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1.
Pediatr Blood Cancer ; : e30431, 2023 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-37277314

RESUMO

INTRODUCTION: Brainstem tumors comprise 10.9% of all brain tumors, and pediatric diffuse intrinsic pontine gliomas (DIPG) have a fatal prognosis. Some countries have developed national and international register databases to characterize their populations to aid clinical and public policy decisions. This study provides information regarding the clinical characteristics of a retrospective cohort of children with DIPG in México from 2001 to 2021, and assesses the proposed prognostic factors previously described for survival outcome. METHODS: Health institutions from Mexico were invited to contribute to a retrospective electronic registry of patients with DIPG based on the International DIPG Registry. Fisher's exact test was used to compare long- and short-term survivors. Overall survival was estimated using the Kaplan-Meier method. Differences between survival curves were evaluated using the log-rank test and Cox proportional hazard regression analysis. RESULTS: Total 110 patients were included. The median age of the patients at diagnosis was 7 years. Sixty patients (54.5%) presented with symptoms in less than 6 months; the most frequent symptom was ataxia (56.4%). Ninety patients received treatment (81.8%), the overall survival at 4 years was 11.4%, and 16 patients (14.5%) were admitted for palliative end-of-life care. We found no significant survival differences for any of the prognostic factors. CONCLUSION: This study highlights the need to develop strategies to standardize healthcare processes and enhance the quality of care to improve clinical diagnosis in Mexico. We also observed a barrier to the acceptance of palliative end-of-life care in the family and medical teams.

2.
Salud Publica Mex ; 63(2, Mar-Abr): 160-162, 2021 Feb 27.
Artigo em Espanhol | MEDLINE | ID: mdl-33989476

RESUMO

OBJECTIVE: To describe a Covid-19 outbreak in a gerontological center in Mexico City. MATERIAL AND METHODS: Cross-sectional study in older adults. The association of risk factors for dying from Covid-19 was analyzed using a multiple logistic regression model. RESULTS: One hundred and two elders with an average age of 82.5 ± 8.8 years were included. Fifty-five (54%) tested positive and 47 (46%) were negative for the new coronavirus. Using the multiple logistic regression model, people with frailty had an OR of 11.6 of dying from Covid-19 compared to robust people (p-value = 0.024). CONCLUSION: The Covid-19 outbreak was initially caused by a resident of the center and spread by cross infection. In vulnerable populations, early detection, isolation, and follow-up of contacts should be carried out, as well as the identification of risk factors in order to reduce the spread and mortality caused by SARSCoV-2.


Assuntos
COVID-19/epidemiologia , Surtos de Doenças , Instituição de Longa Permanência para Idosos , Idoso , Idoso de 80 Anos ou mais , Estudos Transversais , Feminino , Humanos , Masculino , México
3.
Mol Plant Microbe Interact ; 30(7): 557-565, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28548604

RESUMO

Quorum-sensing systems play important roles in host colonization and host establishment of Burkholderiales species. Beneficial Paraburkholderia species share a conserved quorum-sensing (QS) system, designated BraI/R, that controls different phenotypes. In this context, the plant growth-promoting bacterium Paraburkholderia phytofirmans PsJN possesses two different homoserine lactone QS systems BpI.1/R.1 and BpI.2/R.2 (BraI/R-like QS system). The BpI.1/R.1 QS system was previously reported to be important to colonize and produce beneficial effects in Arabidopsis thaliana plants. Here, we analyzed the temporal variations of the QS gene transcript levels in the wild-type strain colonizing plant roots. The gene expression patterns showed relevant differences in both QS systems compared with the wild-type strain in the unplanted control treatment. The gene expression data were used to reconstruct a regulatory network model of QS systems in P. phytofirmans PsJN, using a Boolean network model. Also, we examined the phenotypic traits and transcript levels of genes involved in QS systems, using P. phytofirmans mutants in homoserine lactone synthases genes. We observed that the BpI.1/R.1 QS system regulates biofilm formation production in strain PsJN and this phenotype was associated with the lower expression of a specific extracytoplasmic function sigma factor ecf26.1 gene (implicated in biofilm formation) in the bpI.1 mutant strain.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Biofilmes , Burkholderia/genética , Raízes de Plantas/crescimento & desenvolvimento , Percepção de Quorum/genética , 4-Butirolactona/análogos & derivados , 4-Butirolactona/metabolismo , Arabidopsis/microbiologia , Burkholderia/metabolismo , Burkholderia/fisiologia , Perfilação da Expressão Gênica/métodos , Regulação Bacteriana da Expressão Gênica , Redes Reguladoras de Genes , Interações Hospedeiro-Patógeno , Modelos Genéticos , Mutação , Raízes de Plantas/microbiologia
4.
Appl Environ Microbiol ; 83(1)2017 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-27795307

