Biomedical subjects

Eugene Demidenko

Publications and source records attributed to Eugene Demidenko.

At least 19 recordsLinked to original sources

PubMed2007

Sample size determination for logistic regression revisited.

There is no consensus on the approach to compute the power and sample size with logistic regression. Some authors use the likelihood ratio test; some use the test on proportions; some suggest various approximations to handle the multivariate case. We advocate the use of the Wald test since the Z-score is routinely used for statistical significance testing of regression coefficients. The null-variance formula became popular from early studies, which contradicts modern software, which utilizes the method of maximum likelihood estimation (MLE), when the variance of the MLE is estimated at the MLE, not at the null. We derive general Wald-based power and sample size formulas for logistic regression and then apply them to binary exposure and confounder to obtain a closed-form expression. These formulas are applied to minimize the total sample size in a case-control study to achieve a given power by optimizing the ratio of controls to cases. Approximately, the optimal number of controls to cases is equal to the square root of the alternative odds ratio. Our sample size and power calculations can be carried out online at www.dartmouth.edu/ approximately eugened.

Clinical Trials as Topic↗

PubMed2007

Lysosomes and trivalent arsenic treatment in acute promyelocytic leukemia.

BACKGROUND: Cells from patients with t(15;17) acute promyelocytic leukemia (APL) express the fusion protein between the promyelocytic leukemia protein and retinoic acid receptor alpha (PML/RAR alpha). Patients with APL respond to differentiation therapy with all-trans-retinoic acid, which induces PML/RAR alpha degradation. When resistance to all-trans-retinoic acid develops, an effective treatment is arsenic trioxide (arsenite), which also induces this degradation. We investigated the mechanism of arsenite-induced PML/RAR alpha degradation. METHODS: NB4-S1 APL cells were treated with clinically relevant concentrations of arsenite. Lysosomes were visualized with a lysosome-specific dye. Lysosomal protein esterase was measured by immunoblot analysis. Lysosomal cathepsin L was detected by immunogold labeling and transmission electron microscopy, and its activity was measured in cytosolic cellular fractions. In vitro degradation assays of PML/RAR alpha in cell lysates were performed with and without protease inhibitors and assessed by immunoblot analysis. Only nonparametric two-sided statistical analyses were used. The nonparametric Wilcoxon test was used for group comparison, and the nonlinear regression technique was used for analysis of dose-response relationship as a function of arsenite concentration. RESULTS: Arsenite treatment destabilized lysosomes in APL cells. Lysosomal proteases, including cathepsin L, were released from lysosomes 5 minutes to 6 hours after arsenite treatment. PML/RAR alpha was degraded by lysate from arsenite-treated APL cells, and the degradation was inhibited by protease inhibitors. At both 6 and 24 hours, substantially fewer arsenite-treated APL cells, than untreated cells, contained cathepsin L clusters, a reflection of cathepsin L delocalization. Cells with cathepsin L clusters decreased as a function of arsenite concentration at rates of -2.03% (95% confidence interval [CI] = -4.01 to -.045; P = .045) and -2.39% (95% CI = -4.54 to -.024; P = .029) in 6- and 24-hour treatment groups, respectively, per 1.0 microM increase in arsenite concentration. Statistically significantly higher cytosolic cathepsin L activity was detected in lysates of arsenite-treated APL cells than in control lysates. For example, the mean increase in cathepsin activity at 6 hours and 1.0 microM arsenite was 26.3% (95% CI = 3.3% to 33%; P < .001), compared with untreated cells. CONCLUSIONS: In APL cells, arsenite may cause rapid destabilization of lysosomes.

Antineoplastic Agents↗

PubMed2006

In vivo EPR dosimetry to quantify exposures to clinically significant doses of ionising radiation.

