Monthly Archives: March 2015

USP8, a Regulator of Endosomal Sorting, Is Involved in Mouse Acrosome Biogenesis Through Interaction with the Spermatid ESCRT-0 Complex and Microtubules: Recombinant Proteins

Production and Purification of Recombinant Proteins The following constructs for expression of glutathione S-transferase (GST) fusion proteins, namely, were used: GST-USP8542-1080 (kindly provided by Dr. E. Martegani, University of Milano-Bicocca, Italy), GST-USP81_438, and GST-USP81-m (kindly provided by Dr. S. Urbe, School of Biomedical Sciences, Liverpool, U.K.), and GST-USP8542–660. GST-USP8542_1Q80 fusion construct contains the second SH3-binding […]

USP8, a Regulator of Endosomal Sorting, Is Involved in Mouse Acrosome Biogenesis Through Interaction with the Spermatid ESCRT-0 Complex and Microtubules: Immunoblotting Analysis

Subcellular Protein Fractionation and Immunoblotting Analysis One-percent Triton X-100-soluble protein lysates and hypotonic protein extracts from freshly isolated spermatogenic cells were obtained as described. To separate cytosolic and membrane particulate fractions, the hypotonic extract from postnuclear supernatant was spun at 100 000 X g for 1 h at 4°C. The resulting supernatant is the cytosolic […]

USP8, a Regulator of Endosomal Sorting, Is Involved in Mouse Acrosome Biogenesis Through Interaction with the Spermatid ESCRT-0 Complex and Microtubules: MATERIALS AND METHODS

Animals Testes were collected from 3- to 4-mo-old CD1 male mice (Charles River, Calco, Italy). Handling of mice and experimental procedures were reviewed and approved by the Ethical Committee of the University of Milano and were performed in accordance with the Guidelines for the Care and Use of Laboratory Animals promulgated by the Italian Minister […]

USP8, a Regulator of Endosomal Sorting, Is Involved in Mouse Acrosome Biogenesis Through Interaction with the Spermatid ESCRT-0 Complex and Microtubules

The deubiquitinating enzyme USP8 (ubiquitin-specific peptidase 8; previously named UBPy, ubiquitin-specific processing protease-y) was originally identified as a putative protein encoded by a cDNA expressed in human myeloblasts (hUBPy) and then found, through independent yeast two-hybrid screenings of mouse libraries, as a protein (mUBPy) that interacts with STAM2 (also known as Hbp) and the Ras-guanine […]

Tracheobronchial Amyloidosis: Conclusion

Kurrus et al reported one of the first intentional uses of external beam radiation therapy for TBA. The patient was initially observed until the development of life-threatening, progressive obstruction. External beam radiation therapy was then delivered to the distal trachea and right mainstem bronchus using a dose of 20 Gy in 10 fractions with improvement. […]

Tracheobronchial Amyloidosis: Pulmonary

Management has primarily consisted of debulking procedures and laser ablation. Unfortunately, such methods may be ineffective and repeated procedures may be required. Capizzi et al reported 17 patients with TBA who were treated with laser or forceps resection, systemic therapy, radiation, or observation. Of 13 patients with follow-up data, 6 patients were alive with recurrence, […]