Genes and genome editing tools for breeding desirable phenotypes in ornamentals.

KEY MESSAGE: We review the main genes underlying commercial traits in cut flower species and critically discuss the possibility to apply genome editing approaches to produce novel variation and phenotypes. Promoting flowering and flower longevity as well as creating novelty in flower structure, colour range and fragrances are major objectives of ornamental plant breeding. The novel genome editing techniques add new possibilities to study gene function and breed new varieties. The implementation of such techniques, however, relies on detailed information about structure and function of genomes and genes. Moreover, improved protocols for efficient delivery of editing reagents are required. Recent results of the application of genome editing techniques to elite ornamental crops are discussed in this review. Enabling technologies and genomic resources are reviewed in relation to the implementation of such approaches. Availability of the main gene sequences, underlying commercial traits and in vitro transformation protocols are provided for the world's best-selling cut flowers, namely rose, lily, chrysanthemum, lisianthus, tulip, gerbera, freesia, alstroemeria, carnation and hydrangea. Results obtained so far are described and their implications for the improvement of flowering, flower architecture, colour, scent and shelf-life are discussed.

Two SERK genes are markers of pluripotency in Cyclamen persicum Mill.

The genetic basis of stem cell specification in somatic embryogenesis and organogenesis is still obscure. SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) genes are involved in embryogenesis and organogenesis in numerous species. In vitro culture of Cyclamen persicum immature ovules provides a system for investigating stem cell formation and maintenance, because lines forming either organs or embryos or callus without organs/embryos are available for the same cultivar and plant growth regulator conditions. The present aim was to exploit this property of cyclamen cultures to understand the role of SERK(s) in stem cell formation and maintenance in somatic embryogenesis and organogenesis in vitro, in comparison with expression in planta. CpSERK1 and CpSERK2 were isolated from embryogenic callus. CpSERK1 and CpSERK2 levels by RT-PCR showed that expression is high in embryogenic, moderate in organogenic, and null in recalcitrant calli. in situ hybridizations showed that the expression of both genes started in clumps of pluripotent stem cells, from which both pre-embryogenic aggregates and organ meristemoids derived, and continued in their trans-amplifying, meristem-like, derivatives. Expression declined in organ meristemoids, in parallel with a partial loss of meristematization. In mature somatic embryos, and in shoot and root primordia, CpSERK1 and CpSERK2 were expressed in meristems, and similar patterns occurred in zygotic embryo and primary meristems in planta. The results point to SERK1 and SERK2 as markers of pluripotency in cyclamen. It is proposed that the high expression of these genes in the trans-amplifying derivatives of the stem cells maintains a pluripotent condition leading to totipotency and, consequently, somatic embryogenesis.

Low-dose cidofovir treatment of BK virus-associated hemorrhagic cystitis in recipients of hematopoietic stem cell transplant.

In recipients of hematopoietic stem cell transplants (HSCTs), BK virus (BKV) has been associated with late-onset hemorrhagic cystitis (HC). In our institution, HSCT recipients with BKV-associated HC are treated with 1 mg/kg of cidofovir weekly. We identified HSCT recipients with BKV-associated HC, treated with weekly cidofovir. Microbiological response was defined as at least a one log reduction in urinary BKV viral load; clinical response was defined as improvement in symptoms and stability or reduction in the grade of cystitis. Nineteen allogeneic HSCT patients received a mean of 4.5 weekly doses of cidofovir. HC occurred at a mean of 68.7 days after transplant. A clinical response was detected in 16/19 (84%) patients, and 9/19 (47%) had a measurable microbiological response (8/10 nonresponders had a BKV viral load above the upper limit of the assay before treatment). Fourteen out of nineteen (74%) patients had no significant increase in serum creatinine. Five patients with renal dysfunction resolved after completion of the therapy and removal of other nephrotoxic agents. We conclude that weekly low-dose cidofovir appears to be a safe treatment option for BKV-associated HC. Although the efficacy of low-dose cidofovir is not proven, a prospective trial is warranted.

Ultraviolet radiation and the risks of cutaneous malignant melanoma and non-melanoma skin cancer: perceptions and behaviours of Danish and American adolescents.

The highest prevalence rates of skin malignancy in the northern hemisphere occur in Scandinavia and the United States (USA). Most Danes and Americans receive 50% of their lifetime ultraviolet (UV) radiation before the age of 21, making it important to address sun exposure risks with adolescents. The project was undertaken to determine differences between Danish and American adolescents in knowledge of sun exposure and skin malignancy, activities accounting for sun exposure, and means used for sun protection. Questionnaires regarding skin cancer and sun exposure were distributed to 674 secondary school age students in Hilleroed, Denmark, and to 483 similarly aged students in Winston-Salem, North Carolina, USA. Differences in responses between and within groups were compared. American adolescents had more knowledge of the characteristics and malignant potential of melanoma than did Danish adolescents. Danish youth and females from both countries were significantly more likely to engage in sunbathing and tanning bed use. Black Danish students reported significantly more sunburn and were more likely to sunbathe or use a tanning bed than were black American students. Danish students were more likely than Americans to use sunscreen, however, Americans were more likely to apply sun protective factor (SPF) 15 or greater. In conclusion, given that sunbathing and tanning bed use are associated with the development of precancerous lesions and skin malignancy, Danish teens are at increased risk. The rates of skin malignancy are relatively high in Scandinavia and efforts to improve understanding of exposure and cancer risks should be undertaken in adolescents.

