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1.
Cell ; 183(6): 1562-1571.e12, 2020 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-33306955

RESUMO

Ticks transmit a diverse array of microbes to vertebrate hosts, including human pathogens, which has led to a human-centric focus in this vector system. Far less is known about pathogens of ticks themselves. Here, we discover that a toxin in blacklegged ticks (Ixodes scapularis) horizontally acquired from bacteria-called domesticated amidase effector 2 (dae2)-has evolved to kill mammalian skin microbes with remarkable efficiency. Secreted into the saliva and gut of ticks, Dae2 limits skin-associated staphylococci in ticks while feeding. In contrast, Dae2 has no intrinsic ability to kill Borrelia burgdorferi, the tick-borne Lyme disease bacterial pathogen. These findings suggest ticks resist their own pathogens while tolerating symbionts. Thus, just as tick symbionts can be pathogenic to humans, mammalian commensals can be harmful to ticks. Our study underscores how virulence is context-dependent and bolsters the idea that "pathogen" is a status and not an identity.


Assuntos
Bactérias/metabolismo , Fatores Imunológicos/metabolismo , Ixodes/fisiologia , Pele/microbiologia , Simbiose , Animais , Antibacterianos/farmacologia , Biocatálise , Parede Celular/metabolismo , Comportamento Alimentar , Feminino , Trato Gastrointestinal/metabolismo , Interações Hospedeiro-Patógeno , Camundongos , Modelos Moleculares , Peptidoglicano/metabolismo , Filogenia , Saliva/metabolismo , Glândulas Salivares/metabolismo , Staphylococcus epidermidis/fisiologia , Homologia Estrutural de Proteína , Especificidade por Substrato , Regulação para Cima
2.
Proc Natl Acad Sci U S A ; 120(25): e2209810120, 2023 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-37307472

RESUMO

Patients with type 1 diabetes (T1D) suffer from insufficient functional ß-cell mass, which results from infiltration of inflammatory cells and cytokine-mediated ß-cell death. Previous studies demonstrated the beneficial effects of agonists of growth hormone-releasing hormone receptor (GHRH-R), such as MR-409 on preconditioning of islets in a transplantation model. However, the therapeutic potential and protective mechanisms of GHRH-R agonists on models of T1D diabetes have not been explored. Using in vitro and in vivo models of T1D, we assessed the protective propertie of the GHRH agonist, MR409 on ß-cells. The treatment of insulinoma cell lines and rodent and human islets with MR-409 induces Akt signaling by induction of insulin receptor substrate 2 (IRS2), a master regulator of survival and growth in ß-cells, in a PKA-dependent manner. The increase in cAMP/PKA/CREB/IRS2 axis by MR409 was associated with decrease in ß-cell death and improved insulin secretory function in mouse and human islets exposed to proinflammatory cytokines. The assessment of the effects of GHRH agonist MR-409 in a model of T1D induced by low-dose streptozotocin showed that mice treated with MR-409 exhibited better glucose homeostasis, higher insulin levels, and preservation of ß-cell mass. Increased IRS2 expression in ß-cells in the group treated with MR-409 corroborated the in vitro data and provided evidence for the underlying mechanism responsible for beneficial effects of MR-409 in vivo. Collectively, our data show that MR-409 is a novel therapeutic agent for the prevention and treatment of ß-cells death in T1D.


Assuntos
Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 1 , Neoplasias Pancreáticas , Humanos , Animais , Camundongos , Estreptozocina , Citocinas , Insulina
3.
J Chem Phys ; 158(22)2023 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-37309896

RESUMO

The incorporation of quantum dots in display technology has fueled a renewed interest in InP-based quantum dots, but difficulty controlling the Zn chemistry during shelling has stymied thick, even ZnSe shell growth. The characteristic uneven, lobed morphology of Zn-based shells is difficult to assess qualitatively and measure through traditional methods. Here, we present a methodological study utilizing quantitative morphological analysis of InP/ZnSe quantum dots to analyze the impact of key shelling parameters on InP core passivation and shell epitaxy. We compare conventional hand-drawn measurements with an open-source semi-automated protocol to showcase the improved precision and speed of this method. Additionally, we find that quantitative morphological assessment can discern morphological trends in morphologies that qualitative methods cannot. In conjunction with ensemble fluorescence measurements, we find that changes to shelling parameters that promote even shell growth often do so at the cost of core homogeneity. These results indicate that the chemistry of passivating the core and promoting shell growth must be balanced carefully to maximize brightness while maintaining emission color-purity.

