The influence of endocardial endothelium on myocardial contraction.

A novel unidentified agent, provisionally named 'endocardin', has been shown to be released from endocardial endothelium. Endocardin has a unique prolonging effect on myocardial contraction. In contrast, endothelium-derived relaxing factor released from endocardial endothelium has the opposite effect of abbreviating contraction. Jerry Smith and colleagues discuss the mechanisms of action of these agents and their possible physiology and pathophysiology.

Oligosaccharide-based block copolymers: metal-free thiol-maleimide click conjugation and self-assembly into nanoparticles.

Amphiphilic oligosaccharide-based block copolymers (OBCPs) are able to self-assemble either into nanoparticles with biocompatible oligosaccharides corona in aqueous solution or in sub-nanopatterned thin-films originating from the high incompatibility between the different blocks. For these reasons, these biosourced OBCPs are valuable structures for applications in nanomedicine and nanoelectronics. Up to now, the synthesis of those OBCPs was obtained through grafting-onto method using Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC). However, complete removal of metal catalyst residues from the resultant copolymer chains is critical and hampers electronic and biomedical applications. In this study, we report an efficient and convenient metal-free click chemistry approach consisting in coupling thiol-containing oligosaccharide blocks to maleimide-terminated polystyrenes. Upon self-assembly in water, spherical micelles of similar size than those obtained by Cu(I)-catalyzed azide-alkyne cycloaddition were formed as evidenced using dynamic light scattering and transmission electron microscopy techniques.

Oxytocin antiserum delays onset of ovarian steroid-induced maternal behavior.

We have previously reported that intracerebroventricular (ICV) administration of oxytocin (OXY) produces a significant increase in maternal behavior in ovariectomized (OVXed) rats given a single priming dose of estrogen. Arginine vasopressin (AVP) has a weaker and more delayed but significant facilitating effect on the onset of maternal behavior. Other investigators have demonstrated that prolonged treatment of OVXed nulliparous rats with estrogen and progesterone followed by withdrawal of progesterone shortens the latency of onset of maternal behavior. We hypothesized that ovarian steroids increase the onset of maternal behavior by a central mechanism involving OXY and possibly AVP. To test this nulliparous Sprague Dawley rats were given SC one Silastic capsule containing 4.4 mg of 17 beta-estradiol eight days after OVX and three capsules each containing 40 mg of progesterone ten days after OVX. Progesterone capsules were removed on the 20th day after OVX, 24 hrs before the introduction of three rat pups (1-5 days old). One hr prior to introduction of pups animals received ICV 10 microliters of anti-oxytocin antiserum (AOA), anti-arginine vasopressin antiserum (AVA), anti-neurotensin antiserum (ANA), normal rabbit serum (NRS), AOA + 250 ng OXY or no ICV infusion. Animals receiving AOA displayed significantly less maternal behavior compared to animals receiving NRS, ANA or no ICV infusion over the first two, six and 25 hrs of pup contact. OXY significantly reversed the inhibitory effect of AOA in the first two hrs of pup contact. AVA significantly decreased the onset of maternal behavior compared to ANA or no ICV infusion over the first six hrs of pup contact.

Erythema elevatum diutinum. Response to dapsone.

A patient with a 20-year history is presented as a classic example of erythema elevatum diutinum, with a dramatic response to dapsone therapy, and with interesting features of improvement during pregnancies and occurrence of giant cell tumors of soft tissue, neither of these latter features having been previously reported.

Ozonolysis of 2'-deoxycytidine: isolation and identification of the main oxidation products.

The ozone-mediated oxidation of 2'-deoxycytidine (dCyd) was investigated on the basis of final product identification. The oxidation reaction gave rise to five major modified nucleosides which were isolated and characterized on the basis of extensive 1H NMR and mass spectrometry measurements. The comparison with the current knowledge of the hydroxyl radical mediated oxidation reactions of 2'-deoxycytidine in aerated aqueous solution, indicates that the formation of ozone oxidation products may be mostly explained by the opening of the pyrimidine C5-C6 double bond. Thus, the formation of the identified products obtained by ozonolysis of 2'-deoxycytidine is accounted for by the initial generation of an ozonide.

