Abstract  The activities of base metal oxides comprising 27 % MnO2, 21 % CeO2, and 52 % Al2O3 supported on cordierite monoliths calcined at 1,000 degrees C for 3 h in air dropped very rapidly due to the migration of mobile silicon dioxide (SiO2) from the cordierite to the base metal oxides to react with or physically block the active catalysts. After immobilizing migrating SiO2, cordierite-supported base metal oxides catalysts pretreated with BaO-Al2O3 barrier coats and calcined at 1,000 degrees C for 3 h in air, initiated the oxidation of HCHO, CH3OH, and CO at 150, 170, and 220 degrees C, respectively. These catalysts turned out to be more effective for the formaldehyde oxidation than 0.5 % Pt/Al2O3 precious metal catalysts. The incorporation of small amount of palladium (0.147 wt%) to cordierite-supported base metal oxides pretreated with BaO-Al2O3 barrier coats, showed the superiority for HCHO, CH3OH, and CO oxidation to 0.5 % Pt/Al2O3 precious metal catalysts. The temperatures of 50 % conversion of formaldehyde, methanol, and carbon monoxide were 70 degrees C lower over base metal oxides catalysts than over precious metal catalysts.

Abstract  BIOSIS COPYRIGHT: BIOL ABS. RRM REVIEW ANIMAL ATMOSPHERIC POLLUTANTS HUMORAL IMMUNITY CELLULAR IMMUNITY EXPERIMENTAL MODEL

Abstract  In the initial steps of their metabolic pathway, methanotrophic bacteria oxidize methane to methanol with methane monooxygenases (MMOs) and methanol to formaldehyde with methanol dehydrogenases (MDHs). Several lines of evidence suggest that the membrane-bound or particulate MMO (pMMO) and MDH interact to form a metabolic supercomplex. To further investigate the possible existence of such a supercomplex, native MDH from Methylococcus capsulatus (Bath) has been purified and characterized by size exclusion chromatography with multi-angle light scattering and X-ray crystallography. M. capsulatus (Bath) MDH is primarily a dimer in solution, although an oligomeric species with a molecular mass of ∼450-560 kDa forms at higher protein concentrations. The 2.57 Å resolution crystal structure reveals an overall fold and α2β2 dimeric architecture similar to those of other MDH structures. In addition, biolayer interferometry studies demonstrate specific protein-protein interactions between MDH and M. capsulatus (Bath) pMMO as well as between MDH and the truncated recombinant periplasmic domains of M. capsulatus (Bath) pMMO (spmoB). These interactions exhibit KD values of 833 ± 409 nM and 9.0 ± 7.7 μM, respectively. The biochemical data combined with analysis of the crystal lattice interactions observed in the MDH structure suggest a model in which MDH and pMMO associate not as a discrete, stoichiometric complex but as a larger assembly scaffolded by the intracytoplasmic membranes.

Abstract  Accumulating evidence suggests that cancer-associated stromal fibroblasts (CAFs) contribute to tumor growth by actively communicating with cancer cells. Our aim was to identify the signaling pathways that are involved in tumor-stromal cell interactions in human papillary thyroid carcinoma (PTC). Immunohistochemical analyses were performed with 127 archived formalin-fixed and paraffin-embedded thyroid tissue samples that included 70 cases of PTC, 35 cases of nodular goiter (NG), and 22 cases of normal thyroid tissues. The results showed that the expression levels of Notch1, transforming growth factor beta (TGF-beta 1), and p-Smad3 in PTC cells and alpha-smooth muscle actin (alpha-SMA) in the stroma of PTC were all significantly higher than in NG and normal thyroid tissues. Further analysis showed that in PTC, higher expression levels of Notch1 and TGF-beta 1 were closely related with lymph node metastasis (P < 0.05), whereas for alpha-SMA and p-Smad3, the percent expression increased significantly with advanced tumor stages (P < 0.05). Correlation analysis revealed that TGF-beta 1 expression increased with increased Notch1 and p-Smad3 levels in PTC cells (P < 0.05). Moreover, a significant correlation was found between higher TGF-beta 1 expression in PTC cells and increased alpha-SMA levels in the fibroblasts surrounding the cancer cells (P < 0.05). We identified TGF-beta 1 as an important factor from PTC cells that act in a paracrine manner to influence the activation of stromal fibroblasts. These data suggest that the activation of Notch and TGF-beta/Smad3 pathways in cancer cells influence tumor growth. Moreover, cancer cell-derived-TGF-beta ligands also affect stromal cells in a paracrine fashion and enhance tumor growth.

