TLR-2+ monocytes were reduced in Group 1 compared with Groups 2 and 3, and TLR-4+ monocytes were reduced in Groups 1 and 2 compared with Group 3. The frequencies and numbers of naïve CD4+ T and CD19+ B cells were higher in the three groups of neonates compared with adults, while

CD4+ effector and effector memory T cells and CD19+ memory B cells were elevated in adults compared with neonates, as expected. Our study provides reference values for leucocytes in cord blood from term and preterm newborns, which may facilitate the identification of immunological deficiencies in protection against extracellular pathogens. “
“CD28/B7 co-stimulation blockade with belatacept prevents alloreactivity in kidney transplant patients. However, cells lacking CD28 GSK458 manufacturer are not susceptible to belatacept treatment. As CD8+CD28− T-cells have MLN0128 purchase cytotoxic and pathogenic properties, we investigated whether mesenchymal stem

cells (MSC) are effective in controlling these cells. In mixed lymphocyte reactions (MLR), MSC and belatacept inhibited peripheral blood mononuclear cell (PBMC) proliferation in a dose-dependent manner. MSC at MSC/effector cell ratios of 1:160 and 1:2·5 reduced proliferation by 38·8 and 92·2%, respectively. Belatacept concentrations of 0·1 μg/ml and 10 μg/ml suppressed proliferation by 20·7 and 80·6%, respectively. Both treatments in combination did not inhibit each other’s function. Allostimulated CD8+CD28− T cells were able to proliferate and expressed the cytolytic and cytotoxic effector molecules granzyme

B, interferon (IFN)-γ and tumour necrosis factor (TNF)-α. Erastin manufacturer While belatacept did not affect the proliferation of CD8+CD28− T cells, MSC reduced the percentage of CD28− T cells in the proliferating CD8+ T cell fraction by 45·9% (P = 0·009). CD8+CD28− T cells as effector cells in MLR in the presence of CD4+ T cell help gained CD28 expression, an effect independent of MSC. In contrast, allostimulated CD28+ T cells did not lose CD28 expression in MLR–MSC co-culture, suggesting that MSC control pre-existing CD28− T cells and not newly induced CD28− T cells. In conclusion, alloreactive CD8+CD28− T cells that remain unaffected by belatacept treatment are inhibited by MSC. This study indicates the potential of an MSC–belatacept combination therapy to control alloreactivity. CD28/B7 co-stimulation blockade to prevent T cell activation and proliferation has been of interest for many therapeutic areas [1]. Belatacept, the latest immunosuppressive drug approved for therapy of kidney transplant recipients, utilizes this blocking mechanism. It is a fusion protein consisting of the extracellular domain of cytotoxic T lymphocyte antigen-4 (CTLA-4) and the Fc region of a human immunoglobulin (Ig)G1 immunoglobulin. By binding to CD80 (B7.1) and CD86 (B7.2) with a higher affinity than CD28, belatacept blocks the co-stimulatory signal [2].

16 of nine major mortality studies comparing PD and HD to investigate any trends in outcomes within selected subgroups of patients. Six large-scale registry studies and three prospective cohort studies were included in the analysis. The studies www.selleckchem.com/products/MG132.html included originated from the USA, Canada, the Netherlands and Denmark. The differences in study results were attributed to the amount of case-mix adjustment made and the subgroup

investigated. When these differences were accounted for, the critical review cited a remarkable degree of synergism in results. Peritoneal dialysis was generally found to have equal, if not better, survival in younger diabetic and non-diabetic patients regardless of study origin; however, there were variations in results with the older diabetic population. Only in the United States was there shown to be a survival advantage for the older diabetic patient to choose HD therapy

over PD. All studies demonstrated a time-dependent trend in the RR of death. All studies associated PD with equivalent or better survival during the 2 years of dialysis. Survival outcomes based on dialysis modality have been heavily researched internationally with the larger registry data-based studies dominating publications, most of which are from the United States and the Netherlands. It is important to review the more recent publications when assisting with patient modality choice as the survival trends of American patients on PD have shown double the improvement in survival rates when compared with HD survival improvement in the past few years. When analysing more recent patient populations with clearer dialysis p38 MAPK pathway Dichloromethane dehalogenase adequacy targets, we are able to identify that PD therapy is at least equivalent to HD therapy overall, but when considering subgroups such as age, diabetes and CVD, survival differences do become apparent. There has been one randomized controlled trial by Korevaar et al.7 in the Netherlands,

