Identification Protocol Research Articles

ASSESSMENT OF Staphylococcus aureus PREVALENCE IN VEGETABLES SOLD AT LOCAL MARKETS IN BENIN CITY: IMPLICATIONS FOR FOOD SAFETY.

Vegetables are an essential part of people's diet globally but they could aid the transmission of potential pathogens and channel the dissemination of resistance genes. This study aimed at evaluating the prevalence and antibiogram profile of Staphylococcus aureus in vegetables obtained from local markets in Benin City, Nigeria. A total of 50 fresh vegetables were collected from five local markets and investigated. These include 15 green leafs (Amaranthus tricolor), 15 water leafs (Talinum fruticosum), and 20 pumpkin leafs (Telfairia occidentalis). The samples were screened for Staphylococcus aureus using cultural, biochemical and Analytical Profile Index (API) Staph 32 identification protocols. The phenotypic characterization of antibiotic resistance was evaluated using the Kirby-Bauer disc diffusion technique. The distribution of Staphylococcus aureus as observed was green leafs [3/14 (21.4%)], water leafs [6/14 (42.9%)] and pumpkin leafs [5/14 (35.7%)]. The distribution of Staphylococcus aureus based on market location was showed the highest occurrence at the Oba market (40%) and New Benin market (40%) while the least occurrence was observed at New-market (10%). Overall, the total Staphylococcus aureus positive samples were 14/50 (28.0%) in all markets. The antibiotic-resistance profile of Staphylococcus aureus demonstrated by the isolates showed that the highest resistance was recorded for penicillin while no resistance was observed for nitrofurantoin and gentamicin. The multiple antibiotic resistance (MAR) index of Staphylococcus aureus in this study ranged from 0.14 – 0.71. Findings from this study and observed retail practices in local markets ascertained the possibility of vegetables getting contaminated via unhygienic agronomic and food handling practices.

Abstract 16045: Fluoroless Arterial Access and Closure for VT Ablation: Feasibility and Safety of an Ultrasound Guided Landmark Approach

Introduction: Ultrasound (US) guided access has become the gold standard of care for central access cannulation. The FAUST1 trial demonstrated that routine real-time US guidance was equivalent to fluoroscopic guidance but improved common femoral artery (CFA) access success and reduced complications in patients with high CFA bifurcations. The utilization of non radiation imaging modalities over the past decade has resulted in significantly reduced radiation exposure and orthopedic problems associated with prolonged electrophysiology procedures2. Hypothesis: Why do many operators still use fluoroscopy and arteriograms in addition to ultrasound guided access? Methods: A prospective single center multi-operator study was performed on patients undergoing premature ventricular contractions (PVC) or Ventricular Tachycardia (VT) ablation with arterial access. CFA access was obtained using a US guided landmark identification protocol. US probe is utilized in short and long access to identify upper and lower borders of the femoral head in relation to the CFA bifurcation. CFA access is then obtained with a modified Seldinger approach within femoral head landmarks. After arterial sheath insertion, the artery is again evaluated for closure appropriateness, assessed by absence of arterial calcifications within 5mm of arrteriotomy or stenosis <5mm in diameter. Results: 20 consecutive patients undergoing PVC or PVT ablation were included. All patients underwent US only guided access to the right common femoral artery. All patients underwent successful closure with a hemostasis device. There were no vascular complications. Conclusions: Real-time US only guided CFA access is feasible and safe. Appropriate utilization of US is sufficient to avoid intravenous contrast and radiation exposure without increased complication rate.

Unified Identification Protocol for Cross-Border Healthcare.

