New approaches to biomaterials design.

The adoption of combinatorial and computational methods in biomaterials design is a highway towards the discovery and realization of tailored polymeric materials to satisfy the specific requirements of many diverse biomedical or prosthetic applications.

Random-coil behavior and the dimensions of chemically unfolded proteins.

Spectroscopic studies have identified a number of proteins that appear to retain significant residual structure under even strongly denaturing conditions. Intrinsic viscosity, hydrodynamic radii, and small-angle x-ray scattering studies, in contrast, indicate that the dimensions of most chemically denatured proteins scale with polypeptide length by means of the power-law relationship expected for random-coil behavior. Here we further explore this discrepancy by expanding the length range of characterized denatured-state radii of gyration (R(G)) and by reexamining proteins that reportedly do not fit the expected dimensional scaling.

A photo-crosslinked poly(vinyl alcohol) hydrogel growth factor release vehicle for wound healing applications.

The objective of this study was to develop and evaluate a hydrogel vehicle for sustained release of growth factors for wound healing applications. Hydrogels were fabricated using ultraviolet photo-crosslinking of acrylamide-functionalized nondegradable poly(vinyl alcohol) (PVA). Protein permeability was initially assessed using trypsin inhibitor (TI), a 21 000 MW model protein drug.

Using surrogate modeling in the prediction of fibrinogen adsorption onto polymer surfaces.

We present a Surrogate (semiempirical) Model for prediction of protein adsorption onto the surfaces of biodegradable polymers that have been designed for tissue engineering applications. The protein used in these studies, fibrinogen, is known to play a key role in blood clotting. Therefore, fibrinogen adsorption dictates the performance of implants exposed to blood.

Preliminary development of a novel resorbable synthetic polymer fiber scaffold for anterior cruciate ligament reconstruction.

We are developing novel resorbable fiber-based scaffolds for reconstruction of the anterior cruciate ligament (ACL). For the first time, we report fabrication of fibers from poly(DTE carbonate) polymer. Poly(L-lactic acid) fibers were also fabricated for comparison purposes.

Poly(ethylene glycol) enhances cell motility on protein-based poly(ethylene glycol)-polycarbonate substrates: a mechanism for cell-guided ligand remodeling.

The regulation of cell motility on ligand-adsorbed poly(ethylene glycol) (PEG)-based polymeric biomaterials is governed by variables that are not well characterized. In this report, we examined keratinocyte migratory responsiveness to PEG-variant tyrosine-derived polycarbonates adsorbed with equivalent levels of the cell adhesion ligand, fibronectin. The equivalently adsorbed ligand adopted differential distributions, confirmed via atomic force microscopy, and the total number of exposed cell-binding domains (CBD), quantified through immunosorbent fluorometry, varied as a function of PEG concentration.

Small changes in the polymer structure influence the adsorption behavior of fibrinogen on polymer surfaces: validation of a new rapid screening technique.

Numerous studies conclude that the selective adsorption of plasma proteins on materials contacting blood or tissue affects all subsequent interactions related to the biocompatibility of artificial surfaces. However, there are only a few studies available, which clearly demonstrate that there is a correlation between surface chemistry and selective protein adsorption. Detailed knowledge of such correlations would facilitate the design of biocompatible materials.

Association of hydrophobically-modified poly(ethylene glycol) with fusogenic liposomes.

We present results on using cooperative interactions to shield liposomes by incorporating multiple hydrophobic anchoring sites on polyethylene glycol (PEG) polymers. The hydrophobically-modified PEGs (HMPEGs) are comb-graft polymers with strictly alternating monodisperse PEG blocks (M(w)=6, 12, or 35 kDa) bonded to C18 stearylamide hydrophobes. Cooperativity is varied by changing the degree of oligomerization at a constant ratio of PEG to stearylamide.

Polymers derived from the amino acid L-tyrosine: polycarbonates, polyarylates and copolymers with poly(ethylene glycol).

The natural amino acid L-tyrosine is a major nutrient having a phenolic hydroxyl group. This feature makes it possible to use derivatives of tyrosine dipeptide as a motif to generate diphenolic monomers, which are important building blocks for the design of biodegradable polymers. Particularly useful monomers are desaminotyrosyl-tyrosine alkyl esters (abbreviated as DTR, where R stands for the specific alkyl ester used).

Hummingbird foraging position is altered by the touch-sensitive stigma of bush monkeyflower.

The two stigma lobes of bush monkeyflower (Mimulus aurantiacus) close together rapidly in response to touch by a hummingbird pollinator and usually remain closed for the life of the flower, preventing further pollen receipt. Previous work showed that hummingbirds visiting bush monkeyflowers with closed stigmas export more than twice as much pollen to recipient flowers as birds visiting flowers with open stigmas. To investigate how stigma closure increases pollen export, we used videotape to examine the interaction between bird and flower.

Electrostatic contributions to site specific DNA cleavage by EcoRV endonuclease.

Mutational analysis of amino acids at the periphery of the EcoRV endonuclease active site suggests that moderate-range electrostatic effects play a significant role in modulating the efficiency of phosphoryl transfer. Asp36 and Lys38 located on minor-groove binding surface loops approach within 7-9 A of the scissile phosphates of the DNA. While the rates of single-site mutations removing the carboxylate or amine moieties at these positions are decreased 10(3)-10(5)-fold compared to that of wild-type EcoRV, we find that double mutants which rebalance the charge improve catalysis by up to 500-fold.

S-allele diversity in Sorbus aucuparia and Crataegus monogyna (Rosaceae: Maloideae).

