Four-parameter Logistic Model Research Articles

A compact microfluidic laser-induced fluorescence immunoassay system using avalanche photodiode for rapid detection of alpha-fetoprotein.

Alpha-fetoprotein (AFP), commonly employed for early diagnosis of liver cancer, serves as a biomarker for cancer screening and diagnosis. Combining the high sensitivity and specificity of fluorescence immunoassay (FIA), developing a low-cost and efficient immunoassay system for AFP detection holds significant importance in disease diagnosis. In this work, we developed a miniaturized oblique laser-induced fluorescence (LIF) immunoassay system, coupled with a microfluidic PMMA/paper hybrid chip, for rapid detection of AFP. The system employed an avalanche photodiode (APD) as the detector, and implemented multi-level filtering in the excitation light channel using the dichroic mirror and optical trap. At first, we employed the Savitzky-Golay filter and baseline off-set elimination methods to denoise and normalize the original data. Then the cutoff frequency of the low-pass filter and the reverse voltage of the APD were optimized to enhance the detection sensitivity of the system. Furthermore, the effect of laser power on the fluorescence excitation efficiency was investigated, and the sampling time during the scanning process was optimized. Finally, a four-parameter logistic (4PL) model was utilized to establish the concentration-response equation for AFP. The system was capable of detecting concentrations of AFP standard solution within the range of 1-500 ng/mL, with a detection limit of 0.8 ng/mL. The entire immunoassay process could be completed within 15 min. It has an excellent potential for applications in low-cost portable diagnostic instruments for the rapid detection of biomarkers.

Ultrasensitive cortisol electrochemical immunosensor amplifying by Au single-atom nanozymes and HRP enzymes

Cortisol, a corticosteroid hormone as a primary stress hormone response to internal and external stress, has been regarded as a gold standard reliable biomarker to evaluate human mental stress. The double enzymes strategy, using nanozyme and enzyme amplifying the electrochemical signal, has been widely used to improve the performance of electrochemical biosensors. An ultra-sensitive electrochemical cortisol sensor based on Au single-atom nanozymes had been fabricated through HRP labeled anti-cortisol antibody binding with Au by Au-S bond. Based on the high catalytic activity of Au single-atom nanozymes and the high selectivity of HRP-labeled anti-cortisol antibodies, the cortisol electrochemical sensor-based Au single-atom nanozymes had an excellent response to cortisol, such as high electrochemical activity, high sensitivity, high selectivity, and wide linear range (0.15–300 ng mL−1) and low detection (0.48 pg mL−1) through the four-parameter logistic model with 95% confidence. The electrochemical cortisol sensor was used to determine the cortisol concentration of human saliva at different times.

The lupus anticoagulant titer is associated with elevated antiphosphatidylserine/prothrombin immunoglobulin-M isotype antibody levels.

Clot based assays used for lupus anticoagulant (LAC) detection are typically interpreted in a qualitative fashion and may not reflect LAC potency. In this cross-sectional study, we describe a method for quantifying the LAC titer using serial (dependent) two-fold dilutions in normal pooled plasma. Serial dilutions of 51 residual plasma samples from 50 patients were tested using the Russell's viper venom screening time (DRVVT) and activated partial thromboplastin screening time (APTT) methodologies. The measured clotting times and the corresponding dilution factors were then used to derive a four-parameter logistic model. The LAC titer for each patient was interpolated as the sample dilution that corresponds to the upper reference interval limit of the corresponding assay. Calculated APTT and DRVVT LAC titers displayed a strong linear correlation (R2 = 0.84) between each other, but not with the degree of prolongation of the APTT/DRVVT screening time in the neat undiluted samples. Using data driven partitioning, patients could be grouped into low (<10) or high (≥10) DRVVT LAC titer. There were no significant differences in anticardiolipin (aCL) or anti-beta 2 glycoprotein 1 (aB2GPI) antibody levels or prevalence of thromboembolic events between low and high LAC titer groups. In contrast, antiphosphatidylserine/prothrombin (aPS/PT) IgM antibody levels, but not IgG, were significantly higher in the high LAC titer group. The degree of prolongation of the APTT/DRVVT screening time is not correlated with the LAC titer. Only aPS/PT IgM antibodies levels were strongly correlated with the LAC titers. Additional studies are warranted to determine clinical implications of high LAC titers.

