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1.
Bioresour Technol ; 410: 131214, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39127361

RESUMEN

Despite its prominence, the ability to engineer Cupriavidus necator H16 for inorganic carbon uptake and fixation is underexplored. We tested the roles of endogenous and heterologous genes on C. necator inorganic carbon metabolism. Deletion of ß-carbonic anhydrase can had the most deleterious effect on C. necator autotrophic growth. Replacement of this native uptake system with several classes of dissolved inorganic carbon (DIC) transporters from Cyanobacteria and chemolithoautotrophic bacteria recovered autotrophic growth and supported higher cell densities compared to wild-type (WT) C. necator in batch culture. Strains expressing Halothiobacillus neopolitanus DAB2 (hnDAB2) and diverse rubisco homologs grew in CO2 similarly to the wild-type strain. Our experiments suggest that the primary role of carbonic anhydrase during autotrophic growth is to support anaplerotic metabolism, and an array of DIC transporters can complement this function. This work demonstrates flexibility in HCO3- uptake and CO2 fixation in C. necator, providing new pathways for CO2-based biomanufacturing.


Asunto(s)
Dióxido de Carbono , Cupriavidus necator , Dióxido de Carbono/metabolismo , Cupriavidus necator/metabolismo , Cupriavidus necator/genética , Bicarbonatos/metabolismo , Ciclo del Carbono/fisiología , Anhidrasas Carbónicas/metabolismo , Procesos Autotróficos , Halothiobacillus/metabolismo , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Ribulosa-Bifosfato Carboxilasa/metabolismo
2.
Microb Biotechnol ; 17(9): e70003, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-39215388

RESUMEN

The exploration of novel hosts with the ability to assimilate formic acid, a C1 substrate that can be produced from renewable electrons and CO2, is of great relevance for developing novel and sustainable biomanufacturing platforms. Formatotrophs can use formic acid or formate as a carbon and/or reducing power source. Formatotrophy has typically been studied in neutrophilic microorganisms because formic acid toxicity increases in acidic environments below the pKa of 3.75 (25°C). Because of this toxicity challenge, utilization of formic acid as either a carbon or energy source has been largely unexplored in thermoacidophiles, species that possess the ability to produce a variety of metabolites and enzymes of high biotechnological relevance. Here we investigate the capacity of several thermoacidophilic archaea species from the Sulfolobales order to tolerate and metabolize formic acid. Metallosphaera prunae, Sulfolobus metallicus and Sulfolobus acidocaldarium were found to metabolize and grow with 1-2 mM of formic acid in batch cultivations. Formic acid was co-utilized by this species alongside physiological electron donors, including ferrous iron. To enhance formic acid utilization while maintaining aqueous concentrations below the toxicity threshold, we developed a bioreactor culturing method based on a sequential formic acid feeding strategy. By dosing small amounts of formic acid sequentially and feeding H2 as co-substrate, M. prunae could utilize a total of 16.3 mM of formic acid and grow to higher cell densities than when H2 was supplied as a sole electron donor. These results demonstrate the viability of culturing thermoacidophilic species with formic acid as an auxiliary substrate in bioreactors to obtain higher cell densities than those yielded by conventional autotrophic conditions. Our work underscores the significance of formic acid metabolism in extreme habitats and holds promise for biotechnological applications in the realm of sustainable energy production and environmental remediation.


Asunto(s)
Formiatos , Formiatos/metabolismo , Sulfolobales/metabolismo , Sulfolobales/genética
3.
Foods ; 13(7)2024 Mar 25.
Artículo en Inglés | MEDLINE | ID: mdl-38611309

