UMP kinase activity is involved in proper chloroplast development in rice

Isolation of leaf-color mutants is important in understanding the mechanisms of chloroplast biogenesis and development. In this study, we identified and characterized a rice (Oryza sativa) mutant, yellow leaf 2 (yl2), exhibiting pale yellow leaves with a few longitudinal white stripes at the early seedling stage then gradually turning yellow. Genetic analyses revealed that YL2 encodes a thylakoid membrane-localized protein with significant sequence similarity to UMP kinase proteins in prokaryotes and eukaryotes. Prokaryotic UMP kinase activity was subsequently confirmed, with YL2 deficiency causing a significant reduction in chlorophyll accumulation and photochemical efficiency. Moreover, YL2 is also light dependent and preferentially expressed in green tissues. Chloroplast development was abnormal in the yl2 mutant, possibly due to reduced accumulation of thylakoid membranes and a lack of normal stroma lamellae. 2D Blue-Native SDS-PAGE and immunoblot analyses revealed a reduction in several subunits of photosynthetic complexes, in particular, the AtpB subunit of ATP synthase, while mRNA levels of corresponding genes were unchanged or increased compared with the wild type. In addition, we observed a significant decrease (ca. 36.3%) in cpATPase activity in the yl2 mutant compared with the wild type. Taken together, our results suggest that UMP kinase activity plays an essential role in chloroplast development and regulating cpATPase biogenesis in rice.     

Enhancement of thiamine release during synthetic mutualism between <Emphasis Type="Italic">Chlorella sorokiniana</Emphasis> and <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> growing under stress conditions

Thiamine release during synthetic mutualism between Chlorella sorokiniana co-immobilized in alginate beads with the microalgae growth-promoting bacterium Azospirillum brasilense was measured under stress conditions of pH, light intensity, and nitrogen starvation in short-term experiments. Thiamine release in the co-immobilized treatment was significantly higher at acidic pH compared to thiamine released by either microorganism alone. Under slightly alkaline pH, C. sorokiniana released the highest amount of thiamine. At stressful pH 6, the co-immobilized treatment released a higher quantity of thiamine than the sum of thiamine released by either microorganisms when immobilized separately. Release of thiamine by C. sorokiniana alone or co-immobilized was light intensity dependent; with higher the light intensity, more thiamine was released. Extreme light intensity negatively affected growth of the microalgae and release of thiamine. Nitrogen starvation during the first 24 h of culturing negatively affected release of thiamine by both microorganisms, where C. sorokiniana was more severely affected. Partial or continuous nitrogen starvation had similar negative effects on C. sorokiniana, but co-immobilization improved thiamine release. These results indicate that thiamine is released during synthetic mutualism between C. sorokiniana and A. brasilense, and this happens specifically during the alleviation of pH stress in the microalgae.     

Effects of VEGF levels on anti-VEGF therapy for patients with idiopathic choroidal neovascularization

The objective of this study is to investigate the levels of vascular endothelial growth factor (VEGF) and other cytokines in aqueous humor of patients with idiopathic choroidal neovascularization (CNV) and their effects together with central retinal thickness (CRT) on the response to intravitreal injection of anti-VEGF antibody ranibizumab. This clinical study recruited 32 eyes from 32 patients with CNV under or besides fovea. VEGF, interleukin (IL)-6, IL-8, and monocyte chemoattractant protein (MCP)-1 levels were detected in aqueous humor (0.1 ml) sampled during intravitreal injection. Aqueous humor controls were from nine cataract patients without any systemic disorders. The VEGF levels in aqueous humor were negatively related (r = ?0.373, p = 0.035) to CRT, which was positively related (r = 0.743, p < 0.001) to the number of injections. The VEGF levels before treatment and during the third injection in four patients with three or more injections were 13.42 ± 8.50 and 5.75 ± 3.68 (p = 0.055), respectively. The average best corrected visual acuity (BCVA) before and 12 months after treatment were 57.03 ± 16.15 and 75.16 ± 11.78 (p < 0.001), and the average CRT before and 12 months after treatment were 352.09 ± 84.15 and 251.13 ± 63.96 (p < 0.001), respectively. The visual improvement was negatively related (r = ?0.815, p < 0.001) to the visual baseline, and the vision 12 months after treatment was positively related (r = 0.581, p < 0.001) to that before treatment. No severe ocular or systemic complication appeared during treatment and follow-ups for all the patients. Intravitreal injection of anti-VEGF antibody ranibizumab is safe and effective for the treatment of idiopathic CNV through decreasing CRT. The patients with larger CRT baseline need more injections of ranibizumab.     

