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1.
Mol Biol Cell ; 35(4): ar47, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38354034

RESUMO

Neuronal growth cones sense a variety of cues including chemical and mechanical ones to establish functional connections during nervous system development. Substrate-cytoskeletal coupling is an established model for adhesion-mediated growth cone advance; however, the detailed molecular and biophysical mechanisms underlying the mechanosensing and mechanotransduction process remain unclear. Here, we adapted a motor-clutch model to better understand the changes in clutch and cytoskeletal dynamics, traction forces, and substrate deformation when a growth cone interacts with adhesive substrates of different stiffnesses. Model parameters were optimized using experimental data from Aplysia growth cones probed with force-calibrated glass microneedles. We included a reinforcement mechanism at both motor and clutch level. Furthermore, we added a threshold for retrograde F-actin flow that indicates when the growth cone is strongly coupled to the substrate. Our modeling results are in strong agreement with experimental data with respect to the substrate deformation and the latency time after which substrate-cytoskeletal coupling is strong enough for the growth cone to advance. Our simulations show that it takes the shortest time to achieve strong coupling when substrate stiffness was low at 4 pN/nm. Taken together, these results suggest that Aplysia growth cones respond faster and more efficiently to soft than stiff substrates.


Assuntos
Cones de Crescimento , Mecanotransdução Celular , Cones de Crescimento/metabolismo , Actinas/metabolismo , Citoesqueleto/metabolismo , Células Fotorreceptoras Retinianas Cones
2.
Crit Rev Microbiol ; 49(3): 414-434, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-35574602

RESUMO

Clostridioides difficile infection (CDI) is a life-threatening disease caused by the Gram-positive, opportunistic intestinal pathogen C. difficile. Despite the availability of antimicrobial drugs to treat CDI, such as vancomycin, metronidazole, and fidaxomicin, recurrence of infection remains a significant clinical challenge. The use of live commensal microorganisms, or probiotics, is one of the most investigated non-antibiotic therapeutic options to balance gastrointestinal (GI) microbiota and subsequently tackle dysbiosis. In this review, we will discuss major commensal probiotic strains that have the potential to prevent and/or treat CDI and its recurrence, reassess the efficacy of probiotics supplementation as a CDI intervention, delve into lessons learned from probiotic modulation of the immune system, explore avenues like genome-scale metabolic network reconstructions, genome sequencing, and multi-omics to identify novel strains and understand their functionality, and discuss the current regulatory framework, challenges, and future directions.


Assuntos
Clostridioides difficile , Infecções por Clostridium , Probióticos , Humanos , Antibacterianos/uso terapêutico , Clostridioides difficile/genética , Clostridioides , Vancomicina/uso terapêutico , Infecções por Clostridium/tratamento farmacológico , Infecções por Clostridium/prevenção & controle , Probióticos/uso terapêutico
3.
Int J Appl Comput Math ; 7(3): 67, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33898652

RESUMO

To understand the dynamics of COVID-19 in Nigeria, a mathematical model which incorporates the key compartments and parameters regarding COVID-19 in Nigeria is formulated. The basic reproduction number is obtained which is then used to analyze the stability of the disease-free equilibrium solution of the model. The model is calibrated using data obtained from Nigeria Centre for Disease Control and key parameters of the model are estimated. Sensitivity analysis is carried out to investigate the influence of the parameters in curtailing the disease. Using Pontryagin's maximum principle, time-dependent intervention strategies are optimized in order to suppress the transmission of the virus. Numerical simulations are then used to explore various optimal control solutions involving single and multiple controls. Our results suggest that strict intervention effort is required for quick suppression of the disease.

