[Research progress on health hazards and occupational protection of prolymethylmethacrylate bone cement in the operating room].

There are many occupational risk factors in operating room work. Polymethyl methacrylate (PMMA) , as a kind of common bone filling and repairing material, is widely used in the fixation of artificial joints, oral braces and orthopedic prosthesis. However, PMMA will release methyl methacrylate (MMA) monomer when it is implanted into human tissues and polymerized to harden, which is toxic to the body. This paper analyzes harmful factors in using PMMA bone cement, and then explores corresponding occupational protection knowledge, in order to reduce the occurrence of occupational hazards related to PMMA bone cement and enhance the self-protection ability of the operation room medical staff.

[Effect of ketogenic diet on obesity asthma].

Recently investigators pay more attention to the relationship between obesity and bronchial asthma (asthma).Obesity is increasingly recognized as a possible risk factor for childhood asthma, and 70% of patients with difficult-to-treat asthma are overweight or obese. In recent years, ketogenic diet, as one of the natural therapies, has been shown to have positive effects on weight loss process; and recent studies showed that ketogenic diet reduced airway inflammation in asthma. This review summarized the mechanisms of associations between obesity and asthma, and described the potential mechanisms of ketogenic diet affecting obese asthma, such as controlling body weight, reducing inflammatory response, regulating intestinal flora and modifying epigenetics, to provide new ideas for the prevention and treatment of obesity asthma.

Correlation between Abdominal Wall Subcutaneous Fat Thickness and Heart Weight in Southern Chinese Population.

ABSTRACT: Objective To study the correlation between the abdominal wall subcutaneous fat thickness and heart weight, so as to provide reference for prediction methods of normal range of heart weight that is suitable for autopsy in China. Methods The forensic pathology autopsy cases accepted by Center for Medicolegal Expertise of Sun Yat-sen University from 1998 to 2017 were collected. Then the exclusion criteria were determined, and according to them the total case group was selected, and the 6 disease groups and the normal group were further selected from the total case group. The rank sum test was used to compare the heart weight of the normal group and the disease groups to determine the influence of diseases on heart weight. Then the Spearman rank correlation analysis of abdominal wall subcutaneous fat thickness and heart weight in different genders and different ages in the total case group and the normal group was conducted to get the correlation coefficient （rs）. Results In the total case group, correlation between abdominal wall subcutaneous fat thickness and heart weight was shown in males of all ages （P<0.05）; while in females, the correlation had no statistical significance （P>0.05） in 15-<20 age and 50-<60 age, but was statistically significant （P<0.05） in other age groups. For the males in the normal group, rs was respectively 0.411, 0.541 and 0.683 in the 15-<40 age, the 40-<60 age, and the ≥60 age. For the females, rs was respectively 0.249 and 0.317 in the 15-<40 age and the 40-<60 age. The correlation in the ≥60 age had no statistical significance（P>0.05）. Conclusion In the general population and the normal population, abdominal wall subcutaneous fat thickness is correlated with the heart weight of males. It is of significance to include the abdominal wall subcutaneous fat thickness in the prediction of normal range of heart weight for males in China.

Application Prospects of Virtual Autopsy in Forensic Pathological Investigations on COVID-19.

ABSTRACT: Autopsy of patients who died of infectious diseases is of significance for public health management. Few forensic autopsies have been performed since the outbreak of the corona virus disease 2019 （COVID-19） due to some limitations, thus forensic pathological examination failed to contribute to the prevention and treatment of infectious diseases. Virtual autopsy has unique advantages in the forensic examination of patients who died of infectious diseases. Accumulated virtual autopsy image data are of great value to the study of the pathological mechanism and diagnosis of COVID-19. This article reviews the relationship between imaging changes and pathology of the COVID-19 as well as the application of virtual autopsy in autopsy of patients who died of infectious diseases, in order to provide reference for performing virtual autopsy in the outbreak of COVID-19.

Rotation control of a variable-capacitance electrostatic motor for space equivalence principle tests with rotating masses.