As a result of terrorism, accident or war, populations potentially can be exposed to doses of ionising radiation that could cause direct clinical effects within days or weeks. There is a critical need to determine the magnitude of the exposure to individuals so that those with significant risk can have appropriate procedures initiated immediately, while those without a significant probability of acute effects can be reassured and removed from the need for further consideration in the medical/emergency system. It is extremely unlikely that adequate dosemeters will be worn by the potential victims, and it also will be unlikely that prompt and accurate dose reconstruction at the level of individuals will be possible. Therefore, there is a critical need for a method to measure the dose from radiation-induced effects that occur within the individual. In vivo EPR measurements of radiation-induced changes in the enamel of teeth is a method, perhaps the only such method, which can differentiate among doses sufficiently to classify individuals into categories for treatment with sufficient accuracy to facilitate decisions on medical treatment. In its current state, the in vivo EPR dosemeter can provide estimates of absorbed dose of +/- 0.5 Gy in the range from 1 to >10 Gy. The lower limit and the precision are expected to improve, with improvements in the resonator and the algorithm for acquiring and calculating the dose. In its current state of development, the method is already sufficient for decision-making action for individuals with regard to acute effects from exposure to ionising radiation for most applications related to terrorism, accidents or nuclear warfare.

Body Burden↗

PubMed2006

Pretreatment photosensitizer dosimetry reduces variation in tumor response.

PURPOSE: To compensate for photosensitizer uptake variation in photodynamic therapy (PDT), via control of delivered light dose through photodynamic dose calculation based on online dosimetry of photosensitizer in tissue before treatment. METHODS AND MATERIALS: Photosensitizer verteporfin was quantified via multiple fluorescence microprobe measurements immediately before treatment. To compensate individual PDT treatments, photodynamic doses were calculated on an individual animal basis, by matching the light delivered to provide an equal photosensitizer dose multiplied by light dose. This was completed for the lower quartile, median, and upper quartile of the photosensitizer distribution. PDT-induced tumor responses were evaluated by the tumor regrowth assay. RESULTS: Verteporfin uptake varied considerably among tumors and within a tumor. The coefficient of variation in the surviving fraction was found significantly decreased in groups compensated to the lower quartile (CL-PDT), the median (CM-PDT), and the upper quartile (CU-PDT) of photosensitizer distribution. The CL-PDT group was significantly less effective compared with NC-PDT (Noncompensated PDT), CM-PDT, and CU-PDT treatments. No significant difference in effectiveness was observed between NC-PDT, CM-PDT, and CU-PDT treatment groups. CONCLUSIONS: This research suggests that accurate quantification of tissue photosensitizer levels and subsequent adjustment of light dose will allow for reduced subject variation and improved treatment consistency.

Algorithms↗

PubMed2006

Arsenic exposure is associated with decreased DNA repair in vitro and in individuals exposed to drinking water arsenic.

The mechanism(s) by which arsenic exposure contributes to human cancer risk is unknown ; however, several indirect cocarcinogenesis mechanisms have been proposed. Many studies support the role of As in altering one or more DNA repair processes. In the present study we used individual-level exposure data and biologic samples to investigate the effects of As exposure on nucleotide excision repair in two study populations, focusing on the excision repair cross-complement 1 (ERCC1) component. We measured drinking water, urinary, or toenail As levels and obtained cryopreserved lymphocytes of a subset of individuals enrolled in epidemiologic studies in New Hampshire (USA) and Sonora (Mexico). Additionally, in corroborative laboratory studies, we examined the effects of As on DNA repair in a cultured human cell model. Arsenic exposure was associated with decreased expression of ERCC1 in isolated lymphocytes at the mRNA and protein levels. In addition, lymphocytes from As-exposed individuals showed higher levels of DNA damage, as measured by a comet assay, both at baseline and after a 2-acetoxyacetylaminofluorene (2-AAAF) challenge. In support of the in vivo data, As exposure decreased ERCC1 mRNA expression and enhanced levels of DNA damage after a 2-AAAF challenge in cell culture. These data provide further evidence to support the ability of As to inhibit the DNA repair machinery, which is likely to enhance the genotoxicity and mutagenicity of other directly genotoxic compounds, as part of a cocarcinogenic mechanism of action.

Adult↗

PubMed2006

Statistical comparison of color cancer cell images.

A proper analysis and comparison of medical images requires an objective criterion for image discrimination. We developed a statistical approach for color image comparison and showed how it can be applied to discriminate cancer histology images from four treatment groups. We applied a well-established multivariate analysis of variances (MANOVA) on the logit scale so that image comparison was reduced to the classic chi-square test.

Animals↗

PubMed2005

Increased oxygenation of intracranial tumors by efaproxyn (efaproxiral), an allosteric hemoglobin modifier: In vivo EPR oximetry study.