4.
J Glob Antimicrob Resist ; 21: 154-161, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-31622683

RESUMO

OBJECTIVES: The rapid emergence of hypervirulent Clostridium difficile (C. difficile) isolates and the paucity of effective anti-clostridial antibiotics call for extensive research to identify new treatment options. This study aimed to test the anti-clostridial activity of bioactive extracts of turmeric, which is a natural herb widely known for its profound medicinal properties. METHODS: The MICs of turmeric derivatives were determined against 27 C. difficile strains, including hypervirulent (BI/NAP1/027) and clinical toxigenic isolates. Additionally, their ability to inhibit C. difficile toxin production and spore formation was investigated. Furthermore, the safety profiles of turmeric derivatives regarding their effects on human gut microflora - such as Bacteroides, Lactobacillus and Bifidobacterium - were evaluated. RESULTS: Curcuminoids, the major phytoconstituents of turmeric - including curcumin, demethoxycurcumin and bisdemethoxycurcumin - inhibited growth of C. difficile at concentrations ranging from 4 to 32µg/mL. Additionally, curcuminoids showed no negative effect on major populating species of the human gut. Curcumin was more effective than fidaxomicin in inhibiting C. difficile toxin production, but less so in inhibiting spore formation. CONCLUSION: The findings suggest that curcumin has potential as an anti-clostridial agent. More work is needed to further investigate the efficacy of curcumin as a stand-alone drug or as a supplement of current drugs of choice, as it has no antagonistic activities but might overcome their drawbacks.


Assuntos
Anti-Infecciosos , Clostridioides difficile , Curcumina , Antibacterianos/farmacologia , Curcumina/farmacologia , Humanos , Testes de Sensibilidade Microbiana
5.
Physiol Rep ; 7(6): e14030, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30912296

RESUMO

This study characterized the effects of regular green tea (GT) and hot water (HW) ingestion on systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse pressure (PP), and left ventricular hypertrophy (LVH) in two equal, sex- and age-matched groups; Grp1 and Grp2 (n = 100 each; age 53 ± 4 years) of hypertensive patients. Grp1 had regular GT treatment, followed by HW ingestion, whereas Grp2 had HW ingestion followed by GT treatment for periods of 4 months each. Electrocardiographic (ECG) and echocardiographic assessments of LVH were made before and at the end of both periods. SBP was lowered significantly by 6.6%; DBP by 5.1%, and PP by 9.1% by the end of month 4 of GT treatment in Grp1. Upon GT cessation and HW ingestion, SBP, DBP, and PP returned to pretreatment levels over 4 months. In Grp2, SBP, DBP, and PP were reduced insignificantly by 1.5%, 1.0%, and 2.3% by the end of the 4th month of HW ingestion. Conversely, over 4 months of GT treatment, SBP, DBP, and PP were significantly lowered by 5.4%, 4.1%, and 7.7% from the baseline values, respectively. ECG and echocardiographic evidence of LVH was shown in 20% of Grp1 and 24% of Grp2 patients before intervention. This was significantly lowered to 8% and 10% in Grp1 and Grp2 by GT treatment. However, this increased to 16% following HW ingestion in Grp1. HW ingestion did mot induce regression of LVH in Grp2. Thus, regular GT ingestion has cardiovascular protective effects.


Assuntos
Pressão Sanguínea , Hipertensão/dietoterapia , Hipertrofia Ventricular Esquerda/dietoterapia , Chá , Função Ventricular Esquerda , Remodelação Ventricular , Egito , Feminino , Humanos , Hipertensão/complicações , Hipertensão/diagnóstico , Hipertensão/fisiopatologia , Hipertrofia Ventricular Esquerda/diagnóstico por imagem , Hipertrofia Ventricular Esquerda/etiologia , Hipertrofia Ventricular Esquerda/fisiopatologia , Masculino , Pessoa de Meia-Idade , Fatores de Tempo , Resultado do Tratamento
6.
Nat Protoc ; 13(10): 2200-2216, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30218102

RESUMO

The goal of mechanobiology is to understand the links between changes in the physical properties of living cells and normal physiology and disease. This requires mechanical measurements that have appropriate spatial and temporal resolution within a single cell. Conventional atomic force microscopy (AFM) methods that acquire force curves pointwise are used to map the heterogeneous mechanical properties of cells. However, the resulting map acquisition time is much longer than that required to study many dynamic cellular processes. Dynamic AFM (dAFM) methods using resonant microcantilevers are compatible with higher-speed, high-resolution scanning; however, they do not directly acquire force curves and they require the conversion of a limited number of instrument observables to local mechanical property maps. We have recently developed a technique that allows commercial AFM systems equipped with direct cantilever excitation to quantitatively map the viscoelastic properties of live cells. The properties can be obtained at several widely spaced frequencies with nanometer-range spatial resolution and with fast image acquisition times (tens of seconds). Here, we describe detailed procedures for quantitative mapping, including sample preparation, AFM calibration, and data analysis. The protocol can be applied to different biological samples, including cells and viruses. The transition from dAFM imaging to quantitative mapping should be easily achievable for experienced AFM users, who will be able to set up the protocol in <30 min.


