Oxygen stable isotope ratios of tree-ring cellulose: the next phase of understanding

Analysis of the oxygen isotope ratio of tree-ring cellulose is a valuable tool that can be used as a paleoclimate proxy. Our ability to use this tool has gone through different phases. The first began in the 1970s with the demonstration of empirical relationships between the oxygen isotope ratio of tree-ring cellulose and climate. These empirical relationships, however, did not provide us with the confidence that they are robust through time, across taxa and across geographical locations. The second phase began with a rudimentary understanding of the physiological and biochemical mechanisms responsible for the oxygen isotope ratios of cellulose, which is necessary to increase the power of this tool. This phase culminated in a mechanistic tree-ring model integrating concepts of physiology and biochemistry in a whole-plant system. This model made several assumptions about leaf water isotopic enrichment and biochemistry which, in the nascent third phase, are now being challenged, with surprising results. These third-phase results suggest that, contrary to the model assumption, leaf temperature across a large latitudinal gradient is remarkably constant and does not follow ambient temperature. Recent findings also indicate that the biochemistry responsible for the incorporation of the cellulose oxygen isotopic signature is not as simple as has been assumed. Interestingly, the results of these challenges have strengthened the tree-ring model. There are several other assumptions that can be investigated which will improve the utility of the tree-ring model.     

A multimetric index based on benthic macroinvertebrates for evaluation of Atlantic Forest streams at Rio de Janeiro State, Brazil

This study describes the application of a protocol for biological assessment of water quality at first to third order streams at Serra dos órg?os, an area covered by Atlantic Forest in Rio de Janeiro State, Brazil. Major impacts in the region are domestic effluents and deforestation. Our main objective is to establish biocriteria for the establishment of the Serra dos órg?os Multimetric Index (SOMI) based on benthic macroinvertebrates. We used data from previous studies, sampled by experienced biologists, from 1999 through 2002. The benthic macroinvertebrate community was sampled in 12 reference sites and seven impaired sites in three river basins: Guapimirim, Macaé and Grande, all from the same bioregion. From the 22 tested metrics, 6 were included in the SOMI (% Diptera, % Coleoptera, Family Taxa, EPT Taxa, BMWP-CETEC and % Shredders). Scores (5, 3 or 1) were developed for these metrics to allow for aggregation into the index. Seven intermediately impaired sites were used for evaluating the applicability of the multimetric index. We concluded that the SOMI is a robust easy-to-apply tool for biomonitoring programs in the Serra dos órg?os region, south-east Brazil. Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorised users. Handling editor: D. Dudgeon     

A phylogenetic study of the Thygater‐Trichocerapis group and new scopes for the subgenera of Thygater Holmberg, (Hymenoptera,Apidae)

The bee tribe Eucerini is a large monophyletic taxon occurring in all continents, except Oceania and Antarctica, but far more diverse in the Americas and, especially, in the Neotropics. The phylogenetic relationships within its subtribe Eucerina, which contains the bulk of Eucerini diversity, is poorly understood, this being especially true for the relationships among its Neotropical representatives, which have been poorly represented in all phylogenetic studies, including Eucerini, to date. This leads to a generalized lack of confidence on the monophyly of the groups currently accepted as genera and subgenera. Here, a phylogenetic study based on three molecular markers (COI, 18S, and 28S, totalling about 1700 bp) and 58 morphological characters is presented as a contribution to the understanding of the relationships of the so‐called Thygater‐Trichocerapis group, and especially of the genus Thygater Holmberg, which has not previously been extensively sampled. Representatives of Trichocerapis, including its monotypic subgenus T. (Dithygater), are included for the first time in a phylogenetic study. The main results were: (i) support for the monophyly of the Thygater‐Trichocerapis group; (ii) support for the monophyly of Thygater; (iii) recognition of two main clades in Thygater, each one containing the type species of one of the previously recognized subgenera; and (iv) additional support for the position of Alloscirtetica as sister to all remaining Eucerina. Based on these results a redefinition of the scope of the two subgenera of Thygater is proposed, with changes in the subordination of three of its species, T. (Nectarodiaeta) chaetaspis comb.n. , T. (Nectarodiaeta) paranaensis comb.n. and T. (Thygater) mexicana comb.n.     

