Labial Gland and Its Protein Patterns of Hydropsychid Caddisfly (Hydropsyche kozhantschikovi Martynov: Trichoptera)

ABSTRACT There is a pair of labial gland of hydropsychid caddisfly ( Hydropsyche kozhantschikovi Martynov) larva. It is in 'Z' formation in the body and the total length is about 20 mm. Hydropsychid caddishfly larvae that were raised under the lab conditions were able to form a nest-spining by connecting the small grains of sand provided. By repeatedly treating the extracted labial gland with methanol/D.W., the cell layer was removed. Accordingly, only the matrix within the labial gland that did not dissolve in water was obtained. The matrix inside the methanol/D.W. treated labial gland was dissolved with 5% acetic acid. Then the results of an acidic electrophoresis with a number of conditions indicated that 5% acetic acid/ 5 M urea/ 8% PAGE was the most effective. Moreover, the result of 2-D PAGE on the labial gland of these hydropsychid caddisfly larva, the number of proteins in the labial gland including the cell layer was about 350 and the number of proteins in the labial gland treated with methanol/D.W. was about 80, showing a substantially small number of proteins.     

Anti-neuroinflammatory effects of sesquiterpenoids isolated from Nardostachys jatamansi

Two new nardosinone-type sesquiterpenoids, namely kanshone J (1) and kanshone K (2) along with seven known terpenoids (3–9) were isolated from the rhizomes and roots of Nardostachys jatamansi DC (Valerianaceae). The structures of these compounds were determined mainly by analysis of 1D-, 2D-NMR and MS data. In addition, the absolute configuration of compound 1 was assigned by application of the modified Mosher’s method. In an initial assay to evaluate their anti-neuroinflammatory effects, compounds 1–5 and 9 exhibited dose-dependent inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in BV2 cells, with IC₅₀ values ranging from 2.43 to 46.54?μM. Particularly, desoxo-narchinol A (3) and narchinol B (4) significantly inhibited LPS-induced NO overproduction in BV2 cells with IC₅₀ values of 3.48?±?0.47 and 2.43?±?0.23?μM, respectively. Furthermore, compounds 3 and 4 exhibited anti-neuroinflammatory effects by inhibiting the production of pro-inflammatory mediators, including prostaglandin E₂ (PGE₂), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) proteins, and pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF)-α, in LPS-stimulated BV2 and primary microglial cells.     

Effect of medium milieu on sperm penetration and pronuclear formation of bovine oocytes matured in vitro

The purpose of the present study was to investigate the optimal concentration of osmolarity, calcium and bicarbonate for sperm penetration and formation of pronuclei (PN), and to investigate the time required for capacitation, penetration across the zona pellucida and formation of PN in bovine cumulus-free oocytes matured in vitro. Bovine follicular oocytes collected at slaughter were matured and fertilized in vitro. Bovine sperm penetrated the zona pellucida in medium containing 240 to 440 mOsm, whereas PN formation was observed in a narrow range of osmolarities, from 280 to 360 mOsm. Maximal penetration by spermatozoa and PN formation was obtained in the medium with 2.5 mM calcium. High rates of spermatozoa penetration were observed in the medium with 37 to 49 mM NaHCO3. However, PN were formed regardless of the concentration of NaHCO3. The times required for sperm capacitation and penetration through the zona pellucida were 260 and 50 min, respectively. The first development of PN was recorded at 120 min after sperm penetration. Therefore, our study suggests that fertilization ability of spermatozoa in vitro appears to be more stable in high concentrations of NaCI. Oocytes are more sensitive to osmotic stress than spermatozoa. Calcium is required for both sperm penetration and PN formation in cumulus-free oocytes, but bicarbonate may be needed mainly for the penetration of spermatozoa.