RESUMEN

Opuntia ficus-indica var. Colorada fruit is an important source of indicaxanthin, a betalain with antioxidant, anti-inflammatory, and neuromodulatory potential, proven in both in vitro and in vivo models. Other betalains and phenolic compounds with bioactive activities have also been identified in Colorada fruit extracts. These compounds may degrade by their exposure to different environmental factors, so in the present research, two double emulsion systems (W1/O/W2) were elaborated using Tween 20 (TW) and sodium caseinate (SC) as surfactants to encapsulate Colorada fruit pulp extracts, with the aim of enhancing their stability during storage. Encapsulation efficiencies of up to 97.3 ± 2.7%, particle sizes between 236 ± 4 and 3373 ± 64 nm, and zeta potential values of up to ∣46.2∣ ± 0.3 mV were obtained. In addition, the evaluation of the in vitro gastro-intestinal stability and bioaccessibility of the main individual bioactives was carried out by standardized INFOGEST© protocol, obtaining the highest values for the encapsulated extract bioactives in comparison with the non-encapsulated extract (control). Especially, TW double emulsion showed bioaccessibility values of up to 82.8 ± 1.5% for the main bioactives (indicaxanthin, piscidic acid, and isorhamnetin glucoxyl-rhamnosyl-pentoside 2 (IG2)), indicating a promising potential for its use as a functional natural colorant ingredient.

4.
Foods ; 13(8)2024 Apr 18.
Artículo en Inglés | MEDLINE | ID: mdl-38672912

RESUMEN

Plant-based beverages are one of the foods that currently arouse a lot of interest in the population due to their composition with compounds beneficial to health in addition to their being used as milk substitutes for people who suffer from food disorders. Also, their fortification with different nutrients or healthy ingredients with the aim of improving plant-based health potential is actually gaining importance in the food industry. For this reason, the aim of the present investigation was the preparation of a healthy Brazil nut beverage enriched with Opuntia stricta var. dillenii pulp green extracts (ODPs), in order to produce a healthy plant-based beverage with improved nutritional characteristics. The microstructural characterization of the Brazil nut beverage, its stability during cold storage for up to 24 days at 5 °C, the composition of bioactive compounds provided via ODP extract (betalains and phenolic compounds), and their antioxidant activity were evaluated in this study. Green ODP extracts (0.5 and 1 g/100 g beverage) were added to a standardized Brazil nut beverage (reduced fat beverage). The characterization of the bioactive composition (betalains and phenolic compounds) of the elaborated beverage was achieved via HPLC (UV-vis and MS-QT of detection), and the antioxidant activity measurements via ORAC were also carried out. Optical microscopy, particle size, and Z potential analysis was conducted to characterize the structure of the Brazil nut beverages as food emulsions in which ODP extract was added. Most of the bioactive compounds from the green ODP extract added to the beverages showed good retention and remained stable throughout the 24 days of storage at 7 °C, with encapsulation efficiencies ranging from 98.34% to 92.35% for betalains and from 93.67% and 81.20% for phenolic compounds. According to the results of this study, Brazil nut beverage seems to be a healthy and efficient food emulsion system to encapsulate ODP extract rich in betalains and phenolic compounds, with high antioxidant activity, making possible the development of a Brazil nut beverage with improved health potential.

6.
Microb Biotechnol ; 17(1): e14383, 2024 01.
Artículo en Inglés | MEDLINE | ID: mdl-38231155

RESUMEN

Microbial electrosynthesis (MES) constitutes a bioelectrochemical process where bacteria uptake electrons extracellularly from a polarized electrode to incorporate them into their anabolic metabolism. However, the efficiency of current MES reactor designs can be lower than expected due to limitations regarding electron transfer and mass transport. One of the most promising bioreactor configurations to overcome these bottlenecks is the Microbial Electrochemical Fluidized Bed Reactor (ME-FBR). In this study, microbial CO2 fixation is investigated for the first time in a ME-FBR operated as a 3-phase reactor (solid-liquid-gas). An electroconductive carbon bed, acting as a working electrode, was fluidized with gas and polarized at different potentials (-0.6, -0.8 and -1 V vs. Ag/AgCl) so it could act as an electron donor (biocathode). Under these potentials, CO2 fixation and electron transfer were evaluated. Autotrophic electroactive microorganisms from anaerobic wastewater were enriched in a ME-FBR in the presence of 2-bromoethanosulfonic acid (BES) to inhibit the growth of methanogens. Cyclic voltammetry analysis revealed interaction between the microorganisms and the cathode. Furthermore, volatile fatty acids like propionate, formate and acetate were detected in the culture supernatant. Acetate production had a maximum rate of ca. 1 g L-1 day-1 . Planktonic cell biomass was produced under continuous culture at values as high as ca. 0.7 g L-1 dry weight. Overall, this study demonstrates the feasibility of employing a fluidized electrode with gaseous substrates and electricity as the energy source for generating biomass and carboxylic acids.


