High-frequency P element loss in Drosophila is homolog dependent

P transposable elements in Drosophila melanogaster can undergo precise loss at a rate exceeding 13% per generation. The process is similar to gene conversion in its requirement for a homolog that is wild type at the insertion site and in its reduced frequency when pairing between the homologs is inhibited. However, it differs from classical gene conversion by its high frequency, its requirement for P transposase, its unidirectionality, and its occurrence in somatic and premeiotic cells. Our results suggest a model of P element transposition in which jumps occur by a "cut-and-paste" mechanism but are followed by double-strand gap repair to restore the P element at the donor site. The results also suggest a technique for site-directed mutagenesis in Drosophila.     

入侵害虫螺旋粉虱及其在我国的适生区预测

主要介绍热带亚热带入侵害虫螺旋粉虱Aleurodicus dispersus Russell的形态特征、分布、寄主与生物学特性;利用DYMEX软件分析螺旋粉虱在我国潜在的分布区。模型分析结果表明,最适宜的潜在的适生区主要集中在中国的南部,包括海南、广东、广西以及台湾等的大部分区域;在云南只有零散的潜在适生区;而广大的西北和东北不利于该虫的生存;依据适生性风险预测结果,提出了应急处理措施。     

The role of posttranslational modification in moonlighting glyceraldehyde-3-phosphate dehydrogenase structure and function

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a moonlighting protein exhibiting distinct activities apart from its classical role in glycolysis. Regulation of its moonlighting functions and its subcellular localization may be dependent on its posttranslational modification (PTM). The latter include its phosphorylation, which is required for its role in intermembrane trafficking, synaptic transmission and cancer survival; nitrosylation, which is required for its function in apoptosis, heme metabolism and the immune response; acetylation which is necessary for its modulation of apoptotic gene regulation; and N-acetylglucosamine modification which may induce changes in GAPDH oligomeric structure. These findings suggest a structure function relationship between GAPDH posttranslational modification and its diverse moonlighting activities.

     

Phosphorylation Regulates the Ubiquitin-independent Degradation of Yeast Pah1 Phosphatidate Phosphatase by the 20S Proteasome

Saccharomyces cerevisiae Pah1 phosphatidate phosphatase, which catalyzes the conversion of phosphatidate to diacylglycerol for triacylglycerol synthesis and simultaneously controls phosphatidate levels for phospholipid synthesis, is subject to the proteasome-mediated degradation in the stationary phase of growth. In this study, we examined the mechanism for its degradation using purified Pah1 and isolated proteasomes. Pah1 expressed in S. cerevisiae or Escherichia coli was not degraded by the 26S proteasome, but by its catalytic 20S core particle, indicating that its degradation is ubiquitin-independent. The degradation of Pah1 by the 20S proteasome was dependent on time and proteasome concentration at the pH optimum of 7.0. The 20S proteasomal degradation was conserved for human lipin 1 phosphatidate phosphatase. The degradation analysis using Pah1 truncations and its fusion with GFP indicated that proteolysis initiates at the N- and C-terminal unfolded regions. The folded region of Pah1, in particular the haloacid dehalogenase-like domain containing the DIDGT catalytic sequence, was resistant to the proteasomal degradation. The structural change of Pah1, as reflected by electrophoretic mobility shift, occurs through its phosphorylation by Pho85-Pho80, and the phosphorylation sites are located within its N- and C-terminal unfolded regions. Phosphorylation of Pah1 by Pho85-Pho80 inhibited its degradation, extending its half-life by ∼2-fold. The dephosphorylation of endogenously phosphorylated Pah1 by the Nem1-Spo7 protein phosphatase, which is highly specific for the sites phosphorylated by Pho85-Pho80, stimulated the 20S proteasomal degradation and reduced its half-life by 2.6-fold. These results indicate that the proteolysis of Pah1 by the 20S proteasome is controlled by its phosphorylation state.     

