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1.
Int J Implant Dent ; 10(1): 11, 2024 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-38472687

RESUMO

OBJECTIVE: This study analyzed and compared the biomechanical properties of maxillary sinus floor mucosa with implants at three different maxillary sinus angles during a modified internal sinus floor elevation procedure. METHODS: 3D reconstruction of the implant, maxillary sinus bone, and membrane were performed. The maxillary sinus model was set at three different angles. Two internal maxillary sinus elevation models were established, and finite element analysis was used to simulate the modified maxillary sinus elevation process. The implant was elevated to 10 mm at three maxillary sinus angles when the maxillary sinus floor membrane was separated by 0 and 4 mm. The stress of the maxillary sinus floor membrane was analyzed and compared. RESULTS: When the maxillary sinus floor membrane was separated by 0 mm and elevated to 10 mm, the peak stress values of the implant on the maxillary sinus floor membrane at three different angles were as follows: maxillary sinus I: 5.14-78.32 MPa; maxillary sinus II: 2.81-73.89 MPa; and maxillary sinus III: 2.82-51.87 MPa. When the maxillary sinus floor membrane was separated by 4 mm and elevated to 10 mm, the corresponding values were as follows: maxillary sinus I: 0.50-7.25 MPa; maxillary sinus II: 0.81-16.55 MPa; and maxillary sinus III: 0.49-22.74 MPa. CONCLUSION: The risk of sinus floor membrane rupture is greatly reduced after adequate dissection of the maxillary sinus floor membrane when performing modified internal sinus elevation in a narrow maxillary sinus. In a wide maxillary sinus, the risk of rupture or perforation of the wider maxillary sinus floor is reduced, regardless of whether traditional or modified internal sinus elevation is performed at the same height.


Assuntos
Implantes Dentários , Levantamento do Assoalho do Seio Maxilar , Implantação Dentária Endóssea/métodos , Levantamento do Assoalho do Seio Maxilar/métodos , Seio Maxilar/cirurgia , Membranas/cirurgia
2.
Int J Mol Sci ; 25(5)2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-38474151

RESUMO

Extracellular vesicles (EVs) are lipid bilayers derived from cell membranes, released by both eukaryotic cells and bacteria into the extracellular environment. During production, EVs carry proteins, nucleic acids, and various compounds, which are then released. While Gram-positive bacteria were traditionally thought incapable of producing EVs due to their thick peptidoglycan cell walls, recent studies on membrane vesicles (MVs) in Gram-positive bacteria have revealed their significant role in bacterial physiology and disease progression. This review explores the current understanding of MVs in Gram-positive bacteria, including the characterization of their content and functions, as well as their interactions with host and bacterial cells. It offers a fresh perspective to enhance our comprehension of Gram-positive bacterial EVs.


Assuntos
Vesículas Extracelulares , Bactérias Gram-Positivas , Bactérias , Membranas , Membrana Celular , Bicamadas Lipídicas/metabolismo , Vesículas Extracelulares/metabolismo
3.
J Vis Exp ; (204)2024 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-38465944

RESUMO

The precise localization and activation of proteins at the cell membrane at a certain time gives rise to many cellular processes, including cell polarization, migration, and division. Thus, methods to recruit proteins to model membranes with subcellular resolution and high temporal control are essential when reproducing and controlling such processes in synthetic cells. Here, a method is described for fabricating light-regulated reversible protein patterns at lipid membranes with high spatiotemporal precision. For this purpose, we immobilize the photoswitchable protein iLID (improved light-inducible dimer) on supported lipid bilayers (SLBs) and on the outer membrane of giant unilamellar vesicles (GUVs). Upon local blue light illumination, iLID binds to its partner Nano (wild-type SspB) and allows the recruitment of any protein of interest (POI) fused to Nano from the solution to the illuminated area on the membrane. This binding is reversible in the dark, which provides dynamic binding and release of the POI. Overall, this is a flexible and versatile method for regulating the localization of proteins with high precision in space and time using blue light.


