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1.
Urol Int ; 107(8): 778-784, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37544300

RESUMO

INTRODUCTION: Acute lower uncomplicated urinary tract infection (uUTI) affects a large proportion of women. Increased antimicrobial resistance has created an urgent need for novel therapeutics and the phytotherapeutic drug BNO 1045 (Canephron® N) has previously been shown to be noninferior to standard antimicrobial stewardship. This sub-analysis from a randomized, double-blind, controlled phase III noninferiority clinical trial using BNO 1045 versus fosfomycin to treat uUTI aimed to determine how urine cytokine levels are altered by the two different treatments. METHODS: Urine samples from a predefined subset of women diagnosed with uUTI (18-70 years) and treated with BNO 1045 (n = 58) or fosfomycin (n = 69) were analyzed for urine levels of IL-6 and IL-8, using analyte-to-creatinine ratios. RESULTS: BNO 1045 treatment showed similar effects to fosfomycin treatment in reducing both urine IL-6 and IL-8 levels. Mean IL-6 and IL-8 levels were markedly reduced in all patients regardless of treatment. BNO 1045 treatment decreased urine IL-8 significantly (p = 0.0142) and showed a trend toward reduction of urine IL-6 (p = 0.0551). Fosfomycin treatment reduced both IL-6 and IL-8 levels significantly (p = 0.0038, <0.0001 respectively). CONCLUSION: BNO 1045 is, in addition to reducing symptoms, comparable to fosfomycin treatment in reducing the local inflammatory response associated with uUTI.


Assuntos
Fosfomicina , Infecções Urinárias , Humanos , Feminino , Fosfomicina/uso terapêutico , Interleucina-8 , Interleucina-6 , Infecções Urinárias/tratamento farmacológico , Fitoterapia , Antibacterianos/uso terapêutico
2.
J Vis Exp ; (196)2023 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-37427934

RESUMO

Cell polarity is a macroscopic phenomenon established by a collection of spatially concentrated molecules and structures that culminate in the emergence of specialized domains at the subcellular level. It is associated with developing asymmetric morphological structures that underlie key biological functions such as cell division, growth, and migration. In addition, the disruption of cell polarity has been linked to tissue-related disorders such as cancer and gastric dysplasia. Current methods to evaluate the spatiotemporal dynamics of fluorescent reporters in individual polarized cells often involve manual steps to trace a midline along the cells' major axis, which is time consuming and prone to strong biases. Furthermore, although ratiometric analysis can correct the uneven distribution of reporter molecules using two fluorescence channels, background subtraction techniques are frequently arbitrary and lack statistical support. This manuscript introduces a novel computational pipeline to automate and quantify the spatiotemporal behavior of single cells using a model of cell polarity: pollen tube/root hair growth and cytosolic ion dynamics. A three-step algorithm was developed to process ratiometric images and extract a quantitative representation of intracellular dynamics and growth. The first step segments the cell from the background, producing a binary mask through a thresholding technique in the pixel intensity space. The second step traces a path through the midline of the cell through a skeletonization operation. Finally, the third step provides the processed data as a ratiometric timelapse and yields a ratiometric kymograph (i.e., a 1D spatial profile through time). Data from ratiometric images acquired with genetically encoded fluorescent reporters from growing pollen tubes were used to benchmark the method. This pipeline allows for faster, less biased, and more accurate representation of the spatiotemporal dynamics along the midline of polarized cells, thus advancing the quantitative toolkit available to investigate cell polarity. The AMEBaS Python source code is available at: https://github.com/badain/amebas.git.