Assuntos
Biofísica/métodos , Microscopia de Força Atômica/métodos , Animais , Aplysia/citologia , Fenômenos Biomecânicos , Biofísica/instrumentação , Células Cultivadas , Elasticidade , Fibroblastos/citologia , Camundongos , Microscopia de Força Atômica/instrumentação , Células NIH 3T3 , Neurônios/citologia , Imagem Óptica , Viscosidade
7.
Clin Appl Thromb Hemost ; 24(6): 936-943, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28974109

RESUMO

T-cell immunoglobulin mucin 3 (TIM-3) is a transmembrane protein that plays an important role in several autoimmune diseases. The relationship between TIM-3 and excessive immune responses in immune thrombocytopenia (ITP) is still unknown. In this study, we evaluated the relationship between the expression of TIM-3 on peripheral blood mononuclear cells in patients with ITP and the disease severity. The frequency of lymphocyte and monocyte subsets and their TIM-3 expression were evaluated in patients with acute ITP (n = 45) and in healthy control (n = 20) using flow cytometry. Based on bleeding severity, patients were classified into 3 subgroups as mild (n = 12), moderate (n = 25), and severe (n = 8) bleeding. T-helper lymphocytes was found to be significantly decreased in the severe bleeding group compared to the mild and moderate bleeding groups, while CD56high natural killer (NK) cells were significantly expanded in severe bleeding group. In contrast, classical, intermediate, and nonclassical monocytes, natural killer T lymphocyte (NKT), and CD56dim NK cells showed no significant changes among different patient groups. This alteration of lymphocyte and monocyte subsets was associated with significant decrease in TIM-3 expression on CD56high NK cells, T-helper lymphocytes, NKT cells, and nonclassical monocytes in patients with ITP compared to the controls. Lower level of TIM-3 was found in severe bleeding group compared to mild and moderate bleeding groups. These results indicate that TIM-3 may be involved in the pathogenesis of ITP which subsequently can represent an opportunity for new therapeutic plan, moreover. This may have a prognostic value for disease severity.


Assuntos
Regulação da Expressão Gênica , Receptor Celular 2 do Vírus da Hepatite A/biossíntese , Monócitos/metabolismo , Células T Matadoras Naturais/metabolismo , Púrpura Trombocitopênica Idiopática/metabolismo , Linfócitos T Auxiliares-Indutores/metabolismo , Criança , Pré-Escolar , Feminino , Hemorragia/metabolismo , Hemorragia/patologia , Humanos , Masculino , Monócitos/patologia , Células T Matadoras Naturais/patologia , Púrpura Trombocitopênica Idiopática/patologia , Linfócitos T Auxiliares-Indutores/patologia
8.
Sci Rep ; 7(1): 7292, 2017 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-28779177

RESUMO

During the development of the nervous system and regeneration following injury, microtubules (MTs) are required for neurite elongation. Whether this elongation occurs primarily through tubulin assembly at the tip of the axon, the transport of individual MTs, or because MTs translocate forward in bulk is unclear. Using fluorescent speckle microscopy (FSM), differential interference contrast (DIC), and phase contrast microscopy, we tracked the movement of MTs, phase dense material, and docked mitochondria in chick sensory and Aplysia bag cell neurons growing rapidly on physiological substrates. In all cases, we find that MTs and other neuritic components move forward in bulk at a rate that on average matches the velocity of neurite elongation. To better understand whether and why MT assembly is required for bulk translocation, we disrupted it with nocodazole. We found this blocked the forward bulk advance of material along the neurite and was paired with a transient increase in axonal tension. This indicates that disruption of MT dynamics interferes with neurite outgrowth, not by disrupting the net assembly of MTs at the growth cone, but rather because it alters the balance of forces that power the bulk forward translocation of MTs.


