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Lipids are an important component of meat, as they provide desirable sensory characteristics and nutritional benefits. However, lipids are susceptible to degradation through oxidation and produce toxic oxidative byproducts. 4-Oxo-2-nonenal (4-ONE) is an oxidative byproduct that is highly reactive and cytotoxic. In this study, we investigated the influence of 4-ONE-induced protein degradation on fresh and gastric digested bovine skeletal muscle proteins. The results indicated that 4-ONE naturally forms in fresh muscle proteins. We report here for the first time that 4-ONE causes severe degradation of bovine skeletal muscle proteins. An SDS-PAGE gel analysis showed evidence that the skeletal muscle proteins attenuated over the incubation time, as the density of the protein bands faded significantly after 120 h. Additionally, protein and band density analyses showed a significant decrease in protein abundance and band densities throughout the incubation time. This study revealed that the lipid oxidation byproduct, 4-oxo-2-nonenal (4-ONE) is responsible for causing skeletal muscle protein degradation. Future studies should assess the bioprotective role of antioxidants and other food ingredients for their potential to prevent the formation and/or detoxification of 4-ONE in meat.

Multiple rice GT61 members were demonstrated to be xylan arabinosyltransferases (XATs) mediating 3-O-arabinosylation of xylan and the functions of XATs and xylan 2-O-xylosyltransferases were shown to be conserved in grass species. Xylan is the major hemicellulose in the cell walls of grass species and it is typified by having arabinofuranosyl (Araf) substitutions. In this report, we demonstrated that four previously uncharacterized, Golgi-localized glycosyltransferases residing in clade A or B of the rice GT61 family were able to mediate 3-O-arabinosylation of xylan when heterologously expressed in the Arabidopsis gux1/2/3 triple mutant. Biochemical characterization of their recombinant proteins established that they were xylan arabinosyltransferases (XATs) capable of transferring Araf residues onto xylohexaose acceptors, and thus they were named OsXAT4, OsXAT5, OsXAT6 and OsXAT7. OsXAT5 and the previously identified OsXAT2 were shown to be able to arabinosylate xylooligomers with a degree of polymerization of as low as 3. Furthermore, a number of XAT homologs from maize, sorghum, Brachypodium and switchgrass were found to exhibit activities catalyzing Araf transfer onto xylohexaose, indicating that they are XATs involved in xylan arabinosylation in these grass species. Moreover, we revealed that homologs of another GT61 member, xylan 2-O-xylosyltransferase (XYXT1), from these grass species could mediate 2-O-xylosylation of xylan when expressed in the Arabidopsis gux1/2/3 mutant. Together, our findings indicate that multiple OsXATs are involved in 3-O-arabinosylation of xylan and the functions of XATs and XYXTs are conserved in grass species.

Paclitaxel is a well-known cancer drug that functions as a mitotic inhibitor. This work focuses on a copper based crystal that encapsulates the pharmaceutical agent and serves as a drug delivery agent. A Copper10-Pacitaxil1 chloride (CU10PAC1) complex is synthesized and tested against the National Cancer Institute’s sixty cell line panel. The 10:1 ratio results in a crystal that was examined by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spec (MALDI-TOF-MS), Scanning Electron Microscopy (SEM) and Proton (1H) and Carbon (13C) Nuclear Magnetic Resonance (NMR). The potential attributes of a copper based crystal as an in vivo drug carrier for Paclitaxel are discussed.

Xyloglucan is the most abundant hemicellulose in the primary cell walls of dicots. Dicot xyloglucan is the XXXG type consisting of repeating units of three consecutive xylosylated Glc residues followed by one unsubstituted Glc. Its xylosylation is catalyzed by xyloglucan 6-xylosyltransferases (XXTs) and there exist five XXTs (AtXXT1-5) in Arabidopsis. While AtXXT1 and AtXXT2 have been shown to add the first two Xyl residues in the XXXG repeat, which XXTs are responsible for the addition of the third Xyl residue remains elusive although AtXXT5 was a proposed candidate. In this report, we generated recombinant proteins of all five Arabidopsis XXTs and one rice XXT (OsXXT1) in the mammalian HEK293 cells and investigated their ability to sequentially xylosylate Glc residues to generate the XXXG xylosylation pattern. We found that like AtXXT1/2, AtXXT4 and OsXXT1 could efficiently xylosylate the cellohexaose (G6) acceptor to produce mono- and di-xylosylated G6, whereas AtXXT5 was only barely capable of adding one Xyl onto G6. When AtXXT1-catalyzed products were used as acceptors, AtXXT1/2/4 and OsXXT1, but not AtXXT5, were able to xylosylate additional Glc residues to generate tri- and tetra-xylosylated G6. Further characterization of the tri- and tetra-xylosylated G6 revealed that they had the sequence of GXXXGG and GXXXXG with three and four consecutive xylosylated Glc residues, respectively. In addition, we have found that although tri-xylosylation occurred on G6, cello-oligomers with a degree of polymerization of 3 to 5 could only be mono- and di-xylosylated. Together, these results indicate that each of AtXXT1/2/4 and OsXXT1 is capable of sequentially adding Xyl onto three contiguous Glc residues to generate the XXXG xylosylation pattern and these findings provide new insight into the biochemical mechanism underlying xyloglucan biosynthesis.

