Faba Bean Processing: Thermal and Non-Thermal Processing on Chemical, Antinutritional Factors, and Pharmacological Properties.

The food industry, academia, food technologists, and consumers have become more interested in using faba bean seeds in the formulation of new products because of their nutritional content, accessibility, low costs, environmental advantages, and beneficial impacts on health. In this review, a systematic and up-to-date report on faba bean seeds' antinutrients and bioactive and processing techniques is comprehensively presented. The chemical composition, including the oil composition and carbohydrate constituents, is discussed. Factors influencing the reduction of antinutrients and improvement of bioactive compounds, including processing techniques, are discussed. Thermal treatments (cooking, autoclaving, extrusion, microwaving, high-pressure processing, irradiation) and non-thermal treatments (soaking, germination, extraction, fermentation, and enzymatic treatment) are identified as methods to reduce the levels of antinutrients in faba bean seeds. Appropriate processing methods can reduce the antinutritional factors and enrich the bioactive components, which is useful for the seeds' efficient utilization in developing functional foods. As a result, this evaluation focuses on the technologies that are employed to reduce the amounts of toxins in faba bean seeds. Additionally, a comparison of these methods is performed in terms of their advantages, disadvantages, viability, pharmacological activity, and potential for improvement using emerging technologies. Future research is expected in this area to fill the knowledge gap in exploiting the nutritional and health benefits of faba bean seeds and increase the utilization of faba bean seeds for different applications.

Statistical Modelling Investigation of MALDI-MSI-Based Approaches for Document Examination.

Questioned document examination aims to assess if a document of interest has been forged. Spectroscopy-based methods are the gold standard for this type of evaluation. In the past 15 years, Matrix-Assisted Laser Desorption Ionisation-Mass Spectrometry Imaging (MALDI-MSI) has emerged as a powerful analytical tool for the examination of finger marks, blood, and hair. Therefore, this study intended to explore the possibility of expanding the forensic versatility of this technique through its application to questioned documents. Specifically, a combination of MALDI-MSI and chemometric approaches was investigated for the differentiation of seven gel pens, through their ink composition, over 44 days to assess: (i) the ability of MALDI MSI to detect and image ink chemical composition and (ii) the robustness of the combined approach for the classification of different pens over time. The training data were modelled using elastic net logistic regression to obtain probabilities for each pen class and assess the time effect on the ink. This strategy led the classification model to yield predictions matching the ground truth. This model was validated using signatures generated by different pens (blind to the analyst), yielding a 100% accuracy in machine learning cross-validation. These data indicate that the coupling of MALDI-MSI with machine learning was robust for ink discrimination within the dataset and conditions investigated, which justifies further studies, including that of confounders such as paper brands and environmental factors.

Quantification of proteins in whole blood, plasma and DBS, with element-labelled antibody detection by ICP-MS.

Over recent years, quantification of multiple proteins in body fluids has become increasingly prominent, which is beneficial to a number of scientific fields, not least biomedical. Several techniques have been developed based on conventional ELISA; one of these techniques is analysis of proteins labelled with element-tagged antibodies by ICP-MS in serum, allowing quantification of multiple targets within a single sample. This research aimed to quantify albumin and immunoglobulin G (IgG) levels in plasma, whole blood and dried blood spots using NANOGOLD® and Europium labelled antibodies analysed by ICP-MS. Before the proteins were quantified simultaneously, albumin and IgG concentrations were measured separately and compared to protein levels obtained by ELISA. It was found that protein concentrations for both albumin and IgG obtained with element-labelled antibody detection correspond to those determined by ELISA. Furthermore, albumin and IgG levels measured simultaneously by ICP-MS correspond to concentrations found when the proteins were analysed separately by ICP-MS. Finally, development of this method has provided a positive indication that it can be extended to quantification of additional proteins, which could be related to a disease or as a minimum provide additional information for a protein profile of an individual.