RESUMO

Several bacteria use the plant hormone indole-3-acetic acid (IAA) as a sole carbon and energy source. A cluster of genes (named iac) encoding IAA degradation has been reported in Pseudomonas putida 1290, but the functions of these genes are not completely understood. The plant-growth-promoting rhizobacterium Paraburkholderia phytofirmans PsJN harbors iac gene homologues in its genome, but with a different gene organization and context than those of P. putida 1290. The iac gene functions enable P. phytofirmans to use IAA as a sole carbon and energy source. Employing a heterologous expression system approach, P. phytofirmans iac genes with previously undescribed functions were associated with specific biochemical steps. In addition, two uncharacterized genes, previously unreported in P. putida and found to be related to major facilitator and tautomerase superfamilies, are involved in removal of an IAA metabolite called dioxindole-3-acetate. Similar to the case in strain 1290, IAA degradation proceeds through catechol as intermediate, which is subsequently degraded by ortho-ring cleavage. A putative two-component regulatory system and a LysR-type regulator, which apparently respond to IAA and dioxindole-3-acetate, respectively, are involved in iac gene regulation in P. phytofirmans These results provide new insights about unknown gene functions and complex regulatory mechanisms in IAA bacterial catabolism. IMPORTANCE: This study describes indole-3-acetic acid (auxin phytohormone) degradation in the well-known betaproteobacterium P. phytofirmans PsJN and comprises a complete description of genes, some of them with previously unreported functions, and the general basis of their gene regulation. This work contributes to the understanding of how beneficial bacteria interact with plants, helping them to grow and/or to resist environmental stresses, through a complex set of molecular signals, in this case through degradation of a highly relevant plant hormone.


Assuntos
Proteínas de Bactérias/genética , Burkholderiaceae/genética , Burkholderiaceae/metabolismo , Regulação Bacteriana da Expressão Gênica , Ácidos Indolacéticos/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Bactérias/metabolismo , Burkholderiaceae/fisiologia , Genes Bacterianos , Desenvolvimento Vegetal , Reguladores de Crescimento de Plantas/química , Pseudomonas putida/genética , Pseudomonas putida/metabolismo , Estresse Fisiológico/genética
5.
Front Oncol ; 14: 1304605, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38444670

RESUMO

Introduction: The mechanistic/mammalian target of rapamycin (mTOR) is a serine/threonine kinase, which is downregulated or upregulated and is implicated in different types of cancer including hematologic neoplasms, skin prostate, and head and neck cancer. Aim: The aim of this study was to explore the current knowledge of mTOR signaling in acute lymphoblastic leukemia and Hodgkin lymphoma. Methods: A systematic review was performed according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, searching PubMed, Discovery Service for National Autonomous University of Mexico, Registro Nacional de Instituciones y Empresas Científicas y Tecnológicas (RENIECYT), and Scientific Electronic Library Online (SciELO) from 1994 to 2023. A total of 269 papers were identified for acute lymphoblastic leukemia, but based on specific criteria, 15 were included; for Hodgkin lymphoma, 110 papers were identified, but 5 were included after manual searching. Results: A total of 20 papers were evaluated, where mTOR activity is increased in patients with Hodgkin lymphoma and acute lymphoblastic leukemia by different molecular mechanisms. Conclusions: mTOR activity is increased in patients with both hematologic neoplasms and NOTCH; interleukin 4, 7, and 9, and nuclear proteins have been studied for their role in the activation of mTOR signaling.