The Unified Identification Protocol (UIP) is an innovation which empowers patients and legal guardians to generate their unique digital identity for cross-border healthcare. This digital identity seamlessly links to local identifiers across different territories and organizations, bridging the gap between disparate systems. Combined with the International Patient Summary (IPS) - endorsed by the G7 and the EU - UIP is pioneering a new paradigm in telehealth services. Championing a user-centric approach in line with Web 3.0 principles, UIP places data control directly in the hands of patients and their legal guardians. This ensures accurate identification, streamlined access to health data, and robust privacy protection. When harmonized with tools like the SMART-On-FHIR API, FHIR Contract, DID Documents, and blockchain certification, UIP lays down transparent, user-approved guidelines for sharing healthcare data across borders. This framework guarantees that data is securely exchanged, encrypted specifically for the intended recipients upon user consent, adhering to international standards, and in full compliance with prevailing regulations. Furthermore, UIP facilitates certification of health courses and competences for patients, caregivers, and practitioners, enhancing healthcare understanding and management.

Future Perspectives for the Identification and Sequencing of Nicotinamide Adenine Dinucleotide-Capped RNAs.

Ribonucleic acid (RNA) is composed primarily of four canonical building blocks. In addition, more than 170 modifications contribute to its stability and function. Metabolites like nicotinamide adenine dinucleotide (NAD) were found to function as 5'-cap structures of RNA, just like 7-methylguanosine (m7G). The identification of NAD-capped RNA sequences was first made possible by NAD captureSeq, a multistep protocol for the specific targeting, purification, and sequencing of NAD-capped RNAs, developed in the authors' laboratory in the year 2015. In recent years, a number of NAD-RNA identification protocols have been developed by researchers around the world. They have enabled the discovery and identification of NAD-RNAs in bacteria, archaea, yeast, plants, mice, and human cells, and they play a key role in studying the biological functions of NAD capping. We introduce the four parameters of yield, specificity, evaluability, and throughput and describe to the reader how an ideal NAD-RNA identification protocol would perform in each of these disciplines. These parameters are further used to describe and analyze existing protocols that follow two general methodologies: the capture approach and the decapping approach. Capture protocols introduce an exogenous moiety into the NAD-cap structure in order to either specifically purify or sequence NAD-capped RNAs. In decapping protocols, the NAD cap is digested to 5'-monophosphate RNA, which is then specifically targeted and sequenced. Both approaches, as well as the different protocols within them, have advantages and challenges that we evaluate based on the aforementioned parameters. In addition, we suggest improvements in order to meet the future needs of research on NAD-modified RNAs, which is beginning to emerge in the area of cell-type specific samples. A limiting factor of the capture approach is the need for large amounts of input RNA. Here we see a high potential for innovation within the key targeting step: The enzymatic modification reaction of the NAD-cap structure catalyzed by ADP-ribosyl cyclase (ADPRC) is a major contributor to the parameters of yield and specificity but has mostly seen minor changes since the pioneering protocol of NAD captureSeq and needs to be more stringently analyzed. The major challenge of the decapping approach remains the specificity of the decapping enzymes, many of which act on a variety of 5'-cap structures. Exploration of new decapping enzymes or engineering of already known enzymes could lead to improvements in NAD-specific protocols. The use of a curated set of decapping enzymes in a combinatorial approach could allow for the simultaneous detection of multiple 5'-caps. The throughput of both approaches could be greatly improved by early sample pooling. We propose that this could be achieved by introducing a barcode RNA sequence before or immediately after the NAD-RNA targeting steps. With increased processing capacity and a potential decrease in the cost per sample, protocols will gain the potential to analyze large numbers of samples from different growth conditions and treatments. This will support the search for biological roles of NAD-capped RNAs in all types of organisms.

30,000 years of fishing in the Philippines: New ichthyoarchaeological investigations in Occidental Mindoro

Marine adaptations are considered to have been significant factors in the evolution of our species (Homo sapiens). As humans dispersed from Africa around 100 kya, marine resources provided essential nutrients in island environments of the tropical Southeast Asian seas. The Philippine archipelago has revealed significant evidence of early human settlement during this period of coastal migration. Yet, despite being a global marine biodiversity hotspot, few studies have investigated prehistoric marine adaptations there. In this study, we focus on the analysis of fish bones from three sites in Occidental Mindoro, Philippines: Bubog I, Bubog II, and Bilat Cave with the aim of enhancing the understanding of H. sapiens' coastal adaptations and maritime interactions since at least 32,000 BP. We utilized expanded skeletal element identification protocols to determine the lowest taxonomic level possible, combined with diversity indices. The analyses indicate that H. sapiens on Mindoro mostly exploited near-shore environments, with temporal variations in fishing practices reflecting differences in site occupation intensity and periods of climate-mediated habitat change. Inter-site variations in fishing practices are likely related to disparities in local marine habitats. Spatiotemporal differences in marine environments influenced the development of diverse fishing methods and technologies, demonstrating a detailed knowledge of local aquatic environments and the adaptability of modern humans in Island Southeast Asia since the Pleistocene.

