Molecular phylogeny,pigment composition,toxicology and life history of Pseudochattonella cf. verruculosa (Class Dictyochophyceae) from Wellington Harbour,New Zealand

Pseudochattonella verruculosa is a heterokont flagellate and has frequently been found associated with multi-species harmful algal blooms in Wellington Harbour. In this study the partial sequences of the nuclear encoded LSU rDNA and the large subunit of ribulose bisphosphate carboxylase (rbcL) of Pseudochattonella isolated from Wellington Harbour indicate that it is similar to P. verruculosa, while sequences of mitochondrial encoded COI, are similar to those of Pseudochattonella farcimen. As with P. farcimen, the Wellington Pseudochattonella lacked violaxanthin, lutein and anteroxanthin, three pigments detected only in P. verruculosa. The Wellington isolate also contains zeaxanthin which is absent in P. farcimen. Among all Pseudochattonella, cells of the Wellington isolate are the most variable in terms of both size and shape. Mucocysts of the Wellington Pseudochattonella also have the greatest degree of variation – from small, ‘bullet’-shape to large oval, oblong or ‘sausage’-like. In the sexual reproduction phase two gametes of the Wellington isolate fuse to form a zygote which gives rise to a large multi-nucleate cell. At times two or more of these large multi-nucleate cells fuse further to form a ‘massive’, plasmodium-like aggregate (up to 200 μm long). Positive feeding and toxicity tests on rotifers confirmed that the Wellington Pseudochattonella is cytotoxic and probably also contributed to the May 2010 fish kills. As molecular phylogenies do not conclusively support the separation of the Wellington Harbour Pseudochattonella from P. verruculosa or P. farcimen, it is tentatively named as Pseudochattonella cf. verruculosa.     

The effect of cytochalasin D on protein synthesis in Xenopus laevis oocytes

Defolliculated fully grown oocytes of Xenopus laevis were treated with cytochalasin D (10 micrograms/ml) and their protein synthesis was studied by labelling with S-35 methionine. This treatment brought about an alteration in pigment pattern as well as a reduction in amino acid uptake by the oocytes. However, the radioactive amino acid taken by cytochalasin-treated oocytes was incorporated into protein in the same proportion as in untreated oocytes. These results suggested that subcortical pigment distribution and amino acid uptake in fully grown oocytes were microfilament-dependent processes, whereas protein synthesis in the oocyte was not.     

Changes in the ultrastructure of Nematospiroides dubius (Nematoda) intestinal cells during development from fourth stage to adult

Summary The general fine structure of intestinal cells and changes which occur in ultrastructure during development from fourth-stage to adult N. dubius are reported. In fourth-stage worms pigment granules are prominent in intestinal cells. In adults the number of pigment granules appears to be reduced and phagolysosomes containing membranous profiles and pigment material increase in number. Another reorganization of cell structure involves mitochondria which are randomly distributed in the cytoplasm of cells in fourth-stage worms, concentrated in the apical cytoplasm in worms in the molting process, and confined to the base of cells in adult worms. Other changes involved structure of the nucleus, rough endoplasmic reticulum and glycogen content of cells.This investigation was supported, in part, by NIH Fellowships I-FI-GM-32750 and 5-F02-AI-32750.     

Scanning electron microscopy of the bullfrog's retina and pigment epithelium

Summary The retina and pigment epithelium of the bullfrog (Rana catesbiana) were studied with the scanning electron microscope. Fixed-dehydrated tissues were critical point dried with CO₂, then cracked in the plane of the long axis of the photoreceptors. The cellular layers of the retina and the lateral surfaces of pigment epithelial cells were visualized. The four major types of frog photoreceptor were identified: red rod, green rod, single cone, and double cone. Cone myoids were observed to be contracted in light-adapted retinas and elongated in more dark adapted retinas.This work was supported by a career development award EY-18,083 to the author and research grant EY 00468 to Dr. Kenneth T. Brown.The author gratefully acknowledges the skillful technical assistance of Ms. Maria T. Maglio.     

The fine structure and photosynthetic cost of structural leaf variegation

The leaves of some plants display an optical patchiness on their upper side, displaying light- and dark-green areas with high and low reflectance, respectively. In this investigation, we studied the fine structure of the corresponding sectors and we asked whether the lost reflected light entails a photosynthetic cost to these leaves. Four species, i.e. Arum italicum, Ranunculus ficaria, Cyclamen hederifolium and Cyclamen persicum were investigated. Scanning electron microscope examination revealed that epidermal cells of light-green sectors of all species are more bulgy than corresponding cells of neighboring dark-green leaf sectors. The comparative anatomical study revealed that (i) epidermis thickness of the light-green areas and the number of mesophyll cell layers does not differ from those of the adjacent dark-green leaf sectors and (ii) palisade cells of light-green sectors are slightly larger and more loosely arranged, allowing a much higher percentage of intercellular air spaces. The latter histological feature seems to provide the structural basis for the different optical properties between the two leaf sectors. Contrary to expectations, net photosynthetic rates (expressed on a leaf area basis) were similar in the light-green and the dark-green areas of the two cyclamen species. Yet, in C. persicum net photosynthesis was higher in the light-green areas, if expressed on a dry mass basis. The small size of the light-green spots in the rest of the test plants precluded CO₂ assimilation measurements, yet maximum linear photosynthetic electron transport rates displayed no differences between the two sectors in all plants. Thus, the assumption of a photosynthetic cost in the light-green areas was not confirmed. On the contrary, a higher construction cost was evident in the dark-green areas of three species, displaying a significantly higher specific leaf mass, without any photosynthetic benefit. The results on net photosynthesis were compatible with leaf optical properties and pigment levels. Thus, in spite of the considerably higher reflectance of the light-green areas and their lower (yet normal for a green leaf) chlorophyll levels, corresponding differences in absorptance were slight. In addition, dry mass-based pigment contents in dark-green areas were higher, while chlorophyll a/b (in two species) and carotenoid/chlorophyll ratios (in three species) were lower, pointing to a shade adaptation in these sectors. We conclude that in variegated leaves of this kind, dark-green areas are more costly to build and probably less photosynthetically active. We argue that the high pigment contents of dark-green areas establish steep light gradients in the corresponding mesophyll, rendering deeper chloroplast layers more shade adapted.     

