Standard Curve Research Articles

Head-to-head comparison of appropriate use criteria for knee arthroplasty: A multicenter cohort study

ObjectiveTo determine, in a head-to-head comparison, which of two RAND-based knee replacement appropriateness criteria is optimal based on comparison to an externally validated method of judging good versus poor outcome. DesignLongitudinal data from the Osteoarthritis Initiative (OAI) and the Multicenter Osteoarthritis Study (MOST) were combined to produce a dataset of 922 persons with knee arthroplasty, 602 of which had adequate data for RAND classification and had their surgery within one year prior to a study visit. Data were used to determine appropriateness classification (i.e., Appropriate, Inconclusive, Rarely Appropriate) using modified versions of the first-generation and second-generation Escobar system. Growth curve analyses and multivariable regression were used to compare the two systems. ResultsNeither system associated with the gold standard measure of good versus poor outcome. Distributions of appropriateness categories for the second-generation system were inconsistent with current evidence for knee arthroplasty outcome. For example, 16% of participants were classified as Appropriate and 64% as Rarely Appropriate for pain outcome. Distributions for the first-generation system aligned with current evidence. ConclusionThe first-generation modified version of the Escobar appropriateness system is superior to the newer version but neither version associated with our gold standard growth curve analyses. Both systems only differentiate between patient classification groups preoperatively and up to ten months following surgery. Reliance on appropriateness criteria to inform long-term outcome is not warranted.

Single-Cell Sensitive Colorimetric pH Detection Based on Microscope Ratiometric Grayscale.

Rapid and accurate identification of the intracellular pH is critical in the field of biomedicine. In this work, we effectively identified and quantified the intracellular pH and its distribution at the single-cell level using an image sensor based on an ordinary bright-field optical microscope that divided the cell staining images into their red (R) and blue (B) channels. The grayscale of the R and B channels was subjected to a ratiometric operation to generate ratiometric grayscale cell images of the microscope. A standard curve of pH against ratiometric grayscale curve was then obtained by incubating HeLa cells at pH 6.00-7.60 in a high concentration K+ ion buffer solution containing nigericin for obtaining certain intracellular pH values. A good correlation was evidenced between pH and the ratiometric grayscale of the R and B channels in the pH range of 6.00-7.60. Subsequently, the intracellular pH value of the A549 cells under the experimental conditions was measured to be 7.22 ± 0.01 by the method. Furthermore, the changes in the intracellular pH of HeLa cells stimulated with hydrogen peroxide were sensitively monitored, which demonstrated the applicability of the method. Due to its ease of use, the developed colorimetric microscopy pH detection and monitoring method provide prospects for pH-related single-cell studies.

Wastewater monitoring of SARS-CoV-2 gene for COVID-19 epidemiological surveillance in Tucumán, Argentina.

Wastewater-based epidemiology provides temporal and spatial information about the health status of a population. The objective of this study was to analyze and report the epidemiological dynamics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the province of Tucumán, Argentina during the second and third waves of coronavirus disease 2019 (COVID-19) between April 2021 and March 2022. The study aimed to quantify SARS-CoV-2 RNA in wastewater, correlating it with clinically reported COVID-19 cases. Wastewater samples (n = 72) were collected from 16 sampling points located in three cities of Tucumán (San Miguel de Tucumán, Yerba Buena y Banda del Río Salí). Detection of viral nucleocapsid markers (N1 gene) was carried out using one-step reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Viral loads were determined for each positive sample using a standard curve. A positive correlation (p < 0.05) was observed between viral load (copies/mL) and the clinically confirmed COVID-19 cases reported at specific sampling points in San Miguel de Tucumán (SP4, SP7, and SP8) in both months, May and June. Indeed, the high viral load concurred with the peaks of COVID-19 cases. This method allowed us to follow the behavior of SARS-CoV-2 infection during epidemic outbreaks. Thus, wastewater monitoring is a valuable epidemiological indicator that enables the anticipation of increases in COVID-19 cases and tracking the progress of the pandemic. SARS-CoV-2 genome-based surveillance should be implemented as a routine practice to prepare for any future surge in infections.

A Quantitative Method to Distinguish Cytosolic from Endosome-Trapped Cell-Penetrating Peptides.

