Radical cystectomy is the treatment of choice for muscle invasive bladder cancer and minimally invasive options have been explored to minimize the associated morbidity. Durak et al (page 1227) from New York, New York compared the surgical efficacy and efficiency of a completely suture based procedure with a novel entero-urethral anastomosis device and the EndoGIA™ to create an ileal neobladder. They investigated 2 groups of 7 pigs survived for 8 weeks. In group 1 neobladder construction was performed using a U-shaped segment of ileum and sealed with an EndoGIA. The entero-urethral anastomosis was created with a novel sutureless anastomosis device. All other procedures were completed with standard intracorporeal suturing techniques. Group 2 animals underwent a completely intracorporeal suture technique. Total procedure, and enteroenteric, ileal neobladder, ureteroenteric and entero-urethral anastomosis times were recorded. Immediate postoperative, 2-week and sacrifice cystograms were used to evaluate the newly constructed system.
In groups 1 and 2 the 2-week and sacrifice cystograms showed no extravasation. The overall surgical procedure, pouch creation and entero-urethral anastomosis times were statistically shorter for group 1. Average survival time in groups 1 and 2 was 30 (range 4 to 56) and 41 days (range 1 to 56), respectively. All animals developed voiding complications within 1 week after ureteral and urethral catheters were removed. The authors conclude that the combination of stapled construction of ileal neobladder and urethro-enteral anastomosis device significantly reduced the time required to complete the entire neobladder procedure and entero-urethral anastomosis. With continued refinement of these techniques laparoscopic, continent diversions may become a more clinically viable option.
Crucial Role of Interferon-γ in Experimental Autoimmune Prostatitis
Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is an important worldwide healthcare problem. It is a highly prevalent chronic inflammatory syndrome characterized by pelvic pain, irritative voiding symptoms and sexual dysfunction in young and middle-aged men. An autoimmune etiology has been proposed in cases that demonstrate interferon (IFN)-γ secreting lymphocytes specific to prostate antigens in the periphery and increased levels of IFN-γ in seminal plasma. Motrich et al (page 1213) investigated IFN-γ in nonobese diabetic and C57Bl/6 mice in which experimental autoimmune prostatitis (EAP) can be induced. They also studied IRF-1 and STAT-1 knockout mice deficient in transcription factors involved in IFN-γ signaling.
EAP is characterized by prostate specific IFN-γ secreting cells in the periphery and Th1 related cytokines in the target organ. The authors noted increased IFN-γ and interleukin (IL)-12 in prostates from autoimmune animals, and decreased or unaltered IL-10 and IL-4, respectively. Absence of transcription factors involved in the IFN-γ signaling cascade, eg IRF-1 and STAT-1, made mice resistant to EAP development. IRF-1 knockout and STAT-1 knockout mice immunized with prostate antigens did not have infiltration or alterations in the prostate. They did not demonstrate the typical prostate specific autoimmune response, and showed decreased IFN-γ, IL-12 and IL-10, and increased IL-4 in the prostate. The authors conclude that their results support a crucial role of IFN-γ in the pathogenesis of CP/CPPS, and emphasize that intense research is required to identify the etiopathogenic mechanisms underlying the disease and find a more rational therapy.
Over Expression of Chemokines and TNFSF14 From Ulcerative Interstitial Cystitis
Approximately 1 million individuals in the United States have interstitial cystitis (IC), of whom about 90% are women. There seems to be no consensus concerning the criteria for diagnosis, except for the ulcerative form of the disease. Ogawa et al (page 1206) from Japan investigated the genes responsible for IC by DNA microarray analysis and quantitative real-time polymerase chain reaction (RT-PCR). They defined patients by the 3 criteria of 1—lower urinary tract symptoms such as urinary frequency, bladder hypersensitivity and/or bladder pain, 2—bladder pathology proven endoscopically by Hunner's ulcer and 3—exclusion of confusable diseases such as urinary tract infection, malignancy or calculi. Bladder urothelial tissue was taken from a site apart from the ulcerative lesion in 9 patients with ulcerative IC and from normal looking areas in 9 controls (7 with bladder carcinoma and 2 with benign prostatic hyperplasia). Total RNA was extracted from the bladder samples and the expression levels of genes were compared between groups by microarray. Data were analyzed using GeneSpring™ GX software and Ingenuity Pathway Analysis, and chosen genes were confirmed for altered transcription by quantitative RT-PCR.
