Discriminating Intercalative Effects of Threading Intercalator Nogalamycin, from Classical Intercalator Daunomycin, Using Single Molecule Atomic Force Spectroscopy.

DNA threading intercalators are a unique class of intercalating agents, albeit little biophysical information is available on their intercalative actions. Herein, the intercalative effects of nogalamycin, which is a naturally-occurring DNA threading intercalator, have been investigated by high-resolution atomic force microscopy (AFM) and spectroscopy (AFS). The results have been compared with those of the well-known chemotherapeutic drug daunomycin, which is a non-threading classical intercalator bearing structural similarity to nogalamycin. A comparative AFM assessment revealed a greater increase in DNA contour length over the entire incubation period of 48 h for nogalamycin treatment, whereas the contour length increase manifested faster in case of daunomycin. The elastic response of single DNA molecules to an externally applied force was investigated by the single molecule AFS approach. Characteristic mechanical fingerprints in the overstretching behaviour clearly distinguished the nogalamycin/daunomycin-treated dsDNA from untreated dsDNA-the former appearing less elastic than the latter, and the nogalamycin-treated DNA distinguished from the daunomycin-treated DNA-the classically intercalated dsDNA appearing the least elastic. A single molecule AFS-based discrimination of threading intercalation from the classical type is being reported for the first time.

Interactions of Histone Acetyltransferase p300 with the Nuclear Proteins Histone and HMGB1, As Revealed by Single Molecule Atomic Force Spectroscopy.

One of the important properties of the transcriptional coactivator p300 is histone acetyltransferase (HAT) activity that enables p300 to influence chromatin action via histone modulation. p300 can exert its HAT action upon the other nuclear proteins too--one notable example being the transcription-factor-like protein HMGB1, which functions also as a cytokine, and whose accumulation in the cytoplasm, as a response to tissue damage, is triggered by its acetylation. Hitherto, no information on the structure and stability of the complexes between full-length p300 (p300FL) (300 kDa) and the histone/HMGB1 proteins are available, probably due to the presence of unstructured regions within p300FL that makes it difficult to be crystallized. Herein, we have adopted the high-resolution atomic force microscopy (AFM) approach, which allows molecularly resolved three-dimensional contour mapping of a protein molecule of any size and structure. From the off-rate and activation barrier values, obtained using single molecule dynamic force spectroscopy, the biochemical proposition of preferential binding of p300FL to histone H3, compared to the octameric histone, can be validated. Importantly, from the energy landscape of the dissociation events, a model for the p300-histone and the p300-HMGB1 dynamic complexes that HAT forms, can be proposed. The lower unbinding forces of the complexes observed in acetylating conditions, compared to those observed in non-acetylating conditions, indicate that upon acetylation, p300 tends to weakly associate, probably as an outcome of charge alterations on the histone/HMGB1 surface and/or acetylation-induced conformational changes. To our knowledge, for the first time, a single molecule level treatment of the interactions of HAT, where the full-length protein is considered, is being reported.

Biodegradable and biocompatible soy protein/polymer/adhesive sticky nano-textured interfacial membranes for prevention of esca fungi invasion into pruning cuts and wounds of vines.

Adhesive biodegradable membranes (patches) for the protection of pruning locations of plants from esca fungi attacks were developed using electrospun soy protein/polyvinyl alcohol and soy protein/polycaprolactone nanofibers. Several different water-soluble adhesives were either added directly to the electrospinning solutions or electrosprayed onto the as-spun nanofiber mats. The nanofibers were deposited onto a biodegradable rayon membrane, and are to be pressed onto the pruned location on a plant. The pore size in the nanofiber mats is sufficient for physically blocking fungi penetration, while the outside rayon membrane provides sufficient mechanical support in handling prior to deposition on a plant. Diseases like Vine Decline are one of the most important cases where such a remedy would be needed. It should be emphasized that these novel biodegradable and sticky patches are radically different from the ordinary electrospun ultra-filtration membranes. The normal and shear specific adhesive energy of the patches were measured, and the results show that they can withstand strong wind without being blown off. On the other hand, the patches possess sufficient porosity for plant breathing.

Enhanced foamability of sodium dodecyl sulfate surfactant mixed with superspreader trisiloxane-(poly)ethoxylate.

Gravitational drainage from thin vertical surfactant solution films and gravitational drainage in a settler column are used to study the behavior of foams based on two-surfactant mixtures. Namely, solutions of the anionic sodium dodecyl sulfate (SDS) and nonionic superspreader SILWET L-77, and their mixtures at different mixing ratios, are studied. It is shown, for the first time, that solutions having a longer lifetime in the vertical film drainage process also possess a higher foamability. An additional and unexpected unique result is that when using a mixed surfactant system, the foamability can be much greater than the foamabilities of the individual components.

