ATP-binding domain of NTPase/helicase as a target for hepatitis C antiviral therapy

To enhance the inhibitory potential of 1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide (ribavirin) vs hepatitis C virus (HCV) NTPase/helicase, ribavirin-5'-triphosphate (ribavirin-TP) was synthesized and investigated. Ribavirin-TP was prepared with the use of modified Yoshikawa-Ludwig-Mishra-Broom procedure (cf. Mishra & Broom, 1991, J. Chem. Soc., Chem. Commun, 1276-1277) involving phosphorylation of unprotected nucleoside. Kinetic analysis revealed enhanced inhibitory potential of ribavirin-TP (IC50=40 microM) as compared to ribavirin (IC50 > 500 microM). Analysis of the inhibition type by means of graphical methods showed a competitive type of inhibition with respect to ATP. In view of the relatively low specificity towards nucleoside-5'-triphosphates (NTP) of the viral NTPase/helicases, it could not be ruled out that the investigated enzyme hydrolyzed the ribavirin-TP to less potent products. Investigations on non- hydrolysable analogs of ribavirin-TP or ribavirin-5'-diphosphate (ribavirin-DP) are currently under way.     

IpgD, a protein secreted by the type III secretion machinery of Shigella flexneri, is chaperoned by IpgE and implicated in entry focus formation

Invasion of epithelial cells by Shigella flexneri involves entry and intercellular dissemination. Entry of bacteria into non-phagocytic cells requires the IpaA-D proteins that are secreted by the Mxi-Spa type III secretion machinery. Type III secretion systems are found in several Gram-negative pathogens and serve to inject bacterial effector proteins directly into the cytoplasm of host cells. In this study, we have analysed the IpgD protein of S. flexneri, the gene of which is located on the virulence plasmid at the 5' end of the mxi-spa locus. We have shown that IpgD (i) is stored in the bacterial cytoplasm in association with a specific chaperone, IpgE; (ii) is secreted by the Mxi-Spa type III secretion system in amounts similar to those of the IpaA-D proteins; (iii) is associated with IpaA in the extracellular medium; and (iv) is involved in the modulation of the host cell response after contact of the bacterium with epithelial cells. This suggests that IpgD is an effector that might be injected into host cells to manipulate cellular processes during infection.     

甘肃盐池湾黑颈鹤亚成体夏季生境选择

开展对亚成体的研究,可以更加全面了解一个物种,进而更有效地开展保护工作。甘肃盐池湾国家级自然保护区是黑颈鹤（Grus nigricollis）成体的重要繁殖地和亚成体的重要栖息地之一。为研究甘肃盐池湾黑颈鹤亚成体生境选择,于2020年7月初至8月中旬在盐池湾党河湿地展开调查,并依据Johnson对生境选择空间尺度的划分,对亚成体活动区内各类型生境和觅食微生境的生境选择进行了研究。通过遥感影像解译和卫星跟踪分别获得各栖息地类型面积以及黑颈鹤的活动位点,利用核密度分析法估计活动区面积并利用Manly研究中的设计Ⅲ来研究活动区内各类型生境选择;通过选取利用样方和对照样方,使用χ2检验、独立样本t检验和Mann-Whitney U检验,对比检验样方数据,进行微生境选择的研究。结果表明,活动区内各类型生境中亚成体选择河流,拒绝戈壁和沼泽化草甸,对沼泽既不选择也不拒绝,而成体选择湖泊,没有利用河流,同时拒绝戈壁、山脉、沼泽化草甸和盐化草甸,对沼泽既不选择也不拒绝;觅食微生境选择中,亚成体选择平均植被盖度为57.07% ± 4.53%,基质类型为泥炭,基质硬度为中,主要植被黑褐苔草（Carex atrofusca）的微生境栖息,相比成体,亚成体选择的生境基质更硬,距道路距离更近,距房屋、河流、山脉和湖泊距离更远。亚成体的栖息地选择主要受到生境质量、生境资源有限性以及成体选择等因素的影响。在这些因素的影响下,亚成体与成体产生了生态位分离,并在栖息地选择上出现了分化。这种分化对亚成体的生存和成体的繁殖都有益,可以避免种内无效的冲突和竞争,有利于亚成体和成体的适合度增加。保护黑颈鹤的栖息环境需同时考虑到亚成体的选择和生存。     

Local and systemic effects of two herbivores with different feeding mechanisms on primary metabolism of cotton leaves

Caterpillars and spider mites are herbivores with different feeding mechanisms. Spider mites feed on the cell content via stylets, while caterpillars, as chewing herbivores, remove larger amounts of photosynthetically active tissue. We investigated local and systemic effects of short-term caterpillar and spider mite herbivory on cotton in terms of primary metabolism and growth processes. After short-term caterpillar feeding, leaf growth and water content were decreased in damaged leaves. The glutamate/glutamine ratio increased and other free amino acids were also affected. In contrast, mild spider mite infestation did not affect leaf growth or amino acid composition, but led to an increase in total nitrogen and sucrose concentrations. Both herbivores induced locally increased dark respiration, suggesting an increased mobilization of storage compounds potentially available for synthesis of defensive substances, but did not affect assimilation and transpiration. Systemically induced leaves were not significantly affected by the treatments performed in this study. The results show that cotton plants do not compensate the loss of photosynthetic tissue with higher photosynthetic efficiency of the remaining tissue. However, early plant responses to different herbivores leave their signature in primary metabolism, affecting leaf growth. Changes in amino acid concentrations, total nitrogen and sucrose content may affect subsequent herbivore performance.     

