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1.
New Phytol ; 233(2): 890-904, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34657283

RESUMEN

The Pseudomonas syringae DC3000 type III effector HopAM1 suppresses plant immunity and contains a Toll/interleukin-1 receptor (TIR) domain homologous to immunity-related TIR domains of plant nucleotide-binding leucine-rich repeat receptors that hydrolyze nicotinamide adenine dinucleotide (NAD+ ) and activate immunity. In vitro and in vivo assays were conducted to determine if HopAM1 hydrolyzes NAD+ and if the activity is essential for HopAM1's suppression of plant immunity and contribution to virulence. HPLC and LC-MS were utilized to analyze metabolites produced from NAD+ by HopAM1 in vitro and in both yeast and plants. Agrobacterium-mediated transient expression and in planta inoculation assays were performed to determine HopAM1's intrinsic enzymatic activity and virulence contribution. HopAM1 is catalytically active and hydrolyzes NAD+ to produce nicotinamide and a novel cADPR variant (v2-cADPR). Expression of HopAM1 triggers cell death in yeast and plants dependent on the putative catalytic residue glutamic acid 191 (E191) within the TIR domain. Furthermore, HopAM1's E191 residue is required to suppress both pattern-triggered immunity and effector-triggered immunity and promote P. syringae virulence. HopAM1 manipulates endogenous NAD+ to produce v2-cADPR and promote pathogenesis. This work suggests that HopAM1's TIR domain possesses different catalytic specificity than other TIR domain-containing NAD+ hydrolases and that pathogens exploit this activity to sabotage NAD+ metabolism for immune suppression and virulence.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas Bacterianas/metabolismo , NAD/metabolismo , Enfermedades de las Plantas/microbiología , Pseudomonas syringae/fisiología , Receptores de Interleucina-1/metabolismo , Virulencia
2.
Rev Gastroenterol Peru ; 42(4): 261-263, 2022.
Artículo en Español | MEDLINE | ID: mdl-36746468

RESUMEN

We present the case of a 64-year-old man who, after a first episode of acute pancreatitis, was readmitted 20 days later due to severe epigastric pain and later an episode of upper gastrointestinal bleeding in the form of hematemesis and melena with hemodynamic instability. An urgent gastroscopy was performed at that time, revealing a probable gastrointestinal fistula in the duodenal bulb with an adherent clot without active bleeding at that time, so an urgent CT angiography was performed that revealed a necrotic peripancreatic collection with the presence of active bleeding inside from the pancreatoduodenal artery. Urgent arteriography identified an image compatible with arterial pseudoaneurysm dependent on the pancreaticoduodenal artery branch, which was successfully embolized. Unfortunately, the patient died a few hours later as a result of septic shock secondary to an infected pancreatic collection.


Asunto(s)
Aneurisma Falso , Pancreatitis , Masculino , Humanos , Persona de Mediana Edad , Pancreatitis/complicaciones , Aneurisma Falso/complicaciones , Aneurisma Falso/diagnóstico , Enfermedad Aguda , Hemorragia Gastrointestinal/complicaciones , Duodeno/irrigación sanguínea
3.
Sex Transm Dis ; 47(3): 162-164, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-31876865

RESUMEN

BACKGROUND: The early identification of the Chlamydia trachomatis variants that cause lymphogranuloma venereum (LGV) is very important to establish an adequate antibiotic treatment. This identification should be made with molecular techniques that are easy to perform and accessible to most microbiology laboratories. The objective of this study was to evaluate 2 real-time polymerase chain reaction (PCR)-based assay (VIASURE Haemophilus ducreyi + C. trachomatis (LGV) real-time PCR detection kit and the Allplex Genital ulcer Assay) for the detection of LGV in rectal samples. MATERIALS AND METHODS: Prospective study on positive rectal samples for C. trachomatis. All samples were processed in parallel by both tests. As a molecular reference method and to solve possible discrepancies between both assays, a PCR-based restriction fragment length polymorphism analysis of the major outer membrane protein gene (omp1) was used. RESULTS: In total, we detected 157 positive rectal samples for C. trachomatis, of which 36 were identified as LGV by PCR-based restriction fragment length polymorphism analysis. The positive percent agreement, negative percent agreement, and overall percent agreement were 88.9%, 100%, and 97.3%, respectively, for the Allplex Genital ulcer assay and 91.6%, 100%, and 97.1%, respectively, for the VIASURE assay. In the direct comparison between the Seegene assay and the VIASURE assay, we obtained a kappa concordance index of 0.98 between both tests. CONCLUSIONS: According to the results obtained, both tests could be used for the detection of LGV in rectal samples.


