Comparative study of Rapid DNA versus conventional methods on compromised bones.

Equivalent amounts of compromised bones were used to directly compare STR success of conventional and Rapid DNA methods. Conventional DNA extraction methods, including manual full demineralization and semi-automated PrepFiler BTA/ AutoMate Express (ThermoFisher Scientific), provided insights regarding the DNA quantity and extent of degradation of each compromised bone analyzed with ANDE 6C (ANDE Corp) and RapidHIT ID (ThermoFisher Scientific) Rapid systems. Full demineralization provided higher DNA yields than extraction with the AutoMate Express for quality control (QC) and environmentally challenged bones. The degradation indices ranged from ∼1.8 to 73. Both demineralization and AutoMate Express extracts benefited from additional clean-up with NucleoSpin XS devices, which usually resulted in more alleles being detected than without further clean-up. Complete "CODIS 20″ profiles could be obtained with bone QC1 with all methods. However, among the 14 compromised bones with low DNA content, complete CODIS 20 profiles were detected for 7, 4, and 0 bones analyzed with demineralization, AutoMate Express and ANDE methods, respectively. The RapidHIT ID was the least sensitive method, providing the fewest detectable alleles for the bones tested. Whereas extracted DNA of approximately 0.1 ng can yield complete GlobalFiler STR profiles, at least 30 ng was required for complete FlexPlex 27 profiles using the ANDE 6C Rapid DNA system. In addition to being less sensitive than conventional methods, the tested Rapid DNA approaches were less predictable when attempting to improve STR success and proved to be less reliable in genotyping accuracy.

Music Upper Limb Therapy-Integrated Provides a Feasible Enriched Environment and Reduces Post-stroke Depression: A Pilot Randomized Controlled Trial.

OBJECTIVE: This study's aims were to refine Music Upper Limb Therapy-Integrated (MULT-I) to create a feasible enriched environment for stroke rehabilitation and compare its biologic and behavioral effects with that of a home exercise program (HEP). DESIGN: This was a randomized mixed-methods study of 30 adults with post-stroke hemiparesis. Serum brain-derived neurotrophic factor and oxytocin levels measured biologic effects, and upper limb function, disability, quality of life, and emotional well-being were assessed as behavioral outcomes. Participant experiences were explored using semistructured interviews. RESULTS: MULT-I participants showed reduced depression from preintervention to postintervention as compared with HEP participants. Brain-derived neurotrophic factor levels significantly increased for MULT-I participants but decreased for HEP participants, with a significant difference between groups after excluding those with post-stroke depression. MULT-I participants additionally improved quality of life and self-perceived physical strength, mobility, activity, participation, and recovery from preintervention to postintervention. HEP participants improved upper limb function. Qualitatively, MULT-I provided psychosocial support and enjoyment, whereas HEP supported self-management of rehabilitation. CONCLUSIONS: Implementation of a music-enriched environment is feasible, reduces post-stroke depression, and may enhance the neural environment for recovery via increases in brain-derived neurotrophic factor levels. Self-management of rehabilitation through an HEP may further improve upper limb function.

Pre- and Early Post-treatment With Arthrospira platensis (Spirulina) Extract Impedes Lipopolysaccharide-triggered Neuroinflammation in Microglia.

Background: Uncontrolled neuroinflammation and microglia activation lead to cellular and tissue damage contributing to neurodegenerative and neurological disorders. Spirulina (Arthrospira platensis (Nordstedt) Gomont, or Spirulina platensis), a blue-green microalga, which belongs to the class of cyanobacteria, has been studied for its numerous health benefits, which include anti-inflammatory properties, among others. Furthermore, in vivo studies have highlighted neuroprotective effects of Spirulina from neuroinflammatory insults in different brain areas. However, the mechanisms underlying the anti-inflammatory effect of the microalga are not completely understood. In this study we examined the effect of pre- and post-treatment with an acetone extract of Spirulina (E1) in an in vitro model of LPS-induced microglia activation. Methods: The effect of E1 on the release of IL-1ß and TNF-α, expression of iNOS, nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1), and the activation of NF-κB was investigated in primary microglia by ELISA, real-time PCR, and immunofluorescence. Results: Pre- and early post-treatment with non-cytotoxic concentrations of E1 down-regulated the release of IL-1ß and TNF-α, and the over-expression of iNOS induced by LPS. E1 also significantly blocked the LPS-induced nuclear translocation of NF-κB p65 subunit, and upregulated gene and protein levels of Nrf2, as well as gene expression of HO-1. Conclusions: These results indicate that the extract of Spirulina can be useful in the control of microglia activation and neuroinflammatory processes. This evidence can support future in vivo studies to test pre- and post-treatment effects of the acetone extract from Spirulina.

