Anti-TLR7 Antibody Protects Against Lupus Nephritis in NZBWF1 Mice by Targeting B Cells and Patrolling Monocytes.

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by autoantibody production and multiple organ damage. Toll-like receptor 7 (TLR7), an innate immune RNA sensor expressed in monocytes/macrophages, dendritic cells (DCs), and B cells, promotes disease progression. However, little is known about the cellular mechanisms through which TLR7 drives lupus nephritis. Here, we show that the anti-mouse TLR7 mAb, but not anti-TLR9 mAb, protected lupus-prone NZBWF1 mice from nephritis. The anti-TLR7 mAb reduced IgG deposition in glomeruli by inhibiting the production of autoantibodies to the RNA-associated antigens. We found a disease-associated increase in Ly6Clow patrolling monocytes that expressed high levels of TLR7 and had upregulated expression of lupus-associated IL-10, CD115, CD31, and TNFSF15 in NZBWF1 mice. Anti-TLR7 mAb abolished this lupus-associated increase in patrolling monocytes in the circulation, spleen, and glomeruli. These results suggested that TLR7 drives autoantibody production and lupus-associated monocytosis in NZBWF1 mice and, that anti-TLR7 mAb is a promising therapeutic tool targeting B cells and monocytes/macrophages.

A Prospective Multicenter Randomized Controlled Study on Interleukin-6 Removal and Induction by a new Hemodialyzer With Improved Biocompatibility in Hemodialysis Patients: A Pilot Study.

We compared interleukin-6 (IL-6) removal and induction between conventional polysulfone (Con) and TORAYLIGHT NV (NV) dialyzers in hemodialysis patients. Twenty patients on Con with high IL-6 concentrations (2.7-8.5 pg/mL) were randomized to Con or NV group. Dialyzer performance was determined in NV group while patients were on Con and after being switched onto NV. Erythropoiesis-stimulating agent (ESA) response index (ERI) was assessed every 4 months for one year. IL-6 clearance was comparable between Con and NV. IL-6 removal rates were comparable for the first 1 h, but were higher with NV for the entire session (P = 0.03). Before-to-during-dialysis IL-6 concentration ratios were lower with NV on the venous side after the session (P = 0.03). During the one-year study, hemoglobin was lower in Con group than in NV group at month 8 (P = 0.046). ERI decreased in NV and increased in Con group, with a significant difference between the groups (P = 0.002). NV and Con are comparable in removing IL-6 and both induce IL-6. However, the data suggest that NV induces less IL-6, which may reduce the risk of ESA hyporesponsiveness.

Effect of sevelamer and calcium-based phosphate binders on coronary artery calcification and accumulation of circulating advanced glycation end products in hemodialysis patients.

BACKGROUND: Some trials have indicated that coronary artery calcification progresses more slowly in sevelamer-treated dialysis patients than in those using calcium-based binders. Effects of phosphate binders on circulating advanced glycation end products (AGEs) are unknown. STUDY DESIGN: Randomized trial with parallel-group design. SETTING & PARTICIPANTS: 183 adult (aged >20 years) patients on maintenance hemodialysis therapy at 12 dialysis facilities with a mean vintage of 118 ± 89 (median, 108) months. Dialysate calcium concentration was 2.5 mEq/L, and dietary calcium was not controlled. INTERVENTION: Patients were randomly assigned to 12 months of treatment with sevelamer (n = 91) or calcium carbonate (n = 92). OUTCOMES & MEASUREMENTS: Primary outcome measures were change from baseline in coronary artery calcification score (CACS) determined at study entry and completion using multislice computed tomography and the proportion of patients with a ≥ 15% increase in CACS. Blood parameters were determined at study entry and 2-week intervals, and levels of plasma pentosidine, a representative AGE, were determined at study entry, 6 months, and study completion. RESULTS: 79 (86.8%) and 84 (91.3%) patients in the sevelamer and calcium-carbonate arms completed the treatment, respectively. Both binders were associated with an increase in mean CACS: 81.8 (95% CI, 42.9-120.6) and 194.0 (139.7-248.4), respectively (P < 0.001 for both). After adjustment for baseline values, the increase in the sevelamer group was 112.3 (45.8-178) less (P < 0.001). Percentages of patients with a ≥ 15% increase in CACS were 35% of the sevelamer group and 59% of the calcium-carbonate group (P = 0.002). Plasma pentosidine levels increased with calcium carbonate but not [corrected] sevelamer treatment (P < 0.001). Sevelamer use was associated with decreased risk of a ≥ 15% increase in CACS regardless of baseline blood parameters, pentosidine level, and CACS. LIMITATIONS: Treatment duration was relatively short, some sevelamer-treated patients (7 of 79) received calcium carbonate, and washout could not be performed. CONCLUSIONS: The data suggest that sevelamer treatment slowed the increase in CACS and suppressed AGE accumulation.

