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1.
Cell ; 187(5): 1191-1205.e15, 2024 Feb 29.
Artigo em Inglês | MEDLINE | ID: mdl-38366592

RESUMO

Carbohydrate intolerance, commonly linked to the consumption of lactose, fructose, or sorbitol, affects up to 30% of the population in high-income countries. Although sorbitol intolerance is attributed to malabsorption, the underlying mechanism remains unresolved. Here, we show that a history of antibiotic exposure combined with high fat intake triggered long-lasting sorbitol intolerance in mice by reducing Clostridia abundance, which impaired microbial sorbitol catabolism. The restoration of sorbitol catabolism by inoculation with probiotic Escherichia coli protected mice against sorbitol intolerance but did not restore Clostridia abundance. Inoculation with the butyrate producer Anaerostipes caccae restored a normal Clostridia abundance, which protected mice against sorbitol-induced diarrhea even when the probiotic was cleared. Butyrate restored Clostridia abundance by stimulating epithelial peroxisome proliferator-activated receptor-gamma (PPAR-γ) signaling to restore epithelial hypoxia in the colon. Collectively, these mechanistic insights identify microbial sorbitol catabolism as a potential target for approaches for the diagnosis, treatment, and prevention of sorbitol intolerance.


Assuntos
Erros Inatos do Metabolismo dos Carboidratos , Microbioma Gastrointestinal , Sorbitol , Animais , Camundongos , Antibacterianos/farmacologia , Butiratos , Clostridium , Escherichia coli , Sorbitol/metabolismo
2.
Nature ; 507(7491): 201-6, 2014 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-24499818

RESUMO

Vaccines prevent infectious disease largely by inducing protective neutralizing antibodies against vulnerable epitopes. Several major pathogens have resisted traditional vaccine development, although vulnerable epitopes targeted by neutralizing antibodies have been identified for several such cases. Hence, new vaccine design methods to induce epitope-specific neutralizing antibodies are needed. Here we show, with a neutralization epitope from respiratory syncytial virus, that computational protein design can generate small, thermally and conformationally stable protein scaffolds that accurately mimic the viral epitope structure and induce potent neutralizing antibodies. These scaffolds represent promising leads for the research and development of a human respiratory syncytial virus vaccine needed to protect infants, young children and the elderly. More generally, the results provide proof of principle for epitope-focused and scaffold-based vaccine design, and encourage the evaluation and further development of these strategies for a variety of other vaccine targets, including antigenically highly variable pathogens such as human immunodeficiency virus and influenza.


Assuntos
Desenho de Fármacos , Epitopos/química , Epitopos/imunologia , Estabilidade Proteica , Vacinas contra Vírus Sincicial Respiratório/química , Vacinas contra Vírus Sincicial Respiratório/imunologia , Motivos de Aminoácidos , Animais , Anticorpos Monoclonais/análise , Anticorpos Monoclonais/imunologia , Anticorpos Neutralizantes/análise , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/análise , Anticorpos Antivirais/imunologia , Antígenos Virais/química , Antígenos Virais/imunologia , Cristalografia por Raios X , Ensaio de Imunoadsorção Enzimática , Macaca mulatta/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Modelos Moleculares , Testes de Neutralização , Conformação Proteica , Vírus Sinciciais Respiratórios/química , Vírus Sinciciais Respiratórios/imunologia
3.
Biologicals ; 53: 30-38, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29548791

RESUMO

According to manufacturers, inactivated poliovirus vaccines (IPVs) are freeze sensitive and require storage between 2°C and 8°C, whereas oral poliovirus vaccine requires storage at -20 °C. Introducing IPV into ongoing immunization services might result in accidental exposure to freezing temperatures and potential loss of vaccine potency. To better understand the effect of freezing IPVs, samples of single-dose vaccine vials from Statens Serum Institut (VeroPol) and multi-dose vaccine vials from Sanofi Pasteur (IPOL) were exposed to freezing temperatures mimicking what a vaccine vial might encounter in the field. D-antigen content was measured to determine the in vitro potency by ELISA. Immunogenicity testing was conducted for a subset of exposed IPVs using the rat model. Freezing VeroPol had no detectable effect on in vitro potency (D-antigen content) in all exposures tested. Freezing of the IPOL vaccine for 7 days at -20 °C showed statistically significant decreases in D-antigen content by ELISA in poliovirus type 1 (p < 0.0001) and type 3 (p = 0.048). Reduction of poliovirus type 2 potency also approached significance (p = 0.062). The observed loss in D-antigen content did not affect immunogenicity in the rat model. Further work is required to determine the significance of the loss observed and the implications for vaccine handling policies and practices.