Unknown Tag Identification Protocol Based on Collision Slot Resolution in Large-Scale and Battery-Less RFID System

Radio frequency identification (RFID) technology is widely used in warehouse management and supply chain management. In the actual application, unknown tags such as newly added or incorrectly placed products will be placed into a certain area of the warehouse. Existing identification protocols can identify such unknown tags by collecting tag IDs and comparing them with back-end server. Alternatively, the known tags can be filtered through the filter vector to prevent interference from unknown tags and improve identification efficiency. However, there is a lack of consideration of the collision impact on identification efficiency. In this paper, we introduce the bit tracking technology to accelerate the identification process and propose a Bit-Tracking Based Strategy for Unknown Tag Identification (BTUTI) protocol. We solve the problem by means of known tag deactivation and unknown tag identify. First, by adopting bit tracking technology, the reader can easily locate the positions of the collision bit. By hashing the tags of these collision slots, the collision slots can be turned into resolvable slots, thus improving the utilization rate of slots. Second, in the expected non-empty slot, the reader can send command to deactivate the known tag and skip empty slots and irresolvable collision slots. The experiment proves that BTUTI can increase the time efficiency compared to the state-of-the-art.

55 Dysphagia identification practices in residential long-term care settings in the Republic of Ireland: a cross-sectional electronic survey

Abstract Background Dysphagia is common in older adults with neurological and/or neurodegenerative conditions requiring early identification to avoid serious complications, including malnutrition, dehydration and aspiration pneumonia. The aim of this study was to describe dysphagia identification practices in Residential Long-Term Care Settings (RLTCS) in the Republic of Ireland. Methods A cross sectional electronic survey was designed using Qualtrics XM Platform that included 40 questions divided in 5 sections. The survey was circulated via email to RLTCS to be completed by either a clinical nurse or nurse manager in each facility. Descriptive data analysis was conducted on survey responses using RStudio. Results There were 45 surveys completed out of N = 429 electronically circulated (10% response rate). All RLTCS that completed the survey reported to routinely implement dysphagia identification practices. Observation of signs and symptoms of dysphagia at mealtimes (100%), review of clinical records to identify dysphagia risk factors and comorbidities (93%) and resident self-report of swallowing difficulties (91%) were the most common dysphagia identification practices. Poor oral motor function (89%) and history of aspiration pneumonia (87%) were reported as the main dysphagia risk factors. A swallow screening test was less commonly used to identify dysphagia (42%). Coughing or choking on food or fluids, inability to swallow medication, oral or pharyngeal residue, delayed or absent initiation of swallow, leaking of food or fluids and difficulty with coordination of breathing and swallowing were identified by all respondents as key criteria for risk of dysphagia. Almost all participants reported that residents at risk of dysphagia were routinely referred to speech and language therapy services for further assessment. Conclusion A range of practices were identified in RLTCS that participated in the survey. Further research approaches are required to provide a more in-depth perspective of dysphagia identification practices and protocols used in RLTCS for timely and accurate identification of dysphagia to avoid associated complications.