Abstract  The diverse roles of IGF-1 in physiology include acting as the endocrine intermediate to elicit the anabolic actions of GH. The majority of serum IGF-1 is synthesized in liver, where GH stimulates Igf1 gene transcription via the transcription factor, signal transducer and activator of transcription (Stat)5b. We and others have identified multiple Stat5-binding domains at the Igf1 locus that function in gene regulation, but it remains unclear whether the roles of these domains are tissue specific. Survey of the chromatin landscape of regulatory domains can provide insight about mechanisms of gene regulation, with chromatin accessibility regarded as a hallmark feature of regulatory domains. We prepared chromatin from liver, kidney, and spleen of C57BL/6 mice, and used formaldehyde-associated isolation of regulatory elements to assess chromatin accessibility at the major Igf1 promoter and 7-binding enhancers. Whereas the promoters of other prototypical tissue-specific genes are open in a tissue-specific way, the major Igf1 promoter is open in all 3 tissues, albeit moderately more so in liver. In contrast, chromatin accessibility at Igf1 Stat5-binding domains is essentially restricted to liver, indicating that the enhancers are driving extensive differences in tissue expression. Furthermore, studies with Ghrhr(lit/lit) mice reveal that prior GH exposure is not necessary to establish open chromatin at these domains. Lastly, formaldehyde-associated isolation of regulatory elements of human liver samples confirms open chromatin at IGF1 Promoter 1, but unexpectedly, homologous Stat5-binding motifs are not accessible. We conclude that robust GH-stimulated hepatic Igf1 gene transcription utilizes tissue-specific mechanisms of epigenetic regulation that are established independent of GH signaling.

Abstract  Identifying gene mutations in individual tumors is critical to improve the efficacy of cancer therapy by matching targeted drugs to specific mutations. Gastrointestinal stromal tumors (GIST) are stromal or mesenchymal subepithelial neoplasms affecting the gastrointestinal tract and frequently contain activating gene mutations in either KIT or platelet-derived growth factor A (PDGFRA). Although GIST is highly responsive to several selective tyrosine kinase inhibitors, combined use of inhibitors targeting other mutations is needed to further prolong survival in patients with GIST. In this study, we aim to screen and identify genetic mutations in GIST for targeted therapy using the new Ion Torrent next-generation sequencing platform. Utilizing the Ion Ampliseq Cancer Panel, we sequenced 737 loci from 45 cancer-related genes using DNA extracted from formalin-fixed and paraffin-embedded (FFPE) samples of 121 human gastrointestinal stromal tumors, set up stringent parameters for reliable variant calling by filtering out potential raw base calling errors, and identified frequent mutations in the KIT gene. This study demonstrates the utility of using Ion Torrent sequencing to efficiently identify human cancer mutations. This may provide a molecular basis for clinically developing new drugs targeting these gene mutations for GIST therapy.

Abstract  Context.-Validation of new methodologies for determining human epidermal growth factor receptor 2 gene (HER2/neu) amplification status is crucial for advancing the standard of care and determining treatment for patients with primary and/or metastatic breast carcinoma.

Objective.-To compare results of HER2/neu gene amplification status by 2-color chromogenic in situ hybridization (ISH) on cell block material to HER2/neu status by immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH) in the corresponding resection specimen or previous biopsy specimen.

Design.-Formalin, thrombin, and Cellient cell blocks were prepared from cytologic samples obtained from resection specimens from 27 patients with invasive breast carcinoma. In situ hybridization was performed on cell block sections from 18 of the collected cases, on both the Ventana BenchMark ULTRA and the Ventana BenchMark XT, and the HER2/neu gene amplification status was determined. This was then compared to the HER2/neu status by IHC and/or FISH in the resection specimen or previous biopsy specimen.

