Automated and robust extraction of genomic DNA from various leftover blood samples.

With the development of genomic technologies, the isolation of genomic DNA (gDNA) from clinical samples is increasingly required for clinical diagnostics and research studies. In this study, we explored the potential of utilizing various leftover blood samples obtained from routine clinical tests as a viable source of gDNA. Using an automated method with optimized pre-treatments, we obtained gDNA from seven types of clinical leftover blood, with average yields of gDNA ranging from 3.11 ± 0.45 to 22.45 ± 4.83 µg. Additionally, we investigated the impact of storage conditions on gDNA recovery, resulting in yields of 8.62-68.08 µg when extracting gDNA from EDTA leftover blood samples stored at 4 °C for up to 13 weeks or -80 °C for up to 78 weeks. Furthermore, we successfully obtained sequenceable gDNA from both Serum Separator Tube and EDTA Tube using a 96-well format extraction, with yields ranging from 0.61 to 71.29 µg and 3.94-215.98 µg, respectively. Our findings demonstrate the feasibility of using automated high-throughput platforms for gDNA extraction from various clinical leftover blood samples with the proper pre-treatments.

Antioxidants in Traditional Mexican Medicine and Their Applications as Antitumor Treatments.

Traditional medicine in Latin America and mainly in Mexico represents an essential alternative for treating different diseases. The use of plants as medicine is the product of a rich cultural tradition of the indigenous peoples, in which a great variety of species are used for the treatment of gastrointestinal, respiratory, and mental diseases and some other sicknesses; the therapeutic efficacy that they possess is due to the properties that derive from the active ingredients of plants principally antioxidants, such as phenolic compounds, flavonoids, terpenes, and tannins. An antioxidant is a substance that, at low concentrations, delays or prevents substrate oxidation through the exchange of electrons. Different methods are used to determine the antioxidant activity and the most commonly used are described in the review. Cancer is a disease in which some cells multiply uncontrollably and spread to other parts of the body, a process known as metastasis. These cells can lead to the formation of tumors, which are lumps of tissue that can be cancerous (malignant) or noncancerous (benign). Generally, the treatment of this disease consists of surgery, radiotherapy, or chemotherapy, which have side effects that decrease the quality of life of patients, so new treatments, focusing on natural resources such as plants, can be developed. This review aims to gather scientific evidence on the antioxidant compounds present in plants used in traditional Mexican medicine, specifically as antitumor treatment in the most common cancer types worldwide (e.g., breast, liver, and colorectal cancer).

The Paradigm of the Inflammatory Radicular Cyst: Biological Aspects to be Considered.

Inflammatory radicular cysts (IRCs) are chronic lesions that follow the development of periapical granulomas (PGs). IRCs result from multiple inflammatory reactions led initially by several pro-inflammatory interleukins and growth factors that provoke the proliferation of epithelial cells derived from epithelial cell rests of Malassez present in the granulomatous tissue, followed by cyst formation and growth processes. Multiple theories have been proposed to help explain the molecular process involved in the development of the IRC from a PG. However, although multiple studies have demonstrated the presence of epithelial cells in most PGs, it is still not fully understood why not all PGs turn into IRCs, even though both are stages of the same inflammatory phenomenon and receive the same antigenic stimulus. Histopathological examination is currently the diagnostic gold standard for differentiating IRCs from PGs. Although multiple studies have evaluated the accuracy of non-invasive or minimally invasive methods in assessing the histopathological nature of the AP before the intervention, these studies' results are still controversial. This narrative review addresses the biological insights into the complex molecular mechanisms of IRC formation and its histopathological features. In addition, the relevant inflammatory molecular mediators for IRC development and the accuracy of non-invasive or minimally invasive diagnostic approaches are summarised. (EEJ-2022-03-041).

Protocol for Biospecimen Collection and Analysis Within the BACPAC Research Program.

The Biospecimen Collection and Processing Working Group of the National Institutes of Health (NIH) HEAL Initiative BACPAC Research Program was charged with identifying molecular biomarkers of interest to chronic low back pain (cLBP). Having identified biomarkers of interest, the Working Group worked with the New York University Grossman School of Medicine, Center for Biospecimen Research and Development-funded by the Early Phase Pain Investigation Clinical Network Data Coordinating Center-to harmonize consortium-wide and site-specific efforts for biospecimen collection and analysis. Biospecimen collected are saliva, blood (whole, plasma, serum), urine, stool, and spine tissue (paraspinal muscle, ligamentum flavum, vertebral bone, facet cartilage, disc endplate, annulus fibrosus, or nucleus pulposus). The omics data acquisition and analyses derived from the biospecimen include genomics and epigenetics from DNA, proteomics from protein, transcriptomics from RNA, and microbiomics from 16S rRNA. These analyses contribute to the overarching goal of BACPAC to phenotype cLBP and will guide future efforts for precision medicine treatment.

