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Hypothesis Atomistically-detailed models of surfactants provide quantitative information on the molecular interactions and spatial distributions at fluid interfaces. Hence, it should be possible to extract from this information, macroscopical thermophysical properties such as interfacial tension, critical micelle concentrations and the relationship between these properties and the bulk fluid surfactant concentrations. Simulations and Experiments Molecular-scale interfacial of systems containing n-dodecyl ß-glucoside (APG12) are simulated using classical molecular dynamics. The bulk phases and the corresponding interfacial regions are all explicitly detailed using an all-atom force field (PCFF+). During the simulation, the behaviour of the interface is analyzed geometrically to obtain an approximated value of the critical micelle concentration (CMC) in terms of the surfactant area number density and the interfacial tension is assessed through the analysis of the forces amongst molecules. New experimental determinations are reported for the surface tension of APG12 at the water/air and at the water/n-decane interfaces. Findings We showcase the application of a thermodynamic framework that inter-relates interfacial tensions, surface densities, CMCs and bulk surfactant concentrations, which allows the in silico quantitative prediction of interfacial tension isotherms.
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HYPOTHESIS: Amphiphilic diblock copolymers are known to increase the surfactant's efficiency to stabilize microemulsion, leading to higher structural order and monolayer rigidity. We thus seek to evaluate whether the addition of such polymers alters the shear behavior of bicontinuous microemulsions, in particular, their shear transformation towards lamellar structures. EXPERIMENTS: We examine the initial structure and shear response of bicontinuous /n-octane//PEP5-b-PEO5 microemulsions by coupling microfluidics with small-angle neutron scattering (SANS), attaining wall shear rates in excess of . The azimuthal analysis of the obtained 2D scattering patterns allows us to follow their structural transformation by means of the degree of anisotropy. FINDINGS: The amphiphilic diblock copolymer promotes the shear-induced transformation of bicontinuous microemulsions, resulting in up to â¼ higher degrees of anisotropy than for corresponding polymer-free microemulsions. The increased shear response observed with increasing polymer content is rationalized by combining the influence of domain size and viscosity with the stability limits of the bicontinuous microemulsion in the isothermal phase diagram. As a result, a consistent description of the degree of anisotropy is obtained, enabling the prediction of the shear-induced bicontinuous-to-lamellar transformation.
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Ultrathin electrospun poly (l-lactide-co-dl-lactide) nanofibrous membranes coated with fibronectin were explored as scaffolds for the ex vivo cultivation of limbal epithelial cells (LECs) for the treatment of limbal stem cell deficiency. The developed scaffolds were compared with the "gold-standard" fibrin gel. The resulting membranes composed of nanofibers possessed a very low thickness of 4 µm and allowed very good optical transparency in the wet state. The fibronectin-coated nanofibrous scaffolds demonstrated LEC expansion and successful cultivation similar to that on fibrin gel. Unlike the regular cobblestone epithelial cell morphology on the fibrin gel, the nanofibrous scaffold presented a mostly irregular epithelial morphology with a shift to a mesenchymal phenotype, as confirmed by the upregulation of profibroblastic genes: ACTA2 (p = 0.023), FBLN1 (p < 0.001), and THY1 (p < 0.001). Both culture conditions revealed comparable expression of stem cell markers, including KLF4, ΔNp63α and ABCG2, emphasizing the promise of polylactide-based nanofibrous membranes for further investigations.
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HYPOTHESIS: The physical and mechanical properties of triacylglycerols (TAGs), or 'fats', depend on their composition and thermal history which, in turn, impact crystal structure and morphology. We examine whether thermal history can be mechanistically related to film removal by a surfactant solution. EXPERIMENTS: Model TAG mixtures, comprising triolein:tripalmitin:tristearin 0.5:0.3:0.2, were subjected to a range of cooling profiles from the melt (0.5-80°C/min, Newtonian and annealed), and the resulting solid films characterised by microscopy, X-ray diffraction, infrared spectroscopy, and contact angle measurements. Film removal from a model glass substrate by an aqueous surfactant solution of sodium dodecylsulphate and dodecyldimethylamine oxide at room temperature fixed at 25°C was examined under quiescent flow conditions. FINDINGS: Quantitative relations are established between TAG cooling profile, crystal structure and morphology, surface energy γSFE, and removal (or 'cleaning'). In general, films cooled slowly from the melt yield heterogeneous morphologies with predominantly ß1' phase, higher polar γSFE, and faster removal timescales. By contrast, rapid cooling results in homogeneous films, rich in ß2' phase, low polar γSFE, and long removal times. Our results elucidate the non-trivial impact of TAG thermal history, connecting the multiscale semi-crystalline structure to surface energy, and eventually to film delamination by micellar solutions.
