Conicamin, a novel histamine antagonist from the mediterranean tunicate Aplidium conicum

In addition to the known 6-bromo-hypaphorine (2) and plakohypaphorine-A (3), the methanol extract of the Mediterranean tunicate Aplidium conicum was shown to contain conicamin, a novel indole alkaloid having histamine-antagonistic activity which structure was determined to be 1 on the basis of the spectral data.     

Amoxicillin/tetracycline combinations are inadequate as alternative therapies for Helicobacter pylori infection

Background. Triple therapy with proton pump inhibitors or ranitidine bismuth citrate, clarithromycin and either amoxicillin or nitroimidazole derivatives are the present gold standards for cure of Helicobacter pylori infection. However, primary resistance to either clarithromycin or nitroimidazole derivatives is increasing and alternative therapies are needed. Aim. To determine the efficacy and safety of three regimens consisting of amoxicillin and tetracycline or doxycycline combined with either lansoprazole or ranitidine bismuth citrate. Methods. Two hundred and seventy H. pylori infected patients were randomly given one of the following treatments: amoxicillin 1 g twice a day (b.i.d.) plus tetracycline 500 mg four times a day (q.i.d.) with either lansoprazole 30 mg b.i.d. (group LAT) or ranitidine bismuth citrate 400 mg b.i.d. (group RBCAT) for 7 days and amoxicillin 1 g b.i.d. plus doxycycline 100 mg b.i.d. and lansoprazole 30 mg b.i.d. for 14 days (group LAD). Eradication rate was assessed by UBT at 4–6 weeks after therapy. Results. The three groups (LAT, RBCAT, and LAD) of patients achieved eradication rates of 35% (25–45), 20% (12–29) and 36% (25–46), respectively, on intention‐to‐treat analysis. Patient compliance was optimal and side‐effects minimal in all three groups. Conclusions. Although the amoxicillin/tetracycline combination is attractive (inexpensive, safe, and with low primary resistance rate), it can not be recommended for H. pylori eradication.     

Seed storage conditions change the germination pattern of clonal growth plants in Mediterranean salt marshes

The effect of salinity level and extended exposure to different salinity and flooding conditions on germination patterns of three salt-marsh clonal growth plants (Juncus subulatus, Scirpus litoralis, and S. maritimus) was studied. Seed exposure to extended flooding and saline conditions significantly affected the outcome of the germination process in a different, though predictable, way for each species, after favorable conditions for germination were restored. Tolerance of the germination process was related to the average salinity level measured during the growth/germination season at sites where established individuals of each species dominated the species cover. No relationship was found between salinity tolerance of the germination process and seed response to extended exposure to flooding and salinity conditions. The salinity response was significantly related to the conditions prevailing in the habitats of the respective species during the unfavorable (nongrowth/nongermination) season. Our results indicate that changes in salinity and hydrology while seeds are dormant affect the outcome of the seed-bank response, even when conditions at germination are identical. Because these environmental-history-dependent responses differentially affect seed germination, seedling density, and probably sexual recruitment in the studied and related species, these influences should be considered for wetland restoration and management.     

Feeding level in the period previous to the late fattening phase influences fat composition at slaughter in free-ranged Iberian pigs

A group of 11 pigs was fed with 70 g feed per kg of metabolic weight (H pigs) and another group of 11 pigs was fed with 50g feed per kg of metabolic weight (L pigs). In both experimental groups (H and L pigs), it was observed that the higher initial proportion of C16:0, C18:0 and C18:2 (n-6) in backfat at the beginning of the free-range feeding period, the greater decrease rate of these fatty acid proportions regarding weight gain during the free-range fattening period took place. On the other hand, the greater initial proportion of C18:1 (n-9), the smaller increase rate in the concentration of this fatty acid was observed. The intramuscular neutral lipids from L pigs had higher C18:3 (n-3) and lower proportions of monounsaturated fatty acids (MUFA) than those from H pigs, while intramuscular polar lipids from L pigs had significantly higher proportions of C18:0, PUFA, C18:3 (n-3) and (n-3) and significantly lower MUFA and C18:1 (n-9) proportions than those from H pigs. The alpha-tocopherol concentration found in Longissimus dorsi from L pigs was significantly higher (p < 0.012) than those from H pigs.     

