Orbicules in angiosperms: Morphology, function, distribution, and relation with tapetum types

Orbicules, or Ubisch bodies, are sporopollenin particles lining the inner tangential and sometimes also the radial tapetal cell walls. They occur only in species with a secretory tapetum. The surface ornamentation of orbicules and pollen of the same species is often strikingly similar. Although orbicules were discovered more than a century ago, these structures remain enigmatic since their function is still obscure. Proposed hypotheses about their possible function are discussed. We also deal here with topics such as the possible allergenicity of orbicules and their representation in the fossil record. The use of orbicule characters for systematics is reviewed. The distribution of orbicules throughout the angiosperms, based on a literature review from the first report until today, is shown in a list with 314 species from 72 families. Those species found in the literature without orbicules are presented together with their tapetum type. We plotted this information on a dahlgrenogram to visualize the distribution of orbicules. Orbicules occur in all subclasses of the angiosperms. Their occurrence is not correlated with certain modes of pollination or habitats.

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Résumé  Les orbicules, ou corps d’Ubisch, sont des particules de sporopollénine couvrant la surface intérieure tangentiale et parfois la surface radiale des cellules du tapétum. On ne les retrouve que dans les espèces possédant un tapétum sécréteur. L’ornementation superficielle des orbicules et celle du pollen d’une même espèce est souvent remarquablement similaire. Malgré le fait que les orbicules ont été découvert il y a plus d’un siècle, ces structures restent énigmatiques et leur fonction est toujours méconnue. Les hypothèses proposées concernant la fonction éventuelle des orbicules sont commentées dans cet article. Nous avons également traité des sujets tels que les éventuels effets allergènes des orbicules ainsi que leur présence dans les strates fossiles. L’utilisation de caractères orbiculaires dans la systématique est analysée. Nous présentons une liste de 314 espèces appartenant à 72 familles possédant des orbicules, sur base d’une analyse de la litérature à partir de la première observation jusqu’au présent. Pour les espèces rapportées dans la litérature qui ne possèdent pas d’orbicules, nous présentons aussi leur type de tapétum. Nous avons projeté cette information sur un Dahlgrenogramme afin de visualiser la distribution des orbicules. Nous les retrouvons dans toutes les sous-classes des angiospermes. Leur présence n’est pas correlée avec certains modes de pollinisation ou avec divers types d’habitat.

     

Synthesis of biologically active steroid derivatives by the utility of Lawesson's reagent

Steroid derivatives V, VI, VII and VIII reacted with Lawesson's reagent (LR) to produce spiro-oxazaphosphole-4',17-androstene derivative XI, diazaphospholoandrostane XIV and the thionated derivatives XVI and XVII, respectively. The structures of the new compounds were confirmed by analytical and spectroscopic evidence. A mechanism accounting for the formation of the new compounds was given. The in vitro antimicrobial activity of the new compounds were tested.     

Polyclonal antibodies against the recombinantly expressed coat protein of the Citrus psorosis virus

Psorosis is a damaging disease of citrus that is widespread in many parts of the world. Citrus psorosis virus (CPsV), the type species of the genus Ophiovirus, is the putative causal agent of psorosis. Detection of CPsV by laboratory methods, serology in particular is a primary requirement for large-scale surveys but their production has been impaired by the difficulty of obtaining sufficient clean antigen for immunization. Specific PAbs against coat protein were produced in E. coli using recombinant DNA approach. The full length CP gene fragment was amplified by RT-PCR using total RNA extracted from CPsV infected citrus leaves and CP specific primers. The obtained product (1320bp) was cloned, sequenced and sub-cloned into p^ET-30(+) expression vector. Expression was induced and screened in different bacterial clones by the presence of the expressed protein (48kDa) and optimized in one clone. Expressed CP was purified using batch chromatography under denaturing conditions. Specificity of expressed protein was demonstrated by ELISA before used as antigen for raising PAbs in mice. Specificity of the raised PAbs to CPsV was verified by ELISA and western blotting. The raised PAbs were showed highly effectiveness in screening by ELISA comparing with the commercial antibodies purchased from Agritest, Valanzano, Italy.The expression of CPsV CP gene in E. coli, production of PAbs using recombinant protein as an antigen, the suitability of these antibodies for use in immunodiagnostics against the CPsV Egyptian isolate have been accomplished in this work.     

The Differentiation of Live From Dead Sperms in Fowl Semen

A combined stain solution is made by dissolving 0.1 gm bromphenol blue and 0.2 gm nigrosin in 100 ml of a M/15 buffer solution of KH₂PO₄ and Na₂HPO₄ adjusted to pH 7.5. This staining solution was used to prepare stained fowl semen smears. Such smears give stable differentiation of live from dead sperms. The dead sperms are stained with a dark violet color while the live ones are not stained.     

