Mechanistic Studies on the Self-Assembly of PLGA Patchy Particles and Their Potential Applications in Biomedical Imaging.

Currently, several challenges prevent poly(lactic-co-glycolic acid) (PLGA) particles from reaching clinical settings. Among these is a lack of understanding of the molecular mechanisms involved in the formation of these particles. We have been studying in depth the formation of patchy polymeric particles. These particles are made of PLGA and lipid-polymer functional groups. They have unique patch-core-shell structural features: hollow or solid hydrophobic cores and a patchy surface. Previously, we identified the shear stress as the most important parameter in a patchy particle's formation. Here, we investigated in detail the role of shear stress in the patchy particle's internal and external structure using an integrative experimental and computational approach. By cross-sectioning the multipatch particles, we found lipid-based structures embedded in the entire PLGA matrix, which represents a unique finding in the PLGA field. By developing novel computational fluid dynamics and molecular dynamics simulations, we found that the shear stress determines the internal structure of the patchy particles. Equally important, we discovered that these particles emit a photoacoustic (PA) signal in the optical clinical imaging window. Our results show that particles with multiple patches emit a higher PA signal than single-patch particles. This phenomenon most likely is due to the fact that multipatchy particles absorb more heat than single-patchy particles as shown by differential scanning calorimetry analysis. Furthermore, we demonstrated the use of patchy polymeric particles as photoacoustic molecular probes both in vitro and in vivo studies. The fundamental studies described here will help us to design more effective PLGA carriers for a number of medical applications as well as to accelerate their medical translation.

Strategy for modeling flow diverters in cerebral aneurysms as a porous medium.

Simulations using the patient-specific geometry of the aneurysm may help in a better planning of the treatment and in a consequent reduction of the associated risks. We propose, evaluate, and implement a methodology for the simulation of flow diverter (FD) devices in intracranial aneurysms by using a porous medium method (PMM), which greatly reduces the computational cost of these simulations compared with immersed method (IMM) approaches used to model complex FDs. The method relies on parameters from an empirical correlation derived from experimental observations in wire screens, consistent with CFD simulations. The verification of our PMM strategy was carried out by comparing the results of simulations in different (patient-specific) geometries and FDs, to those obtained under identical conditions by the IMM. Overall, both quantitative and qualitative results are consistent between IMM and PMM in cases where the local porosity remains roughly uniform throughout the neck, with differences in the reduction of the observables lower than 10%. This PMM strategy is up to 10 times faster than the IMM, which allows for a runtime of hours instead of days, bringing it closer for its application in the clinic.

Clinical Application of Image-Based CFD for Cerebral Aneurysms.

During the last decade, the convergence of medical imaging and computational modeling technologies has enabled tremendous progress in the development and application of image-based computational fluid dynamics modeling of patient-specific blood flows. These techniques have been used for studying the basic mechanisms involved in the initiation and progression of vascular diseases, for studying possible ways to improve the diagnosis and evaluation of patients by incorporating hemodynamics information to the anatomical data typically available, and for the development of computational tools that can be used to improve surgical and endovascular treatment planning. However, before these technologies can have a significant impact on the routine clinical practice, it is still necessary to demonstrate the connection between the extra information provided by the models and the natural progression of vascular diseases and the outcome of interventions. This paper summarizes some of our contributions in this direction, focusing in particular on cerebral aneurysms.

Signatures of vicariance, postglacial dispersal and spawning philopatry: population genetics of the walleye Sander vitreus.

Population genetic relationships reveal the signatures of current processes such as reproductive behaviour and migration, as well as historic events including vicariance and climate change. We analyse population structure of native walleye Sander vitreus across North America, encompassing 10 nuclear DNA microsatellite loci, 26 spawning sites and 921 samples from watersheds across the Great Lakes, Lake Winnipeg, upper Mississippi River, Ohio River and Mobile Bay of the Gulf Coast. Geographical patterning is assessed using phylogenetic trees, pairwise F(ST) analogues, hierarchical partitioning, Mantel regression, Bayesian assignment and Monmonier geographical networks. Results reveal congruent divergences among population groups, corresponding to historic isolation in glacial refugia, dispersal patterns and basin divisions. Broad-scale relationships show genetic isolation with geographical distance, but reproductive groups within basins do not -- with some having pronounced differences. Greatest divergence distinguishes outlying Gulf Coastal and northwest populations, the latter tracing to dispersal from the Missourian refugium to former glacial Lake Agassiz, and basin isolation approximately 7000 ya. Genetic barriers in the Great Lakes separate groups in Lakes Superior, Huron's Georgian Bay, Erie and Ontario, reflecting contributions from Mississippian and Atlantic refugia, and changes in connectivity patterns. Walleye genetic patterns thus reflect vicariance among watersheds and glacial refugia, followed by re-colonization pathways and changing drainage connections that established modern-day northern populations, whose separations are maintained through spawning site fidelity. Conservation management practices should preserve genetic identity and unique characters among these divergent walleye populations.