Asunto(s)
Chlamydia trachomatis , Linfogranuloma Venéreo , Técnicas de Diagnóstico Molecular , Reacción en Cadena en Tiempo Real de la Polimerasa , Chlamydia trachomatis/genética , Humanos , Linfogranuloma Venéreo/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/normas , Estudios Prospectivos , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Recto/microbiología
4.
Enferm Infecc Microbiol Clin ; 35(6): 364-366, 2017.
Artículo en Inglés, Español | MEDLINE | ID: mdl-26415750

RESUMEN

INTRODUCTION: A variable percentage of samples analysed using the Cobas 4800 assay can give an invalid result by PCR inhibition or erroneous due to incorrect DNA extraction with the Cobas 4800 CT/NG test. METHOD: An analysis was performed using the vortex agitation and dilution protocol on the original sample (swab or urine) for a total of 116 samples. In order to analyse the sensitivity of this method, 100 samples (swabs and urine) with known results were retested. RESULTS: A total of 98.3% (114/116) of the samples analysed were resolved with this protocol with 100% agreement after reviewing clinical data, Gram stain, and other samples analysed in parallel from the same patient. DISCUSSION: The data indicate no loss of sensitivity with this protocol; thus Cobas 4800 users could use this method without the need for alternative methods.


Asunto(s)
Técnicas de Tipificación Bacteriana/instrumentación , Chlamydia trachomatis/aislamiento & purificación , ADN Bacteriano/análisis , Neisseria gonorrhoeae/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/instrumentación , Manejo de Especímenes/métodos , Técnicas de Tipificación Bacteriana/métodos , Portador Sano/microbiología , Cuello del Útero/microbiología , Infecciones por Chlamydia/microbiología , Chlamydia trachomatis/genética , ADN Bacteriano/aislamiento & purificación , Exudados y Transudados/microbiología , Femenino , Gonorrea/microbiología , Humanos , Masculino , Neisseria gonorrhoeae/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Recto/microbiología , Sensibilidad y Especificidad , Orina/microbiología
6.
Enferm Infecc Microbiol Clin ; 33(10): 642-5, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-25858681

RESUMEN

INTRODUCTION: A prospective study was designed to assess the performance of the new CT OligoGen kit and the cobas 4800 assay for detection of Chlamydia trachomatis. METHODS: A set of samples that included urine samples (n=212), endocervical (n=167), rectal (n=53), pharyngeal (n=7) and urethral swabs (n=3). The samples were sent from a regional sexually transmitted diseases (STD) clinic in Seville, Spain, and were collected from 261 men and 181 women. Discordant results were re-analyzed and clinical data and other tests were reviewed in order to resolve them. RESULTS: Sensitivity, specificity, positive predicative value (PPV), negative predictive value (NPV) and kappa value for C. trachomatis detection using the CT OligoGen kit were 98.5%, 100%, 100%, 95.4% and 0.97, respectively. CONCLUSIONS: This new kit had a high sensitivity, specificity, PPV and NPV for C. trachomatis, therefore the performance profile confirms the usefulness and reliable results of this new assay.


