Biochemical characterization of digestive membrane‐associated alkaline phosphatase from the velvet bean caterpillar Anticarsia gemmatalis

In Brazil, the use of transgenic plants expressing the insect‐toxic Bacillus thuringiensis endotoxin has been successfully used as pest control management since 2013 in transgenic soybean lineages against pest caterpillars such as Helicoverpa armigera. These toxins, endogenously expressed by the plants or sprayed over the crops, are ingested by the insect and bind to receptors in the midgut of these animals, resulting in disruption of digestion and lower insect survival rates. Here, we identified and characterized a membrane‐associated alkaline phosphatase (ALP) in the midgut of Anticarsia gemmatalis, the main soybean defoliator pest in Brazil, and data suggested that it binds to Cry1Ac toxin in vitro. Our data showed a peak of ALP activity in homogenate samples of the midgut dissected from the 4th and 5th instars larvae. The brush border membrane vesicles obtained from the midgut of these larvae were used to purify a 60 kDa ALP, as detected by in‐gel activity and in vitro biochemical characterization using pharmacological inhibitors and mass spectrometry. When Cry1Ac toxin was supplied to the diet, it was efficient in decreasing larval weight gain and survival. Indeed, in vitro incubation of Cry1Ac toxin with the purified ALP resulted in a 43% decrease in ALP specific activity and enzyme‐linked immunosorbent assay showed that ALP interacts with Cry1Ac toxin in vitro, thus suggesting that ALP could function as a Cry toxin ligand. This is a first report characterizing an ALP in A. gemmatalis.     

Towards an absolute scale for adhesion strength of ship hull microfouling

In-water ships’ hull cleaning enables significant fuel savings through removal of marine fouling from surfaces. However, cleaning may also shorten the lifetime of hull coatings, with a subsequent increase in the colonization and growth rate of fouling organisms. Deleterious effects of cleaning would be minimized by matching cleaning forces to the adhesion strength of the early stages of fouling, or microfouling. Calibrated waterjets are routinely used to compare different coatings in terms of the adhesion strength of microfouling. However, an absolute scale is lacking for translating such results into cleaning forces, which are of interest for the design and operation of hull cleaning devices. This paper discusses how such forces can be determined using computational fluid dynamics. Semi-empirical formulae are derived for forces under immersed waterjets, where the normal and tangential components of wall forces are given as functions of different flow parameters. Nozzle translation speed is identified as a parameter for future research, as this may affect cleaning efficacy.     

Synthesis and biological evaluation of a small library of nojirimycin-derived bicyclic iminosugars

Novel nojirimycin-derived bicyclic structures, containing cyclic carbamate, urea and guanidine moieties have been synthesised starting from suitably protected alpha-C-vinylnojirimycin and alpha-C-allylnojirimycin, respectively, and their biological activity against different glycosidases and as antibacterial agents tested.     

Assessing the effects of surfactants on the physical properties of liposome membranes

Knowledge of the partition process of environmentally significant molecules between biological membranes and their surroundings is of vital importance to explain their activity and toxicity, as well as phenomena like absorption, distribution and metabolism. In this research effort, we have studied membrane interactions of three surfactants: t-octylphenoxypolyethoxyethanol (Triton X-100), cetyltrimethylammonium chloride (CTAC) and dodecylbenzene sulphonate (SDBS). Unilamellar liposomes (LUVs) of egg yolk phosphatidylcholine (EPC) were used as membrane models. The partition coefficient, a fundamental parameter in assessing the behaviour of xenobiotic compounds, was determined for SDBS and Triton X-100 by derivative spectrophotometry and fluorescence quenching. The effect of these surfactants upon the physico-chemical characteristics (fluidity, diameter and surface charge) of the liposome membrane was also determined. Results show that all the three surfactants cause an increase in fluidity of the liposome membrane, although for low surfactant concentrations uncharacteristic membrane rigidity was observed, probably due to a change in lipid packing density.     

