Correlation R2 Research Articles

In vitro evaluation of a new viscoelastometry-based point-of-care analyzer.

The VCM is a point-of-care analyzer using a new viscoelastometry technique for rapid assessment of hemostasis on fresh whole blood. Its characteristics would make it suitable for use in austere environments. The purpose of this study was to evaluate the VCM in terms of repeatability, reproducibility and interanalyzer correlation, reference values in our population, correlation with standard coagulation assays and platelet count, correlation with the TEG5000 analyzer and resistance to stress conditions mimicking an austere environment. Repeatability, reproducibility, and interanalyzer correlation were performed on quality control samples (n = 10). Reference values were determined from blood donor samples (n = 60). Correlations with standard biological assays were assessed from ICU patients (n = 30) and blood donors (n = 60) samples. Correlation with the TEG5000 was assessed from blood donor samples. Evaluation of vibration resistance was performed on blood donor (n = 5) and quality control (n = 5) samples. The CVs for repeatability and reproducibility ranged from 0% to 11%. Interanalyzer correlation found correlation coefficients (r2) ranging from 0.927 to 0.997. Our reference values were consistent with those provided by the manufacturer. No robust correlation was found with conventional coagulation tests. The correlation with the TEG5000 was excellent with r2 ranging from 0.75 to 0.92. Resistance to stress conditions was excellent. The VCM analyzer is a reliable, easy-to-use instrument that correlates well with the TEG5000. Despite some logistical constraints, the results suggest that it can be used in austere environments. Further studies are required before its implementation.

Modification of the coffee industry sludge-based adsorbent using acid treatment for the removal of tartrazine dye from aqueous solution

The massive amount of industrial sludge produced annually poses an environmental concern, and improper sludge disposal may harm the environment. To address this issue, sludge has been converted into a cost-effective adsorbent for treating wastewater containing dyes. This study aims to modify the coffee sludge-based adsorbent using various treatment methods for tartrazine dye removal. The raw sludge was modified through physical treatment (PT) via oven drying and calcination, chemical treatment (CT) using phosphoric acid and nitric acid, and physicochemical treatment (PCT) method. The modified sludge adsorbents were characterised using Micromeritics ASAP 2020 for nitrogen adsorption analysis. Subsequent batch adsorption experiments were performed using the best-modified adsorbent of PCTS-HNO3 prepared by physicochemical treatment method with nitric acid to assess the effect of various factors, including initial pH (3-11), adsorbent dosage (0.05-0.25 g/L), and initial concentration of tartrazine dye (25-100 mg/L), with a contact time of 2 hours. The best process conditions were achieved at 0.15 g/L of adsorbent dosage, initial pH of 3 and initial concentration of 50 mg/L with almost 100% dye removal. The characterization results revealed that the BET surface area of the modified adsorbent was significantly increased from 363.32 m2/g of RS to 599.64 m2/g for PCTS-HNO3 adsorbent. The Langmuir isotherm model provided an excellent fit to the adsorption isotherm data, with correlation coefficients R2 ≥ 0.99 and a maximum adsorption capacity (qmax) of 99.01 mg/g. The kinetic study observed that the adsorption process adheres remarkably well to the pseudo-second-order kinetic model, signifying chemisorption adsorption. The results of this study indicate that the modified coffee sludge-based adsorbent proves to be a viable and cost-efficient solution for eliminating tartrazine dye from aqueous solutions.

Method Development and Validation for Simultaneous Estimation of Lamivudine and Raltegravir in Pure Drug form by RP-HPLC

A new, simple, accurate, and economical method was developed for simultaneous estimation of Lamivudine and Raltegravir by RP-HPLC. The chromatographic separation was achieved through Phenomenex C18 250mm × 4.6mm, 5µm as stationary phase and mobile phase was Methanol: Acetonitrile: Millipore Water (60:10:30) v/v ratio with the flow rate of 1ml/min at 292nm. The developed method was validated for the quantitative analysis of commercially available pure drugs. The instrument used was Shimadzu PDA-20AR, Lab solution software. The retention time for Lamivudine and Raltegravir were found to be 2.883 and 4.430 respectively. The linearity study of Lamivudine was performed by using concentration range of 1-20µg/ml and for Raltegravir concentration range was at 10-30µg/ml and correlation coefficient (r2) was found to be 0.999 and 0.9977 respectively. The developed method was validated in reference to ICH Guidelines. Specificity, Linearity, Accuracy, Robustness, Precision and Ruggedness obtained within the acceptance range.