Enhanced therapeutic effects of mesenchymal stem cell-derived extracellular vesicles within chitosan hydrogel in the treatment of diabetic foot ulcers

Extracellular vesicles (EVs) derived from human umbilical cord mesenchymal stem cells (hUCMSCs) have emerged as promising candidates for cell-free therapy in various diseases, including chronic cutaneous wounds. However, the lack of standardized protocols for EVs’ preparation and identification poses a significant challenge to their clinical application. Thus, the objective was to develop a safe and efficient method for the large-scale production of hUCMSC-derived EVs while establishing a comprehensive identification protocol encompassing morphology, particle size distribution, protein expression, and purity. This study observed that most of the EVs acquired through the protocol exhibited either a cup-shaped or round-shaped structure, with a median diameter of ~73.25 nm. The proportions of EVs positive for CD9, CD63, and CD81 were 37.5%, 38.6%, and 19.8%, respectively. To enhance their therapeutic potential in wound treatment, EVs were incorporated into chitosan hydrogel, forming chitosan hydrogel-EVs (CS-EVs). Furthermore, it was demonstrated that CS-EVs exhibited continuous release of EVs into the surrounding environment and, importantly, that the released EVs were internalized by human umbilical vein endothelial cells (HUVECs), resulting in significant enhancement of cell migration and angiogenesis. Additionally, in a rat model of diabetic foot ulcers, CS-EVs demonstrated a robust therapeutic effect in promoting wound healing. Following a 15-day treatment period, the group treated with CS-EVs demonstrated an impressive 93.3% wound closure ability, accompanied by a high degree of re-epithelialization. In contrast, the control group exhibited only a 71.5% reduction in wound size. In summary, this study offers solutions for the purification, characterization, and application of EVs in clinical wound treatment. These results not only offer fresh perspectives on the involvement of hUCMSC-derived EVs in wound healing but also introduce a non-invasive approach for applying EVs that holds practical significance in skin repair.Graphical

Femoroacetabular Impingement in Ice Hockey Athletes.

Femoroacetabular impingement (FAI) in ice hockey is a concern for many athletes. The biomechanics of skating and the injury mechanism, prevalence, identification, and treatment protocols currently available for FAI in ice hockey athletes are important for all coaches and practitioners to understand. This article discusses the underlying anatomical issues and biomechanical considerations surrounding FAI. Furthermore, this article describes the interventions that can be used when encountering FAI and well-established protocols to aid in the return to play. Finally, prevention strategies that can aid in injury prevention are discussed.

Structure identification and inhibitory mechanism evaluation of three novel angiotensin converting enzyme (ACE) inhibitory peptides from small-aroma chicken

Small-aroma chicken with a high protein content, is widely distributed in the mountainous area of Yunnan-Guizhou Plateau, China. In this work, three novel ACE-inhibitory peptides, Leu-Thr-Glu-Lys-Val-Val-Phe (LTQKVVF), Leu-Asp-Asp-His-Phe-Leu (LDDHFL), and Val-Pro-Gly-Pro-Glu-Pro-Lys-Pro (VPGPEPKP), were prepared from the Small-Aroma Chicken breast. The purification and identification protocol employed comprised ion-exchange chromatography, gel filtration chromatography, reverse phase-high performance liquid chromatography (RP-HPLC), and nano liquid chromatography-electrospray ionization–tandem mass (Nano-LC-ESI–MS/MS). Compared with VPGPEPKP and VPGPEPKP, LTQKVVF displayed noticeable ACE inhibitory activity, with IC50 values of 189.34 μM. What’s more, the Lineweaver-Burk plots revealed peptides LTQKVVF and LDDHFL acted as the mixed-competitive inhibitor while VPGPEPKP inhibited ACE in a manner similar to uncompetitive inhibition. The binding free energies of LTQKVVF, LDDHFL, and VPGPEPKP for ACE were −8.1 kcal/mol, −8.6 kcal/mol, and −9.6 kcal/mol, respectively. The strong inhibition of ACE by VPGPEPKP may be attributed to the interaction of hydrogen bonds, alkyl, and pi-alkyl, which formed by crucial residues of ACE.

Efficient Privacy-Preserving Face Identification Protocol

Face identification is an important research topic in artificial intelligence and it has wide application in civilian and law enforcement scenarios that require reliable biometric identification. However, the wide deployment of face identification systems has raised serious privacy concerns about the abuse of users' face images. In this paper, we propose an efficient privacy-preserving face identification protocol based on fully homomorphic encryption and garbled circuit. Our work is the first privacy-preserving face identification protocol that guarantees the privacy of the users' face images such that the server cannot learn any information about the face images, access pattern and identification result, while providing face identification service. We formally prove the security of our protocol in a semi-honest model. Experimental results the practicality of our protocol for the large face images database.

