Biochemical basis of organophosphate and carbamate resistance in Asian citrus psyllid

The Asian citrus psyllid, Diaphorina citri Kuwayama, is a worldwide pest of citrus, which vectors the putative causal pathogen of huanglongbing. Current management practices warrant continuous monitoring of field populations for insecticide resistance. Baseline activities of acetylcholinesterase (AChE), general esterase, and glutathione S-transferase as well as sensitivity of AChE to selected organophosphate and carbamate insecticides were established for a susceptible laboratory strain (Lab) and compared with several field populations of D. citri from Florida. The specific activity of AChE in various D. citri populations ranged from 0.77 to 1.29 microM min(-1) mg of protein(-1); the Lab strain was characterized by the highest activity. Although reduced AChE sensitivity was observed in the Lab strain compared with field populations, overlap of 95% confidence intervals of I50 values (concentration required for 50% AChE activity inhibition) suggests no significant difference in AChE sensitivity among all populations tested for a given insecticide. There was no significant evidence of target site insensitivity in field populations that were exposed to the selected organophosphate and carbamate insecticides tested. The specific activity of general esterase and glutathione S-transferase was lowest in the Lab strain and was generally comparable to that of the field populations evaluated. The current data provide a mode-of-action specific baseline for future monitoring of resistance to organophosphate and carbamate insecticides in populations of D. citri.     

Impact of low and high fluence rates of UV-B radiation on growth and oxidative stress in Phormidium foveolarum and Nostoc muscorum under copper toxicity: differential display of antioxidants system

In the present study, impact of low (UV-B_L) and high (UV-B_H) fluence rates of UV-B on growth, oxidative stress and antioxidant system was studied in two cyanobacteria i.e. Phormidium foveolarum and Nostoc muscorum under Cu (2 and 5???M) toxicity after 24 and 72?h of experiments. UV-B_H and Cu treatment decreased growth of both the cyanobacteria and Cu induced decrease in growth was accompanied by a significant increase in Cu accumulation. Levels of reactive oxygen species (ROS), i.e. superoxide radicals (SOR; $ \text O_{2}^{\cdot\,-} $ ) and hydrogen peroxide (H₂O₂) were significantly increased by Cu and UV-B_H exposure which in turn accelerated lipid peroxidation (malondialdehyde: MDA) and protein oxidation (reactive carbonyl groups: RCG). Activities of enzymatic antioxidants, such as superoxide dismutase (SOD), peroxidase (POD) and glutathione-S-transferase (GST) were increased by both doses of Cu as well as UV-B. Conversely, Cu and UV-B_H drastically decreased catalase (CAT) activity. After the commencement of 24?h of treatment with Cu alone and together with UV-B_H, non-protein thiols (NP-SH) contents were decreased while after 72?h, a reverse trend was noticed. Unlike NP-SH, cysteine content decreased appreciably during the treatments. In contrast to this, low dose (UV-B_L) of UV-B did not influence growth, SOR, H₂O₂, MDA and RCG contents. An improvement in CAT activity and NP-SH content was observed under Cu and UV-B_L treatment; hence, UV-B_L treatment resulted into certain degree of protection against Cu toxicity in both the organisms. Thus, the results showed that UV-B_H and UV-B_L exerted differential effects on both the organisms under Cu toxicity, and compared to N. muscorum, P. foveolarum was less affected by Cu and UV-B_H.     

Impact of exogenous silicon addition on chromium uptake, growth, mineral elements, oxidative stress, antioxidant capacity, and leaf and root structures in rice seedlings exposed to hexavalent chromium

Hydroponic experiments were conducted to investigate the role of exogenous silicon (Si) addition in increasing hexavalent chromium (Cr VI) tolerance in rice seedlings. Rice seedlings were grown under 100 μM Cr(VI) stress without or with 10 μM Si. Chromium treatment decreased growth, photosynthetic pigments and protein, which was accompanied by a significant increase in Cr accumulation and lipid peroxidation (as malondialdehyde; MDA). However, Si addition alleviated Cr toxicity and promoted growth of rice by decreasing Cr accumulation, root-to-shoot Cr transport and MDA level. Contents of macro (Mg, Ca and K) as well as micronutrients (Zn and Fe) were decreased by Cr except Mn while Si addition prevented decrease in these nutrients induced by Cr. Antioxidant capacity and total phenolic contents were decreased by Cr while these indices improved by Si addition. Treatment of Cr decreased the length of leaf epidermal cells and stomatal frequency, and adversely affected chloroplasts containing mesophyll cells and integrity of xylem and phloem, and Si addition minimized these abnormalities. However, frequency of root hairs was increased by Cr treatment. Results showed that exogenous Si addition enhanced Cr(VI) tolerance in rice seedlings by decreasing Cr accumulation, root-to-shoot Cr transport and MDA level, and by increasing content of some mineral elements (K, Fe and Zn) and antioxidant capacity compared to the Cr treatment alone.     

