Immunological Status of Bladder Cancer Patients Based on Urine Leukocyte Composition at Radical Cystectomy

Background: Bladder cancer (BC) is the ninth most common malignancy worldwide, with high rates of recurrence. The use of urine leukocyte composition at the time of radical cystectomy (RC) as a marker for the study of patients’ immunological status and to predict the recurrence of muscle-invasive bladder cancer (MIBC) has received little attention. Methods: Urine and matched peripheral blood samples were collected from 24 MIBC patients at the time of RC. Leukocyte composition and expression of PD-L1 and PD-1 in each subpopulation were determined by flow cytometry. Results: All MIBC patients had leukocytes in urine. There were different proportions of leukocyte subpopulations. The expression of PD-L1 and PD-1 on each subpopulation differed between patients. Neoadjuvant chemotherapy (NAC), smoking status, and the affectation of lymph nodes influenced urine composition. We observed a link between leukocytes in urine and blood circulation. Recurrent patients without NAC and with no affectation of lymph nodes had a higher proportion of lymphocytes, macrophages, and PD-L1+ neutrophils in urine than non-recurrent patients. Conclusions: Urine leukocyte composition may be a useful tool for analyzing the immunological status of MIBC patients. Urine cellular composition allowed us to identify a new subgroup of LN− patients with a higher risk of recurrence.

Effectiveness of currently used urinary preservatives in preserving high demand biochemical analytes. A study in the context of Sri Lankan laboratory setting

Analysis of 24-hour urine composition is widely used in diagnosis of acute and chronic kidney diseases. In Sri Lanka, no guidelines are available on preservation of urine. This study aimed to identify an effective preservation technique/s among currently practiced techniques in laboratories to maintain chemical stability of high demand bio chemical analytes in a 24-hour urine collection. An experimental study was undertaken using urine samples from volunteers (n=42, National Hospital of Sri Lanka and University of Sri Jayawardenepura, Nugegoda, Sri Lanka). Each sample was divided in to 10 ml aliquots; Out of the lot, one was preserved without any preservative and another refrigerated at 4ºC for 24 hours. Other aliquots were preserved for 24-hours by addition of following preservatives in a 24-hour collection: boric acid g/ urine (5, 7.5, 10), sodium azide g/ urine (0.3, 0.6, 0.9), HCl ml/ urine (1N;10ml, 6N;10ml, 25ml and 30ml). The aliquots with preservatives were kept at room temperature for 24 hours. Protein, Creatinine, Ca2+, Mg2+ and PO43- concentrations were measured in each aliquot. Least mean squared error for each analyte in different preservatives was calculated. For protein and creatinine, least mean squared error was given by, 10g/L boric acid. That of Ca2+, Mg2+ and PO43 were observed when the sample was refrigerated or after addition of 10g/L boric acid as the preservative. Sodium azide and HCl that are utilized in current practice as preservatives for 24-hour urine collections do not show better performance in selected analytes. Boric acid (10g/L) is more effective in persevering protein and creatinine. Due to practical issues in acquiring refrigeration facilities, Ca2+, Mg and PO4 3- also can be preserved effectively by using the same preservative. Consequently, Boric acid (10g/L) could be recommended as an effective preservative to preserve selected analytes in 24-hour urine collection

Effect of dietary treatment and fluid intake on the prevention of recurrent calcium stones and changes in urine composition: A meta-analysis and systematic review

To perform a systematic review and meta-analysis of randomized controlled trials (RCTs) for investigating the effect of dietary treatment and fluid intake on the prevention of recurrent calcium stones and changes in urine composition. PubMed, Web of Science, Embase, EBSCO, and Cochrane Library databases (updated November 2020) were searched for studies with the following keywords: diet, fluid, recurrent, prevention, randomized controlled trials, and nephrolithiasis. The search strategy and study selection process was conducted by following the PRISMA statement. Six RCTs were identified for satisfying the inclusion criteria and enrolled in this meta-analysis. Our result showed that low protein with or without high fiber diet intervention does not decrease the recurrence of stone upon comparing with control groups (RR = 2.32, 95% CI = 0.42–12.85; P = 0.34) with significant heterogeneity among the studies (I2 = 81%, P = 0.02). But normal-calcium, low protein, low-salt diet had recurrences did reduced the recurrence compared to normal-calcium diet. And the fluid intake has a positive effect on prevention of recurrent stone formation (RR = 0.39, 95% CI = 0.19–0.80; P = 0.01) with insignificant heterogeneity among the studies (I2 = 9%, P = 0.30). The different components of urine at baseline were reported in four studies. Upon reviewing the low protein with or without high fiber dietary therapy groups, it was found that there were no obvious changes in the 24-hour urine sodium, calcium, citrate, urea, and sulfate. In conclusion, our study shows that the only low protein with or without fiber does not affect recurrence, but low Na, normal Ca diet has a marked effect on reducing recurrence of calcium stone. And fluid intake shows a significant reduction in the recurrence of calcium stone.

