Decline in glutathione peroxidase and cytoplasmic catalases by lindane may cause an increase of reactive oxygen species in Saccharomyces cerevisiae

Lindane or gamma 1α,2α,3β,4α,5α,6β-hexachlorocyclohexane is an organochlorine insecticide, persistent in soils and aquifers, lipophilic, chemically and biochemically inert that accumulates along the human food chain. It is commonly used on a wide variety of crops, in warehouses, in public health to control insect-borne diseases, and (with fungicides) as a seed treatment. Lindane is also presently used in lotions, creams, and shampoos for the control of lice and mites (scabies) in humans. Several chemicals as lindane, toxic for aquatic organisms, birds and mammals have been in the news recently, since the European Union intend to ban it. Therefore it is urgent to clarify the toxicological mechanisms of this compound in eukaryotic cells. Thus the main purpose of this work was to evaluate the effects of lindane in the wine wild-type Saccharomyces cerevisiae UE-ME3 of Alentejo musts, Portugal, a unicellular eukaryotic organism, described as resistant to the presence of pesticides or metals. Cells at mid-exponential phase were inoculated in YEPD medium with 2 % (w/v) glucose and incubated during 72 h in a water bath with orbital shaking, at 28 ºC, in the absence or in presence of 5 and 50 μM lindane. Samples from each treatment were used to obtain growth curves, wet weight and to prepare post-12000 g supernatant, used for determination of reactive oxygen species (ROS) [1] by fluorimetry and alkaline phosphatase (ALP) [2], glutathione peroxidase (GPx) [3], selenium-dependent glutathione peroxidase (Se-GPx) [3] and cytoplasmic catalase (CAT T) [4] activities as well as pellet for determination of peroxisomal catalase (CAT A) [4] activities by spectrophotometry. The results show that lindane inhibited cell growth of S. cerevisiae UE-ME3, causing a decrease in the biomass produced along 72 h, as well as cell viability from 24 h of assay. On the other hand, was detected an increase in the ROS content of post-12,000 g sediment of cells exposed to 5 μM lindane and post-12000 g supernatant of cells subjected to any exposure conditions, eventually conditioned by a decline in GPx and CAT T activities, which has become the detoxification of hydrogen peroxide less effective. The increase in the CAT A activity without significant changes in the ALP and Se-GPx activities justified, in part, the increase in ROS levels of S. cerevisiae exposed to lindane, as well as the loss of cell viability due to inadequate response of glutathione cycle or cells signaling pathways that assure lipid biosynthesis

Optical-NIR spectroscopy of the puzzling gamma-ray source 3FGL 1603.9-4903/PMN J1603-4904 with X-shooter

The Fermi/LAT instrument has detected about two thousands Extragalactic High Energy (E > 100 MeV) gamma-ray sources. One of the brightest is 3FGL 1603.9-4903, associated to the radio source PMN J1603-4904. Its nature is not yet clear, it could be either a very peculiar BL Lac or a CSO (Compact Symmetric Object) radio source, considered as the early stage of a radio galaxy. The latter, if confirmed, would be the first detection in gamma-rays for this class of objects. Recently a redshift z=0.18 +/- 0.01 has been claimed on the basis of the detection of a single X-ray line at 5.44 +/- 0.05 keV interpreted as a 6.4 keV (rest frame) fluorescent line. We aim to investigate the nature of 3FGL 1603.9-4903/PMN J1603-4904 using optical to NIR spectroscopy. We observed PMN J1603-4904 with the UV-NIR VLT/X-shooter spectrograph for two hours. We extracted spectra in the VIS and NIR range that we calibrated in flux and corrected for telluric absorption and we systematically searched for absorption and emission features. The source was detected starting from ~6300 Ang down to 24000 Ang with an intensity comparable to the one of its 2MASS counterpart and a mostly featureless spectrum. The continuum lacks absorption features and thus is non-stellar in origin and likely non-thermal. On top of this spectrum we detected three emission lines that we interpret as the Halpha-[NII] complex, the [SII] 6716,6731 doublet and the [SIII] 9530 line, obtaining a redshift estimate of z= 0.2321 +/- 0.0004. The equivalent width of the Halpha-[NII] complex implies that PMN J1603-4904 does not follow the observational definition of BL Lac, the line ratios suggest that a LINER/Seyfert nucleus is powering the emission. This new redshift measurement implies that the X-ray line previously detected should be interpreted as a 6.7 keV line which is very peculiar.Comment: Published in Astronomy and Astrophysic

