RESUMO
RESEARCH QUESTION: What is the profile of women in the USA who become surrogates, and what is their power of decision and motivations? DESIGN: This quantitative study was performed with 231 participants in the USA, given the country's long history of surrogacy, to help clarify the profile of women who become surrogates, their power of decision and motivations. RESULTS: Descriptive and multivariate cluster analyses showed that women who become surrogates earn above the average income for their state of residency, have a high level of education, have health insurance, are employed, and decide to become a surrogate for prosocial/altruistic reasons. CONCLUSIONS: In contrast to the premise of both radical feminism and ultra-conservative Catholicism, this study found that altruism and empathy are the primary motivations for participating in surrogacy processes, and that a woman's decision to become a surrogate is not motivated by social conditioning relating to poverty or social status.
RESUMO
The use of ordnance gelatine has been widespread in the field of ballistics as a simulant for soft tissue when assessing ballistic threats. However, the traditional method of preparing ordnance gelatine is time-consuming and requires precision to ensure that the final mold meets the required specifications. Furthermore, temperature control is necessary post-production, and there are limitations on its usage duration. To address these issues, manufacturers have developed pre-mixed, gelatine-like products that are stable at room temperature and require less preparation time. Nonetheless, it is uncertain whether these new products can perform in the same manner as the gold standard of ordnance gelatine. This study used five types of blocks, including ordnance gelatine (10% and 20%), Clear Ballistics (10% and 20%) and Perma-Gel (10%) and subjected them to 9 mm, 0.380 Auto fired from a universal receiver and a 5.56 × 45 mm ammunition fired by a certified firearms instructor. Delta-V and total energy dissipation were measured after each test using data collected from ballistic chronographs placed in front of and behind each block. High-speed video was recorded, and a cut-down analysis conducted. The findings revealed variations in energy dissipation and fissure formation within the block, with greater energy based on fissure formation observed in the ordnance gelatine. Additionally, the high-speed video showed the occurrence of secondary combustions occurring in the premixed gelatines.
Assuntos
Balística Forense , Gelatina , Ferimentos por Arma de Fogo , Balística Forense/métodos , Humanos , Modelos BiológicosRESUMO
The protozoan parasites Trypanosoma brucei, Trypanosoma cruzi and Leishmania spp. are responsible for continued propagation of neglected tropical diseases such as African sleeping sickness, Chagas disease and leishmaniasis respectively. Following a report that captopril targets Leishmania donovani dipeptidyl carboxypeptidase, a series of simple proline amides and captopril analogues were synthesized and found to exhibit 1-2 µM in vitro inhibition and selectivity against Trypanosoma brucei, Trypanosoma cruzi and Leishmania spp. The results were corroborated with computational docking studies. Arguably, the synthetic proline amides represent the structurally simplest examples of in vitro pan antiprotozoal compounds.