Can cinacalcet replace parathyroid intervention in severe secondary hyperparathyroidism?

A 6-month observational study was conducted in 61 patients (33 men and 28 women, mean age 54.8 +/- 12.4 years) treated with cinacalcet in whom parathyroid intervention was indicated. Thirty-seven patients had baseline intact parathyroid hormone (iPTH) levels of >500 pg/mL, but only five still had levels this high after 6-month cinacalcet therapy. No patients had phosphorus (P), calcium (Ca), or iPTH levels within the target range at baseline, but six patients (9.8%) reached all three target ranges after treatment. The stratum with many patients who had 2-4 enlarged parathyroid glands shifted toward the low PTH groups (iPTH < 300 pg/mL) with treatment. There was less of a tendency for patients with more enlarged glands, that is, 10 mm or larger at baseline, to have a higher PTH level after cinacalcet treatment. There was no significant difference in the total volume of parathyroid glands after treatment, since some glands enlarged while others shrank. These findings indicate cinacalcet to be a potentially useful treatment. Our results suggest that 80% of cases indicated for parathyroid intervention could avoid such interventional therapies with cinacalcet administration. However, the variability in the gland-shrinking effect of cinacalcet on parathyroid glands merits further study.

Suppression of parathyroid hormone production in vitro and in vivo by RNA interference.

We used RNA interference, which causes sequence-specific degradation of target mRNAs to suppress the production of parathyroid hormone by cells of patients with secondary hyperparathyroidism in vitro and in vivo. Transfection of small interfering RNA (siRNA) against human parathyroid hormone into monolayers of parathyroid cells cultured from these patients caused a dose-dependent decrease of secretion and mRNA levels with 80% or more suppression using 40 nM siRNA. Parathyroid cells cultured on non-adherent plastic produced spheroid cell aggregates which secreted parathyroid hormone for more than 150 days. Transfection of these spheroids with 50 nM targeted siRNA decreased parathyroid hormone production to 20% of the control level, with half of them being suppressed for 50 days. When parathyroid cells were transplanted into the livers of athymic nude mice, plasma human parathyroid hormone rose to 100-300 pg/ml within one month and remained at about this level for at least 39 days. Systemic delivery of hormone-targeted siRNA into these mice caused a dose-dependent suppression of circulating human parathyroid hormone for at least one month, with a maximum 80% suppression achieved by 80 microg of siRNA. Our study shows that hormone secretion by parathyroid cells of patients with secondary hyperparathyriodism can be suppressed both in vitro and in vivo by targeted siRNAs.

Relationship between the weight of parathyroid glands and their secretion of parathyroid hormone in hemodialysis patients with secondary hyperparathyroidism.

Secondary hyperparathyroidism is one of the common and important abnormalities of mineral metabolism in hemodialysis patients. In this study we investigated the relationship between the weight of individual parathyroid glands (PTG) and their secretion of parathyroid hormone (PTH). Sixty-four PTGs in 16 patients undergoing parathyroidectomy (PTx) at our hospital were included in this study. Patients' ages ranged from 34 to 68 years (60.3 +/- 6.6 years). They were undergoing maintenance dialysis therapy for 81-256 months (175.3 +/- 56.0 months). The cause of end-stage renal failure was chronic glomerulonephritis in all patients. We measured whole PTH (wPTH) levels before PTx and 15 min after the resection of each individual gland (Delta whole PTH). A positive correlation was found between the weight of individual gland and ultrasonography (US) size of individual PTG (r = 0.91, P < 0.001, N = 53). A positive correlation was found between the total mass of the gland and the total volume of PTG on US (r = 0.896, P < 0.001, N = 16). A positive correlation was found between the mass of each individual gland and Delta whole PTH (r = 0.625, P < 0.001, N = 64); however, massive PTGs did not secrete more whole PTH per unit mass (0.01 g). Determination of the volume of PTGs by US is a good indicator of their weight. Larger PTGs secrete more whole PTH per gland, whereas these PTGs did not have the ability to secrete more PTH per unit volume.

