The relationship between alkaloid-based chemical defenses and diet in dendrobatid poison frogs

Chemical defenses are common among organisms and represent some of the most complex adaptations for avoiding predation, yet our understanding of the ecological nature of these systems remains incomplete. Poison frogs are a group of chemically defended organisms that are dependent entirely on diet for chemical defense. In this study, I identified the dietary arthropods responsible for chemical defense in poison frogs, described spatial and temporal patterns in alkaloid composition of poison frogs, and established links between patterns of variation in alkaloid defense and arthropod diet in poison frogs. Identifying dietary sources and studying patterns of variation in alkaloid composition is fundamental to understanding the ecology and evolution of chemical defense in poison frogs. ^ The dendrobatid poison frog Oophaga pumilio shares many alkaloids in common with other poison frogs and is known to vary in alkaloid composition throughout its geographic range. I designed my dissertation to take advantage of these characteristics and use O. pumilio as a model species for the study of chemical defense in poison frogs. Here, I identified siphonotid millipedes as a source for spiropyrrolizidine alkaloids, formicine ants as a source for pumiliotoxin alkaloids, and oribatid mites as dietary sources for the majority of alkaloids found in poison frogs. I found that alkaloid composition varied spatially and temporally, on both small and large scales, within and among populations of O. pumilio. Alkaloid variation between populations was related to geographic distance, and closer populations tended to have alkaloid compositions more similar to each other than to distant populations. ^ The findings of my study suggest that oribatid mites are the most important dietary source of alkaloids in poison frogs. However, overall alkaloid defense in poison frogs is based on a combination of dietary arthropods, including mites, ants, millipedes, and beetles. Variation in chemical defenses of poison frogs is due to (1) spatial and temporal differences in the presence of alkaloids in certain arthropods and (2) differences in the availability of certain alkaloid-containing arthropods, which are likely the result of differences as well as successional changes in forest structure among locations and through time. ^

Identification of Anziaic Acid, a Lichen Depside from Hypotrachyna sp., as a New Topoisomerase Poison Inhibitor

Topoisomerase inhibitors are effective for antibacterial and anticancer therapy because they can lead to the accumulation of the intermediate DNA cleavage complex formed by the topoisomerase enzymes, which trigger cell death. Here we report the application of a novel enzyme-based high-throughput screening assay to identify natural product extracts that can lead to increased accumulation of the DNA cleavage complex formed by recombinant Yersinia pestistopoisomerase I as part of a larger effort to identify new antibacterial compounds. Further characterization and fractionation of the screening positives from the primary assay led to the discovery of a depside, anziaic acid, from the lichen Hypotrachyna sp. as an inhibitor for both Y. pestis and Escherichia colitopoisomerase I. In in vitro assays, anziaic acid exhibits antibacterial activity against Bacillus subtilis and a membrane permeable strain of E. coli. Anziaic acid was also found to act as an inhibitor of human topoisomerase II but had little effect on human topoisomerase I. This is the first report of a depside with activity as a topoisomerase poison inhibitor and demonstrates the potential of this class of natural products as a source for new antibacterial and anticancer compounds.

Ontogenetic Scaling of Poison Glands in Dendrobates Pumilio

The relationship between granular (poison) gland size and density was examined in an ontogenetic series of the strawberry dart-poison frog, Dendrobates pumilio. Specimens used in this study were collected from the La Selva Biological Station in northeastern Costa Rica. Patches of skin from the dorsal surface of seven frogs, ranging in size from 11 to 23 mm snout-vent length (SVL), were fixed and embedded in paraffin for histological sectioning. Poison gland size and density were quantified microscopically in these sections. Poison glands are uniformly distributed across the skin and mean poison gland diameter increases at a rate faster than snout-vent length from 42.5 at SVL 11mm to 120.0 at SVL 23 mm. Conversely, gland density decreases with body size from 71.9 glands/mm2 to 33.2 glands/mm2 • Due to the positive allometric growth of the poison glands, the percentage of skin surface occupied by poison glands increases from 10.1-22.1% in small frogs (SVL<18 >mm) to 50.0-65.2% in large frogs (SVL>19MM), resulting in more toxin per mm2 in the larger animals. The largest increase in toxicity is correlated temporally with the onset of sexual maturity rather than with changes in aposematic coloring.