Variable-capacitance electrostatic motors are ideal for driving the test mass in ultra-low-noise electrostatic accelerometers. Such devices are essential for testing the new equivalence principle (NEP) with rotating extended masses. However, as the air-film damping is greatly reduced by placing the sensor core assembly in a high-vacuum housing, this synchronous motor may easily fall out of step and suffer spin-up failures with traditional open-loop excitation. In this study, a synchronous electronic phase commutation scheme is proposed by sensing the three-phase position change of the rotor poles and activating the stator electrodes in careful correlation with the instantaneous rotor position. Experiments on a ground-test NEP instrument prototype show that the proposed closed-loop excitation scheme can spin-up the rotor synchronously and maintain stable constant-speed operation of this macroscale variable capacitance motor operated in a high-vacuum environment. This rotation control method is also applicable to the synchronous operation of micromachined variable-capacitance electrostatic motors.

Modeling and compensation of cross-axis coupling in an electrostatic accelerometer for testing the equivalence principle.

Electrostatic accelerometers have extremely high sensitivity and are ideal scientific instruments for measuring very weak acceleration. In particular, a single-sensitive-axis electrostatic accelerometer can be used for testing the equivalence principle in space. Sensitive-axis capacitances formed by axial electrodes and a cylindrical proof mass vary with the axial motion of the mass and are also affected by radial motion, which results in cross-axis coupling disturbances. A quantitative model is built to analyze the cross-axis coupling effect on the sensitive axis from the radial suspension loop, including a nonlinear model for large radial motion and a linear model for small radial motion. Frequency response simulation shows that the cross-axis coupling effect for a small signal case arises mostly in the high-frequency range. Experiments are carried out with a ground-based electrostatic accelerometer made of a single, non-rotating test cylinder, and in this case, the experimental results are utilized to verify the mathematical model. Cross-axis coupling for small signal perturbations is virtually removed if the equilibrium position of the proof mass is calibrated to the null position of the sensor cage. In addition, data post-processing can further attenuate the cross-axis coupling disturbances when dealing with large radial motion. The cross-axis coupling disturbances on both the position and the acceleration measurement signals in the sensitive axis are mostly removed in ground-based experiments. The proposed model and compensation can be extended to space equivalence principle instruments and other electrostatic accelerometers with a cylindrical proof mass.

Microgravity Level Measurement of the Beijing Drop Tower Using a Sensitive Accelerometer.

Drop tower is the most common ground-based facility to provide microgravity environment and widely used in many science experiments. A differential space accelerometer has been proposed to test the spin-gravity interaction between rotating extended bodies onboard a drag-free satellite. In order to assist design and test of this inertial sensor in a series of ground- based pre-flight experiments, it is very important to know accurately the residual acceleration of drop towers. In this report, a sensitive instrument for this purpose was built with a high-performance servo quartz accelerometer, and the dedicated interface electronics design providing small full-scale range and high sensitivity, up to 136.8 V/g0. The residual acceleration at the Beijing drop tower was measured using two different drop capsules. The experimental result shows that the microgravity level of the free-falling double capsule is better than 2 × 10(-4)g0 (Earth's gravity). The measured data in this report provides critical microgravity information for design of the following ground experiments.

Detection and prevalence of pathogenic Yersinia enterocolitica in refrigerated and frozen dairy products by duplex PCR and dot hybridization targeting the virF and ail genes.

Pathogenic Yersinia enterocolitica is involved in yersiniosis through expression of chromosome-borne or plasmid-borne virulence factors. Yersinia enterocolitica is a cold-tolerant pathogen frequently isolated from refrigerated or frozen foods. However, little attention has been focused on the prevalence of pathogenic Y. enterocolitica in refrigerated or frozen dairy samples in China. In this study, we developed a new duplex PCR targeting the plasmid-borne virF gene and chromosome-borne ail gene for detection of pathogenic Y. enterocolitica isolates. We established a detection limit for the duplex PCR of 6.5 × 10(2)cfu/mL in artificially contaminated dairy samples. In addition, the duplex PCR could detect directly 4.5 to 5.7 cfu of Y. enterocolitica in 5 mL of brain heart infusion broth after 6 h of enrichment at 28 °C. A newly developed dot hybridization assay further confirmed specificity of the duplex PCR for detection of virulent Y. enterocolitica. Furthermore, 13 Y. enterocolitica and 5 pathogenic strains, from 88 commercial frozen or refrigerated dairy products, were detected successfully by the China National Standard method (GB/T4789.8-2008) and the duplex PCR, respectively. Finally, biotypes and serotypes of pathogenic Y. enterocolitica strains were further characterized. The duplex PCR developed here is reliable for large-scale screening, routine monitoring, and risk assessment of pathogenic Y. enterocolitica in refrigerated or frozen dairy products.

