RESUMO
BACKGROUND: The Amaranthus genus contains at least 20 weedy and invasive species, including Amaranthus palmeri (palmer's amaranth) and Amaranthus tuberculatus (tall waterhemp), two species of regulatory concern in North America, impacting production and yield in crops like corn, soybean and cotton. Amaranthus tuberculatus is regulated in Canada with limited establishment, while current climate models predict a range expansion of A. palmeri impacting crop growing areas in Ontario, Quebec and Manitoba. Since many Amaranthus species are similar in their morphology, especially at the seed stage, this demands the development of additional methods that can efficiently aid in the detection and identification of these species. Protein biotyping using Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) has been traditionally used to identify microorganism species, races and pathotypes. Major protein fractions extracted from an organism, ionized and run through a biotyper using mass spectrometry, result in protein spectra that represent a fingerprint at the species or lower taxonomic rank, providing an efficient molecular diagnostics method. Here we use a modified protein biotyping protocol to extract major protein fractions from seeds of the family Brassicaceae to test our protocol, and then implemented the standardized approach in seeds from Amaranthus species. We then created a database of Amaranthus protein spectra that can be used to test blind samples for a quick identification of species of concern. RESULTS: We generated a protein spectra database with 16 Amaranthus species and several accessions per species, spanning target species of regulatory concern and species which are phylogenetically related or easily confused at the seed stage due to phenotypic plasticity. Testing of two Amaranthus blind sample seed sets against this database showed accuracies of 100% and 87%, respectively. CONCLUSIONS: Our method is highly efficient in identifying Amaranthus species of regulatory concern. The mismatches between our protein biotyping approach and phenotypic identification of seeds are due to absence of the species in the database or close phylogenetic relationship between the species. While A. palmeri cannot be distinguished from A. watsonii, there is evidence these two species have the same native range and are closely related.
RESUMO
Intracardiac echocardiography (ICE) has demonstrated utility in providing high-resolution cardiac ultrasound images for guidance of numerous catheter-based interventions, including radiofrequency ablations (RFA). However, the training of interventionalists and refinement of procedures involving intracardiac catheters is costly and time consuming due to necessary clinical and animal studies. As a result, research and development of ICE for other purposes is gradual and deliberate. Intracardiac acoustic radiation force impulse (ARFI) imaging has been demonstrated to be a suitable modality to monitor the progress of RFA procedures; however, a clinical protocol has been slow to develop due to the expense and demands of clinical experiments. We report on the development and use of an ex vivo heart model to evaluate ICE and intracardiac ARFI imaging. The ability of this model to provide clinically-relevant intracardiac imaging angles was investigated by inserting an intracardiac probe into the heart and imaging it from various positions and orientations. ARFI images of all four chambers also were formed. RFAs were also performed to create stiffer lesions within the right and left ventricles. Upon completion of the ablation, ARFI imaging was used to visualize the lesion and compared with images taken from pathology.The results show the ovine heart model to be a suitable apparatus for recreating several clinically-relevant intracardiac viewing angles of the heart. Also, the results indicate the potential of the heart model to be a valuable tool in the future development and refinement of a clinical protocol for intracardiac ARFI imaging based guidance and assessment of cardiac radiofrequency ablations.
