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The feasibility and safety of fluoroscopy-guided percutaneous intramyocardial gene injection in porcine heart.

BACKGROUND: Catheter-based transendocardial gene injection would be useful for the delivery of genes into the heart. We examined the feasibility and safety of percutaneous intramyocardial gene injections with fluoroscopic guidance alone. METHODS: We performed the procedure through an 8F arterial sheath inserted into the left carotid artery. In protocol 1, a mixture of India ink and normal saline was injected through a needle injection catheter in six pigs. We monitored blood pressure and ECG continuously during the procedure. Echocardiography, left ventriculography, and coronary angiography were performed. All pigs were sacrificed 2 days later and hearts were harvested. In protocol 2, a mixture of India ink and plasmid encoding CAT gene was injected in the same manner in eight pigs. Myocardial tissue was obtained 7 days after the procedure to assess gene expression. In protocol 3, four pigs were intentionally needle-perforated in the ventricular wall and were observed for 7 days. RESULTS: In protocol 1, there was no significant hemodynamic changes or serious arrhythmias during the procedure. Echocardiography and angiography revealed no evidence indicating pericardial effusion or wall motion abnormalities. Harvested hearts revealed one intramyocardial hematoma in a total of 36 injection sites. In protocol 2, the gene expression could be identified in 39 sites out of 48 injections after 7 days. In protocol 3, no animal showed signs indicating cardiac tamponade during the observation period. CONCLUSIONS: Our data suggest that fluoroscopy-guided percutaneous intramyocardial gene injection is a feasible and safe procedure, with no indication of associated significant hemodynamic changes, arrhythmias, or mortality.

Administration, Cutaneous↗

The prefabrication of a bone graft in a rat model.

The prefabrication of bone grafts in a rat model was investigated. In 26 Sprague-Dawley rats, free iliac crest bone graft was harvested, bivalved, and reinserted heterotopically into the groin, where it was closed around the mobilized superficial inferior epigastric vascular bundle. In half the animals, the vascular bundle remained in continuity as a flow-through pedicle (group 1); in the remaining animals, the pedicle was ligated and divided distal to the bone graft. All grafts were isolated from other tissues by a silicone sheet envelope. At 3 or 6 weeks, the grafts were re-explored and analyzed by India ink perfusion and histologic examination for evidence of viability and neovascularization. Three weeks after insertion, India ink perfusion of the group 1 and 2 grafts revealed neovascularization extending to the periphery of the graft, and histologic examination showed extensive new bone formation on endosteal, periosteal, and trabecular surfaces of the graft. Six weeks after insertion, creeping substitution had almost completely remodelled the cortical and cancellous bone of both group 1 and 2 grafts to create a viable vascularized bone graft on a pedicle. In 3 control nonvascularized grafts (free iliac cortical bone without an implanted pedicle), all pre-existing bone of the graft was dead 3 weeks after insertion, and only very limited new bone formation was present within the graft.

Animals↗

Cellular and molecular mechanisms of stress-induced premature senescence (SIPS) of human diploid fibroblasts and melanocytes.

Replicative senescence of human diploid fibroblasts (HDFs) or melanocytes is caused by the exhaustion of their proliferative potential. Stress-induced premature senescence (SIPS) occurs after many different sublethal stresses including H(2)O(2), hyperoxia, or tert-butylhydroperoxide. Cells in replicative senescence share common features with cells in SIPS: morphology, senescence-associated beta-galactosidase activity, cell cycle regulation, gene expression and telomere shortening. Telomere shortening is attributed to the accumulation of DNA single-strand breaks induced by oxidative damage. SIPS could be a mechanism of accumulation of senescent-like cells in vivo. Melanocytes exposed to sublethal doses of UVB undergo SIPS. Melanocytes from dark- and light- skinned populations display differences in their cell cycle regulation. Delayed SIPS occurs in melanocytes from light-skinned populations since a reduced association of p16(Ink-4a) with CDK4 and reduced phosphorylation of the retinoblastoma protein are observed. The role of reactive oxygen species in melanocyte SIPS is unclear. Both replicative senescence and SIPS are dependent on two major pathways. One is triggered by DNA damage, telomere damage and/or shortening and involves the activation of the p53 and p21(waf-1) proteins. The second pathway results in the accumulation of p16(Ink-4a) with the MAP kinase signalling pathway as possible intermediate. These data corroborate the thermodynamical theory of ageing, according to which the exposure of cells to sublethal stresses of various natures can trigger SIPS, with possible modulations of this process by bioenergetics.