Screening of biomarkers for lung cancer with gene expression profiling data.

OBJECTIVES: Lung cancer is one of the most common malignant tumors, but the etiology is not yet clear. Our study aims to deepen the understandings about the mechanisms of lung cancer via screening relevant key genes and functional pathways. MATERIALS AND METHODS: Microarray data set was collected and differentially expressed genes (DEGs) were selected out. KEGG pathway analysis and Gene Ontology (GO) enrichment analysis were performed for the DEGs. Interaction networks were constructed for the lung cancer-related DEGs with information from Human Protein Reference Database (HRPD) to screen out potential biomarkers. RESULTS: Functional annotation revealed that cell cycle, DNA replication, immune system, and signal molecules and interactions were significantly over-represented in all the DEGs, suggesting their close involvement in the development of lung cancer. 40 genes with high degree, betweenness and clustering coefficient were identified from the interaction network. 26 out of them are known cancer genes according to the database F-census. Besides, 4 biomarkers were revealed through analyzing their interactions with oncogenes. CONCLUSIONS: Our study not only advances the understandings about the molecular mechanisms of lung cancer, but also provides several potential biomarkers for clinical use.

Effects of ozone on membrane permeability and ultrastructure in Pseudomonas aeruginosa.

AIMS: To examine the mechanism of ozone-induced damage to cytoplasmic membrane and cell ultrastructure of Pseudomonas aeruginosa ATCC27853. METHODS AND RESULTS: Cell suspensions of Ps. aeruginosa ATCC27853 were treated with ozonated water. The leakages of cellular potassium (K⁺), magnesium (Mg²âº) and adenosine triphosphate (ATP), determined by inductively coupled plasma/mass spectrometry (ICP/MS) and a commercial bioluminescence assay kit, were to assess ozone-induced damage to the cytoplasmic membrane. Maximum leakages of K⁺ and Mg²âº were attained, respectively, at 0·53 mg l⁻¹ ozone after 0·5 and 2 min with > 99% inactivation of culturable bacteria, while that of ATP was achieved at 0·67 mg l⁻¹ ozone after 1 min. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) revealed that treated cells retained intact shapes and cytoplasm agglutinations and vacuoles occurred. CONCLUSIONS: Ozone inactivates Ps. aeruginosa ATCC27853 by the combined results of increased cytoplasmic membrane permeability and cytoplasm coagulation, rather than by severe membrane disruption and cell lysis. SIGNIFICANCE AND IMPACT OF THE STUDY: Pseudomonas aeruginosa is a common water-related pathogen. These insights into the leakage of cytoplasmic components and ultrastructural changes provide evidence for the mechanisms of ozone-mediated inactivation.

Effects of L-malate on mitochondrial oxidoreductases in liver of aged rats.

Accumulation of oxidative damage has been implicated to be a major causative factor in the decline in physiological functions that occur during the aging process. The mitochondrial respiratory chain is a powerful source of reactive oxygen species (ROS), considered as the pathogenic agent of many diseases and aging. L-malate, a tricarboxylic acid cycle intermediate, plays an important role in transporting NADH from cytosol to mitochondria for energy production. Previous studies in our laboratory reported L-malate as a free radical scavenger in aged rats. In the present study we focused on the effect of L-malate on the activities of electron transport chain in young and aged rats. We found that mitochondrial membrane potential (MMP) and the activities of succinate dehydrogenase, NADH-cytochrome c oxidoreductase and cytochrome c oxidase in liver of aged rats were significantly decreased when compared to young control rats. Supplementation of L-malate to aged rats for 30 days slightly increased MMP and improved the activities of NADH-dehydrogenase, NADH-cytochrome c oxidoreductase and cytochrome c oxidase in liver of aged rats when compared with aged control rats. In young rats, L-malate administration increased only the activity of NADH-dehydrogenase. Our result suggested that L-malate could improve the activities of electron transport chain enzymes in aged rats.

Plasma membrane damage to Candida albicans caused by chlorine dioxide (ClO2).