Results.-Comparison of HER2/neu status by ISH on the quantifiable cell block sections showed 100% correlation with the HER2/neu status determined by IHC or FISH in the corresponding histologic specimens. The results from thrombin and formalin cell blocks were statistically superior to the results from Cellient cell blocks on both Ventana instruments.

Conclusions.-While further validation and study are needed, preliminary results show that the HER2/neu gene amplification status of breast carcinomas can reliably be determined on thrombin and formalin cell block material by using ISH. More consistent staining and better signal integrity was obtained with the Ventana BenchMark ULTRA than the BenchMark XT.

Abstract  The aim of this study was to analyze protein and gene expression of HER2 in 224 head and neck precancerous and malignant lesions by immunohistochemistry and FISH analysis. In parallel, expression of pStat3, Sox2, IFI16 and p16, Ki67 was evaluated.

Immunohistochemical analysis was assessed on formalin-fixed paraffin-embedded (FFPE) tissue specimens. A combined method for HPV detection consisting of p16 immunostaining and two PCR probes was applied. HER2 gene status was evaluated by FISH analysis.

HPV DNA was detected in 24% of cases with predominant HPV16 genotype. HPV-positive lesions had higher HER2, pStat3 and within carcinoma group, and higher IFI16 expression compared to the HPV-negative group (Fig. 1A-B-C). A strong positive correlation between Sox2 and proliferative activity was observed, whereas IFT16 expression displayed a negative relationship with Sox2 and Ki67 activity. The most striking result was higher pStat3 expression in HPV-positive lesions and its strong positive correlation with IFT16 expression. The presence of HPV may induce upregulation of HER2/neu, pStat3 and IFI16. High levels and a strong positive correlation between pStat3 and IFI16 suggest their synergistic pro-apoptotic effects in HPV-positive lesions.

Abstract  Background: Gene expression signatures have proven to be useful tools in many cancers to identify distinct subtypes of disease based on molecular features that drive pathogenesis, and to aid in predicting clinical outcomes. However, there are no current signatures for kidney cancer that are applicable in a clinical setting.

Objective: To generate a signature biomarker for the clear cell renal cell carcinoma (ccRCC) good risk (ccA) and poor risk (ccB) subtype classification that could be readily applied to clinical samples to develop an integrated model for biologically defined risk stratification.

Design, setting, and participants: A set of 72 ccRCC sample standards was used to develop a 34-gene classifier (ClearCode34) for assigning ccRCC tumors to subtypes. The classifier was applied to RNA-sequencing data from 380 nonmetastatic ccRCC samples from the Cancer Genome Atlas (TCGA), and to 157 formalin-fixed clinical samples collected at the University of North Carolina.

Outcome measurements and statistical analysis: Kaplan-Meier analyses were performed on the individual cohorts to calculate recurrence-free survival (RFS), cancer-specific survival (CSS), and overall survival (OS). Training and test sets were randomly selected from the combined cohorts to assemble a risk prediction model for disease recurrence.

Results and limitations: The subtypes were significantly associated with RFS (p < 0.01), CSS (p < 0.01), and OS (p < 0.01). Hazard ratios for subtype classification were similar to those of stage and grade in association with recurrence risk, and remained significant in multivariate analyses. An integrated molecular/clinical model for RFS to assign patients to risk groups was able to accurately predict CSS above established, clinical risk-prediction algorithms.

Conclusions: The ClearCode34-based model provides prognostic stratification that improves upon established algorithms to assess risk for recurrence and death for nonmetastatic ccRCC patients.

Patient summary: We developed a 34-gene subtype predictor to classify clear cell renal cell carcinoma tumors according to ccA or ccB subtypes and built a subtype-inclusive model to analyze patient survival outcomes. (C) 2014 European Association of Urology. Published by Elsevier B.V. All rights reserved.