PURPOSE: To determine quantitatively the changes in oxygenation of intracranial tumors induced by efaproxiral, an allosteric hemoglobin modifier. Efaproxiral reduces hemoglobin-oxygen binding affinity, which facilitates oxygen release from hemoglobin into surrounding tissues and potentially increases the pO(2) of the tumors. METHODS AND MATERIALS: The study was performed on 10 male Fisher 344 rats with 9L intracranial tumors. Electron paramagnetic resonance (EPR) oximetry was used to measure quantitatively the changes in the pO(2) in the tumors. Lithium phthalocyanine (LiPc) crystals were implanted in the tumors and in the normal brain tissue in the opposite hemispheres. We monitored the cerebral pO(2) starting 7 to 10 days after the tumor cells were implanted. NMR imaging determined the position and size of tumor in the brain. After an initial baseline EPR measurement, efaproxiral (150 mg/kg) was injected intravenously over 15 minutes, and measurements of tumor and normal brain oxygen tension were made alternately at 10-minute intervals for the next 60 minutes; the procedure was repeated for 6 consecutive days. RESULTS: Efaproxiral significantly increased the pO(2) of both the intracranial tumors and the normal brain tissue on all days. The maximum increase was reached at 52.9 to 59.7 minutes and 54.1 to 63.2 minutes after injection, respectively. The pO(2) returned to baseline values at 106 to 126.5 minutes after treatment. The maximum tumor and normal tissue pO(2) values achieved after efaproxiral treatment from Day 1 through Day 6 ranged from 139.7 to 197.7 mm Hg and 103.0 to 135.9 mm Hg, respectively. The maximum increase in tumor tissue pO(2) values from Day 2 to Day 5 was greater than the maximum increase in normal tissue pO(2). CONCLUSION: We obtained quantitative data on the timing and extent of efaproxiral-induced changes in the pO(2) of intracerebral 9L tumors. These results illustrate a unique and useful capability of in vivo EPR oximetry to obtain repeated noninvasive measurements of tumor oxygenation over a number of days. The information on the dynamics of tumor pO(2) after efaproxiral administration illustrates the ability of efaproxiral to increase intracranial tumor oxygenation.

Aniline Compounds↗

PubMed2005

Influence analysis for linear mixed-effects models.

In this paper, we extend several regression diagnostic techniques commonly used in linear regression, such as leverage, infinitesimal influence, case deletion diagnostics, Cook's distance, and local influence to the linear mixed-effects model. In each case, the proposed new measure has a direct interpretation in terms of the effects on a parameter of interest, and collapses to the familiar linear regression measure when there are no random effects. The new measures are explicitly defined functions and do not necessitate re-estimation of the model, especially for cluster deletion diagnostics. The basis for both the cluster deletion diagnostics and Cook's distance is a generalization of Miller's simple update formula for case deletion for linear models. Pregibon's infinitesimal case deletion diagnostics is adapted to the linear mixed-effects model. A simple compact matrix formula is derived to assess the local influence of the fixed-effects regression coefficients. Finally, a link between the local influence approach and Cook's distance is established. These influence measures are applied to an analysis of 5-year Medicare reimbursements to colon cancer patients to identify the most influential observations and their effects on the fixed-effects coefficients.

Cluster Analysis↗

PubMed2005

Simultaneous measurement of rat brain cortex PtO2 using EPR oximetry and a fluorescence fiber-optic sensor during normoxia and hyperoxia.

Electron paramagnetic resonance (EPR) oximetry is a promising, relatively non-invasive method of monitoring tissue partial pressure of oxygen (PtO(2)) that has proven useful in following changes in PtO(2) under various physiologic and pathophysiologic conditions. Optimal utilization of the method will be facilitated by systematic comparisons with other available methods. Here, we report on the absolute values and changes of rat brain PtO(2) using EPR oximetry and the OxyLite, an oxygen monitor based on fluorescence quenching, at adjacent locations in the same brain. EPR oximetry utilizes an implanted oxygen-sensitive material and reports tissue PtO(2) at the surface of the material. OxyLite measures PtO(2) using the fluorescence lifetime of a chromophore fixed to the tip of an optical fiber that is inserted into tissue. Measurements were made at a depth of 2-3 mm into the cortex during normoxia and during breathing of carbogen (95% O(2):5% CO(2)) followed by a return to normoxia. We conclude that in this study (1) PtO(2) values reported by the two methods are similar but not exactly the same, (2) both methods can record a baseline and rapid changes in PtO(2), (3) changes in PtO(2) induced by increasing FiO(2) from 0.26 to 0.95 (carbogen) were similar by the two methods and (4) in some rats breathing carbogen, absolute values of PtO(2) were above the sensitive range of the OxyLite method.

