A microparticle blood analogue suspension matching blood rheology.

The handling of blood in vitro is demanding because of ethical, economical and safety issues. Although several Newtonian and non-Newtonian blood analogues are found in the literature, few studies have used particles to mimic red blood cells (RBCs) and built an analogue with similar rheological properties of blood. This work reports the development of a blood analogue suspension composed of polydimethylsiloxane (PDMS) microparticles with an average diameter of ∼7 µm. High throughput production of PDMS particles is possible using a multi-stage membrane emulsification process; up to ∼6 mL of microparticles are manufactured in 3 hours. PDMS particles at a concentration of around 21% (w/w) at 20 °C present steady, oscillatory and extensional rheologies very similar to those of blood under physiological conditions (37 °C and ∼41% hematocrit), making them a good candidate whole blood analogue. Also, flow studies were performed in microchannels with contraction to study the cell-free layer (CFL) formation and particle deformation, achieving good qualitative results. Using the procedure developed, it is possible to obtain blood analogue fluids with a shelf life of at least 6 months.

Cell adhesion in microchannel multiple constrictions - Evidence of mass transport limitations.

Biofilm growth (fouling) in microdevices is a critical concern in several industrial, engineering and health applications, particularly in novel high-performance microdevices often designed with complex geometries, narrow regions and multiple headers. Unfortunately, on these devices, the regions with local high wall shear stresses (WSS) also show high local fouling rates. Several explanations have been put forward by the scientific community, including the effect of cell transport by Brownian motion on the adhesion rate. In this work, for the first time, both WSS and convection and Brownian diffusion effects on cell adhesion were evaluated along a microchannel with intercalate constriction and expansion zones designed to mimic the hydrodynamics of the human body and biomedical devices. Convection and Brownian diffusion effects were numerically studied using a steady-state convective-diffusion model (convection, diffusion and sedimentation). According to the numerical results, the convection and Brownian diffusion effects on cell adhesion are effectively more significant in regions with high WSS. Furthermore, a good agreement was observed between experimental and predicted local Sherwood numbers, particularly at the entrance and within the multiple constrictions. However, further mechanisms should be considered to accurately predict cell adhesion in the expansion zones. The described numerical approach can be used as a way to identify possible clogging zones in microchannels, and defining solutions, even before the construction of the prototype.

Fabrication and Hydrodynamic Characterization of a Microfluidic Device for Cell Adhesion Tests in Polymeric Surfaces.

A fabrication method is developed to produce a microfluidic device to test cell adhesion to polymeric materials. The process is able to produce channels with walls of any spin coatable polymer. The method is a modification of the existing poly-dimethylsiloxane soft lithography method and, therefore, it is compatible with sealing methods and equipment of most microfluidic laboratories. The molds are produced by xurography, simplifying the fabrication in laboratories without sophisticated equipment for photolithography. The fabrication method is tested by determining the effective differences in bacterial adhesion in five different materials. These materials have different surface hydrophobicities and charges. The major drawback of the method is the location of the region of interest in a lowered surface. It is demonstrated by bacterial adhesion experiments that this drawback has a negligible effect on adhesion. The flow in the device was characterized by computational fluid dynamics and it was shown that shear stress in the region of interest can be calculated by numerical methods and by an analytical equation for rectangular channels. The device is therefore validated for adhesion tests.

Numerical study on the hemodynamics of patient-specific carotid bifurcation using a new mesh approach.

The definition of a suitable mesh to simulate blood flow in the human carotid bifurcation has been investigated. In this research, a novel mesh generation method is proposed: hexahedral cells at the center of the vessel and a fine grid of tetrahedral cells near the artery wall, in order to correctly simulate the large blood velocity gradients associated with specific locations. The selected numerical examples to show the pertinence of the novel generation method are supported by carotid ultrasound image data of a patient-specific case. Doppler systolic blood velocities measured during ultrasound examination are compared with simulated velocities using 4 different combinations of hexahedral and tetrahedral meshes and different fluid dynamic simulators. The Lin's test was applied to show the concordance of the results. Wall shear stress-based descriptors and localized normalized helicity descriptor emphasize the performance of the new method. Another feature is the reduced computation time required by the developed methodology. With the accurate combined mesh, different flow rate partitions, between the internal carotid artery and external carotid artery, were studied. The overall effect of the partitions is mainly in the blood flow patterns and in the hot-spot modulation of atherosclerosis-susceptible regions, rather than in their distribution along the bifurcation.

Genetic diversity of Casearia sylvestris populations in remnants of the Atlantic Forest.