Cell Cycle↗

Laser eradication of pigmented lesions and tattoos.

Laser treatment of pigmented lesions can be a rewarding experience when appropriate lesions are treated. Accurate diagnosis of pigmented lesions is needed before treatment. In some lesions, adjuvant topical therapy is greatly beneficial, and for others it may be the only option. The treatment of melanocytic nevi is controversial but worth pursuing. All tattoos respond well to Q-switched lasers; the appropriate wavelength depends on the color of ink. Amateur and traumatic tattoos clear readily with laser treatment. Cosmetic tattoos should be approached with caution. In addition, the use of laser-responsive ink and higher-powered or shorter-pulsed (picosecond) lasers may further enhance the ability to treat tattoos.

Follow-Up Studies↗

Prefrontal regions play a predominant role in imposing an attentional 'set': evidence from fMRI.

fMRI was used to determine whether prefrontal regions play a predominant role in imposing an attentional 'set' that drives selection of task-relevant information. While monitoring for an atypical item, individuals viewed Stroop stimuli that were either colored words or colored objects. Attentional demands were varied, being greater when the stimuli contained two distinct and incongruent sources of information about the task-relevant attribute (e.g., when attending to color, seeing the word 'blue' in red ink) as compared to only one source (e.g., seeing the word 'late' in red ink). Prefrontal but not anterior cingulate regions exhibited greater activation on incongruent than neutral trials, suggesting that prefrontal cortex has a major role in imposing an attentional 'set'. In addition, we found that prefrontal activation is most likely to occur when that attentional set is difficult to impose.

Attention↗

Development of disposable amperometric sulfur dioxide biosensors based on screen printed electrodes.

The possibility of developing amperometric biosensors for the measurement of SO(2) in flowing gas streams has been examined. Screen-printed carbon electrodes (SPCEs) were tailored with the enzyme sulfite oxidase and cytochrome c and the response is generated through the resulting enzymatic and electrocatalytic reactions involving SO(3)(2-), formed when SO(2) gas is dissolved in the supporting electrolyte. Two methods of integrating the enzyme and cytochrome c with the SPCE were investigated. In one design (b-type biosensor), the components were mixed thoroughly with the same ink used to produce the SPCEs, then the modified ink was spread over the working electrode. In the second approach the bio-components were dissolved in the supporting electrolyte and simply deposited on top of the transducer (s-type biosensor). Both devices gave linear responses over the range 4--50 ppm but the sensitivity of the s-type was approximately twice that of the b-type biosensor. In addition, the time taken to reach 90% of the maximum response (t(90%)) was 110 s for the s-type biosensor compared with 200 s for the b-type biosensor. These studies illustrate the successful use of biosensors for the detection of sulfur dioxide at the relatively low potential of +0.3 V versus Ag.AgCl and should provide useful alternatives for decentralised environmental studies.

Biosensing Techniques↗

Blood supply of small fibula segments: an experimental study on human cadavers.

When vascularized fibula grafts are used in mandibular reconstruction, it is often necessary to divide the graft into segments so as to follow the mandibular curvature. The periosteal blood supply to the segments is usually sufficient. It is unclear, however, how short the segments can be without compromising the blood supply. The present study sought to clarify this issue by injecting Indian ink into the afferent vessels in human cadavers. The evaluation of 300 specimens showed the ink perfusion to be significantly higher in segments longer than 1 cm than in shorter segments. Furthermore, the perfusion in the cortical outer third of the shaft was markedly higher than in the cortical inner two-thirds. The staining was significantly more intense in the younger (32-55 years) than in the older (56-73 years) age group. With correct dissection and successful anastomosis, however, fibula segments 1 cm or longer are probably sufficiently perfused, independent of the patient's age. The perfusion in shorter segments is uncertain. Clinically this means that segments shorter than 1 cm ought to be used only in a surgically non-compromised recipient site, where they could also take as non-vascularized grafts.

