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Surface modification of drug-loaded particles with polyethylene glycol (PEG) chains is a powerful tool that promotes better transport of therapeutic agents, provides stability, and avoids their detection by the immune system. In this study, we used a new approach to synthesize a biodegradable poly(ester amide) (PEA) and PEGylating surfactant. These were employed to fabricate micro/nanoparticles with a core-shell structure. Nanoparticle (NP)-protein interactions and self-assembling were subsequently studied by synchrotron radiation-based FTIR microspectroscopy (SR-FTIRM) and transmission electron microscopy (TEM) techniques. The core-shell structure was identified using IR absorption bands of characteristic chemical groups. Specifically, the stretching absorption band of the secondary amino group (3300 cm-1) allowed us to identify the poly(ester amide) core, while the band at 1105 cm-1 (C-O-C vibration) was useful to demonstrate the shell structure based on PEG chains. By integration of absorption bands, a 2D intensity map of the particle was built to show a core-shell structure, which was further supported by TEM images.
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Nanopartículas , Poliésteres , Polietilenglicoles , Sincrotrones , Polietilenglicoles/química , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Nanopartículas/química , Poliésteres/química , Microscopía Electrónica de Transmisión/métodos , Amidas/químicaRESUMEN
Bacterial contamination is a hazard in many industries, including food, pharmaceuticals, and healthcare. The availability of a rapid and simple method for detecting this type of contamination in sterile areas enables immediate intervention to avoid or reduce detrimental effects. Among these methods, colorimetric indicators are becoming increasingly popular due to their affordability, ease of use, and quick visual interpretation of the signal. In this article, a bacterial contamination indicator system was designed by incorporating MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) into an electrospun PADAS matrix, which is a biodegradable poly(ester amide) synthesized from L-alanine, 1,12-dodecanediol, and sebacic acid. Uniaxial stress testing, thermogravimetric analysis and scanning electron microscopy were used to examine the mechanical properties, thermal stability, and morphology of the mats, respectively. The capacity for bacterial detection was not only analyzed with agar and broth assays but also by replicating important environmental conditions. Among the MTT concentrations tested in this study (0.2%, 2%, and 5%), it was found that only with a 2% MTT content the designed system produced a color response visible to the naked eye with optimal intensity, a sensitivity limit of 104 CFU/mL, and 86% cell viability, which showed the great potential for its use to detect bacterial contamination. In summary, by means of the process described in this work, it was possible to obtain a simple, low-cost and fast-response bacterial contamination indicator that can be used in mask filters, air filters, or protective clothing.
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Colorimetría , Poliésteres , Sales de Tetrazolio , Sales de Tetrazolio/química , Poliésteres/química , Colorimetría/métodos , Tiazoles/química , Bacterias , HumanosRESUMEN
This review presents a comprehensive update of the biopolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), emphasizing its production, properties, and applications. The overall biosynthesis pathway of PHBV is explored in detail, highlighting recent advances in production techniques. The inherent physicochemical properties of PHBV, along with its degradation behavior, are discussed in detail. This review also explores various blends and composites of PHBV, demonstrating their potential for a range of applications. Finally, the versatility of PHBV-based materials in multiple sectors is examined, emphasizing their increasing importance in the field of biodegradable polymers.
