Reliable Screening Research Articles

Machine learning models for screening clinically significant nephrolithiasis in overweight and obese populations.

Our aim is to build and evaluate models to screen for clinically significant nephrolithiasis in overweight and obesity populations using machine learning (ML) methodologies and simple health checkup clinical and urine parameters easily obtained in clinics. We developed ML models to screen for clinically significant nephrolithiasis (kidney stone > 2mm) in overweight and obese populations (body mass index, BMI ≥ 25kg/m2) using gender, age, BMI, gout, diabetes mellitus, estimated glomerular filtration rate, bacteriuria, urine pH, urine red blood cell counts, and urine specific gravity. The data were collected from hospitals in Kaohsiung, Taiwan between 2012 and 2021. Of the 2928 subjects we enrolled, 1148 (39.21%) had clinically significant nephrolithiasis and 1780 (60.79%) did not. The testing dataset consisted of data collected from 574 subjects, 235 (40.94%) with clinically significant nephrolithiasis and 339 (59.06%) without. One model had a testing area under curve of 0.965 (95% CI, 0.9506-0.9794), a sensitivity of 0.860 (95% CI, 0.8152-0.9040), a specificity of 0.947 (95% CI, 0.9230-0.9708), a positive predictive value of 0.918 (95% CI, 0.8820-0.9544), and negative predictive value of 0.907 (95% CI, 0.8756-0.9371). This ML-based model was found able to effectively distinguish the overweight and obese subjects with clinically significant nephrolithiasis from those without. We believe that such a model can serve as an easily accessible and reliable screening tool for nephrolithiasis in overweight and obesity populations and make possible early intervention such as lifestyle modifications and medication for prevention stone complications.

Sensitive bioluminescence imaging of cryptococcosis in Galleria mellonella improves antifungal screening under in vivo conditions

ABSTRACT Cryptococcus neoformans is an environmental yeast that primarily affects immunocompromised individuals, causing respiratory infections and life-threatening meningoencephalitis. Treatment is complicated by limited antifungal options, with concerns such as adverse effects, dose-limiting toxicity, blood–brain barrier permeability, and resistance development, emphasizing the critical need to optimize and expand current treatment options against invasive cryptococcosis. Galleria mellonella larvae have been introduced as an ethical intermediate for in vivo testing, bridging the gap between in vitro antifungal screening and mouse studies. However, current infection readouts in G. mellonella are indirect, insensitive, or invasive, which hampers the full potential of the model. To address the absence of a reliable non-invasive method for tracking infection, we longitudinally quantified the cryptococcal burden in G. mellonella using bioluminescence imaging (BLI). After infection with firefly luciferase-expressing C. neoformans, the resulting bioluminescence signal was quantitatively validated using colony-forming unit analysis. Longitudinal comparison of BLI to health and survival analysis revealed increased sensitivity of BLI in discriminating cryptococcal burden during early infection. Furthermore, BLI improved the detection of treatment efficacy using first-line antifungals, thereby benchmarking this model for antifungal testing. In conclusion, we introduced BLI as a real-time, quantitative readout of cryptococcal burden in G. mellonella over time, enabling more sensitive and reliable antifungal screening.

White blood cells scattergram as a valuable tool for COVID-19 screening: A multicentric study.

New tools have been developed to distinguish the COVID-19 diagnosis from other viral infections presenting similar symptomatology and mitigate the lack of sensitivity of molecular testing. We previously identified a specific "sandglass" aspect on the white blood cells (WBC) scattergram of COVID-19 patients, as a highly reliable COVID-19 screening test (sensitivity: 85.9%, specificity: 83.5% and positive predictive value: 94.3%). We then decided to validate our previous data in a multicentric study. This retrospective study involved 817 patients with flu-like illness, among 20 centers, using the same CBC instrument (XN analyzer, SYSMEX, Japan). After training, one specialist per center independently evaluated, under the same conditions, the presence of the "sandglass" aspect of the WDF scattergram, likely representing plasmacytoid lymphocytes. Overall, this approach showed sensitivity: 59.0%, specificity: 72.9% and positive predictive value: 77.7%. Sensitivity improved with subgroup analysis, including in patients with lymphopenia (65.2%), patients presenting symptoms for more than 5 days (72.3%) and in patients with ARDS (70.1%). COVID-19 patients with larger plasmacytoid lymphocyte cluster (>15 cells) more often have severe outcomes (70% vs. 15% in the control group). Our findings confirm that the WBC scattergram analysis could be added to a diagnostic algorithm for screening and quickly categorizing symptomatic patients as either COVID-19 probable or improbable, especially during COVID-19 resurgence and overlapping with future influenza epidemics. The observed large size of the plasmacytoid lymphocytes cluster appears to be a hallmark of COVID-19 patients and was indicative of a severe outcome. Furthers studies are ongoing to evaluate the value of the new hematological parameters in combination with WDF analysis.