AIM: To investigate the plasma membrane damage of chlorine dioxide (ClO(2)) to Candida albicans ATCC10231 at or below the minimal fungicidal concentration (MFC). METHODS AND RESULTS: ClO(2) at MFC or below was adopted to treat the cell suspensions of C. albicans ATCC10231. Using transmission electron microscopy, no visible physiological alteration of cell shape and plasma membrane occurred. Potassium (K(+)) leakages were significant; likewise, it showed time- and dose-dependent increases. However, adenosine triphosphate (ATP) leakages were very slight. Research shows that when 99% of the cells were inactivated, the leakage was measured at 0.04% of total ATP. Compared with the mortality-specific fluorescent dye of DiBAC(4)(3), majority of the inactivated cells were poorly stained by propidium iodide, another mortality-specific fluorescent dye which can be traced by flow cytometry. CONCLUSION: At or below MFC, ClO(2) damages the plasma membranes of C. albicans mainly by permeabilization, rather than by the disruption of their integrity. K(+) leakage and the concomitant depolarization of the cell membrane are some of the critical events. SIGNIFICANCE AND IMPACT OF THE STUDY: These insights into membrane damages are helpful in understanding the action mode of ClO(2).

L-malate reverses oxidative stress and antioxidative defenses in liver and heart of aged rats.

The intracellular levels of antioxidant and free radical scavenging enzymes are gradually altered during the aging process. An age-dependent increase of oxidative stress occurring throughout the lifetime is hypothesized to be the major cause of aging. The current study examined the effects of L-malate on oxidative stress and antioxidative defenses in the liver and heart of aged rats. Sprague-Dawley male rats were randomly divided into four groups, each group consisting of 6 animals. Group Ia and Group IIa were young and aged control rats. Group Ib and Group IIb were young and aged rats treated with L-malate (210 mg/kg body weight per day). L-malate was orally administrated via intragastric canula for 30 days, then the rats were sacrificed and the liver and heart were removed to determine the oxidant production, lipid peroxidation and antioxidative defenses of young and aged rats. Dietary L-malate reduced the accumulation of reactive oxygen species (ROS) and significantly decreased the level of lipid peroxidation in the liver and heart of the aged rats. Accordingly, L-malate was found to enhance the antioxidative defense system with an increased activity of antioxidant enzymes, such as superoxide dismutase (SOD) and glutathione peroxidase (GPx) and increased glutathione (GSH) levels in the liver of aged rats, a phenomenon not observed in the heart of aged rats. Our data indicate that oxidative stress was reversed and the antioxidative defense system was strengthened by dietary supplementation with L-malate.

Effects of L-malate on physical stamina and activities of enzymes related to the malate-aspartate shuttle in liver of mice.

L-malate, a tricarboxylic acid cycle (TCA) intermediate, plays an important role in transporting NADH from cytosol to mitochondria for energy production and may be involved in the beneficial effects of improving physical stamina. In the present study, we investigated the effects of L-malate on the performance of forced swimming time and blood biochemical parameters related to fatigue - blood urea nitrogen (BUN), glucose (Glc), creatine kinase (CK),total protein (TP) and lactic acid (LA). To investigate the effects of L-malate on the malate-aspartate shuttle and energy metabolism in mice, the activities of enzymes related to the malate-aspartate shuttle were measured. L-malate was orally administered to mice continuously for 30 days using a feeding atraumatic needle. The swimming time was increased by 26.1 % and 28.5 %, respectively, in the 0.210 g/kg and 0.630 g/kg L-malate-treated group compared with the control group. There were no differences in the concentrations of Glc, BUN and TP between the L-malate-treated groups and the control groups. However, the levels of CK were significantly decreased in the L-malate-treated groups. The results predict a potential benefit of L-malate for improving physical stamina and minimizing muscle damage during swimming exercise. The activities of cytosolic and mitochondrial malate dehydrogenase were significantly elevated in the L-malate-treated group compared with the control group. These enzymatic activities may be useful indicators for evaluating changes affecting the malate-aspartate shuttle and energy metabolism in the liver of mice.

Conjugation with (111)In-DTPA-poly(ethylene glycol) improves imaging of anti-EGF receptor antibody C225.

