Selective reduction of post-selection CD8 thymocyte proliferation in IL-15Rα deficient mice

Peripheral CD8(+) T cells are defective in both IL-15 and IL-15Rα knock-out (KO) mice; however, whether IL-15/IL-15Rα deficiency has a similar effect on CD8 single-positive (SP) thymocytes remains unclear. In this study, we investigated whether the absence of IL-15 transpresentation in IL-15Rα KO mice results in a defect in thymic CD8 single positive (SP) TCR(hi) thymocytes. Comparison of CD8SP TCR(hi) thymocytes from IL-15Rα KO mice with their wild type (WT) counterparts by flow cytometry showed a significant reduction in the percentage of CD69(-) CD8SP TCR(hi) thymocytes, which represent thymic premigrants. In addition, analysis of in vivo 5-bromo-2-deoxyuridine (BrdU) incorporation demonstrated that premigrant expansion of CD8SP TCR(hi) thymocytes was reduced in IL-15Rα KO mice. The presence of IL-15 transpresentation-dependent expansion in CD8SP TCR(hi) thymocytes was assessed by culturing total thymocytes in IL-15Rα-Fc fusion protein-pre-bound plates that were pre-incubated with IL-15 to mimic IL-15 transpresentation in vitro. The results demonstrated that CD8SP thymocytes selectively outgrew other thymic subsets. The contribution of the newly divided CD8SP thymocytes to the peripheral CD8(+) T cell pool was examined using double labeling with intrathymically injected FITC and intravenously injected BrdU. A marked decrease in FITC(+) BrdU(+) CD8(+) T cells was observed in the IL-15Rα KO lymph nodes. Through these experiments, we identified an IL-15 transpresentation-dependent proliferation process selective for the mature CD8SP premigrant subpopulation. Importantly, this process may contribute to the maintenance of the normal peripheral CD8(+) T cell pool.     

Immunolocalization of 8-5′ and 8-8′ linked structures of lignin in cell walls of Chamaecyparis obtusa using monoclonal antibodies

Mouse monoclonal antibodies were generated against dehydrodiconiferyl alcohol- or pinoresinol-p-aminohippuric acid (pAHA)-bovine serum albumin (BSA) conjugate as probes that specifically react with 8-5′ or 8-8′ linked structure of lignin in plant cell walls. Hybridoma clones were selected that produced antibodies that positively reacted with dehydrodiconiferyl alcohol- or pinoresinol-pAHA–BSA and negatively reacted with pAHA–BSA and guaiacylglycerol-beta-guaiacyl ether-pAHA–BSA conjugates containing 8-O-4′ linkage. Eight clones were established for each antigen and one of each clone that positively reacted with wood sections was selected. The specificity of these antibodies was examined by competitive ELISA tests using various lignin dimers with different linkages. The anti-dehydrodiconiferyl alcohol antibody reacted specifically with dehydrodiconiferyl alcohol and did not react with other model compounds containing 8-O-4′, 8-8′, or 5-5′ linkages. The anti-pinoresinol antibody reacted specifically with pinoresinol and syringaresinol and did not react with the other model compounds containing 8-O-4′, 8-5′, or 5-5′ linkages. The antibodies also did not react with dehydrodiconiferyl alcohol acetate or pinoresinol acetate, indicating that the presence of free phenolic or aliphatic hydroxyl group was an important factor in their reactivity. In sections of Japanese cypress (Chamaecyparis obtusa), labeling by the anti-dehydrodiconiferyl alcohol antibody was found in the secondary walls of phloem fibers and in the compound middle lamellae, and secondary walls of tracheids. Weak labeling by the anti-pinoresinol antibody was found in secondary walls of phloem fibers and secondary walls and compound middle lamellae of developed tracheids. These labelings show the localization of 8-5′ and 8-8′ linked structure of lignin in the cell walls.     

Population density of the Réunion Grey White-eye Zosterops borbonicus within the summit ecosystems of Réunion,Mascarene Islands

Assessing population density is crucial for studying the ecology and evolutionary biology of species as well as for conservation purposes. Here we used point count methods to infer population density in a single-island endemic passerine bird, the Réunion Grey White-eye Zosterops borbonicus, that displays striking evidence of differentiation at a small spatial scale. Population density was estimated at 5.17 birds ha^?1 (CL 4.85–5.50), a value somewhat higher than previously believed. This estimation provides the first detailed estimation of bird population density in the vulnerable summit ecosystems of Réunion and will possibly allow a better understanding of the evolutionary causes of this plumage colour variation.     

Absolute configuration of 2′(R)-acetylmontrifoline and 2′(R)-montrifoline,furoquinolines from the fruits of Teclea nobilis

A phytochemical study on the alkaloids from Teclea nobilis yielded five furoquinolines, including the new acetylmontrifoline. The structures were elucidated from extensive 2D-NMR and MS experiments. The absolute configuration of the chiral center of montrifoline and of acetylmontrifoline was established as R by analysing the Mosher's derivatives and chemical modifications, respectively. All the five furoquinolines showed weak in vitro antiplasmodial activity against Plasmodium falciparum, montrifoline being moderately active (56 μg/ml) and not cytotoxic.     

