Pellino, a highly conserved E3 ubiquitin ligase, is known to mediate ubiquitination of phosphorylated Interleukin-1 receptor-related kinase (IRAK) homologs in Toll signaling pathway. To understand the immunological function of TmPellino, we screened the knockdown efficiency of TmPellino by injecting TmPellino-specific dsRNA into T. molitor larvae. Subsequently, we investigated the larval mortality and the tissue-specific expression patterns of antimicrobial peptide (AMP) genes against microbial challenges. Interestingly, the results indicate that the expression of many AMP genes was upregulated in the Malpighian tubules of TmPellino-silenced T. molitor larvae. This study may provide basic information to understand how Tmpellino regulates AMPs production in T. molitor.

Earthworms, especially Eisenia andrei, are cultivated for a variety of purposes including waste disposal and compost production. In this study, bacteria from earthworm gut were cultured and the dominant species identified. Subsequently, we isolated bacteriophages able to lyse the isolated gut bacteria. Two dominant genera of gut bacteria, Aeromonas sp. and Citrobacter sp. were identified by using MALDI-TOF MS analysis and a library was constructed to find lytic phages. Phage EF1 showed lytic activity for C. freundii and two Citrobacter isolates, C. braakii and C. murliniae. These 3 species have similar sensitivities to EF1. Several aspects of the life cycle of EF1 were investigated by using C. freundii under optimal growth conditions. EF1 infects C. freundii with a moderate latent period, approximately 25 min, and a large burst size averaging 5 × 109 per infected cell. Moreover, the antimicrobial activity of EF1 was well maintained under diverse conditions including a broad temperature range of 40°C to 50°C and a wide pH range of 4 to 11. In conclusion, the results indicate that earthworm casting contain a wide range of bacteria species, for which there are various corresponding bacteriophages.

Background : The effective components of Omija(Schisandra chinensis Bailllon) are lignans (schizandrins and gomisins), and this components were contented mostly in seed part on Omija, which have various physiological functionalities such as anti-cancer, anti-inflammatory, and antioxidant activities. Methods and Results : This study was carried out to determine effective condition(CO2, CO2+ethanol) on extraction using supercritical carbon dioxide extraction (SFE) system and to find interrelation on effective components and antioxidant activity of extracts and residues obtained after extraction. Effective components were analysed lignans and phenolic compounds and antioxidant activirty was determined for DPPH radical scavenging ability on methanol extracts of SFE-extract and SFE-residue. On SFE with ethanol, SFE extract was separated two phase, upper(water phase) and lower(oil phase). SFE-extract showed the highest total lignans content(61.36 mg/g, 72.14 mg/g on lower, 50.58 mg/g on upper) and the lowest total phenolic compounds(6.52 mg/100g) and SFE-residue showed the lowest total lignans content(1.45 mg/g) and the highest total phenolic compounds(16.23 mg/100g) by extracted on CO2+ethanol treatment. SFE-residue methanol extract showed the highest DPPH radical scavenging abilities and SFE-extract upper showed the lowest. Conclusion : Thus, this results showed SFE-extract showed the highest total lignans content, but SFE-residue showed the highest DPPH radical scavenging ability although the lowest total lignans content.

After vasectomy (VAX), the flux and composition of the epididymal fluid are modified, leading to sequelae to the epididymis. In an effort to understand molecular pathophysiology of the epididymis following VAX, we investigated the changes of gene expression and sperm functions in the epididymis of vasectomized mice. After VAX, the epithelial cell height was significantly decreased in cauda epididymis, resembling the those of vas deference. This suggests that VAX evokes alteration of segmental characteristics of epididymal epithelium. Of note, these was an increase in luminal diameter in corpus and cauda epididymis, indicating the alteration of fluid homeostasis in epididymal lumen and protein synthesis and secretion. Also, the formation of sperm granuloma and infiltration of inflammatory cells were noted in lumen of epididymis at 8 weeks postvasectomy, indicating the activation of immune response in epididymis. The serum TNF-α levels and epididymal TNF-α and IL-1β mRNA levels were significantly increased in VAX mice. Microarray analysis demonstrated that claudin (CLDN) 10 and cystic fibrosis transmembrane conductance regulator (CFTR) were downregulated after vasectomy. In contrast, angiotensin II receptor type 2 (Agtr2) was up-regulated after vasectomy. Taking into account the functional importance of angiotensin system in epididymal epithelia and muscle tissue, VAX may alter the secretory function of epithelia and sperm transport via alteration in angiotensin system. Importantly, the IgG type of anti-sperm antibody (ASA) were markedly increased in the blood of vasectomized mice. Together this indicates that VAX provokes local inflammation in epididymis as well as systemic inflammation, which in turn change the blood epididymal barrier, an important element for epididymal immunological privilege. In addition, the number of sperm in cauda epididymis was increased but the sperm motility was decreased after vasectomy. The spontaneous acrosome reaction was increased by vasectomy after capacitation. This suggests that VAX affects sperm functions as well as sperm maturation. In conclusion, bilateral vasectomy lead to local immune response of epididymis, causing immunologic infertility in men.

The Egr family of zinc finger transcription factors is rapidly induced by various mitogens and regulates cell growth, differentiation, and apoptosis. While it is clear that loss of Egr1 leads to anovulatory infertility due to LHβ deficiency in female mice, molecular function of Egr1 in male reproduction has not been clearly investigated. Here, we demonstrate that Egr1 acts as an intrinsic transcription factor in Leydig cells to regulate their proliferation and steroidogenesis in the testis as well as an extrinsic factor for male reproduction via LHβ transcription in the pituitary. Egr1 is predominantly expressed in spermatogonia and Leydig cells in immature testes and later detected in some of these cell types in mature testes. The fertility potential of Egr1(-/-) male mice is relatively deteriorated even at 2 month-old age and aggravated with aging. The incidence of abnormalities of seminiferous tubules such as Sertoli cell only was dramatically increased with aging. The number and mean size of Leydig cells were significantly reduced in Egr1(-/-) testes. The impairment of Leydig cells is consistent with significant reduction in levels of testosterone and expression of factors critical for steroidogenesis such as StAR in Egr1(-/-) testes. Exogenous administration of hCG rapidly and transiently induced Egr1 expression in Leydig cells culture in vitro. hCG could reinstate reduced mean size of Leydig cells but not reduced number of Leydig cells and aberrantly low StAR expression, suggesting that Egr1 has critical functions for Leydig cell proliferation and their steroidgenesis. In addition, daily sperm production and in vitro fertilization (IVF) competence were significantly reduced, and apoptosis was facilitated in these mice. Furthermore, hCG administration to compensate for relatively low LH levels in Egr1(-/-) males could not restore the compromised reproductive phenotypes such as IVF competence and apoptosis in these mice. Interestingly, expression of Egr2, a member of Egr family, is significantly elevated in Egr1(-/-) Leydig cells suggesting that genetic compensation of Egr2 may alleviate phenotypic aberration of Egr1(-/-) male testes. Collectively, these results suggest that Egr1 act as an intrinsic transcription factor required for proliferation and steroidogenesis of Leydig cells to govern spermatogenesis in the testis.