花色豚鼠与白化豚鼠静脉血电解质、酸碱平衡及血气的比较学研究

目的比较研究花色豚鼠与白化豚鼠静脉血电解质、酸碱平衡及血气情况。方法分别取健康成年花色豚鼠和白化豚鼠,利用便携式多功能麻醉机经异氟烷吸入麻醉后,腹主静脉取血,经NOVA血气．电解质分析仪全自动分析测定电解质、酸碱平衡及血气等指标。结果花色豚鼠的Cl-、pH、SBC、PO2、SV O2、O2ct均显著低于白化豚鼠（P〈0．05,P〈0．01）,而PCO2显著高于白化豚鼠（P〈O．05）。结论花色豚鼠的携氧能力明显低于白化豚鼠,可以作为血瘀证和亚健康模型研究较好的实验动物。     

Differential microbial communities in hot spring mats from Western Thailand

The microbial communities of freshwater hot spring mats from Boekleung (Western Thailand) were studied. Temperatures ranged from over 50 up to 57°C. Green-, red-, and yellow colored mat layers were analyzed. In order to detect the major components of the microbial communities constituting the mat as well as the microorganisms showing significant metabolic activity, samples were analyzed using DNA- and RNA-based molecular techniques, respectively. Microbial community fingerprints, performed by denaturing gradient gel electrophoresis (DGGE), revealed clear differences among mat layers. Thermophilic phototrophic microorganisms, Cyanobacteria and Chloroflexi, constituted the major groups in these communities (on average 65 and 51% from DNA and RNA analyses, respectively). Other bacteria detected in the mat were Bacteroidetes, members of the Candidate Division OP10, Actinobacteria, and Planctomycetes. Differently colored mat layers showed characteristic bacterial communities and the major components of the metabolically active fraction of these communities have been identified.     

Pigment organization in the photosynthetic apparatus of Roseiflexus castenholzii

The light-harvesting-reaction center (LHRC) complex from the chlorosome-lacking filamentous anoxygenic phototroph (FAP), Roseiflexus castenholzii (R. castenholzii) was purified and characterized for overall pigment organization. The LHRC is a single complex that is comprised of light harvesting (LH) and reaction center (RC) polypeptides as well as an attached c-type cytochrome. The dominant carotenoid found in the LHRC is keto-γ-carotene, which transfers excitation to the long wavelength antenna band with 35% efficiency. Linear dichroism and fluorescence polarization measurements indicate that the long wavelength antenna pigments absorbing around 880 nm are perpendicular to the membrane plane, with the corresponding Q_y transition dipoles in the plane of the membrane. The antenna pigments absorbing around 800 nm, as well as the bound carotenoid, are oriented at a large angle with respect to the membrane. The antenna pigments spectroscopically resemble the well-studied LH2 complex from purple bacteria, however the close association with the RC makes the light harvesting component of this complex functionally more like LH1.     

CFH, VEGF, and PEDF genotypes and the response to intravitreous injection of bevacizumab for the treatment of age-related macular degeneration

We determined whether there is an association between complement factor H (CFH), high-temperature requirement A-1 (HTRA1), vascular endothelial growth factor (VEGF), and pigment epithelium-derived factor (PEDF) genotypes and the response to treatment with a single intravitreous injection of bevacizumab for age-related macular degeneration (AMD). Eighty-three patients with exudative AMD treated by bevacizumab injection were genotyped for three single nucleotide polymorphisms (SNPs; rs800292, rs1061170, rs1410996) in the CFH gene, a rs11200638-SNP in the HTRA1 gene, three SNPs (rs699947, rs1570360, rs2010963) in the VEGF gene, and four SNPs (rs12150053, rs12948385, rs9913583, rs1136287) in the PEDF gene using a TaqMan assay. The CT genotype (heterozygous) of CFH-rs1061170 was more frequently represented in nonresponders in vision than TT genotypes (nonrisk allele homozygous) at the time points of 1 and 3 months, while there was no CC genotype (risk allele homozygous) in our study cohort (p = 7.66 × 10⁻³, 7.83 × 10⁻³, respectively). VEGF-rs699947 was also associated with vision changes at 1 month and PEDF-rs1136287 at 3 months (p = 5.11 × 10⁻³, 2.05 × 10⁻², respectively). These variants may be utilized for genetic biomarkers to estimate visual outcomes in the response to intravitreal bevacizumab treatment for AMD.