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Poliésteres , Polímeros , Ácido 3-Hidroxibutírico , Poliésteres/química , Ácidos PentanoicosRESUMEN
BACKGROUND: At the end of 2019, a novel coronavirus denominated SARS-CoV-2 rapidly spread through the world causing the pandemic coronavirus disease known as COVID-19. The difference in the inflammatory response against SARS-CoV-2 infection among people living at different altitudes is a variable not yet studied. METHODS: A descriptive cross-sectional study was performed in two Peruvian cities at different altitudes for comparison: Lima and Huaraz. Five important proinflammatory cytokines were measured including: IL-6, IL-2, IL-10, IFN-γ and TNF-α using ELISA assays. RESULTS: A total of 35 COVID-19 patients and 10 healthy subjects were recruited from each study site. The mean levels of IL-6 (p < 0.03) and TNF-α (p < 0.01) were significantly different among the study groups. In the case of IL-6, patients from Lima had a mean level of 16.2 pg/ml (healthy) and 48.3 pg/ml (COVID-19), meanwhile, patients from Huaraz had levels of 67.3 pg/ml (healthy) and 97.9 pg/ml (COVID-19). Regarding TNF-α, patients from Lima had a mean level of 25.9 pg/ml (healthy) and 61.6 pg/ml (COVID-19), meanwhile, patients from Huaraz had levels of 89.0 pg/ml (healthy) and 120.6 pg/ml (COVID-19). The levels of IL-2, IL-10 and IFN-γ were not significantly different in the study groups. CONCLUSION: Patients with COVID-19 residing at high-altitude tend to have higher levels of inflammatory cytokines compared to patients living at sea level, particularly IL-6 and TNF-α. A better understanding of the inflammatory response in different populations can contribute to the implementation of therapeutic and preventive approaches. Further studies evaluating more patients, a greater variety of cytokines and their clinical impact are required.
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Altitud , COVID-19 , Estudios Transversales , Citocinas , Humanos , SARS-CoV-2RESUMEN
Great advances in cancer treatment have been undertaken in the last years as a consequence of the development of new antitumoral drugs able to target cancer cells with decreasing side effects and a better understanding of the behavior of neoplastic cells during invasion and metastasis. Specifically, drug delivery systems (DDS) based on the use of hydroxyapatite nanoparticles (HAp NPs) are gaining attention and merit a comprehensive review focused on their potential applications. These are derived from the intrinsic properties of HAp (e.g., biocompatibility and biodegradability), together with the easy functionalization and easy control of porosity, crystallinity and morphology of HAp NPs. The capacity to tailor the properties of DLS based on HAp NPs has well-recognized advantages for the control of both drug loading and release. Furthermore, the functionalization of NPs allows a targeted uptake in tumoral cells while their rapid elimination by the reticuloendothelial system (RES) can be avoided. Advances in HAp NPs involve not only their use as drug nanocarriers but also their employment as nanosystems for magnetic hyperthermia therapy, gene delivery systems, adjuvants for cancer immunotherapy and nanoparticles for cell imaging.
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Nanopartículas , Neoplasias , Sistemas de Liberación de Medicamentos/métodos , Durapatita/uso terapéutico , Humanos , Nanopartículas/uso terapéutico , Neoplasias/tratamiento farmacológico , Neoplasias/terapia , PorosidadRESUMEN
The preparation, characterization, and controlled release of hydroxyapatite (HAp) nanoparticles loaded with streptomycin (STR) was studied. These nanoparticles are highly appropriate for the treatment of bacterial infections and are also promising for the treatment of cancer cells. The analyses involved scanning electron microscopy, dynamic light scattering (DLS) and Z-potential measurements, as well as infrared spectroscopy and X-ray diffraction. Both amorphous (ACP) and crystalline (cHAp) hydroxyapatite nanoparticles were considered since they differ in their release behavior (faster and slower for amorphous and crystalline particles, respectively). The encapsulated nanoparticles were finally incorporated into biodegradable and biocompatible polylactide (PLA) scaffolds. The STR load was carried out following different pathways during the synthesis/precipitation of the nanoparticles (i.e., nucleation steps) and also by simple adsorption once the nanoparticles were formed. The loaded nanoparticles were biocompatible according to the study of the cytotoxicity of extracts using different cell lines. FTIR microspectroscopy was also employed to evaluate the cytotoxic effect on cancer cell lines of nanoparticles internalized by endocytosis. The results were promising when amorphous nanoparticles were employed. The nanoparticles loaded with STR increased their size and changed their superficial negative charge to positive. The nanoparticles' crystallinity decreased, with the consequence that their crystal sizes reduced, when STR was incorporated into their structure. STR maintained its antibacterial activity, although it was reduced during the adsorption into the nanoparticles formed. The STR release was faster from the amorphous ACP nanoparticles and slower from the crystalline cHAp nanoparticles. However, in both cases, the STR release was slower when incorporated in calcium and phosphate during the synthesis. The biocompatibility of these nanoparticles was assayed by two approximations. When extracts from the nanoparticles were evaluated in cultures of cell lines, no cytotoxic damage was observed at concentrations of less than 10 mg/mL. This demonstrated their biocompatibility. Another experiment using FTIR microspectroscopy evaluated the cytotoxic effect of nanoparticles internalized by endocytosis in cancer cells. The results demonstrated slight damage to the biomacromolecules when the cells were treated with ACP nanoparticles. Both ACP and cHAp nanoparticles were efficiently encapsulated in PLA electrospun matrices, providing functionality and bioactive properties.