RT-PCR was used to obtain the first estimates from natural populations of allelic diversity at the RNase-based gametophytic self-incompatibility locus in the Rosaceae. A total of 20 alleles were retrieved from 20 Sorbus aucuparia individuals, whereas 17 alleles were found in 13 Crataegus monogyna samples. Estimates of population-level allele numbers fall within the range observed in the Solanaceae, the only other family with RNase-based incompatibility for which estimates are available.

A randomized clinical trial of the effectiveness of an acupressure device (relief brief) for managing symptoms of dysmenorrhea.

Objectives: To develop and test the safety and effectiveness of an acupressure garment (the Relief Brief) in decreasing the pain and symptom distress associated with dysmenorrhea.

Design: A randomized clinical trial applied a 2 (Relief Brief use or control group) x 3 (baseline and two treatment measurement occasions) mixed factorial design.

Participants: Sixty-one (61) women with moderately severe primary dysmenorrhea were randomly assigned to the standard treatment control group or the Relief Brief acupressure device group after one pretreatment menses, with 58 women reporting the effect on their pain during two post-treatment menstrual cycles.

Biomaterials and scaffolds in reparative medicine.

Most approaches currently pursued or contemplated within the framework of reparative medicine, including cell-based therapies, artificial organs, and engineered living tissues, are dependent on our ability to synthesize or otherwise generate novel materials, fabricate or assemble materials into appropriate 2-D and 3-D forms, and precisely tailor material-related physical and biological properties so as to achieve a desired clinical response. This paper summarizes the scientific and technological opportunities within the fields of biomaterials science and molecular engineering that will likely establish new enabling technologies for cellular and molecular therapies directed at the repair, replacement, or reconstruction of diseased or damaged organs and tissues.

Phylogenetic congruence and discordance among one morphological and three molecular data sets from Pontederiaceae.

A morphological data set and three sources of data from the chloroplast genome (two genes and a restriction site survey) were used to reconstruct the phylogenetic history of the pickerelweed family Pontederiaceae. The chloroplast data converged towards a single tree, presumably the true chloroplast phylogeny of the family. Unrooted trees estimated from each of the three chloroplast data sets were identical or extremely similar in shape to each other and mostly robustly supported.

Development of photo-crosslinking reagents for protein kinase-substrate interactions.

The identification of relevant protein kinase-protein substrate partners remains a serious challenge on a genome-wide scale. The design and synthesis of a photo-activatable nucleotide reagent to crosslink protein kinases with their substrates is described in which an azido group is appended to the gamma-phosphoryl and purine moieties of ATP. In the absence of UV, compounds of this class were shown to act as competitive inhibitors versus ATP and non-competitive inhibitors versus peptide substrate for the protein tyrosine kinase Csk, suggesting that they can form a ternary complex with kinase and protein substrate.

A synthetic polymer matrix for the delayed or pulsatile release of water-soluble peptides.

The design of a polymeric peptide release system with a controlled delay time and a burst-free pre-release phase is described. In general, the system consists of a blend of a tyrosine-derived polyarylate and a fast-degrading copolymer of lactic and glycolic acid (PLGA). Due to the peptide-like structure of the polyarylate backbone, peptide-polymer interactions prevented the release of peptide from neat polyarylate films.

CTLA-4/CD 28 region polymorphisms in children from families with autoimmune hepatitis.

Susceptibility to autoimmune hepatitis is associated with particular human leucocyte antigen class II alleles. However, non-HLA genetic factors are likely to be required for development of the disease. Among the candidate genes, the cytotoxic T-lymphocyte antigen 4 (CTLA-4) and CD28 genes, located on chromosome 2q33 in humans, encode a cell surface molecule playing a dominant role in the regulation of T-cell activation.

Evolutionary relationships among self-incompatibility RNases.

T2-type RNases are responsible for self-pollen recognition and rejection in three distantly related families of flowering plants-the Solanaceae, Scrophulariaceae, and Rosaceae. We used phylogenetic analyses of 67 T2-type RNases together with information on intron number and position to determine whether the use of RNases for self-incompatibility in these families is homologous or convergent. All methods of phylogenetic reconstruction as well as patterns of variation in intron structure find that all self-incompatibility RNases along with non-S genes from only two taxa form a monophyletic clade.

Tissue spreading on implantable substrates is a competitive outcome of cell-cell vs. cell-substratum adhesivity.

While the interactions of cells with polymeric substrata are widely studied, the influence of cell-cell cohesiveness on tissue spreading has not been rigorously investigated. Here we demonstrate that the rate of tissue spreading over a two-dimensional substratum reflects a competition or "tug-of-war" between cell-cell and cell-substratum adhesions. We have generated both a "library" of structurally related copolymeric substrata varying in their adhesivity to cells and a library of genetically engineered cell populations varying only in cohesiveness.

The association of coagulase-negative staphylococci isolated from the chorioamnion at delivery and subsequent development of cerebral palsy.

Objective: To find out whether there is an association between cultures positive for coagulase negative staphylococci (CONS) taken from babies in the Neonatal Intensive Care Unit (NICU) and a subsequent outcome of cerebral palsy.

Study Design: At delivery, we obtained cultures from the chorioamnion space and, when medically indicated, we obtained bacterial cultures from children in the NICU. Surviving neonates underwent final examination for cerebral palsy at age 18 months.

Hydrolytic degradation of tyrosine-derived polycarbonates, a class of new biomaterials. Part II: 3-yr study of polymeric devices.

The kinetics and mechanisms of in vitro degradation of tyrosine-derived polycarbonates, a new class of polymeric biomaterials, were studied extensively at 37 degrees C. These polymers carry an alkyl ester pendent chain that allows the fine-tuning of the polymer's material properties, its biological interactions with cells and tissue, and its degradation behavior. The polymer carrying an ethyl ester pendent chain, poly(DTE carbonate), has been established as a promising orthopedic implant material, exhibiting bone apposition when in contact with hard tissue.