Broad antiviral spectrum of glycyrrhizic acid for human and veterinary medicine: reality or fiction?

BACKGROUND: Emerging virus infections provoke health problems in people and animals, which generate social, and economic issues worldwide. This has spurred the search for new pharmacological strategies to confront them. SUMMARY: The purpose of this review is to draw the reader's attention to pharmacological evaluations of glycyrrhizic acid (GA) and its analogs on the broad range of viruses known in human and veterinary medicine. GA is the main water-soluble constituent extracted from the roots of plants from the genus Glycyrrhiza, commonly known as licorice root. It has long been used due to its broad spectrum of bioactivities, including anti-inflammatory, antiulcer, and antitumor properties. It has also been proposed as an antiviral agent. Medicines derived from GA are currently being used to combat acute and chronic hepatitis and herpes viruses. KEY MESSAGES: This review suggests that GA could be a new broad-spectrum antiviral due to its ability to inhibit DNA or RNA viruses both in vitro and in vivo. GA could be a potential drug for preventing and/or treating various viral diseases.

Potential Anticancer Activity of Pomegranate (Punica granatum L.) Fruits of Different Color: In Vitro and In Silico Evidence.

Pomegranate (PMG; Punica granatum L.) fruits possess a well-balanced nutrient/phytochemical composition, with proven adjuvant benefits in experimental cancer chemotherapy; however, such bioactivity could be affected by PMG's phenogenotype (varietal). Here, the chemical and phytochemical (UPLC-DAD-MS2) composition, antioxidant capacity and anticancer potential [in vitro (MTT assay) and in silico (foodinformatics)] of three PMG fruits of different aryl color [red (cv. Wonderful), pink (cv. Molar de Elche), and white (cv. Indian)] were evaluated. The macro/micronutrient (ascorbic acid, tocols, carotenoids), organic acid (citric/malic), and polyphenol content were changed by PMG's varietal and total antioxidant activity (ABTS, alcoholic > hexane extract) in the order of red > pink > white. However, their in vitro cytotoxicity was the same (IC50 > 200 µg.mL-1) against normal (retinal) and cancer (breast, lung, colorectal) cell lines. Sixteen major phytochemicals were tentatively identified, four of them with a high GI absorption/bioavailability score [Ellagic (pink), vanillic (red), gallic (white) acids, D-(+)-catechin (white)] and three of them with multiple molecular targets [Ellagic (52) > vanillic (32) > gallic (23)] associated with anticancer (at initiation and promotion stages) activity. The anticancer potential of the PMG fruit is phenogenotype-specific, although it could be more effective in nutraceutical formulations (concentrates).

Gold Nanoparticles Synthesized with Common Mullein (Verbascum thapsus) and Castor Bean (Ricinus communis) Ethanolic Extracts Displayed Antiproliferative Effects and Induced Caspase 3 Activity in Human HT29 and SW480 Cancer Cells.

Gold nanoparticles (AuNPs) are promising nanomaterials exhibiting anti-cancer effects. Green AuNPs synthesis using plant extracts can be used to achieve stable and beneficial nanoparticles due to their content of bioactive compounds. This research aimed to synthesize and evaluate the antiproliferative and caspase-3 activity induction of green AuNPs synthesized with common mullein (V. thapsus) flowers (AuNPsME) and castor bean (R. communis) leaves (AuNPsCE) ethanolic extracts in human HT29 and SW480 colorectal cancer cells. Their effect was compared with chemically synthesized AuNPs (AuNPsCS). The extracts mainly contained p-coumaric acid (71.88-79.93 µg/g), ferulic acid (19.07-310.71 µg/g), and rutin (8.14-13.31 µg/g). The obtained nanoparticles presented typical FT-IR bands confirming the inclusion of polyphenols from V. thapsus and R. communis and spherical/quasi-spherical morphologies with diameters in the 20.06-37.14 nm range. The nanoparticles (20-200 µg/mL) showed antiproliferative effects in both cell lines, with AuNPsCE being the most potent (IC50 HT29: 110.10 and IC50SW480: 64.57 µg/mL). The AuNPsCS showed the lowest intracellular reactive oxygen species (ROS) generation in SW480 cells. All treatments induced caspase 3/7 activity to a similar or greater extent than 30 mM H2O2-treated cells. Results indicated the suitability of V. thapsus and R. communis extracts to synthesize AuNPs, displaying a stronger antiproliferative effect than AuNPsCS.