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Sistemas de Liberación de Medicamentos/métodos , Nanopartículas/uso terapéutico , Estreptomicina/administración & dosificación , Animales , Antibacterianos/química , Infecciones Bacterianas/tratamiento farmacológico , Células COS , Línea Celular Tumoral , Chlorocebus aethiops , Durapatita/química , Humanos , Nanopartículas/química , Poliésteres/química , Estreptomicina/farmacología , Andamios del Tejido/química , Células VeroRESUMEN
Porous biodegradable scaffolds provide a physical substrate for cells allowing them to attach, proliferate and guide the formation of new tissues. A variety of techniques have been developed to fabricate tissue engineering (TE) scaffolds, among them the most relevant is the thermally-induced phase separation (TIPS). This technique has been widely used in recent years to fabricate three-dimensional (3D) TE scaffolds. Low production cost, simple experimental procedure and easy processability together with the capability to produce highly porous scaffolds with controllable architecture justify the popularity of TIPS. This paper provides a general overview of the TIPS methodology applied for the preparation of 3D porous TE scaffolds. The recent advances in the fabrication of porous scaffolds through this technique, in terms of technology and material selection, have been reviewed. In addition, how properties can be effectively modified to serve as ideal substrates for specific target cells has been specifically addressed. Additionally, examples are offered with respect to changes of TIPS procedure parameters, the combination of TIPS with other techniques and innovations in polymer or filler selection.
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Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Animales , Materiales Biocompatibles/química , Humanos , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Poliésteres/química , Polímeros/química , Porosidad , Solventes/química , TemperaturaRESUMEN
Biominerals formed by DNA and calcium oxalate (CaOx) or hydroxyapatite (HAp), the most important and stable phase of calcium phosphate) have been examined and compared using a synergistic combination of computer simulation and experimental studies. The interest of this comparison stems from the medical observation that HAp- and CaOx-based microcalcifications are frequently observed in breast cancer tissues, and some of their features are used as part of the diagnosis. Molecular dynamics simulations show that (1) the DNA double helix remains stable when it is adsorbed onto the most stable facet of HAp, whereas it undergoes significant structural distortions when it is adsorbed onto CaOx; (2) DNA acts as a template for the nucleation and growth of HAp but not for the mineralization of CaOx; and (3) the DNA double helix remains stable when it is encapsulated inside HAp nanopores, but it becomes destabilized when the encapsulation occurs into CaOx nanopores. Furthermore, CaOx and HAp minerals containing DNA molecules inside and/or adsorbed on the surface have been prepared in the lab by mixing solutions containing the corresponding ions with fish sperm DNA. Characterization of the formed minerals, which has been focused on the identification of DNA using UV-vis spectroscopy, indicates that the tendency to adsorb and, especially, encapsulate DNA is much smaller for CaOx than for HAp, which is in perfect agreement with results from molecular dynamics simulations. Finally, quantum mechanical calculations have been performed to rationalize these results in terms of molecular interactions, evidencing the high affinity of Ca2+ toward oxalate anions in an aqueous environment.
