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The Last Common Ancestor (LCA) is understood as a hypothetical population of organisms from which all extant living creatures are thought to have descended. Its biology and environment have been and continue to be the subject of discussions within the scientific community. Since the first bacterial genomes were obtained, multiple attempts to reconstruct the genetic content of the LCA have been made. In this review, we compare 10 of the most extensive reconstructions of the gene content possessed by the LCA as they relate to aspects of the translation machinery. Although each reconstruction has its own methodological biases and many disagree in the metabolic nature of the LCA all, to some extent, indicate that several components of the translation machinery are among the most conserved genetic elements. The datasets from each reconstruction clearly show that the LCA already had a largely complete translational system with a genetic code already in place and therefore was not a progenote. Among these features several ribosomal proteins, transcription factors like IF2, EF-G, and EF-Tu and both class I and class II aminoacyl tRNA synthetases were found in essentially all reconstructions. Due to the limitations of the various methodologies, some features such as the occurrence of rRNA posttranscriptional modified bases are not fully addressed. However, conserved as it is, non-universal ribosomal features found in various reconstructions indicate that LCA's translation machinery was still evolving, thereby acquiring the domain specific features in the process. Although progenotes from the pre-LCA likely no longer exist recent results obtained by unraveling the early history of the ribosome and other genetic processes can provide insight to the nature of the pre-LCA world.
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Bacterias , Evolución Molecular , Biosíntesis de Proteínas , Biosíntesis de Proteínas/genética , Bacterias/genética , Bacterias/metabolismo , Filogenia , Código Genético , Aminoacil-ARNt Sintetasas/genética , Aminoacil-ARNt Sintetasas/metabolismo , Proteínas Ribosómicas/genética , Ribosomas/genética , Ribosomas/metabolismo , Genoma BacterianoRESUMEN
One of the central issues in the understanding of early cellular evolution is the characterisation of the cenancestor. This includes the description of the chemical nature of its genome. The disagreements on this question comprise several proposals, including the possibility that AlkB-mediated methylation repair of alkylated RNA molecules may be interpreted as evidence of a cenancestral RNA genome. We present here an evolutionary analysis of the cupin-like protein superfamily based on tertiary structure-based phylogenies that includes the oxygen-dependent AlkB and its homologs. Our results suggest that the repair of methylated RNA molecules is the outcome of the enzyme substrate ambiguity, and doesn´t necessarily indicates that the last common ancestor was endowed with an RNA genome.
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ADN , Evolución Molecular , Genoma , Filogenia , ARN , ARN/genética , Genoma/genética , ADN/genética , Enzimas AlkB/genética , Enzimas AlkB/metabolismo , MetilaciónRESUMEN
BACKGROUND: Bile acids (BAs) are steroid-derived molecules with important roles in digestion, the maintenance of host metabolism, and immunomodulation. Primary BAs are synthesized by the host, while secondary BAs are produced by the gut microbiome through transformation of the former. The regulation of microbial production of secondary BAs is not well understood, particularly the production of 7-dehydroxylated BAs, which are the most potent agonists for host BA receptors. The 7-dehydroxylation of cholic acid (CA) is well established and is linked to the expression of a bile acid-inducible (bai) operon responsible for this process. However, little to no 7-dehydroxylation has been reported for other host-derived BAs (e.g., chenodeoxycholic acid, CDCA or ursodeoxycholic acid, UDCA). RESULTS: Here, we demonstrate that the 7-dehydroxylation of CDCA and UDCA by the human isolate Clostridium scindens is induced when CA is present, suggesting that CA-dependent transcriptional regulation is required for substantial 7-dehydroxylation of these primary BAs. This is supported by the finding that UDCA alone does not promote expression of bai genes. CDCA upregulates expression of the bai genes but the expression is greater when CA is present. In contrast, the murine isolate Extibacter muris exhibits a distinct response; CA did not induce significant 7-dehydroxylation of primary BAs, whereas BA 7-dehydroxylation was promoted upon addition of germ-free mouse cecal content in vitro. However, E. muris was found to 7-dehydroxylate in vivo. CONCLUSIONS: The distinct expression responses amongst strains indicate that bai genes are regulated differently. CA promoted bai operon gene expression and the 7-dehydroxylating activity in C. scindens strains. Conversely, the in vitro activity of E. muris was promoted only after the addition of cecal content and the isolate did not alter bai gene expression in response to CA. The accessory gene baiJ was only upregulated in the C. scindens ATCC 35704 strain, implying mechanistic differences amongst isolates. Interestingly, the human-derived C. scindens strains were also capable of 7-dehydroxylating murine bile acids (muricholic acids) to a limited extent. This study shows novel 7-dehydroxylation activity in vitro resulting from the presence of CA and suggests distinct bai gene expression across bacterial species.
