Identification of the 5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine derivatives as highly selective PDE4B inhibitors

A PDE4B subtype selective inhibitor is expected to have a wider therapeutic window than non-selective PDE4 inhibitors. In this Letter, two series of 7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine derivatives and 5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine derivatives were evaluated for their PDE4B subtype selectivity using human PDE4B2 and PDE4D2 full length enzymes. To improve their PDE4B selectivity over PDE4D, we optimized the substituents on the pyrimidine ring and the side chain phenyl ring, resulting in several derivatives with more than 100-fold selectivity for PDE4B. Consequently, we identified 2-(3-chloro-4-methoxy-phenyl)-5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine derivative 54 as a highly selective PDE4B inhibitor, which had potent hPDE4B inhibitory activity with an IC₅₀ value of 3.0 nM and 433-fold PDE4B selectivity over PDE4D.     

Development of potent antagonists for formyl peptide receptor 1 based on Boc-Phe-d-Leu-Phe-d-Leu-Phe-OH

While stimulation of formyl peptide receptors (FPRs) on the surface of human neutrophils induces several immune responses, under conditions of continuous activation of the receptor by agonists such as formyl-Met-Leu-Phe-OH (fMLP), neutrophil-dependent tissue damage ensues. Thus, FPR antagonists could be anticipated as drugs for FPR-related disease. In this study, Boc-Phe-d-Leu-Phe-d-Leu-Phe-OH (Boc-FlFlF), one of several FPR subtype selective antagonists, was chosen and the positions at the Phe residues were optimized. We found that substitution with unnatural amino acids resulted in an improvement of two orders of magnitude. The most potent antagonist indicated FPR subtype selectivity at 1 μM. In addition to finding a potent antagonist, the structure–activity trends observed in this study should be valuable in designing a new type of FPR subtype selective antagonist.     

Gestational diabetes mellitus (GDM) decreases butyrylcholinesterase (BChE) activity and changes its relationship with lipids

Many conditions interfere with butyrylcholinesterase (BChE) activity, e.g., pregnancy or presence of the BCHE gene variant −116A can decrease activity whereas obesity and types I and II diabetes mellitus can increase activity. In this study, we examined BChE activity, −116A and 1615A BCHE gene variants, and anthropometric and biochemical variables associated with diabetes in patients with gestational diabetes mellitus (GDM) and in healthy pregnant women. BChE activity was measured spectrophotometrically using propionylthiocholine as substrate and genotyping of the −116 and 1615 sites of the BCHE gene was done with a TaqMan SNP genotyping assay. Three groups were studied: 150 patients with GDM, 295 healthy pregnant women and 156 non-pregnant healthy women. Mean BChE activity was significantly lower in healthy pregnant women than in women from the general population and was further reduced in GDM patients. BChE activity was significantly reduced in carriers of −116A in GDM patients and healthy pregnant women. Although GDM patients had a significantly higher mean body mass index (BMI) and triglycerides than healthy pregnant women, they had lower mean BChE activity, suggesting that the lowering effect of GDM on BChE activity was stronger than the characteristic enhancing effect of increased BMI and triglycerides.     

Don’t miss the forest for the trees! Evidence for vertical differences in the response of plant diversity to disturbance in a tropical rain forest

Ecological studies in tropical rain forests traditionally focus on trees above a threshold diameter at breast height (dbh), since ignoring plant species of the other structural compartments is believed to be an acceptable tradeoff between exhaustiveness and effectiveness. However, the consequences of missing species below a threshold dbh value have been largely neglected so far. We evaluated whether the response of species diversity of ≥10-cm dbh trees was similar to the response of other structural ensembles (namely treelets, saplings, and terricolous herbs) in a lowland tropical rain forest, to three disturbance regimes: natural gap dynamics (control), and selective logging with and without additional thinning. We studied forest vegetation composition and diversity in a 20-yr replicated field experiment comprising nine 1 ha permanent plots established in a semi-deciduous rain forest of the Congo Basin and equally distributed among the three treatments. Once corrected by stem density, species richness was similar between logged (20 years since logging) and untouched old-growth forest stands with respect of trees, but higher with respect of treelets. As disturbance intensity increased, species richness increased within sapling layers but decreased within herb layers, while species spatial turnover (beta diversity) increased in both cases. Regarding the parameters of the partitioned rarefaction curves and relative abundance distribution curves, no correlation was found between trees and any of the other structural compartments. Whilst tree and treelet species composition was similar among treatments, the understories still reflected past disturbance intensity, with a strong response of the sapling and herb layers. These results show that ecological studies based solely on tree layers (dbh ≥ 10 cm) are misleading because their response to disturbance cannot be used as a surrogate for the response of other structural ensembles. Long-lasting effects of anthropogenic disturbance on the sapling bank and the herb layer may durably influence the long-term forest dynamics. Since overstory but not understory plant communities have recovered from human disturbances 20 years after silvicultural operations, African tropical rain forest ecosystems may not be as resilient to selective logging as previously thought.     

