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1.
Animals (Basel) ; 13(24)2023 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-38136921

RESUMO

This study analyzed the effects of an Alliaceae encapsulated extract (AE-e) on daily gain (ADG), feed intake (ADFI), feed conversion ratio (FCR), oocysts per gram of feces (OPG), intestinal lesion (LS), and microbiota composition in broilers challenged with Eimeria spp. A total of 4800 one day Cobb-500 were allotted into 10 treatment groups with 12 replicates of 40 birds in a 2 × 4 + 2 factorial arrangement. The first factor was non-challenged (NC) or challenged (C), the second was four levels of AE-e added in the basal diet, 0 (AE0), 250 (AE250), 500 (AE500), and 750 mg·kg-1 (AE750), plus two ionophore controls, non-challenged (NC-Ion) and challenged (C-Ion). No interactions were observed between factors (NC0, NC250, NC500, NC750, C0, C250, C500, and C750), while C-Ion improved FCR at 21 d. The challenge affected negatively ADG and FCR and promoted enteropathogens in cecum. AE750 improved FCR in the finisher and cumulative phases, while C-Ion had fewer total OPG than C0 and C250. Likewise, at 21d, C250, C500, and C-Ion had fewer LS than C0, while at 28 d, C750 showed lower than C-Ion. In the cecum microbiota, C500 had more Ruminococcus, Firmicutes b, and Intestinimonas than C-Ion. In summary, AE-e showed beneficial results in broilers infected with Eimeria spp.

2.
Int J Mol Sci ; 24(19)2023 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-37834188

RESUMO

Photoproteins, luminescent proteins or optoproteins are a kind of light-response protein responsible for the conversion of light into biochemical energy that is used by some bacteria or fungi to regulate specific biological processes. Within these specific proteins, there are groups such as the photoreceptors that respond to a given light wavelength and generate reactions susceptible to being used for the development of high-novel applications, such as the optocontrol of metabolic pathways. Photoswitchable proteins play important roles during the development of new materials due to their capacity to change their conformational structure by providing/eliminating a specific light stimulus. Additionally, there are bioluminescent proteins that produce light during a heatless chemical reaction and are useful to be employed as biomarkers in several fields such as imaging, cell biology, disease tracking and pollutant detection. The classification of these optoproteins from bacteria and fungi as photoreceptors or photoresponse elements according to the excitation-emission spectrum (UV-Vis-IR), as well as their potential use in novel applications, is addressed in this article by providing a structured scheme for this broad area of knowledge.


Assuntos
Bactérias , Proteínas Luminescentes/metabolismo , Bactérias/metabolismo
3.
Micromachines (Basel) ; 14(10)2023 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-37893319

RESUMO

Copper nanoparticles (CuNPs) can be synthesized by green methods using plant extracts. These methods are more environmentally friendly and offer improved properties of the synthesized NPs in terms of biocompatibility and functional capabilities. Traditional medicine has a rich history of utilization of herbs for millennia, offering a viable alternative or complementary option to conventional pharmacological medications. Plants of traditional herbal use or those with medicinal properties are candidates to be used to obtain NPs due to their high and complex content of biocompounds with different redox capacities that provide a dynamic reaction environment for NP synthesis. Other synthesis conditions, such as salt precursor concentration, temperature, time synthesis, and pH, have a significant effect on the characteristics of the NPs. This paper will review the properties of some compounds from medicinal plants, plant extract obtention methods alternatives, characteristics of plant extracts, and how they relate to the NP synthesis process. Additionally, the document includes diverse applications associated with CuNPs, starting from antibacterial properties to potential applications in metabolic disease treatment, vegetable tissue culture, therapy, and cardioprotective effect, among others.

