Barcelona consensus on supermicrosurgery.

The popularity of supermicrosurgery has increased dramatically over the past few years, but the lack of agreement regarding the name of the technique and its applications has caused misunderstandings among microsurgeons when trying to communicate and compare surgical procedures. We report the consensus reached on the name used to refer to supermicrosurgery techniques following the First European Conference on Supramicrosurgery held in Barcelona (Spain) on March 4-5, 2010. Present applications, advantages, and disadvantages of supermicrosurgery are discussed. It was agreed that supermicrosurgery was the most accurate name to reflect the essence of this extremely delicate technique. According to Koshima, supermicrosurgery is a technique of microneurovascular anastomosis for vessels of 0.3 to 0.8 mm and single nerve fascicles. The range of applications for this technique has increased rapidly and now includes lymphedema treatment, nerve reconstruction, replantation and reconstruction of amputated fingertips, microsurgical flap salvage, and new possibilities for free tissue transfer. Supermicrosurgery is a remarkably useful reconstructive tool that involves a great deal of skill and has a steep learning curve for the microsurgeon to master. Although it is currently performed by only a minority of microsurgeons, we consider it will be incorporated into conventional microsurgery in the near future.

Designing Calcium Silicate Cements with On-Demand Properties for Precision Endodontics.

Calcium silicate (C3S) cements are available in kits that do not account for patients' specific needs or clinicians' preferences regarding setting time, radiopacity, mechanical, and handling properties. Moreover, slight variations in powder components and liquid content affect cement's properties and bioactivity. Unfortunately, it is virtually impossible to optimize several cement properties simultaneously via the traditional "one variable at a time" strategy, as inputs often induce trade-offs in properties (e.g., a higher water-to-powder ratio [W/P] increases flowability but decreases mechanical properties). Herein, we used Taguchi's methods and genetic algorithms (GAs) to simultaneously analyze the effect of multiple inputs (e.g., powder composition, radiopacifier concentration, and W/P) on setting time, pH, flowability, diametral tensile strength, and radiopacity, as well as prescribe recipes to produce cements with predicted properties. The properties of cements designed with GAs were experimentally tested, and the results matched the predictions. Finally, we show that the cements increased the genetic expression of odonto/osteogenic genes, alkaline phosphatase activity, and mineralization potential of dental pulp stem cells. Hence, GAs can produce cements with tailor-made properties and differentiation potential for personalized endodontic treatment.

Importance of preserving redundant pulp skin in thumb terminalization to allow subsequent reconstruction with on-top toe transfer: A clinical case (with video).

Traumatic thumb amputations can seriously jeopardise hand function. Several alternatives for reconstructing an opposable thumb have been described, including distraction osteogenesis, index finger pollicisation and free toe transfer. We present in this article the case of a young male patient that sustained a circular saw injury to his non-dominant thumb which resulted in loss of his distal phalanx. Preserving the sensate pulp allowed later reconstruction with an on-top free second toe transfer achieving a satisfactory outcome.

An early event in the interferon-induced transmembrane signaling process.

Human interferon stimulates a transient two- to threefold increase in the concentration of diacylglycerol and inositol tris-phosphate within 15 to 30 seconds of cell exposure to interferon. Antibodies to interferon inhibit this effect. The stimulation was measurable in isolated cell membranes exposed to interferon. Human alpha and beta, but not gamma, interferon stimulate this increase in cells containing the appropriate interferon receptor. The effect was proportional to the number of interferon receptors. Both the diacylglycerol increase and antiviral effects induced by interferon could be correlated in terms of dose dependence. Thus, a transient diacylglycerol increase is an early event in the interferon-induced transmembrane signaling process.

Use of magnetic resonance imaging in evaluation of placental invasion.