Rapid detection of cocaine, benzoylecgonine and methylecgonine in fingerprints using surface mass spectrometry.

Latent fingerprints provide a potential route to the secure, high throughput and non-invasive detection of drugs of abuse. In this study we show for the first time that the excreted metabolites of drugs of abuse can be detected in fingerprints using ambient mass spectrometry. Fingerprints and oral fluid were taken from patients attending a drug and alcohol treatment service. Gas chromatography mass spectrometry (GC-MS) was used to test the oral fluid of patients for the presence of cocaine and benzoylecgonine. The corresponding fingerprints were analysed using Desorption Electrospray Ionization (DESI) which operates under ambient conditions and Ion Mobility Tandem Mass Spectrometry Matrix Assisted Laser Desorption Ionization (MALDI-IMS-MS/MS) and Secondary Ion Mass Spectrometry (SIMS). The detection of cocaine, benzoylecgonine (BZE) and methylecgonine (EME) in latent fingerprints using both DESI and MALDI showed good correlation with oral fluid testing. The sensitivity of SIMS was found to be insufficient for this application. These results provide exciting opportunities for the use of fingerprints as a new sampling medium for secure, non-invasive drug detection. The mass spectrometry techniques used here offer a high level of selectivity and consume only a small area of a single fingerprint, allowing repeat and high throughput analyses of a single sample.

Rapid Operationalization of COVID-19 Immunization Clinics With Medical and Physician Assistant Students Serving as Vaccinators.

Introduction: In anticipation of institutional and community-wide COVID-19 immunization clinics, an educational program for the administration of COVID-19 vaccines was developed, collaborating with the Virginia Department of Health to train doctor of medicine and physician assistant students to serve as vaccinators. Faculty and students also worked with state legislatures to propose and enact a bill that would enable such students to vaccinate patients. Methods: Between January 2021 and August 2022, 263 student volunteers completed 3,685 person-hours, administering 48,279 doses. On the basis of community need, the majority of vaccines were administered at mass vaccination clinics in Chesapeake (47%) and Norfolk (22%) in Virginia. One year after the first COVID-19 immunization clinic, the authors surveyed students who assisted with clinics, utilizing a Likert scale and free-text responses to elicit feedback about the training and volunteer experience. Results: Volunteers ranked the vaccination clinics among the top third of established volunteer experiences offered at Eastern Virginia Medical School, and 75% of respondents believed that this training should be a permanent part of their program curriculum. Conclusions: This paper presents a strategy for preparing students to serve as vaccinators during public health emergencies and show how other institutions of medical education can prepare for and engage student participation in vaccination campaigns and emergent health initiatives.

Spectroscopic imaging based approach for condom identification in condom contaminated fingermarks.

Sexual offenders are increasingly reported to use condoms while committing the crime, mainly to prevent the transfer of DNA evidence. Although condoms are often removed from the crime scene, vaginal swabs can be taken from the victim to prove the presence of condom lubricants and therefore evidence of corpus delicti. However, late reporting to the police and the tendency of the victim to wash immediately after the crime, may compromise the detection of condom lubricants. Recently we showed that Matrix-Assisted Laser Desorption/Ionisation MS Imaging (MALDI MSI) of condom contaminated fingermarks enables images of the fingermark ridge pattern to be obtained simultaneously with the detection of the condom lubricant for two condom brands, thus becoming a potential alternative way to link the assailant to the crime. Building on the value of this information, it would be advantageous to identify the condom brand used during the sexual assault. Here we show the development of a multidisciplinary spectroscopic approach, including MALDI MSI, MS/MS, Raman microscopy and ATR-FTIR spectroscopy, applied to a range of condom brands/types. The techniques have complementary features and provide complementary information to retrieve a "condom brand spectroscopic fingerprint". Unique spectroscopic profiles would greatly aid in the screening and identification of the condom, thus adding intelligence to the case under investigation.

MALDI MS Imaging of Cucumbers.