4.
Rev Argent Microbiol ; 53(2): 154-161, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33176955

RESUMO

St. Louis encephalitis (SLEV) and West Nile (WNV) arboviruses, which circulate in Argentina, are maintained in enzootic transmission cycles involving Culex mosquitoes (vectors) and birds belonging to orders Passeriformes and Columbiformes (amplifier hosts). The objective of this work was to determine the circulation of both viruses among wild birds in a semiarid ecosystem in the Province of La Rioja through a serologic survey. During spring 2013 and fall 2014, a total of 326 wild birds belonging to 41 species were captured in areas close to the cities of La Rioja and Chilecito, in the Province of La Rioja. While exposure to SLEV and WNV was analyzed in birds' serum through neutralizing antibody detection, viral circulation was estimated through apparent seroprevalence of neutralizing antibodies. The exposure of the avian community to viruses was 3.02% for SLEV and 1.89% for WNV, while 1.19% corresponded to coinfections. Our study confirms for the first time the circulation of SLEV and WNV in wild birds in the Province of La Rioja. Moreover, it is the first study to register neutralizing antibodies for flavivirus in the species Leptotila verreauxi (White-tipped Dove) (WNV) and Melanerpes cactorum (White-fronted Woodpecker) (SLEV). These results suggest that in semiarid ecosystems from northwestern Argentina the requirements and conditions for amplification and enzootic maintenance of SLEV and WNV would be present.


Assuntos
Encefalite de St. Louis , Vírus do Nilo Ocidental , Animais , Anticorpos Antivirais , Argentina/epidemiologia , Ecossistema , Vírus da Encefalite de St. Louis , Encefalite de St. Louis/epidemiologia , Encefalite de St. Louis/veterinária , Estudos Soroepidemiológicos
5.
Eur Spine J ; 27(10): 2577-2583, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29995170

RESUMO

PURPOSE: Differentiating osteoporotic vertebral fractures (OVF) from metastatic vertebral fractures (MVF) is difficult. A magnetic resonance imaging (MRI)-based score (META score) aiming to differentiate OVF and MVF was recently published; however, an independent agreement assessment is required before the score is used. We performed such independent agreement evaluation, including raters with different levels of training. METHODS: Sixty-four patients with confirmed OVF or MVF were evaluated by six raters (three spine surgeons and three orthopaedic residents) using the META score. We used the intra-class correlation coefficient (ICC) to evaluate inter- and intra-observer agreement and the kappa statistic (κ) to determine the agreement for individual score criteria. We calculated the area under the receiver-operating characteristic curve (AUC) to establish the score accuracy. RESULTS: The inter-observer agreement was poor [ICC = 0.22 (0.12-0.33)]; spine surgeons [ICC = 0.75 (0.66-0.83)] had better agreement than that of residents [ICC = 0.06 (- 0.07 to 0.23)]. The intra-observer agreement was poor [ICC = 0.15 (- 0.04 to 0.30)]; both spine surgeons [ICC = 0.21 (0.05-0.41)] and residents exhibited poor agreement [ICC = - 0.06 (- 0.40 to 0.20)]. The agreement for each specific criterion varied from κ = 0.24 to κ = 0.38. The AUC was 0.57 (0.64 for spine surgeons and 0.51 for residents, p < 0.01). CONCLUSION: The inter-observer agreement using the META score was adequate for spine surgeons but not for residents; the intra-observer agreement was poor. These results do not support the standard use of the META score to differentiate OVF and MVF. These slides can be retrieved under Electronic Supplementary Material.