Studying the cumulative vigor response index of morpho-physiological, quality, and yield-related traits of wheat cultivars using planting dates

Wheat (Triticum aestivum L) is one of the major staple food crops consumed globally. Nonetheless, the cultivation of wheat is influenced by various environmental factors, with the planting date being significantly impacted by the effects of climate change. Addressing these changes could involve evaluating wheat genotypes to identify appropriate planting dates. A phenotypic screening experiment was conducted in the field crop station of Agriculture College of Tikrit University to determine the suitable planting time for wheat cultivars under local environmental conditions during 2022-23. Several morpho-physiological, quality, and yield traits were measured. Factorial experiment using spilt plot through randomized completely block design (RCBD). was used with three replications. The five planting dates (5-10, 25-10, 15-11, 5-12, and 25-12) were considered as the main plot, and the eight wheat cultivars (Ipaa99, Al-Rasheed, Al-Baraka, Sham6, Tammuz2, Al-Hashimiya, Al-Noor, and Al-Adnanieh) as sub-main plot. Data were used to calculate the Individual, Cumulative, and Total Vigor Response Indices (IRI, CRI & TRI). Cultivars were classified into different categories using total cumulative early or late planting date vigor response index values (TRI-e) or (TRI-l) and standard deviation (SD). The (TRI-e) values ranged from 36.07 (sensitive) for the cultivar Al-Baraka to 39.13 (tolerant) for the cultivar Al-Hashimiya. However, the (TRI-l) values ranged from 36.59 (sensitive) for the cultivar Al-Noor to 39.52 (tolerant) for the cultivar Al-Hashimiya. The correlation coefficient (r2) between the (TRI-e) and cumulative very early/early planting date vigor response index was positively correlated (r2 = 0.70 for very early planting date (5-Oct) and r2 = 0.60 for early planting date (25-Oct). Furthermore, 76% of the total variation in the (TRI-l) was explained by the cumulative very late planting date vigor response index (CRI-vl) while just 49% of the total variation was explained by the cumulative late planting date vigor response index (CRI-l). Based on those results, wheat producers could select either tolerant cultivars for early planting or tolerant cultivars for late planting to maximize wheat production in their specific growing environments including planting dates.

In English

Currently, textile and food industries produce a significant volume of sewages containing azo dyes and other organic pollutants. These effluents are serious environmental threats, so new methods for their treatment and the degradation of azo dyes are attracting much attention. Composite materials based on TiO2 modified by noble metals and nanoceria show high activity in the photodegradation of organic contaminates and are proposed for hydrogen synthesis as well. To optimize the treatment of contaminants, different processes can combine including the strategies of adsorption, photoluminescence, photocatalysis, etc. The synthesized TiO2-based nanomaterials (sols, powders) will be exploited for bioremediation due to their small size and surface plasmon resonance from noble metals. Binary nanocomposites based on TiO2 were obtained by the chemical co-precipitation method from solutions of titanium tetraisopropoxide (TTIP) and inorganic salts of cerium, silver, and palladium. It has been stated that TiO2 is represented by anatase with primary particle size (CSR) from 8.5 to 16.8 nm, depending on the nature and concentration of the dopant. It is shown that Ag is reduced on the surface of anatase particles and blocks their growth, while Pd and Ce penetrate the titanium dioxide matrix in the form of small clusters with the deformation of the anatase crystal lattice. Nanocomposite particles formed loose and fragile aggregates, which spontaneously dispersed in solutions of dyes with the formation of colloid-stable sols, required the use of a centrifugal field for their sedimentation. Nanoparticles of TiO2&Pd were electronegative and others were electropositive according to the values 4.1÷9.6 of ZPC (zero point of charge). It was shown that the particles of all composites sorbed Methylene Blue (MB) without photocatalytic activity under the visible light to any dye. Moreover, anionic dyes such as Orange-G (Or-G) and Methyl Orange (MO) were excellently discolorated in the presence of TiO2&Pd system; cationic dyes of MB and Rhodamine B (RhB) discolorated too with the TiO2, TiO2&CeO2, and TiO2&Ag systems under UV light action. As such, photocatalysis tests showed that Orange-G’s and MO’s discoloration was higher for TiO2&Pd (2 wt. %) and TiO2 systems with the correlation coefficient R2 0.999.