Adult↗

The extraneural and intraneural arterial anatomy of the ulnar nerve at the elbow.

The purpose of this report was to investigate the vascular anatomy of the ulnar nerve at the elbow with a technique of combined India ink and latex injection followed by nondissection chemical debridement. Twenty-two fresh human cadaveric arms were injected with India ink to stain the intraneural microcirculation followed immediately by latex injection and chemical debridement for study of the extraneural vascularization. After clearing with a modified Spalteholtz technique, the intraneural blood supply was studied in 10 of the specimens. The findings demonstrated a consistent but segmental extraneural and intraneural vascular supply from the superior ulnar collateral, inferior ulnar collateral, and posterior ulnar recurrent arteries. No identifiable direct anastomosis was seen between the superior ulnar collateral and posterior ulnar recurrent arteries in 20 of 22 arms. The inferior ulnar collateral artery provided the only direct vascularization to the nerve in the region just proximal to the cubital tunnel. Although the clinical importance of maintaining specific arterial sources to the ulnar nerve has not been determined, these anatomic findings indicate that the arterial contribution from the inferior ulnar collateral artery may be more important than appreciated previously.

Adult↗

Evaluation of phagocytic function of macrophages in rats after partial splenectomy.

BACKGROUND: Understanding the immunologic properties of the spleen has enabled surgeons to practice splenic conservation surgery. If the upper pole of the spleen can be preserved solely on the upper short gastric vessels, will phagocytic function of macrophages in remnant splenic tissue be affected? The aim of this experimental study was to evaluate the phagocytic function of macrophages in partially resected spleens, with hilar excision preserving the short gastric vessels. STUDY DESIGN: Forty-eight female Wistar albino rats were divided into four groups. Groups 1 and 2 underwent sham operations and groups 3 and 4 underwent partial splenectomy. One milliliter of sodium chloride 0.9% was injected into the abdomen of the rats in groups 1 and 3 and 1 mL of Streptococcus pneumoniae type III as an antigenic stimulus was injected into the abdomen of the rats in groups 2 and 4, 6 weeks after the first operation. Forty-eight hours later, relaparotomy was performed in all animals. India ink was used to determine the capacity of uptake in the splenic phagocytes. To evaluate the phagocytic function of the splenic tissues, histologic examinations were performed according to a macrophage grading system. RESULTS: All spleens in all four groups were stained black after injection of India ink. Phagocytic activity of macrophages was reduced in the partially splenectomized groups, compared with intact spleen groups (group 3 versus group 1; p < 0.0001, group 4 versus group 2; p < 0.0001). There was a significant difference between groups 1 and 2 according to phagocytic function of macrophages (p = 0.0121). Also, after Streptococcus pneumoniae type III injection as an antigenic stimulus in group 4, we found that the phagocytic functions of macrophages increased compared with those of the sodium chloride 0.9%-injected group 3 after partial splenectomy (p < 0.0001). CONCLUSIONS: Phagocytic function of macrophages in rats decreased after partial splenectomy. Nevertheless, the remnant spleens in rats could be stimulated when challenged with an antigenic stimulus.

Animals↗

Bacteroides fragilis adherence to Caco-2 cells.

The ability of ten Bacteroides fragilis strains isolated from intestinal and non-intestinal infections, normal flora and environment to adhere to human colon carcinoma cells, Caco-2, was examined. The adherence capacity varied among the strains tested from strongly adherent (76-100%) to non- or weakly adherent (0-25%). Negative staining with Indian ink showed that all the strains were capsulated, although strain 1032 (strongly adherent and originated from bacteremia) had the highest rate of capsulated cells in the culture. All strains studied presented an electron-dense layer and no fimbrial structures in their surface after PTA negative staining. The analysis of the strains with ruthenium red showed the presence of an acidic polysaccharide and also surface vesicles in all of them. The strain 1032 presented an aggregative adherence pattern toward Caco-2 cells monolayers. It could be seen trapped by elongated microvilli and surrounded by extracellular material in the scanning electron microscope. Treatment with sodium periodate (100 mM/1 h) reduced significantly its adherence capacity and also the expression of an electron-dense layer and of the capsule, detected with PTA and Indian ink staining, respectively. We suggest that the capsular polysaccharide might mediate the adherence of the B. fragilis to Caco-2 cells.