6.
Iran J Allergy Asthma Immunol ; 23(1): 122-126, 2024 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-38485907

RESUMO

Actinrelated protein 2/3 complex subunit 1B (ARPC1B) deficiency is an inborn error of immunity (IEI) characterized by a combination of immunodeficiency and immune dysregulation and classified as an IEI with allergic manifestations. Here, we describe two patients with pathogenic variants in the ARPC1B gene. The first patient presented with eczema and bronchospasm at six months of age. The second patient presented with eczema and milk protein allergy at five months of age. The c.899_944 (p.Glu300Glyfs*7) pathogenic variant was previously described, whereas the c.863del (p.Pro288Leufs*9) variant was novel. ARPC1B deficiency should be considered because of the severe allergic manifestations at an early age.


Assuntos
Eczema , Hipersensibilidade Alimentar , Síndromes de Imunodeficiência , Hipersensibilidade a Leite , Animais , Humanos , Lactente , Complexo 2-3 de Proteínas Relacionadas à Actina/genética , Alérgenos , Eczema/genética , Síndromes de Imunodeficiência/genética , Leite , Hipersensibilidade a Leite/diagnóstico , Hipersensibilidade a Leite/genética
7.
CNS Neurol Disord Drug Targets ; 23(4): 420-430, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-37038673

RESUMO

BACKGROUND: B7H3 is a co-stimulatory molecule for immune reactions found on the surface of tumor cells in a wide variety of tumors. Preclinical and clinical studies have reported it as a tumor target towards which various immunotherapy modalities could be directed. So far, good results have been obtained in hematological neoplasms; however, a contrasting situation is evident in solid tumors, including those of the CNS, which show high refractoriness to current treatments. The appearance of cellular immunotherapies has transformed oncology due to the reinforcement of the immune response that is compromised in people with cancer. OBJECTIVE: This article aims to review the literature to describe the advancement in knowledge on B7H3 as a target of CAR-T cells in pediatric gliomas to consider them as an alternative in the treatment of these patients. RESULTS: Although B7H3 is considered a suitable candidate as a target agent for various immunotherapy techniques, there are still limitations in using CAR-T cells to achieve the desired success. CONCLUSION: Results obtained with CAR-T cells can be further improved by the suggested proposals; therefore, more clinical trials are needed to study this new therapy in children with gliomas.


Assuntos
Glioma , Imunoterapia , Humanos , Criança , Glioma/terapia , Linfócitos T
8.
Mol Plant Microbe Interact ; 26(5): 546-53, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23301615

RESUMO

Although not fully understood, molecular communication in the rhizosphere plays an important role regulating traits involved in plant-bacteria association. Burkholderia phytofirmans PsJN is a well-known plant-growth-promoting bacterium, which establishes rhizospheric and endophytic colonization in different plants. A competent colonization is essential for plant-growth-promoting effects produced by bacteria. Using appropriate mutant strains of B. phytofirmans, we obtained evidence for the importance of N-acyl homoserine lactone-mediated (quorum sensing) cell-to-cell communication in efficient colonization of Arabidopsis thaliana plants and the establishment of a beneficial interaction. We also observed that bacterial degradation of the auxin indole-3-acetic acid (IAA) plays a key role in plant-growth-promoting traits and is necessary for efficient rhizosphere colonization. Wildtype B. phytofirmans but not the iacC mutant in IAA mineralization is able to restore promotion effects in roots of A. thaliana in the presence of exogenously added IAA, indicating the importance of this trait for promoting primary root length. Using a transgenic A. thaliana line with suppressed auxin signaling (miR393) and analyzing the expression of auxin receptors in wild-type inoculated plants, we provide evidence that auxin signaling in plants is necessary for the growth promotion effects produced by B. phytofirmans. The interplay between ethylene and auxin signaling was also confirmed by the response of the plant to a 1-aminocyclopropane-1-carboxylate deaminase bacterial mutant strain.


