Quality Control Standards Research Articles

Metabolic Pathway Engineering Improves Dendrobine Production in Dendrobium catenatum.

The sesquiterpene alkaloid dendrobine, widely recognized as the main active compound and a quality control standard of medicinal orchids in the Chinese Pharmacopoeia, demonstrates diverse biological functions. In this study, we engineered Dendrobium catenatum as a chassis plant for the production of dendrobine through the screening and pyramiding of key biosynthesis genes. Initially, previously predicted upstream key genes in the methyl-D-erythritol 4-phosphate (MEP) pathway for dendrobine synthesis, including 4-(Cytidine 5'-Diphospho)-2-C-Methyl-d-Erythritol Kinase (CMK), 1-Deoxy-d-Xylulose 5-Phosphate Reductoisomerase (DXR), 2-C-Methyl-d-Erythritol 4-Phosphate Cytidylyltransferase (MCT), and Strictosidine Synthase 1 (STR1), and a few downstream post-modification genes, including Cytochrome P450 94C1 (CYP94C1), Branched-Chain-Amino-Acid Aminotransferase 2 (BCAT2), and Methyltransferase-like Protein 23 (METTL23), were chosen due to their deduced roles in enhancing dendrobine production. The seven genes (SG) were then stacked and transiently expressed in the leaves of D. catenatum, resulting in a dendrobine yield that was two-fold higher compared to that of the empty vector control (EV). Further, RNA-seq analysis identified Copper Methylamine Oxidase (CMEAO) as a strong candidate with predicted functions in the post-modification processes of alkaloid biosynthesis. Overexpression of CMEAO increased dendrobine content by two-fold. Additionally, co-expression analysis of the differentially expressed genes (DEGs) by weighted gene co-expression network analysis (WGCNA) retrieved one regulatory transcription factor gene MYB61. Overexpression of MYB61 increased dendrobine levels by more than two-fold in D. catenatum. In short, this work provides an efficient strategy and prospective candidates for the genetic engineering of D. catenatum to produce dendrobine, thereby improving its medicinal value.

Research on the quality markers of antioxidant activity of Kai-Xin-San based on the spectrum-effect relationship.

Background: Kai-Xin-San (KXS) is one of the classic famous traditional Chinese medicine prescriptions for amnesia, which has been applied for thousands of years. Modern pharmacological research has found that KXS has significant therapeutic efficacy on nervous system diseases, which is related to its antioxidant activity. However, the antioxidant material basis and quality markers (Q-makers) of KXS have not been studied. Objective: The objective of this study is to explore the Q-makers of antioxidant activity of KXS based on spectrum-effect relationship. Methods: Specifically, the metabolites in KXS extracts were identified by UPLC-Q-Exactive Orbitrap MS/MS. The fingerprint profile of KXS extracts were established by high-performance liquid chromatography (HPLC) and seven common peaks were identified. Meanwhile, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) test was used to evaluate the free radical scavenging ability of KXS. The spectrum-effect relationship between its HPLC fingerprint and DPPH free radical scavenging activity was preliminarily examined by the Pearson correlation analysis, grey relation analysis (GRA), and orthogonal partial least squares discrimination analysis (OPLS-DA). Further, the antioxidant effect of KXS and its Q-makers were validated through human neuroblastoma (SH-SY5Y) cells experiment. Results: The results showed that 103 metabolites were identified from KXS, and the similarity values between HPLC fingerprint of twelve batches of KXS were greater than 0.900. At the same time, the results of Pearson correlation analysis showed that the peaks 8, 1, 14, 17, 18, 24, 16, 21, 15, 13, 6, 5, and 3 from KXS were positively correlated with the scavenging activity values of DPPH. Combined with the results of GRA and OPLS-DA, peaks 1, 3, 5 (Sibiricose A6), 6, 13 (Ginsenoside Rg1), 15, and 24 in the fingerprints were screen out as the potential Q-makers of KXS for antioxidant effect. Besides, the results of CCK-8 assay showed that KXS and its Q-makers remarkably reduced the oxidative damage of SH-SY5Y cells caused by H2O2. However, the antioxidant activity of KXS was decreased significantly after Q-makers were knocked out. Conclusion: In conclusion, the metabolites in KXS were successfully identified by UPLC-Q-Exactive Orbitrap MS/MS, and the Q-makers of KXS for antioxidant effect was analyzed based on the spectrum-effect relationship. These results are beneficial to clarify the antioxidant material basis of KXS and provide the quality control standards for new KXS products development.

