RESUMO
The two major subtypes of human T cells, CD4+ and CD8+, play important roles in adaptive immune response by their diverse functions. To understand the structure-function relation at the single cell level, we isolated 2483 CD4+ and 2450 CD8+ T cells from fresh human splenocytes by immunofluorescent sorting and investigated their morphologic relations to the surface CD markers by acquisition and analysis of cross-polarized diffraction image (p-DI) pairs. A deep neural network of DINet-R has been built to extract 2560 features across multiple pixel scales of a p-DI pair per imaged cell. We have developed a novel algorithm to form a matrix of Pearson correlation coefficients by these features for selection of a support cell set with strong morphologic correlation in each subtype. The p-DI pairs of support cells exhibit significant pattern differences between the two subtypes defined by CD markers. To explore the relation between p-DI features and CD markers, we divided each subtype into two groups of A and B using the two support cell sets. The A groups comprise 90.2% of the imaged T cells and classification of them by DINet-R yields an accuracy of 97.3 ± 0.40% between the two subtypes. Analysis of depolarization ratios further reveals the significant differences in molecular polarizability between the two subtypes. These results prove the existence of a strong structure-function relation for the two major T cell subtypes and demonstrate the potential of diffraction imaging flow cytometry for accurate and label-free classification of T cell subtypes.
Assuntos
Aprendizado Profundo , Linfócitos T CD4-Positivos , Linfócitos T CD8-Positivos , Citometria de Fluxo/métodos , Humanos , Redes Neurais de ComputaçãoRESUMO
Poly(vinyl alcohol) (PVA) is a cytocompatible synthetic polymer and has been commonly used to prepare hydrogels. Bile acids and ß-cyclodextrin are both natural compounds and they form stable host-guest inclusion complexes. They are attached covalently onto a low molecular weight PVA separately. Self-healing hydrogels can be easily formed by mixing the aqueous solutions of these PVA based polymers. The mechanical properties of the hydrogels can be tuned by varying the molar fractions of bile acid units on PVA. The dynamic inclusion complexation of the host-guest pair of the hydrogel allows the self-healing rapidly under ambient atmosphere and their mechanical properties could recover their original values in 1 min after incision. These PVA based polymers exhibited the good cytocompatibility and high hemocompatibility as shown by their biological evaluations. The use of natural compounds for host-guest interaction make such gels especially convenient to use as biomaterials, an advantage over conventional hydrogels prepared through freeze-thaw method.
Assuntos
Fibroblastos/metabolismo , Hidrogel de Polietilenoglicol-Dimetacrilato , Teste de Materiais , Álcool de Polivinil , beta-Ciclodextrinas , Animais , Fibroblastos/citologia , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacologia , Camundongos , Álcool de Polivinil/química , Álcool de Polivinil/farmacologia , beta-Ciclodextrinas/química , beta-Ciclodextrinas/farmacologiaRESUMO
Significance: Multiparameter spectrophotometry (MPS) provides a powerful tool for accurate characterization of turbid materials in applications such as analysis of material compositions, assay of biological tissues for clinical diagnosis and food safety monitoring. Aim: This work is aimed at development and validation of a rapid inverse solver based on a particle swarm optimization (PSO) algorithm to retrieve the radiative transfer (RT) parameters of absorption coefficient, scattering coefficient and anisotropy factor of a turbid sample. Approach: Monte Carlo (MC) simulations were performed to obtain calculated signals for comparison to the measured ones of diffuse reflectance, diffuse transmittance and forward transmittance. An objective function has been derived and combined with the PSO algorithm to iterate MC simulations for MPS. Results: We have shown that the objective function can significantly reduce the variance in calculated signals by local averaging of an inverse squared error sum function between measured and calculated signals in RT parameter space. For validation of the new objective function for PSO based inverse solver, the RT parameters of 20% Intralipid solutions have been determined from 520 to 1000 nm which took about 2.7 minutes on average to complete signal measurement and inverse calculation per wavelength. Conclusion: The rapid solver enables MPS to be translated into easy-to-use and cost-effective instruments without integrating sphere for material characterization by separating and revealing compositional profiles at the molecular and particulate scales.
