Safety assessment of food and feed from biotechnology-derived crops employing RNA-mediated gene regulation to achieve desired traits: A scientific review

Gene expression can be modulated in plants to produce desired traits through agricultural biotechnology. Currently, biotechnology-derived crops are compared to their conventional counterparts, with safety assessments conducted on the genetic modification and the intended and unintended differences. This review proposes that this comparative safety assessment paradigm is appropriate for plants modified to express mediators of RNA-mediated gene regulation, including RNA interference (RNAi), a gene suppression mechanism that naturally occurs in plants and animals. The molecular mediators of RNAi, including long double-stranded RNAs (dsRNA), small interfering RNAs (siRNA), and microRNAs (miRNA), occur naturally in foods; therefore, there is an extensive history of safe consumption. Systemic exposure following consumption of plants containing dsRNAs that mediate RNAi is limited in higher organisms by extensive degradation of ingested nucleic acids and by biological barriers to uptake and efficacy of exogenous nucleic acids. A number of mammalian RNAi studies support the concept that a large margin of safety will exist for any small fraction of RNAs that might be absorbed following consumption of foods from biotechnology-derived plants that employ RNA-mediated gene regulation. Food and feed derived from these crops utilizing RNA-based mechanisms is therefore expected to be as safe as food and feed derived through conventional plant breeding.     

Perceptions of Home Dialysis Training and Experience Among US Nephrology Fellows

1. Download : Download high-res image (397KB)
2. Download : Download full-size image

     

Circadian Rhythmicity as a Predictor of Quality of Life in Allogeneic Hematopoietic Cell Transplant Patients

Context

Quality of life (QoL) is increasingly recognized as an important outcome of cancer treatment. Previous studies have examined clinical predictors of QoL, but with the increasing prevalence of wearable sensors that monitor sleep and activity patterns, further investigation into whether these behaviors are predictive of post-treatment QoL is now feasible. Among patients receiving aggressive cancer treatment such as hematopoietic cell transplantation (HCT), analysis of circadian rhythms (24-hour patterns of sleep and activity) via wearable sensors is limited.

Stem water cryogenic extraction biases estimation in deuterium isotope composition of plant source water

The hydrogen isotope ratio of water cryogenically extracted from plant stem samples (δ²H_{stem_CVD}) is routinely used to aid isotope applications that span hydrological, ecological, and paleoclimatological research. However, an increasing number of studies have shown that a key assumption of these applications—that δ²H_{stem_CVD} is equal to the δ²H of plant source water (δ²H_source)—is not necessarily met in plants from various habitats. To examine this assumption, we purposedly designed an experimental system to allow independent measurements of δ²H_{stem_CVD}, δ²H_source, and δ²H of water transported in xylem conduits (δ²H_xylem) under controlled conditions. Our measurements performed on nine woody plant species from diverse habitats revealed a consistent and significant depletion in δ²H_{stem_CVD} compared with both δ²H_source and δ²H_xylem. Meanwhile, no significant discrepancy was observed between δ²H_source and δ²H_xylem in any of the plants investigated. These results cast significant doubt on the long-standing view that deuterium fractionation occurs during root water uptake and, alternatively, suggest that measurement bias inherent in the cryogenic extraction method is the root cause of δ²H_{stem_CVD} depletion. We used a rehydration experiment to show that the stem water cryogenic extraction error could originate from a dynamic exchange between organically bound deuterium and liquid water during water extraction. In light of our finding, we suggest caution when partitioning plant water sources and reconstructing past climates using hydrogen isotopes, and carefully propose that the paradigm-shifting phenomenon of ecohydrological separation (“two water worlds”) is underpinned by an extraction artifact.