Screening for Q-markers of Antiviral Granules based on neuraminidase inhibitors and Effect-constituent index

BackgroundAntiviral granules (AG) is a classic traditional Chinese medicine commonly employed for influenza treatment. However, there is an urgent need to improve its quality control standards. PurposeThe quality markers (Q-markers), guided by the Effect-constituent Index (ECI), were screened through in vitro and in vivo component identification, molecular docking, and neuraminidase (NA) inhibition experiments. MethodsThe chemical components in AG underwent analysis using UPLC-Q-Orbitrap HRMS and GC–MS, and serum pharmacochemistry was employed to analyze the components absorbed into the bloodstream. Molecular docking was used to evaluate the affinity of the prototype components to NA, and the inhibitory activity of the candidate components was later confirmed through NA inhibitor experiments. Following this, a UHPLC-MS/MS method was developed to screen and quantify the Q-markers in 20 sample batches. Lastly, the ECI was formulated using a multi-index comprehensive evaluation method that combines composition, content, and activity. ResultsA total of 246 compounds were identified in AG, and 89 prototype components and 19 metabolic components were detected in the serum of the rats. Molecular docking revealed that the binding energy of 35 candidate active ingredients was better than that of the original ligand and the target protein. Among these candidate active ingredients, 12 candidate Q-markers demonstrated significant inhibitory effects in NA inhibition experiments, with IC50 values ranging from 0.954 to 3.408 mM. A UHPLC-MS/MS content determination method was established for the quantification of 11 Q-markers, including Forsythiaside A, Pogostone, Verbascoside, Mangiferin, 5-methoxy-8-hydroxypsoralen, Curcumenol, Luteolin-7-glucuronide, Kaempferol-3-O-rutinoside, Baicalin, Rutin, and Bergaptol, in 20 batches of samples. The ECI was calculated based on activity and content of these Q-markers. Pearson's correlation analysis of the NA inhibitory effect of the samples and the ECI resulted in an R value of 0.859, indicating a good correlation. Furthermore, this correlation coefficient surpassed those observed between the content of each Q-marker and the NA inhibitory effect. ConclusionThe Q-markers for treating influenza with AG have been successfully screened, and the ECI of AG has been constructed. These findings will provide beneficial strategies for improving the quality control level of AG.

NIR-hyperspectral imaging and machine learning for non-invasive chemotype classification in Cannabis sativa L

The current public acceptance rate towards medical cannabis feasibility has led to a worldwide increase in this plant species production. Nevertheless, the currently transforming legal framework does not prevent the originally unlawful knowledge around cannabis breeding, which lacks quality control regulations or standards for correct manufacturing processes, a fact that could subsequently lead to uncontrolled and even harmful crop products. In this line, the objective of this work was to develop a non-invasive methodology for cannabis chemotype classification in different cultivars during the plant cultivation process, in order to keep undoubtful production control over cannabis crops. Hence, hyperspectral imaging (HSI), coupled with various multivariate data analysis approaches, such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), enabled the non-invasive in-situ analysis of the plants. Hence, two PLS-DA classification models were trained with the plant spectral data for three chemotypes, based on the cannabinoid content of the plant inflorescences, with the difference between both approaches being the regard of the stem part of the plant as a bias. Thus, obtained sensitivity and specificity values in the inflorescences were 0.845/0.845 for Chemotype I, 0.954/0.920 for Chemotype II, and 0.888/0.925 for Chemotype III. At last, a hierarchical PLS-DA, which considered the stem as a bias, presented an overall 94.7 % trueness in the external validation of 57 different plant individuals, divided as 92.3 % trueness for chemotype I, 100.0 % trueness for chemotype II and 88.9 % trueness for chemotype III. Based on these results, the proof of concept for comprehensive agricultural control of cannabis crops through a non-invasive analytical technique was demonstrated, a previously unproven fact. Therefore, this work could further pave the way for non-invasive technology development for horticultural quality control in medical cannabis productions, as this emerging industry will require strict control over the cannabis chemotypes, with the strong advantage of avoiding destructive and time-consuming analytical techniques such as chromatography.

Validation and Determination of Mercury in Yemeni Common Canned Tuna Using Direct Mercury Analyzer

The presence of Hg in canned tuna flesh is a major source of customer concern. Thus, the primary goal of this investigation was to evaluate total mercury levels in Yemeni canned tuna. Thirty-one samples of seven popular locally canned tuna brands were acquired from local stores in Sana'a. A Direct Mercury Analyzer was used to determine the mercury content. The linearity of the method was tested by assessing a series of mercury standards in triplicates at concentrations ranging from 20 to 500 ppb. The correlation coefficient for this method was 0.9993. The repeatability of the procedure was measured as intra-day variation and given as the relative standard deviation, which did not exceed 4.45%. This indicated the excellent precision of the method. Mercury recoveries from spiked actual canned tuna samples were between 96.61% and 100.49%, demonstrating excellent accuracy and an acceptable procedure for mercury measurement. The limits of detection and quantitation were 1.62 ppb and 5.4 ppb, respectively. The mercury content in 31 canned tuna samples was less than the 500 ppb standard imposed by the Yemeni Organization for Standards Metrology and Quality Control (YSMO).