Psychometric testing and the predictive validity of the Postpartum Depression Predictors Inventory-Revised (PDPI-R): A longitudinal study with Turkish women

ObjectiveThe aim of this study was to investigate the validity and reliability of the prenatal and postnatal versions of the Postpartum Depression Predictors Inventory-Revised (PDPI-R) and to examine the predictive validity of PDPI-R in Turkish women, considering two gold standards to determine postpartum depression (PPD). MethodsThis prospective longitudinal study was conducted between August 2021 and September 2023. A total of 301 pregnant women participated in the study. Participants completed the PDPI-R during the third trimester of pregnancy (T1) and at 4 weeks postpartum (T2). At T2, participants also completed the Edinburgh Postnatal Depression Scale (EPDS), and women were interviewed using the Structured Clinical Interview for DSM-IV Disorders. ResultsThe prenatal version of the PDPI-R predicted PPD with 64% (R:0.64) accuracy on the basis of the EPDS and 78% accuracy (R:0.78) according to DSM IV criteria. The postnatal version of the PDPI-R predicted PPD with 71% (R:0.71) accuracy on the basis of the EPDS and 81% accuracy (R:0.781) based on DSM IV criteria. The cut-off points exhibited the highest sensitivity and specificity values at 8.5 for the prenatal version and 10.5 for the postnatal version. ConclusionsThe PDPI-R is a valid and reliable screening tool for identifying Turkish women at high risk of developing PPD and for estimating the psychosocial risk associated with PPD.

Knowledge, attitude, and practice of lung cancer screening and associated factors among high-risk population in Lanzhou, China: A cross-sectional study.

This study aimed to assess the knowledge, attitude, and practice (KAP) of high-risk populations toward lung cancer screening in Lanzhou, China. Using convenience sampling, this cross-sectional study enrolled outpatients at high-risk for lung cancer at Lanzhou University Second Hospital between November 2022 and March 2023. An anonymous, self-administered online questionnaire was distributed to each participant via the Sojump website (https://www.wjx.cn/), comprising 40 items to collect demographic information and evaluate KAP toward lung cancer screening. The analyses were descriptive. A total of 577 participants (average age of 61.8 ± 7.1 years; 306 males) were included in the study. The participants' scores for KAP were 4.9 ± 2.2, 27.4 ± 3.0, and 7.0 ± 2.1, respectively. Participants with occupational exposure had significantly lower knowledge score (3.3 ± 2.4 vs 5.2 ± 2.1, P < .001), and practice score (5.6 ± 2.4 vs 7.3 ± 1.9, P < .001) than those without occupational exposure. Participants with smoking or passive smoking history had significantly higher attitude scores (27.6 ± 2.9 vs 25.8 ± 3.2, P < .001) and practice scores (7.1 ± 2.0 vs 6.5 ± 2.5, P = .014) than those without smoking history. A total of 360 (62.4%) participants endorsed the doctors' counseling on lung cancer screening, and 355 (61.5%) participants were willing to have screening for lung cancer as doctors advised. The study revealed that 390 (67.6%) participants identified low-dose computed tomography as the appropriate method for lung cancer screening, while 356 (61.7%) participants believed that X-rays were a reliable screening method for lung cancer. However, 365 (63.3%) participants thought that the treatment outcomes for early and late-diagnosed lung cancer were the same. Additionally, 416 (72.10%) participants believed that annual lung cancer CT scanning is unnecessary. On the other hand, 339 (58.8%) participants expressed concerns about exposure to radiation from CT scans, while 349 (60.5%) participants were worried about the cost of lung cancer screening. Only 142 (24.6%) participants reported having undergone annual lung cancer screening. The high-risk population had limited knowledge and insufficient attitude and practice toward lung cancer screening in Lanzhou, China.