Assuntos
Captopril , Trypanosoma brucei brucei , Trypanosoma cruzi , Captopril/farmacologia , Captopril/química , Captopril/síntese química , Trypanosoma brucei brucei/efeitos dos fármacos , Trypanosoma brucei brucei/enzimologia , Trypanosoma cruzi/efeitos dos fármacos , Trypanosoma cruzi/enzimologia , Relação Estrutura-Atividade , Simulação de Acoplamento Molecular , Tripanossomicidas/farmacologia , Tripanossomicidas/química , Tripanossomicidas/síntese química , Estrutura Molecular , Leishmania/efeitos dos fármacos , Leishmania/enzimologia , Antiprotozoários/farmacologia , Antiprotozoários/química , Antiprotozoários/síntese química , HumanosRESUMO
Optical surrogates, derived from absorbance and fluorescence spectra, are widely used to infer dissolved organic matter (DOM) composition (molecular weight, aromaticity) and genesis (autochthonous vs allochthonous). Despite the broad adoption of optical surrogates, several limitations exist, such as context- and sample-specific factors. These limitations create uncertainty about how compositional interpretations based on optical surrogates are generalized across contexts, specifically if there is duplicative or contradictory information in those interpretations. To explore these limitations, we performed a meta-analysis of optical surrogates for DOM from diverse sources, both from natural systems and after water treatment processes (n = 762). Prior to analysis, data were screened using a newly developed, standardized methodology that applies systematic quality control criteria before reporting surrogates. There was substantial overlap in surrogate values from natural and treated samples, suggesting that the gradients governing the surrogate variability can be generated in both contexts. This overlap provides justification for using optical surrogates originally developed in the context of natural systems to describe DOM changes in engineered systems, although the interpretations may change. Absorbance-based surrogates that describe the amount of spectral tailing (e.g., E2:E3 and S275-295) had a high frequency of strong correlations with one another but not to specific absorbance (SUVA254) or absorbance slope ratio (SR). The fluorescence index (FI) and biological index (ß/α) were strongly correlated with one another and to the peak emission wavelength but not to the humification index (HIX). Although SUVA254 and FI have both been correlated to DOM aromaticity in prior research, there was a lack of reciprocity between these optical surrogates across this data set. Additionally, there were patterns of deviations in the wastewater subset, suggesting that effluent organic matter may not follow conventional interpretations, urging caution in the use of optical surrogates to track DOM in water reuse applications. Finally, the meta-analysis highlights that three aspects should be captured when optical spectra are used for DOM interpretation: specific absorbance, absorbance tailing, and the extent of red-shifted fluorescence. We recommend that SUVA254, E2:E3, and FI or ß/α be prioritized in future DOM studies to capture these aspects, respectively.
Assuntos
Compostos Orgânicos , Compostos Orgânicos/química , Purificação da ÁguaRESUMO
Concordance occurs when two or more biological groups are correlated to each other. Examining the degree of concordance between communities has been a central goal in ecology. However, few studies have assessed the levels of community concordance at large spatial scales. We used a dataset obtained by the National Lakes Assessment (United States Environmental Protection Agency) to evaluate whether (i) the levels of concordance between aquatic communities were higher at the continental scale than within individual ecoregions of the United States and (ii) whether the levels of concordance between phytoplankton and zooplankton were higher than those between the plankton and macroinvertebrates communities. At the continental scale, the levels of concordance between different pairs of aquatic communities were low and did not exceed those within the ecoregions. Furthermore, levels of concordance varied considerably among ecoregions. Our results suggest that interactions between aquatic communities likely determined concordance patterns; however, the expectation of higher levels of concordance between the phytoplankton and zooplankton communities than between them and the macroinvertebrates community was not supported. The consistently low and variable levels of concordance suggest that using surrogate groups is not recommendable for monitoring lakes in the United States, both at the continental and regional scales. According to our results, the prospect of using the surrogacy approach was low even for aquatic communities that are highly interactive or driven by similar environmental factors.
Assuntos
Lagos , Fitoplâncton , Zooplâncton , Estados Unidos , Animais , Invertebrados , EcossistemaRESUMO
Conserving Nature's Stage (CNS) is a concept from conservation planning that promotes the protection of areas encompassing a broad range of enduring geophysical traits to provide long-term habitat for diverse species. The efficacy of using enduring geophysical characteristics as surrogates for biodiversity, independent of non-geophysical features and when considering finer resolution area selections, has yet to be investigated. Here, we evaluated CNS using 33 fine-scale inventories of vascular plant, non-vascular plant, invertebrate or vertebrate species from 13 areas across three continents. For each inventory, we estimated a continuous multidimensional surrogate defined from topographic and soil estimates of the surveyed plots. We assessed surrogate effectiveness by comparing the species representation of surrogate selected plots to the representation from plots picked randomly and using species information. We then used correlation coefficients to assess the link between the performance and qualities of the inventories, surroundings and surrogates. The CNS surrogate showed positive performance for 24 of the 33 inventories, and among these tests, represented 28 more species than random and 83% of the total number of species on average. We also found a small number of weak correlations between performance and environmental variability, as well as qualities of the surrogate. Our study demonstrates that prioritizing areas for a variety of geophysical characteristics will, in most cases, promote the representation of species. Our findings also point to areas for future research that might enhance CNS surrogacy. This article is part of the Theo Murphy meeting issue 'Geodiversity for science and society'.
