Evaluation of changes in knowledge and attitude among youth after a one-hour introduction to workplace safety and health: Safety Matters.

INTRODUCTION: Young workers in the United States are injured at higher rates than adults, a trend that has persisted for more than two decades. Despite known risks, young people enter the workforce with little-or-no preparation for the hazards they may face. In 2016, the National Institute for Occupational Safety and Health and American Industrial Hygiene Association developed Safety Matters, a one-hour educational module to raise awareness of workplace safety and health among young people. METHOD: A pilot project was conducted to evaluate the effectiveness of Safety Matters to positively change workplace safety and health knowledge and attitude scores among a sample of 283 youth in Colorado. Train-the-trainer sessions prepared volunteer safety and health professionals to deliver Safety Matters with fidelity and to conduct the assessment immediately prior to and following the program. RESULTS: After receiving Safety Matters, participants had statistically significant (p < 0.001) increased scores for both workplace safety and health knowledge (Cohen's d = 1.12; large effect size) and importance (attitude) (Cohen's d = 0.51; medium effect size). Although univariate analyses showed knowledge and attitude scores significantly increased for all demographic groups examined, there were statistically significant differences in knowledge scores by participant age (p < 0.01), ethnicity (p < 0.05), and race (p < 0.001) and statistically significant differences in attitude scores by participant race (p < 0.001). However, when race and ethnicity were both used as predictors in a regression model, only race continued to predict statistically significant (p < 0.01) changes in knowledge and attitude. CONCLUSIONS: This project introduces a promising, community-based model for a one-hour introduction to workplace safety and health on which future, job-specific safety training can be built. PRACTICAL APPLICATIONS: Safety and health professionals can play a critical role in promoting the health and safety of young workers. Adapting health and safety programs to diverse youth populations may enhance program relevance and receptivity.

The Combination of Trametinib and Ganitumab is Effective in RAS-Mutated PAX-Fusion Negative Rhabdomyosarcoma Models.

PURPOSE: PAX-fusion negative rhabdomyosarcoma (FN RMS) is driven by alterations in the RAS/MAP kinase pathway and is partially responsive to MEK inhibition. Overexpression of IGF1R and its ligands is also observed in FN RMS. Preclinical and clinical studies have suggested that IGF1R is itself an important target in FN RMS. Our previous studies revealed preclinical efficacy of the MEK1/2 inhibitor, trametinib, and an IGF1R inhibitor, BMS-754807, but this combination was not pursued clinically due to intolerability in preclinical murine models. Here, we sought to identify a combination of an MEK1/2 inhibitor and IGF1R inhibitor, which would be tolerated in murine models and effective in both cell line and patient-derived xenograft models of RAS-mutant FN RMS. EXPERIMENTAL DESIGN: Using proliferation and apoptosis assays, we studied the factorial effects of trametinib and ganitumab (AMG 479), a mAb with specificity for human and murine IGF1R, in a panel of RAS-mutant FN RMS cell lines. The molecular mechanism of the observed synergy was determined using conventional and capillary immunoassays. The efficacy and tolerability of trametinib/ganitumab was assessed using a panel of RAS-mutated cell-line and patient-derived RMS xenograft models. RESULTS: Treatment with trametinib and ganitumab resulted in synergistic cellular growth inhibition in all cell lines tested and inhibition of tumor growth in four of six models of RAS-mutant RMS. The combination had little effect on body weight and did not produce thrombocytopenia, neutropenia, or hyperinsulinemia in tumor-bearing SCID beige mice. Mechanistically, ganitumab treatment prevented the phosphorylation of AKT induced by MEK inhibition alone. Therapeutic response to the combination was observed in models without a mutation in the PI3K/PTEN axis. CONCLUSIONS: We demonstrate that combined trametinib and ganitumab is effective in a genomically diverse panel of RAS-mutated FN RMS preclinical models. Our data also show that the trametinib/ganitumab combination likely has a favorable tolerability profile. These data support testing this combination in a phase I/II clinical trial for pediatric patients with relapsed or refractory RAS-mutated FN RMS.

Evaluation of an Occupational Safety and Health Training for Cannabis Cultivation Workers.