Assuntos
Polaridade Celular , Software , Imagem com Lapso de Tempo , Algoritmos , Tubo Polínico , Corantes
3.
Pathogens ; 13(1)2023 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-38251349

RESUMO

Innate immunity is essential for the anti-microbial defense, but excessive immune activation may cause severe disease. In this study, immunotherapy was shown to prevent excessive innate immune activation and restore the anti-bacterial defense. E. coli-infected Asc-/- mice develop severe acute cystitis, defined by IL-1 hyper-activation, high bacterial counts, and extensive tissue pathology. Here, the interleukin-1 receptor antagonist (IL-1RA), which inhibits IL-1 hyper-activation in acute cystitis, was identified as a more potent inhibitor of inflammation and NK1R- and substance P-dependent pain than cefotaxime. Furthermore, IL-1RA treatment inhibited the excessive innate immune activation in the kidneys of infected Irf3-/- mice and restored tissue integrity. Unexpectedly, IL-1RA also accelerated bacterial clearance from infected bladders and kidneys, including antibiotic-resistant E. coli, where cefotaxime treatment was inefficient. The results suggest that by targeting the IL-1 response, control of the innate immune response to infection may be regained, with highly favorable treatment outcomes, including infections caused by antibiotic-resistant strains.

4.
IEEE Sens J ; 22(19): 18437-18445, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36416744

RESUMO

The development of a cost-efficient device to rapidly detect pandemic viruses is paramount. Hence, an innovative and scalable synthesis of metal nanoparticles followed by its usage for rapid detection of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been reported in this work. The simple synthesis of metal nanoparticles utilizing tin as a solid-state reusable reducing agent is used for the SARS-CoV-2 ribonucleic acid (RNA) detection. Moreover, the solid-state reduction process occurs faster and leads to the enhanced formation of silver and gold nanoparticles (AuNPs) with voltage. By adding tin as a solid-state reducing agent with the precursor, the nanoparticles are formed within 30 s. This synthesis method can be easily scaled up for a commercially viable process to obtain different-sized metal nanoparticles. This is the first disclosure of the usage of tin as a reusable solid-state reducing agent for metal nanoparticle synthesis. An electronic device, consisting of AuNPs functionalized with a deoxyribonucleic acid (DNA)-based aptamer, can detect SARS-CoV-2 RNA in less than 5 min. With an increase in SARS-CoV-2 variants, such as Delta and Omicron, the detection device could be used for identifying the nucleic acids of the COVID-19 variants by modifying the aptamer sequence. The reported work overcomes the drawbacks of complex instrumentation, trained labor, and increased turnaround time.

5.
Curr Opin Cell Biol ; 77: 102113, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35809387

RESUMO

Physiological oscillations (or rhythms) pervade all spatiotemporal scales of biological organization, either because they perform critical functions or simply because they can arise spontaneously and may be difficult to prevent. Regardless of the case, they reflect regulatory relationships between control points of a given system and offer insights as read-outs of the concerted regulation of a myriad of biological processes. Here we review recent advances in understanding ultradian oscillations (period < 24h) in plant cells, with a special focus on single-cell oscillations. Ion channels are at the center stage due to their involvement in electrical/excitabile phenomena associated with oscillations and cell-cell communication. We highlight the importance of quantitative approaches to measure oscillations in appropriate physiological conditions, which are essential strategies to deal with the complexity of biological rhythms. Future development of optogenetics techniques in plants will further boost research on the role of membrane potential in oscillations and waves across multiple cell types.


Assuntos
Comunicação Celular , Células Vegetais
6.
Commun Biol ; 5(1): 6, 2022 01 10.
Artigo em Inglês | MEDLINE | ID: mdl-35013546

RESUMO

It is unclear whether West Nile virus (WNV) circulates endemically in Portugal. Despite the country's adequate climate for transmission, Portugal has only reported four human WNV infections so far. We performed a review of WNV-related data (1966-2020), explored mosquito (2016-2019) and land type distributions (1992-2019), and used climate data (1981-2019) to estimate WNV transmission suitability in Portugal. Serological and molecular evidence of WNV circulation from animals and vectors was largely restricted to the south. Land type and climate-driven transmission suitability distributions, but not the distribution of WNV-capable vectors, were compatible with the North-South divide present in serological and molecular evidence of WNV circulation. Our study offers a comprehensive, data-informed perspective and review on the past epidemiology, surveillance and climate-driven transmission suitability of WNV in Portugal, highlighting the south as a subregion of importance. Given the recent WNV outbreaks across Europe, our results support a timely change towards local, active surveillance.


