Kinetics of DSB rejoining and formation of simple chromosome exchange aberrations

PURPOSE: To investigate the role of kinetics in the processing of DNA double strand breaks (DSB), and the formation of simple chromosome exchange aberrations following X-ray exposures to mammalian cells based on an enzymatic approach. METHODS: Using computer simulations based on a biochemical approach, rate-equations that describe the processing of DSB through the formation of a DNA-enzyme complex were formulated. A second model that allows for competition between two processing pathways was also formulated. The formation of simple exchange aberrations was modelled as misrepair during the recombination of single DSB with undamaged DNA. Non-linear coupled differential equations corresponding to biochemical pathways were solved numerically by fitting to experimental data. RESULTS: When mediated by a DSB repair enzyme complex, the processing of single DSB showed a complex behaviour that gives the appearance of fast and slow components of rejoining. This is due to the time-delay caused by the action time of enzymes in biomolecular reactions. It is shown that the kinetic- and dose-responses of simple chromosome exchange aberrations are well described by a recombination model of DSB interacting with undamaged DNA when aberration formation increases with linear dose-dependence. Competition between two or more recombination processes is shown to lead to the formation of simple exchange aberrations with a dose-dependence similar to that of a linear quadratic model. CONCLUSIONS: Using a minimal number of assumptions, the kinetics and dose response observed experimentally for DSB rejoining and the formation of simple chromosome exchange aberrations are shown to be consistent with kinetic models based on enzymatic reaction approaches. A non-linear dose response for simple exchange aberrations is possible in a model of recombination of DNA containing a DSB with undamaged DNA when two or more pathways compete for DSB repair.     

Simulation of 125I-based radioprobing experiments to study DNA quadruplex structure and topology

Purpose: Certain guanine-rich DNA sequences have the capacity to fold into four-stranded structures stabilized by the stacking of square planar arrangements of four hydrogen-bonded guanine bases. However both the overall topology of folding and the more detailed three dimensional structure of these quadruplexes is difficult to determine or predict, and they can be polymorphic, altering radically depending on environmental conditions. Radioprobing experiments, in which Auger electrons emitted during the decay of a ¹²⁵I-containing base induce strand cleavage in a distance- and structure-dependent manner, have provided possible means of determining these details. Here we have used a combination of computer simulation methods to study the information obtained by one such experiment, reported in 2004.

Method: Models were constructed of three quadruplex topologies considered in the experiment, and one other topology proposed more recently. Molecular Dynamics simulations were used to equilibrate these structures and monitor how they evolved over several nanoseconds in solution. Snapshots from the trajectories were then subjected to Monte Carlo track structure prediction, from which theoretical cleavage patterns have been extracted.

Results: The four topologies were found to yield quite different cleavage patterns, which allow the presence of particular conformations in an experiment to be predicted.

Conclusion: Radioprobing, which is usable in biologically relevant environments, is sensitive enough to distinguish with some confidence between alternative folding topologies in a DNA structure. Monte Carlo track structure simulation can reinforce or question conclusions drawn from experiment, and Molecular Dynamics used with various restraints provides a practical means of guiding a model towards one that yields cleavage patterns closer to those found experimentally.     

Esophageal Aperistalsis and Gastroesophageal Reflux Disorder: Return of Peristalsis after H2-Blocker Therapy

Objectives: Distal esophageal aperistalsis has rarely been reported among patients with gastroesophageal reflux disorder. The purpose of this study, therefore, was to address the frequency with which disorders of peristalsis in general-and distal esophageal aperistalsis in particular-occur in adults with gastroesophageal reflux disorder. Patients and Methods: We studied 314 patients who were referred to our gastrointestinal motility laboratory. On the basis of the endoscopic data, they were divided into three groups: group I, symptomatic patients without endoscopic esophagitis; group II, patients with mild endoscopic esophagitis; and group III. patients with erosive esophagitis. An age-matched group of patients with chest pain unrelated to reflux served as the control. Results: Some form of peristaltic dysfunction was recorded in 56% of the patients with gastroesophageal reflux disorder, significantly more than in the control group ( p < 0.01). A significant correlation existed between the esophageal motor dysfunction scores and the severity of reflux disease. Distal esophageal aperistalsis was present in 3.1% of the reflux groups. There was a correlation between severity of reflux disease and the prevalence of aperistalsis. Aperistalsis occurred in none of the patients in group I, in 3.6% of group II, and in 12.5% of group III ( p < 0.0001). Seven of the patients with aperistalsis who has been treated with H2-blockers were reexamined 4 months later. Return of peristalsis was seen in three of them. Conclusion: Esophageal aperistalsis can be seen in a minority of patients with severe gastroesophageal reflux disorder and is probably a reversible condition.     

