Participation of the conjugated diene part for potent cytotoxicity of callystatin A, a spongean polyketide

5-epi, 10-epi, 8-Deethyl, and 10-demethyl analogues of callystatin A, a potent cytotoxic spongean polyketide, were synthesized to elucidate structure-requirement for cytotoxic potency. Inversion of the asymmetric center at C-10 in callystatin A minimally affected the activity, while lack of the 10-methyl group in callystatin A decreased cytotoxicity. In addition, the C-5 epimer and the 8-deethyl analogue of callystatin A showed weaker cytotoxicity.     

Leptostatin: a synthetic hybrid of the cytotoxic polyketides callystatin A and leptomycin B

Four stereoisomeric hybrids of the polyketide natural products callystatin A and leptomycin B have been prepared by parallel synthetic routes involving chiral allenylstannane methodology. Like their natural counterparts, these hybrids exhibit nanomolar levels of cytotoxicity toward HCT-116 human colon cancer cells.     

Design and synthesis of a selective EP4-receptor agonist. Part 3: 16-phenyl-5-thiaPGE(1) and 9-beta-halo derivatives with improved stability

To identify a new selective EP4-agonist with improved chemical stability, further chemical modification of those reported previously was continued. We focused our attention on chemical modification of the alpha chain of 3,7-dithiaPGE(1) and selected 5-thiaPGE(1) as a new chemical lead. Introduction of an optimized omega chain to the 5-thiaPG skeleton afforded m-methoxymethyl derivative 33a, which showed the most potent EP4-receptor agonist activity and good subtype-selectivity both in vitro and in vivo. 9beta-HaloPGF derivatives were also synthesized and biologically evaluated in an attempt to block self-degradation of the beta-hydroxyketone moiety. Among these series, and 39b showed potent agonist activity and good subtype-selectivity. Structure-activity relationships (SARs) are also discussed.     

Multi parameter in vitro testing of ratjadone using flow cytometry

Ratjadone, isolated from the myxobacterium Sorangium cellulosum, belongs to the family of so-called orphan ligands, which includes leptomycin, callystatin and other compounds. In previous screening tests, ratjadone revealed a growth inhibitory effect against bacteria, yeast and human cancer cells. Following these first results, ratjadone was tested on several human tumour cell lines (Jurkat, HepG2, U87-MG) and, as a control, on a non-tumour cell line (RLC18) for its mode of action. The cell analysis was carried out by flow cytometry. This comprised cell density measurements, live-dead analysis, cell-cycle analysis and detection of apoptosis. First experiments confirmed the growth inhibitory effect on any chosen tumour cell line. Following these results a dose effect relationship was monitored, confirming the high effectiveness of ratjadone against cell growth at nanomolar concentration. Cell cycle analysis has shown that ratjadone intervenes in the cell cycle by arresting the cells in G1-phase. Biological testing of additional ratjadone derivatives with changed configuration and stereochemistry, identified the pharmacophoric site of the molecule.     

Differential effects of long-term hypoxia on norepinephrine turnover in brain stem cell groups.

The influence of long-term hypoxia on noradrenergic cell groups in the brain stem was assessed by estimating the changes in norepinephrine (NE) turnover in A1, A2 (subdivided into anterior and posterior parts), A5, and A6 groups in rats exposed to hypoxia (10% O2-90% N2) for 14 days. The NE turnover was decreased in A5 and A6 groups but failed to change significantly in A1. The NE turnover was increased in the posterior part of A2 and remained unaltered in the anterior part. In normoxic rats, the hypotensive drug dihydralazine induced a reverse effect, namely increased NE turnover in anterior A2 and no change in posterior A2. The neurochemical responses to hypoxia were abolished by transection of carotid sinus nerves. The results show that long-term hypoxia exerts differential effects on the noradrenergic cell groups located in the brain stem. Peripheral chemosensory inputs control the hypoxia-induced noradrenergic alterations. The A2 cell group displays a functional subdivision: the posterior part is influenced by peripheral chemosensory inputs, whereas the anterior part may be concerned with barosensitivity.     

Interaction of arylpiperazine ligands with the hydrophobic part of the 5-HT1A receptor binding site

A flexible docking of a series of arylpiperazine derivatives with structurally different aryl part to the binding site of a model of human 5-HT1A receptor was exercised. The influence of structure and hydrophobic properties of aryl moiety on binding affinities was discussed and a model for ligand binding in the hydrophobic part of the binding site was proposed.     

