Confirmation of Burkholderia gladioli as the Causal Agent of Bacterial Scab on Gladiolus (Gladiolus spp.) in Ohio.

Ohio is one of the top five floriculture producers in the United States, grossing over $200 million annually (NASS 2019). Within the international floriculture trade, gladiolus cut flowers represent the fifth highest grossing crop (Ahmed et al. 2002). In September 2021, the Ornamental Crops Pathology Lab at the Ohio State University received a gladiolus (Gladiolus spp.) sample of an unknown cultivar from a home garden in Franklin Co., OH where several plants had failed to grow from planted corms or were stunted and displaying symptoms of disease. Bleached, water-soaked spots with necrotic margins along the flowering stems, stunted flowers with partial necrosis, and necrotic bracts were observed on the submitted sample. Bacterial isolations were performed by surface disinfesting small sections of bract tissue from the border of a lesion by soaking in 10% bleach for 30 sec and rinsing twice in sterile water, macerating the tissue in sterile water, and streaking the suspension on nutrient agar (NA) plates. Plates were incubated at 28°C for 48 hours and the resulting colonies were purified by re-streaking a single colony on NA twice. Bacterial colony morphology on NA presented as cream-colored and shiny with an irregular form and undulate margin. Five in vitro tests were performed using one representative isolate to identify the bacterium to the genus level: (1) confirmed levan production, (2) confirmed pectinolytic activity, (3) confirmed ability to grow at 40°C, (4) inability to grow under anaerobic conditions, and (5) a negative oxidase test (Schaad et al. 2000). All test results identified the genus as Burkholderia. To identify to species level, gyrase subunit B (gyrB) and RNA polymerase subunit D (rpoD) markers were PCR amplified and sequenced using primers UP1-E/AprU, and 70F2/70R2, respectively (Maeda et al. 2006). NCBI GenBank BLASTn comparison showed that the gyrB sequence shared 99.33% identity to the type strain of B. gladioli (CP009323.1), while the rpoD sequence showed 99.53% identity (CP009322.1). Sequences were deposited in GenBank under accession numbers ON597852 (gyrB) and ON597853 (rpoD). To confirm pathogenicity, each of two Gladiolus communis 'Mini Elvira' potted plants were inoculated with two bacterial and two control treatments (3 leaves/treatment/plant) as follows: leaf infiltration with 1 mL of either (i) a distilled water-Tween 20 (0.03% v/v) bacterial suspension (106 cfu/mL) or (ii) a sterile water-Tween 20 suspension using a needle-less syringe; foliar spray with either (iii) the bacterial suspension or (iv) water-Tween suspension until run-off. Following inoculation, plants were covered for 24 hours with a plastic bag to increase humidity and favor infection and maintained in a greenhouse at an average temperature of 23°C. After 3 days, water-soaked, necrotic lesions were observed on the inoculated plants regardless of inoculation method, while control leaves remained asymptomatic. To fulfill Koch's postulates, bacteria were re-isolated from the lesions 7 days post-inoculation and confirmed to be identical to the original isolate based on rpoD gene sequencing. Bacterial scab of gladiolus was reported in Ohio in the late 1900s as caused by Pseudomonas gladioli (syn. P. marginata; Ellett, 1989). To the best of our knowledge, this report represents the first molecular identification of the causal agent as Burkholderia gladioli. In Ohio, the pathogen has also been observed causing slippery skin on onion but not officially reported in the peer-reviewed literature. Additionally, B. gladioli has been reported in other parts of the United States on orchid, corn, and rice (Keith et al. 2005; Lu et al. 2007; Nandakumar et al. 2009). Given the significant role of gladiolus within Ohio's floricultural trade, as well as the ability of this pathogen to infect other regional crops, monitoring of bacterial scab is important for floriculture and field crop growers alike.

First Report of Colletotrichum sansevieriae Causing Anthracnose of Snake Plant (Dracaena trifasciata) in Ohio and its Draft Genome.