AIM: To review and describe the magnetic resonance imaging (MRI) features in patients with suspected placental invasion and correlate the findings with surgery and pathology findings. MATERIALS AND METHODS: A retrospective review was undertaken of the MRI images of seven consecutive patients with ultrasound findings suspicious for placental invasion. Two experienced MRI radiologists, blinded to the pathology and surgery findings, reviewed the MRI. The pathology or surgical findings were used as the reference standard to establish accuracy and concordance with the MRI findings. RESULTS: Three MRI features described in an earlier series were consistently present in the patients with placental invasion: lower uterine bulging, heterogeneous placenta, and dark intraplacental linear bands on T2-weighted images. CONCLUSION: MRI features, which were described in patients with placental invasion in an earlier series, were useful in establishing the presence and depth of placental invasion.

Assessing the reliability of the modified Gartland classification system for extension-type supracondylar humerus fractures.

PURPOSE: The Gartland extension-type supracondylar humerus (SCH) fracture is the most common paediatric elbow fracture. Treatment options range from nonoperative treatment (taping or casting) to operative treatments (closed reduction and percutaneous pinning or open reduction). Classification variability between surgeons is a potential contributing factor to existing controversy over treatment options for type II SCH fractures. This study investigated levels of agreement in extension-type SCH fracture classification using the modified Gartland classification system. METHODS: A retrospective review was conducted on 60 patients aged between two and 12 years who had sustained an extension-type SCH fracture and received operative or nonoperative treatment at a tertiary children's hospital. Baseline radiographs were provided, and surgeons were asked to classify the fractures as type I, IIA, IIB or III according to the modified Gartland classification. Respondents were then asked to complete a second round of classifications using reshuffled radiographs. Weighted kappa values were calculated to assess interobserver and intraobserver levels of agreement. RESULTS: In all, 21 paediatric orthopaedic surgeons responded to the survey and 15 completed a second round of ratings. Interobserver agreement for classification based on the Gartland criteria between surgeons was substantial with a kappa of 0.679 (95% confidence interval (CI) 0.501 to 0.873). Intraobserver agreement was substantial with a kappa of 0.796, (95% CI 0.628 to 0.864). CONCLUSION: Radiographic classification of extension-type SCH fractures demonstrated substantial agreement both between and within surgeon raters. Therefore, classification variability may not be a major contributing factor to the treatment controversy for type II SCH fractures and treatment variability may be due to differences in surgeon preferences. LEVEL OF EVIDENCE: III.

Energy Harvesting Combat Boot for Satellite Positioning.

Most portable electronic devices are power-limited by battery capacity, and recharging these batteries often interrupts the user's experience with the device. The product presented in this paper provides an alternative to powering portables by converting regular human walking motion to electricity. The device harvests electric power using air bulbs, distributed in the sole of a shoe to drive a series of micro-turbines connected to small DC motors. The number and position of air bulbs is optimized to harvest the maximum airflow from each foot-strike. The system is designed to continuously drive the micro-turbines by utilizing both outflow and inflow from the air bulbs. A prototype combat boot was fitted on the right foot of a 75 kg test subject, and produced an average continuous power on the order of 10 s of mW over a 22 Ω load during walking at 3.0 mph. This combat boot provides enough electric power to a passive GPS tracker that periodically relays geographical coordinates to a smartphone via satellite without battery replacement.

Characterisation of a high affinity Ca2+-stimulated, Mg2+-dependent ATPase in the rat parotid plasma membrane.

Two Ca2+-stimulated ATPase activities have been identified in the plasma membrane of rat parotid: (a) a (Ca2+ + Mg2+)-ATPase with high affinity for free Ca2+ (apparent Km = 208 nM, Vmax = 188 nmol/min per mg) and requiring micromolar concentration of Mg2+ and (b) a (Ca2+ or Mg2+)-ATPase with relatively low affinity for free Ca2+ (K0.5 = 23 microM) or free Mg2+ (K0.5 = 26 microM). The low-affinity (Ca2+ or Mg2+)-ATPase can be maximally stimulated by Ca2+ alone or Mg2+ alone. The high-affinity (Ca2+ + Mg2+)-ATPase exhibits sigmoidal kinetics with respect to ATP concentration with K0.5 = 0.4 mM and a Hill coefficient of 1.91. It displays low substrate specificity with respect to nucleotide triphosphates. Although trifluoperazine inhibits the activity of the high affinity (Ca2+ + Mg2+)-ATPase only slightly, it inhibits the activity of the low-affinity (Ca2+ or Mg2+)-ATPase quite potently with 22 microM trifluoperazine inhibiting the enzymic activity by 50%. Vanadate, inositol 1,4,5-trisphosphate, phosphatidylinositol 4,5-bisphosphate, Na+,K+ and ouabain had no effect on the activities of both ATPases. Calmodulin added to the plasma membranes does not stimulate the activities of both ATPases. The properties of the high-affinity (Ca2+ + Mg2+)-ATPase are distinctly different from those of the previously reported Ca2+-pump activity of the rat parotid plasma membrane.