Evaluating Pfeiffer Circular Chromatography as a Qualitative Methodology of Viscum album Homeopathic Mother Tinctures: A Multicentric Collaborative Project Between Brazil and Switzerland

Viscum album (VA) is a peculiar semiparasitic plant with important role in cancer therapy. The differences in phytochemical composition as well as the influence of host tree in the metabolome of VA subspecies are already described1. Our goal is to use the Pfeiffer Circular Chromatography (PCC) as a qualitative methodology to differentiate V. album mother tinctures (VAMT). The assays were conducted in two countries: Brazil (São Paulo and Rio de Janeiro) and Switzerland (Arlesheim). Three VA subspecies (abietis, album, austriacum) and five host trees (Malus domestica, Quercus sp., Ulmus carpinifolia, Pinus sylvestris, Abies alba) were harvested in Switzerland in summer and winter. The VAMTs were prepared following the Brazilian Homeopathic Pharmacopoeia and the PCC experiments were performed as previously published2. The preliminary analysis and the reproducibility of the three independent assays performed in each experimental set are being evaluated by Image J software. The qualitative standard features will be used to the VAMT differentiation regarding the following PCC characteristics: inner and outer-zones, coloration, size, the presence of radial lines, among others. Besides, the High-Performance Thin Layer Chromatography (HPTLC) was performed as standard quality control methodology using the CAMAG instruments to compare the phytochemical profile of V. album mother tinctures described in the French and German Homeopathic Pharmacopoeias. The samples have been characterized by the compounds retention factor (Rf) compared to the chemical standards (chlorogenic acid, caffeic acid, hyperoside and rutin). They present similar chemical markers when both methodologies were compared but with different intensities between season and host trees. Based on the hypothesis that the PCC patterns could be correlated with the chemical composition of VAMT, this multicentric research project will be useful to propose a low-cost and an easy methodology focused on the quality control parameters of these herbal raw materials.

Searching for Optimal Measurement Parameters by Thermogravimetry for Determining the Degree of Modification of Thermally Modified Wood

When wood is thermally modified, several chemical reactions take place that change the chemical and physical properties of the wood. These changes correlate with the degree of modification, which is mostly a function of the temperature and duration of modification, and consequently with the mass loss during this process. There is a lack of standardised quality control to verify the degree of heat treatment of wood and thus its quality. One of the possible methods to check the degree of thermal modification of a particular type of wood is thermogravimetry (TG). It is based on the assumption that processes that did not take place during thermal modification continue when the TG experiment is carried out. In this method, calibration curves have to be established based on TG measurements of standard samples that have been thermally modified at different temperatures and whose mass loss during modification is known. The calibration curves show the mass loss during the TG measurement as a function of the mass loss during the previous thermal modification. The course of thermal decomposition during the TG measurements is influenced by many parameters, such as the mass of the sample, the heating rate, the atmosphere in which the measurement takes place, and the shape of the crucible in which the sample is placed. In this paper, the influence of these parameters on the calibration curves was investigated. We have focused on oak wood. The best parameters result in a calibration curve with the largest correlation coefficient R2 and the highest slope of the line k. On this basis, we can determine the mass loss during the thermal modification of unknown samples of the same wood species under the same measurement conditions.

Use of continuous genotypes for genomic prediction in sugarcane.

Genomic selection in sugarcane faces challenges due to limited genomic tools and high genomic complexity, particularly because of its high and variable ploidy. The classification of genotypes for single nucleotide polymorphisms (SNPs) becomes difficult due to the wide range of possible allele dosages. Previous genomic studies in sugarcane used pseudo-diploid genotyping, grouping all heterozygotes into a single class. In this study, we investigate the use of continuous genotypes as a proxy for allele-dosage in genomic prediction models. The hypothesis is that continuous genotypes could better reflect allele dosage at SNPs linked to mutations affecting target traits, resulting in phenotypic variation. The dataset included genotypes of 1318 clones at 58K SNP markers, with about 26K markers filtered using standard quality controls. Predictions for tonnes of cane per hectare (TCH), commercial cane sugar (CCS), and fiber content (Fiber) were made using parametric, non-parametric, and Bayesian methods. Continuous genotypes increased accuracy by 5%-7% for CCS and Fiber. The pseudo-diploid parametrization performed better for TCH. Reproducing kernel Hilbert spaces model with Gaussian kernel and AK4 (arc-cosine kernel with hidden layer 4) kernel outperformed other methods for TCH and CCS, suggesting that non-additive effects might influence these traits. The prevalence of low-dosage markers in the study may have limited the benefits of approximating allele-dosage information with continuous genotypes in genomic prediction models. Continuous genotypes simplify genomic prediction in polyploid crops, allowing additional markers to be used without adhering to pseudo-diploid inheritance. The approach can particularly benefit high ploidy species or emerging crops with unknown ploidy.