Assuntos
Bicamadas Lipídicas , Proteínas , Proteínas/metabolismo , Membrana Celular/metabolismo , Bicamadas Lipídicas/metabolismo , Membranas , Lipossomas Unilamelares/metabolismo
4.
Methods Mol Biol ; 2774: 43-58, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38441757

RESUMO

Intercellular membrane-membrane interfaces are compartments with specialized functions and unique biophysical properties that are essential in numerous cellular processes including cell signaling, development, and immunity. Using synthetic biology to engineer or to create novel cellular functions in the intercellular regions has led to an increasing need for a platform that allows generation of functionalized intercellular membrane-membrane interfaces. Here, we present a synthetic biology platform to engineer functional membrane-membrane interfaces using a pair of dimerizing proteins in both cell-free and cellular environments. We envisage this platform to be a helpful tool for synthetic biologists who wish to engineer novel intercellular signaling and communication systems.


Assuntos
Transdução de Sinais , Biologia Sintética , Animais , Membranas , Biofísica , Dimerização , Mamíferos
5.
J Phys Chem B ; 128(11): 2734-2744, 2024 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-38459942

RESUMO

Voltage measurement via small-molecule fluorescent indicators is a valuable approach in deciphering complex dynamics in electrically excitable cells. However, our understanding of various physicochemical properties governing the performance of fluorescent voltage sensors based on the photoinduced electron transfer (PeT) mechanism remains incomplete. Here, through extensive molecular dynamics and free energy calculations, we systematically examine the orientation and membrane partition of three PeT-based voltage-sensing VoltageFluor (VF) dyes in different lipid environment. We show that the symmetry of the molecular scaffold and the net charge of the hydrophilic headgroup of a given VF dye dominate its orientation and membrane partition, respectively. Our work provides a mechanistic understanding of the physical properties contributing to the voltage sensitivity, signal-to-noise ratio, as well as membrane distribution of VF dyes and sheds light onto rational design principles of PeT-based fluorescent probes in general.


Assuntos
Corantes Fluorescentes , Simulação de Dinâmica Molecular , Corantes Fluorescentes/química , Potenciais da Membrana , Transporte de Elétrons , Membranas
6.
J Phys Chem B ; 128(11): 2632-2639, 2024 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-38467492

RESUMO

The cellular endocytosis of nanoparticles (NPs) is a fundamental biological process with significant potential in biomedical applications. However, a comprehensive understanding of the mechanistic aspects of endocytosis and the impact of particle properties on this process remains elusive. In this study, we investigated the membrane-wrapping behavior of soft NPs (SNPs) with varying rigidities using theoretical calculations. Our findings reveal that the membrane-wrapping process of SNPs involves a complex energy change including the possible existence of an energy barrier; moreover, it is found that the location and height of this barrier strongly depend on the mechanistic properties of the NPs and membranes. Additionally, by considering force balance in the membrane-wrapping process, we calculated the speed at which NP is internalized by the membrane, showing a nonmonotonic dependence on particle rigidity and/or wrapping degree. These phenomena can be attributed to competition between different energy components associated with NP-membrane binding, membrane tension, and deformations occurring during SNP-membrane interaction processes. Our results contribute to a deeper understanding of cellular-level endocytosis mechanisms and offer potential applications for soft NPs in biomedicine.


Assuntos
Nanopartículas , Membrana Celular/química , Nanopartículas/química , Membranas , Endocitose , Fenômenos Físicos
7.
Methods Mol Biol ; 2776: 185-196, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38502505

RESUMO

Diatoms such as Phaeodactylum tricornutum arose through a process termed secondary endosymbiosis, in which red alga-derived plastids are surrounded by a complicated membrane system. Subcellular marker proteins provide defined localizations on the compartmental and even sub-compartmental levels in the complex plastids of diatoms. Here we introduce how to use subcellular marker proteins and in vivo co-localization in the diatom P. tricornutum by presenting a step-by-step method allowing the determination of subcellular localization of proteins in different membranes of the secondary plastid. This chapter describes the materials required and the procedures of transformation and microscopic observation.