which needs to be interpreted with caution. Only 38 patients were recruited to this trial, which ceased early due to a lack of participants. At least 100 patients were needed to provide statistical power. There was some modality switching given the ethical and logistical difficulties of running a randomized controlled trial in this area. However, there was a significant survival benefit to those commencing on PD at least in the 4-year follow up, which was consistent, although less prominent, even after adjustment for the modality switching. The majority of the studies investigating mortality associated with modality are cohort or registry data studies. These publications do differ according to their criteria for inclusion; incident versus prevalent patient populations; intention-to-treat versus as-treated models; duration of follow up; varying adjustments for comorbidity number and severity; and subgroup analysis.

Overall, there is stronger molecular evidence that IL-2 is important for Th2 generation 23 than for Th1 cells. This leaves open the possibility that a direct Th2-skewing effect of IL-2 may also contribute to the protective function of IL-2-antibody complexes in myasthenia gravis. Although recent interest in IL-2 for the treatment of autoimmunity stems from IL-2′s role in the maintenance of Treg, the activities of this growth factor on effector cells must be taken into account when evaluating IL-2′s therapeutic potential. Indeed, in a study using IL-2 to prevent diabetes in CYC202 ic50 the NOD mouse 16, it was found that high doses

of IL-2 complexes expanded effector cells and led to accelerated onset of diabetes. To the contrary, low doses of IL-2 prevented the onset of diabetes by restoring functional Treg in the pancreas. The detrimental outcome www.selleckchem.com/products/dorsomorphin-2hcl.html of IL-2 treatment is likely due to the induction of strong effector and memory responses in the autoimmune-prone mice. Recent studies have demonstrated a prominent role for IL-2 in

the generation of CD4+ and CD8+ memory T cells 24, 25. This function is obviously unwanted when treating autoimmune disease but the current body of evidence suggests that careful determination of IL-2 complex doses and timing to specifically target Treg provides a rationale to circumvent these problems 16. It may be that lower doses of IL-2 complexes favor Treg function, whereas higher doses will boost effector/memory cell formation. The high levels of CD25 on Treg compared with naïve and effector cells may explain this differential sensitivity, allowing Treg to outcompete effector cells for capturing

environmental IL-2. It is likely that assays for IL-2 action, such as phospho-flow analyses 14, will help us better define IL-2′s targets under different conditions of exposure. In addition, combination therapy, such as IL-2 to promote Treg numbers and function and mTOR inhibitors to block the generation of effector T cells, may prove to be beneficial in immunological disorders. IL-2 is one of the first cytokines G protein-coupled receptor kinase discovered and it was thought that IL-2′s function is well understood. Studies in the past 10 years have led to new insights into the biology of IL-2 and an astonishing re-evaluation of the dogma. It is especially remarkable that almost two decades ago this cytokine was being tested for its ability to boost immune responses and now it is being considered as a therapy to inhibit immune responses. The development of better assays to define cytokine actions in vivo and rational strategies to optimize the actions of cytokines may help to realize the potential of IL-2 as an immunotherapeutic agent. Supported by NIH grants RO1 AI073656 and P01 AI35297 (to A. K. A.), and JDRF grant 32-2008-354 (to H. D.). Conflict of interest: The authors declare no financial or commercial conflict of interest. See accompanying article: http://dx.doi.org/10.1002/eji.

One-way ANOVA was used as appropriate to analyze rER variances of areas (I, O, and C) within each survival group. Differences between individual bone forming areas within samples were analyzed with paired t-tests. Differences between isotransplants and allotransplants and between survival periods were compared

with unpaired t-tests. Data are presented as mean ratio with standard deviation. Significance is click here set at p < 0.05. In all animals, the pedicle was patent at inspection of polymer filling of the vasculature. The rER in allotransplants at 4 weeks (A) was 0.456 ± 0.266 in the overall cortical area, while it was slightly higher at the outer cortex; 0.549 ± 0.184 and lower at the inner cortex; 0.362 ± 0.081. The rER at 18 weeks (group B) had increased in all areas, with an overall cortical rER of 0.749 ± 0.387; however, this difference did not reach significance (p > 0.05). The rER Antiinfection Compound Library at the inner cortex at 18 weeks was 0.532 ± 0.188, at the outer cortex 0.586 ± 0.175 (Table