Superspreaders versus "cousin" non-superspreaders: disjoining pressure in gravitational film drainage.

Gravitational drainage of vertical films supported on a wire frame of two superspreaders SILWET L-77 and BREAK-THRU S 278 and their respective "cousin" non-superspreaders SILWET L-7607 and BREAK-THRU S 233 revealed drastic differences. The superspreader films showed complicated dynamic "turbulent"-like interferometric patterns in distinction from the ordered color bands of the "cousin" non-superspreaders, which resembled those of the ordinary surfactants. Nevertheless, the superspreader films stabilized themselves at the thickness about 35 nm and revealed an order of magnitude longer lifetime before bursting compared to that of the "cousin" non-superspreaders. Notably, the superspreaders revealed drastic differences from the non-superspreaders in aqueous solutions with no contact with any solid hydrophobic surface. The self-stabilization of the superspreader films is attributed to significant disjoining pressure probably related to long superspreader bilayers hanging from the free surfaces. The scaling law for the disjoining pressure was found as p(disj)(h) ~ h(-m) (with m ≈ 9-11) for the sufficiently concentrated superspreader solutions, and as p(disj)(h) ~ h(-s) (with s ≈ 6) for more dilute solutions (in both cases, concentrations were above the critical micelle concentration). The non-superspreaders do not possess any significant disjoining pressure even in the films with thicknesses in the 35-100 nm range. The results show that gravitational drainage of vertical films is a useful simple tool for measuring disjoining pressure.

Gravitational drainage of foam films.

Gravitational drainage from thick plane vertical soap films and hemispherical bubbles is studied experimentally and theoretically. The experiments involve microinterferometry kindred to the one used in the experiments in the Scheludko cell. The following surfactants were used in the experiments: cationic dodecyltrimethylammonium bromide (DTAB), anionic sodium dodecyl sulfate (SDS), anionic Pantene shampoo which primarily contains sodium lauryl sulfate, nonionic tetraethylene glycol monooctyl ether (C8E4), and nonionic Pluronic (P-123) surfactants at different concentrations. The theoretical results explain the drainage mechanism and are used to develop a new method of measurement of the surface elasticity and to test it on the above-mentioned surfactants.

The role of laser radiation therapy in maxillary sinusitis.

Efficacy of prescribed noninvasive & invasive types of treatment of maxillary sinusitis has been compared with low-dose LASER therapy (LLT). After going through the observations of different authors on the therapeutic role of LLT (GaA1AS-LASER) in non-ENT infective diseases, its use in 'sinusitis' has been adjudged. Such type of study-report has not been found by us in the literatures, available to us.It was observed here that the LLT has produced better result with better acceptation by patients in the management of 'sinusitis' A LLT-treatment virtually produced no side-effect/complication as compared with the invasive methods.In this study. 25 cases, treated by LLT, out of 200 cases, treated by conventional methods, yielded good result in 22 & satisfactory result in 3 cases.

Interrupted right aortic arch and origin of the left pulmonary artery from the aorta in DiGeorge syndrome.

We describe a neonate with DiGeorge syndrome undergoing diagnosis and successful repair of interrupted right aortic arch and origin of the left pulmonary artery from the aorta. We discuss a link between this lesion and persistence of a left fifth arch.

Effect of purified glutaminase from human ascites fluid on experimental tumor bearing mice.

Glutamine is the major respiratory fuel and energy source of the rapidly proliferating tumor cells and that is why glutamine clearance by glutaminase therapy provides an opportunity to fight against the neoplasm. Glutaminase from bacterial source was tried on experimental models but had to be excluded because of its limited efficacy. Search for a better glutaminase continued exploiting the mammalian sources. In the present study, glutaminase purified from human ovarian cancer ascites fluid was used in experimental solid and ascites mice model alone and in combination with Cu-Sulphate and heparin. Cumulative findings indicate that the enzyme alone is quite effective in lowering tumor burden and reducing not only the tumor induced angiogenesis, but also an angiogenic inducer, heparin mediated angiogenesis. However, the presence of Cu with the enzyme, amplified the antineoplastic response by improving anti-angiogenic potential and hematological status of the tumor bearing host. Therefore, Cu-glutaminase combination strengthened the hypothesis that together they may provide a better therapeutic regimen in experimental mice tumor model.

Avirulent mutants of Macrophomina phaseolina and Aspergillus fumigatus initiate infection in Phaseolus mungo in the presence of phaseolinone; levamisole gives protection.