Assuntos
Distribuição Animal , Clima , Tempo (Meteorologia) , Febre do Nilo Ocidental/transmissão , Vírus do Nilo Ocidental/isolamento & purificação , Animais , Culicidae/fisiologia , Humanos , Mosquitos Vetores/fisiologia , Portugal , Estações do Ano , Especificidade da Espécie , Vírus do Nilo Ocidental/fisiologia
7.
Eur Urol Open Sci ; 31: 49-58, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34467240

RESUMO

BACKGROUND: Patients with bladder pain syndrome experience debilitating pain and extreme frequency of urination. Numerous therapeutic approaches have been tested, but as the molecular basis of disease has remained unclear, specific therapies are not available. OBJECTIVE: Recently, a systematic gene deletion strategy identified interleukin-1 (IL-1) hyperactivation as a cause of severe cystitis in a murine model. Treatment with an IL-1 receptor antagonist (IL-1RA) restored health in genetically susceptible mice, linking IL-1-dependent inflammation to pain and pathology in the bladder mucosa. The study objective was to investigate whether IL-1RA treatment might be beneficial in patients with bladder pain syndrome. DESIGN SETTING AND PARTICIPANTS: Patients diagnosed with bladder pain syndrome were invited to participate and subjected to daily IL-1RA injections for 1 wk, followed by a treatment break. Patients with other urological disorders accompanied by pain were included as controls. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: When symptoms returned, treatment was resumed and responding patients were maintained on treatment long term, with individualized dosing regimens. Symptom scores were recorded and molecular effects were quantified by neuropeptide and gene expression analysis. DNA samples were subjected to exome genotyping. RESULTS AND LIMITATIONS: IL-1RA treatment reduced bladder pain and the frequency of urination in 13/17 patients (p < 0.001). Substance P levels in urine were lowered, and responders returned to a more normal lifestyle. Neuroinflammatory-dependent and IL-1-dependent gene networks were inhibited, as well as regulators of innate immunity. Genotyping revealed disease-associated IL1R1, NLRP3, and IL1RN DNA sequence variants in the responders. Controls did not benefit from IL-1RA treatment, except for one patent with cystitis cystica. CONCLUSIONS: In this clinical study, IL-1RA treatment is proposed to reduce chronic bladder pain, immediately and in the long term. Despite the limited number of study patients, the potent acute effect and lasting symptom relief indicate that this therapeutic approach may be worth exploring in controlled clinical trials. PATIENT SUMMARY: Treatment with an interleukin-1 (IL-1) receptor antagonist is proposed for treating bladder pain syndrome, as it can result in symptom relief and increase quality of life. Reduced neuroinflammation and IL-1 signaling provided molecular evidence of the treatment effects.

8.
Bio Protoc ; 11(14): e4084, 2021 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-34395723

RESUMO

Ion-specific probes and fluorescent indicators have been key in establishing the role of ion signaling, namely calcium, protons, and anions, in plant development, providing a robust approach for monitoring spatiotemporal changes in intracellular ion dynamics. The integration of protons/pH in signaling mechanisms is especially important as reports of their biological functions continue to expand; however, attaining quantitative estimates with high spatiotemporal resolution in single cells poses a major research challenge. Here, we detail the use of the genetically encoded pH-sensitive pHluorin reporter expressed in Arabidopsis thaliana pollen tubes to assess cytosolic measurements with calibration to provide actual pH values. This technique enabled us to identify critical phenotypes and establish the importance of tip-focused pH gradient for pollen tube growth, although it can be adapted to other experimental systems.