Mitochondrial Sequencing of Missing Persons DNA Casework by Implementing Thermo Fisher's Precision ID mtDNA Whole Genome Assay.

The advent of massively parallel sequencing (MPS) in the past decade has opened the doors to mitochondrial whole-genome sequencing. Mitochondrial (mt) DNA is used in forensics due to its high copy number per cell and maternal mode of inheritance. Consequently, we have implemented the Thermo Fisher Precision ID mtDNA Whole Genome panel coupled with the Ion Chef™ and Ion S5™ for MPS analysis in the California Department of Justice, Missing Persons DNA Program. Thirty-one mostly challenging samples (degraded, inhibited, low template, or mixed) were evaluated for this study. The majority of these samples generated single source full or partial genome sequences with MPS, providing information in cases where previously there was none. The quantitative and sensitive nature of MPS analysis was beneficial, but also led to detection of low-level contaminants. In addition, we found Precision ID to be more susceptible to inhibition than our legacy Sanger assay. Overall, the success rate (full single source hypervariable regions I and II (HVI/HVII) for Sanger and control region for MPS result) for these challenging samples increased from 32.3% with Sanger sequencing to 74.2% with the Precision ID assay. Considering the increase in success rate, the simple workflow and the higher discriminating potential of whole genome data, the Precision ID platform is a significant improvement for the CA Department of Justice Missing Persons DNA Program.

Examination of Adsorption Orientation of Amyloidogenic Peptides Over Nano-Gold Colloidal Particle Surfaces.

The adsorption of amyloidogenic peptides, amyloid beta 1-40 (Aß1-40), alpha-synuclein (α-syn), and beta 2 microglobulin (ß2m), was attempted over the surface of nano-gold colloidal particles, ranging from d = 10 to 100 nm in diameter (d). The spectroscopic inspection between pH 2 and pH 12 successfully extracted the critical pH point (pHo) at which the color change of the amyloidogenic peptide-coated nano-gold colloids occurred due to aggregation of the nano-gold colloids. The change in surface property caused by the degree of peptide coverage was hypothesized to reflect the ΔpHo, which is the difference in pHo between bare gold colloids and peptide coated gold colloids. The coverage ratio (Θ) for all amyloidogenic peptides over gold colloid of different sizes was extracted by assuming Θ = 0 at ΔpHo = 0. Remarkably, Θ was found to have a nano-gold colloidal size dependence, however, this nano-size dependence was not simply correlated with d. The geometric analysis and simulation of reproducing Θ was conducted by assuming a prolate shape of all amyloidogenic peptides. The simulation concluded that a spiking-out orientation of a prolate was required in order to reproduce the extracted Θ. The involvement of a secondary layer was suggested; this secondary layer was considered to be due to the networking of the peptides. An extracted average distance of networking between adjacent gold colloids supports the binding of peptides as if they are "entangled" and enclosed in an interfacial distance that was found to be approximately 2 nm. The complex nano-size dependence of Θ was explained by available spacing between adjacent prolates. When the secondary layer was formed, Aß1-40 and α-syn possessed a higher affinity to a partially negative nano-gold colloidal surface. However, ß2m peptides tend to interact with each other. This difference was explained by the difference in partial charge distribution over a monomer. Both Aß1-40 and α-syn are considered to have a partial charge (especially δ+) distribution centering around the prolate axis. The ß2m, however, possesses a distorted charge distribution. For a lower Θ (i.e., Θ <0.5), a prolate was assumed to conduct a gyration motion, maintaining the spiking-out orientation to fill in the unoccupied space with a tilting angle ranging between 5° and 58° depending on the nano-scale and peptide coated to the gold colloid.

Design and short-term impact of an event to promote careers in clinical pharmacy.

