Dynamic defect correlations dominate activated electronic transport in SrTiO3.

Strontium titanate (SrTiO3, STO) is a critically important material for the study of emergent electronic phases in complex oxides, as well as for the development of applications based on their heterostructures. Despite the large body of knowledge on STO, there are still many uncertainties regarding the role of defects in the properties of STO, including their influence on ferroelectricity in bulk STO and ferromagnetism in STO-based heterostructures. We present a detailed analysis of the decay of persistent photoconductivity in STO single crystals with defect concentrations that are relatively low but significantly affect their electronic properties. The results show that photo-activated electron transport cannot be described by a superposition of the properties due to independent point defects as current models suggest but is, instead, governed by defect complexes that interact through dynamic correlations. These results emphasize the importance of defect correlations for activated electronic transport properties of semiconducting and insulating perovskite oxides.

Free tensor fasciae latae flap for abdominal wall reconstruction: overview and new innovation.

Extensive abdominal wall defects may result from tumor extirpation, traumatic injury, or soft tissue infections. Extensive traumatic injuries can often disrupt the soft tissue content of the abdomen as well as the bony support provided by the pelvis. Reconstruction of the lower abdomen should aim to recreate dynamic stability. Five patients with extensive lower abdominal wall disruption following traumatic injuries or infection were treated using a novel flap for functional reconstruction. We devised a free neurotized osteomyocutaneous tensor fasciae latae (TFL) flap that would restore bony continuity by providing a vascularized bone graft and simultaneously maintain the integrity of the attachment of the tensor fascia latae muscle to the iliac crest, reestablishing musculofascial continuity. A branch of the superior gluteal nerve was harvested with this composite flap and coapted to an intercostal nerve for reinnervation, thereby creating a dynamic muscle in these patients. All patients underwent successful free tissue reconstruction with 100% flap survival. The lower abdominal wall and bony integrity of the pelvis were successfully reconstructed. Reinnervation has shown clinical signs of maintained dynamic stability. The innervated TFL osteomyocutaneous flap is an ideal option for lower abdominal reconstruction in patients with complex abdominoperineal defects with loss of bony integrity.

A novel model to measure the regenerative potential of the peripheral nervous system after experimental immunological demyelination.

BACKGROUND: Immunological demyelination is a proposed strategy to improve nerve regeneration in the peripheral nervous system. To investigate the remyelinating potential of Schwann cells in vivo in the peripheral nervous system, the authors have reproduced and expanded upon a novel model of immunological demyelination in the adult rat sciatic nerve. The authors demonstrate (1) the peripheral nervous system's quantitative, regenerative response to immunological demyelination and (2) whether Schwann cells within a region of demyelination are induced to divide in the presence of demyelinated axons. METHODS: The sciatic nerves of female Sprague-Dawley rats were exposed and injected with demyelinating agent bilaterally. At 3 days (n = 3), 7 days (n = 3), and 14 days (n = 3), the animals were euthanized for histological evaluation. A second group of animals (n = 3) was similarly injected with demyelinating agent and then exposed to bromodeoxyuridine between 48 and 72 hours after the onset of demyelination. These animals were euthanized soon after the last injection of bromodeoxyuridine. The tissue was analyzed for Schwann cells (labeled with antibodies to S100) and bromodeoxyuridine assay. RESULTS: A single epineural injection of complement proteins plus antibodies to galactocerebroside resulted in demyelination followed by Schwann cell remyelination. At 3 days after injection, peripheral nerve demyelination and Schwann cell proliferation were evident. Maximum demyelination was seen at 7 days; however, Schwann cell proliferation and remyelination peaked at 14 days after injection. CONCLUSIONS: These studies demonstrate an immunological model of demyelination and remyelination in the peripheral nervous system and quantitatively measure regenerative potential. This model will be used to isolate nerve segments and to measure their regenerative potential when given demyelinating agent after acute contusion and transection injuries.

Herpes simplex virus-thymidine kinase-based suicide gene therapy as a "molecular switch off" for nerve growth factor production in vitro.