Assuntos
Criopreservação , Congelamento , Imunogenicidade da Vacina , Vacina Antipólio de Vírus Inativado/imunologia , Animais , Feminino , Ratos , Ratos Wistar
4.
Sensors (Basel) ; 17(12)2017 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-29215582

RESUMO

This paper describes recent process modifications made to enhance the performance of interline and electron-multiplying charge-coupled-device (EMCCD) image sensors. By use of MeV ion implantation, quantum efficiency in the NIR region of the spectrum was increased by 2×, and image smear was reduced by 6 dB. By reducing the depth of the shallow photodiode (PD) implants, the photodiode-to-vertical-charge-coupled-device (VCCD) transfer gate voltage required for no-lag operation was reduced by 3 V, and the electronic shutter voltage was reduced by 9 V. The thinner, surface pinning layer also resulted in a reduction of smear by 4 dB in the blue portion of the visible spectrum. For EMCCDs, gain aging was eliminated by providing an oxide-only dielectric under its multiplication phase, while retaining the oxide-nitride-oxide (ONO) gate dielectrics elsewhere in the device.

5.
J Physiol ; 593(4): 843-56, 2015 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-25480802

RESUMO

KEY POINTS: Activation of NMDA receptors (NMDARs) is essential for encoding visual stimuli into signals for the brain, although their over-activation can cause cell death. The recruitment of NMDARs is important for encoding light intensity in retinal ganglion cells. D-serine binding is essential for proper activation of NMDARs, although its role in signal processing and the mechanisms that underlie its availability are not well understood. In these light-evoked experiments, the addition of exogenous D-serine had a large effect on low contrast and low intensity NMDAR responses that decreased as the intensity was increased. The degradation of endogenous D-serine decreased the responses more at higher intensities. The results provide compelling evidence favouring a new interpretation of NMDAR recruitment in which light-evoked D-serine release serves an important regulatory control over the recruitment of NMDARs. ABSTRACT: The present study aimed to investigate the functional properties of NMDA receptor coagonist release and to specifically evaluate whether light-evoked release mechanisms contribute to the availability of the coagonist D-serine. Two different methods were involved in our approach: (i) whole-cell recordings from identified retinal ganglion cells in the tiger salamander were used to study light adaptation with positive and negative contrast stimuli over a range of ± 1 log unit against a steady background illumination and (ii) the mechanisms for intensity encoding to a range of light intensities covering 6 log10 units were investigated. This latter study employed extracellular recordings of the proximal negative field potential, pharmacologically manipulated to generate a pure NMDA mediated response. For the adaptation study, we examined the light-evoked responses under control conditions, followed by light stimuli presented in the presence of D-serine, followed by light stimulation in the presence of dichlorokynurenic acid to block the coagonist site of NMDA receptors. For the brightness encoding studies, we examined the action of D-serine on each intensity used and then applied the enzyme D-serine deaminase to remove significant levels of D-serine. These studies provided new insights into the mechanisms that regulate coagonist availability in the vertebrate retina. Our results strongly support the idea that light-evoked coagonist release, a major component of which is D-serine, is needed to provide the full range of coagonist availability for optimal activation of NMDA receptors.


