Effects of monomethoxypolyethylene glycol-chitosan nanoparticle-mediated dual silencing of livin and survivin genes in prostate cancer PC-3M cells.

Monomethoxypolyethylene glycol-chitosan (mPEG-CS) nanoparticles were used as interfering RNA carriers to transfect human prostate cancer PC-3M cells to evaluate the effects of livin and survivin gene silencing on the proliferation and apoptosis. mPEG-CS nanoparticles with sizes of approximately 60 nm were first synthesized by ionic crosslinking. Through electrostatic adsorption, mPEG-CS-livin short hairpin RNA (shRNA), mPEG-CS-survivin shRNA, and mPEG-CS-(livin shRNA + survivin shRNA) nanoparticles were then prepared to transfect PC-3M cells. The mRNA and protein expression levels of livin and survivin were measured by reverse transcription-PCR and western blotting, respectively. The inhibitory effects of down-regulated livin and survivin gene expression on the cell proliferation were evaluated by MTT assay. Cell apoptosis was assessed visually using Hoechst staining. Livin and survivin expression levels in all shRNA interference groups were effectively down-regulated at both the mRNA and protein levels. Dual silencing of livin and survivin genes markedly inhibited cell proliferation and facilitated apoptosis, with better outcomes than those of individual shRNA treatments. mPEG-CS nanoparticle-mediated dual shRNA interference of livin and survivin genes significantly reduced the expression levels in PC-3M cells, inhibited proliferation, and promoted apoptosis. As these effects were superior to single interference, this method may have synergistic effects.

The use of mannitol in partial and live donor nephrectomy: an international survey.

PURPOSE: Animal studies have shown the potential benefits of mannitol as renoprotective during warm ischemia; it may have antioxidant and anti-inflammatory properties and is sometimes used during partial nephrectomy (PN) and live donor nephrectomy (LDN). Despite this, a prospective study on mannitol has never been performed. The aim of this study is to document patterns of mannitol use during PN and LDN. MATERIALS AND METHODS: A survey on the use of mannitol during PN and LDN was sent to 92 high surgical volume urological centers. Questions included use of mannitol, indications for use, physician responsible for administration, dosage, timing and other renoprotective measures. RESULTS: Mannitol was used in 78 and 64 % of centers performing PN and LDN, respectively. The indication for use was as antioxidant (21 %), as diuretic (5 %) and as a combination of the two (74 %). For PN, the most common dosages were 12.5 g (30 %) and 25 g (49 %). For LDN, the most common doses were 12.5 g (36.3 %) and 25 g (63.7 %). Overall, 83 % of centers utilized mannitol, and two (percent or centers??) utilized furosemide for renoprotection. CONCLUSIONS: A large majority of high-volume centers performing PN and LDN use mannitol for renoprotection. Since there are no data proving its value nor standardized indication and usage, this survey may provide information for a randomized prospective study.

Making sense of postoperative CT imaging following laparoscopic partial nephrectomy.

The increasing popularity of laparoscopic partial nephrectomy (LPN) necessitates radiologists to become familiar with the operative techniques as well as normal and abnormal postoperative findings. Due to the varying presentation of abnormal changes following LPN and their similarities with other disease entities, radiologists should be cognizant of common pitfalls to avoid inadvertent misdiagnosis. A few common pitfalls discussed in this paper are the identification of laparoscopic port placement issues, recognizing a myriad of post-surgical materials, differentiating haemostatic materials from postoperative abscess and infection, non-absorbable suture material mimicking rim calcifications, as well as hints for differentiating exuberant granulation tissue from tumour recurrence.

Asymptomatic bacteriuria in candidates for active treatment of renal stones: results from an international multicentric study on more than 2600 patients.