There are many different methodologies in the literature for the preparation of plant material for subsequent MALDI MSI analysis. This chapter overviews preparation of cucumbers (Cucumis sativus L.), with emphasis on sample freezing, cryosectioning, and matrix deposition. This should act as a representative example of sample preparation for plant tissue, and due to wide sample variation (e.g., leaves, seeds, and fruit) and analytes of interest, method optimization will be required for different samples.

Faba Bean Flavor Effects from Processing to Consumer Acceptability.

Faba beans as an alternative source of protein have received significant attention from consumers and the food industry. Flavor represents a major driving force that hinders the utilization faba beans in various products due to off-flavor. Off-flavors are produced from degradation of amino acids and unsaturated fatty acids during seed development and post-harvest processing stages (storage, dehulling, thermal treatment, and protein extraction). In this review, we discuss the current state of knowledge on the aroma of faba bean ingredients and various aspects, such as cultivar, processing, and product formulation that influence flavour. Germination, fermentation, and pH modulation were identified as promising methods to improve overall flavor and bitter compounds. The probable pathway in controlling off-flavor evolution during processing has also been discussed to provide efficient strategies to limit their impact and to encourage the use of faba bean ingredients in healthy food design.

Detection of a novel allele, HLA-DPA1*01:58, originating from a recombination within exon 2 of the DPA1 gene.

A crossover in exon 2 between DPA1*01:03:01:02 and DPA1*02:01:01:01 results in a novel hybrid allele, HLA-DPA1*01:58.

The novel allele HLA-DQA1*05:05:10 was detected in a Nigerian who also lacks the DRB5 gene in the DRB1*16 haplotype.

One synonymous nucleotide substitution in exon 2 of DQA1*05:05:01:13 results in a novel allele, HLA-DQA1*05:05:10.

A novel matrix-assisted laser desorption/ionisation mass spectrometry imaging based methodology for the identification of sexual assault suspects.

An increase in the use of condoms by sexual offenders has been observed. This is likely to be due both to the risk of sexually transmitted diseases and to prevent the transfer of DNA evidence. In this scenario the detection of condom lubricants at a crime scene could aid in proving corpus delicti. Here we show a novel application of Matrix-Assisted Laser Desorption/Ionisation Mass Spectrometry Imaging (MALDI MSI) for mapping the fingermark ridge pattern simultaneously to the detection of the condom lubricant within the fingermark itself. Two condom brands have been investigated to prove the concept. Condoms were handled producing lubricant-contaminated fingermarks. Images of the ridge pattern were obtained simultaneously to the detection of two lubricants, even several weeks after the fingermark deposition. The results therefore show the potential of MALDI MSI to link the suspect (identification through fingermark ridge pattern) to the crime (detection of condom lubricant) in one analysis. This would enable forensic scientists to provide evidence with stronger support in alleged cases of sexual assault.

Patient Interest in Video Integration for After-Hours Telemedicine.

PURPOSE: To understand patient attitudes, access toward video calling to enhance efficiency of after-hours triage calls. METHODS: We surveyed patients aged 18 to 89 years. Questions included demographics, preferences, access to video calling devices, and perceived advantages and disadvantages of this technology. Answers were entered into Qualtrics database and analyzed using JMP 11 (SAS, Cary, NC). RESULTS: Two hundred ninety-eight patients agreed to participate. Mean age was 47.9 years; 71.6% were female; and 75.1% had access to video calling device. Device proficiency was inversely related to age and greatest in 18-to-32-years group (χ2 = 71.18, P < .0001). Seventy-one percent of patients enjoyed video communication, directly proportional to education (trend test Z = 2.78, P < .005). Adjusted for both age and education, respondents with college education or above were 3 times more likely to self identify as "good' with video (OR, 3.11; 95% CI, 1.48-6.64); those under age 48 had even higher proficiency (Odds ratio (OR), 13.9; 95% CI, 4.79-59.34). Patients with prior video experience were 3 times more likely to prefer video calling (Relative risk (RR) = 3.46; 95% CI, 1.95-6.11). Patients calling their doctor 5 or more times annually preferred video calling significantly more than calling by telephone (RR, 1.61; 95% CI, 1.31-1.97). Faster contact with the primary care provider (19.8%) was the most perceived advantage. Loss of in-person interaction with doctor (37.1%) was the greatest perceived disadvantage. CONCLUSIONS: Patients seem to have access and interest in video communication for after-hours calls. Further studies are needed to evaluate whether addition of video component to after-hours triage calls will help reduce unnecessary emergency department visits.

