Riconoscimento e Dosamento della Psilocibina

It is possible to detect and dose Psylocibine, a psychoneurotropic substance occurring inPsilocybe, Stropharia, Panaeolus etc., with a reaction already used to detect Tryptophane and indole-auxins (6) (7) (8) (9). This method is more sensitive than U.V. measurement and Keller's reaction (4) (see fig. 3). Optical density is proportional to concentration from 0,005 to 0,050µmoles per ml when measured in a 1 cm cell in the Beckman spectrophotometer. Tryptophane, indole-auxins and 2-deoxy-sugars could interfere (6) (7) (8) (9) (10). procedure: In a test tube are pipetted:

1. 1 % solution of fructose in H₂O : ml 0.1
2. 5 % solution of cysteine hydrochloride : ml 0.2
3. methanolic solution to be tested : ml 1.0
4. conc. H₂SO₄/water 70/30 (v.v.) : ml 3.0 the mixture is allowed to cool off at room temperature, a pink color develops and reaches its maximum of intensity in about 90 minutes.

The absorption spectrum of the chromophore exhibits a maximum at 512 mµ (see fig. 2). The ratio E₅₁₂ in the reaction mixture/E₂₆₇ in U.V. is about 1,9 and differs from other indole-derivatives. (see Tab. II). The reaction can also be employed to detect and dose Psylocibine-spots in paper and thin-layer chromatography spraying with the following mixture prepared just before use: fructose g 1.0 cysteine hydrochloride g 1.0 3 N H₂SO₄ ml 100 Paper — chromatograms are dried in an oven (70°C) for 5 minutes, chromatoplates for 10–15 minutes. Psylocibine appears as a pink spot.     

Detection of Tissue Factor Antigen and Coagulation Activity in Coronary Artery Thrombi Isolated from Patients with ST-Segment Elevation Acute Myocardial Infarction

Introduction

Although ruptured atherosclerotic plaques have been extensively analyzed, the composition of thrombi causing arterial occlusion in patients with ST-segment elevation acute myocardial infarction has been less thoroughly investigated. We sought to investigate whether coagulant active tissue factor can be retrieved in thrombi of patients with STEMI undergoing primary percutaneous coronary intervention.

Methods

Nineteen patients with ST-segment elevation acute myocardial infarction referred for primary percutaneous coronary intervention were enrolled in this study. Coronary thrombi aspirated from coronary arteries were routinely processed for paraffin embedding and histological evaluation (4 patients) or immediately snap frozen for evaluation of tissue factor activity using a modified aPTT test (15 patients). Immunoprecipitation followed by immunoblotting was also performed in 12 patients.

Results

Thrombi aspirated from coronary arteries showed large and irregular areas of tissue factor staining within platelet aggregates, and in close contact with inflammatory cells. Some platelet aggregates stained positive for tissue factor, whereas others did not. Monocytes consistently stained strongly for tissue factor, neutrophils had a more variable and irregular tissue factor staining, and red blood cells did not demonstrate staining for tissue factor. Median clotting time of plasma samples containing homogenized thrombi incubated with a monoclonal antibody that specifically inhibits tissue factor-mediated coagulation activity (mAb 5G9) were significantly longer than their respective controls (88.9 seconds versus 76.5 seconds, respectively; p<0.001). Tissue factor was also identified by immunoprecipitation in 10 patients, with significant variability among band intensities.

Conclusions

Active tissue factor is present in coronary artery thrombi of patients with ST-segment elevation acute myocardial infarction, suggesting that it contributes to activate the coagulation cascade ensuing in coronary thrombosis.     

Do mycoplasmas cause human cancer?

A linkage between mycoplasmas and malignancy was mainly proposed in the 1960s when human-associated mycoplasmas were becoming of interest given the novel characterization of the human respiratory pathogen Mycoplasma pneumoniae. Associations with leukemia and other malignancies, however, were largely ascribed to tissue-culture contamination, which is now recognized as a significant potential problem in molecular biology circles. A few epidemiological studies, however, continue to raise concern over such a linkage. As well, in vitro data have demonstrated the potential for some mycoplasmas to induce karyotypic changes and malignant transformation during chronic tissue-culture infestation. As cellular and molecular mechanisms for such transformation become studied, a resurgence of interest in this area is inevitable. A role for mycoplasmas in malignancy of any sort is conjectural, but there remains a need to continue with focussed epidemiological and laboratory investigations.     

