Recovery from ethyl methanesulphonate-induced genetical damage in barley. Independence of sodium azide treatment

Barley seeds were treated for 3 h at 25°C with 240 mM ethyl methanesulphonate (EMS), washed for 18 h, treated with various concentrations of unbuffered sodium azide (pH 6.7–7.3) for 3 h at 25°C, re-dried to 30% water content and either sown immediately or stored at 25°C for 12 days and then sown. The synergistic action of sodium azide post-treatment has been demonstrated only for the EMS-induced M₁ germination reduction, while the EMS-induced M₁ sterility and the yield of M₂ chlorophyll mutants were unaffected. The ?storage” recovery from EMS-induced mutagenic effects was insensitive to sodium azide post-treatment. The 12 day-seed storage at 25°C brought about an improvement of M₁ germination, M₁ survival, M₁ fertility and a decrease in the amount of M₂ mutants, regardless of whether sodium azide post-treatment was applied or not.     

A contribution to the phenotype distribution of phosphoglucomutase in Czechoslovakia (the district of České Budějovice)

Summary A population sample of 416 unrelated donors from eské Budjovice (southorn Bohemia) was investigated for the phenotypes of phosphoglucomutase (PGM). The calculated frequencies of the allelles PGM ₁ ¹ and PGM ₁ ² , 0.770 and 0.230, respectively, correspond to the expected frequencies of the phenotypes PGM 1=0.593, PGM 2-1=0.354, and PGM 2=0.0529. No rare phonotype was detected.     

Studien über dieJungermannioideae (Hepaticae) 8.Jungermannia subg.Plectocolea und subg.Solenostoma in Australien,Neuseeland und Ozeanien

Die achte Studie über die SubfamilieJungermannioideae (Jungermanniaceae, Hepaticae) ist dén Arten der GattungJungermannia L. emend.Dum., die in Australien, Neuseeland und Ozeanien vorkommen, gewidmet. Die meist endemischen Arten, die auf Hawaii vorkommen, werden als eine selbständige (neunte) Studie bearbeitet. Im genannten Gebiet wurden 13 Arten der GattungJungermannia L. emend.Dum. festgestellt.J. (P.) hasskarliana (Nees) Steph.,J. (P.) obliquifolia (Schiffn.) Váňa,J. (P.) tetragona Lindenb.,J. (P.) hirticalyx Steph.,J. (P.) minutiverrucosa Amak.,J. (P.) wattsiana Steph.,J. (S.) ariadne Tayl.,J. (S.) totipapillosa Hodgs.,J. (S.) inundata Hook. fil. etTayl. emend.Mitt. undJ. (S.) orbiculata (Col.) Grolle sind eingehend taxonomisch bearbeitet,J. (P.) micrantha (Mitt.) Steph. wird in die nächste Studie eingereiht.J. (P.) boninensis (Horik.) Inoue wurde vonInoue (Inoue etIwatsuki 1969) eingehend beschrieben und abgebildet,Haplozia comptonii Pears. wurde schon im 2. Teil diskutiert. Die anderen aus dem Gebiet bekannten Arten sind mit den obenerwähnten Arten synonymisiert oder zu anderen Gattungen gestellt.     

Oxidative stress-resistance assay for screening yeast strains overproducing heterologous proteins

Many natural proteins have been developed into drugs and produced for direct application. Identifying improved hosts to achieve high-level heterologous protein production is a challenge in the study of heterologous protein expression in recombinant yeast. In this study, a novel high-throughput assay to screen such overproducing Saccharomyces cerevisiae strains was systematically developed. The protocol designed was based on screening host strain derivatives with increased superoxide dismutase dependent resistance to oxidative stress. Yeast cells transformed with recombinant plasmid carrying SOD1 gene as a reporter responded exquisitely to oxidative stress induced by elevated concentrations of paraquat. Improved yeast strains resulting from screening clones subjected to genome shuffling through selective pressure argue for a more effective screening system compared with traditonal selection. Moreover, this approach can be employed in general biochemical analysis without utilization of flow cytometry or well plate reader. Therefore, it is expected that the high-throughput assay would make superior strains producing heterologous proteins.     

A simple process for the isolation of epithelial cells from bacteria-contaminated samples using anchoring molecules

A simple and efficient tool to isolate epithelial cells from bacteria-contaminated samples has been developed using two different microparticles functionalized with chemical molecules. The epithelial cells could be captured simply by biocompatible anchors for membranes (BAM), consisting of poly(ethylene glycol) functionalized with oleyl-chain-conjugated NHS (N-hydroxysuccinimide) on glass microparticles, whereas bacteria were adsorbed on 3-aminopropyltrimethoxysilane (ATPS)-functionalized magnetic microparticles. In the case of samples highly contaminated with bacteria, epithelial cells were not isolated successfully by both of the single BAM- and antibody-functionalized microparticles. Therefore, serial isolation steps of these two different chemical functionalized microparticles were introduced. The concentration of bacteria was decreased dramatically by using APTS-functionalized magnetic particles prior to the isolation of epithelial cells by BAM microparticles. With these serial processes, successful isolation of epithelial cells was achieved from bacteria-contaminated epithelial samples. The applicability of this method was verified with bacteria-contaminated intestinal samples biopsied from a BALB/C mouse for primary cell cultivation.     

A Comprehensive Analysis of Symmetric Arginine Dimethylation in Colorectal Cancer Tissues Using Immunoaffinity Enrichment and Mass Spectrometry

Protein arginine methyltransferase 5 (PRMT5) is a major enzyme responsible for generating monomethyl and symmetric dimethyl arginine in proteins. PRMT5 is essential for cell viability and development, and its overexpression is observed in a variety of cancers. In the present study, it is found that levels of PRMT5 protein and symmetric arginine dimethylation in colorectal cancer (CRC) tissues are increased compared to those in adjacent noncancerous tissues. Using immunoaffinity enrichment of methylated peptides combined with high‐resolution mass spectrometry, a total of 147 symmetric dimethyl‐arginine (SDMA) sites in 94 proteins are identified, many of which are RNA binding proteins and enzymes. Quantitative analysis comparing CRC and normal tissues reveals significant increase in the symmetric dimethylation of 70 arginine sites in 46 proteins and a decrease in that of four arginine sites in four proteins. Among the 94 proteins identified in this study, it is confirmed that KH‐type splicing regulatory protein is a target of PRMT5 and highly expressed in CRC tissues compared to noncancerous tissues. This study is the first comprehensive analysis of symmetric arginine dimethylation using clinical samples and extends the number of known in vivo SDMA sites. The data obtained are available via ProteomeXchange with the identifier PXD015653.