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Ácidos y Sales Biliares , Ácido Cólico , Ácido Cólico/metabolismo , Animales , Ácidos y Sales Biliares/metabolismo , Ratones , Humanos , Clostridium/metabolismo , Clostridium/genética , Regulación Bacteriana de la Expresión Génica , Hidroxilación , Operón , Ácido Quenodesoxicólico/metabolismo , Ácido Ursodesoxicólico/metabolismo , Microbioma GastrointestinalRESUMEN
BACKGROUND & OBJECTIVE: The progression of dengue fever to severe dengue (SD) is a major public health concern that impairs the capacity of the medical system to predict and treat dengue patients. Hence, the present study used a metabolomic approach integrated with machine models to identify differentially expressed metabolites in patients with SD compared to nonsevere patients and healthy controls. METHODS: Comprehensively, the plasma was collected at different clinical phases during dengue without warning signs (DWOW, N = 10), dengue with warning signs (DWW, N = 10), and SD (N = 10) at different stages [i.e., day of admission (DOA), day of defervescence (DOD), and day of convalescent (DOC)] in comparison to healthy control (HC). The samples were subjected to LCâESIâMS/MS to identify metabolites. Statistical and machine learning analyses were performed using R and Python language. Further, biomarker, pathway and correlation analysis was performed to identify potential predictors of dengue. RESULTS & CONCLUSION: A total of 423 metabolites were identified in all the study groups. Paired and unpaired t-tests revealed 14 highly differentially expressed metabolites between and across the dengue groups, with four metabolites (shikimic acid, ureidosuccinic acid, propionyl carnitine, and alpha-tocopherol) showing significant differences compared to HC. Furthermore, biomarker (ROC) analysis revealed 11 potential molecules with a significant AUC value of 1 that could serve as potential biomarkers for identifying different dengue clinical stages that are beneficial for predicting dengue disease outcomes. The logistic regression model revealed that S-adenosylhomocysteine, hypotaurine, and shikimic acid metabolites could be beneficial indicators for predicting severe dengue, with an accuracy and AUC of 0.75. The data showed that dengue infection is related to lipid metabolism, oxidative stress, inflammation, metabolomic adaptation, and virus manipulation. Moreover, the biomarkers had a significant correlation with biochemical parameters like platelet count, and hematocrit. These results shed some light on host-derived small-molecule biomarkers that are associated with dengue severity and novel insights into metabolomics mechanisms interlinked with disease severity.
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Biomarcadores , Dengue , Aprendizaje Automático , Metabolómica , Espectrometría de Masas en Tándem , Humanos , Metabolómica/métodos , Dengue/metabolismo , Dengue/sangre , Espectrometría de Masas en Tándem/métodos , Masculino , Biomarcadores/sangre , Biomarcadores/metabolismo , Femenino , Cromatografía Liquida/métodos , Adulto , Metaboloma , Persona de Mediana Edad , Cromatografía Líquida con Espectrometría de MasasRESUMEN
Rapidly increasing heavy metal waste has adversely affected the environment and the Earth's health. The lack of appropriate remediation technologies has worsened the issue globally, especially in developing countries. Heavy-metals contaminants have severely impacted the environment and led to devastating conditions owing to their abundance and reactivity. As they are nondegradable, the potential risk increases even at a low concentration. However, heavy-metal remediation has increased with the up-gradation of technologies and integration of new approaches. Also, of all the treatment methodologies, microbial-assisted multifaceted approach for ameliorating heavy metals is a promising strategy for propagating the idea of a green and sustainable environment with minimal waste aggregation. Microbial remediation combined with different biotechniques could aid in unraveling new methods for eradicating heavy metals. Thus, the present review focuses on various microbial remediation approaches and their affecting factors, enabling recapitulation of the interplay between heavy-metals ions and microorganisms. Additionally, heavy-metals remediation mechanisms adapted by microorganisms, the role of genetically modified (GM) microorganisms, life cycle assessment (LCA), techno-economic assessment (TEA) limitations, and prospects of microbial-assisted amelioration of heavy-metals have been elaborated in the current review with focus toward "sustainable and greener future."