Miocene ostracodes of cold seep settings from northern Apennines (Italy)

The Miocene Termina Formation of the northern Apennines (Italy) contains thin marly limestone or marly sandstone bodies rich in macrofauna mainly consisting of large lucinid clams. Modern representatives of this fauna occur in cold seep settings where they house chemosymbiotic bacteria. Moreover, these rocks record a δ¹³C-depletion confirming their origin influenced by a cold seep. The Miocene sections of Sasso delle Streghe and Sarsetta (near Modena) represent classic examples of cold seeps. These sections consist of marls and marly sandstones, respectively, with lucinids (Sasso delle Streghe) or bioclastic sands with lucinids (Sarsetta); both sections are capped by the Sandstones of Montebaranzone. They yield ostracodes that are mainly represented by deep-water filter-feeder species (Platycopa, i.e. Cytherella sp.) and numerous deposit-feeders (Podocopa: i.e. Neonesidea and Krithe), together with species frequently occurring in shallow water environments. These assemblages are able to colonize disaerobic environments such as cold seeps. Although some authors consider the filter-feeders as dominant taxa in disaerobic settings, our data do not provide evidence of significant differences in the ostracode assemblage composition between the deposits originated in seafloors with or without seepage influence. However, the number of specimens is greater in seafloors devoid of seepage influence.     

Synthesis and antibacterial activity evaluation of a novel cotton fiber (Gossypium barbadense) ampicillin derivative

We prepared cellulose cotton fibers containing ampicillin moieties and evaluated their antibacterial activity. In spite of recent progress in experimental and clinical medicine, the problem of chronic wounds treatment remains to be solved. In fact conventional methods are based on solutions of antibiotics and antiseptics and ointment bandages but the efficacy of this method is low and so the idea to use modified cotton gauzes would have to prevent infections insorgence during wounds healing. Ampicillin, a large spectrum antibiotic, was covalently coupled to cellulose backbone of hydrophilic cotton fibers by a heterogeneous synthesis to produce a functionalized biopolymer with a satisfactory degree of substitution (DS) and antibacterial activity. The obtained biopolymer was characterized by infrared spectroscopy (FT-IR). Finally, the antibacterial activity in inhibiting microorganism growth in Petri dishes, was evaluated. The results suggested that these biomaterials posses an excellent “in vitro” antibacterial activity and so they can be efficiently employed in biomedical fields for chronic wounds management to ensure a valid protection against infections and contaminations. Biopolymers so functionalized were found to be very efficient to contrast sensible bacteria growth.     

Circulating miR-184 is a potential predictive biomarker of cardiac damage in Anderson–Fabry disease

Enzyme replacement therapy (ERT) is a mainstay of treatment for Anderson–Fabry disease (AFD), a pathology with negative effects on the heart and kidneys. However, no reliable biomarkers are available to monitor its efficacy. Therefore, we tested a panel of four microRNAs linked with cardiac and renal damage in order to identify a novel biomarker associated with AFD and modulated by ERT. To this end, 60 patients with a definite diagnosis of AFD and on chronic ERT, and 29 age- and sex-matched healthy individuals, were enrolled by two Italian university hospitals. Only miR-184 met both conditions: its level discriminated untreated AFD patients from healthy individuals (c-statistic = 0.7522), and it was upregulated upon ERT (P < 0.001). On multivariable analysis, miR-184 was independently and inversely associated with a higher risk of cardiac damage (odds ratio = 0.86; 95% confidence interval [CI] = 0.76–0.98; P = 0.026). Adding miR-184 to a comprehensive clinical model improved the prediction of cardiac damage in terms of global model fit, calibration, discrimination, and classification accuracy (continuous net reclassification improvement = 0.917, P < 0.001; integrated discrimination improvement [IDI] = 0.105, P = 0.017; relative IDI = 0.221, 95% CI = 0.002–0.356). Thus, miR-184 is a circulating biomarker of AFD that changes after ERT. Assessment of its level in plasma could be clinically valuable in improving the prediction of cardiac damage in AFD patients.Subject terms: Prognostic markers, Cardiovascular diseases     

Myocardial Mitochondrial and Contractile Function Are Preserved in Mice Lacking Adiponectin