Comparative study of toxicological effects of lindane and isoproturon pesticides in the Saccharomyces cerevisiae

Organochlorine insecticides and phenylurea herbicides such as lindane (hexachlorocyclo-hexane, γ-HCH) and isoproturon (3-(4-isopropylphenyl)-1,1-dimethylurea, IPU), used in agricultural applications for the pest and broad-leaved control, are often found in contaminated groundwater and surface water. Unfortunately, the toxicity of these pesticides in eukaryotic cells is still poorly understood. Saccharomyces cerevisiae is a promising eukaryotic organism for the toxicological evaluation of pollutants, because its metabolism is similar to that of high-level organisms. Thus the aim of this study was to compare the effects of two pesticides on yeast-cell viability and its antioxidant power. S. cerevisiae UE-ME3, a wild-type strain belonging to Oenology Laboratory of the University of Évora, grown in the presence of 50 μM γ-HCH and 100 μM IPU in 2% glucose and peptone deprivation medium (YED), at 28 °C were compared with control cells. CFU were determined at the end of the experiment and remaining cells disintegrated in 10 mM phosphate buffer pH 7.0 by sonication. Post-12,000 × g supernatant was used for determination of glutathione (GSH), glutathione disulphide (GSSG) and malondialdehyde (MDA) content and glutathione reductase (GR), glutathione peroxidase (GPx) and cytoplasmic catalase (CTT1) activity. The results showed that 50 μM γ-HCH exposures, in YED medium, did not affect cell viability, CTT1 activity, non-protein thiols (GSH + GSSG) and MDA content of S. cerevisiae. However, the 50 μM γ-HCH exposure caused a decrease in GSH/GSSG ratio, GR and GPx activity. In contrast, 100 μM IPU exposures caused an increase in the cell viability, (GSH + GSSG), GSH/GSSG ratio and GR activity. Additionally, there was a decrease in the MDA levels, GPx and CTT1 activity, under the same growth conditions. In conclusion, the presence of lindane, in YED medium, caused a reducing-oxidizing transition, a slowdown of the glutathione redox cycle without disturbing the survival of S. cerevisiae. However, the increase in the cell viability, the redox buffer power and the lipid peroxidation attenuation caused by IPU in nitrogen deprivation, appears to be due to the high regeneration capacity of GSH via GR activity, pointing out a possible use of isoproturon as nitrogen source

Chondroprotective Effect of Kartogenin on CD44-Mediated Functions in Articular Cartilage and Chondrocytes