Assuntos
Diagnóstico por Computador/métodos , Imageamento por Ressonância Magnética , Fraturas por Osteoporose/diagnóstico por imagem , Fraturas da Coluna Vertebral/diagnóstico por imagem , Diagnóstico Diferencial , Humanos , Fraturas por Osteoporose/cirurgia , Curva ROC , Fraturas da Coluna Vertebral/cirurgia
6.
J Theor Biol ; 418: 122-128, 2017 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-28130095

RESUMO

In 1974, Eisenthal and Cornish-Bowden published the direct linear plot method, which used the median to estimate the Vmax and Km from a set of initial rates as a function of substrate concentrations. The robustness of this non-parametric method was clearly demonstrated by comparing it with the least-squares method. The authors commented that the method cannot readily be generalized to equations of more than two parameters. Unfortunately, this comment has been misread by other authors. Comments such as "this method cannot be extended directly to equations with more than two parameters" were found in some publications. In addition, recently, the most drastic comment was published: "this method cannot be applied for the analysis of substrate inhibition." Given all of these presumptions, we have been motivated to publish a demonstration of the contrary: the median method can be applied to more than two-parameter equations, using as an example, the substrate uncompetitive inhibition equation. A computer algorithm was written to evaluate the effect of simulated experimental error of the initial rates on the estimation of Vmax, Km and KS. The error was assigned to different points of the experimental design. Four different KS/Km ratios were analyzed with the values 10, 100, 1000 and 10,000. The results indicated that the least-squares method was slightly better than the median method in terms of accuracy and variance. However, the presence of outliers affected the estimation of kinetic constants using the least-squares method more severely than the median method. The estimation of KS using the median method to estimate 1/KS was much better than the direct estimation of KS, causing a negative effect of non-linearity of KS in the kinetic equation. Considering that the median method is free from the assumptions of the least-squares method and the arbitrary assumptions implicit in the linearization methods to estimate the kinetic constants Vmax, Km and KS from the substrate uncompetitive inhibition equation, the median method is highly superior to all published methods, including the non-linear regression by least squares. We concluded that the median method can be applied to the substrate uncompetitive inhibition equation and other equations with more than two parameters. In addition, as we can project, the median method is the most reliable and robust method for the estimation of kinetic parameters from enzyme kinetic models.


Assuntos
Modelos Químicos , Cinética
7.
Biometals ; 28(1): 143-50, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25428685

RESUMO

Despite the evidence suggesting a role for Helicobacter pylori in the induction of systemic iron deficiency anaemia, little is known about the possibility of infection-associated changes in cellular iron homeostasis at the gastric epithelium. In this study we compared four different techniques for measuring iron in AGS cells, a gastric epithelial cell line that is widely used to model to H. pylori infection in vitro. Inductively coupled plasma-mass spectrometry proved to be an efficient method, but only when large numbers of cells were used. Two colorimetric assays that included the use of concentrated hydrochloric acid with or without potassium ferrocyanide detected iron in the micromolar but not the nanomolar range in cell-free standards. However, the third colorimetric assay that incorporated ferrozine proved to be highly accurate at detecting iron in the nanomolar range, and was able to detect iron in AGS cells, Moreover, using this assay, we were able to show that the level of iron in H. pylori-infected AGS cells is significantly increased when compared to uninfected cells.


Assuntos
Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Mucosa Gástrica/citologia , Helicobacter pylori/patogenicidade , Ferro/metabolismo , Linhagem Celular Tumoral , Infecções por Helicobacter/fisiopatologia , Humanos
8.
Biometals ; 27(1): 115-23, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24327293

RESUMO

Divalent Metal Transporter 1 (DMT1) is an apical Fe transporter in the duodenum and is involved in endosomal Fe export. Four protein isoforms have been described for DMT1, two from mRNA with an iron responsive element (IRE) and two from mRNA without it. The sets of two begin in exon 1A or 2. We have characterized copper transport using mouse 2/-IRE DMT1 during regulated ectopic expression. HEK293 cells carrying a TetR:Hyg element were stably transfected with pDEST31 containing a 2/-IRE construct. (64)Cu(1+) incorporation in doxycycline treated cells exhibited 18.6 and 30.0-fold increases in Cu content, respectively when were exposed to 10 and 100 µM of extracellular Cu. Cu content was ~4-fold above that of parent cells or cells carrying just the vector. (64)Cu uptake in transfected cells pre-incubated with 5 µM of Cu-His revealed a Vmax and Km of 11.98 ± 0.52 pmol mg protein(-1) min(-1) and 2.03 ± 0.03 µM, respectively. Doxycycline-stimulated Cu uptake was linear with time. The rates of apical Cu uptake decreased and transepithelial transport increased when intracellular Cu increased. The optimal pH for Cu transport was 6.5; uptake of Cu was temperature dependent. Silver does not inhibit Cu uptake in cells carrying the vector. In conclusion, Cu uptake in HEK293 cells that over-expressed the 2/-IRE isoform of DMT1 transporter supports our earlier contention that DMT1 transports Cu as Cu(1+).