9.
Nat Commun ; 12(1): 3427, 2021 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-34103518

RESUMO

Partially unfolded alpha-lactalbumin forms the oleic acid complex HAMLET, with potent tumoricidal activity. Here we define a peptide-based molecular approach for targeting and killing tumor cells, and evidence of its clinical potential (ClinicalTrials.gov NCT03560479). A 39-residue alpha-helical peptide from alpha-lactalbumin is shown to gain lethality for tumor cells by forming oleic acid complexes (alpha1-oleate). Nuclear magnetic resonance measurements and computational simulations reveal a lipid core surrounded by conformationally fluid, alpha-helical peptide motifs. In a single center, placebo controlled, double blinded Phase I/II interventional clinical trial of non-muscle invasive bladder cancer, all primary end points of safety and efficacy of alpha1-oleate treatment are reached, as evaluated in an interim analysis. Intra-vesical instillations of alpha1-oleate triggers massive shedding of tumor cells and the tumor size is reduced but no drug-related side effects are detected (primary endpoints). Shed cells contain alpha1-oleate, treated tumors show evidence of apoptosis and the expression of cancer-related genes is inhibited (secondary endpoints). The results are especially encouraging for bladder cancer, where therapeutic failures and high recurrence rates create a great, unmet medical need.


Assuntos
Peptídeos/química , Peptídeos/uso terapêutico , Neoplasias da Bexiga Urinária/tratamento farmacológico , Sequência de Aminoácidos , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Endocitose/efeitos dos fármacos , Determinação de Ponto Final , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Ácidos Oleicos/química , Peptídeos/farmacologia , Placebos , Conformação Proteica , Espectroscopia de Prótons por Ressonância Magnética , Termodinâmica , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/patologia
10.
Artigo em Inglês | MEDLINE | ID: mdl-34007909

RESUMO

BACKGROUND: The exploration of Advanced Practiced Radiation Therapists (APRTs) development in Singapore started in 2011. This study aims to provide an overview of the development of the APRT roles, and to discuss the approaches used to develop and implement these roles in Singapore. MATERIALS AND METHODS: A mixed methods approach was used in the development of the APRT program. A literature review was carried out to define the APRT scope of practice and core responsibilities. A competency and assessment framework were setup to assess the core competency areas. With this framework, a structured 1-year residency training program was developed. RESULTS: The scope of practice and core responsibilities of APRTs were defined with five proposed advanced practice profiles being successfully validated. A competency framework was set up to assess the core competency domains: clinical, technical and professional competencies, research, education and leadership. A 4-point scoring system was developed for the competency assessment based on two criteria; the frequency with which RTTs would demonstrate competency, and the ability of performing the task competently. A 1-year structured APRT residency program was developed and implemented. The programme consisted of structured lectures, and clinical practice-based modules where APRT residents receive structured mentoring under a mentorship program. CONCLUSION: The APRT program in Singapore employed an evidence-based implementation process that tested the feasibility of a new practice model. Multidisciplinary involvements, mentorship and clinical training were important factors for the success of the APRT program.

11.
Nat Biotechnol ; 39(6): 754-764, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33574609

RESUMO

Is the oncogene MYC upregulated or hyperactive? In the majority of human cancers, finding agents that target c-MYC has proved difficult. Here we report specific bacterial effector molecules that inhibit cellular MYC (c-MYC) in human cells. We show that uropathogenic Escherichia coli (UPEC) degrade the c-MYC protein and attenuate MYC expression in both human cells and animal tissues. c-MYC protein was rapidly degraded by both cell-free bacterial lysates and the purified bacterial protease Lon. In mice, intravesical or peroral delivery of Lon protease delayed tumor progression and increased survival in MYC-dependent bladder and colon cancer models, respectively. These results suggest that bacteria have evolved strategies to control c-MYC tissue levels in the host and that the Lon protease shows promise for therapeutic targeting of c-MYC in cancer.