Abstract  The aim of this paper was to analyze the role of melatonin (MLT) in acute and chronic pain states, and to describe the profile of melatonin as a potential analgesic. Melatonin (N-acetyl-5-methoxytryptamine) is the main pineal neurohormone involved in many physiological and endocrine functions, as well as in behavioral states in humans and animals. One may mention functions such as the regulation of circadian rhythm, sleep, mood, processes of reproduction, immune response, and aging. The distribution of MLT receptors in the spinal cord and in many other regions of the brain associated with receiving pain impulses, indirectly confirms the possible involvement of indoleamine in the phenomenon of pain. In clinical studies, MLT has been shown to have analgesic benefits in patients with acute and chronic pain (migraine, fibromyalgia, irritable bowel syndrome). In preliminary studies, it has also been suggested that MLT may have potent analgesic and anxiolytic properties in the perioperative period.

Abstract  Formaldehyde (FA) ranks 25th in the overall U.S. chemical production, with more than 5 million tons produced each year. Given its economic importance and widespread use, many people are exposed to FA occupationally. Recently, based on the correlation with nasopharyngeal cancer in humans, the International Agency for Research on Cancer (IARC) confirmed the classification of FA as a Group I substance. Considering the epidemiological evidence of a potential association with leukemia, the IARC has concluded that FA can cause this lymphoproliferative disorder. Our group has developed a method to assess the exposure and genotoxicity effects of FA in two different occupational settings, namely FA-based resins production and pathology and anatomy laboratories. For exposure assessment we applied simultaneously two different techniques of air monitoring: NIOSH Method 2541 and Photo Ionization Detection Equipment with simultaneously video recording. Genotoxicity effects were measured by cytokinesis-blocked micronucleus assay in peripheral blood lymphocytes and by micronucleus test in exfoliated oral cavity epithelial cells, both considered target cells. The two exposure assessment techniques show that in the two occupational settings peak exposures are still occurring. There was a statistical significant increase in the micronucleus mean of epithelial cells and peripheral lymphocytes of exposed individuals compared with controls. In conclusion, the exposure and genotoxicity effects assessment methodologies developed by us allowed to determine that these two occupational settings promote exposure to high peak FA concentrations and an increase in the micronucleus mean of exposed workers. Moreover, the developed techniques showed promising results and could be used to confirm and extend the results obtained by the analytical techniques currently available.

Abstract  A new simple and highly sensitive catalytic kinetic method for the determination of trace amount of FA in food sample has been established. The method was based on the catalytic effect of FA on the oxidation of eosin Y by potassium bromate in present of phosphoric acid. The reaction was monitored spectrophotometrically by measuring the decrease in absorbance of eosin Y at 518 nm. Under the optimized experimental conditions, the developed method allowed the determination of FA in the range of 0.03-0.6 mu g mL(-1) with a good precision, and the limit of detection was down to 0.009881 mu g mL(-1). The relative standard deviation of five replicate measurements for the determination of FA in concentration 0.12 mu g mL(-1) was 1.8%. The proposed method was successfully applied to the determination of FA in food directly and satisfactory results were obtained. (C) 2014 Elsevier B.V. All rights reserved.

Abstract  Adsorption of formaldehyde (H2CO) on B-, N-, Si-, Al-, Cr-, Mn-, and Au-doped graphene was theoretically studied using first-principles approach based on density functional theory in order to exploit their potential applications as H2CO gas sensors. The electronic and magnetic properties of the graphene-molecule adsorption adducts are strongly dependent on the dopants. H2CO molecule is adsorbed weakly on Band N-doped graphene; in general, strong chemisorption is observed on Si-, Al-, Cr-, Mn-, and Au-doped graphene. The most stable adsorption geometries, adsorption energies, magnetic moments, charge transfers, and density of states of these systems are thoroughly discussed. This work reveals that the sensitivity of graphene-based chemical gas sensors for H2CO can be drastically improved by introducing appropriate dopant. Al and Mn are found to be the best choices among all the dopants. (C) 2014 Elsevier B.V. All rights reserved.

Abstract  In this work, we present a novel process for the fabrication of a silver-nanoparticle-modified electrode using silver ion implantation. This method is facile, low-cost and environmental friendly without the use of any linking chemicals. The modified electrode was verified by scanning electron microscope (SEM), atomic force microscope (AFM), X-ray photoelectron spectroscopy (XPS), electrochemical impedance spectra (EIS) and cyclic voltammetry (CV). The AgNPs formed on the electrode are in the zero-valent metallic state with a size distribution in the range of 3-6 nm. The modified electrode shows prominent electrocatalytic activity towards the oxidation of formaldehyde with long-term stability and could be useful in fuel cells. (C) 2014 Elsevier B.V. All rights reserved.

