Efficacy and safety of hepatitis A vaccination in kidney transplant recipients.

BACKGROUND: In recent years, symptomatic hepatitis A virus (HAV) infection has been reported with increasing frequency in Korea. Therefore, HAV vaccination should be considered in kidney transplant recipients (KTRs). The study investigated the efficacy and safety of HAV vaccination in KTRs under modern triple immunosuppressive agents. METHODS: We evaluated the seroprevalence of anti-HAV immunoglobulin-G (IgG) in KTRs who had visited the Seoul National University Hospital from March 2011 to August 2012. Seronegative patients were immunized with 2 doses of HAV vaccine at a 6-month interval. Seroconversion of anti-HAV IgG was determined 1 month after the second vaccine dose, and adverse effects were monitored after each vaccination. RESULTS: Among a total 416 KTRs who were screened, 338 (81.2%) patients were seropositive for anti-HAV IgG. However, among patients who were under 40 years of age, only 31.8% were seropositive. Fifty-two seronegative recipients (mean age 34.1 years, 71.2% male) had received 2 doses of vaccine, and only 14 of these patients (26.9%) showed seroconversion. Vaccine responders had lower serum creatinine (1.19 ± 0.24 vs. 1.45 ± 0.49 mg/dL, P = 0.013), higher plasma hemoglobin levels (14.4 ± 1.9 vs. 12.8 ± 1.8 g/dL, P = 0.006), and had lower tacrolimus use than cyclosporine use (57.1% vs. 84.2%, P = 0.040) compared with non-responders. Responders had a tendency of taking lower dose of prednisolone (3.5 ± 1.6 vs. 4.3 ± 1.2 mg/day, P = 0.076), and having fewer infection events (14.3 vs. 40.5%, P = 0.076). Multivariate analysis indicated that higher hemoglobin levels and lower serum creatinine levels were significant prognostic factors for seroconversion. Overall, the vaccine was well tolerated in all patients. CONCLUSION: HAV IgG screening is necessary for KTRs, especially young recipients. HAV vaccination was safe in KTRs; however, poor response to HAV vaccination makes it important to identify seronegative patients as early as possible and vaccinate them before end-stage renal disease occurs.

Capillary wave dynamics of thin polymer films over submerged nanostructures.

The surface dynamics of thin molten polystyrene films supported by nanoscale periodic silicon line-space gratings were investigated with x-ray photon correlation spectroscopy. Surface dynamics over these nanostructures exhibit high directional anisotropy above certain length scales, as compared to surface dynamics over flat substrates. A cutoff length scale in the dynamics perpendicular to the grooves is observed. This marks a transition from standard over-damped capillary wave behavior to suppressed dynamics due to substrate interactions.

An attention mechanism-based LSTM network for cancer kinase activity prediction.

Despite the endeavours and achievements made in treating cancers during the past decades, resistance to available kinase drugs continues to be a major problem in cancer therapies. Thus, it is highly desirable to develop computational models that can predict the bioactivity of a compound against cancer kinases. Here, we present a Long Short-Term Memory (LSTM) framework for predicting the activities of lead molecules against seven different kinases. A total of 14,907 compounds from the ChEMBL database were selected for model building. Two different molecular representations, namely, 2D descriptors and MACCS fingerprints were subjected to the LSTM method for the training process. We also successfully integrated an attention mechanism into our model, which helped us to interpret the contribution of chemical features on kinase activity. The attention mechanism extracted the significant chemical moieties more effectively by taking them into consideration during the activity prediction. The recorded accuracies in the test sets for both 2D descriptors and MACCS fingerprints-based models were 0.81 and 0.78, respectively. The receiver operating characteristic curve (ROC)-area under the curve (AUC) score for both models was in the range of 0.8-0.99. The proposed framework can be a good starting point for the development of new cancer kinase drugs.

Sequential occurrence of life-threatening hypokalemia and rebound hyperkalemia associated with barbiturate coma therapy.

