (EC50=2.4 g NaCl/kg soil DW). Further, size was impacted for F. candida in a monotonic dose-response curve for both adults (EC50=3.5 g NaCl/kg soil DW) and juveniles (EC50=2 g NaCl/kg soil DW), whereas for E. crypticus there was an increase in reproductive output at lower concentrations (0.6–1 g NaCl/kg soil DW). This increased reproduction was associated with a larger size of adults within the same concentration range. Considering the prediction from the climate models, the soil invertebrate community will be affected. As upper soils are likely to have the highest salinity increase due to evaporation, soil surface species, such as the collembolan tested here, are at higher risk. Negative population effects were occurring within salinity levels predicted by climate change models.

Serum copper and obesity among healthy adults in the National Health and Nutrition Examination Survey.

Copper (Cu) homeostasis are important processes in the cause of metabolic diseases, but the association between Cu and obesity remains unclear. Participants were drawn from the 2011-2016 National Health and Nutrition Examination Survey (NHANES). Weighted logistic regression assessed the associations of serum Cu concentrations (tertiles) with obesity and central obesity in individuals without comorbidities. Obesity was defined as a BMI ≥30.0 kg/m2, and central obesity was defined as a waist circumference ≥80 cm for women and ≥95 cm for men. This cross-sectional study included 1,665 adults without comorbidities, representing 24,744,034 people (mean age 35.1 years, 48.5% female). High serum Cu levels (tertile 3: ≥19.19 μmol/L) were associated with higher odds of obesity (adjusted odds ratio [OR]: 4.48, 95% CI[confidence interval]: 2.44-8.32) and central obesity (OR: 2.36, 95% CI: 1.19-4.66) compared to low serum Cu levels (tertile 1: ≤15.64 μmol/L). The dose-response curve showed a nonlinear association between Cu levels and obesity (P-nonlinear = 0.02) and a linear association with central obesity (P-nonlinear = 0.21). This study suggests that higher serum Cu levels are associated with increased odds of obesity in healthy American adults.

A Flexible Semi-automated Assay for Assessing Radiation-sensitization and Toxicity in the Mouse Intestine.

The aim of this study was to develop an enhanced intestinal toxicity assay with three outputs assessing proliferation, villi morphology and DNA damage after irradiation. Whole 5 cm jejunal lengths were collected from mice following total body x-ray irradiation (0-15 Gy) at 0-84 h. Tissues were wrapped into swirls for cryopreservation and immunohistochemically stained for EdU, CD31, and γH2AX. A semi-automated image analysis was developed for the proliferation, villi morphology, and DNA damage models. Proliferation assessed via EdU staining varied with cycles of damage repair, hyperproliferation, and homeostasis after radiation, with the time to onset of each cycle variable based on radiation dose. An analysis model evaluating the amount of proliferation per unit length of jejunum analyzed was developed, with a dose-response curve identified at 48 h post treatment. The villi length model measured the length of intact and healthy CD31-stained capillary beds between the crypts and villi tips at 3.5 days post treatment within a 0-10 Gy dose range. The DNA damage model evaluated the intensity of γH2AX staining within cellular nuclei, with a useful dose-response identified at 1 h post-radiation treatment. This assay demonstrates flexibility for assessing radiation-induced damage, with analysis of proliferation, villi length, or direct DNA damage achievable at defined time points and within useful radiation dose curves. The software-assisted image analysis allows for rapid, comprehensive, and objective data generation with an assay turnover time of days instead of weeks on samples that are representative of most of the treated jejunum.

5-hydroxytryptamine 2C/1A receptors modulate the biphasic dose response of the head twitch response and locomotor activity induced by DOM in mice.