Genomic organization of human complement protein C8α and further examination of its linkage to C8β

Human C8 is one of five complement components (C5b, C6, C7, C8, C9) that interact to form the cytolytic C5b-9 complex on target membranes. It is composed of three nonidentical subunits (C8, C8, C8) encoded by separate genes. C8 and C8 are linked on chromosome 1p32, whereas C8 is located on 9q22.3-q32. In this study, overlapping genomic clones were isolated and used to decipher the organization of the human C8 gene. The gene contains at least 11 exons spanning 70kb of DNA. When compared to C6, C8 and C9, there is a remarkable similarity in genomic organization, consistent with amino acid sequence comparisons that suggest these proteins are ancestrally related. Regions of each protein that are structurally similar are encoded in exons of correspondingly similar lengths with highly conserved boundaries and phases. Availability of genomic sequence also facilitated a more detailed analysis of C8 and C8 linkage. Based on analysis of genomic digests with cDNA probes, the loci were previously reported to be physically linked (< 2.5kb) and in a 5- 3 orientation. In the present study, results obtained using exon-specific probes indicate the loci are not as closely linked as initially believed. Furthermore, they suggest that cDNA probes used earlier yielded misleading information because they encode exons that are distributed across large segments of genomic DNA.     

Direct connections between the R7/8 and R1–6 photoreceptor subsystems in the dipteran visual system

Summary Musca and related flies have three main photoreceptor subsystems. The R1–6 group has short axons that terminate in the cartridges of the first optic neuropile, the lamina. The cartridges are bypassed by the longer axons of R7 and R8, which run together to terminate at different levels in the underlying medulla neuropile. The present account describes a shallow, previously unidentified zone in the lamina within which R7/8 make glancing contact with R1–6. At the distal border of the cartridge over no more than 3–4 m depth, the tangentially directed short axon of R6 squeezes between the pair from R7 and R8, forming quite large areas of mutual contact (approximately 7 m²). Less frequently, R1 is contacted. At least some of these sites contain smaller membrane specialisations indistinguishable from the more numerous gap junctions found more proximally that interconnect the terminals of R1–6. The R7/8 junctions with R6 are of comparable size (0.15 m²) and likewise possess symmetrical membrane densities. They provide proposed pathways for direct electrical interaction to account for observed electrical input from R7/8 to the R1–6 subsystem. In two cases R7/8 was possibly postsynaptic to R1–6 at a multiple-contact synapse, but even if functional, these sites were so rare that they are unlikely to have much operational significance.     

8-Aza Analogues of Deaza Purine Nucleosides. Synthesis and Biological Evaluation of 8-Aza-1-deazaadenosine and 2′-Deoxy-8-aza-1-deazaadenosine

Abstract

The syntheses of 7-amino-3-(β-D-ribofuranosyl)-3H-1,2,3-triazolo[4,5-b]pyridine (8-aza-1-deazaadenosine) (2) and 7-amino-3-(2-deoxy-β-D-erythro-pentofuranosyl)-3H-1,2,3-triazolo[4,5-b]pyridine (2′-deoxy-8-aza-1-deazaadenosine) (3) by glycosylation of the anion of 7-chloro-3H-1,2,3-triazolo[4,5-b]pyridine are described. The anomeric configuration as well as the position of glycosylation were determined by ¹H, ¹³ NMR, UV and N.O.E. difference spectroscopy. The cytotoxicity of these nucleosides against several murine and human tumor cell lines is discussed. Compounds 2 and 3 proved to be good inhibitors of adenosine deaminase.     

Pollen analysis of honeys from the central zone of the Argentine province of Entre Ríos

Based on the melissopalynology 38 honey samples collected in the central region of the Argentine province of Entre Ríos were classified by botanical and geographical origin. According to qualitative analysis, 20 honey samples were monofloral and 18 were multifloral. Dominant pollen types were Scutia buxifolia Reissek (Rhamnaceae) in six samples, Baccharis spp. (Asteraceae) in five, Lotus spp. (Fabaceae) in three, Eucalyptus spp. (Myrtaceae) and Eryngium spp. (Apiaceae) in two, Ammi visnaga (L.) Lam. (Apiaceae) and Trithrinax campestris (Burmeist.) Drude & Griseb. (Arecaceae) in one sample. One hundred and nineteen pollen types were identified belonging to 52 families; 75% of which were native species. The families best represented in number of species were Asteraceae and Fabaceae. Pollen types such as Scutia buxifolia, Trithrinax campestris, Schinus spp. (Anacardiaceae), Mimosoideae from Prosopis spp., Acacia spp., Mimosa ostenii Speg. ex Burkart, and M. strigillosa Torr. & A. Gray are considered the indicators for this geographical origin. The studied honeys were also characterized by a high frequency of Apiaceae, Brassicaceae, Astereae, Echium plantagineum L. and cultivated Papilionoideae forage species such as Melilotus albus Desr., Lotus spp. and Trifolium spp. Honeydew elements were scarce.     