Kinetics of DSB rejoining and formation of simple chromosome exchange aberrations

Purpose : To investigate the role of kinetics in the processing of DNA double strand breaks (DSB), and the formation of simple chromosome exchange aberrations following X-ray exposures to mammalian cells based on an enzymatic approach. Methods : Using computer simulations based on a biochemical approach, rate-equations that describe the processing of DSB through the formation of a DNA-enzyme complex were formulated. A second model that allows for competition between two processing pathways was also formulated. The formation of simple exchange aberrations was modelled as misrepair during the recombination of single DSB with undamaged DNA. Non-linear coupled differential equations corresponding to biochemical pathways were solved numerically by fitting to experimental data. Results : When mediated by a DSB-repair enzyme complex, the processing of single DSB showed a complex behaviour that gives the appearance of fast and slow components of rejoining. This is due to the time-delay caused by the action time of enzymes in biomolecular reactions. It is shown that the kinetic- and dose- responses of simple chromosome exchange aberrations are well described by a recombination model of DSB interacting with undamaged DNA when aberration formation increases with linear dose-dependence. Competition between two or more recombination processes is shown to lead to the formation of simple exchange aberrations with a dose-dependence similar to that of a linear-quadratic model. Conclusions : Using a minimal number of assumptions, the kinetics and dose-response observed experimentally for DSB rejoining and the formation of simple chromosome exchange aberrations are shown to be consistent with kinetic models based on enzymatic reaction approaches. A non-linear dose-response for simple exchange aberrations is possible in a model of recombination of DNA containing a DSB with undamaged DNA when two or more pathways compete for DSB repair.     

Future directions of laser phototherapy for diagnosis and treatment of malignancies: Fantasy,fallacy, or reality?

A new and highly promising adjunctive modality for the diagnosis and therapy of malignancies is under development using lasers and tumor targeting dyes. To reach the eventual goal of clinical treatment, several current “fantasies and fallacies” regarding laser applications in medicine must be identified and their problems clearly outlined. A multidisciplinary scientific approach is also required to enable the clinical practicality of this laser targeting approach. Many new dyes and laser wavelengths are being tested to improve specific tumor uptake and/or retention, lower systemic toxicity, increase tissue penetration, and identify fluorochromes with synergistic properties to further enhance laser tumoricidal effects. Rapid technological advancements in magnetic resonance imaging may now provide an extremely sensitive way to detect and monitor laser-tissue effects, and allow efficient interstitial laser phototherapy of deep and sometimes inaccessible tumors. The current and future prospectives of the emerging field of laser phototherapy are described.     

Off-pump coronary artery grafting in patients with left main coronary artery disease

BACKGROUND: Left main coronary artery disease (LMCD) is considered a relative contraindication to off-pump coronary artery bypass (OPCAB) grafting. This study evaluates the safety and feasibility of OPCAB in these patients compared to an on-pump group (cardiopulmonary bypass, CPB) with LMCD. METHODS: Between January 2000 and July 2002, 95 patients with left main coronary artery stenosis >50% underwent coronary revascularization. Seventy-three unselected patients underwent OPCAB and 22 underwent revascularization using CPB. The techniques used for OPCAB included the use of deep traction sutures in the posterior pericardium and stabilizers to expose the distal coronary targets. Intraluminal coronary shunts were routinely used during construction of the anastomoses. Variables were analyzed using a Student's paired t-test with statistical significance defined as p < 0.05. RESULTS: The mean age in the OPCAB group was 59.9 years and the CPB group 61.8 years (p = 0.54). There were 56 males (77%) in the OPCAB and 18 (82%) in the CPB groups. Mean preoperative left ventricular ejection fraction (LVEF) was 40.3% in OPCAB and 47.3% in CPB (p = 0.015). Average number of grafts was 3.1 in OPCAB and 4.1 in CPB (p = 0.0038). There were no conversions to CPB in those patients initially chosen to undergo OPCAB. There were no early deaths in OPCAB. There was one death in CPB. Mean hospital length of stay was 6.9 days for OPCAB and 9.1 for CPB (p = 0.0159). CONCLUSIONS: Patients with LMCD can undergo OPCAB grafting safely and effectively despite reduced LVEF. LMCD should no longer be seen as a contraindication to perform OPCAB grafting.     