1). In the isotransplant group at 4 weeks (group C), the overall cortical rER was 0.412 ± 0.239. The inner cortex had a rER of 0.398 ± 0.241, while at the outer cortex the rER was 0.247 ± 0.181. At 18 weeks in isotransplants (group D), the overall rER was slightly higher 0.467 ± 0.252 than group C (p > 0.05), with an inner cortex rER of 0.356 ± 0.113 and an outer PtdIns(3,4)P2 cortex rER of 0.392 ± 0.229. The short-term survival groups (A and C) had a comparatively equal overall cortical rER. At 18 weeks, the rER was higher in allotransplants (group B) as compared to the isotransplants (group D); however, no statistical significant difference was found (p > 0.05). At the outer cortical areas, the rER was significantly lower at 4 weeks in isotransplants

as compared to allotransplants (p < 0.05). This difference at the outer cortex was not found at 18 weeks. In the allotransplant group, a slight increase over time was found at the inner cortex, while in isotransplants, the rER remained lower than 0.5 over time with a majority of cells of donor origin. For successful incorporation and optimal biological properties of bone grafts, remodeling is a prerequisite. To understand the biology behind this process, knowledge of cellular heritage and the movement of cells in the transplant over time is essential. We applied a sex-mismatch rat model that has been used successfully in our previous bone transplantation research.[15-17] This transplantation model allows the study of cell lineage with quantitative RT-PCR on the Sry and cyclophilin housekeeper genes to detect the relative amount of recipient cells to donor cells within the transplant. Laser capture microdissection facilitates highly selective harvest of tissue, without contamination of adjacent soft tissue including capillary tissue.

maxima APGA could induce cross-protection

to heterologous species, E. tenella and E. acervulina, as well as E. maxima infections (68,69). At this point, it was believed that this cross-reactivity was most probably due to conserved epitopes of the major gametocyte antigens in the different species and, thus, led to the hypothesis that a vaccine of E. maxima gametocyte antigens could possibly be used to control coccidiosis caused by at least the three predominant Eimeria species. In floor pen maternal immunization trials, the resistance of chicks from APGA-immunized breeder hens was compared to that of hatchlings from control parent flocks by introducing oocysts to the pens via infection of ‘seeder’ birds infected with 50 oocysts of E. maxima, Fulvestrant clinical trial E. tenella and E. acervulina. Analogous to the laboratory trials, a reduction of 60–80% in oocyst shedding into the litter was observed; comparable to the reduction observed using coccidiostats (59). These NVP-LDE225 nmr trials were repeated three times under the same conditions, showing that there was an average reduction of 60–70% oocyst output in vaccinated groups up to 4–6 weeks in age (59,70). These results were encouraging, as they supported the idea that the highly conserved E. maxima antigens could provide good levels of protective immunity against at least three major species

that cause coccidiosis in broilers. Despite this, questions still remained about whether this vaccine could provide maternal protection against all Eimeria species (and strains) encountered in the field and if maternal immunity was applicable in controlling coccidiosis

within industry management schemes and climatic conditions. To further assess the efficacy of maternal immunization with APGA, testing was undertaken in a multi-centred, multinational field trial involving five countries from four different continents: Israel, Brazil, Argentina, South Africa and Thailand (71,72). The safety and immunogenicity of the vaccine in breeding hens were assessed on a large scale, with birds vaccinated twice prior to the start of their laying period (15 and 20 weeks respectively). Immunizations were found to have no deleterious effect on the hens (72): no adverse reactions; no damage at the site of injection; and no affect on hen mortality or on the number C-X-C chemokine receptor type 7 (CXCR-7) of eggs produced (e.g., in the Argentine trials, 119 eggs were produced per immunized hen vs. 116 per control hen). In all four countries, IgG antibody titres remained at a presumptive protective level throughout the life of the laying hens. The maintenance of highly specific IgG antibody levels in vaccinated flocks is thought to be due to the boosting effect that is naturally acquired from exposure to infection with oocysts in the environment (72). It is even conceivable that maternal antibodies may increase due to this natural exposure. However, in the absence of immunization, these titres are variable and, therefore, do not necessarily provide protective levels of maternal immunity (72).