Asunto(s)
Dióxido de Carbono , Ácidos Grasos Volátiles , Dióxido de Carbono/metabolismo , Biomasa , Acetatos , Electrodos
7.
Hum Immunol ; 85(2): 110749, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38238229

RESUMEN

BACKGROUND: Alloantibodies, especially anti-human leukocyte antigen antibodies (HLA antibodies), and autoantibodies, as angiotensin II type 1 receptor antibodies (AT1R antibodies), may complicate the access and the course of transplantation. Pregnancy is a known source of HLA antibodies, with most studies evaluating pregnancy-induced sensitization by complement-dependent cytotoxicity assays, mainly after childbirth. AT1R antibodies have been evaluated in the context of preeclampsia. We aimed to evaluate pregnancy as a natural source of HLA antibodies and AT1R antibodies, their dynamics along gestation and the potential factors involved in antibody appearance. METHODS: Serum samples from pregnant women were collected during the three trimesters of pregnancy (1T, 2T, 3T). Presence of HLA antibodies was assessed by screening beads on Luminex and AT1R antibodies by ELISA. RESULTS: A cohort of 138 pregnant women were included. Samples from all were tested in 1T, 127 in 2T and 102 in 3T. HLA antibodies increased from 29.7 % (1T) to 38.2 % (3T). AT1R antibodies were stable around 30 % along pregnancy. Up to 43.2 % multiparous women had HLA antibodies, with a similar proportion of class I and class II antibodies. In primiparous women HLA antibodies increased along pregnancy (from 17.6 % to 34.1 %), with predominance of class II HLA antibodies. AT1R antibodies were not different in primiparous and multiparous women. CONCLUSIONS: Pregnancy is a relevant source of HLA antibodies sensitization, but not of AT1R antibodies. HLA antibodies increased clearly in primiparous women with predominance of class II. The use of newer solid-phase techniques on Luminex evidence a higher degree of HLA sensitization during pregnancy.


Asunto(s)
Trasplante de Riñón , Humanos , Femenino , Embarazo , Receptor de Angiotensina Tipo 1 , Rechazo de Injerto , Autoanticuerpos , Antígenos HLA
9.
An. pediatr. (2003. Ed. impr.) ; 99(6): 393-402, Dic. 2023. graf, tab, mapas
Artículo en Inglés, Español | IBECS | ID: ibc-228662

RESUMEN

Introducción: La principal medida de prevención frente a la enfermedad meningocócica invasiva es la vacunación. El objetivo de este estudio es evaluar la aceptabilidad y las desigualdades socioeconómicas en el acceso a la vacuna frente a meningococo B (MenB) en la Comunidad de Madrid en el periodo previo a la introducción de la misma en el calendario. Materiales y métodos: Se realizó un estudio observacional descriptivo en la cohorte de niños/as nacidos entre 2016 y 2019, de tipo ecológico, empleando registros poblacionales electrónicos. Se describieron las coberturas de vacunación, se analizaron los factores asociados al estado vacunal, se describieron las distribuciones espaciales de cobertura de vacunación y de índice de privación (IP) y se analizó la asociación entre ambas mediante regresión espacial. Resultados: Se observó una tendencia creciente de las coberturas de primovacunación, pasando de un 44% en la cohorte de nacidos en el año 2016 a un 68% en la cohorte de 2019. Se encontró asociación estadísticamente significativa entre el estado vacunal y el IP (OR de primovacunación en zonas con IP5 respecto a zonas con IP1: 0,38; IC 95%: 0,39-0,50; p<0,001). El análisis espacial mostró correlación inversa entre el IP y la cobertura de vacunación. Conclusiones: El ascenso de las coberturas de esta vacuna muestra aceptación por parte de la población. La relación entre nivel socioeconómico y cobertura de vacunación confirma la existencia de una desigualdad en salud, y subraya la importancia de su inclusión en el calendario.(AU)