Quantitative Amyloid‐PET in Real‐World Practice: Lessons from the Imaging Dementia—Evidence for Amyloid Scanning (IDEAS) study

AbstractBackgroundAmyloid‐PET quantification was developed in research settings using highly selected samples, harmonized acquisition protocols, and MRI‐based preprocessing. We aimed to analyze scans from IDEAS, a large‐scale, real‐world study of amyloid‐PET. Major challenges lie in scans being acquired on various PET/PET‐CT/PET‐MR scanners without standardization of acquisition or reconstruction protocols, and without MRI to preprocess data.Method18,295 Medicare beneficiaries with MCI or dementia underwent PET with [18F]florbetapir, [18F]florbetaben, or [18F]flutemetamol at 343 facilities. Sites shared 10,700 raw scans with their visual interpretation (positive or negative) by local radiologists/nuclear medicine physicians. Scans were processed using a PET‐only pipeline, which derives Centiloid based on a cortex‐to‐cerebellum tissue ratio in template space. Processed scans underwent standard visual quality control by 2 raters (Fig 1a‐b), and flagged scans were reviewed in a consensus meeting to discuss troubleshooting options. Injected dose and time between tracer injection and PET acquisition start were obtained from DICOM file header.Result9,963 scans (93%) were successfully pre‐processed with default parameters (Fig 1a) and 403 additional scans (4%) were included after manual intervention (Fig 1c). 339 scans (3%) were excluded because of technical artifacts or major bilateral lesions (Fig 1c). The final sample (n = 10,361) is described in Table 1, with example scans in Fig 1d. Centiloids showed the expected bimodal distribution (Fig 2a) and differentiated visually positive from negative scans (median = ‐2 versus 74 CL; area under the curve = 0.913, Fig 2b). PET acquisition generally followed tracer‐specific FDA recommendations for injected dose and acquisition time, but major variability was observed (Fig 2c). Centiloid values were independent from injected dose but were higher in patients scanned at later time windows for [18F]flutemetamol, resulting from known slow tracer kinetics (Fig 2c).ConclusionMRI‐free, amyloid‐PET quantification in a real‐world setting was feasible, with results conforming to expected distribution of Centiloid values and concordant with visual reads. However, greater standardization of PET acquisition than is currently required by FDA labels will be needed to reliably quantify amyloid PET in clinical practice (e.g., to monitor response to amyloid lowering therapy). The IDEAS dataset will be made available via the Global Alzheimer’s Association Interactive Network in 2023.

Determination and Method Optimization of Lianjie Liyan Compound Mixture

Euchresta japonica, Platycodon grandiflorum, Belamcanda chinensis, and Adenophora tetraphylla are the four medicinal substances that make up the Lianjie Liyan compound mixture, a type of compound preparation for treating throat diseases such as pharyngitis and laryngeal carcinoma. The preparation has been used clinically for a long time and has been proven that it has curative properties in terms of alleviating pain and other symptoms. However, no quality control research on this preparation has been done yet. The pharmacological components in the main quality control standards, including Oxymatrine, matrine, N-methylcytisine, platycoside-D in Platycodon grandiflorum, and irisaxanthin in Shegan, were analyzed to further standardize the clinically rational use of this preparation and prevent the occurrence of adverse reactions and side effects. In this study, the five active components in the mixture were determined concurrently for the first time with the help of Box-Behnken design and response surface approach. The properties of established model showed excellent resolution and sensitivity, good reproducibility and speed, and accurate prediction thanks to the optimization of HPLC measurement conditions. The established approach was precise and trustworthy, and it offered helpful support and references for the construction of the Lianjie Liyan compound mixture as well as for the quality control.

