Investigation of the reaction of magnesium with p-nitro-o-carboxylazochromotropic acid and the determination of magnesium in plants and in water

The formation of a novel complex between magnesium and p-nitro-o-carboxylazochromotropic acid (NCBAC) has been observed which is sensitive and selective at pH 10.4 in the presence of ethylenediamine and fluoride. The constants of formation of this complex have been determined by a spectral correction technique, and the complexation reaction has been applied to the spectrophotometric determination of trace magnesium in plants and in water with satisfactory results.     

Towards therapeutic applications of engineered zinc finger proteins

It has long been the goal of molecular biologists to design DNA-binding proteins for the specific control of gene expression. The zinc finger design is ideally suited for such purposes, discriminating between closely related sequences both in vitro and in vivo. Whereas other DNA-binding proteins generally make use of the 2-fold symmetry of the double helix, zinc fingers do not and so can be linked linearly in tandem to recognize DNA sequences of different lengths, with high fidelity. This modular design offers a large number of combinatorial possibilities for the specific recognition of DNA. By fusing zinc finger peptides to repression or activation domains, genes can be selectively targeted and switched off and on. Several recent applications of such engineered zinc finger proteins (ZFPs) are described, including the activation of vascular endothelial growth factor (VEGF) in a human cell line and an animal model. Clinical trials have recently begun on using VEGF-activating ZFPs to treat human peripheral arterial disease, by stimulating vascular growth. Also in progress are pre-clinical studies using ZFPs to target the defective genes in two monogenic disorders, SCID and SCA. The aim is to replace them in each case by a correct copy from an extrachromosomal DNA donor by means of homologous recombination. Promising results are reported.     

Estimating local interaction from spatiotemporal forest data, and Monte Carlo bias correction

We point out a general problem in fitting continuous time spatially explicit models to a temporal sequence of spatial data observed at discrete times. To illustrate the problem, we examined the continuous time Markov model for forest gap dynamics. A forest is assumed to be apportioned into discrete cells (or sites) arranged in a regular square lattice. Each site is characterized as either a gap or a non-gap site according to the vegetation height of trees. The model incorporates the influence of neighboring sites on transition rate: transition rate from a non-gap to a gap site increases linearly with the number of neighbors that are currently in the gap state, and vice versa. We fitted the model to the spatiotemporal data of canopy height observed at the permanent plot in Barro Colorado Island (BCI). When we used the approximate maximum likelihood method to estimate the parameters of the model, the estimated transition rates included a large bias-in particular, the strength of interaction between nearby sites was underestimated. This bias originated from the assumption that each transition between two observation times is independent. The interaction between sites at local scale creates a long chain of transitions within a single census interval, which violates the independence of each transition. We show that a computer-intensive method, called Monte Carlo bias correction (MCBC), is very effective in removing the bias included in the estimate. The global and local gap densities measuring spatial aggregation of gap sites were computed from simulated and real gap dynamics to assess the model. When the approximate likelihood estimates were applied to the model, the predicted local gap density was clearly lower than the observed one. The use of MCBC estimates, suggesting a strong interaction between sites, improved this discrepancy.     

FEEDING METHOD AFFECTS OTOLITH DIGESTION IN CAPTIVE GRAY SEALS: IMPLICATIONS FOR DIET COMPOSITION ESTIMATION

Analysis of prey structures from scats has become widely used for assessing the diet of free-ranging seals. Captive feeding experiments have played an important role in the development of scat analysis but methodology has varied. We investigated whether using an experimental otolith-carrier species or in situ otolith experimental prey to feed haddock, plaice, and sand eel otoliths to two captive gray seals affected the amount of otolith digestion, and therefore our ability to estimate fish size and diet composition. Otolith recovery rates varied among the three prey species, but were not affected by feeding method. Otoliths were more digested in carrier experiments; resulting digestion coefficients were greater than those from in situ experiments. As a result, while the lengths of whole fish fed experimentally could be accurately predicted by applying in situ digestion coefficients, the lengths of these fish could not be accurately predicted using carrier digestion coefficients. Consequently, the estimated proportion by weight of each species in the diet can change markedly when carrier, as opposed to in situ , digestion coefficients are used. The implications for diet composition estimation are significant.     