Xyloglucan is a major hemicellulose in plant cell walls and exists in two distinct types, XXXG and XXGG. While the XXXG-type xyloglucan from dicot species only contains O-acetyl groups on side-chain galactose (Gal) residues, the XXGG-type xyloglucan from Poaceae (grasses) and Solanaceae bears O-acetyl groups on backbone glucosyl (Glc) residues. Although O-acetyltransferases responsible for xyloglucan Gal acetylation have been characterized, the biochemical mechanism underlying xyloglucan backbone acetylation remains to be elucidated. In this study, we showed that recombinant proteins of a group of DUF231 members from rice and tomato were capable of transferring acetyl groups onto O-6 of Glc residues in cello-oligomer acceptors, indicating that they are xyloglucan backbone 6-O-acetyltransferases (XyBATs). We further demonstrated that XyBAT-acetylated cellohexaose oligomers could be readily xylosylated by AtXXT1 (Arabidopsis xyloglucan xylosyltransferase 1) to generate acetylated, xylosylated cello-oligomers, whereas AtXXT1-xylosylated cellohexaose oligomers were much less effectively acetylated by XyBATs. Heterologous expression of a rice XyBAT in Arabidopsis led to a severe reduction in cell expansion and plant growth and a drastic alteration in xyloglucan xylosylation pattern with the formation of acetylated XXGG-type units, including XGG, XGGG, XXGG, XXGG,XXGGG and XXGGG (G denotes acetylated Glc). In addition, recombinant proteins of two Arabidopsis XyBAT homologs also exhibited O-acetyltransferase activity toward cellohexaose, suggesting their possible role in mediating xyloglucan backbone acetylation in vivo. Our findings provide new insights into the biochemical mechanism underlying xyloglucan backbone acetylation and indicate the importance of maintaining the regular xyloglucan xylosylation pattern in cell wall function.

The frontline tuberculosis (Tb) antibiotic isoniazid has been repurposed using a three component complex aimed at increasing the delivery efficiency and adding new avenues to its mechanism of action. This study focuses on pharmacokinetic studies of the isoniazid-sucrose–copper (II)-PEG-3350 complex. The assays include the Plasma Protein Binding Assay (85.8%), Caco-2 Permeability Assay (B→AP app , 0.13 × 10 −6 cm/s), Cytochrome P450 Inhibition Assay (i.e. CYP2B6, IC 50 = 7.26 μM), In vitro microsomal Stability Assay (t 1/2 NADPH-Dependent > 240 min), and HepG2 Cytotoxicity (no toxicity). The National Cancer Institute's 60 cell line panel is used to measure activity against cancer cells. The percent growth values averaged over all 60 cell lines indicates the complex has no anti-cancer activity, which also suggests a lack of general toxicity. It also provides data for the complexes specificity against Mycobacterium tuberculosis .

College athletic departments have a responsibility to provide a safe environment for student-athletes; however, most colleges do not have a crisis management plan that includes procedures for displaced student-athletes or alternate facilities to perform athletic events. Continuity of operations planning ensures athletic programs are equipped to maintain essential functions during, or shortly after, a disruption of operations due to possible hazards. Previous studies have identified a lack of emergency preparedness and continuity planning in college athletic departments. The purpose of this article is to illustrate in detail one approach to disaster planning for college athletic departments, namely the Federal Emergency Management Agency (FEMA) continuity of operations framework. By adhering to FEMA guidelines and promoting a best practices model, athletic programs can effectively plan to address potential hazards, as well as protect the organization's brand, image, and financial sustainability after a crisis event.

The nutritionally available pool of dissolved organic phosphorus (DOP) supports marine primary productivity in a range of ocean ecosystems but remains poorly resolved. Here, the relative lability of model phosphorous (P) compounds representing the major P(V) bond classes of marine DOP – phosphomonoesters (P-O-C) and phosphoanhydrides (P-O-P) – was assessed in diatom cultures of the genus Thalassiosira, as well as coastal field sites of the western North Atlantic. In diatom samples, maximum enzymatic hydrolysis rates revealed that the P-anhydride bonds of inorganic tripolyphosphate (3poly-P), followed by the P-anhydride bonds of adenosine 5’-triphosphate (ATP), were preferentially degraded relative to the P-monoesters adenosine 5’-monophosphate (AMP) and 4-methylumbelliferone phosphate (MUF-P). Consistent with these rate measurements, targeted proteomics analysis demonstrated that the underlying phosphatase diversity present in diatom samples was dominated by P-anhydride degrading enzymes (inorganic pyrophosphatases and nucleoside triphosphatases). Furthermore, biomass-normalized rates of ATP degradation were always suppressed under P-replete conditions in diatom cultures, but the effect of overall P availability on 3poly-P degradation was inconsistent among diatom strains, suggesting that inorganic polyphosphate (poly-P) degradation may persist irrespective of prevailing P levels in the marine environment. Indeed, the majority of field sites examined in the P-replete coastal western North Atlantic exhibited significantly higher maximum rates of inorganic poly-P hydrolysis relative to P-monoester hydrolysis, which was largely driven by phytoplankton dynamics. Based on these results, the possibility that P-anhydride utilization may contribute comparably or even more substantially than P-esters to community-level P demand, phytoplankton growth, and primary productivity should be considered.