Guaçatonga (Casearia sylvestris) is a native plant of the Atlantic Forest, with high medicinal potential and relevance for reforestation programs. The aim of this study was to characterize, with microsatellite markers, two populations of C. sylvestris from remaining areas of the Atlantic Forest in the State of São Paulo. High allelic variation was found in both populations (NA = 101 and 117; AR = 12.5 and 14.4), although with high endogamy coefficients (f = 0.640 and 0.363). Estimates of genetic structure suggested the presence of considerable genetic divergence between the populations (FST = 0.103); however, there was no spatial genetic structure within the populations. Genetic divergence may have occurred due to decreased gene flow between the fragmented populations as the result of deforestation. The results of this study demonstrate the importance of genetic diversity and its characterization in native plants within remaining forest areas for the management and restoration of such areas.

Genetic diversity of Lippia sidoides Cham. and L. gracilis Schauer germplasm.

The conservation of plants in germplasm banks ensures the characterization and availability of these resources for future generations. The present study used DNA markers to obtain genetic information about germplasm collections of Lippia sidoides and L. gracilis, which are maintained in an Active Germplasm Bank (AGB). Genetic variability of samples in the AGB was assessed using 12 combinations of amplified fragment length polymorphism (AFLP) primers (EcoRI/MseI). Twenty simple sequence repeat primers designed for L. alba were tested to determine their transferability in L. sidoides and L. gracilis. The AFLP markers generated 789 markers. The assessed loci exhibited a moderate Shannon diversity index (I = 0.42) in both species, suggesting that the conserved accessions possess an intermediate level of genetic diversity. Twelve microsatellite loci amplified satisfactorily, and nine loci were polymorphic in each species. A total of 23, 22, and 36 alleles, with an average of 2.5, 2.4, and 3.27 alleles per locus were identified for L. sidoides and L. gracilis accessions in the AGB, and Lippia sp sampled plants, respectively. Analyses of genetic structure permitted the identification of three different groups using both sets of markers, of which two were representative of L. sidoides. The information generated in this study may help to create, expand, and maintain collections of these species and may assist in genetic-breeding programs.

Cross-species amplification and polymorphism of microsatellite loci in Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) in Brazilian cropping systems.

The Old World bollworm Helicoverpa armigera (Hübner) was recently discovered in Brazil. This species is closely related to the New World bollworm H. zea (Boddie), and mating between these species has already been reported under laboratory conditions. Here, we tested the cross-species amplification of 20 microsatellite (SSR) loci in field populations of H. armigera and H. zea collected from Brazilian cropping systems. Seven SSR loci were successfully amplified and polymorphic in both species except for the locus HaC14, which was monomorphic for H. zea. All SSR loci were in linkage equilibrium, and deviations from Hardy- Weinberg equilibrium were only observed for the locus HarSSR1 in the HaRS-2 population, where null alleles were present. A moderate level of polymorphism was detected in H. armigera and H. zea populations with a mean allele number of 4.14, and 2.24, respectively. Interestingly, most of the populations of the recent invader H. armigera showed higher genetic diversity and inbreeding coefficients than H. zea populations. The genetic identity of each species was recovered using a STRUCTURE analysis, where the populations formed two clusters (K = 2) according to their species. STRUCTURE also suggested the occurrence of potential hybrid offspring between H. armigera and H. zea individuals in natural conditions. These SSR loci will be valuable in characterizing population differentiation, invasion routes, adaptation, reproductive behavior, and intra- and interspecific gene flow in H. armigera and H. zea populations in Brazil, the USA, and other areas where these two pests occur.

Link between deviations from Murray's Law and occurrence of low wall shear stress regions in the left coronary artery.

Murray developed two laws for the geometry of bifurcations in the circulatory system. Based on the principle of energy minimization, Murray found restrictions for the relation between the diameters and also between the angles of the branches. It is known that bifurcations are prone to the development of atherosclerosis, in regions associated to low wall shear stresses (WSS) and high oscillatory shear index (OSI). These indicators (size of low WSS regions, size of high OSI regions and size of high helicity regions) were evaluated in this work. All of them were normalized by the size of the outflow branches. The relation between Murray's laws and the size of low WSS regions was analysed in detail. It was found that the main factor leading to large regions of low WSS is the so called expansion ratio, a relation between the cross section areas of the outflow branches and the cross section area of the main branch. Large regions of low WSS appear for high expansion ratios. Furthermore, the size of low WSS regions is independent of the ratio between the diameters of the outflow branches. Since the expansion ratio in bifurcations following Murray's law is kept in a small range (1 and 1.25), all of them have regions of low WSS with similar size. However, the expansion ratio is not small enough to completely prevent regions with low WSS values and, therefore, Murray's law does not lead to atherosclerosis minimization. A study on the effect of the angulation of the bifurcation suggests that the Murray's law for the angles does not minimize the size of low WSS regions.

