Dynamics of fine root carbon in Amazonian tropical ecosystems and the contribution of roots to soil respiration

Radiocarbon (¹⁴C) provides a measure of the mean age of carbon (C) in roots, or the time elapsed since the C making up root tissues was fixed from the atmosphere. Radiocarbon signatures of live and dead fine (<2 mm diameter) roots in two mature Amazon tropical forests are consistent with average ages of 4–11 years (ranging from <1 to >40 years). Measurements of ¹⁴C in the structural tissues of roots known to have grown during 2002 demonstrate that new roots are constructed from recent (<2‐year‐old) photosynthetic products. High Δ¹⁴C values in live roots most likely indicate the mean lifetime of the root rather than the isotopic signature of inherited C or C taken up from the soil. Estimates of the mean residence time of C in forest fine roots (inventory divided by loss rate) are substantially shorter (1–3 years) than the age of standing fine root C stocks obtained from radiocarbon (4–11 years). By assuming positively skewed distributions for root ages, we can effectively decouple the mean age of C in live fine roots (measured using ¹⁴C) from the rate of C flow through the live root pool, and resolve these apparently disparate estimates of root C dynamics. Explaining the ¹⁴C values in soil pore space CO₂, in addition, requires that a portion of the decomposing roots be cycled through soil organic matter pools with decadal turnover time.     

Isolation, Characterization, and Transcriptional Analysis of the Chitinase chi2 Gene (DQ011663) from the Biocontrol Fungus Metarhizium anisopliae var. anisopliae

Metarhizium anisopliae infects arthropods via a combination of specialized structures and cuticle degradation. Hydrolytic enzymes are accepted as key factors for the host penetration step and include chitinases. The characterization of the chi2 chitinase gene from M. anisopliae var. anisopliae is reported. The chi2 gene is interrupted by two short introns and is 1,542-bp long, coding a predicted protein of 419 amino acids with a stretch of 19 amino acid residues displaying characteristics of signal peptide. The predicted chitinase molecular mass is 44 kDa with a mature protein of 42 kDa and a theoretical pI of 4.8. The comparison of the CHI2 predicted protein to fungal orthologues revealed similarity to the glycohydrolase family 18 and a phylogenetic analysis was conducted. The chi2 gene is up-regulated by chitin as a carbon source and in conditions of fungus autolysis, and is down-regulated by glucose. This regulation is consistent with the presence of putative CreA/Crel/Crr1 carbon catabolic repressor binding domains on the regulatory sequence.     

The effect of Bulgarian propolis against Trypanosoma cruzi and during its interaction with host cells

Propolis has shown activity against pathogenic microorganisms that cause diseases in humans and animals. The ethanol (Et-Blg) and acetone (Ket-Blg) extracts from a Bulgarian propolis, with known chemical compositions, presented similar activity against tissue culture-derived amastigotes. The treatment of Trypanosoma cruzi-infected skeletal muscle cells with Et-Blg led to a decrease of infection and of the intracellular proliferation of amastigotes, while damage to the host cell was observed only at concentration 12.5 times higher than those affecting the parasite. Ultrastructural analysis of the effect of both extracts in epimastigotes revealed that the main targets were the mitochondrion and reservosomes. Et-Blg also affected the mitochondrion-kinetoplast complex in trypomastigotes, offering a potential target for chemotherapeutic agents.     

Cell proliferation and interferon-gamma response to recombinant MBP-3, NarL, MT-10.3, and 16 kDa Mycobacterium tuberculosis antigens in Brazilian tuberculosis patients

Human pulmonary tuberculosis (TB) is a worldwide public health problem. In resistant individuals, control of the infection mainly requires development of a Th1 cell immune response with production of cytokines, of which interferon-gamma (IFN-gamma)plays an important role. Several antigens from Mycobacterium tuberculosis complex has been described for use in vaccine development or for diagnostic purposes, however little evaluation has been done in endemic area for TB. The proliferative and IFN-gamma human T cell immune responses, to four recombinant proteins (MBP-3, NarL, MT-10.3, 16 kDa) and PPD, of 38 Brazilian TB patients (6 untreated and 32 treated) and 67 controls (38 positive and 29 negative tuberculin skin test - TST) were compared. The highest reactivity mean rate was obtained with PPD followed by 16 kDa in TB patients. While most of the patients (87%) and controls (> 64%) respond to the PPD, 16 kDa was more specifically recognized (> 21%) although less sensitive (54%). When TB patients were divided according to treatment status, opposite to PPD, higher average level of IFN-gamma was induced by 16 kDa in untreated (505 pg/ml) compared to treated TB patients and TST+ (269.8 pg/ml x 221.6 pg/ml, respectively), although the difference was not significant. These data show that in contrast with the other recombinant proteins, the stimulatory potency of 16 kDa to induce proliferative and INF-gamma response was more effective and is more recognized by active TB untreated patients, eliciting in control individuals a more selective immune response than PPD.     