Rôle of phagostimulants of cotton leaves in the feeding behaviour of Spodoptera littoralis

Petroleum ether extract of cotton leaves was the most attractive to the hatched larvae of Spodoptera littoralis, and it remained so even after removal of the chlorophyll and other pigments. Steam distillation of this filtrate gave a volatile oil which also proved highly attractive to the larvae. In this fraction, 12 components were identified by thin-layer chromatography. Of these components, α-terpineol, citronellol and α-pinene were the most attractive, whereas humulene and linalool were less attractive. The remaining non-volatile lipid fraction was also attractive to the larva. In this fraction the unsaponifiable matter contained the attractive ingredients. This fraction proved to be a hydrocarbon. The infrared spectrum and nuclear magnetic resonance showed only typical hydrocarbon bonds.     

Spring movements of Rüppell's Vulture Gyps rueppellii across the Strait of Gibraltar

Most of our understanding of territorial behaviour in birds comes from studies of temperate species. Territoriality and factors influencing territory size of tropical birds are still poorly studied. In this study, 11 territories of the Cinnamon-breasted Rock Bunting Emberiza tahapisi were measured in the Amurum Forest Reserve and adjoining farmlands. Factors influencing the habitat choice of this species were also studied. Territory sizes ranged from 0.1–0.3 ha with a mean territory size of 0.19 ± 0.1ha. Territory size decreased with increasing food abundance and increased with tree height and number of saplings. The birds did not show signs of territoriality in farmlands but flocked in small groups (mean group size 6.88 ± 0.89). Flock size also increased with increasing number of seeds. In the forested land, birds showed a preference for rocky outcrops, whereas in farmlands, they appeared to prefer open and newly cultivated farmlands with little or no ground cover. Understanding the influence of habitat variables on territory size and habitat selection in a locally abundant and widely distributed tropical species such as the Cinnamon-breasted Rock Bunting thus offers unique opportunities for understanding how changes in landscape caused by land-use regimes may affect populations.     

Degradation of Cell Wall of Mucor mucedo by Chitosanase from Bacillus R–4

To broaden our understanding of extracellular proteins of Aspergillus oryzae at the conidial germination stage, analyses of the secreted proteins during germination were carried out. Taka-amylase A (TAA), glucoamylase (GLAA), and aspergillopepsin A (PEPA) were identified as the main products by peptide mass fingerprinting. TAA and PEPA were detected simultaneously with the formation of germ tubes. With the development of germination, the pH of the medium fell from 5.5 to 3.5. The secreted PEPA had a pro-sequence and likely shifted from 42 kDa to 41 kDa below pH 4.6, indicating that the precursor of PEPA was secreted and underwent pH-dependent processing. Furthermore, the 41 kDa protein was trapped by the addition of pepstatin A, the specific inhibitor of PEPA, suggesting that the maturation of pro-PEPA was a stepwise autoprocessing upon acidification of the medium and itself was an intermediate of the processing. It was implied that PEPA plays an important role at the early germination stage.     

Synthesis and neuropharmacological evaluation of esters of R(−)-N-alkyl-11-hydroxy-2-methoxynoraporphines

We synthesized several esters of R(?)-N-alkyl-11-hydroxy-2-methoxynoraporphines, assessed their affinities at dopamine D₁ and D₂ receptors in rat forebrain tissue and quantified their effects on motor activity in normal adult male rats. Tested compounds displayed moderate to high affinities to D₂ receptors but low affinities to D₁ receptors. The most D₂-potent (K_i = 18.9 nM) and selective novel agent (>529-fold vs D₁ sites) was R(?)-2-methoxy-11-acetyloxy-N-n-propylnoraporphine (compound 4b). At moderate doses, the compound proved to have prolonged behavioral locomotor activity.     

扩展青霉脂肪酶ep8与R182K叠加突变体的构建

利用重叠延伸PCR法对扩展青霉碱性脂肪酶(PEL)基因进行体外定点突变,并将含突变基因的重组质粒pAO815-ep8-R182K在毕赤酵母(Pichiapastoris)GS115中进行表达。叠加突变体PEL-ep8-R182K表达产物与野生型PEL、PEL-ep8比较实验表明:叠加突变体表达蛋白PEL-ep8-R182K最适反应温度与野生型PEL、PEL-ep8一致,均为40℃;热稳定性与野生型相似,比PEL-ep8降低2.25℃。但是,在比活上,PEL-ep8-R182K与PEL-ep8、野生型PEL相比,其比酶活分别提高了14.03%和3.86%。