Cell-penetrating peptides are known to penetrate cells through endocytosis and translocation. The two pathways are hardly distinguished in current cell assays. We developed a reliable, simple and robust method to distinguish and quantify independently the two routes. The assay requires (DABCYL) 4-(dimethylaminoazo)benzene-4-carboxylic acid- and (CF) carboxyfluorescein-labeled peptides. When the labeled peptide is intact, the fluorescence signal is weak thanks to the dark quenching property of DABCYL. A 10-fold higher fluorescence signal is measured when the labeled peptide is degraded. By referring to a standard fluorescent curve according to the concentration of the hydrolyzed peptide, we have access to the internalized peptide quantity. Therefore, cell lysis after internalization permits to determine the total quantity of intracellular peptide. The molecular state of the internalized peptide (intact or degraded), depends on its location in cells (cytosol vs endo-lysosomes), and can be blocked by boiling cells. This boiling step results indeed in denaturation and inhibition of the cellular enzymes. The advantage of this method is the possibility to quantify translocation at 37 °C and to compare it to the 4 °C condition, where all endocytosis processes are inhibited. We found that ranking of the translocation efficacy is DABCYL-R6-(ϵCF)K≫DABCYL-R4-(ϵCF)K≥CF-R9.

70 Determination of phosphorus release of Microtech phytase in nursery pig diets

Abstract A total of 320 barrows (DNA 241 × 600; initially 10.7 ± 0.21 kg) were used in a 21-d growth trial to determine the available P (aP) release curve for Microtech phytase (Guangdong VTR Bio-Tech Co, Ltd. Zhuhai, Guangdong, China). At approximately 21 d of age, pigs were weaned and randomly allocated to pens and fed a common diet for 18 d and then fed a P depletion diet (0.11% aP) for 3 d. On d 21 post-weaning, considered d 0 of the study, pigs were blocked by average pen body weight (BW) and randomly allotted to 1 of 8 dietary treatments with 5 pigs per pen and 8 pens per treatment. Dietary treatments were derived from a single basal diet and ingredients including phytase, monocalcium P, limestone, and sand were added to create the treatment diets. Treatments include 3 diets with increasing aP (0.11, 0.19 and 0.27%) using monocalcium phosphate that were used to develop the standard curve and 5 diets with 0.11% aP and increasing phytase (250, 500, 1,000, 1,500 and 2,000 FTU/kg) that were used to develop the aP release curve. Pigs were weighed weekly and feed disappearance was recorded per pen for the calculation of average daily gain (ADG), average daily feed intake (ADFI) and gain to feed ratio (G:F). At the conclusion of the experiment, 1 pig, closest to the mean weight of each pen, was euthanized for bone analysis. The right fibula, rib, and metacarpal were collected for the determination of bone density, bone ash weight, and bone ash percentage. Pigs fed diets with increasing aP from inorganic P had improved (quadratic, P ≤ 0.05) final BW, ADG, and G:F (Table). Pigs fed diets with increasing phytase had linearly increased (P &amp;lt; 0.01) final BW and ADFI and quadratically increased (P ≤ 0.04) ADG and G:F. Bone density, bone ash weight, and bone ash percentage increased linearly (P &amp;lt; 0.01) with increasing aP from inorganic P and increasing phytase. Results were used to determine release values for Microtech phytase for each response criteria as shown in the table. The aP release quadratically increased (P ≤ 0.02) with increasing phytase when growth performance was used as response criteria and linearly increased (P &amp;lt; 0.01) when bone characteristics were used as response criteria. The available P release curve developed for Microtech phytase for percentage bone ash using combined data generated from all three bones is: aP = (0.868 × FTU/kg) ÷ (9,599.511 + FTU/kg).

Direct Extraction and Determination of Free Nicotine in Cigarette Smoke.