Journal Article↗

Development and evaluation of an inexpensive sensor system for use in measuring relative finger positions.

An inexpensive sensor was developed using a carbon-based electrically conductive ink. A correlation of the actual flex angle to the measured flex angle of the sensors was obtained by experimentally determining a third-order polynomial that represented the response of the sensor and its hardware system (r = 0.999). The response time, when going from an angle of 0-90 degrees, was extremely good. The sensors exhibited an acceptable dynamic response with an error of less than 5% when flexed repeatedly to an angle of 90 degrees. A second-order polynomial was found to express the resistance as a function of angle (r = 0.999) for all of the sensors tested. Although these sensors are not as precise as the more expensive sensors that are available, it was estimated that the production cost of these sensors was less then US $0.50 a piece. In addition, there are many ways that the sensor production method might be improved to produce more accurate sensors using carbon-based electrically conductive ink. While much additional work is still necessary, the system presented in this paper could be incorporated into a complete device to monitor the rehabilitation of an impaired hand. Unlike other systems that are currently available commercially, this system is inexpensive, easily manufactured, accurate, and could be readily used in a variety of clinical settings.

Biomechanical Phenomena↗

An evaluation of line quality in photocopied signatures.

Qualified forensic document examiners from Canada, USA, Great Britain and Australia participated in this study. They were sent one photocopied signature, together with a set of original known standards to evaluate the line quality of the photocopied signature and compare these characteristics with the standards. They were asked to assess the photocopies for the presence of tremor, hesitation, pen lifts, patching, blunt initial or terminal strokes, and abrupt changes in direction. The authors compared the assessments with the corresponding original signatures. It was possible to determine which line quality characteristics were the most difficult to detect, which indicia of simulation were concealed and what anomalies were introduced by the photocopying process. Results indicated that line quality characteristics can be identified in average quality photocopies. Furthermore, accurate judgements can be made regarding similarities and differences in line quality when comparing a photocopied signature with original known standards. In a few instances, ink line morphology was problematic when striation patterns and ink gooping were misinterpreted as pen lifts, hesitation and patching on the photocopies.

Copying Processes↗

Integrating polymerase chain reaction, valving, and electrophoresis in a plastic device for bacterial detection.

An integrated plastic microfluidic device was designed and fabricated for bacterial detection and identification. The device, made from poly(cyclic olefin) with integrated graphite ink electrodes and photopatterned gel domains, accomplishes DNA amplification, microfluidic valving, sample injection, on-column labeling, and separation. Polymerase chain reaction (PCR) is conducted in a channel reactor containing a volume as small as 29 nL; thermal cycling utilizes screen-printed graphite ink resistors. In situ gel polymerization was employed to form local microfluidic valves that minimize convective flow of the PCR mixture into other regions. After PCR, amplicons (products) are electrokinetically injected through the gel valve, followed by on-chip electrophoretic separation. An intercalating dye is admixed to label the amplicons; they are detected using laser-induced fluorescence. Two model bacteria, Escherichia coli O157 and Salmonella typhimurium, were chosen to demonstrate bacterial detection and identification based on amplification of several of their unique DNA sequences. The limit of detection is about six copies of target DNA.

Bacteria↗

Analysis of non-Newtonian liquids using a microfluidic capillary viscometer.

We have used a self-calibrating microfabricated capillary viscometer to analyze non-Newtonian power law fluids. The capillary viscometer can generate a wide range of shear rates during a single experimental run enabling quick and accurate analysis of non-Newtonian liquids. The measurement of viscosity is based on monitoring the capillary pressure-driven movement of fluid sample whose mean velocity and, therefore, shear rate varies with time. The device has been successfully tested for accuracy and robustness with dilute as well as semidilute solutions of flexible elastic polymers including poly(ethylene oxide) and hydrolyzed polyacrylamide to an aqueous solution of a stiff rodlike polymer molecule of xanthan gum, a popular emulsifier and food thickener, as well as with ink-jet printing inks. Viscosities in the range of 1-600 cP were measured, and shear rates varying from 5 to 1000 s(-1) have been obtained on the microfabricated viscometer with the current geometry and channel dimensions. The total measurement time varied between 2 and 8 min and less than 1 microL of sample volume was required. Such a microfabricated capillary viscometer would have possible applications in quality control and manufacturing where rapid and repeated measurements need to be made using limited sample volume.