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Materiales Biocompatibles/síntesis química , Oxalato de Calcio/química , ADN/química , Durapatita/química , Adsorción , Materiales Biocompatibles/química , Modelos Moleculares , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
Molecular sensing, water purification and desalination, drug delivery, and DNA sequencing are some striking applications of biomimetic hybrid membranes. These devices take advantage of biomolecules, which have gained excellence in their specificity and efficiency during billions of years, and of artificial materials that load the purified biological molecules and provide technological properties, such as robustness, scalability, and suitable nanofeatures to confine the biomolecules. Recent methodological advances allow more precise control of polymer membranes that support the biomacromolecules, and are expected to improve the design of the next generation of membranes as well as their applicability. In the first section of this review we explain the biological relevance of membranes, membrane proteins, and the classification used for the latter. After this, we critically analyse the different approaches employed for the production of highly selective hybrid membranes, focusing on novel materials made of self-assembled block copolymers and nanostructured polymers. Finally, a summary of the advantages and disadvantages of the different methodologies is presented and the main characteristics of biomimetic hybrid membranes are highlighted.
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Proteínas de Transporte de Membrana/química , Membranas Artificiales , Péptidos/química , Polímeros/químicaRESUMEN
In spite of p-doped conducting polymers having been widely studied in the last decades and many applications having been developed, studies based on n-doped conducting polymers are extremely scarce. This fact is even more evident when it comes to conducting polymers n-doped with polycations, even though polyanions, such as poly(styrenesulfonate), are often used to obtain p-doped conducting polymers. In this work poly(pyridinium-1,4-diyliminocarbonyl-1,4-phenylene-methylene chloride), abbreviated as P(Py-1,4-P), has been used to prepare n-doped poly(3,4-ethylenedioxythiophene) (PEDOT) electrodes by applying a reduction potential to a de-doped PEDOT film in a P(Py-1,4-P) water solution. The utilization of this cationic polyelectrolyte as an n-dopant agent results in drastic superficial changes, as is observed by comparing the morphology, topography and wettability of p-doped, de-doped and n-doped PEDOT. Cytotoxicity, cell adhesion and cell proliferation assays, which have been conducted using epithelial and fibroblast cell lines, show that the amount of P(Py-1,4-P) in the re-doped PEDOT films is below that required to observe a cytotoxic harmful response and that n-doped PEDOT:P(Py-1,4-P) films are biocompatible. The non-specific bacteriostatic properties of n-doped PEDOT:P(Py-1,4-P) films have been demonstrated against E. coli and S. aureus bacteria (Gram-negative and Gram-positive, respectively) using bacterial growth curves and adhesion assays. Although the bacteriostatic effect is in part due to the conducting polymer, as is proved by results for p-doped and de-doped PEDOT, the incorporation of P(Py-1,4-P) through the re-doping process greatly enhances this antimicrobial behaviour. Thus, only a small concentration of this cationic polyelectrolyte (â¼0.1 mM) is needed to inhibit bacterial growth.
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BACKGROUND: Acute respiratory infections (ARIs) represent an important cause of morbidity and mortality in children, remaining a major public health concern, especially affecting children under 5 years old from low-income countries. Unfortunately, information regarding their epidemiology is still limited in Peru. METHODS: A secondary data analysis was performed from a previous cross-sectional study conducted in children with a probable diagnosis of Pertussis from January 2010 to July 2012. All samples were analyzed via Polymerase Chain Reaction (PCR) for the following etiologies: Influenza-A, Influenza-B, RSV-A, RSV-B, Adenovirus, Parainfluenza 1 virus, Parainfluenza 2 virus, Parainfluenza 3 virus, Mycoplasma pneumoniae and Chlamydia pneumoniae. RESULTS: A total of 288 patients were included. The most common pathogen isolated was Adenovirus (49%), followed by Bordetella pertussis (41%) from our previous investigation, the most prevelant microorganisms were Mycoplasma pneumonia (26%) and Influenza-B (19.8%). Coinfections were reported in 58% of samples and the most common association was found between B. pertussis and Adenovirus (12.2%). CONCLUSIONS: There was a high prevalence of Adenovirus, Mycoplasma pneumoniae and other etiologies in patients with a probable diagnosis of pertussis. Despite the presence of persistent cough lasting at least two weeks and other clinical characteristics highly suspicious of pertussis, secondary etiologies should be considered in children under 5 years-old in order to give a proper treatment.