Introduction: The main preventive measure against invasive meningococcal disease is vaccination. The aim of our study was to evaluate the acceptability of the meningococcal B (MenB) vaccine and socioeconomic inequalities in the access to the vaccine in the Community of Madrid in the period prior to its introduction in the immunization schedule. Materials and methods: We conducted an observational and ecological descriptive study in the cohort of children born between 2016 and 2019 using population-based electronic records. We calculated the vaccination coverage and analysed factors associated with vaccination status, determined the spatial distribution of vaccination coverage and the deprivation index (DI) and assessed the association between them by means of spatial regression. Results: We observed an increasing trend in primary vaccination coverage, from 44% in the cohort born in 2016 to 68% in the 2019 cohort. We found a statistically significant association between vaccination status and the DI (OR of primary vaccination in areas with DI5 compared to areas with DP1, 0.38; 95% confidence interval: 0.39-0.50; P<.001). The spatial analysis showed an inverse correlation between the DI and vaccination coverage. Conclusions: The rise in the coverages of the MenB vaccine shows acceptance by the population. The association between socioeconomic level and vaccination coverage confirms the existence of health inequality and underlines the importance including this vaccine in the immunization schedule.(AU)


Asunto(s)
Humanos , Masculino , Femenino , Recién Nacido , Niño , Neisseria meningitidis Serogrupo B/inmunología , Meningitis Meningocócica/inmunología , Cobertura de Vacunación , Infecciones Meningocócicas/inmunología , España , Estudios de Cohortes , Epidemiología Descriptiva , Meningitis Meningocócica/prevención & control , Vacunación , Infecciones Meningocócicas/prevención & control
10.
Front Microbiol ; 14: 1298023, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38075918

RESUMEN

A subset of microorganisms that perform respiration can endogenously utilize insoluble electron donors, such as Fe(II) or a cathode, in a process called extracellular electron transfer (EET). However, it is unknown whether similar endogenous EET can be performed by primarily fermentative species like lactic acid bacteria. We report for the first time electron uptake from a cathode by Lactiplantibacillus plantarum, a primarily fermentative bacteria found in the gut of mammals and in fermented foods. L. plantarum consumed electrons from a cathode and coupled this oxidation to the reduction of both an endogenous organic (pyruvate) and an exogenous inorganic electron acceptor (nitrate). This electron uptake from a cathode reroutes glucose fermentation toward lactate degradation and provides cells with a higher viability upon sugar exhaustion. Moreover, the associated genes and cofactors indicate that this activity is mechanistically different from that one employed by lactic acid bacteria to reduce an anode and to perform respiration. Our results expand our knowledge of the diversity of electroactive species and of the metabolic and bioenergetic strategies used by lactic acid bacteria.

11.
mBio ; : e0223423, 2023 Nov 20.
Artículo en Inglés | MEDLINE | ID: mdl-37982640

RESUMEN

IMPORTANCE: While quinones are essential for respiratory microorganisms, their importance for microbes that rely on fermentation metabolism is not understood. This gap in knowledge hinders our understanding of anaerobic microbial habitats, such in mammalian digestive tracts and fermented foods. We show that Lactiplantibacillus plantarum, a model fermentative lactic acid bacteria species abundant in human, animal, and insect microbiomes and fermented foods, uses multiple exogenous, environmental quinones as electron shuttles for a hybrid metabolism involving EET. Interestingly, quinones both stimulate this metabolism as well as cause oxidative stress when extracellular electron acceptors are absent. We also found that quinone-producing, lactic acid bacteria species commonly enriched together with L. plantarum in food fermentations accelerate L. plantarum growth and medium acidification through a mainly quinone- and EET-dependent mechanism. Thus, our work provides evidence of quinone cross-feeding as a key ecological feature of anaerobic microbial habitats.