Biopolymers↗

London University Search Instrument: a combinatorial robot for high-throughput methods in ceramic science.

This paper describes the design, construction, and operation of the London University Search Instrument (LUSI) which was recently commissioned to create and test combinatorial libraries of ceramic compositions. The instrument uses commercially available powders, milled as necessary to create thick-film libraries by ink-jet printing. Multicomponent mixtures are prepared by well plate reformatting of ceramic inks. The library tiles are robotically loaded into a flatbed furnace and, when fired, transferred to a 2-axis high-resolution measurement table fitted with a hot plate where measurements of, for example, optical or electrical properties can be made. Data are transferred to a dedicated high-performance computer. The possibilities for remote interrogation and search steering are discussed.

Aluminum Oxide↗

Multielemental analysis of purpleback flying squad using high resolution inductively coupled plasma-mass spectrometry (HR ICP-MS).

Forty-four elements were analyzed in 21 tissues of purpleback flying squid, Sthenoteuthis oualaniensis, by high resolution inductively coupled plasma-mass spectrometry (HR ICP-MS) and inductively coupled plasma atomic emission spectrophotometry (ICP-AES). Greater concentrations of V, Fe, Co, Ni, Cu, Ag, Cd, Pb, and Bi were found in liver, pancreas, and ink sac than in other tissues. Ink sac concentrated remarkable levels of Ca and Sr in addition to the above-mentioned elements. Several alkalis, alkaline earth, and rare earth elements preferentially accumulated in muscle. Among the hard tissues, accumulation of V and U in beak, Ni, Zn, and Cd in gladius and Cr in skin was prominent. K, Rb, Cs, Pb, Bi and some transition elements (V, Co, Cu, Zn, Ag, Cd) were significantly (p < 0.05) higher in the livers of adult than in juvenile squids. Sodium, alkaline earth, and rare earth elements were higher in the livers of juveniles than in adult squids.

Age Factors↗

Syntheses and structures of zwitterionic indium(III) and di-zinc compounds of an extended nitrogenous ligand. Examples of unusually long wavelength transitions in d10-metal complexes.

In acetonitrile, the reaction of hydrated InCl(3) and 2-[(2-(pyridylamino)phenylazo]pyridine [HL] affords an ink-blue octahedral indium(III) complex having the formula [InCl(3)(HL)]. The compound is a zwitterion in which the positive and the negative charges reside on the extended ligand HL. The secondary amine nitrogen in it is deprotonated, while the free pyridyl nitrogen is protonated. The reaction of ZnCl(2), on the other hand, produces a di-metallic complex, [Zn(2)Cl(2)L(2)], where the two metal ions are bridged across the deprotonated ligand [L](-). The color of the di-zinc complex is also ink blue. Interestingly, the UV-vis spectrum of the indium compound is almost identical to that of the di-zinc compound. Long-wavelength transitions near 590 nm in these examples are assigned to intraligand pi-pi transitions.

Journal Article↗

Miniaturized chemical multiplexed sensor array.

Miniaturized tin oxide semiconductor sensors are fabricated directly by site-specific dip-pen nanopatterning using precursor inks derived from the sol-gel method. The good flow characteristics and strong affinity of the sols to measurement electrodes enable intimate contact. The measurable, reproducible, and proportionate changes in the resistance of the sensors when exposed to trace quantities of oxidative and reducing gases constitute the basis for such sensors. These sensors show rapid response and ultrafast recovery for the detection of nitrogen dioxide and acetic acid. Furthermore, an array of eight miniaturized sensors is created by doping the pristine tin-based sol ink with different metal ions; the different responses of each sensor to certain gases constitute a reference response spectrum that can be used to recognize the gas. Such recognition ability, instant response and rapid recovery, compact size, and integration with the current microelectronics platform make the miniaturized sensor array a significant development for the on-site and real-time detection of life-threatening gases.

Journal Article↗