Calculated DNA damage from gadolinium Auger electrons and relation to dose distributions in a head phantom

PURPOSE: To calculate the number of 157Gadolinium (157Gd) neutron capture induced DNA double strand breaks (DSB) in tumor cells resulting from epithermal neutron irradiation of a human head when the peak tissue dose is 10 Gy. To assess the lethality of these Gd induced DSB. MATRIALS AND METHODS: DNA single and double strand breaks from Auger electrons emitted during 157Gd(n,gamma) events were calculated using an atomistic model of B-DNA with higher-order structure. When combined with gadolinium neutron capture reaction rates and neutron and photon physical dose rates calculated from the radiation transport through a model of the human head with explicit tumors, peak tissue dose can be related to the number of Auger electron induced DSB in tumor cell DNA. The lethality of these DNA DSB were assessed through a comparison with incorporated 125I decay cell survival curves and second comparison with the number of DSB resulting from neutron and photon interactions. RESULTS: These calculations on a molecular scale (microscopic calculations) indicate that for incorporated 157Gd, each neutron capture reaction results in an average of 1.56 +/- 0.16 DNA single strand breaks (SSB) and 0.21 +/- 0.04 DBS in the immediate vicinity (approximately 40 nm) of the neutron capture. In an example case of Gd Neutron Capture Therapy (GdNCT), a 1 cm radius midline tumor, peak normal tissue dose of 10 Gy, and a tumor concentration of 1000 ppm Gd, result in a maximum of 140 +/- 27 DSBs per tumor cell. CONCLUSIONS: The number of DSB from the background radiation components is one order of magnitude lower than the Gd Auger electron induced DSB. The cell survival of mammalian cell lines with a similar amount of complex DSB induced from incorporated 125I decay yield one to two magnitudes of cell killing. These two points indicate that gadolinium auger electrons could significantly contribute to cell killing in GdNCT.     

Correlation of cephalic anthropometric parameters and microsurgical anatomy of the optic nerves: a cadaveric morphometric study

BACKGROUND: This study was designed to elucidate the possible correlation of cranial anthropometric measurements with the chiasm to limbus sphenoidale distance to facilitate preoperative estimation of this distance and to choose a better surgical approach. METHODS: Thirty-three fresh adult cadaver heads (22 males and 11 females) were evaluated for cranial anthropometric measurements. The precraniotomy anthropometric measurements included (A) inion to nasion distance and (B) the longest intermeatal meridian. Subsequently, with a standard craniotomy, the following intervals were measured: (C) optic chiasm to falciform ligament, (D) anterior aspect of optic chiasm to limbus sphenoidale, and (E) limbus sphenoidale to inner nasion. A combined ratio parameter, labeled as (F), was calculated from the following equation: F = B/E x 10. RESULTS: The mean values and standard errors of the mean of parameters A to F were 195.8 +/- 14.53 mm, 374.7 +/- 25.29 mm, 10.47 +/- 1.89 mm, 9.93 +/- 2.01 mm, 38.46 +/- 3.17 mm, and 9.81 +/- 1.11, respectively. The parameter D had significant correlation to the parameters B, C, E, and F. The most significant correlation was seen between parameters D and F (p < 0.001). According to linear regression assessment between parameters D and F, the following regression equation was obtained: D = 4.24 + 0.58F. CONCLUSIONS: Optic nerve topography and dimensions show inter-personal variations that may be anticipated to some extent with cranial anthropometric data. Calculating of F ratio gives us an acceptable estimation of the actual distance of chiasm to limbus sphenoidale, which in turn can help the surgeon to select the approach to tumors of intrasellar region. However, the role of meticulous imaging studies cannot be overemphasized to confirm the anticipated estimations.