12.
Prog Brain Res ; 282: 95-121, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38035911

RESUMEN

Numerical cognition is an essential skill for survival, which includes the processing of discrete and continuous quantities, involving a mainly right fronto-parietal network. However, the neurocognitive systems underlying the processing and integration of discrete and continuous quantities are currently under debate. Noninvasive brain stimulation techniques have been used in the study of the neural basis of numerical cognition with a spatial, temporal and functional resolution superior to other neuroimaging techniques. The present randomized sham-controlled single-blinded trial addresses the involvement of the right dorsolateral prefrontal cortex and the right intraparietal sulcus in magnitude processing and integration. Multifocal anodal transcranial direct current stimulation was applied online during the execution of magnitude comparison tasks in three conditions: right prefrontal, right parietal and sham stimulation. The results show that prefrontal stimulation produced a moderated decrease in response times in all magnitude processing and integration tasks compared to sham condition. While parietal stimulation had no significant effect on any of the tasks. The effect found is interpreted as a generalized improvement in processing speed and magnitude integration due to right prefrontal neuromodulation, which may be attributable to domain-general or domain-specific factors.


Asunto(s)
Estimulación Transcraneal de Corriente Directa , Humanos , Cognición/fisiología , Corteza Prefrontal/fisiología , Velocidad de Procesamiento
13.
An Pediatr (Engl Ed) ; 99(6): 393-402, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38016859

RESUMEN

INTRODUCTION: The main preventive measure against invasive meningococcal disease is vaccination. The aim of our study was to evaluate the acceptability of the meningococcal B (MenB) vaccine and socioeconomic inequalities in the access to the vaccine in the Community of Madrid in the period prior to its introduction in the immunization schedule. MATERIALS AND METHODS: We conducted an observational and ecological descriptive study in the cohort of children born between 2016 and 2019 using population-based electronic records. We calculated the vaccination coverage and analysed factors associated with vaccination status, determined the spatial distribution of vaccination coverage and the deprivation index (DI) and assessed the association between them by means of spatial regression. RESULTS: We observed an increasing trend in primary vaccination coverage, from 44% in the cohort born in 2016 to 68% in the 2019 cohort. We found a statistically significant association between vaccination status and the DI (OR of primary vaccination in areas with DI5 compared to areas with DP1, 0.38; 95% confidence interval, 0.39-0.50; P<.001). The spatial analysis showed an inverse correlation between the DI and vaccination coverage. CONCLUSIONS: The rise in the coverages of the MenB vaccine shows acceptance by the population. The association between socioeconomic level and vaccination coverage confirms the existence of health inequality and underlines the importance including this vaccine in the immunization schedule.


Asunto(s)
Disparidades en el Estado de Salud , Vacunas Meningococicas , Niño , Humanos , Esquemas de Inmunización , Vacunación , Factores Socioeconómicos
14.
Eur J Hum Genet ; 31(10): 1117-1124, 2023 10.
Artículo en Inglés | MEDLINE | ID: mdl-37500725

RESUMEN

Nuclear receptor subfamily 2 group F member 2 (NR2F2 or COUP-TF2) encodes a transcription factor which is expressed at high levels during mammalian development. Rare heterozygous Mendelian variants in NR2F2 were initially identified in individuals with congenital heart disease (CHD), then subsequently in cohorts of congenital diaphragmatic hernia (CDH) and 46,XX ovotesticular disorders/differences of sexual development (DSD); however, the phenotypic spectrum associated with pathogenic variants in NR2F2 remains poorly characterized. Currently, less than 40 individuals with heterozygous pathogenic variants in NR2F2 have been reported. Here, we review the clinical and molecular details of 17 previously unreported individuals with rare heterozygous NR2F2 variants, the majority of which were de novo. Clinical features were variable, including intrauterine growth restriction (IUGR), CHD, CDH, genital anomalies, DSD, developmental delays, hypotonia, feeding difficulties, failure to thrive, congenital and acquired microcephaly, dysmorphic facial features, renal failure, hearing loss, strabismus, asplenia, and vascular malformations, thus expanding the phenotypic spectrum associated with NR2F2 variants. The variants seen were predicted loss of function, including a nonsense variant inherited from a mildly affected mosaic mother, missense and a large deletion including the NR2F2 gene. Our study presents evidence for rare, heterozygous NR2F2 variants causing a highly variable syndrome of congenital anomalies, commonly associated with heart defects, developmental delays/intellectual disability, dysmorphic features, feeding difficulties, hypotonia, and genital anomalies. Based on the new and previous cases, we provide clinical recommendations for evaluating individuals diagnosed with an NR2F2-associated disorder.