UNLABELLED: Significant liver uptake often limits the clinical application of radiolabeled antibodies in radioimmunodetection. The purpose of this study was to evaluate the gamma-imaging properties of an antiepidermal growth factor receptor (EGFR) antibody, C225, conjugated with heterofunctional poly(ethylene glycol) (PEG) with 1 terminus of the polymer attached to a radiometal chelator, diethylenetriaminepentaacetic acid (DTPA). METHODS: Two preparations of PEG-modified C225, one with 20% and the other with 60% amine substitution, were labeled with (111)In. The conjugates, (111)In-DTPA-PEG-C225, were injected intravenously into nude mice with EGFR-positive A431 tumors. For comparison, C225 directly labeled with (111)In was also injected. In a competitive study, mice with A431 tumors were pretreated intravenously with 100-fold excess of native C225, followed by an injection of (111)In-DTPA-PEG-C225 30 min or 20 h later. In addition, (111)In-DTPA-PEG-C225 was injected into mice with EGFR-positive MDA-MB-468 tumors and EGFR-negative MDA-MB-435 tumors. Images were acquired at 5 min and at 2, 6, 24, and 48 h after injection of the radiotracers. Regions of interest (ROIs) were drawn on the computer images around the whole body, liver, muscle, and tumor. The counts per pixel in the tumor and normal tissues were calculated. At 48 h, the mice were killed and dissected. Blood, liver, muscle, and tumor samples were removed and the radioactivity of each sample was measured. RESULTS: In A431 tumor xenografts, the tumor uptake of C225 modified with PEG was not significantly different than the uptake of unmodified (111)In-DTPA-C225. Uptake in the liver, however, was reduced by 38%-45%, and the reduction increased with increasing degree of PEG substitution. Tumors of A431 and MDA-MB-468 xenografts were clearly visualized with (111)In-DTPA-PEG-C225, whereas tumors of the MDA-MB-435 xenograft, which expresses low levels of EGFR, were not as readily visible. The tumor-to-blood ratios of (111)In-DTPA-PEG-C225 in A431 and MDA-MB-468 xenografts were about 3 fold higher than in MDA-MB-435 xenografts. Blocking EGFR by pretreatment with native C225 significantly reduced the uptake of (111)In-DTPA-PEG-C225 in the liver. The tumor-to-blood ratios in mice with A431 tumors were decreased 2.5-2.7 fold after pretreatment with a large excess of C225. Similar results were obtained with MDA-MB-468 tumor xenografts. In contrast, the tumor-to-blood ratios in mice with MDA-MB-435 tumor xenografts were not significantly different in C225-pretreated mice than in nonpretreated mice. CONCLUSION: These findings indicate that (111)In-DTPA-PEG-C225 selectively localized to the tumors expressing high levels of EGFR. PEG-modification of C225 significantly reduced its liver uptake, resulting in improved visualization of EGFR-positive tumors. Using PEG as a linker between the monoclonal antibody and metal chelator is a useful strategy to optimize the imaging characteristics of antibody-based scintigraphic agents.

Poly(ethylene glycol)-conjugated anti-EGF receptor antibody C225 with radiometal chelator attached to the termini of polymer chains.

Several biological barriers, including significant liver uptake, limit the clinical application of radiolabeled antibodies in radioimmunoscintigraphy. Here, a general approach is described for radiolabeling of monoclonal antibodies conjugated with poly(ethylene glycol) (PEG). This strategy is demonstrated with C225, a monoclonal antibody directed against epidermal growth factor (EGF) receptor. We synthesized a heterofunctional PEG with one end attached to a radiometal chelator, diethylenetriaminepentaacetic acid (DTPA), and the other end to a protected thiol group, S-acetylthioacetate. After a deprotection step, the resulting DTPA-PEG-SH was conjugated to maleimide-activated C225 to yield DTPA-PEG-C225 conjugate. Characterization of DTPA-PEG-C225 with immunoprecipitation and Western blot analysis revealed that the conjugate was biologically active in binding to the EGF receptor in A431 cells. Competitive EGF receptor binding assay in MDA-MB-468 cells showed that DTPA-PEG-C225, with up to 60% of the amino groups in C225 substituted, retained 66% of C225's binding affinity. Moreover, DTPA-PEG-C225 with increasing degrees of NH(2) substitution from 20% to 70% retained the activity of C225 to induce apoptosis in DiFi cells. More importantly, DTPA-PEG-C225 demonstrated less nonspecific interaction than DTPA-C225. Pharmacokinetic analysis using (111)In-labeled compounds revealed narrower steady-state distribution of (111)In-DTPA-PEG-C225 than (111)In-DTPA-C225, probably due to reduced nonspecific binding of PEG-modified antibody to tissues. The terminal half-life (t(1/2,)(gamma)) of (111)In-DTPA-PEG-C225, 21.1 h, was shorter than that of (111)In-DTPA-C225, 52.9 h. These data suggest that (111)In-DTPA-PEG-C225 may provide better imaging characteristics than (111)In-DTPA-C225, and that using PEG as a linker between the monoclonal antibody and DTPA may be a promising strategy in optimizing the imaging characteristics of immunoscintigraphic agents.

