Are seeds suitable for flow cytometric estimation of plant genome size?

BACKGROUND: Nuclear DNA content in plants is commonly estimated using flow cytometry (FCM). Plant material suitable for FCM measurement should contain the majority of its cells arrested in the G0/G1 phase of the cell cycle. Usually young, rapidly growing leaves are used for analysis. However, in some cases seeds would be more convenient because they can be easily transported and analyzed without the delays and additional costs required to raise seedlings. Using seeds would be particularly suitable for species that contain leaf cytosol compounds affecting fluorochrome accessibility to the DNA. Therefore, the usefulness of seeds or their specific tissues for FCM genome size estimation was investigated, and the results are presented here. METHODS: The genome size of six plant species was determined by FCM using intercalating fluorochrome propidium iodide for staining isolated nuclei. Young leaves and different seed tissues were used as experimental material. Pisum sativum cv. Set (2C = 9.11 pg) was used as an internal standard. For isolation of nuclei from species containing compounds that interfere with propidium iodide intercalation and/or fluorescence, buffers were used supplemented with reductants. RESULTS: For Anethum graveolens, Beta vulgaris, and Zea mays, cytometrically estimated genome size was the same in seeds and leaves. For Helianthus annuus, different values for DNA amounts in seeds and in leaves were obtained when using all but one of four nuclei isolation buffers. For Brassica napus var. oleifera, none of the applied nuclei isolation buffers eliminated differences in genome size determined in the seeds and leaves. CONCLUSIONS: The genome size of species that do not contain compounds that influence fluorochrome accessibility appears to be the same when estimated using specific seed tissues and young leaves. Seeds can be more suitable than leaves, especially for species containing staining inhibitors in the leaf cytosol. Thus, use of seeds for FCM nuclear DNA content estimation is recommended, although for some species a specific seed tissue (usually the radicle) should be used. Protocols for preparation of samples from endospermic and endospermless seeds have been developed.     

137Cs and 40K isotopes in forest and wasteland soils in a selected region of eastern Poland 20 years after the Chernobyl accident

The vertical ¹³⁷Cs profile of forest and wasteland soils was analyzed in the south of the Podlasie Lowland area (Eastern Poland) about 20 years after the Chernobyl accident. In addition, the concentration of ⁴⁰K in soils of the investigated area was measured. Below the litter layer (mean thickness 3 cm), the soil samples were collected up to a depth of 12 cm and then divided into three layers: 0–3, 3–7, 7–12 cm. The behavior of ¹³⁷Cs and ⁴⁰K isotopes in soils was analyzed depending on the depth from which the soil samples were collected, as well as on the content of organic carbon, pH of soil and its granulometric composition. It was established that the density of ¹³⁷Cs in the litter layer equals 2.17 kBq m⁻²; it is the highest in layer 0–3 cm where it equals 3.44 kBq m⁻², and it decreases with the depth to the value of 0.76 kBq m⁻² in layer 7–12 cm. No similar pattern was observed in wasteland soils. The concentrations of ⁴⁰K in forest and wasteland soils did not change significantly with depth.     

Anemia secondary to valproic acid therapy in a 13- year-old boy: a case report

ABSTRACT: INTRODUCTION: Valproic acid is a commonly used anti-epileptic drug. Hematological toxicities are among theoccasionally observed adverse effects of this medication. CASE PRESENTATION: We present the case of a 13-year-old Caucasian boy who demonstrated mild anemia 12months after the introduction of valproic acid therapy. A bone marrow biopsy revealedmaturation arrest of proerythroblasts. CONCLUSION: Prompt diagnosis and valproic acid discontinuation resulted in the patient's recovery.     

Phytoplankton composition and cell carbon distribution in Prydz Bay, Antarctica: relation to organic particulate matter and its {delta}13C values

In January-February 1991, in Prydz Bay, phytoplankton bloomwas evident in the inner shelf area with the dominant diatomsbeing represented mainly by pennate species of the Nitzschia-Fragilariopsisgroup. Dinoflagellates and naked flagellates were most abundantin the centre of the bay; however, larger heterotrophic speciesprevailed at the southern stations. Cell carbon values (average317 µg l^–1; range 92-1048 µg l^–1) foundin the bloom in the south were chiefly due to pennate diatomsand larger heterotrophic dinoflagellates. Much lower carbonvalues (average 51 µg l^–1; range 7-147 µgl^–1) in the outer shelf region were mainly contributedby large centric diatoms (70-110 mu;m) and small dinoflagellates(5-25 µm). Wide ranges of algal cell sizes were observedin both southern and northern communities; the overlapping ofsizes of diatoms and flagellates, the latter containing heterotrophs,suggested complex trophic relationships within the planktonand an enhanced heterotrophic activity in the south. North-to-southvariations in surface     

The organellar peptidasome,PreP: A journey from Arabidopsis to Alzheimer's disease

The novel peptidasome, called presequence protease, PreP, was originally identified and characterized in Arabidopsis thaliana as a mitochondrial matrix and chloroplast stroma localized metalloprotease. PreP has a function as the organellar peptide clearing protease and is responsible for degrading free targeting peptides and also other unstructured peptides up to 65 amino acid residues that might be toxic to organellar functions. PreP contains an inverted Zn-binding motif and belongs to the pitrilysin protease family. The crystal structure of AtPreP refined at 2.1 Å demonstrated a unique totally enclosed large cavity of 10 000 Å³ that opens and closes in response to peptide binding, revealing a novel catalytic mechanism for proteolysis. Homologues of PreP have been found in yeast and human mitochondria. Interestingly, the human PreP, hPreP, is the protease that is responsible for clearing the human brain mitochondria from the toxic amyloid-β peptide (Aβ) associated with Alzheimer's disease (AD). Accumulation of Aβ has been shown in the brain mitochondria from AD patients and mutant transgenic mice overexpressing Aβ. Here, we present a review of our present knowledge on structural and functional characteristics of PreP and discuss its mitochondrial Aβ-degrading activity in the human brain mitochondria in relation to AD.     

Inside Cover: Localization of PD‐L1 on single cancer cells by iSERS microscopy with Au/Au core/satellite nanoparticles (J. Biophotonics 3/2020)

iSERS (SERS=surface‐enhanced Raman scattering) microscopy is an emerging Raman‐based staining technique for the selective localization of target proteins on cells and tissues using antibody‐ SERS nanotag conjugates. In this contribution we demonstrate the feasibility of iSERS for imaging of programmed cell death‐ligand 1 (PD‐L1), an important predictive biomarker, on single SkBr‐3 breast cancer cells. Further details can be found in the article by Elzbieta Stepula, Matthias König, Xin‐Ping Wang, et al. ( e201960034 ).

     

Polypeptides from maize seedlings with protein kinase functions

Low molecular weight protein kinase from maize seedlings was isolated. The molecular weight of the enzyme was below 10 000. The enzyme preferentially utilized ATP and casein as donor and acceptor of phosphorus, respectively. Casein kinase was a cyclic nucleotide-independent, heparin-sensitive enzyme. The low molecular weight casein kinase was resolved into basic, neutral and slightly acidic peaks of activity by chromatofocussing.     

Serum alkaline phosphatase polymorphism in pigs1

Genetic variants of serum alkaline phosphatase were studied by the method of starch gel electrophoresis in the Zlotnicka Pstra breed of pigs. Two regions of alkaline phosphatase migration were observed. A single fraction in region I and four different phenotypes: AB, B, BC and BD in region II, were found. For AB, B and BC phenotypes the genetic control by three alleles AkpA, AkpB and AkpC in suggested. The observed segregation ratios in some cases deviated significantly from the expected ones.