Fluorescence change of <Emphasis Type="Italic">Fusobacterium nucleatum</Emphasis> due to <Emphasis Type="Italic">Porphyromonas gingivalis</Emphasis>

The aim of this study was to measure changes in the fluorescence of Fusobacterium nucleatum interacting with Porphyromonas gingivalis for excitation with blue light at 405-nm. P. gingivalis was mono- and co-cultivated in close proximity with F. nucleatum. The fluorescence of the bacterial colonies was photographed using a QLF-D (Quantitative Light-induced Fluorescence-Digital) Biluminator camera system with a 405 nm light source and a specific filter. The red, green and blue intensities of fluorescence images were analyzed using the image analysis software. A fluorescence spectrometer was used to detect porphyrin synthesized by each bacterium. F. nucleatum, which emitted green fluorescence in single cultures, showed intense red fluorescence when it was grown in close proximity with P. gingivalis. F. nucleatum co-cultivated with P. gingivalis showed the same pattern of fluorescence peaks as for protoporphyrin IX in the red part of the spectrum. We conclude that the green fluorescence of F. nucleatum can change to red fluorescence in the presence of adjacent co-cultured with P. gingivalis, indicating that the fluorescence character of each bacterium might depend on the presence of other bacteria.     

European <Emphasis Type="Italic">Hodophilus</Emphasis> (Clavariaceae,Agaricales) species with yellow stipe

Phylogenetic reconstruction of Hodophilus species with a yellow colour on the stipe based on nrITS, nrLSU and rpb2 sequences revealed six European species. All these species correspond to the widely accepted European concept of a single species Hodophilus micaceus. Four of these species are described and illustrated. H. micaceus and H. phaeoxanthus are recognised as two separate species and H. albofloccipes as a synonym of the latter. Two species, H. anatinus and H. cambriensis, are described as new. Possible endemism of H. micaceus and H. cambriensis to the British Isles is discussed. All analysed North American samples represent different species to those found in Europe. The North American species Hygrophorus rugulosus is combined in the genus Hodophilus. The preliminary key uses position and development of the yellow colour during maturation as the most important distinguishing character. The presence of the yellow colour is discussed as a possible synapomorphic character.     

Anti-inflammatory effects of <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis> extracts on human blood mononuclear cells and murine macrophages

The microalgae Phaeodactylum tricornutum (PT) is known for its high content of omega-3 fatty acids, which are known to attenuate inflammation. Additionally, this microalga contains other nutrients such as carbohydrates, vitamins, proteins, and carotenoids and therefore could be of interest for animal and human nutrition. Here, we investigated the effects of hexane, ethanolic, and aqueous extracts on lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMCs) and on the murine macrophage cell line RAW 264.7. Further, the cytotoxicity of the extracts was studied using the MTT assay. The production of pro-inflammatory cytokines was significantly inhibited by the ethanolic and aqueous P. tricornutum extracts but not by the hexane extract. Both at the mRNA and at the protein levels, the aqueous extract inhibited LPS-induced IL-6, IL-1β, TNFα, and COX-2 expression and release by up to 96% (mRNA) and 79% (protein) in a dose-dependent manner. Compared to the aqueous extract, the ethanolic extract was less effective in cytokine inhibition. The production of nitric oxide in RAW 264.7 cells was significantly reduced by all extracts. We showed that the anti-inflammatory effect of P. tricornutum is exerted through inhibition of nuclear factor-κB activation and dependent on the mitogen-activated protein (MAP) kinase pathway. Our data indicate anti-inflammatory effects of the aqueous P. tricornutum extract and provide a basis information on the safety and potential health benefits of P. tricornutum usage for future animal and human nutrition.     