The analysis of the stable isotope ratios of plant source water (δ_source) is a powerful tool enabling the elucidation of a range of plant physiological, ecological, and hydrological processes from scales ranging from individual plants to the planet. δ_source provides a foundation on which to form isotope signals of transpired water vapor and plant-derived biomarkers (i.e., cellulose and lipids) and thus is of high relevance to studies of terrestrial water fluxes (1, 2) and paleoclimate reconstructions (3, 4). δ_source also contains information on the spatial and temporal origins of water used by plants and so is commonly used for investigating plant water uptake patterns under natural conditions (5, 6). Moreover, dual-isotope (δ²H and δ¹⁸O) analysis of δ_source was critical in formulating the paradigm-shifting “two water worlds” (TWW) hypothesis, whereby ecohydrological separation exists between plant-accessible soil water pools and those recharging streams and groundwater (7, 8).Elucidation of the foregoing processes rest on the assumption that water extracted from plant stems is isotopically identical to water taken up by plant roots. Plant stem water is typically extracted with the cryogenic-vacuum distillation technique; δ generated with this method is hereinafter referred as δ_{stem_CVD} (9). For δ_{stem_CVD} to be an accurate indicator of δ_source (i.e., δ_{stem_CVD} = δ_source), two prerequisites must be met: 1) isotope change does not occur during root uptake and/or xylem transport of the source water (prerequisite I) and 2) stem water cryogenic extraction is a robust approach toward isotope recovery of xylem water (prerequisite II). The “δ_{stem_CVD} = δ_source” assumption is generally valid for oxygen isotopes of water, but numerous studies have used hydrogen isotopes to assess source water, and here this assumption has faced scrutiny, as multiple studies have reported significant depletion in δ²H_{stem_CVD} compared with δ²H_source in plants from various habitats (10–18).A frequently invoked explanation for the observed δ²H_{stem_CVD} depletion is a violation of prerequisite I, as it is believed that symplastic uptake of source water into the root xylem can give rise to hydrogen isotope fractionation (10, 11, 13, 19). The available evidence (10, 11) in support of such an explanation is largely peripheral, because direct, unambiguous confirmation of water uptake/transport-related fractionation would require a comparison of deuterium in source water and water transported within xylem conduits (δ²H_xylem). However, this type of comparison is difficult owing to the technical challenges in obtaining targeted measurements of δ²H_xylem in most plants. Intriguingly, in a field-grown riparian tree species (Populus euphratica) in which δ²H_xylem measurement was made possible with the aid of a syringe-aided xylem sap bleeding technique, no significant difference was observed between δ²H_xylem and δ²H_source (12). This led to the suggestion that, at least for the investigated species, δ²H_{stem_CVD} depletion arises not from a violation of prerequisite I, but rather from a violation of prerequisite II. The violation of prerequisite II has been deemed possible (12, 17) based on the argument that hydrogen isotope heterogeneity could be present within the bulk stem water (i.e., the outside xylem water may carry a metabolism-induced, more-depleted δ²H signature compared with the xylem water), potentially causing the stem water extraction technique to artifactually underestimate δ²H_xylem.Given the controvertible state of knowledge regarding the mechanism driving δ²H_{stem_CVD} depletion, it is imperative for us to build a better and more comprehensive understanding of the isotopic relationships among cryogenic extracted bulk stem water, source water, and xylem water in different plants, so as to put the application of the stem water cryogenic extraction technique in diverse fields on firmer ground. In this context, it should be pointed out that the xylem water direct sampling technique (12) is applicable only to a few riparian tree species. Recently, a new method relying on laser-enabled isotope measurement of water vapor in equilibrium with xylem water has demonstrated potential for in situ continuous monitoring of xylem isotope signatures in trees (20, 21); however, the method needs further development before it becomes broadly applicable to different plant types. Thus, a more generally applicable method is needed for determining xylem water signature across diverse plant types.Toward this goal, and capitalizing on the well-recognized mass balance-dictated principle that the isotopic composition of steady-state (SS) plant transpiration is identical to that of the xylem water supplying the plant canopy, we custom-designed a measurement system to enable independent quantification of xylem water isotope composition through isotope measurement of SS plant transpiration. This measurement system conferred the ability to compare values of δ_{stem_CVD}, δ_source, and δ_xylem across a number of plant species of varying native habitats. The data allowed us to confirm the common presence of δ²H_{stem_CVD} depletion across all plant types measured, and also to demonstrate that this phenomenon is caused by cryogenic extraction-associated artifact and not by water uptake/transport-related fractionation. We also performed a rehydration experiment to illustrate that the extraction artifact is unrelated to within-stem isotope heterogeneity as has been recently suggested, but rather is more likely linked to a deuterium-exchange process that occurs dynamically during cryogenic extraction. Using the TWW hypothesis as an example, we further discuss the ramifications for ecological/hydrological queries that rely on accurate isotopic information on plant source/xylem water.     