Dracaena trifasciata (Prain) Mabb. is a popular houseplant in the United States. In September 2021, two diseased samples from two Ohio homeowners were received by the Ornamental Pathology Laboratory at The Ohio State University. Each sample included one or two detached leaves displaying circular gray water-soaked lesions scattered throughout the lamina and blighted areas with concentric rings bearing brown to black acervuli. Lesions covered between 25 and 50% of the leaf surface. Isolations were made by excising small portions of leaf tissue from the margin of the lesions, surface-disinfesting in 10% bleach for 45 s, rinsing in sterile water, and plating on potato dextrose agar (PDA). Plates were incubated at 23°C for one week. Two representative isolates, one per sample (FPH2021-5 and -6), were obtained by transferring hyphal tips to fresh PDA plates. Mycelia of both isolates were aerial, cottony, grayish-white, producing spores in a gelatinous orange matrix, and appeared gray to olivaceous-gray on the plate underside. Conidia produced by both isolates were cylindrical, single-celled, hyaline, measuring 12.02 to 18.11 (15.51) × 5.03 to 7.29 (6.14) µm (FPH2021-5; n=50) and 15.58 to 20.90 (18.39) × 5.63 to 8.27 (7.05) µm (FPH2021-6; n=50). Appressoria were globose to subglobose, single-celled, dark brown to sepia, measuring 6.62 to 13.98 (8.97) × 5.05 to 6.58 (6.58) µm (FPH2021-5; n=50), and 6.54 to 11.32 (8.63) × 4.54 to 8.94 (7.09) µm (FPH2021-6; n=50). Genomic DNA (gDNA) samples were extracted from both isolates and the internal transcribed spacer (ITS) region was amplified using primers ITS1F/ITS4 (Gardes and Bruns, 1993; White et al. 1990). GenBank BLAST sequence analysis resulted in 99.83% (FPH2021-5; GenBank Acc. No. OP410918.1) and 100% (FPH2021-6; OP410917.1) identity with 100% query coverage to the type strain of Colletotrichum sansevieriae Miho Nakam. & Ohzono MAFF239721 or Sa-1-2 (NR_152313.1; Nakamura et al. 2006). Whole genome sequencing was conducted for FPH2021-6 and the assembly was deposited in GenBank (JAOQIF000000000.1). The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ß-tubulin (ß-tub) regions were either extracted from the genome of FPH2021-6 (OP414603.1 and OP414601.1, respectively) or amplified from FPH2021-5 gDNA using primers GDF/GDR (OP414604.1) and Bt-2b/T1 (OP414602.1), respectively (Templeton et al. 1992; Glass and Donaldson 1995; O'Donnell and Cigelnik 1997). A multilocus partitioned analysis (Chernomor et al. 2016) based on concatenated sequences of ITS, GAPDH, and ß-tub using ModelFinder (Kalyaanamoorthy et al. 2017) was performed to build a maximum likelihood tree (IQ-TREE v2.0.3; Nguyen et al. 2015), suggesting that these two isolates are phylogenetically closer to the type strain from Japan than to a previously reported isolate 1047 from Florida (Palmateer et al. 2012). To fulfill Koch's postulates, two parallel leaf sections from one 10-inch D. trifasciata 'Laurentii' plant maintained in a 1.3-liter container were selected. Three wounds were made in each section using a sterile syringe needle. A 10-µl drop of either a 1×106 conidia/ml suspension of isolate FPH2021-6 or sterile water was placed on each wound. The plant was covered with a plastic bag for two days post-inoculation (DPI) and maintained in a greenhouse at 25°C with a 12- h photoperiod. The experiment was conducted twice. Grayish water-soaked lesions, acervuli, and leaf blight were observed on the inoculated sections 3, 10, and 14 DPI, respectively, while no symptoms appeared on the sections treated with sterile water. C. sansevieriae was re-isolated from the lesions and confirmed to be identical to the original isolate based on ITS sequencing and morphological examinations. To the best of our knowledge, this is the first report of C. sansevieriae on D. trifasciata in Ohio and the first genome draft of an isolate from the United States. Availability of whole-genome sequence data is paramount for resolving species identification in this highly diverse fungal genus, and a powerful tool to conduct comparative genomic analyses in the future.

Understanding roles in health care through interprofessional educational experiences.

BACKGROUND: Students can find interacting within a healthcare team challenging. It is important for students to understand their role and respect those of other healthcare team members. Interprofessional education (IPE) is a strategy for exploring the roles of self and others within the team. AIM: The purpose of this study was to evaluate nursing students' perceptions of roles and responsibilities following an IPE experience. METHODS: Students in an undergraduate baccalaureate degree nursing programme participated in a two-day IPE event with students in the physician's assistant's (PA) programme, pharmacy programme, and physical therapy (PT) programme. FINDINGS: Self-perception and the perception of others were two main themes that emerged. The results suggested that roles and responsibilities are often misunderstood. CONCLUSION: Educators must be committed to educating our future healthcare workforce on role expectations and responsibilities within an individual's own profession and that of others. This education should start in the foundation stages of each discipline's educational curricula.

