Microplate-based screening for small molecule inhibitors of neuropilin-2/vascular endothelial growth factor-C interactions

Vascular endothelial growth factor-C (VEGF-C) is a secreted growth factor essential for lymphangiogenesis. VEGF-C functions in both physiological and pathological lymphangiogenesis, particularly in tumor metastasis, making it an attractive therapeutic target. Members of two families of cell surface receptors transduce VEGF-C signals: neuropilin-2 (Nrp2) and VEGF-receptor (VEGFR)-2/3. Nrp2 is a promising target for inhibition because it is highly expressed in lymphatic vessels. Here we describe a microplate-based assay for discovery of VEGF-C/Nrp2 inhibitors. We optimize this assay for use in screening an inhibitor library and identify three novel Nrp2/VEGF-C binding inhibitors from the National Institutes of Health (NIH) Clinical Collection small molecule library.     

Trapping Elusive Cats: Using Intensive Camera Trapping to Estimate the Density of a Rare African Felid

Camera trapping studies have become increasingly popular to produce population estimates of individually recognisable mammals. Yet, monitoring techniques for rare species which occur at extremely low densities are lacking. Additionally, species which have unpredictable movements may make obtaining reliable population estimates challenging due to low detectability. Our study explores the effectiveness of intensive camera trapping for estimating cheetah (Acinonyx jubatus) numbers. Using both a more traditional, systematic grid approach and pre-determined, targeted sites for camera placement, the cheetah population of the Northern Tuli Game Reserve, Botswana was sampled between December 2012 and October 2013. Placement of cameras in a regular grid pattern yielded very few (n = 9) cheetah images and these were insufficient to estimate cheetah density. However, pre-selected cheetah scent-marking posts provided 53 images of seven adult cheetahs (0.61 ± 0.18 cheetahs/100km²). While increasing the length of the camera trapping survey from 90 to 130 days increased the total number of cheetah images obtained (from 53 to 200), no new individuals were recorded and the estimated population density remained stable. Thus, our study demonstrates that targeted camera placement (irrespective of survey duration) is necessary for reliably assessing cheetah densities where populations are naturally very low or dominated by transient individuals. Significantly our approach can easily be applied to other rare predator species.     

Purification and characterization of soluble forms of human and rat stem cell factor recombinantly expressed by Escherichia coli and by Chinese hamster ovary cells.

Stem cell factor (SCF) is a novel, early-acting hematopoietic factor. It was isolated from the medium of a rat cell line in a soluble, processed form (Zsebo et al., 1990, Cell 63, 195). The cloned human and rat genes encode the soluble form plus additional C-terminal amino acids including a hydrophobic transmembrane domain (Martin et al., 1990, Cell 63, 203). We have recombinantly expressed forms of human and rat SCF corresponding to the soluble, processed form in Escherichia coli and in Chinese hamster ovary (CHO) cells. After expression in E. coli, folding and oxidation of the SCF polypeptides are required. The SCFs expressed in CHO cells are secreted into the medium in active state and, like the natural SCF, are glycosylated. Purification of the recombinant SCFs is described. Biological and biochemical characterization includes activity toward responsive human and mouse cell lines, N-terminal amino acid sequences, disulfide bond linkages, and sites of glycosylation.     

兰州唐古特白刺根际微生物分离鉴定

【目的】从兰州唐古特白刺根际分离得到对植物有潜在促生效果的功能微生物，为研发相关菌种制剂的研究奠定基础。【方法】通过平板划线法从其根际分离纯化出6株细菌，并对菌株进行形态特征观察、革兰氏染色等一系列生理生化试验。用藜麦检测各菌株的促生功能，并对具有优良促生作用的1个菌株16S rRNA基因进行分子鉴定及基因草图绘制。【结果】根据生化鉴定结果，6株细菌分别属于不动杆菌属(Acinetobacter)、土壤杆菌属(Agrobacterium)、类芽孢杆菌属(Paenibacillus)、芽孢杆菌属(Bacillus)、鞘氨醇单胞菌属(Sphingomonas)和假单胞菌属(Pseudomonas)。其中，16S rRNA基因鉴定BC4属于肠杆菌属(Enterobacter)，具有较好的促生效果。【结论】BC4具有较好的促生效果，为兰州唐古特白刺菌种资源的开发和利用提供了一定的理论依据。     