Scaffold hybridization in generation of indenoindolones as anticancer agents that induce apoptosis with cell cycle arrest at G2/M phase

Scaffold hybridization of several natural and synthetic anticancer leads led to the consideration of indenoindolones as potential novel anticancer agents. A series of these compounds were prepared by a diversity-feasible synthetic method. They were found to possess anticancer activities with higher potency compared to etoposide and 5-fluorouracil in kidney cancer cells (HEK 293) and low toxicity to corresponding normal cells (Vero). They exerted apoptotic effect with blocking of cell cycle at G2/M phase.     

Synthesis and antileukemic activity of 16E-[4-(2-carboxy)ethoxybenzylidene]-androstene amides

In order to determine the structural requirements for cytotoxicity against various tumor cell lines, a new series of 16E-arylidene androstene amides with varying degrees of unsaturation in ring A has been synthesized. Characterization and invitro cytotoxic studies of the newly synthesized compounds are discussed. The compounds on evaluation against various tumor cell lines exhibited significant growth inhibition on leukemia cell lines. 3-Chloro-16E-{[4-(4-methylpiperazin-1-yl)-2-oxoethoxy]benzylidene}androst-5-en-17-one (10) emerged as the most potent compound of the series with GI(50) values of 3.94, 2.61, 6.90 and 1.79μM against CCRF-CEM, K-562, RPMI-8226 and SR leukemia cell lines, respectively.     

Membrane association of the PTEN tumor suppressor: molecular details of the protein-membrane complex from SPR binding studies and neutron reflection

The structure and function of the PTEN phosphatase is investigated by studying its membrane affinity and localization on in-plane fluid, thermally disordered synthetic membrane models. The membrane association of the protein depends strongly on membrane composition, where phosphatidylserine (PS) and phosphatidylinositol diphosphate (PI(4,5)P₂) act pronouncedly synergistic in pulling the enzyme to the membrane surface. The equilibrium dissociation constants for the binding of wild type (wt) PTEN to PS and PI(4,5)P₂ were determined to be K_d∼12 µM and 0.4 µM, respectively, and K_d∼50 nM if both lipids are present. Membrane affinities depend critically on membrane fluidity, which suggests multiple binding sites on the protein for PI(4,5)P₂. The PTEN mutations C124S and H93R show binding affinities that deviate strongly from those measured for the wt protein. Both mutants bind PS more strongly than wt PTEN. While C124S PTEN has at least the same affinity to PI(4,5)P₂ and an increased apparent affinity to PI(3,4,5)P₃, due to its lack of catalytic activity, H93R PTEN shows a decreased affinity to PI(4,5)P₂ and no synergy in its binding with PS and PI(4,5)P₂. Neutron reflection measurements show that the PTEN phosphatase “scoots" along the membrane surface (penetration <5 Å) but binds the membrane tightly with its two major domains, the C2 and phosphatase domains, as suggested by the crystal structure. The regulatory C-terminal tail is most likely displaced from the membrane and organized on the far side of the protein, ∼60 Å away from the bilayer surface, in a rather compact structure. The combination of binding studies and neutron reflection allows us to distinguish between PTEN mutant proteins and ultimately may identify the structural features required for membrane binding and activation of PTEN.     

Artemisinin resistance containment project in Thailand. i: implementation of electronic-based malaria information system for early case detection and individual case management in provinces along the Thai-Cambodian border

