High-Throughput Screening of a Marine Compound Library Identifies Anti-Cryptosporidium Activity of Leiodolide A

Cryptosporidium sp. are apicomplexan parasites that cause significant morbidity and possible mortality in humans and valuable livestock. There are no drugs on the market that are effective in the population most severely affected by this parasite. This study is the first high-throughput screen for potent anti-Cryptosporidium natural products sourced from a unique marine compound library. The Harbor Branch Oceanographic Institute at Florida Atlantic University has a collection of diverse marine organisms some of which have been subjected to medium pressure liquid chromatography to create an enriched fraction library. Numerous active compounds have been discovered from this library, but it has not been tested against Cryptosporidium parvum. A high-throughput in vitro growth inhibition assay was used to test 3764 fractions in the library, leading to the identification of 23 fractions that potently inhibited the growth of Cryptosporidium parvum. Bioassay guided fractionation of active fractions from a deep-sea sponge, Leiodermatium sp., resulted in the purification of leiodolide A, the major active compound in the organism. Leiodolide A displayed specific anti-Cryptosporidium activity at a half maximal effective concentration of 103.5 nM with selectivity indexes (SI) of 45.1, 11.9, 19.6 and 14.3 for human ileocecal colorectal adenocarcinoma cells (HCT-8), human hepatocellular carcinoma cells (Hep G2), human neuroblastoma cells (SH-SY5Y) and green monkey kidney cells (Vero), respectively. The unique structure of leiodolide A provides a valuable drug scaffold on which to develop new anti-Cryptosporidium compounds and supports the importance of screening natural product libraries for new chemical scaffolds.     

Effect of Electric Field Treatment on Unsaturated Fatty Acid in Crude Avocado Oil

The objective of this study was to evaluate the stability of the fatty acids in avocado oil when the product is subjected to different conditions of electric field treatment (voltage: 5 kV cm^?1; frequency: 720 Hz; treatment time: 5, 10, 15, 20, and 25 min). Fatty acids were analyzed by Fourier transform infrared spectroscopy in the mid-infrared region. Electric field is a suitable method to preserve the oil quality and composition with minimal modifications in unsaturated fatty acids.     

Activation of the Tumor Suppressor PP2A Emerges as a Potential Therapeutic Strategy for Treating Prostate Cancer

Protein phosphatase 2A (PP2A) is a tumor suppressor complex that has recently been reported as a novel and highly relevant molecular target in prostate cancer (PCa). However, its potential therapeutic value remains to be fully clarified. We treated PC-3 and LNCaP cell lines with the PP2A activators forskolin and FTY720 alone or combined with the PP2A inhibitor okadaic acid. We examined PP2A activity, cell growth, prostasphere formation, levels of PP2A phosphorylation, CIP2A and SET expression, and AKT and ERK activation. Interestingly, both forskolin and FTY720 dephosphorylated and activated PP2A, impairing proliferation and prostasphere formation and inducing changes in AKT and ERK phosphorylation. Moreover, FTY720 led to reduced CIP2A levels. Treatment with okadaic acid impaired PP2A activation thus demonstrating the antitumoral PP2A-dependent mechanism of action of both forskolin and FTY720. Levels of PP2A phosphorylation together with SET and CIP2A protein expression were studied in 24 PCa patients and both were associated with high Gleason scores and presence of metastatic disease. Altogether, our results suggest that PP2A inhibition could be involved in PCa progression, and the use of PP2A-activating drugs might represent a novel alternative therapeutic strategy for treating PCa patients.     

