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Authors: M. Duleba and Zs. Trábert
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A new lichenicolous fungus Buelliella indica colonising on the thallus of Graphis longiramea is described from the state of Nagaland, a part of the Indo-Burma biodiversity hotspot region in India. It is characterised by its brown epihymenium, much smaller ascospores with dimensions of 11.5–13.8 × 4.8–6 µm and the new host Graphis.

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In the present study, morphological and anatomical structures of cypsela – 12 Cirsium Miller (Carduoideae, Asteraceae) taxa belonging to two sections (sect. Cirsium and sect. Cephalonoplos) were investigated in detail with using stereomicroscope and light microscope. The taxa were evaluated comparatively in the aspect of carpological variations and their anatomies were presented in here for the first time. Morphological features including size, shape and colour of cypselae were examined. From anatomical observations, anatomical structures of pericarp, as well as the structure of testa were described. Cypselae colours differ from light brown to stramineous, sometimes with blackish striations. Their shapes change from oblong to oblanceolate, rarely obovate. The largest cypselae are present in C. echinus (1.59±0.03 mm × 4.68±0.07 mm) and the smallest ones are found in C. subinerme (1.20±0.02 mm × 2.97±0.05 mm). The pericarp is characterised by almost parenchymatous cells, while the testa is composed of lignified sclerenchymatous cell lines and crushed cells group. Secretory structure in testa bundle was evaluated. Results obtained from this study were compared with the present data in literature. Overall, morphological and anatomical characteristics of cypselae provide useful taxonomic markers in their classifications of the studied taxa of Cirsium but not distinctive for their sectional levels.

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Background and aims

Targeted chemotherapeutics such as cetuximab can cause many side effects such as skin toxicity when used in high concentrations. In addition, cancer cells can develop resistance to some of the anticancer agents during treatment. The lack of the desired success in chemotherapy and the development of resistance to chemotherapeutics, such as epirubicin HCl, suggest that there is a need for combined therapies. The combination of targeted chemotherapeutics and conventional chemotherapy drugs may lead to the emergence of new strategies in the treatment of cancer. In this study, cytotoxic, antiproliferative, cell cycle inhibitive, oxidative stress generation, and apoptotic effects and effect mechanisms of cetuximab alone and together with epirubicin HCl on parental liver cancer cells (P-Hep G2) and epirubicin HCl-resistant liver cancer cells (R-Hep G2) were investigated.

Materials

Cytotoxic effects of cetuximab alone and with epirubicin-HCl on cells were determined by Cell Titer-Blue® Cell Viability and Lactate Dehydrogenase Activity tests. Cell cycle distributions and apoptosis were detected by reverse transcription polymerase chain reaction (RT-PCR).

Results

Cetuximab with epirubicin HCl treatment increased the cytotoxic effect on both cells. Caspase-3/7 activity increased 3 and 1.5 times in comparison with control group in P-Hep G2 and R-Hep G2 cells, respectively, after treating with cetuximab alone, whereas the increase was found to be approximately 4.7 and 2.5 times when cetuximab was treated with epirubicin HCl in P-Hep G2 and R-Hep G2 cells, respectively. Both cetuximab alone and together with epirubicin HCl treatments caused increases in Bax/Bcl-2 ratio in both cells.

Discussion

Treatment of cetuximab with epirubicin HCl to P-Hep G2 and R-Hep G2 cells was found to be more effective in cytotoxic effect and inducing apoptosis comparison to cetuximab alone treatment. In addition, combination treatment showed different effects on pro-apoptotic/anti-apoptotic genes expression according to cells drug resistance properties.

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Powdery mildew (Blumeria graminis f. sp. tritici) is one of the most devastating wheat diseases. The wheat line N9134 contains PmAS846 that was transferred to N9134 from wild emmer wheat, and is still one of the most effective resistance genes in China. A full-length wheat RPM1 gene was obtained by rapid amplification of cDNA ends (RACE) based on the up-regulated probe sequence from differentially expressed transcripts during the N9134 and powdery mildew interaction. The gene was named TaRPM1, and the open reading frame (ORF) is 2721 nucleotides and encodes a polypeptide of 907 amino acids with a predicted isoelectric point of 4.86. Phylogenetic analysis revealed that TaRPM1 was highly homologous on both Aegilops tauschii and Triticum urartu at both the nucleotide and protein level. Using real-time quantitative PCR, the TaRPM1 gene expression level in wheat leaves was found to be sharply up-regulated, while the transcript level was lowly induced in the root and stem. Under the powdery mildew treatment, the transcription profile of TaRPM1 was very strongly expressed at 48 hour post inoculation (hpi), which increased again to 96 hpi and reaching a high level at 120 hpi. Based on sequence similarities and positions, we inferred that the TaRPM1 gene was on wheat chromosome 3D. These results suggested that TaRPM1 plays an important role in the mechanism of innate immunity to infection by the powdery mildew pathogen.

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This study aimed at evaluating in outpatients an algorithm for the laboratory diagnosis of Clostridioides (Clostridium) difficile infection (CDI), i.e., enzyme immunoassays (EIAs) detecting bacterial glutamate dehydrogenase (GDH) and toxin A/B, followed by polymerase chain reaction (PCR) analyses of samples with discordant EIA results.