OBJECTIVE: A recent report identified the small molecule kartogenin as a chondrogenic and chondroprotective agent. Since changes in hyaluronan metabolism occur during cartilage degeneration in osteoarthritis, we began studies to determine whether there was a connection between extracellular hyaluronan, CD44–hyaluronan interactions and the effects of kartogenin on articular chondrocytes. METHODS: Chondrocytes cultured in monolayers, bioengineered neocartilages, or cartilage explants were treated with kartogenin with or without stimulation by IL-1β. Accumulation of matrix was visualized by a particle exclusion assay or by safranin O staining and release of sulfated glycosaminoglycans was determined. Production of aggrecanases and aggrecan G1-ITEGE neoepitope, fragmentation of CD44 and the SMAD1/5/8 signaling pathway were evaluated by western blotting. RESULTS: Kartogenin treatment enhanced chondrocyte pericellular matrix assembly and retention in the presence of IL-1β. The chondroprotective effects of kartogenin on IL-1β-induced release of sulfated glycosaminoglycans from articular cartilage explants, reduction in safranin O staining of neocartilage discs as well as a reduction in aggrecan G1-ITEGE neoepitope in chondrocyte and explant cartilage cultures were observed. Kartogenin partially blocked the IL-1β-induced increased expression of ADAMTS-5. Additionally, kartogenin-treated articular chondrocytes exhibited a decrease in CD44 proteolytic fragmentation. However, kartogenin treatment did not enhance proteoglycan in control, non-IL-1β-treated cultures. Similarly, kartogenin enhanced the SMAD1 phosphorylation but only following pretreatment with IL-1β. CONCLUSION: These studies provide novel information on the chondroprotective function of kartogenin in adult articular cartilage. The effects of kartogenin are significant after activation of chondrocytic chondrolysis, which may occur following disruption of homeostasis maintained by hyaluronan–CD44 interactions

Assessing the severity and the incidence of Cassava Root Necrosis Disease (CRND) in western Democratic Republic of Congo

Open Access ArticleCassava is the staple food in the Democratic Republic of Congo (DRC) where both the roots and leaves are consumed. This crop is susceptible to several viral diseases, including Cassava Mosaic Disease(CMD) and Cassava Brown Streak Disease(CBSD) in eastern DRC. Following earlier studies that show root necrosis occurring in western DR Care not due to CBSD but to Cassava Root Necrosis Disease (CRND), an exploratory survey was conducted in western DRC from 2016 to 2017 in order to determine the distribution, the severity and the incidence of this disease (previously known as CBSD-like disease). NGS ( Next Generation Sequencing) results confirmed all the previous negative results obtained using PCR and CBSV primers. This suggests that microorganisms such as bacteria or fungi could be responsible for cassava root necrosis in western DRC and is not CBSD as predicted. Five provinces (Bas-Congo, Kinshasa, Bandundu, Equateur and Kasai-Oriental) were surveyed and data were collected according to the harmonized protocols adopted by countries within the West African Virus Epidemiology (WAVE) project. Statistical tests (ANOVA) performed on our data showed that CRND severity did not vary significantly among the provinces of Kinshasa, Bandundu and Bas-Congo which are the areas most affected by the disease. Bas-Congo and Kinshasa provinces presented the highest maximum disease severity (score 3 and 5 respectively), while Equateur province had the lowest disease severity score. Equateur province also had the highest percentage of healthy plants and few plants presented mild symptoms. The overall average of cassava root necrosis severity in western DRC ranged around 1.88 ± 0.08, an approximate score of 2. The overall mean incidence of CRND in western DRC was 22.24 ± 2.4% but reached 100% in localities considered as hotspots (Lukuakua in Bas-Congo and Nguma in Plateau des Batékés). The behaviour of cassava varieties against CRND is similar with CBSD in East Africa, most of improved varieties and landraces are susceptible to both diseases. Correlation analyses showed a positive correlation (r = 0.6940) between severity and incidence of CRND. Therefore, Bas-Congo province is the most affected province, while the province of Equateur is the least affected province in western DRC. Further investigations, including genomic surveillance, should also be conducted in the eastern DRC where CBSD is confirmed to know if CRND is found in conjunction with CBSD and to report possible instances of mixed infections. For medium-term disease control, our study suggests that the development and deployment of control measures including cultivars with resistance to CRND and CBSD should be a priority