4.
Clin. transl. oncol. (Print) ; 25(4): 882-896, abr. 2023.
Artigo em Inglês | IBECS | ID: ibc-217749

RESUMO

Oligometastatic disease (OMD) defines a cancer status that is intermediate between localized and widely spread metastatic disease, and can be treated with curative intent. While diagnostic imaging tools have considerably improved in recent years, unidentified micrometastases can still evade current detection techniques, allowing the disease to progress. The various OMD scenarios are mainly defined by the number of metastases, the biological and molecular tumour profiles, and the timing of the development of metastases. Increasing knowledge has contributed to the earlier and improved detection of OMD, underlining the importance of early disease control. In view of increasing OMD detection rates in current real-world clinical practice and the lack of standardized evidence-based guidelines to treat this cancer status, a board of experts from the Spanish Societies of Radiation Oncology (SEOR) and Medical Oncology (SEOM) organized a series of sessions to update the current state-of-the-art on OMD from a multidisciplinary perspective, and to discuss how results from clinical studies might translate into promising treatment options. This expert review series summarizes what is known and what it is pending clarification in the context of OMD in the scenarios of non-small cell lung cancer and breast cancer (Part I), and prostate cancer and colorectal cancer (Part II), aiming to offer specialists a pragmatic framework to help improve patient management (AU)


Assuntos
Humanos , Neoplasias da Mama/terapia , Carcinoma Pulmonar de Células não Pequenas/terapia , Neoplasias Pulmonares/terapia , Carcinoma Pulmonar de Células não Pequenas/patologia , Neoplasias Pulmonares/patologia , Neoplasias da Mama/patologia , Radiocirurgia/métodos
5.
Clin. transl. oncol. (Print) ; 25(4): 897-911, abr. 2023.
Artigo em Inglês | IBECS | ID: ibc-217750

RESUMO

Oligometastatic disease (OMD) defines a status of cancer that is intermediate between localized and widely spread metastatic disease, and can be treated with curative intent. While imaging diagnostic tools have considerably improved in recent years, unidentified micrometastases can still escape from current detection techniques allowing disease to progress. The variety of OMD scenarios are mainly defined by the number of metastases, the biological and molecular tumour profiles, and the timing of the development of metastases. Increasing knowledge has contributed to the earlier and improved detection of OMD, underlining the importance of an early disease control. Based on increasing detection rates of OMD in the current real clinical practice and the lack of standardized evidence-based guidelines to treat this cancer status, a board of experts from the Spanish Societies of Radiation Oncology (SEOR) and Medical Oncology (SEOM) organized a series of sessions to update the current state-of-the-art on OMD from a multidisciplinary perspective, and to discuss how results from clinical studies may translate into promising treatment options. This experts’ review series summarizes what is known and what it is pending clarification in the context of OMD in the scenarios of Non-Small Cell Lung Cancer and Breast Cancer (Part I), and Prostate Cancer and Colorectal Cancer (Part II), aiming to offer specialists a pragmatic framework that might contribute to the improved management of patients (AU)


Assuntos
Humanos , Masculino , Neoplasias Colorretais/radioterapia , Neoplasias da Próstata/radioterapia , Neoplasias Colorretais/cirurgia , Neoplasias da Próstata/cirurgia , Radiocirurgia/métodos , Neoplasias Colorretais/patologia , Neoplasias da Próstata/patologia
6.
Animals (Basel) ; 13(6)2023 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-36978663

RESUMO

This study analyzed the effects of different dietary doses of encapsulated propyl propane thiosulfonate (Pe-PTSO) on the apparent ileal digestibility (AID) of nutrients and productive performance in broilers. A total of 100 one-day-old Cobb 500 were housed in battery cages for 20 days. At 10 days of age, the birds were assigned to one of five diets: negative control (P0), 250 mg/kg of Pe-PTSO (P250), 500 mg/kg of Pe-PTSO (P500), 750 mg/kg of Pe-PTSO (P750), and positive control, nicarbazin-narasin (ION). Titanium dioxide was the external marker, which was added to the diets from day 17 to 20. In the birds fed the P250 diet, there was a significant difference (p ≤ 0.05) in the AID values for amino acids and energy compared to those that consumed the P0 diet. Furthermore, the P250 diet significantly increased (p ≤ 0.05) the average daily weight gain compared to the P0 diet. No significant differences were observed between treatments in average daily feed intake and feed conversion ratio. In summary, the inclusion of 250 mg of encapsulated PTSO per kg in broiler chickens diet improved the digestibility of amino acids and energy, as well as weight gain.