Rapid cloning of any rearranged mouse immunoglobulin variable genes

Immunoglobulins (Ig) have been the focus of extensive study for several decades and have become an important research area for immunologists and molecular biologist. The use of polymerase chain reaction (PCR) technology has accelerated the cloning, sequencing, and characterization of genes of the immune system. However, cloning and sequencing the Ig variable (V) genes using the PCR technology has been a challenging task, primarily due to the very diverse nature of Ig V region genes. We have developed a simple, rapid, and reproducible PCR-based technique to clone any rearranged mouse Ig heavy or light chain genes. A close examination of all Ig heavy and light chain V gene families has resulted in the design of 5 and 3 universal primers from regions that are highly conserved across all heavy or light chain V gene families, and the joining or constant regions, respectively. We present our strategy for designing universal primers for Ig V gene families. These primers were able to rapidly amplify the rearranged Ig V genes, belonging to diverse Ig V gene families from very different cell lines, i.e., J558, MOPC-21, 36–60, and a chicken ovalbumin specific B-cell hybridoma. In addition, the present study provides the complete alignment of nucleotide sequences of all heavy and light chain variable gene families. This powerful method of cloning Ig V genes, therefore, allows rapid and precise analysis of B-cell hybridomas, B-cell repertoire, and B-cell ontogeny.The nucloetide sequence data reported in this paper have been submitted to the EMBL/GenBank nucleotide sequence database and have been assigned the accession number U32111     

Mutation of a protein kinase C phosphorylation site in the erbB protein of avian erythroblastosis virus.

Tumor promoter-stimulated phosphorylation of threonine 98 of the erbB protein of avian erythroblastosis virus (AEV) correlates with inhibition of erbB-dependent mitogenesis. To more clearly define the role of phosphorylation of this residue in regulation of the activity of the erbB protein, we have constructed erbB mutations which encode alanine (Ala-98), tyrosine (Tyr-98), or serine (Ser-98) at position 98. The biosynthesis and stability of the three mutant proteins were similar to those of the wild-type erbB protein, and all three retained the ability to transform chicken embryo fibroblasts. Treatment of transformed CEF with 12-tetradecanoylphorbol-13-acetate (TPA) stimulated incorporation of 32Pi into wild-type and mutant erbB proteins and resulted in a slight decrease in the electrophoretic mobilities of all the erbB proteins. Tryptic maps of erbB phosphopeptides showed no endogenous or TPA-stimulated phosphorylation of alanine 98 or tyrosine 98 in cells transformed by the Ala-98 and Tyr-98 mutants. Analysis of tryptic phosphopeptides by high-pressure liquid chromatography revealed that TPA treatment of cells stimulated phosphorylation of other sites of the erbB protein in addition to threonine 98. A high endogenous level of phosphorylation of serine 98 of the Ser-98 mutant protein was found, and TPA treatment of cells did not result in further phosphorylation of this residue. Cells transformed by wild-type and mutant AEV were equally sensitive to TPA-dependent inhibition of growth in soft agar and TPA-dependent inhibition of [3H]thymidine incorporation. TPA treatment inhibited tyrosine phosphorylation to a similar extent in cells transformed by wild-type or Ala-98 AEV. These data indicate that phosphorylation of threonine 98 of the erbB protein is not responsible for TPA-dependent inhibition of growth of AEV-transformed cells or TPA-induced inhibition of erbB-dependent tyrosine phosphorylation. TPA-stimulated phosphorylation of the erbB protein at other sites may mediate these effects. The data also show that subtle changes in a phosphorylation site (i.e., changing threonine to serine) can drastically alter recognition by protein kinases.     

Spectrofluorimetric determination of dopamine receptor antagonist LE 300 in mice plasma

A simple, rapid and highly sensitive spectrofluorimetric method was developed for determination of a novel type of dopamine receptor antagonist LE300 in mouse plasma. The method is based on measuring the native fluorescence of LE‐300 in methanol at 343 nm after excitation at 280 nm. The fluorescence concentration plot was rectilinear over the range of 3.5–100 ng/mL with a lower detection limit of 1.0 ng/mL and quantification limit of 3.5 ng/mL. The method was statistically validated for linearity, accuracy, precision and selectivity according to the International Conference on Harmonization guidelines. The accuracy and precision results was expressed as % recovery and relative standard deviation (RSD). The accuracy for LE‐300 was in the range 95.5–103.6% and RSD values were in the range of 0.21–1.55% of the theoretical value. The method was successfully applied to the analysis of LE‐300 in mice plasma. The results were compared statistically with those obtained by the reported method and were found to be in good agreement, which could be applied in a pharmacokinetic study. Copyright © 2015 John Wiley & Sons, Ltd.     

Impact of ozonation pre-treatment of oil sands process-affected water on the operational performance of a GAC-fluidized bed biofilm reactor

Treatment of oil sands process-affected water (OSPW) using biodegradation has the potential to be an environmentally sound approach for tailings water reclamation. This process is both economical and efficient, however, the recalcitrance of some OSPW constituents, such as naphthenic acids (NAs), require the pre-treatment of raw OSPW to improve its biodegradability. This study evaluated the treatment of OSPW using ozonation followed by fluidized bed biofilm reactor (FBBR) using granular activated carbon (GAC). Different organic and hydraulic loading rates were applied to investigate the performance of the bioreactor over 120 days. It was shown that ozonation improved the adsorption capacity of GAC for OSPW and improved biodegradation by reducing NAs cyclicity. Bioreactor treatment efficiencies were dependent on the organic loading rate (OLR), and to a lesser degree, the hydraulic loading rate (HLR). The combined ozonation, GAC adsorption, and biodegradation process removed 62 % of chemical oxygen demand (COD), 88 % of acid-extractable fraction (AEF) and 99.9 % of NAs under optimized operational conditions. Compared with a planktonic bacterial community in raw and ozonated OSPW, more diverse microbial communities were found in biofilms colonized on the surface of GAC after 120 days, with various carbon degraders found in the bioreactor including Burkholderia multivorans, Polaromonas jejuensis and Roseomonas sp.