Development and Characterization of Nisin Nanoparticles as Potential Alternative for the Recurrent Vaginal Candidiasis Treatment

The aim of this work was the development and characterization of nisin-loaded nanoparticles and the evaluation of its potential antifungal activity. Candidiasis is a fungal infection caused by Candida sp. considered as one of the major public health problem currently. The discovery of antifungal agents that present a reduced or null resistance of Candida sp. and the development of more efficient drug release mechanisms are necessary for the improvement of candidiasis treatment. Nisin, a bacteriocin commercially available for more than 50 years, exhibits antibacterial action in food products with potential antifungal activity. Among several alternatives used to modulate antifungal activity of bacteriocins, polymeric nanoparticles have received great attention due to an effective drug release control and reduction of therapeutic dose, besides the minimization of adverse effects by the preferential accumulation in specific tissues. The nisin nanoparticles were prepared by double emulsification and solvent evaporation methods. Nanoparticles were characterized by dynamic light scattering, zeta potential, Fourier transform infrared, X-ray diffraction, differential scanning calorimetry, and scanning electron microscopy. Antifungal activity was accessed by pour plate method and cell counting using Candida albicans strains. The in vitro release profile and in vitro permeation studies were performed using dialysis bag method and pig vaginal mucosa in Franz diffusion cell, respectively. The results revealed nisin nanoparticles (300 nm) with spherical shape and high loading efficiency (93.88?±?3.26%). In vitro test results suggest a promising application of these nanosystems as a prophylactic agent in recurrent vulvovaginal candidiasis and other gynecological diseases.     

<Emphasis Type="Italic">Entransia</Emphasis> and <Emphasis Type="Italic">Hormidiella</Emphasis>, sister lineages of <Emphasis Type="Italic">Klebsormidium</Emphasis> (Streptophyta), respond differently to light,temperature, and desiccation stress

The green-algal class Klebsormidiophyceae (Streptophyta), which occurs worldwide, comprises the genera Klebsormidium, Interfilum, Entransia, and Hormidiella. Ecophysiological research has so far focused on the first two genera because they are abundant in biological soil crust communities. The present study investigated the photosynthetic performances of Hormidiella attenuata and two strains of Entransia fimbriata under light, temperature, and desiccation stress. Their ultrastructure was compared using transmission electron microscopy. The two Entransia strains showed similar physiological responses. They used light more efficiently than Hormidiella, as indicated by higher oxygen production and relative electron transport rate under low light conditions, lower light saturation and compensation points, and higher maximum oxygen production during light saturation. Their requirement for low light levels explains the restriction of Entransia to dim limnetic habitats. In contrast, Hormidiella, which prefers drier soil habitats, responded to light gradients similarly to other aero-terrestrial green algae. Compared to Entransia, Hormidiella was less affected by short-term desiccation, and rehydration allowed full recovery of the photosynthetic performance. Nevertheless, both strains of Entransia coped with low water availability better than other freshwater algae. Photosynthetic oxygen production in relation to respiratory consumption was higher in low temperatures (Entransia: 5 °C, Hormidiella: 10 °C) and the ratio decreased with increasing temperatures. Hormidiella exhibited conspicuous triangular spaces in the cell wall corners, which were filled either with undulating cell wall material or with various inclusions. These structures are commonly seen in various members of Klebsormidiophyceae. The data revealed significant differences between Hormidiella and Entransia, but appropriate adaptations to their respective habitats.     

