Cloning, Sequencing, and Characterization of Luciola italica Luciferase

The characteristic yellow-green light of a firefly is the result of a multi-step reaction catalyzed by the luciferase enzyme. This enzyme has many applications in the biomedical field and ongoing work is being done to alter its properties to better fit these applications. The purpose of this project was to clone the Luciola italica luciferase cDNA and to express, purify and fully characterize the corresponding bioluminescence-catalyzing enzyme in hopes of obtaining novel bioluminescent materials. Fireflies were collected in the countryside of Bologna, Italy, flash frozen in liquid nitrogen and total RNA was extracted from the firefly lanterns. The L. italica luciferase cDNA was successfully cloned by RT-PCR using a gene-specific primer set based on the DNA sequence of the Eastern European Luciola mingrelica luciferase gene. The L. italica cDNA was determined to be 1647 base pairs in length with an open reading frame of 548 amino acids. Initial characterization of the enzyme showed that the L. italica protein exhibits bioluminescent activity similar in intensity to the common North American Photinus pyralis luciferase; however it produces light that is slightly red-shifted (having maximum emission at 564 nm). By steady state kinetics analysis, the L. italica Km for LH2 was found to be 0.095 mM, and that of P. pyralis is 0.015 mM. On the converse, both enzymes had similar Km values for Mg-ATP (0.160 mM for P. pyralis and 0.180 mM for L. italica). The L. italica enzyme was found to sustain its light in the visible region for a longer period of time than the P. pyralis enzyme. Phylogenetic analysis showed that the L. italica luciferase gene has 95.8% and 95.6% amino acid sequence identity to the Hotaria unmunsana (Korea) and Hotaria parvula (Japan) luciferase proteins, respectively. The processes that were used to clone the Luciola italica luciferase gene, characterize the protein, and optimize protein growth conditions ar

A pharmacist-driven academic detailing program to increase adult pneumococcal vaccination

Objectives To describe our statewide, pharmacist-led education campaign to increase knowledge and awareness of pneumococcal immunization recommendations. Setting Immunization providers and residents in the state of Rhode Island. Practice description A clinical pathway (i.e., decision-support tool) was developed to educate health professionals about appropriate indications, administration schedules, and frequently asked questions for the 2 different adult pneumococcal vaccines. Academic detailing and distribution of the clinical pathway to health professionals was conducted across Rhode Island. Community outreach activities included radio ads as well as distribution of patient handouts and wallet cards at community events. Practice innovation To our knowledge, this was the first statewide, pharmacist-driven academic detailing and community outreach campaign to promote adult pneumococcal vaccination. Evaluation Academically detailed immunization providers received a 6-question survey. Pneumococcal disease rate differences between the study periods were evaluated with the use of Fisher exact tests, whereas changes in vaccination were assessed with the use of chi-square tests. Results From November 2013 through July 2015, our academic detailers visited and distributed our vaccination pathway materials to more than 400 practice sites across Rhode Island, including 68% of community pharmacies and all adult acute care hospitals. Of the 413 surveys completed, 92% of respondents agreed that their knowledge of the pneumococcal conjugate vaccine, 13-valent and pneumococcal polysaccharide vaccine, 23-valent had improved. Pneumococcal vaccination increased significantly (absolute difference 3.9%, percentage change in proportion 5.4%; P = 0.01), and pneumococcal disease decreased significantly between the preintervention and intervention periods (−2.74/10,000 discharges [95% CI −5.15 to −0.32], P = 0.02). Invasive pneumococcal disease decreased by 21 cases per 1,000,000 population per year between the preintervention and postintervention periods (−42.25 to 0.14, P = 0.05). Conclusion Our statewide, pharmacist-driven pneumococcal vaccination educational outreach program resulted in favorable provider feedback relative to knowledge change and perceptions. Vaccination increased and pneumococcal disease decreased during the study period

1437. Family Duty and Safety Linked to Overcoming Attitudinal Barriers to Adult Pneumococcal Vaccination in Disparate Populations