OBJECTIVES: As the commercial cannabis industry grows, there is an increased need to characterize potentially hazardous workplace exposures and provide training to workers to mitigate these exposures with the goal of reducing accidents and injuries from cannabis cultivation, processing, and manufacturing. Public health and safety stakeholders in Colorado developed a worker-focused training designed to improve hazard awareness, recognition, and controls related to commercial cannabis cultivation. This paper describes the evaluation of this training. METHODS: The training was a full day, in-person educational experience directed to workers in the cannabis cultivation industry. Training topics included an overview of occupational safety and health hazards, chemical exposures, slip, trips, and falls, repetitive motion, the application of the hierarchy of control including lockout/tagout, machine guarding, personal protective equipment, among others. Evaluation surveys assessed attendee demographics, perceived job hazards, confidence to change workplace practices, knowledge, training relevancy and quality, intent to change behavior, as well as barriers and resources. RESULTS: A total of 208 people attended the safety trainings. One hundred and thirty-four participants (64%) completed the pre-training survey and 107 (51%) completed the post-training survey. Respondents provided high ratings for the quality and relevance of the training, with 91.3% of respondents rating the training very good or excellent. Before the training, the attendees listed their most concerning safety and health issues as exposure to pesticides and other chemicals (65.7%), absorbing chemicals through the skin (56.7%), slips, trips, and falls (52.2%), and respiratory hazards (50.7%). After the training, they reported the most concerning hazards to be slips, trips, and fall hazards (65.4%), ergonomic problems (64.5%), and respiratory issues (61.7%). There was a statistically non-significant increase in knowledge scores from 67.1% correct to 76.0% correct. Finally, 88.5% of respondents felt extremely or very confident that they could change their own health and safety practices at work. CONCLUSIONS: The training successfully reached cannabis employees in cultivation, compliance, and management. Survey respondents felt that the training was of high quality and addressed gaps in their knowledge related to safety and health hazards in the cannabis industry. The workplace safety and health concerns shifted from pre- to post-training. There was a statistically non-significant increase in knowledge. Additional follow-up of training attendees would be beneficial to measure sustained impact of training.

Eosinophils Do Not Drive Acute Muscle Pathology in the mdx Mouse Model of Duchenne Muscular Dystrophy.

Eosinophils are present in muscle lesions associated with Duchenne muscular dystrophy and dystrophin-deficient mdx mice that phenocopy this disorder. Although it has been hypothesized that eosinophils promote characteristic inflammatory muscle damage, this has not been fully examined. In this study, we generated mice with the dystrophin mutation introduced into PHIL, a strain with a transgene that directs lineage-specific eosinophil ablation. We also explored the impact of eosinophil overabundance on dystrophinopathy by introducing the dystrophin mutation into IL-5 transgenic mice. We evaluated the degree of eosinophil infiltration in association with myofiber size distribution, centralized nuclei, serum creatine kinase, and quantitative histopathology scores. Among our findings, eosinophils were prominent in the quadriceps muscles of 4-wk-old male mdx mice but no profound differences were observed in the quantitative measures of muscle damage when comparing mdx versus mdx.PHIL versus mdx.IL5tg mice, despite dramatic differences in eosinophil infiltration (CD45+CD11c-Gr1-MHC class IIloSiglecF+ eosinophils at 1.2 ± 0.34% versus <0.1% versus 20 ± 7.6% of total cells, respectively). Further evaluation revealed elevated levels of eosinophil chemoatttractants eotaxin-1 and RANTES in the muscle tissue of all three dystrophin-deficient strains; eotaxin-1 concentration in muscle correlated inversely with age. Cytokines IL-4 and IL-1R antagonist were also detected in association with eosinophils in muscle. Taken together, our findings challenge the long-held perception of eosinophils as cytotoxic in dystrophin-deficient muscle; we show clearly that eosinophil infiltration is not a driving force behind acute muscle damage in the mdx mouse strain. Ongoing studies will focus on the functional properties of eosinophils in this unique microenvironment.

Current Marijuana Use by Industry and Occupation - Colorado, 2014-2015.

The effects of marijuana use on workplace safety are of concern for public health and workplace safety professionals. Twenty-nine states and the District of Columbia have enacted laws legalizing marijuana at the state level for recreational and/or medical purposes. Employers and safety professionals in states where marijuana use is legal have expressed concerns about potential increases in occupational injuries, such as on-the-job motor vehicle crashes, related to employee impairment. Data published in 2017 by the Colorado Department of Public Health and Environment (CDPHE) showed that more than one in eight adult state residents aged ≥18 years currently used marijuana in 2014 (13.6%) and 2015 (13.4%) (1). To examine current marijuana use by working adults and the industries and occupations in which they are employed, CDPHE analyzed data from the state's Behavioral Risk Factor Surveillance System (BRFSS) regarding current marijuana use (at least 1 day during the preceding 30 days) among 10,169 persons who responded to the current marijuana use question. During 2014 and 2015, 14.6% of these 10,169 Colorado workers reported current marijuana use, with the highest reported prevalence among workers in the Accommodation and Food Services industry (30.1%) and Food Preparation and Serving (32.2%) occupations. Understanding the industries and occupations of adults with reported marijuana use can help direct and maximize impact of public health messaging and potential safety interventions for adults.