Tissue-engineered constructs offer a new hope to patients suffering from functional impairment after nerve injury. An effort has been made to focus on delivery, regulation, and "molecular shutoff" of nerve growth factor (NGF) in tissue-engineered constructs. We have previously demonstrated that human embryonic kidney (HEK-293) cells can be genetically modified to secrete NGF at varying time points upon up regulation with Ponasterone A (PonA) both in vitro and in vivo. In the present study, HEK-293 cells that stably and inducibly produce NGF were further stably transfected with herpes simplex virus-thymidine kinase gene as a suicide gene (hNGF-EcR-293-TK) in order to shut off the NGF secretion and kill the cells upon treatment with ganciclovir (GCV). These cells following induction with PonA secreted NGF levels of 6659.2 +/- 489.4 pg/mL at day 10 postbooster dose at day 5, which was significantly higher than the control noninduced cells. The NGF secreted by these cells was bioactive as determined by a rat adrenal pheochromocytoma (PC-12) cell bioassay. Treatment of these cells with GCV significantly reduced the NGF levels to 645.3 +/- 16.2 pg/mL at day 10 and live cell numbers dropped to 7.95 x 10(3) +/- 278 compared to 2.73 x 10(5) +/- 6.1 x 10(4). GCV-treated cell media when transferred to the PC-12 cell bioassay demonstrated less than 10% cells differentiating into neurite-like extensions. We conclude that hNGF-EcR-293-TK cells can inducibly secrete bioactive NGF when treated with the inducing agent and can also be killed upon treatment with GCV. This double-gene transfection for gene expression and molecular shutoff mechanism will be a useful tool in tissue-engineered nerve constructs.

Transfer of training in robotic-assisted microvascular surgery.

BACKGROUND: Transfer of training refers to the ability to transfer acquired skills from one discipline to another. This study aims to determine whether experience in traditional freehand microsurgery facilitates mastery of robotic microsurgery. METHODS: Microsurgical anastomoses of coronary arteries harvested from explanted pig models were used to demonstrate whether prior experience with microsurgery is required in learning robot-assisted microsuturing. Eighty microsurgical anastomoses were performed. Three fully trained vascular surgeons (n = 3) (Group A) and 5 midlevel surgical residents (n = 5) (Group B) performed the anastomoses. Each subject performed 5 freehand and 5 robotic-assisted (Zeus robotic system) anastomoses. Anastomosis time and integrity of anastomoses were recorded, including errors of management (EOM) (breaking suture, breaking knots, breaking or damaging needles). RESULTS: For fully trained surgeons, all anastomoses in the robotic-assisted group were mechanically intact. There was significantly increased anastomosis time with the robot (Robot: 14 minutes, versus freehand: 7.2 minutes, P < 0.01). The robotic-assisted anastomoses were associated with a higher EOM (Robot: 1.2, versus freehand: 0.3, P < 0.01). Surgical trainees had longer anastomosis times with robotic assistance (Robot: 14.8 minutes, versus freehand, 12.7 minutes; P < 0.01) and increased EOM (Robot: 1.6, versus freehand: 1.0; P < 0.05).Overall, surgical trainees and fully trained vascular surgeons had longer anastomotic times with robotic assistance [Robot: 14.0 versus 14.8 minutes; P = not significant (NS)], and EOM (Robot: 1.6, versus freehand: 1.2; P = NS) were not significantly different. CONCLUSION: The technical feasibility of performing a safe and efficient robotic-assisted microsurgical anastomosis in explanted vessels was repeatedly tested and demonstrated in this study within reasonable time required for the anastomosis. Compared with conventional microanastomosis, both fully trained surgeons and residents demonstrated an ability to master the robotically assisted procedure with similarly longer anastomosis times and EOM. This study indicates that robotically assisted microanastomosis can be mastered equally well by surgical trainees and fully trained vascular surgeons.

Survival of diced cartilage grafts: an experimental study.

BACKGROUND: Use of diced cartilage grafts in rhinoplasty surgery has recently undergone a dramatic resurgence. Some authors recommend wrapping diced cartilage with oxidized methylcellulose (i.e., Surgicel). Others prefer wrapping diced cartilage with autogenous deep temporal fascia. This study was designed to compare the behavior of diced cartilage grafts as an isolated entity or when wrapped with either Surgicel or deep temporal fascia. METHODS: Septal cartilage and deep temporal fascia were obtained from five patients in this institutional review board-approved study. The cartilage was diced into 0.5-mm pieces and implanted into subcutaneous dorsal skin pockets of Rowlett nude rats as isolated diced cartilage grafts, or wrapped in Surgicel or deep temporal fascia. Pieces of Surgicel and fascia were implanted alone as controls. The specimens were harvested at 8 weeks; processed by thin-section histology; stained with hematoxylin and eosin, Masson's trichrome, glial fibrillary acidic protein, safranin-O, and Evans van Gieson; and evaluated to determine cartilage viability and architectural characteristics. RESULTS: Diced cartilage wrapped in Surgicel yielded the lowest percentage viability and minimal staining with hematoxylin and eosin, trichrome tissue, safranin-O, and Evans van Gieson stains. Diced cartilage grafts wrapped in fascia had the greatest percentage of viable cartilage. The grafts wrapped in deep temporal fascia also demonstrated the strongest staining with the aforementioned stains. Differences in uptake of glial fibrillary acidic protein were not noticeable between the three groups. However, absolute numbers of nucleated lacunae and basophilic lacunae were significantly higher for grafts wrapped in deep temporal fascia. CONCLUSIONS: Diced cartilage grafts have reemerged as a viable method for nasal reconstruction in both primary and secondary rhinoplasty. Wrapping diced cartilage specimens contains the individual pieces and facilitates graft placement. Surgicel wraps appear to incite an inflammatory response and subsequent absorption of the cartilage. Fascia wraps appear to minimize inflammatory responses to the cartilage, thereby preserving healthy cartilage. This study demonstrates that deep temporal fascia is the preferred envelope with which to facilitate graft containment and maintain chondrocyte viability of diced cartilage grafts.