Evaluating cross-reactivity of new psychoactive substances (NPS) in human whole blood by enzyme-linked immunosorbent assay (ELISA).

Due to the increase in the use of novel psychoactive substances (NPS) and their overall prevalence, it is important to have effective and reliable screening technologies to detect NPS in biological matrices. Enzyme-linked immunosorbent assays (ELISA) are among the most popular screening methods. To evaluate the effectiveness of ELISA for NPS detection, five subclasses of NPS (novel synthetic opioids, fentanyl analogs, stimulants, benzodiazepines and hallucinogens) were evaluated in whole blood for their cross-reactivity on commercially available ELISA kits. A variety of novel synthetic opioids were tested at concentrations of 1-80 ng/mL and 50-2000 ng/mL and demonstrated no cross-reactivity to a morphine ELISA plate at either concentration range. Fentanyl analogs were tested at concentrations ranging from 0.01 to 1 ng/mL and had cross-reactivities ranging from 8% to 178% on the fentanyl ELISA kit used. Both para-chloro fentanyl (178%) and acryl fentanyl (164%) showed cross-reactivities well above that of fentanyl. Novel stimulants were tested at concentrations of 0.5-40 ng/mL and 20-2,000 ng/mL. 4-Fluoroamphetamine was the only novel stimulant with cross-reactivity (3,354%) to the amphetamine ELISA plate. Novel benzodiazepines were tested at concentrations of 1-40 ng/mL on a benzodiazepine plate. Cross-reactivities ranged from 36.1% to 263%, with desalkylflurazepam having the highest cross-reactivity. Finally, novel hallucinogens were tested at concentrations of 0.5-10 ng/mL on a phencyclidine (PCP) ELISA plate, which produced no cross-reactivity and then with 10-1,000 ng/mL, which gave results from 56.6% to 151%. Both hydroxy-PCP (151%) and chloro-PCP (137%) showed cross-reactivities above that of PCP. This research has demonstrated the utility of using ELISA-based screening for novel benzodiazepines, hallucinogens and for fentanyl analogs; however, there is limited application and risk of false-negative results for the other drug classes due to low or non-existent cross-reactivities.

Screening of CPT1A-Targeting Lipid Metabolism Modulators Using Mitochondrial Membrane Chromatography.

Carnitine palmitoyltransferase 1A (CPT1A), which resides on the mitochondrial outer membrane, serves as the rate-limiting enzyme of fatty acid β-oxidation. Identifying the compounds targeting CPT1A warrants a promising candidate for modulating lipid metabolism. In this study, we developed a CPT1A-overexpressed mitochondrial membrane chromatography (MMC) to screen the compounds with affinity for CPT1A. Cells overexpressing CPT1A were cultured, and subsequently, their mitochondrial membrane was isolated and immobilized on amino-silica gel cross-linked by glutaraldehyde. After packing the mitochondrial membrane column, retention components of MMC were performed with LC/MS, whose analytic peaks provided structural information on compounds that might interact with mitochondrial membrane proteins. With the newly developed MMC-LC/MS approach, several Chinese traditional medicine extracts, such as Scutellariae Radix and Polygoni Cuspidati Rhizoma et Radix (PCRR), were analyzed. Five noteworthy compounds, baicalin, baicalein, wogonoside, wogonin, and resveratrol, were identified as enhancers of CPT1A enzyme activity, with resveratrol being a new agonist for CPT1A. The study suggests that MMC serves as a reliable screening system for efficiently identifying modulators targeting CPT1A from complex extracts.