Paclitaxel is a well-known cancer drug that functions as a mitotic inhibitor. This work focuses on a copper based crystal that encapsulates the pharmaceutical agent and serves as a drug delivery agent. A Copper

Methyl coenzyme M reductase (MCR) is a complex enzyme that catalyzes the final step in biological methanogenesis. To better understand its assembly, the recombinant MCR from the thermophile Methanothermococcus okinawensis (rMCR ok ) was expressed in the mesophile Methanococcus maripaludis . The rMCR ok was posttranslationally modified correctly and contained McrD and the unique nickel tetrapyrrole coenzyme F 430 . Subunits of the native M. maripaludis (MCR mar ) were largely absent, suggesting that the recombinant enzyme was formed by an assembly of cotranscribed subunits. Strong support for this hypothesis was obtained by expressing a chimeric operon comprising the His-tagged mcrA from M. maripaludis and the mcrBDCG from M. okinawensis in M. maripaludis . The His-tagged purified rMCR then contained the M. maripaludis McrA and the M. okinawensis McrBDG. The present study prompted us to form a working model for MCR assembly, which can be further tested by the heterologous expression system established here. IMPORTANCE Approximately 1.6% of the net primary production of plants, algae, and cyanobacteria are processed by biological methane production in anoxic environments. This accounts for about 74% of the total global methane production, up to 25% of which is consumed by anaerobic oxidation of methane (AOM). Methyl coenzyme M reductase (MCR) is the key enzyme in both methanogenesis and AOM. MCR is assembled as a dimer of two heterotrimers, where posttranslational modifications and F 430 cofactors are embedded in the active sites. However, this complex assembly process remains unknown. Here, we established a heterologous expression system for MCR to learn how MCR is assembled.

The bacterium responsible for causing tuberculosis has evolved resistance to antibiotics used to treat the disease, resulting in new multidrug resistant Mycobacterium tuberculosis (MDR-TB) and extensively drug resistant M. tuberculosis (XDR-TB) strains. Analytical techniques (1)H and (13)C Nuclear Magnetic Resonance (NMR), Fourier Transform-Ion Cyclotron Resonance with Electrospray Ionization (FT-ICR/ESI), and Matrix Assisted Laser Desorption Ionization-Mass Spectrometry (MALDI-TOF-MS) were used to study different aspects of the Cu(II)-polyethylene glycol (PEG-3350)-sucrose-isoniazid and Cu(II)-polyethylene glycol (PEG3350)-glucose-isoniazid complexes. The Cu(II) cation, sucrose or glucose, and the aggregate formed by PEG primarily serve as a composite drug delivery agent for the frontline antibiotic, however the improvement in MIC values produced with the CU-PEG-SUC-INH complex suggest an additional effect. Several Cu-PEG-SUC-INH complex variations were tested against INH resistant and nonresistant strains of M. tuberculosis.

The bacterium responsible for tuberculosis is increasing its resistance to antibiotics resulting in new multidrug-resistant Mycobacterium tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB). In this study, several analytical techniques including NMR, FT-ICR, MALDI-MS, LC-MS and UV/Vis are used to study the copper-Rifampicin-Polyethylene glycol (PEG-3350) complex. The copper (II) cation is a carrier for the antibiotic Rifampicin as well as nutrients for the bacterium. The NIH-NIAID cell line containing several Tb strains (including antibiotic resistant strains) is tested against seven copper-PEG-RIF complex variations.

We aimed to determine the effect of encapsulation on the release properties of blueberry extracts during simulated gastrointestinal digestion. An ethanolic pomace extract was microencapsulated with whey protein isolate via spray drying. The in vitro release of monomeric anthocyanins, phenolics and ferric reducing antioxidant activity of the microcapsules (W) were evaluated for the microcapsules and two non-encapsulated systems: ethanolic pomace extract (P) and freeze-dried juice (F). Concentrations of anthocyanin and phenolics were normalised prior to digestion. Results showed that antioxidant activity was in the order of: F>W>P. Regardless of encapsulation, more phenolics were released from W and P than F. Anthocyanin concentration decreased after intestinal digestion for W, but remained constant for P and F. MALDI-MS showed similar spectra for P and F but not for W. The spray-dried product has comparable release characteristics to freeze-dried juice, and may be investigated for food applications.