Nerve growth factor expression response to induction agent booster dosing in transfected human embryonic kidney cells.

To assess whether nerve growth factor (NGF) expression would respond to booster dosing with the inducing agent ponasterone A, human embryonic kidney cells (HEK-293) were transfected with human NGF cDNA. Cells were cultured for 5 days in media with or without ponasterone A. On day 5, controls received a ponasterone A media replacement, whereas experimental groups received ponasterone A booster media replacement. NGF protein expression bioactivity was assessed using a PC-12 cell bioassay and the concentration of secreted NGF was quantified using NGF enzyme-linked immunosorbent assay. Cells with and without ponasterone A were left for 5 days without changing the medium. On day 5, the supernatants were collected and flash-frozen for enzyme-linked immunosorbent assay. The ponasterone A-positive and -negative booster medium was replaced in the appropriate wells. Supernatants were collected from the wells at 2, 4, and 6 days after the booster dose and removal of original supernatant. The medium was flash-frozen for enzyme-linked immunosorbent assay (1.5 ml), and the remaining 500 mul was transferred to PC-12 cells seeded onto 12-well plates to determine NGF bioactivity. All experiments were performed in quadruplicate. NGF production was measured daily by enzyme-linked immunosorbent assay over a 6-day period after the ponasterone A booster to a maximal release of 1233 +/- 130 pg/ml at day 6 (11 days after original induction). Maximal NGF production per 10(3) cells was 2.5 +/- 0.61 pg at day 6. Bioactivity was determined by percentage differentiation (per 100 cells counted) at 26, 52, and 98 percent for ponasterone A-treated wells on 2, 4, and 6 days after booster dosing (7, 9, and 11 days after induction), respectively. PC-12 cell differentiation was not visualized in the ponasterone A-negative control wells. Human NGF-EcR-293 cells can inducibly secrete bioactive NGF when exposed to the induction agent ponasterone A. Furthermore, repeated bioactive NGF expression peaks beyond that previously demonstrated can be achieved using induction agent booster dosing, indicating the ability to regulate the system over an extended period.

In vivo induction and delivery of nerve growth factor, using HEK-293 cells.

Tissue-engineering strategies offer hope to patients facing functional impairment after nerve injury. We have previously demonstrated that HEK-293 cells can release nerve growth factor (NGF) in vitro, using an inducible system of expression. In this study, our objective was to assess the efficacy of the NGF delivery system in vivo, using nude rats. HEK-293 cells were transfected with human NGF cDNA. Ponasterone A (PonA) was used as the inducing agent. NGF collection chambers were implanted subcutaneously in nude rats. Sealed chambers were filled with one of the following: (1) DMEM, (2) untransfected 293 cells (EcR-293) plus PonA, (3) untransfected EcR-293 without PonA, (4) transfected 293 cells (hNGF-EcR-293) plus PonA, or (5) transfected hNGF-EcR-293 without PonA. Chambers were aspirated 24, 48, and 120 h postimplantation. NGF secretion was analyzed in the following ways: (1) NGF protein expression bioactivity was assessed in a PC-12 cell bioassay, and (2) the concentration of secreted NGF was quantified by NGF ELISA. NGF quantification by ELISA reached a maximal release of 12.9 +/- 3.57 ng/mL at 120 h. PC-12 cells exposed to media from induced transfected HEK-293 cell chambers demonstrated higher levels of differentiation compared with controls. We conclude that hNGF-EcR-293 cells can inducibly secrete bioactive NGF when exposed to the induction agent PonA. This regulated delivery system can secrete bioactive NGF for up to 5 days in vivo. We believe this regulated delivery system will be useful for tissue-engineered nerve constructs.