Atrazine-induced degranulation of thyroid mast cells in peripubertal and adult rats

Atrazine is a commonly used pesticide in the US and the non-EU countries. It is classified as an endocrine-disrupting chemical and is well-known for its reproductive toxicity in mammals and lower vertebrates. The study on atrazine effects on thyroid mast cells was performed on juvenile/peripubertal and adult male Wistar rats orally gavaged with atrazine at doses of 50 mg/kg of body weight (bw) or 200 mg/kg bw. In order to visualize the mast cell population within the thyroid gland, a histochemical staining method of toluidine blue was used. The results of the histological evaluation demonstrated a prominent increase in mast cell degranulation in both age groups and at both atrazine doses. According to the stereological analysis, a statistically significant decrease in the mast cell volume density in the young rats exposed to a higher dose of atrazine was found when compared to the corresponding control. The numerical density of mast cells significantly decreased in a higher-dose atrazine treated adults in comparison to the control. The obtained data suggest that atrazine-affected mast cells would probably have a consequent influence on thyroid follicular cells and/or thyroid microvasculature via paracrine action of released mediators, but might also be involved in already suggested thyroid cancerogenesis.     

Radioprotectors – the Evergreen Topic

To protect organisms from ionizing radiation (IR), and to reduce morbidity or mortality, various agents, called radioprotectors, have been utilized. Because radiation‐induced cellular damage is attributed primarily to the harmful effects of free radicals, molecules with radical‐scavenging properties are particularly promising as radioprotectors. Early development of such agents focused on thiol synthetic compounds, known as WR protectors, but only amifostine (WR‐2721) has been used in clinical trials as an officially approved radioprotector. Besides thiol compounds, various compounds with different chemical structure were investigated, but an ideal radioprotector has not been found yet. Plants and natural products have been evaluated as promising sources of radioprotectors because of their low toxicity, although they exhibit an inferior protection level compared to synthetic thiol compounds. Active plant constituents seem to exert the radioprotection through antioxidant and free radical‐scavenging activities. Our research established that plants containing polyphenolic compounds (raspberry, blueberry, strawberry, grape, etc.) exhibit antioxidative activities and protect genetic material from IR.     

Protective activity of (1S,2E,4R,6R,7E,11E)-2,7,11-cembratriene-4,6-diol analogues against diisopropylfluorophosphate neurotoxicity: Preliminary structure–activity relationship and pharmacophore modeling

Diisopropylfluorophosphate (DFP) is an organophosphorous insecticide used as a surrogate for the more toxic chemical warfare nerve agent sarin. DFP produces neurotoxicity in vivo and irreversibly decreases the area of population spikes recorded from the CA1 region of acute hippocampal slices. (1S,2E,4R,6R,7E,11E)-2,7,11-Cembratriene-4,6-diol (1) is a neuroprotective natural cembranoid that reverses DFP-induced damage both in vivo and in the hippocampal slice. Cembranoid 1 acts by noncompetitive inhibition of the α7 nicotinic acetylcholine receptor. This study aims at establishing a preliminary structure–activity relationship to define the neuroprotective cembranoid pharmacophores using the hippocampal slice assay and pharmacophore modeling. Fourteen natural, semisynthetic, or biocatalytic cembranoid analogues 2–15 related to 1 were tested for their capacity to protect the population spikes from DFP-induced damage and intrinsic toxicity. Twelve cembranoids caused significant reversal of DFP toxicity; only 3 active analogues displayed minor intrinsic toxicity at 10 μM. The C-4 epimer of 1 (2) and the 4-O-methyl ether analogue of 1 (3), were totally devoid of neuroprotective activity. The results suggested a model for cembranoid binding where the hydrophobic ring surface binds to a hydrophobic (Hbic) patch on the receptor molecule and an electronegative atom (oxygen or sulfur) in proper spatial relationship to the ring surface interacts with an electropositive group in the receptor binding site. A pharmacophore model consisting of 1 hydrogen bond acceptor (HBA), 2 Hbic, and 10 exclusion spheres was established using HipHop-REFINE and supported the above mentioned pharmacophoric hypothesis.     

Surface structure of the two porcine low-density lipoprotein subclasses — a spin labelling study

Porcine low-density lipoprotein (LDL) subclasses were spin-labelled at both protein and lipid sites. The surface structures of the two subclasses were compared. E.s.r. spectra of stearic acid spin-labels [I(m/n)] and of androstanol spin-label indicated more restricted motion of the labels in LDL₂ (d = 1.063?1.090 g/ml) than in LDL₁ (d = 1.020?1.063 g/ml). Thermotropic change in the surface structure was found in both subclasses by both protein and lipid spin-labels, but at lower temperature in LDl₁, than in LDL₂. These results indicate the relationship between the size and the dynamics of the lipid components in the surface layer of the LDL subclasses.     

Temperature-dependent conformation change in spin-labeled hemoglobin

Hemoglobin was spin labeled at β-93(F9)-cysteine with N-oxy-2,2,6,6-tetramelhylpiperidinylmaleimide. The inward shift of the high-field hyperfine line (ΔH_XXX) position in the ESR spectra of the Spin label was measured aS a function of temperature. One can expect that an abrupt change in the microenvironment around the tightly bound spin label will be reflected in the function ΔH_XXX(T) as a discontinuity (break point). This was shown for aquo-, azido-. nitro- and oxyhemoglobin derivatives. The presented results suggest that the microenvironment around the tightly hound spin label in those methemoglobin derivatives that exhibit the mixed-spin state of the heme iron is prone to an abrupt change above a certain ligand-specific temperature. The change in microenvironment of the spin label is probably due to a temperature-dependent change in the tertiary structure of the protein.     