7.
Nanoscale Adv ; 5(6): 1611-1623, 2023 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-36926558

RESUMO

Oligopeptide end-modified poly(ß-amino ester)s (OM-pBAEs) offer a means for the effective implementation of gene therapeutics in the near future. A fine-tuning of OM-pBAEs to meet application requirements is achieved by the proportional balance of oligopeptides used and provide gene carriers with high transfection efficacy, low toxicity, precise targeting, biocompatibility, and biodegradability. Understanding the influence and conformation of each building block at molecular and biological levels is therefore pivotal for further development and improvement of these gene carriers. Herein, we unmask the role of individual OM-pBAE components and their conformation in OM-pBAE/polynucleotide nanoparticles using a combination of fluorescence resonance energy transfer, enhanced darkfield spectral microscopy, atomic force microscopy, and microscale thermophoresis. We found that modifying the pBAE backbone with three end-terminal amino acids produces unique mechanical and physical properties for each combination. Higher adhesion properties are seen with arginine and lysine-based hybrid nanoparticles, while histidine provides an advantage in terms of construct stability. Our results shed light on the high potential of OM-pBAEs as gene delivery vehicles and provide insights into the influence of the nature of surface charges and the chemical nature of the pBAE modifications on their paths towards endocytosis, endosomal escape, and transfection.

8.
Colloids Surf B Biointerfaces ; 222: 113019, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36435028

RESUMO

Drug and gene delivery systems based on polymeric nanoparticles offer a greater efficacy and a reduced toxicity compared to traditional formulations. Recent studies have evidenced that their internalization, biodistribution and efficacy can be affected, among other factors, by their mechanical properties. Here, we analyze by means of Atomic Force Microscopy force spectroscopy how composition, surface functionalization and loading affect the mechanics of nanoparticles. For this purpose, nanoparticles made of Poly(lactic-co-glycolic) (PLGA) and Ethyl cellulose (EC) with different functionalizations and loading were prepared by nano-emulsion templating using the Phase Inversion Composition method (PIC) to form the nano-emulsions. A multiparametric nanomechanical study involving the determination of the Young's modulus, maximum deformation and breakthrough force was carried out. The obtained results showed that composition, surface functionalization and loading affect the nanomechanical properties in a different way, thus requiring, in general, to consider the overall mechanical properties after the addition of a functionalization or loading. A graphical representation method has been proposed enabling to easily identify mechanically equivalent formulations, which is expected to be useful in the development of soft polymeric nanoparticles for pre-clinical and clinical use.


Assuntos
Nanopartículas , Ácido Poliglicólico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Ácido Poliglicólico/química , Ácido Láctico/química , Distribuição Tecidual , Nanopartículas/química
9.
Clin Transl Oncol ; 25(4): 897-911, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36525230

RESUMO

Oligometastatic disease (OMD) defines a status of cancer that is intermediate between localized and widely spread metastatic disease, and can be treated with curative intent. While imaging diagnostic tools have considerably improved in recent years, unidentified micrometastases can still escape from current detection techniques allowing disease to progress. The variety of OMD scenarios are mainly defined by the number of metastases, the biological and molecular tumour profiles, and the timing of the development of metastases. Increasing knowledge has contributed to the earlier and improved detection of OMD, underlining the importance of an early disease control. Based on increasing detection rates of OMD in the current real clinical practice and the lack of standardized evidence-based guidelines to treat this cancer status, a board of experts from the Spanish Societies of Radiation Oncology (SEOR) and Medical Oncology (SEOM) organized a series of sessions to update the current state-of-the-art on OMD from a multidisciplinary perspective, and to discuss how results from clinical studies may translate into promising treatment options. This experts' review series summarizes what is known and what it is pending clarification in the context of OMD in the scenarios of Non-Small Cell Lung Cancer and Breast Cancer (Part I), and Prostate Cancer and Colorectal Cancer (Part II), aiming to offer specialists a pragmatic framework that might contribute to the improved management of patients.


Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Colorretais , Neoplasias Pulmonares , Neoplasias da Próstata , Radiocirurgia , Masculino , Humanos , Carcinoma Pulmonar de Células não Pequenas/patologia , Neoplasias Pulmonares/patologia , Neoplasias da Próstata/radioterapia , Neoplasias da Próstata/patologia , Oncologia , Neoplasias Colorretais/terapia , Neoplasias Colorretais/patologia , Radiocirurgia/métodos
10.
Clin Transl Oncol ; 25(4): 882-896, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36525231

RESUMO

Oligometastatic disease (OMD) defines a cancer status that is intermediate between localized and widely spread metastatic disease, and can be treated with curative intent. While diagnostic imaging tools have considerably improved in recent years, unidentified micrometastases can still evade current detection techniques, allowing the disease to progress. The various OMD scenarios are mainly defined by the number of metastases, the biological and molecular tumour profiles, and the timing of the development of metastases. Increasing knowledge has contributed to the earlier and improved detection of OMD, underlining the importance of early disease control. In view of increasing OMD detection rates in current real-world clinical practice and the lack of standardized evidence-based guidelines to treat this cancer status, a board of experts from the Spanish Societies of Radiation Oncology (SEOR) and Medical Oncology (SEOM) organized a series of sessions to update the current state-of-the-art on OMD from a multidisciplinary perspective, and to discuss how results from clinical studies might translate into promising treatment options. This expert review series summarizes what is known and what it is pending clarification in the context of OMD in the scenarios of non-small cell lung cancer and breast cancer (Part I), and prostate cancer and colorectal cancer (Part II), aiming to offer specialists a pragmatic framework to help improve patient management.


Assuntos
Neoplasias da Mama , Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Neoplasias da Próstata , Radiocirurgia , Masculino , Humanos , Carcinoma Pulmonar de Células não Pequenas/radioterapia , Carcinoma Pulmonar de Células não Pequenas/patologia , Neoplasias da Mama/terapia , Neoplasias Pulmonares/radioterapia , Neoplasias Pulmonares/patologia , Oncologia , Radiocirurgia/métodos
11.
Biosensors (Basel) ; 12(11)2022 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-36354471

RESUMO

Metamaterials are broadly defined as artificial, electromagnetically homogeneous structures that exhibit unusual physical properties that are not present in nature. They possess extraordinary capabilities to bend electromagnetic waves. Their size, shape and composition can be engineered to modify their characteristics, such as iridescence, color shift, absorbance at different wavelengths, etc., and harness them as biosensors. Metamaterial construction from biological sources such as carbohydrates, proteins and nucleic acids represents a low-cost alternative, rendering high quantities and yields. In addition, the malleability of these biomaterials makes it possible to fabricate an endless number of structured materials such as composited nanoparticles, biofilms, nanofibers, quantum dots, and many others, with very specific, invaluable and tremendously useful optical characteristics. The intrinsic characteristics observed in biomaterials make them suitable for biomedical applications. This review addresses the optical characteristics of metamaterials obtained from the major macromolecules found in nature: carbohydrates, proteins and DNA, highlighting their biosensor field use, and pointing out their physical properties and production paths.


