Perturbing Circadian Oscillations in an In Vitro Suprachiasmatic Nucleus With Magnetic Stimulation

Many neurological disorders are associated with abnormal oscillatory dynamics. The suprachiasmatic nucleus (SCN) is responsible for the timing and synchronization of physiological processes. We performed experiments on PERIOD2::LUCIFERASE transgenic “knock-in” mice. In these mice, a gene that is expressed in a circadian pattern is fused to an inserted gene that codes for luciferase, which is a bioluminescent enzyme. A one-time 3 min magnetic stimulation (MS) was applied to excised slices of the SCN. The MS consisted of a 50-mT field that was turned on and off 4,500 times. The rise time and fall time of the field were 75 μs. A photon count that extended over the full 5 days that the slice remained viable, subsequently revealed how the MS affected the circadian cycle. The MS was applied at points in the circadian cycle that correspond to either maximal or minimal bioluminescence. It was found that both the amplitude and period of the endogenous circadian oscillation are affected by MS and that the effects strongly depend on where in the circadian cycle the stimulation was applied. Our MS dose is in the same range as clinically applied doses, and our findings imply that transcranial MS may be instrumental in remedying disorders that originate in circadian rhythm abnormalities. Bioelectromagnetics. 2020;41:63–72 © 2019 Wiley Periodicals, Inc.     

Hybridization of nickel oxide nanoparticles with carbon dots and its application for antibacterial activities

A nickel oxide nanoparticle (NiO NP) composite with carbon dots (C-dots), (NiO NPs@C-dots) was synthesized, characterized, and then its antibacterial activity was evaluated. NiO NPs were prepared using Buddleja polystachya Fresen leaf extract and Ni(NO₃)₂.6H₂O as precursors. The C-dots were synthesized from benzene-1,4-diamine and citric acid. The cubic structure of the NiO NPs and NiO NPs@C-dots was in phase with their average particle size distributions of 21.47 ± 0.56 and 21.61 ± 0.34 nm, respectively. The surface morphology of the NiO NPs@C-dots was characterized using field emission scanning electron microscopy and also revealed a large surface area, which is advantageous for the specified application. The X-ray diffraction result indicated a cubic face wurtzite structure and the crystalline nature of the NiO NPs. Carbon-doped compounds had no influence on the crystal structure of the NiO compound and no new peaks were observed. The antibacterial activity of a composite made up of NiO NPs@C-dots was tested, as well as the antibacterial activities of compounds produced against human photogenic bacterial strains. Both NiO NPs and NiO NPs@C-dots were found to be powerful against all bacterial strains, based on the bioassay results. NiO NPs and NiO@C-dots appeared to display strong to inhibitory effects of 14–20 mm and 17–23 mm, respectively.     

Seasonal variation in primary production of a large high altitude tropical lake (Lake Tana,Ethiopia): effects of nutrient availability and water transparency

Primary production rates, chlorophyll and phytoplankton biovolume were measured monthly from April 2003 to November 2004 in Lake Tana, a large tropical lake in the highlands of Ethiopia. The lake is characterised by low nutrient concentrations, and a low water transparency due to high silt load of the inflowing rivers during the rainy seasons (May–November) and daily resuspension of sediments in the inshore zone. The mean chlorophyll-a concentrations varied seasonally and ranged from 2.6 mg m⁻³ to 8.5 mg m⁻³ (mean: 4.5 mg m⁻³) in the offshore zone. Primary production was measured using the light–dark bottles technique. We incubated only at three depths, i.e. 0.6, 1.2 and 1.8 m. Therefore, we may have missed a substantial part of the depth production profile and probably also frequently missed P _max. Gross primary production in the openwater averaged 2.43 g O₂ m⁻² d⁻¹ and ranged between 0.03 g O₂ m⁻² d⁻¹ and 10.2 g O₂ m⁻² d⁻¹; production was significantly higher in the inshore zone. The highest production rates were observed in the post-rainy season (Oct–Nov), which coincided with a bloom of Microcystis and higher chlorophyll levels. This seasonal high production is probably caused by a relatively high nutrient availability in combination with favourable light conditions. The gross primary production rates of L. Tana are among the lowest compared with other tropical lakes. This will be partly the result of our underestimation of gross primary production by often missing P _max. Another cause is the oligotrophic nature of the lake in combination with its relatively low water transparency. The gross primary production per unit chlorophyll in the openwater zone was in the same range as in 30 other tropical lakes and reservoirs. The higher primary production in the inshore zone is probably the result of the daily water column mixing (Z _mix ≥ Z _t) in this area, enhancing nutrient recycling. A large proportion of the annual primary production is realised in one of the four seasons only. This productive post-rainy season is relatively short (2 months) and therefore efficiency of transfer of matter between the first and second trophic level of the Lake ecosystem will be poor.     