Assuntos
Diatomáceas , Diatomáceas/metabolismo , Proteínas/metabolismo , Membranas , Simbiose , Plastídeos/metabolismo
8.
Proc Natl Acad Sci U S A ; 121(10): e2319491121, 2024 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-38427601

RESUMO

Translocation of cytoplasmic molecules to the plasma membrane is commonplace in cell signaling. Membrane localization has been hypothesized to increase intermolecular association rates; however, it has also been argued that association should be faster in the cytosol because membrane diffusion is slow. Here, we directly compare an identical association reaction, the binding of complementary DNA strands, in solution and on supported membranes. The measured rate constants show that for a 10-µm-radius spherical cell, association is 22- to 33-fold faster at the membrane than in the cytoplasm. The kinetic advantage depends on cell size and is essentially negligible for typical ~1 µm prokaryotic cells. The rate enhancement is attributable to a combination of higher encounter rates in two dimensions and a higher reaction probability per encounter.


Assuntos
Transdução de Sinais , Citoplasma/metabolismo , Membrana Celular/metabolismo , Citosol/metabolismo , Membranas , Cinética
9.
Int J Oral Maxillofac Implants ; 39(1): 107-118, 2024 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-38416004

RESUMO

PURPOSE: To evaluate the impact of different approaches to sinus membrane perforation (SMP) repair on bone formation, postoperative complications, and implant loss risk. MATERIALS AND METHODS: Electronic searches on PubMed, Web of Science, Scopus, Embase, and Cochrane Library databases were conducted for publications up to February 2021. All included articles reported SMPs submitted for repair. The proportion of implant loss in repaired SMP sites was calculated using a random-effects model meta-analysis. RESULTS: A total of 130 studies reporting SMP repair protocols were included in the systematic review, with 20 selected for meta-analysis. A total of 1,972 sinuses that were perforated and repaired during sinus elevation using different approaches were included in the qualitative analysis. The resorbable collagen membrane was the most commonly reported treatment. The presence of sinusitis was the most frequently described complication. Regarding bone parameters, the majority of studies described no differences between perforated and repaired sinuses and intact membranes. No difference in the implant loss proportion was observed between sites with repaired SMP compared to undetected SMP. The proportion of implant loss in repaired sinuses membrane sites (independent of the material or implant placement time) was 4% (95% CI: 2.0 to 8.0). In meta-regression analysis, no association was observed between the SMP size and implant loss proportion (P = .86). CONCLUSIONS: The materials and techniques indicated for SMP management seem to securely seal the maxillary sinus, without a negative effect on the ultimate survival of the implants placed in the affected sinuses.


Assuntos
Seio Maxilar , Complicações Pós-Operatórias , Humanos , Seio Maxilar/cirurgia , Membranas , Osteogênese
10.
Nat Commun ; 15(1): 1789, 2024 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-38413608

RESUMO

Out-of-plane fluctuations, also known as stochastic displacements, of biological membranes play a crucial role in regulating many essential life processes within cells and organelles. Despite the availability of various methods for quantifying membrane dynamics, accurately quantifying complex membrane systems with rapid and tiny fluctuations, such as mitochondria, remains a challenge. In this work, we present a methodology that combines metal/graphene-induced energy transfer (MIET/GIET) with fluorescence correlation spectroscopy (FCS) to quantify out-of-plane fluctuations of membranes with simultaneous spatiotemporal resolution of approximately one nanometer and one microsecond. To validate the technique and spatiotemporal resolution, we measure bending undulations of model membranes. Furthermore, we demonstrate the versatility and applicability of MIET/GIET-FCS for studying diverse membrane systems, including the widely studied fluctuating membrane system of human red blood cells, as well as two unexplored membrane systems with tiny fluctuations, a pore-spanning membrane, and mitochondrial inner/outer membranes.