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Infecciones del Sistema Respiratorio/microbiología , Infecciones del Sistema Respiratorio/virología , Tos Ferina/etiología , Infecciones por Adenoviridae/epidemiología , Infecciones por Adenoviridae/etiología , Bordetella pertussis/genética , Bordetella pertussis/aislamiento & purificación , Preescolar , Infecciones por Chlamydophila/epidemiología , Infecciones por Chlamydophila/etiología , Chlamydophila pneumoniae/genética , Chlamydophila pneumoniae/aislamiento & purificación , Tos/microbiología , Estudios Transversales , Femenino , Hospitalización , Humanos , Lactante , Recién Nacido , Gripe Humana/epidemiología , Gripe Humana/etiología , Masculino , Mycoplasma pneumoniae/aislamiento & purificación , Virus de la Parainfluenza 3 Humana/genética , Virus de la Parainfluenza 3 Humana/aislamiento & purificación , Perú/epidemiología , Neumonía por Mycoplasma/epidemiología , Neumonía por Mycoplasma/microbiología , Infecciones del Sistema Respiratorio/epidemiología , Infecciones por Respirovirus/epidemiología , Infecciones por Respirovirus/etiología , Tos Ferina/diagnóstico , Tos Ferina/epidemiologíaRESUMEN
Chloramphenicol (CAM) has been encapsulated into hydroxyapatite nanoparticles displaying different morphologies and crystallinities. The process was based on typical precipitation of solutions containing phosphate and calcium ions and the addition of CAM once the hydroxyapatite nuclei were formed. This procedure favored a disposition of the drug into the bulk parts of the nanoparticles and led to a fast release in aqueous media. Clear antibacterial activity was derived, being slightly higher for the amorphous samples due to their higher encapsulation efficiency. Polylactide (PLA) microfibers incorporating CAM encapsulated in hydroxyapatite nanoparticles were prepared by the electrospinning technique and under optimized conditions. Drug release experiments demonstrated that only a small percentage of the loaded CAM could be delivered to an aqueous PBS medium. This amount was enough to render an immediate bacteriostatic effect without causing a cytotoxic effect on osteoblast-like, fibroblasts, and epithelial cells. Therefore, the prepared scaffolds were able to retain CAM-loaded nanoparticles, being a reservoir that should allow a prolonged release depending on the polymer degradation rate. The studied system may have promising applications for the treatment of cancer since CAM has been proposed as a new antitumor drug.
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Antibacterianos/farmacología , Cloranfenicol/farmacología , Durapatita/química , Poliésteres/química , Animales , Antibacterianos/química , Cápsulas , Línea Celular , Cloranfenicol/química , Chlorocebus aethiops , Escherichia coli/efectos de los fármacos , Nanopartículas , Tamaño de la Partícula , Staphylococcus aureus/efectos de los fármacos , Células VeroRESUMEN
The non-isothermal crystallization of the biodegradable poly(4-hydroxybutyrate) (P4HB) has been studied by means of differential scanning calorimetry (DSC) and polarizing optical microscopy (POM). In the first case, Avrami, Ozawa, Mo, Cazé, and Friedman methodologies were applied. The isoconversional approach developed by Vyazovkin allowed also the determination of a secondary nucleation parameter of 2.10 × 105 K2 and estimating a temperature close to 10 °C for the maximum crystal growth rate. Similar values (i.e., 2.22 × 105 K2 and 9 °C) were evaluated from non-isothermal Avrami parameters. All experimental data corresponded to a limited region where the polymer crystallized according to a single regime. Negative and ringed spherulites were always obtained from the non-isothermal crystallization of P4HB from the melt. The texture of spherulites was dependent on the crystallization temperature, and specifically, the interring spacing decreased with the decrease of the crystallization temperature (Tc). Synchrotron data indicated that the thickness of the constitutive lamellae varied with the cooling rate, being deduced as a lamellar insertion mechanism that became more relevant when the cooling rate increased. POM non-isothermal measurements were also consistent with a single crystallization regime and provided direct measurements of the crystallization growth rate (G). Analysis of the POM data gave a secondary nucleation constant and a bell-shaped G-Tc dependence that was in relative agreement with DSC analysis. All non-isothermal data were finally compared with information derived from previous isothermal analyses.