Splenogonadal fusion: a rare etiology of solid testicular mass.

A 35-year-old man presented with a painless left scrotal mass. Pathologic examination after orchiectomy revealed splenogonadal fusion. Splenogonadal fusion is an exceptionally rare, typically benign, congenital anomaly. Splenogonadal fusion should be included in the differential diagnosis of a left-sided testicular mass.

Optimizing Frozen Sample Preparation for Laser Microdissection: Assessment of CryoJane Tape-Transfer System®.

Laser microdissection is an invaluable tool in medical research that facilitates collecting specific cell populations for molecular analysis. Diversity of research targets (e.g., cancerous and precancerous lesions in clinical and animal research, cell pellets, rodent embryos, etc.) and varied scientific objectives, however, present challenges toward establishing standard laser microdissection protocols. Sample preparation is crucial for quality RNA, DNA and protein retrieval, where it often determines the feasibility of a laser microdissection project. The majority of microdissection studies in clinical and animal model research are conducted on frozen tissues containing native nucleic acids, unmodified by fixation. However, the variable morphological quality of frozen sections from tissues containing fat, collagen or delicate cell structures can limit or prevent successful harvest of the desired cell population via laser dissection. The CryoJane Tape-Transfer System®, a commercial device that improves cryosectioning outcomes on glass slides has been reported superior for slide preparation and isolation of high quality osteocyte RNA (frozen bone) during laser dissection. Considering the reported advantages of CryoJane for laser dissection on glass slides, we asked whether the system could also work with the plastic membrane slides used by UV laser based microdissection instruments, as these are better suited for collection of larger target areas. In an attempt to optimize laser microdissection slide preparation for tissues of different RNA stability and cryosectioning difficulty, we evaluated the CryoJane system for use with both glass (laser capture microdissection) and membrane (laser cutting microdissection) slides. We have established a sample preparation protocol for glass and membrane slides including manual coating of membrane slides with CryoJane solutions, cryosectioning, slide staining and dissection procedure, lysis and RNA extraction that facilitated efficient dissection and high quality RNA retrieval from CryoJane preparations. CryoJane technology therefore has the potential to facilitate standardization of laser microdissection slide preparation from frozen tissues.

Tissue preparation for MALDI-MS imaging of protein and peptides.

Matrix-assisted laser desorption/ionization (MALDI) imaging is rapidly gaining importance in the -biomarkers field because it is able to detect several analytes at the same time and to assign to each one of them not only an m/z value but also spatial distribution. Here we present the detailed description of sample preparation for protein and peptide MALDI imaging assays. This chapter describes the microtomy performed in a cryostat to produce tissue slides mounted onto glass slides suitable for MALDI. Sample preparation will include matrix coating procedure with a sensor-controlled aerosol. Finally, we will show some examples of how data can be visualized to suit the purposes of the research.

Increased calcium levels and prolonged shelf life in tomatoes expressing Arabidopsis H+/Ca2+ transporters.

Here we demonstrate that fruit from tomato (Lycopersicon esculentum) plants expressing Arabidopsis (Arabidopsis thaliana) H(+)/cation exchangers (CAX) have more calcium (Ca2+) and prolonged shelf life when compared to controls. Previously, using the prototypical CAX1, it has been demonstrated that, in yeast (Saccharomyces cerevisiae) cells, CAX transporters are activated when the N-terminal autoinhibitory region is deleted, to give an N-terminally truncated CAX (sCAX), or altered through specific manipulations. To continue to understand the diversity of CAX function, we used yeast assays to characterize the putative transport properties of CAX4 and N-terminal variants of CAX4. CAX4 variants can suppress the Ca2+ hypersensitive yeast phenotypes and also appear to be more specific Ca2+ transporters than sCAX1. We then compared the phenotypes of sCAX1- and CAX4-expressing tomato lines. The sCAX1-expressing tomato lines demonstrate increased vacuolar H(+)/Ca2+ transport, when measured in root tissue, elevated fruit Ca2+ level, and prolonged shelf life but have severe alterations in plant development and morphology, including increased incidence of blossom-end rot. The CAX4-expressing plants demonstrate more modest increases in Ca2+ levels and shelf life but no deleterious effects on plant growth. These findings suggest that CAX expression may fortify plants with Ca2+ and may serve as an alternative to the application of CaCl2 used to extend the shelf life of numerous agriculturally important commodities. However, judicious regulation of CAX transport is required to assure optimal plant growth.