Four new species of <Emphasis Type="Italic">Trichoderma</Emphasis> with hyaline ascospores from central China

Collections of Trichoderma producing hyaline ascospores from central China were examined. Four new species, Trichoderma asterineum, T. henanense, T. odoratum and T. pseudobritdaniae, were discovered, described and illustrated. Their phylogenetic positions were explored based on sequence analyses of the combined RNA polymerase II subunit b (rpb2) and translation elongation factor 1 alpha (tef1) genes. As a sister of T. leguminosarum, T. asterineum can be easily recognised by its pale yellow stromata, ochre to brown ostiolar dots surrounded by stellate cracks, green conidia and slow growth. Trichoderma henanense is distinctive in pulvinate or discoid, dirty yellow to brownish yellow stromata, brown to dark brown ostiolar dots, small monomorphic ascospores in relatively short asci and white colonies with dense aerial hyphae in cultures. Trichoderma odoratum forms an independent lineage as a sister of T. henanense and is characterised by yellow to greyish yellow, pulvinate stromata with dark brown or reddish brown projecting ostiolar dots, slow growth, trichoderma- to verticillium-like conidiophores, hyaline conidia and producing a mushroom-like odour in culture. Trichoderma pseudobritdaniae is closely associated with but easily separated from T. britdaniae in pulvinate, brownish yellow or greyish yellow stromata with dark brown or grey black ostiolar dots, relatively large perithecia, monomorphic ascospores, somewhat low growth rate, trichoderma- to verticillium-like conidiophores and hyaline conidia. Morphological distinctions and sequence divergences between the new species and their close relatives are discussed.     

Effect of light quality on the growth and proximal composition of <Emphasis Type="Italic">Amphora</Emphasis> sp.

We determined the effects of various light spectra (white, blue, green, yellow, and Grolux) on the growth rate, proximate composition, pigment content, and cell size of the marine benthic diatom Amphora sp. during two growth phases of batch cultures. The growth rate was higher under green light and lowest with Grolux and yellow light. Cell size differed significantly between growth phases but remained unaffected by light spectra; the smallest cells were observed on the initial day of culture, whereas larger cells developed in the stationary phase under Grolux treatment. The proximate composition was modified by the light spectra and growth phase. In the exponential growth phase, the protein content was higher with yellow and white light, and lipid content peaked with Grolux. The pigment content (chlorophyll a, carotenoids) was also higher under yellow light. In the stationary growth phase, we noted a higher carbohydrate content under Grolux and yellow light. Our results show that light spectra and growth phase influence the metabolism of Amphora sp., changing its proximate composition, pigment content, growth rate, and cell size.     

Estuarine fouling communities are dominated by nonindigenous species in the presence of an invasive crab

Interactions between anthropogenic disturbances and introduced and native species can shift ecological communities, potentially leading to the successful establishment of additional invaders. Since its discovery in New Jersey in 1988, the Asian shore crab (Hemigrapsus sanguineus) has continued to expand its range, invading estuarine and coastal habitats in eastern North America. In estuarine environments, H. sanguineus occupies similar habitats to native, panopeid mud crabs. These crabs, and a variety of fouling organisms (both NIS and native), often inhabit man-made substrates (like piers and riprap) and anthropogenic debris. In a series of in situ experiments at a closed dock in southwestern Long Island (New York, USA), we documented the impacts of these native and introduced crabs on hard-substrate fouling communities. We found that while the presence of native mud crabs did not significantly influence the succession of fouling communities compared to caged and uncaged controls, the presence of introduced H. sanguineus reduced the biomass of native tunicates (particularly Molgula manhattensis), relative to caged controls. Moreover, the presence of H. sanguineus favored fouling communities dominated by introduced tunicates (especially Botrylloides violaceous and Diplosoma listerianum). Altogether, our results suggest that H. sanguineus could help facilitate introduced fouling tunicates in the region, particularly in locations where additional solid substrates have created novel habitats.     

Spectrin interactions with globin chains in the presence of phosphate metabolites and hydrogen peroxide: implications for thalassaemia

We have shown the differential interactions of the erythroid skeletal protein spectrin with the globin subunits of adult haemoglobin (HbA); these indicate a preference for α-globin over that for β-globin and intact HbA in an adenosine 5′-triphosphate (ATP)-dependent manner. The presence of Mg/ATP led to an appreciable decrease in the binding affinity of the α-globin chain to spectrin and the overall yield of globin-spectrin cross-linked complexes formed in the presence of hydrogen peroxide. Similar effects were also seen in the presence of 2-,3-diphosphoglycerate (2,3 DPG), the other important phosphate metabolite of erythrocytes. The binding affinity and yield of cross-linked high molecular weight complexes (HMWCs) formed under oxidative conditions were significantly higher in α-globin compared with intact haemoglobin, HbA and the β-globin chain. The results of this study indicate a possible correlation of the preferential spectrin binding of the α-globin chain over that of the β-globin in the haemoglobin disorder β-thalassaemia.     