8 pg/mL, which is similar to that of EHEC-derived Stx2 (2.5 pg/mL), whereas the CD50 of mStx2-His was considerably higher (585 ng/mL). On the other hand, the intraperitoneal MLD of Stx2-His in adult mice (6 weeks of age) was 100 ng, whereas that of mStx2-His was considerably higher (100 μg), indicating that the activities of these mutant toxins are close to non-hazardous when administered

at vaccination dosages. To confirm the effect of mStx2-His as a vaccine antigen, we immunized HM781-36B price ICR mice s.c. with 10 μg of mStx2-His containing aluminum hydroxide as a practical adjuvant for vaccine. No mice died of or were weakened by the immunization. As shown in Figure 3a, the IgG antibody titers in mice that were immunized twice with mStx2-His were significantly higher (mean ± SEM 2,206,250 ± 335,643, range 156,250–3,906,250) than those of mice immunized with adjuvant alone (titers of all five were < 10). The neutralizing activities of these antibodies were confirmed by an in vitro neutralization assay using 10 pg/mL of EHEC-derived Stx2 (corresponding to a 20.9% survival concentration in HeLa229 cells). No sera derived from

immunized mice with PBS neutralized the toxicities (mean ± SEM of survival rate 25.7 ± 0.4%), whereas the sera derived from immunized mice with mStx2-His neutralized the toxicities (mean ± SEM survival rate 70.3 ± 7.0%). To investigate the degree of protection Carfilzomib cell line conferred by antibodies that were induced in mice by immunization with mStx2-His, we divided the mice into three groups and challenged them with different lethal doses of wild-type Stx2. In this

study, we used Stx2-His to challenge mice with high lethal doses of purified toxin on the assumption that a large amount of toxin protein was needed. As shown in Figure 3b, all the mice immunized with mStx2-His survived a challenge of Stx2-His at 10- and 100-fold MLD (1 and Demeclocycline 10 μg/mouse, respectively) for at least 1 week with no symptoms, whereas only three of nine mice survived a challenge of 1000-fold MLD (100 μg/mouse). All of the mice immunized with adjuvant alone succumbed to a challenge with 10-fold MLD within 3 days. It is crucial to consider the following three points if toxoids are to have clinical utility. First, the toxoid itself must not be hazardous to humans and animals. Second, the toxoid should induce sufficient antibody production to neutralize an excess amount of wild-type toxin. Third, it should be possible to prepare large amounts of the toxoid antigen easily and cheaply. Taken together, these factors necessitate use of the overexpression method for preparation of antigenic proteins. In the process of constructing the CTB expression plasmid in our previous study [25], we confirmed that the SD sequence derived from LTB worked well for expression of the Vibrio cholerae derived CTB gene in E. coli without any obvious toxicities.

An internal standard is used to ensure precision in mass determination. The result is that the Ibis

universal biosensor detection system can identify the amplicons produced by a carefully designed primer set, with a high degree of accuracy that is stated as a percentage in the ‘read out’ data and with a sensitivity that detects all organisms present as >1% of the total microbial population in the sample. The system also detects and identifies fungi and viruses, and detects the presence of the bacterial genes that control resistance to antibiotics. Primer sets can be designed buy Y-27632 to focus on the pathogens usually seen in a particular medical situation, such as orthopedic infections, so that sensitivity and accuracy can be enhanced in the parts of the bacterial ‘tree of life’ (Fig. 5) in which the majority of the ‘usual suspects’ are located. The time required for DNA extraction is short, except in exceptional cases, and the PCR amplification process is rapid and automated, so that the Ibis system can detect and identify all of the bacteria present in a sample in <6 h, and biofilm cells are detected with the same sensitivity as planktonic cells. We have initiated prospective

double-blinded studies of both suspected infections of total joint prostheses, and of infected nonunions of the tibia/fibula following open trauma, in which we will compare data obtained from cultures with data generated using the Ibis system. Clinical decisions will be based on culture data because the Ibis system is not yet FDA approved, but after the code has been broken, buy Crizotinib the sensitivity and accuracy of the Ibis system will be compared with that of cultures. In addition, the Ibis data will be considered retrospectively, as a potential basis for clinical decisions, in the light of clinical outcomes and in the light of additional evidence of the presence of bacterial biofilms, such as direct microscopic evidence using FISH probes. If Amino acid the sensitivity and accuracy of the Ibis system are seen to exceed those