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Metales Pesados , Contaminantes del Suelo , Biodegradación Ambiental , Contaminantes del Suelo/análisisRESUMEN
Diabetic macular edema (DME) is the most common cause of blindness in patients with diabetic retinopathy. To investigate the proteomic profiles of the aqueous humor (AH) of individuals with diabetic macular edema (DME), AH samples were collected from patients with non-diabetes mellitus (NDM), DM, nonproliferative diabetic retinopathy (NPDR), and DME. We performed comparative proteomic analyses using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and bioinformatics analyses. We identified 425 proteins in these AH samples, of which 113 showed changes in expression in DME compared with NDM, 95 showed changes in expression in DME vs. DM, and 84 showed changes in expression in DME compared with NPDR. The bioinformatics analysis suggested that DME is closely associated with platelet degranulation, oxidative stress-related pathway, and vascular-related pathways. Upregulation of haptoglobin (HP) and downregulation of fibrillin 1 (FBN1) were validated by ELISA. Receiver operating characteristic (ROC) analysis showed that HP and FBN1 could distinguish DME from NPDR with areas under the curve of 0.987 (p = 0.00608) and 0.791 (p = 0.00629), respectively. The findings provide potential clues for further analysis of the molecular mechanisms and the development of new treatments for DME. HP and FBN1 may be potential key proteins and therapeutic targets in human DME. The proteomics dataset generated has been deposited to the ProteomeXchange/iProX Consortium with Identifier: PXD033404/IPX0004353001.
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Diabetes Mellitus , Retinopatía Diabética , Edema Macular , Humanos , Retinopatía Diabética/metabolismo , Edema Macular/metabolismo , Humor Acuoso/metabolismo , Proteómica/métodos , Cromatografía Liquida , Espectrometría de Masas en Tándem , Diabetes Mellitus/metabolismoRESUMEN
Nitro fatty acids (NO2-FAs) are biologically active compounds produced from the reaction of unsaturated fatty acids with reactive nitrogen species (RNS). Due to their electrophilic nature, these endogenously produced metabolites can react with nucleophilic targets, producing a spectrum of modulatory and protective effects. Determination of NO2-FAs in biological samples is challenging due to their low nanomolar to picomolar endogenous concentrations, indistinct metabolism, and distribution in many tissues and biofluids. Several attempts have been made to develop precise, standardized, and efficient methodologies for assessing physiological and pathophysiological processes to overcome the difficulties associated with their measurement. This review discusses those approaches utilizing liquid chromatography tandem mass spectrometry (LCâMS/MS) and gas chromatography tandem mass spectrometry (GCâMS/MS) for the quantification of NO2-FAs, in addition to a summary of their laboratory synthesis and extraction from biological samples. Clinical associations with different pathological conditions, including hyperlipidaemia, cardiac ischemia and herpes simplex type 2 viral infection (HSV-2), are also discussed.
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Ácidos Grasos , Espectrometría de Masas en Tándem , Humanos , Ácidos Grasos/análisis , Espectrometría de Masas en Tándem/métodos , Cromatografía Liquida/métodos , Cromatografía de Gases y Espectrometría de Masas/métodos , Animales , Nitrocompuestos/análisis , Nitrocompuestos/químicaRESUMEN
Trichinella spiralis (T. spiralis) is a zoonotic parasitic nematode with a unique life cycle, as all developmental stages are contained within a single host. Excretory-secretory (ES) proteins are the main targets of the interactions between T. spiralis and the host at different stages of development and are essential for parasite survival. However, the ES protein profiles of T. spiralis at different developmental stages have not been characterized. The proteomes of ES proteins from different developmental stages, namely, muscle larvae (ML), intestinal infective larvae (IIL), preadult (PA) 6 h, PA 30 h, adult (Ad) 3 days post-infection (dpi) and Ad 6 dpi, were characterized via label-free mass spectrometry analysis in combination with bioinformatics. A total of 1217 proteins were identified from 9341 unique peptides in all developmental stages, 590 of which were quantified and differentially expressed. GO classification and KEGG pathway analysis revealed that these proteins were important for the growth of the larvae and involved in energy metabolism. Moreover, the heat shock cognate 71 kDa protein was the centre of protein interactions at different developmental stages. The results of this study provide comprehensive proteomic data on ES proteins and reveal that these ES proteins were differentially expressed at different developmental stages. Differential proteins are associated with parasite survival and the host immune response and may be potential early diagnostic antigen or antiparasitic vaccine candidates.