Assuntos
Grafite , Humanos , Espectrometria de Fluorescência/métodos , Membrana Celular/fisiologia , Membranas , Transferência de Energia
11.
BMC Biol ; 22(1): 46, 2024 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-38414038

RESUMO

Membranes are protein and lipid structures that surround cells and other biological compartments. We present a conceptual model wherein all membranes are organized into structural and functional zones. The assembly of zones such as receptor clusters, protein-coated pits, lamellipodia, cell junctions, and membrane fusion sites is explained to occur through a protein-lipid code. This challenges the theory that lipids sort proteins after forming stable membrane subregions independently of proteins.


Assuntos
Proteínas de Transporte , Proteolipídeos , Proteolipídeos/metabolismo , Membranas/metabolismo , Proteínas de Transporte/metabolismo , Membrana Celular/metabolismo
12.
J Vis Exp ; (203)2024 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-38314838

RESUMO

The cell membrane is crucial for cell survival, and ensuring its integrity is essential as the cell experiences injuries throughout its entire life cycle. To prevent damage to the membrane, cells have developed efficient plasma membrane repair mechanisms. These repair mechanisms can be studied by combining confocal microscopy and nanoscale thermoplasmonics to identify and investigate the role of key proteins, such as annexins, involved in surface repair in living cells and membrane model systems. The puncturing method employs a laser to induce highly localized heating upon nanoparticle irradiation. The use of near-infrared light minimizes phototoxicity in the biological sample, while the majority of the absorption takes place in the near-infrared resonant plasmonic nanoparticle. This thermoplasmonic method has been exploited for potential photothermal and biophysical research to enhance the understanding of intracellular mechanisms and cellular responses through vesicle and cell fusion studies. The approach has shown to be complementary to existing methods for membrane disruption, such as mechanically, chemically, or optically induced injuries, and provides a high level of control by inflicting extremely localized injuries. The extent of the injury is limited to the vicinity of the spherical nanoparticle, and no detrimental damage occurs along the beam path as opposed to pulsed lasers using different wavelengths. Despite certain limitations, such as the formation of nanobubbles, the thermoplasmonic method offers a unique tool for investigating cellular responses in plasma membrane repair in an almost native environment without compromising cell viability. When integrated with confocal microscopy, the puncturing method can provide a mechanistic understanding of membrane dynamics in model membrane systems as well as quantitative information on protein responses to membrane damage, including protein recruitment and their biophysical function. Overall, the application of this method to reduced model systems can enhance our understanding of the intricate plasma membrane repair machinery in living cells.


Assuntos
Nanopartículas , Membrana Celular/metabolismo , Membranas , Sobrevivência Celular , Raios Infravermelhos
13.
Langmuir ; 40(6): 2809-2814, 2024 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-38307088

RESUMO

Inspired by the structures and functions of natural channel proteins that selectively permeate ions and molecules across biological membranes, synthetic molecules capable of self-assembling into supramolecular nanotubes within the hydrophobic layer of the membranes have been designed and their material permeation properties have been studied. More recently, synthetic chemists have ventured to incorporate fluorine atoms, elements rarely found in natural proteins, into the structure of synthetic channels and discovered anomalous transmembrane material permeation properties. In this Perspective, the author provides a brief overview of recent advances in the development of fluorinated nanochannels and possible directions for the future.


Assuntos
Nanotubos , Proteínas , Nanotubos/química , Membrana Celular , Membranas
14.
Sci Rep ; 14(1): 3553, 2024 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-38347108

RESUMO

Bioactive material concepts for targeted therapy have been an important research focus in regenerative medicine for years. The aim of this study was to investigate a proof-of-concept composite structure in the form of a membrane made of natural silk fibroin (SF) and extracellular vesicles (EVs) from gingival fibroblasts. EVs have multiple abilities to act on their target cell and can thus play crucial roles in both physiology and regeneration. This study used pH neutral, degradable SF-based membranes, which have excellent cell- and tissue-specific properties, as the carrier material. The characterization of the vesicles showed a size range between 120 and 180 nm and a high expression of the usual EV markers (e.g. CD9, CD63 and CD81), measured by nanoparticle tracking analysis (NTA) and single-EV flow analysis (IFCM). An initial integration of the EVs into the membrane was analyzed using scanning and transmission electron microscopy (SEM and TEM) and vesicles were successfully detected, even if they were not homogeneously distributed in the membrane. Using direct and indirect tests, the cytocompatibility of the membranes with and without EVs could be proven and showed significant differences compared to the toxic control (p < 0.05). Additionally, proliferation of L929 cells was increased on membranes functionalized with EVs (p > 0.05).