QTL mapping of soybean oil content for marker-assisted selection in plant breeding program.

The present study was undertaken to detect and map the quantitative trait loci (QTL) related to soybean oil content. We used 244 progenies derived from a bi-parental cross of the Lineage 69 (from Universidade Estadual Paulista "Júlio de Mesquita Filho"/Faculdade de Ciências Agrárias e Veterinárias - Breeding Program) and Tucunaré cultivar. A total of 358 simple sequence repeat (SSR; microsatellite) markers were used to investigate the polymorphism between the parental lines, and for the polymorphic lines all the F2 individuals were tested. Evaluation of the oil content and phenotype was performed with the aid of a Tango equipment by near infra-red reflectance spectroscopy, using single F2 seeds and F2:3 progenies, in triplicate. The data were analyzed by QTL Cartographer program for 56 SSR polymorphic markers. Two oil-content related QTLs were detected on K and H linkage groups. The total phenotypic variation explained by QTLs ranged from 7.8 to 46.75% for oil content. New QTLs were identified for the oil content in addition to those previously identified in other studies. The results reported in this study show that regions different from those already known could be involved in the genetic control of soybean oil content.

Numerical study of wall shear stress-based descriptors in the human left coronary artery.

The present work is about the application of wall shear stress descriptors - time averaged wall shear stress (TAWSS), oscillating shear index (OSI) and relative residence time (RRT) - to the study of blood flow in the left coronary artery (LCA). These descriptors aid the prediction of disturbed flow conditions in the vessels and play a significant role in the detection of potential zones of atherosclerosis development. Hemodynamic descriptors data were obtained, numerically, through ANSYS® software, for the LCA of a patient-specific geometry and for a 3D idealized model. Comparing both cases, the results are coherent, in terms of location and magnitude. Low TAWSS, high OSI and high RRT values are observed in the bifurcation - potential zone of atherosclerosis appearance. The dissimilarities observed in the TAWSS values, considering blood as a Newtonian or non-Newtonian fluid, releases the importance of the correct blood rheologic caracterization. Moreover, for a higher Reynolds number, the TAWSS values decrease in the bifurcation and along the LAD branch, increasing the probability of plaques deposition. Furthermore, for a stenotic LCA model, very low TAWSS and high RRT values in front and behind the stenosis are observed, indicating the probable extension, in the flow direction, of the lesion.

Molecular and chemical characterization of vetiver, Chrysopogon zizanioides (L.) Roberty, germplasm.

Due to the economic interests in vetiver, Chrysopogon zizanioides (L.) Roberty, molecular and chemical studies are essential to generate information for its sustainable exploitation. The aim of this study was to undertake a molecular and chemical characterization of vetiver accessions of the active germplasm bank of the Universidade Federal de Sergipe. The molecular characteristics of the accessions were studied using amplified fragment length polymorphism markers, with a total of 14 primer combinations that generated 442 loci, allowing us to observe that these accessions have similar genomes. The vetiver accessions were divided into three distinct groups, where accession UFS-VET005 was the most differentiated and accession UFS-VET004 had the lowest essential oil content (0.70%). The content of the chemical constituents of the essential oils was observed to vary, with a predominance of khusimol, which ranged from 18.97 to 25.02%. It was possible to divide the vetiver accessions into two groups based on chemical composition, and these groups do not correlate with the molecular grouping. Therefore, it is necessary to perform molecular and chemical analyses to characterize vetiver accessions.

Development of a novel set of microsatellite markers for Lippia alba (Verbenaceae).

Microsatellite primers were developed and optimized for Lippia alba to characterize the L. alba germplasm bank of Universidade de São Paulo. A genomic library enabled the design of 9 microsatellite primers. Six of the 9 primers yielded polymorphic products, which defined 2 groups in the bank. The data provide support to characterize germplasm banks, genetic breeding programs for L. alba, and other genetic diversity studies and classifications of species in the genus Lippia.

Cross-species transferability of microsatellite markers in the genus Lippia.

The cross-species transferability of 20 microsatellite markers was tested in the genus Lippia. Eleven markers were polymorphic after screening 19 accessions of Lippia sidoides and Lippia gracilis maintained in the Active Germplasm Bank (AGB) from Universidade Federal de Sergipe. Additionally, 40 accessions of Lippia spp were collected in Sergipe to increase the germplasm bank. A total of 23, 22, and 36 alleles were identified, with an average of 2.3, 2.2, and 3.27 alleles per locus, respectively, for each group. The markers that were used were efficient tools to access genetic diversity in the germplasm bank and will be useful for further research aiming at the conservation and management of these important aromatic species.