Mucosal layers and related nerve fibres in non-chagasic and chagasic human colon—a quantitative immunohistochemical study

Chagasic megacolon is accompanied by extensive myenteric and, simultaneously, moderate submucosal neuron loss. Here, we examined changes of the innervation pattern of the lamina propria (LP) and muscularis mucosae (MM). Two alternating sets of cryosections were taken from seven non-chagasic colonic and seven chagasic megacolonic specimens (the latter included both the dilated megacolonic and the non-dilated transitional oral and anal zones) and were immunohistochemically triple-stained for smooth-muscle actin (SMA), synaptophysin (SYN) and glial acid protein S100 and, alternatively, for SMA, vasoactive intestinal peptide (VIP) and somatostatin (SOM). Subsequent image analysis and statistical evaluation of nervous tissue profile areas revealed that, in LP, the most extreme differences (i.e. increase in thickness or decrease in nerve, glia and muscle tissue profile area, respectively) compared with control values occurred in the dilated megacolonic zone itself. In contrast, the most extreme differences in the MM were in the anal-to-megacolonic zone (except the profile area of muscle tissue, which was lowest in the megacolonic zone). This parallels our previous results in the external muscle coat. A partial and selective survival of VIP-immunoreactive in contrast to SOM-immunoreactive nerve fibres was observed in both mucosal layers investigated. Thus, VIPergic nerve elements might be crucial for the maintenance of the mucosal barrier. The differential changes of neural tissue parameters in LP and MM might reflect a multifactorial rather than a pure neurogenic development of megacolon in chronic Chagas’ disease.     

Bromelain purification through unconventional aqueous two-phase system (PEG/ammonium sulphate)

This paper focuses on the feasibility of unconventional aqueous two-phase systems for bromelain purification from pineapple processing waste. The main difference in comparison with conventional systems is the integration of the liquid–liquid extraction technique with fractional precipitation, which can decrease the protein content with no loss of biological activity by removing of unwanted molecules. The analysis of the results was based on the response surface methodology and revealed that the use of the desirability optimisation methodology (DOM) was necessary to achieve higher purification factor values and greater bromelain recovery. The use of DOM yielded an 11.80-fold purification factor and 66.38 % biological activity recovery using poly(ethylene glycol) (PEG) with a molar mass of 4,000, 10.86 % PEG concentration (m/m) and 36.21 % saturation of ammonium sulphate.     

Visual Evoked Cortical Potential (VECP) Elicited by Sinusoidal Gratings Controlled by Pseudo-Random Stimulation

The contributions of contrast detection mechanisms to the visual cortical evoked potential (VECP) have been investigated studying the contrast-response and spatial frequency-response functions. Previously, the use of m-sequences for stimulus control has been almost restricted to multifocal electrophysiology stimulation and, in some aspects, it substantially differs from conventional VECPs. Single stimulation with spatial contrast temporally controlled by m-sequences has not been extensively tested or compared to multifocal techniques. Our purpose was to evaluate the influence of spatial frequency and contrast of sinusoidal gratings on the VECP elicited by pseudo-random stimulation. Nine normal subjects were stimulated by achromatic sinusoidal gratings driven by pseudo random binary m-sequence at seven spatial frequencies (0.4–10 cpd) and three stimulus sizes (4°, 8°, and 16° of visual angle). At 8° subtence, six contrast levels were used (3.12–99%). The first order kernel (K1) did not provide a consistent measurable signal across spatial frequencies and contrasts that were tested–signal was very small or absent–while the second order kernel first (K2.1) and second (K2.2) slices exhibited reliable responses for the stimulus range. The main differences between results obtained with the K2.1 and K2.2 were in the contrast gain as measured in the amplitude versus contrast and amplitude versus spatial frequency functions. The results indicated that K2.1 was dominated by M-pathway, but for some stimulus condition some P-pathway contribution could be found, while the second slice reflected the P-pathway contribution. The present work extended previous findings of the visual pathways contribution to VECP elicited by pseudorandom stimulation for a wider range of spatial frequencies.     

Compound A398, a Novel Podophyllotoxin Analogue: Cytotoxicity and Induction of Apoptosis in Human Leukemia Cells

Despite advances in oncology research, cancer is one of the leading causes of death worldwide. Thus, there is a demand for the development of more selective and effective antitumor agents. This study showed that A398, a novel podophyllotoxin analogue, was cytotoxic to the HT-29, MCF-7, MOLT-4 and HL-60 tumor cell lines, being less active in human peripheral blood mononuclear cells and normal cell lines FGH and IEC-6. Tests using the HepG2 lineage indicated that its metabolites do not contribute to its cytotoxicity. In the HL-60 cells, A398 induced apoptosis in a time and concentration-dependent manner, promoting mitochondrial depolarization, inhibition of Bcl-2, phosphatidylserine exposure, activation of caspases -8, -9 and -3, and DNA fragmentation. The production of reactive oxygen species does not seem to be a crucial event for the apoptotic process. Pretreatment with specific inhibitors of kinases ERK1/2, JNK and p38 resulted in an increased percentage of death induced by A398. These results indicate that the compound induced apoptosis through activation of intrinsic and extrinsic death pathways with the mechanism involving the inhibition of the MAPKs and Bcl-2. Taken together, our findings suggest that A398 has an anticancer potential, proving itself to be a candidate for preclinical studies.