Bioconversion of Aflatoxin G1 to Aflatoxin B3 by selected fungi

Aflatoxin G₁ (AFG₁) was transformed into aflatoxin B₃ (AFB₃) by the fungiRhodotorula sp,Sporobolomyces sp,Rhizopus oryzae NRRL395,Pythium ultimum, Aspergillus terreus, A clavatus and Penicillium frequentans grown in a medium containing AFG₁, Difco potato dextrose broth, yeast extract, and peptone both in liquid shaken cultures and in solid static cultures at 25°C in the dark. A maximum rate of transformation of 10 % was obtained after 2 to 3 weeks of incubation. The transformation was correlated with an increase in the pH of the media from 5.7 –5.9 to 8.3 – 8.8.Saccharomyces cerevisiae also transformed AFG₁ into AFB₃, but at a slower rate; the pH of the media did not reach above 8.0 until 5 weeks after incubation. No transformation was observed whenA niger andP chrysogenum were tested; in both cases, no increase in pH was noticed. However, some transformation of AFG₁ to AFB₃ by both fungi was observed when the initial pH of the media was adjusted to 9.0. The rate of transformation increased to 15 – 20% in the static culture where the same medium was adsorbed onto vermiculite andRhizopus andAspergilli gave the highest increase in AFB₃ yield.     

Influence of solute and hydrogen ion concentration on the water relations of some xerophilic fungi.

Germination and growth of six xerophilic fungi, Aspergillus flavus, Aspergillus ochraceus, Eurotium chevalieri, Chrysosphorium fastidium, Wallemia sebi and Xeromyces bisporus were examined on media of a wide range of water activities (aw). The influence of three solutes, NaCl, glycerol and a glucose/fructose mixture, was studied at pH 4-0 and pH 6-5 using a plate-slide technique. Germination times and growth rates were affected by solute type, but the influence of pH was less marked. Except for Wallemia sebi, the fungi grew most strongly on glucose/fructose and were partially or completely inhibited by NaCl. The results showed that a universal isolation medium for xerophilic fungi could be based on glycerol or glucose/fructose but not on NaCl as aw-limiting solute.     

An invasive frog, Eleutherodactylus coqui, increases new leaf production and leaf litter decomposition rates through nutrient cycling in Hawaii

A frog endemic to Puerto Rico, Eleutherodactylus coqui, invaded Hawaii in the late 1980s, where it can reach densities of 50,000 individuals ha⁻¹. Effects of this introduced insectivore on invertebrate communities and ecosystem processes, such as nutrient cycling, are largely unknown. In two study sites on the Island of Hawaii, we studied the top-down effects of E. coqui on aerial, herbivorous, and leaf litter invertebrates; herbivory, plant growth, and leaf litter decomposition rates; and leaf litter and throughfall chemistry over 6 months. We found that E. coqui reduced all invertebrate communities at one of the two study sites. Across sites, E. coqui lowered herbivory rates, increased NH₄⁺ and P concentrations in throughfall, increased Mg, N, P, and K in decomposing leaf litter, increased new leaf production of Psidium cattleianum, and increased leaf litter decomposition rates of Metrosideros polymorpha. In summary, E. coqui effects on invertebrates differed by site, but E. coqui effects on ecosystem processes were similar across sites. Path analyses suggest that E. coqui increased the number of new P. cattleianum leaves and leaf litter decomposition rates of M. polymorpha by making nutrients more available to plants and microbes rather than through changes in the invertebrate community. Results suggest that E. coqui in Hawaii has the potential to reduce endemic invertebrates and increase nutrient cycling rates, which may confer a competitive advantage to invasive plants in an ecosystem where native species have evolved in nutrient-poor conditions.     