Genome‐wide identification of modifiers of age‐at‐onset for Alzheimer’s disease in African American populations

AbstractBackgroundSeveral studies have attempted to identify genes for age‐at‐onset (AAO) of Alzheimer’s disease (AD) through genome‐wide linkage and genome‐wide association studies (GWAS). While these efforts have thus far focused predominantly on non‐Hispanic white (NHW) populations, we present here the first GWAS to identify genetic contributors to AAO in African Americans (AA) using AA genotype and phenotype data from the Alzheimer’s Disease Genetics Consortium (ADGC).MethodWe combined data from 13 AA prospective and case‐control ADGC datasets, excluding family‐based datasets. Genotype data underwent standard quality control and were imputed to the TOPMed R5 reference panel using the Michigan Imputation Server. Data were merged across cohorts using QCTOOL and GTOOL, filtering out variants with MAF≤0.005). We performed two analyses: case‐only linear mixed‐model regression with AAO as outcome (LMM); and survival analysis using Cox proportional‐hazards frailty modeling (COXPHF) with time‐to‐AD‐onset (AAO in cases and censoring of controls at age‐at‐last‐exam). Both models were adjusted for sex, principal components capturing population substructure, and APOE‐ε4 dosage with a random cohort effect.ResultWe examined 2,262 AA AD cases, including 1,561 females (69%) with mean (SD) AAO of 77.7 (8.1) years. The APOE‐adjusted COXPHF analyses identified twelve genome‐wide significant regions (P<5×10−8), the most significant in ABCA7 (P = 3.57×10−10), known to be associated with AD risk in AAs. In the LMM analysis, we observed strong novel associations with variants in CCDC141 on chr2q31.2 (Pmin = 2.93×10−9). In silico analyses of LD structure and functional potential suggest SESTD1, a gene involved in synaptic regulation, as driving the CCDC141 locus signal. Nine of 12 COXPHF loci were driven by African‐specific variants (MAF<0.0002 in gnomAD Non‐Finish Europeans). These significant loci identified in the AA COXPHF or LMM analyses were not significant (P>0.05) in equivalent analyses in NHW ADGC cohorts (9,220 AD cases/10,350 controls; Li et al. Alz Dement 2023), except for the ABCA7 locus (NHW P<0.0002).ConclusionIn a large ADGC African American dataset, we confirmed AAO associations in the APOE region and ABCA7 and observed evidence for potential novel AD AAO loci including SESTD1; many of which might be African‐ancestry specific.

EVALUASI PEMBOROSAN MESIN JUMBO LOOM BERBASIS PENDEKATAN STATISTICAL PROCESS CONTROL

Continuous quality control along the production line is a crucial factor in achieving high productivity. PT. TRZ is a producer of various types of plastic-based products that implement strict quality control standards. However, it is still found that production targets still need to be achieved due to machine damage. This machine breaks. The company needs to be able to fulfill consumer orders on time. This research aims to evaluate product defects resulting from damage to the Jumbo Loom machine. Statistical process control evaluates factors in cutting machine defects, thread splice marks, and sheet shrinkage. The research results show that the company faces problems regarding machine defects, including machine running, thread splice marks, and shriveled sheets. This research aims to reduce defects in machines. The Seven Tools method is used to make repairs. From the processing results on the control chart, there were daily defects that exceeded the control limits. Therefore, it was carried out using a fishbone diagram. The processing results obtained 23 indicators of the causes of the three types of defects.

Polyphenol-enriched Penthorum chinense Pursh ameliorates alcohol-related liver injury through Ras/Raf/MEK/ERK pathway: Integrating network pharmacology and experiment validation

Ethnopharmacological relevancePenthorum chinense Pursh (PCP) has acknowledged as an edible herbal medicinal plant for the prevention and treatment of alcoholic liver injury (ALI). However, only few of researches focus on the chemical material basis and potential mechanisms of PCP against ALI. Aim of the studyHerein, we explored the therapeutic effects of PCP extract against ALI based on the integration of network pharmacology, molecular docking, and experiment validation. MethodsBased on the standard quality control of PCP herbs by UPLC fingerprint and quantitative determination, 80% ethanol extract fraction of PCP containing more polyphenols, compared to aqueous extract fraction of PCP, were chosen for further experiments. After oral administration of PCP ethanol extract, serum pharmacochemistry based on UPLC-Q-Exactive-MS analysis was implemented to evaluate the potential effective compounds. These absorbed prototypes in PCP were used to construct network pharmacology and predict the potential mechanisms of PCP extract against ALI. Then, the predicted targets and biological mechanisms of PCP extract were validated using animal experiments and molecular docking analysis. ResultsAlthough totally 19 polyphenol compounds were identified in PCP ethanol extract by UPLC-MS analysis, only 18 absorbed prototypes were found in the serum collected from mice at 1 h post-administration with PCP extract. These candidate active compounds were further screened into 13 compounds to construct network pharmacology and 433 targets were identified as PCP targets. GO and KEGG pathway enrichment analyses indicated that the effects of PCP extract would involve in Ras signaling pathway. The animal experiments on chronic ALI model mice shown that the oral administration of PCP can alleviate ALI by attenuating hepatic oxidative stress, inflammation and down-regulating the target proteins in Ras/Raf/MEK/ERK pathway. Molecular docking analysis revealed the good binding ability between the three polyphenols (i.e. quercetin, apigenin, thonningianin B) in PCP with the top contribution in network pharmacology, and these target proteins (Ras, Raf, MEK1/2, and ERK1/2). ConclusionOur results clarified that PCP ethanol extract could effectively alleviate ALI by down-regulating Ras/Raf/MEK/ERK signaling pathway promisingly. Quercetin, apigenin, and thonningianin B may be the active compounds of PCP, attributing to the intervention benefits of PCP against ALI.