Using volcano plot analysis the authors identified 564 probes significantly expressed in mRNA more than 4-fold compared to the control group (p <0.001). By quantitative RT-PCR they confirmed the increased mRNA expression of several genes in the bladder samples from patients with ulcerative IC, including CXCR3 binding chemokines (CXCL9, 10 and 11) and TNFSF14 (LIGHT). Down-regulation of urothelial proteins such as uroplakins was also demonstrated. The authors conclude that genes related to immune and inflammatory responses, including Th1 related chemokines and cytokines such as CXCR3 binding chemokines and TNFSF14, are over expressed in bladder urothelium from patients with ulcerative IC. The over expressed genes and their products may give clues to the causative etiology of ulcerative IC and may be potential biomarkers for this disease.
Effect of Estrogen on Bladder Nociception
Numerous laboratory studies in animals clearly indicate a role for estrogen in nociceptive processing. In addition, clinical pains such as those associated with IC vary as a function of the menstrual cycle. Robbins et al (page 1201) from Birmingham, Alabama investigated the effect of estrogen on urinary bladder nociceptive sensation in an animal model previously demonstrated to be affected by the estrous cycle. Female Sprague-Dawley rats underwent ovariectomy or sham surgery and had visceromotor responses (VMRs, ie abdominal contractions) to bladder distention (UBD) 3 to 4 weeks later while anesthetized with isoflurane. In subsets of rats estrogen was either chronically replaced with a subcutaneous estrogen pellet vs placebo pellet or acutely replaced by subcutaneous injection 24 hours before testing. Estrogen withdrawal effects were examined in an additional group of rats by implanting a pellet and then explanting the pellet 24 hours before testing. Uterine weights were determined to assess estrogen dosing.
VMRs to UBD were significantly less vigorous in rats that underwent ovariectomy compared to controls. Acute estrogen replacement increased VMRs in these rats but chronic estrogen replacement did not. Sudden withdrawal of chronic estrogen resulted in increased VMRs. Uterine weight was consistent with the physiological estrogen dose. Estrogen alone was not sufficient to produce increased nociceptive responses but an acute decrease in estrogen resulted in increased VMRs. The authors emphasize that although the complexity of estrogen signaling is not completely understood, estrogens have genomic (direct control of gene expression) and nongenomic (regulation of cell signaling/phosphorylation cascades) effects that control critical cell signaling pathways. These effects have differing temporal characteristics with nongenomic effects tending to be rapid in onset and of limited duration whereas genomic effects are generally slower in onset and of more prolonged duration. The authors conclude that the overall effect of estrogen may depend on the balance between genomic and nongenomic signaling, central and peripheral effects, and receptor density.
Human Amniotic Fluid as New Source of Organ Specific Precursor Cells
Amniotic fluid (AF) is principally composed of fetal urine and lung exudates with a minor contribution from the amnion. AF contains novel stem cells (AFS), characterized by the expression of c-kit (stem cell factor receptor), which suggests that AFS may be useful in regenerative medicine. It has been shown that c-kit+ cells isolated from AF have the potential to differentiate into all 3 germ layers. Human AF contains multiple cell types, including pluripotent and committed, progenitor and fully differentiated cells. Da Sacco et al (page 1193) from Los Angeles, California characterized various cell populations in AF. Optimum culture techniques for multiple passages of cell lines with minimal morphological change were established. Analysis and characterization of these cell lines were performed using reverse transcriptase and RT-PCR. Immunoseparation was used to distinguish native progenitor cell lines and various subpopulations.
Endodermal and mesodermal marker expression was greatest in samples of early gestational age while ectodermal markers were expressed at a constant rate across all samples. Pluripotent and mesenchymal cells were always present while hematopoietic cell markers were expressed only in older samples. Specific markers for progenitor cells of the lung, kidney, liver and heart were increasingly expressed after 18 weeks of gestation. The authors specifically focused on a CD24+OB-cadherin+ population that can identify uninduced metanephric mesenchyma-like cells that are nephron precursors in vivo. The authors conclude that subpopulations of AF progenitor cells with renal characteristics could be a useful tool for therapy for various kidney diseases because of the commitment toward kidney cell types. The isolation of tubular and glomerular progenitors, particularly podocyte progenitors, may herald a novel approach to kidney regeneration compared to using pluripotential undifferentiated cells.
FULL TEXT