Assuntos
Receptores de N-Metil-D-Aspartato/fisiologia , Células Ganglionares da Retina/fisiologia , Serina/fisiologia , Ambystoma , Animais , Estimulação Luminosa
6.
PLoS Genet ; 7(9): e1002287, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21966277

RESUMO

It is an assumption of large, population-based datasets that samples are annotated accurately whether they correspond to known relationships or unrelated individuals. These annotations are key for a broad range of genetics applications. While many methods are available to assess relatedness that involve estimates of identity-by-descent (IBD) and/or identity-by-state (IBS) allele-sharing proportions, we developed a novel approach that estimates IBD0, 1, and 2 based on observed IBS within windows. When combined with genome-wide IBS information, it provides an intuitive and practical graphical approach with the capacity to analyze datasets with thousands of samples without prior information about relatedness between individuals or haplotypes. We applied the method to a commonly used Human Variation Panel consisting of 400 nominally unrelated individuals. Surprisingly, we identified identical, parent-child, and full-sibling relationships and reconstructed pedigrees. In two instances non-sibling pairs of individuals in these pedigrees had unexpected IBD2 levels, as well as multiple regions of homozygosity, implying inbreeding. This combined method allowed us to distinguish related individuals from those having atypical heterozygosity rates and determine which individuals were outliers with respect to their designated population. Additionally, it becomes increasingly difficult to identify distant relatedness using genome-wide IBS methods alone. However, our IBD method further identified distant relatedness between individuals within populations, supported by the presence of megabase-scale regions lacking IBS0 across individual chromosomes. We benchmarked our approach against the hidden Markov model of a leading software package (PLINK), showing improved calling of distantly related individuals, and we validated it using a known pedigree from a clinical study. The application of this approach could improve genome-wide association, linkage, heterozygosity, and other population genomics studies that rely on SNP genotype data.


Assuntos
Genoma Humano/genética , Estudo de Associação Genômica Ampla/métodos , Haplótipos/genética , Linhagem , Polimorfismo de Nucleotídeo Único , Algoritmos , Alelos , Simulação por Computador , Interpretação Estatística de Dados , Ligação Genética , Genótipo , Homozigoto , Humanos , Cadeias de Markov , Software
7.
J Am Pharm Assoc (2003) ; 54(3): 280-4, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24816355

RESUMO

OBJECTIVES: To describe patient interest and involvement in participating in a clinic-based community pharmacy drug take-back program to dispose of unused, unwanted, or expired (UUE) medications and to identify patients' reasons for participating in the program. METHODS: A convenience sample of patients at the University of Oklahoma Family Medicine Pharmacy was recruited to complete a needs assessment survey regarding interest in drug take-back programs and current practices for handling UUE medications. Participants who purchased a postage-paid drug disposal envelope were asked to complete a program survey identifying sources of UUE medications, experience with drug take-back programs, and reasons for participation. These participants were later contacted for a follow-up telephone survey regarding their experience with the program and medications sent back. RESULTS: 62 needs assessment surveys were collected. 61% of patients reported interest in a drug take-back program. 57% reported having no UUE medications at home. Commonly reported UUE handling practices included disposal in the garbage (53.2%) or sewer (29.0%) and home storage (17.7%). 15 disposal envelopes were sold to 10 participants whose most common reasons for participation included concern about the safety of household members, accidental or intentional ingestion, and environmental impact. For 4 patients who returned a median of 9.5 prescriptions, the most common class of returned drugs was antibiotics (19.0%). CONCLUSION: Interest in drug take-back programs exists, but awareness and availability of continuous programs is limited. Programs may be more successful if offered at no cost to patients. Future studies are needed on the types of medications sent back and specific reasons for accumulation.


Assuntos
Serviços Comunitários de Farmácia , Eliminação de Resíduos de Serviços de Saúde/métodos , Participação do Paciente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Farmacêuticos , Farmácia , Medicamentos sob Prescrição , Adulto Jovem
8.
Langmuir ; 29(21): 6495-501, 2013 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-23617458