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Poliésteres/química , Algoritmos , Biopolímeros/química , Rastreo Diferencial de Calorimetría , Cristalización , Cinética , Modelos Químicos , TemperaturaRESUMEN
Permanently polarized hydroxyapatite (HAp) particles have been prepared by applying a constant DC of 500â V at 1000 °C for 1â h to the sintered mineral. This process causes important chemical changes, as the formation of OH- defects (vacancies), the disappearance of hydrogenophosphate ions at the mineral surface layer, and structural variations reflected by the increment of the crystallinity. As a consequence, the electrochemical properties and electrical conductivity of the polarized mineral increase noticeably compared with as-prepared and sintered samples. Moreover, these increments remain practically unaltered after several months. In addition, permanent polarization favours significantly the ability of HAp to adsorb inorganic bioadsorbates in comparison with as-prepared and sintered samples. The adsorbates cause a significant increment of the electrochemical stability and electrical conductivity with respect to bare polarized HAp, which may have many implications for biomedical applications of permanently polarized HAp.
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BACKGROUND: Acute febrile illness (AFI) represent a significant health challenge in the Peruvian Amazon basin population due to their diverse etiologies and the unavailability of specific on-site diagnostic methods, resulting in underreporting of cases. In Peru, one of the most endemic regions to dengue and leptospirosis is Madre de Dios, a region also endemic to emergent bacterial etiologic agents of AFI, such as bartonellosis and rickettsiosis, whose prevalence is usually underreported. We aimed to molecularly identify the presence of Leptospira spp., Bartonella bacilliformis, and Rickettsia spp. by Polymerase Chain Reaction in serum samples from patients with AFI from Puerto Maldonado-Madre de Dios in Peru. METHODS: Serum samples from patients with acute febrile illness were analyzed by real-time PCR for detecting the presence of Bartonella bacilliformis, Leptospira spp. and Rickettsia spp. RESULTS: Bartonella bacilliformis was the most prevalent bacteria identified in 21.6% (30/139) of the samples, followed by Leptospira spp. in 11.5% (16/139) and Rickettsia spp. in 6.5% (9/139) of the samples. No co-infections were observed between these bacteria. The most frequent symptoms associated with fever among all groups, were headaches, myalgias, and arthralgias. We found no statistically significant differences in the clinical presentation between patients infected with each bacterium. CONCLUSIONS: In a previous study, we shown the presence of dengue, chikungunya, Zika and oropouche virus. We were able to identify these pathogens in 29.5% of all the samples, with chikungunya and OROV as the most frequently found in 9.4 and 8.6% of all the samples, respectively. In this study we show that B. bacilliformis (21.6%), Leptospira spp. (11.5%) and Rickettsia spp. (6.5%) accounted for the main etiologies of AFI in samples from Puerto Maldonado-Madre de Dios, Perú. Our analysis of their clinical presentation, further shows the importance of implementing more sensitive and specific on-site diagnostic tools in the national surveillance programs.This study confirms that the un-specificity of signs and symptoms is not only associated with arboviral infections, but also with the clinical presentation of endemic bacterial infections.