Cytologic observations in 35 individuals with a 5p- karyotype

Summary Chromosome investigation of 35 individuals with a 5p- karyotype and their families revealed the presence of 27 apparently terminal deletions, four interstitial deletions, and four translocations, including two familial cases. Four of the probands with simple deletions and one of the mothers were mosaics. Unusual chromosomal heteromorphism, as rendered visible after acridine orange staining, was observed on the short arm of chromosome 14 in two cases and, after heterochromatin staining, on chromosome 19 in one family. Measurement studies, carried out in probands with simple deletions and in two control groups, showed a short-arm loss clustering between 32% and 62% of the normal short-arm length. Using at least two complementary staining methods per proband, we found that the midportion of the 5p15 segment probably must be deleted to develop the typical clinical features of the cri du chat syndrome.     

Mismatched dNTP incorporation by DNA polymerase beta does not proceed via globally different conformational pathways

Understanding how DNA polymerases control fidelity requires elucidation of the mechanisms of matched and mismatched dNTP incorporations. Little is known about the latter because mismatched complexes do not crystallize readily. In this report, we employed small-angle X-ray scattering (SAXS) and structural modeling to probe the conformations of different intermediate states of mammalian DNA polymerase β (Pol β) in its wild-type and an error-prone variant, I260Q. Our structural results indicate that the mismatched ternary complex lies in-between the open and the closed forms, but more closely resembles the open form for WT and the closed form for I260Q. On the basis of molecular modeling, this over-stabilization of mismatched ternary complex of I260Q is likely caused by formation of a hydrogen bonding network between the side chains of Gln²⁶⁰, Tyr²⁹⁶, Glu²⁹⁵ and Arg²⁵⁸, freeing up Asp¹⁹² to coordinate MgdNTP. These results argue against recent reports suggesting that mismatched dNTP incorporations follow a conformational path distinctly different from that of matched dNTP incorporation, or that its conformational closing is a major contributor to fidelity.     

添加玉米和水稻秸秆对淹水土壤pH、二氧化碳及交换态铵的影响

通过温室土壤培养试验,研究了不同添加量玉米和水稻秸秆对淹水土壤pH、CO₂及交换态铵的影响.结果表明：在接近中性的土壤中加入秸秆可使土壤pH值降低,4 g·kg^-1玉米和水稻秸秆处理的土壤pH值均与对照差异显著（P＜0.05）,而1 g·kg^-1玉米和水稻秸秆处理与对照差异不显著（P＞0.05）.土壤溶液中的CO₂含量随秸秆添加量的增加而增大,1 g·kg^-1玉米和水稻秸秆处理的土壤溶液CO₂含量最大值分别为35.9％和31.9％（v/v）,与对照（25.8％）差异达显著水平（P＜0.05）,但两者间差异不显著（P＞0.05）;4 g·kg^-1玉米和水稻秸秆处理的土壤溶液CO₂含量最大值分别为54.2％和41.8％（v/v）,与对照差异极显著（P＜0.01）,两者间差异也达显著水平（P＜0.05）.在不施氮肥的情况下,添加秸秆可降低土壤铵态氮浓度,且铵态氮浓度随秸秆添加量的增加而降低,不同添加量处理间差异显著（P＜0.05）;在施入氮肥的情况下,1 g·kg^-1玉米和水稻秸秆处理提高了土壤铵态氮浓度,而4 g·kg^-1玉米和水稻秸秆处理降低了土壤铵态氮浓度.无论是否施入氮肥,玉米和水稻秸秆处理的土壤铵态氮浓度差异不显著（P＞0.05）.     

Reaction of laying hens to humans in the home or a novel environment

Tests assessing the fear of humans by measuring avoidance or approach reactions of the test animals towards humans can be useful tools in welfare assessment schemes. In this study, we wanted to compare tests assessing the reactions of laying hens towards humans in the home environment to a test performed in a novel environment. As well we investigated inter-test correlations in the home environment. We performed several tests to assess the hens’ approach and avoidance reactions towards an unfamiliar human inside the home environment of laying hens in 14 caged flocks and 10 free-range flocks. We measured the reactions of hens to a stationary person and a moving person approaching individual hens in both systems, as well as reactions to a stationary person trying to touch individual hens in non-cage systems and reactions of caged hens to a person passing by. Additionally, a standardised arena test was performed outside the home environment to compare the birds’ avoidance reactions towards an approaching human in a novel environment. In caged hens no significant correlations of tests performed inside the home environment with the arena test could be found, but there were moderate to high correlations (r_s > 0.6) with one parameter of the arena test in non-caged hens. The different tests applied inside the home environment correlated significantly and moderately to highly with each other, supporting the validity of these tests to measure fear of humans. The caged laying hens in our study reacted differently in the novel environment from the non-caged birds, indicating that the birds’ reactions towards humans in an unfamiliar environment depend on the housing system. In summary, a comparison of the level of fear of humans between flocks in different housing systems by using an arena test outside the home environment seems difficult and was not possible in the present study.     

Polymorphismus der Adenosindesaminase

Zusammenfassung Es wurden die Erythrocyten-ADA-Isoenzymmuster bei 500 unausgelesenen Personen der West-Berliner Bevölkerung untersucht. Die gefundenen Genhäufigkeiten betragen: ADA¹=0,942 und ADA²=0,058.

---

Adenosine deaminase polymorphismStudy on a population sample from West-Berlin

Summary The ADA isoenzyme patterns in the blood of 500 unselected individuals living in West-Berlin have been determined. The found gene frequencies are ADA¹=0.942 and ADA²=0.058.