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In bacteria, intracellular K+ is involved in the regulation of membrane potential, cytosolic pH, and cell turgor as well as in spore germination, environmental adaptation, cell-to-cell communication in biofilms, antibiotic sensitivity, and infectivity. The second messenger cyclic-di-AMP (c-di-AMP) has a central role in modulating the intracellular K+ concentration in many bacterial species, controlling transcription and function of K+ channels and transporters. However, our understanding of how this regulatory network responds to c-di-AMP remains poor. We used the RCK (Regulator of Conductance of K+) proteins that control the activity of Ktr channels in Bacillus subtilis as a model system to analyze the regulatory function of c-di-AMP with a combination of in vivo and in vitro functional and structural characterization. We determined that the two RCK proteins (KtrA and KtrC) are neither physiologically redundant or functionally equivalent. KtrC is the physiologically dominant RCK protein in the regulation of Ktr channel activity. In explaining this hierarchical organization, we found that, unlike KtrA, KtrC is very sensitive to c-di-AMP inactivation and lack of c-di-AMP regulation results in RCK protein toxicity, most likely due to unregulated K+ flux. We also found that KtrC can assemble with KtrA, conferring c-di-AMP regulation to the functional KtrA/KtrC heteromers and potentially compensating KtrA toxicity. Altogether, we propose that the central role of c-di-AMP in the control of the K+ machinery, by modulating protein levels through gene transcription and by regulating protein activity, has determined the evolutionary selection of KtrC as the dominant RCK protein, shaping the hierarchical organization of regulatory components of the K+ machinery.
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Bacillus subtilis , Proteínas Bacterianas , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Bacillus subtilis/metabolismo , Bacillus subtilis/genética , Potasio/metabolismo , Regulación Bacteriana de la Expresión Génica , Fosfatos de Dinucleósidos/metabolismo , Canales de Potasio/metabolismo , Canales de Potasio/genéticaRESUMEN
We investigate the structure and interactions of a model anionic/amphoteric mixed surfactant micellar system, namely, sodium dodecyl sulfate (SDS) and N,N-dimethyldodecylamine N-oxide (DDAO), employing SANS, FTIR, DLS, and pH measurements, in the range 0.1-100 mM total surfactant concentration and 0-100% DDAO. Increasing surfactant concentration is found to elongate the prolate ellipsoid micelles (RPolar â¼ 25-40 Å), accompanied by up to a 6-fold increase in micellar charge. The surfactant synergy, in terms of micellar charge and size, diffusion coefficient, solution pH, and headgroup interactions, was found to vary with concentration. At lower concentrations (≤50 mM), the SDS-DDAO ratio of maximum synergy is found to be asymmetric (at 65-85% DDAO), which is rationalized using regular solution theory, suggesting an equilibrium between Na+ dissociation, DDAO protonation, and counterion concentration. At higher concentrations, maximum synergy shifts toward the equimolar ratio. Overall, our study expands and unifies previous reports, providing a comprehensive understanding for this model, synergetic mixed micellar system.