Assuntos
Biodiversidade , Conservação dos Recursos Naturais , Ecossistema , SoloRESUMO
AIMS: This study aimed to compare the heat inactivation kinetics of viable human norovirus with the surrogate, MS2 bacteriophage as well as assess the decay of the RNA signal. METHODS AND RESULTS: Human intestinal enteroids were used to analyze the heat inactivation kinetics of viable human norovirus compared to the surrogate MS2 bacteriophage, which was cultured using a plaque assay. Norovirus decay rates were 0.22 min-1, 0.68 min-1, and 1.11 min-1 for 50°C, 60°C, and 70°C, respectively, and MS2 bacteriophage decay rates were 0.0065 min-1, 0.045 min-1, and 0.16 min-1 for 50°C, 60°C, and 70°C, respectively. Norovirus had significantly higher decay rates than MS2 bacteriophage at all tested temperatures (P = .002-.007). No decrease of RNA titers as measured by reverse transcription-PCR for both human norovirus and MS2 bacteriophage over time was observed, indicating molecular methods do not accurately depict viable human norovirus after heat inactivation and treatment efficiency is underestimated. CONCLUSIONS: Overall, our data demonstrate that MS2 bacteriophage is a conservative surrogate to measure heat inactivation and potentially overestimates the infectious risk of norovirus. Furthermore, this study corroborates that measuring viral RNA titers, as evaluated by PCR methods, does not correlate with the persistence of viable norovirus under heat inactivation.
Assuntos
Norovirus , Humanos , Norovirus/genética , Temperatura Alta , Levivirus/genética , RNA Viral/genética , Cinética , Inativação de VírusRESUMO
The risk of the use of toxic chemicals for unlawful acts has been a matter of concern for different governments and multilateral agencies. The Organisation for the Prohibition of Chemical Weapons (OPCW), which oversees the implementation of the Chemical Weapons Convention (CWC), considering recent events employing chemical warfare agents as means of assassination, has recently included in the CWC "Annex on Chemicals" some organophosphorus compounds that are regarded as acting in a similar fashion to the classical G- and V-series of nerve agents, inhibiting the pivotal enzyme acetylcholinesterase. Therefore, knowledge of the activity of the pyridinium oximes, the sole class of clinically available acetylcholinesterase reactivators to date, is plainly justified. In this paper, continuing our research efforts in medicinal chemistry on this class of toxic chemicals, we synthesized an A-230 nerve agent surrogate and applied a modified Ellman's assay to evaluate its ability to inhibit our enzymatic model, acetylcholinesterase from Electrophorus eel, and if the clinically available antidotes are able to rescue the enzyme activity for the purpose of relating the findings to the previously disclosed in silico data for the authentic nerve agent and other studies with similar A-series surrogates. Our experimental data indicates that pralidoxime is the most efficient compound for reactivating acetylcholinesterase inhibited by A-230 surrogate, which is the opposite of the in silico data previously disclosed.