Atrial fibrillation induced by post-parathyroidectomy transient thyrotoxicosis.

We describe a 59-year-old Japanese woman with post-parathyroidectomy transient thyrotoxicosis and atrial fibrillation. She underwent parathyroidectomy for secondary hyperparathyroidism due to chronic renal failure. Three days after surgery, she complained of palpitation and chest pain due to atrial fibrillation. Results of thyroid function tests were compatible with thyrotoxicosis. Twelve days after parathyroidectomy, the elevated level of free thyroxine decreased spontaneously to the normal range. These features were compatible with post-parathyroidectomy transient thyrotoxicosis. No further recurrences of thyrotoxicosis or atrial fibrillation were observed for one year. This is the first report of atrial fibrillation induced by post-parathyroidectomy transient thyrotoxicosis.

Indication and efficacy of PEIT in the management of secondary hyperparathyroidism.

Control of secondary hyperparathyroidism (SHPT) using active vitamin D analogues becomes difficult in advanced SHPT, because the enlarged parathyroid glands (PTGs) are resistant to medical therapy. Percutaneous ethanol injection therapy (PEIT) has been widely used in Japan since the 1990s as a surgical intervention for advanced SHPT, by selectively destroying only the enlarged glands with nodular hyperplasia (i.e. >0.5 cm(3), measured by ultrasonography). If there is only one PTG with nodular hyperplasia, PEIT will be successful with a small number of injections, and it then becomes possible to maintain target levels of parathyroid hormone by treatment with active vitamin D analogues. Recent studies have demonstrated that in the advanced phase of SHPT, it is desirable to perform PEIT when it is restricted to patients with not more than one PTG larger than 0.5 cm(3) in terms of superior prognosis can be obtained including efficacy, low recurrence, and long-term remission period.

Long-term prognosis of parathyroid function in chronic dialysis patients after PEIT-a single-centre trial.

Background. Secondary hyperparathyroidism (SHPT) is a common complication observed in long-term dialysis patients. Percutaneous ethanol injection therapy (PEIT) of parathyroid glands (PTGs) is now established in Japan as a treatment option for SHPT. In this study, to elucidate the factors influencing efficacy in 1 year and relapse following PEIT, we analysed the long-term prognosis of parathyroid function that is known to have the greatest effect on therapeutic results. Methods. The study design was a retrospective cohort study. We studied 104 patients with SHPT, who underwent PEIT at Tokai University Hospital between January 1993 and December 2002, and we followed them up until January 2008. The effective group reached intact parathyroid hormone of 200 pg/ml or less, corrected calcium (Ca) of 10.5 mg/dl or less and phosphate (P) of 6.0 mg/dl or less. The ineffective group failed to achieve these criteria. Results. Among the 104 patients, 66 patients (63%) fulfilled the criteria for the effective group within the first year of PEIT. Using the multivariate logistic regression analysis, the number of PTGs before PEIT was a significant risk factor to deviate from the criteria. At the end of the surveillance period, 31 patients (30%) fulfilled the criteria, and their SHPT was controlled with PEIT. Using the multivariate logistic regression analysis, more than three PTGs at the beginning, and the increase in PTGs during the observation period were significant risk factors to deviate from the criteria. In conclusion, superior results with PEIT are obtained in terms of efficacy, remission period and risk of relapse, regardless of the size of the gland.

Reversed whole PTH/intact PTH ratio as an indicator of marked parathyroid enlargement: five case studies and a literature review.

