O1-conotoxin Tx6. 7 cloned from the genomic DNA of Conus textile that inhibits calcium currents

Background: Conotoxins exhibit great potential as neuropharmacology tools and therapeutic candidates due to their high affinity and specificity for ion channels, neurotransmitter receptors or transporters. The traditional methods to discover new conotoxins are peptide purification from the crude venom or gene amplification from the venom duct. Methods: In this study, a novel O1 superfamily conotoxin Tx6.7 was directly cloned from the genomic DNA of Conus textile using primers corresponding to the conserved intronic sequence and 3' UTR elements. The mature peptide of Tx6.7 (DCHERWDW CPASLLGVIYCCEGLICFIAFCI) was synthesized by solid-phase chemical synthesis and confirmed by mass spectrometry. Results: Patch clamp experiments on rat DRG neurons showed that Tx6.7 inhibited peak calcium currents by 59.29 ± 2.34% and peak potassium currents by 22.33 ± 7.81%. In addition, patch clamp on the ion channel subtypes showed that 10 µM Tx6.7 inhibited 56.61 ± 3.20% of the hCaV1.2 currents, 24.67 ± 0.91% of the hCaV2.2 currents and 7.30 ± 3.38% of the hNaV1.8 currents. Tx6.7 had no significant toxicity to ND7/23 cells and increased the pain threshold from 0.5 to 4 hours in the mouse hot plate assay. Conclusion: Our results suggested that direct cloning of conotoxin sequences from the genomic DNA of cone snails would be an alternative approach to obtaining novel conotoxins. Tx6.7 could be used as a probe tool for ion channel research or a therapeutic candidate for novel drug development.(AU)

Hydrochars produced with by-products from the sucroenergetic industry: a study of extractor solutions on nutrient and organic carbon release.

Hydrothermal carbonization transforms biomass into value-added material called hydrochar. The release of nutrients (P, N, Ca, Mg, and K) and organic carbon (TOC) from hydrochar in different extractive solutions was investigated in this study. Two sets of hydrochar were produced: (i) hydrochar prepared from sugarcane bagasse and vinasse mixture (BV-HC) and (ii) hydrochar prepared by the addition of H3PO4 to this mixture (BVA-HC). Both hydrochar types released significative amounts of nutrient and organic carbon, mainly Ca (5.0 mg g-1) in the mixture (KCl, K2SO4, NaOH, 1:1:1) extractive solution and TOC (72.6 mg g-1) in the NaOH extractive solution, for BV-HC. Nutrient release was influenced by pH and ionic strength. The release of P, Ca, and Mg was affected by the presence of insoluble phosphate phases in BVA-HC. The release of nutrients P, N, Ca, Mg, and K and organic carbon demonstrated that hydrochar has potential for soil application purposes.

Flow of essential elements in subcellular fractions during oxidative stress.

Essential trace elements are commonly found in altered concentrations in the brains of patients with neurodegenerative diseases. Many studies in trace metal determination and quantification are conducted in tissue, cell culture or whole brain. In the present investigation, we determined by ICP-MS Fe, Cu, Zn, Ca, Se, Co, Cr, Mg, and Mn in organelles (mitochondria, nuclei) and whole motor neuron cell cultured in vitro. We performed experiments using two ways to access oxidative stress: cell treatments with H2O2 or Aß-42 peptide in its oligomeric form. Both treatments caused accumulation of markers of oxidative stress, such as oxidized proteins and lipids, and alteration in DNA. Regarding trace elements, cells treated with H2O2 showed higher levels of Zn and lower levels of Ca in nuclei when compared to control cells with no oxidative treatments. On the other hand, cells treated with Aß-42 peptide in its oligomeric form showed higher levels of Mg, Ca, Fe and Zn in nuclei when compared to control cells. These differences showed that metal flux in cell organelles during an intrinsic external oxidative condition (H2O2 treatment) are different from an intrinsic external neurodegenerative treatment.

Sulfate removal from waste chemicals by precipitation.