OBJECTIVE: To report a sequential occurrence of life-threatening hypokalemia and rebound hyperkalemia following barbiturate coma therapy. CASE HISTORY: A 53-year-old man was admitted to the division of nephrology due to sudden development of severe hypokalemia. The patient had been treated following a clinical diagnosis of traumatic subarachnoid hemorrhage and subdural hematoma. Barbiturate coma therapy had been performed on this patient. He developed fatal hypokalemia 10 hours after the start of thiopental administration which did not respond to potassium supplementation. The lowest potassium level following barbiturate coma therapy was 1.0 mmol/l. Severe bradycardia and cardiac arrest developed, which necessitated cardiac massage and treatment with epinephrine and atropine. He recovered from cardiac arrest. When thiopental infusion was suddenly stopped, the potassium level increased to 8.9 mmol/l, which required quick administration of calcium gluconate and infusion of glucose solution mixed with regular insulin. Despite such management, he developed asystole. After direct current cardioversion and emergency hemodialysis, he recovered from cardiac arrest and his serum potassium level was stabilized. CONCLUSION: We recommend that clinicians must be aware of the potential occurrence of severe hypokalemia, which is rare but fatal, following barbiturate coma therapy. Rebound hyperkalemia, which is fatal, may also occur following cessation of thiopental infusion. Clinicians should also be aware of this potential complication. Further studies are needed to elucidate the precise mechanism of this clinical event.

Changes in Bone Mineral Density After Kidney Transplantation.

BACKGROUND: Numerous studies have shown that osteoporosis is common in kidney transplant recipients. However, the change in bone mineral density after kidney transplantation (KT) is not fully understood. METHODS: Thirty-nine kidney transplant recipients with bone densitometry at pretransplant and 24 months after KT were reviewed. RESULTS: The recipients' median age (44.5 ± 10.7 years) and dialysis duration before KT (4.2 ± 3.4 years) were recorded. The T-scores of the lumbar spine and femur neck at 24 months after KT were positively associated with the respective pretransplant T-score (P < .001 in the lumbar spine and P < .001 in the femur neck). However, the T-score after KT did not show significant change (P = .680 in lumbar spine, P = .093 in femur neck). Changes in the T-scores of the lumbar spine and femur neck over 24 months (delta T-score) were negatively associated with the respective pretransplant T-scores (P = .001 in lumbar spine, P = .026 in femur neck). Changes in the T-scores of the lumbar spine and femur neck over 24 months (delta T-score) were also associated with the pretransplant T-scores after the adjustment of other variables. CONCLUSION: The change of bone mineral density was related with pretransplant bone mineral density. Careful follow-up of bone densitometry for KT recipients was needed.

Suspected Frequent Relapsing IgG4-related Lung Disease in Kidney Transplant Patient: A Case Report.

Besides the initial description of IgG4-related pancreatic disease, other sites are now commonly involved. However, occurrence of IgG4-related disease is rare in organ transplanted patients. A 57-year-old man who received a kidney transplantation presented with recurrent dyspnea on exertion. A computed tomography scan of the chest revealed bilateral interlobular septal thickening and multiple tubular and branching small nodular lesions in the right upper lobe, and mass-like consolidation of the left middle lobe. Despite no elevation of serum IgG4 level, a percutaneous core needle biopsy on consolidative mass showed interstitial fibrosis and infiltration of IgG4-positive plasma cells to be more than > 20 per high power field. After treatment with glucocorticoids and rituximab, the consolidative mass of the left middle lobe disappeared.

The C/EBP-binding region and adjacent sites regulate expression of the adipose P2 gene in human preadipocytes.

Human preadipocytes contain nuclear factors that specifically bind to the AE-1 sequence, previously demonstrated as an enhancer element in the regulation of adipose P2 gene expression during 3T3 adipose differentiation. By transient transfection and in vivo competition experiments, the trans-acting factors were found to bind either to the C/EBP recognition site in the AE-1 sequence and act as a negative regulator or to the adjacent site (termed 3' AE-1) and act as a positive regulator of adipose P2 gene activity in human preadipocytes.

A direct role for C/EBP and the AP-I-binding site in gene expression linked to adipocyte differentiation.