The phenylalkylamine hallucinogen (-)-2,5-dimethoxy-4-methylamphetamine (DOM) exhibits an inverted U-shaped dose-response curve for both head twitch response (HTR) and locomotor activity in mice. Accumulated studies suggest that HTR and locomotor hyperactivity induced by DOM are mainly caused by the activation of serotonin 5-hydroxytryptamine 2A receptor (5-HT2A receptor). However, the mechanisms underlying the biphasic dose response of HTR and locomotor activity induced by DOM, particularly at high doses, remain unclear. The primary objective of this study is to investigate the modulation of 5-HT2A/2C/1A receptors in HTR and locomotor activity, while also exploring the potential receptor mechanisms underlying the biphasic dose response of DOM. In this study, we employed pharmacological methods to identify the specific 5-HT receptor subtypes responsible for mediating the biphasic dose-response effects of DOM on HTR and locomotor activity in C57BL/6J mice. The 5-HT2A receptor selective antagonist (R)-[2,3-di(methoxy)phenyl]-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (M100907) (500µg/kg, i.p.) fully blocked the HTR at every dose of DOM (0.615-10mg/kg, i.p.) in C57BL/6J mice. M100907 (50µg/kg, i.p.) decreased the locomotor hyperactivity induced by a low dose of DOM (0.625, 1.25mg/kg, i.p.), but had no effect on the locomotor hypoactivity induced by a high dose of DOM (10mg/kg) in C57BL/6J mice. The 5-HT2C antagonist 6-chloro-5-methyl-1-[(2-[2-methylpyrid-3yloxy]pyrid-5yl)carbamoyl]indoline (SB242084) (0.3, 1mg/kg, i.p.) reduced the HTR induced by a dose of 2.5mg/kg DOM, but did not affect the response to other doses. SB242084 (1mg/kg, i.p.) significantly increased the locomotor activity induced by DOM (0.615-10mg/kg, i.p.) in mice. The 5-HT1A antagonist N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]N-(2-pyridinyl) cyclohexane carboxamide maleate (WAY100635) (1mg/kg, i.p.) increased both HTR and locomotor activity induced by DOM in mice. The 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) (1mg/kg, i.p.) significantly reduced both the HTR and locomotor activity induced by DOM in mice. Additionally, pretreatment with the Gαi/o inhibitor PTX (0.25µg/mouse, i.c.v.) enhanced the HTR induced by DOM and attenuated the effect of DOM on locomotor activity in mice. Receptor subtypes 5-HT2C and 5-HT1A are implicated in the inverted U-shaped dose-response curves of HTR and locomotor activity induced by DOM in mice. The biphasic dose-response function of HTR and locomotor activity induced by DOM has different mechanisms in mice.

A Plasmodium falciparum genetic cross reveals the contributions of pfcrt and plasmepsin II/III to piperaquine drug resistance.

Piperaquine (PPQ) is widely used in combination with dihydroartemisinin as a first-line treatment against malaria. Multiple genetic drivers of PPQ resistance have been reported, including mutations in the Plasmodium falciparum chloroquine resistance transporter (pfcrt) and increased copies of plasmepsin II/III (pm2/3). We generated a cross between a Cambodia-derived multidrug-resistant KEL1/PLA1 lineage isolate (KH004) and a drug-susceptible Malawian parasite (Mal31). Mal31 harbors a wild-type (3D7-like) pfcrt allele and a single copy of pm2/3, while KH004 has a chloroquine-resistant (Dd2-like) pfcrt allele with an additional G367C substitution and multiple copies of pm2/3. We recovered 104 unique recombinant parasites and examined a targeted set of progeny representing all possible combinations of variants at pfcrt and pm2/3. We performed a detailed analysis of competitive fitness and a range of PPQ susceptibility phenotypes with these progenies, including PPQ survival assay, area under the dose response curve, and a limited point IC50. We find that inheritance of the KH004 pfcrt allele is required for reduced PPQ sensitivity, whereas copy number variation in pm2/3 further decreases susceptibility but does not confer resistance in the absence of additional mutations in pfcrt. A deep investigation of genotype-phenotype relationships demonstrates that progeny clones from experimental crosses can be used to understand the relative contributions of pfcrt, pm2/3, and parasite genetic background to a range of PPQ-related traits. Additionally, we find that the resistance phenotype associated with parasites inheriting the G367C substitution in pfcrt is consistent with previously validated PPQ resistance mutations in this transporter.IMPORTANCEResistance to piperaquine, used in combination with dihydroartemisinin, has emerged in Cambodia and threatens to spread to other malaria-endemic regions. Understanding the causal mutations of drug resistance and their impact on parasite fitness is critical for surveillance and intervention and can also reveal new avenues to limiting the evolution and spread of drug resistance. An experimental genetic cross is a powerful tool for pinpointing the genetic determinants of key drug resistance and fitness phenotypes and has the distinct advantage of quantifying the effects of naturally evolved genetic variation. Our study was strengthened since the full range of copies of KH004 pm2/3 was inherited among the progeny clones, allowing us to directly test the role of the pm2/3 copy number on resistance-related phenotypes in the context of a unique pfcrt allele. Our multigene model suggests an important role for both loci in the evolution of this multidrug-resistant parasite lineage.

