Side reactions in the synthesis of phosphotyrosine-containing peptides

Summary A series of phosphopeptides Tyr(PO₃H₂)-Val-Pro-Xxx-Leu (Xxx=Met, Met(O), Nle, Dab or Cys), derived from the native platelet-derived growth factor- receptor (PDGF-) sequence, has been prepared to study their interaction with the src-homology 2 (SH2) domains of the p85 subunit of PI3 kinase. The phosphopeptides were synthesized using Fmoc methodology incorporating N-Boc dibenzyl-protected phosphotyrosine (Boc-Tyr[PO₃(Bzl)₂]) as the N-terminal amino acid, since the benzyl groups can be removed during resin cleavage with TFA. Only peptides containing methionine were found to exist partially as S-benzyl sulfonium salts after TFA cleavage from the resin. The desired peptide could be obtained from the S-benzyl sulfonium salt by hydrogenolysis.     

Effect of microspore stage and media on anther culture of peanut (Arachis hypogaea L.)

This study was designed to study the effects of stage of microspore development and culture medium on androgenic response in peanut (Arachis hypogaea L.). Anthers of various developmental stages were cultured for 7 days, then fixed and observed cytologically. Three sets of media, involving different basal media, growth regulators, sucrose levels and glutamine concentrations, were tested. In all experiments, the stage of development of the microspores at the time of culture was highly significant. The early uninucleate microspores stage was identified as producing the highest anther response rating. The effect of media was nonsignificant in all experiments. However, the stepwise modification of the media through the course of the study resulted in an almost 8 x increase in anther response rating. Numerically, the best media tested was N₆ basal medium with 1 mg 1^-1 NAA, 0.1 mg 1^-1 BA, 5.5% sucrose, and 3.5 g 1^-1 glutamine. While no haploids were obtained, four-nucleate cells were observed, indicating the potential in peanuts for an androgenic reponse.     

Social Dependence of Infant Slow Lorises to Learn Diet

Nocturnal lorises and pottos (Lorisinae) are among the least gregarious of primates. Mothers start to leave their infants alone during the night as early as the day of birth. However, captive studies also indicate that weaning young lorisines closely follow their mothers nearly all the time and obtain their first solid food via scrounging. Accordingly, it has been suggested that young lorisines depend on their mothers to obtain dietary information and to achieve dietary independence by watching their mothers feeding or interacting directly with their mothers over food. We tested for a social dependence on dietary learning by infants in a social network of wild slow lorises (Nycticebus coucang). The social network included one male infant, his mother, and two subadult females. The infant only took to mouth food items that were also part of the females' diet and showed concordance in the frequency of use of food patches with the females. These results contradict dietary learning by trial and error. They indicate that dietary learning by infants depends on information obtained from older conspecifics. However, the infant was never involved in direct interactions with conspecifics over food and fed mostly alone. He was not within a distance where he could see the females feeding more often than expected from the configuration and utilization of home ranges. The infant never looked at conspecifics feeding in his vicinity, which suggests that visual observation or direct interaction over food may not be the mechanisms by which information about food resources is passed from older individuals to young, but that other ways of obtaining such information are used.     

Trafficking to uncharted territory of the nuclear envelope

The nuclear envelope (NE) in eukaryotic cells serves as the physical barrier between the nucleus and cytoplasm. Until recently, mechanisms for establishing the composition of the inner nuclear membrane (INM) remained uncharted. Current findings uncover multiple pathways for trafficking of integral and peripheral INM proteins. A major route for INM protein transport occurs through the nuclear pore complexes (NPCs) with additional requirements for nuclear localization sequences, transport receptors, and Ran-GTP. Studies also reveal a putative NPC-independent vesicular pathway for NE trafficking. INM perturbations lead to changes in nuclear physiology highlighting the potential human disease impacts of continued NE discoveries.     

Separation methods applicable to the evaluation of enzyme-inhibitor and enzyme-substrate interactions

Enzymes catalyze a rich variety of metabolic transformations, and do so with very high catalytic rates under mild conditions, and with high reaction regioselectivity and stereospecificity. These characteristics make biocatalysis highly attractive from the perspectives of biotechnology, analytical chemistry, and organic synthesis. This review, containing 128 references, focuses on the use of separation techniques in the elucidation of enzyme-inhibitor and enzyme-substrate interactions. While coverage of the literature is selective, a broad perspective is maintained. Topics considered include chromatographic methods with soluble or immobilized enzymes, capillary electrophoresis, biomolecular interaction analysis tandem mass spectrometry (BIA-MS), phage and ribosomal display, and immobilized enzyme reactors (IMERs). Examples were selected to demonstrate the relevance and application of these methods for determining enzyme kinetic parameters, ranking of enzyme inhibitors, and stereoselective synthesis and separation of chiral entities.     