The copper(II) cation, sucrose, and hydroxychloroquine were complexed with the chemotherapy agent paclitaxel and studied for medicinal activity. Data (GI50, LD50) from single dose and five dose National Cancer Institute sixty cell line panels are presented. Analytical measurements of different complexes were made using Nuclear Magnetic Resonance (1H NMR), Matrix Assisted Laser Desorption Ionization-Time of Flight-Mass Spectrometry (MALDI-TOF-MS) and Fourier Transform-Ion Cyclotron Resonance (FT-ICR). Molecular modeling is utilized to better understand the impact that species could have on physical parameters associated with Lipinski's Rule of Five, such as logP and TPSA. On average, Cu(II) and hydroxychloroquine decreased GI50 values, while sucrose increased GI50 values of paclitaxel.

While there is a growing body of knowledge on disability sport consumer behavior ( Byon et al., 2011 , Byon et al., 2010a ), these studies used scales explicitly designed for non-disability sport contexts, showing only reasonable model fit and not examining factors specific to the disability sport consumer experience. This publication represents the first attempt to identify specific disability sport motives and develop a scale, the Motivation Scale for Disability Sport Consumption (MSDSC). Newly identified disability sport motives include inspiration, supercrip image and disability cultural education. These were examined in conjunction with factors from Trail and James (2001) and Trail (2010) . Data were collected at the 2011 collegiate wheelchair basketball championships; results were collected to conduct exploratory and confirmatory factor analyses. Results indicated that a 9-factor model was most appropriate. Significant motives included physical attraction, drama, escape, inspiration, physical skill, social interaction, violence, and supercrip image. The model fit was improved over the Byon studies and was comparable to relevant non-adaptive motive studies ( Lee et al., 2009 , Trail and James, 2001 , Robinson et al., 2004 ). This scale represents a tool for practitioners and academics to effectively examine spectators of disability sport.

In the first phase of this study, the binding of hydroxychloroquine to the copper(II) cation is examined using liquid chromatography-mass spectrometry (LC-MS), matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS), Fourier transform-ion cyclotron resonance spectrometry (FT-ICR) and nuclear magnetic resonance ((1)H and (13)C NMR) in one and two dimensions. The data suggest the metal-ligand complex is a polarity adaptive molecule. In the second phase of the study, the complexes activity is tested against the National Cancer Institute's 60 cell line panel. Its anti-cancer activity is compared to quinine, Cu(II)-quinine and hydroxychloroquine. It serves as a base line for future anti-cancer complexes in which hydroxychloroquine is utilized for its ability to impact cell autophagy.

Sexual isolation, the reduced tendency to mate, is one of the reproductive barriers that prevent gene flow between different species. Various species-specific signals during courtship contribute to sexual isolation between species. Drosophila albomicans and D. nasuta are closely related species of the nasuta subgroup within the Drosophila immigrans group and are distributed in allopatry. We analyzed mating behavior and courtship as well as cuticular hydrocarbon profiles within and between species. Here, we report that these two species randomly mated with each other. We did not observe any sexual isolation between species or between strains within species by multiple-choice tests. Significant difference in the courtship index was detected between these two species, but males and females of both species showed no discrimination against heterospecific partners. Significant quantitative variations in cuticular hydrocarbons between these two species were also found, but the cuticular hydrocarbons appear to play a negligible role in both courtship and sexual isolation between these two species. In contrast to the evident postzygotic isolation, the lack of sexual isolation between these two species suggests that the evolution of premating isolation may lag behind that of the intergenomic incompatibility, which might be driven by intragenomic conflicts.

A nomenclature system is presented that can be applied to a range of spherical, tubular and planar structures. The starting point in this system is determined using semi-empirical calculations that establish the atomic charge of all species. This method accommodates all elements, different substitution patterns (i.e., endo and exohedral in spherical and tubular structures) and planar structures that represent the border between molecules and materials. Many of these structures currently exist in the forms of fullerenes, carbon nanotubes and graphene structures, but the method is not limited to carbon atoms. This nomenclature system is based on the numerical position of an atom or functional group in body of the structure or its substitution pattern. Several structures including a fullerene (C72), substituted aza-fullerene, a functionalized graphene sheet and a spherical boron nitride structure are used to demonstrate the utility of the new nomenclature system.

Bryostatin-1 is a marine natural product that has demonstrated medicinal activity in pre-clinical and clinical trials for the treatment of cancer, Alzheimer's disease, effects of stroke, and HIV. In this study, iron-bryostatin-1 was obtained using a pharmaceutical aquaculture technique developed by our lab that cultivates marine bacteria for marine natural product extraction. Analytical measurements (1)H and (13)C NMR, mass spectrometry, and flame atomic absorption were utilized to confirm the presence of an iron-bryostatin-1 complex. The iron-bryostatin-1 complex produced was then tested against the National Cancer Institute's 60 cell line panel. Adding iron to bryostatin-1 lowered the anti-cancer efficacy of the compound.

