Essential oil variation among and within six Achillea species transferred from different ecological regions in Iran to the field conditions

The compositions of essential oils of 19 accessions belonging to six different Achillea species, transferred from the natural habitats in 10 provinces of Iran to the field conditions, were assessed. The relationship between the leaf areas of selected accessions with their essential oil content was also investigated. Essential oil yield of dried plants obtained by hydro-distillation ranged from 0.1 to 2.7% in leaves. Results indicated a significant variation in oil composition among and within species. Total of 94 compounds were identified in 19 accessions belonging to the six species of A. millefolium, A. filipendulina, A. tenuifolia, A. santolina, A. biebersteinii and A. eriophora. The major constituents of the leaves in the tested genotypes were determined as germacrene-D, bicyclogermacrene, camphor, borneol, 1,8-cineole, spathulenol and bornyl acetate. According to the major compounds, four chemotypes were defined as: (I) spathulenol (1.64–34.31%) + camphor (0.2–15.61%) (7 accessions); (II₁) germacrene-D (18.78–23.93%) + borneol (7.93–8.26%) + bornyl acetate (11.56–14.66%) (5 accessions); (II₂) germacrene-D (13.28–36.28%) + bicyclogermacrene (5.93–8.4%) + 1,8-cineole (15.26–19.41%) + camphor (14.95–23.32%) (2 accessions); (III) borneol + camphor (52.04–63.27) (2 accessions); (IV) germacrene-D (45.86–69.64%) (3 accessions). The relationships of chemotypes with soil type and climatic conditions of collected regions were assessed, as probable reasons of high variations in essential oil components, and discussed.     

Comparative Study of Immunological and Structural Properties of Two Recombinant Vaccine Candidates against Botulinum Neurotoxin Type E

Background

Recently, botulinum neurotoxin (BoNT)-derived recombinant proteins have been suggested as potential botulism vaccines. Here, with concentrating on BoNT type E (BoNT/E), we studied two of these binding domain-based recombinant proteins: a multivalent chimer protein, which is composed of BoNT serotypes A, B and E binding subdomains, and a monovalent recombinant protein, which contains 93 amino acid residues from recombinant C-terminal heavy chain of BoNT/E (rBoNT/E-HCC). Both proteins have an identical region (48 aa) that contains one of the most important BoNT/E epitopes (YLTHMRD sequence).

Methods

The recombinant protein efficiency in antibody production, their structural differences, and their BoNT/E-epitope location were compared by using ELISA, circular dichroism, computational modeling, and hydrophobicity predictions.

Results

Immunological studies indicated that the antibody yield against rBoNT/E-HCC was higher than chimer protein. Cross ELISA confirmed that the antibodies against the chimer protein recognized rBoNT/E-HCC more efficiently. However, both antibody groups (anti-chimer and anti-rBoNT/E-HCC antibodies) were able to recognize other proteins. Structural studies with circular dichroism showed that chimer proteins have slightly more secondary structures than rBoNT/E-HCC.

Conclusion

The immunological results suggested that the above-mentioned identical region in rBoNT/E-HCC is more exposed. Circular dichroism, computational protein modeling and hydrophobicity predictions indicated a more exposed location for the identical region in rBoNT/E-HCC than the chimer protein, which is strongly in agreement with immunological results. Iran. Biomed. Key Words: Botulinum neurotoxin type E (BoNT/E), Cross ELISA, circular dichroism, Computational modeling, recombinant vaccine-candidates     

Araguspongine C Induces Autophagic Death in Breast Cancer Cells through Suppression of c-Met and HER2 Receptor Tyrosine Kinase Signaling