Assuntos
Microtúbulos/metabolismo , Neuritos/metabolismo , Animais , Aplysia , Axônios/metabolismo , Biomarcadores , Microscopia de Fluorescência , Mitocôndrias/metabolismo , Transporte Proteico
9.
Rev. bras. entomol ; 61(2): 101-106, Apr.-Jun. 2017. tab, graf
Artigo em Inglês | LILACS | ID: biblio-843709

RESUMO

ABSTRACT During the present study, the host-parasite relationship between mosquitoes and parasitic mites was investigated. The 8954 individuals of male and female mosquitoes belonging to 26 genera: seven each of Aedes and Culex, six of Anopheles and one each of Toxorhynchites, Coquillettidia and Uranotaenia were collected from 200 sites. The male and female mosquitoes were collected from the State of Uttar Pradesh, located at 26.8500° N, 80.9100° E in North India by deploying Carbon dioxide-baited and gravid traps. The intensity of mite's infection, type and number of mites attached to mosquitoes, mite's preference for body parts and host sexes were the parameters used to determine host-parasite relationship. Eight species of mites: Arrenurus acuminatus, Ar. gibberifrons, Ar. danbyensis, Ar. madaraszi, Ar. kenki, Parathyas barbigera, Leptus sp., and Anystis sp., parasitized mosquitoes. Parasitic mites preferred host's thorax for attachment as compared to the head, pre-abdomen or appendages. The present study suggests phoretic relationship between parasitic mites and mosquitoes. Wide occurrence, intensity of infection, parasitic load, and attachment preferences of the mites suggested their positive role in biological control of adult mosquitoes. The present study will set the path of future studies on host-parasite relationships of mites and mosquitoes and define the role of parasitic mites in the biological control of mosquitoes.

10.
Arch Immunol Ther Exp (Warsz) ; 65(3): 263-269, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27638481

RESUMO

Gaucher disease (GD) is the most prevalent lysosomal storage disorder. Gaucher disease is associated with remarkable alterations in the immune system, and GD patients are more susceptible to infections and are at a higher risk of developing autoimmune disorders and malignancies. In a case-control study, we used three-color flow cytometric immunophenotyping for determination of the frequency of lymphocyte subpopulations and activated T lymphocytes among 18 children with GD1 under enzyme replacement therapy managed in Assiut University Hospitals. We found significant increases in the frequencies of total lymphocytes, CD19+, CD3+, CD4+, and CD8+ in children with GD1 when compared to healthy control. The frequencies of activated T lymphocytes (CD3+HLA-DR+), activated T-helper cells (CD4+HLA-DR+), and activated T-suppressor/cytotoxic cells (CD8+HLA-DR+) were significantly higher in GD1 as compared to healthy children. Our data show that the increased proportion of activated T lymphocytes in children with GD1 raises the issue of their possible involvement in the pathogenesis of the immune dysfunction seen in these patients. Our data suggested that the activated T lymphocytes could play a role in the clinical course of GD1. The relationship of these cells to immune disorders in GD1 children remains to be determined.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Doença de Gaucher/imunologia , Subpopulações de Linfócitos/imunologia , Antígenos CD/metabolismo , Estudos de Casos e Controles , Criança , Pré-Escolar , Terapia de Reposição de Enzimas , Feminino , Humanos , Memória Imunológica , Lactente , Ativação Linfocitária , Contagem de Linfócitos , Masculino
11.
Oxid Med Cell Longev ; 2016: 5296271, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26770655

RESUMO

Herbicides containing paraquat may contribute to the pathogenesis of neurodegenerative disorders such as Parkinson's disease. Paraquat induces reactive oxygen species-mediated apoptosis in neurons, which is a primary mechanism behind its toxicity. We sought to test the effectiveness of a commercially available polyphenol-rich Aronia melanocarpa (aronia berry) concentrate in the amelioration of paraquat-induced neurotoxicity. Considering the abundance of antioxidants in aronia berries, we hypothesized that aronia berry concentrate attenuates the paraquat-induced increase in reactive oxygen species and protects against paraquat-mediated neuronal cell death. Using a neuronal cell culture model, we observed that low doses of aronia berry concentrate protected against paraquat-mediated neurotoxicity. Additionally, low doses of the concentrate attenuated the paraquat-induced increase in superoxide, hydrogen peroxide, and oxidized glutathione levels. Interestingly, high doses of aronia berry concentrate increased neuronal superoxide levels independent of paraquat, while at the same time decreasing hydrogen peroxide. Moreover, high-dose aronia berry concentrate potentiated paraquat-induced superoxide production and neuronal cell death. In summary, aronia berry concentrate at low doses restores the homeostatic redox environment of neurons treated with paraquat, while high doses exacerbate the imbalance leading to further cell death. Our findings support that moderate levels of aronia berry concentrate may prevent reactive oxygen species-mediated neurotoxicity.