Integration of leadership competencies in a community health simulation.

AIM: The purpose of this article is to address ways in which a nursing faculty can build nursing leadership competencies into a nursing programme. This article describes the use of simulation as a compelling learning strategy. BACKGROUND: Nurses need to be prepared to meet the demands of mass casualty events. With the growing need for major incident training, leadership competency development is a critical gap in the literature. EVALUATION: A collaborative simulation was used to help students apply classroom knowledge. In this simulation, students participated in major incident triage and the application of leadership competencies to appropriately manage victims' care. CONCLUSION: Collaboration among faculty members when designing simulation scenarios is a powerful approach to nursing education. Students were able to engage in necessary skills for their future nursing practice.

Through the lens of the American Association of Colleges of Nursing's (AACN) new Essentials: Integration of human trafficking education and simulation within baccalaureate nursing curricula.

With an increase in human trafficking in the United States, there is a need for nursing curricula to provide education and engagement in recognizing and appropriately caring for victims. The purpose of this article is to describe an undergraduate nursing simulation involving a human trafficking victim with a discussion of how the American Association of Colleges of Nursing's (AACN) Essentials were addressed by the simulation. The results of the course evaluation indicated that providing an opportunity for baccalaureate nursing students to engage in a simulation involving a victim of human trafficking allowed them to solidify classroom theory and education. Students reported increased confidence in recognizing victims after participating in the education and simulation. In addition, the simulation met many of the American Association of Colleges of Nursing's new Essentials further highlighting the importance of this clinical activity within the nursing curriculum. Nursing education is obligated to empower students to identify social determinants of health and advocate for social justice related to vulnerable populations. Because nurses are the largest group of healthcare providers, they are likely to encounter human trafficking victims and should be educated to identify victims in practice.

Utilizing Simulation-Based Training to Increase Preeclampsia Recognition in Undiagnosed Patients.

Research suggests that simulation in nursing education is a learning strategy that promotes critical thinking and utilization of clinical judgment in a safe environment without fear of retribution if errors are made. Senior nursing students in a small liberal arts university participated in a complex simulation that involved a pregnant patient with preeclampsia. The students were charged with doing a complete assessment of the patient and unborn baby. Upon completion of the assessment, the students utilized their critical thinking skills to determine the best treatment for the patient and baby that rendered the best outcomes. The goal of the exercise was to provide the students with a realistic scenario that advanced quickly into an emergent situation. The majority of students will not work in labor and delivery, but they will experience situations in their respective units that require quick thinking and critical decision-making. This simulation was a strategy to help facilitate these students into their transition to practice.

Risk factors of patients with and without postoperative nausea (PON).

This purpose of this analysis was to study risk factors of postoperative nausea (PON) and their strength. Data were obtained during the screening phase of a controlled clinical trial of aromatherapy for PON. In a sample of 1151 postsurgical subjects, 301 (26.2%) reported PON. Significant risk factors identified in the order of odds ratios for nausea were female gender, gastrointestinal surgery, use of volatile anesthesia gases, history of PON, history of motion sickness, and use of opioids after surgery. Although still over 1.0, the risk factors of length of surgery over 1 hour and gynecologic surgery had the lowest odds ratios. Likelihood of nausea increased significantly with the number of significant risk factors (P<.0001). Administration of preventive antiemetic medication also increased with the number of significant risk factors (P<.0001). Among 301 subjects reporting nausea, 49 (16.28%) received preventive medication. Despite prevention efforts, PON remains a substantial side effect for many surgical patients.

Too Sensitive or Just Right?