Chemical oxidation using Fenton's reagent has proven to be a viable alternative to the oxidative destruction of organic pollutants in mixed waste chemicals, but the sulfate concentration in the treated liquor was still above the acceptable limits for effluent discharge. In this paper, the feasibility of sulfate removal from complex laboratory wastewaters using barium and calcium precipitation was investigated. The process was applied to different wastewater cases (two composite samples generated in different periods) in order to study the effect of the wastewater composition on the sulfate precipitation. The experiments were performed with raw and oxidized wastewater samples, and carried out according to the following steps: (1) evaluate the pH effect upon sulfate precipitation on raw wastewaters at pH range of 2-8; (2) conduct sulfate precipitation experiments on raw and oxidized wastewaters; and (3) characterize the precipitate yielded. At a concentration of 80 g L(-1), barium precipitation achieved a sulfate removal up to 61.4% while calcium precipitation provided over 99% sulfate removal in raw and oxidized wastewaters and for both samples. Calcium precipitation was chosen to be performed after Fenton's oxidation; hence this process configuration favors the production of higher quality precipitates. The results showed that, when dried at 105 degrees C, the precipitate is composed of hemidrate and anhydrous calcium sulfate ( approximately 99.8%) and trace metals ( approximately 0.2%: Fe, Cr, Mn, Co, Ag, Mg, K, Na), what makes it suitable for reuse in innumerous processes.

[Zinc, copper, iron, calcium, phosphorus and magnesium content of maternal milk during the first 3 weeks of lactation]. / Contenido de zinc, cobre, hierro, calcio, fósforo y magnesio en leche materna en los primeros días de lactación.

Zinc, Copper, Iron. Calcium Phosphorous and Magnesium contents were determined in early milk samples in 72 mothers from Caracas city. The samples were collected during three different lactation stages: calostro (3 days), transitional (7 days) and mature milk (21 days). The more significant changes in the concentration of the studied elements were observed during the first two weeks, them they stabilize during the third week. The Zn, Cu, Fe, Ca, P and Mg average concentration found in calostro samples were 7.1 +/- 2.5 micrograms/ml; 0.52 +/- 0.15 microgram/ml; 0.49 +/- 0.14 microgram/ml; 214 +/- 62 micrograms/ml, 107 +/- 27 micrograms/ml and 33.3 +/- 7.5 micrograms/ml. respectively. For the transitional milk samples the average concentration found for the studied elements were: 4.0 +/- 1.0 micrograms/ml; 0.50 +/- 0.10 microgram/ml; 0.38 +/- 0.08 microgram/ml, 292 +/- 62 micrograms/ml; 213 +/- 36 micrograms/ml and 30.4 +/- 5.2 micrograms/ml. For the mature milk samples the results were: 2.8 +/- 2.7 micrograms/ml; 0.47 +/- 0.08 microgram/ml; 0.36 +/- 0.09 microgram/ml; 244 +/- 49 micrograms/ml; 175 +/-35 micrograms/ml and 25.2 +/- 3.3 micrograms/ml. The concentration range for all trace elements studied (Cu, Fe and Zn) can be considered normal. For the major elements (Ca, P and Mg) the results obtained in our work are similar to those reported for other countries. These facts allows to conclude that the nutritional state of this mother population is adequate to satisfy the lactate's requirements during their first live stage.

Minerals content of Paraguayan yerba mate (Ilex paraguariensis, S.H.).

Minerals content (Fe, Ca, Mn, Mg, Na, K, Zn and Cu) of the leaves of Paraguayan yerba mate (Ilex paraguariensis, S.H.) as well as of commercial products has been determined by atomic absorption spectrophotometry. Considerable amounts of iron and calcium and remarkably high content of magnesium and potassium have been found both in the leaves and in the commercial products. Highly significant difference between the Fe content of the leaves and that of the commercial products has been found. Also, highly significant difference has been found between the Fe and Ca content of the higher quality-greater sale reputed commercial products and that of the lower quality-smaller sale reputed ones. Seasonal and soil linked variations are also presented. The nutritional and quality control significance of these findings for the MERCOSUR (Southern South America Economical Community) are discussed.