Asunto(s)
Infecciones por Chlamydia/microbiología , Chlamydia trachomatis/aislamiento & purificación , Cromatografía de Afinidad/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Adulto , Enfermedades Asintomáticas , Cuello del Útero/microbiología , Infecciones por Chlamydia/diagnóstico , ADN Bacteriano/análisis , Femenino , Humanos , Masculino , Faringe/microbiología , Valor Predictivo de las Pruebas , Estudios Prospectivos , Juego de Reactivos para Diagnóstico , Recto/microbiología , Sensibilidad y Especificidad , Uretra/microbiología , Orina/microbiología , Adulto Joven
7.
Angiogenesis ; 17(4): 851-66, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-24728929

RESUMEN

Disorders in skin wound healing are a major health problem that requires the development of innovative treatments. The use of biomaterials as an alternative of skin replacement has become relevant, but its use is still limited due to poor vascularization inside the scaffolds, resulting in insufficient oxygen and growth factors at the wound site. In this study, we have developed a cell-based wound therapy consisting of the application of collagen-based dermal scaffolds containing mesenchymal stem cells from Wharton's jelly (WJ-MSC) in an immunocompetent mouse model of angiogenesis. From our comparative study on the secretion profile between WJ-MSC and adipose tissue-derived MSC, we found a stronger expression of several well-characterized growth factors, such as VEGF-A, angiopoietin-1 and aFGF, which are directly linked to angiogenesis, in the culture supernatant of WJ-MSC, both on monolayer and 3D culture conditions. WJ-MSC proved to be angiogenic both in vitro and in vivo, through tubule formation and CAM assays, respectively. Moreover, WJ-MSC consistently improved the healing response in vivo in a mouse model of human-like dermal repair, by triggering angiogenesis and further providing a suitable matrix for wound repair, without altering the inflammatory response in the animals. Since these cells can be easily isolated, cultured with high expansion rates and cryopreserved, they represent an attractive stem cell source for their use in allogeneic cell transplant and tissue engineering.


Asunto(s)
Células Madre Mesenquimatosas/citología , Neovascularización Patológica , Regeneración/fisiología , Piel/metabolismo , Gelatina de Wharton/química , Adipocitos/citología , Animales , Materiales Biocompatibles , Proliferación Celular , Pollos , Membrana Corioalantoides , Criopreservación , Medios de Cultivo Condicionados , Citometría de Flujo , Humanos , Inflamación , Masculino , Ratones , Ratones Endogámicos BALB C , Osteogénesis , Proteoma , Piel/patología , Ingeniería de Tejidos , Cordón Umbilical/patología , Cicatrización de Heridas
8.
Enferm Infecc Microbiol Clin (Engl Ed) ; 41(2): 103-106, 2023 02.
Artículo en Inglés | MEDLINE | ID: mdl-35906175

RESUMEN

BACKGROUND: Mycoplasma genitalium (M. genitalium) is an emerging sexually transmitted pathogen of increasing importance. The objective of this study was to compare two tests for the detection of M. genitalium; the Aptima® MG test (Hologic® Inc., San Diego, CA) and the Cobas® TV/MG test (Roche® Diagnostics, Mannheim, Germany). METHODS: This is a prospective descriptive study where a total of 489 genital and extragenital samples were analyzed in parallel and in random order by both systems. The samples were collected from patients attending the Sexually Transmitted Infections Center in Seville and the Infectious Diseases consultation of the Virgen de Valme Hospital. RESULTS: The overall agreement between both trials was very good (k > 0.91). The sensitivity and specificity of the Aptima® MG test were 100% and 98.7% respectively for the Cobas® TV/MG test. CONCLUSION: Both systems showed excellent performance for the detection of M. genitalium.


Asunto(s)
Infecciones por Mycoplasma , Mycoplasma genitalium , Enfermedades de Transmisión Sexual , Humanos , Mycoplasma genitalium/genética , Enfermedades de Transmisión Sexual/diagnóstico , Conducta Sexual , Infecciones por Mycoplasma/diagnóstico , Sensibilidad y Especificidad
9.
Saudi J Biol Sci ; 30(1): 103496, 2023 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-36419924