Validation of cardiologists' decisions to initiate reperfusion therapy for acute myocardial infarction with electrocardiograms viewed on liquid crystal displays of cellular telephones.

BACKGROUND: The transmission of 12-lead electrocardiograms from remote locations to hand-held computers of cardiologists is now possible with the development of wireless technology and computer software. This investigation determined whether the cardiologist's decisions regarding reperfusion therapy for patients with symptoms suggestive of an acute myocardial infarction are the same when given electrocardiograms displayed on a cellular telephone as on a standard paper recording. METHODS: Cardiologists were given 20 electrocardiograms of patients with acute chest pain suggestive of acute myocardial infarction to test the diagnostic reliability of the Nokia 9000i cellular telephone liquid crystal display (LCD) screen. The cardiologists made their decision to initiate or not initiate reperfusion therapy for the patients after viewing their electrocardiograms displayed on both 5-mm and 1-mm formatted grids and twice on traditional printout electrocardiograms. The control level of intraobserver agreement between the responses from the 2 sets of paper display electrocardiograms was compared with the experimental level of intraobserver agreement between the 1-mm LCD electrocardiograms and both sets of paper display electrocardiograms to determine whether the viewing medium affected the cardiologist's decisions. The 1-mm and 5-mm LCD screen electrocardiograms were compared to determine if the grid size affected the cardiologist's decisions. RESULTS: Ninety-three percent of the 2 sets of paper-guided decisions were in agreement. When comparing the 1-mm LCD-guided decisions with both sets of paper-guided decisions, 94% and 89% of the decisions, respectively, were in agreement. The differences between the control and experimental degrees of intraobserver agreement of 1% and 4% were not statistically significant (P1 =.81, P2 =.29). Ninety-one percent of the 1-mm LCD-guided decisions were in agreement with the 5-mm LCD-guided decisions. CONCLUSIONS: Cardiologists' decisions did not vary significantly when viewing either traditional paper electrocardiograms or LCD screen electrocardiograms. Even though there was not a significant difference in the cardiologists' decisions when they viewed electrocardiograms displayed on both the 1-mm and 5-mm grid, it is recommended that the 1-mm grid be used for clinical implementation of the LCD screen.

A community hospital's effort to expedite treatment for patients with chest pain.

OBJECTIVE: The purpose of this study was to determine treatment times at a community hospital that does not receive prehospital electrocardiogram (ECG) transmission and to determine the effect of time to first hospital ECG on overall door-to-drug time. DESIGN: Descriptive. SETTING: 238-bed Regional Medical Center in Burlington, North Carolina. SAMPLE: One hundred four patients with a final diagnosis of acute myocardial infarction were included in this 16-month study. RESULTS: A median door-to-ECG time of 5 minutes was within the American College of Cardiology/American Heart Association recommendation of 10 minutes. Shorter treatment times to obtain the first ECG and initiate thrombolytic therapy were associated with younger patients and those arriving by ambulance. CONCLUSIONS: While efficiency in obtaining a first hospital ECG on patients with suspected acute myocardial infarctions was achieved, this did not result in low door-to-drug times. Further streamlining of protocol and the exploration of prehospital initiatives may result in a significant reduction in door-to-drug times.

Evaluation of the efficacy of hand-held computer screens for cardiologists' interpretations of 12-lead electrocardiograms.

BACKGROUND: Newly designed computer-based applications and the development of wireless technology have allowed the transmission of 12-lead electrocardiogram (ECG) waveforms from remote locations to the hand-held computers of cardiologists. If these computer ECGs can be reliably interpreted, then the time to treatment for cardiac patients may be reduced. METHODS AND RESULTS: Twenty classic examples of cardiac abnormalities were chosen to test the efficacy of the hand-held computer's liquid crystal display (LCD) screen in the interpretation of 12-lead ECGs. Ten cardiologists interpreted these 20 ECGs on the hand-held computers and then twice later on traditional printed paper. The control intraobserver agreement between the sets of paper-displayed ECGs was measured against the agreement between each of the paper sets and the LCD-displayed set of ECGs. Eighty-nine percent (178/200) of the ECGs were interpreted identically by the participants between the 2 paper sets. When comparing the interpretations of the LCD-displayed ECGs with those of each of the paper sets of ECGs, 88.0% (176/200) and 87.5% (175/200) of identical diagnoses were noted. These differences of 1.0% and 1.5% in intraobserver agreement between paper-to-paper and each of the 2 paper-to-LCD comparisons were not significant (P =.75 and P =.88, respectively). CONCLUSIONS: The strong intraobserver agreement shows that cardiologists make the same diagnoses when viewing LCD-displayed ECGs as they do when viewing paper-displayed ECGs. A study to measure the intraobserver agreement of the decision regarding administration of reperfusion therapy after interpretation of ECGs of patients with acute chest pain is now underway.