Bax expression is predictive of favorable clinical outcome in chemonaive advanced gastric cancer patients treated with capecitabine, oxaliplatin, and irinotecan regimen

OBJECTIVE: Bax protein is a key mediator of apoptosis, and it might be related to chemosensitivity. The purpose of this study was to evaluate the prognostic role of Bax in patients with advanced gastric cancer treated with triplet chemotherapy COI regimen (capecitabine, oxaliplatin, and irinotecan). METHODS: Pretreatment tissue blocks were available for 23 consecutive patients, selected for good performance status (ECOG ≤ 1) and consenting for treatment with first-line COI at a single institution. Bax levels were classified as positive or negative by immunohistochemistry (bax N20; Santa Cruz Biotechnology) and related to outcome in terms of response rate, progression-free survival, and overall survival. RESULTS: Bax-negative and -positive samples were 26% and 74%, respectively. Bax expression was associated with significantly higher response rate (87% vs 33%), progression-free survival (8.7 vs 4.9 months, P = .016), and overall survival (23.8 vs 12.7 months, P = .025). In multivariate analysis including Bax and performance status, low Bax independently predicted worse outcome, along with suboptimal performance status. CONCLUSIONS: In advanced gastric cancer, Bax expression was related to clinical benefit with COI regimen. Whether Bax is a prognostic or mixed prognostic/predictive factor warrants prospective confirmation. It is to be defined if Bax predicts sensitivity to platinum analogs or to whatever chemotherapy regimen.     

Marie Ménard apples with high polyphenol content and a low-fat diet reduce 1,2-dimethylhydrazine-induced colon carcinogenesis in rats: Effects on inflammation and apoptosis

Inflammation may increase cancer risk, therefore, we studied whether polyphenol-rich Marie Ménard (MM) apples with reported anti-inflammatory activity prevent 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis in rats and, likewise whether high-fat (HF) diet promoting carcinogenesis, may affect inflammation. DMH-induced rats were fed for 15 weeks with: an HF diet (23% corn oil w/w); an HF diet containing 7.6% w/w lyophilized MM (apple diet (AD)); a low-fat (LF) diet and an HF diet containing piroxicam (PXC) (0.01% w/w) as control. Mucin depleted foci (MDF), precancerous lesions in the colon, were dramatically reduced in the AD, LF, and PXC groups compared with the HF. Peritoneal macrophage activation, an index of systemic inflammation, was significantly decreased in the AD, LF, and PXC groups. TNF-α, iNOS, IL-1β, IL-6 m-RNA expression in the colon, as well as CD68 cells and plasmatic PGE2 were lower in the AD, but not in the LF group. Apoptosis in the MDF of both the AD and LF-fed rats was significantly higher than in HF rats. In conclusion, AD has a strong chemopreventive effect, reducing inflammation, and increasing apoptosis, while the chemopreventive effect of the LF diet seems mediated mainly by increased apoptosis in MDF.     

Diastolic time – frequency relation in the stress echo lab: filling timing and flow at different heart rates

 

A cutaneous force-frequency relation recording system based on first heart sound amplitude vibrations has been recently validated. Second heart sound can be simultaneously recorded in order to quantify both systole and diastole duration.

Aims

1- To assess the feasibility and extra-value of operator-independent, force sensor-based, diastolic time recording during stress.

Methods

We enrolled 161 patients referred for stress echocardiography (exercise 115, dipyridamole 40, pacing 6 patients). The sensor was fastened in the precordial region by a standard ECG electrode. The acceleration signal was converted into digital and recorded together with ECG signal. Both systolic and diastolic times were acquired continuously during stress and were displayed by plotting times vs. heart rate. Diastolic filling rate was calculated as echo-measured mitral filling volume/sensor-monitored diastolic time.

Results

Diastolic time decreased during stress more markedly than systolic time. At peak stress 62 of the 161 pts showed reversal of the systolic/diastolic ratio with the duration of systole longer than diastole. In the exercise group, at 100 bpm HR, systolic/diastolic time ratio was lower in the 17 controls (0.74 ± 0.12) than in patients (0.86 ± 0.10, p < 0.05 vs. controls). Diastolic filling rate increased from 101 ± 36 (rest) to 219 ± 92 ml/m²* s^-1 at peak stress (p < 0.5 vs. rest).