To evaluate the role of phaseolinone, a phytotoxin produced by Macrophomina phaseolina, in disease initiation, three nontoxigenic avirulent mutants of the fungus were generated by UV-mutagenesis. Two of them were able to initiate infection in germinating Phaseolus mungo seeds only in the presence of phaseolinone. The minimum dose of phaseoli-none required for infection in 30% seedlings was 2 5 mg/ml. A human pathogen, Aspergillus fumigatus was also able to infect germinating seeds of P. mungo in the presence of 5 mg/ml concentration of phaseolinone. Phaseolinone seemed to facilitate infection by A. fumigatus, which is not normally phytopathogenic, by reducing the immunity of germinating seedlings in a nonspecific way. Levamisole, a non-specific immunopotentiator gave protection against infection induced by A. fumigatus at an optimum dose of 50 mg/ml. Sodium malonate prevented the effects of levamisole.

The evolution of ductus arteriosus treatment.

The treatment of patent ductus arteriosus (PDA) has evolved over the years. We reviewed 231 non-premature children (group 1) undergoing surgical closure of a PDA between January 1985 and December 1997, and 30 children (group 2) undergoing transcatheter closure from May 1995 to December 1998. The median age and weight at operation in group 1 were 13 months (range, 0.5-174 months) and 9.5 kg (range, 1.9-49.7 kg), respectively. There was one intra-operative death (0.4%) secondary to hemorrhage. Immediate extubation was performed in 208 patients (90%). Intra-operative chest tube use decreased from 73.3% to 10% between the 1985-88 and 1996-97 periods (P < 0.001). Postoperative pneumothoraces occurred in 33/131 (25%) patients with only one patient (0.7%) requiring drainage. Eleven patients had complications including wound infection in four, vocal cord paralysis in three, and left pulmonary artery stenosis in one. The median length of stay (LOS) was 5 days (range, 2-43 days). Follow-up echocardiogram was performed in 146/230 patients (63%) and revealed a residual PDA in six (4%); two being re-ligated, two remaining clinically insignificant, and two spontaneously resolved at 7 and 28 months follow-up. The remaining 84 patients had no clinical signs of a residual PDA. In group 2, where a transcatheter coil occlusion technique was used, the median age and weight at procedure were 31 months (range, 9-320 months) and 14.9 kg (range, 9-69.7 kg), respectively. Vascular complications occurred in four patients (13.3%). One patient developed hemolysis and hemoglobinuria requiring hospital admission. Four patients required a second intervention. At the most recent echocardiographic assessment, four patients (13.3%) had a residual PDA.

Hemostatic parameters and platelet activation marker expression in cyanotic and acyanotic pediatric patients undergoing cardiac surgery in the presence of tranexamic acid.

We have investigated hemostatic parameters including platelet activation in 56 pediatric patients with or without cyanosis undergoing cardiopulmonary bypass (CPB) and cardiac surgery to repair congenital defects. Patients were participants in a study assessing the effects of tranexamic acid on surgery-related blood loss. Parameters monitored included blood loss, prothrombin F1.2, thrombin-antithrombin complexes, t-PA, PAI-1, plasminogen, fibrin D-dimer, and plasma factor XIII. Additionally, flow cytometry monitored platelet degranulation (P-selectin or CD63), as well as surface-bound fibrinogen, von Willebrand factor and factor XIIIa. Cyanotic patients had evidence of supranormal coagulation activation as both fibrin D-dimer and PAI-1 levels were elevated prior to surgery. While the extent of expression of P-selectin or CD63 was not informative, platelet-associated factor XIIIa was elevated in cyanotic patients at baseline. In both patient groups, CPB altered platelet activation state and coagulation status irrespective of the use of tranexamic acid.

Laryngotracheal stenosis & pharyngocutaneous fistula in cut-throat injuries - how we managed them.

Two successfully treated cases, one of laryngotracheal stenosis & another of pharyngocutaneous fistula have been presented. These complications developed due to bad initial management & poor physical health following cut-throat Injury, It was observed that cheaper stents like polythelene endotracheal tube gave good remits like costlier stents in the treatment of stenosis. Mild degree of subglottic stenosis could he dilated to yield good airway thus avoiding major surgery.It should be remembered that proper initial management & early repair of the 'cut-throat injury' would avert these complications.Post-operative follow-up for at least a year & more is mandatory to assess the final outcome.

Extracardiac Fontan operation with tube fenestration allowing transcatheter coil occlusion.

A fenestration may improve the immediate postoperative course after a Fontan procedure by preserving the cardiac output. We describe a simple and safe technique of fenestration amenable to coil occlusion, which can be carried out in most cardiac catheterization laboratories.