RESUMEN

The study of macroalgae antimicrobial agents is limited to Mexico and scarce in the Veracruzano Reef System (SAV). It is necessary to devote efforts towards this field of applied phycology. The aim was to evaluate the antimicrobial activity of some phyla of Rhodophyta, Chlorophyta and Ochrophyta from SAV. Methanolic extracts from 23 marine macroalgae species (7 Chlorophyta, 4 Phaeophyta and 12 Rhodophyta) from the Veracruzano Reef System (SAV) (Mexico) were evaluated for antimicrobial activity. Antibacterial and antifungal activity were assessed by agar diffusion and agar dilution methods. The differences between mean values obtained for experimental groups was done by analysis of variance (ANOVA multifactorial model), p-values of 0.001 or less were considered statistically significant. Two new records are recognized for SAV (Laurencia gracilis and Sebdenia flabellata) and Compsothamnion thuioides for the Gulf of Mexico coasts. 16 species showed antibacterial activity, of which Caulerpa sertularioides, Ulva lactuca and Laurencia obtuse had significant activity on Gram-positive bacteria. 43.7% belong to the phyla Chlorophyta (7 species), 50% Rhodophyta (8 species) and 6.25% Ochrophyta (1 species). This indicates that the extracts of the algae of the Rhodophyta and Chlorophyta are the ones that showed the greatest activity. Regarding the yeasts, 16.6% of the total algae collected were active in the different yeast strains. 43.7% belongs to Chlorophyta species and for Rhodophyta were 60%. The macroalgae with the highest antifungal activity were: Cymopolia barbata, Ulva lactuca and Laurencia gracilis. The macroalgae of the Veracruzano Reef System present antimicrobial activity. This study is the first investigation of macroalgae's bioactive components from SAV, where they could be sources for future medical applications.

10.
Enferm Infecc Microbiol Clin ; 30(9): 542-8, 2012 Nov.
Artículo en Español | MEDLINE | ID: mdl-22365617

RESUMEN

OBJECTIVE: The objective of this study was the analysis of the prevalence and type of primary resistance to antiretroviral drugs in patients diagnosed with HIV infection, and to determine the most appropriate empirical treatment to obtain a virological and immunological response. PATIENTS AND METHODS: An observational analysis of patients with a de novo diagnosis of HIV infection during the period 2008-2010. Clinical, immunological and virological characteristics, including genotype analysis of resistance to antiretrovirals, were considered as independent variables. The dependent variable was an undetectable HIV viral load after six months of treatment. Data are provided as median (interquartile range) and absolute number (percentage). RESULTS: Seventy-three patients with a de novo diagnosis of HIV infection were included [53 males (73%); 36 (30-46) years-old; prior use of intravenous drugs: 5 patients (7%); hepatitis C virus co-infection: 13 individuals (18%)]. Ten patients (14%) showed symptoms attributable to acute HIV infection. A CD4+ T cell count lower than 350 mm(3) was detected in a 37% (n=27) of all patients. The initiation of antiretroviral therapy followed the GESIDA recommendations (no therapy: 20 patients; tenofovir+emtricitabine+efavirenz: 28 patients; abacavir+lamivudine+efavirenz: 1 patient; tenofovir+emtricitabine+protease inhibitors: 5 patients; abacavir+lamivudine+protease inhibitors: 1 patient; 18 patients were lost in the follow-up). After starting antiretroviral therapy, the resistance analyses detected the existence of primary resistance to antiretrovirals in 12.7% (confidence interval 95%: 3-22) of the patients, distributed as follows: isolated resistance to, nucleosides was detected in 2% (M184V), to nevirapine/efavirenz in 9% (K103N), and combined resistance to nucleosides and non-nucleosides in 2%; there were no cases of resistance to protease inhibitors. Consequently, antiretroviral therapy was changed in 5 (14%) out of 35 patients, attaining an undetectable HIV viral load at 6 months in all of them. The primary resistance to antiretrovirals was not related with epidemiological, virological (including infection by non B subtype) or immunological variables. CONCLUSIONS: In the present study, a change in the epidemiological pattern of de novo diagnosis of HIV infection in our area has been observed. The existence of resistance mutations in more than 5% of the new cases is noteworthy. This finding must be considered in order to establish the rules of empirical treatment in our area.