Receptor tyrosine kinases are key regulators of cellular growth and proliferation. Dysregulations of receptor tyrosine kinases in cancer cells may promote tumorigenesis by multiple mechanisms including enhanced cell survival and inhibition of cell death. Araguspongines represent a group of macrocyclic oxaquinolizidine alkaloids isolated from the marine sponge Xestospongia species. This study evaluated the anticancer activity of the known oxaquinolizidine alkaloids araguspongines A, C, K and L, and xestospongin B against breast cancer cells. Araguspongine C inhibited the proliferation of multiple breast cancer cell lines in vitro in a dose-dependent manner. Interestingly, araguspongine C-induced autophagic cell death in HER2-overexpressing BT-474 breast cancer cells was characterized by vacuole formation and upregulation of autophagy markers including LC3A/B, Atg3, Atg7, and Atg16L. Araguspongine C-induced autophagy was associated with suppression of c-Met and HER2 receptor tyrosine kinase activation. Further in-silico docking studies and cell-free Z-LYTE assays indicated the potential of direct interaction between araguspongine C and the receptor tyrosine kinases c-Met and HER2 at their kinase domains. Remarkably, araguspongine C treatment resulted in the suppression of PI3K/Akt/mTOR signaling cascade in breast cancer cells undergoing autophagy. Induction of autophagic death in BT-474 cells was also associated with decreased levels of inositol 1,4,5-trisphosphate receptor upon treatment with effective concentration of araguspongine C. In conclusion, results of this study are the first to reveal the potential of araguspongine C as an inhibitor to receptor tyrosine kinases resulting in the induction of autophagic cell death in breast cancer cells.     

Relationship Between Degree of Starch Gelatinization and Quality Attributes of Parboiled Rice During Steaming

Paddy rice samples were parboiled by soaking at 65 °C for 180 min and steaming at 96 °C for 2–10 min, and then dried to achieve the final moisture content of 11% ± 1%. The degree of starch gelatinization(DSG) and several quality attributes(head rice yield(HRY), color value and hardness) of parboiled rice were measured. Results showed that DSG(46.8%–77.9%), color value(18.08–19.04) and hardness(118.6–219.2 N) all increased following steaming. In contrast, the HRY increased(64.8%–67.1%) for steaming times between 2–4 min but decreased(67.1%–65.0%) for steaming times between 4–10 min. Linear relations between DSG and color value(R~2 = 0.87), and DSG and hardness(R~2 = 0.88) were observed. The suitable DSG of parboiled rice leading to the highest HRY was found to be 62.5%, obtained following 4 min of steaming.     

Evaluation of Plasticizing and Antioxidant Properties of Silver Carp Protein Hydrolysates in Fish Gelatin Film

Physico-mechanical and antioxidant properties of glycerol plasticized fish gelatin films were investigated at different levels of fish protein hydrolysates (FPH) from silver carp muscle (5, 10, 15, and 20 g FPH/100 g gelatin). The films supplemented with FPH, especially at higher concentration showed significant increase in 2, 2-diphenyl-1-picrylhydrazyl (DPPH), 2, 20-azinobis-(3-ethylbenzothiazoline-6-sulfnic acid) (ABTS) radical scavenging, and ferric reducing antioxidant power (FRAP), elongation at break, difference of color, water vapor permeability and opacity, while tensile strength, elastic modulus, L*, b*, and contact angle decreased markedly (p < 0.05). However, no significant differences were observed for a* and solubility of all samples (p > 0.05). Films prepared using 20% FPH had lower glass transition temperature and more homogeneous structure, compared with control film. Also, Fourier transform infrared (FTIR) spectroscopy confirmed the increase of free groups of hydrolysates and lower interaction between chains of film incorporated with the highest content of FPH. Thus, these results indicate that FPH enhanced the antioxidant activities and affected some characteristics due to less interaction between gelatin and FPH.     

Pullularins E and F, two new peptides from the endophytic fungus Bionectria ochroleuca isolated from the mangrove plant Sonneratia caseolaris

Chemical investigation of the EtOAc extract of the endophytic fungus Bionectria ochroleuca, isolated from the inner leaf tissues of the plant Sonneratia caseolaris (Sonneratiaceae) from Hainan island (China), yielded two new peptides, pullularins E and F (1 and 2) together with three known compounds (3–5). The structures of the new compounds were unambiguously determined on the basis of one- and two-dimensional NMR spectroscopy as well as by high-resolution mass spectrometry. The absolute configurations of amino acids were determined by HPLC analysis of acid hydrolysates using Marfey’s method. The isolated compounds exhibited pronounced to moderate cytotoxic activity against the mouse lymphoma cells (L5178Y) with EC₅₀ values ranging between 0.1 and 6.7 µg/mL.     