Abstract  Catalytic oxidation of formaldehyde (HCHO) to CO2 at ambient conditions is of great interest for indoor HCHO purification. Here, we report that sodium-doped Pd/TiO2 is a highly effective catalyst for the catalytic oxidation of HCHO at room temperature. It was observed that Na doping has a dramatic promotion effect on the Pd/TiO2 catalyst and that nearly 100% HCHO conversion could be achieved over the 2Na-Pd/TiO2 catalyst at a GHSV of 95000 h(-1) and HCHO inlet concentration of 140 ppm at 25 degrees C. The mechanism of the Na-promotion effect was investigated by using Brunauer-Emmett-Teller (BET), X-ray diffraction (XRD), CO chemisorption, Temperature-programmed reduction by H-2 (H-2-TPR), X-ray photoelectron spectroscopy (XPS) and temperature-programmed desorption of O-2 (O-2-TPD) methods. The results showed that Na species addition can induce and further stabilize a negatively charged and well-dispersed Pd species, which then facilitates the activation of H2O and chemisorbed oxygen, therefore resulting in the high performance of the 2Na-Pd/TiO2 catalyst for the ambient HCHO destruction.

Abstract  An effect of an ammonia solution concentration, temperature, l: s ratio on a MoO3 extraction process was revealed in studying ammonia leaching of molybdenum oxide from spent iron-molybdenum catalyst of oxidation of methanol to formaldehyde The data obtained allow to optimize the extraction process of MoO3 from the spent Fe-Mo catalysts.

Abstract  Zinc oxide (ZnO) and Fe doped ZnO nanorods were fabricated on ceramic balls by a simple low cost wet-chemical route. The as-prepared samples were characterized by XRD, SEM, TEM and UV-Vis spectroscopy. Photocatalytic activities of pure ZnO and Fe doped ZnO nanorods were investigated for degradation of formaldehyde gas under UV irradiation. The results show that doped catalyst exhibits superior photocatalytic performance than that of pure ZnO nanorods and 4% Fe doping catalyst displays the optimal photoactivity. The catalyst has good stability and is easy to be recoverd. The catalytic efficiency still maintains 85% after 10 cycls.

Abstract  A solid-liquid extraction method is developed to establish the contents of selenium in breast cancer biopsies. The method is based on the ultrasound-assisted extraction of selenium from pretreated biopsies prior to Se determination by atomic absorption spectrometry with longitudinal-Zeeman background correction. Fifty-one breast biopsies were collected from the Cies Hospital (Vigo, Spain), 32 of which correspond to tumor tissue and 19 to normal tissue (parenchyma). Difficulties arising from the samples analyzed, i.e. small samples mass (50-100 mg), extremely low Se contents and sample texture modification including tissue hardening due to formaldehyde preservation are addressed and overcome. High intensity sonication using a probe together with addition of hydrogen peroxide succeeded in completely extracting Se from biopsies. The multiple injection technique was useful to tackle the low Se contents present in some biopsies. The detection limit was 25 ng g(-1) of Se and the precision, expressed as relative standard deviation, was less than 10%. Se contents ranged from 0.08 to 0.4 mu g g(-1) for parenchyma samples and from 0.09 to 0.8 mu g g(-1) for tumor samples. In general, Se levels in tumor biopsies were higher as compared with the adjacent normal tissue in 19 patients by a factor of up to 6. Analytical data confirmed Se accumulation in the breast tumors. (c) 2006 Elsevier B.V. All rights reserved.