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Slow deactivation is a critical property of voltage-gated K+ channels encoded by the human Ether-à-go-go-Related Gene 1 (hERG). hERG1 channel deactivation is modulated by interactions between intracellular N-terminal Per-Arnt-Sim (PAS) and C-terminal cyclic nucleotide-binding homology (CNBh) domains. The PAS domain is multipartite, comprising a globular domain (gPAS; residues 26-135) and an N-terminal PAS-cap that is further subdivided into an initial unstructured "tip" (residues 1-12) and an amphipathic α-helical region (residues 13-25). Although the PAS-cap tip has long been considered the effector of slow deactivation, how its position near the gating machinery is controlled has not been elucidated. Here, we show that a triad of hydrophobic interactions among the gPAS, PAS-cap α helix, and the CNBh domains is required to support slow deactivation in hERG1. The primary sequence of this "hydrophobic nexus" is highly conserved among mammalian ERG channels but shows key differences to fast-deactivating Ether-à-go-go 1 (EAG1) channels. Combining sequence analysis, structure-directed mutagenesis, electrophysiology, and molecular dynamics simulations, we demonstrate that polar serine substitutions uncover an intermediate deactivation mode that is also mimicked by deletion of the PAS-cap α helix. Molecular dynamics simulation analyses of the serine-substituted channels show an increase in distance among the residues of the hydrophobic nexus, a rotation of the intracellular gating ring, and a retraction of the PAS-cap tip from its receptor site near the voltage sensor domain and channel gate. These findings provide compelling evidence that the hydrophobic nexus coordinates the respective components of the intracellular gating ring and positions the PAS-cap tip to control hERG1 deactivation gating.
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Interacciones Hidrofóbicas e Hidrofílicas , Animales , Humanos , Secuencia de Aminoácidos , Canal de Potasio ERG1/metabolismo , Canal de Potasio ERG1/química , Canal de Potasio ERG1/genética , Canales de Potasio Éter-A-Go-Go/química , Canales de Potasio Éter-A-Go-Go/metabolismo , Canales de Potasio Éter-A-Go-Go/genética , Células HEK293 , Espacio Intracelular/metabolismo , Activación del Canal Iónico , Dominios ProteicosRESUMEN
The aim of this paper is to review the topic of neonatal fractures of the upper limb, describing the different types of fractures focusing on the etiology, epidemiology, risk factors, clinical approach, diagnosis, treatment and prognosis of these injuries. We included all types of research studies, both experimental and observational, published in English, French, Portuguese and Spanish. The information was obtained using the keywords neonatal upper limb fracture, clavicle fracture or humerus fracture from the following resources: MEDLINE database, Embase® database and LILACS database. Other resources such as hand searches of the references of retrieved literature and authoritative texts, personal and hospital libraries searching for texts on upper limb neonatal fractures, discussions with experts in the field of upper limb neonatal fractures and personal experience, were also considered for the completion of the article.Neonatal fractures of the upper limb are consensually considered to have a good prognosis and no long-term sequelae. Conservative treatment is the option in the vast majority of the fractures and is associated with excellent results, with good healing, full range of motion, adequate remodeling without obvious deformity, neurologic impairment or functional implications.
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Fracturas Óseas , Fracturas del Húmero , Recién Nacido , Humanos , Fracturas Óseas/diagnóstico , Fracturas Óseas/etiología , Fracturas Óseas/terapia , Extremidad Superior , Clavícula , Pronóstico , Factores de RiesgoRESUMEN
A case series of the use of amniotic membrane (AM) for treating chronic nonhealing wounds. It presents five cases of polymorbid patients with a total of nine chronic nonhealing wounds. The patient group consisted of four men and one woman with various comorbidities, aged 45-72 years. The mean initial wound size was 15.8 cm2, and the mean time from the onset of the wound to the first application of AM was 122 weeks. The wounds were caused by chronic venous insufficiency and/or peripheral arterial disease. Wounds were treated in a standardized protocol. AM was applied weekly in the first month and then every two weeks. Photo documentation of the wound and microbiological colonization was carried out at each visit. In three out of five patients, the AM treatment effectively promoted healing up to complete wound closure. In two cases, the wounds stayed unhealed despite numerous AM applications. Pain relief was noted in all patients. The success of the treatment was closely tied to patient factors, such as adherence to the prescribed treatment regimen and individual patient characteristics. In some cases, treatment failure was observed, possibly due to underlying comorbidities, wound parameters, or poor patient compliance. AM treatment has the potential to become a viable treatment option for these nonhealing wounds. However, the effectiveness of the treatment may be influenced by various patient factors and the underlying cause of the wound. Therefore, it is crucial to have an individualized treatment plan that considers these particular factors.