Assuntos
Proteínas de Transporte de Cátions/metabolismo , Cobre/metabolismo , Animais , Transporte Biológico , Células HEK293 , Humanos , Camundongos
9.
Elife ; 112022 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-35762582

RESUMO

Members of the bacterial T6SS amidase effector (Tae) superfamily of toxins are delivered between competing bacteria to degrade cell wall peptidoglycan. Although Taes share a common substrate, they exhibit distinct antimicrobial potency across different competitor species. To investigate the molecular basis governing these differences, we quantitatively defined the functional determinants of Tae1 from Pseudomonas aeruginosa PAO1 using a combination of nuclear magnetic resonance and a high-throughput in vivo genetic approach called deep mutational scanning (DMS). As expected, combined analyses confirmed the role of critical residues near the Tae1 catalytic center. Unexpectedly, DMS revealed substantial contributions to enzymatic activity from a much larger, ring-like functional hot spot extending around the entire circumference of the enzyme. Comparative DMS across distinct growth conditions highlighted how functional contribution of different surfaces is highly context-dependent, varying alongside composition of targeted cell walls. These observations suggest that Tae1 engages with the intact cell wall network through a more distributed three-dimensional interaction interface than previously appreciated, providing an explanation for observed differences in antimicrobial potency across divergent Gram-negative competitors. Further binding studies of several Tae1 variants with their cognate immunity protein demonstrate that requirements to maintain protection from Tae activity may be a significant constraint on the mutational landscape of tae1 toxicity in the wild. In total, our work reveals that Tae diversification has likely been shaped by multiple independent pressures to maintain interactions with binding partners that vary across bacterial species and conditions.


Assuntos
Amidoidrolases , Peptidoglicano , Amidoidrolases/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Parede Celular/metabolismo , Peptidoglicano/metabolismo , Pseudomonas aeruginosa/metabolismo
10.
J Nutr ; 141(9): 1652-6, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21795430

RESUMO

Calcium is the only known component in the diet that may affect absorption of both nonheme and heme iron. However, the evidence for a calcium effect on iron absorption mainly comes from studies that did not isolate the effect of calcium from that of other dietary components, because it was detected in single-meal studies. Our objective was to establish potential effects of calcium on absorption of nonheme and heme iron and the dose response for this effect in the absence of a meal. Fifty-four healthy, nonpregnant women were selected to participate in 4 iron absorption studies using iron radioactive tracers. We evaluated the effects of calcium doses between 200 and 1500 mg on absorption of 5 mg nonheme iron (as ferrous sulfate). We also evaluated the effects of calcium doses between 200 and 800 mg on absorption of 5 mg heme iron [as concentrated RBC (CRBC)]. Calcium was administered as calcium chloride in all studies and minerals were ingested on an empty stomach. Calcium doses ≥1000 mg diminished nonheme iron absorption by an average of 49.6%. A calcium dose of 800 mg diminished absorption of 5 mg heme iron by 37.7%. In conclusion, we demonstrated an isolated effect of calcium (as chloride) on absorption of 5 mg of iron provided as nonheme (as sulfate) and heme (as CRBC) iron. This effect was observed at doses higher than previously reported from single-meal studies, starting at ~800 mg of calcium.