The correlation between the endometrial integrins and osteopontin expression with pinopodes development in ovariectomized mice in response to exogenous steroids hormones

Background: The ovariectomized animals are good models to evaluate the effect of different steroid hormone treatments on implantation events and the pattern of integrin expression. Therefore, this study was performed to compare the expression of integrins and osteopontin (OPN) in correlation with pinopode development in ovariectomized mice endometrium which was subjected to steroid hormones. Methods: Ovariectomized mice were subjected to estrogen, progesterone and estrogen-progesterone hormones. Their uterine horns were evaluated for integrin expression by immunohistochemistry and real-time RT-PCR and for pinopode development by transmission and scanning electron microscopic studies. Results: No immunostaining for integrin and OPN molecules were detected in the endometrium of non-ovariectomized mice except in metestrus phase. The α4 and β1 integrin genes were expressed in all phases of estrous cycle. In ovariectomized mice, no reaction was detected in the endometrium of control, sham and estrogen-treated groups, but in both progesterone-treated groups, all examined genes were expressed. There was not any correlation between pinopodes and integrin expression in ovariectomized mice. Conclusion: The progesterone is more effective on endometrial integrin expression than estrogen and differences in the expression pattern of integrins reflect their important and different roles in embryo implantation. The pinopodes may have minor effect in mice implantation or have some delay in their expressions in ovariectomized mice which were subjected to exogenous hormones. Key Words: Endometrium, Estrogen, Integrin, Ovariectomized mice, Osteopontin (OPN)     

Determining and mapping some soil physico-chemical properties using geostatistical and GIS techniques in the Naqade region,Iran

The distribution variability of soil electrical conductivity (EC), pH, clay, sand, CaCO₃, organic carbon (OC) and available potassium (K) in the Naqade region was investigated using a geostatistical method and Geographical Information System (GIS) technique. Two hundred and eighty-two topsoil (0–30 cm) samples were randomly collected and analyzed. pH and clay followed a normal distribution, whereas sand EC, CaCO₃, OC and K were log-transformed. The highest variation was observed for soil EC, and the lowest for soil pH. In the variography analysis, spherical, exponential and gaussian models were best fit on experimental semivariograms. The minimum effective spatial autocorrelation was 1500 m for OC and the maximum effective spatial autocorrelation was 4000 m for sand and K. Strong spatial correlations were noted with sand and CaCO₃ (<25%), whereas values were moderate for clay, EC, OC and K (25–75%). Ordinary kriging was utilized for the interpolation of estimated variable determinations in unsampled sites. It was found that soil properties in this study area were strongly influenced by both environmental and natural factors. The results can be used as a source of information for the development and implementation of any further land management and soil and water conservation plans.     

Molecular identification of 16SrII-D subgroup phytoplasmas associated with chickpea and faba bean in Sudan

In January 2011, symptomatic chickpea and faba bean plants were observed in fields located in the Gezira state (Sudan). Faba bean plants showed yellowing and stunting, whereas chickpea plants presented yellowing, reddening and little leaves. The disease etiology was investigated using nested polymerase chain reaction (PCR) with phytoplasma-specific primers which amplify a fragment of the 16S rRNA gene. Sequencing and restriction fragment length polymorphism (RFLP) analyses revealed that the tested phytoplasmas belonged to the group 16SrII. Phylogenetic analyses of the 16S rRNA gene of the obtained sequences indicated that the chickpea and faba bean phytoplasmas from Sudan were more closely related to the phytoplasmas subgroup 16SrII-D. To our knowledge, this is the first report of phytoplasmas from the group 16SrII-D infecting chickpea in Sudan, and faba bean worldwide.