RESUMO
Plasma is an important biofluid for clinical research and diagnostics. In the clinic, unpredictable delays-from minutes to hours-between blood collection and plasma generation are often unavoidable. These delays can potentially lead to protein degradation and modification and might considerably affect intact protein measurement methods such as sandwich enzyme-linked immunosorbent assays that bind proteins on two epitopes to increase specificity, thus requiring largely intact protein structures. Here, we investigated, using multiple reaction monitoring mass spectrometry (MRM-MS), how delays in plasma processing affect peptide-centric "bottom-up" proteomics. We used validated assays for proteotypic peptide surrogates of 270 human proteins to analyze plasma generated after whole blood had been kept at room temperature from 0 to 40 h to mimic delays that occur in the clinic. Moreover, we evaluated the impact of different plasma-thawing conditions on MRM-based plasma protein quantitation. We demonstrate that >90% of protein concentration measurements were unaffected by the thawing procedure and by up to 40-h delayed plasma generation, reflected by relative standard deviations (RSDs) of <30%. Of the 159 MRM assays that yielded quantitative results in 60% of the measured time points, 139 enabled a stable protein quantitation (RSD <20%), 14 showed a slight variation (RSD 20-30%), and 6 appeared unstable/irreproducible (RSD > 30%). These results demonstrate the high robustness and thus the potential for MRM-based plasma-protein quantitation to be used in a clinical setting. In contrast to enzyme-linked immunosorbent assay, peptide-based MRM assays do not require intact three-dimensional protein structures for an accurate and precise quantitation of protein concentrations in the original sample.
Assuntos
Proteínas Sanguíneas , Proteômica , Proteínas Sanguíneas/análise , Ensaio de Imunoadsorção Enzimática , Humanos , Espectrometria de Massas/métodos , Peptídeos/análise , Proteômica/métodosRESUMO
Protein quantitation via mass spectrometry relies on peptide proxies for the parent protein from which abundances are estimated. Owing to the variability in signal from individual peptides, accurate absolute quantitation usually relies on the addition of an external standard. Typically, this involves stable isotope-labeled peptides, delivered singly or as a concatenated recombinant protein. Consequently, the selection of the most appropriate surrogate peptides and the attendant design in recombinant proteins termed QconCATs are challenges for proteome science. QconCATs can now be built in a "a-la-carte" assembly method using synthetic biology: ALACATs. To assist their design, we present "AlacatDesigner", a tool that supports the peptide selection for recombinant protein standards based on the user's target protein. The user-customizable tool considers existing databases, occurrence in the literature, potential post-translational modifications, predicted miscleavage, predicted divergence of the peptide and protein quantifications, and ionization potential within the mass spectrometer. We show that peptide selections are enriched for good proteotypic and quantotypic candidates compared to empirical data. The software is freely available to use either via a web interface AlacatDesigner, downloaded as a Desktop application or imported as a Python package for the command line interface or in scripts.
Assuntos
Peptídeos , Software , Peptídeos/química , Espectrometria de Massas , Proteoma/metabolismo , Proteínas RecombinantesRESUMO
Development of heterogeneous catalysts with tunable activity and selectivity has posed a persistent challenge. This study addresses this challenge by fabricating a hybrid environment through the combination of mesoporous silica and N-rich melamine dendron via covalent grafting, allowing for controllable growth and encapsulation of Pd NPs. This catalyst presented an excellent catalytic activity for the oxidative carbonylative self-coupling of aryl boronic acids to afford symmetric biaryl ketones using N-formyl saccharin as a sustainable solid CO source and Cu as a co-catalyst.
RESUMO
PURPOSE OF REVIEW: Non-invasive measurements such as arterial stiffness serve as proxy surrogates for detection of early atherosclerosis and ASCVD risk stratification. These surrogate measurements are influenced by age, gender, and ethnicity and affected by the physiological changes of puberty and somatic growth in children and adolescents. RECENT FINDINGS: There is no consensus of the ideal method to measure surrogate markers in youth (< 18 years of age), nor standardized imaging protocols for youth. Currently, pediatric normative data are available but not generalizable. In this review, we provide rationale on how currently used surrogates can help identify subclinical atherosclerosis in youth and affirm their role in identifying youth at risk for premature CVD.