BACKGROUND AND PURPOSE: There is a role for local pharmacy organizations to promote clinical pharmacy and increase awareness to both potential and current pharmacy students. The Greater Milwaukee College of Clinical Pharmacy (GMCCP) chapter sought to promote clinical pharmacy amongst current and prospective pharmacy students to increase the knowledge, awareness, and interest in clinical pharmacy. EDUCATIONAL ACTIVITY AND SETTING: Subcommittee members designed programming to introduce the basics of clinical pharmacy. Students from three schools of pharmacy and more than 40 colleges in the region were invited to the event. Didactic and discussion-based content was developed. Pharmacists from GMCCP were solicited as presenters, along with a current pharmacy student and resident. Participants were asked to complete pre-event and post-event surveys to assess their awareness of and interest in clinical pharmacy. Volunteer pharmacists were also surveyed after the event. FINDINGS: Twenty-eight individuals attended the event. Nineteen students completed both the pre- and post-survey. The comparison between median score pre- and post-event was analyzed with the Wilcoxon Signed-Rank test. Students' awareness and knowledge of clinical pharmacy improved after attending the event. Students' interest in a career in clinical pharmacy also increased after attending the event. Nine of the thirteen volunteer breakout session pharmacists (69%) responded to the post-event survey. The majority strongly agreed that participants were interested in learning about their area of practice and asked meaningful questions appropriate for their area of practice. DISCUSSION AND CONCLUSIONS: The Exploring Careers in Clinical Pharmacy event increased awareness of clinical pharmacy career choices available within the profession of pharmacy. Local, regional, and national pharmacy organizations may consider utilizing the diverse talent of their membership and collaborate with colleges of pharmacy or other organizations to impact the knowledge, awareness, and interest in clinical pharmacy as a future career choice for pre-pharmacy and current pharmacy students.

A genetic basis for the variation in the vulnerability of cancer to DNA damage.

Radiotherapy is not currently informed by the genetic composition of an individual patient's tumour. To identify genetic features regulating survival after DNA damage, here we conduct large-scale profiling of cellular survival after exposure to radiation in a diverse collection of 533 genetically annotated human tumour cell lines. We show that sensitivity to radiation is characterized by significant variation across and within lineages. We combine results from our platform with genomic features to identify parameters that predict radiation sensitivity. We identify somatic copy number alterations, gene mutations and the basal expression of individual genes and gene sets that correlate with the radiation survival, revealing new insights into the genetic basis of tumour cellular response to DNA damage. These results demonstrate the diversity of tumour cellular response to ionizing radiation and establish multiple lines of evidence that new genetic features regulating cellular response after DNA damage can be identified.

Evaluation of a Patient Perspective Module in a Required Medication Safety and Quality Course at a College of Pharmacy.

OBJECTIVES: To develop and evaluate the impact of a module discussing the patient's perspective on medication errors in a required medication safety course at a college of pharmacy. METHODS: Students were required to read Josie's Story, a true story written by a mother after her daughter died from a medical error, and attend an in-class discussion regarding the book. A questionnaire, which employed a pre-post retrospective method and extracted items from the Caring Ability Inventory, was then administered to measure the change in students' perceptions of patient care. Additional questions gathered students' perceptions of the assignment, their personal experiences with the topic, and the importance of medication safety. RESULTS: 120 out of 138 students (response rate = 87%) completed the questionnaire. 56% of students indicated they would be more likely to speak with a patient directly about a medication error after reading the book, whereas only 3% were less likely, and 42% indicated they were just as likely. Most students (59%) reported that they felt more motivated to learn about medication safety after reading Josie's Story. IMPLICATIONS: This course previously addressed strategies to prevent medication errors. Successfully adding a component that introduces how a medication error impacted a patient and her family may help motivate students to recognize the importance and need for a culture of safety, personalize how medication errors impact patients, and provide a venue for students to gain patient centeredness and caring skills.

An online virtual-patient program to teach pharmacists and pharmacy students how to provide diabetes-specific medication therapy management.

OBJECTIVE: To develop, implement, and assess the effectiveness of an online medication therapy management (MTM) program to train pharmacists and pharmacy students in providing MTM services for patients with diabetes and to increase their intent to perform these services. DESIGN: An online program was created using an Internet-based learning platform to simulate 4 MTM meetings between a pharmacist and a virtual patient diagnosed with diabetes. ASSESSMENT: Eighty students and 42 pharmacists completed the program. After completing the program, scores on post-intervention assessments showed significant improvement in 2 areas: control over performing MTM, and knowledge of how to perform MTM. Students had a significantly less-positive attitude about MTM and a decline in their perception of the social expectation that MTM is part of the practice of pharmacy, while pharmacists' attitudes did not change significantly in these areas. CONCLUSION: This online program using a virtual patient improved both participants' belief that they have control over performing MTM, and their knowledge of how to perform MTM for diabetic patients, which may increase the likelihood that pharmacists and pharmacy students will perform MTM in the future.