根区水肥空间耦合对冬小麦生长及产量的影响

利用管栽试验研究了根区不同湿润方式（整体湿润、上湿下干、上干下湿）、施肥方式（整体施肥、上层施肥、下层施肥）及其耦合对冬小麦不同生育期生长及产量的影响.结果表明：下层施肥方式显著降低了分蘖期冬小麦的株高和叶面积,而不同湿润方式对分蘖期株高和叶面积的影响不显著,拔节期水肥同区方式的株高大于水肥异区方式,表现出协同耦合效应.上干下湿方式和下层施肥方式显著降低了根系干物质量、地上部干物质量和总干物质量,上层施肥方式有利于增加冬小麦生物量,而上湿下干方式与施肥处理对地上部干物质量和总干物质量的耦合效应明显.水肥同区处理的根冠比高于水肥异区处理;上干下湿方式的水分利用效率显著高于整体湿润和上湿下干方式,水肥同区处理的水分利用效率高于水肥异区处理,但下层施肥方式的水分利用效率较低.与上干下湿方式相比,上湿下干和整体湿润方式的冬小麦单穗粒数分别增加了41.7%和61.9%,上层施肥和整体施肥方式的单穗粒数高于下层施肥方式,上湿下干方式与施肥处理对小麦产量及产量构成因素（除千粒重外）的水肥耦合效应明显.不同水肥处理主要通过影响单穗粒数来影响冬小麦产量.     

Heterologous expression of functionally active enterolysin A, class III bacteriocin from Enterococcus faecalis, in Escherichia coli

The heterologous expression of enterolysin A (EnlA), heat-labile class III bacteriocin from Enterococcus faecalis II/1 with anti-listerial activity, was studied in Escherichia coli. The PCR amplified products of enterolysin A structural gene, N-terminal part of EnlA with endopeptidase-like activity and C-terminal part of EnlA similar to a lysis gene of bacteriophage, were cloned in prelinearized pQE-30UA expression vector. The expression of EnlA structural gene led to the synthesis and secretion of functional-active His-tagged enterolysin A protein, which was purified to homogeneity using His-Select™ Cartridge and was shown to be fully active against the indicator strain. The expression of N-terminal or C-terminal part of EnlA and deletion of last 58 amino acids from C-terminal domain of EnlA led to the synthesis of biologically non-active proteins.     

Studies on the Characteristics of K,Na Content in Aneurolepidium chinense Grassland of Northeast China

The characteristics of K, Na contents of the pasture-for-cutting of Aneurolepidiurn chinense grassland of Northeast China were studied. Results showed that: K concentration of every organ of the aboveground part of A. ch inense was highest in early growing season, then decreased gradually as plant continues to grow. The variation of Na concentration was more or less similar to that of K, but there was a sudden increase in August. The concentrations of K, Na in the aboveground part of the community were higher than those in the underground part..The variations of K, Na accumulations of the aboveground part of ,A. chinense in the growing season were of the single-peak type, the peaks of K and Na appeared in July and August respectively. The variations of K, Na accumulations of the aboveground parts of Carex duriuscula and Heleocharis acicularis were similar to those of the biomass, which were of the double-peak type. The accumulations of K and Na in the underground part of community were significantly higher than those in the aboveground part; the accumulations of K, Na in leaf and stem were very close to each other. The K, Na accumulations in the community were accounted for 0.25% and 0.71% of the total reserves of the soil, 2.31% and 0.93% of the soluble plus exchangeable reserves of the soil respectively.     

Thymidylate synthetase in the antimesometrial and mesometrial portions of decidual tissue under normal conditions and following application of the antifolate preparation chloridine]

Specific activity of thymidylate synthetase (TMS) under normal conditions and after the action of pyrimethamine was investigated in antimesometrial (A) and mesometrial (M) parts of rat decidua. On the 9th-11th days of pregnancy specific activity of TMS in the A part proved to be higher than in the M part of decidua, and the former one reached its maximum by the 10th day. Specific activity of TMS in the M part decreased gradually from the 9th to 11th days of gestation. Pyrimethamine, applied on the 9th day of pregnancy caused a wave-like increase in the specific activity of TMS in the A part; this elevation was not so pronounced in the M part; A study of the physico-chemical properties of TMS from the decidua and the embryos of rats showed that the enzyme had a molecular weight of 58000, an optimal pH of 6.9, and could be quickly inactivated by heating.     

