A Variable Neighbourhood Descent Heuristic for Conformational Search Using a Quantum Annealer.

Discovering the low-energy conformations of a molecule is of great interest to computational chemists, with applications in in silico materials design and drug discovery. In this paper, we propose a variable neighbourhood search heuristic for the conformational search problem. Using the structure of a molecule, neighbourhoods are chosen to allow for the efficient use of a binary quadratic optimizer for conformational search. The method is flexible with respect to the choice of molecular force field and the number of discretization levels in the search space, and can be further generalized to take advantage of higher-order binary polynomial optimizers. It is well-suited for the use of devices such as quantum annealers. After carefully defining neighbourhoods, the method easily adapts to the size and topology of these devices, allowing for seamless scaling alongside their future improvements.

Complement C5 Inhibition Reduces T Cell-Mediated Allograft Vasculopathy Caused by Both Alloantibody and Ischemia Reperfusion Injury in Humanized Mice.

Allograft vasculopathy (AV) is characterized by diffuse stenoses in the vasculature of solid organ transplants. Previously, we developed two humanized models showing that alloantibody and ischemia reperfusion injury (IRI) exacerbated T cell-mediated AV in human arterial xenografts in vivo. Herein we examined a causal role for terminal complement activation in both settings. IRI, in contrast to alloantibody, elicited widespread membrane attack complex (MAC) assembly throughout the vessel wall. Both alloantibody and IRI caused early (24 h) and robust endothelial cell (EC) activation localized to regions of intimal MAC deposition, indicated by increases in nuclear factor kappa B (NF-κB)-inducing kinase, an MAC-dependent activator of noncanonical NF-kB, VCAM-1 expression and Gr-1+ neutrophil infiltration. Endothelial cell activation by alloantibody was inhibited by antimouse C5 mAb, but not by anti-C5a mAb or by control mAb, implicating MAC as the primary target of anti-C5 mAb. Antimouse C5 mAb significantly reduced alloantibody- and IRI-enhanced T cell infiltration and AV-like changes, including neointimal hyperplasia as well as intraluminal thrombosis in a subset of IRI-treated arterial grafts. These results indicate that increased AV lesion formation in response to either alloantibody or IRI is dependent on complement C5 activation and, accordingly, inhibition of this pathway may attenuate AV.

Tyrosine-derived polycarbonate-silica xerogel nanocomposites for controlled drug delivery.

Biodegradable polymer-ceramic composites offer significant potential advantages in biomedical applications where the properties of either polymers or ceramics alone are insufficient to meet performance requirements. Here we demonstrate the highly tunable mechanical and controlled drug delivery properties accessible with novel biodegradable nanocomposites prepared by non-covalent binding of silica xerogels and co-polymers of tyrosine-poly(ethylene glycol)-derived poly(ether carbonate). The Young's moduli of the nanocomposites exceed by factors of 5-20 times those of the co-polymers or of composites made with micron scale silica particles. Increasing the fraction of xerogel in the nanocomposites increases the glass transition temperature and the mechanical strength, but decreases the equilibrium water content, which are all indicative of strong non-covalent interfacial interactions between the co-polymers and the silica nanoparticles. Sustained, tunable controlled release of both hydrophilic and hydrophobic therapeutic agents from the nanocomposites is demonstrated with two clinically significant drugs, rifampicin and bupivacaine. Bupivacaine exhibits an initial small burst release followed by slow release over the 7 day test period. Rifampicin release fits the diffusion-controlled Higuchi model and the amount released exceeds the dosage required for treatment of clinically challenging infections. These nanocomposites are thus attractive biomaterials for applications such as wound dressings, tissue engineering substrates and stents.

Tyrosine-derived nanospheres for enhanced topical skin penetration.

The objective of this study was to investigate the passive skin penetration of lipophilic model agents encapsulated within tyrosine-derived nanospheres. The nanospheres were formed by the self-assembly of a biodegradable, non-cytotoxic ABA triblock copolymer. The A-blocks were poly(ethylene glycol) and the hydrophobic B-blocks were oligomers of suberic acid and desaminotyrosyl-tyrosine alkyl esters. These nanospheres had an average hydrodynamic diameter of about 50nm and formed strong complexes with fluorescent dyes, 5-dodecanoylaminofluorescein (DAF, LogD=7.54) and Nile Red (NR, LogD=3.10). These dyes have been used here as models for lipophilic drugs. The distribution of topically applied nanosphere-dye formulations was studied in human cadaver skin using cryosectioning and fluorescence microscopy. Permeation analysis (quantified fluorescence) over a 24h period revealed that the nanospheres delivered nine times more NR to the lower dermis than a control formulation using propylene glycol. For DAF, the nanosphere formulation was 2.5 times more effective than the propylene glycol based control formulation. We conclude that tyrosine-derived nanospheres facilitate the transport of lipophilic substances to deeper layers of the skin, and hence may be useful in topical delivery applications.