Simultaneous extraction of crocin and geniposide from gardenia fruits (Gardenia jasminoides Ellis) by probe-type ultrasound-assisted natural deep eutectic solvents and their inhibition effects on low density lipoprotein oxidation

The simultaneous extraction of crocin and geniposide from gardenia fruits (Gardenia jasminoides Ellis) was performed by integrating natural deep eutectic solvents (NADES) and ultrasound-assisted extraction (UAE). Among the eight kinds of NADES screened, choline chloride-1,2-propylene glycol was the most suitable extractant. The probe-type ultrasound-assisted NADES extraction system (pr-UAE-NADES) demonstrated higher extraction efficiency compared with plate-type ultrasound-assisted NADES extraction system (pl-UAE-NADES). Orthogonal experimental design and a modified multi-index synthetic weighted scoring method were adopted to optimize pr-UAE-NADES extraction process. The optimal extraction conditions that had a maximum synthetic weighted score of 29.46 were determined to be 25 °C for extraction temperature, 600 W for ultrasonic power, 20 min for extraction time, and 25% (w/w) for water content in NADES, leading to the maximum yields (7.39 ± 0.20 mg/g and 57.99 ± 0.91 mg/g, respectively) of crocin and geniposide. Thirty-three compounds including iridoids, carotenoids, phenolic acids, flavonoids, and triterpenes in the NADES extract were identified by LC-Q-TOF-MS2 coupled with a feature-based molecular networking workflow. The kinetics evaluation of the conjugated dienes generation on Cu2+-induced low density lipoprotein (LDL) oxidation via the four-parameter logistic regression model showed that crocin increased the lag time of LDL oxidation in a concentration-dependent manner (15 μg/mL, 30 μg/mL, 45 μg/mL) by 12.66%, 35.44%, and 73.42%, respectively. The quantitative determination for fluorescence properties alteration of the apolipoprotein B-100 exhibited that crocin effectively inhibited the fluorescence quenching of tryptophan residues and the modification of lysine residues caused by reactive aldehydes and malondialdehydes. The pr-UAE-NADES showed significant efficiency toward the simultaneous extraction of crocin and geniposide from gardenia fruits. And this study demonstrates the potential utility of gardenia fruits in developing anti-atherogenic functional food.

Exploration and analysis of a generalized one-parameter item response model with flexible link functions.

This paper primarily analyzes the one-parameter generalized logistic (1PGlogit) model, which is a generalized model containing other one-parameter item response theory (IRT) models. The essence of the 1PGlogit model is the introduction of a generalized link function that includes the probit, logit, and complementary log-log functions. By transforming different parameters, the 1PGlogit model can flexibly adjust the speed at which the item characteristic curve (ICC) approaches the upper and lower asymptote, breaking the previous constraints in one-parameter IRT models where the ICC curves were either all symmetric or all asymmetric. This allows for a more flexible way to fit data and achieve better fitting performance. We present three simulation studies, specifically designed to validate the accuracy of parameter estimation for a variety of one-parameter IRT models using the Stan program, illustrate the advantages of the 1PGlogit model over other one-parameter IRT models from a model fitting perspective, and demonstrate the effective fit of the 1PGlogit model with the three-parameter logistic (3PL) and four-parameter logistic (4PL) models. Finally, we demonstrate the good fitting performance of the 1PGlogit model through an analysis of real data.