Assuntos
Arabidopsis/metabolismo , Arabidopsis/microbiologia , Burkholderia/fisiologia , Ácidos Indolacéticos/metabolismo , Percepção de Quorum , Arabidopsis/crescimento & desenvolvimento , Transdução de Sinais
9.
Methods Mol Biol ; 2553: 155-171, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36227543

RESUMO

Synthetic biology aims at engineering new biological systems and functions that can be used to provide new technological solutions to worldwide challenges. Detection and processing of multiple signals are crucial for many synthetic biology applications. A variety of logic circuits operating in living cells have been implemented. One particular class of logic circuits uses site-specific recombinases mediating specific DNA inversion or excision. Recombinase logic offers many interesting features, including single-layer architectures, memory, low metabolic footprint, and portability in many species. Here, we present two automated design strategies for both Boolean and history-dependent recombinase-based logic circuits. One approach is based on the distribution of computation within multicellular consortia, and the other is a single-cell design. Both are complementary and adapted for non-expert users via a web design interface, called CALIN and RECOMBINATOR, for multicellular and single-cell design strategies, respectively. In this book chapter, we are guiding the reader step by step through recombinase logic circuit design, from selecting the design strategy fitting to their final system of interest to obtaining the final design using one of our design web interfaces.


Assuntos
Lógica , Recombinases , DNA , Recombinases/genética , Recombinases/metabolismo , Biologia Sintética/métodos
10.
Disabil Rehabil Assist Technol ; 18(3): 274-284, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-33156714

RESUMO

BACKGROUND: Social participation is an important aspect of health and well-being across the lifespan, but older adults might encounter some barriers, which has been highlighted in the current Covid-19 pandemic situation, where technology has become the primary way to maintain contact with family and friends. In fact, technology can serve both as a facilitator and barrier to social participation in later life, and this issue needs to be further understood. AIM: To identify the barriers and facilitators encountered by older adults in using technology to promote social participation. METHODS: A systematic review was conducted. Studies were included if they were peer-reviewed, written in English or French, included participants 50 years or older, included technology to promote social participation, and reported potential barriers or facilitators regarding such technologies. Four databases were included: MEDLINE, CINAHL, PsychINFO and, ERIC. Each study was reviewed by two independent reviewers. The quality of the study was appraised using the Crowe Critical Appraisal Tool. RESULTS: Seventeen studies were included in this report. Four main themes emerged from the data: perceived benefits of the technology, self-confidence and knowledge about using the technology efficiently and safely, affordability of the technology, and ability of the technology to adapt to the physical and cognitive declines in later life. CONCLUSION: These findings can help health care professionals to make better decisions when deciding to recommend technology for their older clients.IMPLICATIONS FOR REHABILITATIONAcceptance of technology to promote social participation in later life is a multi-complex process. There is no "one size fits all" approach, a person-centered intervention must be used.When introducing new technologies, using an adapted/tailored training approach could potentially increase self-efficacy in using technology.Rehabilitation professionals' misconceptions concerning the use of technology in later life can be a barrier to acceptance. It's important to be aware of our own believes and attitudes in this context.


Assuntos
COVID-19 , Pandemias , Humanos , Idoso , Participação Social , Pessoal de Saúde/psicologia
11.
PLoS One ; 18(2): e0280935, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36800374

RESUMO

Engineered bacteria are promising candidates for in situ detection and treatment of diseases. The female uro-genital tract presents several pathologies, such as sexually transmitted diseases or genital cancer, that could benefit from such technology. While bacteria from the gut microbiome are increasingly engineered, the use of chassis isolated from the female uro-genital resident flora has been limited. A major hurdle to implement the experimental throughput required for efficient engineering in these non-model bacteria is their low transformability. Here we report an optimized electrotransformation protocol for Lactobacillus jensenii, one the most widespread species across vaginal microflora. Starting from classical conditions, we optimized buffers, electric field parameters, cuvette type and DNA quantity to achieve an 80-fold improvement in transformation efficiency, with up to 3.5·103 CFUs/µg of DNA in L. jensenii ATCC 25258. We also identify several plasmids that are maintained and support reporter gene expression in L. jensenii. Finally, we demonstrate that our protocol provides increased transformability in three independent clinical isolates of L. jensenii. This work will facilitate the genetic engineering of L. jensenii and enable its use for addressing challenges in gynecological healthcare.