Grapevine leaf tissue colonization by the fungal entomopathogen <Emphasis Type="Italic">Beauveria bassiana</Emphasis><Emphasis Type="Italic">s.l.</Emphasis> and its effect against downy mildew

A study was conducted to examine whether Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) can colonize grapevine leaf tissues and subsequently confer protection against downy mildew caused by Plasmopara viticola (Berk. and Curt.) Berl. and de Toni. Following the foliar inoculation of plants with conidial suspensions of selected B. bassiana strains, colonization of leaves by the fungus was determined using culture-based and PCR techniques at different time intervals. Seven days following B. bassiana inoculation, grapevine plants were challenged with P. viticola and symptoms were assessed by calculating the disease incidence and severity. Although all tested strains were able to colonize grapevine plants, percent colonization differed significantly among strains. Disease incidence and severity were, on the other hand, significantly reduced in B. bassiana-inoculated plants compared to control plants irrespective of strain. This study is one of very few studies investigating the promising role B. bassiana could play as a plant disease antagonist.     

New furano- and pyrrolo[2,3-<Emphasis Type="Italic">d</Emphasis>]pyrimidine nucleosides and their 5′-<Emphasis Type="Italic">O</Emphasis>-triphosphates: Synthesis and biological properties

Bicyclic furano[2,3-d]pyrimidine ribonucleosides were synthesized by Pd(0)-and CuI-catalyzed coupling of 5-iodouridine with terminal alkynes. The treatment of the resulting nucleosides with ammonia or methylamine solution in aqueous alcohol resulted in pyrrolo-and N ⁷-methylpyrrolo[2,3-d]pyrimidine nucleosides. 5′-O-Triphosphates of bicyclic nucleosides were obtained by the treatment of the nucleosides with POCl₃ in the presence of a “proton sponge.” The 5′-O-triphosphates are not substrates for HCV RNA-dependent RNA polymerase, but are effective substrates for HCV RNA helicase/NTPase and did not inhibit ATP hydrolysis. Only 3-(β-D-ribofuranosyl)-6-decyl-2,3-dihydrofuro-[2,3-d]pyrimidin-2-one showed a moderate anti-HCV activity in the HCV replicon system and efficiently inhibited replication of bovine viral diarrhea virus (BVDV) in KCT-cells, other compounds being inactive. None of the compounds were cytotoxic within the tested range of concentrations.     

Pollen analysis of spider web samples from Korba District,Chhattisgarh (central India): an aerobiological aspect

Pollen analyses of spider web samples, collected from the recently planted Tectona grandis (T. grandis, teak)-dominated tropical deciduous forest of Bhulsidih Village, Korba District (Chhattisgarh, central India), shed light on the relationship between the extant vegetation and pollen rain. The study revealed the dominance of pollen of herbs and trees, whereas shrubs, fern spores and algal remains are meagre. Among the tree taxa, Sapotaceae, Syzygium, Holoptelea, Lannea coromandelica, Shorea robusta and Grewia are dominating with moderate to low and intermittent presence of Madhuca indica, Terminalia, Mitragyna, Schleichera, Anacardiaceae, Diospyros, Emblica officinalis and Flacourtia. However, the rest of the forest constituents are either not represented at all despite their presence in the floristics, which could be attributed to their low pollen productivity owing to entomophily as well as their poor pollen preservation pattern. On the other hand, the ground vegetation is represented by the very high frequency of grasses (Poaceae) along with Tubuliflorae, Chenopodiaceae/Amaranthaceae and Cerealia, however, Artemisia, Xanthium, Malvaceae, Caryophyllaceae and Justicia in moderate to lower values. Ferns, which occur abundantly along the adjoining stream banks, are marked by the sporadic retrieval of trilete spores that could be ascribed to the prevailing damp condition around the sampling provenance. The study, in addition to understanding the pollen–vegetation relationship, could also be helpful in aerobiological study, especially in assessing the allergenicity of various pollen grains/spores in the area of investigation, causing bronchial asthma, hay fever (allergic rhinitis/pollinosis), naso-bronchial allergy and other respiratory disorders along with conjunctivitis, contact dermatitis, eczema, food allergies and other health disorders.     