Cholesterol-depleting statin drugs protect postmitotically differentiated human neurons against ethanol- and human immunodeficiency virus type 1-induced oxidative stress in vitro

The majority of human immunodeficiency virus type 1 (HIV-1)-infected individuals are either alcoholics or prone to alcoholism. Upon ingestion, alcohol is easily distributed into the various compartments of the body, particularly the brain, by crossing through the blood-brain barrier. Both HIV-1 and alcohol induce oxidative stress, which is considered a precursor for cytotoxic responses. Several reports have suggested that statins exert antioxidant as well as anti-inflammatory pleiotropic effects, besides their inherent cholesterol-depleting potentials. In our studies, postmitotically differentiated neurons were cocultured with HIV-1-infected monocytes, T cells, or their cellular supernatants in the presence of physiological concentrations of alcohol for 72 h. Parallel cultures were pretreated with statins (atorvastatin and simvastatin) with the appropriate controls, i.e., postmitotically differentiated neurons cocultured with uninfected cells and similar cultures treated with alcohol. The oxidative stress responses in the presence/absence of alcohol in these cultures were determined by the production of the well-characterized oxidative stress markers, 8-isoprostane-F2-alpha, total nitrates as an indicator for various isoforms of nitric oxide synthase activity, and heat shock protein 70 (Hsp70). An in vitro culture of postmitotically differentiated neurons with HIV-1-infected monocytes or T cells as well as supernatants from these cells enhanced the release of 8-isoprostane-F2-alpha in the conditioned medium six- to sevenfold (monocytes) and four- to fivefold (T cells). It was also observed that coculturing of HIV-1-infected primary monocytes over a time period of 72 h significantly elevated the release of Hsp70 compared with that of uninfected controls. Cellular supernatants of HIV-1-infected monocytes or T cells slightly increased Hsp70 levels compared to neurons cultured with uninfected monocytes or T-cell supernatants (controls). Ethanol (EtOH) presence further elevated Hsp70 in both infected and uninfected cultures. The amount of total nitrates was significantly elevated in the coculture system when both infected cells and EtOH were present. Surprisingly, pretreatment of postmitotic neurons with clinically available inhibitors of HMG-coenzyme A reductase (statins) inhibited HIV-1-induced release of stress/toxicity-associated parameters, i.e., Hsp70, isoprostanes, and total nitrates from HIV-1-infected cells. The results of this study provide new insights into HIV-1 neuropathogenesis aimed at the development of future HIV-1 therapeutics to eradicate viral reservoirs from the brain.     

Effects of Row-Type,Row-Spacing,Seeding Rate,Soil-Type,and Cultivar Differences on Soybean Seed Nutrition under US Mississippi Delta Conditions

The new Early Soybean Production System (ESPS), developed in the Midsouth USA, including the Mississippi delta, resulted in higher yield under irrigated and non-irrigated conditions. However, information on the effects of the agricultural practices such as row-type (RT: twin- vs. single-row), row-spacing, (RS), seeding rate (SR), soil-type (ST) on seed nutrition under the ESPS environment in the Mississippi delta is very limited. Our previous research in the Mississippi delta showed these agricultural practices altered seed nutrients in one cultivar only. However, whether these effects on seed nutrients will be exhibited by other soybean cultivars with earlier and later maturities across multiple years are not yet known. Therefore, the objective of this research was to evaluate the effects of agricultural practices and cultivar (Cv) differences on seed nutrition in clay and sandy soils under ESPS environment of high heat and drought. Two field experiments were conducted; one experiment was conducted in 2009 and 2010, and the other in 2008, 2009, and 2010 under irrigated conditions. Soybean were grown on 102 cm single-rows and on 25 cm twin-rows with 102 cm centers at seeding rates of 20, 30, 40, and 50 seeds m^–2. Two soybean cultivars (94M80 with earlier maturity; and GP 533 with later maturity) were used. Results showed that increasing seeding rate resulted in increases of protein, sucrose, glucose, raffinose, B, and P concentrations on both single- and twin-rows. However, this increase became either constant or declined at the higher rates (40 and 50 seeds m^–2). Protein and linolenic acid concentrations were higher in GP 533 than in 94M80 on both row-types, but oil and oleic acid concentrations were in 94M80 than GP 533. Generally, cultivar GP 533 accumulated more seed constituents in seeds than 94M80. In 2010, there were no clear responses of seed nutrients to SR increase in both cultivars, perhaps due to drier year and high heat in 2010. It is concluded that RT and SR can alter seed nutrition under clay and sandy soils, especially under high heat and drought conditions as in the Mississippi delta.     