Assuntos
Lactobacillus , Vagina , Feminino , Humanos , Vagina/microbiologia , Bactérias/genética , Plasmídeos/genética
12.
Cureus ; 15(9): e45473, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37727841

RESUMO

Background The escalating overload and saturation of emergency services, primarily caused by non-urgent cases overwhelming the system, have spurred a critical necessity for innovative solutions that can effectively differentiate genuine emergencies from situations that could be managed through alternative means, such as using AI chatbots. This study aims to evaluate and compare the accuracy in differentiating between a medical emergency and a non-emergency of three of the most popular AI chatbots at the moment. Methods In this study, patient questions from the online forum r/AskDocs on Reddit were collected to determine whether their clinical cases were emergencies. A total of 176 questions were reviewed by the authors, with 75 deemed emergencies and 101 non-emergencies. These questions were then posed to AI chatbots, including ChatGPT, Google Bard, and Microsoft Bing AI, with their responses evaluated against each other and the authors' responses. A criteria-based system categorized the AI chatbot answers as "yes," "no," or "cannot determine." The performance of each AI chatbot was compared in both emergency and non-emergency cases, and statistical analysis was conducted to assess the significance of differences in their performance. Results In general, AI chatbots considered around 12-15% more cases to be an emergency than reviewers, while they considered a very low number of cases as non-emergency compared to reviewers (around 35% fewer cases). Google Bard detected the most true emergency cases (87%) and true non-emergency cases (36%). However, no real difference in performance between the three AI chatbots was found in detecting true emergencies (p-value = 0.35) and non-emergency cases (p-value = 0.16).  Conclusions These AI systems require further refinement to identify emergency situations accurately, but they could potentially be an innovative tool for emergency care and improving patient outcomes. The integration of AI chatbots like ChatGPT, Google Bard, and Microsoft Bing Chat offers a promising avenue to mitigate ED strain and enhance emergency management.

13.
Clin Interv Aging ; 18: 273-282, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36851976

RESUMO

Purpose: To measure the effects of an exercise program on the physical capacities of older adults such as strength, flexibility, balance, and aerobic capacity. Patients and Methods: This was a quasi-experimental study on a population of 5550 older adults and a sample of 4830 participants in an active aging program designed by the Municipal Health Secretary. The exercise program lasted 12 months, and pre-and post-program intervention measures were recorded using the senior fitness test. Results: Most participants were women (92.4%) and their mean age was 70.7 years (standard deviation, 7.3 years; range, 60-97 years). All areas showed significant differences before and after the program in terms of the participants' physical capabilities (p < 0.05), muscular strength and flexibility had a more significant mean difference and a large effect (>0.80), except for aerobic capacity, which had a small effect. Conclusion: The present study revealed that a supervised physical exercise program at the community level has positive effects on the physical capacities of coordination, balance, flexibility, strength, and aerobic capacity, which are essential components for a better functional capacity at this stage of life, with improvements that encompassed the improved self-perception of their health status, a reduction of overweight and obesity. The reinforcement of these programs is recommended, consequently, promoting pre-sport games and sports championships among the elderly population, as a public health strategy.