Assuntos
Neoplasias do Colo/patologia , Genes myc , Neoplasias da Bexiga Urinária/patologia , Escherichia coli Uropatogênica/enzimologia , Doença Aguda , Animais , Modelos Animais de Doenças , Infecções por Escherichia coli/patologia , Deleção de Genes , Camundongos , Nefrite/genética
12.
BMC Med ; 19(1): 19, 2021 01 12.
Artigo em Inglês | MEDLINE | ID: mdl-33430856

RESUMO

BACKGROUND: Cross-reactivity to SARS-CoV-2 from exposure to endemic human coronaviruses (eHCoV) is gaining increasing attention as a possible driver of both protection against infection and COVID-19 severity. Here we explore the potential role of cross-reactivity induced by eHCoVs on age-specific COVID-19 severity in a mathematical model of eHCoV and SARS-CoV-2 transmission. METHODS: We use an individual-based model, calibrated to prior knowledge of eHCoV dynamics, to fully track individual histories of exposure to eHCoVs. We also model the emergent dynamics of SARS-CoV-2 and the risk of hospitalisation upon infection. RESULTS: We hypothesise that primary exposure with any eHCoV confers temporary cross-protection against severe SARS-CoV-2 infection, while life-long re-exposure to the same eHCoV diminishes cross-protection, and increases the potential for disease severity. We show numerically that our proposed mechanism can explain age patterns of COVID-19 hospitalisation in EU/EEA countries and the UK. We further show that some of the observed variation in health care capacity and testing efforts is compatible with country-specific differences in hospitalisation rates under this model. CONCLUSIONS: This study provides a "proof of possibility" for certain biological and epidemiological mechanisms that could potentially drive COVID-19-related variation across age groups. Our findings call for further research on the role of cross-reactivity to eHCoVs and highlight data interpretation challenges arising from health care capacity and SARS-CoV-2 testing.


Assuntos
COVID-19 , Infecções por Coronavirus , Proteção Cruzada/imunologia , Reações Cruzadas/imunologia , SARS-CoV-2/imunologia , Fatores Etários , COVID-19/epidemiologia , COVID-19/imunologia , COVID-19/fisiopatologia , Coronavirus/classificação , Coronavirus/imunologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/terapia , Doenças Endêmicas , Hospitalização/estatística & dados numéricos , Humanos , Imunidade Heteróloga/imunologia , Modelagem Computacional Específica para o Paciente , Índice de Gravidade de Doença
13.
New Phytol ; 230(6): 2292-2310, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33455006

RESUMO

Whereas the role of calcium ions (Ca2+ ) in plant signaling is well studied, the physiological significance of pH-changes remains largely undefined. Here we developed CapHensor, an optimized dual-reporter for simultaneous Ca2+ and pH ratio-imaging and studied signaling events in pollen tubes (PTs), guard cells (GCs), and mesophyll cells (MCs). Monitoring spatio-temporal relationships between membrane voltage, Ca2+ - and pH-dynamics revealed interconnections previously not described. In tobacco PTs, we demonstrated Ca2+ -dynamics lag behind pH-dynamics during oscillatory growth, and pH correlates more with growth than Ca2+ . In GCs, we demonstrated abscisic acid (ABA) to initiate stomatal closure via rapid cytosolic alkalization followed by Ca2+ elevation. Preventing the alkalization blocked GC ABA-responses and even opened stomata in the presence of ABA, disclosing an important pH-dependent GC signaling node. In MCs, a flg22-induced membrane depolarization preceded Ca2+ -increases and cytosolic acidification by c. 2 min, suggesting a Ca2+ /pH-independent early pathogen signaling step. Imaging Ca2+ and pH resolved similar cytosol and nuclear signals and demonstrated flg22, but not ABA and hydrogen peroxide to initiate rapid membrane voltage-, Ca2+ - and pH-responses. We propose close interrelation in Ca2+ - and pH-signaling that is cell type- and stimulus-specific and the pH having crucial roles in regulating PT growth and stomata movement.