In this paper, it is reported that microwave synthesis techniques can be used to convert graphite powder and fluorinated graphite in different matrices to fullerenes (i.e., C60 and C70). The microwave approach allows for the quick, economical production of the carbon allotropes. Matrix Assisted Laser Desorption Ionization Time of Flight Mass (MALDI-TOF) spectrometry is used as the detection technique. Pyrrole was also used as a reactant and the formation of aza-fullerenes was suggested by the mass spectral data.

Strawberry production in Western Australia (W.A.) is severely compromised by crown and root diseases. A range of fungi and oomycetes, including Fusarium oxysporum , binucleate Rhizoctonia , Cylindrocarpon destructans , Phoma exigua , Gnomonia fructicola , Phytophthora cactorum , Pythium ultimum and Macrophomina phaseolina , are associated with such disease outbreaks. Studies were conducted for the first time to determine and compare the virulence of the fungal and oomycete isolates, and to determine the effect of temperature on virulence of these isolates in terms of disease severity on strawberry plants. We firstly determined the virulence of these isolates at 22 °C, F. oxysporum severely affected crown, crown vascular and root tissues, resulting in the rapid wilting and death of strawberry plants; binucleate Rhizoctonia , particularly AG-A, also caused severe disease on roots and crowns, resulting in the eventual death of plants; C. destructans , P. exigua , G. fructicola , P. cactorum and P. ultimum caused variable levels of symptoms on crowns and roots, and while generally less virulent compared with F. oxysporum and Rhizoctonia , still severely retarded the growth and development of inoculated plants. Subsequent studies showed that F. oxysporum , Rhizoctonia and M. phaseolina were much more virulent under warmer conditions (>22 °C), evident as more severe disease. F. oxysporum and Rhizoctonia were most virulent and caused most severe disease at 27 °C, but M. phaseolina was most virulent and caused most severe disease at 32 °C. This relates to the dominances of these pathogens in the warmer months of the year and, in particular, the dominance of M. phaseolina in the late spring/early summer when temperatures often exceed 30 °C. This is the first study to demonstrate that F. oxysporum , binucleate Rhizoctonia and M. phaseolina are the most virulent pathogens causing crown and root diseases of strawberry in W.A., and to show that the virulence of these pathogens is influenced by the prevailing seasonal temperature regime.

Bryostatin structures share a commonality of a central bryophan ring, but each differs due to two groups (R(1) and R(2)) that are attached to the bryophan ring via ester bonds. This research examines the impact that conditions such as UV light, acidic and basic conditions can have on the bryostatin structure in the presence of octanoic acid and water. Mass spectrometry (MS) measurements suggest that bryostatin can easily rearrange into various structures under natural conditions by reacting with carboxylates that are ubiquitous in nature. A second set of measurements suggest bryostatin can be hydrolyzed by water, a reaction that has significant implications in both medicinal applications and extraction procedures.

The bacterium responsible for causing tuberculosis is increasing its resistance to antibiotics resulting in new multidrug-resistant Mycobacterium tuberculosis (MDR-TB) and extensively drug-resistant M. tuberculosis (XDR-TB) strains. In this study, several analytical techniques including NMR, FT-ICR, MALDI-MS, and LC–MS are used to study different aspects of the Copper–polyethylene glycol (PEG)–Amikacin complex. The Cu(II) cation and the aggregate formed by PEG serve as a carrier for the antibiotic. Several Cu–PEG–Amikacin complex variations were tested against NIH-NIAID cell lines containing both resistant and nonresistant strains of M. tuberculosis.

The impact of the oxidizing agent ozone (O3) on the structure of the most common spherical allotrope of carbon, buckminsterfullerene (C60) is studied. Past studies measured ozonation of C60 over short periods of time. In this study, we establish that C60 is completely degraded, not polymerized, during ozonation. The techniques, laser desorption time-of-flight mass spectrometry, UV/Vis absorbance spectroscopy, and FT-Raman are used to study the molecular degradation of the spherical allotrope C60 by O3.

Marine Natural Products (MNPs), such as bryostatin 1, are exposed to a range of physical and chemical conditions through the life cycle of the host organism. These include exposure to sunlight, oxidizing and reducing agents, cation binding, and adsorption to reactive metal oxide surfaces. Using Fourier Transform-Ion Cyclotron Resonance (FT-ICR), Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI-MS), UV/Vis absorbance spectroscopy, and molecular modeling, we studied the impact of UV light, TiO2, I2, and reaction with FeCl3 on the structure of bryostatin 1. Our results demonstrate that natural conditions transform bryostatin to a number of structures, including one with a molar mass of 806 Da, which we have previously identified in the sediment collected from the Gulf of Mexico. To date, at least 20 different structures of bryostatin have been reported in the literature. This work suggests that these variations may be products of the chemical environment in which the bryozoa Bugula neritina resides and are not the result of genetic variations within Bugula.