Conclusion

Cardiological systolic and diastolic duration can be monitored during stress by using an acceleration force sensor. Simultaneous calculation of stroke volume allows monitoring diastolic filling rate. Stress-induced "systolic-diastolic mismatch" can be easily quantified and is associated to several cardiac diseases, possibly expanding the spectrum of information obtainable during stress.     

Oxygen uptake during mineralization of photosynthesized carbon from phytoplankton of the Barra Bonita reservoir: a mesocosm study.

This study aimed to discuss and describe the oxygen consumption during aerobic mineralization of organic products (cells and excretion products) from five unialgal cultures: Cryptomonas sp., Microcystis aeruginosa, Anabaena spiroides, Thalassiosira sp. and Aulacoseira granulata. These species were isolated from Barra Bonita reservoir (22 degrees 29' S and 48 degrees 34' W) and cultivated in the laboratory. From each culture, two decomposition chambers were prepared; each chamber contained about 130 mg.L(-1) of carbon from water samples of the reservoir. The chambers were aerated and incubated in the dark at 20.0 degrees C. The concentration of dissolved oxygen, pH values and electrical conductivity of the solutions were determined during a period of 10 days. The results indicated increases in oxygen consumption for all the solutions studied and also for electrical conductivity. The pH values presented a decreasing tendency throughout the experiment. Oxygen consumption varied from 43 (Aulacoseira granulata chamber) to 345 mg O2 g(-1) C (Anabaena spiroides chamber). Decrease in pH values was probably due to increase in CO2 concentration from microbial respiration. Increase in electrical conductivity might be due to the liberation of ions during decomposition. The results demonstrate the potentiality of the studied genera in influencing oxygen availability followed by a die-off event. It also indicates the possibility of changing of the electrical conductivity and pH values in the water column due the aerobic algae mineralization.     

beta-amyloid activates the O-2 forming NADPH oxidase in microglia, monocytes, and neutrophils. A possible inflammatory mechanism of neuronal damage in Alzheimer's disease.

The deposition of beta-amyloid in the brain is the key pathogenetic event in Alzheimer's disease. Among the various mechanisms proposed to explain the neurotoxicity of beta-amyloid deposits, a new one, recently identified in our and other laboratories, suggests that beta-amyloid is indirectly neurotoxic by activating microglia to produce toxic inflammatory mediators such as cytokines, nitric oxide, and oxygen free radicals. Three findings presented here support this mechanism, showing that beta-amyloid peptides (25-35), (1-39), and (1-42) activated the classical NADPH oxidase in rat primary culture of microglial cells and human phagocytes: 1) The exposure of the cells to beta-amyloid peptides stimulates the production of reactive oxygen intermediates; 2) the stimulation is associated with the assembly of the cytosolic components of NADPH oxidase on the plasma membrane, the process that corresponds to the activation of the enzyme; 3) neutrophils and monocytes of chronic granulomatous disease patients do not respond to beta-amyloid peptides with the stimulation of reactive oxygen intermediate production. Data are also presented that the activation of NADPH oxidase requires that beta-amyloid peptides be in fibrillary state, is inhibited by inhibitors of tyrosine kinases or phosphatidylinositol 3-kinase and by dibutyryl cyclic AMP, and is potentiated by interferon-gamma or tumor necrosis factor-alpha.     

Inhibition of -tyrosine-induced micronuclei production by phenylthiourea in human melanoma cells

It was previously found that -tyrosine oxidation product(s) are cytotoxic, genotoxic and increase the sister chromatid exchange (SCE) levels in human melanoma cells. In this work, the micronucleus assay has been performed on human melanotic and amelanotic melanoma cell lines (Carl-1 MEL and AMEL) in the presence of 1.0, 0.5 and 0.1 mM -tyrosine concentrations to investigate if melanin synthesis intermediate(s) increase micronuclei production. -Tyrosine oxidation product(s) increased the frequency of micronuclei in melanoma cells; 0.1 mM phenylthiourea (PTU), an inhibitor of -tyrosine oxidation by tyrosinase, lowered the micronucleus production to the control levels. The culture of melanoma cells with high -tyrosine in the culture medium resulted in a positive response to an ELISA-based apoptotic test. For comparison the effect of -tyrosine on micronuclei production in human amelanotic melanoma cells was also investigated; the micronucleus production in the presence of 1 mM -tyrosine in the culture medium was lower than that found with melanotic melanoma cells of the same cell line. The data suggest that melanin synthesis intermediates arising from -tyrosine oxidation may cause micronuclei production in Carl-1 human melanoma cells; the addition of PTU in the presence of -tyrosine decreased the frequency of micronuclei to about the control values thus the inhibition of melanogenesis may have some clinical implication in melanotic melanoma.     