Trace Level Quantification of Genotoxic Impurities through Atmospheric Pressure Chemical Ionization (APCI) Coupled with Triple Quardrapole Analyzer in Combination Diabetic Drug Product

N-Nitrosamine impurities have been found in several drug products, showing a concern for regulatory aspects. The study presents the development and validation of a sensitive LC-MS/MS method for detecting eight nitrosamines in vildagliptin and metformin drug products. The developed method was specific and linearity was ranged 3.53-55.92 ppb for NDMA, 3.18-50.37 ppb for NMBA, 0.97-15.41 ppb for NDEA, 0.98-15.52 ppb for NEIPA, 1.00-15.86 ppb for NDIPA, 0.98-15.46 ppb for NDBA, 0.98-15.58 ppb for NMPA and 0.7-15.32 ppb for NDPA with correlation coefficient (r) was more than 0.98 and square of correlation coefficient (r2) was found to be greater than 0.96. The LOQ were obtained in the range of 0.97-3.53 ppb with %RSD in the range of 2.5-10.9% for eight nitrosamines showed good sensitivity. Accuracy was found in the range of 98.63 ± 5.77-125.17 ± 3.68% at LOQ level, 74.77 ± 0.83-88.17 ± 0.15% at 50% level, 79.93 ± 8.47-89.00 ± 0.61% at 100% level and 74.67 ± 2.31-84.03 ± 0.93% at 150% level for the eight nitrosamine impurities that were well within acceptance criteria of not less than 70% and not more than 130%. The method validation results demonstrated that the method is precise, accurate and linear, can be applied to quantify the nitrosamines in vildagliptin and metformin drug products.

A Fuzzy-Based Analysis of Air Particle Pollution Data: An Index IMC for Magnetic Biomonitoring

Airborne magnetic particles may be harmful because of their composition, morphology, and association with potentially toxic elements that may be observed through relationships between magnetic parameters and pollution indices, such as the Tomlinson pollution load index (PLI). We present a fuzzy-based analysis of magnetic biomonitoring data from four Latin American cities, which allows us to construct a magnetic index of contamination (IMC). This IMC uses four magnetic parameters, i.e., magnetic susceptibility χ, saturation isothermal remanent magnetization SIRM, coercivity of remanence Hcr, and SIRM/χ, and proposes summarizing the information to assess an area based exclusively on magnetic parameters more easily. The fuzzy inference system membership functions are built from the standardization of the data to become independent of the values. The proposed IMC is calculated using the baseline values for each case study, similar to the PLI. The highest IMC values were obtained in sites close to industrial areas, and in contrast, the lowest ones were observed in residential areas far from avenues or highways. The linear regression model between modeled IMC and PLI data yielded robust correlations of R2 > 0.85. The IMC is proposed as a complementary tool for air particle pollution and is a cost-effective magnetic approach for monitoring areas.

Controllable functionalization of amino-functionalized graphene quantum dots as fluorescent probe for detection of Cu(Ⅱ) ions detection

Herein, the amino-functionalized graphene quantum dots (GQDs) can be applied for selective detection of Cu(Ⅱ) ions in solutions via photoluminescence (PL) measurement, the PL emission intensities of the GQDs gradually decreased with the increase of Cu2+ concentration (0–100 μm), the PL intensity (quenching effect) decreases almost linearly with increasing Cu2+ concentration. Its correlation coefficient R2 is 0.9529. Quenching of the PL emission by the presence of copper(Ⅱ) ions is attributed to the surface functional groups attached to the GQDs, which can interact with metal ions so that the electrons in the excited state of the GQDs are transferred into the unoccupied orbits of Cu(Ⅱ) ions. Corresponding physical mechanism is proposed to gain an understanding about how the presence of the Copper(Ⅱ) ions can affect the PL emission from GQDs.

Testing for concordance between predicted species richness, past prioritization, and marine protected area designations in the western Indian Ocean.