Gaining a deeper understanding of the meaning of the carelessness parameter in the 4PL IRT model and strategies for estimating it

Three popular models are used to describe the characteristics of the test items and estimate the ability of examinees under the dichotomous IRT model, namely the one-, two-, and three-parameter logistic models. The three-item parameters are discriminating power, difficulty, and pseudo-guessing. In the development of the dichotomous IRT model, carelessness or upper asymptote parameter was proposed, which forms a four-parameter logistic (4PL) model to accommodate a condition where a high-ability examinee gives an incorrect response to a test item when he/she should be able to respond to the test item correctly. However, the carelessness parameter and the 4PL model have not been widely accepted and used due to several factors, and people’s understanding of that parameter and strategies for estimating it is still inadequate. Therefore, this study aims to shed light on ideas underlying the 4PL model, the meaning of the carelessness parameter, and strategies used to estimate that parameter based on the extant literature. The focus of this study was then extended to demonstrating practical examples of estimating item and person parameters using the 4PL model using empirical data on responses of 1,000 students from the Indonesia Open University (Universitas Terbuka) on 21 of 30 multiple-choice items on the Business English test, a paper-and-pencil test. We mainly analyzed empirical data using the ‘mirt’ package in RStudio. We present the analysis results coherently so that IRT users would have a sufficient understanding of the 4PL model and the carelessness parameter, and they can estimate item and person parameters under the 4PL model.

Reevaluation of the dietary methionine requirement of gilthead seabream (Sparus aurata) juveniles: A comparison of the efficiency of hydroxy methionine with DL-methionine as a dietary methionine source

This work aimed to estimate the dietary methionine (Met) requirement of gilthead seabream (Sparus aurata) juveniles using hydroxy methionine (OH-Met) as Met source and compare its efficiency with the DL-Methionine form (DL-Met). For that purpose, triplicate groups of fish with 17 g initial body weight were fed for 83 days with diets including increasing dietary Met levels (0.66, 0.80, 0.94, 1.08, 1.22, and 1.36% - sulfur-equivalent) using OH-Met as a Met source. Three other diets were also formulated with 0.94, 1.08, and 1.22% DL-Met, to compare OH-Met with DL-Met as dietary Met source. Cysteine was maintained at 0.54% in all the diets.Fish growth increased with dietary OH-Met inclusion up to 1.22% plateauing thereafter. No effect on feed intake was observed between fish fed the different dietary Met levels and sources. Feed and protein efficiencies increased with the increase in dietary Met level. Dietary Met source did not affect growth performance, feed utilization, nitrogen, and energy retention. Based on the four-parameter logistic model, the dietary Met requirement for maximum growth was estimated to be 1.05%. The lowest hepatic level of reduced glutathione (GSH), oxidized glutathione (GSSG), total glutathione (tGSH), and malondialdehyde (MDA) were observed in fish fed the OH-Met 1.36 diet, but the oxidative stress index (OSI) and GSH/GSSG ratio were not affected by the dietary OH-Met level. In the muscle, GSH, GSSG, OSI, MDA, and GSH/GSSG ratio were not affected by the OH-Met level, while tGSH linearly decreased with increasing dietary OH-Met level. Dietary Met source did not affect the liver and muscle oxidative stress-related parameters measured. Overall, it is concluded that the Met requirement for growth of gilthead seabream juveniles is satisfied with 1.05% Met in the diets (corresponding to 1.59% total sulfur amino acids, TSAA). This corresponds to a Met requirement of 2.3 g/16 g N (3.5 g/ 16 g N of TSAA). It is also concluded that OH-Met and DL-Met can be used interchangeably as dietary Met sources and that the liver and muscle antioxidant status of fish is not affected by the dietary Met sources tested.

Bioremediation of aniline aerofloat wastewater at extreme conditions using a novel isolate Burkholderia sp. WX-6 immobilized on biochar

Aniline aerofloat (AAF) is a refractory organic pollutant in floatation wastewater. Little information is currently available on its biodegradation. In this study, a novel AAF-degrading strain named Burkholderia sp. WX-6 was isolated from mining sludge. The strain could degrade more than 80% of AAF at different initial concentrations (100–1000 mg/L) within 72 h. AAF degrading curves were fitted well with the four-parameter logistic model (R2 >0.97), with the degrading half-life ranging from 16.39 to 35.55 h. This strain harbors metabolic pathway for complete degradation of AAF and is resistant to salt, alkali, and heavy metals. Immobilization of the strain on biochar enhanced both tolerance to extreme conditions and AAF removal, with up to 88% of AAF removal rate in simulated wastewater under alkaline (pH 9.5) or heavy metal pollution condition. In addition, the biochar-immobilized bacteria removed 59.4% of COD in the wastewater containing AAF and mixed metal ions within 144 h, significantly (P < 0.05) higher than those by free bacteria (42.6%) and biochar (48.2%) only. This work is helpful to understand AAF biodegradation mechanism and provides viable references for developing practical biotreatment technique of mining wastewater.