Assuntos
Aterosclerose , Doenças Cardiovasculares , Rigidez Vascular , Humanos , Adolescente , Criança , Doenças Cardiovasculares/diagnóstico , Artérias Carótidas , Fatores de Risco , Aterosclerose/diagnóstico , Biomarcadores , Espessura Intima-Media CarotídeaRESUMO
BACKGROUND: Obesity and metabolic syndrome are observed more frequently in infertile women, and insulin resistance (IR) is closely related to them. However, there are no studies that have examined the association between different IR surrogates and female infertility, hence we investigated the potential association between them in the general population. METHODS: This was a cross-sectional study using data from the National Health and Nutrition Examination Survey (NHANES, 2013-2018). The association of different IR surrogates (HOMA-IR index, TyG index and TyG-BMI index) with female infertility was estimated by multivariable regression analysis. RESULTS: After adjusting for confounders, the HOMA-IR index and TyG index did not show an association with female infertility, while the TyG-BMI index was found to have a positive association with female infertility (OR = 1.01, 95% CI: 1.00, 1.01; P < 0.0001), and the OR of the TyG-BMI group T3 (≥ 255.55) was significantly different compared to the group T1 (< 185.31) (OR = 3.02, 95% CI: 1.62, 5.60). Similar results were seen in most of the subgroup participants by stratified analysis (P-interaction > 0.05). However, different IR surrogates did not show variability in their ability to predict infertility [TyG-BMI: 0.68 (95% CI: 0.62, 0.74) vs. TyG: 0.62 (95% CI: 0.57, 0.68) vs. HOMA-IR: 0.65 (95% CI: 0.60, 0.71)]. CONCLUSIONS: Our result suggests that high levels of TyG-BMI index were positively associated with female infertility in US reproductive-aged females.
Assuntos
Infertilidade Feminina , Resistência à Insulina , Humanos , Feminino , Adulto , Infertilidade Feminina/epidemiologia , Inquéritos Nutricionais , Glicemia/análise , Estudos Transversais , Biomarcadores , Triglicerídeos , GlucoseRESUMO
PURPOSE: To examine surrogates' mental health, social support, and relationship with intended parents (IPs) during the COVID-19 pandemic from March 2020 to February 2022. METHODS: Data were collected between April 29, 2022 and July 31, 2022, at an academic IVF center in Canada using an 85-item online anonymous cross-sectional survey that included three standardized scales measuring mental health (PHQ-4), loneliness, and social support. Eligible surrogates actively involved in surrogacy during the study period received email invitations. RESULTS: The response rate was 50.3% (338/672); 320 submitted surveys were analyzed. Two-thirds (65%) of respondents experienced mental health concerns during the pandemic and were significantly less comfortable about seeking mental health support than those without concerns. Nonetheless, 64% were highly satisfied with their surrogacy experience; 80% received a high level of support from their IPs, and 90% reported a good relationship with them. The final hierarchical regression model identified five significant predictors, explaining 39.4% of the variance in PHQ-4 scores: a prior mental health history, COVID-19 impact on personal life, surrogacy satisfaction, loneliness, and social support. CONCLUSIONS: COVID-19 created an unprecedented challenge to surrogacy care, increasing surrogates' risk of experiencing mental health symptoms. Our data show that IP support and the surrogate-IP relationship were fundamentals to surrogacy satisfaction. The findings are relevant to fertility and mental health practitioners in identifying surrogates who are more susceptible to mental health challenges. Fertility clinics should ensure adequate psychological screening of surrogate candidates and proactively offer mental health support services.