The occurrence of asymptomatic bacteriuria concomitant to urolithiasis is an issue for patients undergoing renal stone treatment. Disposing of a preoperative urine culture is essential to reduce the risk of septic events. The endpoint of the study is to report which characteristics of candidates for renal stone treatment are frequently associated with positive urine culture. 2605 patients were retrospectively enrolled from 14 centers; inclusion criteria were age > 18 and presence of a single renal stone 1-2 cm in size. The variables collected included age, gender, previous renal surgery, comorbidities, skin-to-stone distance, stone size, location, density, presence of hydronephrosis. After a descriptive analysis, the association between continuous and categorical variables and the presence of positive urine culture was assessed using a logistic regression model. Overall, 240/2605 patients (9%) had preoperative bacteriuria. Positive urine culture was more frequent in females, patients with previous renal interventions, chronic kidney disease, congenital anomalies, larger stones, increased density. Multivariate analysis demonstrated that previous renal interventions (OR 2.6; 95% CI 1.9-3.4; p < 0.001), renal-related comorbidities (OR 1.31; 95% CI 1.19-1.4; p < 0.001), higher stone size (OR 1.06; 95% CI 1.02-1.1; p = 0.01) and density (OR 1.00; 95% CI 1.0-1.00; p = 0.02) were associated with bacteriuria; male gender and lower caliceal location were inversely related to it. Beyond expected risk factors, such as female gender, other parameters are seemingly favoring the presence of positive urine culture. The awareness of variables associated with bacteriuria allows to assess which individuals are at increased risk of presenting bacteriuria and reduce the rate of septic complications.

Precise characterization of urinary tract innervation using three-dimensional reconstruction: A contemporary review. / Caracterización precisa de la inervación del tracto urinario mediante reconstrucción tridimensional: una revisión contemporánea.

A precise understanding of the autonomic innervation of the urinary tract is crucial to successful management of urologic disease given the important role that neurophysiology plays in genitourinary pathology. Recent studies using a combination of contemporary histopathology and imaging technologies have furthered our understanding of the spatial nerve distribution in the kidneys, ureters, and bladder. The findings of these recent studies may have important clinical applications in expanding our knowledge of the etiology and treatment of disease processes affecting the urinary tract. In this narrative review, our goal is to provide an overview of the autonomic innervation of the urinary tract. Specifically, we aim to provide a three-dimensional gender-specific description of renal, ureteral and vesical innervation. We also highlight some possible opportunities for clinical and investigational application of this new knowledge.

Study of petrolatum structure: Explaining its variable rheological behavior.

The rheological properties of petrolatum are dependent on both temperature and thermal history. How this thermal dependency can be explained is unclear. In the past it has been suggested that the structure of petrolatum consists of a three-dimensional crystalline network. This has been established using old microscopic techniques only. Therefore a study on the microstructure of petrolatum was conducted using rheometry, DSC, pulsed NMR, polarized light microscopy and synchrotron X-ray. The combination of these techniques show that petrolatum is composed of 21% solid material at room temperature. This consists of partly crystalline lamellar sheets which are packed in stacks. The occurrence of these lamellar sheets is temperature dependent and the number of lamellar stacks is dependent on thermal history. It was shown that rheological differences in petrolatum can be explained by the number of lamellar stacks present, where more lamellar stacks result in more rigid petrolatum.

Combined continuous infusion etoposide with high-dose cyclophosphamide for refractory neuroblastoma: a phase II study from the Société Française d'Oncologie Pédiatrique.

PURPOSE: Patients older than 1 year with stage IV neuroblastoma who fail to achieve complete remission (CRem) have a particularly poor long-term prognosis. In an attempt to improve the outcome of these refractory patients, we tested a new drug combination. PATIENTS AND METHODS: Twenty-nine children with advanced neuroblastoma (27 stage IV and two stage III) were entered onto this phase II study. All were refractory to conventional chemotherapy and had measurable disease at the time of the trial. The regimen was a combination of high-dose cyclophosphamide (2 g/m2/d) on days 2, 3, and 4, and etoposide (VP16; 50 mg/m2/d) by continuous intravenous (IV) infusion on days 1 to 5. A pharmacokinetic study of VP16 was conducted in eight patients to determine whether the goal of persistent plasma levels between 1 and 5 micrograms/mL was achieved. RESULTS: Patients received a median of two courses, for a total of 58 courses. The median interval between each course was 32 days. In the 28 assessable patients, the overall response rate was 43%, with one CRem and 11 partial remissions (PRems). No life-threatening complication was observed in these heavily pretreated patients. The median duration of neutropenia (< 5 x 10(9)/L) was 14 days, and that of thrombocytopenia (< 50 x 10(9)/L) was 11 days. The overall incidence of sepsis was 27%. Gastrointestinal toxicity was frequent, but mild. Electrolyte disturbance with antidiuretic hormone (ADH)-like syndrome occurred in eight courses, but resolved rapidly. Grade > or = 2 hemorrhagic cystitis was observed in three courses. No cardiac toxicity was observed. There were no treatment-related deaths. Pharmacokinetic analysis showed that mean steady-state plasma levels (Css) of VP16 were greater than 1 microgram/mL during all the courses. CONCLUSION: This new drug combination appears to be effective in advanced neuroblastoma. Its toxicity remains manageable, with no life-threatening complications. Further evaluation in patients with less-advanced disease is warranted.

