Identification of a signal peptide for oryzacystatin-I.

A previously unidentified extension of an open reading frame from the genomic DNA of Japonica rice (Oryza sativa L.) encoding oryzacystatin-I (OC-I; access. M29259, protein ID AAA33912.1) has been identified as a 5' gene segment coding for the OC-I signal peptide. The signal peptide appears to direct a pre-protein (SPOC-I; Accession No. AF164378) to the endoplasmic reticulum, where it is processed into the mature form of OC-I. The start codon of SPOC-I begins 114 bp upstream from that previously published for OC-I. A putative proteolytic site. which may yield a mature OC-I approximately 12 residues larger than previously described, has been identified within SPOC-I between Ala-26 and Glu-27. The signal peptide sequence was amplified by polymerase chain reaction using genomic DNA from O. sativa seedlings and ligated to the 5' end of the truncated OC-I gene at the endogenous SalI site. Partially purified protein extracts from Escherichia coli expressing SPOC-I reacted with polyclonal antibodies raised against OC-I and revealed a protein of the expected molecular weight (15,355 Da). In-vitro translation of SPOC-I in the presence of microsomal membranes yielded a processed product approximately 2.7 kDa smaller than the pre-protein. Nicotiana tabacum L. cv. Xanthi plants independently transformed with the SPOC-I gene processed SPOC-I and accumulated the mature form of OC-I (approximately 12.6 kDa), which co-migrated with natural, mature OC-I extracted from rice seed when separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Outcomes of contemporary percutaneous nephrostolithotomy in morbidly obese patients.

PURPOSE: The surgical management of renal and ureteral calculi presents unique challenges in the obese patient. We reviewed our recent experience with percutaneous nephrostolithotomy in patients with a body mass index greater than 30. MATERIALS AND METHODS: We reviewed the records of 236 patients undergoing percutaneous nephrostolithotomy between August 1994 and March 1997 at 2 university and affiliated hospitals. Among this group 57 patients had a body mass index greater than 30 (mean plus or minus standard deviation 38.9 +/- 7.4). Renal anomalies were present in 21% of patients and more than half (56%) had co-morbidities in addition to obesity. Staghorn calculi were present in 19 patients (31.7%). In the remaining patients the mean cumulative stone size for single or multiple stones was 14.5 +/- 8.7 mm. Stone-free status after the procedure was determined by plain nephrotomograms or noncontrast computerized tomography. RESULTS: A total of 96 procedures were performed on 60 renal units (1.6 procedures per renal unit). Average operative time for the initial procedure was 181.2 +/- 91.4 minutes. Complications occurred in 8 patients (14%) and 5 (8.8%) received a blood transfusion. Average hospital stay was 4.9 +/- 3.1 days. The overall stone-free rate was 88.3%, and stone-free rate for staghorn calculi was 84.2% compared with 90.2% for nonstaghorn calculi. CONCLUSIONS: Percutaneous nephrostolithotomy in obese patients yields a stone-free rate that is comparable to that achieved in an unselected patient population. The complication rate, transfusion rate and hospital stay are also similar. Modifications to standard technique and instrumentation are sometimes necessary to perform percutaneous nephrostolithotomy in this group of patients.

Advanced transitional cell carcinoma of the upper urinary tract: patterns of failure, survival and impact of postoperative adjuvant radiotherapy.

PURPOSE: We review the outcome of patients with advanced stage III or IV transitional cell carcinoma of the upper urinary tract and the impact of postoperative radiotherapy. MATERIALS AND METHODS: We identified 74 patients who were treated surgically with curative intent for stage III (49) or IV (25) transitional cell carcinoma of the upper urinary tract. Median followup was 21 months (range 1 to 236) for all patients and 60 months (range 29 to 172) for those alive at last contact. A median dose of 40 Gy. adjuvant radiotherapy was delivered to the tumor bed and regional nodes of 15 patients (30%) with stage III and 13 (52%) with stage IV disease. RESULTS: The actuarial 5-year overall and disease specific survival for patients with stage III disease was 28 and 40%, respectively. Median disease specific survival was 37 months. Median overall and disease specific survival for patients with stage IV disease was 7 months. Isolated local recurrence was identified in 5 of 49 patients with stage III and only 1 of 25 with stage IV disease. The 5-year actuarial disease specific survival rate in patients with stage III disease whether or not they were treated with postoperative radiotherapy was 45 versus 40%, respectively. For patients with stage IV disease median survival was 7 and 9 months for those who were and those who were not treated with postoperative radiotherapy, respectively. CONCLUSIONS: Patients with stages III and IV transitional cell carcinoma of the upper urinary tract have a high risk of disease relapse and cancer mortality. The major clinical feature is distant failure with isolated local relapse uncommon following initial aggressive surgical therapy. There is no survival benefit with postoperative adjuvant radiotherapy. More effective systemic adjuvant therapy is necessary to improve the outcome of these patients.