Mutational analysis of slyD, an Escherichia coli gene encoding a protein of the FKBP immunophilin family

slyD encodes a 196 amino acid polypeptide that is a member of the FKBP family of cis–trans peptidyl–prolyl isomerases (PPIases). slyD mutations affect plaque formation by the phage φX174 by blocking the action of the phage lysis protein E. Here we describe the selection of a set of spontaneous slyD mutations conferring resistance to the expression of gene E from a plasmid. These mutations occur disproportionately in residues of SlyD that, based on the structure of the prototype mammalian FKBP12, make ligand contacts with immunosuppressing drug molecules or are conserved in other FKBP proteins. A wide variation in the plating efficiency of φX174 on these E  ^R strains is observed, relative to the parental, indicating that these alleles differ widely in residual SlyD activity. Moreover, it is found that slyD mutations cause significant growth rate defects in Escherichia coli B and C backgrounds. Finally, overexpression of slyD causes filamentation of the host. Thus, among the FKBP genes found in organisms across the evolutionary spectrum, slyD is unique in having three distinct drug-independent phenotypes.     

Identification of the Active Sites in the Methyltransferases of a Transcribing dsRNA Virus

Many double-stranded RNA (dsRNA) viruses are capable of transcribing and capping RNA within a stable icosahedral viral capsid. The turret of turreted dsRNA viruses belonging to the family Reoviridae is formed by five copies of the turret protein, which contains domains with both 7-N-methyltransferase and 2′-O-methyltransferase activities, and serves to catalyze the methylation reactions during RNA capping. Cypovirus of the family Reoviridae provides a good model system for studying the methylation reactions in dsRNA viruses. Here, we present the structure of a transcribing cypovirus to a resolution of ~ 3.8 Å by cryo-electron microscopy. The binding sites for both S-adenosyl-l-methionine and RNA in the two methyltransferases of the turret were identified. Structural analysis of the turret in complex with RNA revealed a pathway through which the RNA molecule reaches the active sites of the two methyltransferases before it is released into the cytoplasm. The pathway shows that RNA capping reactions occur in the active sites of different turret protein monomers, suggesting that RNA capping requires concerted efforts by at least three turret protein monomers. Thus, the turret structure provides novel insights into the precise mechanisms of RNA methylation.     

Pulmonary Toxocariasis Mimicking Invasive Aspergillosis in a Patient with Ulcerative Colitis

A 45-year-old-male who had underlying ulcerative colitis and presented with fever and dry cough. Initially, the patient was considered to have invasive aspergillosis due to a positive galactomannan assay. He was treated with amphotericin B followed by voriconazole. Nevertheless, the patient deteriorated clinically and radiographically. The lung biopsy revealed eosinophilic pneumonia, and ELISA for Toxocara antigen was positive, leading to a diagnosis of pulmonary toxocariasis. After a 10-day treatment course with albendazole and adjunctive steroids, the patient recovered completely without any sequelae. Pulmonary toxocariasis may be considered in patients with subacute or chronic pneumonia unresponsive to antibiotic agents, particularly in cases with eosinophilia.     

Reconsidering connectivity in the sub‐Antarctic

Extreme and remote environments provide useful settings to test ideas about the ecological and evolutionary drivers of biological diversity. In the sub‐Antarctic, isolation by geographic, geological and glaciological processes has long been thought to underpin patterns in the region's terrestrial and marine diversity. Molecular studies using increasingly high‐resolution data are, however, challenging this perspective, demonstrating that many taxa disperse among distant sub‐Antarctic landmasses. Here, we reconsider connectivity in the sub‐Antarctic region, identifying which taxa are relatively isolated, which are well connected, and the scales across which this connectivity occurs in both terrestrial and marine systems. Although many organisms show evidence of occasional long‐distance, trans‐oceanic dispersal, these events are often insufficient to maintain gene flow across the region. Species that do show evidence of connectivity across large distances include both active dispersers and more sedentary species. Overall, connectivity patterns in the sub‐Antarctic at intra‐ and inter‐island scales are highly complex, influenced by life‐history traits and local dynamics such as relative dispersal capacity and propagule pressure, natal philopatry, feeding associations, the extent of human exploitation, past climate cycles, contemporary climate, and physical barriers to movement. An increasing use of molecular data – particularly genomic data sets that can reveal fine‐scale patterns – and more effective international collaboration and communication that facilitates integration of data from across the sub‐Antarctic, are providing fresh insights into the processes driving patterns of diversity in the region. These insights offer a platform for assessing the ways in which changing dispersal mechanisms, such as through increasing human activity and changes to wind and ocean circulation, may alter sub‐Antarctic biodiversity patterns in the future.     