Parathyroid hormone (PTH) levels detected by intact PTH assays are generally higher than those detected by the whole PTH assay because the latter does not detect non-(1-84) PTH fragments, mainly PTH (7-84). Rare exceptions to this rule have been reported in patients with severe primary or secondary hyperparathyroidism and parathyroid carcinoma. Overproduction of an N-form of PTH other than PTH (1-84) has been observed in the sera of these patients. We report five additional cases with the reversed whole PTH/intact PTH ratio associated with severe hyperparathyroidism in haemodialysis patients. Three patients demonstrated enlargement of a single hypervascular gland, whereas the other two had undergone surgical parathyroidectomy and later showed recurrent hyperparathyroidism due to progressive autograft hyperplasia. In the case of a single enlarged gland, the pathological pattern and heterogeneous expression of parathyroid adenomatosis 1/cyclin D1 suggested it to be a single nodule of uraemic hyperparathyroidism rather than sporadic primary adenoma. These cases suggested that the reversed whole PTH/intact PTH ratio could be an indicator of marked parathyroid enlargement. Further studies are required to elucidate the clinical significance of the reversed whole PTH/intact PTH ratio in haemodialysis patients.

Efficacy of percutaneous ethanol injection therapy (PEIT) is related to the number of parathyroid glands in haemodialysis patients with secondary hyperparathyroidism.

BACKGROUND: Percutaneous ethanol injection therapy (PEIT) is used for advanced secondary hyperparathyroidism. We investigated the efficacy, remission period and risk of relapse to determine the effect of the number of hyperplastic glands and other factors on the therapeutic effect of PEIT. METHODS: We studied 321 patients divided into two groups: effective [serum corrected calcium (cCa) level < or =10.5 mg/dl and serum intact parathyroid hormone (iPTH) level < or =250 pg/ml], and ineffective (failed to achieve the target levels). Advanced hyperplasia was defined as an estimated volume > or =0.5 cm(3) on ultrasonography. RESULTS: PEIT was effective in 201 patients (62.6%), in whom serum iPTH levels dropped from 603+/-292 to 183+/-62 pg/ml (ng/l) and serum cCa levels from 10.7+/-0.8 to 10.1+/-0.5 mg/dl. Univariate analysis identified age, the number of hyperplastic glands and iPTH level as factors related to the efficacy of PEIT. The odds ratio for success vs failure by multivariate analysis was 0.55 times for the number of hyperplastic glands > or =0.5 cm(3) (> or =2 vs 0,1) and 0.29 times for iPTH (> or =500 vs <500 pg/ml). Using the Kaplan-Meier method, the number of hyperplastic glands > or =0.5 cm(3) (> or =2 vs 0,1) was a factor affecting the remission period, with a remission significantly longer seen in the group with one hyperplastic gland (P=0.0025). CONCLUSIONS: Superior results in efficacy rate, remission period and risk of relapse are obtained when PEIT is restricted to patients with one hyperplastic gland > or =0.5 cm(3).

Present status and perspectives of bioartificial kidneys.

Currently, hemodialysis is not adequate as a renal replacement therapy because it provides intermittent treatment and does not provide the metabolic function of renal tubules. The next generation of artificial kidney should replace intermittent hemodialysis with continuous hemofiltration and provide the full metabolic function of renal tubules. The current decade has witnessed the development of bioartificial kidneys using artificial membranes and renal tubular epithelial cells. Active transport and metabolic functions were confirmed in the confluent monolayers of tubular cells on artificial membranes. Bioartificial kidneys have succeeded in improving the prognosis of patients with multiple organ dysfunction, presumably by lowering plasma cytokine levels in patients. For successful treatment of chronic renal failure using bioartificial kidneys, it is necessary to overcome some technical hurdles such as improving the antithrombogenic properties of the surface of artificial membranes and prolonging the function of renal tubule cells on an artificial membrane. Transfection of functional protein genes into renal tubule cells enables bioartificial tubule devices to increase their transport capacity and metabolic functions such as digoxin secretion and water transport. The development of wearable roller pumps is also essential for the clinical application of a continuous treatment system.

Early hospital readmission was less likely for hemodialysis patients from facilities with longer median length of stay in the DOPPS study.