Effects of leachates of the invasive plant, Ageratina adenophora (Sprengel) on soil microbial community

The invasive plant Ageratina adenophora (Sprengel) changed soil microbial communities in the invaded area to facilitate its growth and inhibit native plants. However, little is known about the driving forces underlying the alteration of soil biota. Leachates from root and aerial part (stem and leaves) of A. adenophora were mixed into soil to imitate field invasion processes for evaluation of its impact on invasion of soil microbial community. The results indicated that soil microbial community was significantly changed when the soil taken from the newly-invaded area was treated with A. adenophora root and aerial part leachates for 3 and 5 weeks, respectively. The biota of newly invaded soil treated with concentration of 100 mg/mL A.adenophora leachates was much closer to that of heavily invaded soil, but was significantly different from that of control soil (newly invaded soil without treatment). A.adenophora leachates promoted growth of the seven dominant rhizosphere bacterial species in the invaded soil. The effect of A.adenophora leachates on soil biota and dominant rhizosphere bacteria was positively correlated with the concentration of leachates, however, the effect of root leachates was stronger than the aerial part leachates. It is assumed that A.adenophora change soil microbial community via nutritional and chemical communication, which helps it in better colonization of the invaded soil.     

Effects of leachates of the invasive plant, Ageratina adenophora (Sprengel) on soil microbial community

The invasive plant Ageratina adenophora (Sprengel) changed soil microbial communities in the invaded area to facilitate its growth and inhibit native plants. However, little is known about the driving forces underlying the alteration of soil biota. Leachates from root and aerial part (stem and leaves) of A. adenophora were mixed into soil to imitate field invasion processes for evaluation of its impact on invasion of soil microbial community. The results indicated that soil microbial community was significantly changed when the soil taken from the newly-invaded area was treated with A. adenophora root and aerial part leachates for 3 and 5 weeks, respectively. The biota of newly invaded soil treated with concentration of 100 mg/mL A.adenophora leachates was much closer to that of heavily invaded soil, but was significantly different from that of control soil (newly invaded soil without treatment). A.adenophora leachates promoted growth of the seven dominant rhizosphere bacterial species in the invaded soil. The effect of A.adenophora leachates on soil biota and dominant rhizosphere bacteria was positively correlated with the concentration of leachates, however, the effect of root leachates was stronger than the aerial part leachates. It is assumed that A.adenophora change soil microbial community via nutritional and chemical communication, which helps it in better colonization of the invaded soil.     

苹果蠹蛾老熟幼虫诱杀技术

本文对简易杀虫诱集带诱杀苹果蠹蛾Cydia pomonella(L.)老熟幼虫的效果进行了研究。发现在编织袋,旧衣物,瓦楞纸3种材料中,旧衣物对苹果蠹蛾的老熟幼虫捕获效果最好,其诱捕量占总诱捕量的68.42%;在诱集带中添加的农药中(包括乐斯本、敌敌畏、白僵菌、灭幼脲3号、高效氯氰菊酯等),乐斯本的杀虫效果最好,幼虫死亡率达到98.20%;在不同浓度的乐斯本溶液中,500倍乐斯本杀虫效果最好,幼虫死亡率达到91.61%,且对幼虫没有驱避作用;500倍乐斯本+50倍白僵菌药杀虫诱集带有效期长达40多天,37d后对幼虫的杀伤效果依然达到96%以上,作为老熟幼虫的防治措施十分理想。     

Development of a new doubly-labeled fluorescent ceramide probe for monitoring the metabolism of sphingolipids in living cells

A new ceramide analog, 1, containing two fluorescent dyes, NBD in the N-acyl part and KFL5 in the alkyl part, was synthesized. The fluorescence from both NBD and KFL5 was detected in living cells in a time-dependent manner. A multi-wavelength fluorescence detector was used to detect ceramide metabolites including sphingosine, sphingosine-1-phosphate, glucosylceramide, and sphingomyelin, which are connected to the fluorescent dyes, simultaneously in a single TLC plate.     

Creation of maize lines inducing maternal haploids with high frequency

The new inducers of maternal haploids have been created. The results on their inducing capacity in crossing with MK01u and A619 lines and MK01u x A619 hybrid are presented. Pollination with pollen of the new inducers allowed to obtain in average 6.5% of haploids among the three maternal genotypes. The ZMS and KMS initial lines induced 2.3% and 2.0% of haploids respectively. The frequency of haploids was studied in different parts of ears. It was found that in the higher part of an ear where the egg cells are the youngest during flowering the frequency of haploids was almost three times higher than in the lower part of an ear and more than twice higher than in the middle part of an ear. It was assumed that early pollination may increase frequency of obtaining haploids.     