The accurate determination of the free nicotine content in cigarette smoke is crucial for assessing cigarette quality, studying harm and addiction, and reducing tar levels. Currently, the determination of free nicotine in tobacco products primarily relies on methods such as pH calculation, nuclear magnetic resonance (NMR) spectroscopy, headspace solid-phase microextraction (HS-SPME), and traditional solvent extraction. However, these methods have limitations that restrict their widespread application. In this study, the free nicotine in cigarette smoke was directly extracted by using cyclohexane according to the traditional solvent extraction method and detected via gas chromatography-mass spectrometry. Compared with the traditional two-phase solvent extraction, our experimental method is easy to execute and eliminates the influence of aqueous solutions on the original distribution of nicotine in cigarette smoke particulate matter. Furthermore, the presence of protonated nicotine in tobacco does not affect the determination. Compared with HS-SPME and NMR spectroscopy, our approach, which involves solvent extraction followed by chromatographic separation and instrumental detection, offers simplicity, improved precision, better detection limits, and reduced interference during the instrumental detection stage. The standard addition recoveries in the conducted experiment ranged from 96.2% to 102.5%. The limit of detection was 2.8 μg/cig, and the correlation coefficient (R2) for the quadratic regression of the standard curve exceeded 0.999. The relative standard deviation for parallel samples was between 1.7% and 3.4% (n = 5), fully meeting the requirements for the determination of free nicotine in cigarette smoke. Analysis of cigarette samples from 38 commercially available brands revealed that the content of free nicotine ranged from 0.376 to 0.716 mg/cig, with an average of 0.540 mg/cig, and free nicotine accounted for 39.1%-88.8% of the total nicotine content.

Evaluation of a miniature mass spectrometer based point-of-care-test method for direct analysis of amlodipine and benazepril in whole blood

A miniature mass spectrometer (mMS) based point-of-care testing (POCT) method was evaluated for on-site detecting the hypertension drugs, amlodipine and benazepril. The instrument parameters, including voltage, ISO1, ISO2, and CID, were optimized, under which the target compounds could be well detected in MS2. When these two drugs were injected simultaneously, the mutual ionization inhibition and mutual reduction between amlodipine and benazepril were evaluated. This phenomenon was severe on the precursor ions but had a small impact on the product ions, thus making this POCT method suitable for analysis using product ions. Finally, the method was validated and applied. The blood samples from patients were tested one hour after oral administration of the drugs (20 mg), and the benazepril was quantitatively analyzed using a standard curve, with detected concentrations ranging from 190.6 to 210 μg L−1 and a relative standard deviation (RSD) of 8.6 %. In summary, amlodipine has low sensitivity and can only be detected at higher concentrations, while benazepril has high sensitivity, good linearity, and even meets semi-quantitative requirements. The research results of this study are of great clinical significance for monitoring blood drug concentrations during hypertension medication, predicting drug efficacy, and customizing individualized medication plans.

A model of heterogeneous undercooled liquid and glass accounting for temperature-dependent nonexponentiality and enthalpy fluctuation.

Dynamic heterogeneity is a fundamental characteristic of glasses and undercooled liquids. The heterogeneous nature causes some of the key features of systems' dynamics such as the temperature dependence of nonexponentiality and spatial enthalpy fluctuations. Commonly used phenomenological models such as Tool-Narayanaswamy-Moynihan (TNM) and Kovacs-Aklonis-Hutchinson-Ramos fail to fully capture this phenomenon. Here we propose a model that can predict the temperature-dependent nonexponential behavior observed in glass-forming liquids and glasses by fitting standard differential scanning calorimetry curves. This model extends the TNM framework of structural relaxation by introducing a distribution of equilibrium fictive temperature (Tfe) that accounts for heterogeneity in the undercooled liquid. This distribution is then frozen at the glass transition to account for the heterogeneous nature of the glass dynamics. The nonexponentiality parameter βKWW is obtained as a function of temperature by fitting the Kohlrauch-Williams-Watts (KWW) equation to the calculated relaxation function for various organic and inorganic undercooled liquids and glasses. The calculated temperature dependent βKWW shows good agreement with the experimental ones. We successfully model the relaxation dynamics far from equilibrium for two silicate systems that the TNM model fails to describe, confirming that temperature dependent nonexponentiality is necessary to fully describe these dynamics. The model also simulates the fluctuation of fictive temperature δTf during isothermal annealing with good qualitative agreement with the evolution of enthalpy fluctuation reported in the literature. We find that the evolution of enthalpy fluctuation during isothermal annealing heavily depends on the cooling rate, a dependence that was not previously emphasized.