Abstract 207: Ex vivo modeling of multiple myeloma: A novel drug sensitivity screening platform

Abstract Multiple Myeloma (MM) is a heterogeneous malignancy characterized by abnormal clonal plasma cell infiltration in the bone marrow. It is the second most common hematologic malignancy, after non-Hodgkin’s Lymphoma. This deadly disease globally affects 1 to 5 people in every 100,000 people each year. After initial diagnosis, the 5-year survival rate is 44% because there is no curative therapy, and most patients will eventually experience relapse, and some become refractory. Large inter-patient and intra-patient genetic heterogeneity limit the identification of universal drivers of MM. However, several oncogenic dependencies such as primary events related to driver gene mutations and primary translocations can be targeted for better treatment strategies for R/R multiple myeloma. To establish a robust drug sensitivity screening platform, both MM cell lines and cryopreserved primary MM patient samples were used. MM cell lines screen with H929, MM1R and RPMI-8226 was performed to narrow down optimal concentrations of investigative test agents. During assay development, a panel of FDA approved agents for relapsed MM including panobinostat, bortezomib, melphalan, omacetaxine, and selinexor were tested. The cells were seeded at 50,000 cells per well in triplicates in a 96-well plate on day 0. The test agents were added in 9 concentrations with the top concentration being 25 uM with 2-fold dilutions. After 6- days of incubation, a luminescent cell viability assay was performed to calculate relative viability with drug treatment and IC50 was computed by fitting data using a standard four-parameter logistic model. For primary multiple myeloma samples, bone marrow aspirates were collected from Multiple Myeloma patients procured from different providers. BM mononuclear (BMMNC) cells were separated by Ficoll-Hypaque density sedimentation. Unfractionated BMMNC cells were used in these assays to preserve intra-tumor heterogeneity. Primary cells were seeded in triplicates per dose point on day 0 in Champions’ proprietary media. The test agents were also added on day 0 and were incubated for 6 days. On Day 6, the relative cell viability was measured based on Cell Titer Glo readout. The cryopreserved primary MM patient samples were viable in culture for the entire assay duration validating assay feasibility. A panel of FDA approved, standard of care drugs were tested in numerous clinically well-annotated and diverse MM patient models. Primary model selection was based on disease burden including plasma cell count from Core Biopsy and BM aspirates. Multiparametric flow cytometric immunophenotyping was also performed in these primary samples using monoclonal antibodies against CD56, CD19, CD117, CD27, CD138, and CD38. For the first time, we demonstrate the feasibility of a novel, ex-vivo drug sensitivity screening platform with cryopreserved, primary multiple myeloma cells from diverse, clinically well-annotated patient samples. Citation Format: Vaishnavi Sambandam, Sharvari Inamdar, Haoting Hsu, Brandon Walling, Paolo Schiviani, Abhay Andar, Marianna Zipeto, Michael Ritchie, Karin Abarca Heidemann, Maria Mancini. Ex vivo modeling of multiple myeloma: A novel drug sensitivity screening platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 207.

Mixture-modelling-based Bayesian MH-RM algorithm for the multidimensional 4PLM.

Several recent works have tackled the estimation issue for the unidimensional four-parameter logistic model (4PLM). Despite these efforts, the issue remains a challenge for the multidimensional 4PLM (M4PLM). Fu et al. (2021) proposed a Gibbs sampler for the M4PLM, but it is time-consuming. In this paper, a mixture-modelling-based Bayesian MH-RM (MM-MH-RM) algorithm is proposed for the M4PLM to obtain the maximum a posteriori (MAP) estimates. In a comparison of the MM-MH-RM algorithm to the original MH-RM algorithm, two simulation studies and an empirical example demonstrated that the MM-MH-RM algorithm possessed the benefits of the mixture-modelling approach and could produce more robust estimates with guaranteed convergence rates and fast computation. The MATLAB codes for the MM-MH-RM algorithm are available in the online appendix.