Mechanisms of chloroperoxidase-catalyzed enantioselective reactions as probed by site-directed mutagenesis and isotopic labeling

Chloroperoxidase (CPO) is a heme-containing glycoprotein secreted by the marine fungus Caldariomyces fumago. Chloroperoxidase contains one ferriprotoporphyrin IX prosthetic group per molecule and catalyzes a variety of reactions, such as halogenation, peroxidation and epoxidation. The versatile catalytic activities of CPO coupled with the increasing demands for chiral synthesis have attracted an escalating interest in understanding the mechanistic and structural properties of this enzyme. In order to better understand the mechanisms of CPO-catalyzed enantioselective reactions and to fine-tune the catalytic properties of chloroperoxidase, asparagine 74 (N74) located in the narrow substrate access channel of CPO was replaced by a bulky, nonpolar valine and a polar glutamine using site-directed mutagenesis. The CPO N74 mutants displayed significantly enhanced activity toward nonpolar substrates compared to wild-type CPO as a result of changes in space and polarity of the heme distal environment. More interestingly, N74 mutants showed dramatically decreased chlorination and catalase activity but significantly enhanced epoxidation activity as a consequence of improved kinetic perfection introduced by the mutation as reflected by the favorable changes in k cat and kcat/KM of these reactions. It is also noted that the N74V mutant is capable of decomposing cyanide, the most notorious poison for many hemoproteins, as judged by the unique binding behavior of N74V with potassium cyanide. Histidine 105 (H105) was replaced by a nonpolar amino acid alanine using site-directed mutagenesis. The CPO H105 mutant (H105A) displayed dramatically decreased chlorination and catalase activity possibly because of the decreased polarity in the heme distal environment and loss of the hydrogen bonds between histidine 105 and glutamic acid 183. However, significantly increased enantioselectivity was observed for the epoxidation of bulky styrene derivatives. Furthermore, my study provides strong evidence for the proposed histidine/cysteine ligand switch in chloroperoxidase, providing experimental support for the structure of the 420-nm absorption maximum for a number of carbon monoxide complexes of heme-thiolate proteins. For the NMR study, [dCPO(heme)] was produced using 90% deuterated growth medium with excess heme precursors and [dCPO(Phe)] was grown in the same highly deuterated medium that had been supplemented with excess natural phenylalanine. To make complete heme proton assignments, NMR spectroscopy has been performed for high-resolution structural characterization of [dCPO(heme)] and [dCPO(Phe)] to achieve unambiguous and complete heme proton assignments, which also allows important amino acids close to the heme active center to be determined.

Beginning to Explore Dashboard Dining

It can be nutritious and healthy if done right. Fruits and vegetables, a granola bar, smoothie, or some fresh squeezed Florida orange juice would be good choices. On the other hand, it can poison you. Perishable protein and dairy products must be packed in a well- insulated cooler with plenty of ice and a refrigerator thermometer kept inside to en-sure the food stays below 40 degrees Fahrenheit. If you are not completely safe, it can kill you. According to Hagerty Insurance of Traverse City, Michigan, the top ten worst foods to consume are coffee, hot soups, tacos, chili, juicy hamburgers, fried chicken, any barbecued food, filled doughnuts, soft drinks, and chocolate. (see Lisa Chin, 2003) It simply takes a sudden scalding spill, an unexpected splash, or dripping condiments, any of which demand your immediate attention, to become an instant fatality.

Restructuring In The Hospitality Industry

In her dialogue entitled - Restructuring in the Hospitality Industry - Elisa S. Moncarz, Associate Professor, the School of Hospitality Management at Florida International University, intends for you to know the following: “Recent years have seen a proliferation of restructurings of major American corporations creating an extremely important issue that has affected U.S. business. This article discusses restructuring issues in the hospitality industry, focusing attention on its causes and motivations, as well as on its benefits and perils. The author considers the impact of restructuring on investors and management while examining recent restructurings involving hospitality firms.” In defining the concept of restructuring, Associate Professor Moncarz informs you, “Restructuring entails the implementation of fundamental and comprehensive modification of a company's operational and/or financial structure.” “It has, indeed, become fashionable to take a company apart and put it back together in a different form,” the author says. Additionally, Moncarz refers to a Wall Street Journal study, dated August 1985, which reveals that nearly half the large American corporations were, or were soon to be restructured in the 1984/85 time frame. There are several distinct types of restructurings and the author wants you to be aware of some of them. “…threats of takeover attempts, the larger part of all restructuring have been initiated willingly in order to expand or divest a company's line of business (i.e., operational restructurings) or redirect its finances (i.e., financial restructurings),” the author reveals. “Two principal types of operational restructurings are mergers and acquisitions [M&A], and divestitures [disposing of unwanted units or assets],” Moncarz further defines the concepts of expansion and divestiture. The author explains several types of financial restructuring sketches used in the hospitality industry, including stock re-purchasing, debt issuances and redemptions, swapping debt for equity, and effective theories of realigning debt through extending loans and/or revising terms. To expand their businesses, Moncarz makes anecdotal reference to several major food and beverage corporations that have successfully employed operational restructuring principles. The author wades into the shallow end of the hostile takeover pool by explaining some of the corporate restructuring concepts used to repel that aggressive technique. Walt Disney Company completely redesigned their entire upper level management structure in a successful effort to thwart a hostile takeover bid by corporate raider Saul P. Steinberg, Moncarz informs. To close, the author touches on leveraged buyouts [LBOs], and stock repurchases to divest unwanted divisions and immobilize hostile takeover attempts. A lengthy table of - Selected Restructurings in the Hospitality Industry [1982 to date of article] – is also included.

