Drought-induced changes in nutrient concentrations and retention in two shallow Mediterranean lakes subjected to different degrees of management

While extensive knowledge exists on the relationship between nutrient loading and nutrient concentrations in lakes in the cold temperate region, few studies have been conducted in warm lakes, not least in warm arid lakes. This is unfortunate as a larger proportion of the world’s lakes will be situated in arid climates in the future due to climate change and a larger proportion will suffer from a higher frequency of intensive drought. We conducted a comprehensive 11–13 year mass balance study in two interconnected shallow Mediterranean lakes in Turkey, covering a period with substantial changes in climate conditions. The upstream lake was only affected by natural changes in nutrient loading, while the downstream lake was additionally influenced by sewage diversion and restoration by fish removal. Contrasting to experience from north temperate lakes we found an increase in in-lake concentrations of total phosphorus and inorganic nitrogen (ammonia as well as nitrate) in dry years despite lower external nutrient loading, and submerged macrophytes did not increase the nitrogen retention capacity of the lakes. In contrast, fish removal modulated the nitrogen concentration as in north temperate lakes, but the effect was not long-lasting. Our results suggest that climate warming reduces the nutrient retention capacity of shallow lakes in the Mediterranean and exacerbates eutrophication. Lower thresholds of nutrient loading for shifting turbid shallow lakes to a clear water state are therefore to be expected in arid zones in a future warmer climate, with important management implications.     

A Centrosome-autonomous Signal That Involves Centriole Disengagement Permits Centrosome Duplication in G2 Phase after DNA Damage

DNA damage can induce centrosome overduplication in a manner that requires G2-to-M checkpoint function, suggesting that genotoxic stress can decouple the centrosome and chromosome cycles. How this happens is unclear. Using live-cell imaging of cells that express fluorescently tagged NEDD1/GCP-WD and proliferating cell nuclear antigen, we found that ionizing radiation (IR)-induced centrosome amplification can occur outside S phase. Analysis of synchronized populations showed that significantly more centrosome amplification occurred after irradiation of G2-enriched populations compared with G1-enriched or asynchronous cells, consistent with G2 phase centrosome amplification. Irradiated and control populations of G2 cells were then fused to test whether centrosome overduplication is allowed through a diffusible stimulatory signal, or the loss of a duplication-inhibiting signal. Irradiated G2/irradiated G2 cell fusions showed significantly higher centrosome amplification levels than irradiated G2/unirradiated G2 fusions. Chicken–human cell fusions demonstrated that centrosome amplification was limited to the irradiated partner. Our finding that only the irradiated centrosome can duplicate supports a model where a centrosome-autonomous inhibitory signal is lost upon irradiation of G2 cells. We observed centriole disengagement after irradiation. Although overexpression of dominant-negative securin did not affect IR-induced centrosome amplification, Plk1 inhibition reduced radiation-induced amplification. Together, our data support centriole disengagement as a licensing signal for DNA damage-induced centrosome amplification.     

Screening of Biological Activities of a Series of Chalcone Derivatives against Human Pathogenic Microorganisms

In an effort to develop new antimicrobial agents, a series of chalcone derivatives, 3 – 60 , were prepared by Claisen–Schmidt condensation of appropriate acetophenones and 2‐furyl methyl ketones with appropriate aromatic aldehydes, furfural, and thiophene‐2‐carbaldehyde in an aqueous solution of NaOH and EtOH at room temperature. The synthesized compounds were characterized by means of their IR‐ and NMR‐spectral data, and elemental analysis. All compounds were tested for their antibacterial and antifungal activities by the disc diffusion method. For the most active compounds, also minimum inhibitory concentrations (MICs) were determined.     

Influence of surgical pain stress on the blood-brain barrier permeability in rats

Effect of surgical pain stress on the blood-brain barrier permeability was investigated in rats. The animals were divided into four groups: Group 1: control, Group 2: immobilization stress, Group 3: acute hypertension, Group 4: immobilization stress + surgical pain stress.Bilateral hid paw surgical wounds for cannulations were applied in animals' inguinal regions under diethyl-ether anesthesia, then the animals were awaken from anesthesia to produce surgical pain stress. Evans-blue was used as a blood-brain barrier tracer. There is no significantly blood-brain barrier breakdown after short-time immobilization stress, but after adrenalin hypertension blood-brain barrier permeability was increased especially on frontal and occipital cortices in 50% of the animals. Surgical pain stress increased blood-brain barrier permeabiliy in comparison to acute adrenalin-induced hypertension (p < 0.01). In surgical pain stress-induced animals distinct Evans-blue leakage was observed in the occipital, frontal and parieto-temporal cortices.     

Oven,Temperature-Controlled Microwave,and Shade Drying Effects on Drying Kinetics,Bioactive Compounds and Antioxidant Activity of Knotweed (Polygonum cognatum Meissn.)