The length of hospital stay is considered to influence hospital readmission in general. The Dialysis Outcomes and Practice Patterns Study (DOPPS), an international prospective observational study undertaken to establish a relationship between facility practices and dialysis outcomes, started in 1996. Results suggest that the duration of hospital stay is significantly correlated with the probability of early readmission in dialysis patients. Thus, early hospital readmission was observed to be less likely for hemodialysis patients from facilities with longer median length of stay. The lengths of hospital stay for hemodialysis patients differed in the three continents studied. Although socioeconomic pressures may drive the lengths of hospital stay, the duration of hospitalization should be determined keeping in mind the safety of clinical course for each disease. In this forum, a 47-year-old female hemodialysis patient with severe secondary hyperparathyroidism, who had been treated with hemodialysis for 21 years, was hospitalized with severe clinical symptoms. Although the clinical symptoms disappeared 10 days after total parathyroidectomy with autotransplantation, severe hypocalcemia persisted despite large amounts of intravenous calcium gluconate. This patient was hospitalized for a long duration owing to the large calcium deficit in her body. Had the length of her hospital stay been shortened, either she could have needed rehospitalization or her condition could have worsened.

Pyridoxamine improves functional, structural, and biochemical alterations of peritoneal membranes in uremic peritoneal dialysis rats.

BACKGROUND: We previously suggested that biochemical alterations of peritoneal membrane associated with long-term peritoneal dialysis might be, at least in part, accounted for by reactive carbonyl compounds overload originating both from uremic circulation and heat sterilization of glucose peritoneal dialysis fluid. In the present study, we utilized a uremic rat model on peritoneal dialysis and evaluated the protective effects of pyridoxamine, a recently developed inhibitor of advanced glycation end product (AGE), on structural, functional, and biochemical alterations of peritoneal membrane. METHODS: Uremic rats were generated by subtotal nephrectomy, some of which were undergone peritoneal dialysis with dialysate and/or given intraperitoneal pyridoxamine. Functional [dialysate/plasma ratio (D/P)(urea, creatinine), D/D(0 glucose)], structural (density of blood vessels in peritoneal membrane tissues), and molecular biochemical [formation of pentosidine, an AGE, by high-performance liquid chromatography (HPLC) assay and expressions of vascular endothelial growth factor (VEGF), and fibroblast growth factor 2 (FGF-2), by semiquantitative polymerase chain reaction (PCR) and/or immunohistochemistry] alterations of peritoneal membrane were assessed. RESULTS: Uremic peritoneal membrane was characterized by an increased functional area of exchange for small solutes between blood and dialysate, vascular proliferation, increased AGE genesis, and up-regulated expressions of angiogenic cytokines. The peritoneal membrane alterations associated with peritoneal dialysis are similar but more severe than those in uremia without peritoneal dialysis. Pyridoxamine given in uremic rats with peritoneal dialysis significantly improved functional and structural alterations. This improvement was accompanied by reduction of AGE accumulation and of angiogenic cytokines expressions. CONCLUSION: Peritoneal carbonyl stress derived from uremia as well as peritoneal dialysis procedure might contribute to the vascular proliferation through induction of bioactive molecules and to an increased functional area, eventually leading to ultrafiltration failure. Pyridoxamine may be beneficial in protection of uremic peritoneal membrane on peritoneal dialysis.

Co-expression of parathyroid hormone and chromogranin A in secondary hyperparathyroidism: a functional marker for secretory activity of hyperplastic nodules.

