The endogenous cannabimimetic substance, anandamide (
N-arachidonoyl-ethanolamine) and the recently isolated sleep-inducing factor, oleoyl-amide (
cis-9,10-octadecenoamide), belong to two neuroactive ...fatty acid amide classes whose action in mammals has been shown to be controlled by enzymatic amide bond hydrolysis. Here we report the partial characterisation and purification of ‘anandamide amidohydrolase’ from membrane fractions of N18 neuroblastoma cells, and provide evidence for a further and previously unsuspected role of this enzyme. An enzymatic activity catalysing the hydrolysis of
14Canandamide was found in both microsomal and 10,000 ×
g pellet fractions. The latter fractions, which displayed the highest
V
max for anandamide, were used for further characterisation of the enzyme, and were found to catalyse the hydrolysis also of
14Coleoyl-amide, with an apparent
K
m of 9.0 ± 2.2
μM.
14Canandamide- and
14Coleoyl-amide-hydrolysing activities: (i) exhibited identical pH- and temperature-dependency profiles; (ii) were inhibited by alkylating agents; (iii) were competitively inhibited by the phospholipase A
2 inhibitor arachidonyl-trifluoromethyl-ketone with the same IC
50 (3 μM); (iv) were competitively inhibited by both anandamide (or other polyunsaturated fatty acid-ethanolamides) and oleoyl-amide. Proteins solubilised from 10,000 ×
g pellets were directly analysed by isoelectric focusing, yielding purified fractions capable of catalysing the hydrolysis of both
14Canandamide and
14Coleoyl-amide. These data suggest that ‘anandamide amidohydrolase’ enzymes, such as that characterised in this study, may be used by neuronal cells also to hydrolyse the novel sleep-inducing factor oleoyl-amide.
Aspartate aminotransferase from the archaebacterium Sulfolobus solfataricus, a thermoacidophilic organism isolated from an acidic hot spring (optimal growth conditions: 87 degrees C, pH 3.5) was ...purified to homogeneity. The enzyme is a dimer (Mr subunit = 53,000) showing microheterogeneity when submitted to chromatofocusing and/or isoelectric focusing analysis (two main bands having pI = 6.8 and 6.3 were observed). The N-terminal sequence (22 residues) does not show any homology with any stretch of known sequence of aspartate aminotransferases from animal and bacterial sources. The apoenzyme can be reconstituted with pyridoxamine 5′-phosphate and/or pyridoxal 5′-phosphate, each subunit binding 1 mol of coenzyme. The absorption maxima of the pyridoxamine and pyridoxal form are centered at 325 and 335 nm, respectively; the shape of the pyridoxal form band does not change with pH. The enzyme has an optimum temperature higher than 95 degrees C, and at 100 degrees C shows a half-inactivation time of 2 h. The above properties seem to be unique even for enzymes from extreme thermophiles (Daniel, R. M. (1986) in Protein Structure, Folding, and Design (Oxender, D. L., ed) pp. 291-296, Alan R. Liss, Inc., New York) and lead to the conclusion that aspartate aminotransferase from S. solfataricus is one of the most thermophilic and thermostable enzymes so far known.
Amifostine (WR‐2721; Ethyol) is a well‐known cytoprotector, but a possible role in preventing extrahaematological toxicity after high‐dose therapy (HDT) has never been investigated. We compared two ...historical groups of patients who either received (group A, n = 35) or did not receive (group B, n = 33) amifostine (740 mg/m2) before high‐dose (HD) melphalan, followed by autologous infusion of peripheral blood progenitor cells (PBPCs). Amifostine was well tolerated at this dose level. Emesis grade 1–2 was the most important side‐effect, but the interruption of infusion was never required. The incidence and median duration of severe mucositis (grade 3–4) was 21% and 0 d (range 0–11 d) in group A and 53% and 7 d (range 0–11 d) in group B. The duration of analgesic therapy was also significantly lower in group A (0 d; range 0–12) than in group B (6 d, range 0–20) (P = 0·0001). Severe diarrhoea (3% vs. 25%; P = 0·01) and emesis (9% vs. 34%; P = 0·01) were also reduced in group A in comparison with group B. No differences were observed between the two groups for haematological recovery. This retrospective study strongly suggests that amifostine can reduce severe mucositis and the use of analgesic drugs in this setting. A randomized study is warranted to confirm these preliminary results.