Comparative evaluation of <Emphasis Type="Italic">Populus</Emphasis> variants total sugar release and structural features following pretreatment and digestion by two distinct biological systems

Background

Populus natural variants have been shown to realize a broad range of sugar yields during saccharification, however, the structural features responsible for higher sugar release from natural variants are not clear. In addition, the sugar release patterns resulting from digestion with two distinct biological systems, fungal enzymes and Clostridium thermocellum, have yet to be evaluated and compared. This study evaluates the effect of structural features of three natural variant Populus lines, which includes the line BESC standard, with respect to the overall process of sugar release for two different biological systems.

Results

Populus natural variants, SKWE 24-2 and BESC 876, showed higher sugar release from hydrothermal pretreatment combined with either enzymatic hydrolysis or Clostridium thermocellum fermentation compared to the Populus natural variant, BESC standard. However, C. thermocellum outperformed the fungal cellulases yielding 96.0, 95.5, and 85.9% glucan plus xylan release from SKWE 24-2, BESC 876, and BESC standard, respectively. Among the feedstock properties evaluated, cellulose accessibility and glycome profiling provided insights into factors that govern differences in sugar release between the low recalcitrant lines and the BESC standard line. However, because this distinction was more apparent in the solids after pretreatment than in the untreated biomass, pretreatment was necessary to differentiate recalcitrance among Populus lines. Glycome profiling analysis showed that SKWE 24-2 contained the most loosely bound cell wall glycans, followed by BESC 876, and BESC standard. Additionally, lower molecular weight lignin may be favorable for effective hydrolysis, since C. thermocellum reduced lignin molecular weight more than fungal enzymes across all Populus lines.

Conclusions

Low recalcitrant Populus natural variants, SKWE 24-2 and BESC 876, showed higher sugar yields than BESC standard when hydrothermal pretreatment was combined with biological digestion. However, C. thermocellum was determined to be a more robust and effective biological catalyst than a commercial fungal cellulase cocktail. As anticipated, recalcitrance was not readily predicted through analytical methods that determined structural properties alone. However, combining structural analysis with pretreatment enabled the identification of attributes that govern recalcitrance, namely cellulose accessibility, xylan content in the pretreated solids, and non-cellulosic glycan extractability.

     

The reaction distance of a planktivorous fish (<Emphasis Type="Italic">Scardinius erythrophthalmus</Emphasis>) and the evasiveness of its prey (<Emphasis Type="Italic">Daphnia pulex </Emphasis>×<Emphasis Type="Italic"> pulicaria</Emphasis>) under different artificial light spectra

Artificial light at night may affect mortality risk in prey from visually oriented predators because the effect of the artificial light spectrum may differ for a predator’s visual prey detection and for prey evasiveness. To test this, we conducted two types of experiment. First, we assessed the reaction distance and swimming speed of juvenile rudd (Scardinius erythrophthalmus) allowed to forage on juvenile Daphnia pulex?×?pulicaria under three artificial light sources: halogen, high pressure sodium (HPS), and metal halide bulbs, at the same light intensity. Second, we assessed the evasiveness of D. pulex?×?pulicaria under the same artificial light sources and in darkness (as a control), in the presence and absence of chemical information on predation risk (kairomones) of juvenile rudd. We found that while both reaction distance and swimming speed of fish was greater under halogen compared to HPS, and similar under metal halide light compared to halogen and HPS, the evasiveness of Daphnia was greater under halogen and HPS-generated light than under metal halide light. The results suggest a possible mismatch of Daphnia’s behavioural response under metal halide light to predicted predation risk, and thus a possible threat to predator–prey balance in a lake ecosystem.