Asunto(s)
Antirretrovirales/farmacología , Antirretrovirales/uso terapéutico , Infecciones por VIH/tratamiento farmacológico , VIH/efectos de los fármacos , Adulto , Combinación de Medicamentos , Farmacorresistencia Viral , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven
11.
Diagn Microbiol Infect Dis ; 104(4): 115798, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36215941

RESUMEN

The detection of resistance without the need for culture is essential to establish a guided treatment against Neisseria gonorrhoeae (NG) infections. We evaluated the VIASURE Neisseria gonorrhoeae ciprofloxacin resistant Real Time PCR Detection Kit (CerTest Biotec S.L, Zaragoza, Spain) for the simultaneous identification and direct detection of ciprofloxacin susceptibility in 88 NG isolates and 133 positive NG clinical samples of different anatomical location. The sensitivity for NG detection was 93.2% and the specificity 100%. The sensitivity of the test to characterize resistance/susceptibility to ciprofloxacin was (96.5%). In conclusion, the test evaluated is suitable for use to establish a targeted therapy with oral ciprofloxacin in case of not detecting resistance.


Asunto(s)
Gonorrea , Neisseria gonorrhoeae , Humanos , Neisseria gonorrhoeae/genética , Ciprofloxacina/farmacología , Pruebas de Sensibilidad Microbiana , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Gonorrea/diagnóstico , Gonorrea/tratamiento farmacológico , Farmacorresistencia Bacteriana
12.
Pharmaceutics ; 14(11)2022 Nov 19.
Artículo en Inglés | MEDLINE | ID: mdl-36432710

RESUMEN

One of the most important threats to public health is the appearance of multidrug-resistant pathogenic bacteria, since they are the cause of a high number of deaths worldwide. Consequently, the preparation of new effective antibacterial agents that do not generate antimicrobial resistance is urgently required. We report on the synthesis of new linear cationic antibacterial polytriazoles that could be a potential source of new antibacterial compounds. These polymers were prepared by thermal- or copper-catalyzed click reactions of azide and alkyne functions. The antibacterial activity of these materials can be modulated by varying the size or nature of their side chains, as this alters the hydrophilic/hydrophobic balance. Antibacterial activity was tested against pathogens of the ESKAPE group. The P3TD polymer, which has butylated side chains, was found to have the highest bactericidal activity. The toxicity of selected polytriazoles was investigated using human red blood cells and a human gingival fibroblast cell line. The propensity of prepared polytriazoles to induce resistance in certain bacteria was studied. Some of them were found to not produce resistance in methicillin-resistant Staphylococcus aureus or Pseudomonas aeruginosa. The interaction of these polytriazoles with the Escherichia coli membrane produces both depolarization and disruption of the membrane.

13.
Microbes Infect ; 24(8): 105015, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35623592

RESUMEN

AIM: To report two atypical inclusion conjunctivitis cases due to Chlamydia trachomatis in young adults. METHOD: Transcription mediated amplification for C. trachomatis was performed using Aptima Combo 2 Assay (Hologic, Spain). RESULTS: The first patient was managed as an orbital disorder because he had unilateral location, and ptosis was observed. Orbital nuclear magnetic resonance revealed normal results, and conjunctival biopsy did not indicate significant results. For the second patient, thyroid eye disease was suspected, but the orbital nuclear magnetic resonance revealed normal results. Conjunctival exudate samples were collected and sent to the Microbiology Laboratory where C. trachomatis was confirmed. Both patients demonstrated a great improvement with oral azithromycin 1 g. CONCLUSION: Inclusion conjunctivitis could present as unspecified unilateral or bilateral chronic conjunctivitis. Thus, suspecting it would be important in order to prevent spread and wasting diagnostic resources.