Selective adsorption and recovery of precious metal ions using protein-rich biomass as efficient adsorbents

Various types of protein-rich biomass were examined as selective and environment-friendly adsorbents for precious metal ions. In the presence of base metal ions, Au³⁺, Pd²⁺ and Pt⁴⁺ ions were selectively adsorbed to samples of protein-rich biomass. Among the biomass samples tested, egg-shell membrane exhibited the highest adsorption ability and had high selectivity for Au, Pd and Pt ions. The maximum adsorption amount of Au, Pd and Pt ions to egg-shell membrane was approximately 250, 110 and 50 mg/g, respectively, in the presence of 0.1 M HCl. Microscopic observations and metal-ion desorption studies suggested that the precious metal ions were adsorbed and a portion of them was reduced to form metal nanoparticles on the egg-shell membrane, leading to high adsorption ratios. Investigations using glycoproteins indicated the importance of sugar chains in the adsorption of Au ions to the egg-shell membrane. Successful recovery of Au, Pd and Pt ions from industrial waste solutions was also demonstrated using egg-shell membrane. Biomass sheets (1 mm thick) made from egg-shell membrane also exhibited adsorption abilities for precious metal ions.     

Evaluation of spice and herb as phyto-derived selective modulators of human retinaldehyde dehydrogenases using a simple in vitro method

Selective modulation of retinaldehyde dehydrogenases (RALDHs)—the main aldehyde dehydrogenase (ALDH) enzymes converting retinal into retinoic acid (RA), is very important not only in the RA signaling pathway but also for the potential regulatory effects on RALDH isozyme-specific processes and RALDH-related cancers. However, very few selective modulators for RALDHs have been identified, partly due to variable overexpression protocols of RALDHs and insensitive activity assay that needs to be addressed. In the present study, deletion of the N-terminal disordered regions is found to enable simple preparation of all RALDHs and their closest paralog ALDH2 using a single protocol. Fluorescence-based activity assay was employed for enzymatic activity investigation and screening for RALDH-specific modulators from extracts of various spices and herbs that are well-known for containing many phyto-derived anti-cancer constituents. Under the established conditions, spice and herb extracts exhibited differential regulatory effects on RALDHs/ALDH2 with several extracts showing potential selective inhibition of the activity of RALDHs. In addition, the presence of magnesium ions was shown to significantly increase the activity for the natural substrate retinal of RALDH3 but not the others, while His-tag cleavage considerably increased the activity of ALDH2 for the non-specific substrate retinal. Altogether we propose a readily reproducible workflow to find selective modulators for RALDHs and suggest potential sources of selective modulators from spices and herbs.     

微卫星标记的制备策略

微卫星标记作为一类重要的、成熟的分子遗传标记,凭借自身的高变异性、孟德尔遗传模 式、共显性遗传方式等优点,在遗传及其它相关领域发挥了十分重要的作用,而其主要的缺点是 必须知道所研究生物的微卫星序列及其旁侧序列。随着人类及模式生物基因组的深入研究,微 卫星标记的制备经历了从费时、费力、低效的传统法到周期短、效率高的富集法(选择性杂交法和 FIASCO法);从基因组构建文库筛选到公共数据库ESTs发掘;从实验室自制到花钱购买。总之, 随着方法与技术的不断改进,从目标生物中获得微卫星标记变得越来越简单、经济。     

一种从土壤样品中选择性分离假诺卡氏菌的方法

为提高假诺卡氏放线菌的分离效率,根据其营养特性和对抗生素的敏感性,设计、检验了5种选择性分离培养基;实验检测了模式菌株在不同培养基上的生长情况,结果表明培养基S1和S2对假诺卡氏菌的生长有显著的选择性。经该方法从韩国、印度尼西亚和中国广西地区不同土样中分离到一些假诺卡氏放线菌。     

低心血管疾病风险选择性环氧合酶-2抑制剂的虚拟筛选

摘要 目的：寻找具有血栓素A2受体（Thromboxane A2 receptor,TP）抑制作用的选择性环氧合酶-2（Cyclooxygenase-2,COX-2）抑制剂,以期降低其心血管疾病风险。方法：本研究从公开数据库中获取了512种TP抑制剂,通过分子对接、分子动力学模拟和ADMET预测,筛选出化合物TP84。结果：分子对接结果显示,与先前获批的选择性COX-2抑制剂罗非昔布相比,TP84对COX-2的亲和力更高,对环氧合酶-1（Cyclooxygenase-1,COX-1）的亲和力更低;分子动力学模拟进一步表明,模拟过程中TP84与COX-1的结合不稳定,而TP84能稳定结合COX-2,与COX-2的结合自由能是COX-1的3倍;此外,根据ADMET预测,TP84的药物化学、吸收、分布、代谢、排泄和毒性处于类药物候选物的可接受范围内。结论：TP84是一种潜在的低心血管疾病风险选择性COX-2抑制剂。     

Selective genotyping to detect quantitative trait loci affecting multiple traits: interval mapping analysis

 Segregating quantitative trait loci can be detected via linkage to genetic markers. By selectively genotyping individuals with extreme phenotypes for the quantitative trait, the power per individual genotyped is increased at the expense of the power per individual phenotyped, but linear-model estimates of the quantitative-locus effect will be biased. The properties of single- and multiple-trait maximum-likelihood estimates of quantitative-loci parameters derived from selectively genotyped samples were investigated using Monte-Carlo simulations of backcross populations. All individuals with trait records were included in the analyses. All quantitative-locus parameters and the residual correlation were unbiasedly estimated by multiple-trait maximum-likelihood methodology. With single-trait maximum-likelihood, unbiased estimates for quantitative-locus effect and location, and the residual variance, were obtained for the trait under selection, but biased estimates were derived for a correlated trait that was analyzed separately. When an effect of the QTL was simulated only on the trait under selection, a “ghost” effect was also found for the correlated trait. Furthermore, if an effect was simulated only for the correlated trait, then the statistical power was less than that obtained with a random sample of equal size. With multiple-trait analyses, the power of quantitative-trait locus detection was always greater with selective genotyping. Received: 23 February 1998 / Accepted: 15 May 1998