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Amnios , Cicatrización de Heridas , Masculino , Femenino , Humanos , Resultado del Tratamiento , Criopreservación/métodos , Estudios RetrospectivosRESUMEN
We investigate the drying of isolated polymer solution droplets, employing acoustic levitation, and demonstrate the spontaneous generation of breath figures (BF) on the resulting polymer particles and capsules (â¼5-1000 µm) with controlled surface pore arrays (<1-20 µm). By contrast with supported polymer thin films, the evaporative cooling experienced by suspended droplets suffices to yield ubiquitous BF formation, owing to their thermal insulation and the synchronous condensation and self-assembly of water microdroplets, accompanied by capsule skin formation and kinetic arrest. A simple model describes simultaneously the radius and temperature evolution along the droplet-to-particle transformation, and the scaling of surface pore dimensions, with environmental parameters. The generality of the approach is demonstrated with a range of model polymers, and the coupled roles of solution thermodynamics and droplet environment are shown to permit the facile design of capsules with tunable transport and dissolution kinetics.
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We investigate the solution structures of model sodium dodecyl sulfate/octanol/brine ternary mixtures across the lamellar (Lα), vesicle (L4) and micellar (L1) phases employing small angle neutron scattering (SANS), optical microscopy and nuclear magnetic resonance (NMR). Specifically, we examine the effect of co-surfactant octanol (0.2-9.48 w/v%) and temperature (25-65 °C) along dilution lines at fixed octanol : SDS ratios (0.08-1.21). A transition from Lα to sponge phase (L3) above 35 °C is found along the octanol : SDS = 1.21 isopleth, with phase coexistence above Ï ≈ 0.14 weight fraction of surfactant and co-surfactant. The lamellar bilayers swell upon dilution, with an approximately linear increase of d-spacing, accompanied by a decrease of the Caillé parameter, indicative of greater membrane rigidity. At a lower octanol : SDS ratio of 0.62, coexistence of oblate micelles and vesicles is observed with preferential formation of vesicles at low concentrations. Dilution of the L1 phase, along octanol : SDS = 0.08, results in elongated micelles, as the NaCl : SDS ratio increases, while higher temperatures favour the formation of less elongated micelles. Our results provide a detailed map of the equilibrium structures found in the Lα vicinity of this extensively investigated flow-responsive surfactant system.
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PURPOSE: The Hugo™ RAS system is a novel robotic platform with innovative features. However, there are currently no available data on extraperitoneal robot-assisted radical prostatectomy (RARP) performed using this system. The objective of this study is to describe the surgical setup and assess the safety and feasibility of the extraperitoneal approach in robotic radical prostatectomy with the Hugo™ RAS system. METHODS: Sixteen consecutive patients diagnosed with localized prostate cancer underwent extraperitoneal RARP ± lymph node dissection at our institution, between March and May 2023. All RARP procedures were performed extraperitoneal with a modular four-arm configuration. The focus was to describe the operative room setup, trocar placement, tilt and docking angles and evaluate the safety and feasibility of this approach with this robotic platform. Secondary outcomes recorded included, total operative time, console time, estimated bleeding, intra- and postoperative complications, and length of stay after surgery. A descriptive analysis was conducted. RESULTS: We report on the first sixteen cases of extraperitoneal robot-assisted radical prostatectomy performed with the new Hugo™ RAS system. All procedures were completed, without the need for conversion or placement of additional ports. No intraoperative complications or major technical failures that would prevent the completion of surgery were recorded. The median operative time was 211 min (IQR 180-277), and the median console time was 152 min (IQR 119-196). The mean docking time was 4.6 min (IQR 4.1-5.2). The median estimated blood loss and the median time to remove the vesical catheter were 200 mL (IQR 150-400) and 8 days (IQR 7-8), respectively. The median length of stay was 2 days (IQR 2-2). Only one minor complication was registered in the first 30 days. CONCLUSION: This study provides evidence of the safety and feasibility of the extraperitoneal approach in RARP with the Hugo™ RAS system. The description of the surgical setup in terms of trocar placement, arm-cart disposition, tilt and docking angles offers valuable information for surgeons interested in adopting this surgical approach with the Hugo™ RAS platform.