Asunto(s)
Infecciones por Chlamydia , Conjuntivitis de Inclusión , Gonorrea , Masculino , Adulto Joven , Humanos , Chlamydia trachomatis/genética , Conjuntivitis de Inclusión/diagnóstico , Conjuntivitis de Inclusión/microbiología , Infecciones por Chlamydia/diagnóstico , Infecciones por Chlamydia/tratamiento farmacológico , Infecciones por Chlamydia/microbiología , Gonorrea/diagnóstico , Gonorrea/microbiología , Hospitales
14.
Obes Res Clin Pract ; 16(6): 533-535, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36283924

RESUMEN

BACKGROUND: The 30″ sit to stand test is a submaximal exercise test that assesses functional capacity and it has been validated for various pathologies. Although it has been used in individuals with obesity, its reproducibility in this population has not yet been determined. The main objective of this study was to determine the reproducibility and safety of the 30″ sit to stand test in individuals with overweight or obesity and with cardiovascular risk factors. METHODS: A cross-sectional study was performed. Individuals with obesity or overweight who also presented cardiovascular risk factors were evaluated with the 30″ sit to stand test. The reproducibility and safety of the 30" sit to stand test were determined, as well as its association with other functional tests and anthropometric characteristics. RESULTS: 59 individuals (27 men, 32 women) with obesity or overweight and cardiovascular risk factors, aged 57.93 (9.62) years, were included in the study. The 30″ sit to stand test showed good overall reproducibility (0.907 ICC) and significant correlation with the 6-minute walk test, handgrip strength test, body fat percentage and waist - height index, with a similar hemodynamic response to the 6-minute walk test. CONCLUSION: The 30" sit to stand test is a highly reproducible and safe test for individuals with obesity and cardiovascular risk factors, with a significant correlation to anthropometric characteristics and other functional tests regularly used for the evaluation of individuals with obesity.


Asunto(s)
Enfermedades Cardiovasculares , Sobrepeso , Masculino , Humanos , Femenino , Sobrepeso/complicaciones , Fuerza de la Mano , Estudios Transversales , Reproducibilidad de los Resultados , Enfermedades Cardiovasculares/etiología , Factores de Riesgo , Obesidad/complicaciones , Factores de Riesgo de Enfermedad Cardiaca
15.
Antibiotics (Basel) ; 10(3)2021 Mar 12.
Artículo en Inglés | MEDLINE | ID: mdl-33809081

RESUMEN

In Tehuacán-Cuicatlán valley (Mexico), studies have been carried out on the essential oils of medicinal plants with antimicrobial activity and it was found that they present compounds in common such as: α-pinene, ß-pinene, carvacrol, eugenol, limonene, myrcene, ocimene, cineole, methyl salicylate, farnesene, and thymol. The goal of this study was to assess the antimicrobial activity of essential oils' compounds. The qualitative evaluation was carried out by the Kirby Baüer agar diffusion technique in Gram-positive bacteria (11 strains), Gram-negative bacteria (18 strains), and yeasts (8 strains). For the determination of the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), the agar dilution method was used. All the evaluated compounds presented antimicrobial activity. The compounds eugenol and carvacrol showed the largest inhibition zones. Regarding yeasts, the compounds ocimene, cineole, and farnesene did not show any activity. The compounds eugenol, carvacrol, and thymol presented the lowest MIC; bactericidal effect was observed at MIC level for S. aureus 75MR, E. coli 128 MR, and C albicans CUSI, for different compounds, eugenol, carvacrol, and thymol. Finally, this study shows that the essential oils of plants used by the population of Tehuacán-Cuicatlán valley share compounds and some of them have antibacterial and fungicidal activity.