Acute tensile strength analysis of collagen solder for mesh fixation to the peritoneal surface.

BACKGROUND: In this study, we assessed the feasibility of laser-assisted tissue welding as a means of fixing mesh prostheses to the peritoneum. We then tested the initial tensile strength of the bonds. METHODS: Fresh porcine peritoneal coupons were lap-joint bonded with laser-activated solder. Anesthetized New Zealand white rabbits and Yorkshire pigs also underwent laparotomy. Vicryl mesh (2.0 x 1.0 cm) was attached to the peritoneum using a laser system (1.43 mu, 2.5 W, 60 degrees C), solder formulations and configurations, and a 1 cm2 bond area. Control segments were affixed with 4.8-mm staples. After the animals were killed, the segments were excised en bloc. Tensile strength assessment was conducted by measuring peak force breaking strength. RESULTS: The strength of the solder bonds were similar in all groups (range, 261.5 +/- 170.3-465.3 +/- 288.2 g/cm2) and were not statistically different from the controls (215.8 +/- 117.8 g/cm2). CONCLUSIONS: These values are similar to the 200-500 g/cm2 acute strengths reported for sutured or stapled peritoneal closure. Mesh fixation by solder is feasible, and further development of this technology is warranted.

Repair of a critical size defect in the rat mandible using allogenic type I collagen.

Mandibular fractures, resulting from either trauma or reconstructive surgery, can be challenging craniofacial problems. The morbidity of failed fracture healing is significant and may require bone grafting. Donor site morbidity and finite amounts of autogenous bone are major drawbacks of autogenous bone grafting. Similarly, the use of allografts and xenografts may be associated with an increased risk of rejection, infection, and nonunion. To circumvent the limitations of bone grafting, research efforts have focused on formulating a suitable bone substitute. The purpose of our study was to evaluate the efficacy of type I collagen implants in repairing critical sized mandibular defects in rats. Twelve male Sprague-Dawley rats (200-300g) were divided equally into control and experimental groups. Full thickness, round, four millimeter in diameter defects were created in the ramus of the right mandible of all rats using an electrical burr at low speed. The defects were irrigated of all bone chips, and either filled with a precisely fitted disk of allogenic collagen type I gel (experimental animals) or left empty (control animals). Animals were killed 6 weeks after surgery and healing of the bone defects was assessed in a blinded fashion using radiologic and histologic analysis. Radiologic analysis of the control group revealed a clear circular right mandibular defect in all animals, whereas the collagen disk implant group revealed an indistinct to nonexistent right mandibular defect in all animals. Densitometric analysis revealed a significant difference between these groups (* P = 0.01). Similarly, gross analysis of control mandibles revealed a 4mm round, soft-tissue filled defect, while implanted defects demonstrated gross bone spanning the defect. Finally, histologic analysis of all control mandibles revealed clearly demarcated bony edges at the defect border with connective tissue spanning the defect. In contrast, histological analysis of all implanted mandibles revealed indistinct bony edges at the defect border with a thin layer of osteoblasts and viable bone spanning the defects. We have demonstrated the ability of type I collagen to promote healing of a membranous bony defect that would not otherwise heal at 6 weeks. The suitability of type I collagen as a carrier matrix provides ample opportunity for tissue-engineered approaches to further facilitate bony defect healing. Promoting bone formation through tissue engineering matrices offers great promise for skeletal healing and reconstruction.

Viscoelastic properties of human skin and processed dermis.

BACKGROUND/AIMS: The purpose of this work is to attempt to determine the elastic spring constant for collagen and elastic fibers (elastin) in skin and to determine if the values of these elastic constants are similar to those reported for other tissues. METHODS: We studied the viscoelastic mechanical properties of human skin and dermis by measuring the incremental stress-strain behavior. Elastic stress-strain curves were used to obtain the elastic spring constant of elastin and collagen while the collagen fibril length was obtained from the slope of viscous stress-strain curves. RESULTS: Our results suggest that the elastic spring constant for elastin is about 4.0 MPa while that for collagen is about 4.4 GPa. The former value is similar to that calculated for ligamentum nuchae while the latter value is about 70% of the value found for tendon and self-assembled type I collagen fibers. The differences between the elastic constants for collagen molecules in tendon and skin is hypothesized to reflect the higher molecular tilt angle and lower D period found in skin compared to tendon as well as a shorter fibril length. CONCLUSION: The differences in the collagen types present in skin and tendon may influence collagen self-assembly and the resulting viscoelastic properties.