Assuntos
Exercício Físico , Força Muscular , Humanos , Idoso , Feminino , Masculino , Envelhecimento , Terapia Comportamental , Terapia por Exercício
14.
Antonie Van Leeuwenhoek ; 101(4): 713-23, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22186997

RESUMO

Plant rhizosphere and internal tissues may constitute a relevant habitat for soil bacteria displaying high catabolic versatility towards xenobiotic aromatic compounds. Root exudates contain various molecules that are structurally related to aromatic xenobiotics and have been shown to stimulate bacterial degradation of aromatic pollutants in the rhizosphere. The ability to degrade specific aromatic components of root exudates could thus provide versatile catabolic bacteria with an advantage for rhizosphere colonization and growth. In this work, Cupriavidus pinatubonensis JMP134, a well-known aromatic compound degrader (including the herbicide 2,4-dichlorophenoxyacetate, 2,4-D), was shown to stably colonize Arabidopsis thaliana and Acacia caven plants both at the rhizoplane and endorhizosphere levels and to use root exudates as a sole carbon and energy source. No deleterious effects were detected on these colonized plants. When a toxic concentration of 2,4-D was applied to colonized A. caven, a marked resistance was induced in the plant, showing that strain JMP134 was both metabolically active and potentially beneficial to its host. The role for the ß-ketoadipate aromatic degradation pathway during plant root colonization by C. pinatubonensis JMP134 was investigated by gene inactivation. A C. pinatubonensis mutant derivative strain displayed a reduced ability to catabolise root exudates isolated from either plant host. In this mutant strain, a lower competence in the rhizosphere of A. caven was also shown, both in gnotobiotic in vitro cultures and in plant/soil microcosms.


Assuntos
Acacia/microbiologia , Arabidopsis/microbiologia , Cupriavidus/crescimento & desenvolvimento , Cupriavidus/metabolismo , Hidrocarbonetos Aromáticos/metabolismo , Acacia/metabolismo , Arabidopsis/metabolismo , Raízes de Plantas/metabolismo , Raízes de Plantas/microbiologia , Rizosfera
15.
Front Bioeng Biotechnol ; 10: 859600, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36072290

RESUMO

Gut metabolites are pivotal mediators of host-microbiome interactions and provide an important window on human physiology and disease. However, current methods to monitor gut metabolites rely on heavy and expensive technologies such as liquid chromatography-mass spectrometry (LC-MS). In that context, robust, fast, field-deployable, and cost-effective strategies for monitoring fecal metabolites would support large-scale functional studies and routine monitoring of metabolites biomarkers associated with pathological conditions. Living cells are an attractive option to engineer biosensors due to their ability to detect and process many environmental signals and their self-replicating nature. Here we optimized a workflow for feces processing that supports metabolite detection using bacterial biosensors. We show that simple centrifugation and filtration steps remove host microbes and support reproducible preparation of a physiological-derived media retaining important characteristics of human feces, such as matrix effects and endogenous metabolites. We measure the performance of bacterial biosensors for benzoate, lactate, anhydrotetracycline, and bile acids, and find that they are highly sensitive to fecal matrices. However, encapsulating the bacteria in hydrogel helps reduce this inhibitory effect. Sensitivity to matrix effects is biosensor-dependent but also varies between individuals, highlighting the need for case-by-case optimization for biosensors' operation in feces. Finally, by detecting endogenous bile acids, we demonstrate that bacterial biosensors could be used for future metabolite monitoring in feces. This work lays the foundation for the optimization and use of bacterial biosensors for fecal metabolites monitoring. In the future, our method could also allow rapid pre-prototyping of engineered bacteria designed to operate in the gut, with applications to in situ diagnostics and therapeutics.

16.
ACS Synth Biol ; 11(8): 2578-2588, 2022 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-35913043

RESUMO

Cell-free systems have great potential for delivering robust, inexpensive, and field-deployable biosensors. Many cell-free biosensors rely on transcription factors responding to small molecules, but their discovery and implementation still remain challenging. Here we report the engineering of PeroxiHUB, an optimized H2O2-centered sensing platform supporting cell-free detection of different metabolites. H2O2 is a central metabolite and a byproduct of numerous enzymatic reactions. PeroxiHUB uses enzymatic transducers to convert metabolites of interest into H2O2, enabling rapid reprogramming of sensor specificity using alternative transducers. We first screen several transcription factors and optimize OxyR for the transcriptional response to H2O2 in a cell-free system, highlighting the need for preincubation steps to obtain suitable signal-to-noise ratios. We then demonstrate modular detection of metabolites of clinical interest─lactate, sarcosine, and choline─using different transducers mined via a custom retrosynthesis workflow publicly available on the SynBioCAD Galaxy portal. We find that expressing the transducer during the preincubation step is crucial for optimal sensor operation. We then show that different reporters can be connected to PeroxiHUB, providing high adaptability for various applications. Finally, we demonstrate that a peroxiHUB lactate biosensor can detect endogenous levels of this metabolite in clinical samples. Given the wide range of enzymatic reactions producing H2O2, the PeroxiHUB platform will support cell-free detection of a large number of metabolites in a modular and scalable fashion.