The acidic domain of hnRNPQ (NSAP1) has structural similarity to Barstar and binds to Apobec1

Apobec1 edits the ApoB mRNA by deaminating nucleotide C(6666), which results in a codon change from Glutamate to stop, and subsequent expression of a truncated protein. Apobec1 is regulated by ACF (Apobec1 complementation factor) and hnRNPQ, which contains an N-terminal "acidic domain" (AcD) of unknown function, three RNA recognition motifs, and an Arg/Gly-rich region. Here, we modeled the structure of AcD using the bacterial protein Barstar as a template. Furthermore, we demonstrated by in vitro pull-down assays that 6xHis-AcD alone is able to interact with GST-Apobec1. Finally, we performed in silico phosphorylation of AcD and molecular dynamics studies, which indicate conformational changes in the phosphorylated form. The results of the latter studies were confirmed by in vitro phosphorylation of 6xHis-AcD by protein kinase C, mass spectrometry, and spectroscopic analyses. Our data suggest hnRNPQ interactions via its AcD with Apobec1 and that this interaction is regulated by the AcD phosphorylation.     

eNOS genotype-dependent correlation between whole blood lead and plasma nitric oxide products concentrations.

Experimental data indicate that lead exposure decreases nitric oxide (NO) availability. However, no previous study has examined whether lead exposure affects plasma nitrite/nitrate (NO(x)) concentrations in humans. In addition, the T(-786)C polymorphism affects endothelial NO synthase (eNOS) expression and endogenous NO release. Here, we investigated whether there is an association between the circulating concentrations of NO(x) and the concentrations of lead in whole blood (B-Pb) and in plasma (P-Pb) from lead-exposed subjects. In addition, we also evaluated whether eNOS genotype for the T(-786)C polymorphism affects NO(x) concentrations in lead-exposed subjects. We studied 104 subjects exposed to lead who were non-smokers, 18-60 years of age, and not alcohol consumers. Genomic DNA was isolated from blood samples and genotypes for the T(-786)C polymorphism were determined by PCR and restriction fragment length digestion. Circulating NO(x) was determined by chemiluminescence. B-Pb and P-Pb were determined by inductively coupled plasma mass spectrometry and by graphite furnace atomic absorption spectrometry, respectively. No significant correlations were found between NO(x) and B-Pb and P-Pb measured in the 104 subjects (all P > 0.05). However, while no significant correlation was found in subjects with TT genotype, a negative correlation was found between plasma NO(x) and B-Pb (r = 0.230, P = 0.048) and P-Pb (r = 0.194, P = 0.110) in subjects from TC + CC genotypes group. Our study shows a negative correlation between plasma NO(x) concentrations and B-Pb in carriers of the "C" allele, thus suggesting a possible mechanism possibly involved in lead exposure-induced increase in the susceptibility to cardiovascular diseases.     

Expression systems for heterologous production of antimicrobial peptides

Antimicrobial peptides (AMPs) consist of molecules that act on the defense systems of numerous organisms toward multiple pathogens such as bacteria, fungi, parasites and viruses. These compounds have become extremely significant due to the increasing resistance of microorganisms to common antibiotics. However, the low quantity of peptides obtained from direct purification is, to date, still a remarkable bottleneck for scientific and industrial research development. Therefore, this review describes the main heterologous systems currently used for AMP production, including bacteria, fungi and plants, and also the related strategies for reaching greater functional peptide production. The main difficulties of each system are also described in order to provide some directions for AMP production. In summary, data revised here indicate that large-scale production of AMPs can be obtained using biotechnological tools, and the products may be applied in the pharmaceutical industry as well as in agribusiness.     

Effects of direct electric current on Herpetomonas samuelpessoai: An ultrastructural study

The literature shows that the effects of direct electric currents on biological material are numerous, including bactericidal, fungicidal, parasiticidal, and anti‐tumoral, among others. Non‐pathogenic trypanosomatids, such as Herpetomonas samuelpessoai, have emerged as important models for the study of basic biological processes performed by a eukaryotic cell. The present study reports a dose‐dependent anti‐protozoan effect of direct electric treatment with both cathodic and anodic current flows on H. samuelpessoai cells. The damaging effects can be attributable to the electrolysis products generated during electric stimulation. The pH of the cell suspension was progressively augmented from 7.4 to 10.5 after the cathodic treatment. In contrast, the anodic treatment caused a pH decrease varying from 7.4 to 6.5. Transmission electron microscopy analyses revealed profound alterations in vital cellular structures (e.g., mitochondrion, kinetoplast, flagellum, flagellar pocket, nucleus, and plasma membrane) after exposure to both cathodic and anodic current flows. Specifically, cathodic current flow treatment induced the appearance of autophagic‐like structures on parasite cells, while those submitted to an anodic current flow presented marked disorganization of plasma membrane and necrotic appearance. However, parasites treated in the intermediary chamber (without contact with the electrodes) did not present significant changes in viability or morphology, and no pH variation was detected in this system. The use of H. samuelpessoai as a biological model and the direct electric current experimental approach used in our study provide important information for understanding the mechanisms involved in the cytotoxic effects of this physical agent. Bioelectromagnetics 33:334–345, 2012. © 2011 Wiley Periodicals, Inc.