Development and in vitro evaluation of thiolated chitosan--Poly(methacrylic acid) nanoparticles as a local mucoadhesive delivery system

The main objective of this study was to develop a local, oral mucoadhesive metronidazole benzoate (MET) delivery system that can be applied and removed by the patient for the treatment of periodontal diseases. The results of present study revealed that the retention time of MET at its absorption site could be increased by formulating it into nanoparticles using thiolated chitosan (TCS)-poly(methacrylic acid) (PMAA). The nanoparticles of MET prepared from TCS-PMAA may represent a useful approach for targeting its release at its site of absorption, sustaining its release and improving its oral availability.     

The postcranial dimensions of the La Chapelle-aux-saints 1 Neandertal

The La Chapelle-aux-Saints 1 Neandertal has figured prominently in considerations of Neandertal body size and proportions. In this context, a reassessment of its major long bones and a reassembly of its principal pelvic elements (sacrum and right ilium) was undertaken. There are secure measurements for its humeral and radial lengths and its femoral head diameter, but the femoral and tibial lengths were almost certainly greater than previous values. The resultant humeral, femoral and tibial lengths are similar to those of other male Neandertals, its femoral head diameter is among the largest known for Middle and Late Pleistocene humans, but its radial length is relatively short. The pelvic assembly provides modest bi-iliac and inlet transverse diameters compared with the few sufficiently complete and undistorted Middle and Late Pleistocene archaic human pelves, but its dimensions are similar to those of large male early modern humans.     

The role of carboxyl groups on the chitosanase and CMCase activity of a bifunctional enzyme purified from a commercial cellulase with EDC modification

The carboxyl groups of the bifunctional cellulase–chitosanase (CCBE), purified from a commercial cellulase prepared from Trichoderma viride were modified using the water-soluble carbodiimide 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide (EDC). The EDC modified CCBE lost 80–90% of its chitosnase activity and 20% of its carboxylmethyl cellulase (CMCase) activity; meanwhile, its conformation changed slightly, which altered the substrate binding affinity to chitosan, without affecting its binding to CMC. However, the modification did not alter the structure integrity. The dynamic analysis of modification indicated that the CCBE possessed two carboxylates essential for its chitosanase activity and one carboxyl group for its CMCase activity. One of the two carboxylates involved in chitosanase activity was deduced to be the proton donator, and the other may function for substrate recognition, while the only catalytic carboxyl group for CMCase activity probably also acted as a proton donator.     

Subunit A of the E. coli ATP synthase: reconstitution and high resolution NMR with protein purified in a mixed polarity solvent

p23 is a regulatory co-chaperone of heat shock protein (Hsp) 90, but can also act as a general molecular chaperone by itself. Using novel point mutations of p23 that disrupt its interaction with Hsp90 we found its co-chaperone function to be required for its inhibitory effect on glucocorticoid receptor (GR). The C-terminal region of p23, which is required for its chaperone activity, is dispensable for inhibition of GR. Importantly, similar results were obtained with a constitutively active GR. Thus, the action of p23 on the nuclear stage of GR regulation requires its Hsp90 co-chaperone function, but not its chaperone activity.     

Detection of HBs antigen by hemagglutination inhibition tests; experiment with standardization]

The standarization of a reagent for automated detection of HBs antigen by an inhibition of the hemagglutination reaction is described. Its advantages are its specificity, its fast efficiency, its low cost. But, its handling is delicate and requires a good technicity.     

Comparative aspects of the area postrema: fine-structural considerations help to determine its function

The area postrema is a circumventricular organ of the fourth ventricle of the mammalian brain. Although there are distinct gross anatomical differences in the appearance of this organ between "lower" mammals such as rodents and lagomorphs and "higher" mammals such as carnivores and primates, its fine structure is remarkably similar in all species studied. There are many suggestions in the literature for a specific function for this area of the brain, ranging from its being a chemoreceptive trigger zone for the emetic response to its being a regulatory nucleus for the sleep cycle. The present report describes some comparative studies on the ultrastructure of this organ. This information is discussed in relation to what is known about the neurochemistry of the area postrema and its connections with other brain regions and visceral structures. A suggestion is offered that our current knowledge of the area postrema is consistent with its performing many of its proposed functions in the context of a regulatory ("fine-tuning") center for many autonomic functions.     