Abstract  The electrochemical redox behavior of the polynuclear mixed valence ruthenium oxide cyanometallate complexes (mvRuOx-MCN, M=Fe, Cr, Ni, Cu, Ru and Pt) have been systematically studied in this report by using three redox sensitive organic probes of glucose, ethanol and formaldehyde. The results were interpreted by the well-established ruthenium oxide and Prussian blue chemistry. The mvRuOx-MCN, under the category of Ru-based Prussian blue analogue, was found to possess superior electrocatalytic activity than either ruthenium oxide or Prussian blue in acidic mediums. The electrogenerated oxy/hydroxy-Ru-VII state (at + 1.1 V vs. Ag/AgCl) was unusually stabilized in the mvRuOx-MCN matrix without any disproportion reaction in acidic environments. In contrast to those of earlier studies, possible structure in terms of the -Ru-III/II-NC-M-and -Ru-III/II-O-Ru-VII/VI-sites was proposed here. Enzyme-less analytical detection of glucose in acidic conditions was first time demonstrated with sensitivity comparable to that of ruthenium oxide-based electrodes in alkaline solutions.

Abstract  Levels of formaldehyde, formed within the cells of plants, animals, and man, are related to the physiological state of an organism. We have previously reported the possibility of determining the formaldehyde content of human teeth in some physiological states, and it was established that the level of HCHO in carietic teeth is higher than in healthy teeth. The aim of the work described in this paper was the OPLC determination of HCHO levels in teeth suffering from eight different rare pathological disorders, It was shown that significantly different levels of formaldehyde were present in teeth in different pathological states. The highest HCHO content was observed in teeth with acute inflammatory changes. The Lowest Level was found in teeth with no inflammatory changes.

Abstract  Purpose: To determine whether the ultrastructural changes in the eyes of an animal model of Vogt Koyanagi-Harada (VKH) disease are similar to those in human eyes with VKH disease.

Methods: Pigmented rats derived by crossing brown Norway and Lewis rats were immunized with tyrosinase-family proteins, and the eyes were enucleated after different postinoculation times. The eyes were fixed in glutaraldehyde or formaldehyde and prepared for light or electron microscopy (EM).

Results: EM showed that thickening of the choroid resulted from the invasion of many lymphocytes and epithelioid-like cells. The melanocytes were surrounded by lymphocytes and lost their normal spindle shape. Neutrophils and eosinophils were also present in the choroid. These findings are very similar to the changes reported in eyes from patients with VKH disease or sympathetic ophthalmia.

Conclusions: The similarity of the ultrastructural changes in this model to those in human VKH disease indicate that this animal model can serve as a histopathological model of human VKH disease. (C) Japanese Ophthalmological Society 2004.

Abstract  Phenol formaldehyde resin (PFR) was coated on the surface of carbon nanotubes (CNTs) by a simple one-pot hydrothermal approach. The coating did not ruin the nanotube sidewall, and its shell thickness was finely controlled on the nanometer scale by adjusting the amount of monomer. Silver nanoparticles were attached onto the PFR@CNT composites followed by the in-situ redox of AgNO(3) and as-prepared PFR/CNTs through hydrothermal treatment. (C) 2011 Elsevier Masson SAS. All rights reserved.

Abstract  The occurrence of tetrahydro-beta-carboline-3-carboxylic acids (TH beta C-3-COOHs) in foodstuffs was investigated. Spectral and chromatographic data showed the occurrence of 1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (THCA) and 1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (MTCA) in foodstuffs. The content of TH beta C-3-COOHs given as a sum of both THCA and MTCA was as follows: soy sauce, 94-517 mg/L; seasoning, 0.75-32.8 mu g/g; table wine, 1.7-6.6 mg/L; beer, 0.3-17.9 mg/L; cider, 0.06-0.2 mg/L; liquor, 0-7.6 mg/L; nine vinegar, 3.9-9.7 mg/L; cider vinegar, 0.19-1 mg/L; yogurt, 0.05-0.15 mu g/g; cheese, 0-3.4 mu g/g; soft drinks, 0-0.45 mg/L; fruit juices, 0.1-5.1 mg/L; smoked fish, 0.08-0.4 mu g/g; and bread, 0.16-3 mu g/g. Usually MTCA was the major substance within TH beta C-3-COOHs, but bread and smoked fish contained more THCA. Experiments in which foodstuffs were spiked with formaldehyde and acetaldehyde proved the chemical formation of THCA and MTCA, respectively. It is concluded that the exogenous intake of these substances during the human ingestion of foods may be partially responsible of the reported endogenous presence of TH beta C-3-COOHs in the human biological tissues and fluids.