The Toxicology Investigators Consortium Case Registry—the 2014 Experience

The Toxicology Investigators Consortium (ToxIC) Case Registry was established in 2010 by the American College of Medical Toxicology. The Registry includes all medical toxicology consultations performed at participating sites. The Registry was queried for all cases entered between January 1 and December 31, 2014. Specific data reviewed for analysis included demographics (age, gender, ethnicity), source of consultation, reasons for consultation, agents involved in toxicological exposures, signs, symptoms, clinical findings, fatalities, and treatment. In 2014, 9172 cases were entered in the Registry across 47 active member sites. Females accounted for 51.1 % of cases. The majority (65.1 %) of cases were adults between the ages of 19 and 65. Caucasians made up the largest identified ethnic group (48.9 %). Most Registry cases originated from the inpatient setting (93.5 %), with a large majority of these consultations coming from the emergency department or inpatient admission services. Intentional and unintentional pharmaceutical exposures continued to be the most frequent reasons for consultation, accounting for 61.7 % of cases. Among cases of intentional pharmaceutical exposure, 62.4 % were associated with a self-harm attempt. Non-pharmaceutical exposures accounted for 14.1 % of Registry cases. Similar to the past years, non-opioid analgesics, sedative-hypnotics, and opioids were the most commonly encountered agents. Clinical signs or symptoms were noted in 81.9 % of cases. There were 89 recorded fatalities (0.97 %). Medical treatment (e.g., antidotes, antivenom, chelators, supportive care) was rendered in 62.3 % of cases. Patient demographics and exposure characteristics in 2014 Registry cases remain similar to prior years. The majority of consultations arose in the acute care setting (emergency department or inpatient) and involved exposures to pharmaceutical products. Among exposures, non-opioid analgesics, sedative/hypnotics, and opioids were the most frequently encountered. A majority of cases required some form of treatment, but fatalities were rare.

Electronic supplementary material

The online version of this article (doi:10.1007/s13181-015-0507-7) contains supplementary material, which is available to authorized users.     

Development of a parent-proxy quality-of-life chronic cough-specific questionnaire: clinical impact vs psychometric evaluations

BACKGROUND: Chronic cough affects at least 7% of children, and the impact of this on families is significant. Although adult cough-specific quality-of-life (QOL) instruments have been shown to be a useful cough outcome measure, no suitable cough-specific QOL for parents of children with chronic cough exists. This article compares two methods of item reduction (clinical impact and psychometric) and reports on the statistical properties of both QOL instruments. METHOD: One hundred seventy children (97 boys and 73 girls; median age, 4 years; interquartile range, 3 to 7.25 years) and one of their parents participated. A preliminary 50-item parent cough-specific QOL (PC-QOL) questionnaire was developed from conversations with parents of children with chronic cough (ie, cough for > 3 weeks). Parents also completed generic QOL questionnaires (eg, Pediatric Quality of Life Inventory, version 4.0 [PedsQL4.0] and the 12-item Short Form Health Survey, version 2 [SF-12v2]). RESULTS: The clinical impact and psychometric method of item reduction resulted in 27-item and 26-item PC-QOL questionnaires, respectively, with approximately 50% of items overlapping. Internal consistency among the final items from both methods was excellent. Some evidence for concurrent and criterion validity of both methods was established as significant correlations were found between subscales of the PC-QOL questionnaire and the scales of the SF-12v2 and PedsQL4.0 scores. The PC-QOL questionnaire derived from both methods was sensitive to change following an intervention. CONCLUSION: Chronic cough significantly impacts on the QOL of both parents and children. Although the PC-QOL questionnaires derived from a clinical impact method and from a psychometric method contained different items, both versions were shown to be internally consistent and valid. Further testing is required to compare both final versions to objective and subjective cough measures.     

Regenerative peripheral nerve interfaces for real-time,proportional control of a Neuroprosthetic hand

Introduction

Regenerative peripheral nerve interfaces (RPNIs) are biological constructs which amplify neural signals and have shown long-term stability in rat models. Real-time control of a neuroprosthesis in rat models has not yet been demonstrated. The purpose of this study was to: a) design and validate a system for translating electromyography (EMG) signals from an RPNI in a rat model into real-time control of a neuroprosthetic hand, and; b) use the system to demonstrate RPNI proportional neuroprosthesis control.

Methods

Animals were randomly assigned to three experimental groups: (1) Control; (2) Denervated, and; (3) RPNI. In the RPNI group, the extensor digitorum longus (EDL) muscle was dissected free, denervated, transferred to the lateral thigh and neurotized with the residual end of the transected common peroneal nerve. Rats received tactile stimuli to the hind-limb via monofilaments, and electrodes were used to record EMG. Signals were filtered, rectified and integrated using a moving sample window. Processed EMG signals (iEMG) from RPNIs were validated against Control and Denervated group outputs.

Results

Voluntary reflexive rat movements produced signaling that activated the prosthesis in both the Control and RPNI groups, but produced no activation in the Denervated group. Signal-to-Noise ratio between hind-limb movement and resting iEMG was 3.55 for Controls and 3.81 for RPNIs. Both Control and RPNI groups exhibited a logarithmic iEMG increase with increased monofilament pressure, allowing graded prosthetic hand speed control (R²?=?0.758 and R²?=?0.802, respectively).

Conclusion

EMG signals were successfully acquired from RPNIs and translated into real-time neuroprosthetic control. Signal contamination from muscles adjacent to the RPNI was minimal. RPNI constructs provided reliable proportional prosthetic hand control.