Activity of horse liver alcohol dehydrogenase SS with NADP (H) as coenzyme and its sensitivity to barbiturates

Alcohol dehydrogenase SS, free from other isoenzymes, has been purified from horse livers. The enzyme has high activity with NADP(H) as coenzyme. With NADPH its activity is 3 times more than with NADH. While its affinity for NADPH is less than for NADH, in comparison with the classical ADH its affinity for NADP(H) is increased. In its activity with NADP(H) and inhibition with barbiturates, ADH SS resembles aldehyde reductases.     

Apoptosis: final control point in cell biology

The discovery of apoptosis, a widespread and morphologically distinct form of physiological cell death, has had an extraordinary impact on cell biology. The importance of apoptosis stems from its active nature and its potential for controlling biological systems. The growing appreciation of the significance of this process has stimulated intense investigation into the molecular mechanisms involved and into its fundamental implications for developmental biology, immunology and oncology.     

Metabolism of phenols by Ochromonas danica

Abstract This study investigated the catabolic potential of a eukaryotic alga to degrade one of the most common organic pollutants, phenol. The alga, Ochromonas danica (993/28), was selected for study after screening for its heterotrophic capabilities. The catabolic versatility of the alga was elucidated by incubating with a variety of phenolic compounds. The alga removed phenol, all the cresol isomers and 3,4-xylenol from its incubation media, with phenol being removed more rapidly than any of its methylated homologues. Consequently, the alga was found to have a greater specificity for phenol than for o - or p -cresols. This study shows that O. danica could catabolize phenol and its methylated homologues.     

The periplasmic modifier protein for methanol dehydrogenase in the methylotrophs Methylophilus methylotrophus and Paracoccus denitrificans.

A modifier protein (M-protein), which increases the affinity of methanol dehydrogenase (MDH) for alcohols but decreases its affinity for formaldehyde, has been partially purified from Methylophilus methylotrophus and Paracoccus denitrificans. Analysis was complicated by non-protein factors in bacterial extracts that are able to mimic M-protein in one of its functions-that of increasing the activity of MDH with butane-1,3-diol in the dye-linked assay system. The 67 kDa polypeptide, previously identified as a subunit of the M-protein, is an unrelated cytoplasmic protein. The M-protein is exclusively periplasmic and is a multimeric protein with subunits of 45 kDa. The M-protein is active in the 'physiological' assay system with the specific cytochrome c electron acceptor for MDH, lowering its affinity for formaldehyde. It has its maximum effect when the ratio of M-protein:MDH is 1:5 but its concentration in the periplasm is much lower than 20% of that of MDH.     

Development and evaluation of a thermoreversible ovule formulation of stampidine,a novel nonspermicidal broad-spectrum anti-human immunodeficiency virus microbicide

Stampidine [2',3'-didehydro-2',3'-dideoxythymidine 5'-[p-bromophenyl methoxyalaninyl phosphate], a prodrug of stavudine (STV/d4T) with improved anti-HIV activity, is undergoing development as a novel nonspermicidal microbicide. Here, we report the stability of stampidine as a function of pH, preparation of a novel thermoreversible ovule formulation for mucosal delivery, its dissolution profile in synthetic vaginal fluid, and its mucosal toxicity potential as well as systemic absorption in the rabbit model. Stampidine was most stable under acidic conditions. Stampidine was solubilized in a thermoreversible ovule formulation composed of polyethylene glycol 400, polyethylene glycol fatty acid esters, and polysorbate 80. Does were exposed intravaginally for 14 days to an ovule formulation with and without 0.5%, 1%, or 2% stampidine corresponding to 1 x 107- to 4 x 107-fold higher than its in vitro anti-HIV IC50 value. Vaginal tissues harvested on Day 15 were evaluated for mucosal toxicity and cellular inflammation. Additionally, does were exposed intravaginally to stampidine, and plasma collected at various time points was assayed by analytical HPLC for the prodrug and its bioactive metabolites. Stampidine did not cause mucosal inflammation. The vaginal irritation scores for 0.5-2% stampidine were within the acceptable range for clinical trials. The prodrug and its major metabolites were undetectable in the blood plasma. The marked stability of stampidine at acidic pH, its rapid spreadability, together with its lack of mucosal toxicity or systemic absorption of stampidine via a thermoreversible ovule may provide the foundation for its clinical development as an easy-to-use, safe, and effective broad-spectrum anti-HIV microbicide without contraceptive activity.     