Scientific advances in environmental data coverage and machine learning algorithms have improved the ability to make large-scale predictions where data are missing. These advances allowed us to develop a spatially resolved proxy for predicting numbers of tropical nearshore marine taxa. A diverse marine environmental spatial database was used to model numbers of taxa from ∼1000 field sites, and the predictions were applied to all 7039 6.25-km2 reef cells in 9 ecoregions and 11 nations of the western Indian Ocean. Our proxy for total numbers of taxa was based on the positive correlation (r2=0.24) of numbers of taxa of hard corals and 5 highly diverse reef fish families. Environmental relationships indicated that the number of fish species was largely influenced by biomass, nearness to people, governance, connectivity, and productivity and that coral taxa were influenced mostly by physicochemical environmental variability. At spatial delineations of province, ecoregion, nation, and strength of spatial clustering, we compared areas of conservation priority based on our total species proxy with those identified in 3 previous priority-setting reports and with the protected area database. Our method identified 119 locations that fit 3 numbers of taxa (hard coral, fish, and their combination) and 4 spatial delineations (nation, ecoregion, province, and reef clustering) criteria. Previous publications on priority setting identified 91 priority locations of which 6 were identified by all reports. We identified 12 locations that fit our 12 criteria and corresponded with 3 previously identified locations, 65 that aligned with at least 1 past report, and 28 that were new locations. Only 34% of the 208 marine protected areas in this province overlapped with identified locations with high numbers of predicted taxa. Differences occurred because past priorities were frequently based on unquantified perceptions of remoteness and preselected priority taxa. Our environment-species proxy and modeling approach can be considered among other important criteria for making conservation decisions.

Performance of Mg/Al and Zn/Al Hydroxide Double Lamellar-Bentonite for Removal of Anionic Azo Dye from Aqueous Solution

This paper presents the preparation and characterization of bentonite coated with hydroxide double lamellar Mg/Al-bentonite and Zn/Al-bentonite as a potential adsorbent material. The coating process involved co-precipitation of mixed metal nitrate solution (Mg-Al) or (Zn-Al), followed by immersion of bentonite (B-Na+) dispersion. The structures and morphologies of the coated bentonites were characterized using XRD, FTIR, BET, and SEM analysis. The results of the BET analysis indicate that Mg/Al-bentonite and Zn/Al-bentonite have larger surface areas and pore volumes compared to bentonite alone. Specifically, the surface area of Mg/Al-bentonite is 209.25 m2/g with a pore volume of 0.423 cm3/g, while Zn/Al-bentonite has a surface area of 175.95 m2/g and a pore volume of 0.313 cm3/g. In contrast, the surface area and pore volume of bentonite alone are 110.43 m2/g and 0.132 cm3/g, respectively. The Mg/Al-bentonite reaches 85% uptake within 3 h (equivalent to 724.20 mg/g at 25 °C and pH 7), achieving rapid equilibrium. In contrast, the Zn/Al-bentonite achieves a maximum adsorption of 74% within 5 h under identical pH and temperature conditions, corresponding to 650.34 mg/g. The error function values, including the correlation coefficient R2, chi-square test χ2, and residual sum of squares RSS, were calculated to evaluate both kinetic and isotherm models. The kinetic adsorption data agreed well with a pseudo-second-order model. The adsorption process followed the Sips isotherm model, and the monolayer adsorption capacity of Mg/Al-bent and Zn/Al-bent composites was 872.41 (R2 = 0.974) and 678.45 mg/g (R2 = 0.983), respectively. The thermodynamic analysis of the adsorption process revealed that it occurred spontaneously with an endothermic characteristic. The parameters ΔS, ΔH, and ΔG were used to determine this.

Preparation and Characterization of Periwinkle Shell Based Chitosan-Kenaf fibre Copolymer and its Derivatives for Selective Binding of Cu (II) and Zn (II) ions from Electroplating Effluent