Asunto(s)
Anomalías Múltiples , Cardiopatías Congénitas , Hernias Diafragmáticas Congénitas , Discapacidad Intelectual , Animales , Humanos , Anomalías Múltiples/genética , Anomalías Múltiples/diagnóstico , Factor de Transcripción COUP II/genética , Cardiopatías Congénitas/genética , Hernias Diafragmáticas Congénitas/genética , Discapacidad Intelectual/genética , Hipotonía Muscular , Síndrome
15.
Foods ; 12(11)2023 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-37297486

RESUMEN

Opuntia cactus fruit (prickly pear flesh and agricultural residues such as peels and stalks) is an important source of bioactive compounds, including betalains and phenolic compounds. In this work, two double emulsion W1/O/W2 formulations (A and B) were designed to encapsulate green extracts rich in betalains and phenolic compounds obtained from Opuntia stricta var. dillenii (OPD) fruits with the aim of improving their stability and protecting them during the in vitro gastrointestinal digestion process. In addition, the characterization of the double emulsions was studied by microscopy and the evaluation of their physical and physico-chemical parameters. Formulation A, based on Tween 20, showed smaller droplets (1.75 µm) and a higher physical stability than Formulation B, which was achieved with sodium caseinate (29.03 µm). Regarding the encapsulation efficiency of the individual bioactives, betalains showed the highest values (73.7 ± 6.7 to 96.9 ± 3.3%), followed by flavonoids (68.2 ± 5.9 to 95.9 ± 7.7%) and piscidic acid (71 ± 1.3 to 70.2 ± 5.7%) depending on the formulation and the bioactive compound. In vitro digestive stability and bioaccessibility of the individual bioactives increased when extracts were encapsulated for both formulations (67.1 to 253.1%) in comparison with the non-encapsulated ones (30.1 to 64.3%), except for neobetanin. Both formulations could be considered as appropriate microcarrier systems for green OPD extracts, especially formulation A. Further studies need to be conducted about the incorporation of these formulations to develop healthier foods.

16.
Dermatol Online J ; 29(6)2023 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-38478665

RESUMEN

A 9-year-old boy diagnosed with acute myeloblastic leukemia and undergoing chemotherapy, was admitted with febrile neutropenia. During his admission, several violaceous plaques appeared on the upper extremities and anterior left hemithorax, which worsened and acquired a necrotic center. We performed a biopsy and histology showed a cutaneous infarction at the dermoepidermal and subcutaneous level. We observed abundant wide hyphae with right-angled branching and a culture isolated Rhizopus oryzae. A plastic surgery consultant performed a surgical debridement of the lesions and treatment was started with intravenous amphotericin B. The patient did well on treatment and after almost a month of hospitalization, he was discharged with oral posaconazole. Mucormycosis is an opportunistic fungal infection associated with immunosuppression, particularly involving prematurity and hematological diseases in the pediatric age group. Multiple lesions, as in our case, are infrequent. The clinical presentation is variable. Direct smear or histological observation is the quickest diagnostic technique whereas culture is the most definitive. The combination of surgical debridement and amphotericin B is the treatment with the highest survival rates.


Asunto(s)
Anfotericina B , Mucormicosis , Masculino , Humanos , Niño , Anfotericina B/uso terapéutico , Rhizopus , Mucormicosis/diagnóstico , Mucormicosis/terapia , Piel/patología , Necrosis/complicaciones , Antifúngicos/uso terapéutico
17.
Front Endocrinol (Lausanne) ; 13: 957969, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36303863

RESUMEN

The palmitoylation of the Hedgehog (Hh) family of morphogens, named sonic hedgehog (SHH), desert hedgehog (DHH), and Indian hedgehog (IHH), is crucial for effective short- and long-range signaling. The hedgehog acyltransferase (HHAT) attaches the palmitate molecule to the Hh; therefore, variants in HHAT cause a broad spectrum of phenotypes. A missense HHAT novel variant c.1001T>A/p.(Met334Lys) was described in a patient first referred for a 46,XY different sexual development with partial gonadal dysgenesis but with microcephaly, eye defects, and distal phalangeal hypoplasia of both thumbs. The in silico analysis of the variant predicted an affectation of the nearest splicing site. Thus, in vitro minigene studies were carried out, which demonstrated that the variant does not affect the splicing. Subsequent protein in silico studies supported the pathogenicity of the variant, and, in conclusion, this was considered the cause of the patient's phenotype.