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Trichinella spiralis , Trichinella , Triquinelosis , Animales , Triquinelosis/veterinaria , Proteínas del Helminto/metabolismo , Proteómica , Músculos , Larva/metabolismo , Antígenos Helmínticos , Trichinella/metabolismoRESUMEN
OBJECTIVES: Public Health officials are often challenged to effectively allocate limited resources. Social determinants of health (SDOH) may cluster in areas to cause unique profiles related to various adverse life events. The authors use the framework of unintended teen pregnancies to illustrate how to identify the most vulnerable neighborhoods. METHODS: This study used data from the U.S. American Community Survey, Princeton Eviction Lab, and Connecticut Office of Vital Records. Census tracts are small statistical subdivisions of a county. Latent class analysis (LCA) was employed to separate the 832 Connecticut census tracts into four distinct latent classes based on SDOH, and GIS mapping was utilized to visualize the distribution of the most vulnerable neighborhoods. GEE Poisson regression model was used to assess whether latent classes were related to the outcome. Data were analyzed in May 2021. RESULTS: LCA's results showed that class 1 (non-minority non-disadvantaged tracts) had the least diversity and lowest poverty of the four classes. Compared to class 1, class 2 (minority non-disadvantaged tracts) had more households with no health insurance and with single parents; and class 3 (non-minority disadvantaged tracts) had more households with no vehicle available, that had moved from another place in the past year, were low income, and living in renter-occupied housing. Class 4 (minority disadvantaged tracts) had the lowest socioeconomic characteristics. CONCLUSIONS: LCA can identify unique profiles for neighborhoods vulnerable to adverse events, setting up the potential for differential intervention strategies for communities with varying risk profiles. Our approach may be generalizable to other areas or other programs. KEY MESSAGES: What is already known on this topic Public health practitioners struggle to develop interventions that are universally effective. The teen birth rates vary tremendously by race and ethnicity. Unplanned teen pregnancy rates are related to multiple social determinants and behaviors. Latent class analysis has been applied successfully to address public health problems. What this study adds While it is the pregnancy that is not planned rather than the birth, access to pregnancy intention data is not available resulting in a dependency on teen birth data for developing public health strategies. Using teen birth rates to identify at-risk neighborhoods will not directly represent the teens at risk for pregnancy but rather those who delivered a live birth. Since teen birth rates often fluctuate due to small numbers, especially for small neighborhoods, LCA may avoid some of the limitations associated with direct rate comparisons. The authors illustrate how practitioners can use publicly available SDOH from the Census Bureau to identify distinct SDOH profiles for teen births at the census tract level. How this study might affect research, practice or policy These profiles of classes that are at heightened risk potentially can be used to tailor intervention plans for reducing unintended teen pregnancy. The approach may be adapted to other programs and other states to prioritize the allocation of limited resources.
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Sistemas de Información Geográfica , Análisis de Clases Latentes , Determinantes Sociales de la Salud , Humanos , Femenino , Adolescente , Embarazo , Connecticut , Características del Vecindario , Poblaciones Vulnerables/estadística & datos numéricos , Características de la Residencia/estadística & datos numéricos , Embarazo en Adolescencia/estadística & datos numéricos , Estados Unidos , Factores SocioeconómicosRESUMEN
The ability to provide adequate nutrition is considered a key factor in evaluating the sustainability of foods and diets. Nutrient indices are used as functional units (FU) in life cycle assessment of foods to include nutritional performance in the environmental assessment of a product. Several general and food-group-specific nutrient indices exist but many lack validation, particularly when used as FU. In addition, the nutrient selection strategies and reference units for nutrient intake can vary considerably among studies. To validate intake-based product-group-specific nutrient indices previously developed for protein (NR-FIprot) and carbohydrate (NR-FIcarb) foods and for fruits and vegetables (NR-FIveg), we applied principal component analysis to investigate correlations between nutrients in foods and dishes representing a typical Finnish diet. The reference amounts for meal components were based on a plate model that reflected Finnish dietary recommendations. The portion sizes for the different food groups were anchored at 100 g, 135 g and 350 g for proteins, carbohydrates and fruits/vegetables, respectively. Statistical modelling largely validated the NR-FI indices, highlighting protein foods as sources of niacin, vitamin B12 and Se, carbohydrate foods as sources of Mg, Fe and phosphorous, and fruits/vegetables as sources of potassium, vitamin K, vitamin C, fibre and thiamine. However, in contrast to the intake-based approach applied in NR-FIprot, the dietary recommendation-based validation process suggested that fruits and vegetables should be favoured as sources of riboflavin and vitamin B6.