Assuntos
Técnicas Biossensoriais , Nanopartículas , Materiais Biocompatíveis , DNA , Carboidratos
12.
Indoor Air ; 32(10): e13118, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36305066

RESUMO

SARS-CoV-2 has been detected both in air and on surfaces, but questions remain about the patient-specific and environmental factors affecting virus transmission. Additionally, more detailed information on viral sampling of the air is needed. This prospective cohort study (N = 56) presents results from 258 air and 252 surface samples from the surroundings of 23 hospitalized and eight home-treated COVID-19 index patients between July 2020 and March 2021 and compares the results between the measured environments and patient factors. Additionally, epidemiological and experimental investigations were performed. The proportions of qRT-PCR-positive air (10.7% hospital/17.6% homes) and surface samples (8.8%/12.9%) showed statistical similarity in hospital and homes. Significant SARS-CoV-2 air contamination was observed in a large (655.25 m3 ) mechanically ventilated (1.67 air changes per hour, 32.4-421 L/s/patient) patient hall even with only two patients present. All positive air samples were obtained in the absence of aerosol-generating procedures. In four cases, positive environmental samples were detected after the patients had developed a neutralizing IgG response. SARS-CoV-2 RNA was detected in the following particle sizes: 0.65-4.7 µm, 7.0-12.0 µm, >10 µm, and <100 µm. Appropriate infection control against airborne and surface transmission routes is needed in both environments, even after antibody production has begun.


Assuntos
Poluição do Ar em Ambientes Fechados , COVID-19 , Humanos , SARS-CoV-2 , COVID-19/epidemiologia , RNA Viral , Estudos Prospectivos , Aerossóis e Gotículas Respiratórios
13.
Physiol Mol Biol Plants ; 27(8): 1651-1657, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34539108

RESUMO

Pepper seeds are slow to germinate and emergence is often non-uniform and incomplete, reducing gains from this cash crop. This study investigated the effects of pineapple stem- derived protease (stem bromelain) based priming on pepper seed germination in relation to reserve mobilization (specifically, proteins and amino acids), germination, emergence and plant growth. These parameters were compared across two controls, (1) unsoaked seeds and (2) seeds soaked in deionized water, and seeds soaked in pineapple stem bromelain crude extract (treatment). Seeds were soaked in bromelain crude extract possessing a proteolytic activity of 6.25 tU or deionized water (first control) for 3 h at 35 °C. Light microscopy revealed an abundance of protein bodies in the endosperm of the seeds prior to imbibition. When observed for a period of 96 h, these bodies were progressively degraded, with the rate of this degradation being fastest in bromelain-treated seeds. Quantitative analysis of protein levels confirmed this observation: 17.2 mg proteins/g FW at 120 h after priming in bromelain-treated seeds compared with 22.1 mg/g FW in controls (average). The bromelain treatment also increased levels of free amino acids from 3.9 mg/g FW in the controls to 4.6 mg/g FW after 120 h of imbibition. Germination and emergence percentages were initially higher in bromelain-treated seeds: 92.0% germination in bromelain-treated seeds vs. ~ 52.2% in the controls at 18 d; 100% emergence in protease-treated seeds vs. ~ 72.2% in the controls at 18 d. However, these parameters were comparable across the treatment and the controls at 28 d. Importantly, plant fresh and dry weights were significantly higher when seeds were primed with bromelain. The results suggest the use of bromelain extracts for priming pepper seeds based on their proteolytic activity, since germination is dependent on the availability of crude protein and essential amino acids. The benefits of bromelain seed priming appear to translate into improved seedling growth as well.

14.
J Nanobiotechnology ; 19(1): 167, 2021 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-34082783

RESUMO

Liposomes are widely used as drug delivery carriers and as cell model systems. Here, we measure the dielectric properties of individual liposomes adsorbed on a metal electrode by in-liquid scanning dielectric microscopy in force detection mode. From the measurements the lamellarity of the liposomes, the separation between the lamellae and the specific capacitance of the lipid bilayer can be obtained. As application we considered the case of non-extruded DOPC liposomes with radii in the range ~ 100-800 nm. Uni-, bi- and tri-lamellar liposomes have been identified, with the largest population corresponding to bi-lamellar liposomes. The interlamellar separation in the bi-lamellar liposomes is found to be below ~ 10 nm in most instances. The specific capacitance of the DOPC lipid bilayer is found to be ~ 0.75 µF/cm2 in excellent agreement with the value determined on solid supported planar lipid bilayers. The lamellarity of the DOPC liposomes shows the usual correlation with the liposome's size. No correlation is found, instead, with the shape of the adsorbed liposomes. The proposed approach offers a powerful label-free and non-invasive method to determine the lamellarity and dielectric properties of single liposomes.