The levels of heavy metal in industrial waste streams, if not remediated before discharge to eco-system, could pose a threat to flora and fauna. The objective of this study was to evaluate the suitability of a novel adsorbent materials from periwinkle shell-based chitosan-kenaf fibre copolymer and its ethylene glycol diglycidyl ether crosslinked derivatives for the selective removal of Cu2+and Zn2+ from electroplating effluent using batch experimental protocol. Results showed that the crosslinked copolymers were more effective for the sorption of these metal ions from effluent solution with percentage sorption range of 76-87% and 84-93% for Cu2+ and Zn2+ respectively. Kinetic data depicts pseudo-second-order model fits for the adsorbent materials and confirm chemosorption as the mechanism for metal ion complexation with adsorbent surfaces. Based on R2 correlation values, sorption of the metal ions by the pristine adsorbent materials fit well with Langmuir isotherm model, in contrast, those for crosslinked adsorbent materials were best described by Freundlich isotherm model. The results present the adsorbent materials with auspicious potentials, especially the crosslinked derivatives, and stand them in exceptional good stead as suitable bioresource materials for column packing in waste abatement equipment for remediation of these metal ions from electroplating effluent solution.

Strategic Approaches to Elevate Quality and Sustainability in Drug Development: Comprehensive Pretomanid (PA-824) Chemical Stability study using QbD and Green Chemistry Principles

Regulatory agencies prioritize the safety and effectiveness of pharmaceuticals. The focus here is to highlight the crucial need for control and oversight of Pretomanid (PMD), a synthetic nitroimidazooxazine utilized in treating multidrug-resistant tuberculosis. This study aims to develop a simple, sensitive, LCMS compatible, and environmentally friendly HPLC Stability Indicating Analytical Method following ICH Q1A (R2) and Q3 guidelines. The approach involves the application of Quality by Design (QbD) principles and adherence to green chemistry practices. A comprehensive risk assessment was conducted using cause-and-effect diagram and risk assessment tool. The QbD approach includes screening and optimization phases. Plackett-Burman design was performed for screening selected 11 primary parameters. Central composite design was used for optimisation of selected critical variables based on critical method parameters (CMPs) and Critical method attributes (CMAs). The method was optimised using Contour plot, 3D plot, ANOVA, Desirability value and Design space. Superior component separation was achieved through an isocratic elution using Kromasil C18 (250 × 4.6 mm; 5 μm) column, mobile phase composition of (50:50; v/v) ACN: 10 mM Ammonium Acetate pH 4.5 adjusted with Glacial Acetic acid and a flow rate of 0.9 mL/min having runtime of 30 mins. The stress studies revealed that the drug is highly sensitive to hydrolysis, oxidation, and photolytic conditions. Experimentation results demonstrated that acetonitrile served as the most suitable diluent. Additionally, the correlation coefficient (r2) exceeded 0.996, while the RSD values (n = 6) varied from 0.76 % to 1.93 % across the LOQ–150 % range. Specificity studies revealed no interference between peaks from impurities and known active analytes. Through stress studies, major degradation products were identified. The method's eco-friendliness was assessed using ComplexGAPI and AGREE, confirming its environmentally friendly nature.

An Ecologically Sound RP-HPLC Method for Estimating Citicoline using AQbD with Degradation Studies

Background: Citicoline is widely used for neurological complications. This research work was carried out using reverse-phase high-performance liquid chromatography (RP-HPLC) employing green analytical chemistry principles. Materials and Methods: The method was completely developed with ethanol:water (20:80 v/v) as the mobile phase, 0.1% orthophosphoric acid was used to adjust the pH to 3.5, and an Inertsil ODS 3 (250 x 4.6 mm, 5 μm) column as the stationary phase, the flow rate was set at 1-mL/min. The column temperature was maintained at 35ºC, and an injection volume of 10 μL was used for detection at a wavelength of 250 nm. Results and Discussion: The peak was eluted with isocratic elution with a retention time of 3.996 minutes. The linear concentration range for this method was 10 to 50 μg/mL, and the correlation coefficient R2 was 0.999. The forced degradation studies have shown that 3.5% w/w degraded in acid, 1.35% w/w in alkali, and 0.2% w/w in oxidation. The products formed during degradation were characterized using gas chromatography-mass spectrometry (GC-MS). Conclusion: The International Council for Harmonisation (ICH) guidelines were followed in the development and validation of the method optimized by the use of central composite design, and the method’s greenness was assessed through the use of AGREE, AES, GAPI, and AMGS, among other green assessment tools.