Assuntos
Vesículas Extracelulares , Fibroínas , Nanopartículas , Fibroínas/metabolismo , Vesículas Extracelulares/metabolismo , Membranas , Nanopartículas/química , Fibroblastos
15.
Methods Mol Biol ; 2772: 353-370, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38411828

RESUMO

Confocal laser scanning microscopy (CLSM) is an advanced microscopy technique based on fluorescence technology which produces sharp images of a specimen in a single focal plane. The optical sectioning by CLSM allows to have z-stacks which can be further processed into 3D reconstructions. These then provide the option of variable perspectives and additional precise data evaluation on structural and anatomical alterations. Here, we used CLSM to image the thylakoids of cyanobacteria and the endoplasmic reticulum (ER) in moss protonemata as an example. Then, out of the confocal z-stacks, we create 3D constructions of the membranes and their alterations to present a holistic, structural view from different angles.


Assuntos
Retículo Endoplasmático , Imageamento Tridimensional , Técnicas Histológicas , Membranas , Microscopia Confocal
16.
Food Res Int ; 180: 114074, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38395577

RESUMO

Low-temperature (9-12 °C) pulsed electric field (PEF) was investigated in milk before cream separation at different intensities (9-27 kV/cm, 66 µs, 16-28 kJ/L) regarding its potential to render processing more sustainable, retain a high physico-chemical quality, enhance functional properties, and gently modify the structure of the milk fat globule membrane (MFGM). Cream volume per L milk were most efficiently increased by 31 % at the lowest PEF intensity in comparison to untreated milk and cream (P < 0.05). Untreated and PEF-treated milk and obtained cream were assessed with compositional (fat, protein, casein, lactose, and total solids content) and particle size distribution analyses, showing no significant differences (P ≥ 0.05) and, thus, indicating retention of 'native-like' product quality. Overrun and stability of cream, whipped for 20 and 60 s at 15000 rpm using a high-shear mixer, were improved most notably by the lowest and the highest PEF intensities, achieving up to 69 % enlarged overrun and up to 22 % higher stability, respectively (P < 0.05), than in untreated whipped cream. Protein component analyses for milk and cream were carried out by sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Noticeable differences between untreated and PEF-treated milk were not observed, but the SDS-PAGE results for cream showed noticeably different bands for some of the protein components, indicating structural changes in MFGM-, whey-, and phospho-proteins due to PEF and/or separator processing effects. More intense bands of xanthine oxidase, xanthine dehydrogenase, butyrophilin, bovine serum albumine, adipophilin (ADPH), and glycoproteins PAS6/7 were observed specifically at 21 kV/cm. Gentle electroporation of both MFGM layers by PEF was determined based on the changes in MFGM monolayer components, such as ADPH and PAS 6/7, exhibiting intensified bands. PEF intensity-dependent impact on the structure of MFGM and casein, leading to a reconfiguration of the cream matrix due to different structuring interactions among proteins, among milk fat globules, and between fat and protein components, was suggested. Overall, low-temperature PEF applied at different intensities showed great potential for gentle, efficient, and functional properties-tailored dairy processing and may also enable effective extraction of highly bioactive ingredients from dairy sources.


Assuntos
Caseínas , Leite , Animais , Caseínas/química , Leite/química , Proteínas do Soro do Leite/análise , Membranas , Soro do Leite
17.
Molecules ; 29(4)2024 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-38398572

RESUMO

Professor Carlos Gutiérrez-Merino, a prominent scientist working in the complex realm of biological membranes, has made significant theoretical and experimental contributions to the field. Contemporaneous with the development of the fluid-mosaic model of Singer and Nicolson, the Förster resonance energy transfer (FRET) approach has become an invaluable tool for studying molecular interactions in membranes, providing structural insights on a scale of 1-10 nm and remaining important alongside evolving perspectives on membrane structures. In the last few decades, Gutiérrez-Merino's work has covered multiple facets in the field of FRET, with his contributions producing significant advances in quantitative membrane biology. His more recent experimental work expanded the ground concepts of FRET to high-resolution cell imaging. Commencing in the late 1980s, a series of collaborations between Gutiérrez-Merino and the authors involved research visits and joint investigations focused on the nicotinic acetylcholine receptor and its relation to membrane lipids, fostering a lasting friendship.