Catalytic and regulatory properties of two forms of bovine heart cyclic nucleotide phosphodiesterase.

The soluble supernatant fraction of bovine heart homogenates may be fractionated on a DEAE cellulose column into two cyclic nucleotide phosphodiesterases (EC 3.1.4.-):PI and PII phosphodiesterases, in the order of emergence from the column. In the presence of free Ca2+, the PI enzyme may be activated several fold by the protein activator which was discovered by Cheung((1971) J. Biol. Chem. 246, 2859-2869). The PII enzyme is refractory to this activator, and is not inhibited by the Ca2+ chelating agent, ethylene glycol bis (beta-aminoethyl ether)-N, N'-tetraacetate (EGTA). The activated activity of PI phosphodiesterase may be further stimulated by imidazole or NH+4, and inhibited by high concentrations of Mg2+. These reagents have no significant effect on either the PII enzyme or the basal activity of PI phosphodiesterase. Although both forms of phosphodiesterase can hydrolyze either cyclic AMP or cyclic GMP, they exhibit different relative affinities towards these two cyclic nucleotides. The PI enzyme appears to have much higher affinities toward cyclic GMP than cyclic AMP. Km values for cyclic AMP and cyclic GMP are respectively 1.7 and 0.33 mM for the non-activated PI phosphodiesterase; and 0.2 and 0.007 mM for the activated enzyme. Each cyclic nucleotide acts as a competitive inhibitor for the other with Ki values similar to the respective Km values. In contrast with PI phosphodiesterase, PII phosphodiesterase exhibits similar affinity toward cyclic AMP and cyclic GMP. The apparent Km values of cyclic AMP and cyclic GMP for the PII enzyme are approx. 0.05 and 0.03 mM, respectively. The kinetic plot with respect to cyclic GMP shows positive cooperativity. Each cyclic nucleotide acts as a non-competitive inhibitor for the other nucleotide. These kinetic properties of PI and PII phosphodiesterase of bovine heart are very similar to those of rat liver cyclic GMP and high Km cyclic AMP phosphodiesterases, respectively (Russel, Terasaki and Appleman, (1973) J. Biol. Chem. 248, 1334).

Cloning, characterization and expression of a cDNA clone encoding rabbit ubiquitin-conjugating enzyme, E2(32k).

A cDNA clone encoding rabbit E2(32k) was obtained by library screening and PCR. The cDNA contains an open reading frame coding for 238 amino acids which shows an overall identity of 81% to human CDC34, the cell cycle-related ubiquitin-conjugating enzyme. A 50% homology to yeast CDC34 within the conserved core domain was also observed. Northern blot analysis indicated that three transcripts existed in all six rabbit tissues examined but their expression levels varied over a wide range. The putative cDNA coding region was highly expressed in Escherichia coli as a his-tagged protein which was purified to homogeneity. The ability of this expressed protein to form a thiolester bond with ubiquitin showed that it was functionally active. The ability of this protein to catalyze the conjugation of ubiquitin to histone H2A and H2B was also examined.

Feasibility of a novel blood noise reduction algorithm to enhance reproducibility of ultra-high-frequency intravascular ultrasound images.