Test-the-Test: Clinical Utility of Comprehensive Whole Exome Sequencing and RNA-Seq for Patients with Lymphoma

Introduction Due to the rapidly evolving landscape of targeted therapies, there is an unmet need for comprehensive molecular profiling to guide treatment decisions for patients with lymphoma. To meet this need, we designed a pilot study to assess the feasibility and turnaround time (TAT) of comprehensive whole exome (WES) and whole transcriptome (RNA-seq) sequencing (Tumor Portrait test) in patients with lymphoma for future clinical decision-making (NCT05464823). Here, we present interim results for patients with large B-cell lymphoma (LBCL). Methods Patients ≥18 years of age with histologically documented LBCL requiring therapy were eligible. Formalin-fixed paraffin-embedded tissues underwent WES, RNA-seq, and copy number analysis with concomitant peripheral blood or saliva for germline DNA sequencing. During quality control, samples with <20% tumor purity and median coverage of <142x for tumor and/or <95x for normal samples were labeled as quantity not sufficient (QNS)-qualified. Genomic and transcriptomic data were profiled for alterations and molecular features to uncover clinically relevant biomarkers and match patients with clinical trials using data from ClinicalTrials.gov. Results To date, 48 patients received WES and RNA-seq, and cohort data were analyzed for assay performance. Two samples were excluded from the analysis due to follicular lymphoma and chronic lymphocytic leukemia pathology, and seven samples were excluded as QNS. Reports were delivered for 39 patients, which included 37 clinical reports and two advanced reports lacking treatment and clinical trial information for QNS-qualified samples. While the QNS-qualified samples did not meet quality control standards, some data were still obtained and delivered as partial clinical reports. Of the 39 patients who received clinical reports, 34 samples were evaluable for bioinformatics analysis, and two of these samples had either only RNA-seq or WES data available (total n = 34, Figure 1). Cell of origin (COO), LymphGen (Wright et al. Cancer Cell. 2020), and lymphoma microenvironment (LME; Kotlov et al. Cancer Discov. 2021) classifications were applied to 33 patients. The median TAT was 8 days for advanced report delivery, and 85% of reports had a TAT <10 days. WES identified frequent TP53 (n = 11) and KMT2D (n = 10) loss of function and MYD88 (n = 5) gain of function alterations, as anticipated for LBCL. COOs were assigned as activated (ABC; n = 14, 42%) or germinal center (GCB; n = 19, 58%) LBCL. Using the LymphGen classifier, EZB MYC- (n = 7, 22%), A53 (n = 6, 19%), and MCD (n = 5, 16%) were the most prevalent subtypes. LME classification revealed the majority of samples were mesenchymal (ME; n = 22, 67%). Immune-depleted (DE; n = 6, 18%), immune-inflamed (IN; n = 4, 12%), and germinal center-like (GC; n = 1, 3%) subtypes were less prevalent, but the immune-depleted LME occurred more commonly in patients at relapse. Clinically significant findings, such as DHITsig+ (n = 4) and CARD11 mutations indicative of ibrutinib resistance (n = 3), were identified in 26 of the 39 patients ( Table 1), and an average of 8 clinical trials were identified for potential patient enrollment in each delivered report. Conclusions These data indicated that comprehensive WES and RNA-seq molecular profiling can be performed for lymphoma patients with acceptable TAT. This approach identified significant alterations and classified LBCL samples into previously reported lymphoma subtypes. The clinical reports included clinically relevant findings and matched clinical trials, demonstrating the feasibility of incorporating comprehensive molecular profiling in clinical practice for patients with lymphoma.