Assuntos
Neurotoxinas/toxicidade , Paraquat/toxicidade , Photinia/química , Extratos Vegetais/farmacologia , Antioxidantes/metabolismo , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Glutationa/metabolismo , Humanos , Peróxido de Hidrogênio/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/patologia , Oxidantes/toxicidade , Oxirredução/efeitos dos fármacos , Superóxidos/metabolismo
12.
Front Cell Neurosci ; 9: 359, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26441530

RESUMO

Mechanical force plays a fundamental role in neuronal development, physiology, and regeneration. In particular, research has shown that force is involved in growth cone-mediated axonal growth and guidance as well as stretch-induced elongation when an organism increases in size after forming initial synaptic connections. However, much of the details about the exact role of force in these fundamental processes remain unknown. In this review, we highlight: (1) standing questions concerning the role of mechanical force in axonal growth and guidance; and (2) different experimental techniques used to quantify forces in axons and growth cones. We believe that satisfying answers to these questions will require quantitative information about the relationship between elongation, forces, cytoskeletal dynamics, axonal transport, signaling, substrate adhesion, and stiffness contributing to directional growth advance. Furthermore, we address why a wide range of force values have been reported in the literature, and what these values mean in the context of neuronal mechanics. We hope that this review will provide a guide for those interested in studying the role of force in development and regeneration of neuronal networks.

13.
Biophys J ; 109(7): 1358-71, 2015 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-26445437

RESUMO

Although pulling forces have been observed in axonal growth for several decades, their underlying mechanisms, absolute magnitudes, and exact roles are not well understood. In this study, using two different experimental approaches, we quantified retrograde traction force in Aplysia californica neuronal growth cones as they develop over time in response to a new adhesion substrate. In the first approach, we developed a novel method, to our knowledge, for measuring traction forces using an atomic force microscope (AFM) with a cantilever that was modified with an Aplysia cell adhesion molecule (apCAM)-coated microbead. In the second approach, we used force-calibrated glass microneedles coated with apCAM ligands to guide growth cone advance. The traction force exerted by the growth cone was measured by monitoring the microneedle deflection using an optical microscope. Both approaches showed that Aplysia growth cones can develop traction forces in the 10(0)-10(2) nN range during adhesion-mediated advance. Moreover, our results suggest that the level of traction force is directly correlated to the stiffness of the microneedle, which is consistent with a reinforcement mechanism previously observed in other cell types. Interestingly, the absolute level of traction force did not correlate with growth cone advance toward the adhesion site, but the amount of microneedle deflection did. In cases of adhesion-mediated growth cone advance, the mean needle deflection was 1.05 ± 0.07 µm. By contrast, the mean deflection was significantly lower (0.48 ± 0.06 µm) when the growth cones did not advance. Our data support a hypothesis that adhesion complexes, which can undergo micron-scale elastic deformation, regulate the coupling between the retrogradely flowing actin cytoskeleton and apCAM substrates, stimulating growth cone advance if sufficiently abundant.


Assuntos
Adesão Celular/fisiologia , Movimento Celular/fisiologia , Elasticidade , Cones de Crescimento/fisiologia , Neurônios/fisiologia , Actinas/metabolismo , Animais , Aplysia , Moléculas de Adesão Celular/química , Células Cultivadas , Vidro , Microscopia de Força Atômica , Microesferas , Imagem Óptica
14.
J Bacteriol ; 196(23): 3983-91, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25157078

RESUMO

Raman spectroscopy was used to study the time course of phenotypic responses of Escherichia coli (DH5α) to 1-butanol exposure (1.2% [vol/vol]). Raman spectroscopy is of interest for bacterial phenotyping because it can be performed (i) in near real time, (ii) with minimal sample preparation (label-free), and (iii) with minimal spectral interference from water. Traditional off-line analytical methodologies were applied to both 1-butanol-treated and control cells to draw correlations with Raman data. Here, distinct sets of Raman bands are presented that characterize phenotypic traits of E. coli with maximized correlation to off-line measurements. In addition, the observed time course phenotypic responses of E. coli to 1.2% (vol/vol) 1-butanol exposure included the following: (i) decreased saturated fatty acids levels, (ii) retention of unsaturated fatty acids and low levels of cyclopropane fatty acids, (iii) increased membrane fluidity following the initial response of increased rigidity, and (iv) no changes in total protein content or protein-derived amino acid composition. For most phenotypic traits, correlation coefficients between Raman spectroscopy and traditional off-line analytical approaches exceeded 0.75, and major trends were captured. The results suggest that near-real-time Raman spectroscopy is suitable for approximating metabolic and physiological phenotyping of bacterial cells subjected to toxic environmental conditions.