Assuntos
Técnicas Biossensoriais , Peróxido de Hidrogênio , Sistema Livre de Células/metabolismo , Peróxido de Hidrogênio/metabolismo , Fatores de Transcrição/genética
17.
ACS Synth Biol ; 10(12): 3527-3536, 2021 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-34851606

RESUMO

Bacteria equipped with genetically encoded lactate biosensors are promising tools for biopharmaceutical production, diagnostics, and cellular therapies. However, many applications involve glucose-rich and anoxic environments, in which current whole-cell lactate biosensors show low performance. Here we engineer an optimized, synthetic lactate biosensor system by repurposing the natural LldPRD promoter regulated by the LldR transcriptional regulator. We removed glucose catabolite and anoxic repression by designing a hybrid promoter, containing LldR operators and tuned both regulator and reporter gene expressions to optimize biosensor signal-to-noise ratio. The resulting lactate biosensor, termed ALPaGA (A Lactate Promoter Operating in Glucose and Anoxia), can operate in glucose-rich, aerobic and anoxic conditions. We show that ALPaGA works reliably in the probiotic chassisEscherichia coliNissle 1917 and can detect endogenous l-lactate produced by 3D tumor spheroids with an improved dynamic range. In the future, the ALPaGA system could be used to monitor bioproduction processes and improve the specificity of engineered bacterial cancer therapies by restricting their activity to the lactate-rich microenvironment of solid tumors.


Assuntos
Técnicas Biossensoriais , Regulação Bacteriana da Expressão Gênica , Glucose , Humanos , Hipóxia , Ácido Láctico/metabolismo , Regiões Promotoras Genéticas/genética
18.
Nat Commun ; 12(1): 5216, 2021 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-34471137

RESUMO

Bacterial biosensors, or bactosensors, are promising agents for medical and environmental diagnostics. However, the lack of scalable frameworks to systematically program ligand detection limits their applications. Here we show how novel, clinically relevant sensing modalities can be introduced into bactosensors in a modular fashion. To do so, we have leveraged a synthetic receptor platform, termed EMeRALD (Engineered Modularized Receptors Activated via Ligand-induced Dimerization) which supports the modular assembly of sensing modules onto a high-performance, generic signaling scaffold controlling gene expression in E. coli. We apply EMeRALD to detect bile salts, a biomarker of liver dysfunction, by repurposing sensing modules from enteropathogenic Vibrio species. We improve the sensitivity and lower the limit-of-detection of the sensing module by directed evolution. We then engineer a colorimetric bactosensor detecting pathological bile salt levels in serum from patients having undergone liver transplant, providing an output detectable by the naked-eye. The EMeRALD technology enables functional exploration of natural sensing modules and rapid engineering of synthetic receptors for diagnostics, environmental monitoring, and control of therapeutic microbes.