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Dieta , Frutas , Nutrientes , Valor Nutritivo , Verduras , Finlandia , Humanos , Nutrientes/análisis , Frutas/química , Carbohidratos de la Dieta/análisis , Proteínas en la Dieta/análisis , Análisis de Componente Principal , AlimentosRESUMEN
Little is known whether engagement in sexual behaviors associated with potential HIV risks differs by subgroups of men who have sex with men (MSM), who are distinct regarding patterns of use of online tools for partner-seeking. Using latent class analysis, we revealed four classes of app-using MSM (n = 181, 18-34 y.o., 82.4% identified as White and non-Hispanic) residing in Central Kentucky: the Grindr/Tinder class; the Poly App Use class of MSM-oriented apps; the General Social Media class, and the Bumble class. Unadjusted penalized logistic regressions showed associations of the Poly App Use class with increased numbers of receptive anal sex partners and reporting condomless receptive anal sex. Adjusting for other covariates, poly app users versus others were more likely to be older (25-34 vs. 18-24, AOR = 3.81, 95%CI = 1.70-9.03), to report past six-month illicit drug use (AOR = 2.93, 95%CI = 1.25-7.43) and to have ever used pre-exposure prophylaxis (AOR = 2.79, 95%CI = 1.10-7.12). Poly app users had behavior profiles associated with an elevated HIV risk and also reported HIV-related protective behaviors likely indicating increased risk awareness among this class. Our findings warrant differentiation of behavior profiles by patterns of app use and suggest not to generalize sexual behaviors associated with potential HIV risks to all app-using MSM.
RESUMEN: Poco se sabe si la participación en comportamientos sexuales asociados con riesgos potenciales de VIH difiere según distintos subgrupos de hombres que tienen sexo con hombres (HSH) en base a patrones en el uso de herramientas online para la búsqueda de pareja. Mediante el análisis de clases latentes, identificamos cuatro clases de HSH que usan aplicaciones (n = 181, 1834 años, 82.4% identificados como Blancos y no-Hispanos) y residen en Kentucky Central: la clase Grindr/Tinder, la clase Poly App Use de aplicaciones orientadas a HSH, la clase General Social Media y la clase Bumble. Las regresiones logísticas penalizadas no ajustadas mostraron asociaciones de la clase Poly App Use con un alza en el número de parejas receptivas en relaciones sexuales anales y con reportes de sexo anal receptivo sin condón. Al ajustar por otras covariables, los miembros de la clase Poly App Use, con respecto a las otras clases, tuvieron más probabilidades de ser mayores (2534 vs. 1824, ORA = 3,81; IC95%=1,70 − 9,03), de informar el uso de drogas ilícitas en los últimos 6 meses (ORA = 2,93; IC95%=1,25 − 7,43) y haber utilizado alguna vez profilaxis-preexposición (ORA = 2,79; IC95%=1,10 − 7,12). Así mismo, los miembros de esta clase tuvieron perfiles de comportamiento asociado con el riesgo elevado de VIH y también informaron comportamientos de protección relacionados con el VIH que probablemente indican una mayor conciencia del riesgo entre ellos. Nuestros hallazgos justifican la diferenciación de perfiles de comportamiento en base a patrones de uso de aplicaciones y sugieren no generalizar los comportamientos sexuales asociados con riesgos potenciales de VIH a todos los HSH que las usan.
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Infecciones por VIH , Aplicaciones Móviles , Minorías Sexuales y de Género , Masculino , Humanos , Homosexualidad Masculina , Kentucky/epidemiología , Análisis de Clases Latentes , Infecciones por VIH/epidemiología , Infecciones por VIH/prevención & control , Conducta Sexual , Parejas SexualesRESUMEN
Life-cycle assessment (LCA) is one of the most widely applied methods for sustainability assessment. A main application of LCA is to compare alternative products to identify and promote those that are more environmentally friendly. Such comparative LCA studies often rest on, explicitly or implicitly, an idealized assumption, namely, 1:1 displacement between functionally equivalent products. However, product displacement in the real world is much more complicated, affected by various factors such as the rebound effect and policy schemes. Here, we quantitatively review studies that have considered these aspects to evaluate the magnitude and distribution of realistic displacement estimates across several major product categories (biofuels, electricity, electric vehicles, and recycled products). Results show that displacement ratios concentrate around 40-60%, suggesting considerable overestimation of the benefits of alternative products if the 1:1 displacement assumption was used. Overall, there have been a small number of modeling studies on realistic product displacement and their scopes were limited. Additional research is needed to cover more product categories and geographies and improve the modeling of market and policy complexities. As such research accumulates, their displacement estimates can form a database that can be drawn upon by comparative LCA studies to more accurately determine the environmental impacts of alternative products.