Adiponectin deficiency leads to increased myocardial infarct size following ischemia reperfusion and to exaggerated cardiac hypertrophy following pressure overload, entities that are causally linked to mitochondrial dysfunction. In skeletal muscle, lack of adiponectin results in impaired mitochondrial function. Thus, it was our objective to investigate whether adiponectin deficiency impairs mitochondrial energetics in the heart. At 8 weeks of age, heart weight-to-body weight ratios were not different between adiponectin knockout (ADQ^-/-) mice and wildtypes (WT). In isolated working hearts, cardiac output, aortic developed pressure and cardiac power were preserved in ADQ^-/- mice. Rates of fatty acid oxidation, glucose oxidation and glycolysis were unchanged between groups. While myocardial oxygen consumption was slightly reduced (-24%) in ADQ^-/- mice in isolated working hearts, rates of maximal ADP-stimulated mitochondrial oxygen consumption and ATP synthesis in saponin-permeabilized cardiac fibers were preserved in ADQ^-/- mice with glutamate, pyruvate or palmitoyl-carnitine as a substrate. In addition, enzymatic activity of respiratory complexes I and II was unchanged between groups. Phosphorylation of AMP-activated protein kinase and SIRT1 activity were not decreased, expression and acetylation of PGC-1α were unchanged, and mitochondrial content of OXPHOS subunits was not decreased in ADQ^-/- mice. Finally, increasing energy demands due to prolonged subcutaneous infusion of isoproterenol did not differentially affect cardiac contractility or mitochondrial function in ADQ^-/- mice compared to WT. Thus, mitochondrial and contractile function are preserved in hearts of mice lacking adiponectin, suggesting that adiponectin may be expendable in the regulation of mitochondrial energetics and contractile function in the heart under non-pathological conditions.     

Docosahexaenoic acid induces apoptosis in lung cancer cells by increasing MKP-1 and down-regulating p-ERK1/2 and p-p38 expression

Different agents able to modulate apoptosis have been shown to modify the expression of the MAP-kinase-phosphatase-1 (MKP-1). The expression of this phosphatase has been considered a potential positive prognostic factor in lung cancer, and smoke was shown to reduce the levels of MKP-1 in ferret lung. Our aim was to assess whether the n-3 polyunsaturated fatty acid docosahexaenoic acid (DHA), known to inhibit the growth of several cancer cells mainly inducing apoptosis, may exert pro-apoptotic effect in lung cancer cells by modifying MKP-1 expression. We observed that DHA increased MKP-1 protein and mRNA expression and induced apoptosis in different lung cancer cell lines (mink Mv1Lu adenocarcinoma cells, human A549 adenocarcinoma and human BEN squamous carcinoma cells). We inhibited the pro-apoptotic effect of DHA by treating the cells with the phosphatase inhibitor Na₃VO₄ or by silencing the MKP-1 gene with the specific siRNA. This finding demonstrated that the induction of apoptosis by DHA involved a phosphatase activity, specifically that of MKP-1. DHA reduced also the levels of the phosphorylated MAP-kinases, especially ERK1/2 and p38. Such an effect was not observed when the MKP-1 gene was silenced. Altogether, the data provide evidence that the DHA-induced overexpression of MKP-1 and the resulting decrease of MAP-kinase phosphorylation by DHA may underlie the pro-apoptotic effect of this fatty acid in lung cancer cells. Moreover, they support the hypothesis that DHA may exert chemopreventive action in lung cancer.     

The oxidative stress and the mitochondrial dysfunction caused by endotoxemia are prevented by α-lipoic acid

The aims of this work were to study the mitochondrial function and to evaluate (a) the oxidative stress in real time in an acute model of endotoxemia and (b) the effect of α-lipoic acid (LA, 100 mg/kg) as a therapeutic strategy to be considered. In rats treated with lipopolisaccharide (LPS, 10 mg/kg), a 1.4-fold increase was observed in in situ skeletal muscle chemiluminescence. Experimental sepsis increased oxygen consumption in tissue cubes (1 mm³) by 30% for heart and diaphragm and impaired state 3 mitochondrial respiration rate in the three organs (liver, diaphragm and heart) studied. Only complex I activity in heart and diaphragm and complex IV activity in diaphragm were found impaired in this septic model. The production of NO by submitochondrial membranes was found increased by 80% in the diaphragm and by 35% in the heart of septic rats. The treatment with LA prevented the oxidative stress and mitochondrial dysfunction observed in this model.