The expression of HSP60 and HSP10 in large bowel carcinomas with lymph node metastase

Background

Cancer serum protein profiling by mass spectrometry has uncovered mass profiles that are potentially diagnostic for several common types of cancer. However, direct mass spectrometric profiling has a limited dynamic range and difficulties in providing the identification of the distinctive proteins. We hypothesized that distinctive profiles may result from the differential expression of relatively abundant serum proteins associated with the host response.

Methods

Eighty-four antibodies, targeting a wide range of serum proteins, were spotted onto nitrocellulose-coated microscope slides. The abundances of the corresponding proteins were measured in 80 serum samples, from 24 newly diagnosed subjects with lung cancer, 24 healthy controls, and 32 subjects with chronic obstructive pulmonary disease (COPD). Two-color rolling-circle amplification was used to measure protein abundance.

Results

Seven of the 84 antibodies gave a significant difference (p < 0.01) for the lung cancer patients as compared to healthy controls, as well as compared to COPD patients. Proteins that exhibited higher abundances in the lung cancer samples relative to the control samples included C-reactive protein (CRP; a 13.3 fold increase), serum amyloid A (SAA; a 2.0 fold increase), mucin 1 and α-1-antitrypsin (1.4 fold increases). The increased expression levels of CRP and SAA were validated by Western blot analysis. Leave-one-out cross-validation was used to construct Diagonal Linear Discriminant Analysis (DLDA) classifiers. At a cutoff where all 56 of the non-tumor samples were correctly classified, 15/24 lung tumor patient sera were correctly classified.

Conclusion

Our results suggest that a distinctive serum protein profile involving abundant proteins may be observed in lung cancer patients relative to healthy subjects or patients with chronic disease and may have utility as part of strategies for detecting lung cancer.     

Effect of muscarinic receptor blockade on canine gastric tone and compliance in vivo.

Muscarinic pathways are involved in maintaining gastric tone during fasting and atropine is known to decrease gastric tone via blockade of a tonic vagal cholinergic input. Our aim was to assess the role of different muscarinic receptors in modulating canine gastric tone and compliance in vivo by using "selective" muscarinic receptor antagonists (telenzepine, AF-DX 116 and 4-DAMP for M1, M2, and M3 receptors, respectively) and the non-selective muscarinic receptor antagonist atropine. In four fasting, conscious dogs, we characterized the pressure-volume relationship in the proximal stomach by using a barostat. Drug effects were investigated by studying pressure-volume relationships before and 15 min after intravenous administration telenzepine, AF-DX 116, 4-DAMP or atropine. Pressure-volume curves were fitted by non-linear regression analysis. Before drug administration, the curve that best fitted the pressure-volume relationship was exponential. Atropine (100 microg kg-1) immediately decreased baseline gastric tone, i.e. relaxed the stomach (Deltavolume at 2 mmHg=236+/-15 ml; P<0.05), and significantly (P<0.01) shifted the pressure-volume curve to the left. Telenzepine, at the lowest dose (3 microg kg-1), shifted the pressure-volume curve to the right (P<0.01). AF-DX 116 at the lower dose (422 microg kg-1) had no effect on baseline gastric tone or the gastric pressure-volume curve, whereas the higher dose (2532 microg kg-1) significantly shifted the pressure-volume curve to the left (P<0.01), but did not increase baseline gastric volume. Finally, 4-DAMP (13.5, 45, 135 microg kg-1) immediately decreased baseline gastric tone (Deltavolume at 2 mmHg=97+/-29 ml, 110+/-35 ml and 155+/-21 ml, respectively) and significantly shifted the pressure-volume curve to the left (P<0.01). We conclude that muscarinic pathways are involved in modulating canine gastric tone and compliance during fasting: M3 receptors seem to play a key role in excitatory pathways, whereas the shift of pressure-volume curve to the right observed with the lowest dose of telenzepine is consistent with the existence of M1 receptors on inhibitory pathways.     