Merging of intersecting triangulations for finite element modeling.

Surface mesh generation over intersecting triangulations is a problem common to many branches of biomechanics. A new strategy for merging intersecting triangulations is described. The basis of the method is that object surfaces are represented as the zero-level iso-surface of the distance-to-surface function defined on a background grid. Thus, the triangulation of intersecting objects reduces to the extraction of an iso-surface from an unstructured grid. In a first step, a regular background mesh is constructed. For each point of the background grid, the closest distance to the surface of each object is computed. Background points are then classified as external or internal by checking the direction of the surface normal at the closest location and assigned a positive or negative distance, respectively. Finally, the zero-level iso-surface is constructed. This is the final triangulation of the intersecting objects. The overall accuracy is enhanced by adaptive refinement of the background grid elements. The resulting surface models are used as support surfaces to generate three-dimensional grids for finite element analysis. The algorithms are demonstrated by merging arterial branches independently reconstructed from contrast-enhanced magnetic resonance images and by adding extra features such as vascular stents. Although the methodology is presented in the context of finite element analysis of blood flow, the algorithms are general and can be applied in other areas as well.

Expression of human milk fat globulin proteins in cells of haemopoietic origin.

Lineage-specific gene expression has been used for the identification of metastasis of cancers with unknown primary site or of disseminated cancer cells in haemopoietic compartments such as bone marrow or in lymph nodes. For the muc1, cytokeratin-19 and the CEA genes, the transcription in haemopoietic cells has been shown recently. Here, the expression of the mammary epithelium related antigens BA46 (lactadherin) and BA70 in lymphoid and myeloid cell lines, and in clinical specimens is analysed. By Northern-hybridization with specific oligonucleotides an ubiquitous transcription of both genes, independent from the provenance of cells or the chromosomal gender was found. Both mRNA molecules were amplified by rtPCR from the samples and the specificity could be confirmed by sequence analysis. Peptide-specific antibodies were raised in rabbits and used for Western-blot analysis and for immunocytochemical studies. Both antibodies reacted with total cell lysates from myeloid and lymphatic cells. In immunocytochemistry antibody P717 (anti-lactadherin) had a significant strong staining of the myeloid cell lines K562 and HL60 suggesting a participation of lactadherin in leukocyte-function. Using antibody P718, strong stains were seen in myeloid line K562 and lymphoid line ST486. In conclusion, our findings expand the results that the concept of lineage-specific gene expression is no longer valid at the molecular level.

Improving acute skin-flap survival through stress conditioning using heat shock and recovery.

We present our initial experience with a new method of increasing the survival of acute skin flaps through stress conditioning using heat shock and recovery. The heat-shock response is a basic form of stress response that exists on the cellular level. When cultured cells or whole organisms are exposed to supraphysiologic levels of heat, they respond by synthesizing a number of highly conserved proteins known as heat-shock proteins. These proteins have been shown to offer the cell or organism a survival advantage over nonstressed controls. The study demonstrates a significant survival advantage in acute dorsal skin flaps of Sprague-Dawley rats (p = 0.001). Study animals (n = 10) were subjected to a heating blanket set at 45 degrees C for 30 minutes and were allowed 6 hours' recovery before developing the flaps. Heat-shock protein was demonstrated in immunohistochemically stained sections of skin from the study animals but not in control animal skin (n = 14). We postulate that through stress conditioning a latent mechanism present within all cells was activated, thereby allowing the cells of our experimental flaps to better survive the stress of the acute flap model.