Toxic aspergilli from pistachio nuts

Pistachio nut samples taken during various stages of development from orchards in Iran, showed that contamination with fungi occurred mainly during the later stages of nut development. Members of the genera Aspergillus and Penicillium occurred most frequently. Of the Aspergilli, the species A. niger, A. flavus and A. fischeri var. spinosus occurred most frequently, followed by A. terreus, A. tamarii and A. nidulans. Twenty-two isolates comprising 13 species were tested for toxicity to ducklings. Isolates of known toxic fungi included A. flavus, A. niger, A. parasiticus, A. ochraceus, A. versicolor, A. nidulans and A. terreus. The toxicity of A. fischeri var. spinosus is reported. Chemical analysis showed that all isolates of A. flavus and A. parasiticus produced aflatoxin B₁, the isolates of A. versicolor and A. nidulans produced sterigmatocystin while the toxic isolate of A. ochraceus did not produce ochratoxins.Toxic fungi have been shown to occur in a variety of nuts (4), (5), (11), (12), (13), (15), (18), (20), (21). Aflatoxin contamination of pistachio nuts has been reported and has in the past led to the rejection of consignments of Iranian pistachio nuts (1). In Iran, pistachio nuts are produced mainly in the south eastern provinces (Kerman & Zahedan) and to a limited extent in the Northern part (Kazvin & Damghan). In 1975 it was estimated (7) that there were some 24 million pistachio trees in Iran, of which 60% were situated in Rafsanjan, Kerman (Table 1). Economic considerations as well as the potential health hazard posed by aflatoxin-contaminated nuts, prompted the University of Isfahan to initiate a study of various aspects of the mycotoxin problem in pistachio nuts.     

Lakeside reed border characteristics at Lake Constance (Untersee): A comparison between 1981–1983 and 1994

Eutrophication and the associated proliferation offilamentous algae were often considered importantcauses of reed die-back that occurred in the sixtiesand seventies at Lake Constance. According to thishypothesis the reduction in lake water nutrientconcentrations during the eighties and nineties shouldhave reduced the abundance of filamentous algae andwhich should have reduced the supposed mechanicalimpact of filamentous algae on the lakeside reedstands. To examine whether this had actually occurred,the condition of the lakeside reed border at LakeConstance (Untersee) was surveyed in 1994 and resultswere compared to those of similar surveys carried outin the years 1981–83. The extent of mechanical damageto reed stalks, the amount of macrophyte wash and thedegree of shoreline erosion were assessed forshoreline segments of 25 m along a total shorelinelength of 65 km. The results showed that mechanicaldamage to reed stalks was generally higher in 1994than in 1981–83, while macrophyte wash and shorelineerosion were generally less severe. The decreasedamount of macrophyte wash was associated with a changein macrophyte species composition from a dominance offloating filamentous algae to a dominance of rootedPotamogetonaceaeand fragile Characeae,and therefore probably was due to the lower nutrientconcentrations of the lake. The opposite trends foundfor mechanical damage to reed stands and the amount ofmacrophyte wash between 1981–83 and 1994, as well asthe absence of relation between the two variables in1994, suggested that macrophyte wash had only a minoreffect on the condition of lakeside reed stands atLake Constance. Highly damaged reed stands and erodedcliffs were often located at wind exposed sites orclose to highly frequented shipping routes in 1994,suggesting that mechanical stress from waves wasresponsible for the damage. Due to the lakewardprogression of reed stands during the early nineties,reed stalks were, on average, located in deeper waterin 1994 than in 1981–83. This might have strengthenedthe mechanical stress from wave action even more andhave caused the higher frequency of mechanical damageto reed stalks. Other causes of damage such asmacrophyte wash, roosts of birds or bird swarmsprobably played a minor and only local role.     

An Improved Protocol for Quantification of Freshwater Actinobacteria by Fluorescence In Situ Hybridization

We tested a previously described protocol for fluorescence in situ hybridization of marine bacterioplankton with horseradish peroxidase-labeled rRNA-targeted oligonucleotide probes and catalyzed reporter deposition (CARD-FISH) in plankton samples from different lakes. The fraction of Bacteria detected by CARD-FISH was significantly lower than after FISH with fluorescently monolabeled probes. In particular, the abundances of aquatic Actinobacteria were significantly underestimated. We thus developed a combined fixation and permeabilization protocol for CARD-FISH of freshwater samples. Enzymatic pretreatment of fixed cells was optimized for the controlled digestion of gram-positive cell walls without causing overall cell loss. Incubations with high concentrations of lysozyme (10 mg ml⁻¹) followed by achromopeptidase (60 U ml⁻¹) successfully permeabilized cell walls of Actinobacteria for subsequent CARD-FISH both in enrichment cultures and environmental samples. Between 72 and >99% (mean, 86%) of all Bacteria could be visualized with the improved assay in surface waters of four lakes. For freshwater samples, our method is thus superior to the CARD-FISH protocol for marine Bacteria (mean, 55%) and to FISH with directly fluorochrome labeled probes (mean, 67%). Actinobacterial abundances in the studied systems, as detected by the optimized protocol, ranged from 32 to >55% (mean, 45%). Our findings confirm that members of this lineage are among the numerically most important Bacteria of freshwater picoplankton.     