The Marine Natural Product Furospinulosin 1 Induces Apoptosis in MDA-MB-231 Triple Negative Breast Cancer Cell Spheroids,But Not in Cells Grown Traditionally with Longer Treatment

Cancer cells grown in spheroid conditions interact with each other and the extracellular matrix, providing a better representation of the in vivo environment than two-dimensional cultures and are a more clinically relevant model. A discrete screening of genetically diverse marine samples in the spheroid assay led to the identification of a novel activity for the known compound furospinulosin 1. This compound shows activity against MDA-MB-231 triple negative breast cancer cells grown as spheroids and treated for 24 or 48 h. No cytotoxicity was seen in traditional two-dimensional adherent cultures treated for a longer time (72 h). A reverse phase protein array (RPPA) confirmed the limited activity of the compound in cells grown traditionally and revealed changes in protein expression when cells are grown as spheroids that are associated with better clinical prognosis. Analysis of the RPPA data through the Broad institute’s connectivity map suggested the hypothesis that furospinulosin 1 functions as an MEK inhibitor. Analysis of the RPPA data through STRING supports the apoptosis observed. The selectivity exhibited by furospinulosin 1 for triple negative breast cancer cells only when grown as spheroids makes it an interesting compound with strong therapeutic potential that merits further study.     

Effect of Sorghum Decortication and Use of Protease Before Liquefaction with Thermoresistant α‐Amylase on Efficiency of Bioethanol Production

The aim was to study the dual effect of sorghum decortication and protease treatment before liquefaction with α‐amylase on the performance of subsequent steps of saccharification and fermentation. A bifactorial experiment with a level of confidence of P < 0.05 was designed to study differences among grains (maize, whole, and decorticated sorghum) and the effectiveness of the protease before liquefaction. Sorghum was decorticated to remove most of the pericarp and part of the germ and increase starch concentration of the feedstock. The decorticated sorghum had significantly higher starch hydrolysis during liquefaction compared with the whole kernel. These hydrolyzates contained ≈50% more reducing sugars than the untreated counterparts. At the end of saccharification, the final glucose concentration in hydrolyzates treated without protease was the highest for maize (180 mg/mL), followed by decorticated sorghum (165 mg/mL), and whole sorghum (145 mg/mL). Decortication and protease treatment had a significant effect on fermentation times. In decorticated sorghum mash treated with and without protease, fermentation times were 22 and 60 hr, respectively. The decorticated sorghum treated with protease yielded similar amounts of ethanol compared with maize and 44% more ethanol compared with the untreated whole sorghum. Both sorghum decortication and protease treatments before hydrolysis with α‐amylase are recommended to increase ethanol yields, lower yields of distilled grains, and save liquefaction, saccharification, and fermentation times.     

Effect of seasons and larval food plants on the quality of <Emphasis Type="Italic">Gonometa postica</Emphasis> cocoons

The effect of food plants [Acacia elatior Brenan, A. tortilis (Forssk.) and A. nilotica (L.) Del.] on the development time of Gonometa postica Walker (Lepidoptera: Lasiocampidae) larvae and the quality of their cocoons was studied in the Imba and Mumoni forests of Mwingi, Eastern Kenya, during the long (March-May) and short (October-December) rainy seasons of 2006 and 2007. Larvae were reared in semi-captivity in net sleeves attached to branches of the plants. The period between hatching of eggs and spinning of cocoons, as well as their weight, as determinants of cocoon quality were recorded. In addition, collections of cocoons from the wild were done in the two forests from the same host plants for assessment of their quality. In the cage experiment, larval development period and quality of cocoons differed according to food plants, seasons and sites, whereas quality of cocoons sampled from the wild habitat was similar for all food plants and seasons but varied according to site. Generally, there was a positive correlation between weight of cocoon, its length and width for the two seasons for both the semi-captive population and those from the wild. Larvae reared on A. elatior had a shorter development period and higher cocoon quality than those raised on A. tortilis and A. nilotica. Generally, temperature and relative humidity significantly influenced larval development time and the effect was both positive and negative. Rainfall was generally higher in Mumoni than in Imba. The implications of the above findings for the semi-captive rearing of G. postica to increase the quality of cocoons are discussed.     