16.
Front Cell Dev Biol ; 8: 592442, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33511115

RESUMEN

Cancer cell adhesion to the vascular endothelium is an important step in tumor metastasis. Thy-1 (CD90), a cell adhesion molecule expressed in activated endothelial cells, has been implicated in melanoma metastasis by binding to integrins present in cancer cells. However, the signaling pathway(s) triggered by this Thy-1-Integrin interaction in cancer cells remains to be defined. Our previously reported data indicate that Ca2+-dependent hemichannel opening, as well as the P2X7 receptor, are key players in Thy-1-αVß3 Integrin-induced migration of reactive astrocytes. Thus, we investigated whether this signaling pathway is activated in MDA-MB-231 breast cancer cells and in B16F10 melanoma cells when stimulated with Thy-1. In both cancer cell types, Thy-1 induced a rapid increase in intracellular Ca2+, ATP release, as well as cell migration and invasion. Connexin and Pannexin inhibitors decreased cell migration, implicating a requirement for hemichannel opening in Thy-1-induced cell migration. In addition, cell migration and invasion were precluded when the P2X7 receptor was pharmacologically blocked. Moreover, the ability of breast cancer and melanoma cells to transmigrate through an activated endothelial monolayer was significantly decreased when the ß3 Integrin was silenced in these cancer cells. Importantly, melanoma cells with silenced ß3 Integrin were unable to metastasize to the lung in a preclinical mouse model. Thus, our results suggest that the Ca2+/hemichannel/ATP/P2X7 receptor-signaling axis triggered by the Thy-1-αVß3 Integrin interaction is important for cancer cell migration, invasion and transvasation. These findings open up the possibility of therapeutically targeting the Thy-1-Integrin signaling pathway to prevent metastasis.

17.
Biochem Pharmacol ; 177: 113941, 2020 07.
Artículo en Inglés | MEDLINE | ID: mdl-32240650

RESUMEN

In advanced stages of cancer disease, caveolin-1 (CAV1) expression increases and correlates with increased migratory and invasive capacity of the respective tumor cells. Previous findings from our laboratory revealed that specific ECM-integrin interactions and tyrosine-14 phosphorylation of CAV1 are required for CAV1-enhanced melanoma cell migration, invasion and metastasis in vivo. In this context, CAV1 phosphorylation on tyrosine-14 mediated by non-receptor Src-family tyrosine kinases seems to be important; however, the effect of Src-family kinase inhibitors on CAV1-enhanced metastasis in vivo has not been studied. Here, we evaluated the effect of CAV1 and c-Abl overexpression, as well as the use of the Src-family kinase inhibitors, PP2 and dasatinib (more specific for Src/Abl) in lung metastasis of B16F10 melanoma cells. Overexpression of CAV1 and c-Abl enhanced CAV1 phosphorylation and the metastatic potential of the B16F10 murine melanoma cells. Alternatively, treatment with PP2 or dasatinib for 2 h reduced CAV1 tyrosine-14 phosphorylation and levels recovered fully within 12 h of removing the inhibitors. Nonetheless, pre-treatment of cells with these inhibitors for 2 h sufficed to prevent migration, invasion and trans-endothelial migration in vitro. Importantly, the transient decrease in CAV1 phosphorylation by these kinase inhibitors prevented early steps of CAV1-enhanced lung metastasis by B16F10 melanoma cells injected into the tail vein of mice. In conclusion, this study underscores the relevance of CAV1 tyrosine-14 phosphorylation by Src-family kinases during the first steps of the metastatic sequence promoted by CAV1. These findings open up potential options for treatment of metastatic tumors in patients in which Src-family kinase activation and CAV1 overexpression favor dissemination of cancer cells to secondary sites.


Asunto(s)
Caveolina 1/metabolismo , Dasatinib/farmacología , Neoplasias Pulmonares/secundario , Melanoma Experimental/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , Pirimidinas/farmacología , Neoplasias Cutáneas/metabolismo , Familia-src Quinasas/antagonistas & inhibidores , Animales , Caveolina 1/genética , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Dasatinib/uso terapéutico , Femenino , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/patología , Masculino , Melanoma Experimental/patología , Ratones , Ratones Endogámicos C57BL , Neuropéptidos/metabolismo , Fosforilación/efectos de los fármacos , Inhibidores de Proteínas Quinasas/uso terapéutico , Pirimidinas/uso terapéutico , Neoplasias Cutáneas/patología , Transfección , Tirosina/metabolismo , Proteína de Unión al GTP rac1/metabolismo
18.
Genes (Basel) ; 11(2)2020 01 25.
Artículo en Inglés | MEDLINE | ID: mdl-31991774

