Effects of the Use of a Mallet during Bone Marrow Biopsy Collection on Specimen Quality: A Quality Improvement Project.

CONTEXT: Avoiding procedure-related morphologic distortion such as fragmentation and crush artifact is critical in bone marrow diagnosis. Use of a hammer or mallet, although infrequent, is a known technique of advancing the biopsy needle during specimen collection. OBJECTIVES: We performed a double-blinded, retrospective review of bone marrow biopsies collected by the Interventional Radiology department at our institution in order to assess specimen quality by using this technique. DESIGN: We reviewed 93 bone marrow biopsy specimens collected at our hospitals, between January 2015 and June 2015. Routine bone marrow core biopsy slides were reviewed. The presence of crush artifact, specimen fragmentation, and aspiration artifact, as well as the presence of osteopenia and an overall grade of specimen adequacy, was recorded for each specimen. RESULTS: A sterile mallet was used during the bone marrow biopsy procedure in 29 cases. Use of a mallet was significantly associated with the presence of suboptimal or inadequate specimen quality of bone marrow core biopsy (p<0.005) and was independently associated with severe specimen fragmentation (2+) (p<0.0001). There was no statistically significant association between length of the core and use of a mallet. CONCLUSIONS: Use of a mallet during bone marrow core biopsy collection is significantly associated with morphologic distortion in the form of severe specimen fragmentation and negatively affects specimen adequacy. There is no difference in length of core biopsy as previously thought by using a mallet to advance the needle during the procedure. We recommend that the use of this technique should be avoided during specimen collection.

Ultra-fast alterations in mRNA levels uncover multiple players in light stress acclimation in plants.

The acclimation of plants to changes in light intensity requires rapid responses at several different levels. These include biochemical and biophysical responses as well as alterations in the steady-state level of different transcripts and proteins. Recent studies utilizing promoter::reporter constructs suggested that transcriptional responses to changes in light intensity could occur within seconds, rates for which changes in mRNA expression are not routinely measured or functionally studied. To identify and characterize rapid changes in the steady-state level of different transcripts in response to light stress we performed RNA sequencing analysis of Arabidopsis thaliana plants subjected to light stress. Here we report that mRNA accumulation of 731 transcripts occurs as early as 20-60 sec following light stress application, and that at least five of these early response transcripts play an important biological role in the acclimation of plants to light stress. More than 20% of transcripts accumulating in plants within 20-60 sec of initiation of light stress are H2 O2 - and ABA-response transcripts, and the accumulation of several of these transcripts is inhibited by transcriptional inhibitors. In accordance with the association of rapid response transcripts with H2 O2 and ABA signaling, a mutant impaired in ABA sensing (abi-1) was found to be more tolerant to light stress, and the response of several of the rapid response transcripts was altered in mutants impaired in reactive oxygen metabolism. Our findings reveal that transcriptome reprogramming in plants could occur within seconds of initiation of abiotic stress and that this response could invoke known as well as unknown proteins and pathways.

ROS as key players in plant stress signalling.

Reactive oxygen species (ROS) play an integral role as signalling molecules in the regulation of numerous biological processes such as growth, development, and responses to biotic and/or abiotic stimuli in plants. To some extent, various functions of ROS signalling are attributed to differences in the regulatory mechanisms of respiratory burst oxidase homologues (RBOHs) that are involved in a multitude of different signal transduction pathways activated in assorted tissue and cell types under fluctuating environmental conditions. Recent findings revealed that stress responses in plants are mediated by a temporal-spatial coordination between ROS and other signals that rely on production of stress-specific chemicals, compounds, and hormones. In this review we will provide an update of recent findings related to the integration of ROS signals with an array of signalling pathways aimed at regulating different responses in plants. In particular, we will address signals that confer systemic acquired resistance (SAR) or systemic acquired acclimation (SAA) in plants.

Transposon tagging in diploid strawberry.

Fragaria vesca was transformed with a transposon tagging construct harbouring amino terminally deleted maize transposase and EGFP (Ac element), NPTII, CaMV 35S promoter (P35S) driving transposase and mannopine synthase promoter (Pmas) driving EGFP (Ds element). Of 180 primary transgenics, 48 were potential launch pads, 72 were multiple insertions or chimaeras, and 60 exhibited somatic transposition. T1 progeny of 32 putative launch pads were screened by multiplex PCR for transposition. Evidence of germ-line transposition occurred in 13 putative launch pads; however, the transposition frequency was too low in three for efficient recovery of transposants. The transposition frequency in the remaining launch pads ranged from 16% to 40%. After self-pollination of the T0 launch pads, putative transposants in the T1 generation were identified by multiplex PCR. Sequencing of hiTAIL-PCR products derived from nested primers within the Ds end sequences (either P35S at the left border or the inverted repeat at the right border) of T1 plants revealed transposition of the Ds element to distant sites in the strawberry genome. From more than 2400 T1 plants screened, 103 unique transposants have been identified, among which 17 were somatic transpositions observed in the T0 generation. Ds insertion sites were dispersed among various gene elements [exons (15%), introns (23%), promoters (30%), 3' UTRs (17%) as well as intergenically (15%)]. Three-primer (one on either side of the Ds insertion and one within the Ds T-DNA) PCR could be used to identify homozygous T2 transposon-tagged plants. The mutant collection has been catalogued in an on-line database.

Transcatheter arterial embolization for the palliation of painful bone lesions.