The biological behavior of autologous collagen-based extracellular matrix injected into the rabbit bladder wall.

Endoscopic techniques are providing a minimally invasive approach to the treatment of vesicoureteral reflux and urinary incontinence. Bovine collagen has been used, but potential degradation over time and sensitivity reactions have limited its usefulness. We evaluate the use of an autologous collagen-based extracellular matrix preparation injected within the rabbit bladder submucosa and compare it to the stability of bovine collagen-injected similarly. Of 28 New Zealand white rabbits, 12 underwent injection of autologous collagen-based extracellular matrix, 12 bovine collagen, and four normal saline sham injections into the anterior bladder submucosa. Twelve collagen specimens were iodinated with iodine 125 ((125)I) paraaminobenzoate reagent. The (125)I-labeled rabbits were monitored with a gamma camera to assess the level of decay of radioactivity over 12 weeks. All animals were killed 12 weeks post-injection. Assessment of radioactivity showed minimal difference in radioisotope labeling between the autologous and bovine collagen (total counts and decay kinetics). Compared to bovine collagen, the autologous collagen- based extracellular matrix implants histologically showed increased fibroblastic and vascular infiltration focally. The most significant histologic difference was the marked inflammatory response associated with the bovine collagen implants. These data suggest that in the short term, autologous and bovine collagen appear to have similar stability. The response to autologous collagen-based extracellular matrix may increase longevity of the implant, primarily by reducing immunologic rejection and improving biocompatibility within the host tissue. Further long-term studies are necessary to assess the long-term stability of autologous collagen-based extracellular matrix. Neurourol. Urodynam. 18:487-495, 1999.

Identification of a (CUG)n triplet repeat RNA-binding protein and its expression in myotonic dystrophy.

Myotonic dystrophy (DM) is an autosomal dominant neuromuscular disease that is associated with a (CTG)n repeat expansion in the 3'-untranslated region of the myotonin protein kinase (Mt-PK) gene. This study reports the isolation and characterization of a (CUG)n triplet repeat pre-mRNA/mRNA binding protein that may play an important role in DM pathogenesis. Two HeLa cell proteins, CUG-BP1 and CUG-BP2, have been purified based upon their ability to bind specifically to (CUG)8 oligonucleotides in vitro. While CUG-BP1 is the major (CUG)8-binding activity in normal cells, nuclear CUG-BP2 binding activity increases in DM cells. Both CUG-BP1 and CUG-BP2 have been identified as isoforms of a novel heterogeneous nuclear ribonucleoprotein (hnRNP), hNab50. The CUG-BP/hNab50 protein is localized predominantly in the nucleus and is associated with polyadenylated RNAs in vivo. In vitro RNA-binding/photocrosslinking studies demonstrate that CUG-BP/hNab50 binds to RNAs containing the Mt-PK 3'-UTR. We propose that the (CUG)n repeat region in Mt-PK mRNA is a binding site for CUG-BP/hNab50 in vivo, and triplet repeat expansion leads to sequestration of this hnRNP on mutant Mt-PK transcripts.

An FGF receptor signaling pathway is required for the normal cell migrations of the sex myoblasts in C. elegans hermaphrodites.

The sex myoblasts (SMs) in C. elegans hermaphrodites undergo anteriorly directed cell migrations that allow for the proper localization of the egg-laying muscles. These migrations are controlled in part by a signal emanating from gonadal cells that allows the SMs to be attracted to their precise final positions flanking the center of the gonad. Mutations in egl-15 alter the nature of the interaction between the gonad and the SMs, resulting in the posterior displacement of the SMs. Here we show that egl-15 encodes a receptor tyrosine kinase of the fibroblast growth factor receptor (FGFR) subfamily with multiple roles in development. Three genes were identified that behave genetically as activators or mediators of egl-15 activity. One of these genes, sem-5, encodes an adaptor molecule that transduces signals from a variety of receptor tyrosine kinases. Like egl-15 and sem-5, the other two genes may similarly act in FGFR signaling pathways in C. elegans.

In vivo evaluation of a collagen corneal allograft derived from rabbit dermis.