RESUMEN

Presented here are five members of a family that was ascertained from an isolated, consanguineous, indigenous Amerindian community in Colombia that was affected with calpain 3-related, limb-girdle muscular dystrophy type R1. These patients are homozygous for a unique and novel deletion of four bases (TGCC) in exon 3 of the calpain 3 gene (CAPN3) (NM_000070.2; NP_000061.1) (g.409_412del). The mutation site occurs at the CysPc protein domain, triggering a modified truncated protein structure and affecting motifs within the calpain-like thiol protease family (peptidase family C2) region. The patients reported here developed a very severe phenotype with primary contractures, spinal rigidity in the early stages of the disease, and bilateral talipes equinovarus (clubfoot) in the most affected patients who had the selective involvement of their extremities' distal muscles in a way that resembled Emery-Dreifuss syndrome. We recommend mandatory screening for calpainopathy in all patients with an Emery-Dreifuss-like syndrome or those presenting a non-congenital illness with primary contractures and who, because of other data, are suspected of having muscular dystrophy.


Asunto(s)
Calpaína/genética , Proteínas Musculares/genética , Distrofia Muscular de Cinturas/genética , Distrofias Musculares/genética , Adolescente , Adulto , Niño , Exones/genética , Familia , Femenino , Homocigoto , Humanos , Masculino , Distrofia Muscular de Cinturas/metabolismo , Mutación/genética , Linaje , Fenotipo , Eliminación de Secuencia/genética
19.
Front Cell Dev Biol ; 7: 132, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31428610

RESUMEN

Thy-1/CD90 is a glycoprotein attached to the outer face of the plasma membrane with various functions, which depend on the context of specific physiological or pathological conditions. Many of these reported functions for Thy-1/CD90 arose from studies by our group, which identified the first ligand/receptor for Thy-1/CD90 as an integrin. This finding initiated studies directed toward unveiling the molecular mechanisms that operate downstream of Thy-1/CD90 activation, and its possible interaction with proteins in the membrane plane to regulate their function. The association of Thy-1/CD90 with a number of cell surface molecules allows the formation of extra/intracellular multiprotein complexes composed of various ligands and receptors, extracellular matrix proteins, intracellular signaling proteins, and the cytoskeleton. The complexes sense changes that occur inside and outside the cells, with Thy-1/CD90 at the core of this extracellular molecular platform. Molecular platforms are scaffold-containing microdomains where key proteins associate to prominently influence cellular processes and behavior. Each component, by itself, is less effective, but when together with various scaffold proteins to form a platform, the components become more specific and efficient to convey the messages. This review article discusses the experimental evidence that supports the role of Thy-1/CD90 as a membrane-associated platform (ThyMAP).

20.
Nanomaterials (Basel) ; 9(7)2019 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-31261871

RESUMEN

We report on the design, development, characterization, and a preliminary cellular evaluation of a novel solid material. This material is composed of low-molecular-weight hyaluronic acid (LMWHA) and polyarginine (PArg), which generate aqueous ionic nanocomplexes (INC) that are then freeze-dried to create the final product. Different ratios of LMWHA/PArg were selected to elaborate INC, the size and zeta potential of which ranged from 100 to 200 nm and +25 to -43 mV, respectively. Turbidimetry and nanoparticle concentration analyses demonstrated the high capacity of the INC to interact with increasing concentrations of LMWHA, improving the yield of production of the nanostructures. Interestingly, once the selected formulations of INC were freeze-dried, only those comprising a larger excess of LMWHA could form reproducible sponge formulations, as seen with the naked eye. This optical behavior was consistent with the scanning transmission electron microscopy (STEM) images, which showed a tendency of the particles to agglomerate when an excess of LMWHA was present. Mechanical characterization evidenced low stiffness in the materials, attributed to the low density and high porosity. A preliminary cellular evaluation in a fibroblast cell line (RMF-EG) evidenced the concentration range where swollen formulations did not affect cell proliferation (93-464 µM) at 24, 48, or 72 h. Considering that the reproducible sponge formulations were elaborated following inexpensive and non-contaminant methods and comprised bioactive components, we postulate them with potential for biomedical purposes. Additionally, this systematic study provides important information to design reproducible porous solid materials using ionic nanocomplexes.

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