Micromorphology of Scirpus (Cyperaceae) and related genera in south-west Europe

Glume surface, fruit surface, perianth bristles and pollen morphology in Scirpus L. and related genera from south-west Europe have been investigated by light and scanning electron microscopy. The results of this analysis confirm the heterogeneity of the group and provide further support to the current recognition of the genera Scirpus L., Bolboschoenus (Rchb.) Palla , Scirpoides Ség., Schoenoplectus (Rchb.) Palla, Isolepis R.Br., Trichophorum Pers. and Blysmus Panz. as well as to a re-appraisal of subg. Actaeogeton (Rchb.) Oteng-Yeb. of Schoenoplectus .  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 145 , 45–58.     

Effects of light (photoperiod,spectral composition) on the population dynamics of Tisbe holothuriae Humes (Copepods,Harpacticoida)

The harpacticoid copepod Tisbe holothuriae Humes was reared for several generations under different photoperiods (white light) and spectral quality (continuous illumination). The main life cycle parameters were measured and demographic variables were determined. Development time was retarded under green wavelengths and at photoperiods different from LD 12:12, especially under continuous white light. Total body length of females and males was shortest under LD 12:12 and longest under constant dark. Photoperiods different from LD 12:12 prolonged the interval between broods of egg sacs. Short photoperiods caused higher abortion rate, while long photoperiods retarded the maturation time of the egg sacs. Red wavelengths stimulated the abortion rate. Photoperiod LD 12:12 and blue wavelengths increased the production of offspring and the life span of females. Highest intrinsic rates of natural increase were estimated under LD 12:12 (rm =0.304) and blue wavelengths (rm = 0.254). These light conditions proved the most favourable for rearing Tisbe holothuriae.     

The chalcone synthase multigene family of Petunia hybrida (V30): differential,light-regulated expression during flower development and UV light induction

We have analysed the expression of the 8–10 members of the gene family encoding the flavonoid biosynthetic enzyme chalcone synthase (CHS) from Petunia hybrida. During normal plant development only two members of the gene family (CHS-A and CHS-J) are expressed. Their expression is restricted to floral tissues mainly. About 90% of the total CHS mRNA pool is transcribed from CHS-A, wheares CHS-J delivers about 10% in flower corolla, tube and anthers. Expression of CHS-A and CHS-J during flower development is coordinated and (red) light-dependent. In young seedlings and cell suspension cultures expression of CHS-A and CHS-J can be induced with UV light. In addition to CHS-A and CHS-J, expression of another two CHS genes (CHS-B and CHS-G) is induced in young seedlings by UV light, albeit at a low level. In contrast to CHS genes from Leguminoseae, Petunia CHS genes are not inducible by phytopathogen-derived elicitors. Expression of CHS-A and CHS-J is reduced to a similar extent in a regulatory CHS mutant, Petunia hybrida Red Star, suggesting that both genes are regulated by the same trans-acting factors. Comparison of the promoter sequences of CHS-A and CHS-J reveals some striking homologies, which might represent cis-acting regulatory sequences.     

The tricks plants use to reach appropriate light

The perception of ambient light signals that produce a relevant response to ensure exposure to appropriate levels of light energy is vital for plants. In response to this, intricate molecular mechanisms to mediate light signaling have evolved in plants. Among the responses induced by light, seedling extension is a determining event for plant survival in darkness, especially in the initial stage of plant growth. Here we review previous studies and recent progress towards an understanding of light signaling that regulates seedling elongation. We focus on the three regions of the sunlight spectrum that primarily control seedling elongation, namely red/far-red light, blue/UV-A light and UV-B light, and summarize the four signaling pathways that correspond to the three effective spectra.     

Photo-induced unfolding and inactivation of bovine carbonic anhydrase in the presence of a photoresponsive surfactant

Photoreversible changes in the conformation and enzymatic activity of bovine carbonic anhydrase have been investigated as a function of photoresponsive surfactant concentration and light conditions. The light-responsive surfactant undergoes a photoisomerization from the relatively hydrophobic trans isomer under visible light to the relatively hydrophilic cis isomer upon UV illumination, providing a means to photoreversibly control enzyme–surfactant interactions. Small-angle neutron scattering and dynamic light scattering measurements, along with fluorescence spectroscopy, indicate that carbonic anhydrase unfolds upon addition of the surfactant under visible light, while only a small degree of unfolding is observed under UV light. Therefore, the enzyme is completely inactivated in the presence of the trans surfactant, while 40% of the native activity is preserved under UV light, providing a photoreversible “on/off switch” of enzyme activity. Small-angle neutron scattering data provide details of the in vitro conformational changes of the enzyme in response to the photosurfactant and light, with the enzyme found to aggregate as a result of photosurfactant-induced unfolding. Fourier transform infrared (FT-IR) spectroscopy further provides information on the secondary structure changes of the protein in the presence of photosurfactant.     