RESUMO

Environmentally responsive hydrogels hold multiple important applications. However, the functionality of these materials alone is often limited in comparison to other materials like silicon; thus, there is a need to integrate soft and hard materials for the advancement of environmentally sensitive materials. Here we demonstrate the capability of integrating a thermoresponsive hydrogel, poly(N-isopropylacrylamide), with thin film silicon ribbons, enabling the stiff silicon ribbons to become adaptive and drivable by the soft environmentally sensitive substrate. This integration provides a means of mechanical buckling of the thin silicon film due to changes in environmental stimuli (e.g., temperature, pH). We also investigate how advanced lithographic techniques can be used to generate patterned deformation on the aforementioned integrated structures. Furthermore, we explore multilayer hybrid hydrogel structures formed by the integration of different types of hydrogels that have tunable curvatures under the influence of different stimuli. Silicon thin film integration on such tunable curvature substrates reveal characteristic reversible buckling of the thin film in the presence of multiple stimuli. These results open new opportunities for developing stretchable and intelligent devices for multiple applications.


Assuntos
Acrilamidas/química , Hidrogéis/química , Polímeros/química , Silício/química , Acrilamidas/síntese química , Resinas Acrílicas , Hidrogéis/síntese química , Tamanho da Partícula , Polímeros/síntese química , Propriedades de Superfície
9.
PLoS Genet ; 6(6): e1000991, 2010 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-20577567

RESUMO

Although more than 2,400 genes have been shown to contain variants that cause Mendelian disease, there are still several thousand such diseases yet to be molecularly defined. The ability of new whole-genome sequencing technologies to rapidly indentify most of the genetic variants in any given genome opens an exciting opportunity to identify these disease genes. Here we sequenced the whole genome of a single patient with the dominant Mendelian disease, metachondromatosis (OMIM 156250), and used partial linkage data from her small family to focus our search for the responsible variant. In the proband, we identified an 11 bp deletion in exon four of PTPN11, which alters frame, results in premature translation termination, and co-segregates with the phenotype. In a second metachondromatosis family, we confirmed our result by identifying a nonsense mutation in exon 4 of PTPN11 that also co-segregates with the phenotype. Sequencing PTPN11 exon 4 in 469 controls showed no such protein truncating variants, supporting the pathogenicity of these two mutations. This combination of a new technology and a classical genetic approach provides a powerful strategy to discover the genes responsible for unexplained Mendelian disorders.


Assuntos
Ligação Genética , Predisposição Genética para Doença , Genoma Humano , Proteína Tirosina Fosfatase não Receptora Tipo 11/genética , Éxons , Feminino , Estudo de Associação Genômica Ampla , Humanos , Masculino , Mutação , Linhagem , Análise de Sequência de DNA
10.
FEMS Microbiol Rev ; 47(3)2023 05 19.
Artigo em Inglês | MEDLINE | ID: mdl-37188642

RESUMO

Extracellular electron transfer (EET) is a bioelectrochemical process performed by electrochemically active bacteria (EAB) found in host-associated environments, including plant and animal ecosystems and fermenting plant- and animal-derived foods. Through direct or mediated electron transfer pathways, certain bacteria use EET to enhance ecological fitness with host-impacting effects. In the plant rhizosphere, electron acceptors support the growth of EAB such as Geobacter, cable bacteria, and some clostridia that can result changing iron and heavy metal uptake by plants. In animal microbiomes, EET is associated with diet-derived iron in the intestines of soil-dwelling termites, earthworms, and beetle larvae. EET is also associated with the colonization and metabolism of some bacteria in human and animal microbiomes, such as Streptococcus mutans in the mouth, Enterococcus faecalis and Listeria monocytogenes in the intestine, and Pseudomonas aeruginosa in the lungs. During the fermentation of plant tissues and bovine milk, lactic acid bacteria like Lactiplantibacillus plantarum and Lactococcus lactis may use EET to increase their growth and food acidification, as well as decrease environmental oxidation-reduction potential. Thus, EET is likely an important metabolic pathway for host-associated bacteria and has implications for ecosystem function, health and disease, and biotechnological applications.