Scale-up of virus-like particles production: effects of sparging, agitation and bioreactor scale on cell growth, infection kinetics and productivity

The baculovirus-insect cells expression system was used for the production of self-forming Porcine parvovirus (PPV) like particles (virus-like particles, VLPs) in serum-free medium. At 2l bioreactor scale an efficient production was achieved by infecting the culture at a concentration of 1.5 x 10(6)cells/ml using a low multiplicity of infection of 0.05 pfu per cell. In a continuous bioreactor, it was shown that the uninfected insect cells were not sensitive to local shear stress values up to 2.25 N/m2 at high Reynolds numbers (1.5 x 10(4)) in sparging conditions. Uninfected insect cells can be grown at scaled-up bioreactor at high agitation and sparging rates as long as vortex formation is avoided and bubble entrapment is minimized. An efficient process scale-up to 25 l bioreactor was made using constant shear stress criteria for scale-up. The kinetics of baculovirus infection at low multiplicity of infection, either at different cell concentration or at different scales, are very reproducible, despite the different turbulence conditions present in the bioreactor milieu. The results suggest that the infection kinetics is controlled by the rate of baculovirus-cell receptor attachment and is independent of the bioreactor hydrodynamic conditions. Furthermore, the achieved specific and volumetric productivities were higher at the 25 l scale when compared to the smaller scale bioreactor. Different rates of cell lysis after infection were observed and seem to fully explain both the shift in optimal harvest time and the increase in cell specific productivity. The results emphasize the importance of integrated strategies and engineering concepts in process development at bioreactor stage with the baculovirus insect cell system.     

Modified mattress suture technique to correct anterior septal deviation

The purpose of this study was to evaluate the efficacy of a modified mattress suture technique in septal anterior deviation correction and to compare it with the Hinderer technique. This was a randomized, controlled, single-blind clinical trial. Fifty patients with anterior septal deviation were assigned to one of two surgery treatment groups: in group A, 25 patients underwent anterior septoplasty with modified mattress suture technique; and in group B, 25 patients underwent anterior septoplasty with the Hinderer technique. Subjective (oral respiration, epistaxis, rhinorrhea, and nasal obstruction) and objective (anterior rhinomanometry) parameters were analyzed and compared between groups. Better results in obstructive deflections were achieved with the modified mattress suture technique. Unilateral and total nasal resistances improved during the 6-month follow-up in patients who underwent anterior septoplasty with the modified mattress suture technique, and there was a significant difference in the values when compared with patients who underwent anterior septoplasty with rhinoplasty using the Hinderer technique (p = 0.003). The modified mattress suture technique had better subjective and objective results than rhinoplasty with the Hinderer technique and therefore could be considered as an alternative technique in anterior septal surgery.     

Inhibitory effect of ginger (Zingiber officinale) on rat ileal motility in vitro

Ginger (Zingiber officinale rhizome) is a widespread herbal medicine mainly used for the treatment of gastrointestinal diseases, including dyspepsia, nausea and diarrhoea. In the present study we evaluated the effect of this herbal remedy on the contractions induced by electrical stimulation (EFS) or acetylcholine in the isolated rat ileum. Ginger (0.01-1000 microg/ml) inhibited both EFS- and acetylcholine-evoked contractions, being more potent in inhibiting the contractions induced by EFS. The depressant effect of ginger on EFS-induced contractions was reduced by the vanilloid receptor antagonist capsazepine (10(-5) M), but unaffected by the alpha(2)-adrenergic antagonist yohimbine (10(-7) M), the CB(1) receptor antagonist SR141716A (10(-6) M), the opioid antagonist naloxone (10(-6) M) or by the NO synthase inhibitor L-NAME (3 x 10(-4) M). Zingerone (up to 3 x 10(-4) M), one of the active ingredients of ginger, did not possess inhibitory effects. It is concluded that ginger possesses both prejunctional and postjunctional inhibitory effects on ileal contractility; the prejunctional inhibitory effect of ginger on enteric excitatory transmission could involve a capsazepine-sensible site (possibly vanilloid receptors).     