Stress-mediated adaptive response leading to genetic diversity and instability in metabolite contents of high medicinal value: an overview on Podophyllum hexandrum

Podophyllum hexandrum, known for its diversified clinical importance particularly for antineoplastic activity and valuable source for biological protection against high doses of radiation, has its unique position in the plant kingdom. Detailed understanding of mechanism and opportunity of chemical manipulations has amplified the scope of its bioactivity. Podophyllotoxin, the major active principle of this plant, has passed through various structural deviations with the basic aim of making the end product clinically more effective with minimal toxicity. However, over exploitation and limited growth has categorized this plant under endangered species. Depending upon the geographical variations, different species and subspecies of this plant have been explored. Morphological variations and quantitative differences in active principles are the major concern of its unstable medicinal value in whole and semifractionated preparations. The current review has addressed the issues related to the genetic diversity of P. hexandrum, extrinsic and intrinsic stresses responsible for its diversified nature, chemical modifications to enhance its multitasking bioactivity, and efforts for its cultivation and production of important metabolites to avoid collection of wild species due to its critically endangered nature.     

Cloning and expression in Escherichia coli of an extremely thermostable oligo-1,6-glucosidase gene from Bacillus thermoglucosidasius.

The gene for an extremely thermostable oligo-1,6-glucosidase (dextrin-6-alpha-D-glucanohydrolase; EC 3.2.1.10) of obligately thermophilic Bacillus thermoglucosidasius KP1006 was cloned within a 4.2-kilobase HindIII-PvuII fragment of DNA by using the plasmid pUC19 as a vector and Escherichia coli C600 as a host. The gene was transcribed, presumably from its own promoter, in E. coli. E. coli with the hybrid plasmid accumulated oligo-1,6-glucosidase mainly in the cytoplasm. The level of enzyme production was comparable to that observed for B. thermoglucosidasius. The enzyme coincided absolutely with the B. thermoglucosidasius enzyme in its molecular weight (60,000), in its electrophoretic behavior on denaturing and nondenaturing polyacrylamide gels, in the temperature dependency of its stability and activity, and in its antigenic determinants.     

Stabilization of glucoso-6-phosphate dehydrogenase by its substrate and cofactor in an ultrasonic field

The inactivation kinetics of glucoso-6-phosphate dehydrogenase (GPDH) and its complexes with glucoso-6-phosphate and NADP+ was characterized in aqueous solutions at 36-47 degrees C under treatment with low frequency (27 kHz, 60 W/cm2) and high frequency ultrasound (880 kHz, 1 W/cm2). To this end, we measured three effective first-order inactivation rate constants: thermal k(in)* , total (thermal and ultrasonic) kin, and ultrasonic kin (US). The values of the constants were found to be higher for the free enzyme than for its complexes GP-DH-GP and GPDH-NADP+ at all temperatures, which confirms the enzyme stabilization by its substrate and cofactor under both thermal and ultrasonic inactivation. Effective values of the activation energies (Ea) were determined and the preexponential factors of the rate constants and thermodynamic activation parameters of inactivation processes (deltaH*, deltaS*, and deltaG*) were calculated from the temperature dependences of the inactivation rate constants of GPDH and its complexes. The sonication of aqueous solutions of free GPDH and its complexes was accompanied by a reduction of Ea and deltaH* values in comparison with the corresponding values for thermal inactivation. The Ea, deltaH*, and deltaS* inactivation values for GPDH are lower than the corresponding values for its complexes. A linear dependence between the growth of the deltaH* and deltaS* values was observed for all the inactivation processes for free GPDH and its complexes.