Assuntos
Cálcio , Estômatos de Plantas/fisiologia , Transdução de Sinais , Ácido Abscísico , Citosol/metabolismo , Concentração de Íons de Hidrogênio
14.
J Comput Aided Mol Des ; 34(11): 1157-1169, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32648151

RESUMO

Experimental 3D structures of calcium channels with phenylalkylamines (PAAs) provide basis for further analysis of atomic mechanisms of these important cardiovascular drugs. In the crystal structure of the engineered calcium channel CavAb with Br-verapamil and in the cryo-EM structure of the Cav1.1 channel with verapamil, the ligands bind in the inner pore. However, there are significant differences between these structures. In the crystal structure the ligand ammonium group is much closer to the ion in the selectivity-filter region Site 3, which is most proximal to the inner pore, than in the cryo-EM structure. Here we used Monte Carlo energy minimizations to dock PAAs in calcium channels. Our computations suggest that in the crystal structure Site 3 is occupied by a water molecule rather than by a calcium ion. Analysis of the published electron density map does not rule out this possibility. In the cryo-EM structures the ammonium group of verapamil is shifted from the calcium ion in Site 3 either along the pore axis, towards the cytoplasm or away from the axis. Our unbiased docking reproduced these binding modes. However, in the cryo-EM structures detergent and lipid molecules interact with verapamil. When we removed these molecules, the nitrile group of verapamil bound to the calcium ion in Site 3. Models of Cav1.2 with different PAAs suggest similar binding modes and direct contacts of the ligands electronegative atoms with the calcium ion in Site 3. Such interactions explain paradoxes in structure-activity relationships of PAAs.


Assuntos
Bloqueadores dos Canais de Cálcio/química , Canais de Cálcio/metabolismo , Verapamil/química , Sequência de Aminoácidos , Sítios de Ligação , Cálcio/química , Cristalização , Ligantes , Simulação de Acoplamento Molecular , Método de Monte Carlo , Relação Estrutura-Atividade
15.
Methods Mol Biol ; 2160: 201-210, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32529438

RESUMO

Conspicuous intracellular gradients manifest and/or drive intracellular polarity in pollen tubes. However, quantifying these gradients raises multiple technical challenges. Here we present a sensible computational protocol to analyze gradients in growing pollen tubes and to filter nonrepresentative time points. As an example, we use imaging data from pollen tubes expressing a genetically encoded ratiometric Ca2+ probe, Yellow CaMeleon 3.6, from which a kymograph is extracted. The tip of the pollen tube is detected with CHUKNORRIS, our previously published methodology, allowing the reconstruction of the intracellular gradient through time. Statistically confounding time points, such as growth arrest where gradients are highly oscillatory, are filtered out and a mean spatial profile is estimated with a local polynomial regression method. Finally, we estimate the gradient slope by the linear portion of the decay in mean fluorescence, offering a quantitative method to detect phenotypes of gradient steepness, location, intensity, and variability. The data manipulation protocol proposed can be achieved in a simple and efficient manner using the statistical programming language R, opening paths to perform high-throughput spatiotemporal phenotyping of intracellular gradients in apically growing cells.


Assuntos
Análise do Fluxo Metabólico/instrumentação , Tubo Polínico/metabolismo , Arabidopsis , Cálcio/metabolismo , Polaridade Celular , Quimografia/métodos , Análise do Fluxo Metabólico/métodos , Microscopia de Fluorescência/métodos , Tubo Polínico/citologia , Software
16.
PLoS Pathog ; 15(6): e1007671, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31181116

RESUMO

Pathogens rely on a complex virulence gene repertoire to successfully attack their hosts. We were therefore surprised to find that a single fimbrial gene reconstitution can return the virulence-attenuated commensal strain Escherichia coli 83972 to virulence, defined by a disease phenotype in human hosts. E. coli 83972pap stably reprogrammed host gene expression, by activating an acute pyelonephritis-associated, IRF7-dependent gene network. The PapG protein was internalized by human kidney cells and served as a transcriptional agonist of IRF-7, IFN-ß and MYC, suggesting direct involvement of the fimbrial adhesin in this process. IRF-7 was further identified as a potent upstream regulator (-log (p-value) = 61), consistent with the effects in inoculated patients. In contrast, E. coli 83972fim transiently attenuated overall gene expression in human hosts, enhancing the effects of E. coli 83972. The inhibition of RNA processing and ribosomal assembly indicated a homeostatic rather than a pathogenic end-point. In parallel, the expression of specific ion channels and neuropeptide gene networks was transiently enhanced, in a FimH-dependent manner. The studies were performed to establish protective asymptomatic bacteriuria in human hosts and the reconstituted E. coli 83972 variants were developed to improve bacterial fitness for the human urinary tract. Unexpectedly, P fimbriae were able to drive a disease response, suggesting that like oncogene addiction in cancer, pathogens may be addicted to single super-virulence factors.