BACKGROUND: Ultra-high-frequency (40- to 50-MHz) intravascular ultrasound (IVUS) improves image quality compared with conventional 20- to 30-MHz IVUS. However, as the frequency of IVUS increases, high-intensity backscatter from blood components may cause visual difficulties in discrimination between the lumen and arterial wall structure. The purpose of this study was to evaluate the effect of a novel blood noise reduction algorithm (BNR) on quantitative coronary ultrasound measurements. METHODS AND RESULTS: IVUS studies using a 40-MHz transducer were performed in 35 patients with coronary artery disease. A total of 620 gray-scale images (310 pairs) were processed with and without the BNR, and lumen cross-sectional area (CSA) was determined by 2 independent observers. With the BNR, the intraobserver and interobserver correlation coefficients for lumen CSA were significantly improved (0.85 to 0.99 and 0.80 to 0.98, respectively). In the 270 images (135 pairs) in which vessel wall measurements were possible, the BNR significantly improved the intraobserver and interobserver correlation coefficients for plaque plus media CSA (0.83 to 0.99 and 0.76 to 0.97, respectively), whereas no influence was observed for external elastic membrane CSA (1.00 to 1.00 and 0.99 to 0.99, respectively). CONCLUSIONS: This study demonstrates the feasibility of this novel algorithm to reduce blood noise, thereby enabling accurate lumen border delineation and providing reproducible measurements of both the lumen and plaque plus media CSAs. Incorporating a digital BNR may serve as an important adjunct to ultra-high-frequency IVUS imaging for improving accurate quantitative evaluation of vessel dimensions.

Preoperative management of the amputated limb.

Little literature exists on storage and preparation of an amputated limb prior to transfer to a reconstructive plastic surgical unit for possible macroreplantation. This paper describes practical measures used to prolong ischaemia time allowing macroreplantation, tissue harvesting, or fashioning of a useful stump. A simple protocol is used to summarise these points.

Rabbit ubiquitin-activating enzyme E1: cDNA cloning, sequence and expression.

A cDNA clone encoding ubiquitin-activating enzyme E1 has been isolated from a rabbit heart cDNA library and sequenced. The 3.485 kb cDNA contains an open reading frame of 1058 amino acid residues which predicts a protein of approx. 118 kDa. The deduced protein sequence exhibits a very high homology to other ubiquitin-activating enzymes identified in a variety of organisms. Northern blot analysis reveals a single transcript of approx. 3.5 kb in all the rabbit tissues examined. The entire coding region of the rabbit E1 cDNA has been expressed as a his-tagged protein. The recombinant protein has been verified by its ability to cross-react with anti-human E1 antibodies. Ubiquitin thiolester assay shows that the recombinant rabbit E1 protein is functional.

Cloning and characterization of AWP1, a novel protein that associates with serine/threonine kinase PRK1 in vivo.

We describe the cloning and expression of cDNAs encoding a novel human protein of 208 amino acid residues with a predicted molecular mass of 22.6kDa and its mouse homologue. We name this protein as AWP1 (associated with PRK1). AWP1 is a ubiquitously expressed protein, and the Awp1 gene is switched on during early human and mouse development. When expressed in COS-1 cells, the Myc-tagged AWP1 has an apparent molecular mass higher than that deduced from its amino acid sequence. AWP1 possesses a conserved zf-A20 zinc finger domain at its N-terminal and a zf-AN1 zinc finger domain at its C-terminal. Co-immunoprecipitation experiments revealed that mouse AWP1 specifically interacts with a rat serine/threonine protein kinase PRK1 in vivo. Hence, AWP1 may play a regulatory role in mammalian signal transduction pathways.

Identification of two proteins, S14 and UIP1, that interact with UCH37.

By the use of the yeast two-hybrid screen we have identified two proteins that interacted with UCH37: S14, which is a subunit of PA700 and a novel protein, UIP1 (UCH37 interacting protein 1). The interaction of UCH37 with S14 or UIP1 was confirmed by in vitro binding assay and in vivo co-immunoprecipitation analysis. The C-terminal extension of UCH37 is essential for interaction with S14 or UIP1 as shown by the yeast two-hybrid assay and the in vitro binding assay. Furthermore, UIP1 blocked the interaction between UCH37 and S14 in vitro.

Short-term TPN containing n-3 fatty acids ameliorate lactic acidosis induced by endotoxin in guinea pigs.