Isolation, Characterization, and Degradation Efficiency Testing of a Paclobutrazol-degrading Bacterium: Deinococcus arenae

Objective: This study aimed to isolate and characterize a bacterium capable of degrading the pesticide paclobutrazol. The strain was obtained from soil enriched with paclobutrazol, and its degradation ability was assessed using gas chromatography (GC). Methods: A single strain was isolated from the soil solution of Ophiopogon japonicus, a plant exposed to excessive paclobutrazol spraying. Initial isolation utilized dilution and plating methods, followed by separation and purification through the streak plate technique. BLAST analysis revealed the strain's highest similarity to Deinococcus arenae SA1, with 100% homology after removing excess sequences. Subsequently, the degradation efficiency of this bacterium on paclobutrazol was evaluated by gas chromatography. Results: Quantification using the external standard method yielded a standard curve equation of y=0.0977x-0.412, with a correlation coefficient (R²) of 0.99842. Testing at different time showed that the Deinococcus arenae achieved a degradation rate of 2.49% after three days of degrading paclobutrazol. After five days, the degradation rate increased to 17.58%, reaching 38.66% after seven days, 42.20% after nine days, and 43.87% after eleven days. Conclusion: The Deinococcus arenae strain exhibited a significant degradation effect on paclobutrazol. This finding suggests potential for developing microbial agents to degrade paclobutrazol, promoting sustainable practices in agriculture.

Correction of the Effect of Humic Acids on the Fluorescence Detection of Polycyclic Aromatic Hydrocarbons by Combination Spectroscopy.

Polycyclic aromatic hydrocarbons (PAHs) are widely distributed in soil and water, but fluorescence spectroscopy for PAHs is often interfered with organic matter in the environment. The aim of this paper is to evaluate a correction method using combined spectral technology in an environment where humic acids and PAHs coexist. In the present work, humic acids and benzo[ghi]perylene were analyzed in various concentrations using fluorescence and near-infrared (NIR) spectroscopy from single and mixed samples. The NIR prediction model of humic acids in mixed samples was established based on synergy interval partial least squares, and the standard curve of fluorescence spectra for humic acids was established at 478 nm (characteristic wavelength of benzo[ghi]perylene). The fluorescence intensity of humic acids in the mixed sample was predicted from the content derived from the NIR spectra. The final correction was carried out by their exclusion from the fluorescence of the mixture at the same wavelength. The corrected fluorescence intensity was linearly correlated with the concentration of benzo[ghi]perylene with R2 = 0.8362, while R2 = 0.3538 before correction. These results give a new insight into the calibration modeling of the combined spectral method.

Computational Modeling of Sodium Fluorescein Disposition in Ex Vivo, Machine-Perfused Livers with Ischemia-Reperfusion Injury

Rationale: Quantitation of bile formation machinery activity through biliary sodium fluorescein (SF) clearance is useful for assessing liver viability. Studies have shown that the bile-to-plasma SF ratio is inversely correlated with warm ischemia time (WIT) in rats, which is associated with ischemia-reperfusion injury (IRI)-induced intracellular internalization of the multidrug resistance protein 2 (MRP2) transporter (doi: 10.1152/ajpgi.00038.2022). Computational modeling studies by Monti et al. have demonstrated the sensitivity of biliary SF clearance to MRP2 transporter activity in vivo (doi: 10.48550/arXiv.2302.05511). In vivo experimentation is pertinent for establishing physiological variations occurring during IRI, but transplantation often requires a period of ex vivo machine perfusion (MP). The effects of total hepatectomy, WIT, cold ischemia time (CIT), and MP on biliary function remain unclear. We aim to develop a model for quantitative interpretation of biliary SF clearance kinetics in MP livers to estimate parameters descriptive of the dominant processes determining hepatic SF uptake kinetics in MP livers subjected to different periods of WIT. We hypothesize that increasing WIT will lead to a decrease in the maximal transport velocity (Vmax) parameter for MRP2, which is reflective of decreased biliary SF clearance and diminished liver viability. Methods: Using established protocols (doi: 10.3389/frtra.2023.1215182), livers (n = 3-5) were obtained from anesthetized, male, Sprague-Dawley rats and subjected to 0-, 10-, 20-, or 30-min WIT through ligation of the portal vein and hepatic artery. Livers were removed and subjected to 3 hours of CIT on ice. Livers were attached to a custom normothermic MP system, 0.4 mg/kg SF were added to the MP system’s perfusate-containing reservoir, and SF fluorescence measurements in perfusate and bile were obtained and converted to concentration measurements using standard curves. We modified the liver-centric model described in Monti et al. to account for MP by replacing the liver input functions with ordinary differential equations for SF and its conjugate SF glucuronide (SFG) kinetics in the MP system’s reservoir. The model also accounts for three liver regions (sinusoid, hepatocytes, bile) and major processes, including hepatic transporter kinetics and SF metabolism into SFG, which govern SF dynamics in different model regions. We fit the model solutions using a pseudo-Monte Carlo parameter estimation strategy to each of the datasets by allowing the Vmax parameters to vary, while others were fixed to physiologic values. Results: Time course data for IRI conditions showed a modest decrease in biliary SF with 10- and 20-min WIT and a marked change in biliary SF kinetics with 30-min WIT when compared to control. The MP model was fit to each dataset individually and the model was able to provide a good fit to each dataset. Comparison of Vmax parameters estimated from fitting showed a decrease in MRP2’s Vmax with increasing WIT relative to control. Sensitivity analysis supported our finding by demonstrating that MRP2’s Vmax­ was the most sensitive model parameter. Conclusions: We modified an existing in vivo biliary SF clearance model to account for MP livers and fit the new model to biliary SF clearance data for ex vivo MP livers with varying amounts of WIT. We found that the new model was able to fit biliary SF clearance data for ex vivo MP livers and that the Vmax for MRP2 decreased during MP with increasing WIT. Funded in part by 1F30AI179084-01 to C.M. and NSF DMS 2153387 to R.D. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