OBJECTIVES: To compare the performance of the rabbit monoclonal antihuman CD246 antibody (D5F3 clone) with the established ALK1 clone for immunohistochemical assessment of anaplastic large cell lymphoma (ALCL). METHODS: Archival cases of ALCL (n = 27) were assessed immunohistochemically by use of ALK1 and D5F3 clones under standard Clinical Laboratory Improvement Amendments-compliant conditions. The intensity of cytoplasmic staining (0 = none; 1 = faint; 2 = moderate; 3+ = strong) and proportion of neoplastic cells (0%, <5%, 5%-50%, >50%) were evaluated and compared with clinical ALK break-apart fluorescence in situ hybridization (FISH) assays. RESULTS: Nine ALCL specimens were positive for ALK expression by ALK1 staining (33%; 1 = 1+; 0 = 2+; 8 = 3+), while 14 were positive by D5F3 staining (48%; 3 = 1+; 1 = 2+; 10 = 3+). Across the cohort, D5F3 staining showed a significantly greater proportion of cells staining positive (P = .02) and greater intensity (P = .03). Of 3 cases positive for D5F3 only with FISH results, none showed rearrangements, although 1 showed copy number gains at the ALK locus in a subset of cells. CONCLUSIONS: Overall, D5F3 showed greater stain intensity and proportion staining than ALK1 in ALK-positive ALCL cases, which is especially helpful in limited samples. Caution and consideration of orthogonal ALK testing types is recommended, especially for cases with weak or focal staining.

Treatment of a chronically infected nasal silicone prosthesis with continuous antibiotic irrigation and gentamicin-impregnated polymethylmethacrylate beads.

Infected nasal alloplasts in revision rhinoplasty can be a complex problem, as timing between implant removal and reconstruction is the major limiting factor. Delaying reconstruction can result in loss of mechanical support, a constricted nose, and in severe cases, complete nasal airway collapse and respiratory compromise. In this case report, we describe a novel surgical approach for the management of a chronically infected nasal implant combining techniques used to treat biomaterial-associated infections: antibiotic-impregnated polymethymethacrylate beads and a continuous catheter-based antibiotic irrigation system. We report a case of a chronic alloplastic-associated infection following nasal reconstruction using a silicone implant. We utilized a two-staged approach. The involved nasal implant was removed and replaced temporarily with gentamicin-impregnated polymethymethacrylate beads and a continuous closed irrigation and drainage system with local and parenteral delivery of antibiotics. Both modalities allowed for complete eradication of the infection. In addition, the gentamicin beads provided sufficient mechanical support in order to minimize the risk of skin contracture. Twelve days after her initial surgery, nasal reconstruction was performed using a cadaver bone graft. The patient was followed for two years postoperatively and has shown good results with no evidence of skin contracture or recurrent infection. This technique may allow for shorter delay in revision surgery and reduce the risk of long-term complications without compromising functional and aesthetic outcomes.

Repeated aerosolized AAV-CFTR for treatment of cystic fibrosis: a randomized placebo-controlled phase 2B trial.

Previous studies have demonstrated that delivery of a recombinant adeno-associated virus (AAV) vector encoding the complete human cystic fibrosis transmembrane regulator (CFTR) cDNA (tgAAVCF) to the nose, sinus, and lungs of subjects with cystic fibrosis (CF) was safe and well tolerated. In a small randomized, double-blind study of three doses of aerosolized tgAAVCF or placebo at 30-day intervals, encouraging but non-significant trends in pulmonary function and induced sputum interleukin 8 (IL-8) levels were seen at early time points. This larger study was conducted to verify these trends. One hundred and two subjects aged 12 years and older with mild-to-moderate cystic fibrosis (forced expiratory flow in 1 sec [FEV1]:60% predicted) were randomized to two aerosolized doses of 1x10(13)DNase-resistant particles of tgAAVCF (n=51) or matching placebo (n=51) administered 30 days apart. Although tgAAVCF was well tolerated, the study did not meet its primary efficacy end point of statistically significant improvement in FEV1 30 days after initial administration of tgAAVCF compared with placebo. There were no significant differences in spirometric lung function over time, induced sputum biologic markers, or days of antibiotic use in either treatment group. Thus repeated doses of aerosolized tgAAVCF were safe and well tolerated, but did not result in significant improvement in lung function over time. Because gene transfer is the simplest, most basic way to correct the underlying genetic defect that leads to disease in CF, further research is warranted to develop an effective gene transfer agent for the treatment of CF.

QSPR modeling of solubility of polyaromatic hydrocarbons and fullerene in 1-octanol and n-heptane.

Solubility of polyaromatic hydrocarbons (PAH) and carbon nanostructures is important both from the technical and environmental points of view. In the present work, two general quantitative structure-property relationship (QSPR) models were developed, describing the solubility of PAH-s and fullerene (C60) in two different condensed media (1-octanol and n-heptane). Statistically good QSPR models were obtained by using forward selection techniques from large space of theoretical molecular descriptors. The physical meaning of the models is discussed and analyzed.

