Deuterated nucleotides as chemical probes of RNA structure: a detailed protocol for the enzymatic synthesis of a complete set of nucleotides specifically deuterated at ribose carbons

We describe here a detailed protocol for the synthesis of ribonucleotides specifically deuterated at each ribose carbon atom. We synthesized 20 specifically deuterated ribonucleotides: ATP, CTP, GTP, and UTP, each of which contained one of five deuterated riboses (either 1′-D, 2″-D, 3′-D, 4′-D, or 5′,5″-D2). Our synthetic approach is inspired by the pioneering work of Tolbert and Williamson, who developed a method for the convenient one-pot enzymatic synthesis of nucleotides (Tolbert, T. J. and Williamson, J. R. (1996) J. Am. Chem. Soc. 118, 7929–7940). Our protocol consists of a comprehensive list of required chemical and enzymatic reagents and equipment, detailed procedures for enzymatic assays and nucleotide synthesis, and chromatographic procedures for purification of deuterated nucleotides. As an example of the utility of specifically deuterated nucleotides, we used them to synthesize specifically deuterated sarcin/ricin loop (SRL) RNA and measured the deuterium kinetic isotope effect on hydroxyl radical cleavage of the SRL.https://www.scienceopen.com/document/read?vid=eb44f1b0-c408-4336-a2c0-aed203250898Published versio

Results of the tagging of salt water fishes in Florida

The purpose of this paper is to report the results to date (October 1, 1961) of the 1960-61 fish-tagging program conducted in Zone I (Northern Hernando County Line to Everglades City)along the southwest coast of Florida and Zone II (Everglades City to the northern Indian River County Line.) (66pp.

The window of desiccation tolerance shown by early-stage germinating seedlings remains open in the resurrection plant, Xerophyta viscosa

Resurrection plants are renowned for their vegetative desiccation tolerance (DT). While DT in vegetative tissues is rare in angiosperms, it is ubiquitous in mature orthodox seeds. During germination, seedlings gradually lose DT until they pass a point of no return, after which they can no longer survive dehydration. Here we investigate whether seedlings of the resurrection plant Xerophyta viscosa ever lose the capacity to establish DT. Seedlings from different stages of germination were dehydrated for 48 hours and assessed for their ability to recover upon rehydration. While a transient decline in the ability of X. viscosa seedlings to survive dehydration was observed, at no point during germination was the ability to re-establish DT completely lost in all seedlings. Pre-treatment of seedlings with PEG or sucrose reduced this transient decline, and improved the survival rate at all stages of germination. Additionally, we observed that the trait of poikilochlorophylly (or loss of chlorophyll) observed in adult X. viscosa leaves can be induced throughout seedling development. These results suggest that the window of DT seen in germinating orthodox seeds remains open in X. viscosa seedlings and that vegetative DT in Xerophyta species may have evolved from the ability to retain this program through to adulthood

Changing Spatial Distribution of the Church of the Nazarene

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Stress in the Patella Following Autologous Chondrocyte Implantation - A Finite Element Study

Bovine patella cartilage shows signs of damage and cell death when subjected to a compressive cyclic load of 6 MPa, which results in a shear stress of 5.6 MPa. The aim of this research was to investigate the effect of activities of daily living (descending stairs, bicycling and deep flexion) on the contact stresses in the patellofemoral compartment following an articular chondrocyte implantation (ACI). A finite element (FE) model of the patellar femoral joint was created and dynamic non-linear analyses were carried out for this purpose. A shear stress of 5.6 MPa was used as the threshold that cartilage can tolerate without resulting in damage. The FE model was verified numerically. Our results show that, for a 70 kg individual at 50% recovery, (i) contact stress in the patella is 11% higher than that in the femur; (ii) shear stress in the host cartilage reaches 4.75 MPa at 50° of flexion; (iii) shear stress in the patella host cartilage is twice that in a healthy cartilage during deep flexion approaching 70°; (iv) maximum shear stress value was 2.75 MPa during cycling at 60% load; (v) stress shielding still occurs through the host cartilage even when the implanted cartilage reaches 97.5% the Young’s modulus of a healthy cartilage. Based on these results, (i) using an exercise bicycle is recommended for rehabilitation; (ii) deep knee flexion should be avoided; (iii) obese people with a BMI of over 42 kg/m2 should not undertake vigorous weight-bearing exercises involving deep knee flexion

A Memorial for William V. Sliter

William V. (Bill) Sliter, an internationally known micropaleontologist and research geologist for the U.S. Geological Survey, passed away suddenly, October, 1997, while talking to a colleague in his office. In his honor, B. Huber, T. Bralower, and M. Leckie organized a keynote symposium ‘‘Paleoecological and Geochemical Signatures of Cretaceous Anoxic Events’’ at the 1998 annual meeting of the Geological Society of America in Toronto, Canada. This theme issue of the Journal of Foraminiferal Research contains the published papers from the symposium and is dedicated to his memory

A Memorial for William V. Sliter

William V. (Bill) Sliter, an internationally known micropaleontologist and research geologist for the U.S. Geological Survey, passed away suddenly, October, 1997, while talking to a colleague in his office. In his honor, B. Huber, T. Bralower, and M. Leckie organized a keynote symposium ‘‘Paleoecological and Geochemical Signatures of Cretaceous Anoxic Events’’ at the 1998 annual meeting of the Geological Society of America in Toronto, Canada. This theme issue of the Journal of Foraminiferal Research contains the published papers from the symposium and is dedicated to his memory

Extending Aromatase-Inhibitor Adjuvant Therapy to 10 Years

Treatment with an aromatase inhibitor for 5 years as up-front monotherapy or after tamoxifen therapy is the treatment of choice for hormone-receptor–positive early breast cancer in postmenopausal women. Extending treatment with an aromatase inhibitor to 10 years may further reduce the risk of breast-cancer recurrence

Increased resistance to biotrophic pathogens in the Arabidopsis constitutive induced resistance 1 mutant is EDS1 and PAD4-dependent and modulated by environmental temperature

The Arabidopsis constitutive induced resistance 1 ( cir1 ) mutant displays salicylic acid (SA)-dependent constitutive expression of defence genes and enhanced resistance to biotrophic pathogens. To further characterise the role of CIR1 in plant immunity we conducted epistasis analyses with two key components of the SA-signalling branch of the defence network, ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1) and PHYTOALEXIN DEFICIENT4 (PAD4). We demonstrate that the constitutive defence phenotypes of cir1 require both EDS1 and PAD4, indicating that CIR1 lies upstream of the EDS1-PAD4 regulatory node in the immune signalling network. In light of this finding we examined EDS1 expression in cir1 and observed increased protein, but not mRNA levels in this mutant, suggesting that CIR1 might act as a negative regulator of EDS1 via a post-transcriptional mechanism. Finally, as environmental temperature is known to influence the outcome of plant-pathogen interactions, we analysed cir1 plants grown at 18, 22 or 25°C. We found that susceptibility to Pseudomonas syringae pv. tomato ( Pst ) DC3000 is modulated by temperature in cir1 . Greatest resistance to this pathogen (relative to PR-1:LUC control plants) was observed at 18°C, while at 25°C no difference in susceptibility between cir1 and control plants was apparent. The increase in resistance to Pst DC3000 at 18°C correlated with a stunted growth phenotype, suggesting that activation of defence responses may be enhanced at lower temperatures in the cir1 mutant