Assuntos
1-Butanol/metabolismo , Escherichia coli/química , Escherichia coli/efeitos dos fármacos , Análise Espectral Raman , Proteínas de Bactérias/análise , Membrana Celular/química , Citosol/química , Ácidos Graxos/análise , Fatores de Tempo
15.
Blood Press ; 23(3): 160-7, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24059637

RESUMO

BACKGROUND AND AIM: Effects of Ramadan fasting on health are important. Its effects on arterial pulse pressure (PP), lipid profile and oxidative stress were characterized in hypertensives. METHODS: PP, indices of lipid profile and oxidative stress were measured pre-, during and post-fasting in equal (40 each), sex- and age-matched groups (age 55 ± 5 years) of hypertensives (HT) and controls (C). RESULTS: Fasting reduced PP significantly by 17.2% and insignificantly by 9.3% in the HT and C groups, respectively. Total cholesterol (TC) was lowered insignificantly by 11.7% and 4.7% in the HT and C patients, respectively. Triglycerides (TG) and malondialdehyde (MDA) were significantly lowered by: TG: 24.5% and 22.8%; MDA: 45.6% and 54.3%; while glutathione (GSH) elevated by 56.8% and 52.6% in the HT and C groups, respectively. High-density lipoproteins (HDL) were raised significantly by 33.3% and insignificantly by 6.7%, whereas low-density lipoproteins (LDL) decreased significantly by 17.7% and insignificantly by 4.0% in the HT and C groups, respectively. At 6 weeks post-fasting, MDA remained significantly lower than the pre-fasting level by 24.3% and 25.7%, and GSH higher by 30.2% and 26.3% in the HT and C groups, respectively, while PP and TC returned to pre-fasting values in both groups. The post-fasting, HDL was significantly higher by 20.3% and LDL lower by 12.0% than the fasting levels in the HT patients. CONCLUSION: Fasting improves PP and lipids profile and ameliorates oxidative stress in hypertensives.


Assuntos
Pressão Sanguínea/fisiologia , Jejum/fisiologia , Férias e Feriados , Hipertensão/prevenção & controle , Islamismo , Estresse Oxidativo/fisiologia , Colesterol/sangue , Jejum/sangue , Feminino , Glutationa/sangue , Humanos , Hipertensão/sangue , Lipoproteínas HDL/sangue , Lipoproteínas LDL/sangue , Masculino , Malondialdeído/sangue , Pessoa de Meia-Idade , Triglicerídeos/sangue
16.
Biotechnol J ; 8(5): 581-94, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23460591

RESUMO

Optimized production of bio-based fuels and chemicals from microbial cell factories is a central goal of systems metabolic engineering. To achieve this goal, a new computational method of using flux balance analysis with flux ratios (FBrAtio) was further developed in this research and applied to five case studies to evaluate and design metabolic engineering strategies. The approach was implemented using publicly available genome-scale metabolic flux models. Synthetic pathways were added to these models along with flux ratio constraints by FBrAtio to achieve increased (i) cellulose production from Arabidopsis thaliana; (ii) isobutanol production from Saccharomyces cerevisiae; (iii) acetone production from Synechocystis sp. PCC6803; (iv) H2 production from Escherichia coli MG1655; and (v) isopropanol, butanol, and ethanol (IBE) production from engineered Clostridium acetobutylicum. The FBrAtio approach was applied to each case to simulate a metabolic engineering strategy already implemented experimentally, and flux ratios were continually adjusted to find (i) the end-limit of increased production using the existing strategy, (ii) new potential strategies to increase production, and (iii) the impact of these metabolic engineering strategies on product yield and culture growth. The FBrAtio approach has the potential to design "fine-tuned" metabolic engineering strategies in silico that can be implemented directly with available genomic tools.