Intermediate follow-up of right ventricular outflow tract reconstruction with allograft conduits.

BACKGROUND: Allograft conduits are among many varieties of material used for right ventricular outflow tract reconstruction. They invariably need to be replaced due to growth of the patient or conduit failure. METHODS: From June 1984 to June 1996, a total of 76 patients underwent reconstruction of the right ventricular outflow tract with an allograft conduit: 51 aortic and 25 pulmonary. The median age, weight and conduit size at surgery were 37 months (range, 0.2 to 228 months), 12.4 kg (range, 2.9 to 61.4 kg), and 17 mm (range, 8 to 26 mm), respectively. RESULTS: The hospital mortality was 5.3% (4 of 76 patients) and 2 patients died at 9 and 78 months follow-up. The median follow-up was 61 months (range, 2 to 132 months). Reoperation was necessary in 22 patients (28.9%) at a median interval of 50.5 months (range, 3 to 109 months) and the median conduit size was 21 mm (range, 12 to 23 months). There was no mortality. Freedom from reoperation at 64 months was 49.5% for conduits 15 mm and smaller, and 73.3% for conduits 16 mm and larger. Analysis by age shows freedom from reoperation at 64 months of 49.4% and 74.5% for patients younger than and older than 2 years, respectively. At 54 months there was no statistical difference in freedom from reoperation between pulmonary and aortic allografts. CONCLUSION: Right ventricular outflow tract reconstruction with allograft conduits results in a high reoperation rate at 4 years but provides significantly longer freedom from reoperation with conduits larger than 15 mm or in patients over 24 months of age.

Risks of repeat sternotomy in pediatric cardiac operations.

BACKGROUND: Repeat sternotomy has been associated with increased perioperative risks. METHODS: We reviewed 165 patients undergoing 192 repeat sternotomies between January 1985 and January 1997 (group 1) and a control group of 184 patients (group 2). The operations in group 1 were valve procedures in 94 patients, Fontan procedure in 46, ventricular septal defect closure in 10, pulmonary arterioplasty in 17, and others in 25; in group 2 ventricular or atrial septal defect closure in 120 patients, tetralogy of Fallot repair in 26, valve procedures in 16, bidirectional Glenn anastomosis in 7, repair of transposition of the great arteries in 7, pulmonary arterioplasty in 4, and others in 4. RESULTS: The hospital mortality was 2.6% in group 1 and 3.8% in group 2. Cardiac laceration occurred in 10 of 192 patients (5.2%), requiring emergent femorofemoral bypass in 6 patients. Two patients sustained an air embolism that was successfully treated with a hyperbaric chamber. Median total blood loss and requirements were not significantly different between the two groups. The length of stay in the intensive care unit and in the hospital were 4 days (range, 1 to 80 days) and 11 days (range, 1 to 135 days) in group 1, and 2 days (range, 1 to 87 days) and 7 days (range, 1 to 131 days) in group 2 (p < 0.02 and p < 0.002, respectively). The rate of complications was not significantly different in group 1 versus group 2. Overall survival was 97% (group 1) and 95% (group 2) at 120 months' follow-up (not significant). CONCLUSIONS: With careful surgical technique and judicious use of femorofemoral bypass, the risk of repeat sternotomy is minimized.

Is surgical closure of patent ductus arteriosus a safe procedure in premature infants?

Despite indomethacin therapy, many premature infants require surgical closure of their patent ductus arteriosus (PDA). Between January 1985 and December 1997, 176 premature infants underwent surgical closure of PDA by vascular clip after failure of medical treatment. The median gestational age and birth weight were 26 weeks (range 23-36 weeks) and 847.5 g (range 400-2300 g), respectively. The median age at diagnosis and at surgery was 4 days (range, 1-37) and 21 days (range, 4-60) respectively. The median weight at surgery was 982.5 g (range 475-2740 g). Of these infants, 168 (95%) were intubated prior to surgery and the median time to extubation was 21 days (range 1-273 days). There were no operative deaths but 11 infants (6.4%) died from complications of prematurity (sepsis, bronchopulmonary dysplasia and pulmonary hemorrhage). The frequency of chest tube insertion at surgery decreased from 41.7% to 10% between the 1985-88 and 1996-97 periods (P<0.01). Three infants (1.7%) developed vocal cord paralysis directly related to the position of the vascular clip. Echocardiography confirmed PDA closure in 43 infants (24.4%) while the remaining 133 had no clinical signs of PDA. Surgical closure of PDA by vascular clip carries a very low morbidity in premature infants.

Effect of glutamate-aspartate reperfusion on postischemic neonatal myocardium.