Asunto(s)
Disgenesia Gonadal , Microcefalia , Humanos , Proteínas Hedgehog/genética , Proteínas Hedgehog/metabolismo , Microcefalia/complicaciones , Microcefalia/genética , Pulgar , Desarrollo Sexual , Aciltransferasas/genética
18.
Elife ; 112022 04 05.
Artículo en Inglés | MEDLINE | ID: mdl-35380108

RESUMEN

Cellular respiration is essential for multiple bacterial pathogens and a validated antibiotic target. In addition to driving oxidative phosphorylation, bacterial respiration has a variety of ancillary functions that obscure its contribution to pathogenesis. We find here that the intracellular pathogen Listeria monocytogenes encodes two respiratory pathways which are partially functionally redundant and indispensable for pathogenesis. Loss of respiration decreased NAD+ regeneration, but this could be specifically reversed by heterologous expression of a water-forming NADH oxidase (NOX). NOX expression fully rescued intracellular growth defects and increased L. monocytogenes loads >1000-fold in a mouse infection model. Consistent with NAD+ regeneration maintaining L. monocytogenes viability and enabling immune evasion, a respiration-deficient strain exhibited elevated bacteriolysis within the host cytosol and NOX expression rescued this phenotype. These studies show that NAD+ regeneration represents a major role of L. monocytogenes respiration and highlight the nuanced relationship between bacterial metabolism, physiology, and pathogenesis.


Cellular respiration is one of the main ways organisms make energy. It works by linking the oxidation of an electron donor (like sugar) to the reduction of an electron acceptor (like oxygen). Electrons pass between the two molecules along what is known as an 'electron transport chain'. This process generates a force that powers the production of adenosine triphosphate (ATP), a molecule that cells use to store energy. Respiration is a common way for cells to replenish their energy stores, but it is not the only way. A simpler process that does not require a separate electron acceptor or an electron transport chain is called fermentation. Many bacteria have the capacity to perform both respiration and fermentation and do so in a context-dependent manner. Research has shown that respiration can contribute to bacterial diseases, like tuberculosis and listeriosis (a disease caused by the foodborne pathogen Listeria monocytogenes). Indeed, some antibiotics even target bacterial respiration. Despite being often discussed in the context of generating ATP, respiration is also important for many other cellular processes, including maintaining the balance of reduced and oxidized nicotinamide adenine dinucleotide (NAD) cofactors. Because of these multiple functions, the exact role respiration plays in disease is unknown. To find out more, Rivera-Lugo, Deng et al. developed strains of the bacterial pathogen Listeria monocytogenes that lacked some of the genes used in respiration. The resulting bacteria were still able to produce energy, but they became much worse at infecting mammalian cells. The use of a genetic tool that restored the balance of reduced and oxidized NAD cofactors revived the ability of respiration-deficient L. monocytogenes to infect mammalian cells, indicating that this balance is what the bacterium requires to infect. Research into respiration tends to focus on its role in generating ATP. But these results show that for some bacteria, this might not be the most important part of the process. Understanding the other roles of respiration could change the way that researchers develop antibacterial drugs in the future. This in turn could help with the growing problem of antibiotic resistance.


Asunto(s)
Listeria monocytogenes , Listeriosis , Animales , Respiración de la Célula , Modelos Animales de Enfermedad , Evasión Inmune , Listeria monocytogenes/genética , Listeriosis/metabolismo , Listeriosis/microbiología , Listeriosis/patología , Ratones , NAD/metabolismo
19.
Elife ; 112022 02 11.
Artículo en Inglés | MEDLINE | ID: mdl-35147079

RESUMEN

Energy conservation in microorganisms is classically categorized into respiration and fermentation; however, recent work shows some species can use mixed or alternative bioenergetic strategies. We explored the use of extracellular electron transfer for energy conservation in diverse lactic acid bacteria (LAB), microorganisms that mainly rely on fermentative metabolism and are important in food fermentations. The LAB Lactiplantibacillus plantarum uses extracellular electron transfer to increase its NAD+/NADH ratio, generate more ATP through substrate-level phosphorylation, and accumulate biomass more rapidly. This novel, hybrid metabolism is dependent on a type-II NADH dehydrogenase (Ndh2) and conditionally requires a flavin-binding extracellular lipoprotein (PplA) under laboratory conditions. It confers increased fermentation product yield, metabolic flux, and environmental acidification in laboratory media and during kale juice fermentation. The discovery of a single pathway that simultaneously blends features of fermentation and respiration in a primarily fermentative microorganism expands our knowledge of energy conservation and provides immediate biotechnology applications.


