Recent breakthroughs demonstrate that peripheral diseases can trigger inflammation in the brain, causing psychosocial maladies, including depression. While few direct studies have been made, anecdotal reports associate urological disorders with mental dysfunction. Thus, we investigated if insults targeted at the bladder might elicit behavioral alterations. Moreover, the mechanism of neuroinflammation elicited by other peripheral diseases involves the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome, which is present in microglia in the brain and cleaves and activates proinflammatory cytokines such as IL-1β. Thus, we further explored the importance of NLRP3 in behavioral and neuroinflammatory changes. Here, we used the well-studied cyclophosphamide (CP)-treated rat model. Importantly, CP and its metabolites do not cross the blood-brain barrier or trigger inflammation in the gut, so that any neuroinflammation is likely secondary to bladder injury. We found that CP triggered an increase in inflammasome activity (caspase-1 activity) in the hippocampus but not in the pons. Evans blue extravasation demonstrated breakdown of the blood-brain barrier in the hippocampal region and activated microglia were present in the fascia dentata. Both changes were dependent on NLRP3 activation and prevented with 2-mercaptoethane sulfonate sodium (Mesna), which masks the effects of the CP metabolite acrolein in the urine. Finally, CP-treated rats displayed depressive symptoms that were prevented by NLRP3 inhibition or treatment with Mesna or an antidepressant. Thus, we conclude that CP-induced cystitis causes NLRP3-dependent hippocampal inflammation leading to depression symptoms in rats. This study proposes the first-ever causative explanation of the previously anecdotal link between benign bladder disorders and mood disorders.

Objective: To investigate the in vitro activation of the NLRP3 inflammasome within bladder urothelium by stone-forming components. Further, to describe the contributions of reactive oxygen species (ROS) and thioredoxin-interacting protein (TXNIP), an important structural component of the inflammasome, to this activation. Methods: Urothelial cells were harvested and incubated overnight. For agonist studies, cells were treated with varying concentrations of calcium pyrophosphate (CPPD) and monosodium urate (MSU). For inhibitor studies, cells were treated with either N-acetylcysteine (NAC) (1 hr) or Verapamil (4 hrs) prior to incubation with either CPPD (62.5 ug/mL) or MSU (1.25 ug/mL) for 24 hrs. Untreated controls were incubated with ATP (1.25 mM) for 1 hr to maximally stimulate NLRP3 inflammasome activity (measured as caspase-1 cleavage of the fluorogenic substrate Ac-YVAD-AFC). Results are reported as a percentage of maximum ATP response. Results: CPPD and MSU activate caspase-1 in urothelial cells in a dose-dependent manner, reaching~50% and~25% of the ATP response, respectively. Pre-treatment with the general ROS scavenger NAC reduces this activation in a dose-dependent manner. Additionally, activation was suppressed through treatment with Verapamil, a known downregulator of TXNIP expression. Conclusion: The stone components CPPD and MSU activate NLRP3 in an ROS and TXNIP-dependent manner in bladder urothelium. These findings demonstrate the importance of ROS and TXNIP, and suggest that targeting either may be a way to decrease stonedependent NLRP3 inflammation within the bladder.

Xylitol should not be used parenterally as a glucose substitute (banned in the USA).

No abstract

Patients requiring oral and/or enteral nutrition support for nutritional needs can form calcium oxalate (CaOx) kidney stones. Dietary oxalate, if excessive, can contribute to CaOx stones when unopposed by appropriate calcium. The oxalate concentration of oral/enteral nutrition formulas is not known. We assessed various formulas for oxalate.METHODS: Adult and pediatric oral/enteral nutrition formulas commonly used in hospitals as well as in home feeding regimens were selected. Formulas designed for oral and enteral consumption (or either) were included (table ); completely elemental (hydrolyzed) or modular formula products were not. Multiple samples (N, table) of each formula were acidified, heated, and centrifuged. Supernatants were filtered and analyzed for oxalate by ion chromatography. Oxalate concentration (mg/LAESD), relative standard deviation (SD) between samples (coefficient of variation; CV), and calcium:oxalate ratios (mg:mg/L of formula) were calculated.RESULTS: Of 35 formulas analyzed, 9 were excluded due to inconsistent results and high CVs. Results for the 26 remaining formulas are shown (table ). Oxalate concentration ranged from 4-140 mg oxalate/L of formula. Due to highly variable calcium content, calcium:oxalate ratios varied widely between formulas (from 0-286) with lower ratios suggesting higher potential for oxalate absorption. There was no difference between mean oxalate concentration of enteral vs. oral formulas (45 vs. 46 mg/L; P¼0.92). Formulas designated for enteral use tended to have lower relative SDs (mean CV 16% vs. 21% for oral formulas), likely due to the generally more complex matrix of oral formulas, which contributed to more analytical variability. Depending on the formula, a patient requiring 1.5 L daily could obtain anywhere from 12-150 mg oxalate.CONCLUSIONS: Patients requiring oral and/or enteral nutrition support are at risk for a high exogenous oxalate load depending on the formula ingested and on the bioavailability of oxalate. Patients with a history of or at high risk for urolithiasis would benefit from strategies to reduce the bioavailability of oxalate and urinary oxalate excretion, which may include supplemental calcium with feedings or use of an appropriate lower oxalate formula.

A chromosome analysis of the cells revealed a normal 46XY diploid karyotype. hBS11 cells expressed smooth muscle differentiation marker proteins, including MYH11, a-SMA, g-SMA, and calponin. The connexin 43 signals were also detected as spotty signals on the surface of hBS11 cells. M2 and M3 receptors expressed in differentiated hBS11 cells. Both the cholinergic receptor agonist calbachol and a high concentration of extracellular potassium increased intracellular calcium in differentiated hBS11 cells. Differentiated hBS11 cells stimulated by calcium ionophore (A23187) exhibited contractility.CONCLUSIONS: The present study demonstrates that the hBS11 cell maintains plasticity of the differentiation that a smooth muscle cell originally has. Immortalized hBS11 cell provides useful analytical system for elucidation of the pathophysiological mechanism of lower urinary tract dysfunction and the development of the new treatment options, in addition to the elucidation of molecular differentiation mechanism, reproduction mechanism, and the function control mechanism of the smooth muscle cell.