Assuntos
Bactérias/metabolismo , Biomarcadores/metabolismo , Técnicas Biossensoriais , Proteínas de Transporte/metabolismo , Patologia Molecular/métodos , Bactérias/genética , Ácidos e Sais Biliares/sangue , Técnicas Biossensoriais/métodos , Proteínas de Transporte/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Regulação Bacteriana da Expressão Gênica , Humanos , Transplante de Fígado , Engenharia Metabólica/métodos , Sensibilidade e Especificidade , Alinhamento de Sequência , Vibrio , Vibrioses/diagnóstico
19.
Minerva Anestesiol ; 87(6): 663-674, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33591141

RESUMO

BACKGROUND: Optimal control of acute postoperative pain and prevention of chronic persistent pain in total hip arthroplasty (THA) remain a challenge. The main hypothesis was that peripheral nerve blocks improve postoperative analgesia. METHODS: Immediate postoperative pain (24 hours) was evaluated every hour in 510 patients using a verbal rating 11-point scale for patient self-reporting of pain (VRS-11). All patients received subarachnoid anesthesia (SA) and were randomly allocated in six groups: SA with morphine 0.1 (SA0.1) or 0.2 mg (SA0.2), fascia iliaca compartment block with dexamethasone 4 mg + levobupivacaine 0.375% 20 (FICB20) or 30 mL (FICB30), lateral femoral cutaneous nerve block with levobupivacaine 0.25% 5 mL (LFCNB) and FICB20+LFCNB. Standardized analgesia included intravenous metamizole magnesium, dexketoprofen and rescue with paracetamol and morphine, and/or regional rescue (FICB, LFCNB, femoral and sciatic nerve blocks). RESULTS: About 37.5% of patients had at least one episode of pain, 31.3% of them needed rescue analgesia while the remaining 6.2% did not request analgesia. There were no significant differences between the groups in paracetamol, morphine and rescue nerve blocks requirements. There was pain only in 5.4% of the total PACU pain records: 3.1% mild pain, 1.7% moderate pain and 0.6% severe pain. CONCLUSIONS: combined with a multimodal analgesic approach, infra-inguinal FICB and LFCNB did not improve immediate postoperative analgesia for THA in our hospital. Other options and longer-term studies should be more extensively investigated to determine the role of peripheral blocks in postoperative pain treatment protocols.


Assuntos
Analgesia , Artroplastia de Quadril , Bloqueio Nervoso , Analgésicos Opioides/uso terapêutico , Humanos , Dor Pós-Operatória/tratamento farmacológico , Dor Pós-Operatória/prevenção & controle
20.
J Clin Densitom ; 13(4): 467-9, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21029978

RESUMO

Shift workers have been reported to have an increased bone resorption. However, no existing evidence indicates lower bone mineral density (BMD) in this group. The objective of this study was to test the hypothesis that a rotating-shift work schedule is associated with low BMD and osteoporosis. We evaluated 70 postmenopausal nurses from the Naval Hospital in Concepcion, Chile. The participants were categorized according to the type of work schedule: 39 had a rotating shift and 31 were daytime workers. Medical history, a health examination, a questionnaire on health-related behaviors and biochemical determinations, and BMD examination were obtained for all participants. When comparing the 2 groups, the rotating-shift workers had lower BMD in the lumbar spine (L1-L4: 0.957 ± 0.15 vs 1.104 ± 0.13; p<0.05) and lower bone density in both femoral neck bones (right: 0.936 ± 0.17 vs 1.06 ± 0.12; p<0.05 and left: 0.956 ± 0.19 vs 1.05 ± 0.12; p<0.05). Additionally, the T-scores for 10 (25.6%) of the rotating-shift workers indicated osteoporosis at lumbar spine (T-score>-2.5). No evidence of osteoporosis was found for daytime workers. When comparing the 2 groups, the rotating-shift workers had a higher prevalence of osteopenia (T-score=-1.0 to -2.5) than the daytime workers: 46.2% vs 35.5%, respectively. We found significant evidence that rotating-shift workers have lower BMD in the trabecular and cortical bones, thus suggesting that this type of work may be a risk factor for osteoporosis. Because this is the first time that this osteoporosis risk factor has been reported, the association needs to be replicated and confirmed in other settings.


Assuntos
Densidade Óssea , Osteoporose Pós-Menopausa/epidemiologia , Tolerância ao Trabalho Programado , Absorciometria de Fóton , Biomarcadores/sangue , Feminino , Humanos , Pessoa de Meia-Idade , Fatores de Risco , Inquéritos e Questionários
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