Relationship of Dose and Signal Enhancement Properties of Gadoquatrane, a New Tetrameric, Macrocyclic Gadolinium-Based Contrast Agent, Compared With Gadobutrol: A Randomized Crossover Study in Healthy Adults.

To investigate the signal-enhancement properties of the tetrameric gadolinium-based contrast agent (GBCA) gadoquatrane in relation to the administered dose and compare its properties to those of a standard dose of gadobutrol, as a representative of the currently established macrocyclic GBCAs for magnetic resonance imaging. In this randomized, single-blind, 4 × 4 crossover study, 43 healthy adults (19-50 years of age) received 3 single IV injections of gadoquatrane (0.01, 0.03, and 0.06 mmol gadolinium/kg body weight) and 1 injection of gadobutrol (0.1 mmol gadolinium/kg body weight) in randomized sequence with 1-week washout periods between administrations. The relative signal enhancement (RSE) was determined in predefined areas of interest in magnetic resonance image sets of the head-neck region. RSE-vs-dose curves (dose-response curves) were established by linear regression, and comparator-equivalent doses were determined by Bayesian inverse regression analysis. Further, 3 blood samples were taken after each injection for pharmacokinetic analyses, and safety data were assessed. The RSE increased with gadoquatrane dose. A linear function adequately fitted this relationship. In line with the more than 2-fold higher r1 relaxivity of gadoquatrane per gadolinium ion, gadobutrol-equivalent RSE was achieved with gadoquatrane at less than half the gadolinium dose and less than one eighth of the molecule dose.Administration of gadoquatrane and gadobutrol resulted in very similar dose-normalized gadolinium concentrations in plasma, indicating that the pharmacokinetic profiles are essentially the same. Both contrast agents were well tolerated. Adverse events were rare and not dependent on the dose administered. Gadoquatrane has the potential to be an effective GBCA that can be used at substantially lower doses in clinical routine than the currently established macrocyclic GBCAs.

BNT12, a novel hybrid peptide of opioid and neurotensin pharmacophores, produces potent central antinociception with limited side effects

The development of multitarget opioid drugs has emerged as an attractive approach for innovative pain management with reduced side effects. In the present study, a novel hybrid peptide BNT12 containing the opioid and neurotensin (NT)-like fragments was synthesized and pharmacologically characterized. In acute radiant heat paw withdrawal test, intracerebroventricular (i.c.v.) administration of BNT12 produced potent antinociception in mice. The central antinociceptive activity of BNT12 was mainly mediated by μ-, δ-opioid receptor, neurotensin receptor type 1 (NTSR1) and 2 (NTSR2), supporting a multifunctional agonism of BNT12 in the functional assays. BNT12 also exhibited significant antinociceptive effects in spared nerve injury (SNI)-neuropathic pain, complete Freund's adjuvant (CFA)-induced inflammatory pain, acetic acid-induced visceral and formalin-induced pain after i.c.v. administration. Furthermore, BNT12 exhibited substantial reduction of acute antinociceptive tolerance, shifted the dose-response curve to the right by only 1.3-fold. It is noteworthy that BNT12 showed insignificant chronic antinociceptive tolerance at the supraspinal level. In addition, BNT12 exhibited reduced or no opioid-like side effects on conditioned place preference (CPP) response, naloxone-precipitated withdrawal response, acute hyperlocomotion, motor coordination, gastrointestinal transit, and cardiovascular responses. The present investigation demonstrated that the novel hybrid peptide BNT12 might serve as a promising analgesic candidate with limited opioid-like side effects.