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Infecciones por Bartonella , Bartonella bacilliformis/genética , Leptospira/genética , Leptospirosis , Infecciones por Rickettsia , Rickettsia/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Infecciones por Bartonella/diagnóstico , Infecciones por Bartonella/epidemiología , Infecciones por Bartonella/genética , Infecciones por Bartonella/microbiología , Bartonella bacilliformis/aislamiento & purificación , Niño , Preescolar , Coinfección , Estudios Transversales , Femenino , Fiebre/diagnóstico , Fiebre/epidemiología , Fiebre/microbiología , Humanos , Lactante , Recién Nacido , Leptospira/aislamiento & purificación , Leptospirosis/diagnóstico , Leptospirosis/epidemiología , Leptospirosis/microbiología , Masculino , Persona de Mediana Edad , Tipificación Molecular , Perú/epidemiología , Prevalencia , Reacción en Cadena en Tiempo Real de la Polimerasa , Rickettsia/aislamiento & purificación , Infecciones por Rickettsia/diagnóstico , Infecciones por Rickettsia/epidemiología , Infecciones por Rickettsia/microbiología , Ríos , Sensibilidad y Especificidad , Adulto JovenRESUMEN
Peptides homing tumor vasculature are considered promising molecular imaging agents for cancer detection at an early stage. In addition to their high binding affinity, improved tissue penetrating ability, and low immunogenicity, they can deliver targeted anticancer drugs, thus expanding therapeutic treatments. Among those, CREKA, a linear peptide that specifically binds to clotted-plasma proteins in tumor vessels, has been recently employed to design bioactive systems able to target different cancer types. Within this context, this paper explores the biorecognition event between CR(NMe)EKA, an engineered CREKA-analog bearing a noncoded amino acid (N-methyl-Glu) that is responsible for its enhanced activity, and clotted-plasma proteins (fibrin and fibrinogen) by nanomechanical detection. Specifically, the tumor-homing peptide was covalently attached via epoxysilane chemistry onto silicon microcantilever chips that acted as sensors during dynamic mode experiments. Before that, each step of the functionalization process was followed by contact angle measurements, interferometry, X-ray photoelectron spectroscopy, and atomic force microscopy, thus revealing the applied protocol as a suitable strategy. The fibrin(ogen)-binding induced by CR(NMe)EKA was detected by the resonance frequency shift of the cantilevers, and a detection limit of 100 ng/mL was achieved for both proteins. Even though further development is required, this work reflects the promising application of emerging technologies capable of assisting in the comprehension of biological interactions and their implications in the biotechnological field. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.
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Técnicas Biosensibles/instrumentación , Fibrina/química , Fibrinógeno/química , Oligopéptidos/química , Análisis por Matrices de Proteínas/instrumentación , Fenómenos Biomecánicos , Técnicas Biosensibles/métodos , Glutamatos/química , Humanos , Límite de Detección , Oligopéptidos/síntesis química , Análisis por Matrices de Proteínas/métodos , Unión Proteica , Ingeniería de Proteínas , Silanos/química , Silicio/química , Propiedades de SuperficieRESUMEN
In spite of the clinical importance of hydroxyapatite (HAp), the mechanism that controls its dissolution in acidic environments remains unclear. Knowledge of such a process is highly desirable to provide better understanding of different pathologies, as for example osteoporosis, and of the HAp potential as vehicle for gene delivery to replace damaged DNA. In this work, the mechanism of dissolution in acid conditions of HAp nanoparticles encapsulating double-stranded DNA has been investigated at the atomistic level using computer simulations. For this purpose, four consecutive (multi-step) molecular dynamics simulations, involving different temperatures and proton transfer processes, have been carried out. Results are consistent with a polynuclear decalcification mechanism in which proton transfer processes, from the surface to the internal regions of the particle, play a crucial role. In addition, the DNA remains protected by the mineral mold and transferred proton from both temperature and chemicals. These results, which indicate that biomineralization imparts very effective protection to DNA, also have important implications in other biomedical fields, as for example in the design of artificial bones or in the fight against osteoporosis by promoting the fixation of Ca(2+) ions.
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ADN/química , Durapatita/química , Iones/química , Nanopartículas/química , ADN/metabolismo , Simulación de Dinámica Molecular , TemperaturaRESUMEN
Homopeptides with 2, 3 and 4 phenylalanine (Phe) residues and capped with fluorenylmethoxycarbonyl and fluorenylmethyl esters at the N-terminus and C-terminus, respectively, have been synthesized to examine their self-assembly capabilities. Depending on the conditions, the di- and triphenylalanine derivatives self-organize into a wide variety of stable polymorphic structures, which have been characterized: stacked braids, doughnut-like shapes, bundled arrays of nanotubes, corkscrew-like shapes and spherulitic microstructures. These highly aromatic Phe-based peptides also form incipient branched dendritic microstructures, even though they are highly unstable, making their manipulation very difficult. Conversely, the tetraphenylalanine derivative spontaneously self-assembles into stable dendritic microarchitectures made of branches growing from nucleated primary frameworks. The fractal dimension of these microstructures is â¼1.70, which provides evidence for self-similarity and two-dimensional diffusion controlled growth. DFT calculations at the M06L/6-31G(d) level have been carried out on model ß-sheets since this is the most elementary building block of Phe-based peptide polymorphs. The results indicate that the antiparallel ß-sheet is more stable than the parallel one, with the difference between them growing with the number of Phe residues. Thus, the cooperative effects associated with the antiparallel disposition become more favorable when the number of Phe residues increases from 2 to 4, while those of the parallel disposition remained practically constant.