Adhesion of Acinetobacter venetianus to Diesel Fuel Droplets Studied with In Situ Electrochemical and Molecular Probes

The adhesion of a recently described species, Acinetobacter venetianus VE-C3 (F. Di Cello, M. Pepi, F. Baldi, and R. Fani, Res. Microbiol. 148:237–249, 1997), to diesel fuel (a mixture of C₁₂ to C₂₈ n-alkanes) and n-hexadecane was studied and compared to that of Acinetobacter sp. strain RAG-1, which is known to excrete the emulsifying lipopolysaccharide, emulsan. Oxygen consumption rates, biomass, cell hydrophobicity, electrophoretic mobility, and zeta potential were measured for the two strains. The dropping-mercury electrode (DME) was used as an in situ adhesion sensor. In seawater, RAG-1 was hydrophobic, with an electrophoretic mobility (μ) of −0.38 × 10⁻⁸ m² V⁻¹ s⁻¹ and zeta potential (ζ) of −4.9 mV, while VE-C3 was hydrophilic, with μ of −0.81 × 10⁻⁸ m² V⁻¹ s⁻¹ and ζ of −10.5 mV. The microbial adhesion to hydrocarbon (MATH) test showed that RAG-1 was always hydrophobic whereas the hydrophilic VE-C3 strain became hydrophobic only after exposure to n-alkanes. Adhesion of VE-C3 cells to diesel fuel was partly due to the production of capsular polysaccharides (CPS), which were stained with the lectin concanavalin A (ConA) conjugated to fluorescein isothiocyanate and observed in situ by confocal microscopy. The emulsan from RAG-1, which was negative to ConA, was stained with Nile Red fluorochrome instead. Confocal microscope observations at different times showed that VE-C3 underwent two types of adhesion: (i) cell-to-cell interactions, preceding the cell adhesion to the n-alkane, and (ii) incorporation of nanodroplets of n-alkane into the hydrophilic CPS to form a more hydrophobic polysaccharide–n-alkane matrix surrounding the cell wall. The incorporation of n-alkanes as nanodroplets into the CPS of VE-C3 cells might ensure the partitioning of the bulk apolar phase between the aqueous medium and the outer cell membrane and thus sustain a continuous growth rate over a prolonged period.     

Nutrient speciation and hydrography in two anchialine caves in Croatia: tools to understand iodine speciation

Despite iodine being one of the most abundant of the minor elements in oxic seawater, the principal processes controlling its interconversion from iodate to iodide and vice versa, are still either elusive or largely unknown. The two major hypotheses for iodate reduction involve either phytoplankton growth in primary production, or bacteria during regeneration. An earlier study intended to exploit the unusual nature of anchialine environments revealed that iodide is oxidised to iodate in the bottom of such caves, whereas reduction of iodate occurs in the shallower parts of the water column. This investigation was made on the hypothesis that study of the nitrogen and phosphorus nutrient systems within the caves might offer a bridge between the iodine chemistry and the marine bacteria which are assumed to be the agent of change of the iodine in the caves. Accordingly, the hydrography, the nutrient chemistry, and some further iodine studies were made of two anchialine caves on the east coast of the Adriatic Sea in Croatia. Iodate and iodide were determined by differential pulse voltammetry and cathodic stripping square-wave voltammetry, respectively. Total iodine was determined indirectly, as iodate, after oxidation of reduced iodine species with UV irradiation and strong chemical oxidants. Nutrient concentrations were measured by spectrophotometry. Nutrient profiles within the well stratified water columns indicate a relatively short-lived surface source of nitrate and phosphate to the caves, with a more conventional, mid-water, nutrient regeneration system. The latter involves nitrite and ammonium at the bottom of the halocline, suggestive of both autotrophic and heterotrophic microbial activity. High iodate/low iodide deep water, and conservative behaviour of total inorganic iodine were confirmed in both systems. Iodate is reduced to iodide in the hypoxic region where nutrient regeneration occurs. The concentrations of organic iodine were surprisingly high in both systems, generally increasing toward the surface, where it comprised almost 80% of total iodine. As with alkalinity and silica, the results suggest that this refractive iodine component is liberated during dissolution of the surrounding karst rock. A major, natural flushing of one of the caves with fresh water was confirmed, showing that the cave systems offer the opportunity to re-start investigations periodically.     

PTCH1 gene polymorphisms in ovarian tumors: Potential protective role of c.3944T allele

In this study we investigated the types and role of different genetic changes of PTCH1 gene in three different types of ovarian tumors: carcinomas, fibromas and dermoids. LOH of the PTCH1 region was detected in 27.3% ovarian carcinoma samples, 18.18% ovarian fibroma samples and 55.56% ovarian dermoid samples. No point mutations were detected in any of the three types of ovarian tumors. SNP c.3944C>T showed significant differences between ovarian carcinoma and control samples with the minor T allele being significantly higher in controls compared to ovarian carcinomas. Interestingly, a new polymorphism c.−1184G>A was found only in tumor samples and further analyses should be performed in order to elucidate its potential role in ovarian tumors.