Macrophage depletion induced by clodronate-loaded erythrocytes

Given the important role of macrophages in various disorders, the transient and organ specific suppression of their functions may benefit some patients. Until now, liposome-encapsulated bisphosphonate clodronate has been extensively proposed to this end. In this paper, we demonstrate that erythrocytes loaded with clodronate can also be effective in macrophage depletion. Here, clodronate was encapsulated in erythrocytes through hypotonic dialysis, isotonic resealing and reannealing to final concentrations of 4.1 +/- 0.4 and 10.1 +/- 0.8 micromol/ml of human and murine erythrocytes, respectively. The ability of clodronate-loaded erythrocytes to deplete macrophages was evaluated both in vitro and in vivo. In vitro studies on human macrophages showed that a single administration of engineered erythrocytes was able to reduce cell adherence capacity in a time-dependent manner, reaching 50 +/- 4% reduction, 13 days post treatment. The administration of loaded erythrocytes to cultures of murine peritoneal macrophages was able to reduce macrophage adhesion 67 +/- 3%, 48 h post treatment. In vivo, the ability of clodronate-loaded erythrocytes to deplete macrophages was evaluated both in Swiss and C57BL/6 mice. Swiss mice received 125 microg of clodronate through erythrocytes and 6 days post treatment 69 +/- 7% reduction in the number of adherent peritoneal macrophages and 75 +/- 5% reduction in number of spleen macrophages were observed. C57BL/6 mice received 220 microg clodronate by RBC and 3 and 8 days post treatment 65 +/- 7% reduction in the number of spleen macrophages and the complete depletion of liver macrophages were obtained. In summary, our results indicate that clodronate selectively targeted to the phagocytic cells by a single administration of engineered erythrocytes is able to deplete macrophages, even if not completely. The transient suppression of macrophage functions through clodronate-loaded erythrocytes can be used in many biomedical phenomena and research applications.     

HPMA copolymer-aminoglutethimide conjugates inhibit aromatase in MCF-7 cell lines

N-(2-Hydroxypropyl)methacrylamide (HPMA) copolymer-doxorubicin (Dox) has already shown clinical activity in breast cancer patients. Moreover, we have recently found that an HPMA conjugate containing a combination of both Dox and the aromatase inhibitor aminoglutethimide (AGM) shows significantly increased anti-tumour activity in vitro. To better understand the mechanism of action of HPMA copolymer-AGM conjugates several models were used here to investigate their effect on cell growth and aromatase inhibition. Cytotoxicity of HPMA copolymer conjugates containing AGM, Dox and also the combination AGM-Dox was determined by MTT assay in MCF-7 and MCF-7ca cells. Androstenedione (5 x 10(- 8) M) stimulates the growth of MCF-7ca cells. Both free AGM and polymer-bound AGM (0.2-0.4 mg/ml) were shown to block this mitogenic activity. When MCF-7ca cells were incubated [(3)H]androstenedione both AGM and HPMA copolymer-GFLG-AGM (0.2 mg/ml AGM-equiv.) showed the ability to inhibit aromatase. Although, free AGM was able to inhibit isolated human placental microsomal aromatase in a concentration dependent manner, polymer-bound AGM was not, suggesting that drug release is essential for activity of the conjugate. HPMA copolymer conjugates containing aromatase inhibitors have potential for the treatment of hormone-dependant cancers, and it would be particularly interesting to explore further as potential therapies in post-menopausal women as components of combination therapy.     

The role of newer antiglutamatergic agents as mood-stabilizers in bipolar disorder

In bipolar patients, maintenance treatment with anticonvulsive agents is a valid alternative to lithium. These agents have widely varying mechanisms of action. Some of these medications focus on the current understanding of antiglutamatergic mechanisms of action and their treatment implications for bipolar disorders.     

Antiangiogenesis by chemotherapeutic agents

Many tumors are not curable because current treatments primarily target the tumor cells. Intratumoral endothelial cells, on the other hand, proliferate rapidly and are sensitive to the cytotoxic effects of chemotherapeutic agents. This review summarizes the literature concerning the antiangiogenic effects of these agents when administered alone or in combination with other angiogenesis inhibitors.     