Effect of estradiol and progesterone on long chain fatty acyl-coenzyme A levels in the rat uterus

Fatty acyl-CoAs are potential in vivo inactivators of glucose-6-phosphate dehydrogenase (G6PD). Ovariectomized mature rats (n = 74) were given 5 micrograms of estradiol intravenously, then killed 0, 24, 36, 48 and 72 h later. Control levels of myristoyl-, palmitoyl-, stearoyl-, arachidonoyl-, oleoyl- and linoleoyl-CoA were 0.6, 3.2, 4.7, 3.4, 2.4 and 3.0 micrograms/uterus and were increased 39, 110, 146, 100, 84 and 69% at 36-48 h, respectively. Levels of fatty acyl-CoAs in the rat uterus become elevated 36 h after estradiol treatment. At the same time G6PD changes from a stable enzyme to one that is irreversibly inactivated, possibly due to being rapidly degraded. Progesterone (2 mg subcutaneously every 12 h, n = 30), administered beginning at either 24 or 36 h after estradiol treatment, had no effect on estradiol-induced changes in myristoyl-, palmitoyl-, or stearoyl-CoA. Compared to the groups of rats treated with estradiol alone, animals treated with combinations of estradiol and progesterone exhibited higher levels of arachidonoyl-CoA after 48 h, and oleoyl-CoA and linoleoyl-CoA were greater after 72 h. Progesterone increased the estradiol-induced levels of unsaturated fatty acyl-CoAs suggesting that progesterone may induce uterine fatty acid desaturase activity and/or uptake of dietary fatty acids. Addition of fatty acyl-CoAs, at concentrations seen in vivo at 36-48 h after estradiol, to purified G6PD, causes irreversible G6PD inactivation.     

Rehabilitation and older people.

Rehabilitation is concerned with lessening the impact of disabling conditions. These are particularly common in older people and considerable health gain can be achieved by successful rehabilitation. Hospital doctors and general practitioners should be aware of the core principles of rehabilitation, be able to recognise rehabilitation need in their patients, and have sufficient knowledge of their local rehabilitation services to trigger the referral process.     

萘对川西亚高山森林土壤微生物量、丰度和磷脂脂肪酸的影响

萘作为土壤动物化学抑制剂已在土壤动物生态功能的研究中广泛使用,但其非目标效应使其应用仍存在很大的不确定性。为了解在亚高山森林土壤应用萘抑制土壤动物群落的非目标效应,以川西亚高山森林土壤为研究对象,采用微缩实验研究了土壤微生物生物量、丰度和磷脂脂肪酸对萘胁迫的短期响应。结果表明,萘处理和对照的土壤微生物生物量碳(MBC)、真菌丰度以及细菌、真菌、革兰氏阳性菌(G~+)和革兰氏阴性菌(G~-)PLFAs含量在整个培养期间表现为降低的变化趋势,二者的土壤微生物生物量碳和G~+PLFAs含量以培养52d最低,细菌、真菌和G~-PLFAs含量以培养的45d最低。萘处理和对照的微生物生物量氮(MBN)含量表现出先升高后降低的动态,微生物生物量碳氮比(MBC/MBN)则表现为相反趋势。对照的真菌/细菌PLFAs比值呈现先升高后降低的动态,以培养的17d最高,但萘处理的真菌/细菌PLFAs比值无明显变化规律;萘处理的G~+/G~-PLFAs比值表现为降低的变化趋势,对照的G~+/G~-PLFAs比值表现为先降低后升高的趋势。萘处理仅显著影响了G~+/G~-PLFAs比值,但萘处理和采样时间的交互作用显著影响MBC/MBN、细菌丰度、真菌/细菌丰度比以及细菌、真菌的PLFAs含量、真菌/细菌PLFAs比值、G~+/G~-PLFAs比值。萘作为土壤动物抑制剂对川西亚高山森林土壤微生物群落的非目标效应具有时间变异性。