Taux de fixation de la pointe du ventricule gauche et correction d’atténuation TEMP/TDM dans la scintigraphie myocardique de perfusion chez les patients non ischémiques

Introduction

Attenuation correction (AC) has been shown to improve the accuracy of myocardial perfusion single photon emission computed scintigraphy (SPECT). It often reveals a decrease in apical uptake in the AC of the left ventricular (LV) in non-ischemic patients. We aimed to identify the parameters that could affect the apical radiotracer uptake in non-ischemic patients.

PARP-mediated repair,homologous recombination,and back-up non-homologous end joining-like repair of single-strand nicks

Double-strand breaks (DSBs) in chromosomal DNA can induce both homologous recombination (HR) and non-homologous end-joining (NHEJ). Recently we showed that single-strand nicks induce HR with a significant reduction in toxicity and mutagenic effects associated with NHEJ. To further investigate the differences and similarities of DSB- and nick-induced repair, we used an integrated reporter system in human cells to measure HR and NHEJ produced by the homing endonuclease I-AniI and a designed ‘nickase’ variant that nicks the same target site, focusing on the PARP and HR repair pathways. PARP inhibitors, which block single-strand break repair, increased the rate of nick-induced HR up to 1.7-fold but did not affect DSB-induced HR or mutNHEJ. Additionally, expression of the PALB2 WD40 domain in trans acted as a dominant-negative inhibitor of both DSB- and nick-induced HR, sensitized cells to PARP inhibition, and revealed an alternative mutagenic repair pathway for nicks. Thus, while both DSB- and nick-induced HR use a common pathway, their substrates are differentially processed by cellular factors. These results also suggest that the synthetic lethality of PARP and BRCA may be due to repair of nicks through an error prone, NHEJ-like mechanism that is active when both PARP and HR pathways are blocked.     

Considerations for normalization of DNA methylation data by Illumina 450K BeadChip assay in population studies

Analysis of epigenetic mechanisms, particularly DNA methylation, is of increasing interest for epidemiologic studies examining disease etiology and impacts of environmental exposures. The Infinium HumanMethylation450 BeadChip^® (450K), which interrogates over 480?000 CpG sites and is relatively cost effective, has become a popular tool to characterize the DNA methylome. For large-scale studies, minimizing technical variability and potential bias is paramount. The goal of this paper was to evaluate the performance of several existing and novel color channel normalizations designed to reduce technical variability and batch effects in 450K analysis from a large population study. Comparative assessment of 10 normalization procedures included the GenomeStudio^® Illumina procedure, the lumi smooth quantile approach, and the newly proposed All Sample Mean Normalization (ASMN). We also examined the performance of normalizations in combination with correction for the two types of Infinium chemistry utilized on the 450K array. We observed that the performance of the GenomeStudio^® normalization procedure was highly variable and dependent on the quality of the first sample analyzed in an experiment, which is used as a reference in this procedure. While the lumi normalization was able to decrease batch variability, it increased variation among technical replicates, potentially reducing biologically meaningful findings. The proposed ASMN procedure performed consistently well, both at reducing batch effects and improving replicate comparability. In summary, the ASMN procedure can improve existing color channel normalization, especially for large epidemiologic studies, and can be successfully implemented to enhance a 450K DNA methylation data pipeline.     

A Pragmatic Approach for Determining Otter Distribution from Disparate Occurrence Records

Opportunistic records of animal occurrence may be problematic for inferring species distribution and habitat requirements because of unknown and uncontrolled sources of sampling variance. In this study, we used occurrence records for river otters (Lontra canadensis) derived from sign surveys, road kills, trapper bycatch, and opportunistic sightings (n = 185 records collected 2001–2012) to assess the potential distribution and habitat relationships of otters across central and western New York, USA. To mitigate for obvious observation biases, we standardized observation intensity across regions a priori and restricted inference to readily accessible areas (i.e., ≤700 m from the nearest road). Model selection, and the direction of covariate effects, proved robust to these sampling biases although effect sizes varied −7.1% to +48.0% after bias correction, with the coefficient for the proportion of available shoreline being the most unstable. Ultimately, the top bias-corrected model proved a reliable index for otter probability of occurrence given a strong, positive, and linear relationship with a withheld set of standardized survey records for otters collected in winter 2016–2017 (n = 57; R² = 0.90). This model indicated that approximately 20% of the study area represented high probability of otter occurrence. We demonstrated that reliable inference on wildlife habitat requirements can be obtained from disparate records of animal occurrence provided that data biases are known and effectively mitigated. © 2020 The Wildlife Society.