Assuntos
Portadores de Fármacos , Lipossomos/química , Microscopia , Sistemas de Liberação de Medicamentos , Bicamadas Lipídicas , Nanotecnologia/métodos
15.
Viruses ; 13(4)2021 03 31.
Artigo em Inglês | MEDLINE | ID: mdl-33807151

RESUMO

The renal involvement of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been reported. The etiology of kidney injury appears to be tubular, mainly due to the expression of angiotensin-converting enzyme 2, the key joint receptor for SARS-CoV-2; however, cases with glomerular implication have also been documented. The multifactorial origin of this renal involvement could include virus-mediated injury, cytokine storm, angiotensin II pathway activation, complement dysregulation, hyper-coagulation, and microangiopathy. We present the renal histological findings from a patient who developed acute kidney injury and de novo nephrotic syndrome, highly suggestive of acute IgA-dominant infection-associated glomerulonephritis (IgA-DIAGN) after SARS-CoV-2 infection, as evidenced by the presence of this virus detected in the renal tissue of the patient via immunohistochemistry assay. In summary, we document the first case of IgA-DIAGN associated to SARS-CoV-2. Thus, SARS-CoV-2 S may act as a super antigen driving the development of multisystem inflammatory syndrome as well as cytokine storm in patients affected by COVID-19, reaching the glomerulus and leading to the development of this novel IgA-DIAGN.


Assuntos
COVID-19/complicações , Glomerulonefrite/etiologia , Glomerulonefrite/imunologia , Imunoglobulina A/imunologia , Idoso de 80 Anos ou mais , COVID-19/virologia , Síndrome da Liberação de Citocina , Humanos , Rim/imunologia , Masculino , SARS-CoV-2/fisiologia
16.
Eur J Pharm Biopharm ; 156: 155-164, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32927077

RESUMO

During the last few decades, extensive efforts has been made to design nanocarriers to transport drugs into the central nervous system (CNS). However, its efficacy is limited due to the presence of the Blood-Brain Barrier (BBB) which greatly reduces drug penetration making Drug Delivery Systems (DDS) necessary. Polymeric nanoparticles (NPs) have been reported to be appropriate for this purpose and in particular, poly(lactic-co-glycolic acid) (PLGA) has been used for its ability to entrap small molecule drugs with great efficiency and the ease with which it functionalizes NPs. Despite the fact that their synthetic identity has been studied in depth, the biological identity of such manufactured polymers still remains unknown as does their biodistribution and in vivo fate. This biological identity is a result of their interaction with blood proteins, the so-called "protein corona" which tends to alter the behavior of polymeric nanoparticles in the body. The aim of the present research is to identify the proteins bounded to polymeric nanoparticles designed to selectively interact with the BBB. For this purpose, four different PLGA NPs were prepared and analyzed: (i) "PLGA@Drug," in which a model drug was encapsulated in its core; (ii) "8D3-PLGA" NPs where the PLGA surface was functionalized with a monoclonal anti-transferrin receptor antibody (8D3 mAb) in order to specifically target the BBB; (iii) "8D3-PLGA@Drug" in which the PLGA@Drug surface was functionalized using the same antibody described above and (iv) bare PLGA NPs which were used as a control. Once the anticipated protein corona NPs were obtained, proteins decorating both bare and functionalized PLGA NPs were isolated and analyzed. Apart from the indistinct interaction with PLGA NPs with the most abundant serum proteins, specific proteins could also be identified in the case of functionalized PLGA NPs. These findings may provide valuable insight into designing novel vehicles based on PLGA NPs for crossing the BBB.