Assuntos
Ecossistema , Elétrons , Animais , Humanos , Transporte de Elétrons , Bactérias/metabolismo , Ferro/metabolismo
11.
PLoS One ; 18(10): e0292585, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37824485

RESUMO

Lactobacilli and Acetobacter sp. are commercially important bacteria that often form communities in natural fermentations, including food preparations, spoilage, and in the digestive tract of the fruit fly Drosophila melanogaster. Communities of these bacteria are widespread and prolific, despite numerous strain-specific auxotrophies, suggesting they have evolved nutrient interdependencies that regulate their growth. The use of a chemically-defined medium (CDM) supporting the growth of both groups of bacteria would facilitate the identification of the molecular mechanisms for the metabolic interactions between them. While numerous CDMs have been developed that support specific strains of lactobacilli or Acetobacter, there has not been a medium formulated to support both genera. We developed such a medium, based on a previous CDM designed for growth of lactobacilli, by modifying the nutrient abundances to improve growth yield. We further simplified the medium by substituting casamino acids in place of individual amino acids and the standard Wolfe's vitamins and mineral stocks in place of individual vitamins and minerals, resulting in a reduction from 40 to 8 stock solutions. These stock solutions can be used to prepare several CDM formulations that support robust growth of numerous lactobacilli and Acetobacters. Here, we provide the composition and several examples of its use, which is important for tractability in dissecting the genetic and metabolic basis of natural bacterial species interactions.


Assuntos
Acetobacter , Animais , Acetobacter/genética , Lactobacillus/fisiologia , Drosophila melanogaster , Bactérias , Vitaminas/metabolismo
12.
mBio ; : e0223423, 2023 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-37982640

RESUMO

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.

13.
bioRxiv ; 2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36711674

RESUMO

Most cancers exhibit aneuploidy, but its functional significance in tumor development is controversial. Here, we describe ReDACT (Restoring Disomy in Aneuploid cells using CRISPR Targeting), a set of chromosome engineering tools that allow us to eliminate specific aneuploidies from cancer genomes. Using ReDACT, we created a panel of isogenic cells that have or lack common aneuploidies, and we demonstrate that trisomy of chromosome 1q is required for malignant growth in cancers harboring this alteration. Mechanistically, gaining chromosome 1q increases the expression of MDM4 and suppresses TP53 signaling, and we show that TP53 mutations are mutually-exclusive with 1q aneuploidy in human cancers. Thus, specific aneuploidies play essential roles in tumorigenesis, raising the possibility that targeting these "aneuploidy addictions" could represent a novel approach for cancer treatment.

14.
Science ; 381(6660): eadg4521, 2023 08 25.
Artigo em Inglês | MEDLINE | ID: mdl-37410869

RESUMO

Most cancers exhibit aneuploidy, but its functional significance in tumor development is controversial. Here, we describe ReDACT (Restoring Disomy in Aneuploid cells using CRISPR Targeting), a set of chromosome engineering tools that allow us to eliminate specific aneuploidies from cancer genomes. Using ReDACT, we created a panel of isogenic cells that have or lack common aneuploidies, and we demonstrate that trisomy of chromosome 1q is required for malignant growth in cancers harboring this alteration. Mechanistically, gaining chromosome 1q increases the expression of MDM4 and suppresses p53 signaling, and we show that TP53 mutations are mutually exclusive with 1q aneuploidy in human cancers. Thus, tumor cells can be dependent on specific aneuploidies, raising the possibility that these "aneuploidy addictions" could be targeted as a therapeutic strategy.


Assuntos
Proteínas de Ciclo Celular , Edição de Genes , Neoplasias , Oncogenes , Trissomia , Proteína Supressora de Tumor p53 , Humanos , Proteínas de Ciclo Celular/genética , Mutação , Neoplasias/genética , Neoplasias/terapia , Proteínas Proto-Oncogênicas/metabolismo , Edição de Genes/métodos , Proteína Supressora de Tumor p53/genética , Carcinogênese/genética
15.
Hum Mutat ; 33(7): 1075-86, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22374857