Background: Minority adult populations are at a higher risk for invasive pneumococcal disease and also have significantly lower vaccination rates when compared with the general population. Ingrained attitudes are a significant barrier to receipt of pneumococcal vaccine in these disparate populations, and therefore we tested targeted informational messaging to overcome these. Methods: A survey instrument of attitudinal questions related to pneumococcal vaccination was administered via YouGov, an online public national survey house in 2017. Socioeconomic information was captured and linked to baseline Likert scale attitudinal question responses. Respondents were randomly assigned into subsamples that received different science-based messages that included information on pneumococcal vaccines related to: pneumonia prevention, fatality/consequences, vaccine safety information, family duty/safety, and a combined vignette including all of these. Because of the random assignment, any differences observed in the respondents’ outcomes across subsamples can be attributed to the messages. Descriptive statistics were used to compare the persuasive effectiveness of these messages to conventional vaccine information across racial and ethnic groups. Results: A total of 2,608 respondents, 1,327 (51%) white and 1,281 (49%) non-white (over-sampled) were represented. Of the total respondents as well as in white, and non-white respondents, the combined vignette was associated with positive coefficients of b = 0.26, b = 0.24, and b = 0.32, respectively (P-values all \u3c0.05). In whites, the vaccine safety information and family duty/safety also had significant coefficients b = 0.24 ( = P = 0.012) and b = 0.24 (P = 0016), respectively. In non-Whites, family duty/safety was the only additional message with a significant coefficient b = 0.25 (P = 0.007). Conclusion: In this survey assessing attitudes toward pneumococcal vaccination across racial and ethnic subpopulations, the disparate population was persuaded to receive the vaccine only when family duty and safety were linked within the informational messages. Future studies implementing this informational messaging strategy should be performed to validate this finding

Oakland Cemetery Comfort Station Buildings

This Historic Structure Report attempts to define the historical context and physical condition of the women\u27s and men\u27s comfort stations at Oakland Cemetery. The comfort stations were constructed in 1908, fifty-eight years after the opening of Oakland, in order to provide adequate public restroom facilities for the large crowds who visited the cemetery during its early history. A group effort has been made to research and document the history of the two comfort buildings, assess their current status, and make recommendations for treatment.https://scholarworks.gsu.edu/history_heritagepreservation/1053/thumbnail.jp

Barriers to exercise in people with Parkinson disease

BACKGROUND: Exercise is known to reduce disability and improve quality of life in people with Parkinson disease (PD). Although barriers to exercise have been studied in older adults, barriers in people with chronic progressive neurological diseases, such as PD, are not well defined. OBJECTIVE: The purpose of this study was to identify perceived barriers to exercise in people with PD. DESIGN: The study had a cross-sectional design. METHODS: People who had PD, dwelled in the community, and were at stage 2.4 on the Hoehn and Yahr scale participated in this cross-sectional study (N=260; mean age=67.7 years). Participants were divided into an exercise group (n=164) and a nonexercise group (n=96). Participants self-administered the barriers subscale of the Physical Fitness and Exercise Activity Levels of Older Adults Scale, endorsing or denying specific barriers to exercise participation. Multivariate logistic regression analysis was used to examine the contribution of each barrier to exercise behavior, and odds ratios were reported. RESULTS: Three barriers were retained in the multivariate regression model. The nonexercise group had significantly greater odds of endorsing low outcome expectation (ie, the participants did not expect to derive benefit from exercise) (odds ratio [OR]=3.93, 95% confidence interval [CI]=2.08–7.42), lack of time (OR=3.36, 95% CI=1.55–7.29), and fear of falling (OR=2.35, 95% CI=1.17–4.71) than the exercise group. LIMITATIONS: The cross-sectional nature of this study limited the ability to make causal inferences. CONCLUSIONS: Low outcome expectation from exercise, lack of time to exercise, and fear of falling appear to be important perceived barriers to engaging in exercise in people who have PD, are ambulatory, and dwell in the community. These may be important issues for physical therapists to target in people who have PD and do not exercise regularly. The efficacy of intervention strategies to facilitate exercise adherence in people with PD requires further investigation

The rehabilitation enhancing aging through connected health (REACH) study: study protocol for a quasi-experimental clinical trial