Novel HPLC Method Validation for Quantification of Dapagliflozin in Bulk Drug and Formulation

A new, steady, easy-to-use, and straightforward high-performance liquid chromatography (HPCL) technique was also tested to determine dapagliflozin concentration in medicinal and API forms. For the chromatographic extraction study, a Phenomenex Gemini-NX C18 Column was used. This column was filled with a mobile phase that was 70:30 v/v methanol to sodium 1-octanesulphonate. This experiment used a 20-liter syringe amount and a flow rate of 1-mL per minute. A bunch of factors were used to make sure the new method worked. The drug dapagliflozin was released at a wavelength of 203 nm after 6.5 ± 0.5 minutes. A correlation coefficient (r2) of 0.9994 between 15 to 55 μg/mL shows that the planned process is linear. The theoretical plates and peak tailing were found to be 4129 and 1.03, respectively. In terms of healing, the percent accuracy that was reached was within the acceptable range of 98 to 100. New suggestions say that the regular testing of dapagliflozin in large amounts must be done using a proven method that is unique and easy to repeat. Forced breakdown tests with drugs were used to show that the proposed method is stable. The product was tested according to the steps laid out in the ICH Q2R1 Guidelines. The test looked at many chemical processes, such as oxidation, photolysis, heat stress, and acids and alkaline hydrolysis.

Development and Validation of UPLC Tandem Mass Spectrometry Assay for Ceftibuten and Sulbactam in Human Plasma.

A sensitive and rapid ultra-performance liquid chromatography coupled with -tandem mass spectrometry (UPLC-MS/MS) method was developed and validated to determine ceftibuten (CTB) and sulbactam (SUL) in human plasma. An ACQUITY UPLC HSS T3 C18 (2.1 × 100 mm), 1.8 μm column with gradient elution of water (0.1% formic acid) and acetonitrile was used for separation at a flow rate of 0.2mL/min. This method involves a simple sample preparation with acetonitrile. The calibration curves of CTB and SUL in plasma showed good linearity over the concentration range of 0.50-25 μg/mL and with a correlation coefficient (r2) >0.99. This method was validated in terms of selectivity, linearity, precision, accuracy and stability. High precision was obtained with coefficients of variation <15%. Excellent recovery in the range of 90-104% was achieved for CTB and SUL was 86-110%. The method has the potential utility to support pharmacometric modeling in clinical practice and biopharmaceutic studies.

Abstract 6878: Images to insights: Transforming TME characterization into actionable spatial metrics with AI-powered multiplex immunofluorescence

Abstract Background: Precise and actionable assessment of cellular mechanisms and interactions within the tumor microenvironment (TME) is critical for advancing clinical trials, preclinical research, and informed decision-making in immuno-oncology. Multiplex immunofluorescence (mIF) assays have emerged as pivotal tools that can enable comprehensive visualization of protein expression and cellular interactions within the TME. Integration of advanced digital analysis tools and mIF assays, can revolutionize our ability to interpret the TME and usher these assays into high-throughput clinical settings. Methods: In this study, we processed tumor samples utilizing Akoya's PhenoCode Signature Immune Profile Human Protein Panel, which includes immune profiling markers CD8, CD68, CD3, CD20 and tumor marker PanCK. Following the Akoya Opal staining protocol, formalin-fixed paraffin-embedded (FFPE) samples were stained and scanned using the PhenoImager HT scanner. The images were then analyzed using Reveal’s AI-mIF digital assay on the ImageDx platform to generate contextual and spatial biomarker data. A deep learning-based cell segmentation model based on MaskRCNN architecture, signal preprocessing for noise and autofluorescence, and dynamic intensity normalization were used. Phenotype classification was precisely executed using dynamic thresholding or neural network classifiers as needed. Results were validated by establishing concordance with pathologist readouts. Results: The application of our methodology yielded highly concordant results when compared with manual counts, demonstrating the efficacy of our approach. The R2 correlation coefficient across all markers passed the minimum criteria of R2&amp;gt;0.85 with regression slopes between 0.7-1.3, indicating a robust alignment with traditional pathologist assessments. We showcase stratifying samples Desert, Excluded, or Inflamed phenotypes and other quantitative and spatial results such as interactions of immune cells and tumor within 30-um neighborhoods, and their dispersion and arrangement in the TME. Conclusion: The integration of Reveal Biosciences' ImageDx platform and RevealAI-mIF provides an ecosystem to host, visualize, and analyze Akoya’s immune profiling images. This collaborative effort has enabled the transformation of complex multiplex imaging data into distinct spatial phenomic endpoints, thereby introducing the potential to significantly advance biomarker discovery and contribution to translational and clinical research in the field of immuno-oncology. Citation Format: Misagh Naderi, Lazar Krstic, Djordje Cikic, Kristen Ruf, Igor Mihajlovic, Sinisa Todorovic, Claire Weston. Images to insights: Transforming TME characterization into actionable spatial metrics with AI-powered multiplex immunofluorescence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6878.

