Evaluation of the catalytic specificity,biochemical properties,and milk clotting abilities of an aspartic peptidase from <Emphasis Type="Italic">Rhizomucor miehei</Emphasis>

In this study, we detail the specificity of an aspartic peptidase from Rhizomucor miehei and evaluate the effects of this peptidase on clotting milk using the peptide sequence of k-casein (Abz-LSFMAIQ-EDDnp) and milk powder. Molecular mass of the peptidase was estimated at 37 kDa, and optimum activity was achieved at pH 5.5 and 55 °C. The peptidase was stable at pH values ranging from 3 to 5 and temperatures of up 45 °C for 60 min. Dramatic reductions in proteolytic activity were observed with exposure to sodium dodecyl sulfate, and aluminum and copper (II) chloride. Peptidase was inhibited by pepstatin A, and mass spectrometry analysis identified four peptide fragments (TWSISYGDGSSASGILAK, ASNGGGGEYIFGGYDSTK, GSLTTVPIDNSR, and GWWGITVDRA), similar to rhizopuspepsin. The analysis of catalytic specificity showed that the coagulant activity of the peptidase was higher than the proteolytic activity and that there was a preference for aromatic, basic, and nonpolar amino acids, particularly methionine, with specific cleavage of the peptide bond between phenylalanine and methionine. Thus, this peptidase may function as an important alternative enzyme in milk clotting during the preparation of cheese.     

Recent advances in analytical procedures for the detection of diarrhetic phycotoxins: a review

Okadaic acid and dinophysistoxins are produced by some marine unicellular algae from the plankton and also benthic microalgae and may accumulate in shellfish. These phycotoxins are involved in a gastrointestinal syndrome called diarrhetic shellfish poisoning (DSP), which occurs in humans after consumption of bivalve molluscs. Thousands cases of human poisonings in Europe were caused by consumption of toxic shellfish during the past decade. The rapid detection and the reliable determination of the main phycotoxins implicated in DSP are a major concern for governmental institutions in charge of the sanitary control of seafood safety. Analytical procedures for the detection and determination of DSP toxins can be classified as: bioassays, biochemical methods including immunoassays, or physicochemical methods. Although a large number of methods have been developed, none have been officially validated. A complete panel of tools for DSP toxin analysis should include screening, investigation, and confirmation methods. This paper presents a compilation of recent developments and optimisations of these methods. This revised version was published online in June 2006 with corrections to the Cover Date.     

Immobilization and health assessment of free-ranging black spider monkeys (Ateles paniscus chamek)

Eight free-ranging black spider monkeys (Ateles paniscus chamek) were immobilized with Telazol^® in Bolivia for the purpose of radio-collaring. During this procedure, the animals received complete medical examinations, and samples were collected for health analyses. Biochemical test results varied with the degree of condition of the animals, and a variety of physical abnormalities were found. Evidence of previous infections with Leptospira sp., encephalitis virus, and yellow fever virus was found. All findings contribute to establishing baseline health values for the species. The handling of primates for research projects provides a valuable opportunity to collect health-related data and samples that can contribute to wildlife management and conservation efforts. The capture and handling of free-ranging primates is always accompanied by risk of injury or mortality. It is ethically important to maximize the amount of information gathered during these procedures. Furthermore, sharing the undesirable impacts with the scientific community enables informed decisions to be made during future project development. Am. J. Primatol. 44:107–123, 1998. © 1998 Wiley-Liss, Inc.     

Primate diversity,habitat preferences,and population density estimates in Noel Kempff Mercado National Park,Santa Cruz Department,Bolivia

This report documents primate communities at two sites within Noel Kempff Mercado National Park in northeastern Santa Cruz Department, Bolivia. Diurnal line transects and incidental observations were employed to survey two field sites, Lago Caiman and Las Gamas, providing information on primate diversity, habitat preferences, relative abundance, and population density. Primate diversity at both sites was not particularly high, with six observed species: Callithrix argentata melanura, Aotus azarae, Cebus apella, Alouatta caraya, A. Seniculus, and Ateles paniscus chamek. Cebus showed no significant habitat preferences at Lago Caiman and was also more generalist in use of forest strata, whereas Ateles clearly preferred the upper levels of structurally tall forest. Callithrix argentata melanura was rarely encountered during surveys at Lago Caiman, where it preferred low vine forest. Both species of Alouatta showed restricted habitat use and were sympatric in Igapo forest in the Lago Caiman area. The most abundant primate at both field sites was Ateles, with density estimates reaching 32.1 individuals/km² in the lowland forest at Lago Caiman, compared to 14.1 individuals/km² for Cebus. Both Ateles and Cebus were absent from smaller patches of gallery forest at Las Gamas. These densities are compared with estimates from other Neotropical sites. The diversity of habitats and their different floristic composition may account for the numerical dominance of Ateles within the primate communities at both sites. Am. J. Primatol. 46:197–211, 1998. © 1998 Wiley-Liss, Inc.     

microRNA159‐targeted SlGAMYB transcription factors are required for fruit set in tomato