The calibration device research for assessing the impact exposure level of rubber balls in architectural acoustics

In order to accurately measure the impact exposure level of rubber balls used in architectural acoustics, a calibration device is designed based on a force sensor, signal analyzer, programmable stepper motor and Infinite Impulse Response (IIR) filter. A weighing sensor with a range of 200 kg is selected to match the range of impact forces of rubber balls. The sensor is installed on a specially designed metal base plate of a rubber ball measuring fixture and connected to the signal analyzer. The rubber ball is dropped from a predetermined height in a free-fall manner to impact the sensor, generating an impact waveform. The force exposure level is then calculated by using software algorithms built into the device. The accuracy and reliability of the calibration device are evaluated by conducting repeated measurements on rubber balls and comparing the results with a known standard force exposure level curve. The experimental results demonstrate that the calibration device can consistently and stably measure the force exposure level of rubber balls, showing consistency with the known standard values. However, due to limitations such as a limited variety of measured objects and insufficient measurement repetitions, further research will be conducted to increase the number of tested rubber balls and enhance the device’s capabilities. This will provide technical support for the further development and application of the device.

Using Students’ Smartphones as Tools to Determine Total Carbohydrates in Beverages

Smartphones are used universally in many areas due to their advanced features. Smartphones are also widely used by teachers and students in education. This research aims to design low-cost and easy experiments for biology lessons using smartphones. Colorimetric measurement was made with a smartphone. Mobile applications were used to read color change values. The amount of sugar in beverages was calculated with the students. They could create standard curves, measure with a smartphone, organize the data, and determine sugar concentration. It was determined that the students had positive thoughts about the activity. Self-confidence results indicated that the activity had a positive effect on students’ self-confidence. Although there was a decrease in the mean scores of difficulty, it was revealed that the mean scores were over the “moderate” level. As a result, it can be said that the activity may be easily practiced by teachers and students in biology lessons.

Prediction of Wells Performance and Producing Life Applying Production Decline Analysis

Analyzing the production data became a common method in modern reservoir engineering practice, which is used to dynamically indicate the reservoir's characteristics, predict long-term performance, and determine the reservoir's criteria through daily production analysis. Input data required to analyze production data is production rate and sometimes flow pressure (if any). The decline curve analysis method analyzes well production data to history matching and forecasts the future well performance. The decline curve analysis (DCA) method is a non-linear regression method for production history matching and prediction. In this study, we used the DCA method to analyze eight oil wells' production data produced from the Mishrif reservoir in an Iraqi oil field. This work dealt with the prediction of the reservoir’s performance and future production rate using the DCA method tool integrated with IPM- MBAL software. Production decline rate trend analysis was performed by generating standard curves utilizing exponential, hyperbolic, and harmonic decline model equations based on historical production data. Afterward, a production forecast model was built for production to 2025. The producing life for each well at the economic flow rate (300 bbl/Day) is estimated. Results from the production forecast showed that the exponential and hyperbolic decline models yielded 54.057 MMSTB and 80.73 MMSTB, respectively. The decline rates for hyperbolic and exponential models till 2025 were estimated as 4614.25 STB/Day and 1153.1STB/Day.

Present and Future Applications of Digital PCR in Infectious Diseases Diagnosis.