Isolation and fractionation of gill cells from freshwater (Lasmigona costata) and seawater (Mesodesma mactroides) bivalves for use in toxicological studies with copper

Gills cells of the freshwater mussel Lasmigona costata and the seawater clam Mesodesma mactroides were isolated (mussel: chemical dissociation; clam: mechanical dissociation) and fractionated (Percoll gradient) into Fractions I and II. Mitochondrial dyes (DASPEI: mussel; MitoTracker^®: clam) and Na⁺, K⁺-ATPase activity measurement were used to distinguish between cells of Fractions I and II. For mussel and clam, 80.5 ± 1.5 and 48.3 ± 3.2 % of cells were in Fraction II, respectively. For both species, cells of Fraction II had higher fluorescence emission and higher enzyme activity than those of Fraction I, being characterized as ‘cells rich in mitochondria’. Cells of Fraction II were kept in saline solutions approximating the ionic composition of hemolymph either under control conditions (no Cu addition) or exposed (3 h) to copper (Cu: 5, 9 and 20 μg Cu/L). Cell viability and Cu and Na⁺ content were measured. For both species, Cu content was higher and Na⁺ content was lower in cells exposed to 20 μg Cu/L. Furthermore, a strong negative correlation was observed between cell Na⁺ and Cu content in the two bivalve species, indicating a possible competition between Cu and Na⁺ for ion-transporting mechanisms or binding sites at gill cells of Fraction II. Considering that Cu is an ionoregulatory toxicant in aquatic invertebrates, these preliminary toxicological data support the idea of using isolated gill cells rich in mitochondria to study the mechanisms underlying the acute toxicity of waterborne Cu in freshwater and marine bivalves.     

Isolation of adenylate cyclase mutants from Rhizobium meliloti deficient in nodulation

Six Rhizobium meliloti mutants were isolated after Tn5-mediated mutagenesis as resistant to inhibition by a mixture of amino acids (serine, methionine, glycine and leucine). All were defective in adenylate cyclase activity and failed to form nodules in infected roots of Medicago sativa. Furthermore, like other nodulation mutants, they showed altered motility and increased secretion of exopolysaccharides; addition of cAMP to the growth medium abolished some of these phenotypic defects. The possibility that adenylate cyclase participates in the transduction of signals inducing nodulation is discussed.     

Effect of heparin on proliferation and signalling in BC3H-1 muscle cells. Evidence for specific binding sites

We studied binding and growth inhibitory properties of different glycosaminoglycans in growing and differentiated BC3H-1 muscle cells. Heparin (10 micrograms/ml) and heparan sulfate (10 micrograms/ml) significantly inhibited DNA synthesis in growing and differentiated cells, as monitored by [3H]thymidine incorporation. Binding of heparin to BC3H-1 cells was specific and time-dependent. Heparan sulfate was the only glycosaminoglycan able to displace [3H]heparin (IC50, 3.2 x 10(-7) M), although it was 10-fold less effective than heparin itself (IC50, 3.6 x 10(-8) M). Scatchard analysis revealed the existence of high-affinity heparin binding sites (Kd, 5 x 10(-8) M). Furthermore, heparin inhibited serum-induced stimulation of inositol lipid turnover. Taken together, these results indicate that heparin inhibits BC3H-1 cell growth by interacting with the cell surface, possibly disrupting the flow of growth factor-related mitogenic signalling.     

Spectroscopic properties and related functions of the stigma measured in living cells of Euglena gracilis

The spectroscopic properties of stigma inside green and dark-grown cells and of isolated stigma globules have been studied by means of a microspectrophotometer built in the Laboratory. On the base of these results and of the analysis of the absorption spectra of a stigma suspension, cell suspension and cell methanolic extracts, it can be inferred that pigments localized in the stigma are free carotenoids which are not closely packed and do not show an ordered arrangement. Furthermore, the efficiency of the stigma as shading device is duscussed.