Assuntos
Adesinas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Proteínas de Fímbrias/metabolismo , Fímbrias Bacterianas/metabolismo , Adesinas de Escherichia coli/genética , Linhagem Celular , Escherichia coli/genética , Escherichia coli/patogenicidade , Feminino , Proteínas de Fímbrias/genética , Fímbrias Bacterianas/genética , Humanos , Fator Regulador 7 de Interferon/metabolismo , Interferon beta/metabolismo , Rim/metabolismo , Rim/microbiologia , Proteínas Proto-Oncogênicas c-myc/metabolismo
17.
Sci Rep ; 8(1): 11015, 2018 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-30030504

RESUMO

The nervous system is engaged by infection, indirectly through inflammatory cascades or directly, by bacterial attack on nerve cells. Here we identify a neuro-epithelial activation loop that participates in the control of mucosal inflammation and pain in acute cystitis. We show that infection activates Neurokinin-1 receptor (NK1R) and Substance P (SP) expression in nerve cells and bladder epithelial cells in vitro and in vivo in the urinary bladder mucosa. Specific innate immune response genes regulated this mucosal response, and single gene deletions resulted either in protection (Tlr4-/- and Il1b-/- mice) or in accentuated bladder pathology (Asc-/- and Nlrp3-/- mice), compared to controls. NK1R/SP expression was lower in Tlr4-/- and Il1b-/- mice than in C56BL/6WT controls but in Asc-/- and Nlrp3-/- mice, NK1R over-activation accompanied the exaggerated disease phenotype, due, in part to transcriptional de-repression of Tacr1. Pharmacologic NK1R inhibitors attenuated acute cystitis in susceptible mice, supporting a role in disease pathogenesis. Clinical relevance was suggested by elevated urine SP levels in patients with acute cystitis, compared to patients with asymptomatic bacteriuria identifying NK1R/SP as potential therapeutic targets. We propose that NK1R and SP influence the severity of acute cystitis through a neuro-epithelial activation loop that controls pain and mucosal inflammation.


Assuntos
Cistite/patologia , Células Neuroepiteliais/fisiologia , Receptores da Neurocinina-1/metabolismo , Adulto , Animais , Cistite/imunologia , Feminino , Expressão Gênica/efeitos dos fármacos , Humanos , Imunidade nas Mucosas/fisiologia , Inflamação/patologia , Interleucina-1beta/genética , Macrófagos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mucosite/patologia , Músculo Liso/metabolismo , Células Neuroepiteliais/metabolismo , Neutrófilos , Dor/patologia , Substância P/metabolismo , Receptor 4 Toll-Like/genética , Bexiga Urinária/metabolismo , Bexiga Urinária/patologia
18.
Nature ; 549(7670): 91-95, 2017 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-28737761

RESUMO

Glutamate receptors are well characterized channels that mediate cell-to-cell communication during neurotransmission in animals, but their functional role in organisms without a nervous system remains unclear. In plants, genes of the GLUTAMATE RECEPTOR-LIKE (GLR) family have been implicated in defence against pathogens, reproduction, control of stomata aperture and light signal transduction. However, the large number of GLR genes present in angiosperm genomes (20 to 70) has prevented the observation of strong phenotypes in loss-of-function mutants. Here we show that in the basal land plant Physcomitrella patens, mutation of the GLR genes GLR1 and GLR2 causes failure of sperm cells to target the female reproductive organs. In addition, we show that GLR genes encode non-selective Ca2+-permeable channels that can regulate cytoplasmic Ca2+ and are needed to induce the expression of a BELL1-like transcription factor essential for zygote development. Our work reveals functions for GLR channels in sperm chemotaxis and transcriptional regulation. Sperm chemotaxis is essential for fertilization in both animals and early land plants such as bryophytes and pteridophytes. Therefore, our results suggest that ionotropic glutamate receptors may have been conserved throughout plant evolution to mediate cell-to-cell communication during sexual reproduction.