Background: Mobility limitations among older adults increase the risk for disability and healthcare utilization. Rehabilitative care is identified as the most efficacious treatment for maintaining physical function. However, there is insufficient evidence identifying a healthcare model that targets prevention of mobility decline among older adults. The objective of this study is to evaluate the preliminary effectiveness of a physical therapy program, augmented with mobile tele-health technology, on mobility function and healthcare utilization among older adults. Methods: This is a quasi-experimental 12-month clinical trial conducted within a metropolitan-based healthcare system in the northeastern United States. It is in parallel with an existing longitudinal cohort study evaluating mobility decline among community-dwelling older adult primary care patients over one year. Seventy-five older adults (≥ 65–95 years) are being recruited using identical inclusion/exclusion criteria to the cohort study. Three aims will be evaluated: the effect of our program on 1) physical function, 2) healthcare utilization, and 3) healthcare costs. Changes in patient-reported function over 1 year in those receiving the intervention (aim 1) will be compared to propensity score matched controls (N = 150) from the cohort study. For aims 2 and 3, propensity scores, derived from logistic regression model that includes demographic and diagnostic information available through claims and enrollment information, will be used to match treatment and control patients in a ratio of 1:2 or 1:3 from a Medicare Claims Registry derived from the same geographic region. The intervention consists of a one-year physical therapy program that is divided between a combination of outpatient and home visits (6–10 total visits) and is augmented on a computerized tablet using of a commercially available application to deliver a progressive home-based exercise program emphasizing lower-extremity function and a walking program. Discussion Incorporating mobile health into current healthcare models of rehabilitative care has the potential to decrease hospital visits and provide a longer duration of care. If the hypotheses are supported and demonstrate improved mobility and reduced healthcare utilization, this innovative care model would be applicable for optimizing the maintenance of functional independence among community-dwelling older adults. Trial registration ClinicalTrial.gov Identifier: NCT02580409 (Date of registration October 14, 2015)

Genomic profiling distinguishes familial multiple and sporadic multiple meningiomas

<p>Abstract</p> <p>Background</p> <p>Meningiomas may occur either as familial tumors in two distinct disorders, familial multiple meningioma and neurofibromatosis 2 (NF2), or sporadically, as either single or multiple tumors in individuals with no family history. Meningiomas in NF2 and approximately 60% of sporadic meningiomas involve inactivation of the <it>NF2 </it>locus, encoding the tumor suppressor merlin on chromosome 22q. This study was undertaken to establish whether genomic profiling could distinguish familial multiple meningiomas from sporadic solitary and sporadic multiple meningiomas.</p> <p>Methods</p> <p>We compared 73 meningiomas presenting as sporadic solitary (64), sporadic multiple (5) and familial multiple (4) tumors using genomic profiling by array comparative genomic hybridization (array CGH).</p> <p>Results</p> <p>Sporadic solitary meningiomas revealed genomic rearrangements consistent with at least two mechanisms of tumor initiation, as unsupervised cluster analysis readily distinguished tumors with chromosome 22 deletion (associated with loss of the <it>NF2 </it>tumor suppressor) from those without chromosome 22 deletion. Whereas sporadic meningiomas without chromosome 22 loss exhibited fewer chromosomal imbalance events overall, tumors with chromosome 22 deletion further clustered into two major groups that largely, though not perfectly, matched with their benign (WHO Grade I) or advanced (WHO Grades II and III) histological grade, with the latter exhibiting a significantly greater degree of genomic imbalance (P < 0.001). Sporadic multiple meningiomas showed a frequency of genomic imbalance events comparable to the atypical grade solitary tumors. By contrast, familial multiple meningiomas displayed no imbalances, supporting a distinct mechanism for the origin for these tumors.</p> <p>Conclusion</p> <p>Genomic profiling can provide an unbiased adjunct to traditional meningioma classification and provides a basis for exploring the different genetic underpinnings of tumor initiation and progression. Most importantly, the striking difference observed between sporadic and familial multiple meningiomas indicates that genomic profiling can provide valuable information for differential diagnosis of subjects with multiple meningiomas and for considering the risk for tumor occurrence in their family members.</p

Identification of a Rare Coding Variant in Complement 3 Associated with Age-related Macular Degeneration

Macular degeneration is a common cause of blindness in the elderly. To identify rare coding variants associated with a large increase in risk of age-related macular degeneration (AMD), we sequenced 2,335 cases and 789 controls in 10 candidate loci (57 genes). To increase power, we augmented our control set with ancestry-matched exome sequenced controls. An analysis of coding variation in 2,268 AMD cases and 2,268 ancestry matched controls revealed two large-effect rare variants; previously described R1210C in the CFH gene (fcase = 0.51%, fcontrol = 0.02%, OR = 23.11), and newly identified K155Q in the C3 gene (fcase = 1.06%, fcontrol = 0.39%, OR = 2.68). The variants suggest decreased inhibition of C3 by Factor H, resulting in increased activation of the alternative complement pathway, as a key component of disease biology

Preterm neonatal morbidity and mortality by gestational age: a contemporary cohort

Although preterm birth less than 37 weeks gestation is the leading cause of neonatal morbidity and mortality in the United States, the majority of data regarding preterm neonatal outcomes come from older studies, and many reports have been limited to only very preterm neonates. Delineation of neonatal outcomes by delivery gestational age is needed to further clarify the continuum of mortality and morbidity frequencies among preterm neonates