Glutamate Mediated Astrocytic Filtering of Neuronal Activity

Neuron-astrocyte communication is an important regulatory mechanism in various brain functions but its complexity and role are yet to be fully understood. In particular, the temporal pattern of astrocyte response to neuronal firing has not been fully characterized. Here, we used neuron-astrocyte cultures on multi-electrode arrays coupled to Ca²⁺ imaging and explored the range of neuronal stimulation frequencies while keeping constant the amount of stimulation. Our results reveal that astrocytes specifically respond to the frequency of neuronal stimulation by intracellular Ca²⁺ transients, with a clear onset of astrocytic activation at neuron firing rates around 3-5 Hz. The cell-to-cell heterogeneity of the astrocyte Ca²⁺ response was however large and increasing with stimulation frequency. Astrocytic activation by neurons was abolished with antagonists of type I metabotropic glutamate receptor, validating the glutamate-dependence of this neuron-to-astrocyte pathway. Using a realistic biophysical model of glutamate-based intracellular calcium signaling in astrocytes, we suggest that the stepwise response is due to the supralinear dynamics of intracellular IP₃ and that the heterogeneity of the responses may be due to the heterogeneity of the astrocyte-to-astrocyte couplings via gap junction channels. Therefore our results present astrocyte intracellular Ca²⁺ activity as a nonlinear integrator of glutamate-dependent neuronal activity.     

Vascular gene transfer of the human inducible nitric oxide synthase: characterization of activity and effects on myointimal hyperplasia.

BACKGROUND: Nitric oxide (NO) has been shown to decrease myointimal hyperplasia in injured blood vessels. We hypothesize inducible No synthase (iNOS) gene transfer even at low efficiency will provide adequate local no production to achieve this goal. MATERIALS AND METHODS: A retroviral vector containing the human iNOS cDNA (DFGiNOS) was used to transfer the iNOS gene into vascular cells and isolated blood vessels to answer the following questions: can vascular endothelial and smooth muscle cells support iNOS activity and will low efficiency iNOS gene transfer suppress myointimal hyperplasia in injured porcine arteries? RESULTS: DFGiNOS-infected sheep pulmonary artery endothelial cells (SPAEC) expressed significant iNOS mRNA and protein, releasing nitrite levels of 155.0 +/- 10.7 nmol/mg protein/24 h vs. 5.5 +/- 1.1 by control cells. Transduced rat smooth muscle cells (RSMC) also expressed abundant iNOS mRNA and protein, but, in contrast to SPAEC, NO synthesis was dependent on exogenous tetrahydrobiopterin (BH4) (291.8 +/- 10.4 nmol nitrite/mg protein/24 hr with BH4, 37.7 +/- 2.6 without BH4). Only porcine arteries infected with DFGiNOS following balloon injury exhibited a 3-fold increase in total NO synthesis and a 15-fold increase in cGMP levels over control vessels in a BH4 dependent fashion, despite only a 1% gene transfer efficiency. Transfer of iNOS completely prevented the 53% increase in myointimal thickness induced by balloon catheter injury; the administration of a NOS inhibitor reversed this effect. CONCLUSIONS: These in vitro findings suggest that vascular iNOS gene transfer may be feasible. Furthermore, a low gene transfer efficiency may be sufficient to inhibit myointimal hyperplasia following arterial balloon injury, although a source of BH4 may be required.     

Oxidative lipidomics of γ-radiation-induced lung injury: mass spectrometric characterization of cardiolipin and phosphatidylserine peroxidation

Oxidative damage plays a significant role in the pathogenesis of γ-radiation-induced lung injury. Endothelium is a preferred target for early radiation-induced damage and apoptosis. Given the newly discovered role of oxidized phospholipids in apoptotic signaling, we performed oxidative lipidomics analysis of phospholipids in irradiated mouse lungs and cultured mouse lung endothelial cells. C57BL/6NHsd female mice were subjected to total-body irradiation (10 Gy, 15 Gy) and euthanized 24 h thereafter. Mouse lung endothelial cells were analyzed 48 h after γ irradiation (15 Gy). We found that radiation-induced apoptosis in vivo and in vitro was accompanied by non-random oxidation of phospholipids. Cardiolipin and phosphatidylserine were the major oxidized phospholipids, while more abundant phospholipids (phosphatidylcholine, phosphatidylethanolamine) remained non-oxidized. Electrospray ionization mass spectrometry analysis revealed the formation of cardiolipin and phosphatidylserine oxygenated molecular species in the irradiated lung and cells. Analysis of fatty acids after hydrolysis of cardiolipin and phosphatidylserine by phospholipase A(2) revealed the presence of mono-hydroperoxy and/or mono-hydroxy/mono-epoxy, mono-hydroperoxy/mono-oxo molecular species of linoleic acid. We speculate that cyt c-driven oxidations of cardiolipin and phosphatidylserine associated with the execution of apoptosis in pulmonary endothelial cells are important contributors to endothelium dysfunction in γ-radiation-induced lung injury.     