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Reciclaje , Biocombustibles , Modelos Teóricos , AmbienteRESUMEN
Reducing aviation emissions is important as they contribute to air pollution and climate change. Several alternative aviation fuels that may reduce life cycle emissions have been proposed. Comparative life cycle assessments (LCAs) of fuels are useful for inspecting individual fuels, but systemwide analysis remains difficult. Thus, systematic properties like fleet composition, performance, or emissions and changes to them under alternative fuels can only be partially addressed in LCAs. By integrating the geospatial fuel and emission model, AviTeam, with LCA, we can assess the mitigation potential of a fleetwide use of alternative aviation fuels on 210 000 shorter haul flights. In an optimistic case, liquid hydrogen (LH2) and power-to-liquid fuels, when produced with renewable electricity, may reduce emissions by about 950 GgCO2eq when assessed with the GWP100 metric and including non-CO2 impacts for all flights considered. Mitigation potentials range from 44% on shorter flights to 56% on longer flights. Alternative aviation fuels' mitigation potential is limited because of short-lived climate forcings and additional fuel demand to accommodate LH2 fuel. Our results highlight the importance of integrating system models into LCAs and are of value to researchers and decision-makers engaged in climate change mitigation in the aviation and transport sectors.
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Aviación , Emisiones de Vehículos , Modelos Teóricos , Contaminación del Aire , Cambio Climático , Contaminantes Atmosféricos/análisisRESUMEN
Resource recovery from wet organic wastes can support circular economies by creating financial incentives to produce renewable energy and return nutrients to agriculture. In this study, we characterize the potential for hydrothermal liquefaction (HTL)-based resource recovery systems to advance the economic and environmental sustainability of wastewater sludge, FOG (fats, oils, and grease), food waste, green waste, and animal manure management through the production of liquid biofuels (naphtha, diesel), fertilizers (struvite, ammonium sulfate), and power (heat, electricity). From the waste management perspective, median costs range from -193 $·tonne-1 (FOG) to 251 $·tonne-1 (green waste), and median carbon intensities range from 367 kg CO2 eq·tonne-1 (wastewater sludge) to 769 kg CO2 eq·tonne-1 (green waste). From the fuel production perspective, the minimum selling price of renewable diesel blendstocks are within the commercial diesel price range (2.37 to 5.81 $·gal-1) and have a lower carbon intensity than petroleum diesel (101 kg CO2 eq·MMBTU-1). Finally, through uncertainty analysis and Monte Carlo filtering, we set specific targets (i.e., achieve wastewater sludge-to-biocrude yield >0.440) for the future development of hydrothermal waste management system components. Overall, our work demonstrates the potential of HTL-based resource recovery systems to reduce the costs and carbon intensity of resource-rich organic wastes.
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Eliminación de Residuos , Aguas Residuales , Animales , Aguas del Alcantarillado , Dióxido de Carbono/análisis , Alimentos , Biocombustibles/análisis , CarbonoRESUMEN
OBJECTIVES: A mass spectrometry (LCâMS/MS)-based interlaboratory comparison study was performed for nine steroid analytes with five participating laboratories. The sample set contained 40 pooled samples of human serum generated from preanalyzed leftovers. To obtain a well-balanced distribution across reference intervals of each steroid, the leftovers first underwent a targeted mixing step. METHODS: All participants measured a sample set once using their own multianalyte protocols and calibrators. Four participants used in-house developed measurement platforms, including IVD-CE certified calibrators, which were used by three participants; the 5th lab used the whole LCâMS kit from an IVD manufacturer. All labs reported results for 17-hydroxyprogesterone, androstenedione, cortisol, and testosterone, and four labs reported results for 11-deoxycortisol, corticosterone, cortisone, dehydroepiandrosterone sulfate (DHEAS), and progesterone. RESULTS: Good or acceptable overall comparability was found in BlandâAltman and PassingâBablok analyses. Mean bias against the overall mean remained less than ±10â¯% except for DHEAS, androstenedione, and progesterone at one site and for cortisol and corticosterone at two sites (max. -18.9â¯% for androstenedione). The main analytical problems unraveled by this study included a bias not previously identified in proficiency testing, operator errors, non-supported matrix types and higher inaccuracy and imprecision at lower ends of measuring intervals. CONCLUSIONS: This study shows that intermethod comparison is essential for monitoring the validity of an assay and should serve as an example of how external quality assessment could work in addition to organized proficiency testing schemes.