The transition from flowering to fruit production, namely fruit set, is crucial to ensure successful sexual plant reproduction. Although studies have described the importance of hormones (i.e. auxin and gibberellins) in controlling fruit set after pollination and fertilization, the role of microRNA‐based regulation during ovary development and fruit set is still poorly understood. Here we show that the microRNA159/GAMYB1 and ‐2 pathway (the miR159/GAMYB1/2 module) is crucial for tomato ovule development and fruit set. MiR159 and SlGAMYBs were expressed in preanthesis ovaries, mainly in meristematic tissues, including developing ovules. SlMIR159‐overexpressing tomato cv. Micro‐Tom plants exhibited precocious fruit initiation and obligatory parthenocarpy, without modifying fruit shape. Histological analysis showed abnormal ovule development in such plants, which led to the formation of seedless fruits. SlGAMYB1/2 silencing in SlMIR159‐overexpressing plants resulted in misregulation of pathways associated with ovule and female gametophyte development and auxin signalling, including AINTEGUMENTA‐like genes and the miR167/SlARF8a module. Similarly to SlMIR159‐overexpressing plants, SlGAMYB1 was downregulated in ovaries of parthenocarpic mutants with altered responses to gibberellins and auxin. SlGAMYBs likely contribute to fruit initiation by modulating auxin and gibberellin responses, rather than their levels, during ovule and ovary development. Altogether, our results unveil a novel function for the miR159‐targeted SlGAMYBs in regulating an agronomically important trait, namely fruit set.     

Predicting binding modes of reversible peptide‐based inhibitors of falcipain‐2 consistent with structure–activity relationships

Falcipain‐2 (FP‐2) is a major hemoglobinase of Plasmodium falciparum, considered an important drug target for the development of antimalarials. A previous study reported a novel series of 20 reversible peptide‐based inhibitors of FP‐2. However, the lack of tridimensional structures of the complexes hinders further optimization strategies to enhance the inhibitory activity of the compounds. Here we report the prediction of the binding modes of the aforementioned inhibitors to FP‐2. A computational approach combining previous knowledge on the determinants of binding to the enzyme, docking, and postdocking refinement steps, is employed. The latter steps comprise molecular dynamics simulations and free energy calculations. Remarkably, this approach leads to the identification of near‐native ligand conformations when applied to a validation set of protein‐ligand structures. Overall, we proposed substrate‐like binding modes of the studied compounds fulfilling the structural requirements for FP‐2 binding and yielding free energy values that correlated well with the experimental data. Proteins 2017; 85:1666–1683. © 2017 Wiley Periodicals, Inc.     

Shared or distinct responses between intermediate and satellite stream fish species in an altered Amazonian River?

Environmental and spatial variables can distinctly influence the occupancy frequency distributions in stream fish. From a metacommunity context, we tested the following hypothesis, intermediate species are governed by dispersal and niche-based processes; in contrast, satellite species are governed by niche-based processes. To test this, we separately analyzed three data sets, the entire metacommunity, the intermediate species and the satellite species, using a forward selection of explanatory variables, and a partial Redundancy Distance Analysis. The fish and 31 variables of 52 stream reaches of a Brazilian river basin in the Western Amazon were collected during the dry period of 2012. The results for all of the data set revealed two different patterns: on one side, satellite species revealed that niche and dispersal-based processes were the most important; on the other side, for intermediate species and for all of the species set, only dispersal-based processes were the most important. For the data set including all of the species and the intermediate species, the variance was explained mainly by landscape scale variables. By contrast, the variance within the satellite species set was explained by local scale variables. Management efforts for intermediate species should be taking at larger scale, but they are usually less critical for the maintenance of aquatic biodiversity; on the other hand, management efforts for satellite species should be taken at smaller scale and based on specific biological and ecological information for the focal species.     

Extension of coronary artery disease is associated with increased IL-6 and decreased adiponectin gene expression in epicardial adipose tissue

Epicardial adipose tissue (EAT) expresses lower levels of adiponectin in patients with CAD and higher levels of inflammatory mediators such as IL-6 and leptin than subcutaneous adipose tissue. This showed one important role of EAT in coronary artery disease. However, the relationship of EAT adiponectin and IL-6 levels to the extension of coronary artery disease has not hitherto been determined. We sought to determine whether the levels of adiponectin and interleukin-6 (IL-6) mRNA in epicardial adipose tissue are associated with the extension of coronary artery disease (CAD). Methods: Angiographic and hormones expression were evaluated from epicardial and subcutaneous adipose tissue. 92 patients (58 CAD, 34 non-CAD) who underwent cardiac surgery. Adiponectin and IL-6 mRNA levels were measured by real time RT-PCR in epicardial and subcutaneous adipose tissue (SAT) following angiographic evaluation of their coronary arteries. Results: We found that epicardial adipose tissue of CAD expressed lower levels of adiponectin mRNA and higher levels of IL-6 mRNA than that of non-CAD patients. As the number of injured arteries rose, adiponectin mRNA levels decreased (r = −0.402, p < 0.001) and IL-6 mRNA increased (r = 0.514, p < 0.001) in epicardial adipose tissue. Conclusions: The extension of CAD is significantly associated with the expression of adiponectin and IL-6 mRNA in EAT. These findings suggest that low adiponectin and high IL-6 expression by EAT may contribute to CAD extension.