Duchenne Regulatory Science Consortium Meeting on Disease Progression Modeling for Duchenne Muscular Dystrophy.

INTRODUCTION: The Duchenne Regulatory Science Consortium (D-RSC) was established to develop tools to accelerate drug development for DMD.  The resulting tools are anticipated to meet validity requirements outlined by qualification/endorsement pathways at both the U.S. Food and Drug Administration (FDA) and European Medicines Administration (EMA), and will be made available to the drug development community. The initial goals of the consortium include the development of a disease progression model, with the goal of creating a model that would be used to forecast changes in clinically meaningful endpoints, which would inform clinical trial protocol development and data analysis.  Methods: In April of 2016 the consortium and other experts met to formulate plans for the development of the model.  Conclusions: Here we report the results of the meeting, and discussion as to the form of the model that we plan to move forward to develop, after input from the regulatory authorities.

Effect of adeno-associated virus-specific immunoglobulin G in human amniotic fluid on gene transfer.

Intra-amniotic administration of adeno-associated virus (AAV) vector may be an effective way to deliver gene therapy for treatment of congenital pulmonary and intestinal disorders. In an effort to understand potential barriers to intra-amniotic gene therapy better, we determined whether human amniotic fluid (AF) could act as an inhibitor of AAV2-mediated gene transfer. AF samples were obtained from 21 different human pregnancies during routine amniocentesis at 16-20 weeks of gestation. An immortalized fetal human tracheal epithelial cell line (FHTE) was infected with AAV2 containing a luciferase reporter gene driven by the SV40 promoter in the presence and absence of each AF sample. Inhibition of transgene expression was observed in 8 (38%) of the AF samples (inhibitory AF) and resulted in luciferase levels of only 1.4% +/- 0.6% of those obtained with infection in normal media. Infections in 13 samples (62%) resulted in transgene expression comparable or in excess of infection in media alone (noninhibitory AF). Removal of immunoglobulin G (IgG) from inhibitory AF samples with Protein A returned luciferase expression to control levels (119% +/- 37% of control), suggesting the possible presence of inhibiting antibody. Eleven of the AF samples were evaluated by enzyme-linked immunosorbent assay (ELISA) for specific anti-AAV antibodies. All noninhibitory AF samples were negative (titers of < 1:20; n = 3), and 6 of the 8 inhibitory samples contained specific anti-AAV antibodies at titers ranging from 1:40 to 1:160. These studies demonstrate that AF from some individuals contains AAV-specific IgG that can inhibit gene transfer.

Hiring relatives as caregivers in two states: developing an education and research agenda for policy makers.

Meeting the long-term care needs of the growing aging population is a priority policy issue in the United States. Yet, hiring relatives as caregivers remains a controversial policy issue. This two-state case study reports findings about views from policy experts regarding a policy option to hire family caregivers in home- and community-based long-term care programs. Policy makers also discussed information needed by other states considering this option and effective approaches to disseminate findings about this option from the Cash and Counseling Demonstration Evaluation. Based on the study results, we present an education and research agenda to communicate these findings to policy makers.

Evaluation of exposure and health care worker response to nebulized administration of tgAAVCF to patients with cystic fibrosis.

A study was conducted to assess health care worker exposure to tgAAVCF during the aerosolized administration of this experimental gene transfer agent in clinical trials for the treatment of cystic fibrosis (CF). tgAAVCF is a recombinant adeno-associated virus (AAV) genetically engineered to contain the human CF transmembrane conductance regulator cDNA. Study subjects included eight health care workers involved in the administration of tgAAVCF in a phase II study and 12 control health care workers who were involved with the treatment of CF patients, but not administration of the study drug. The exposure assessment entailed the determination of personal and area airborne tgAAVCF concentrations. In addition, serologic status of the health care workers was evaluated throughout the study for the presence of antibodies to AAV. A symptom survey was also completed by both the active and control health care workers. Air samples were analyzed by an infectivity assay (active vector) and a DNA polymerase chain reaction amplification procedure (vector DNA). Air monitoring was conducted during 13 tgAAVCF and seven placebo administrations. Active vector and vector particles were detected in four of 51 and 48 of 51 air samples collected during the administration of tgAAVCF, respectively. Based on the airborne vector particle concentration, the workers' exposure was estimated to be 0.0006% of the administered dose. At this level of exposure, the prevalence of symptoms was very low, the spectrum was similar in both study groups and did not result in any reported negative health effects.