The apparent interferon resistance of transmitted HIV-1 is possibly a consequence of enhanced replicative fitness.

HIV-1 transmission via sexual exposure is an inefficient process. When transmission does occur, newly infected individuals are colonized by the descendants of either a single virion or a very small number of establishing virions. These transmitted founder (TF) viruses are more interferon (IFN)-resistant than chronic control (CC) viruses present 6 months after transmission. To identify the specific molecular defences that make CC viruses more susceptible to the IFN-induced 'antiviral state', we established a single pair of fluorescent TF and CC viruses and used arrayed interferon-stimulated gene (ISG) expression screening to identify candidate antiviral effectors. However, we observed a relatively uniform ISG resistance of transmitted HIV-1, and this directed us to investigate possible underlying mechanisms. Simple simulations, where we varied a single parameter, illustrated that reduced growth rate could possibly underly apparent interferon sensitivity. To examine this possibility, we closely monitored in vitro propagation of a model TF/CC pair (closely matched in replicative fitness) over a targeted range of IFN concentrations. Fitting standard four-parameter logistic growth models, in which experimental variables were regressed against growth rate and carrying capacity, to our in vitro growth curves, further highlighted that small differences in replicative growth rates could recapitulate our in vitro observations. We reasoned that if growth rate underlies apparent interferon resistance, transmitted HIV-1 would be similarly resistant to any growth rate inhibitor. Accordingly, we show that two transmitted founder HIV-1 viruses are relatively resistant to antiretroviral drugs, while their matched chronic control viruses were more sensitive. We propose that, when present, the apparent IFN resistance of transmitted HIV-1 could possibly be explained by enhanced replicative fitness, as opposed to specific resistance to individual IFN-induced defences. However, further work is required to establish how generalisable this mechanism of relative IFN resistance might be.

The Lupus Anticoagulant Titer Is Associated with Elevated Anti-Phosphatidyl-Serine/Prothrombin Antibody Levels

Background Laboratory diagnosis of the antiphospholipid antibody syndrome (APS) requires evidence of persistently positive lupus anticoagulant (LAC) activity in clot-based assays and/or the presence of medium-high titer anticardiolipin (aCL) and/or anti-beta 2 glycoprotein I (aB2GPI) antibodies. Although aCL and aB2GPI methodologies provide quantitative information about the antibody titer, LAC testing is interpreted in a qualitative fashion (presence versus absence). Objectives In this study, we describe a method for measuring the LAC titer and evaluate the relationship between the LAC titer, other antiphospholipid antibodies and clinical outcomes. Methods Residual plasma from 51 LAC-positive samples (50 unique patients) were included in this study. To estimate LAC-titers, samples were tested undiluted and subjected to serial (dependent) twofold dilutions in pooled normal plasma (PrecisionBiologic) up to a final dilution of 1:128, as appropriate. The dilute Russell's viper venom screening time (DRVVT screen, LA Check, PrecisionBiologic) and activated partial thromboplastin screening time (APTT, SynthasIL, Werfen) were measured for each dilution. A four-parameter logistic model of the dilution factor versus the clotting time was used to interpolate the LAC-titer at the upper limit of normal for both assays (APTT of 37s and DRVVT screen ratio of 1.2). Measurements of aCL, aB2GPI and anti-phosphatidylserine/prothrombin antibodies (aPS/PT) were performed using ELISA methodologies (QUANTA Lite, Werfen). Available clinical data were extracted from the electronic medical record. Continuity corrected chi-square test was used as a categorical test of association and quantile regression was used to compare medians for interval/ratio data across subgroups (Stata v17). Results There was a strong positive linear relationship between the APTT and DRVVT titers (Pearson's correlation coefficient, r=0.92, Figure 1A). There was only a moderate relationship between the APTT titer and the APTT screen in the undiluted sample (r=0.50) and almost no relationship between the DRVVT titer and the DRVVT screen ratio or DRVVT screen/confirm ratio in the undiluted sample (r=0.06 and r=0.13, respectively). Data-driven partitioning identified two DRVVT titer groups: low-titer (<10) and high-titer (≥10). Empirical cutpoint estimation (Youden) identified an APTT titer threshold of 5 for predicting the DRVVT titer group. Among patients with available clinical history, 15 of 25 (60%) low-DRVVT-titer and 16 of 20 (80%) high-DRVVT-titer patients had thromboembolic/obstetric events (p=0.26). Autoimmune diagnoses were more prevalent in the low-DRVVT-titer group (19 of 25 patients, 13 with SLE) than the high-DRVVT-titer group (8 of 20 patients, 3 with SLE, p=0.03). Comparing the low and high-DRVVT-titer groups, median aCL IgM (p=0.27), aCL IgG (p=0.99), aB2GPI IgM (p=0.72) and aB2GPI IgG (p=1.00) titers were not different. High-DRVVT-titer samples had a significantly higher median aPS/PT IgM compared to low-DRVVT-titer samples (p <0.01, Figure 1B) but there was no difference observed for IgG aPS/PT (p=0.57, Figure 1B). Conclusions APTT and DRVVT screening times are not strongly correlated with the corresponding APTT or DRVVT LAC-titer and may not indicate LAC potency. High DRVVT LAC-titers were strongly associated with IgM aPS/PT levels, but not with aCL, aB2GPI or IgG aPS/PT levels. Further research is required to evaluate the association between the LAC-titer and recurrent thrombosis in APS. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Estimating three- and four-parameter MIRT models with importance-weighted sampling enhanced variational auto-encoder.