Assuntos
Bryopsida/metabolismo , Quimiotaxia , Receptores Ionotrópicos de Glutamato/metabolismo , Bryopsida/embriologia , Bryopsida/genética , Cálcio/metabolismo , Comunicação Celular/genética , Quimiotaxia/genética , Regulação da Expressão Gênica , Genes Essenciais , Mutação , Receptores Ionotrópicos de Glutamato/genética , Reprodução/genética , Fatores de Transcrição/metabolismo , Transcrição Gênica , Zigoto/metabolismo
19.
Mater Sci Eng C Mater Biol Appl ; 78: 1231-1245, 2017 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-28575962

RESUMO

Sensor technology for the rapid detection of the analytes with high sensitivity and selectivity has several challenges. Despite the challenges, colorimetric sensors have been widely accepted for its high sensitive and selective response towards various analytes. In this review, colorimetric sensors for the detection of biomolecules like protein, DNA, pathogen and chemical compounds like heavy metal ions, toxic gases and organic compounds have been elaborately discussed. The visible sensing mechanism based on Surface Plasmon Resonance (SPR) using metal nanoparticles like Au, Ag, thin film interference using SiO2 and colorimetric array-based technique have been highlighted. The optical property of metal nanoparticles enables a visual color change during its interaction with the analytes owing to the dispersion and aggregation of nanoparticles. Recently, colorimetric changes using silica substrate for detection of protein and small molecules by thin film interference as a visible sensing mechanism has been developed without the usage of fluorescent or radioisotopes labels. Multilayer of biomaterials were used as a platform where reflection and interference of scattering light occur due to which color change happens leading to rapid sensing. Colorimetric array-based technique for the detection of organic compounds using chemoresponsive dyes has also been focused wherein the interaction of the analytes with the substrate coated with chemoresponsive dyes gives colorimetric change.


Assuntos
Colorimetria , Ouro , Nanopartículas Metálicas , Dióxido de Silício , Ressonância de Plasmônio de Superfície
20.
Mater Sci Eng C Mater Biol Appl ; 75: 393-401, 2017 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-28415477

RESUMO

This work demonstrates the effective surface functionalization of Ag, Au and bimetallic Ag-Au nanoparticles using l-histidine for colorimetric detection of dopamine (DA) which plays majorly in recognizing the neurological disorder. l-Histidine (l-His) capped Ag, Au, and bimetallic Ag-Au nanoparticles are characterized using physico-chemical techniques. The optical behaviour of nanoparticles has been analysed at various time intervals using UV-Vis absorption spectroscopy. FT-IR results provide the evidence of chemical bonding between l-histidine and metal nanoparticles. Its structure with the capping of l-His was clearly shown in HR-TEM images. The average size of nanoparticles has calculated from TEM image fringes are 11nm, 5nm and 6.5nm respectively, matches with crystals size calculated from X-ray diffraction pattern. Enhanced optical nature of nanoparticles provides the best platform to develop a colorimetric-based biosensor for DA detection. After addition of DA, a rapid colour change has been noted in colloids of nanoparticles. The substantial changes in absorbance and λmax in metal nanoparticles respect to DA concentration have been observed and formulated. This is one of the successive methods for trace level determination of DA and will be going to a significant material for designing biosensor to determine DA in real extracellular body fluids.


Assuntos
Técnicas Biossensoriais/métodos , Dopamina/análise , Ouro/química , Histidina/química , Nanopartículas Metálicas/química , Prata/química , Materiais Revestidos Biocompatíveis , Espectrofotometria Ultravioleta
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