A discharge is arced to different solutions containing the simple molecular species water, methanol, and ammonia. We show that the impact of the discharge at the discharge-solution interface produces a range of organic molecules, including amino acids and polymers containing carboxylate, amine, imine, and cyano groups. The classic Miller-Urey experiment, of which there are hundreds of variations, changing parameters such as gas composition, pressures, and voltage have been tested, involves the production of simple amino acids and other species in the gas phase in an arc discharge. This work’s emphasis is the production of various chemical species at a discharge-liquid interface. The analysis of the product is conducted by IR, UV–VIS, LC–MS and MALDI–MS. As a laboratory exercise or a demonstration, this simple derivation of the Miller-Urey experiment can be used in a variety of teaching settings, from high school through advanced undergraduate research classes to demonstrate the basic hypothesis of how life on Earth may have started.

The innate immune system of insects responds to wounding and pathogens by mobilizing multiple pathways that provide both systemic and localized protection. Key localized responses in hemolymph include melanization, coagulation, and hemocyte encapsulation, which synergistically seal wounds and envelop and destroy pathogens. To be effective, these pathways require a targeted deposition of their components to provide protection without compromising the host. Extensive research has identified a large number of the effectors that comprise these responses, but questions remain regarding their post-translational processing, function, and targeting. Here, we used mass spectrometry to demonstrate the integration of pathogen recognition proteins, coagulants, and melanization components into stable, high-mass, multi-functional Immune Complexes (ICs) in Bombyx mori and Aedes aegypti. Essential proteins common to both include phenoloxidases, apolipophorins, serine protease homologs, and a serine protease that promotes hemocyte recruitment through cytokine activation. Pattern recognition proteins included C-type Lectins in B. mori, while A. aegypti contained a protein homologous to Plasmodium-resistant LRIM1 from Anopheles gambiae. We also found that the B. mori IC is stabilized by extensive transglutaminase-catalyzed cross-linking of multiple components. The melanization inhibitor Egf1.0, from the parasitoid wasp Microplitis demolitor, blocked inclusion of specific components into the IC and also inhibited transglutaminase activity. Our results show how coagulants, melanization components, and hemocytes can be recruited to a wound surface or pathogen, provide insight into the mechanism by which a parasitoid evades this immune response, and suggest that insects as diverse as Lepidoptera and Diptera utilize similar defensive mechanisms.

Many colleges and universities today offer students, faculty, alumni, and community members access to on-campus recreation centers. These centers offer cardiovascular and weight-training equipment, swimming pools, snack areas and juice bars, multipurpose gymnasiums, racquetball courts, locker rooms, saunas, steam rooms, and a variety of other amenities. These services are often included in a student's semester activity fee, while many institutions make the facilities available at a nominal charge for faculty and staff. In order to entice faculty and staff utilize these facilities, some institutions have begun creating and offering incentive packages for users. This research is focused on reasons why employee participation is beneficial to both the individual and the institution, how employees can be motivated through the use of incentives, the various types of participation incentives offered, examples of successful incentives, and recommendations for future incentive plans.

In recent years, the bacterium responsible for tuberculosis has been increasing its resistance to antibiotics resulting in new multidrug resistant Mycobacterium tuberculosis (MR-TB) and extensively drug-resistant tuberculosis (XDR-TB). In this study we use several analytical techniques including NMR, FT-ICR, TOF-MS, LC-MS and UV/Vis to study the copper-capreomycin complex. The copper (II) cation is used as a carrier for the antibiotic capreomycin. Once this structure was studied using NMR, FT-ICR, and MALDI-TOF-MS, the NIH-NIAID tuberculosis cell line for several Tb strains (including antibiotic resistant strains) were tested against up to seven variations of the copper-capreomycin complex. Different variations of copper improved the efficacy of capreomycin against Tb up to 250 fold against drug resistant strains of Tb.

Book review of Rick Fawn and Jeremy Larkins (eds), International Society after the Cold War: Anarchy and Order Reconsidered. London: Macmillan, 1996. 302 pp. $39.95 (paper)

A number of delivery agents, such as proteins, liposomes, micelles, and nanoparticles, are utilized for transporting pharmaceutical agents in a physiological environment. This Letter focuses on the use of the copper(II) ion and its potential role as a delivery agent for the taxanes and taxol couple to a malaria drug. Nuclear magnetic resonance (NMR, (1)H, (13)C, (15)N), Mass Spectrometry (LC-MS, MALDI-TOF, FT-ICR) and computational methods are used to examine the structure of the complex. The National Cancer Institute's benchmark 60 cell line panel is used to compare the efficacy of the copper-taxol and copper-taxol-hydroxychloroquin complexes to that of iron-taxol and pure taxol.