Safety and immunogenicity of an AS03-adjuvanted SARS-CoV-2 recombinant protein vaccine (CoV2 preS dTM) in healthy adults: interim findings from a phase 2, randomised, dose-finding, multicentre study.

BACKGROUND: We evaluated our SARS-CoV-2 prefusion spike recombinant protein vaccine (CoV2 preS dTM) with different adjuvants, unadjuvanted, and in a one-injection and two-injection dosing schedule in a previous phase 1-2 study. Based on interim results from that study, we selected a two-injection schedule and the AS03 adjuvant for further clinical development. However, lower than expected antibody responses, particularly in older adults, and higher than expected reactogenicity after the second vaccination were observed. In the current study, we evaluated the safety and immunogenicity of an optimised formulation of CoV2 preS dTM adjuvanted with AS03 to inform progression to phase 3 clinical trial. METHODS: This phase 2, randomised, parallel-group, dose-ranging study was done in adults (≥18 years old), including those with pre-existing medical conditions, those who were immunocompromised (except those with recent organ transplant or chemotherapy) and those with a potentially increased risk for severe COVID-19, at 20 clinical research centres in the USA and Honduras. Women who were pregnant or lactating or, for those of childbearing potential, not using an effective method of contraception or abstinence, and those who had received a COVID-19 vaccine, were excluded. Participants were randomly assigned (1:1:1) using an interactive response technology system, with stratification by age (18-59 years and ≥60 years), rapid serodiagnostic test result (positive or negative), and high-risk medical conditions (yes or no), to receive two injections (day 1 and day 22) of 5 7mu;g (low dose), 10 7mu;g (medium dose), or 15 7mu;g (high dose) CoV2 preS dTM antigen with fixed AS03 content. All participants and outcome assessors were masked to group assignment; unmasked study staff involved in vaccine preparation were not involved in safety outcome assessments. All laboratory staff performing the assays were masked to treatment. The primary safety objective was to describe the safety profile in all participants, for each candidate vaccine formulation. Safety endpoints were evaluated for all randomised participants who received at least one dose of the study vaccine (safety analysis set), and are presented here for the interim study period (up to day 43). The primary immunogenicity objective was to describe the neutralising antibody titres to the D614G variant 14 days after the second vaccination (day 36) in participants who were SARS-CoV-2 naive who received both injections, provided samples at day 1 and day 36, did not have protocol deviations, and did not receive an authorised COVID-19 vaccine before day 36. Neutralising antibodies were measured using a pseudovirus neutralisation assay and are presented here up to 14 days after the second dose. As a secondary immunogenicity objective, we assessed neutralising antibodies in non-naive participants. This trial is registered with ClinicalTrials.gov (NCT04762680) and is closed to new participants for the cohort reported here. FINDINGS: Of 722 participants enrolled and randomly assigned between Feb 24, 2021, and March 8, 2021, 721 received at least one injection (low dose=240, medium dose=239, and high dose=242). The proportion of participants reporting at least one solicited adverse reaction (injection site or systemic) in the first 7 days after any vaccination was similar between treatment groups (217 [91%] of 238 in the low-dose group, 213 [90%] of 237 in the medium-dose group, and 218 [91%] of 239 in the high-dose group); these adverse reactions were transient, were mostly mild to moderate in intensity, and occurred at a higher frequency and intensity after the second vaccination. Four participants reported immediate unsolicited adverse events; two (one each in the low-dose group and medium-dose group) were considered by the investigators to be vaccine related and two (one each in the low-dose and high-dose groups) were considered unrelated. Five participants reported seven vaccine-related medically attended adverse events (two in the low-dose group, one in the medium-dose group, and four in the high-dose group). No vaccine-related serious adverse events and no adverse events of special interest were reported. Among participants naive to SARS-CoV-2 at day 36, 158 (98%) of 162 in the low-dose group, 166 (99%) of 168 in the medium-dose group, and 163 (98%) of 166 in the high-dose group had at least a two-fold increase in neutralising antibody titres to the D614G variant from baseline. Neutralising antibody geometric mean titres (GMTs) at day 36 for participants who were naive were 2189 (95% CI 1744-2746) for the low-dose group, 2269 (1792-2873) for the medium-dose group, and 2895 (2294-3654) for the high-dose group. GMT ratios (day 36: day 1) were 107 (95% CI 85-135) in the low-dose group, 110 (87-140) in the medium-dose group, and 141 (111-179) in the high-dose group. Neutralising antibody titres in non-naive adults 21 days after one injection tended to be higher than titres after two injections in adults who were naive, with GMTs 21 days after one injection for participants who were non-naive being 3143 (95% CI 836-11 815) in the low-dose group, 2338 (593-9226) in the medium-dose group, and 7069 (1361-36 725) in the high-dose group. INTERPRETATION: Two injections of CoV2 preS dTM-AS03 showed acceptable safety and reactogenicity, and robust immunogenicity in adults who were SARS-CoV-2 naive and non-naive. These results supported progression to phase 3 evaluation of the 10 7mu;g antigen dose for primary vaccination and a 5 7mu;g antigen dose for booster vaccination. FUNDING: Sanofi Pasteur and Biomedical Advanced Research and Development Authority.