Dual branch deep learning network for detection and stage grading of diabetic retinopathy

Diabetic retinopathy is a severe complication of diabetes that can lead to permanent blindness if not treated promptly. Early and accurate diagnosis of the disease is essential for successful treatment. This paper introduces a deep learning method for the detection and stage grading of diabetic retinopathy, using a single fundus retinal image. Our model utilizes transfer learning, employing two state-of-the-art pre-trained models as feature extractors and fine-tuning them on a new dataset. The proposed model is trained on a large multi-center dataset, including the APTOS 2019 dataset, obtained from publicly available sources. It achieves remarkable performance in diabetic retinopathy detection and stage classification on the APTOS 2019, outperforming the established literature. For binary classification, the proposed approach achieves an accuracy of 98.50%, a sensitivity of 99.46%, and a specificity of 97.51%. In stage grading, it achieves a quadratic weighted kappa of 93.00%, an accuracy of 89.60%, a sensitivity of 89.60%, and a specificity of 97.72%. The proposed approach serves as a reliable screening and stage grading tool for diabetic retinopathy, offering significant potential to enhance clinical decision-making and patient care.

Three-in-one lateral flow aptasensor based on magnetic nanoparticles for on-site detection of ochratoxin A in Astragalus membranaceus

The prevalence and significant health risks associated with mycotoxin contamination in traditional Chinese medicines underscore the need for on-site detection methods. The complexity of matrices necessitates sample pretreatment before analysis, leading to prolonged analysis times and intricate procedures. In this study, we employed aptamer-modified magnetic nanoparticles (aptamer-MNPs) as labels and devised a three-in-one lateral flow aptasensor for on-site detection of ochratoxin A (OTA) in Astragalus membranaceus. By integrating aptamer-MNPs that serve in recognition, separation, and signal amplification roles, we streamlined purification, enrichment, and detection within a single aptasensor in 30 minutes. The developed aptasensor demonstrated high sensitivity (0.053 ng/mL), excellent specificity, and satisfactory accuracy. Analyzing twelve actual samples revealed three as OTA-positive, aligning well with HPLC analysis results. These results showcase the potential of the lateral flow aptasensor as a cost-effective, reliable, and robust on-site screening technique for trace-level mycotoxins in complex matrices.

TTLL12 has a potential oncogenic activity, suppression of ligation of nitrotyrosine to the C-terminus of detyrosinated α-tubulin, that can be overcome by molecules identified by screening a compound library.

Tubulin tyrosine ligase 12 (TTLL12) is a promising target for therapeutic intervention since it has been implicated in tumour progression, the innate immune response to viral infection, ciliogenesis and abnormal cell division. It is the most mysterious of a fourteen-member TTL/TTLL family, since, although it is the topmost conserved in evolution, it does not have predicted enzymatic activities. TTLL12 seems to act as a pseudo-enzyme that modulates various processes indirectly. Given the need to target its functions, we initially set out to identify a property of TTLL12 that could be used to develop a reliable high-throughput screening assay. We discovered that TTLL12 suppresses the cell toxicity of nitrotyrosine (3-nitrotyrosine) and its ligation to the C-terminus of detyrosinated α-tubulin (abbreviated to ligated-nitrotyrosine). Nitrotyrosine is produced by oxidative stress and is associated with cancer progression. Ligation of nitrotyrosine has been postulated to be a check-point induced by excessive cell stress. We found that the cytotoxicities of nitrotyrosine and tubulin poisons are independent of one another, suggesting that drugs that increase nitrotyrosination could be complementary to current tubulin-directed therapeutics. TTLL12 suppression of nitrotyrosination of α-tubulin was used to develop a robust cell-based ELISA assay that detects increased nitrotyrosination in cells that overexpress TTLL12 We adapted it to a high throughput format and used it to screen a 10,000 molecule World Biological Diversity SETTM collection of low-molecular weight molecules. Two molecules were identified that robustly activate nitrotyrosine ligation at 1 μM concentration. This is the pioneer screen for molecules that modulate nitrotyrosination of α-tubulin. The molecules from the screen will be useful for the study of TTLL12, as well as leads for the development of drugs to treat cancer and other pathologies that involve nitrotyrosination.