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Hidrocortisona , Progesterona , Humanos , Cromatografía Liquida/métodos , Cromatografía Líquida con Espectrometría de Masas , Corticosterona , Androstenodiona , Espectrometría de Masas en Tándem/métodos , Esteroides , TestosteronaRESUMEN
A certified reference material (CRM, KRISS 108-01-002) for zearalenone in corn flour was developed to assure reliable and accurate measurements in testing laboratories. Commercially available corn flour underwent freeze-drying, pulverization, sieving, and homogenization. The final product was packed in amber bottles, approximately 14 g per unit, and preserved at -70 °C. 13C18-Zearalenone was used as an internal standard (IS) for the certification of zearalenone by isotope-dilution liquid chromatography-tandem mass spectrometry (ID-LCâMS/MS) and for the analysis of α-zearalenol, ß-zearalenol, and zearalanone by LCâMS/MS. The prepared CRM was sufficiently homogeneous, as the among-unit relative standard deviation for each mycotoxin ranged from 2.2 to 5.7 %. Additionally, the stability of the mycotoxins in the CRM was evaluated under different temperature conditions and scheduled test periods, including storage at -70°C, -20°C, and 4°C and room temperature for up to 12 months, 6 months, and 1 month, respectively. The content of each target mycotoxin in the CRM remained stable throughout the monitoring period at each temperature. Zearalenone content (153.6 ± 8.0 µg/kg) was assigned as the certified value. Meanwhile, the contents of α-zearalenol (1.30 ± 0.17 µg/kg), ß-zearalenol (4.75 ± 0.33 µg/kg), and zearalanone (2.09 ± 0.16 µg/kg) were provided as informative values.
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Harina , Estándares de Referencia , Espectrometría de Masas en Tándem , Zea mays , Zearalenona , Zearalenona/análisis , Zea mays/química , Harina/análisis , Harina/normas , Espectrometría de Masas en Tándem/métodos , Cromatografía Liquida/métodos , Límite de Detección , Contaminación de Alimentos/análisis , Reproducibilidad de los ResultadosRESUMEN
Microalgae is considered as sustainable and viable feedstock for biofuel production due to its significant advantages over terrestrial plants. Algal biofuels have received significant attention among researchers and energy experts owing to an upsurge in global energy issues emanating from depletion in fossil fuel reserves increasing greenhouse gases emission conflict among agricultural crops, traditional biomass feedstock, and potential futuristic energy security. Further, the exploration of value-added microalgae as sustainable and viable feedstock for the production of variety of biofuels such as biogas, bio-hydrogen, bioethanol, and biodiesel are addressed. Moreover, the assessment of life-cycle, energy balance, and environmental impacts of biofuel production from microalgae are briefly discussed. The present study focused on recent advancements in synthetic biology, metabolic engineering tools, algal bio refinery, and the optimization of algae growth conditions. This paper also elucidates the function of microalgae as bio refineries, the conditions of algae-based cultures, and other operational factors that must be adjusted to produce biofuels that are price-competitive with fossil fuels.
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BACKGROUND: Micro- and nanoplastics (MNPs) represent one of the most widespread environmental pollutants of the twenty-first century to which all humans are orally exposed. Upon ingestion, MNPs pass harsh biochemical conditions within the gastrointestinal tract, causing a unique protein corona on the MNP surface. Little is known about the digestion-associated protein corona and its impact on the cellular uptake of MNPs. Here, we systematically studied the influence of gastrointestinal digestion on the cellular uptake of neutral and charged polystyrene MNPs using THP-1-derived macrophages. RESULTS: The protein corona composition was quantified using LCâMS-MS-based proteomics, and the cellular uptake of MNPs was determined using flow cytometry and confocal microscopy. Gastrointestinal digestion resulted in a distinct protein corona on MNPs that was retained in serum-containing cell culture medium. Digestion increased the uptake of uncharged MNPs below 500 nm by 4.0-6.1-fold but did not affect the uptake of larger sized or charged MNPs. Forty proteins showed a good correlation between protein abundance and MNP uptake, including coagulation factors, apolipoproteins and vitronectin. CONCLUSION: This study provides quantitative data on the presence of gastrointestinal proteins on MNPs and relates this to cellular uptake, underpinning the need to include the protein corona in hazard assessment of MNPs.