Listing of common HLA alleles and haplotypes based on the study of 356 families residing in the Paris, France, area: implications for unrelated hematopoietic stem cell donor selection.

In this study we have identified frequent human leukocyte antigen (HLA)-A, -B, -C,-DRB1, and -DQB1 alleles, frequent HLA-B/C, HLA-DRB1/DQB1 two-allele associations, and the most common HLA-A/B/C/DRB1/DQB1 five-locus haplotypes in a population residing in the Paris, France, area. The study was carried out in 356 families of children awaiting hematopoietic stem-cell transplantation (HSCT), with the selection criterion that haplotypes could be assigned with certainty to both the patient and at least one parent. Parental haplotypes were HLA-A, -B serologically typed, and HLA-C, -DRB1, -DQB1 broadly typed by polymerase chain reaction-sequence-specific oligonucleotide probe. The alleles of the most frequent haplotypes were subsequently defined at a high-resolution level by polymerase chain reaction-sequence-specific primer. The results on the distribution of common alleles and common allele associations demonstrated similarities with the previously published data in Caucasian populations, as expected from the geographic origin of the studied population. More importantly, this study provides the largest listing of common B/C and DRB1/DQB1 associations and of common five-allele haplotypes defined with certainty in a Caucasian population to date. These results can be used to help estimate the likelihood of finding a suitable donor in unrelated HSCT and to delineate search strategies for potential donors.

Evaluation of surgical performance with standard rigid and flexible-tip laparoscopes.

BACKGROUND: Flexible-tip laparoscopes have recently been introduced into clinical practice, with the goal of improving surgeon performance during complex laparoscopic procedures. We used objective and subjective performance parameters to compare standard rigid 0 degrees and 30 degrees lens laparoscopes two flexible-tip laparoscopes in an in vitro model. METHODS: Twenty-nine subjects with varied levels of surgical experience performed complex laparoscopic tasks in three different models simulating (a) prostate dissection from the rectum, (b) cystic duct clipping, and (c) distal posterior rectum dissection. Each task was performed using two Storz rigid laparoscopes (0 degrees and 30 degrees) and two flexible-tip laparoscopes, the Olympus LTF-V3 and the Fujinon EL2-TF310. The sequence of application of the two flexible-tip laparoscopes was randomized. In each case, an experienced laparoscopic camera driver controlled the field of vision. Time to complete each task, operative precision, and subjective surgeon rating scores were compared. Statistical analysis was performed with analysis of variance (ANOVA) and a two-sided fisher's exact test. RESULTS: In all three models, the flexible laparoscopes offered no advantage in terms of procedure time, surgical precision, or subjective surgeon rating score when compared with the 30 degrees lens rigid laparoscope. The 30 degrees rigid lens laparoscope and the two flexible-tip laparoscopes were superior to the 0 degrees lens rigid laparoscope for all parameters evaluated, with the exception of subjective rating in the cystic duct model and procedure time in the colorectal model. CONCLUSION: In this in vitro experimental model, the flexible-tip laparoscopes found to have no advantage over the standard rigid 30 degrees lens laparoscope. These models were validated, as the 0 degrees lens rigid laparoscope was surpassed by the 30 degrees lens rigid laparoscope and the flexible-tip laparoscopes. Both flexible-tip laparoscopes produced similar results and excellent image quality, but some experience is required before their smooth application can be achieved.