Assuntos
Cálcio/farmacologia , Heme/farmacocinética , Ferro/administração & dosagem , Ferro/farmacocinética , Adulto , Anemia Ferropriva/tratamento farmacológico , Cálcio/administração & dosagem , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Interações Medicamentosas , Feminino , Humanos , Ferro/uso terapêutico , Pessoa de Meia-Idade
11.
Eur J Nutr ; 50(8): 637-43, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21327555

RESUMO

OBJECTIVE: To study heme iron bioavailability and the role of dietary protein (animal and vegetable) on iron uptake using an in vitro model (Caco-2 cell line). METHODS: Caco-2 cells were seeded in bicameral chambers with different animal (beef, chicken or fish) or vegetable (peas, lentils, and soybeans) proteins or with pure animal (collagen and casein) or vegetable (gliadin, zein, and glutein) protein extracts. The effect of each protein over heme iron absorption was assessed. RESULTS: Intact heme uptake was higher than either heme plus albumin or digested heme plus albumin, but lower than digested heme. White meal exerted the highest inhibitory effect on hemin uptake. Heme iron uptake decreased in the presence of all legume extracts, but was not significantly different among them (one-way ANOVA, NS). Pure animal (collagen and casein) and vegetable (zein and glutelin) proteins increased heme iron uptake, except for gliadin. CONCLUSION: Animal and vegetable protein in general decreased heme iron uptake. However, purified animal and vegetable protein induce an increase in heme iron uptake.


Assuntos
Proteínas Alimentares/administração & dosagem , Heme/farmacocinética , Ferro da Dieta/farmacocinética , Carne , Proteínas de Vegetais Comestíveis/administração & dosagem , Absorção , Animais , Disponibilidade Biológica , Células CACO-2 , Caseínas/metabolismo , Bovinos , Galinhas , Colágeno/metabolismo , Fabaceae , Peixes , Glutens/metabolismo , Humanos , Zeína/metabolismo
12.
J Vis Exp ; (145)2019 03 07.
Artigo em Inglês | MEDLINE | ID: mdl-30907885

RESUMO

The Solute Carrier 4 (SLC4) family of proteins is called the bicarbonate transporters and includes the archetypal protein Anion Exchanger 1 (AE1, also known as Band 3), the most abundant membrane protein in the red blood cells. The SLC4 family is homologous with borate transporters, which have been characterized in plants and fungi. It remains a significant technical challenge to express and purify membrane transport proteins to homogeneity in quantities suitable for structural or functional studies. Here we describe detailed procedures for the overexpression of borate transporters in Saccharomyces cerevisiae, isolation of yeast membranes, solubilization of protein by detergent, and purification of borate transporter homologs from S. cerevisiae, Arabidopsis thaliana, and Oryza sativa. We also detail a glutaraldehyde cross-linking experiment to assay multimerization of homomeric transporters. Our generalized procedures can be applied to all three proteins and have been optimized for efficacy. Many of the strategies developed here can be utilized for the study of other challenging membrane proteins.


Assuntos
Boratos/metabolismo , Células Eucarióticas/metabolismo , Proteínas de Membrana Transportadoras/isolamento & purificação , Proteínas de Membrana Transportadoras/metabolismo , Arabidopsis/metabolismo , Cromatografia em Gel , Reagentes de Ligações Cruzadas/metabolismo , Transporte de Íons , Oryza/metabolismo , Saccharomyces cerevisiae/metabolismo , Solubilidade
13.
Biol Trace Elem Res ; 125(2): 109-19, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18528644

RESUMO

It is known that heme iron and inorganic iron are absorbed differently. Heme iron is found in the diet mainly in the form of hemoglobin and myoglobin. The mechanism of iron absorption remains uncertain. This study focused on the heme iron uptake by Caco-2 cells from a hemoglobin digest and its response to different iron concentrations. We studied the intracellular Fe concentration and the effect of time, K+ depletion, and cytosol acidification on apical uptake and transepithelial transport in cells incubated with different heme Fe concentrations. Cells incubated with hemoglobin-digest showed a lower intracellular Fe concentration than cells grown with inorganic Fe. However, uptake and transepithelial transport of Fe was higher in cells incubated with heme Fe. Heme Fe uptake had a low Vmax and Km as compared to inorganic Fe uptake and did not compete with non-heme Fe uptake. Heme Fe uptake was inhibited in cells exposed to K+ depletion or cytosol acidification. Heme oxygenase 1 expression increased and DMT1 expression decreased with higher heme Fe concentrations in the media. The uptake of heme iron is a saturable and temperature-dependent process and, therefore, could occur through a mechanism involving both a receptor and the endocytic pathway.