Single-molecule anisotropy imaging

A novel method, single-molecule anisotropy imaging, has been employed to simultaneously study lateral and rotational diffusion of fluorescence-labeled lipids on supported phospholipid membranes. In a fluid membrane composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, in which the rotational diffusion time is on the order of the excited-state lifetime of the fluorophore rhodamine, a rotational diffusion constant, D(rot) = 7 x 10(7) rad(2)/s, was determined. The lateral diffusion constant, measured by direct analysis of single-molecule trajectories, was D(lat) = 3.5 x 10(-8) cm(2)/s. As predicted from the free-volume model for diffusion, the results exhibit a significantly enhanced mobility on the nanosecond time scale. For membranes of DPPC lipids in the L(beta) gel phase, the slow rotational mobility permitted the direct observation of the rotation of individual molecules characterized by D(rot) = 1.2 rad(2)/s. The latter data were evaluated by a mean square angular displacement analysis. The technique developed here should prove itself profitable for imaging of conformational motions of individual proteins on the time scale of milliseconds to seconds.     

Potential consequences of the coqui frog invasion in Hawaii

The Puerto Rican frog, Eleutherodactylus coqui, has invaded Hawaii and has negatively impacted the state's multimillion dollar floriculture, nursery and tourist industries; however, little is known about the ecological consequences of the invasion. Using data from Puerto Rico and Hawaii, the authors summarize the potential consequences of the invasion and describe future research needs. It could be predicted that the coqui would reduce the abundance of Hawaii's endemic invertebrates. However, data suggest that coquis are mostly consuming non‐native invertebrates, and not invertebrate pests, such as mosquitoes and termites. Endemic invertebrates are likely to represent a portion of the coqui diet, but it remains uncertain which endemic invertebrates are most threatened by coqui predation and whether there will be indirect effects that benefit or harm them. It could be predicted that coquis would compete with endemic birds for invertebrate prey, but there is presently little overlap in the habitats used by coquis and endemic birds. Although, coquis may make bird re‐invasion into lowland ecosystems more difficult; alternatively, coquis could serve as an additional food source for some endemic birds. Finally, it could be predicted that coquis serve as a food source for endemic‐bird predators, such as rats and mongoose, and bolster their abundance. Preliminary data suggest that coquis will not bolster rat or mongoose populations. Managing coqui populations in Hawaii has been a challenge. A population has not yet been eradicated using citric acid, the only federally approved pesticide for coquis. It is unlikely that coquis will ever be eradicated from the islands of Hawaii and Maui, where there are now hundreds of populations. Quick and severe responses to new introductions may be the only effective means of containing the spread of the coqui.     

Extraction and electrospinning of gelatin from fish skin

Ultra-fine gelatin fibers were successfully fabricated by electrospinning from the solutions of Nile tilapia (Oreochromis niloticus) skin-extracted gelatin in either acetic acid or formic acid aqueous solutions. The extracted gelatin contained 7.3% moisture, 89.4% protein, 0.3% lipid, and 0.4% ash contents (on the basis of wet weight), while the bloom gel strength, the shear viscosity, and the pH values were 328 g, 17.8 mPa s, and 5.0, respectively. Both the acid concentration and the concentration of the gelatin solutions strongly influenced the properties of the as-prepared solutions and the obtained gelatin fibers. At low acid concentrations (i.e., 15% (w/v) extracted gelatin solutions in 10 and 20% (v/v) acetic acid solvents or 10-60% (v/v) formic acid solvents), a combination between smooth and beaded fibers was observed. At low concentrations of the gelatin solutions in either 40% (v/v) acetic acid solvent or 80% (v/v) formic acid solvent (i.e., 5-11%, w/v), either discrete beads or beaded fibers were obtained, while, at higher concentrations (i.e., 14-29%, w/v), only smooth or a combination of smooth and beaded fibers were obtained. The average diameters of the obtained fibers, regardless of the types of the acid solvents used, ranged between 109 and 761 nm. Lastly, cross-linking of the obtained gelatin fiber mats with glutaraldehyde vapor caused slight shrinkage from their original dimension, and the cross-linked gelatin fiber mats became stiffer.