Effectiveness of water soluble poly(L-glutamic acid)-camptothecin conjugate against resistant human lung cancer xenografted in nude mice.

A camptothecin (CPT) formulation that can be easily administered, is less toxic, and has greater antitumor effect is needed. In this study, a water-soluble CPT derivative was obtained by direct coupling of CPT to poly(L-glutamic acid) (PG) through the C20(S)-hydroxyl group. CPT was released from the resulting conjugate, PG-CPT, in phosphate-buffered saline with a zero-order kinetics in the initial 50 days. The release rates were 0.623% per day, 1.081% per day, and 1.396% per day at pH 5.3, 7.4, and 9.0, respectively. In vitro, PG-CPT was less potent in inhibiting cell growth than was free CPT in all human tumor cell lines tested. However, PG-CPT showed better antitumor activity and tolerability than did CPT in vivo. When H322 human lung tumor cells were inoculated subcutaneously in nude mice, PG-CPT delayed the growth of these well-established tumors with an absolute growth delay of 32 days when given as 4 doses with 4-day intervals between injections at an equivalent CPT dose of 40 mg/kg. When H322 cells were inoculated intratracheally in nude mice, 5 doses of intravenous injection of PG-CPT at an equivalent CPT dose of 10 mg/kg on days 4, 8, 12, 16, and 20 after inoculation significantly prolonged the median survival of treated mice, averaging 1.8-fold that of untreated mice (p=0.01). Increasing the dose of PG-CPT to an equivalent CPT dose of 40 mg/kg per injection administered in 4 doses on days 4, 8, 12, and 16 prolonged the median survival of treated mice by 4-fold (p=0.0008). Significantly, mice with intratracheally inoculated H322 tumors were resistant to both CPT and cisplatin treatments. These studies demonstrated that PG may be used as an effective solubilizing carrier for CPT and that PG-CPT may have potential application in the treatment of lung cancer.

Biodistribution of paclitaxel and poly(L-glutamic acid)-paclitaxel conjugate in mice with ovarian OCa-1 tumor.

PURPOSE: Poly(L-glutamic acid)-paclitaxel (PG-TXL) is a water-soluble paclitaxel (TXL) conjugate made by conjugating TXL to poly(L-glutamic acid) via ester bonds. In preclinical studies, PG-TXL has shown significant antitumor activity against a variety of solid tumors. To elucidate the relationship between tissue distribution and antitumor efficacy of PG-TXL, we studied and compared the biodistribution of PG-TXL and TXL. METHODS: Female C3Hf/Kam mice bearing syngeneic ovarian OCa-1 tumors were injected with either [3H]TXL or PG-[3H]TXL at an equivalent TXL dose of 20 mg/kg. Mice were killed at various times after drug injection, and samples of blood, spleen, liver, kidney, lung, heart, muscle, brain, fat, and tumor were removed and the radioactivity counted. In addition, concentrations of free [3H]TXL released from PG-[3H]TXL in the spleen, liver, kidney, and tumor were analyzed by using high-performance liquid chromatography (HPLC). Whole-body autoradiographs of mice killed 1 day and 6 days after administration of PG-[3H]TXL were obtained to study the intratumoral distribution of PG-TXL. RESULTS: When [3H]TXL was conjugated to polymer, the biodistribution pattern of PG-[3H]TXL differed from that of [3H]TXL. Based on area under the tissue concentration-time curve (AUC) values, tumor exposure to [3H]TXL was five times greater when administered as PG-TXL than as TXL formulated in Cremophor EL/alcohol vehicle. Furthermore, concentrations of free paclitaxel released from PG-[3H]TXL remained relatively constant in tumor tissue, being 489, 949 and 552 ng/g tumor tissue at 5, 48 and 144 h after dosing, respectively. Autoradiographic images of mice injected with PG-[3H]TXL revealed that radioactivity was primarily located in the periphery of the tumor on day 1 after drug administration and was homogeneously diffused into the center of the tumor by day 6. Over the 144-h study period, [3H]TXL concentrations, predominantly as the inactive conjugate, were higher in tissues with a more abundant reticular endothelial system (i.e. liver, kidney, spleen, lung) than in tissues with less abundant or lacking RE systems (i.e. muscle, fat, brain). Both [3H]TXL and PG-[3H]TXL were excreted primarily through the hepatobiliary route, with a small fraction of each drug (5% and 8.7%, respectively) excreted into the urine within 48 h. CONCLUSIONS: This study indicates that the distribution to tumor tissue was enhanced when [3H]TXL was administered as a macromolecular conjugate, and that free TXL was released and maintained within the tumor for a prolonged period. Thus, the antitumor activity of PG-TXL observed in preclinical studies may be attributed in part to enhanced tumor uptake of PG-TXL.