Assuntos
COVID-19 , Pandemias , Gravidez , Feminino , Humanos , Saúde Mental , Estudos Transversais , Mães Substitutas/psicologia , Relações Interpessoais , COVID-19/epidemiologia , Apoio SocialRESUMO
The detection of infectious SARS-CoV-2 in food and food packaging associated with the cold chain has raised concerns about the possible transmission pathway of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in foods transported through cold-chain logistics and the need for novel decontamination strategies. In this study, the effect of electron beam (E-beam) irradiation on the inactivation of two SARS-CoV-2surrogate, viruses porcine epidemic diarrhea virus (PEDV) and porcine transmissible gastroenteritis virus (TGEV), in culture medium and food substrate, and on food substrate were investigated. The causes of virus inactivation were also investigated by transmission electron microscopy (TEM) and Quantitative Real-time PCR (QRT-PCR). Samples packed inside and outside, including virus-inoculated large yellow croaker and virus suspensions, were irradiated with E-beam irradiation (2, 4, 6, 8, 10 kGy) under refrigerated (0 °C)and frozen (-18 °C) conditions. The titers of both viruses in suspension and fish decreased significantly (P < 0.05) with increasing doses of E-beam irradiation. The maximum D10 value of both viruses in suspension and fish was 1.24 kGy. E-beam irradiation at doses below 10 kGy was found to destroy the spike proteins of both SARS-CoV-2 surrogate viruses by transmission electron microscopy (TEM) and negative staining of thin-sectioned specimens, rendering them uninfectious. E-beam irradiation at doses greater than 10 kGy was also found to degrade viral genomic RNA by qRT-PCR. There were no significant differences in color, pH, TVB-N, TBARS, and sensory properties of irradiated fish samples at doses below 10 kGy. These findings suggested that E-beam irradiation has the potential to be developed as an efficient non-thermal treatment to reduce SARS-CoV-2 contamination in foods transported through cold chain foods to reduce the risk of SARS-CoV-2 infection in humans through the cold chain.
RESUMO
The study of peptides (synthetic or corresponding to discrete regions of proteins) has facilitated the understanding of protein structure-activity relationships. Short peptides can also be used as powerful therapeutic agents. However, the functional activity of many short peptides is usually substantially lower than that of their parental proteins. This is (as a rule) due to their diminished structural organization, stability, and solubility often leading to an enhanced propensity for aggregation. Several approaches have emerged to overcome these limitations, which are aimed at imposing structural constraints into the backbone and/or sidechains of the therapeutic peptides (such as molecular stapling, peptide backbone circularization and molecular grafting), therefore enforcing their biologically active conformation and thus improving their solubility, stability, and functional activity. This review provides a short summary of approaches aimed at enhancing the biological activity of short functional peptides with a particular focus on the peptide grafting approach, whereby a functional peptide is inserted into a scaffold molecule. Intra-backbone insertions of short therapeutic peptides into scaffold proteins have been shown to enhance their activity and render them a more stable and biologically active conformation.
Assuntos
Peptídeos , Peptídeos/química , Conformação Molecular , Conformação ProteicaRESUMO
Chemical synthesis of insulin superfamily proteins (ISPs) has recently been widely studied to develop next-generation drugs. Separate synthesis of multiple peptide fragments and tedious chain-to-chain folding are usually encountered in these studies, limiting accessibility to ISP derivatives. Here we report the finding that insulin superfamily proteins (e.g. H2 relaxin, insulin itself, and H3 relaxin) incorporating a pre-made diaminodiacid bridge at A-B chain terminal disulfide can be easily and rapidly synthesized by a single-shot automated solid-phase synthesis and expedient one-step folding. Our new H2 relaxin analogues exhibit almost identical structures and activities when compared to their natural counterparts. This new synthetic strategy will expediate production of new ISP analogues for pharmaceutical studies.
Assuntos
Relaxina , Relaxina/química , Relaxina/metabolismo , Dissulfetos/química , Técnicas de Síntese em Fase Sólida , Proteínas/química , Insulina/química , Receptores Acoplados a Proteínas G/metabolismoRESUMO
Population-scale expression profiling studies can provide valuable insights into biological and disease-underlying mechanisms. The availability of phenotypic traits is essential for studying clinical effects. Therefore, missing, incomplete, or inaccurate phenotypic information can make analyses challenging and prevent RNA-seq or other omics data to be reused. A possible solution are predictors that infer clinical or behavioral phenotypic traits from molecular data. While such predictors have been developed based on different omics data types and are being applied in various studies, metabolomics-based surrogates are less commonly used than predictors based on DNA methylation profiles.In this study, we inferred 17 traits, including diabetes status and exposure to lipid medication, using previously trained metabolomic predictors. We evaluated whether these metabolomic surrogates can be used as an alternative to reported information for studying the respective phenotypes using expression profiling data of four population cohorts. For the majority of the 17 traits, the metabolomic surrogates performed similarly to the reported phenotypes in terms of effect sizes, number of significant associations, replication rates, and significantly enriched pathways.The application of metabolomics-derived surrogate outcomes opens new possibilities for reuse of multi-omics data sets. In studies where availability of clinical metadata is limited, missing or incomplete information can be complemented by these surrogates, thereby increasing the size of available data sets. Additionally, the availability of such surrogates could be used to correct for potential biological confounding. In the future, it would be interesting to further investigate the use of molecular predictors across different omics types and cohorts.