Novel plasma microRNA expression features in diagnostic use for Epstein-Barr virus-associated febrile diseases

BackgroundEpstein-Barr virus (EBV) is widely infected in humans and causes various diseases. Among them, microRNAs of EBV play a key role in the progression of EBV-associated febrile diseases. There're few specific indicators for rapid differential diagnosis of various febrile diseases associated with EBV, and the lack of more reliable screening methods with high diagnostic utility has led to spaces for improvement in the accurate diagnosis and efficient treatment of relevant patients, making EBV infection a complicated clinical problem. With recent advances in plasma microRNA testing, the apparent presence of EBV microRNAs in plasma can help screen for EBV infection. The gene networks targeted by these microRNAs can also indicate potential biomarkers of EBV-associated febrile diseases. This study aimed to identify some novel miRNAs as potential biomarkers for early diagnosis of respectively EBV-associated febrile diseases. Materials and methodsA total of 110 participants were recruited for this task. First, we performed high-throughput sequencing and preliminary PCR validation of differentially expressed miRNAs in 15 participants with EBV-associated fever (divided into 5 groups, common EBV carriers, infectious mononucleosis (IM) and chronic active EBV infection (CAEBV), EBV-associated Hemophagocytic Lymphohistiocytosis (EBV-HLH), and 3 healthy individuals). After a comprehensive analysis, 10 miRNAs with abnormal expression were screened, and then qRT-PCR was performed in the rest of 95 participants to detect the validation of miRNAs expression in plasma samples. Thereafter, we further investigated their potential for clinical application in EBV-related febrile diseases by using a combination of Gene Ontology analysis, Kyoto Encyclopedia of Genes and Genomes pathway analysis, and Protein-protein interaction network analysis. ResultsThrough identification and detailed analysis of the obtained data, we found significant differences in the expression of Hsa-miR-320d, EBV-miR-BART22, and EBV-miR-BART2-3p in blood samples from patients with different EBV-related febrile diseases. We found that the expression levels of Hsa-miR-320d, EBV-miR-BART22, and EBV-miR-BART2-3p in plasma are indicative of determining different disease types of EBV-related febrile diseases, while EBV-miR-BART22 and EBV-miR-BART2-3p may be potential therapeutic targets. ConclusionThe expression levels of Hsa-miR-320d, EBV-miR-BART22, and EBV-miR-BART2-3p suggest that they may be used as transcriptional features for early differential diagnosis of EBV-related febrile diseases, and EBV-miR-BART22 and EBV-miR-BART2-3p may be potential therapeutic targets.

Cervical Squamous Cell Carcinoma Diagnosis by FTIR Microspectroscopy.

Cervical cancer was considered the fourth most common cancer worldwide in 2020. In order to reduce mortality, an early diagnosis of the tumor is required. Currently, this type of cancer occurs mostly in developing countries due to the lack of vaccination and screening against the Human Papillomavirus. Thus, there is an urgent clinical need for new methods aiming at a reliable screening and an early diagnosis of precancerous and cancerous cervical lesions. Vibrational spectroscopy has provided very good results regarding the diagnosis of various tumors, particularly using Fourier transform infrared microspectroscopy, which has proved to be a promising complement to the currently used histopathological methods of cancer diagnosis. This spectroscopic technique was applied to the analysis of cryopreserved human cervical tissue samples, both squamous cell carcinoma (SCC) and non-cancer samples. A dedicated Support Vector Machine classification model was constructed in order to categorize the samples into either normal or malignant and was subsequently validated by cross-validation, with an accuracy higher than 90%.