Composition of Urine Collected from Non-Stone-Forming Chinese Persons during Different Short-Term Periods of the Day

The purpose of this study in a small group of non-stone-forming Chinese persons was to measure the levels of supersaturation with calcium oxalate and calcium phosphate and pH with the aim of confirming if any of the different short-term urine samples were better for risk evaluation than a 24-h sample. Nine normal men and 1 woman collected urine during 4 periods of the day. Period 1 between 08 and 12 h, Period 2 between 12 and 18 h, Period 3 between 18 and 22 h, and Period 4 between 22 and 08 h. Each sample was analysed for calcium, oxalate, citrate, magnesium and phosphate, and estimates of supersaturation with calcium oxalate (CaOx) and calcium phosphate (CaP) were expressed in terms of AP(CaOx) and AP(CaP) index. An estimate of the solute load of CaOx was also calculated. Urine composition for 24-h urine (Period 24) was obtained mathematically from the analysed variables. Urine composition corresponding to 14-h urine portions 22–12 h (Period 14N) and 08–22 h (Period 14 D) were calculated. The lowest pH levels were recorded in Period 1 urine. The highest level of AP(CaOx) index was recorded during Period 1, and the product AP(CaOx) index × 107 × hydrogen ion concentration was significantly higher in Period 1 urine than in 24-h urine (p = 0.02). Also, the product SL(CaOx) × 107 × hydrogen ion concentration was significantly higher in Period 1 urine (p = 0.02). Low AP (CaP) index levels were recorded in Period 4, but also in all periods following dietary loads of calcium and phosphate. With the important reservation that the analytical results were obtained from non-stone-forming persons, the conclusion is that analysis of urine samples collected between 08 and 12 h might be an alternative to 24-h urine. The risk evaluation might advantageously be expressed either in terms of the product AP(CaOx) index × 107 × hydrogen ion concentration or the product SL(CaOx) × 107 × hydrogen ion concentration.

Sex Differences of Kidney Stone Urine Risk Factors after Roux-en-Y Gastric Bypass

<b><i>Introduction:</i></b> Roux-en-Y gastric bypass (RYGB) is a bariatric surgical procedure that is associated with higher risk of kidney stones after surgery. We examined urine composition in 18 men and women before and after RYGB to examine differences in kidney stone risk. <b><i>Methods:</i></b> Three 24-h urine collections were performed before and 1 year after RYGB. We analyzed mean urinary values for pre- and post-RYGB collections and compared men and women. <b><i>Results:</i></b> Seven men and eleven women completed pre- and post-RYGB urine collections. Pre-RYGB, men had higher calcium oxalate supersaturation (CaOx SS) (7.0 vs. 5.0, <i>p</i> = 0.04) compared with women. Post-RYGB, women had higher urine CaOx SS (13.1 vs. 4.6, <i>p</i> = 0.002), calcium phosphate supersaturation (1.04 vs. 0.59, <i>p</i> = 0.05), and lower urine volumes (1.7 vs. 2.7L, <i>p</i> &#x3c; 0.001) compared with men. <b><i>Discussion/Conclusion:</i></b> There are important differences in urine composition by sex that may contribute to higher kidney stone risk in women after RYGB compared with men.

A role for tubular Na+/H+ exchanger NHE3 in the natriuretic effect of the SGLT2 inhibitor empagliflozin

Inhibitors of proximal tubular Na+-glucose cotransporter 2 (SGLT2) are natriuretic, and they lower blood pressure. There are reports that the activities of SGLT2 and Na+-H+ exchanger 3 (NHE3) are coordinated. If so, then part of the natriuretic response to an SGLT2 inhibitor is mediated by suppressing NHE3. To examine this further, we compared the effects of an SGLT2 inhibitor, empagliflozin, on urine composition and systolic blood pressure (SBP) in nondiabetic mice with tubule-specific NHE3 knockdown (NHE3-ko) and wild-type (WT) littermates. A single dose of empagliflozin, titrated to cause minimal glucosuria, increased urinary excretion of Na+ and bicarbonate and raised urine pH in WT mice but not in NHE3-ko mice. Chronic empagliflozin treatment tended to lower SBP despite higher renal renin mRNA expression and lowered the ratio of SBP to renin mRNA, indicating volume loss. This effect of empagliflozin depended on tubular NHE3. In diabetic Akita mice, chronic empagliflozin enhanced phosphorylation of NHE3 (S552/S605), changes previously linked to lesser NHE3-mediated reabsorption. Chronic empagliflozin also increased expression of genes involved with renal gluconeogenesis, bicarbonate regeneration, and ammonium formation. While this could reflect compensatory responses to acidification of proximal tubular cells resulting from reduced NHE3 activity, these effects were at least in part independent of tubular NHE3 and potentially indicated metabolic adaptations to urinary glucose loss. Moreover, empagliflozin increased luminal α-ketoglutarate, which may serve to stimulate compensatory distal NaCl reabsorption, while cogenerated and excreted ammonium balances urine losses of this “potential bicarbonate.” The data implicate NHE3 as a determinant of the natriuretic effect of empagliflozin.