Stray dogs in Bucharest, and the political economy of Romanian socialism. From hunting the “enemies” of Stalinist economy, to poisoning urban parasites, and the rise of urban anarchy: 1950s-1990s.

My paper discusses three different ways in which stray dogs have been intertwined with ideologies of economic and urban development in Romania. I categorize results from archival and ethnographic research under three major time periods: early socialism, late socialism, and post-socialism. During early socialism stray dogs were seen to be damaging the soviet economy by killing species that humans could also hunt, like rabbits. During late socialism, stray dogs appeared as the enemies of the communist city, and the department of urban sanitation was given orders to poison dogs with strychnine. Finally, the increasing number of stray dogs in Bucharest after the collapse of communism was seen as a direct result of former communist demolitions, and was also taken as a sign of the collapsing state. Through such examples my paper discusses how the state and particular population groups have seen dogs as parts of an unwanted and dangerous nature, rather than a species that needs to be protected. I argue that distinctions of nature and culture have served discourses of civilization and the view of Bucharest as a model socialist, and then European city. Throughout my paper I juxtapose the treatment of stray dogs with other, more “valued” urban natures like the protection of parks, the wide-spread hobby of pigeon breeding during socialist years, the most recent debate on saving the rural area of Rosia Montana from non-environmentally friendly methods of gold extraction, and the current trend of healthy eating and living.

Mechanisms of Chloroperoxidase-catalyzed Enantioselective Reactions as Probed by Site-directed Mutagenesis and Isotopic Labeling

Chloroperoxidase (CPO) is a heme-containing glycoprotein secreted by the marine fungus Caldariomyces fumago. Chloroperoxidase contains one ferriprotoporphyrin IX prosthetic group per molecule and catalyzes a variety of reactions, such as halogenation, peroxidation and epoxidation. The versatile catalytic activities of CPO coupled with the increasing demands for chiral synthesis have attracted an escalating interest in understanding the mechanistic and structural properties of this enzyme. In order to better understand the mechanisms of CPO-catalyzed enantioselective reactions and to fine-tune the catalytic properties of chloroperoxidase, asparagine 74 (N74) located in the narrow substrate access channel of CPO was replaced by a bulky, nonpolar valine and a polar glutamine using site-directed mutagenesis. The CPO N74 mutants displayed significantly enhanced activity toward nonpolar substrates compared to wild-type CPO as a result of changes in space and polarity of the heme distal environment. More interestingly, N74 mutants showed dramatically decreased chlorination and catalase activity but significantly enhanced epoxidation activity as a consequence of improved kinetic perfection introduced by the mutation as reflected by the favorable changes in kcat and kcat/KM of these reactions. It is also noted that the N74V mutant is capable of decomposing cyanide, the most notorious poison for many hemoproteins, as judged by the unique binding behavior of N74V with potassium cyanide. Histidine 105 (H105) was replaced by a nonpolar amino acid alanine using site-directed mutagenesis. The CPO H105 mutant (H105A) displayed dramatically decreased chlorination and catalase activity possibly because of the decreased polarity in the heme distal environment and loss of the hydrogen bonds between histidine 105 and glutamic acid 183. However, significantly increased enantioselectivity was observed for the epoxidation of bulky styrene derivatives. Furthermore, my study provides strong evidence for the proposed histidine/cysteine ligand switch in chloroperoxidase, providing experimental support for the structure of the 420-nm absorption maximum for a number of carbon monoxide complexes of heme-thiolate proteins. For the NMR study, [dCPO(heme)] was produced using 90% deuterated growth medium with excess heme precursors and [dCPO(Phe)] was grown in the same highly deuterated medium that had been supplemented with excess natural phenylalanine. To make complete heme proton assignments, NMR spectroscopy has been performed for high-resolution structural characterization of [dCPO(heme)] and [dCPO(Phe)] to achieve unambiguous and complete heme proton assignments, which also allows important amino acids close to the heme active center to be determined.