Assuntos
Heme/metabolismo , Ferro/metabolismo , Potássio/farmacologia , Temperatura , Células CACO-2 , Proteínas de Transporte de Cátions/metabolismo , Endocitose/efeitos dos fármacos , Heme Oxigenase (Desciclizante)/metabolismo , Heme Oxigenase-1/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Cinética
14.
Biol Trace Elem Res ; 181(1): 38-43, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28500577

RESUMO

Pectins are a type of soluble fiber present in natural and processed foods. Evidence regarding the effect of esterification degree of pectins on iron absorption in humans is scarce. In the present study, the effect of pectins with different degrees of esterification on non-heme iron absorption in women was evaluated. A controlled experimental study was conducted with block design, involving 13 apparently healthy, adult women. Each subject received 5 mg Fe (FeSO4) without pectin (control) or accompanied by 5 g citrus pectin, two with a low degree of esterification (27 and 36%), and one with a high degree of esterification (67 to 73%), each on different days. Each day, the 5 mg Fe doses were marked with radioactive 59Fe or 55Fe. Radioactivity incorporated into erythrocytes was determined in blood samples 14 days after the marked Fe doses were consumed. On days 18 and 36 of study, 30 and 20 mL blood samples were obtained, respectively, and blood sample radioactivity incorporated into erythrocytes was determined. Body iron status was determined from blood taken on day 18. Whole body blood volume was estimated for calculate iron bioavailability; it was assumed that 80% of absorbed radioactivity was incorporated into the Hb. All women participants signed an informed consent of participation at baseline. Iron bioavailability (mean geometric ±1 SD) alone (control) was 18.2% (12.3-27.1%), iron + pectin27 was 17.2% (10.2-29.2%), iron + pectin36 was 15.3% (9.5-24.6%), and iron + pectin67 was 19.5% (10.0-38.0%). No statistically significant differences between iron bioavailability (repeated measures ANOVA, p = 0.22) were observed. Pectin esterification degree does not influence the bioavailability of non-heme iron in women.


Assuntos
Ésteres/metabolismo , Ferro/metabolismo , Pectinas/metabolismo , Adulto , Disponibilidade Biológica , Ésteres/sangue , Feminino , Humanos , Ferro/administração & dosagem , Ferro/sangue , Pessoa de Meia-Idade , Pectinas/administração & dosagem , Pectinas/sangue , Software
15.
Eur J Med Chem ; 146: 621-635, 2018 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-29407986

RESUMO

The present study seeks to describe the design and synthesis of six new Michael adducts of (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-dithiocarbamate with nitrostyrenes and their in vitro antiproliferative activity against human cervical cancer cell lines [HeLa (HPV 18 positive), CaSki (HPV 16 positive) and ViBo (HPV negative) cervical cancer cell lines]. Virtual screening of the physicochemical properties of all compounds have also been presented. All the compounds exploited significant antiproliferative activity on the three cervical cancer cell lines. Compound 8a was found to be most potent, displaying in vitro antiproliferative activity against HeLa, CaSki and ViBo cervical cancer cell lines superior to Cisplatin and Paclitaxel with IC50 values 0.99 ±â€¯0.007, 2.36 ±â€¯0.016 and 0.73 ±â€¯0.002 µM respectively. In addition, compound 8a did not trigger the necrosis cell death to the test cancer cell lines. Further mechanistic study revealed that compound 8a could inhibit the cancer cell proliferation by inducing apoptosis through caspase-3 activation. Moreover, cell cycle analysis indicated that compound 8a could arrest the cell cycle at the G1 phase for HeLa and CaSki cancer cells. At the predetermined IC50 values on cancer cells, compound 8a did not induce any necrotic (cytotoxic) death to the normal human lymphocytes. In the present design, (1S,4S)-2,5-diazabicyclo[2.2.1]heptane system was found to be superior than the piperazine counterpart 11.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Compostos Aza/farmacologia , Nitrocompostos/farmacologia , Estireno/farmacologia , Tiocarbamatos/farmacologia , Neoplasias do Colo do Útero/tratamento farmacológico , Antineoplásicos/síntese química , Antineoplásicos/química , Compostos Aza/química , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Células HeLa , Humanos , Estrutura Molecular , Nitrocompostos/química , Relação Estrutura-Atividade , Estireno/química , Tiocarbamatos/química , Células Tumorais Cultivadas , Neoplasias do Colo do Útero/patologia
16.
PLoS One ; 12(9): e0184026, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28873091