Assuntos
Espalhamento de Radiação , Espectrofotometria , Método de Monte CarloRESUMO
Significance: As a noncontact method, imaging photoplethysmography (iPPG) may provide a powerful tool to measure pulsatile pressure wave (PPW) in superficial arteries and extract biomarkers for monitoring of artery wall stiffness. Aim: We intend to develop a approach for extraction of the very weak cardiac component from iPPG data by identifying locations of strong PPW signals with optimized illumination wavelength and determining pulse wave velocity (PWV). Approach: Monochromatic in vivo iPPG datasets have been acquired from left hands to investigate various algorithms for retrieval of PPW signals, distribution maps and waveforms, and their dependence on arterial location and wavelength. Results: A robust algorithm of pixelated independent component analysis (pICA) has been developed and combined with spatiotemporal filtering to retrieve PPW signals. Spatial distributions of PPW signals have been mapped in 10 wavelength bands from 445 to 940 nm and waveforms were analyzed at multiple locations near the palmar artery tree. At the wavelength of 850 nm selected for timing analysis, we determined PWV values from 12 healthy volunteers in a range of 0.5 to 5.8 m/s across the hand region from wrist to midpalm and fingertip. Conclusions: These results demonstrate the potentials of the iPPG method based on pICA algorithm for translation into a monitoring tool to characterize wall stiffness of superficial artery by rapid and noncontact measurement of PWV and other biomarkers within 10 s.
Assuntos
Pica , Análise de Onda de Pulso , Humanos , Análise de Onda de Pulso/métodos , Fluxo Pulsátil , Artérias/diagnóstico por imagem , Fotopletismografia , Velocidade do Fluxo SanguíneoRESUMO
Development of label-free methods for accurate classification of cells with high throughput can yield powerful tools for biological research and clinical applications. We have developed a deep neural network of DINet for extracting features from cross-polarized diffraction image (p-DI) pairs on multiple pixel scales to accurately classify cells in five types. A total of 6185 cells were measured by a polarization diffraction imaging flow cytometry (p-DIFC) method followed by cell classification with DINet on p-DI data. The averaged value and SD of classification accuracy were found to be 98.9% ± 1.00% on test data sets for 5-fold training and test. The invariance of DINet to image translation, rotation, and blurring has been verified with an expanded p-DI data set. To study feature-based classification by DINet, two sets of correctly and incorrectly classified cells were selected and compared for each of two prostate cell types. It has been found that the signature features of large dissimilarities between p-DI data of correctly and incorrectly classified cell sets increase markedly from convolutional layers 1 and 2 to layers 3 and 4. These results clearly demonstrate the importance of high-order correlations extracted at the deep layers for accurate cell classification.
Assuntos
Aprendizado Profundo , Citometria de Fluxo , Humanos , Masculino , Redes Neurais de Computação , PróstataRESUMO
Self-healing is one of the most fundamental properties of living tissues that allows them to withstand repeated damage. Self-healing hydrogels, which can spontaneously recover themselves after being ruptured, result in enhanced lifetimes for materials and open up a fascinating direction in materials science. Host-guest interactions have been widely used in the construction of self-healing hydrogels. We have emphasized the preparation and biomedical applications of self-healing hydrogels assembled by host-guest interactions, focusing on hosts of cyclodextrins and cucurbit[n]urils with various guests and tailored structures. The applications of self-healing hydrogels in biomedical fields such as drug delivery, encapsulation of cells and tissue engineering and 3D printing as well as interfacial adhesion were also summarized. At the end of this review, we propose the current challenges and future perspectives in this developing area.
Assuntos
Tecnologia Biomédica/métodos , Sistemas de Liberação de Medicamentos/métodos , Hidrogéis/química , HumanosRESUMO
The use of natural compounds to construct biomaterials, including delivery system, is an attractive strategy. In the present study, through threading functional α-cyclodextrins onto the conjugated macromolecules of poly(ethylene glycol) (PEG) and natural compound bile acid, glycopolymers of polyrotaxanes with the active targeting ability are obtained. These glycopolymers self-assemble into micelles as evidenced by dynamic light scattering and transmission electron microscopy, in which glucosamine, as an example of targeting groups, is introduced. These micelles after loading doxorubicin (DOX) exhibit the selective recognition with cancer cells 4T1. Meanwhile, the maximal half inhibitory concentration is determined to be ≈2.5 mg L-1 for the DOX-loaded micelles, close to the value of free DOX·HCl (1.9 mg L-1 ). The cumulative release of DOX at pH 5.5 is faster than at pH 7.4, which may be used as the controlled release system. This drug delivery system assembled by glycopolymers features high drug loading of DOX, superior biocompatibility. The strategy not only utilizes the micellization induced by bile acids, but also overcomes the major limitation of PEG such as the lack of targeting groups. In particular, this drug delivery platform can extend to grafting the other targeting groups, rendering this system more versatile.