The relationship between secretion of parathyroid hormone (PTH) and biologic characteristics, including cell proliferation or monoclonality, is not yet fully understood. To evaluate secretory activity of glands or nodules histopathologically, we focused on the co-expression of chromogranin A (CgA) and parathyroid hormone (PTH) in each gland or nodule. A total of 55 glands from 38 patients with normal parathyroid glands, hyperplastic glands (diffuse and nodular) and primary adenomas were compared. Co-expression of PTH and CgA was decreased to 44.4% in diffuse hyperplastic glands, and to 39.6% in 91 hyperplastic nodules, in contrast to normal glands and primary adenomas that showed constant co-expression of PTH and CgA. Immunohistochemical study of PTH showed a coarse granular pattern predominantly in PTH-positive/CgA-positive nodules, and a dot-like pattern mainly in PTH-positive/CgA-negative nodules. Laser scanning microscopy and immunoelectron microscopy confirmed that a dot-like pattern is based on a positive reaction of PTH at the Golgi apparatus. MIB-1 LI was 12.6 +/- 11.6 in PTH-positive/CgA-positive, and 19.3 +/- 27.3 in PTH-positive/CgA-negative nodules. In conclusion, a combination of PTH and CgA could provide more information about the physiologic state of secretory activity of each nodule than does the simple observation of PTH immunoreactivity.

Long-term (3 years) prognosis of parathyroid function in chronic dialysis patients after percutaneous ethanol injection therapy guided by colour Doppler ultrasonography.

BACKGROUND: Secondary hyperparathyroidism (2HPT) is one of the most important complications in chronic dialysis (CD) patients. Percutaneous ethanol injection therapy (PEIT) of the parathyroid glands was introduced initially as an alternative treatment to parathyroidectomy and, with the technical progress of parathyroid imaging, it has now become a useful adjunct to medical therapy. The present study examined the possibility of maintaining parathyroid function in the long term (3 years) after PEIT. METHOD: PEIT, guided by power-Doppler flow mapping, was performed in 33 CD patients with severe 2HPT, and all glands >5.0 mm in diameter were destroyed. RESULTS: All patients showed a decline in the serum intact parathyroid hormone (i-PTH) concentration, on average from 695.5 to 248.0 pg/ml, after 1 year. After 3 years, the i-PTH concentration was controlled at <300 pg/ml in 85% of patients. The mean serum alkaline phosphatase (ALP) concentration also decreased from 322.7 to 154.4 IU/l after 1 year. In 76% of patients, ALP was maintained within the normal range (between 76 and 260 IU/l) at 3 years. Patients were classified into four groups according to the number of parathyroid glands detected by ultrasonography: one gland in group 1, two in group 2, three in group 3, and four in group 4. At 3 years after PEIT, i-PTH was controlled at <300 pg/ml in 100, 79, 83 and 82% of the patients in groups 1-4, respectively. CONCLUSIONS: It was possible to maintain long-term parathyroid function after PEIT in patients with 2HPT by using medical therapy, such as oral calcitriol pulse therapy and additional PEIT.

Long-term prognosis of parathyroid function for chronic dialysis patients after minimally invasive radioguided parathyroidectomy (MIRP).

BACKGROUND: Minimally invasive radioguided parathyroidectomy (MIRP) for primary hyperparathyroidism for one gland, located by scanning with technetium 99m-labelled sestamibi (MIBI), has been performed. Total parathyroidectomy with autotransplantation or percutaneous ethanol injection therapy (PEIT) for severe secondary hyperparathyroidism (2HPT) has also been performed. METHODS: The present study examined the possibility of maintaining parathyroid function within a target range [intact parathyroid hormone (i-PTH)

Prognosis of parathyroid function after minimally invasive radioguided parathyroidectomy (MIRP) and percutaneous ethanol injection therapy (PEIT) for primary hyperparathyroidism.

During parathyroidectomy (PTx) for primary hyperparathyroidism (PHP), we surgically explored the contralateral parathyroid glands as well as those whose localization was clarified by ultrasonography and parathyroid scintigraphy. Although it is important to explore the contralateral side and other glands, we frequently treat only the gland whose localization is confirmed. Recently, we have performed minimally invasive radioguided parathyroidectomy (MIRP) that resects only one gland observed on the imaging under technetium 99m-labeled sestamibi (MIBI) scanning guidance after obtaining prior informed consent. In this surgery, even if recurrence is observed contralaterally, it is possible to apply a similar procedure to the contralateral side again. We examined six PHP patients who underwent MIRP and two PHP patients treated with percutaneous ethanol injection therapy (PEIT). The follow-up period was 2 years. PEIT was selected as a treatment method for two patients based on the patients' characteristics. When only one gland is treated, the efficacy of PEIT was considered to be similar to that of MIRP.