BACKGROUND: This study evaluates epithelialization, clarity, intraocular inflammation, and fibroblast ingrowth of a collagen corneal allograft derived from rabbit dermis. METHODS: Dermal collagen fibers were dispersed intact and chemically modified to make them soluble. The allografts consisted of a fibrous, opaque peripheral zone and a central clear area. Twelve New Zealand white rabbits underwent penetrating keratoplasty with implantation of an allograft. The grafts were evaluated daily for clarify, anterior segment inflammation, and extent of reepithelialization with a slit-lamp microscope. RESULTS: Epithelialization of the fibrous peripheral zone of the graft ranged from 0% to 90%. The central clear area did not epithelialize in any of the animals. Fibroblasts migrated into the peripheral zone in all eyes. Complications included ulceration of the central clear area in two eyes. There was no ulceration or leakage at the graft-host interface and no synechia, fibrous, or inflammatory retrocorneal membranes in any of the eyes. CONCLUSIONS: Our study is the first to describe the method of modifying dermal type I collagen into a clear corneal allograft. Survival of the corneal collagen allograft beyond 1 month may be limited by several factors including lack of epithelialization of the central clear area.

The biological behavior of autologous collagen injected into the rabbit bladder.

Growing interest in the endoscopic treatment of vesicoureteral reflux and urinary incontinence has led to the development of safe reliable techniques that use an injectable bulking agent. A variety of injectable materials has been studied, such as polytetrafluoroethylene, fat and bovine collagen. We evaluated autologous collagen extracted from the dermis of skin for potential use in the urinary tract. Therefore, an animal model was developed to study the biological behavior of autologous collagen within the bladder. Collagen was extracted from the skin of rabbits and radiolabeled with 125iodine. An aliquot of radiolabeled autologous collagen was injected submucosally in the bladder. The animals were monitored weekly under a gamma camera. The decay of the radiolabeled collagen was recorded during 50 days. The animals were then sacrificed and the bladder wall at the site of injection was studied histologically and on electron microscopy. Decay of the injected collagen within the urinary tract was similar to control material, indicative of good stability within the bladder. A mild inflammatory response with fibroblastic and vascular invasion of the collagen implant was observed. These preliminary data suggest that autologous collagen injected endoscopically can provide a safe reliable bulking material that could be used to treat urinary incontinence and lower grades of vesicoureteral reflux in children.

Rapidly polymerized collagen gel as a smoothing agent in excimer laser photoablation.

BACKGROUND: Excimer laser photoablation effectively and precisely removes corneal tissue but may not smooth irregularities on the anterior corneal surface. An even surface might be obtained by applying a smoothing substance that fills in irregularities and ablates at the same rate as corneal tissue. Evaluation of collagen gel as a smoothing agent is reported. METHODS: Pure, type I collagen solutions were prepared to remain soluble at physiological pH and to spontaneously gel when exposed to cationic buffers. Collagen gels were formed on the surface of enucleated porcine corneas and on human donor corneas and exposed to varying pulses of 193-nanometer excimer laser energy. Effects of collagen gel on ablation depth, corneal surface smoothness, and smoothing of roughened cornea were evaluated by examining scanning electron micrographs of control and treated specimens. RESULTS: Collagen gels formed from 5 mg/ml collagen solutions dramatically altered the depth of photoablation. Photoablation of roughened cornea with collagen gels produced smooth corneal surfaces, whereas control eyes remained roughened and irregular. The smoothness of photoablated cornea surfaces improved when collagen gels were applied prior to photoablation. CONCLUSIONS: The rapidly gelling collagen solution appears to exhibit the functional properties required for an effective smoothing agent for excimer laser photoablation. The substance has low viscosity when applied to the corneal surface and forms a rigid gel when exposed to cationic buffer solution. In addition, the concentration of the collagen gel can be adjusted to provide different ablation rates.

Legal considerations for treatment following trauma to teeth.

When treating an orally traumatized patient, the dentist has no fewer professional and legal responsibilities than when treating a nonemergency patient. After obtaining proper consent, the examination, diagnosis, treatment, and follow-up of the patient must not be made negligently. Minimally, the dentist must not practice below the standard of care of a similar practitioner acting in the same or similar circumstances. The dentist also may be called on to further assist that patient in his or her attempt to obtain compensation for loss brought about by the trauma.

Extending and connecting signaling pathways in C. elegans.