1020. A Novel Quality Control Solution for Gram Stain Through Integration of Deep Learning Vision Technology: A Proof-of-Concept

Abstract Background Gram stain (GS) is essential for preliminary bacterial identification. The lack of a standardized quality control method for GS is a significant drawback. By utilizing a deep convolutional neural network (CNN) design, we present a proof-of-concept for a novel solution to GS quality assurance. Using this approach, current manual GS, and potentially automated GS methods, may possess a standardized method of quality control. Methods Twenty-nine GS from positive blood cultures encompassing the spectrum of GS quality (acceptable, over/under decolorization, smear thickness, stain artifact) were selected by two expert microbiologists. A total of 197 images were obtained using a Cognex D-900 camera and Olympus BX-43 microscope at 1000X magnification. Images were classified using deep learning software (Cognex, ViDi Suite 4.1). Eighty-one images were classified as suboptimal and 116 were classified as optimal by expert review. To address background variation due to manual GS, we implemented a screening classification tool to separate images into 5 distinct classes (pale, rose, orange, indigo, variegated). The training and test sets consisted of 100 and 97 images respectively. Using a chaining technique, a second classification tool was added after the screening classification tool. The second tool further divided the 197 images into two dataset classes: optimal and suboptimal. The second tool used 105 images for training and 92 images for the test set. Model performance was compared to expert microbiology review, with GS verified by organism identification using MALDI-TOF. Results Cognex ViDi Suite 4.1 could discriminate between optimal and suboptimal GS images. The chained classification tool was able to be trained with 100% recall and 100% accuracy, with a resulting F-score of 1.00. Figure 1 VisionPro GUI with optimal image (MRSA) Conclusion Deep learning enabled image analysis is an effective method for sorting GS which are acceptable for further interpretation and suboptimal smears which require rework. Additional study is warranted to develop deep learning image analysis tools for the interpretation of GS on multiple specimen types and incorporate into automated GS systems. Disclosures Jane Hata, PhD , D(ABMM), Roche Molecular Systems: Grant/Research Support Kyle Magee, MLS(ASCP), SeluxDx: Employee

Evaluation of the Entrance Surface Doses (ESD) for common diagnostic X-ray examinations

ObjectiveThe purpose of this study was to assess the Entrance Skin Dose (ESD) in patients undergoing multiple types of radiographic examinations across three hospitals in Saudi Arabia. MethodsThe patient ESD was estimated directly using optically stimulated luminesces (OSL) and indirectly using a survey meter and calculations for 100 adult patients undergoing common diagnostic X-ray examinations, including a posterior anterior chest, posterior anterior hand, anterior posterior foot, and anterior posterior knee examinations, in three hospitals in Saudi Arabia. Comparative analysis with national and international reference values was conducted. ResultsOur analyses revealed considerable heterogeneity in X-ray exposure parameters among the hospitals. The mean ESD values for the chest examination were 0.28 ± 0.13 mGy for the direct method and 0.19 ± 0.11 mGy for the indirect method. Hand examination using the direct method resulted in a mean ESD of 0.42 ± 0.15 mGy, while the indirect method yielded 0.23 ± 0.06 mGy. Remarkably, the ESD values in most cases exceeded national and international guidelines, with the knee examination ESD using the direct and indirect methods showing an alarming value of 1.54 ± 0.31 mGy and 1.18 ± 0.21 mGy, respectively. The correlation between direct ESD, indirect ESD showed a strong positive correlation exists between direct ESD and Indirect ESD (r = 0.8, p < 0.001). ConclusionThe study highlights the urgent need for standardized protocols and quality control measures across radiographic facilities in Saudi Arabia. The elevated ESD values signify a critical issue that requires immediate attention to minimize patient radiation exposure while maintaining diagnostic image quality. Quality control tests and adjustments in examination parameters are strongly advised.

Multivariate Analysis among Marker Compounds, Environmental Factors, and Fruit Quality of Schisandra chinensis at Different Locations in South Korea.

This study aimed to investigate the correlation among the contents of marker compounds, growth characteristics, and environmental factors of Schisandra chinensis fruits across South Korea. The fruits were collected from 36 cultivation sites in 28 regions across the country. We investigated nine growth characteristics, twelve soil physicochemical properties, eight meteorological data, and three marker compounds in this study. We optimized and validated an optimized method for quantifying marker compounds using UPLC and performed correlation analysis among the contents of marker compounds, growth characteristics, and environmental factors. The UPLC-UV method for analyzing marker compounds was validated by measuring linearity, LOD, LOQ, precision, and accuracy. The marker compounds were negatively correlated with the fruit size and sugar contents, and growth characteristics were negatively correlated with some physicochemical properties of the soil. The results of this study can be used as basic data for the standard cultural practices and quality control of S. chinensis fruits.