Asunto(s)
Microplásticos , Corona de Proteínas , Humanos , Microplásticos/toxicidad , Corona de Proteínas/química , Corona de Proteínas/metabolismo , Poliestirenos/toxicidad , Plásticos , DigestiónRESUMEN
PURPOSE: Leber congenital amaurosis (LCA) is a group of early-onset retinal degenerative disorders, resulting in blindness in children. This study aimed to describe the clinical and genetic characteristics of a cohort of patients with LCA and to investigate the retinal vascular characteristics in LCA patients. METHODS: Fifty-two children with LCA were included in the study. All patients underwent detailed ocular examinations. Electroretinography (ERG) was used to evaluate the retinal function. Optical coherence tomography (OCT) was used to assess the structure change of the retina for those patients who were able to cooperate very well. Panel-based next-generation sequencing was performed to identify pathogenic variants in genes associated with LCA. Diameters of the retinal vessels were measured using the EVision AI screening system with an artificial intelligence (AI) technique. An ultrasound Doppler was used to evaluate hemodynamic parameters, including peak systolic velocity (PSV), resistive index (RI), and pulsatility index (PI), in the ophthalmic, central retinal, posterior ciliary, carotid, and internal carotid as well as external carotid arteries in 12 patients aged from 3 to 14 years. RESULTS: We detected 75 pathogenic variants from ten genes of RPGRIP1, CEP290, GUCY2D, LCA5, AIPL1, CRB1, RPE65, CRX, RDH12, and TULP1, including 29 novel and 36 previously reported variants in 52 affected children with LCA, with the highest detective rate in RPGRIP1 (26.9%). Fundus appearance is diverse in patients with LCA, ranging from normal to severe peripheral or central retinopathy. Retinal vasculature was evaluated in 12 patients with different gene variants, showing narrowed arteries with an average diameter of 43.6 ± 3.8 µm compared to that of 51.7 ± 2.6 µm in the normal controls (P < 0.001, n = 12). Meanwhile, their hemodynamic parameters were changed as well in the ophthalmic artery (OA), with a decreased PSV (P = 0.0132, n = 12) and slightly increased PI (P = 0.0488, n = 12) compared to the normal controls. However, the hemodynamic parameters did not change significantly in the other vessels. CONCLUSIONS: Blood supply to the eyeball is predicted to be reduced in patients with LCA, presumably due to photoreceptor cell degeneration. The novel identified variants will expand the spectrum of variants in LCA-related genes and be useful for studying the molecular mechanisms of LCA.
Asunto(s)
Electrorretinografía , Amaurosis Congénita de Leber , Mutación , Tomografía de Coherencia Óptica , Humanos , Amaurosis Congénita de Leber/genética , Amaurosis Congénita de Leber/fisiopatología , Amaurosis Congénita de Leber/diagnóstico , Masculino , Niño , Femenino , Tomografía de Coherencia Óptica/métodos , Adolescente , Preescolar , Vasos Retinianos , Proteínas del Ojo/genética , Agudeza Visual , ADN/genética , Análisis Mutacional de ADN , Retina/fisiopatologíaRESUMEN
OBJECTIVES: Dietary environmental impact in a Norwegian adult population was estimated for six environmental impact categories. Moreover, environmental benefits of scenario diets complying with the Norwegian Food-Based Dietary Guidelines (FBDG) and the EAT-Lancet reference diet were assessed. DESIGN: The current diet of Norwegian adults was estimated according to 24-h dietary recall data from a national dietary surveillance survey (Norkost 3). Scenario diets were modelled to represent the Norwegian FBDG and the EAT-Lancet healthy reference diet. Dietary environmental impact in terms of global warming potential, freshwater and marine eutrophication, terrestrial acidification, water use and transformation and use of land was estimated for the current and scenario diets using environmental impact data representative of the Norwegian market. Significant associations between impact and gender/educational attainment were assessed at P < 0·05. SETTING: Norway. PARTICIPANTS: Adults (n=1787) aged 18-70 years who participated in the Norkost 3 survey (2010-2011). RESULTS: Environmental impact varied significantly by gender and educational attainment. The food groups contributing most to environmental impact of Norwegian diets were meat, dairy, beverages, grains and composite dishes. Compared with the current Norwegian diet, the FBDG scenario reduced impacts from 2 % (freshwater eutrophication) to 32 % (water use), while the EAT-Lancet scenario reduced impacts from 7 % (marine eutrophication) to 61 % (land use). The EAT-Lancet scenario resulted in 3-48 % larger reductions in impact than the FBDG scenario. CONCLUSIONS: The Norwegian FBDG, while not as environmentally friendly as the EAT-Lancet reference diet, can still be an important tool in lessening environmental burden of Norwegian diets.