Phase I study of liposomal annamycin.

Annamycin is a highly lipophilic anthracycline with the ability to bypass the MDR-1 mechanism of cellular drug resistance. In this phase I study, annamycin entrapped in liposomes was administered by a 1- to 2-h intravenous infusion at 3-week intervals. Thirty-six patients with relapsed solid tumors were treated and 109 courses were administered at doses ranging from 3 to 240 mg/m2. The dose-limiting toxicity was thrombocytopenia. Five patients had a probable allergic reaction, requiring discontinuation of treatment in one. Treatment was well tolerated otherwise. No cardiac toxicity was seen on endomyocardial biopsy of four patients studied. There was limited gastrointestinal toxicity and no alopecia. No objective tumor responses were observed. Pharmacokinetic studies at 24, 120 and 240 mg/m2 showed a biexponential plasma concentration-versus-time profile. There was a linear relationship between the dose and the maximal plasma concentration with relatively constant plasma clearance values. The maximum tolerated dose (MTD) for liposomal annamycin defined in this study is 210 mg/m2. Because of a subsequent change in the formulation of the drug, future studies will use 190 mg/m2 as the MTD.

Potentiation of ovarian OCa-1 tumor radioresponse by poly (L-glutamic acid)-paclitaxel conjugate.

PURPOSE: It has been shown that paclitaxel (TXL) can strongly enhance tumor cells' sensitivity to radiation. We examined whether the radiosensitizing effect of paclitaxel can be further enhanced when it is delivered systemically as a polymer-drug conjugate that provides enhanced tumor uptake and prolonged release of TXL in the tumor. METHODS AND MATERIALS: C3Hf/Kam mice bearing 8-mm murine ovarian OCa-1 tumors were treated with i.v.-injected Poly(L-glutamic acid)-paclitaxel (PG-TXL) at an equivalent TXL dose of 80 mg/kg, followed 24 h later by single doses of local radiation ranging from 5 to 15 Gy. To determine how long the radiopotentiation persisted at extended times after PG-TXL administration, mice with OCa-1 tumors were given i.v. PG-TXL and 4, 24, 48, 72, 120, or 168 h later their tumors were irradiated at a dose of 10 Gy. Antitumor activity was determined by delay in tumor growth. Cell cycle distribution was assayed using flow cytometry. Tumor vascular volume was estimated using Tc-99 m-labeled red blood cells. RESULTS: PG-TXL strongly potentiated the radioresponse of the OCa-1 tumor. The enhancement factors ranged from 2.79 to 4.28, depending on radiation dose, when PG-TXL preceded radiation by 24 h. The enhancement factor derived from radiation dose-response curves was as high as 5.13. The radiosensitizing effect of PG-TXL was also dependent on the interval between PG-TXL administration and radiation delivery, with greater enhancement been observed when the interval was decreased. The percentage of G2/M cells was significantly increased to 21.4% 48 h after PG-TXL but declined to a preinjection level of 14.8% 72 h after PG-TXL. PG-TXL only moderately increased the tumor vascular volume by 37% 24 h after PG-TXL administration. CONCLUSION: PG-TXL markedly potentiated response of OCa-1 tumor to radiation. When compared to literature data obtained from the same tumor model used here, PG-TXL exhibited stronger radiosensitization effect than TXL. Although its action is possibly mediated by arrest of cells in G2/M phases of cell cycle and by increased tumor blood supply, PG-TXL may exert its radiopotentiation activity through increased tumor uptake of PG-TXL and sustained release of TXL in the tumor. Our results show that conjugation of TXL to a polymer has the potential to further enhance its radiosensitizing activity and that clinical trials of PG-TXL in combination with radiation is warranted.