Calcineurin inhibitors (CNIs) are immunosuppressive drugs, which are used widely to prevent rejection of transplanted organs and treat autoimmune disease. Hypertension and renal tubule dysfunction, including hyperkalemia, hypercalciuria, and acidosis often complicate their use1,2. These side effects resemble familial hyperkalemic hypertension (FHHt), a genetic disease characterized by overactivity of the renal sodium chloride co-transporter (NCC), and caused by mutations in WNK kinases. We hypothesized that CNIs induce hypertension by stimulating NCC. In wild-type mice, the CNI tacrolimus caused salt-sensitive hypertension and increased the abundance of phosphorylated NCC, and the NCC regulatory kinases WNK3, WNK4, and SPAK. The functional importance of NCC in this response was demonstrated by showing that tacrolimus did not affect blood pressure in NCC knockout mice, whereas the hypertensive response to tacrolimus was exaggerated in mice over-expressing NCC. Moreover, hydrochlorothiazide reversed tacrolimus-induced hypertension. In kidney transplant recipients treated with tacrolimus, fractional chloride excretion in response to bendroflumethiazide was greater than in controls, and renal NCC abundance was also greater, extending these observations to humans. Together, these findings indicate that tacrolimus-induced hypertension is mediated largely by NCC activation, and suggest that inexpensive and well-tolerated thiazide diuretics may be especially effective in preventing the complications of CNI treatment.

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Introduction: Protein energy wasting is closely related to increased morbidity and mortality in peritoneal dialysis (PD) patients. Simple reliable and easily available methods of determining nutritional status and recognition of short-term changes in body composition are therefore important for clinical practice. Methods: We compared whole-body and segmental composition using multifrequency bioelectrical impedance analysis (MF-BIA) and dual-energy X-ray absorptiometry (DEXA) in 104 stable PD patients. Results: Assessment of whole-body composition showed that lean body mass (LBM) was highly correlated with good method agreement using DEXA as the reference test (r = 0.95, p < 0.0001; bias –0.88 kg, 95% CI –1.53 to 0.23 kg). Similarly, high correlation and good method agreement were found for fat mass (r = 0.93, p < 0.0001; bias 0.69 kg, 95% CI 0.03–1.36 kg). Segmental analysis of LBM revealed strong correlations between LBM for trunk, left and right arms and legs (r = 0.90, 0.84, 0.86, 0.89 and 0.90, respectively, p < 0.0001). Bone mineral content derived by MF-BIA overestimated that measured by DEXA (bias 0.740 kg, 95% CI 0.66–0.82 kg). Conclusion: MF-BIA may potentially be a useful tool for determining nutritional status in PD patients and serial estimations may help recognize short-term changes in body composition.

Cyclooxygenases, particularly COX-2, play an important role in tumor development and progression. We have previously shown that COX-2 expression is an independent prognostic factor in human ovarian carcinoma. In this study, we investigated the effects of the inhibition of COX isoforms by the NSAID NS-398 as well as by COXisoform-specific RNA interference (RNAi) in the human ovarian carcinoma cell lines OVCAR-3 and SKOV-3. OVCAR-3 cells showed a constitutive expression of COX-1 and an induction of high levels of COX-2 and PGE 2 after stimulation with interleukin-1b. In contrast, SKOV-3 cells were negative for both COX isoforms. In OVCAR-3 cells, PGE 2 production was inhibited by NS-398 in concentrations of 1 lm and by a COX-2-specific silencing RNA (siRNA), while a COX-1-specific siRNA did not have an effect. This suggests that COX-2 is the major source of PGE 2 in this cell line. To dissociate COX-2-specific and non-COX-2-specific effects on cell proliferation, a proliferation assay was performed after incubation of cells with NS-398 and COX siRNAs. NS-398 induced an inhibition of cell proliferation at concentrations of 50-500 lm, which are above the concentrations needed for the inhibition of PGE 2 production. This inhibitory effect was present in the COX-positive cell line OVCAR-3 as well as in the COX-negative cell line SKOV-3 and could not be reverted by addition of exogenous PGE 2 . Neither COX-1-nor COX-2-specific siRNAs had an effect on cell proliferation of OVCAR-3 cells. Cell cycle analysis showed an increased accumulation of cells in the G0/G1 phase after treatment with NS-398, but not with COX siRNAs. These experiments suggest that NS-398 reduced cell proliferation in ovarian carcinoma cells by induction of G0/G1 cell cycle arrest independent of COX-2 inhibition. Our study shows that specific inhibition of COX isoforms by RNAi could be used to dissociate effects of NSAIDs. Furthermore, our results suggest that cell cycle arrest is one of the primary mechanisms responsible for the antiproliferative effects of NS-398 on ovarian carcinoma cells.