Effects of mitogen-activated protein kinases on nuclear protein import

ERK-2 MAP kinase activation induces inhibitory effects on nuclear protein import in vascular smooth muscle cells. The mechanism and characteristics of this effect of ERK-2 were investigated. An unusual dose-dependent effect of ERK-2 on nuclear protein import was identified. At higher concentrations (1 microg/mL) of ERK-2, nuclear protein import was stimulated, whereas lower concentrations (0.04 microg/mL) inhibited import. Intermediate concentrations exerted intermediate effects. The stimulatory and inhibitory effects at the 2 different ERK-2 concentrations were observed in both conventional, permeabilized cell assays of nuclear protein import and with in situ microinjection of smooth muscle cells. The biphasic effects of ERK-2 on import were also found for the other 2 members of the MAPK family, p38 and JNK. RanGAP was identified by structural analysis as a candidate target protein responsible for mediating the effects of ERK-2. After pretreatment with high concentrations of ERK-2, RanGAP activity was significantly increased by approximately 50%. In contrast, low concentrations of ERK-2 significantly attenuated RanGAP activity. These results demonstrate that all 3 members of the MAPK family can alter nuclear protein import in opposite directions depending upon the concentration of ERK-2 used. RanGAP represents the MAP kinase target whereby nuclear transport can be stimulated or inhibited.     

Seasonality in the occurrence of two lepidopterous olive pests in Egypt

Abstract Prays oleae Bern, (OM) and Palpita unionalis Hüb., (JM) are two of the most important pests in olive groves in Egypt. A 3‐year monitoring study using sex pheromone traps in semi‐arid and arid olive groves was performed. In the semi‐arid grove, flight pattern of the OM was the same as in other Mediterranean countries, but in dates concordating plant phenology. The moth completes three generations annually: the first flight is in March to April, the second is in May to June and the third occurs in August to October. In the arid olive grove, an interesting flight pattern was observed. First flight was always very close or overlapped with the second one with no male catches during August to October. However, eggs were present most of the season, indicating unusually high female presence and oviposition activity of the OM during the absence of males in the traps. Generally, moth densities were significantly lower in low fruiting years than in higher ones and were also lower in the arid olive grove than in the semi‐arid one. In contrast, JM males were present all season, exhibiting six to seven and three to four overlapping flight peaks in arid and semi‐arid olive groves, respectively. Moth densities were significantly higher in the arid olive grove than those in the semi‐arid one. This study shows that trapping location and fruit bearing year are characteristics that strongly affect the grove‐specific information needed to estimate correctly adult emergence and thus the timing of control measures.     

Comparison of the Bacterial Composition and Structure in Symptomatic and Asymptomatic Endodontic Infections Associated with Root-Filled Teeth Using Pyrosequencing

Residual microorganisms and/or re-infections are a major cause for root canal therapy failure. Understanding of the bacterial content could improve treatment protocols. Fifty samples from 25 symptomatic and 25 asymptomatic previously root-filled teeth were collected from Sudanese patients with periradicular lesions. Amplified 16S rRNA gene (V1-V2) variable regions were subjected to pyrosequencing (FLX 454) to determine the bacterial profile. Obtained quality-controlled sequences from forty samples were classified into 741 operational taxonomic units (OTUs) at 3% dissimilarity, 525 at 5% dissimilarity and 297 at 10% dissimilarity, approximately corresponding to species-, genus- and class levels. The most abundant phyla were: Firmicutes (29.9%), Proteobacteria (26.1%), Actinobacteria (22.72%), Bacteroidetes (13.31%) and Fusobacteria (4.55%). Symptomatic patients had more Firmicutes and Fusobacteria than asymptomatic patients, while asymptomatic patients showed more Proteobacteria and Actinobacteria. Interaction of disease status and age was observed by two-way ANOSIM. Canonical correspondence analysis for age, tooth restoration and disease status showed a correlation of disease status with the composition and prevalence of different members of the microbial community. The pyrosequencing analysis revealed a distinctly higher diversity of the microbiota compared to earlier reports. The comparison of symptomatic and asymptomatic patients showed a clear association of the composition of the bacterial community with the presence and absence of symptoms in conjunction with the patients’ age.