Animals↗

PubMed2005

Effect of tumor host microenvironment on photodynamic therapy in a rat prostate tumor model.

PURPOSE: Tumor host microenvironment plays an important role in tumor growth, metastasis, and response to cancer therapy. In this study, the influence of tumor host environment on tumor pathophysiology, photosensitizer distribution, and photodynamic therapy (PDT) treatment effect was examined in the metastatic at lymph node and lung (MatLyLu) rat prostate tumor. EXPERIMENTAL DESIGN: MatLyLu tumors implanted in different host environment [i.e., orthotopically (in the prostate) or s.c.] were compared for difference in vessel density, average vessel size, vascular permeability, tumor vascular endothelial growth factor production, and tumor oxygenation. Uptake of photosensitizer verteporfin in tumors in both sites was determined by fluorescence microscopy. To compare tumor response to PDT, both orthotopic and s.c. MatLyLu tumors were given the same doses of verteporfin and laser light treatment, and PDT-induced tumor necrotic area was measured histologically. RESULTS: Orthotopic MatLyLu tumors were found to grow faster, have higher vessel density and more permeable vasculature, have higher vascular endothelial growth factor protein levels, and have lower tumor hypoxic fraction than the s.c. tumors. Uptake of photosensitizer verteporfin in the orthotopic tumor was higher than in the s.c. tumors at 15 minutes after injection (1 mg/kg, i.v.), and became similar at 3 hours after injection. For the vascular targeting PDT treatment (0.25 mg/kg verteporfin, 50 J/cm(2) at 50 mW/cm(2), 15 minutes drug-light interval), there was no significant difference in PDT-induced tumor necrotic area between the orthotopic and s.c. tumors, with 85% to 90% necrosis in both types of tumors. However, tumor necrosis induced by the cellular targeting PDT (1 mg/kg verteporfin, 50 J/cm(2) at 50 mW/cm(2), 3 hours drug-light interval) was significantly different in the orthotopic (64%) versus the s.c. (29%) tumors. CONCLUSIONS: Tumor host environment can significantly affect photosensitizer verteporfin distribution and PDT treatment effect. Verteporfin-PDT regimen targeting tumor cells is more sensitive to such influence than the vascular targeting PDT. Our study showed the importance of tumor host environment in determining tumor physiologic properties and tumor response to PDT. To obtain clinically relevant information, orthotopic tumor model should be used in the experimental studies.

Animals↗

PubMed2005

Measurement error and confidence intervals for ROC curves.

Measurement error in a continuous test variable may bias estimates of the summary properties of receiver operating characteristics (ROC) curves. Typically, unbiased measurement error will reduce the diagnostic potential of a continuous test variable. This paper explores the effects of possibly heterogenous measurement error on estimated ROC curves for binormal test variables. Corrected estimators for specific points on the curve are derived under the assumption of known or estimated measurement variances for individual test results. These estimators and associated confidence intervals do not depend on normal assumptions for the distribution of the measurement error and are shown to be approximately unbiased for moderate size samples in a simulation study. An application from a study of emerging imaging modalities in breast cancer is used to demonstrate the new techniques.

Computer Simulation↗

PubMed2005

Measurements of clinically significant doses of ionizing radiation using non-invasive in vivo EPR spectroscopy of teeth in situ.

There are plausible circumstances in which populations potentially have been exposed to doses of ionizing radiation that could cause direct clinical effects within days or weeks, but there is no clear knowledge as to the magnitude of the exposure to individuals. In vivo EPR is a method, perhaps the only such method that can differentiate among doses sufficiently to classify individuals into categories for treatment with sufficient accuracy to facilitate decisions on medical treatment. Individuals with significant risk then can have appropriate procedures initiated immediately, while those without a significant probability of acute effects could be reassured and removed from the need for further medical treatment. In its current state, the in vivo EPR dosimeter can provide estimates of absorbed dose of +/-25 cGy in the range of 100-->1000 cGy. This is expected to improve, with improvements in the resonator, the algorithm for calculating dose, and the uniformity of the magnetic field. In its current state of development, it probably is sufficient for most applications related to terrorism or nuclear warfare, for decision-making for action for individuals in regard to acute effects from exposure to ionizing radiation.