Pain related to bone tumors, whether benign or malignant, can be significantly debilitating to patients. Unfortunately, there is no single optimal treatment solution for tumor-related bone pain. Many treatment options exist for the palliation of bone pain, each with their own advantages and disadvantages. This article discusses the use of bland arterial embolization for the purpose of pain control in patients with both primary and metastatic bone tumors. Considerations for patient selection and preparation, procedural steps, overcoming technical challenges, potential complications, and follow-up care will be reviewed using case examples.

Development of efficient plant regeneration and transformation system for impatiens using Agrobacterium tumefaciens and multiple bud cultures as explants.

BACKGROUND: Impatiens (Impatiens walleriana) is a top selling floriculture crop. The potential for genetic transformation of Impatiens to introduce novel flower colors or virus resistance has been limited by its general recalcitrance to tissue culture and transformation manipulations. We have established a regeneration and transformation system for Impatiens that provides new alternatives to genetic improvement of this crop. RESULTS: In a first step towards the development of transgenic INSV-resistant Impatiens, we developed an efficient plant regeneration system using hypocotyl segments containing cotyledonary nodes as explants. With this regeneration system, 80% of explants produced an average of 32.3 elongated shoots per initial explant plated, with up to 167 elongated shoots produced per explant. Rooting efficiency was high, and 100% of shoots produced roots within 12 days under optimal conditions, allowing plant regeneration within approximately 8 weeks. Using this regeneration system, we developed an efficient Agrobacterium-mediated Impatiens transformation method using in vitro multiple bud cultures as explants and a binary plasmid (pHB2892) bearing gfp and nptII genes. Transgenic Impatiens plants, with a frequency up to 58.9%, were obtained within 12 to 16 weeks from inoculation to transfer of transgenic plants to soil. Transgenic plants were confirmed by Southern blot, phenotypic assays and T1 segregation analysis. Transgene expression was observed in leaves, stems, roots, flowers, and fruit. The transgenic plants were fertile and phenotypically normal. CONCLUSION: We report the development of a simple and efficient Agrobacterium-mediated transformation system for Impatiens. To the best of our knowledge, there have been no reports of Agrobacterium-mediated transformation of Impatiens with experimental evidence of stable integration of T-DNA and of Agrobacterium-mediated transformation method for plants using in vitro maintained multiple bud cultures as explants. This transformation system has the advantages of 1) efficient, simple and rapid regeneration and transformation (with no need for sterilization or a greenhouse to grow stock plants), 2) flexibility (available all the time) for in vitro manipulation, 3) uniform and desirable green tissue explants for both nuclear and plastid transformation using Agrobacterium-mediated and biolistics methods, 4) no somaclonal variation and 5) resolution of necrosis of Agrobacterium-inoculated tissues.

High-efficiency transformation of the diploid strawberry (Fragaria vesca) for functional genomics.

Fragaria vesca L., a diploid (2n = 2x = 14) relative of the commercial octoploid strawberry, is an attractive model for functional genomics research in Rosaceae. Its small genome size, short reproductive cycle, and facile vegetative and seed propagation make F. vesca a promising candidate for forward and reverse genetics experiments. However, the lack of a high-efficiency transformation protocol required for systematic production of thousands of T-DNA insertional mutant lines and high-throughput gene validation is a major bottleneck. We describe a new transformation procedure that uses leaf explants from newly unfolded trifoliate leaves obtained from stock plants 6-7 weeks after seed germination, co-cultivation with Agrobacterium strain GV3101, and stringent selection on MS medium containing 4 mg l(-1) hygromycin. Using this protocol we achieved 100% transformation efficiency for 6 of 14 F. vesca accessions tested. Accession PI 551572 was determined to be the best candidate for a model in F. vesca functional genomics research, as it showed the greatest propensity for callus formation, transformation, shoot regeneration, ex vitro establishment, and plant growth, requiring only 14-15 weeks to complete its life cycle in different seasons in the greenhouse.

Age-adjusted baseline data for women with hallux valgus undergoing corrective surgery.

BACKGROUND: Functional and health-related quality-of-life data on a population of patients with hallux valgus are lacking. Similarly, the correlation of the severity of the deformity with these measures is unknown. METHODS: Two hundred and eighty-five women with an average age of forty-nine years who were scheduled for bunion surgery were enrolled in the study. The patients completed a baseline American Academy of Orthopaedic Surgeons (AAOS) foot and ankle outcomes questionnaire, which includes the Short Form-36 (SF-36) and a specific lower-extremity section on the foot and ankle. Preoperative radiographic data with regard to the hallux valgus angle and the intermetatarsal angle were stratified into groups according to the severity of the deformity (mild, moderate, or severe). The data were then stratified into age-groups consistent with those reported for the SF-36, and the results were compared with the SF-36 scores for the general population. The global foot and ankle score and the shoe comfort score were compared with general population scores that were published previously. The severity of the preoperative deformity was correlated with the baseline scores. RESULTS: General health scores were noted to be relatively stable throughout the age-groups for patients with bunions, with the older groups demonstrating better scores than the general population. Bodily pain scores were consistently worse for patients with a bunion through all age-groups compared with the general population. The average global foot and ankle score and the shoe comfort score were significantly lower (p < 0.001 for both) for the patients with a bunion than for the general population. The severity of the preoperative deformity did not correlate with any of the outcome scores. CONCLUSIONS: The bodily pain score from the SF-36 appears to be a sensitive measure of problems experienced by patients undergoing bunion surgery. Surprisingly, the severity of the deformity as measured radiographically did not correlate with any of the fifteen scores measured. These data may serve as a baseline for clinical hallux valgus studies with use of the SF-36 or the AAOS outcomes questionnaire.