Study of latent fingermarks by matrix-assisted laser desorption/ionisation mass spectrometry imaging of endogenous lipids.

Identification of suspects via fingermark analysis is one of the mainstays of forensic science. The success in matching fingermarks, using conventional fingermark scanning and database searching, strongly relies on the enhancement method adopted for fingermark recovery; this in turn depends on the components present in the fingermarks, which will change over time. This work aims to develop a robust methodology for improved analytical detection of the fingermark components. For the first time, matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) has been used to image endogenous lipids from fresh and aged, groomed and ungroomed fingermarks. The methodology was initially developed using oleic acid which was detected along with its degradation products over a 7-day period, at three different temperatures in a time-course experiment. The optimised methodology was then transferred to the imaging analysis of real fingermark samples. Fingermark patterns were reconstructed by retrieving the m/z values of oleic acid and its degradation products. This allowed the three aged fingermarks to be distinguished. In order to prove that MALDI-MSI can be used in a non-destructive way, a simple washing protocol was adopted which returned a fingermark that could be further investigated with classical forensic approaches. The work reported here proves the potential and the feasibility of MALDI-MSI for the forensic analysis of fingermarks, thus making it competitive with other MSI techniques such as desorption electrospray ionisation (DESI)-MS. The feasibility of using MALDI-MSI in fingermark ageing studies is also demonstrated along with the potential to be integrated into routine fingermark forensic analysis.

MALDI Mass Spectrometry Profiling and Imaging Applied to the Analysis of Latent Fingermarks.

Latent fingermarks are derived from a transfer of material from the fingertips to a surface upon contact. Traditionally, fingermarks are employed for biometric identification of individuals based on matching of the pattern of the ridges. However, in recent years, there has been a stark increase in the use of advanced analytical techniques in order to obtain additional information, specifically the chemical composition of the residue. Understanding the complexity of the endogenous and exogenous content of fingermarks could be extremely useful in allowing further development of enhancement techniques currently used in forensic scenarios by identifying potential target molecules. This chemical information could also potentially provide invaluable information on the lifestyle of an individual, including their activities prior to depositing a mark.An analytical tool that has gained notable popularity in this novel area of research is matrix-assisted laser desorption/ionisation mass spectrometry (MALDI MS). This technique can either be employed for rapid chemical profiling or imaging of fingermarks to detect chemical species contained within the residue, with the latter also allowing for physical reconstruction of the fingermark ridges.This chapter will provide an overview of the protocols employed to allow for both MALDI MS profiling and imaging analysis of latent fingermarks, specifically covering the types of fingermarks employed and techniques used to deposit matrices onto samples.

Unambiguous high resolution genotyping of human leukocyte antigens.

We have developed a high resolution sequencing based typing method for genotyping Human Leukocyte Antigens (HLA) over a period of twenty years. The methods are based upon the separation of HLA alleles per locus at the initial amplification to simplify the analysis post-sequencing. The increasing discovery of polymorphism in HLA, manifested in new alleles, has necessitated the continuing development of this method. Here we present methods for the high resolution Sequence Based Typing of HLA-A, B, C (class I) and HLA-DQB1 and DRB1 (class II). The purpose of this article is to provide a valuable resource of methods and primers for other laboratories engaged in HLA typing.