In this study, the plant node was dried in an oven (40, 50 and 60 °C), shade and temperature-controlled microwave (40, 50 and 60 °C) methods. Statistically (p<0.05), the values closest to the color values of fresh grass were determined in an oven at 40 °C drying temperature. Effective diffusion values varied between 8.85×10⁻⁸–5.65×10⁻⁶ m² s⁻¹. While the activation energy was 61.28 kJ mol⁻¹ in the oven, it was calculated as 85.24 kJ mol⁻¹ in the temperature-controlled microwave. Drying data was best estimated in the Midilli-Küçük (R² 0.9998) model oven at 50 °C. The highest SMER value was calculated as 0.0098 kg kWh⁻¹ in the temperature-controlled microwave drying method. The lowest SEC value in the temperature-controlled microwave was determined as 24.03 kWh kg⁻¹. It was determined that enthalpy values varied between −2484.66/−2623.38 kJ mol⁻¹, entropy values between −162.04/−122.65 J mol⁻¹ and Gibbs free energy values between 453335.22–362581.40 kJ mol⁻¹. Drying rate values were calculated in the range of 0.0127–0.9820 g moisture g dry matter⁻¹ in the temperature-controlled microwave, 0.0003–0.0762 g dry matter⁻¹ in the oven, and 0.001–0.0058 g moisture g dry moisture matter⁻¹ in the shade. Phenolic content 6957.79 μg GAE g⁻¹ fw - 48322.27 μg GAE g⁻¹ dw, flavonoid content 3806.67 mg KE L⁻¹ fw - 22200.00 mg KE L⁻¹ dw and antioxidant capacity 43.35 μmol TE g⁻¹ fw - 323.47 μmol TE g⁻¹ dw. The highest chlorophyll values were obtained from samples dried in an oven at 40 °C. According to the findings, it is recommended to dry the knotweed (Polygonum cognatum Meissn.) plant in a temperature-controlled microwave oven at low temperatures. In this study, in terms of drying kinetics and energy parameters, a temperature-controlled microwave dryer of 60 °C is recommended, while in terms of quality characteristics, oven 40 °C and shade methods are recommended.     

Efficient transfection of mouse-derived C2C12 myoblasts using a matrigel basement membrane matrix

Myogenic cell lines have been used widely in the study of myogenic differentiation, muscle regeneration and homeostasis, but, myoblasts and myotubes are difficult to transfect using conventional techniques. We have used liposome-based transfection method to introduce a green fluorescence protein (GFP)-expressing plasmid into Matrigel basement membrane matrix-coated C2C12 mouse myoblast cells. Myoblasts adhered and proliferated more rapidly on a Matrigel; thus, a dramatic increase in transfection efficiency can be obtained compared to Matrigel-untreated cells. Transfection efficiency was determined by counting fluorescent and total cells from six random fields for each condition. This protocol results in efficient (up to 60–70%) transfection of C2C12 myoblasts, high levels of GFP expression and low rate of cell death (10%). This technique is rapid, reliable, uses a lipid-based transfection reagent, and yields high transfection rates in a previously hard-to-transfect cell type.     

Dynamic Fluctuations Provide the Basis of a Conformational Switch Mechanism in Apo Cyclic AMP Receptor Protein

Escherichia coli cyclic AMP Receptor Protein (CRP) undergoes conformational changes with cAMP binding and allosterically promotes CRP to bind specifically to the DNA. In that, the structural and dynamic properties of apo CRP prior to cAMP binding are of interest for the comprehension of the activation mechanism. Here, the dynamics of apo CRP monomer/dimer and holo CRP dimer were studied by Molecular Dynamics (MD) simulations and Gaussian Network Model (GNM). The interplay of the inter-domain hinge with the cAMP and DNA binding domains are pre-disposed in the apo state as a conformational switch in the CRP''s allosteric communication mechanism. The hinge at L134-D138 displaying intra- and inter-subunit coupled fluctuations with the cAMP and DNA binding domains leads to the emergence of stronger coupled fluctuations between the two domains and describes an on state. The flexible regions at K52-E58, P154/D155 and I175 maintain the dynamic coupling of the two domains. With a shift in the inter-domain hinge position towards the N terminus, nevertheless, the latter correlations between the domains loosen and become disordered; L134-D138 dynamically interacts only with the cAMP and DNA binding domains of its own subunit, and an off state is assumed. We present a mechanistic view on how the structural dynamic units are hierarchically built for the allosteric functional mechanism; from apo CRP monomer to apo-to-holo CRP dimers.     

A scalable pipeline for highly effective genetic modification of a malaria parasite

In malaria parasites, the systematic experimental validation of drug and vaccine targets by reverse genetics is constrained by the inefficiency of homologous recombination and by the difficulty of manipulating adenine and thymine (A+T)-rich DNA of most Plasmodium species in Escherichia coli. We overcame these roadblocks by creating a high-integrity library of Plasmodium berghei genomic DNA (>77% A+T content) in a bacteriophage N15-based vector that can be modified efficiently using the lambda Red method of recombineering. We built a pipeline for generating P. berghei genetic modification vectors at genome scale in serial liquid cultures on 96-well plates. Vectors have long homology arms, which increase recombination frequency up to tenfold over conventional designs. The feasibility of efficient genetic modification at scale will stimulate collaborative, genome-wide knockout and tagging programs for P. berghei.