RESUMO

Tens of thousands of lymphoblastoid cell lines (LCLs) have been established by the research community, providing nearly unlimited source material from samples of interest. LCLs are used to address questions in population genomics, mechanisms of disease, and pharmacogenomics. Thus, it is of fundamental importance to define the extent of chromosomal variation in LCLs. We measured variation in genotype and copy number in multiple LCLs derived from peripheral blood mononuclear cells (PBMCs) of single individuals as well as two comparison groups: (1) three types of differentiated cell lines (DCLs) and (2) triplicate HapMap samples. We then validated and extended our findings using data from a large study consisting of samples from blood or LCLs. We observed high concordances between genotypes and copy number estimates within all sample groups. While the genotypes of LCLs tended to faithfully reflect the genotypes of PBMCs, 13.7% (4 of 29) of immortalized cell lines harbored mosaic regions greater than 20 megabases, which were not present in PBMCs, DCLs, or HapMap replicate samples. We created a list of putative LCL-specific changes (affecting regions such as immunoglobulin loci) that is available as a community resource.


Assuntos
Variações do Número de Cópias de DNA/genética , Linhagem Celular , Células Cultivadas , Genótipo , Humanos , Polimorfismo de Nucleotídeo Único/genética
16.
Elife ; 112022 02 11.
Artigo em Inglês | MEDLINE | ID: mdl-35147079

RESUMO

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.


Assuntos
Transporte de Elétrons/fisiologia , Fermentação , Lactobacillaceae/metabolismo , Albinismo Oculocutâneo , Biomassa , Brassica/química , Sucos de Frutas e Vegetais , Lactobacillaceae/enzimologia , Lactobacillaceae/genética , Lactobacillales/metabolismo , Lipoproteínas , NADH Desidrogenase/metabolismo , Fosforilação
17.
J Food Prot ; 85(5): 755-772, 2022 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-35259246

RESUMO

ABSTRACT: This multiagency report developed by the Interagency Collaboration for Genomics for Food and Feed Safety provides an overview of the use of and transition to whole genome sequencing (WGS) technology for detection and characterization of pathogens transmitted commonly by food and for identification of their sources. We describe foodborne pathogen analysis, investigation, and harmonization efforts among the following federal agencies: National Institutes of Health; Department of Health and Human Services, Centers for Disease Control and Prevention (CDC) and U.S. Food and Drug Administration (FDA); and the U.S. Department of Agriculture, Food Safety and Inspection Service, Agricultural Research Service, and Animal and Plant Health Inspection Service. We describe single nucleotide polymorphism, core-genome, and whole genome multilocus sequence typing data analysis methods as used in the PulseNet (CDC) and GenomeTrakr (FDA) networks, underscoring the complementary nature of the results for linking genetically related foodborne pathogens during outbreak investigations while allowing flexibility to meet the specific needs of Interagency Collaboration partners. We highlight how we apply WGS to pathogen characterization (virulence and antimicrobial resistance profiles) and source attribution efforts and increase transparency by making the sequences and other data publicly available through the National Center for Biotechnology Information. We also highlight the impact of current trends in the use of culture-independent diagnostic tests for human diagnostic testing on analytical approaches related to food safety and what is next for the use of WGS in the area of food safety.


Assuntos
Doenças Transmitidas por Alimentos , Animais , Surtos de Doenças/prevenção & controle , Inocuidade dos Alimentos , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/prevenção & controle , Genômica , Estados Unidos , Sequenciamento Completo do Genoma
18.
Ophthalmic Plast Reconstr Surg ; 27(6): 414-6, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21785381

RESUMO

PURPOSE: To examine the change in orbital apical temperature over time, with a local ice/water admixture compress placed over the eyelid. METHODS: A six-month-old, 220-pound Landrace/Hampshire/Duroc mixed breed pig was intubated and maintained under anesthesia with a steady average body temperature. An incision in the lateral third of the right lower eyelid along the inferior orbital rim was made, and a digital thermometer was inserted and guided toward the orbital apex by fluoroscopic imaging of the orbit. After the baseline apical temperature was measured, an ice/water admixture compress was placed on the right eyelid, and serial temperatures were taken every 5 minutes for 20 minutes. A final measurement was taken at 45 minutes after the ice/water admixture compress placement. RESULTS: At an average core body temperature of 38.9°C, the baseline apical temperature was 37.2°C. After placement of a 0°C ice/water admixture, there was an exponential decrease in apical temperature, reaching a plateau after a period of 20 minutes, with a decrease of 1.4°C from baseline. The same apical temperature noted 20 minutes after placement of the cool compress was measured 45 minutes after ice/water compress placement. CONCLUSIONS: There was only a limited decrease in apical temperature by placing a local 0°C ice/water admixture over the eyelid. This information is useful in determining whether local hypothermia may be a potential alternative for traumatic optic neuropathy.