RESUMO

The iron deficiency anaemia that often accompanies infection with Helicobacter pylori may reflect increased uptake of iron into gastric epithelial cells. Here we show an infection-associated increase in total intracellular iron levels was associated with the redistribution of the transferrin receptor from the cell cytosol to the cell surface, and with increased levels of ferritin, an intracellular iron storage protein that corresponded with a significant increase in lysosomal stores of labile iron. In contrast, the pool of cytosolic labile iron was significantly decreased in infected cells. These changes in intracellular iron distribution were associated with the uptake and trafficking of H. pylori through the cells, and enhanced in strains capable of expressing the cagA virulence gene. We speculate that degradation of lysosomal ferritin may facilitate H. pylori pathogenesis, in addition to contributing to bacterial persistence in the human stomach.


Assuntos
Células Epiteliais/metabolismo , Células Epiteliais/patologia , Infecções por Helicobacter/microbiologia , Infecções por Helicobacter/patologia , Helicobacter pylori/fisiologia , Homeostase , Ferro/metabolismo , Estômago/patologia , Antígenos de Bactérias/metabolismo , Proteínas de Bactérias/metabolismo , Compartimento Celular , Linhagem Celular Tumoral , Células Epiteliais/microbiologia , Ferritinas/metabolismo , Humanos , Lisossomos/metabolismo , Mutação/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores da Transferrina/genética , Receptores da Transferrina/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , proteínas de unión al GTP Rab7
17.
Data Brief ; 11: 567-571, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28349104

RESUMO

We provide initial rate data from enzymatic reaction experiments and tis processing to estimate the kinetic parameters from the substrate uncompetitive inhibition equation using the median method published by Eisenthal and Cornish-Bowden (Cornish-Bowden and Eisenthal, 1974; Eisenthal and Cornish-Bowden, 1974). The method was denominated the direct linear plot and consists in the calculation of the median from a dataset of kinetic parameters Vmax and Km from the Michaelis-Menten equation. In this opportunity we present the procedure to applicate the direct linear plot to the substrate uncompetitive inhibition equation; a three-parameter equation. The median method is characterized for its robustness and its insensibility to outlier. The calculations are presented in an Excel datasheet and a computational algorithm was developed in the free software Python. The kinetic parameters of the substrate uncompetitive inhibition equation Vmax , Km and Ks were calculated using three experimental points from the dataset formed by 13 experimental points. All the 286 combinations were calculated. The dataset of kinetic parameters resulting from this combinatorial was used to calculate the median which corresponds to the statistic estimator of the real kinetic parameters. A comparative statistical analyses between the median method and the least squares was published in Valencia et al. [3].

18.
Medwave ; 16 Suppl 1: e6383, 2016 Mar 15.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-27028069

RESUMO

Thoracolumbar burst fractures account for up to 17% of major spinal fractures. Both conservative and operative treatments are widely used in clinical practice to manage thoracolumbar burst fractures. Previous studies showed good functional results with both treatments, but surgical approach has been associated with higher cost and risks of causing unnecessary adverse effects. Searching in Epistemonikos database, which is maintained by screening 30 databases, we identified 14 systematic reviews including 25 randomized trials. We combined the evidence using meta-analysis and generated a summary of findings table following the GRADE approach. We concluded that operative treatment may decrease the risk of neurologic impairment, but in turn, could increase the risk of general complications. It is unclear whether there are differences in pain reduction, improvement in function and quality of life, need for subsequent surgery or radiographic progression of kyphosis in both groups.