OBJECTIVE: We postulated that L-glutamate- and L-aspartate-enriched perfusate would improve functional recovery of postischemic neonatal rabbit hearts. METHODS: Isolated working neonatal rabbit hearts were perfused with Krebs-Henseleit buffer and then subjected to 1 hour of hypothermic cardioplegic arrest with St. Thomas' Hospital solution. Hearts were then reperfused with L-glutamate- and L-aspartate-enriched (20 mmol/L) Krebs-Henseleit buffer (AA-enriched Krebs-Henseleit buffer). Hearts reperfused with Krebs-Henseleit buffer alone acted as controls (experiment A). Another group of hearts underwent a similar protocol but were reperfused with the AA-enriched Krebs-Henseleit buffer with correction of the sodium content (experiment B). RESULTS: Hearts reperfused with AA-enriched Krebs-Henseleit buffer showed a significant decrease in aortic flow at both 15 (p = 0.04) and 30 (p = 0.025) minutes compared with controls. Arrhythmias were frequent. Sodium content of the AA-enriched Krebs-Henseleit buffer was 174 +/- 0.5 mmol/L. In experiment B, hearts reperfused with the AA-enriched Krebs-Henseleit buffer with correction of the sodium content exhibited no difference in aortic flow and cardiac output at either 15 or 30 minutes (p = 0.95 and 0.5 and 0.48 and 0.78, respectively) compared with controls. No arrhythmias were observed. The sodium content of the AA-enriched Krebs-Henseleit buffer was 146 +/- 0.7 mmol/L. CONCLUSIONS: A beneficial effect on functional recovery of neonatal hearts reperfused with AA-enriched Krebs-Henseleit buffer was not demonstrated.

The effect of preoperative tranexamic acid on blood loss after cardiac operations in children.

Children undergoing cardiac operations in which cardiopulmonary bypass is used are at risk of significant postoperative blood loss. The acquired coagulopathy is complex but is thought to be due, in part, to excessive fibrinolysis. We examined the possibility of reducing postoperative blood loss in children by using the antifibrinolytic drug tranexamic acid. Using a prospective, randomized, double-blind study design, we administered a single dose of tranexamic acid (50 mg/kg intravenously) or saline placebo, before skin incision, in 88 children undergoing cardiac operations. Post-operative blood loss and fluid replacement were recorded for the next 24 hours. In addition, hemoglobin, platelet counts, and coagulation measures were recorded every 6 hours. When all patients were examined, there was no significant difference in postoperative blood loss between the treated and placebo groups (21.2 +/- 12 ml/kg per 24 hours, tranexamic acid, vs 27.2 +/- 20.3 mls/kg per 24 hours, placebo). However, when the children with cyanosis were analyzed separately, there was a highly significant difference in blood loss between the groups during the first 6 hours (11.2 +/- 3.7 ml/kg per 6 hours, tranexamic acid, vs 27.2 +/- 11.4 mls/kg per 6 hours, placebo; p < 0.002), as well as the overall 24 hour study period (23.7 +/- 7.5 mls/kg per 24 hours, tranexamic acid, vs 48.9 +/- 27.6 mls/kg per 24 hours, placebo; p < 0.02). Also significantly less blood and blood products were administered to the treated cyanosed group. Tranexamic acid produced a significant reduction in postoperative blood loss and blood product requirements in children with cyanosis undergoing heart operations. The drug had no effect in children without cyanosis or those requiring a second thoracotomy.

ICAM-1 and VCAM-1 expression in accelerated cardiac allograft arteriopathy and myocardial rejection are influenced differently by cyclosporine A and tumour necrosis factor-alpha blockade.

Expression of the vascular cell adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) occurs in allograft myocardium and in coronary arteries, promoting adhesion and transendothelial migration of inflammatory cells. We therefore investigated, in cholesterol-fed rabbits 9-10 days following heterotopic cardiac transplantation, whether the reduction of both myocardial rejection and graft arteriopathy with cyclosporine A (CsA) or graft arteriopathy alone with tumour necrosis factor-alpha soluble receptor (TNFsr) was associated with suppression of ICAM-1 and VCAM-1 expression. Host hearts showed negative immunostaining for these adhesion molecules, whereas donor specimens from untreated (control) rabbits showed moderate immunostaining for ICAM-1 and weaker immunostaining for VCAM-1 in the coronary arteries, myocardium (cardiac myocytes), and perivenular regions. The selective reduction of the coronary arteriopathy with TNFsr was associated with somewhat reduced expression of these adhesion molecules in the arteries, whereas CsA also suppressed myocardial rejection and markedly decreased both vascular and myocyte expression of ICAM-1 and VCAM-1.