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   Background & objectives: Calprotectin, S100A8 and S100A9 are involved in important processes including cell signaling and regulation of inflammatory responses. In this study, recombinant expression, purification and structural characterization of S100A8 and S100A9 were accomplished.

  Methods : In this experimental study, pET15b was used as vector of human S100A8 and S100A9 coding sequences, hosted by E.coli BL21 (DE3). Gene expression and purification attempts were evaluated using SDS-PAGE. Protein purification was accomplished using Ni-NTA resin based on its affinity for His-tag present on recombinant proteins. Tertiary structure of proteins were evaluated using spectrofluorimetry.

  Results : The subunits were over expressed 3-4 hours following induction at 37 °C. S100A9 was expressed mainly as inclusion body while S100A8 was found to be expressed mainly as a soluble protein. Purification of S100A8 and S100A9 was achieved at 100 mM imidazole. Spectroscopic studies showed that the amino acid tryptophan is in the internal structures and is less exposed to the aqueous environment.

  Conclusion : In this study, a recombinant S100A8 and S100A9 subunits were expressed and purified and also their structures were confirmed.

 

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Background & objectives: Artemisia absinthium (known as wormwood) is used as an antihelminthical, antimalarial, antiseptic and  anti-inflammatory agent, and is used for treatment of gastric pain in the traditional medicine. In the present study, we evaluated the effects of hydroalcoholic extract of A. absinthium on the ovary tissues and pituitary_gonadal axis of the adult female NMRI mice.
Methods: In this experimental study, intraperitoneally (IP) injections of hydroalcoholic extract of A. absinthium, were performed over 30 days on 50 mice with 50 (first group), 100 (second group), and 150 mg/kg.bw (third group) doses.  The sham group was received distilled water and control group received no injection. After 30 days of injections, the animals were dissected, and blood samples were collected by heart aspiration.  The levels of LH, FSH, estrogen and progesterone of serum were measured by ELISA method. Seven µm sections of ovary were prepared by a microtome and stained by H&E method. Data were analyzed by One-Way ANOVA and Tukey post- hoc test. The Significance level was set at p<0.05.       
Results: Our findings indicated a significant reduction (p<0.05) in the body weight in all experimental groups compared with sham and control groups. In parameters such as: large and small diameters of the ovary , number of primary, secondary, growing, graafian follicles, and corpus luteum, a significant decrease was observed in 100 and 150 mg/kg doses (p<0.05). In all experimental groups, no significant changes were observed in estradiol and progesterone levels. However, FSH and LH showed a significant decrease in 100 and 150 mg/kg doses (p<0.05).
Conclusion: Therefore, it can be concluded that Artemisia absinthium hydroalcohlic extract in high doses has damaging effects on the process of oogenesis, which may be due to α and β Thujoun, Alkaloid and Saponin constituents in this plant.