Adipocyte differentiation is accompanied by the transcriptional activation of many new genes, including the gene encoding adipocyte P2 (aP2), an intracellular lipid-binding protein. Using specific deletions and point mutations, we have shown that at least two distinct sequence elements in the aP2 promoter contribute to the expression of the chloramphenicol acetyltransferase gene in chimeric constructions transfected into adipose cells. An AP-I site at -120, shown earlier to bind Jun- and Fos-like proteins, serves as a positive regulator of chloramphenicol acetyltransferase gene expression in adipocytes but is specifically silenced by adjacent upstream sequences in preadipocytes. Sequences upstream of the AP-I site at -140 (termed AE-1) can function as an enhancer in both cell types when linked to a viral promoter but can stimulate expression only in fat cells in the intact aP2 promoter. The AE-1 sequence binds an adipocyte protein identical or very closely related to an enhancer-binding protein (C/EBP) that has been previously implicated in the regulation of several liver-specific genes. A functional role for C/EBP in the regulation of the aP2 gene is indicated by the facts that C/EBP mRNA is induced during adipocyte differentiation and the aP2 promoter is transactivated by cotransfection of a C/EBP expression vector into preadipose cells. These results indicate that sequences that bind C/EBP and the Fos-Jun complex play major roles in the expression of the aP2 gene during adipocyte differentiation and demonstrate that C/EBP can directly regulate cellular gene expression.

Effects of triethyl phosphate and nitrate on electrokinetically enhanced biodegradation of diesel in low permeability soils.

Bench-scale experiments for electrokinetically enhanced bioremediation of diesel in low permeability soils were conducted. An electrokinetic reactor (ER) was filled with kaolin that was artificially contaminated with diesel at a level of 2500 mg kg(-1). A constant voltage gradient of 1.0 V cm(-1) was applied. In phosphorus transport experiments, KH2PO4 was not distributed homogeneously along the ER, and most of the transported phosphorus was converted to water-insoluble aluminum phosphate after 12 days of electrokinetic (EK) operation. However, the advancing P front of triethyl phosphate (TEP) progressed with time and resulted in uniform P distribution. The treatments employed in the electrokinetically enhanced bioremediation of diesel were control (no addition of nitrogen and phosphorus), NP (KNO3+ KH2PO4), NT (KNO3+ TEP), UP (urea+ KH2PO4), and UT (urea+TEP). Analysis of effluent collected during the first 12 days of EK operation showed that diesel was not removed from the kaolin. After nutrient delivery, using the EK operation, the ER was transferred into an incubator for the biodegradation process. After 60 days of biodegradation, the concentrations of diesel in the kaolin for the NP, NT, UP, UT, and control treatments were 1356, 1002, 1658, 1612, and 2003 mg kg(-1), respectively. The ratio of biodegraded diesel concentration to initial concentration (2465 mg kg(-1)) in NP, NT, UP, UT, and control were 45.0%, 59.4%, 32.7%, 34.6%, and 18.7%, respectively. This result showed that TEP, treated along with NO3-, was most effective for the biodegradation of diesel. TEP was delivered more efficiently to the target zones and with less phosphorus loss than KH2PO4. However, this facilitated phosphorus delivery was effective in biodegrading diesel under anaerobic conditions only when electron acceptors, such as NO3-, were present.

Effect of Changes in Body Mass Index on Cardiovascular Outcomes in Kidney Transplant Recipients.

BACKGROUND: A higher body mass index (BMI) before kidney transplantation (KT) is associated with increased mortality and allograft loss in kidney transplant recipients (KTRs). However, the effect of changes in BMI after KT on these outcomes remains uncertain. The aim of this study was to investigate the effect of baseline BMI and changes in BMI on clinical outcomes in KTRs. METHODS: A total of 869 KTRs were enrolled from a multicenter observational cohort study from 2012 to 2015. Patients were divided into low and high BMI groups before KT based on a BMI cutoff point of 23 kg/m2. Differences in acute rejection and cardiovascular disease (CVD) between the 2 groups were analyzed. In addition, clinical outcomes across the 4 BMI groups divided by BMI change 1 year after KT were compared. Associations between BMI change and laboratory findings were also evaluated. RESULTS: Patients with a higher BMI before KT showed significantly increased CVD after KT (P = .027) compared with patients with a lower BMI. However, among the KTRs with a higher baseline BMI, only persistently higher BMI was associated with increased CVD during the follow-up period (P = .003). Patients with persistently higher BMI had significantly decreased high-density lipoprotein cholesterol and increased hemoglobin, triglyceride, and hemoglobin A1c levels. Baseline BMI and post-transplantation change in BMI were not related to acute rejection in KTRs. CONCLUSIONS: BMI in the 1st year after KT as well as baseline BMI were associated with CVD in KTRs. More careful monitoring of obese KTRs who do not undergo a reduction in BMI after KT is required.