of traditional cultures, we will support their adoption for the diagnosis of bacterial infections in all aspects of orthopedic surgery. “
“Here, we report on the successful programming of dendritic cells (DCs) using selectively applied mixtures of chemokines as a novel protocol for engineering vaccine efficiency. Antigen internalization by DCs is a pivotal step in antigen uptake/presentation for bridging innate and adaptive immunity and in exogenous gene delivery used in vaccine strategies. Contrary to most approaches to improve vaccine efficiency, active enhancement of antigen internalization by DCs as a vaccine strategy has been less studied because DCs naturally down-regulate antigen internalization upon maturation. Whereas chemokines are mainly known as signal proteins that induce leucocyte chemotaxis, very little research has been carried out to identify any additional effects of chemokines on DCs following maturation.

Echinocandins are characterised by a good safety profile, few drug–drug interactions and good susceptibilities. With the increase in potentially azole-resistant non-albicans infections, echinocandins may become the first-line treatment of choice for many patients. “
“We report a case of non-fatal disseminated Scedosporium prolificans infection, including central nervous system disease and endophthalmitis, in a relapsed acute myeloid leukaemia patient with extensive CYP2C19 metabolism. Successful treatment required aggressive surgical debridement, three times daily voriconazole dosing and cimetidine CYP2C19 inhibition.

In addition, the unique use of miltefosine was employed due to azole-chemotherapeutic drug interactions. Prolonged survival following disseminated S. prolificans, adjunctive miltefosine and augmentation of voriconazole exposure with cimetidine CYP2C19 inhibition has not been reported. Selumetinib in vitro “
“The extensive use of immunosuppressive therapies in recent years has increased the number of patients prone to GDC-0449 cost or actually suffering from localised candidosis. As Candida species gain increasing resistance towards common antifungal drugs, new strategies are needed to prevent and treat infections caused by these pathogens. Probiotic bacteria have been in vogue in the past two decades. More and more dairy products containing such organisms offer

promising potential beneficial effects on human health and well-being. Because of the ability of probiotic bacteria to inhibit the growth of pathogens and to modulate Rebamipide human immune responses, these bacteria could provide new possibilities in antifungal therapy. We summarise the recent findings concerning the usefulness of probiotic treatment in localised candidosis, as well as discussing possible risks of probiotic treatment and highlighting the

molecular mechanisms that are believed to contribute to probiotic effects. “
“The fungal pathogen Candida albicans is a leading causative agent of death in immunocompromised individuals. Many factors have been implicated in virulence including filamentation-inducing transcription factors, adhesins, lipases and proteases. Many of these factors are glycosylphosphatidylinositol-anchored cell surface antigenic determinant proteins. Pga1 is one such protein shown to be upregulated during cell wall regeneration. The purpose of this study was to characterise the role Pga1 plays by creating a homozygous pga1 null strain and comparing the phenotype with the parental strain. It was observed that the mutant strain showed less oxidative stress tolerance and an increased susceptibility to calcofluor white, a cell surface disrupting agent that inhibits chitin fibre assembly which translated as a 40% decrease in cell wall chitin content. Furthermore, the mutant exhibited a 50% reduction in adhesion and a 33% reduction in biofilm formation compared with the parental strain, which was reflected as a slight reduction in virulence.

Even though it appeared as the HBD1 and HBD3 mRNA expression was down-regulated by Th2 cytokines and histamine, no statistical differences were found (Fig. 4a–c). Moreover, high levels of HBD1-3 were excreted from tonsils, but the levels remained unchanged upon stimulation (Fig. 4d–f). However, our impression was that the outcome of these Tyrosine Kinase Inhibitor Library cell line analyses

was dependent on where the excised tonsillar piece was taken. It was technically very difficult to know in advance the relation between epithelial and lymphoid cells as well as the infectious and allergic status of the tonsil and donor, respectively. Therefore, the experiments were repeated with mixed tonsillar lymphocytes and AECs cultured for 4, 16 and 24 h with and without IL-4, IL-5 and histamine. For both cell types, 4 and 16 h were insufficient to induce AMP generation (data not shown). However, after 24 h of culture the effects on the lymphocyte-induced HBD release were negligible (Fig. 5a–c), whereas a marked reduction in the epithelium-derived HBDs in the culture medium was seen in response to these agents (Fig. 6a–c). The present study describes HBD1-3 in tonsillar tissue and their regulation