Assuntos
Temperatura Corporal/fisiologia , Hipotermia Induzida , Órbita/fisiologia , Animais , Bandagens , Pálpebras , Suínos
19.
mSystems ; 6(1)2021 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-33563789

RESUMO

Validated methods are needed to detect spoilage microbes present in low numbers in foods and ingredients prior to defect onset. We applied propidium monoazide combined with 16S rRNA gene sequencing, qPCR, isolate identification, and pilot-scale cheese making to identify the microorganisms that cause slit defects in industrially produced Cheddar cheese. To investigate milk as the source of spoilage microbes, bacterial composition in milk was measured immediately before and after high-temperature, short-time (HTST) pasteurization over 10-h periods on 10 days and in the resulting cheese blocks. Besides HTST pasteurization-induced changes to milk microbiota composition, a significant increase in numbers of viable bacteria was observed over the 10-h run times of the pasteurizer, including 68-fold-higher numbers of the genus Thermus However, Thermus was not associated with slit development. Milk used to make cheese which developed slits instead contained a lower number of total bacteria, higher alpha diversity, and higher proportions of Lactobacillus, Bacillus, Brevibacillus, and Clostridium Only Lactobacillus proportions were significantly increased during cheese aging, and Limosilactobacillus (Lactobacillus) fermentum, in particular, was enriched in slit-containing cheeses and the pre- and post-HTST-pasteurization milk used to make them. Pilot-scale cheeses developed slits when inoculated with strains of L. fermentum, other heterofermentative lactic acid bacteria, or uncultured bacterial consortia from slit-associated pasteurized milk, thereby confirming that low-abundance taxa in milk can negatively affect cheese quality. The likelihood that certain microorganisms in milk cause slit defects can be predicted based on comparisons of the bacteria present in the milk used for cheese manufacture.IMPORTANCE Food production involves numerous control points for microorganisms to ensure quality and safety. These control points (e.g., pasteurization) are difficult to develop for fermented foods wherein some microbial contaminants are also expected to provide positive contributions to the final product and spoilage microbes may constitute only a small proportion of all microorganisms present. We showed that microbial composition assessments with 16S rRNA marker gene DNA sequencing are sufficiently robust to detect very-low-abundance bacterial taxa responsible for a major but sporadic Cheddar cheese spoilage defect. Bacterial composition in the (pasteurized) milk and cheese was associated with slit defect development. The application of Koch's postulates showed that individual bacterial isolates as well as uncultured bacterial consortia were sufficient to cause slits, even when present in very low numbers. This approach may be useful for detection and control of low-abundance spoilage microorganisms present in other foods.

20.
Pathogens ; 10(11)2021 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-34832547

RESUMO

Water is vital to agriculture. It is essential that the water used for the production of fresh produce commodities be safe. Microbial pathogens are able to survive for extended periods of time in water. It is critical to understand their biology and ecology in this ecosystem in order to develop better mitigation strategies for farmers who grow these food crops. In this review the prevalence, persistence and ecology of four major foodborne pathogens, Shiga toxin-producing Escherichia coli (STEC), Salmonella, Campylobacter and closely related Arcobacter, and Listeria monocytogenes, in water are discussed. These pathogens have been linked to fresh produce outbreaks, some with devastating consequences, where, in a few cases, the contamination event has been traced to water used for crop production or post-harvest activities. In addition, antimicrobial resistance, methods improvements, including the role of genomics in aiding in the understanding of these pathogens, are discussed. Finally, global initiatives to improve our knowledge base of these pathogens around the world are touched upon.

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