Assuntos
Barreira Hematoencefálica/metabolismo , Nanopartículas/metabolismo , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/metabolismo , Proteínas/metabolismo , Tiazolidinedionas/metabolismo , Barreira Hematoencefálica/efeitos dos fármacos , Emulsões/síntese química , Emulsões/metabolismo , Humanos , Nanopartículas/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/síntese química , Transporte Proteico/efeitos dos fármacos , Transporte Proteico/fisiologia , Proteínas/síntese química , Tiazolidinedionas/síntese química
17.
ACS Appl Mater Interfaces ; 11(37): 33620-33627, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31448892

RESUMO

Cell lipid membranes are the site of vital biological processes, such as motility, trafficking, and sensing, many of which involve mechanical forces. Elucidating the interplay between such bioprocesses and mechanical forces requires the use of tools that apply and measure piconewton-level forces, e.g., optical tweezers. Here, we introduce the combination of optical tweezers with free-standing lipid bilayers, which are fully accessible on both sides of the membrane. In the vicinity of the lipid bilayer, optical trapping would normally be impossible due to optical distortions caused by pockets of the solvent trapped within the membrane. We solve this by drastically reducing the size of these pockets via tuning of the solvent and flow cell material. In the resulting flow cells, lipid nanotubes are straightforwardly pushed or pulled and reach lengths above half a millimeter. Moreover, the controlled pushing of a lipid nanotube with an optically trapped bead provides an accurate and direct measurement of important mechanical properties. In particular, we measure the membrane tension of a free-standing membrane composed of a mixture of dioleoylphosphatidylcholine (DOPC) and dipalmitoylphosphatidylcholine (DPPC) to be 4.6 × 10-6 N/m. We demonstrate the potential of the platform for biophysical studies by inserting the cell-penetrating trans-activator of transcription (TAT) peptide in the lipid membrane. The interactions between the TAT peptide and the membrane are found to decrease the value of the membrane tension to 2.1 × 10-6 N/m. This method is also fully compatible with electrophysiological measurements and presents new possibilities for the study of membrane mechanics and the creation of artificial lipid tube networks of great importance in intra- and intercellular communication.


Assuntos
Membrana Celular/química , Dispositivos Lab-On-A-Chip , Bicamadas Lipídicas/química , Nanotubos/química , Pinças Ópticas , 1,2-Dipalmitoilfosfatidilcolina/química , Fosfatidilcolinas/química , Tensão Superficial
18.
Colloids Surf B Biointerfaces ; 172: 400-406, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30195157

RESUMO

In this study, we show that dry saturated phospholipid layers prepared by the spin-coating technique could present thinner regions associated to interdigitated phases under some conditions. The morphological characteristics of lipid layers of saturated phosphocholines, such as dilauroylphosphatidylcholine (DLPC), dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC) and distearoylphosphatidylcholine (DSPC), have been measured by Atomic Force Microscopy and revealed that the presence of interdigitated regions is not induced by the same parameters that induce them in hydrated samples. To achieve these results the effect of the lipid hidrocabonated chain length, the presence of alcohol in the coating solution, the spinning velocity and the presence of cholesterol were tested. We showed that DPPC and DSPC bilayers, on the one side, can show structures with similar height than interdigitated regions observed in hydrated samples, while, on the other side, DLPC and DMPC tend to show no evidence of interdigitation. Results indicate that the presence of interdigitated areas is due to the presence of lateral tensions and, hence, that they can be eliminated by releasing these tensions by, for instance, the addition of cholesterol. These results demonstrate that interdigitation in lipid layers is a rather general phenomena and can be observed in lipid bilayers in dry conditions.