Bacteria produce the energy they need to live through two processes, respiration and fermentation. While respiration is often more energetically efficient, many bacteria rely on fermentation as their sole means of energy production. Respiration normally depends on the presence of small soluble molecules, such as oxygen, that can diffuse inside the cell, but some bacteria can use metals or other insoluble compounds found outside the cell to perform 'extracellular electron transfer'. Lactic acid bacteria are a large group of bacteria that have several industrial uses and live in many natural environments. These bacteria survive using fermentation, but they also carry a group of genes needed for extracellular electron transfer. It is unclear whether they use these genes for respiration or if they have a different purpose. Tejedor-Sanz, Stevens et al. used a lactic acid bacterium called Lactiplantibacillus plantarum to study whether and how this group of bacteria use extracellular electron transfer. Analysis of L. plantarum and its effect on its surroundings showed that these bacteria use a hybrid process to produce energy: the cells use aspects of extracellular respiration to increase the yield and efficiency of fermentation. Combining these two approaches may allow L. plantarum to adapt to different environments and grow faster, allowing it to compete against other species. Tejedor-Sanz, Stevens et al. provide new information on a widespread group of bacteria that are often used in food production and industry. The next step will be to understand how the hybrid system is controlled and how it varies among species. Understanding this process could result in new biotechnologies and foods that are healthier, produce less waste, or have different tastes and textures.


Asunto(s)
Transporte de Electrón/fisiología , Fermentación , Lactobacillaceae/metabolismo , Albinismo Oculocutáneo , Biomasa , Brassica/química , Jugos de Frutas y Vegetales , Lactobacillaceae/enzimología , Lactobacillaceae/genética , Lactobacillales/metabolismo , Lipoproteínas , NADH Deshidrogenasa/metabolismo , Fosforilación
20.
J Am Chem Soc ; 144(2): 854-861, 2022 01 19.
Artículo en Inglés | MEDLINE | ID: mdl-34985894

RESUMEN

Rare-earth elements, which include the lanthanide series, are key components of many clean energy technologies, including wind turbines and photovoltaics. Because most of these 4f metals are at high risk of supply chain disruption, the development of new recovery technologies is necessary to avoid future shortages, which may impact renewable energy production. This paper reports the synthesis of a non-natural biogenic material as a potential platform for bioinspired lanthanide extraction. The biogenic material takes advantage of the atomically precise structure of a 2D crystalline protein lattice with the high lanthanide binding affinity of hydroxypyridinonate chelators. Luminescence titration data demonstrated that the engineered protein layers have affinities for all tested lanthanides in the micromolar-range (dissociation constants) and a higher binding affinity for the lanthanide ions with a smaller ionic radius. Furthermore, competitive titrations confirmed the higher selectivity (up to several orders of magnitude) of the biogenic material for lanthanides compared to other cations commonly found in f-element sources. Lastly, the functionalized protein layers could be reused in several cycles by desorbing the bound metal with citrate solutions. Taken together, these results highlight biogenic materials as promising bioadsorption platforms for the selective binding of lanthanides, with potential applications in the recovery of these critical elements from waste.


Asunto(s)
Quelantes/química , Metales de Tierras Raras/análisis , Proteínas/química , Concentración de Iones de Hidrógeno , Elementos de la Serie de los Lantanoides/análisis , Elementos de la Serie de los Lantanoides/aislamiento & purificación , Elementos de la Serie de los Lantanoides/metabolismo , Ligandos , Metales de Tierras Raras/aislamiento & purificación , Metales de Tierras Raras/metabolismo , Proteínas/metabolismo , Piridinas/química , Espectrofotometría
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