Infectious diseases account for about 3 million deaths per year. The advent of molecular techniques has led to an enormous improvement in their diagnosis, both in terms of sensitivity and specificity and in terms of the speed with which a clinically useful result can be obtained. Digital PCR, or 3rd generation PCR, is based on a series of technical modifications that result in more sensitive techniques, more resistant to the action of inhibitors and capable of direct quantification without the need for standard curves. This review presents the main applications that have been developed for the diagnosis of viral, bacterial, and parasitic infections and the potential prospects for the clinical use of this technology.

AIE-labeled fluorescent polystyrene nanoplastics for quantitative analysis in macrophages uptake

Nanoplastics have attracted significant attention due to their harmful effects on the environment and organisms. However, accurately detecting and quantifying nanoplastics that are ingested by organisms poses a challenge. Conventional organic fluorescent-labeled nanoplastics have reduced labeling efficiency due to aggregation-caused quenching (ACQ), leading to artifacts caused by leakage of fluorescent molecules. To address these limitations, this study utilized AIE fluorescence for internally labeling polystyrene nanoparticles. AIE-active molecules do not emit fluorescence in dilute solutions, thus preventing accuracy issues associated with fluorescence leakage. Three model plastics (TPE@PS) of varying particle sizes were synthesized via the swelling method. The preparation parameters were optimized, followed by stability tests conducted on TPE@PS. Fluorescence microscopy imaging revealed significant uptake of TPE@PS by RAW246.7 cells. A successful standard curve of fluorescence intensity-concentration was established successfully and utilized to accurately quantify the amount of TPE@PS that was ingested by RAW246.7 cells. The results indicate a consistent increase with increasing concentration in TPE@PS uptake across all particle sizes during the fixed exposure duration of 48 h. Furthermore, RAW264.7 cells exhibited a higher propensity to uptake smaller TPE@PS particles. Flow cytometry was also employed to validated the suitability of TPE@PS as a model nanoplastic for tracing purposes.

A comparison of free amino nitrogen and yeast-assimilable nitrogen measurement methods for use in alcoholic fermentation of whey

The continued efficient use of whey as a fermentable substrate to produce alcoholic beverages will benefit from improving the measurement of key components relevant to the fermentability of whey. One component of fermentation that is not well studied in whey is the concentration of fermentable nitrogen, either as free amino nitrogen (FAN) or yeast-assimilable nitrogen (YAN). Fermentable nitrogen in media is essential for yeast cells to replicate. Insufficient concentrations of FAN or YAN to support the growing yeast population can result in sluggish or "stuck" fermentations. We evaluated 3 common methods of fermentable nitrogen determination and compared them for use in whey fermentation systems, based on ninhydrin, nitrogen by o-phthalaldehyde (NOPA), and formaldehyde pH (Sørensen titration) values. Each measurement method was evaluated independently using standard addition curves and compared for overall accuracy using paired t-tests (α = 0.05). Although the formaldehyde pH method showed high precision, it did not measure nitrogen accurately, as it overestimated YAN in whey by up to 6 times relative to other tests. The ninhydrin and NOPA methods both showed accuracy for fermentable nitrogen determination in whey based on analysis of standardized nitrogen sources. We concluded that either the ninhydrin FAN or NOPA YAN method may be used to determine the fermentable nitrogen content in whey. This is expected to improve the ability of producers to use whey as a fermentation medium.

Construction and validation of a predictive model for major adverse cardiovascular events in the long term after percutaneous coronary intervention in patients with acute ST-segment elevation myocardial infarction.

Construction of a prediction model to predict the risk of major adverse cardiovascular events (MACE) in the long term after percutaneous coronary intervention (PCI) in patients with acute ST-segment elevation myocardial infarction (STEMI). Retrospective analysis of STEMI patients treated with PCI from April 2018 to April 2021 in Fuyang People's Hospital. Lasso regression was used to screen the risk factors for the first occurrence of MACE in patients, and multifactorial logistic regression analysis was used to construct a prediction model. The efficacy was evaluated by area under the ROC curve (AUC), Hosmer-Lemeshow deviance test, calibration curve, clinical decision curve (DCA) and clinical impact curve (CIC). Logistic regression results showed that hypertension, diabetes mellitus, left main plus three branches lesion, estimated glomerular filtration rate and medication adherence were influential factors in the occurrence of distant MACE after PCI in STEMI patients ( P < 0.05). The AUC was 0.849 in the modeling group and 0.724 in the validation group; the calibration curve had a good fit to the standard curve, and the result of the Hosmer-Lemeshow test of deviance was x 2 = 7.742 ( P = 0. 459); the DCA and the CIC indicated that the predictive model could provide a better net clinical benefit for STEMI patients. A prediction model constructed from a total of five predictor variables, namely hypertension, diabetes, left main + three branches lesions, eGFR and medication adherence, can be used to assess the long-term prognosis after PCI in STEMI patients and help in early risk stratification of patients.