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Péptidos/química , Fenilalanina/química , Nanotubos , Conformación ProteicaRESUMEN
Acute respiratory infections are responsible for high morbi-mortality in Peruvian children. However, the etiological agents are poorly identified. This study, conducted during the pandemic outbreak of H1N1 influenza in 2009, aims to determine the main etiological agents responsible for acute respiratory infections in children from Lima, Peru. Nasopharyngeal swabs collected from 717 children with acute respiratory infections between January 2009 and December 2010 were analyzed by multiplex RT-PCR for 13 respiratory viruses: influenza A, B, and C virus; parainfluenza virus (PIV) 1, 2, 3, and 4; and human respiratory syncytial virus (RSV) A and B, among others. Samples were also tested with direct fluorescent-antibodies (DFA) for six respiratory viruses. RT-PCR and DFA detected respiratory viruses in 240 (33.5%) and 85 (11.9%) cases, respectively. The most common etiological agents were RSV-A (15.3%), followed by influenza A (4.6%), PIV-1 (3.6%), and PIV-2 (1.8%). The viruses identified by DFA corresponded to RSV (5.9%) and influenza A (1.8%). Therefore, respiratory syncytial viruses (RSV) were found to be the most common etiology of acute respiratory infections. The authors suggest that active surveillance be conducted to identify the causative agents and improve clinical management, especially in the context of possible circulation of pandemic viruses.
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Virus de la Influenza A/aislamiento & purificación , Virus de la Parainfluenza 1 Humana/aislamiento & purificación , Virus de la Parainfluenza 2 Humana/aislamiento & purificación , Virus Sincitial Respiratorio Humano/aislamiento & purificación , Infecciones del Sistema Respiratorio/epidemiología , Infecciones del Sistema Respiratorio/virología , Enfermedad Aguda , Niño , Femenino , Técnica del Anticuerpo Fluorescente Directa , Humanos , Incidencia , Lactante , Subtipo H1N1 del Virus de la Influenza A/aislamiento & purificación , Gripe Humana/epidemiología , Masculino , Reacción en Cadena de la Polimerasa Multiplex , Nasofaringe/virología , Perú/epidemiología , Infecciones por Virus Sincitial Respiratorio/epidemiología , Infecciones por Respirovirus/epidemiología , Infecciones por Rubulavirus/epidemiología , Factores de TiempoRESUMEN
Although the content of Mg(2+) in hard tissues is very low (typically ≤1.5 wt %), its incorporation into synthetic hydroxyapatite (HAp) particles and its role in the mineral's properties are still subject of intensive debate. A combined experimental-computational approach is used to answer many of the open questions. Mg(2+) -enriched HAp particles are prepared using different synthetic approaches and considering different concentrations of Mg(2+) in the reaction medium. The composition, morphology and structure of the resulting particles are investigated using X-ray photoelectron spectroscopy, energy dispersive X-ray spectroscopy, scanning and transmission electron microscopies, FTIR, and wide-angle X-ray diffraction. After this scrutiny, the role of the Mg(2+) in the first nucleation stages, before HAp formation, is investigated using atomistic molecular dynamics simulations. Saturated solutions are simulated with and without the presence of DNA, which has been recently used as a soft template in the biomineralization process. This synergistic investigation provides a complete picture of how Mg(2+) ions affect the mineralization from the first stages onwards.