Las fracturas toracolumbares tipo burst representan hasta el 17% de las fracturas de columna.Se ha planteado tanto el tratamiento conservador como el quirúrgico para este tipo de fracturas, observando buenos resultados funcionales con ambos, pero con un mayor costo y riesgo de producir efectos adversos con la cirugía. Utilizando la base de datos Epistemonikos, la cual es mantenida mediante búsquedas en 30 bases de datos, identificamos seis revisiones sistemáticas que en conjunto incluyen cuatro estudios aleatorizados. Realizamos un metanálisis y tablas de resumen de los resultados utilizando el método GRADE. Concluimos que el tratamiento quirúrgico podría disminuir el riesgo de deterioro neurológico pero con un mayor riesgo de complicaciones generales. Concluimos que no está claro si existen diferencias en dolor, funcionalidad, calidad de vida, necesidad de reintervención o progresión radiográfica de la cifosis entre el tratamiento conservador y el quirúrgico.


Assuntos
Dor/etiologia , Qualidade de Vida , Fraturas da Coluna Vertebral/terapia , Bases de Dados Factuais , Humanos , Vértebras Lombares/lesões , Complicações Pós-Operatórias/epidemiologia , Ensaios Clínicos Controlados Aleatórios como Assunto , Fraturas da Coluna Vertebral/cirurgia , Vértebras Torácicas/lesões , Resultado do Tratamento
19.
Food Chem ; 196: 733-8, 2016 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-26593548

RESUMO

Forty-five women (35-45 year) were randomly assigned to three iron (Fe) absorption sub-studies, which measured the effects of dietary animal proteins on the absorption of heme Fe. Study 1 was focused on heme, red blood cell concentrate (RBCC), hemoglobin (Hb), RBCC+beef meat; study 2 on heme, heme+fish, chicken, and beef; and study 3 on heme and heme+purified animal protein (casein, collagen, albumin). Study 1: the bioavailability of heme Fe from Hb was similar to heme only (∼13.0%). RBCC (25.0%) and RBCC+beef (21.3%) were found to be increased 2- and 1.6-fold, respectively, when compared with heme alone (p<0.05). Study 2: the bioavailability from heme alone (10.3%) was reduced (p<0.05) when it was blended with fish (7.1%) and chicken (4.9%), however it was unaffected by beef. Study 3: casein, collagen, and albumin did not affect the bioavailability of Fe. Proteins from animal source foods and their digestion products did not enhance heme Fe absorption.


Assuntos
Proteínas Alimentares/metabolismo , Heme/metabolismo , Ferro/metabolismo , Carne/análise , Proteínas/metabolismo , Adulto , Animais , Disponibilidade Biológica , Bovinos , Galinhas , Proteínas Alimentares/análise , Digestão , Feminino , Peixes , Heme/análise , Humanos , Pessoa de Meia-Idade , Proteínas/análise
20.
Medwave ; 15 Suppl 3: e6315, 2015 Nov 16.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-26610278

RESUMO

There are several nonsurgical alternatives to treat radicular pain in degenerative lumbar spinal stenosis. Epidural steroid injections have been used for several decades, but the different studies have shown variable effects. Searching in Epistemonikos database, which is maintained by screening 30 databases, we identified nine systematic reviews including seven pertinent randomized controlled trials. We concluded epidural steroid injection probably leads to little or no effect on reducing radicular pain of spinal stenosis.


Existe una variada cantidad de alternativas no quirúrgicas para tratar el dolor radicular producido por la raquiestenosis lumbar degenerativa. Los corticoides epidurales se utilizan desde hace varias décadas, sin embargo la eficacia reportada en la literatura es muy variable. Utilizando la base de datos Epistemonikos, la cual es mantenida mediante búsquedas en 30 bases de datos, identificamos nueve revisiones sistemáticas que en conjunto incluyen siete estudios aleatorizados. Realizamos un metanálisis y tablas de resumen de los resultados utilizando el método GRADE. Concluimos que la inyección de corticoides epidurales probablemente tiene poco o nulo efecto en reducir el dolor radicular por estenorraquis.


Assuntos
Glucocorticoides/administração & dosagem , Radiculopatia/tratamento farmacológico , Estenose Espinal/tratamento farmacológico , Glucocorticoides/uso terapêutico , Humanos , Injeções Epidurais , Vértebras Lombares , Radiculopatia/etiologia , Ensaios Clínicos Controlados Aleatórios como Assunto , Estenose Espinal/patologia
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