Hemoglobin aggregates studied under static and dynamic conditions involving the formation of nanobacteria-like structures Laser light scattering and scanning electron microscopy (SEM) are used to study hemoglobin in the aqueous phase. The impact that salts [NaCl, Ca3(PO4)2] and iron oxide nanoparticles have on the hemoglobin size are also studied. The first set of experiments examined hemoglobin aggregates in the aqueous phases in the presence of salts and nanoparticles. Aqueous phase samples were then dehydrated and examined using SEM. The resulting structures resemble those observed in nanobacteria studies conducted in other labs. This study demonstrates that aggregates of hemoglobin and various salts found in a physiological environment can produce structures that resemble nanobacteria.

Sexual isolation, the reduced ability of organisms of different species to successfully mate, is one of the reproductive barriers that prevent gene flow between different taxa. Various species-specific signals during courtship are attributed to the sexual isolation between them. Drosophila pseudoobscura has been widely used to study the behavioral and genetic mechanisms underlying selection for sexual isolation, as a model system for speciation. D. pseudoobscura and its sibling species, D. persmilis, live together in many locations but are reproductively isolated from one another. North American geographic strains of D. pseudoobscura from the American West mate at random. Several decades ago, D. pseudoobscura was collected in the vicinity of Bogota, Colombia, and later named the subspecies D. pseudoobscura bogotana. Nearly 5,000 matings were observed in this study. We analyzed mating behavior and cuticular hydrocarbon profiles as well as courtship within and between North American and Bogota strains of D. pseudoobscura. Here we report for the first time that Bogota strains of D. pseudoobscura do show statistically significant sexual isolation from North American strains. In addition, there are quantitative variations in cuticular hydrocarbons as well as in courtship behavior between Bogota and North American strains, and females of both North American and Bogota strains show strong preference for North American strain males having high mating propensities, suggesting that the Bogota strains are at an early stage that could lead to a separate species.

Iron chelators are being examined as a potential class of pharmaceutical agents to battle different types of cancer as well as iron overload diseases. In recent studies, iron binding species such as desferrioxamine, triapine, tachpyridine, Dp44Mt, and PIH have been tested in cell line tests and clinical trials. Using published chemical equilibrium values (stability constants, equilibrium constants), it is argued that an iron chelator cannot competitively remove iron from a heme-containing biomolecule (i.e. hemoglobin (Hb), myoglobin) causing a cancerous cell to die. This type of reaction (DFO(aq) + [Fe(2+,3+)-Hb] --[Fe(2+,3+)-DFO] + Hb) has been proposed in a number of published studies using circumstantial evidence. It is argued that iron chelators can potentially interact with iron from ferritin or iron that has precipitated or flocculated as oxyhydroxide under physiological pH's. It is argued that chelators can interfere with various physiological processes by binding cations such as Ca(2+), Zn(2+) or K(+). A number of siderophores and natural products that have the ability to bind Fe(3+)/Fe(2+) as well as other cations are discussed in terms of their potential pharmaceutical activity as chelators. Chemical equilibria between cations and pharmaceutical agents, which are rarely quantitated in explaining medicinal mechanisms, are used to show that chelators can bind and remove iron and other cations from physiologically important systems required for cell survival and propagation.

In this computational study, geometric factors are calculated by applying semi-empirical methods (PM3) that support experimental evidence from this lab where bryostatins can bind trivalent iron with six Fe-O bonds forming an octahedral geometry. The geometric factors are calculated for all 20 structures (Fe3+ bound to bryostatin 1-20) as a neutral, monovalent, and divalent species. The average Fe-O bond distances and bond angles are compared to those of known marine and terrestrial siderophores. From these two data sets, we then examined other known marine natural products (MNPs) that can form a hexavalent complex with six Fe-O bonds and draw conclusions about their potential biological role as marine siderophores. This computational data indicates that Fe(III) strongly bonds to a host of MNPs, increasing their water solubility, contracting their structure, hence allowing transport through cell membranes more readily, and in some cases, stabilizing ester bonds that are susceptible to hydrolysis. It is argued that administering medicinally bryostatin, its analogs or other MNPs as a ferric complex, holds some fundamental chemical advantages compared to its administration as a neutral uncomplexed species.