Modulation and Signal Processing for LEO-LEO Optical Inter-satellite Links

We investigate key aspects of coherent optical communications on inter-satellite links (ISLs) for the next-generation ultra-dense low-Earth orbit (LEO) constellations. Initially, the suitability of QPSK, 8-QAM, and 16-QAM modulation formats with different symbol rates (28 GBaud, 60 GBaud, and 120 GBaud) and channel coding schemes (oFEC and staircase codes) for intra- and interorbital connections is evaluated. We provide SNR margins for all investigated sets and determine unfeasible operating points. We show that sets with higher-order modulation formats combined with high symbol rates can prove unfeasible, even for first-neighbor connections. Furthermore, the presence or absence of optical pre-amplification as well as the choice for a more robust channel coding technique, such as the oFEC, can be decisive in certain LEO-LEO links. Next, we characterize the Doppler shift (DS) and its time derivative for first-neighbor interorbital connections in two different topologies and for general connections established between any pairs of satellites. Our results reveal that while the maximum Doppler-generated frequency shift amplitude can be considerably higher than those typically found in fiber-optic communications, the time derivative values are significantly lower. Finally, we address all-digital DS compensation in extreme cases of frequency offset amplitude and derivative where the typical Mth-power algorithm is not sufficient. To this end, we propose a filtered version of an existing two-stage method combining spectral shifts with the Mth-power method. The simulation results indicate that this approach provides an appropriate solution for all examined cases.Comment: 13 pages, 7 figures, 6 table

First report and preliminary evaluation of cassava root necrosis in Angola

Open Access ArticleCassava is a main staple food for 800 million people world-wide. Production is limited by pest and pathogens. The most devastating cassava viruses are Cassava Brown Streak Virus and Uganda Cassava Brown Streak Virusboth causing severe root necrosis called Cassava Brown Streak Disease. In the last 10 years, the Cassava Brown Streak Disease (CBSD)has spread across Africa from the east coast of Africa to central Africa. Similar root necrosis to cassava brown streak disease has also been identified in the Democratic Republic of Congo where the first symptoms were identified in 2002 in Kinshasa and Kongo central province. In 2012, the presence of CBSD was confirmed in eastern Democratic Republic of Congo. All attempts since 2002 in western Democratic Republic of Congo to identify the cause of these root necrosis have failed. In 2017, a team of scientists surveying the Songololo Territory in the Kongo central province at the northern Angola, identified the same root necrosis similar to CBSD in several localities bordering Angola. These unexpected results will foreshadow the presence of cassava root necrosis in Angola. This preliminary investigation in northern Angola was conducted specifically in the Zaire province and the territory of Mbanza Kongo at approximatively 62 kms from the Democratic Republic of Congo border in order to verify, whether or not, these root necrosis are present in Angola. Results obtained from this exploratory survey in several fields of the Zaire province and territory of Mbanza Kongo confirmed, for the first time, the presence of cassava root necrosis in Angola, similar to CBSD, as identified in western DRC

Avaliação do efeito do corte horizontal da copa, com máquina de podar de discos, em pomares de pera “Rocha

Current pear pruning making use of pneumatic shears still is a very labour intensive operation. The Proder project “Avaliação da poda mecânica em pomares de pera” was designed to contribute to solutions that would reduce the present dependence in labour and therefore to promote a reduction in pruning costs. This paper shows the results of a trial made to evaluate the influence of mechanical topping in manual pruning complement field work and pear yield. Topping was performed using a Reynolds 6DT 3.0m cutting bar with six hydraulic-driven circular disc-saws mounted in the three point tractor linkage system. The field trial was performed in a commercial orchard with 20 years, planted in an array of 4m x 2m with tree lines oriented in North-South direction. Trees were trained as the central leader system. In this trial, in a randomised complete block design with four replications, two treatments are being compared leading to 8 plots with one line of 14 trees per plot. The treatments tests were: T1 - manual pruning performed by workers using pneumatic shears, in each year; T2 - Topping the canopy parallel to the ground, using a discs-saw pruning machine mounted in a front loader of an agricultural tractor, followed by manual pruning complement performed by workers with pneumatic shears. Tree height and width was measured, before and after pruning. Work was timed and pear yields evaluated. Mechanical topping seems to be effective in the control of tree height, which can contribute to increase 14% of work rates on manual pruning complement. No significant differences in pear yield were found between treatments