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Laparoscopía , Neoplasias de la Próstata , Procedimientos Quirúrgicos Robotizados , Robótica , Masculino , Humanos , Procedimientos Quirúrgicos Robotizados/métodos , Próstata/cirugía , Prostatectomía/métodos , Neoplasias de la Próstata/cirugía , Laparoscopía/métodosRESUMEN
BACKGROUND: Biochemical markers and imaging tests have been used with the aim of stratifying the risk and detecting atrial fibrosis. Speckle-tracking echocardiography (STE) is used for the detection of atrial fibrosis and Gal-3 provides an important prognostic value. The objective of the study was to assess the association between atrial fibrosis markers and serum levels, genetic polymorphisms and genic expression of galectin-3. METHODS: 206 patients with permanent AF and 70 patients with paroxysmal AF were included in the study. Real time PCR (TaqMan) system was used to study SNPs rs4652 and 4644 of the gene LGALS3. Serum levels of Gal-3 were determined by ELISA and STE was performed to assess fibrosis. RESULTS: Mean age of individuals with permanent AF was 66.56 ± 12 years. As for the echocardiography results, those patients showed an decrease in the following parameters peak atrial longitudinal strain (PALS) (p = 0.002) when compared to the same parameters from the paroxysmal AF group of patients. There was a correlation between serum levels of Gal-3 and PALS in the group of patients with permanent AF; the lower the levels of gal-3, the lower the LA strain (r = 0.24; p = 0.01). CONCLUSIONS: Echocardiographic findings showed association with the groups, and with serum levels of Gal-3 in patients with permanent AF. The distribution of allelic and genotypic frequencies, and of the haplotypes of polymorphism LGALS3 rs4652 and rs4644 did not present statistical variation, which suggests that those SNPs are not associated with the AF clinical forms (permanent and paroxysmal).
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Fibrilación Atrial , Humanos , Persona de Mediana Edad , Anciano , Fibrilación Atrial/diagnóstico por imagen , Fibrilación Atrial/genética , Galectina 3/genética , Atrios Cardíacos/diagnóstico por imagen , Atrios Cardíacos/patología , Ecocardiografía/métodos , FibrosisRESUMEN
â¢Giant and solitary polyps evolve with anemia. â¢EUS is an important tool for stage and manage this disease. â¢Endoscopic treatment is the best treatment choice. â¢Supplementary video available on this case report.
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Anemia , Hamartoma , Síndrome de Peutz-Jeghers , Pólipos , Humanos , Síndrome de Peutz-Jeghers/complicaciones , Síndrome de Peutz-Jeghers/cirugía , Pólipos/complicaciones , Pólipos/cirugía , Endoscopía , Hamartoma/complicaciones , Hamartoma/cirugía , Anemia/etiologíaRESUMEN
We investigate the thermodynamics of a highly interacting blend of poly(cyclohexyl methacrylate)/deuterated poly(styrene) (PCHMA/dPS) with small-angle neutron scattering (SANS). This system is experimentally challenging due to the proximity of the blend phase boundary (>200 °C) and degradation temperatures. To achieve the large wavenumber q-range and flux required for kinetic experiments, we employ a SANS diffractometer in time-of-flight (TOF) mode at a reactor source and ancillary microscopy, calorimetry, and thermal gravimetric analysis. Isothermal SANS data are well described by random-phase approximation (RPA), yielding the second derivative of the free energy of mixing (Gâ³), the effective interaction (χÌ ) parameter, and extrapolated spinodal temperatures. Instead of the Cahn-Hilliard-Cook (CHC) framework, temperature (T)-jump experiments within the one-phase region are found to be well described by the RPA at all temperatures away from the glass transition temperature, providing effectively near-equilibrium results. We employ CHC theory to estimate the blend mobility and Gâ³(T) conditions where such an approximation holds. TOF-SANS is then used to precisely resolve Gâ³(T) and χÌ (T) during T-jumps in intervals of a few seconds and overall timescales of a few minutes. PCHMA/dPS emerges as a highly interacting partially miscible blend, with a steep dependence of Gâ³(T) [mol/cm3] = -0.00228 + 1.1821/T [K], which we benchmark against previously reported highly interacting lower critical solution temperature (LCST) polymer blends.