Genetic manipulation for enhancing calcium content in potato tuber.

Increased calcium (Ca) in potatoes may increase the production rate by enhancing tuber quality and storability. Additionally, increased Ca levels in important agricultural crops may help ameliorate the incidence of osteoporosis. However, the capacity to alter Ca levels in potato tubers through genetic manipulations has not been previously addressed. Here we demonstrate that potato tubers expressing the Arabidopsis H+/Ca2+ transporter sCAX1 (N-terminal autoinhibitory domain truncated version of CAtion eXchanger 1) contain up to 3-fold more Ca than wild-type tubers. The increased Ca appears to be distributed throughout the tuber. The sCAX1-expressing potatoes have normally undergone the tuber/plant/tuber cycle for three generations; the trait appeared stable through successive generations. The expression of sCAX1 does not appear to alter potato growth and development. Furthermore, increased Ca levels in sCAX1-expressing tubers do not appear to alter tuber morphology or yield. Given the preponderance of potato consumption worldwide, these transgenic plants may be a means of marginally increasing Ca intake levels in the population. To our knowledge, this study represents the first attempts to use biotechnology to increase the Ca content of potatoes.

Efficient and genotype-independent Agrobacterium--mediated tomato transformation.

An efficient method to transform five cultivars of tomato (Lycopersicon esculentum), Micro-Tom, Red Cherry, Rubion, Piedmont, and E6203 is reported. A comparison was made of leaf, cotyledon, and hypocotyl explants on 7 different regeneration media without Agrobacterium tumefaciens cocultivation and on 11 different media with cocultivation. Although all cultivars and explants formed callus and regenerated on the initial 7 media, cocultivation with A. tumefaciens significantly reduced the callus induction and regeneration. From these experiments, a transformation methodology using either hypocotyls or cotyledons cultured for one day on BA 1 mgL-1, NAA 0.1 mgL-1 and 3 days cocultivation with the Agrobacterium on this same medium followed by a transfer to a medium with zeatin 2 mgL-1 and IAA 0.1 mgL-1 for 4-6 weeks resulted in a greater than 20% transformation frequency for all five cultivars tested. In this transformation method, no feeder layers of tobacco, petunia or tomato suspension cultures were used, and the subculture media was minimal. Stable integration and transmission of the transgene in T1 generation plants were confirmed by Southern blot analysis. This procedure represents a simple, efficient and general means of transforming tomato.

How accurate is frozen section for skin cancers?

Frozen section analysis has been used to increase the likelihood of complete excision of skin cancers and to minimize the risk for recurrence. The question of its accuracy has been addressed in many studies and this article adds data to the discussion. A retrospective study was performed of 60 consecutive cases in which frozen section diagnoses were compared with permanent sections. It was found that in 85% of the cases the frozen sections were accurate compared with the permanent sections, but in 13% of the total cases the margins were less than 1 mm. These results concur with the literature. The surgery and pathological examinations were performed in a community hospital in which there was no dermatopathologist. The authors review the limitations of frozen section diagnosis in skin cancers and show that frozen section may be necessary in select cases but not in most skin cancers.

An evaluation of the Man to Man self-help group in Colorado and Utah.

PURPOSE: Two surveys were conducted to evaluate the Man to Man program, a prostate cancer self-help group of the American Cancer Society (ACS). DESCRIPTION OF STUDY: The study involved two surveys in Colorado and Utah in 2001, one of program participants, the other of area urologists, respectively. A focus group also was conducted of African American men with prostate cancer in Denver, Colo. RESULTS: In total, 112 participants in the Man to Man program and 66 urologists participated in the study. Results of the participant survey indicated that men and their families found out about the group through other prostate cancer survivors, doctors, or the ACS. Monthly attendance at the groups ranged from 2 to 50 participants. Participants valued the prostate cancer treatment information, physician presentations, and social aspects that were offered. The results of the urologist survey indicated that 67% were aware of the Man to Man program, but most were not aware of the services it offered. Urologists learned of the group through Man to Man participants, other physicians, and the ACS. Focus group results suggested that a personal preference for access to other sources of support and information was a primary reason for nonattendance at Man to Man meetings. CLINICAL IMPLICATIONS: Attendance at Man to Man meetings is likely to be enhanced by focusing on basic prostate cancer information topics. Future evaluations of Man to Man meeting participation should focus on the role of women in the program. Most urologists were not familiar with the services offered by Man to Man. The ACS and the organizers of local Man to Man programs should target urologists in future outreach efforts.