Algorithms↗

PubMed2005

Cerebral tissue oxygenation in reversible focal ischemia in rats: multi-site EPR oximetry measurements.

Multi-site electron paramagnetic resonance (EPR) oximetry was used in vivo to measure the partial pressure of oxygen (pO2) in reversible focal ischemia in rats. The cerebral tissue pO2 was measured simultaneously and continuously at two sites on the ischemic side and one on the normal side of the brain in the same animal prior to and at several time points after ischemia and reperfusion. The O2 at the three different sites in brain was stable over 30 min of baseline measurements. During the first 120 min of ischemia, statistically significant decreases in brain pO2 from baseline were consistently observed in the ischemic core and perifocal area. The mean values varied during the 120 min of ischemia. Reperfusion resulted in an immediate increase in PO2, but there were no significant differences between the sites over time. The result of this study seems promising for the study of ischemia and reperfusion. It appears that the technique can provide information on the PO2 under the experimental conditions needed for such a study. The levels of PO2 that occurred in these experiments are readily resolvable by multi-site EPR oximetry. In addition, the ability simultaneously to measure the pO2 in several sites provides important additional information that should help to differentiate between changes in the PO2 due toglobal or local mechanisms.

Animals↗

PubMed2005

On optimal current patterns for electrical impedance tomography.

We develop a statistical criterion for optimal patterns in planar circular electrical impedance tomography. These patterns minimize the total variance of the estimation for the resistance or conductance matrix. It is shown that trigonometric patterns (Isaacson, 1986), originally derived from the concept of distinguishability, are a special case of our optimal statistical patterns. New optimal random patterns are introduced. Recovering the electrical properties of the measured body is greatly simplified when optimal patterns are used. The Neumann-to-Dirichlet map and the optimal patterns are derived for a homogeneous medium with an arbitrary distribution of the electrodes on the periphery. As a special case, optimal patterns are developed for a practical EIT system with a finite number of electrodes. For a general nonhomogeneous medium, with no a priori restriction, the optimal patterns for the resistance and conductance matrix are the same. However, for a homogeneous medium, the best current pattern is the worst voltage pattern and vice versa. We study the effect of the number and the width of the electrodes on the estimate of resistivity and conductivity in a homogeneous medium. We confirm experimentally that the optimal patterns produce minimum conductivity variance in a homogeneous medium. Our statistical model is able to discriminate between a homogenous agar phantom and one with a 2 mm air hole with error probability (p-value) 1/1000.

Algorithms↗

PubMed2004

Effect of RSR13, an allosteric hemoglobin modifier, on oxygenation in murine tumors: an in vivo electron paramagnetic resonance oximetry and bold MRI study.

PURPOSE: RSR13, an allosteric modifier of hemoglobin, reduces hemoglobin-oxygen binding affinity facilitating oxygen release from hemoglobin, resulting in increases in tissue pO(2). The purpose of this study was noninvasively to monitor the time course and effect of RSR13 on tumor oxygenation, directly using in vivo electron paramagnetic resonance (EPR oximetry), and indirectly using blood oxygen level dependent magnetic resonance imaging (BOLD MRI). METHODS AND MATERIALS: The study was performed in transplanted radiation-induced fibrosarcoma tumors (RIF-1) in 18 female C3H/HEJ mice, which had two lithium phthalocyanine (LiPc) deposits implanted in the tumor when the tumors reached about 200-600 mm(3). Baseline EPR measurements were made daily for 3 days. Then, for 6 consecutive days and after an initial baseline EPR measurement, RSR13 (150 mg/kg) or vehicle (same volume) was injected intraperitoneally, and measurements of intratumoral oxygen were made at 10-min intervals for the next 60 min. In each mouse, every third day, instead of EPR oximetry, BOLD MRI measurements were made for 60 min after administration of the RSR13. RESULTS: Based on EPR measurements, RSR13 produced statistically significant temporal increases in tumor pO(2) over the 60-min time course, which reached a maximum at 35-43 min postdose. The average time required to return to the baseline pO(2) was 70-85 min. The maximum increase in tumor tissue pO(2) values after RSR13 treatment from Day 1 to Day 5 (8.3-12.4 mm Hg) was greater than the maximum tumor tissue pO(2) value for Day 6 (4.7 mm Hg, p < 0.01). The maximum increase in pO(2) occurred on Day 2 (12.4 mm Hg) after RSR13 treatment. There was little change in R(2)*, indicating that the RSR13 had minimal detectable effects on total deoxyhemoglobin and hemoglobin-oxygen saturation. CONCLUSION: The extent of the increase in tumor pO(2) achieved by RSR13 would be expected to lead to a significant increase in the effectiveness of tumor radiotherapy. The lack of a change in the BOLD MRI signal suggests that the tumor physiology was largely unchanged by RSR13. These results illustrate a unique and useful capability of in vivo EPR oximetry and BOLD MRI to obtain repeated measurements of tumor oxygenation and physiology. The dynamics of tumor pO(2) after RSR13 administration may be useful for the design of clinical protocols using allosteric hemoglobin effectors.