Affordable and robust phenotyping framework to analyse root system architecture of soil‐grown plants

The phenotypic analysis of root system growth is important to inform efforts to enhance plant resource acquisition from soils; however, root phenotyping remains challenging because of the opacity of soil, requiring systems that facilitate root system visibility and image acquisition. Previously reported systems require costly or bespoke materials not available in most countries, where breeders need tools to select varieties best adapted to local soils and field conditions. Here, we report an affordable soil‐based growth (rhizobox) and imaging system to phenotype root development in glasshouses or shelters. All components of the system are made from locally available commodity components, facilitating the adoption of this affordable technology in low‐income countries. The rhizobox is large enough (approximately 6000 cm² of visible soil) to avoid restricting vertical root system growth for most if not all of the life cycle, yet light enough (approximately 21 kg when filled with soil) for routine handling. Support structures and an imaging station, with five cameras covering the whole soil surface, complement the rhizoboxes. Images are acquired via the Phenotiki sensor interface, collected, stitched and analysed. Root system architecture (RSA) parameters are quantified without intervention. The RSAs of a dicot species (Cicer arietinum, chickpea) and a monocot species (Hordeum vulgare, barley), exhibiting contrasting root systems, were analysed. Insights into root system dynamics during vegetative and reproductive stages of the chickpea life cycle were obtained. This affordable system is relevant for efforts in Ethiopia and other low‐ and middle‐income countries to enhance crop yields and climate resilience sustainably.     

Arsenopyrite Weathering and Leaching of Arsenic in an Austrian Soil

The extent of arsenopyrite weathering in relation to co-existing minerals in an Austrian soil and the leaching of arsenic from the soil has been investigated. Soil and underlying bedrock samples were collected and characterized by chemical and mineralogical analyses. The solubility of the soil arsenic under anaerobic conditions was studied by incubating the soil sample in distilled water for different periods of time using a customized lycimeter. The solubility of arsenic from pure arsenopyrite mineral and mixtures of arsenopyrite with chalcopyrite or pyrite was studied by incubating the pulverized minerals. Speciation of arsenic in the incubated and non-incubated soil samples was carried out by sequential leaching, solvent-extraction, and ion exchange chromatographic techniques.

Results of SEM analysis indicated that arsenopyite (FeAsS), the most common mineral in the area, occurs in paragenesis with pyrite (FeS₂) and chalcopyrite (CuFeS₂). The existence of these minerals with arsenopyrite was found to enhance its solubilization. From the speciation study it was found that nearly all (92%) of the arsenic in the soil exists in the inorganic form. Out of the total inorganic arsenic, the trivalent inorganic species accounted for only 3% and the remaining 89% was found to be the pentavalent form. The low solubility of As in the Graz soil is attributed to the prevalence of this pentavalent inorganic species.     

Caenorhabditis elegans NDX-4 is a MutT-type enzyme that contributes to genomic stability

MutT enzymes prevent DNA damage by hydrolysis of 8-oxodGTP, an oxidized substrate for DNA synthesis and antimutagenic, anticarcinogenic, and antineurodegenerative functions of MutT enzymes are well established. MutT has been found in almost all kingdoms of life, including many bacterial species, yeasts, plants and mammals. However, a Caenorhabditis elegans MutT homologue was not previously identified. Here, we demonstrate that NDX-4 exhibits both hallmarks of a MutT-type enzyme with an ability to hydrolyze 8-oxodGTP and suppress the Escherichia coli mutT mutator phenotype. Moreover, we show that NDX-4 contributes to genomic stability in vivo in C. elegans. Phenotypic analyses of an ndx-4 mutant reveal that loss of NDX-4 leads to upregulation of key stress responsive genes that likely compensate for the in vivo role of NDX-4 in protection against deleterious consequences of oxidative stress. This discovery will enable us to use this extremely robust genetic model for further research into the contribution of oxidative DNA damage to phenotypes associated with oxidative stress.