Assuntos
Ácidos e Sais Biliares , Doxorrubicina , Portadores de Fármacos , Micelas , Neoplasias Experimentais/tratamento farmacológico , Rotaxanos , Animais , Ácidos e Sais Biliares/química , Ácidos e Sais Biliares/farmacocinética , Ácidos e Sais Biliares/farmacologia , Linhagem Celular Tumoral , Doxorrubicina/química , Doxorrubicina/farmacocinética , Doxorrubicina/farmacologia , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacologia , Camundongos , Células NIH 3T3 , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Coelhos , Rotaxanos/química , Rotaxanos/farmacocinética , Rotaxanos/farmacologiaRESUMO
Corneal transplantation is the widely accepted treatment to restore sight for corneal blindness. To date, because of the global donor cornea shortage, there is a need for alternatives to human donor corneas. Biocompatible collagen is an excellent candidate material for corneal repair in the view of biomimetics. Herein a class of polyrotaxane multiple aldehyde (PRA) cross-linkers based on the host-guest supramolecules of α-cyclodextrins and poly(ethylene glycol) is prepared to cross-link with collagen to fabricate materials for corneal repair. Aldehyde groups from rotaxanes and α-cyclodextrin units can synergistically improve the mechanical and optical properties of PRA cross-linked collagen membranes (Col-PRAs). Compared with counterparts cross-linked by traditional a cross-linker of 1-ethyl-3-(3-(dimethylamino)propyl) carbodiimide and N-hydroxy-succinimide, Col-PRAs have better mechanical properties, especially suture resistance as well as optical properties. In vivo lamellar keratoplasty results indicate that Col-PRAs not only can bear tight suturing on a rabbit cornea but also are prone to the remodeling of the epithelium and stroma of the cornea due to the outstanding cell adhesion and proliferation. These novel Col-PRAs exhibit great potential for use in the corneal regeneration.
RESUMO
A new and noncontact approach of multispectral reflectance imaging has been developed to inversely determine the absorption coefficient of µ a , the scattering coefficient of µs and the anisotropy factor g of a turbid target from one measured reflectance image. The incident beam was profiled with a diffuse reflectance standard for deriving both measured and calculated reflectance images. A GPU implemented Monte Carlo code was developed to determine the parameters with a conjugate gradient descent algorithm and the existence of unique solutions was shown. We noninvasively determined embedded region thickness in heterogeneous targets and estimated in vivo optical parameters of nevi from 4 patients between 500 and 950nm for melanoma diagnosis to demonstrate the potentials of quantitative reflectance imaging.
RESUMO
Two new compounds, (2R,3R,4R)-3',5'-dimethoxy-3,4,7,4'-tetrahydroxy-flavan (1) and 2-(4-hydroxy-3-methoxybenzoyl)-4-methoxy-benzaldehyde (2), together with 35 known phenolic compounds were obtained from the fruits of Amomum tsao-ko. Structures of the new compounds were elucidated on the basis of spectroscopic means, including 2D NMR, a n d high-resolution M S analysis. The isolated compounds were tested in vitro for t heircomplement-inhibitory properties against the classical pathway (CP) and alternative pathway (AP). The results showed that 14 compounds exhibited anti-complementary activities against the CP and AP with CH50 values of 0.42 - 4.43 mM and AP50 values of 0.53 -1.51 mM. Preliminary mechanism studies showed that 1,7-bis(4-hydroxyphenyl)-4(E)-hepten-3-one (8) blocked C1q, C2, C3, C4, C5 and C9 components of the complement system, and hydroquinone (15) acted on C1q, C2, C3, C5 and C9 components.
Assuntos
Amomum/química , Benzaldeídos/isolamento & purificação , Inativadores do Complemento/isolamento & purificação , Flavonoides/isolamento & purificação , Animais , Benzaldeídos/química , Medicamentos de Ervas Chinesas/química , Flavonoides/química , Frutas/química , Humanos , Masculino , Estrutura Molecular , Plantas Medicinais/química , Coelhos , OvinosRESUMO
OBJECTIVE: To better understand the epidemiology of rabies during the past ten years in Yancheng city, Jiangsu province. METHODS: Data was collected and analyzed on rabies cases in Yancheng. Density and vaccination rate on Canine, Rate of injured people bit by dogs, and the information of post-exposure prophylaxis were studied. Rabies virus in the dog brains, collected around the epidemic areas of Yancheng, were detected and analyzed. RESULTS: A total of 135 human rabies cases occurred from 1999 through 2008, and formed the second epidemic peak since 1958. Of these victims, 84% (114) were farmers. In general, the rate of people having dogs were 3% - 6% per 100 people, and the injured person-times of 100 dogs were 6.37 per year. Notably, the vaccination rate of dogs was only 20%. Of those people injured by dogs and other animals, 77% had received post-exposure treatment, and only 5% - 10% had been administered anti-rabies serum. Rabies virus antigen was found in 4 (3.6%) of 111 brain specimens among dogs collected from epidemic areas. Genetic analysis of N and G genes, which were amplified from brain specimens, indicated that these viruses belong to genotype I rabies and expressing a close relationship with the Chinese vaccine strain CTN. CONCLUSION: The large number of dogs with low vaccination rate among them, together with the incorrect and low post-exposure treatment in rural areas seemed to be responsible for the outbreak of rabies in Yancheng city.