Abstract  Lipid peroxidation (LPO) is the oxidative deterioration of polyunsaturated fatty acids (PUFA) with the production of lipid hydroperoxides, cyclic peroxides, cyclic endoperoxides, and finally fragmentation to ketones and aldehydes (including malonaldehyde, MDA). Estimation of LPO through MDA formation measured by assaying thiobarbituric acid (TBA) reactive products remains the method of choice to study the development of oxidative stress in tissues. However, MDA estimation by TBA reactive products is non-specific and often gives erroneous results. In this report we describe a method using high-performance liquid chromatographic separation to estimate MDA, formaldehyde (FDA), acetaldehyde (ADA), acetone, and propionaldehyde (PDA), the degradation products of oxygen-derived free radicals (ODFR) and PUFA, as presumptive markers for LPO. Oxidative stress was induced in the tissue by perfusing an isolated rat heart with hydroxyl radical generating system (xanthine + xanthine oxidase + FeCl3 + EDTA). The coronary effluents were collected, derivatized with 2,4-dinitrophenylhydrazine (DNPH), and extracted with pentane. Aliquots of 25 mu l in acetonitrile were injected onto a Beekman Ultrasphere C-18 (3 mu m) column. The products were eluted isocratically with a mobile phase containing acetonitrile-water-acetic acid (40:60:0.1, v/v/v), measured at three different wavelengths (307, 325 and 356 nm) using a Waters M-490 multichannel UV detector and collected for gas chromatography-mass spectrometry (CC-MS) analysis. The peaks were identified by cochromatography with DNPH derivatives of authentic standards, peak addition, UV pattern of absorption at the three wavelengths, and by CC-MS. The retention items of MDA, FDA, ADA, acetone, and PDA were 5.3, 6.6, 10.3, 16.5, and 20.5 min, respectively. The results of our study indicated progressive increase of all five lipid metabolites as a function of the duration of ODFR perfusion. Hydroxyl radical scavengers, superoxide dismutase plus catalase, completely inhibited the formation of these lipid metabolites, demonstrating that the release of lipid metabolites from the isolated heart was indeed in response to oxidative stress. Since MDA, FDA, ADA, acetone, and PDA are the products of ODFR-PUFA interactions, this method allows proper estimation of LPO which monitors the oxidative stress developed during the reperfusion of ischemic myocardium.

Abstract  Purpose: CD52 is a GPI-linked glycoprotein expressed by B cells, T cells, monocytes, and macrophages. The humanized monoclonal antibody alemtuzumab (CAMPATH-1H) is specific for CD52 and is Food and Drug Administration - approved for the treatment of relapsed or refractory chronic lymphocytic leukemia (CLL). The utility of CAMPATH in the treatment of other hematologic neoplasms has been explored; however, a comprehensive survey of CD52 expression among a broad spectrum of WHO-defined tumor types has not been completed.

Experimental Design: We evaluated 294 hematologic neoplasms for the presence of CD52 using standard immunohistochemical techniques on paraffin-embedded biopsy specimens fixed with formalin, B-Plus, Zenker's acetic acid, or B5-formalin.

Results: The vast majority of low-grade B cell lymphoproliferative disorders (CLL/small lymphocytic leukemia, follicular lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, and mucosa-associated lymphoid tissue lymphomas) express CD52. In addition, we found that the majority of precursor B cell acute lymphoblastic leukemia/lymphomas express this antigen. In contrast, there is surprising heterogeneity in CD52 expression among more aggressive B cell lymphomas, with 25% of cases of diffuse large B cell lymphoma and Burkitt lymphoma demonstrating no detectable CD52. In addition, the majority of neoplasms of the T cell lineage are negative for the antigen, including most cases of precursor T cell acute lymphoblastic leukemia/ lymphoma, anaplastic large cell lymphoma, and peripheral T cell lymphoma, not otherwise specified. Finally, the vast majority of cases of acute myeloid leukemia, Hodgkin lymphoma, and multiple myeloma are negative for CD52 expression.

Conclusion: In contrast with CLL, the variable expression of CD52 among other hematologic malignancies suggests that target validation on a case-by-case basis will likely be necessary to guide the rational analysis of CAMPATH therapy.