ABSTRACT: BACKGROUND: The Bureau of Vector-borne Diseases, Ministry of Public Health, Thailand, has implemented an electronic Malaria Information System (eMIS) as part of a strategy to contain artemisinin resistance. The attempt corresponds to the WHO initiative, funded by the Bill & Melinda Gates Foundation, to contain anti-malarial drug resistance in Southeast Asia. The main objective of this study was to demonstrate the eMIS' functionality and outputs after implementation for use in the Thailand artemisinin-resistance containment project. METHODS: The eMIS had been functioning since 2009 in seven Thai-Cambodian border provinces. The eMIS has covered 61 malaria posts/clinics, 27 Vector-borne Disease Units covering 12,508 hamlets at risk of malaria infections. The eMIS was designed as an evidence-based and near real-time system to capture data for early case detection, intensive case investigation, monitoring drug compliance and on/off-site tracking of malarial patients, as well as collecting data indicating potential drug resistance among patients. Data captured by the eMIS in 2008-2011 were extracted and presented. RESULTS: The core functionalities of the eMIS have been utilized by malaria staff at all levels, from local operational units to ministerial management. The eMIS case detection module suggested decreasing trends during 2009-2011; the number of malaria cases detected in the project areas over the years studied were 3818, 2695, and 2566, with sero-positive rates of 1.24, 0.98, and 1.16%, respectively. The eMIS case investigation module revealed different trends in weekly Plasmodium falciparum case numbers, when classified by responsible operational unit, local and migrant status, and case-detection type. It was shown that most Thai patients were infected within their own residential district, while migrants were infected either at their working village or from across the border. The data mapped in the system suggested that P. falciparum-infected cases and potential drug-resistant cases were scattered mostly along the border villages. The mobile technology application has detected different follow-up rates, with particularly low rates among seasonal and cross-border migrants. CONCLUSION: The eMIS demonstrated that it could capture essential data from individual malaria cases at local operational units, while effectively being used for situation and trend analysis at upper-management levels. The system provides evidence-based information that could contribute to the control and containment of resistant parasites. Currently, the eMIS is expanding beyond the Thai-Cambodian project areas to the provinces that lie along the Thai-Myanmar border.     

The influence of light on microtubule dynamics and alignment in the Arabidopsis hypocotyl

Light and dark have antagonistic effects on shoot elongation, but little is known about how these effects are translated into changes of shape. Here we provide genetic evidence that the light/gibberellin-signaling pathway affects the properties of microtubules required to reorient growth. To follow microtubule dynamics for hours without triggering photomorphogenic inhibition of growth, we used Arabidopsis thaliana light mutants in the gibberellic acid/DELLA pathway. Particle velocimetry was used to map the mass movement of microtubule plus ends, providing new insight into the way that microtubules switch between orthogonal axes upon the onset of growth. Longitudinal microtubules are known to signal growth cessation, but we observed that cells also self-organize a strikingly bipolarized longitudinal array before bursts of growth. This gives way to a radial microtubule star that, far from being a random array, seems to be a key transitional step to the transverse array, forecasting the faster elongation that follows. Computational modeling provides mechanistic insight into these transitions. In the faster-growing mutants, the microtubules were found to have faster polymerization rates and to undergo faster reorientations. This suggests a mechanism in which the light-signaling pathway modifies the dynamics of microtubules and their ability to switch between orthogonal axes.     

Fast Carbon Footprinting for Large Product Portfolios

Publicly Available Specification 2050‐2011 (PAS 2050), the Green House Gas Product Protocol (GHGPP) standard and forthcoming guideline 14067 from the International Organization for Standardization (ISO) have helped to propel carbon footprinting from a subdiscipline of life cycle assessment (LCA) to the mainstream. However, application of carbon footprinting to large portfolios of many distinct products and services is immensely resource intensive. Even if achieved, it often fails to inform company‐wide carbon reduction strategies because footprint data are disjointed or don't cover the whole portfolio. We introduce a novel approach to generate standard‐compliant product carbon footprints (CFs) for companies with large portfolios at a fraction of previously required time and expertise. The approach was developed and validated on an LCA dataset covering 1,137 individual products from a global packaged consumer goods company. Three novel techniques work in concert in a single approach that enables practitioners to calculate thousands of footprints virtually simultaneously: (i) a uniform data structure enables footprinting all products and services by looping the same algorithm; (ii) concurrent uncertainty analysis guides practitioners to gradually improve the accuracy of only those data that materially impact the results; and (iii) a predictive model generates estimated emission factors (EFs) for materials, thereby eliminating the manual mapping of a product or service's inventory to EF databases. These autogenerated EFs enable non‐LCA experts to calculate approximate CFs and alleviate resource constraints for companies embarking on large‐scale product carbon footprinting. We discuss implementation roadmaps for companies, including further road‐testing required to evaluate the effectiveness of the approach for other product portfolios, limitations, and future improvements of the fast footprinting methodology.