Assuntos
Metabolômica , FenótipoRESUMO
Multiple pathways of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission have been examined, and the role of contaminated foods as a source of SARS-CoV-2 exposure has been suggested. As many cases of SARS-CoV-2 have been linked to meat processing plants, it may be that conditions in live animal markets and slaughterhouses or meat processing plant procedures transfer viral particles to meat, poultry, and seafood during animal slaughter, processing, storage, or transport. Because of the potential for contamination of foods such as beef, chicken, pork, or fish, the goal of this study was to evaluate the survival of a lipid enveloped RNA bacteriophage, phi 6, as well as two animal coronaviruses, murine hepatitis virus (MHV) and transmissible gastroenteritis virus (TGEV), as SARS-CoV-2 surrogates for their survival under various meat and fish cold-storage conditions over 30 days. Viral surrogates differed in survival, depending on food product and temperature, but overall, viruses survived for extended periods of time at high concentrations at both refrigerated and frozen temperatures. The ability of SARS-CoV-2 viral surrogates like Phi 6 and animal coronaviruses to survive for varying extents on some meat and fish products when stored refrigerated or frozen is a significant and concerning finding. Continued efforts are needed to prevent contamination of foods and food processing surfaces, worker hands, and food processing utensils such as knives, and there is a need to better address the lack of or inadequate disinfection of these foods prior to meat packaging. IMPORTANCE The ability of SARS-CoV-2 viral surrogates like Phi 6 and animal coronaviruses to survive for long periods on meat and fish products at cold temperatures emphasizes the need for rigorous and sustained food sanitation and hygiene in the harvest, transport, processing, and distribution of these foods.
Assuntos
COVID-19 , Vírus da Hepatite Murina , Animais , Bovinos , Produtos Pesqueiros , Carne , Camundongos , SARS-CoV-2RESUMO
OBJECTIVE: Digital microscopy is used to monitor particulates such as protein aggregates within biopharmaceutical products. The images that result encode a wealth of information that is underutilized in pharmaceutical process monitoring. For example, images of particles in protein drug products typically are analyzed only to obtain particle counts and size distributions, even though the images also reflect particle characteristics such as shape and refractive index. Multiple groups have demonstrated that convolutional neural networks (CNNs) can extract information from images of protein aggregates allowing assignment of the likely stress at the "root-cause" of aggregation. A practical limitation of previous CNN-based approaches is that the potential aggregation-inducing stresses must be known a priori, disallowing identification of particles produced by unknown stresses. METHODS: We demonstrate an expanded CNN analysis of flow imaging microscopy (FIM) images incorporating judiciously chosen particle standards within a recently proposed "fingerprinting algorithm" (Biotechnol. & Bioeng. (2020) 117:3322) that allows detection of particles formed by unknown root-causes. We focus on ethylene tetrafluoroethylene (ETFE) microparticles as standard surrogates for protein aggregates. We quantify the sensitivity of the new algorithm to experimental parameters such as microscope focus and solution refractive index changes, and explore how FIM sample noise affects statistical testing procedures. RESULTS & CONCLUSIONS: Applied to real-world microscopy images of protein aggregates, the algorithm reproducibly detects complex, distinguishing "textural features" of particles that are not easily described by standard morphological measurements. This offers promise for quality control applications and for detecting shifts in protein aggregate populations due to stresses resulting from unknown process upsets.