A novel diketopiperazine, named cyclo-(D-pipecolinyl-L-isoleucine) (DKP 1), and 7 known diketopiperazines were isolated from the cell-free culture supernatant of the Antarctic psychrophilic bacterium ...Pseudoalteromonas haloplanktis TAC125. Two diketopiperazines containing pipecolinyl moiety were isolated for the first time from a natural source. Two new linear peptides, stable to bacterial proteolytic enzymes, were also characterized. The structures of the isolated compounds were elucidated by means of spectroscopic data (1D-, 2D-NMR, EIMS, FABMS, and ESIMS/MS) and chiral high-performance liquid chromatography. The potential antioxidant activity of the isolated compounds was evaluated by a DPPH free radical scavenging assay.
Possible risk factors for psoriasis were studied by means of a retrospective evaluation of epidemiological and clinical features in 472 clinical records of outpatients affected by plaques psoriasis. ...We calculated the relation between the categorical variables with chi-squares contingency table analysis (with Yates correction) and odds ratio (O.R.) from the two variables with their respective confidence intervals at 95%. Among patients 267 were men (57%) and 205 women (43%); the mean age was 46.5+/-16.7 years, with no differences in sex. A significant relationship was found between age at onset < or =40 years and familiarity for psoriasis (p<0.0001-O.R.=2.71, C.I..95%: 2.12-3.48). The percentage of men with extensive lesions (extension >30%) was significantly higher than of women (p<0.05-O.R.=1.60, C.I..95%:1.35-1.90). Moreover, a significant association between extension of lesions and BMI was detected (p<0.0107-O.R.=2.10, C.I.95%:1.33-3.30) and the strength of this relation was assessed using the linear regression test. Mean age at onset was significantly lower in patients systemically treated (p<0.0003-O.R.=2.61, C.I.95%: 1.69-4.04). The results obtained can provide the basis for a standard clinical record, including data for a prospective study that may confirm the relation between psoriasis and some risk factors.
Post-surgical hemifrontal anhidrosis Fernández Rodríguez, J; Cariñena Amigo, A; Sevillano Torrado, C ...
Revista española de anestesiología y reanimación
61, Številka:
10
Journal Article
A cytogenetic analysis was carried out on specimens of Epinephelus marginatus (Lowe, 1834) from three localities in the Mediterranean Sea, to deepen knowledge of the chromosome complement of the ...species and identify any possible populatgion-specific cytogenetic markers. All specimens had a 2n = 48 acrocentric karyotype with two Ag-, chromomycin A(3)- and C-positive NORS (nucleolar organizer regions) in the subcentromeric region of the smallest chromosome pair.
We tested the hypothesis that controlled reperfusion with leukocyte-depleted blood could improve myocardial protection by reducing the oxidative stress in patients undergoing myocardial ...revascularization. Thirty-four patients receiving antegrade/retrograde blood cardioplegia were divided into: group A: 11 patients with ejection fractions (EF) less than 35%, treated with leukocyte-depleted controlled blood reperfusion, group B: 11 patients with EF less than 35% in whom no leukocyte depletion was performed, group C: 6 patients with EF more than 45% treated as group A and group D: 6 patients with EF more than 45% without leukocyte depletion. To asses the oxidative stress, we evaluated total, total oxidized (GSSX), and reduced glutathione (GSH) in coronary sinus plasma, immediately before cross-clamping the aorta (T0), and at 0 (T1), 15 (T2) and 30 (T3) min after unclamping it. In groups A and B a significant shift towards oxidation of redox status of glutathione (GSH/GSSX) at T1 vs T0 was observed. Glutathione redox ratio remained low in group B while in group A it returned to the basal value at T2 with a significant difference from group B at T2 and T3. No differences were observed between groups C and D. In conclusion, our data show that leukocyte-depleted reperfusion can afford a better myocardial protection in patients with left ventricular dysfunction, while it seems unnecessary in patients with normal EF.
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