Multidimensional Item Response Theory (MIRT) is widely used in educational and psychological assessment and evaluation. With the increasing size of modern assessment data, many existing estimation methods become computationally demanding and hence they are not scalable to big data, especially for the multidimensional three-parameter and four-parameter logistic models (i.e., M3PL and M4PL). To address this issue, we propose an importance-weighted sampling enhanced Variational Autoencoder (VAE) approach for the estimation of M3PL and M4PL. The key idea is to adopt a variational inference procedure in machine learning literature to approximate the intractable marginal likelihood, and further use importance-weighted samples to boost the trained VAE with a better log-likelihood approximation. Simulation studies are conducted to demonstrate the computational efficiency and scalability of the new algorithm in comparison to the popular alternative algorithms, i.e., Monte Carlo EM and Metropolis-Hastings Robbins-Monro methods. The good performance of the proposed method is also illustrated by a NAEP multistage testing data set.

The midpoint of cortical thinning between late childhood and early adulthood differs between individuals and brain regions: Evidence from longitudinal modelling in a 12-wave neuroimaging sample

Charting human brain maturation between childhood and adulthood is a fundamental prerequisite for understanding the rapid biological and psychological changes during human development. Two barriers have precluded the quantification of maturational trajectories: demands on data and demands on estimation. Using high-temporal resolution neuroimaging data of up to 12-waves in the HUBU cohort (N = 90, aged 7-21 years) we investigate changes in apparent cortical thickness across childhood and adolescence. Fitting a four-parameter logistic nonlinear random effects mixed model, we quantified the characteristic, s-shaped, trajectory of cortical thinning in adolescence. This approach yields biologically meaningful parameters, including the midpoint of cortical thinning (MCT), which corresponds to the age at which the cortex shows most rapid thinning - in our sample occurring, on average, at 14 years of age. These results show that, given suitable data and models, cortical maturation can be quantified with precision for each individual and brain region.