MALDI-MS imaging for the study of tissue pharmacodynamics and toxicodynamics.

Pharmacodynamics and toxicodynamics are the study of the biochemical and physiological effects of therapeutic agents and toxicants and their mechanisms of action. MALDI-MS imaging offers great potential for the study of pharmaco/toxicodynamic responses in tissue owing is its ability to study multiple biomarkers simultaneously in a label-free manner. Here, existing examples of such studies examining anticancer drugs and topically applied treatments are described. Examination of the literature shows that the use of MS imaging in pharmaco/toxicodynamic studies is in fact quite low. The reasons for this are discussed and potential developments in the methodology that might lead to its further use are described.

Towards the integration of matrix assisted laser desorption ionisation mass spectrometry imaging into the current fingermark examination workflow.

A wide range of fingermark enhancement techniques (FET) is currently employed to visualise latent fingermarks at crime scenes. However, if smudged, partial, distorted or absent in the National Fingerprint Database, crime scene marks may be not useful for identification purposes. In these circumstances, a technology enabling chemical imaging of both endogenous and exogenous species contained within the fingermark could provide additional and associative investigative information, to profile the suspect's activities prior to the crime. Matrix Assisted Laser Desorption Ionisation Mass Spectrometry Imaging (MALDI MSI) has proven to be such a technique, enabling investigative information to be gathered, for example, on what substances the donor has come in contact with and what they have ingested. Nonetheless, to be employed, MALDI MSI has to be validated and its compatibility with FET tested for integration into the standard fingermark examination workflow. For the first time, a direct comparison has been made between the efficiency of a range of FET and MALDI MSI under different conditions. This information will build towards validation of the technology. Also, for the first time, MALDI MSI has been successfully employed as a sequential step following fingermark enhancement using many of the currently employed FET. Additionally, known enhancers have been "re-visited" by combining them with a MALDI matrix, providing both improved fingermark development and chemical species detection via MALDI MSI. The result reported here are good indication in favour of the integration of MALDI MSI into the current fingermark examination workflow for gathering additional investigative information.

Seasonal cycles in whole-body proximate composition and energy content of forage fish vary with water depth.

Quantifying the nutritional quality of forage fish is integral for understanding upper trophic levels as forage fish are the dominant prey for top predator fish, marine mammals, and sea birds. Many existing reports documenting body composition of forage species are not comparable due to confounding effects. This study systematically assessed the variability in proximate composition and energy content of 16 forage species in southeastern Alaska (57.2626 N/133.7394 W) between 2001 and 2004. Variation in energy and lipid contents was related to habitat, epipelagic planktivores varying most, mesopelagics intermediate, and demersal species relatively invariable. Season was the greatest source of variation as a result of short growing seasons at high latitude and energy allocation strategies for reproduction and growth. Among species that varied seasonally, energy and lipid increased over summer and declined during winter. Annual differences in body composition occurred during periods of peak energy content. Sampling recommendations and guidance for bioenergetics models are provided.

Yellow fever vaccine: an updated assessment of advanced age as a risk factor for serious adverse events.

Since 1996, the scientific community has become aware of 14 reports of yellow fever vaccine (YEL)-associated viscerotropic disease (YEL-AVD) cases and four reports of YEL-associated neurotropic disease (YEL-AND) worldwide, changing our understanding of the risks of the vaccine. Based on 722 adverse event reports after YEL submitted to the U.S. Vaccine Adverse Event Reporting System in 1990-2002, we updated the estimates of the age-adjusted reporting rates of serious adverse events, YEL-AVD and YEL-AND. We found that the reporting rates of serious adverse events were significantly higher among vaccinees aged > or =60 years than among those 19-29 years of age (reporting rate ratio = 5.9, 95% CI 1.6-22.2). Yellow fever is a serious and potentially fatal disease. For elderly travelers, the risk for severe illness and death due to yellow fever infection should be balanced against the risk of a serious adverse event due to YEL.