Effect on plant growth parameters and secondary metabolite content of lettuce (Lactuca sativa L.), coriander (Coriandrum sativum), and chili pepper (Capsicum annuum L.) watered with disinfected water by Ag-TiO2 nanoparticles.

Nowadays, the use of different nanoscale structures has been introduced to a large number of research areas. One of these is the treatment and remediation of water through photocatalytic processes, seeking to reuse wastewater for agriculture. In this paper, Lactuca sativa, Coriandrum sativum, and Capsicum annuum were used as crop models to observe the effects in plant growth and the secondary metabolism of different water qualities and types used in the watering process. Initial results show that the photocatalytic process's water maintains a pH and ion concentration within the allowed limits, significantly reducing the number of bacteria. Along the growth process, an influence on germination times, appearance of true leaves, maturation, and fruit production depending on the type of water used is observed, obtaining the best results in both growth times and quantity of fruits, for the 50% and 70% disinfected water/tap water (DW/TAW) study groups. Secondary metabolites, such as phenols, flavonoids, and antioxidant activity, were studied to evaluate changes in the vegetables' composition, showing increased concentration for the disinfected water groups in most specimens. Additionally, no traces of metals and microorganisms were detected, concluding that the crops are viable to be consumed by human beings.

Strategies to Improve the Properties of Amaranth Protein Isolate-Based Thin Films for Food Packaging Applications: Nano-Layering through Spin-Coating and Incorporation of Cellulose Nanocrystals.

In this work, two different strategies for the development of amaranth protein isolate (API)-based films were evaluated. In the first strategy, ultrathin films were produced through spin-coating nanolayering, and the effects of protein concentration in the spin coating solution, rotational speed, and number of layers deposited on the properties of the films were evaluated. In the second strategy, cellulose nanocrystals (CNCs) were incorporated through a casting methodology. The morphology, optical properties, and moisture affinity of the films (water contact angle, solubility, water content) were characterized. Both strategies resulted in homogeneous films with good optical properties, decreased hydrophilic character (as deduced from the contact angle measurements and solubility), and improved mechanical properties when compared with the neat API-films. However, both the processing method and film thickness influenced the final properties of the films, being the ones processed through spin coating more transparent, less hydrophilic, and less water-soluble. Incorporation of CNCs above 10% increased hydrophobicity, decreasing the water solubility of the API films and significantly enhancing material toughness.

Evaluation and modeling of changes in color, firmness, and physicochemical shelf life of cut pineapple (Ananas comosus) slices in equilibrium-modified atmosphere packaging.

In this study cut, pineapple slices of 1 cm thick were packaged and stored at different temperatures and equilibrium modified atmosphere packages (EMAPs) to determine changes of color and firmness over time to represent physicochemical shelf life. From the experimental data, a variance analysis was performed to determine the effect of temperature and O2 level on the evolution of color (CIELAB coordinates) and firmness. It was observed that the evolution in L* , a* , and b* coordinates is independent on O2 concentration in the EMAP system. After that, suitable models were adjusted to represent the change of the quality properties as a function of temperature by using first-order models to represent color and a power model for firmness. Likewise, a modified normal distribution function was adjusted to represent the coefficient of firmness loss depending on the O2 level besides the temperature effect. The firmness model was used to obtain a suitable equation to predict shelf life of the pineapple slices for different EMAP systems. Finally, a validation experiment was performed at 8 °C obtaining a high capacity of prediction (R2 adj > 0.90) compared with the experimental data. The adjusted model can be used to configure a satisfactory EMAP system for the best preservation of minimally processed pineapple from the predicted evolution of color, firmness, and shelf life depending on temperature and O2 concentration. PRACTICAL APPLICATION: In this work, we built a mathematical model to simulate the shelf life of pineapple cut into slices based on changes in color and firmness and as a function of temperature and oxygen level. These properties are truly relevant because they are very clear evidence of the fruit deterioration, and for that reason, we chose them for the model. The model we developed can be applied in retail and supermarket systems to determine precisely how long the product on the shelf can last before being disposed of, reducing material losses.