We evaluated the effect of total parenteral nutrition (TPN) enriched with n-3 fatty acids on the physiologic response to endotoxin in guinea pigs. Animals were randomly assigned to receive TPN differing only in lipid source for 3.5 d. Group 1 received soybean fat emulsion (Intralipid) whereas group 2 received fish (menhaden) oil. During the last 7 h of TPN, animals were further randomized to have either saline or E coli endotoxin added to the infusate. Acid-base status and serum lactate concentrations were determined. Animals infused with soybean fat emulsions and endotoxin developed a significant metabolic acidosis, lactic acidemia, and decrease in mixed venous O2 compared with controls and fish-oil-treated animals (p less than 0.05). The significantly reduced serum lactate and higher mixed venous O2 in fish-oil-infused animals suggests that the underlying mechanism involves improvement in endotoxin-induced tissue hypoperfusion, presumably through alterations in prostaglandin metabolism.

Effect of myricetin and other flavonoids on the liver plasma membrane Ca2+ pump. Kinetics and structure-function relationships.

Thirty-three different flavonoids were screened for their ability to influence ATP-dependent Ca2+ uptake by rat liver plasma membrane vesicles. Nine of the flavonoids, at a concentration of 100 microM inhibited Ca2+ uptake by more than 20%. The remaining 24 flavonoids exhibited little or no effect. The relative order of potency of the more biologically active flavonoids was myricetin greater than butein greater than phloretin = luteolin greater than eriodictyol = silybin. Myricitrin and phloridzin, the glycosides of myricetin and phloretin, respectively, had no effect. The degree of inhibition caused by myricetin was concentration dependent and was also affected by the preincubation time. After 10 min of preincubation, 52 microM myricetin lowered the initial rate of 45Ca uptake by 50%. The inhibition by myricetin was non-competitive with respect to Mg-ATP and of a mixed type with respect to Ca2+. At a concentration of 100 microM, myricetin had no effect on several plasma membrane enzymes such as 5'-nucleotidase, alkaline phosphatase and a Ca2(+)-activated ATPase but inhibited K(+)-dependent p-nitrophenyl phosphatase by 83%. The ATP-dependent Ca2+ transport systems located on the plasma membrane or endoplasmic reticulum derived from other tissues were also inhibited by myricetin. Analysis of the structure-activity relationship revealed that lipid solubility and polyhydroxylation particularly at positions 5,7,3' and 4' of the flavonoid ring structure enhanced the ability of the flavonoid to inhibit Ca2+ uptake. The results suggest that inhibition of Ca2+ transport activity probably involves the interaction of the phenolic groups of the flavonoid with the Ca2+ transporting protein.

Long-term feeding with structured lipid composed of medium-chain and N-3 fatty acids ameliorates endotoxic shock in guinea pigs.

The metabolic and physiologic responses to 7-hour endotoxin infusion (5.0 mg/kg h) were evaluated in guinea pigs following 6 weeks of dietary enrichment with diets containing either chemically structured lipid (SL) composed of medium-chain triglycerides (MCT) and long-chain triglycerides (LCT) in the form of N-3 polyunsaturated fatty acids (PUFAs), or safflower oil (SO), which is high in N-6 fatty acids. Plasma phospholipid fatty acid profiles, arterial blood pH, PCO2, PO2, HCO3, lactate, blood pressure, oxygen consumption, and energy expenditure were examined. Plasma phospholipid fatty acids profiles reflected dietary intake with SL-fed animals demonstrating a significantly higher N-3 to N-6 fatty acid ratio compared with SO-fed animals. SL-fed animals responded to endotoxemia with a mild metabolic acidosis with respiratory compensation, which was associated with moderate lactatemia (3 mmol/L). SO-fed animals developed a severe metabolic acidosis with acidemia and respiratory compensation, which was associated with hyperlactatemia (8 mmol/L, P less than .05 v SL). No differences were observed in blood pressure, oxygen consumption, energy expenditure, or respiratory quotient during endotoxemia between dietary groups compared with controls. We conclude that diets enriched with structured lipid composed of medium-chain and N-3 fatty acids can attenuate the sequelae of endotoxemia.