Safe immunosuppression. New tool for personalized immunosuppressant treatment in renal transplantation. A case report

Background: The adjustment of immunosuppressive therapy after kidney transplantation (KT) to avoid graft rejection remains an important challenge for clinicians. It is difficult to achieve a good balance between under-immunosuppression (with an increased risk of graft rejection) and over-immunosuppression (with an increased risk of side effects) by only relying on the available information about immunosuppressive drugs (IMS). Immunobiogram® (IMBG) is a novel in vitro diagnostic test that provides clinicians with information about the patient’s sensitivity to individual IMS. Objective: To present a case report of a patient with renal transplant in the maintenance phase who presented several complications probably related to the immunosuppression during the follow-up, where the use of IMBG as complementary information helped clinicians to guide the therapeutical decision. Methods: IMBG is a first-in-class in vitro immunoassay that involves the culture of the patient peripheral blood mononuclear cells (PBMCs) in a semi-solid 3D matrix, then submitted to immune stimulation. It reveals the capacity of an IMS over a gradient to inhibit the activation of immune cells. The read-out allows the building of a dose-response curve per IMS tested, which is mathematically analyzed by a software using the key curve parameters and finally to be translated into a sensitivity map to IMS. Findings: We present a case report of a 72-year-old patient with a cadaveric donor kidney transplant receiving standard immunosuppressive treatment with mycophenolate, tacrolimus, and corticosteroids. The patient presented several episodes of infections during the follow-up (SARS-CoV2, Cytomegalovirus, spondylodisquitis by Staphylococcus aureus, and emphysematous cystitis) which were managed with different treatment adjustments such as de-escalation of mycophenolate and switching to mTOR. The information provided by the IMBG showed a lack of sensitivity to mTOR which allowed to confirm the final adjustment to a treatment with tacrolimus and corticosteroids, remaining the patient stable since then. Discussion: Despite various adjustments to the immunosuppressive therapy during the follow-up, the patient continued experiencing adverse effects that could be related to an over-immunosuppression state. The IMBG provided pharmacodynamic information that complemented the clinical and pharmacokinetic data available, facilitating the individualization of the treatment. Conclusion: The case highlights the potential of the IMBG as a complementary clinical tool for personalized treatment of kidney transplant patient management.

Single-grain luminescence and combined U-series/ESR dating of the early Upper Palaeolithic Lagar Velho Rock Shelter, Leiria, Portugal