First Specific Detection of Mammalian Orthoreovirus from Goats Using TaqMan Real-Time RT-PCR Technology.

Mammalian orthoreovirus (MRV) infections are ubiquitous in multiple mammalian species including humans, and mainly causes gastroenteritis and respiratory disease. In this study, we developed a rapid and sensitive TaqMan qRT-PCR method for MRV detection based on the primers and probe designed within the conserved L1 gene. The qRT-PCR assay was evaluated for its sensitivity, specificity, efficiency and reproducibility. It was found that the detection sensitivity was equivalent to 10 DNA copies/μL, and the standard curves had a linear correlation of R2 = 0.998 with an amplification efficiency of 99.6%. The inter- and intra-assay coefficients of variation (CV%) were in the range of 0.29% to 2.16% and 1.60% to 3.60%, respectively. The primer sets specifically amplified their respective MRV segments and had the highest detection sensitivities of 100.25 TCID50/mL with amplification efficiencies of 99.5% (R2 = 0.999). qRT-PCR was used for MRV detection from samples of sheep, goats, and calves from four regions in China, and the overall MRV prevalence was 8.2% (35/429), whereas 17/429 (4.0%) were detected by RT-PCR and 14/429 (3.3%) by virus isolation. The qRT-PCR assay showed significantly higher sensitivity than RT-PCR and virus isolation. Results from an epidemiological survey indicated that the positive rate of MRV in rectal swabs from sheep and goats tested in Shaanxi, Jiangsu, and Xinjiang were 9/80 (11.3%), 12/93 (12.9%) and 14/128 (10.9%), respectively. In goats and sheep, MRV prevalence was obviously associated with season and age, with a high positive rate of more than 8% during September to April and approximately 13% in small ruminant animals under two months of age. This is the first instance of MRV infection in sheep and goats in China, thus broadening our knowledge of MRV hosts. Consequently, primer optimization for qRT-PCR should not only prioritize amplification efficiency and specificity, but also sensitivity. This assay will contribute to more accurate and rapid MRV monitoring by epidemiological investigation, viral load, and vaccination efficacy.

Abstract 335: BD®OMICS-Guard Sample Preservation Buffer preserves mRNA and cell surface epitopes for single-cell sequencing applications

