Comparison and Analysis of ADH Isozymes from a Population of Chrysoma pauciflosculosa

Chrysoma pauciflosculosa is a rare evergreen shrub that is located on a sandy ridge. There are two populations of Chrysoma located in Pembroke, North Carolina. No published information on isozyme studies or population genetics exists for this species. Isozymes are variable forms of an enzyme. Two enzymes, ADH and MDH, were chosen for the comparison. Two prior students at UNCP developed an initial protocol for studying isozymes in this species. There data was used for storage and grinding. The leaves were collected from one site near Purnell Swett High School in Pembroke, North Carolina. The leaves were homogenized, electrophoresed on acrylamide gels and stained for the particular enzyme being studied. It was determined that ADH was active in this population of Chrysoma

Strategy application, observability, and the choice combinator.

In many strategic systems, the choice combinator provides a powerful mechanism for controlling the application of rules and strategies to terms. The ability of the choice combinator to exercise control over rewriting is based on the premise that the success and failure of strategy application can be observed. In this paper we present a higher-order strategic framework with the ability to dynamically construct strategies containing the choice combinator. To this framework, a combinator called hide is introduced that prevents the successful application of a strategy from being observed by the choice combinator. We then explore the impact of this new combinator on a real-world problem involving a restricted implementation of the Java Virtual Machine

New leads against drug-resistant organisms : shifting the focus in discovery towards rapid phenotypic screening and mechanism of action

Over the past century, antibiotic therapy has been successful in nearly eliminating a former leading cause of death. Mortality records from the past 100 years in the US show a decrease from 46% of deaths due to infectious diseases, to just 3% in 2010. While the use of antibiotics has had a tremendously positive impact on healthcare, the subsequent rise of antibiotic resistance in pathogens has threatened to reverse recent advances in public health and the study of antibiotic mechanisms has gained significant interest. Issues leading to emergence of antibiotic resistant pathogens include overprescribing of antibiotics and abuses by the agricultural industry, with 50% of manufactured antibiotics used in agriculture, rather than the medical field as originally intended. Misuse, by failure to regard antibiotics as an inestimable resource, coupled with stagnation in discovery of new antibiotics, has led to the prevalence of antibiotic-resistant pathogens. Approaches to combat drug resistant pathogens include large scale screening of natural sources, such as plants and fungi, for antimicrobial compounds with diverse chemical scaffolds and prioritization of antimicrobials operating by mechanisms of action which bacteria encounter infrequently. Due to the emergence of resistance, the World Health Organization considers Gram-negative pathogen Acinetobacter baumannii a top priority for therapeutic development. Using this priority pathogen and a phenotypic, agar plate-based assay, a unique library of extracts from 2,500 diverse fungi was screened for antimicrobial activity against a highly virulent, drug resistant strain of A. baumannii (AB5075). The most potent hit from this screen was an extract from the fungi Tolypocladium sp., which was found to produce pyridoxatin. Another active extract from the fungi Trichoderma deliquescens was characterized and yielded Trichokonin VII and Trichokonin VIII. Evaluation of pyridoxatin against A. baumannii (AB5075) in a broth microdilution assay revealed a minimum inhibitory concentration (MIC) of 38.0µM, which was the strongest lead compared to the known antibiotic levofloxacin MIC of 27.7µM. Mass spectrometry, Marfey’s analysis and nuclear magnetic resonance spectroscopy were utilized to confirm the structures of Trichokonin VII and Trichokonin VIII in comparison to previous reports. In an in vivo Galleria mellonella model, pyridoxatin tested at 150 mg/kg exhibited minimal toxicity (90% survival) and promising antimicrobial efficacy (50% survival) after 5 days. Trichokonin VII and Trichokonin VIII tested at 150 mg/kg were toxic, with 20% survival and 40% survival after 5 days, respectively. The findings of this project suggest that pyridoxatin may serve as a lead compound for the development of antimicrobials against A. baumannii. They also demonstrate the value of the phenotypic screening approach employed here to uncover diverse lead antimicrobial compounds. A second approach to counteracting antimicrobial resistance was taken with a botanical extract mechanism of action classification prediction method that was developed using metabolomics in the model antimicrobial system of Hypericum calycinum (creeping St. John’s wort) against Methicillin-resistant Staphylococcus aureus (MRSA). Antimicrobial botanicals, such as H. calycinum, exhibit unique mechanisms of action due to a complex array of compounds and combination effects which challenges bacteria in developing resistance. MRSA strain USA300 LAC AH1263, a model organism we observed with susceptibility to H. calycinum, is a clinically relevant drug-resistant pathogen rapidly developing resistance. To evaluate these qualities, metabolomics analyses were conducted on the spent media from MRSA cultures incubated with various treatments. The treatments included sub-minimum inhibitory concentrations (sub-MIC) of six clinically relevant antibiotics representing three mechanisms of action. Also included in the study were a H. calycinum extract, an H. calycinum active fraction, or the pure botanical compound hyperforin commonly found in Hypericum calycinum. Previously developed metabolomics data processing techniques were refined for evaluating profiles of the spent media and narrowing down features detected across all samples from 8,900 features to a list of 32 unique features (ions detected by the mass spectrometer) associated with mechanism of action. Further investigation of the identities of these features and biochemical studies to confirm the antimicrobial mechanism of action of H. calycinum against MRSA are necessary to validate the model in predicting antimicrobial mechanism of action in botanical extracts

Dependable software through higher-order strategic programming.

Program transformation is a restricted form of software construction that can be amenable to formal verification. When successful, the nature of the evidence provided by such a verification is considered strong and can constitute a major component of an argument that a high-consequence or safety-critical system meets its dependability requirements. This article explores the application of novel higher-order strategic programming techniques to the development of a portion of a class loader for a restricted implementation of the Java Virtual Machine (JVM). The implementation is called the SSP and is intended for use in high-consequence safety-critical embedded systems. Verification of the strategic program using ACL2 is also discussed

Higher-order transformation and the distributed data problem.