Coronary 'Y' stenting: a technique for angioplasty of bifurcation stenoses.

This report describes a case of successful percutaneous coronary transluminal angioplasty (PTCA) of a coronary artery bifurcating lesion using multiple intracoronary stents in an inverted 'Y' configuration. Balloon angioplasty of bifurcation coronary stenoses has a lower procedural success rate, higher restenosis rate and potential for side branch occlusion compared with nonbifurcation lesions. Numerous techniques, including two ('kissing') angioplasty balloons, have been used to overcome these problems. The authors believe that the technique described in this report, inserting stents into both branches simultaneously followed by a third proximal stent in a 'Y' configuration, allows successful dilatation of the bifurcation lesion while preserving flow into both branches and should be considered for similar complex anatomical subsets.

Noncritical disease of the left main coronary artery: limitations of angiography and the role of intravascular ultrasound.

OBJECTIVE: To review the diagnosis and management of noncritical left main stem disease. DATA SOURCES: MEDLINE was searched using the key phrases 'intravascular ultrasound' and 'left main coronary artery'. DATA EXTRACTION: Articles addressing the angiographic or ultrasound assessment of left main stem disease were systematically reviewed. Reference is also made to articles dealing with outcomes for patients with left main stem disease. DATA SYNTHESIS: The assessment of left main stem disease by coronary angiography is often suboptimal. Intravascular ultrasound has demonstrated significant left main stem disease in angiographically normal arteries. It has also demonstrated that mild to moderate left main stem disease frequently accompanies disease in the other coronary arteries. The natural history of mild to moderate left main stem disease has not been well characterized; thus, the optimum management of this condition is not known. CONCLUSIONS: To establish the natural history of noncritical left main stem stenosis, it is proposed that centres performing frequent intravascular ultrasound examinations systematically and quantitatively interrogate the left main stem during all left coronary examinations. Angiographic and ultrasonic measures of left main stem disease can then be correlated with clinical outcome during long term follow-up. This will clearly require the collaboration of several centres to generate a sufficiently large database to address these clinical questions.

Effect of a single bolus of intracoronary basic fibroblast growth factor on perfusion in an ischemic porcine model.

Basic fibroblast growth factor (bFGF) has been shown to induce angiogenesis in various animal models, but the methods of administration used experimentally are not clinically feasible. The objective of this study was to determine whether a single intracoronary bolus injection of bFGF would improve coronary perfusion in a porcine ischemic model that mimics clinical chronic ischemia. A copper coil studded with gold was delivered into the proximal right coronary artery of juvenile Yorkshire pigs and deployed by interventional techniques. After a four-week interval for stenosis maturation, bFGF (100 micrograms) was administered by bolus injection into the left coronary artery in five animals, and vehicle alone was administered in four animals. Angiogenesis and change in right coronary perfusion area were assessed two weeks later by angiography, myocardial contrast echocardiography and immunohistochemistry. The right coronary perfusion area increased significantly after treatment in all but one of the animals that received bFGF but not in any of the controls. Intimal hyperplasia was not induced by bFGF. Capillary density determined histochemically was not different in the two groups. In conclusion, in a porcine ischemic model, bFGF administered by a single bolus intracoronary injection into the contralateral artery improved antegrade perfusion into the ischemic territory although without histological evidence of angiogenesis. This preliminary work merits further investigation.

Induction of a melanoma-specific antibody response by a monovalent, but not a divalent, synthetic GM2 neoglycopeptide.

The GM2 ganglioside represents an important target for specific anticancer immunotherapy. We designed and synthesized a neoglycopeptide immunogen displaying one or two copies of the GM2 tetrasaccharidic moiety. These glycopeptides were prepared using the Huisgen cycloaddition, which enables the efficient ligation of the alkyne-functionalized biosynthesized GM2 with an azido CD4(+) T cell epitope peptide. It is worth noting that the GM2 can be produced on a gram scale in bacteria, which can be advantageous for a scale-up of the process. We show here for the first time that a fully synthetic glycopeptide, which is based on a ganglioside carbohydrate moiety, can induce human tumor cell-specific antibodies after immunization in mice. Interestingly, the monovalent, but not the divalent, form of GM2 peptide construct induced antimelanoma antibodies. Unlike traditional vaccines, this vaccine is a pure chemically-defined entity, a key quality for consistent studies and safe clinical evaluation. Therefore, such carbohydrate-peptide conjugate represents a promising cancer vaccine strategy for active immunotherapy targeting gangliosides.