Assuntos
Lipídeos de Membrana , Receptores Nicotínicos , Membrana Celular/metabolismo , Lipídeos de Membrana/química , Transferência Ressonante de Energia de Fluorescência , Membranas/metabolismo , Receptores Nicotínicos/metabolismo
18.
Langmuir ; 40(9): 4719-4731, 2024 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-38373285

RESUMO

Transmembrane asymmetry is ubiquitous in cells, particularly with respect to lipids, where charged lipids are mainly restricted to one monolayer. We investigate the influence of anionic lipid asymmetry on the stability of giant unilamellar vesicles (GUVs), minimal plasma membrane models. To quantify asymmetry, we apply the fluorescence quenching assay, which is often difficult to reproduce, and caution in handling the quencher is generally underestimated. We first optimize this assay and then apply it to GUVs prepared with the inverted emulsion transfer protocol by using increasing fractions of anionic lipids restricted to one leaflet. This protocol is found to produce highly asymmetric bilayers but with ∼20% interleaflet mixing. To probe the stability of asymmetric versus symmetric membranes, we expose the GUVs to porating electric pulses and monitor the fraction of destabilized vesicles. The pulses open macropores, and the GUVs either completely recover or exhibit leakage or bursting/collapse. Residual oil destabilizes porated membranes, and destabilization is even more pronounced in asymmetrically charged membranes. This is corroborated by the measured pore edge tension, which is also found to decrease with increasing charge asymmetry. Using GUVs with imposed transmembrane pH asymmetry, we confirm that poration-triggered destabilization does not depend on the approach used to generate membrane asymmetry.


Assuntos
Lipídeos , Lipossomas Unilamelares , Membrana Celular/metabolismo , Lipossomas Unilamelares/química , Membranas/metabolismo , Bicamadas Lipídicas/química
20.
PLoS One ; 19(2): e0299169, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38422081

RESUMO

Prokaryotic chromosomes contain numerous small open reading frames (ORFs) of less than 200 bases. Since high-throughput proteomics methods often miss proteins containing fewer than 60 amino acids, it is difficult to decern if they encode proteins. Recent studies have revealed that many small proteins are membrane proteins with a single membrane-anchoring α-helix. As membrane anchoring or transmembrane motifs are accurately identifiable with high confidence using computational algorithms like Phobius and TMHMM, small membrane proteins (SMPS) can be predicted with high accuracy. This study employed a systematic approach, utilizing well-verified algorithms such as Orfipy, Phobius, and Blast to identify SMPs in prokaryotic organisms. Our main search parameters targeted candidate SMPs with an open reading frame between 60-180 nucleotides, a membrane-anchoring or transmembrane region 15 and 30 amino acids long, and sequence conservation among other microorganisms. Our findings indicate that each prokaryote possesses many SMPs, with some identified in the intergenic regions of currently annotated chromosomes. More extensively studied microorganisms, such as Escherichia coli and Bacillus subtilis, have more SMPs identified in their genomes compared to less studied microorganisms, suggesting the possibility of undiscovered SMPs in less studied microorganisms. In this study, we describe the common SMPs identified across various microorganisms and explore their biological roles. We have also developed a software pipeline and an accompanying online interface for discovering SMPs (http://cs.indstate.edu/pro-smp-finder). This resource aims to assist researchers in identifying new SMPs encoded in microbial genomes of interest.


Assuntos
Antifibrinolíticos , Proteínas de Membrana , Proteínas de Membrana/genética , Membranas , Algoritmos , Aminoácidos , Escherichia coli/genética
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