Assuntos
Biotecnologia/métodos , Engenharia Metabólica/métodos , Biologia de Sistemas/métodos , Álcoois/análise , Álcoois/metabolismo , Bactérias/genética , Bactérias/metabolismo , Biocombustíveis , Simulação por Computador , Genoma Bacteriano , Genoma Fúngico , Glucose/metabolismo , Microbiologia Industrial , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo
17.
Appl Environ Microbiol ; 78(21): 7805-8, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22923413

RESUMO

The ability to control the localization of surface-enhanced Raman scattering (SERS) nanoparticle probes in bacterial cells is critical to the development of analytical techniques that can nondestructively determine cell composition and phenotype. Here, selective localization of SERS probes was achieved at the outer bacterial membrane by using silver nanoparticles functionalized with synthetic hydrophobic peptides.


Assuntos
Proteínas da Membrana Bacteriana Externa/análise , Análise Espectral Raman/métodos , Bactérias , Estruturas Celulares , Nanopartículas Metálicas/química , Peptídeos , Fenótipo , Prata/química , Propriedades de Superfície
18.
J Magn Reson Imaging ; 22(5): 614-27, 2005 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16193471

RESUMO

PURPOSE: To measure cardiac blood flow patterns and ventricular wall velocities through the cardiac cycle in anesthetized Wistar Kyoto (WKY) rats. MATERIALS AND METHODS: A gradient-echo cine pulse sequence incorporating pulsed field gradients (PFGs) provided phase contrast (PC) motion encoding. We achieved a range of velocity sensitivity that was sufficient to measure simultaneously the large flow velocities within the cardiac chambers and aortic outflow tract (up to 70 cm s(-1) during systole), and the comparatively small velocities of the cardiac wall (0-3 cm s(-1)). A scheme of sparsely sampling q-space combined with a probability-based method of velocity calculation permitted such measurements along three orthogonal axes, and yielded velocity vector maps in all four chambers of the heart and the aorta, in both longitudinal and transverse sections, for up to 12 time-points in the cardiac cycle. RESULTS: Left ventricular systole was associated with a symmetrical laminar flow pattern along the cardiac axis, with no appearance of turbulence. In contrast, blood showed a swirling motion within the right ventricle (RV) in the region of the pulmonary outflow tract. During left ventricular diastole a plume of blood entered the left ventricle (LV) from the left atrium. The ventricular flow patterns could also be correlated with measurements of left ventricular wall motion. The greatest velocities of the ventricular walls occurred in the transverse cardiac plane and were maximal during diastolic refilling. The cardiac wall motion in the longitudinal axis demonstrated a caudal-apical movement that may also contribute to diastolic refilling. CONCLUSION: The successful measurements of blood and myocardial velocity during normal myocardial function may be extended to quantify pathological cardiac changes in animal models of human cardiac disease.


Assuntos
Circulação Coronária/fisiologia , Aumento da Imagem/métodos , Interpretação de Imagem Assistida por Computador/métodos , Imageamento por Ressonância Magnética/métodos , Contração Miocárdica/fisiologia , Processamento de Sinais Assistido por Computador , Função Ventricular Esquerda/fisiologia , Algoritmos , Animais , Velocidade do Fluxo Sanguíneo/fisiologia , Ventrículos do Coração/anatomia & histologia , Movimento/fisiologia , Ratos , Ratos Endogâmicos WKY , Função Ventricular
19.
J Physiol ; 538(Pt 2): 541-53, 2002 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-11790818