Leftward Shift in the Voltage-Dependence for Ca2+ Currents Activation Induced by a New Toxin from Phoneutria reidyi (Aranae, Ctenidae) Venom

Summary Various neurotoxins have been described from the venom of the Brazilian spider Phoneutria nigriventer, but little is known about the venoms of the other species of this genus. In the present work, we describe the purification and some structural and pharmacological features of a new toxin (PRTx3-7) from Phoneutria reidyi that causes flaccid paralysis in mice. The observed molecular mass (4627.26 Da) was in accordance with the calculated mass for the amidated form of the amino acid sequence (4627.08 Da). The presence of an α-amidated C-terminus was confirmed by MS/MS analysis of the C-terminal peptide, isolated after enzymatic digestion of the native protein with Glu-C endoproteinase. The purified protein was injected (intracerebro-ventricular) into mice at dose levels of 5 μg/mouse causing immediate agitation and clockwise gyration, followed by the gradual development of general flaccid paralysis. PRTx3-7 at 1 μM inhibited by 20% the KCl-induced increase on [Ca²⁺]_i in rat brain synaptosomes. The HEK cells permanently expressing L, N, P/Q and R HVA Ca²⁺ channels were also used to better characterize the pharmacological features of PRTx3-7. To our surprise, PRTx3-7 shifted the voltage-dependence for activation towards hyperpolarized membrane potentials for L (−4 mV), P/Q (−8 mV) and R (−5 mV) type Ca²⁺ currents. In addition, the new toxin also affected the steady state of inactivation of L-, N- and P/Q-type Ca²⁺ currents. L. B. Vieira and A. M. C. Pimenta contributed equally to this work.     

An improved method for two-dimensional fluorescence monitoring of complex bioreactors

An improved method for deconvoluting complex spectral maps from bidimensional fluorescence monitoring is presented, relying on a combination of principal component analysis (PCA) and feedforward artificial neural networks (ANN). With the aim of reducing ANN complexity, spectral maps are first subjected to PCA, and the scores of the retained principal components are subsequently used as ANN input vector. The method is presented using the case study of an extractive membrane biofilm reactor, where fluorescence maps of a membrane-attached biofilm were analysed, which were collected under different reactor operating conditions. During ANN training, the spectral information is associated with process performance indicators. Originally, 231 excitation/emission pairs per fluorescence map were used as ANN input vector. Using PCA, each fluorescence map could be represented by a maximum of six principal components, thereby catching 99.5% of its variance. As a result, the dimension of the ANN input vector and hence the complexity of the artificial neural network was significantly reduced, and ANN training speed was increased. Correlations between principal components and ANN predicted process performance parameters were good with correlation coefficients in the order of 0.7 or higher.     

Divergence or disorder?: the politics of naming intersex

The conditions once known under the umbrella terms intersex and hermaphroditism are now generally being called disorders of sex development in medical settings. The terms might seem synonymous, but in fact there are significant differences with controversial consequences. Hermaphroditism, an older term that can still be found in many medical writings, is vague, demeaning, and sensationalistic, conjuring mythic images of monsters and freaks. In the 1990s, activists advocated intersex to describe discordance between the multiple components of sex anatomy, but that word alienated many parents of affected children, as it suggests a self-conscious alternative gender identity and sexuality. Disorders of sex development also refers to intersex, but it deemphasizes the identity politics and sexual connotations associated with intersex, avoids the degradation associated with hermaphrodite, and instead highlights the underlying genetic or endocrine factors that cause prenatal sex development to take an unusual path. I argue that using disorder is problematic, because it implies medical conditions in need of repair, when some intersex anatomies, though atypical, do not necessarily need surgical or hormonal correction. I advocate a less pathologizing new term, divergence of sex development, that might reduce some of the conflict over nomenclature and satisfy intersex people, their parents, and their doctors.     

Talaromyces amestolkiae uses organic phosphate sources for the treatment of uranium-contaminated water

BioMetals - Fungi have received particular attention in regards to alternatives for bioremediation of heavy metal contaminated locales. Enzymes produced by filamentous fungi, such as phosphatases,...