Copper, iron, and Zinc contents in human milk during the first three months of lactation

The aim of the study has been to analyze the evolution of copper, iron, and zinc contents in human milk, from colostrum to the third postpartum month, following a longitudinal design, under specific conditions of sample collection and to apply an analytical procedure previously optimized to reduce any variation outside physiological lactation. The copper, iron, and zinc concentrations in 144 milk samples from 39 healthy puerpera women, were analyzed in five stages by flame atomic absorption spectrometry, following a standardized protocol. Copper presented a gradual decrease from 0.38 mg/L to 0.19 mg/L by the 90th day; the particular analysis from colostrum to transitional milk manifested the following two tendencies. Whereas an increase from 0.19 to 0.42 mg/L was observed in some women, a decrease from 0.53 to 0.45 mg/L was detected in others; therefore, copper presented two significant behaviors in the evolution from colostrum to transitional milk. In both cases, the evaluated changes were significant. The iron content varied from 0.56 to 0.40 mg/L by the 30th day, remaining constant until the first trimester concluded. The average zinc concentration decreased sharply from 7.99 to 3.3 mg/L on d 15; the rate of decrease slowed down gradually until 1.05 mg/L.     

Formation of an RNase A derivative containing an aminosuccinyl residue in place of asparagine 67

At acidic pH, Asp67 and beta-Asp67 (beta-Asp: isoaspartic acid residue) derivatives of RNase A, obtained by selective deamidation of the parent enzyme, spontaneously produces a new derivative containing an aminosuccinyl residue (Asu). The overall secondary structure of the protein chain does not change as a consequence of this substitution, while the catalytic activity on RNA is reduced to about 25%. The pH dependence of the first-order rate constants for the Asu formation has a bell-shaped profile, the maximum being close to the pK(a) of the aspartic acid side chains. Moreover, the values of the rate constants are of the same magnitude of those measured for Asp-containing peptides whose sequence mimics the Asu formation site of the enzyme. This feature indicates that Asp67 and beta-Asp67 residues in the deamidated RNase A derivatives are sited in a region flexible enough to permit the cyclization of the carboxylic side chain to succinimide ring. These results are discussed at the light on to the three-dimensional structure and the thermodynamic stability of the aspartic acid derivatives of RNase A.     

Zinc transport and metallothionein secretion in the intestinal human cell line Caco-2

Caco-2, a human cell line, displays several biochemical and morphological characteristics of differentiated enterocytes. Among these is the ability to transport zinc from the apical to the basal compartment. This process was enhanced following exposure by the apical compartment to increasing concentrations of the metal. High pressure liquid chromatography fractionation of the media obtained from cells labeled with radioactive zinc showed that metallothioneins (MTs), small metal-binding, cysteine-rich proteins), were present in the apical and basal media of controls as well as in cells grown in the presence of high concentrations of zinc. Following exposure to the metal, the levels of Zn-MTs in the apical medium increased, while in the basal compartment the greatest part of zinc appeared in a free form with minor changes in the levels of basal MTs. Metabolic labeling experiments with radioactive cysteine confirmed the apical secretion of MTs. A stable transfectant clone of Caco-2 cells (CL11) was selected for its ability to express constitutively high levels of the mouse metallothionein I protein. This cell line showed an enhanced transport of the metal following exposure to high concentrations of zinc and a constitutive secretion of the mouse metallothionein I protein in the apical compartment. Together, these findings strongly support the hypothesis of a functional role between the biosynthesis and secretion of MTs and the transport of zinc in intestinal cells.