Revised analyses of the National Morbidity, Mortality, and Air Pollution Study: mortality among residents of 90 cities

This article presents findings from updated analyses of data from 90 U.S. cities assembled for the National Morbidity, Mortality, and Air Pollution Study (NMMAPS). The data were analyzed with a generalized additive model (GAM) using the gamfunction in S-Plus (with default convergence criteria previously used and with more stringent criteria) and with a generalized linear model (GLM) with natural cubic splines. With the original method, the estimated effect of PM(10) (particulate matter 10 microm in mass median aerodynamic diameter) on total mortality from nonexternal causes was a 0.41% increase per 10-microg/m(3) increase in PM(10); with the more stringent criteria, the estimate was 0.27%; and with GLM, the effect was 0.21%. The effect of PM(10) on respiratory and cardiovascular mortality combined was greater, but the pattern across models was similar. The findings of the updated analysis with regard to spatial heterogeneity across the 90 cities were unchanged from the original analyses.     

Multisite time-series studies versus cohort studies: methods, findings, and policy implications

Time-series and cohort studies of air pollution on human health have advanced greatly our understanding of the effects of air pollution on health since the earliest studies. Availability of large national databases and progress in computational tools and statistical methods have made possible the estimation of national average pollution effects and the exploration of potential sources of heterogeneity in the effects of air pollution across countries or regions. Interpretation of the findings needs to account for several challenges, including confounding and the resolution of seemingly conflicting results from time-series and cohort studies. This article presents an overview of the time-series and cohort studies' approaches for estimating the relative risk of mortality from particulate air pollution and discusses the statistical issues and challenges inherent in each of these studies. We also discuss policy relevant summaries in air pollution epidemiology, approaches for estimating the impact of particulate matter on mortality from time-series and cohort studies, and research opportunities under the National Medicare Cohort Study (NMCS).     

Testing the harvesting hypothesis by time-domain regression analysis. I: baseline analysis

Although the association between air pollution and daily mortality is well established, the mechanisms by which air pollution results in excess mortality are not yet well understood. In particular, there exists debate over whether air pollution has a direct effect on mortality in the general population or simply shortens the life span of frail individuals, a hypothesis referred to as "harvesting." The goal of this investigation is to test the harvesting hypothesis using the time-domain regression method of Dominici et al. (2003a). We conducted simulations based on a two-compartment model that divides the population into a larger group of healthy individuals and a frail subpopulation. Death from air pollution is assumed to take place in two steps, by first moving from healthy population to the frail pool, then death with probability related to the level of air pollution. Using time-domain analysis, we seek to identify data patterns that would be characteristic of harvesting under different scenarios. For a pure harvesting model, time-domain analysis indicates that mortality is associated with a short-term air pollution episode of less than 2 d if the mean residency time in the frail pool is short. If both entrants and deaths depend on the level of air pollution and the rates of entry to and exit from the frail pool are about the same, the log relative risk estimates are essentially unchanged at all time scales. If pollution affects mortality in the frail pool more than entrants, larger effects will occur at shorter time scales.     

Diclofenac salts. III. Alkaline and earth alkaline salts

Diclofenac salts containing the alkaline and two earth alkaline cations have been prepared and characterized by scanning electron microscopy (SEM) and EDAX spectroscopy; and by thermal and thermogravimetric analysis (TGA): all of them crystallize as hydrate when precipitated from water. The salts dehydrate at room temperature and more easily on heating, but recovery the hydration, when placed in a humid environment. X-ray diffraction spectra suggest that on dehydration new peaks appear on diffractograms and the lattice of the salts partially looses crystallinity. This phenomenon is readily visible in the case of the calcium and magnesium salts, whose thermograms display a crystallization exotherm, before melting or decomposing at temperatures near or above 200 degrees C; these last salts appear to form solvates, when prepared from methanol. The thermogram of each salt shows a complex endotherm of dehydration about 100 degrees C; the calcium salt displays two endotherms, well separated at about 120 and 160 degrees C, which disappear after prolonged heating. Decomposition exotherms, before or soon after the melting, appear below 300 degrees C. The ammonium salt is thermally unstable and, when heated to start dehydration, dissociates and leaves acidic diclofenac.