The existing radiocarbon (14C) dating framework for Lagar Velho is broadly consistent but provides limited constraint on several geoarchaeological complexes (GCs) and does not favour detailed chronological comparisons across all sectors of the site; including the stratigraphically disconnected child burial complex in the east area and the broader archaeological sequence in the central-west area. In this study, we undertake a complementary chronological assessment of Lagar Velho Rock Shelter using single-grain quartz OSL, single-grain quartz TT-OSL and combined U-series/ESR dating of fossil teeth, with the aim of establishing more comprehensive reconstructions of archaeological events and placing the human occupation sequence in a firmer regional climatic context. Expanding on the original chronological study, we also compare the published 14C datasets against widely used sample quality indicators (i.e., organic preservation and contamination proxies) to ensure reliable comparisons with our new luminescence and combined U-series/ESR ages. Eight 14C samples pass our combined chronological and stratigraphic hygiene criteria and are included in the final dating evaluations. Ten of the twelve optical dating samples produce homogeneous equivalent dose (De) datasets indicative of suitable daylight exposure. The replicate single-grain TT-OSL De datasets exhibit more pronounced scatter, consistent with enhanced potential for insufficient bleaching of TT-OSL residual doses in some karst settings. The fossil enamel samples dated using U-series/ESR span relatively low natural dose ranges, necessitating the inclusion of maximum dose (Dmax)-adjusted dose response curve fitting and additional background noise subtraction to avoid De biases of up to 13%. Stratigraphically consistent ages (n = 26) spanning the full archaeological infill sequence are obtained using the four different dating methods, providing a robust interpretive framework and underscoring the significant role of single-grain OSL and combined U-series/ESR dating for refining early Upper Palaeolithic histories of Iberia. Bayesian modelling of the combined chronological dataset reveals a site accumulation history spanning 35,750–23,440 cal. BP and provides improved age constraints on all GCs; particularly the al, bs, tc, gs and ls deposits that were previously undated or imprecisely constrained. The age of the child burial complex is refined to 29,410–28,280 cal. BP, potentially occurring within Greenland Interstadial 4. Comparison of the modelled GC ages from the east and west-central areas enables improved temporal correlations of depositional events across stratigraphically disconnected sectors of the site. This analysis suggests that the child burial complex and the Late Gravettian occupation are statistically indistinguishable based on current dating evidence. The new dating assessments reveal three periods of human occupation at Lagar Velho: the Late Gravettian (including the shorter duration child burial event) 31.6–26.0 ka from the end of Heinrich Stadial (HS) 3 until the emergence of HS2; the Terminal Gravettian 26.5–24.5 ka coinciding with HS2; and the Middle Solutrean 25.2–23.4 ka extending from HS2 to the end of Greenland Stadial 3. The timing of these occupations during late Marine Isotope Stage (MIS) 3 and early MIS 2 highlights the significance of central western Portugal for supporting regional human populations during periods when much of Europe was experiencing extreme cold, aridity and expanded glacial coverage.

Association between dietary folate intake and the risk of osteoporosis in adults: a cross-sectional study

BackgroundIncreased intake of specific vitamins has been linked to a decreased prevalence of osteoporosis. However, the association between dietary folate intake and the risk of osteoporosis in the general population remains incompletely understood. Therefore, we aimed to determine the association between dietary folate intake and the risk of osteoporosis in the general population of the USA.MethodsIn this cross-sectional study, data from the National Health and Nutrition Examination Survey (2017–2020) were collected. Osteoporosis was considered to be indicated by a bone mineral density greater than 2.5 standard deviations below the mean of the young adult reference group. Dietary folate intake was measured by a 24-hour dietary recall. Multivariate logistic regression models and restricted cubic spline models were used.ResultsThe study included 2297 participants (mean age: 63.69 ± 0.35 years), 49.92% of whom were female. In the general population, increased dietary folate intake was directly associated with a decreased risk of osteoporosis (P for trend = 0.005). In the age > 60 years and female subgroups, folate intake was inversely associated with the risk of osteoporosis (P for trend < 0.001). The dose‒response curve suggested that this association was nonlinear (P for nonlinearity = 0.015).ConclusionsOur cross-sectional study provides initial insights into the inverse association between dietary folate intake and the risk of osteoporosis in the general U.S. population. Further research is needed to confirm these associations.

A detailed quartz and feldspar luminescence chronology for the Khonako II loess section (Southern Tajikistan)