[A man with Candida pyopneumopericarditis and cardiac tamponade in conjunction with gastric tube infection]. / Een man met candida-pyopneumopericarditis en harttamponnade bij een buismaaginfectie.

A 55-year-old man who had undergone oesophagectomy with retrosternal gastric tube reconstruction for oesophageal carcinoma several years before, presented with retrosternal pain, fever and chills. He appeared to have Candida glabratarelated pyopneumopericarditis and a fungal infection in the gastric tube. Because of cardiac tamponade, the pericardium was surgically drained. The patient was given antibiotics and fluconazole. He left the hospital after one month in relatively good condition. Two months later, he was readmitted for haematemesis. During an emergency surgical procedure a fistula was found between the gastric tube and the left atrium. For these patients is early treatment of the underlying cause lifesaving. Monthly check-ups in an outpatient clinic are needed due to the risk of constrictive pericarditis and recurrent cardiac tamponade.

Skeletal metabolism in patients with osteoporosis after discontinuation of long-term treatment with oral pamidronate.

Bisphosphonates are used with increasing frequency in the treatment of patients with osteoporosis. Continuous administration of low doses of nitrogen-containing bisphosphonates by mouth is the preferred mode of therapy. The skeletal half-life of bisphosphonates is long, however, and little is known about their long term effects on skeletal metabolism. We examined the changes in biochemical parameters of bone turnover [serum alkaline phosphatase and urinary hydroxyproline (OHP)], in bone mineral density, and in fracture frequency after discontinuation of long term (mean, 6.5 yr, range, 5-9 yr) therapy with oral pamidronate (150 mg/day) in 30 patients with osteoporosis and vertebral fractures. Serum alkaline phosphatase and urinary OHP were significantly lower at the end of long term treatment (90% and 72% of basal values, respectively). Serum alkaline phosphatase had increased to basal values within 6 months of stopping treatment, whereas OHP increased significantly to a maximum average of 92% of pretreatment values. There was no change in the every 6-month bone mineral density measurements of the lumbar spine and the femoral neck during the 2 yr after stopping treatment. Spine fracture index, calculated by the method of Raymakers and co-workers, was 0.83 +/- 0.12 before treatment, 0.85 +/- 0.12 at the end of treatment, and 0.85 +/- 0.13 2 yr after stopping treatment (nonsignificant). There was also no significant change in the rate of new vertebral fractures on or up to 2 yr after stopping treatment (48.5 of 1000 and 46.5 of 1000 patient yr, respectively). Our data demonstrate that the sustained suppression of bone turnover induced by long term treatment with pamidronate is readily reversible on stopping treatment. The beneficial effect of this treatment regimen on the skeleton, however, appears to be maintained for at least 2 yr after discontinuation of treatment.

Recovery of serum calcium concentrations following acute hypocalcemia in patients with osteoporosis on long-term oral therapy with the bisphosphonate pamidronate.

Bisphosphonates, synthetic compounds that are taken up preferentially by the skeleton and that suppress bone resorption, are currently used in the management of patients with osteoporosis. Long-term uninterrupted administration of low oral doses is the preferred mode of treatment in current clinical trials with newer bisphosphonates. These compounds have, however, a long residence time in the skeleton, and there is no information about their long-term effects on blood calcium homeostasis. We examined the effect of long-term therapy with oral bisphosphonate on blood calcium homeostasis following an acute hypocalcemic stimulus. Twenty patients with vertebral osteoporosis (10 untreated controls and 10 treated with oral pamidronate, 150 mg/day for at least 5 yr) were given intravenous infusions of sodium EDTA, and the concentrations of calcium and PTH in blood were followed for 24 h. Serum calcium concentrations decreased similarly in both groups (maximum decrease 0.21 mmol/L and 0.22 mmol/L, respectively). The recovery of serum calcium concentrations was identical in both groups, and all patients had normal concentrations at 24 h. Plasma PTH increased to a peak of 17.3 +/- 2.5 pmol/L in the control group and to 17.0 +/- 3.1 pmol/L in the pamidronate-treated patients. During the whole study period, there was no difference in either the peak PTH response or in the recovery of plasma PTH values between the two groups. However, when only PTH responses between 60 min and 24 h were examined, there were differences between the two groups. Plasma PTH values, although strictly within the normal range, were significantly higher in the pamidronate-treated patients (P = 0.001). There were no differences in the calcemic responses during this period. Further, there were no detectable changes in immunoreactive PTH-related protein in either group after the EDTA infusions. In conclusion, our study showed that longterm therapy with oral pamidronate does not affect the calcemic response to an acute hypocalcemic stimulus in patients with osteoporosis.