The removal of water-soluble sulphonated phenylazonaphthol dye effluents generated by textile industries is an important issue in wastewater treatment. Microbial treatment of environmental pollutants including dyes, with white rot fungi has received wide attention as a potential alternative for conventional methods in wastewater treatment. Three sulphonated phenylazonaphthol dyes with similar molecular structures Acid Orange 7, Acid Orange 8 and Mordant Violet 5 were selected and degraded by the white rot fungus Pleurotus ostreatus. Chemical instrumental analysis methods such as high-performance liquid chromatography (HPLC) and capillary electrophoresis combined with electrospray ionization mass spectrometry (CE-ESI-MS) were used to identify the degraded dyes. Mordant Violet 5 had two degradation pathways when degraded by P. ostreatus. The first degradation pathway for Mordant Violet 5 was for trans structure and the cis-Mordant Violet 5 followed the second pathway. Acid Orange 8 and Acid Orange 7 had the same degradation mechanism as the first degradation mechanism for Mordant Violet 5, that is cleavage of azo bond at the naphthalene ring side where benzenesulfonic acid and 1,2-naphthoquinone are formed.

Biodegradation products from four model sulfonated azo dyes Orange II, Acid Orange 8, Food Yellow 3, and 4-[(4-hydroxyphenyl)azo]-benzenesulfonic acid, sodium salt (4HABA), during fungal degradation were determined by capillary electrophoresis coupled with ion trap mass spectrometry (CE-MS) with electrospray ionization and a coaxial sheath flow interface. The development and optimization of this analytical method including the sheath liquid composition and flow rate, nebulizing gas flow rate, carrier electrolyte, and MS voltage are described herein. Detection of unknown biodegradation products was carried out under negative ion mode with base peak electrophorogram (BPE) or extractive ion electrophorogram (EIE) monitoring. A volatile ammonium acetate buffer (10 mM) without organic modifier and a shealth liquid made from 2-propanol and water (80:20, v/v) were suited for the separation and ESI interface. The sulfonated ion was the base peak for model azo dyes and their metabolites containing sulfonic group. Results showed that the tested azo dyes were degraded quickly in the culture of white rot fungus, Pleurotus ostreatus in 3 days with the major biodegradation products being 4-hydroxy-benzenesulfonic acid, 3-methyl-4-hydroxy-benzenesulfonic acid, benzenesulfonic acid, 1,2-naphthoquinone-6-sulfonic acid and 3-methyl-benzenesulfonic acid.

The sea squirt Ecteinascidia turbinata produces the powerful drug ET743. In this study Inductively Coupled Plasma-Atomic Emission Spectroscopy (ICP-AES) and Matrix Assisted Laser Desorption Ionization-Mass Spectrometry (MALDI-MS) are systematically used to measure elemental and molecular species in a Florida Keys mangrove ecosystem that contains the sea squirt. ICP-AES is used to measure the concentration of 27 elements down to the parts per billion level in 16 organisms and 3 sediment samples that reside in the mangrove ecosystem including turtle grass, blue crabs, fire sponge, and lettuce slugs. MALDI-MS is used to search for ET743 in these same organisms and sediment samples. A mass spectral feature corresponding to ET743 is identified in the extract of the sea squirt, red mangrove root (Rhizophera mangle), the schoolmaster snapper (Lutjanus griseus), and a sediment sample taken from the ecosystem. We use MALDI-MS to study the impact that various environmental conditions, such as UV light, I(2), cation binding (Fe(+3), Zn(+2), Pb(+2), Cu(+2)), metal oxide nanoparticles (FeO, CuO, TiO(2), ZnO, Al(2)O(3)), a common mineral (CaCO(3)), and extremes in acidity (0.1 M HCl, 0.1 M NaOH) have on the ET743 structure. The data provide potential structures (degradation products, metal-ligand complexes, etc.) that might be present in organism or sedimentary extracts that are similar to ET743. We are studying the marine geochemistry of this ecosystem so a broth can be developed and tested for producing this marine natural product.

The bryozoan species Bugula neritina contains the anticancer agent bryostatin. Bryostatin has been extracted from these sessile marine invertebrates since the late 1960s from the Gulf of California, Gulf of Mexico, as well as various locations on the eastern and western rims of the Pacific Ocean. In this work we are focusing on animals harvested in the Gulf of Mexico near Alligator Point (Florida). Using Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) we measure the concentration of 70 elements in B. neritina, a sea squirt, and the sediment from the point of harvesting. This data has helped us generate an extraction process for marine natural products. Combining UV/VIS absorbance measurements with Matrix Assisted Laser Desorption Ionization-Time of Flight-Mass Spectrometer (MALDI-TOF-MS), we demonstrated that the specific form of bryostatin extracted is a function of the solvent. A 9.4T Fourier Transform-Ion Cyclotron Resonance (FT-ICR) mass spectrometer, whose sensitivity, mass accuracy, and resolving power allowed the exact empirical formulas of potential precursors of bryostatin to be identified, was employed. Finally we examine extracts of 14 marine species of the Gulf of Mexico, from the sand trout (Cynoscion arenarius) to chicken liver sponge (Chrondrilla nucula), all recently collected, which had shown some medicinal activity thirty years ago in a National Cancer Institute study. By the MALDI-TOF-MS, we were able to identify mass spectral features that correspond to different variations of the basic bryostatin structure, which raises the question if the bryozoans are the original source of bryostatin.