The distributed data problem, is characterized by the desire to bring together semantically related data from syntactically unrelated portions of a term. Two strategic combinators, dynamic and transient, are introduced in the context of a classical strategic programming framework. The impact of the resulting system on instances of the distributed data problem is then explored

A parasite-derived 68-mer peptide ameliorates autoimmune disease in murine models of Type 1 diabetes and multiple sclerosis

© 2016 cThe Author(s). Helminth parasites secrete molecules that potently modulate the immune responses of their hosts and, therefore, have potential for the treatment of immune-mediated human diseases. FhHDM-1, a 68-mer peptide secreted by the helminth parasite Fasciola hepatica, ameliorated disease in two different murine models of autoimmunity, type 1 diabetes and relapsing-remitting immune-mediated demyelination. Unexpectedly, FhHDM-1 treatment did not affect the proliferation of auto-antigen specific T cells or their production of cytokines. However, in both conditions, the reduction in clinical symptoms was associated with the absence of immune cell infiltrates in the target organ (islets and the brain tissue). Furthermore, after parenteral administration, the FhHDM-1 peptide interacted with macrophages and reduced their capacity to secrete pro-inflammatory cytokines, such as TNF and IL-6. We propose this inhibition of innate pro-inflammatory immune responses, which are central to the initiation of autoimmunity in both diseases, prevented the trafficking of autoreactive lymphocytes from the periphery to the site of autoimmunity (as opposed to directly modulating their function per se), and thus prevented tissue destruction. The ability of FhHDM-1 to modulate macrophage function, combined with its efficacy in disease prevention in multiple models, suggests that FhHDM-1 has considerable potential as a treatment for autoimmune diseases

Lack of trust in maternal support is associated with negative interpretations of ambiguous maternal behavior

Attachment theory assumes that children who lack trust in maternal availability for support are more inclined to interpret maternal behavior in congruence with their expectation that mother will remain unavailable for support. To provide the first test of this assumption, early adolescents (9-13 years old) were asked to assess whether ambiguous interactions with mother should be interpreted in a positive or a negative way. In our sample (n = 322), results showed that early adolescents' lack of trust in their mother's availability for support was related to more negative interpretations of maternal behavior. The associations remained significant after controlling for depressive mood. The importance of these findings for our understanding of attachment theory, attachment stability, and clinical practice are discussed

Extracellular Hsp72 concentration relates to a minimum endogenous criteria during acute exercise-heat exposure

Extracellular heat-shock protein 72 (eHsp72) concentration increases during exercise-heat stress when conditions elicit physiological strain. Differences in severity of environmental and exercise stimuli have elicited varied response to stress. The present study aimed to quantify the extent of increased eHsp72 with increased exogenous heat stress, and determine related endogenous markers of strain in an exercise-heat model. Ten males cycled for 90 min at 50% O2peak in three conditions (TEMP, 20°C/63% RH; HOT, 30.2°C/51%RH; VHOT, 40.0°C/37%RH). Plasma was analysed for eHsp72 pre, immediately post and 24-h post each trial utilising a commercially available ELISA. Increased eHsp72 concentration was observed post VHOT trial (+172.4%) (P<0.05), but not TEMP (-1.9%) or HOT (+25.7%) conditions. eHsp72 returned to baseline values within 24hrs in all conditions. Changes were observed in rectal temperature (Trec), rate of Trec increase, area under the curve for Trec of 38.5°C and 39.0°C, duration Trec ≥ 38.5°C and ≥ 39.0°C, and change in muscle temperature, between VHOT, and TEMP and HOT, but not between TEMP and HOT. Each condition also elicited significantly increasing physiological strain, described by sweat rate, heart rate, physiological strain index, rating of perceived exertion and thermal sensation. Stepwise multiple regression reported rate of Trec increase and change in Trec to be predictors of increased eHsp72 concentration. Data suggests eHsp72 concentration increases once systemic temperature and sympathetic activity exceeds a minimum endogenous criteria elicited during VHOT conditions and is likely to be modulated by large, rapid changes in core temperature

Flower Bats (Glossophaga soricina) and Fruit Bats (Carollia perspicillata) Rely on Spatial Cues over Shapes and Scents When Relocating Food

Natural selection can shape specific cognitive abilities and the extent to which a given species relies on various cues when learning associations between stimuli and rewards. Because the flower bat Glossophaga soricina feeds primarily on nectar, and the locations of nectar-producing flowers remain constant, G. soricina might be predisposed to learn to associate food with locations. Indeed, G. soricina has been observed to rely far more heavily on spatial cues than on shape cues when relocating food, and to learn poorly when shape alone provides a reliable cue to the presence of food.Here we determined whether G. soricina would learn to use scent cues as indicators of the presence of food when such cues were also available. Nectar-producing plants fed upon by G. soricina often produce distinct, intense odors. We therefore expected G. soricina to relocate food sources using scent cues, particularly the flower-produced compound, dimethyl disulfide, which is attractive even to G. soricina with no previous experience of it. We also compared the learning of associations between cues and food sources by G. soricina with that of a related fruit-eating bat, Carollia perspicillata. We found that (1) G. soricina did not learn to associate scent cues, including dimethyl disulfide, with feeding sites when the previously rewarded spatial cues were also available, and (2) both the fruit-eating C. perspicillata and the flower-feeding G. soricina were significantly more reliant on spatial cues than associated sensory cues for relocating food.These findings, taken together with past results, provide evidence of a powerful, experience-independent predilection of both species to rely on spatial cues when attempting to relocate food