A Low-Cost Visible Light Positioning System for Indoor Positioning.

Currently, a high percentage of the world's population lives in urban areas, and this proportion will increase in the coming decades. In this context, indoor positioning systems (IPSs) have been a topic of great interest for researchers. On the other hand, Visible Light Communication (VLC) systems have advantages over RF technologies; for instance, they do not need satellite signals or the absence of electromagnetic interference to achieve positioning. Nowadays, in the context of Indoor Positioning (IPS), Visible Light Positioning (VLP) systems have become a strong alternative to RF-based systems, allowing the reduction in costs and time to market. This paper shows a low cost VLP solution for indoor systems. This includes multiple programmable beacons and a receiver which can be plugged to a smartphone running a specific app. The position information will be quickly and securely available through the interchange between the receiver and any configurable LED-beacon which is strategically disposed in an area. The implementation is simple, inexpensive, and no direct communication with any data server is required.

Nanoparticles in Agroindustry: Applications, Toxicity, Challenges, and Trends.

Nanotechnology is a tool that in the last decade has demonstrated multiple applications in several sectors, including agroindustry. There has been an advance in the development of nanoparticulated systems to be used as fertilizers, pesticides, herbicides, sensors, and quality stimulants, among other applications. The nanoencapsulation process not only protects the active ingredient but also can affect the diffusion, interaction, and activity. It is important to evaluate the negative aspects of the use of nanoparticles (NPs) in agriculture. Given the high impact of the nanoparticulated systems in the agro-industrial field, this review aims to address the effects of various nanomaterials on the morphology, metabolomics, and genetic modification of several crops.

The urgency of utilizing COVID-19 biospecimens for research in the heart of the global pandemic.

The outbreak of the novel coronavirus disease 2019 (COVID-19) and consequent social distancing practices have disrupted essential clinical research functions worldwide. Ironically, this coincides with an immediate need for research to comprehend the biology of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the pathology of COVID-19. As the global crisis has already led to over 15,000 deaths out of 175,000 confirmed cases in New York City and Nassau County, NY alone, it is increasingly urgent to collect patient biospecimens linked to active clinical follow up. However, building a COVID-19 biorepository amidst the active pandemic is a complex and delicate task. To help facilitate rapid, robust, and regulated research on this novel virus, we report on the successful model implemented by New York University Langone Health (NYULH) within days of outbreak in the most challenging hot spot of infection globally. Using an amended institutional biobanking protocol, these efforts led to accrual of 11,120 patients presenting for SARS-CoV-2 testing, 4267 (38.4%) of whom tested positive for COVID-19. The recently reported genomic characterization of SARS-CoV-2 in the New York City Region, which is a crucial development in tracing sources of infection and asymptomatic spread of the novel virus, is the first outcome of this effort. While this growing resource actively supports studies of the New York outbreak in real time, a worldwide effort is necessary to build a collective arsenal of research tools to deal with the global crisis now, and to exploit the virus's biology for translational innovation that outlasts humanity's current dilemma.

First-Pass Metabolism of Polyphenols from Selected Berries: A High-Throughput Bioanalytical Approach.

Small berries are rich in polyphenols whose first-pass metabolism may alter their ultimate physiological effects. The antioxidant capacity and polyphenol profile of three freeze-dried berries (blackberry, raspberry, Red Globe grape) were measured and their apparent permeability (Papp) and first-pass biotransformation were tracked with an ex vivo bioanalytical system [everted gut sac (rat) + three detection methods: spectrophotometry, HPLC-ESI-QTOF-MS, differential pulse voltammetry (DPV)]. Total polyphenol (ratio 0.07-0.14-1.0) and molecular diversity (anthocyanins>flavan-3-ols), antioxidant capacity (DPPH, FRAP), anodic current maxima and Papp (efflux> uptake) were in the following order: blackberry > raspberry > Red Globe grape. Epicatechin, pelargonidin & cyanin (all), callistephin (raspberry/blackberry), catechin (grape), cyanidin glycosides (blackberry) and their derived metabolites [quinic acid, epicatechin, cyanidin/malvidin glucosides, and chlorogenic/caffeic acids] were fruit-specific and concentration-dependent. Time-trend DPV kinetic data revealed concurrent epithelial permeability & biotransformation processes. Regular permeability and high-biotransformation of berry polyphenols suggest fruit-specific health effects apparently at the intestinal level.