阿拉善高原2种荒漠植物根系构型及生态适应性特征

根系构型决定了植物对资源的吸收方式，根系构型的变化是植物对环境的生态适应和有效生存策略。在阿拉善高原西南缘红砂（Reaumuria soongarica）-珍珠猪毛菜（Salsola passerina）混生群落采用传统挖掘法收集两种植物根系，基于量化的根系形态指标，利用几何拓扑学及分形理论分析了根系构型特征，探讨了该地区2种植物对干旱生境的生态适应策略。结果表明：红砂和珍珠猪毛菜根系均以水平分布占优，根系浅层化分布明显，混生的两种植物占据不同的生态位；2种荒漠植物均具有较大的比根长（SRL）和比表面积（SRA），红砂SRL=21.3 cm/g，SRA=7.6 cm²/g，珍珠SRL=22.4 cm/g，SRA=6.5 cm²/g，有利于水分和养分的获取；红砂根系拓扑指数（TI）、修正拓扑参数（q_a和q_b）分别为0.86、0.52、0.49，珍珠猪毛菜对应参数分别为0.93、0.76、0.73，表明2种植物根系均趋向于鱼尾形分支结构；根系分形维数值（FD_红=1.488、FD_珍=1.422）较小，而分形丰度值（lgK_红=1.855、lgK_珍=1.774）较大，表明2种植物分支相对简单，但空间拓展能力强，有利于对营养空间的占有。上述特征可能是阿拉善西南缘红砂-珍珠猪毛菜群落2种荒漠植物植物对干旱贫瘠生境的重要生态适应策略。     

Fluctuating asymmetry of morphometric characters in house mice: the effects of age, sex, and phenotypical extremeness in a randombred population

Fluctuating asymmetry (FA) was calculated for nine paired morphometric characters in a randombred population of house mice in order to assess the effects of age, sex, and phenotypical extremeness. FA did not significantly vary between sexes for any of the characters, and age proved to be significant only for innominate length. Additionally, FA values in high extreme individuals and low extreme individuals were compared to those for intermediate individuals to discover whether extreme individuals showed increased fluctuating asymmetry (decreased developmental stability) due to increased levels of homozygosity. Differences in fluctuating asymmetry magnitudes were found between the groups, although the extreme individuals did not consistently show increased fluctuating asymmetry and FA differences also showed no significant association with the level of heritability of each character. Potential reasons for these findings are discussed.     

The ultrastructure of mouse testicular interstitial tissue containing plutonium-239 and its significance in explaining the observed distribution of plutonium in the testis.

The technique of autoradiography with Araldite-embedded sections was used to study the distribution of 239Pu in mouse testis at various times post-injection. Adjacent sections were examined with both the light microscope and electron microscope. The autoradiographs showed that from 1 week to 3 months postinjection, most 239Pu in located in interstitial tissue. The major change in distribution observed was that the early diffuse deposit in interstitial tissue is concentrated in macrophages with increasing time post-injection. This is a real change of distribution as the amount of 239Pu in mouse testis remains constant from 1 week to 3 months post-injection. Study of the ultrastructure of interstitial tissue indicated that the accumulation of 239Pu in macrophages may be brought about in two ways. First, there may be phagocytosis of dead cells containing 239Pu. Second, 239Pu may follow the transfer of waste products of hormone synthesis from Leydig cells into macrophages. The significance of these observations is discussed with regard to the deposition of 239Pu in human testis.     

Effect of Cimetidine and other antihistaminics on the elimination of aminopyrine,phenacetin and caffein

Cimetidine is widely prescribed for the treatment of peptic ulcer disease and has recently been shown to inhibit the metabolism of warfarin, antipyrine and diazepam. To further examine this phenomenon we investigated the effect of various doses of cimetidine and other related drugs on ¹⁴C-aminopyrine, ¹⁴C-phenacetin and ¹⁴C-caffeine breath tests. Cimetidine caused a dose-related inhibition of the metabolism of aminopyrine and caffeine but had no effect on the phenacetin breath test. Metiamide, H₁-antihistamines, phenothiazines and local anesthetics also inhibited the aminopyrine breath test. Cyproheptadine had no effect on either phenacetin or caffeine elimination. This study demonstrates a complex drug-drug interaction which may have widespread clinical implications.