Rapid antibiotic screening based on E. coli apoptosis using a potentiometric sensor array

Phenotypic antimicrobial susceptibility testing enables reliable antibiotic screening but requires multiple strategies to identify each phenotypic change induced by different bactericidal mechanisms. Bacteria apoptosis with typical phenotypic features has never been explored for antibiotic screening. Herein, we developed an antibiotic screening method based on the measurement of antibiotic-induced phosphatidylserine (PS) exposure of apoptotic bacteria. Phosphatidylserine externalization of E. coli that can be widely used as an apoptosis marker for antibiotics with different antibacterial mechanisms was explored. A positively charged PS-binding peptide was immobilized on magnetic beads (MBs) to recognize and capture apoptotic E. coli with PS externalization. Apoptotic E. coli binding led to the charge or charge density change of MBs-peptide, resulting in a potential change on a magneto-controlled polymeric membrane potentiometric sensor. Based on the detection of apoptotic E. coli killed by antibiotics, antibiotic screening for different classes of antibiotics and silver nanoparticles was achieved within 1.5 h using a potentiometric sensor array. This approach enables sensitive, general, and time-saving antibiotic screening, and may open up a new path for antibiotic susceptibility testing.

Advanced 3D In Vitro Lung Fibrosis Models: Contemporary Status, Clinical Uptake, and Prospective Outlooks.

Fibrosis has been characterized as a global health problem and ranks as one of the primary causes of organ dysfunction. Currently, there is no cure for pulmonary fibrosis, and limited therapeutic options are available due to an inadequate understanding of the disease pathogenesis. The absence of advanced in vitro models replicating dynamic temporal changes observed in the tissue with the progression of the disease is a significant impediment in the development of novel antifibrotic treatments, which has motivated research on tissue-mimetic three-dimensional (3D) models. In this review, we summarize emerging trends in preparing advanced lung models to recapitulate biochemical and biomechanical processes associated with lung fibrogenesis. We begin by describing the importance of in vivo studies and highlighting the often poor correlation between preclinical research and clinical outcomes and the limitations of conventional cell culture in accurately simulating the 3D tissue microenvironment. Rapid advancement in biomaterials, biofabrication, biomicrofluidics, and related bioengineering techniques are enabling the preparation of in vitro models to reproduce the epithelium structure and operate as reliable drug screening strategies for precise prediction. Improving and understanding these model systems is necessary to find the cross-talks between growing cells and the stage at which myofibroblasts differentiate. These advanced models allow us to utilize the knowledge and identify, characterize, and hand pick medicines beneficial to the human community. The challenges of the current approaches, along with the opportunities for further research with potential for translation in this field, are presented toward developing novel treatments for pulmonary fibrosis.

The equivalence of psychodynamic therapy and cognitive behavioral therapy for depressive disorders in adults: A meta-analytic review.

Meta-analyses on the relative efficacy of psychodynamic psychotherapy (PDT) and cognitive behavioral therapy (CBT) for depressive disorders are limited by heterogeneity in diagnostic samples and comparators and a lack of equivalence testing. We addressed this through a meta-analytic test of the equivalence of manualized PDT and CBT in treating adults with depressive disorders as determined by diagnostic interviews. Sensitivity analyses evaluated the impact of pretreatment differences, mixed diagnostic samples, author allegiance, study quality, year of publication and outliers on findings. A comprehensive literature search across multiple databases using reliable screening methods identified nine randomized controlled trials directly comparing manualized PDT and CBT for diagnosed depressive disorders in adults. Following pre-registration, we employed random effect models for our meta-analyses and two one-sided test procedures for equivalence testing. Independent raters determined that all studies were of adequate quality. Immediately posttreatment, depressive symptoms were statistically equivalent across PDT and CBT (k = 9; g = -0.11, 90% confidence interval [90% CI]: -0.24 to 0.02, pequivalence = .048, pNHST = .212, I2 = 32.7). At follow-up, the longest time point within a year, depressive symptoms were neither statistically equivalent nor statistically different (k = 6; g = -0.16, 90% CI: -0.31 to -0.02, pequivalence = .184, pNHST = .126, I2 = 0.00). The efficacy of manualized PDT is equal to manualized CBT immediately at posttreatment for depressive disorders in the adult general population. Nevertheless, insufficient data exists to reach a conclusion regarding equivalence at follow-up.

Rapid and low-cost screening for single and combined effects of drought and heat stress on the morpho-physiological traits of African eggplant (Solanum aethiopicum) germplasm.