In total, 9802 examinations of stool samples by GDH and toxin EIAs performed in 7263 outpatients and 488 inpatients were analyzed retrospectively. Samples with discordant EIA results had been tested by a commercially available PCR assay detecting genes of the C. difficile-specific triose phosphate isomerase (tpi) and toxin B (tcdB).

Concordant EIA results (686 C. difficile-positive, 8121 negative) were observed for 8807 (89.8%; 95% CI, 89.2–90.4%) samples. Of 958 samples with discordant EIA results, 895 were analyzed using PCR and 580 of 854 GDH-positive/borderline, toxin-negative samples (67.9%; 95% CI, 64.7–71.0%) were positive for tpi and tcdB, while 274 samples (32.1%; 95% CI, 29.0–35.3%) were tcdB-negative. In contrast, 35 of 41 GDH-negative, toxin-positive/borderline samples (85.4%; 95% CI, 71.2–93.5%) were tcdB-negative. Still, 6 samples (14.6%; 95% CI, 6.5–28.8%) yielded positive PCR results for both genes.

In conclusion, around 90% of the samples were analyzed appropriately by only applying EIAs. Approximately one third of the PCR-analyzed samples were tcdB-negative; thus, patients most likely did not require CDI treatment.

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Authors: J. Sangrattanaprasert, S. Kornochaleart and S. Chantanaorrapint

Cololejeunea metzgeriopsis (K. I. Goebel) Gradst. et al., a neotenic liverwort, was newly discovered in lowland evergreen forest, southern Thailand. A description and illustrations of Thai plants are provided.

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Authors: S. Esfandani-Bozchaloyi, M. Sheidai and M. Hassanzadeh Kalalegh

The genus Geranium (Geraniaceae); with about 320 species throughout the temperate regions, is chemically characterised by the presence of tannins, flavonoids, anthocyanins and essential oils which interfere with the extraction of pure genomic DNA. It is necessary to optimise the extraction protocols to reduce the effects of the presence of these compounds to the lowest level.

The present study compares the plant genomic DNA extraction Kit (DNP™ Kit), CTAB DNA extraction method by Murray and Thompson and Sahu et al., from the extracting DNA point of view Geranium species. The results showed significant differences in DNA contents between the three methods. Quantity and quality of extracted genomic DNAs were compared by employing the spectrophotometer, Nano-Drop, agarose gel electrophoresis, and polymerase chain reaction (PCR) methods and molecular marker such as (ITS and trnL-F) and ISSR. The method of Sahu et al., provided the best results (200 ng/µL) in terms of quantity and quality of DNA, therefore, this method was taken and optimised for DNA extraction. Our results proposed that this method could be effective for plants with same polysaccharides, proteins and polyphenols components. The advantage of this method is that it omits the use of liquid nitrogen and toxic phenols which are expensive. The success of this method in obtaining high-quality genomic DNA has been demonstrated in the Geranium species group and the reliability of this method has been discussed.

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Powdery mildew, caused by Blumeria graminis f. sp. hordei (Bgh) is a common foliar disease of barley worldwide. The creation of new cultivars with durable resistance to Bgh is highly desirable. This work was undertaken to examine the resistance to Bgh in 10 genetically diverse barley parents, and to evaluate their general combining ability (GCA) and specific combining ability (SCA) effects toward determining the genetic basis of disease resistance. Two experiments, in a growth chamber on seedling and in the field on adult plant stages, were conducted using a randomized complete block design with three replicates. The parents expressing differences in their reactions to Bgh were crossed in a half-diallel mating design to generate 45 full-sib families. Genetic component analysis showed significant effects for both GCA and SCA under both experiments suggesting that additive as well as non-additive genetic mechanisms were involved in the expression of resistance in these parents. The estimate of narrow-sense heritability was 0.63 and broad-sense heritability was 98% indicating that selection for the disease resistance should be effective in these crosses. Resistant parents ‘Banteng, PK 30-136 and ‘Igri’ had significantly negative GCA effects, suggesting their prime suitability for use in barley breeding programs to improve resistance to Bgh.

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Glyphosate (N-(phosphonomethyl)glycine) is the most-used herbicide worldwide. Many studies in the past have shown that residues of the herbicide can be found in many cultivated plants, including those used as livestock feed. Sensitivity to glyphosate varies with bacteria, particularly those residing in the intestine, where microbiota is exposed to glyphosate residues. Therefore, less susceptible pathogenic isolates could have a distinct advantage compared to more sensitive commensal isolates, probably leading to dysbiosis.

To determine whether the ruminal growth and survival of pathogenic Escherichia coli or Salmonella serovar Typhimurium are higher when glyphosate residues are present in the feed, an in vitro fermentation trial with a “Rumen Simulation System” (RUSITEC) and a glyphosate-containing commercial formulation was performed.

Colony forming units of E. coli and Salmonella ser. Typhimurium decreased steadily in all fermenters, regardless of the herbicide application. Minimum inhibitory concentrations of the studied Salmonella and E. coli strains did not change, and antibiotic susceptibility varied only slightly but independent of the glyphosate application.

Overall, application of the glyphosate-containing formulation in a worst-case concentration of 10 mg/L neither increased the abundance for the tested E. coli and Salmonella strain in the in vitro fermentation system, nor promoted resistance to glyphosate or antibiotics.

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