Abstract Multiomics has emerged as a powerful tool to derive insights on the transcriptomic and proteomic data of human cancer samples and preclinical models of blood and tumor malignancies. Due to the sensitivity of sequencing techniques, sample quality and stability can potentially interfere with results. BD® OMICS-Guard Sample Preservation Buffer (SPB), a novel preservation solution, allows cells and bulk tissues to be stored for up to 72h at 4 °C, while reliably capturing the true biology of cells. This reagent gently preserves mRNA and protein integrity without the traditional cross-linking and harsh fixatives, increasing flexibility and expanding options for study designs. Here, we showcase the preservation properties ofthis reagent for multiomic readouts in human cells and mouse tissue across three different timepoints.BD® OMICS-Guard SPB’s preservation of mRNA from human PBMC and bulk mouse spleen tissues wasevidenced by WTA for differentially expressed genes in samples preserved for 24, 48 and 72h comparedto fresh/unpreserved controls. Protein epitope preservation was demonstrated via CITE-Seq assay withthe BD® AbSeq Human Immune Discovery Panel (human PBMC) or a 30-plex panel of anti-mouse BD®AbSeq Antibody-Conjugated Oligonucleotides (mouse splenocytes) and confirmed with multicolor flowcytometry. For both sample types, high correlation R2 values (&amp;gt;0.87) were calculated for the preservation time points versus fresh controls. Concomitant flow cytometry analysis of major cell types and surface protein expression was consistent with sequencing data for all samples. Specificity and sensitivity of major cell type markers and cell surface proteins across cell types and time is visualized in the BD®AbSeq Antibody-generated heatmap for each sample. Additionally, BD® OMICS-Guard SPB maintains mRNA integrity in targeted gene assays and VDJ full-length assays, suggesting robust preservation of select transcripts readily interrogated in single-cell profiling. Batch effect across time and technical replicates for both sample types was limited with the BD Rhapsody™ HT Xpress System and delineated by donor/sample specific tSNEs across time. We demonstrate that the BD® OMICS-Guard Sample Preservation Buffer effectively preserves both mRNA and surface proteins for single-cell experiments. BD® OMICS-Guard SPB allows the convenient preservation of samples such as tumor biopsies to be collected and stored for up to 72h at 4 °C while maintaining mRNA and protein integrity, allowing the flexibility for precious samples to be potentially shipped to a central sequencing site to ensure standardized sequencing workflows. For Research Use Only. Not for use in diagnostic or therapeutic procedures.BD and BD Rhapsody are trademarks of Becton, Dickinson and Company or its affiliates. © 2023 BD. All rights reserved. Citation Format: Samuel X. Shi, Cynthia Sakofsky, Zhiqi Zhang, Samatha Vadrevu, Xiaoshan Shi, Hye-Won Song, Aruna Ayer. BD®OMICS-Guard Sample Preservation Buffer preserves mRNA and cell surface epitopes for single-cell sequencing applications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 335.

Length-Weight Relationship and Condition Factor of a Commercial Food Fish Labeo rohita (Hamilton, 1822) from Ghaghara River, Uttar Pradesh, India

The present study provides first-hand information about the length-weight relation of Labeo rohita (Hamilton) from Ghaghara River, Uttar Pradesh. We collected 107 specimens and took length and weight for better conservation techniques and proper management for rohu. A total of 107 specimens were examined for the study, which has ranging from 25.2-56.13 cm TL (Total length) and 183.3-2409.78 gm (weight) were used for the study of LWRs and condition factors. Our study represented that L. rohita exhibited a negative allometric growth (b &lt; 3) indicating that the fish grows faster in length compared to their weight. The values of the correlation coefficient (r2) and condition factor (K) are 0.8601 and 1.27 respectively. This research will be useful for fishery research, stock assessment, conservation, and estimation of the fish condition of this Himalayan River.

Revisiting methotrexate and phototrexate Zinc15 library-based derivatives using deep learning in-silico drug design approach.

Introduction: Cancer is the second most prevalent cause of mortality in the world, despite the availability of several medications for cancer treatment. Therefore, the cancer research community emphasized on computational techniques to speed up the discovery of novel anticancer drugs. Methods: In the current study, QSAR-based virtual screening was performed on the Zinc15 compound library (271 derivatives of methotrexate (MTX) and phototrexate (PTX)) to predict their inhibitory activity against dihydrofolate reductase (DHFR), a potential anticancer drug target. The deep learning-based ADMET parameters were employed to generate a 2D QSAR model using the multiple linear regression (MPL) methods with Leave-one-out cross-validated (LOO-CV) Q2 and correlation coefficient R2 values as high as 0.77 and 0.81, respectively. Results: From the QSAR model and virtual screening analysis, the top hits (09, 27, 41, 68, 74, 85, 99, 180) exhibited pIC50 ranging from 5.85 to 7.20 with a minimum binding score of -11.6 to -11.0 kcal/mol and were subjected to further investigation. The ADMET attributes using the message-passing neural network (MPNN) model demonstrated the potential of selected hits as an oral medication based on lipophilic profile Log P (0.19-2.69) and bioavailability (76.30% to 78.46%). The clinical toxicity score was 31.24% to 35.30%, with the least toxicity score (8.30%) observed with compound 180. The DFT calculations were carried out to determine the stability, physicochemical parameters and chemical reactivity of selected compounds. The docking results were further validated by 100ns molecular dynamic simulation analysis. Conclusion: The promising lead compounds found endorsed compared to standard reference drugs MTX and PTX that are best for anticancer activity and can lead to novel therapies after experimental validations. Furthermore, it is suggested to unveil the inhibitory potential of identified hits via in-vitro and in-vivo approaches.