The use of bisphosphonates in the treatment of osteoporosis.

The efficacy of bisphosphonates in the treatment of conditions characterized by increased osteoclastic bone resorption has been established. Recent evidence indicates that these compounds are also effective in the treatment of patients with osteoporosis. Two main protocols have been tried. One is based on the intermittent administration of the bisphosphonate, which is expected to decrease bone resorption, and give a drug-free period during which bone formation may proceed at a normal rate, leading to a positive calcium balance. The other argues that the resetting of the equilibrium in a cyclical process is, as a rule, incomplete and continuous low-grade suppression of resorption will result in a continuing positive bone balance. Intermittent administration of the first generation bisphosphonate, etidronate, for up to three years increases trabecular bone density, stabilizes it after two years, and appears to reduce the rate of new vertebral fractures in women with postmenopausal osteoporosis. Longer follow-up studies are needed before this beneficial effect is unequivocally established. Continuous administration of the second-generation bisphosphonate, pamidronate, increases spinal bone density in patients with osteoporosis linearly for up to four years, and is associated with a low rate of new vertebral fractures. These results need to be confirmed in controlled studies involving more patients. There are indications that pamidronate given continuously can prevent glucocorticoid-induced bone loss. There is no information about the effects of bisphosphonates on non-vertebral fractures. There are limited data about the use of bisphosphonates in the prevention of postmenopausal bone loss. Extensive studies on efficacy and safety are needed before this treatment is offered as an alternative to hormone replacement therapy.

Urinary telomerase and its possible role as a marker for bladder cancer.

Voided urine from patients with bladder cancer and from control patients with either hematuria or with no urologic conditions were examined for telomerase activity in order to explore the possibility that this activity could be used as a marker for the detection of bladder cancer. This assay was found to have an overall sensitivity in detecting bladder cancer of 85% (88/104) with 79% (23/29) grade 1 tumors, 84% (32/38) grade 2 tumors, 87.5% (28/32) grade 3 tumors, and 100% (5/5) carcinoma in situ testing positive. This compared favorably with urinary cytology which had an overall sensitivity of 51% and sensitivities of 13%, 44%, 82%, and 100% for grades 1, 2, 3 tumors and carcinoma in situ, respectively. The specificity of telomerase in patients with benign causes of hematuria was 66%, and in normal volunteers without urologic conditions, it was 100%. Assessment of nuclear matrix protein suggested comparable sensitivity and specificity. Evaluation of bladder tumor antigen showed less sensitivity for low-grade disease and less specificity, as it was influenced by inflammation and instrumentation. Telomerase thus seems to be a means whereby low-grade tumors may be detected in examination of voided urine and may offer an advantage in monitoring for recurrent disease.

Efficacy and safety of iopromide for excretory urography.

RATIONALE AND OBJECTIVES: Iopromide is a nonionic monomeric contrast agent. Initial laboratory and clinical data have shown that it is relatively safe. Efficacy for excretory urography has been shown to be good, comparable with other low-osmolality agents. The authors attempted to confirm these impressions in a randomized, double-blind comparison with equivalent doses of ioversol and iopamidol. METHODS: Two hundred adult patients undergoing excretory urography were studied. One hundred received iopromide, 40 received ioversol, and 60 received iopamidol (300 mg I/kg) as an intravenous bolus. Urographic films (obtained 1, 5, 15, and 20 minutes after the bolus, and postvoid) were interpreted by an observer blinded to contrast type. Visualization of renal parenchyma, pelvis and calyces, ureters, and bladder was independently assessed as excellent, good, poor, or nonvisualized. Vital signs were recorded before, 30 to 60 minutes after, and 24 hours after injection. Adverse reactions were sought, physical examinations were performed, and standard hematology and serum chemistry values were measured before and 1 day after injection; a 72-hour serum creatinine level was also measured. RESULTS: Ninety-eight percent of visualization scores were good or excellent; no significant differences among iopromide, iopamidol, and ioversol were found, nor were there any significant differences among groups in vital signs. Only one patient experienced a contrast-related physical examination change (subcutaneous extravasation). No significant changes with regard to hematology or serum chemistry values were observed; there was no contrast-induced nephropathy. Mild adverse reactions were experienced by 10% of patients; there were no significant differences in reaction rates among contrast agents. CONCLUSIONS: Iopromide at a dose of approximately 300 mg I/kg is safe and effective as an excretory urographic agent and is comparable in performance with ioversol and iopamidol.