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Coupling microfluidics and small-angle neutron scattering (SANS), we investigate the influence of shear flow on a model bicontinuous microemulsion of D2O/n-octane/C10E4, examining the role of membrane volume fraction in the transformation towards a lamellar structure. We employ a contraction-expansion geometry with flow velocities in excess of 10 m s-1 and spatially map the microfluidic field using a small SANS beam, illuminating down to 10 nL sample volumes. The shear-induced, progressive, bicontinuous-to-lamellar transition is found to be promoted by additional extensional flow (>103 s-1), while fast relaxation kinetics (<2 ms) return the scattering pattern to isotropic shortly after the constriction. Further, increasing the domain size of the bicontinuous structure (determined by the membrane volume fraction) appears to amplify its response to shear. Hence, the structural changes within the dilute bicontinuous microemulsions simply scale with the volume fraction of the membrane. By contrast, the stronger response of the microemulsion with the smallest domain size, located near the bicontinuous/lamellar coexistence, indicates an influence of an already more ordered structure with fewer passages. Our findings provide insight into the high shear behaviour of microemulsions of both academic and industrial relevance.
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Septins are cytoskeletal proteins interacting with the inner plasma membrane and other cytoskeletal partners. Being key in membrane remodeling processes, they often localize at specific micrometric curvatures. To analyze the behavior of human septins at the membrane and decouple their role from other partners, we used a combination of bottom-up in vitro methods. We assayed their ultrastructural organization, their curvature sensitivity, as well as their role in membrane reshaping. On membranes, human septins organize into a two-layered mesh of orthogonal filaments, instead of generating parallel sheets of filaments observed for budding yeast septins. This peculiar mesh organization is sensitive to micrometric curvature and drives membrane reshaping as well. The observed membrane deformations together with the filamentous organization are recapitulated in a coarse-grained computed simulation to understand their mechanisms. Our results highlight the specific organization and behavior of animal septins at the membrane as opposed to those of fungal proteins.
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Citoesqueleto , Septinas , Animales , Humanos , Septinas/genética , Membranas , Membrana Celular , BioensayoRESUMEN
We report the coupling of dynamic light scattering (DLS) in microfluidics, using a contact-free fibre-optic system, enabling the under-flow characterisation of a range of solutions, dispersions, and structured fluids. The system is evaluated and validated with model systems, specifically micellar and (dilute) polymer solutions, and colloidal dispersions of different radii (â¼1-100 nm). A systematic method of flow-DLS analysis is examined as a function of flow velocity (0-16 cm s-1), and considerations of the relative contribution of 'transit' and 'Brownian' terms enable the identification of regions where (i) a quiescent approximation suffices, (ii) the flow-DLS framework holds, as well as (iii) where deviations are found, until eventually (iv) the convection dominates. We investigate practically relevant, robust setups, namely that of a capillary connected to microdevice, as well as direct measurement on a glass microdevice, examining the role of capillary dimensions and challenges of optical alignment. We conclude with a demonstration of a continuous flow measurement of a binary surfactant/salt solution, whose micellar dimensions vary with composition, characterised with hundreds of data points (every â¼5 s) and adequate statistics, within a few minutes.
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This study aimed to evaluate the efficacy of cryopreserved amniotic membrane (AM) grafts in chronic wound healing, including the mean percentage of wound closure per one AM application, and to determine whether the healing efficiency differs between AM grafts obtained from different placentas. A retrospective study analyzing inter-placental differences in healing capacity and mean wound closure after the application of 96 AM grafts prepared from nine placentas. Only the placentas from which the AM grafts were applied to patients suffering from long-lasting non-healing wounds successfully healed by AM treatment were included. The data from the rapidly progressing wound-closure phase (p-phase) were analyzed. The mean efficiency for each placenta, expressed as an average of wound area reduction (%) seven days after the AM application (baseline, 100%), was calculated from at least 10 applications. No statistical difference between the nine placentas' efficiency was found in the progressive phase of wound healing. The 7-day average wound reduction in particular placentas varied from 5.70 to 20.99% (median from 1.07 to 17.75) of the baseline. The mean percentage of wound surface reduction of all analyzed defects one week after the application of cryopreserved AM graft was 12.17 ± 20.12% (average ± SD). No significant difference in healing capacity was observed between the nine placentas. The data suggest that if there are intra- and inter-placental differences in AM sheets' healing efficacy, they are overridden by the actual health status of the subject or even the status of its individual wounds.