in allergic rhinitis. mRNA and protein expression of HBD1-3 are shown in epithelial and lymphoid cells along with tonsillar secretion of HBD1-3. Allergic individuals are found to have reduced levels of HBD1-3. In addition, culture of mixed tonsillar lymphocytes and AECs with Th2-associated

cytokines and histamine causes a down-regulation of HBDs in the latter, indicating that the epithelial tissue is the regulatory site for the production of HBDs. Respiratory infections are Dinaciclib clinical trial known to cause exacerbations of allergic disease. AMPs, including HBDs, are key players in the first line defense against such infections. The present study demonstrates the presence of HBD1-3 in tonsils and that they originate from the epithelium as well as CD4+, CD8+ and CD19+ lymphocytes. Presence of AMPs in tonsillar tissue as well as their association with airway infections has previously been thoroughly described (Ball et al., 2007; Schwaab et al., 2009). Tieu et al. (2010) have investigated 4-Aminobutyrate aminotransferase members of the S100 family in chronic rhinosinusitis, and reported diminished levels of epithelial psoriasin (S100A7) and calprotectin (S100A8/A9). In analogy, reduced mRNA levels of psoriasin have been observed in infected tonsils (Bryborn et al., 2008). Claeys et al. (2003) have demonstrated high levels of mRNA encoding HBD2 and HBD3 in tonsils with no significant difference between idiopathic hypertrophic tonsillar disease and recurrent tonsillitis. Another group found presence of HBD1-3 in tonsils and that the concentrations were similar during different states of tonsillar disease (Schwaab et al., 2010). The reduced HBD1-3 levels found in tonsils from AR patients are in line with previous studies reporting a reduction in AMP synthesis in allergic individuals.

Moreover, an extra long segment of bowel is not required (unlike Studer’s), and the procedure is versatile and not technically difficult.[9] Urodynamic evaluation

is the most objective method in assessing the functional outcome of a neobladder. Various authors have addressed the urodynamic behavior of neobladder including urethral closure characteristics.[2, 5, 7] This study was conducted to evaluate short-term functional, urodynamic and QOL outcomes of our early experience with “W” ileal orthotopic neobladder (ONB), with non-refluxing extramural serosa-lined tunnel uretero-ileal anastomosis. Consecutive men undergoing cystoprostatectomy and ONB during December 2009 to March 2011 were enrolled. The study was approved by the institute’s ethical RG7204 order committee. The protocol was explained to each participant and written informed consent was taken prior to inclusion. The orthotopic neobladder (ONB) reconstruction

was fashioned using an ileal segment made into a W configuration and serous-lined ureteral reimplantation (Ebol Eneim and Ghoneim).[10] Patients were put on clean intermittent catheterization under the following circumstances: (i) For pouch wash – initiated twice a week in early postoperative period and progressively tapered; (ii) periodic self-calibration after endoscopic management of stricture; (iii) bothersome lower urinary tract symptoms (LUTS) with high post-void residual urine (PVR). Evaluation included: Neobladder pouch-related quality of life evaluation (PQOL) modified from study by Gotoh et SCH772984 molecular weight al.[11] The questionnaire consisted of 31 questions (total scoring 31–120; high score is more adverse),

divided into five domains: (i) Urine evacuation domain (11 questions; scoring 11–43); (ii) urine storage domain (13 questions; scoring 13–48); (iii) micturition status domain (two questions; Progesterone scoring 2–8);(iv) limitation in daily life domain (three questions; scoring 3–15); (v) pain domain (two questions; scoring 2–6). Urodynamics (Solar silver, MMS International, Enschede, The Netherlands):(i) Free uroflowmetry – standing posture; (ii) filling and voiding cystometry (pouchometry) – sitting posture; (iii) resting urethral pressure profilometry – sitting posture; (iv) surface electromyography; (v) filling and voiding poucho-urethrography – filling in supine and voiding in standing posture. Three-lumen urethral pressure profilometry catheter (8Fr, Rusch, Germany) and single lumen balloon rectal catheter (5Fr, Medica, Medolia, Italy) were placed. Catheters were “zeroed” to atmospheric pressure keeping the transducers at the level of the superior border of pubic-symphysis. Sterile normal saline (0.9%, w/v) was used as the filling medium and infused at a rate approximately 10% of functional pouch capacity (based on bladder diary) via the urethral catheter using a motor-driven and computer-controlled infusion-pump.