Modulation of in vivo IgG crystallization in the secretory pathway by heavy chain isotype class switching and N-linked glycosylation

Crystalline bodies (CBs) can develop in the endoplasmic reticulum (ER) of antibody-producing cells. Although this phenotype is often reported in association with plasma cell dyscrasias and other hematological disorders, the details of CB biogenesis and CB's roles in pathophysiology remain poorly understood. Using an imaging-based screening method, we identified a secretion-competent human IgG2/λ clone that develops spindle-shaped intracellular crystals in transiently-transfected HEK293 cells upon Brefeldin A treatment. When stably overexpressed from CHO cells, the IgG2/λ clone spontaneously produced spindle-shaped CBs in the ER. Some CBs were released to the extracellular space while remaining enclosed by the membranes of secretory pathway origin. Structural modeling on the variable-region did not uncover prominent surface characteristics such as charge clusters. In contrast, alterations to the constant domain-encoded properties revealed their modulatory roles in CB-inducing propensities and CB morphology. For example, deletion of the entire Fc domain changed the morphology of CBs into thin filaments. Elimination of an N-linked glycan by a N297A mutation promoted Russell body biogenesis accompanied by marked reduction in IgG secretion. Isotype class switching from the original IgG2 to IgG1 and IgG4 changed the crystal morphology from spindle-shaped to long needle and acicular shaped, respectively. The IgG3 version, in contrast, suppressed the CB formation. Either the HC or LC alone or the Fc-domain alone did not trigger CB biogenesis. An IgG's in vivo crystal morphology and crystallization propensity can thus be modulated by the properties genetically and biochemically encoded in the HC constant region.     

Determination of propafenone hydrochloride by flow‐injection analysis coupled with resonance light scattering detection

A simple, sensitive and rapid flow injection analysis (FIA) method with resonance light scattering (RLS) was described for the determination of propafenone (PPF). The method was based on the ion‐association reaction of 12‐tungstophosphoric acid (TP) with propafenone. In pH 1.0 acidic medium, TP reacted with PPF to form an ion‐associate complex, which resulted in a significant enhancement of RLS intensity. The maximum scattering peak was located at 340 nm, the RLS intensity was proportional to the concentration of PPF in the range 0.003–9.0 µg/mL, and the detection limit (3σ) of 1.0 ng/mL was obtained at a sampling rate of 60 samples/h. The feasible reaction conditions and FIA parameters for the system were optimized. The method proposed in this paper shows satisfactory reproducibility with a relative standard deviation (RSD) of 2.1% for 10 successive determinations of 2.0 µg/mL PPF. The present method had been successfully applied to the determination of PPF in serum samples and pharmaceutical samples. The results obtained were in agreement with the method used in the Chinese Pharmacopoeia. Copyright © 2008 John Wiley & Sons, Ltd.     

Influence of quaternary cation compound on the size of the Escherichia coli small multidrug resistance protein,EmrE

EmrE is a member of the small multidrug resistance (SMR) protein family in Escherichia coli. It confers resistance to a wide variety of quaternary cation compounds (QCCs) as an efflux transporter driven by the transmembrane proton motive force. We have expressed hexahistidinyl (His₆) – myc epitope tagged EmrE, extracted it from membrane preparations using the detergent n-dodecyl-β-D-maltopyranoside (DDM), and purified it using nickel-affinity chromatography. The size of the EmrE protein, in DDM environment, was then examined in the presence and absence of a range of structurally different QCC ligands that varied in their chemical structure, charge and shape. We used dynamic light scattering and showed that the size and oligomeric state distributions are dependent on the type of QCC. We also followed changes in the Trp fluorescence and determined apparent dissociation constants (K_d). Overall, our in vitro analyses of epitope tagged EmrE demonstrated subtle but significant differences in the size distributions with different QCC ligands bound.     

DNA sequence changes in mutations induced by ultraviolet light in the gpt gene on the chromosome of Escherichia coli uvr+ and urvA cells

Summary Sequence changes in mutations induced by ultraviolet light are reported for the chromosomal Escherichia coli gpt gene in almost isogenic E. coli uvr ⁺ and excision-deficient uvrA cells. Differences between the mutagenic spectra are ascribed to preferential removal of photoproducts in the transcribed strand by excision repair in uvr ⁺ cells. This conclusion is confirmed by analysis of published results for genes in both uvr ⁺ and uvr ^– cells, showing a similar selective removal of mutagenic products from the transcribed strand of the E. coli lacI gene and of the lambda phage cl repressor gene. Comparison of these data with published results for ultraviolet mutagenesis of gpt on a chromosome in Chinese hamster ovary cells showed that a mutagenic hot spot in mammalian cells is not present in E. coli; the possibility is suggested that the hot spot might arise from localized lack of excision repair. Otherwise, mutagenesis in hamster cells appeared similar to that in E. coli uvr ⁺ cells, except there appears to be a smaller fraction of single-base additions and deletions (frameshifts) in mammalian than in bacterial cells. Phenotypes of 6-thioguanine-resistant E. coli showed there is a gene (or genes) other than gpt involved in the utilization of thioguanine by bacteria.