Model-Data Fit using Akaike Information Criterion (AIC), Bayesian Information Criterion (BIC), and The Sample-Size-Adjusted BIC

The study determined if the 1PL, 2PL, 3PL and 4PL item response theory models best fit the data from the 2016 NECO Mathematics objective tests. Ex-post facto design was adopted for the study. The population for the study comprised 1,022,474 candidates who enrolled and sat for June/July SSCE 2016 NECO Mathematics Examination. The sample comprised 276,338 candidates who sat for the examination in three purposively Geo political zones in Nigeria (i.e., S/West, S/East and N/West). The research instruments used for the study were Optical Marks Record Sheets for the National Examination Council (NECO) June/July 2016 SSCE Mathematics objectives items. The responses of the tests were scored dichotomously. Data collected were analyzed using 2loglikelihood chi-square. The results of the likelihood ratio test revealed that 2PL fitted the data better than 1PL was statistically significant (χ2 (59) = 820636.1, p 0.05); the 2PL model fitted the data better than the 1PL model; 3PL model fitted the data better than the 2PL model and the result showed that the 4PL model fitted the data better than the 3PL model and the Likelihood ratio test that 4PL model fitted the data better than 3PL model was statistically significant, (χ2(60)=216159.2, p0.05). The study concluded that four-parameter logistic model fitted the 2016 NECO Mathematics test items.Keywords: Akaike Information Criterion (AIC), Bayesian Information Criterion (BIC), one parameter model, two parameter model, three parameter model, four parameter model.

In vivo dose\u2013response analysis to acetylcholine: pharmacodynamic assessment by polarized reflectance spectroscopy

Transdermal iontophoresis offers an in vivo alternative to the strain-gauge model for measurement of vascular function but is limited due to lack of technical solutions for outcome assessment. The aims of this study were to, after measurement by polarized reflectance spectroscopy (PRS), use pharmacodynamic dose–response analysis on responses to different concentrations of acetylcholine (ACh); and to examine the effect of three consecutively administered iontophoretic current pulses. The vascular responses in 15 healthy volunteers to iontophorised ACh (5 concentrations, range 0.0001% to 1%, three consecutive pulses of 0.02 mA for 10 min each) were recorded using PRS. Data were fitted to a four-parameter logistic dose response model and compared. Vascular responses were quantifiable by PRS. Similar pharmacodynamic dose response curves could be generated irrespectively of the ACh concentration. Linearly increasing maximum vasodilatory responses were registered with increasing concentration of ACh. A limited linear dose effect of the concentration of ACh was seen between pulses. Polarized reflectance spectroscopy is well suited for measuring vascular responses to iontophoretically administrated ACh. The results of this study support further development of iontophoresis as a method to study vascular function and pharmacological responses in vivo.

Testing for the Equality of Variance for the Four-parameter Logistic (4PL) Model for the Detection of Vibrio cholerae DNA with Polystyrene-coacrylic Acid Composite Nanospheres

When it comes to finding the best fit of nonlinear curves to acceptable models, linear regression with least squares is the most effective technique. Because residuals (the difference between observed and predicted data) must follow a normal distribution and the data must be free of outliers and uniform variance, statistical tests are used to identify the most appropriate model for a given situation (homoscedasticity). If all of these characteristics are satisfied, the system is said to be robust. In parametric nonlinear regression, one of the numerous assumptions is that the within-group variances of the groups are all the same, which is one of several assumptions (exhibit homoscedasticity). If the variances vary from one another (show heteroscedasticity), then the model is not statistically competent to describe the data as a whole. Data on the detection of Vibrio cholerae DNA with polystyrene-coacrylic acid composite nanospheres as modelled using the nonlinear four-parameter logistic (4PL) regression was preliminary check for homogeneity of variance using the Bartlett’s and Levene’s tests. It was found that the critical value of 2 was 28.869, according to Bartlett's test findings. Excel's CHIDIST function yielded a probability of 0.389 (not significant), suggesting that the variances of the residuals did not change significantly. The p-value for Levenes's test was 0.917, indicating that there were no distinct changes between the residual variances meaning that the use of the 4-PL model in fitting the data was adequate statistically.