Unrelated mismatched cord blood transplantation in patients with hematological malignancies: a single institution experience.

We report on six cases of unrelated UCB transplant in adult patients with hematological malignancies: three chronic myelocytic leukemias and three acute leukemias. Their median age and body weight were respectively: 28 years (range 15.5-40) and 55.5 kg (range 46-90). The cord blood units were from the New York Blood Center. The median number of nuclear cells provided, evaluated before thawing, was 2.1 x 10(7)/kg (range 1 x 10(7)/kg-4.7 x 10(7)/kg). The degree of HLA disparity was 1/6: two patients, 2/6: three patients, 3/6: one patient. The patients received a pretransplant regimen including total body irradiation. They were given graft-versus-host disease prophylaxis which consisted of cyclosporin A and corticosteroids. They were all given a combination of G-CSF and erythropoietin. The median time of white blood cell and platelet reconstitution were respectively 24 days (range 12-43) and 60 days (range 23-90). All the patients had a full chimerism. A grade I acute GVHD was observed in four patients and two patients do not have any GVHD. No chronic GVHD has been observed yet. Three patients died from toxicity. Three patients are alive and well in complete remission at 2 years, 1 year and 11 months post-graft.

Sensitivity and specificity of NMP-22, telomerase, and BTA in the detection of human bladder cancer.

OBJECTIVES: The recent introduction of novel molecular markers into clinical urology has created a need to evaluate the efficacy and utility of these potential markers. The ideal assay for bladder cancer should be noninvasive, sensitive, specific, and cost-effective. We compared the Matritech nuclear maxtrix protein (NMP)-22 assay, telomerase activity, and the Bard bladder tumor antigen (BTA) assay for the detection of human bladder cancer. METHODS: A single voided urine sample was obtained from patients with hematuria without bladder cancer and from patients with known bladder cancer before any treatment. Approximately 50 to 100 mL of voided urine sample was collected and aliquotted for the various assays. The results were compared to single cytologic results and ultimately to pathologic findings. RESULTS: In 47 patients with bladder cancer, the overall sensitivity was 81% for NMP-22, 80% for telomerase, 40% for BTA, and 40% for cytology. For Ta tumors (n = 31), sensitivity was 81% for NMP-22, 70% for telomerase, 32% for BTA, and 26% for cytology. For T1 or higher stage tumors (n = 13), sensitivity was 82% for NMP-22, 91% for telomerase, 64% for BTA, and 64% for cytology. The remaining 3 patients had carcinoma in situ (CIS). When tumors were stratified by tumor grade, grade I tumors (n = 16) were detected at 69% with NMP-22, 65% with telomerase, 13% with BTA, and 6% with cytology. Grade II tumors (n = 14) were detected at 86% with NMP-22, 72% with telomerase, 36% with BTA, and 36% with cytology. Grade III tumors (n = 14) were detected at 93% with NMP-22, 93% with telomerase, 79% with BTA, and 79% with cytology. Patients with CIS (n = 3) were detected at 67% with NMP-22, 100% with telomerase, 33% with BTA, and 67% with cytology. In 30 patients with hematuria but without bladder cancer, the overall specificity of the assays was 77% for NMP-22, 80% for telomerase, 73% for BTA, and 94% for cytology. CONCLUSIONS: In the population tested, NMP-22 and the telomerase assays gave similar sensitivity and specificity for the detection of bladder cancer, and appear to offer a greater sensitivity than the BTA assay and/or conventional cytology.