Chemical characterization, antioxidant and antimutagenic evaluations of pigmented corn.

In order to identify pigmented corn with nutraceutical potential, the secondary metabolite content, the antioxidant capacity and antimutagenic activity of red, and blue corn were analyzed. The ranges of total phenolic, flavonoid and anthocyanin contents of the corn samples were from 69.4 to 212.8 mg gallic ac. equiv./100 g DW, 0.07 to 12.19 mg (+) catechin eq./100 g DW and 3.89 to 34.17 mg cyanidin-3-O-glucoside eq./100 g DW, respectively. The phenolic extracts demonstrated the highest antioxidant capacity evaluated by the ABTS assay displaying values from 2.06 to 7.34 mmol Trolox/100 g DW. None of the extracts was toxic to the tested bacteria strains TA98 and TA100. For TA98 tester strain, percentage inhibition values against AFB1 mutagenicity from 61 to 93, and 38 to 75 for flavonoid and anthocyanin extracts were obtained. The total phenol and anthocyanin contents correlate with the observed antioxidant capacity. The most biological active corn samples were the blue color while the least actives were the red ones. The results show that the studied blue corn samples are good sources of antioxidant and antimutagenic compounds, which could use to develop products that contribute to human health.

Antimicrobial effect of nisin electrospun amaranth: pullulan nanofibers in apple juice and fresh cheese.

Nisin-loaded amaranth protein isolate:pullulan (API:PUL) nanofibers were prepared by the electrospinning method. The nisin release kinetic was evaluated at pH 3.4 and 6.1 and the antimicrobial effectiveness of the electrospun mats was evaluated in apple juice and fresh cheese. The nisin API:PUL fibers with 120 nm average diameter reached 81.49% and 43.85% nisin release after 12 h at pH 3.4 and 6.1, respectively. The encapsulation of nisin in electrospun fibers allowed complete bactericidal activity against Salmonella Typhimurium, L. monocytogenes and L. mesenteroides inoculated in apple juice after 48, 20 and 48 h, respectively. When nisin API:PUL fibers were applied to fresh cheese, microorganism inactivation was complete after 142, 120 and 170 h, respectively. The results demonstrated that nisin API:PUL electrospun fibers significantly reduce the bacterial population and can be used in food products for microbiological safety.

Interlaboratory proficiency processing scheme in CSF aliquoting: implementation and assessment based on biomarkers of Alzheimer's disease.

BACKGROUND: In this study, we tested to which extent possible between-center differences in standardized operating procedures (SOPs) for biobanking of cerebrospinal fluid (CSF) samples influence the homogeneity of the resulting aliquots and, consequently, the concentrations of the centrally analyzed selected Alzheimer's disease biomarkers. METHODS: Proficiency processing samples (PPSs), prepared by pooling of four individual CSF samples, were sent to 10 participating centers, which were asked to perform aliquoting of the PPSs into two secondary aliquots (SAs) under their local SOPs. The resulting SAs were shipped to the central laboratory, where the concentrations of amyloid beta (Aß) 1-42, pTau181, and albumin were measured in one run with validated routine analytical methods. Total variability of the concentrations, and its within-center and between-center components, were analyzed with hierarchical regression models. RESULTS: We observed neglectable variability in the concentrations of pTau181 and albumin across the centers and the aliquots. In contrast, the variability of the Aß1-42 concentrations was much larger (overall coefficient of variation 31%), with 28% of the between-laboratory component and 10% of the within-laboratory (i.e., between-aliquot) component. We identified duration of the preparation of the aliquots and the centrifugation force as two potential confounders influencing within-center variability and biomarker concentrations, respectively. CONCLUSIONS: Proficiency processing schemes provide objective evidence for the most critical preanalytical variables. Standardization of these variables may significantly enhance the quality of the collected biospecimens. Studies utilizing retrospective samples collected under different local SOPs need to consider such differences in the statistical evaluations of the data.