Detection Limit of the Four-Parameter Logistic Model for the Quantitative Detection of Serum Squamous Cell Carcinoma Antigenin Cervical Cancer Based on Surface Plasmon Resonance Biosensor

Biochemical diagnostic procedures, such as protein binding, rely on biomolecular interactions as its diagnostic modality, and as a consequence, their calibration curves are more complex. In addition, sigmoidal curves are often seen in these tests. In the event of asymmetry, a logistic (5PL) curve, or a logistic (4PL) curve, may be the best way to represent a distinctive sigmoidal relationship. It is possible that the linearization of an otherwise nonlinear connection by log transformation may result in a disruption of the error structure of the curve, and that this will have the opposite effect of decreasing or even eliminating error in the relationship. Previously, a surface plasmon resonance biosensor for the detection of squamous cell carcinoma antigen (SCCa) using nanoparticle technology was developed. However, based on the calibration curve, it conforms to the majority on sigmoidal shape curve for antibody-type sensing system. The resultant curve showed a sigmoidal calibration curve but was not modelled according to any of the sigmoidal models available. The LOD value obtained through the 4PL modelling exercise based on the classical method was 0.255 pM (95% confidence interval of 0.167 to 0.379) while the pooled standard deviation (PSD) method yielded an LOD value of 0.035 pM (95% confidence interval of 0.011 to 0.067), which indicates that the PSD method was superior.

Research Note: Development of a sandwich ELISA for determining plasma growth hormone concentrations in goose

Growth hormone (GH) is required for normal postnatal development in poultry; however, no immunoassay exists to assess its levels in geese plasma, hindering the study of endocrine regulation in this species. We developed a sandwich ELISA to determine the GH concentrations in the plasma of geese. Recombinant goose GH was produced using a eukaryotic expression system and purified for use as the reference standard in ELISA and the antigen for producing the polyclonal antibodies in rabbits. Rabbit anti-goose GH polyclonal antibody was used to coat the wells of the ELISA plate, and its biotinylated form served as the detection antibody. An avidin-conjugated horseradish peroxidase was used to bind the detection antibody and catalyze the chromogenic reaction of 3,3,5,5-tetramethylbenzidine and H2O2. A sigmoidal curve was fitted to the optical density and the log of the standard GH concentration using the four-parameter logistic model. The sensitivity of the assay was less than 0.156 ng/mL. The intra- and interassay coefficients of variation were less than 9 and 13%, respectively. The response curve of the serially diluted plasma samples from geese exhibited a good parallel relationship with that observed for the reference standards. The assay effectively detected differences in GH concentrations in plasma samples from geese at various physiological stages; thus, it will be useful for future study of their growth and metabolism.

Critical timing of weed removal in dry bean as influenced by the use of preemergence herbicides

AbstractThe critical timing of weed removal (CTWR) is the point in crop development when weed control must be initiated to prevent crop yield loss due to weed competition. A field study was conducted in 2018 and 2020 near Scottsbluff, NE, to determine how the use of preemergence herbicides affects the CTWR in dry bean. The experiment was arranged as a split plot, with herbicide treatment and weed removal timing as main and sub-plot factors, respectively. Herbicide treatments consisted of no-preemergence application, or pendimethalin (1,070 g ai ha–1) + dimethenamid-P (790 g ai ha–1) applied preemergence. Sub-plot treatments included season-long weed-free, weed removal at: V1, V3, V6, R2, and R5 dry bean growth stages, and a season-long weedy control. A four-parameter logistic model was used to estimate the impact of time of weed removal, for all response variables including dry bean yield, dry bean plants m–1 row, number of pods per plant, number of seeds per pod, and seed weight. The CTWR based on 5% yield reduction was estimated to range from the V1 growth stage [(16 d after emergence (DAE)] to the R1 growth stage (39 DAE) in the no-preemergence herbicide treatment. In the preemergence-applied treatment, the CTWR began at the R2 growth stage (47 DAE). Number of dry bean plants m–1 row was reduced in the no-preemergence treatment when weed removal was delayed beyond the R2 growth stage in the 2020 field season. The use of preemergence herbicides prevented a reduction in the number of pods per plant in 2020, and the number of seeds per pod in 2018 and 2020. In 2018, the number of pods per plant was reduced by 73% when no preemergence herbicide was applied, compared to 26% in the preemergence-applied treatment. The use of preemergence-applied soil-active herbicides in dry bean delayed the CTWR and preserved yield potential.