RESUMO

A non-invasive cine magnetic resonance imaging (MRI) technique was developed to allow, for the first time, detection and characterization of chronic changes in myocardial tissue volume and the effects upon these of treatment by the angiotensin-converting enzyme (ACE) inhibitor captopril in streptozotocin (STZ)-diabetic male Wistar rats. Animals that had been made diabetic at the ages of 7, 10 and 13 weeks and a captopril-treated group of animals made diabetic at the age of 7 weeks were scanned. The findings were compared with the results from age-matched controls. All animal groups (n = 4 animals in each) were consistently scanned at 16 weeks. Left and right ventricular myocardial volumes were reconstructed from complete data sets of left and right ventricular transverse sections which covered systole and most of diastole using twelve equally incremented time points through the cardiac cycle. The calculated volumes remained consistent through all twelve time points of the cardiac cycle in all five experimental groups and agreed with the corresponding post-mortem determinations. These gave consistent myocardial densities whose values could additionally be corroborated by previous reports, confirming the validity of the quantitative MRI results and analysis. The myocardial volumes were conserved in animals whose diabetes was induced at 13 weeks but were significantly increased relative to body weight in animals made diabetic at 7 and 10 weeks. Captopril treatment, which was started immediately after induction of diabetes, prevented the development of this relative hypertrophy in both the left and right ventricles. We have thus introduced and validated quantitative MRI methods in a demonstration, for the first time, of chronic myocardial changes in both the right and left ventricles of STZ-diabetic rats and their prevention by the ACE inhibitor captopril.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Captopril/farmacologia , Diabetes Mellitus Experimental/patologia , Imagem Cinética por Ressonância Magnética , Miocárdio/patologia , Animais , Diabetes Mellitus Experimental/fisiopatologia , Coração/efeitos dos fármacos , Coração/fisiopatologia , Hipertrofia Ventricular Esquerda/prevenção & controle , Hipertrofia Ventricular Direita/prevenção & controle , Masculino , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Wistar
20.
J Physiol ; 538(Pt 2): 555-72, 2002 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-11790819

RESUMO

Non-invasive magnetic resonance imaging (MRI) was used to characterize changes in left and right ventricular cardiac cycles following induction of experimental, streptozotocin (STZ)-induced, diabetes in male Wistar rats at different ages. The effects of the angiotensin-converting enzyme (ACE) inhibitor captopril upon such chronic physiological changes were then evaluated, also for the first time. Diabetes was induced at the age of 7 weeks in two experimental groups, of which one group was subsequently maintained on captopril (2 g l(-1))-containing drinking water, and at 10 and 13 weeks in two further groups. The fifth group provided age-matched controls. All groups (each n = 4 animals) were scanned consistently at 16 weeks, in parallel with timings used in earlier studies that employed this experimental model. Cine magnetic resonance (MR) image acquisition provided transverse sections through both ventricles at twelve time points covering systole and most of diastole. These yielded reconstructions of cardiac anatomy used to derive critical functional indices and their dependence upon time following the triggering electrocardiographic R waves. The left and right ventricular end-diastolic (EDV), end-systolic (ESV) and stroke volumes (SV), and ejection fractions (EF) calculated from each, control and experimental, group showed matching values. This confirmed a necessary condition requiring balanced right and left ventricular outputs and further suggested that STZ-induced diabetes produced physiological changes in both ventricles. Absolute left and right ventricular SVs were significantly altered in all diabetic animals; EDVs and EFs significantly altered in animals diabetic from 7 and 10 but not 13 weeks. When normalized to body weight, left and right ventricular SVs had significantly altered in animals diabetic from 7 and 10 weeks but not 13 weeks. Normalized left ventricular EDVs were also significantly altered in animals diabetic from 7 and 10 weeks. However, normalized right ventricular EDVs were significantly altered only in animals made diabetic from 7 weeks. Diabetic hearts showed major kinetic changes in left and right ventricular contraction (ejection) and relaxation (filling). Both the initial rates of volume change (dV/dt) in both ventricles and the plots of dV/dt values through the cardiac cycle demonstrated more gradual developments of tension during systole and relaxation during diastole. Estimates of the derived left ventricular performance parameters of cardiac output, cardiac power output and stroke work in control animals were comparable with human values when normalized to both body (or cardiac) weight and heart rate. All deteriorated with diabetes. Comparisons of experimental groups diabetic from 7 weeks demonstrated that captopril treatment relieved the alterations in critical volumes, dependence of SV upon EDV, kinetics of systolic contraction and diastolic relaxation and in the derived indicators of ventricular performance. This study represents the first demonstration using non-invasive MRI of early, chronic changes in diastolic filling and systolic ejection in both the left and the right ventricles and of their amelioration by ACE inhibition following STZ-induction of diabetes in intact experimental animals.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Captopril/farmacologia , Diabetes Mellitus Experimental/fisiopatologia , Coração/efeitos dos fármacos , Coração/fisiopatologia , Imagem Cinética por Ressonância Magnética , Animais , Circulação Coronária , Diabetes Mellitus Experimental/diagnóstico , Masculino , Contração Miocárdica , Ratos , Ratos Wistar , Volume Sistólico , Função Ventricular Esquerda/efeitos dos fármacos
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