A high-density, integrated genetic linkage map of lettuce (Lactuca spp.).

An integrated map for lettuce comprising of 2,744 markers was developed from seven intra- and inter-specific mapping populations. A total of 560 markers that segregated in two or more populations were used to align the individual maps. 2,073 AFLP, 152 RFLP, 130 SSR, and 360 RAPD as well as 29 other markers were assigned to nine chromosomal linkage groups that spanned a total of 1,505 cM and ranged from 136 to 238 cM. The maximum interval between markers in the integrated map is 43 cM and the mean interval is 0.7 cM. The majority of markers segregated close to Mendelian expectations in the intra-specific crosses. In the two L. saligna x L. sativa inter-specific crosses, a total of 155 and 116 markers in 13 regions exhibited significant segregation distortion. Data visualization tools were developed to curate, display and query the data. The integrated map provides a framework for mapping ESTs in one core mapping population relative to phenotypes that segregate in other populations. It also provides large numbers of markers for marker assisted selection, candidate gene identification, and studies of genome evolution in the Compositae.

Three-dimensional time-resolved fluorescence imaging by multifocal multiphoton microscopy for a photosensitizer study in living cells.

Two-photon fluorescence microscopy is widely applied to biology and medicine to study both the structure and dynamic processes in living cells. The main issue is the slow acquisition rate due to the point scanning approach limiting the multimodal detection (x, y, z, t). To extend the performances of this powerful technique, we present a time-resolved multifocal multiphoton microscope (MMM) based on laser amplitude splitting. An array of 8 x 8 foci is created on the sample that gives a direct insight of the fluorescence localization. Four-dimensional (4D) imaging is obtained by combining simultaneous foci scanning, time-gated detection, and z displacement. We illustrate time-resolved MMM capabilities for 4D imaging of a photosensitizer inside living colon cancer cells. The aim of this study is to have a better understanding of the photophysical processes implied in the photosensitizer reactivity.

Enhanced fluorescence cell imaging with metal-coated slides.

Fluorescence labeling is the prevailing imaging technique in cell biology research. When they involve statistical investigations on a large number of cells, experimental studies require both low magnification to get a reliable statistical population and high contrast to achieve accurate diagnosis on the nature of the cells' perturbation. Because microscope objectives of low magnification generally yield low collection efficiency, such studies are limited by the fluorescence signal weakness. To overcome this technological bottleneck, we proposed a new method based on metal-coated substrates that enhance the fluorescence process and improve collection efficiency in epifluorescence observation and that can be directly used with a common microscope setup. We developed a model based on the dipole approximation with the aim of simulating the optical behavior of a fluorophore on such a substrate and revealing the different mechanisms responsible for fluorescence enhancement. The presence of a reflective surface modifies both excitation and emission processes and additionally reshapes fluorescence emission lobes. From both theoretical and experimental results, we found the fluorescence signal emitted by a molecular cyanine 3 dye layer to be amplified by a factor approximately 30 when fluorophores are separated by a proper distance from the substrate. We then adapted our model to the case of homogeneously stained micrometer-sized objects and demonstrated mean signal amplification by a factor approximately 4. Finally, we applied our method to fluorescence imaging of dog kidney cells and verified experimentally the simulated results.

Diode-pumped passively mode-locked Nd:YVO4 laser at 914 nm.

We demonstrate, for the first time, to our knowledge, a diode-pumped passively mode-locked Nd:YVO4 laser, operating on the 4F(3/2)-4I(9/2) transition of the neodymium ion at 914 nm. We obtained 8.8 ps pulses at approximately 914 nm at a repetition rate of 94 MHz, and an averaged output power of 87 mW by using a semiconductor saturable absorber mirror.

Fluorescence lifetime imaging with a low-repetition-rate passively mode-locked diode-pumped Nd:YVO4 oscillator.