Distribution of acetylcholinesterase and monoamine oxidase in the amygdala of the guinea pig

Summary A study of the amygdala of the guinea pig was carried out on material stained by the Nissl, acetylcholinesterase (AChE) and monoamine oxidase (MAO) methods. The material stained for Nissl substance was used primarily as a reference in determining the distribution of the two enzymes. Regional differences in cell size and/or distribution were noted within the lateral, basal, medial and cortical nuclei. In the AChE preparations, it was observed that the large-celled part of the basal nucleus stained very intensely, the small-celled part of the basal nucleus and ventromedial part of the lateral nucleus more moderately, and the dorsolateral part of the lateral nucleus and cortical nucleus lightly. The central and medial nuclei showed almost no reaction. With the MAO method, the greatest staining reaction was seen in the medial nucleus, the medial part of the cortical nucleus, the anterior amygdaloid area and the ventromedial wedge of the putamen adjacent to the central nucleus. In addition, fibres of the stria terminalis stained very darkly.These findings are discussed in relation to the observations of previous authors employing the same methods.Supported in part by the Canadian Medical Research Council Grant No. M.T. 870 and U.S. Public Health Service Grant No. NS-07998. This aid is gratefully acknowledged. We are indebted to Dr. Gorm Danscher for additional material and to Mr. A. Meier, Mrs. L. Munkøe, Mrs. K. Sørensen, Miss M. Sørensen, Miss D. Valgaard, and Miss B. Ørum for skillful assistance.     

Foot-and-mouth disease virus 2A protease mediates cleavage in attenuated Sabin 3 poliovirus vectors engineered for delivery of foreign antigens.

Poliovirus vectors are being studied as potential vaccine delivery systems, with foreign genetic sequences incorporated as part of the viral genome. The foreign sequences are expressed as part of the viral polyprotein. Addition of proteolytic cleavage sites at the junction of the foreign polypeptide and the viral proteins results in cleavage during polyprotein processing. The ability of foot-and-mouth disease virus (FMDV) 2A to mediate proteolytic cleavage in the context of poliovirus vectors was studied. The results demonstrate that FMDV 2A is able to generate cleavage of the foreign antigen from the viral polyprotein. A second cleavage event between the FMDV 2A peptide and the foreign protein was also observed.     

Genetic variation in vivo and proposed functional domains of the 5'' noncoding region of poliovirus RNA.

Poliovirus has a single-stranded RNA genome of about 7,440 nucleotides (nt) with an unusually long 750-nt noncoding region in the 5' end (5'NCR). Several regulatory functions have been assigned to the 5'NCR. We sequenced the 5'NCRs of 33 wild-type 3 poliovirus strains to study the range and distribution of naturally occurring sequence variations. In this regard, the 5'NCR can be divided into a conserved part (nt 1 to 650) and a hypervariable part (nt 651 to 750). In the conserved part, altogether 234 unevenly distributed nucleotide positions (36%) showed variation. When these positions were plotted against the predicted secondary-structure models, it was found that the existence of most of the proposed stem-loop structures was supported by extensive structure-conserving substitutions in the stems. Regions with conserved sequences, as well as mutational hot spots, were observed. The hypervariable part of the 5'NCR varied up to 56% between the strains studied. The A + U percentage was significantly higher than in the conserved part. The number of AUG codons varied between 5 and 15 in the conserved part of the 5'NCR, while none was found in the hypervariable part. These results provide information that can be used in site-directed mutagenesis and other approaches targeted to reveal the functional domains of the 5'NCR.     

Histological and three dimensional organization of the odontogenic organ in the lower incisor of 100 gram rats.

A three dimensional reconstruction of the epithelial tissue at the apical end of the lower rat incisor was made from serial 1 mum thick cross sections. This tissue formed an elongated structure, called the odontogenic organ, which was composed of a bulbous and a "U"-shaped part. Both parts were joined to one another at the posterior aspect of the apical foramen. The bulbous part of the odontogenic organ was situated at the lingual side of the "U"-shaped part and protruded anteriorly over the pulp. It was formed by cells of the outer dental epithelium and stellate reticulum whose organization suggested that the bulbous part was important in the production of cells for renewal of all the epithelia of the incisor. The "U"-shaped part of the odontogenic organ was apparently derived from the bulbous part and delineated the pulp by forming the lateral, mesial and labial sidewalls around the apical foramen. It was composed of all the epithelial cell types recognizable as precursors to (a) cells of the enamel organ which form the enamel, and (b) Hertwig's epithelial root sheath, a part of the odontogenic organ which induces the formation of dentin on the lingual aspect of the incisor.