Drought and heat are two stresses that often occur together and may pose significant risks to crops in future climates. However, the combined effects of these two stressors have received less attention than single-stressor investigations. This study used a rapid and straightforward phenotyping method to quantify the variation in 128 African eggplant genotype responses to drought, heat, and the combined effects of heat and drought at the seedling stage. The study found that the morphophysiological traits varied significantly among the 128 eggplants, highlighting variation in response to abiotic stresses. Broad-sense heritability was high (> 0.60) for chlorophyll content, plant biomass and performance index, electrolyte leakage, and total leaf area. Positive and significant relationships existed between biomass and photosynthetic parameters, but a negative association existed between electrolyte leakage and morpho-physiological traits. The plants underwent more significant stress when drought and heat stress were imposed concurrently than under single stresses, with the impact of drought on the plants being more detrimental than heat. There were antagonistic effects on the morphophysiology of the eggplants when heat and drought stress were applied together. Resilient genotypes such as RV100503, RV100501, JAMBA, LOC3, RV100164, RV100169, LOC 3, RV100483, GH5155, RV100430, GH1087, GH1087*, RV100388, RV100387, RV100391 maintained high relative water content, low electrolyte leakage, high Fv/Fm ratio and performance index, and increased biomass production under abiotic stress conditions. The antagonistic interactions between heat and drought observed here may be retained or enhanced during several stress combinations typical of plants' environments and must be factored into efforts to develop climate change-resilient crops. This paper demonstrates improvised climate chambers for high throughput, reliable, rapid, and cost-effective screening for heat and drought and combined stress tolerance in plants.

Mismatch between subjective and objective dysautonomia

Autonomic symptom questionnaires are frequently used to assess dysautonomia. It is unknown whether subjective dysautonomia obtained from autonomic questionnaires correlates with objective dysautonomia measured by quantitative autonomic testing. The objective of our study was to determine correlations between subjective and objective measures of dysautonomia. This was a retrospective cross-sectional study conducted at Brigham and Women’s Faulkner Hospital Autonomic Laboratory between 2017 and 2023 evaluating the patients who completed autonomic testing. Analyses included validated autonomic questionnaires [Survey of Autonomic Symptoms (SAS), Composite Autonomic Symptom Score 31 (Compass-31)] and standardized autonomic tests (Valsalva maneuver, deep breathing, sudomotor, and tilt test). The autonomic testing results were graded by a Quantitative scale for grading of cardiovascular reflexes, sudomotor tests and skin biopsies (QASAT), and Composite Autonomic Severity Score (CASS). Autonomic testing, QASAT, CASS, and SAS were obtained in 2627 patients, and Compass-31 in 564 patients. The correlation was strong between subjective instruments (SAS vs. Compass-31, r = 0.74, p < 0.001) and between objective instruments (QASAT vs. CASS, r = 0.81, p < 0.001). There were no correlations between SAS and QASAT nor between Compass-31 and CASS. There continued to be no correlations between subjective and objective instruments for selected diagnoses (post-acute sequelae of COVID-19, n = 61; postural tachycardia syndrome, 211; peripheral autonomic neuropathy, 463; myalgic encephalomyelitis/chronic fatigue syndrome, 95; preload failure, 120; post-treatment Lyme disease syndrome, 163; hypermobile Ehlers-Danlos syndrome, 213; neurogenic orthostatic hypotension, 86; diabetes type II, 71, mast cell activation syndrome, 172; hereditary alpha tryptasemia, 45). The lack of correlation between subjective and objective instruments highlights the limitations of the commonly used questionnaires with some patients overestimating and some underestimating true autonomic deficit. The diagnosis-independent subjective–objective mismatch further signifies the unmet need for reliable screening surveys. Patients who overestimate the symptom burden may represent a population with idiosyncratic autonomic-like symptomatology, which needs further study. At this time, the use of autonomic questionnaires as a replacement of autonomic testing cannot be recommended.

High-Throughput Screening Techniques for the Selection of Thermostable Enzymes.