Antimicrobial and antioxidant potentials of an endophytic cunninghamella sp. isolated from the leaves of Chrysophyllum albidum

Background: Several studies have identified endophytic fungi associated with Nigerian plants as potential sources of new drug discovery. Aim: This study was carried out to investigate the antimicrobial and antioxidant properties of secondary metabolites isolated from an endophytic Cunninghamella sp. associated with leaves of Chrysophyllum albidum, a Nigerian indigenous plant. Method: The plant material was collected in Agulu, Anambra State, South-East Nigeria. Endophytic fungal isolation and identification, as well as fermentation and extraction of fungal secondary metabolites, were carried out using standard methods. The antimicrobial and antioxidant activities of the fungal extract were evaluated using the agar well diffusion method and the 1,1-diphenyl-2-picryl-hydraxyl (DPPH) free radical assay, respectively. Result: An endophytic fungus was isolated from the leaves of C. albidum. Following a molecular identification protocol of DNA amplification and sequencing of the fungal ITS region, the fungus was identified as Cunninghamella sp. The fungal extract exhibited antimicrobial activity against the test bacteria and fungus. At concentrations ranging from 2.5 - 20 mg/mL, the extract produced inhibition zone diameters (IZD) ranging from 2 - 11 mm against Pseudomonas aeruginosa. Antibacterial activity was also observed against Staphylococcus aureus at concentrations ranging from 10 - 20 mg/mL, with IZD ranging from 3 - 6 mm. Antifungal activity was observed against Candida albicans at concentrations ranging from 1.25 - 20 mg/mL, with IZD ranging from 2 - 7 mm. The fungal extract demonstrated significant antioxidant activity in the DPPH assay when compared to the standard control, quercetin. The extract showed a percentage inhibition range of 95 - 98% at concentrations ranging from 20 to 100 µg/mL. Conclusion: The findings of this study highlight the potential of endophytic Cunninghamella sp. and other endophytic microorganisms of indigenous Nigerian plants as sources of bioactive compounds with antimicrobial and antioxidant properties for pharmaceutical and industrial applications.

MALDI-TOF MS-Based KPC Direct Detection from Patients' Positive Blood Culture Bottles, Short-Term Cultures, and Colonies at the Hospital.

Carbapenemase resistance in Enterobacterales is a global public health problem and rapid and effective methods for detecting these resistance mechanisms are needed urgently. Our aim was to evaluate the performance of a MALDI-TOF MS-based "Klebsiella pneumoniae carbapenemase" (KPC) detection protocol from patients' positive blood cultures, short-term cultures, and colonies in healthcare settings. Bacterial identification and KPC detection were achieved after protein extraction with organic solvents and target spot loading with suitable organic matrices. The confirmation of KPC production was performed using susceptibility tests and blaKPC amplification using PCR and sequencing. The KPC direct detection (KPC peak at approximately 28.681 Da) from patients' positive blood cultures, short-term cultures, and colonies, once bacterial identification was achieved, showed an overall sensibility and specificity of 100% (CI95: [95%, 100%] and CI95: [99%, 100%], respectively). The concordance between hospital routine bacterial identification protocol and identification using this new methodology from the same extract used for KPC detection was ≥92%. This study represents the pioneering effort to directly detect KPC using MALDI-TOF MS technology, conducted on patient-derived samples obtained from hospitals for validation purposes, in a multi-resistance global context that requires concrete actions to preserve the available therapeutic options and reduce the spread of antibiotic resistance markers.