Aniline Compounds↗

PubMed2004

Incidence of transitional cell carcinoma of the bladder and arsenic exposure in New Hampshire.

OBJECTIVE: Arsenic is a known bladder carcinogen and populations exposed to high arsenic levels in their water supply have reported elevated bladder cancer mortality and incidence rates. To examine the effects of lower levels of arsenic exposure on bladder cancer incidence, we conducted a case-control study in New Hampshire, USA where levels above 10 micro/l are commonly found in private wells. METHODS: We studied 383 cases of transitional cell carcinoma of the bladder cancer, newly diagnosed between July 1, 1994 and June 30, 1998 and 641 general population controls. Individual exposure to arsenic was determined in toenail clippings using instrumental neutron activation analysis. RESULTS: Among smokers, an elevated odds ratio (OR) for bladder cancer was observed for the uppermost category of arsenic (OR: 2.17, 95% CI: 0.92-5.11 for greater than 0.330 mcg/g compared to less than 0.06 micro/g). Among never smokers, there was no association between arsenic and bladder cancer risk. CONCLUSIONS: These, and other data, suggest that ingestion of low to moderate arsenic levels may affect bladder cancer incidence, and that cigarette smoking may act as a co-carcinogen.

Adult↗

PubMed2004

pO2 and regional blood flow in a rabbit model of limb ischemia.

Oxygen tension (pO2) in muscles and regional blood flow were measured in a rabbit model of limb ischemia. pO2 was measured repetitively by EPR oximetry with EMS char in four different muscle groups in the same animals. Blood flow in the same muscles at several time points was measured using microspheres. A linear mixed effects model was developed to analyze the data on pO2 and blood flow. The results suggest that while under normal conditions pO2 in muscles does not depend significantly on blood flow, immediately after arterial occlusion pO2 correlates linearly with blood flow. Within two weeks of occlusion the pO2 is recovered to 45% of baseline. This study demonstrates, for the first time, the applicability of EPR oximetry in animals larger than rodents.

Algorithms↗

PubMed2004

Comparison of EPR oximetry and Eppendorf polarographic electrode assessments of rat brain PtO2.

EPR oximetry is a promising, relatively non-invasive method for monitoring the partial pressure of oxygen in tissue (PtO2) that has proved useful in following changes under various physiologic and pathophysiologic conditions. Optimal utilization of the method will be facilitated by systematic comparisons with other available methods. Here we report on the absolute values of rat brain PtO2 using EPR and the more widely used Eppendorf polarographic microelectrode system in the same brain. EPR used an L-band (1.2 GHz) spectrometer and implanted lithium phthalocyanine (LiPc) as the oxygen-sensitive paramagnetic material. Eppendorf measurements were made by a needle probe moved vertically through the cortex at 0.5 mm intervals in three tracks including one adjacent to the location of the LiPc. Several conclusions were drawn, including, (1) the average PtO2 measured by the two methods was similar but EPR reported a significantly higher average PtO2, (2) there was poor correlation between the values in the same animal on the same side of the brain, (3) the Eppendorf reported a larger range of values and (4) the heterogeneity of oxygen levels in the brain and the areas sampled by the two methods provide an adequate explanation for the observed differences.

Animals↗