Decreased IL-1 β Secretion as a Potential Predictor of Tuberculosis Recurrence in Individuals Diagnosed with HIV.

Background: The mechanisms of the formation of immunological competence against tuberculosis (TB), and especially those associated with HIV co-infection, remain poorly understood. However, there is an urgent need for risk recurrence predictive biomarkers, as well as for predictors of successful treatment outcomes. The goal of the study was to identify possible immunological markers of TB recurrence in individuals with HIV/TB co-infection. Methods: The plasma levels of IFN-γ, TNF-α, IL-10, and IL-1β (cytokines which play important roles in the immune activation and protection against Mycobacterium tuberculosis) were measured using ELISA EIA-BEST kits. The cytokine concentrations were determined using a standard curve obtained with the standards provided by the manufacturer of each kit. Results: A total of 211 individuals were enrolled in the study as follows: 62 patients with HIV/TB co-infection, 52 with HIV monoinfection, 52 with TB monoinfection, and 45 healthy donors. Out of the 62 patients with HIV/TB, 75.8% (47) of patients were newly diagnosed with HIV and TB, and 24.2% (15) displayed recurrent TB and were newly diagnosed with HIV. Decreased levels of IFN-γ, TNF-α, and IL-10 were observed in patients with HIV/TB when compared with HIV and TB patients. However, there was no difference in IFN-γ, TNF-α, or IL-10 secretion between both HIV/TB groups. At the same time, an almost 4-fold decrease in Il-1β levels was detected in the HIV/TB group with TB recurrence when compared with the HIV/TB group (p = 0.0001); a 2.8-fold decrease when compared with HIV patients (p = 0.001); and a 2.2-fold decrease with newly diagnosed TB patients (p = 0.001). Conclusions: Significantly decreased Il-1β levels in HIV/TB patients' cohort with secondary TB indicate that this cytokine can be a potential biomarker of TB recurrence.

Investigating the Presence of Rotavirus in Wastewater Samples of Bhopal Region, India, by Utilizing Droplet Digital Polymerase Chain Reaction.

Rotavirus-induced viral gastroenteritis outbreaks result in over two million hospitalizations globally yearly. Wastewater-based epidemiology (WBE) has emerged as a crucial tool for detecting and monitoring viral outbreaks. The adoption of WBE has been instrumental in the early detection and surveillance of such viral outbreaks, providing a non-invasive method to assess public health. This study aims to utilize droplet digital polymerase chain reaction (ddPCR) technology to detect and quantify Rotavirus in wastewater samples collected from the Bhopal region of India, thereby contributing to the understanding and management of viral gastroenteritis outbreaks through environmental surveillance. In this study, we used ddPCR to detect and quantify Rotavirus in wastewater samples collected from the Bhopal region of India. We monitored its viral presence in municipal sewage treatment plants bi-weekly using an advanced ddPCR assay. Targeting the rotavirus non-structural protein 3 (NSP-3) region with custom primers and TaqMan probes, we detected virus concentration employing polyethylene glycol (PEG). Following RNA isolation, complementary DNA (cDNA) synthesis, and ddPCR analysis, our novel method eliminated standard curve dependence, propelling virus research and treatment forward. Out of the 42 samples collected, a 16.60% positivity rate was observed, indicating a moderate presence of Rotavirus in Bhopal. The wastewater treatment plants (WWTP) attached to a hospital exhibited a 42.85% positivity rate, indicating the need for targeted monitoring. Leveraging ddPCR, precise quantification of rotavirus concentrations (ranging from 0.75 to 28.9 copies/µL) facilitated understanding and supported effective remediation. This study emphasizes the importance of vigilant wastewater surveillance, especially in WWTPs with higher rotavirus prevalence. The significance of ddPCR in comparison to conventional and real-time PCR lies in its superior sensitivity and specificity in detecting and quantifying positive samples. Furthermore, it can identify positive samples even in the smallest quantities without the need for a standard curve to evaluate. This makes ddPCR a valuable tool for accurate and precise detection and quantification of samples.