Impact of elastic modulus evolution on springback prediction

Today, the hardening Yoshida-Uemori model is implemented in the most used FEA stamping codes (Autoform®, Pamstamp®, LS-Dyna®…) with occasionally some variations. This forming material card is requested regularly by the car manufacturers. The Yoshida-Uemori model is a combined hardening model coupled with the elastic evolution modulus which depends on the equivalent plastic strain. According to the experimental tests conducted and their identification protocol, the set of parameters improves the springback prediction compared to the prediction with a purely isotropic hardening model. The numerical analysis shows that the elastic modulus evolution has an influence on springback prediction of the same order as the combined hardening (isotropic and kinematic). The adjusting parameters governing the evolution of the elastic modulus are identified thanks to hysteresis loops conducted between 0.2% of plastic deformation and uniform elongation (Uel). For each loop, there are several ways to define an unloading elastic module: the chord of loop; the slope deducted from the linear regression on unloading path on a range. According to the choice of range, the value differs. What is the impact of these choices on the springback prediction of a stamped part?

Focus Issue Articles on Diagnostic Assay Development and Validation: The Science of Getting It Right

Focus Issue Articles on Diagnostic Assay Development and Validation: The Science of Getting It Right

How to verify identity in the continuous variable quantum system?

Continuous variable quantum cryptography has developed rapidly in recent decades, but how to verify identity in the continuous variable quantum system is still an urgent issue. To solve this problem, we propose a continuous variable quantum identification (CV-QI) protocol based on the correlation of the two-mode squeezed vacuum state and the continuous variable teleportation. The bidirectional identity verification between two participants of the communication can be achieved by the proposed CV-QI protocol. To guarantee the security, we make full use of the decoy state sequences during the whole process of the proposed CV-QI protocol. Besides, we provide the security analyses of the proposed CV-QI protocol, and analyses indicate that the security of the proposed CV-QI protocol is guaranteed.

Study of knowledge, attitude, and practices of child care physicians toward nutritional vitamin D deficiency in pediatric population

Objectives: Nutritional Vitamin D deficiency (NVDD), commonly seen in infancy and childhood, is a nutritional epidemic. Practicing child care physicians have different approaches to the diagnosis and treatment of nutritional Vitamin D deficiency in children. This study aims to assess the knowledge, attitude, and practice of clinical diagnosis, investigations, and treatment of NVDD in infants and children among child care physicians from a teaching institute. Material and Methods: A cross-sectional study was conducted to assess knowledge, attitude, and practice of NVDD among resident doctors, consultants and faculty members working in a teaching hospital by convenience sampling method. The study was based on a pretested questionnaire which was administered by a single investigator on all the study participants. Twelve questions formulated to assess the knowledge, was scored as 0 (0%) to 14(100%) and ten questions regarding attitude and practices with possible responses as agree, disagree, or neutral was scored as 0 (0%) to 20 (100%). Data were analysed with statistical software, STATA, version 10.1,2011. Descriptive statistics included summary measures such as frequency and percentages to summarize qualitative variables and categorical responses. Inferential statistics included P-values generated from hypothesis testing procedures. Results: There were 96 participants including 46 residents. Forty-seven (49%) of them scored more than 11 out of 14 while 49 (51%) participants scored between 7 to 10 out of 14 on the knowledge part of the questionnaire. Interestingly nobody scored less than 7 out of 14. Sixty (62%) participants scored more than 15 out of 20 and nearly 19 (2%) physicians scored less than 10 out of 20 in the attitude and practice part of the questionnaire. On intergroup comparison between consultant pediatricians and residents on knowledge regarding NVDD showed no significant difference except for knowledge regarding cumulative therapeutic dose of cholecalciferol (p value 0.025). There were more attitude and practice gaps in this group of respondents although it was not statistically significant. Conclusion: This study identifies gaps in the knowledge, attitude and practice of child care physicians about NVDD in infants and children. Knowledge gaps regarding definition and change in treatment protocol of NVDD and inconsistency in clinical identification, diagnosis, treatment, and prevention protocols across all groups of physicians.