Economic performance of the ‘push–pull’ technology for stemborer and Striga control in smallholder farming systems in western Kenya

The ‘push–pull’ technology (PPT), developed in Africa, offers effective control of cereal stemborers and Striga weed in maize-based cropping systems. It involves intercropping maize with desmodium, Desmodium uncinatum, with Napier grass, Pennisetum purpureum, planted as border around this intercrop. Desmodium repels the stemborer moths (push) that are subsequently attracted to the Napier grass (pull). Desmodium also suppresses and eliminates Striga. We assessed economic performance of this technology compared to the conventional maize mono- and maize–bean intercropping systems in six districts in western Kenya over 4–7 years. Ten farmers were randomly recruited in each district and each planted three plots representing the three cropping systems. The cost–benefit analyses were carried out, together with the systems’ net returns to land and labour and their discounted net present values (NPV). Maize grain yields and associated gross margins from the PPT system were significantly higher than those in the other two systems. Although the production costs were significantly higher in the PPT than in the two cropping systems in the first cropping year, these reduced to either the same level or significantly lower than in the maize–bean intercrop from the second year onwards in most of the districts. Similarly, the net returns to land and labour with the PPT were significantly higher than with the other two systems. The PPT consistently produced positive NPV when the incremental flows of its benefits compared to those of the two conventional systems were discounted at 10–30%, indicating that PPT is more profitable than the other two systems under realistic production assumptions. PPT is thus a viable option for enhancing productivity and diversification for smallholder farmers who largely depend on limited land resource. Hence, enhancing farmers’ access to less costly planting materials and promoting quality education and training in the use of this knowledge-intensive technology could stimulate its successful adoption.     

Colloidal hard-sphere crystal growth frustrated by large spherical impurities

Impurities affect the nucleation, growth, and structure of crystals. Here we report the effect of large, spherical, polymethylmethacrylate impurities on the crystal growth of monodisperse, hard, polymethylmethacrylate colloids in a density- and optically matching apolar solvent mixture. Crystal growth, initiated at the bottom of the sample, was studied by imaging sequences of two-dimensional xy slices in the plane of the impurity's center with a laser scanning confocal microscope. Impurities form the center of grain boundaries, and a single fluid particle layer around the impurity persists in all cases. The growth rate sensitively depends on the impurity's size. Crystal growth is inhibited to a greater extent near smaller impurities, pointing to local crystal frustration induced by the curvature of the impurity.     

Fermentative metabolism is induced by inhibiting different enzymes of the branched-chain amino acid biosynthesis pathway in pea plants

The inhibition of branched-chain amino acid (BCAA) biosynthesis was evaluated in pea plants in relation to the ability for induction of fermentative metabolism under aerobic conditions. Chlorsulfuron and imazethapyr (inhibitors of acetolactate synthase, ALS, EC 4.1.3.18) produced a strong induction of pyruvate decarboxylase (PDC, EC 4.1.1.1) and alcohol dehydrogenase (ADH, EC 1.1.1.1) activities and a lesser induction of lactate dehydrogenase (LDH, EC 1.1.1.27) and alanine aminotransferase (AlaAT, EC 2.6.1.2) activities in roots. Inhibition of the second enzyme of the BCAA biosynthesis (ketol-acid reductoisomerase, KARI, EC 1.1.1.86) by Hoe 704 (2-dimethylphosphinoyl-2-hydroxyacetic acid) and CPCA (1,1-cyclopropanedicarboxylic acid) enhanced fermentative enzyme activities including PDC, ADH, and AlaAT. Fermentative metabolism induction occurring with ALS- and KARI-inhibitors was related to a higher expression of PDC. In the case of KARI inhibition, it is proposed that fermentation induction is due to an inhibition of ALS activity resulted from an increase in acetolactate concentration. Fermentative metabolism induction in roots, or at least ethanolic fermentation, appeared to be a general physiological response to the BCAA biosynthesis inhibition.     

Canopy species composition drives seasonal soil characteristics in a Mediterranean riparian forest

European Journal of Forest Research - Mediterranean riparian ecosystems provide key ecosystem services, such as climate regulation, water quality, and flood protection. Forest degradation and...