We report a wide-field fluorescence lifetime imaging microscope based on a low-repetition-rate (3.7-MHz) passively mode-locked diode-pumped laser source. This inexpensive and compact laser source operating in the visible and UV range can excite a wide range of fluorophores of biological interest. We demonstrate that the power of this laser source is highly sufficient for studying biological systems with low quantum yields (autofluorescence of tissues and stained living cells). The maximum measurable lifetime is also strongly increased with this laser source, as fluorescence intensity measurement can occur 250 ns after the excitation pulse.

Differentiation of nearly identical germplasm accessions by a combination of molecular and morphologic analyses.

Polymerase chain reaction (PCR) based random amplified polymorphic DNA (RAPD) markers were used to study the extent of redundancy (duplication of genetic materials) within a genetic resources collection. Nine nearly phenotypically and identical accessions of butterhead lettuce (Lactuca sativa L.) were assayed for their genetic identities. A nonuniform, heterogeneous butterhead line and a crisphead cultivar were added for population comparison. PCR amplification using 13 oligonucleotide primers generated 93 polymorphic bands. The percentage of segregating bands was used to determine within-line variation; values ranged from 0.0 to 12.0%, except for the nonuniform line at 22.6%. Between-line similarity was measured using similarity coefficients and ranged from 0.919 to 0.985. The relationship between the crisphead accession and a composite of all butterhead accessions was 0.84. Selfed progeny of each line were measured for morphological uniformity. The variation obtained from these biological data was compared with variation detected at the DNA level and each was positively correlated. Results demonstrate that RAPD analyses may serve as a major source of information for separation of closely related accessions, especially when integrated with phenotypic measures.

A factor released from coronary vascular endothelium inhibits myocardial contractile performance.

The effect of continuous intracoronary infusion of bradykinin (BK; 10(-6) M) on the myocardial contractile performance of isolated Langendorff-perfused ferret hearts was investigated. BK produced a transient but marked inhibition of contractile performance, the changes being fully resolved by 2-3 min. This effect of BK was blocked after disruption of the coronary vascular endothelium with Triton X-100 (0.5%). Prior infusion of the endothelium-derived relaxing factor (EDRF) synthesis inhibitor N omega-nitro-L-arginine (10(-4) M) failed to alter the mechanical effects induced by BK. These changes in contractile performance were reduced, however, by the K+ channel-blocking agent glibenclamide (10(-6) M) but were unaffected by tetrapentylammonium bromide (5 x 10(-5) M) or 4-aminopyridine (10(-3) M). These findings therefore indicate that BK inhibits myocardial contractile performance by opening myocardial ATP-sensitive K+ channels, probably via the release of an endothelium-derived factor unrelated to EDRF.

Three-dimensional acousto-optic imaging in biological tissues with parallel signal processing.

An original acousto-optic method is described that allows one to reveal optical contrasts through biological tissues that are several centimeters thick with a millimeter-sized resolution. This technique is based on the interaction of scattered laser light with a focused ultrasonic field. The modulation depth of the optical speckle is related to local optical properties of the sample. Our parallel-processing approach to the demodulation of the speckle improves the observed degree of modulation by 2 orders of magnitude and quickly yields a good statistical value. Optically absorbing objects were imaged inside 35-mm-thick biological tissues.

Prolonged pharyngeal impaction of dentures following stroke.

Dysphagia is a well-recognized complication of stroke. We report two cases of dysphagia in stroke patients caused by the pharyngeal impaction of dental prostheses. Radiologic identification of such impaction is unreliable due to the increasing use of radiolucent material in dental prostheses. We recommend direct or indirect laryngoscopy to exclude foreign body impaction in all patients complaining of dysphagia.

Synthesis and evaluation as glycosidase inhibitors of 2,5-imino-D-glucitol and 1,5-imino-D-mannitol related derivatives.

Selectively functionalized 2,5-imino-D-glucitol and 1,5-imino-D-mannitol derivatives were synthesized and tested as precursors of hydrolytically resistant pseudo-disaccharides. Among them N-acetyl-6-amino-6-deoxy-2,5-imino-D-glucitol (11) and N-acetyl-6-amino-6-deoxy-1,5-imino-D-mannitol (12) were found potent and specific inhibitors against beta-D-glucosidase and alpha-L-fucosidase, respectively.