The development of the nematode Caenorhabditis elegans is known to depend extensively on reproducible cell-cell interactions. The analysis of many of these signaling events has revealed that, in most cases, the mechanisms that mediate them have been conserved throughout metazoan evolution. Thus, the analysis of signaling pathways in C. elegans can aid in the understanding of signal transduction mechanisms in general. In this review we focus on signaling events that occur during the development of the hermaphrodite egg-laying system. Many of these signaling events occur at approximately the same time and in very close proximity to one another. Following a brief review of the individual signaling systems employed, we analyze the data that have started to address how the specificity among these pathways is maintained and how multiple pathways that affect individual developmental decisions are integrated. These issues are common to all signaling systems and should be instructive in presenting the complexities that are involved in obtaining a global understanding of development.

Effectiveness of injectable filler materials for smoothing wrinkle lines and depressed scars.

Both synthetic and biological materials have been used for smoothing wrinkle lines and depressed scars. Injectable fillers provide a convenient and easy alternative to surgical methods to treat dermal defects. Optimally, injectable fillers augmenting dermal tissue should exhibit properties similar to the dermal collagen network, providing a stable matrix for long-term correction but also being subject to the dynamic changes that occur to dermal tissue during the aging processes. At present, only biological materials are available on a commercial basis, although a number of synthetic injectables are being evaluated in clinical trials. The long-term effectiveness of most commercial materials has been limited prompting questions about the cost-benefits of such treatment. In this regard, the effectiveness of the injectable fillers, particularly of biological materials, will be discussed as will the difficulties involved in determining the expected result (effectiveness) for specific materials and in providing methods of measuring effectiveness.

Transport of glucose polymer-derived glucose by rabbit jejunum.

Mechanisms for the assimilation of glucose polymers have been inferred from perfusion studies. To further define these mechanisms, the results of measurements of unidirectional glucose fluxes across short-circuited rabbit jejunal segments in vitro are reported. Glucose polymer-stimulated short-circuit current was similar to that of glucose [19 +/- 6.0 microA/cm2 (n = 7) and 26 +/- 5.7 microA/cm2 (n = 13), respectively] and was inhibited by both acarbose and phlorizin. Acarbose, an alpha-glucosidase inhibitor with no effects of glucose transport, was used to uncouple digestion from absorption. Mucosal-to-serosal flux of glucose polymer-derived glucose was lower than that of an equal weight/volume of glucose [124 +/- 62 nmol.h-1.cm-2 (n = 4) vs. 452 +/- 121 nmol.h-1.cm-2 (n = 6); P less than 0.05] and was inhibited by both phlorizin and acarbose. No glucose polymers were detected in the serosal bath solutions by thin-layer chromatography. It is concluded that glucose polymer-derived glucose is transported by a phlorizin-inhibitable process at a rate slower than that of free glucose, a finding that suggests that hydrolysis limits glucose polymer assimilation.

Glucose flux from dietary disaccharides: all sugars are not absorbed at equal rates.

Considerable discrepancies exist in the literature regarding the rates of glucose absorption from the common dietary disaccharides, lactose, maltose, and sucrose. This study compared the unidirectional flux of glucose derived from dietary disaccharides with that of their constituent monosaccharides in vitro. Lactose-stimulated short-circuit current (Isc) and mucosal-to-serosal flux (Jm----s) were lower than that of an equimolar glucose-galactose mixture and were phlorizin inhibitable. Maltose- and glucose-stimulated Isc were similar, but Jm----s of glucose derived from the hydrolysis of maltose was lower than that of free glucose. Sucrose-stimulated Isc and Jm----s were similar to that of an equimolar glucose-fructose mixture. Isc and Jm----s of glucose from both maltose and sucrose were phlorizin and acarbose inhibitable. We conclude that the rate of glucose uptake from disaccharides is less than or equal to that of free glucose and is dependent on the glucose source. We speculate that regulation of glucose uptake from disaccharides can occur at three sites: the hydrolytic enzyme, the glucose transporter, and the tight junctions.

Long-term compatibility of intraocular lens implant materials.

This paper provides a general review of the long-term compatibility of intraocular lenses (IOLs), with specific reference to the compatibility of synthetic polymers used to fabricate IOLs. The first IOL was implanted in 1949. It was composed of polymethylmethacrylate (PMMA) and placed in the posterior chamber of the eye. In 1990, approximately 1.4 million IOLs were implanted in the U.S., nearly all composed of PMMA and nearly all implanted in the posterior chamber of the eye. However, many changes have taken place in the past 40 years to improve the compatibility of IOLs and to make cataract surgery and IOL implantation among the safest and most effective major surgical procedures.