Tamoxifen and flaxseed alter angiogenesis regulators in normal human breast tissue in vivo

The incidence of breast cancer is increasing in the Western world and there is an urgent need for studies of the mechanisms of sex steroids in order to develop novel preventive strategies. Diet modifications may be among the means for breast cancer prevention. Angiogenesis, key in tumor progression, is regulated by the balance between pro- and anti-angiogenic factors, which are controlled in the extracellular space. Sampling of these molecules at their bioactive compartment is therefore needed. The aims of this study were to explore if tamoxifen, one of the most used anti-estrogen treatments for breast cancer affected some of the most important endogenous angiogenesis regulators, vascular endothelial growth factor (VEGF), angiogenin, and endostatin in normal breast tissue in vivo and if a diet supplementation with flaxseed had similar effects as tamoxifen in the breast. Microdialysis was used for in situ sampling of extracellular proteins in normal breast tissue of women before and after six weeks of tamoxifen treatment or before and after addition of 25 g/day of ground flaxseed to the diet or in control women. We show significant correlations between estradiol and levels of VEGF, angiogenin, and endostatin in vivo, which was verified in ex vivo breast tissue culture. Moreover, tamoxifen decreased the levels of VEGF and angiogenin in the breast whereas endostatin increased significantly. Flaxseed did not alter VEGF or angiogenin levels but similar to tamoxifen the levels of endostatin increased significantly. We conclude that one of the mechanisms of tamoxifen in normal breast tissue include tipping of the angiogenic balance into an anti-angiogenic state and that flaxseed has limited effects on the pro-angiogenic factors whereas the anti-angiogenic endostatin may be modified by diet. Further studies of diet modifications for breast cancer prevention are warranted.     

Peripheral Blood Neutrophilia as a Biomarker of Ozone-Induced Pulmonary Inflammation

Background

Ozone concentrations are predicted to increase over the next 50 years due to global warming and the increased release of precursor chemicals. It is therefore urgent that good, reliable biomarkers are available to quantify the toxicity of this pollutant gas at the population level. Such a biomarker would need to be easily performed, reproducible, economically viable, and reflective of ongoing pathological processes occurring within the lung.

Methodology

We examined whether blood neutrophilia occurred following a controlled ozone challenge and addressed whether this could serve as a biomarker for ozone-induced airway inflammation. Three separate groups of healthy subjects were exposed to ozone (0.2 ppm, 2h) and filtered air (FA) on two separate occasions. Peripheral blood samples were collected and bronchoscopy with biopsy sampling and lavages was performed at 1.5h post exposures in group 1 (n=13), at 6h in group 2 (n=15) and at 18h in group 3 (n=15). Total and differential cell counts were assessed in blood, bronchial tissue and airway lavages.

Results

In peripheral blood, we observed fewer neutrophils 1.5h after ozone compared with the parallel air exposure (-1.1±1.0x10⁹ cells/L, p<0.01), at 6h neutrophil numbers were increased compared to FA (+1.2±1.3x10⁹ cells/L, p<0.01), and at 18h this response had fully attenuated. Ozone induced a peak in neutrophil numbers at 6h post exposure in all compartments examined, with a positive correlation between the response in blood and bronchial biopsies.

Conclusions

These data demonstrate a systemic neutrophilia in healthy subjects following an acute ozone exposure, which mirrors the inflammatory response in the lung mucosa and lumen. This relationship suggests that blood neutrophilia could be used as a relatively simple functional biomarker for the effect of ozone on the lung.     

Control of endogenous gene expression timing by introns

Comparison of gene expression from transgenes and endogenous genes with or without introns reveals a time-regulating role of introns in natural biological systems.     

Transthyretin Interferes with Aβ Amyloid Formation by Redirecting Oligomeric Nuclei into Non-Amyloid Aggregates

The pathological Aβ aggregates associated with Alzheimer's disease follow a nucleation-dependent path of formation. A nucleus represents an oligomeric assembly of Aβ peptides that acts as a template for subsequent incorporation of monomers to form a fibrillar structure. Nuclei can form de novo or via surface-catalyzed secondary nucleation, and the combined rates of elongation and nucleation control the overall rate of fibril formation. Transthyretin (TTR) obstructs Aβ fibril formation in favor of alternative non-fibrillar assemblies, but the mechanism behind this activity is not fully understood. This study shows that TTR does not significantly disturb fibril elongation; rather, it effectively interferes with the formation of oligomeric nuclei. We demonstrate that this interference can be modulated by altering the relative contribution of elongation and nucleation, and we show how TTR's effects can range from being essentially ineffective to almost complete inhibition of fibril formation without changing the concentration of TTR or monomeric Aβ.