Large-scale Normative Modeling of Brain Microstructure.

Normative models of brain metrics based on large populations are extremely valuable for detecting brain abnormalities in patients with dementia, psychiatric, or developmental conditions. Here we present the first large-scale normative model of the brain's white matter (WM) microstructure derived from 18 international diffusion MRI (dMRI) datasets covering almost the entire lifespan (totaling N=51,830 individuals; age: 3-80 years). We extracted regional diffusion tensor imaging (DTI) metrics using a standardized analysis and quality control protocol, and used Hierarchical Bayesian Regression (HBR) to model the statistical distribution of derived WM metrics as a function of age and sex, while modeling the site effect. HBR overcomes known weaknesses of some data harmonization methods that simply scale and shift residual distributions at each site. To illustrate the method, we applied it to detect and visualize profiles of WM microstructural deviations in cohorts of patients with Alzheimer's disease, mild cognitive impairment, Parkinson's disease and in carriers of 22q11.2 copy number variants, a rare neurogenetic condition that confers increased risk for psychosis. The resulting large-scale model offers a common reference to identify disease effects in individuals or groups, as well as to compare disorders and discover factors that influence these abnormalities.

Pharmaceutico-Analytical and Antimicrobial Study of Amritank Rasa

Amritank Rasa is a herbomineral formulation mentioned in the classical text of Basavrajiyam mainly used in the treatment of Pancha Kasa. This study aims to prepare the Amritank Rasa as per the classical text and conduct analytical and antimicrobial study of the prepared sample. The ingredients of Amritank Rasa are Kajjali, Trikatu, Pippalimoola, Chavya, Chitraka, Vatsanabha and Saindhava lavana all in equal parts and bhavana of Bhringaraj swarasa. The pharmaceutical procedure involves the preparation of Amritank Rasa vati in the 125mg dosage form. The pharmaceutical, analytical and antimicrobial parameters were compiled and data was recorded. The organoleptic parameters were, dark greyish in colour with pungent odour and taste, appearance was round and uncoated, and smooth in touch. The physiochemical parameters and quantitative parameters were, total ash was 5.5%, acid insoluble ash was 0.5% and loss on drying was 11.8%, average weight was 121mg, uniformity of weight test showed that all the tablets were not within the range of deviation from average weight, friability test was 0.51%, hardness was 5.3kg/cm2 and disintegration time test showed that any tablet did not dissolve in 60 minutes. The sample was then subjected to advanced analytical parameters i.e., FTIR, PSD, XRF and XRD. The antimicrobial study of Amritank rasa showed that it is effective against S.aureus, E.coli, P.aeruginosa and C. albicans. The development of the present study will serve as reference standards for Amritank Rasa formulation, quality control and clinical research

Improved Biorepository to Support Sickle Cell Disease Genomics and Clinical Research: A Practical Approach to Link Patient Data and Biospecimens from Muhimbili Sickle Cell Program, Tanzania.

In Africa, sickle cell disease phenotypes' genetic contributors remain understudied due to the dearth of databases that pair biospecimens with demographic and clinical details. The absence of biorepositories in these settings can exacerbate this issue. This article documents the physical verification process of biospecimens in the biorepository, connecting them to patient clinical and demographic data and aiding in the planning of future genomic and clinical research studies' experience from the Muhimbili Sickle Cell Program in Dar es Salaam, Tanzania. The biospecimen database was updated with the current biospecimen position following the physical verification and then mapping this information to its demographic and clinical data using demographic identifiers. The biorepository stored 74,079 biospecimens in three -80°C freezers, including 63,345 from 5159 patients enrolled in the cohort between 2004 and 2016. Patients were identified by a control (first visit), entry (when confirmed sickle cell homozygous), admission (when hospitalized), and follow-up numbers (subsequent visits). Of 63,345 biospecimens, follow-ups were 46,915 (74.06%), control 8067 (12.74%), admission 5517 (8.71%), and entry 2846 (4.49%). Of these registered patients, females were 2521 (48.87%) and males were 2638 (51.13%). The age distribution was 1-59 years, with those older than 18 years being 577 (11.18%) and children 4582 (88.82%) of registered patients. The notable findings during the process include a lack of automated biospecimen checks, laboratory information management system, and tubes with volume calibration; this caused the verification process to be tedious and manual. Biospecimens not linked to clinical and demographic data, date format inconsistencies, and lack of regular updating of a database on exhausted biospecimens and updates when biospecimens are moved between positions within freezers were other findings that were found. A well-organized biorepository plays a crucial role in answering future research questions. Enforcing standard operating procedures and quality control will ensure that laboratory users adhere to the best biospecimen management procedures.