The acquisition of a thermostable enzyme is an indispensable prerequisite for its successful implementation in industrial applications and the development of novel functionalities. Various protein engineering approaches, including rational design, semirational design, and directed evolution, have been employed to enhance thermostability. However, all of these approaches require sensitive and reliable high-throughput screening (HTS) technologies to efficiently and rapidly identify variants with improved properties. While numerous reviews focus on modification strategies for enhancing enzyme thermostability, there is a dearth of literature reviewing HTS methods specifically aimed at this objective. Herein, we present a comprehensive overview of various HTS methods utilized for modifying enzyme thermostability across different screening platforms. Additionally, we highlight significant recent examples that demonstrate the successful application of these methods. Furthermore, we address the technical challenges associated with HTS technologies used for screening thermostable enzyme variants and discuss valuable perspectives to promote further advancements in this field. This review serves as an authoritative reference source offering theoretical support for selecting appropriate screening strategies tailored to specific enzymes with the aim of improving their thermostability.

Constructing Tumor Organoid-like Tissue for Reliable Drug Screening Using Liver-Decellularized Extracellular Matrix Scaffolds.

The interplay between cells and their microenvironments plays a pivotal role in in vitro drug screening. Creating an environment that faithfully mimics the conditions of tumor cells within organ tissues is essential for enhancing the relevance of drug screening to real-world clinical scenarios. In our research, we utilized chemical decellularization techniques to engineer liver-decellularized extracellular matrix (L-dECM) scaffolds. These scaffolds were subsequently recellularized with HepG2 cells to establish a tumor organoid-like tissue model. Compared to the conventional tissue culture plate (TCP) culture, the tumor organoid-like tissue model demonstrated a remarkable enhancement in HepG2 cell growth, leading to increased levels of albumin secretion and urea synthesis. Additionally, our results revealed that, within a 3-day time frame, the cytotoxicity of doxorubicin (DOX) against cells cultured in the tumor organoid-like tissue model was notably reduced when compared to cells grown on TCPs. In contrast, there was no significant difference in the cytotoxicity of two compounds, triptolide and honokiol, both derived from traditional Chinese medicine, between TCP culture and the tumor organoid-like tissue culture, indicating a lack of substantial drug resistance. Western blotting assays further confirmed our findings by revealing elevated expressions of E-cadherin and vimentin proteins, which are closely associated with the epithelial-mesenchymal transition (EMT). These results underscored that the tumor organoid-like tissue model effectively promoted the EMT process in HepG2 cells. Moreover, we identified that triptolide and honokiol possess the capacity to reverse the EMT process in HepG2 cells, whereas DOX did not exhibit a significant effect. In light of these findings, the tumor organoid-like tissue model stands as a valuable predictive platform for screening antitumor agents and investigating the dynamics of the EMT process in tumor cells.

Illiterate Addenbrooke's Cognitive Examination-III in Three Indian Languages: An Adaptation and Validation Study.

Literacy is an important factor that predicts cognitive performance. Existing cognitive screening tools are validated only in educated populations and are not appropriate for older adults with little or no education leading to poor performance on these tests and eventually leading to misdiagnosis. This challenge for clinicians necessitates a screening tool suitable for illiterate or low-literate older individuals. The objective was to adapt and validate Addenbrooke's Cognitive Examination-III (ACE-III) for screening general cognitive functions in illiterate and low-literate older populations in the Indian context in three languages. The Indian illiterate ACE-III was systematically adapted by modifying the original items of the Indian literate ACE-III to assess the cognitive functions of illiterates and low-literates with the consensus of an expert panel of professionals working in the area of dementia and related disorders. A total of 180 illiterate or low-literate participants (84 healthy-controls, 50 with dementia, and 46 with mild cognitive impairment [MCI]) were recruited from three different centers speaking Bengali, Hindi, and Kannada to validate the adapted version. The optimal cut-off score for illiterate ACE-III to distinguish controls from dementia in all 3 languages was 75. The optimal cut-off scores in distinguishing between controls and MCI ranged from 79 to 82, with a sensitivity ranging from 93% to 99% and a specificity ranging from 72% to 99%. The test is found to have good psychometric properties and is a reliable cognitive screening tool for identifying dementia and MCI in older adults with low educational backgrounds in the Indian context.