Parts of Central Asia have been dominated by high dust flux since the start of the Quaternary. The resulting loess-palaeosol deposits are important archives for landscape and climate changes, but there is a lack of chronological data for the subaerial deposits of Southern Tajikistan. Our study presents a high-resolution luminescence chronology for the Khonako II section in the Khovaling region. Thirty-seven samples were collected from the first ∼8 m of the section, corresponding to the Holocene soil and the upper part of the Loess 1 (L1) unit. We applied quartz OSL dating to the coarse-silt fraction (40–63 μm) and polymineral pIRIR200,290 dating to the very fine sand fraction (63–90 μm). The polymineral pIRIR200,290 signal has satisfactory characteristics. The quartz dose response curve displays a non-single exponential growth above ∼100 Gy, thus, the De values were estimated by fitting a single exponential function plus linear component to the dose response data. To assess the pIRIR200,290 residual dose at deposition, we used the offset between our quartz and polymineral chronologies from Khonako II and equivalent doses measured on modern dust samples from Tajikistan and Uzbekistan. The resulting 9.2 ± 0.4 Gy dose was then subtracted from the pIRIR200,290 doses for final age calculation. The quartz OSL and pIRIR ages are then generally consistent. The obtained high-resolution luminescence chronology shows that the upper part of the L1 unit accumulated during the end of marine isotope stage (MIS) 3 and through MIS 2 (39–20 ka). The average dust accumulation rates (DAR) increased from ∼31 cm ka−1 to 41 cm ka−1 between MIS 3 and MIS 2. The DAR reached a peak of ∼71 cm ka−1 at 21 ka. A hiatus of ∼7–9 ka was then identified between the end of the last glacial maximum (LGM) and the beginning of the Holocene, and the average DAR decreased during the Holocene. Our results indicate a continuous high dust flux in the Khovaling plateau over the last 39 ka, but reworking processes and potential deflation events mark the LGM-Holocene transition. High-resolution luminescence dating studies in Tajikistan are therefore important to detect potential discontinuities in the sequences and provide an independent timescale for interpreting the climate proxies and calculating dust accumulation rates.

The impact of different strategies for modeling associations between medications at low doses and health outcomes: a simulation study and practical application to postpartum opioid use.

Pharmacoepidemiological studies commonly examine the association between drug dose and adverse health outcomes. In situations where no safe dose exists, the choice of modeling strategy can lead to identification of an apparent safe low dose range in the presence of a non-linear relationship or due to the modeling strategy forcing a linear relationship through a dose of 0. We conducted a simulation study to assess the performance of several regression approaches to model the drug dose-response curve at low doses in a setting where no safe range exists, including the use of a (1) linear dose term, (2) categorical dose term, and (3) natural cubic spline terms. Additionally, we introduce and apply an expansion of prior work related to modeling dose-response curves at low and infrequently used doses in the setting of no safe dose ("spike-at-zero" and "slab-and-spline"). Furthermore, we demonstrate and empirically assess the use of these regression strategies in a practical scenario examining the association between the dose of the initial postpartum opioid prescribed after vaginal delivery and the subsequent total dose of opioids prescribed in the entire postpartum period among a cohort of opioid-naïve women with a vaginal delivery enrolled in a State Medicaid program (2007-2014).

A novel biomimetic nanoplasmonic sensor for rapid and accurate evaluation of checkpoint inhibitor immunotherapy.

Immune checkpoint inhibitors (ICIs) emerged as promising immunotherapies for cancer treatment, harnessing the patient's immune system to fight and eliminate tumor cells. However, despite their potential and proven efficacies, checkpoint inhibitors still face important challenges such as the tumor heterogeneity and resistance mechanisms, and the complex in vitro testing, which limits their widespread applicability and implementation to treat cancer. To address these challenges, we propose a novel analytical technique utilizing biomimetic label-free nanoplasmonic biosensors for rapid and reliable screening and evaluation of checkpoint inhibitors. We have designed and fabricated a low-density nanostructured plasmonic sensor based on gold nanodisks that enables the direct formation of a functional supported lipid bilayer, which acts as an artificial cell membrane for tumor ligand immobilization. With this biomimetic scaffold, our biosensing approach provides real-time, highly sensitive analysis of immune checkpoint pathways and direct assessment of the blocking effects of monoclonal antibodies in less than 20min/test. We demonstrate the accuracy of our biomimetic sensor for the study of the programmed cell death protein 1 (PD1) checkpoint pathway, achieving a limit of detection of 6.7ng/mL for direct PD1/PD-L1 interaction monitoring. Besides, we have performed dose-response inhibition curves for an anti-PD1 monoclonal antibody, obtaining a half maximal inhibitory concentration (IC50) of 0.43nM, within the same range than those obtained with conventional techniques. Our biomimetic sensor platform combines the potential of plasmonic technologies for rapid label-free analysis with the reliability of cell-based assay in terms of ligand mobility. The biosensor is integrated in a compact user-friendly device for the straightforward implementation in biomedical and pharmaceutical laboratories.