The Malassezia fungi are responsible for various human skin disorders including dandruff and seborrheic dermatitis. Of the Malassezia fungi, Malassezia globosa (M. globosa) is one of the most common in human scalp. The completed genome sequence of M. globosa contains four putative cytochrome P450 genes. To determine the roles of Malassezia P450 enzymes in the biosynthesis of ergosterol, we isolated MGL3996 gene from M. globosa chromosomal DNA by PCR. The MGL3996 gene encodes an enzyme of 616 amino acids, which shows strong similarity with known CYP52s of other species. MGL3996 gene was cloned and expressed in Pichia pastoris (P. pastoris) heterologous yeast expression system. Using the yeast microsomes expressing MGL3996 protein, a typical P450 CO-difference spectrum was shown with absorption maximum at 448 nm. SDS-PAGE analysis revealed a protein band of apparent molecular weight 69 kDa and Western blot with anti-histidine tag antibody showed that MGL3996 was successfully expressed in P. pastoris. Cloning and expression of a new P450 gene is an important step to study the P450 monooxygenase system of M. globosa and to understand the role of P450 enzymes in pathophysiology of dandruff.

BACKGROUND Femoral neck stress fractures (FNSFs) are commonly found in long-distance running athletes. For FNSFs, early diagnosis and proper treatment are important. The objective of this study was to report FNSFs that occurred after excessive exercise using trampoline in middle-aged women. CASE SUMMARY The patient was a 43-year-old woman who exercised jumping on a trampoline for 6 wk for 1-3 h a day to diet. Exercise includes repeated flexion-extension of the hip joint. The patient was admitted to the hospital due to sudden bilateral groin pain that occurred suddenly during a trampoline exercise. Hip magnetic resonance imaging (MRI) revealed bilateral FNSFs. After 2 wk of follow-up with conservative treatment, the pain slightly decreased. However, it did not disappear completely. It was determined that it was difficult to control symptoms only by conservation treatment. Thus, closed reduction and internal fixation using a cannulated screw were performed for the more painful left hip joint. After operation, the pain was improved. Walking using crutches was possible. Follow-up MRI showed that the right femoral head signal was decreased compared to the left femoral head signal. Therefore, nonsteroidal anti-inflammatory drug and conservative treatment were provided. CONCLUSION In middle-aged people, excessive trampoline exercise can repeat hip flexion and extension for a short period of time, leading to FNSFs.

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-Clusterin is a heterodimeric sulfated glycoprotein and plays a role in many different types of cancer as a cell survival factor and helps cancerous cells to evade stress-induced apoptosis. To investigate whether the regulation of clusterin expression is involved in the mechanism of anticancer agent, we studied the effect of tamoxifen on clusterin expression in human prostate cancer PC-3 cells. Treatment of PC-3 cells with tamoxifen reduced cellular proliferation. Western blot analyses showed that treatment with tamoxifen suppressed clusterin expression in a concentration-dependent manner. Transfection with clusterin siRNA plasmid showed that clusterin is required for PC-3 cell survival. We found that tamoxifen resulted in a rapid decrease in the phosphorylation of Akt on Ser473 leading to prevent kinase activity. Expression of myristoylated Akt prevented tamoxifen-mediated clusterin downregulation. Interestingly, MG132, a wellknown proteasome inhibitor also recovered clusterin expression suppressed by tamoxifen. These data indicate that clusterin expression may be regulated by activation of Akt and ubiquitin-proteasome pathway plays an important role in tamoxifen-mediated clusterin suppression.