Quantitative Detection of Microplastics in Water through Fluorescence Signal Analysis

Microplastics (MPs) have recently been acknowledged as a new major and ubiquitous environmental pollutant with still unclear, yet potentially high, risks for different ecosystems and human health. Nevertheless, quantitative identification protocols rely on long and subjective visual counting necessarily performed on microscopes by well-trained operators. In this study, an automatic, fast, portable, and inexpensive method for the quantitative detection of MPs in water is proposed. The system is based on the typical optical setup of a fluorescence confocal microscope specifically adapted to automatically count dye-stained MPs in flowing liquids using a low-power laser beam. The fluorescence pulses emitted by flowing MPs are revealed and processed by a specific software using a pattern recognition algorithm to discriminate and count real fluorescence pulses out of noise fluctuations. The system was calibrated with commercial orange fluorescent 10 µm and 1 µm polystyrene microspheres, and remarkable agreement with theoretical predictions was obtained regarding different parameters. Tests were also performed with laboratory-prepared MPs dispersed in different types of real water samples. In this case, the agreement with theory was slightly worse and differences found in the quantitative results require further investigation. However, the present study demonstrated the proof of concept of a method for quick automated MP counting in water.

The confidence-accuracy relationship for lineup decisions holds for the Dutch identification procedure.

Post-decision confidence from witnesses who make a positive identification decision can serve as a valuable indicator of identification accuracy under certain conditions. International best-practice guidelines therefore recommend asking witnesses to indicate their confidence following a selection from a lineup. Three experiments that used Dutch identification protocols, however, reported no significant post-decision confidence-accuracy association. To examine this conflict between the international and the Dutch literature, we tested the strength of the post-decision confidence-accuracy relationship for lineups that followed Dutch protocol in two ways: we conducted an experiment and re-analyzed two experiments that implemented Dutch lineup protocols. As expected, the post-decision confidence-accuracy relationship was strong for positive identifications and weak for negative identification decisions in our experiment. The re-analysis of the pre-existing data showed a strong effect for positive identification decisions of participants up to the age of 40 years. For exploratory purposes, we also tested the confidence-accuracy relationship between lineup administrators' perception of witnesses' confidence and eyewitness identification accuracy. In our experiment, the relationship was strong for choosers and weak for nonchoosers. The re-analysis of pre-existing data showed no correlation between confidence and accuracy, unless we excluded adults over 40 of age. We recommend adapting the Dutch identification guidelines to reflect the current and previous findings on the post-decision confidence-accuracy relationship.

The establishment of transient expression systems and their application for gene function analysis of flavonoid biosynthesis in Carthamus tinctorius L

BackgroundSafflower (Carthamus tinctorius L.) is an important economic crop and a traditional medicinal material rich in flavonoids, which can alleviate cardiovascular and cerebrovascular pathologies. Thus, many candidate genes involved in safflower flavonoid biosynthesis have been cloned. However, owing to the lack of a homologous gene expression system, research on gene function is limited to model plants. Therefore, a gene function identification protocol for safflower must be established.ResultsIn the present study, using safflower callus as the experimental material, Agrobacterium and biolistic transient expression systems were established. In the Agrobacterium transient expression system, the highest transformation rate was obtained at the original Agrobacterium concentration of OD600 0.4, infiltration concentration of OD600 0.6, infection for 20 min, co-culture for 3 days, and acetosyringone concentration of 100 μmol·L−1. In the biolistic transient expression system, the highest transformation efficiency was observed at helium pressure of 1,350 psi, vacuum degree of -0.8 bar, flight distance of 6.5 cm, one round of bombardment, plasmid concentration of 3 μg·shot−1, and gold particle concentration of 100 μg·shot−1. Further, these two transient expression systems were used for the functional analysis of CtCHS1 as an example. After overexpression, relative CtCHS1 expression increased, particularly in Agrobacterium-transformed calli. Additionally, the contents of some flavonoids were altered; for instance, naringenin and genistein levels were significantly increased in Agrobacterium-transformed calli, whereas luteolin, luteolin-7-O-rutinoside, and apigenin derivative levels were significantly decreased in biolistic-transformed calli.ConclusionUsing safflower callus as the experimental material, highly efficient Agrobacterium and biolistic transient expression systems were successfully established, and the utility of both systems for investigating gene function was demonstrated. The proposed safflower callus transient expression systems will be useful for further functional analyses of flavonoid biosynthetic genes in safflower.