Assuntos
Ar , Bicamadas Lipídicas/química , Álcoois/química , Colesterol/química , Lipídeos/química , Microscopia de Força Atômica , Soluções
19.
Pharm Res ; 34(5): 1093-1103, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-28197757

RESUMO

PURPOSE: Gold nanoparticles have been proved useful for many biomedical applications, specifically, for their use as advanced imaging systems. However, they usually present problems related with stability and toxicity. METHODS: In the present work, gold-nanoparticles have been encapsulated in polymeric nanoparticles using a novel methodology based on nano-emulsion templating. Firstly, gold nanoparticles have been transferred from water to ethyl acetate, a solvent classified as class III by the NIH guidelines (low toxic potential). Next, the formation of nano-emulsions loaded with gold nanoparticles has been performed using a low-energy, the phase inversion composition (PIC) emulsification method, followed by solvent evaporation giving rise to polymeric nanoparticles. RESULTS: Using this methodology, high concentrations of gold nanoparticles (>100 pM) have been encapsulated. Increasing gold nanoparticle concentration, nano-emulsion and nanoparticle sizes increase, resulting in a decrease on the stability. It is noteworthy that the designed nanoparticles did not produce cytotoxicity neither hemolysis at the required concentration. CONCLUSIONS: Therefore, it can be concluded that a novel and very versatile methodology has been developed for the production of polymeric nanoparticles loaded with gold nanoparticles. Graphical Abstract Schematic representation of AuNP-loaded polymeric nanoparticles preparation from nano-emulsion templating.


Assuntos
Emulsões/química , Ouro/química , Ácido Láctico/química , Nanopartículas Metálicas/química , Ácido Poliglicólico/química , Acetatos/química , Linhagem Celular Tumoral , Portadores de Fármacos/química , Células HeLa , Humanos , Tamanho da Partícula , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Polímeros/química , Solventes/química , Água/química
20.
Artif Intell Med ; 70: 12-30, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27431034

RESUMO

INTRODUCTION: Numeric time series are present in a very wide range of domains, including many branches of medicine. Data mining techniques have proved to be useful for knowledge discovery in this type of data and for supporting decision-making processes. OBJECTIVES: The overall objective is to classify time series based on the discovery of frequent patterns. These patterns will be discovered in symbolic sequences obtained from the time series data by means of a temporal abstraction process. METHODS: Firstly, we transform numeric time series into symbolic time sequences, where the symbols aim to represent the relevant domain concepts. These symbols can be defined using either public or expert domain knowledge. Then we apply a symbolic pattern discovery technique to the output symbolic sequences. This technique identifies the subsequences frequently found in a population group. These subsequences (patterns) are representative of population groups. Finally, we employ a classification technique based on the identified patterns in order to classify new individuals. Thanks to the inclusion of domain knowledge, the classification results can be explained using domain terminology. This makes the results easier to interpret for the domain specialist (physician). RESULTS: This method has been applied to brainstem auditory evoked potentials (BAEPs) time series. Preliminary experiments were carried out to analyse several aspects of the method including the best configuration of the pattern discovery technique parameters. We then applied the method to the BAEPs of 83 individuals belonging to four classes (healthy, conductive hearing loss, vestibular schwannoma-brainstem involvement and vestibular schwannoma-8th-nerve involvement). According to the results of the cross-validation, overall accuracy was 99.4%, sensitivity (recall) was 97.6% and specificity was 100% (no false positives). CONCLUSION: The proposed method effectively reduces dimensionality. Additionally, if the symbolic transformation includes the right domain knowledge, the method arguably outputs a data representation that denotes the relevant domain concepts more clearly. The method is capable of finding patterns in BAEPs time series and is very accurate at correctly predicting whether or not new patients have an auditory-related disorder.


Assuntos
Mineração de Dados , Potenciais Evocados Auditivos do Tronco Encefálico , Humanos , Reconhecimento Automatizado de Padrão , Sensibilidade e Especificidade
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