Isolation of a novel mycovirus OMIV in Pleurotus ostreatus and its detection using a triple antibody sandwich-ELISA.

A novel mycovirus was isolated from a diseased mushroom, Pleurotus ostreatus, using a purification procedure involving polyethylene glycol (PEG)-NaCl precipitation, differential centrifugation, and equilibrium centrifugation in a CsCl gradient. The virion was a 43 nm isometric virus encapsulating double-stranded RNA (dsRNA) genome of 2.1, 2.0, 1.9, and 1.7 kbp with a coat protein (CP) of 58 kDa. The new mycovirus was named Oyster Mushroom Isometric Virus (OMIV). A triple antibody sandwich-ELISA (TAS-ELISA) system was constructed to detect OMIV in the mushroom using an anti-OMIV mouse monoclonal antibody and an anti-OMIV rabbit polyclonal serum. The TAS-ELISA system was sensitive enough to allow detection of OMIV in the mushroom with the naked eye. It detected successfully virus particles from 0.6 mg of diseased tissue as well as 0.4 microg/ml purified virus preparation.

Can Kidney Transplantation Improve Arterial Stiffness in End-Stage Renal Patients?

BACKGROUND: Arterial stiffness is associated with cardiovascular disease in end-stage renal disease (ESRD) and after kidney transplantation. We examined how kidney transplantation influences brachial-ankle pulse-wave velocity (baPWV) in ESRD patients. METHODS: The prospective observational study enrolled 67 patients who underwent successful kidney transplantation. Serial baPWV and biochemical parameters were measured before surgery and 6 months, 1 year, and 2 years after transplantation. RESULTS: baPWV prior to kidney transplantation and 6 months, 1 year, and 2 years after transplantation was 1533 ± 261 cm/s, 1417 ± 254 cm/s, 1414 ± 285 cm/s, and 1384 ± 233 cm/s, respectively. baPWV and biochemical parameters including alkaline phosphatase, intact parathyroid hormone, and 1,25 hydroxyvitamin D improved significantly at 6 months (P < .05), but there were no changes between 6 months and 2 years after transplantation. The majority of patients (73%) improved, whereas the remainder showed progression of baPWV after transplantation. Sixty-three percent of all kidney transplantation patients displayed higher baPWV than the healthy control subjects at 6 months after transplant. CONCLUSIONS: In the majority of patients, baPWV improved soon after kidney transplantation but overall remained higher than in the generally healthy population.

Sequential Changes of Vitamin D Level and Parathyroid Hormone After Kidney Transplantation.

BACKGROUND: Numerous studies have shown that vitamin D deficiency is common in end stage renal disease patients. However, change of the vitamin D deficiency after kidney transplantation is not fully understood. METHODS: Twenty-five kidney transplant (KT) recipients with serum 25-hydroxyvitamin D (25D) level, 1,25-dihydroxyvitamin D (1,25D) level, parathyroid hormone (PTH) level before and 6, 12, and 24 months after transplantation were reviewed. RESULTS: Serum PTH level and 1,25D level showed significant changes at 6 months after transplantation, but did not show further change at 12 months. 25D level did not increase within 6 months after transplantation. However, 25D level showed gradual increase after 6 months. The proportion of recipients with 25D deficiency (<10 ng/mL) was 68%, 40%, and 28% before, 12 months after, and 24 months after transplantation, respectively. Patients with vitamin D deficiency at 24 month were younger at transplantation (37.7 ± 9.6 y vs 48.2 ± 8.0 y; P = .029) and had lower 25D level before transplantation (7.96 ± 1.74 ng/mL vs 10.59 ± 4.35 ng/mL; P = .041). CONCLUSIONS: 25D deficiency is persistent in almost kidney transplant recipients even 12 months after transplantation, although serum PTH levels decrease and serum 25D levels increase after transplantation.

Health-Related Quality of Life of Kidney Transplantation Patients: Results from the KoreaN Cohort Study for Outcome in Patients With Kidney Transplantation (KNOW-KT) Study.

BACKGROUND: As patient and graft survival rates have been improving after kidney transplantation, health-related quality of life (HR-QOL) has become an important indicator of effective treatment. This study aimed to evaluate changes in HR-QOL after kidney transplantation. MATERIALS AND METHODS: The KoreaN cohort study for Outcome in patients With Kidney Transplantation (KNOW-KT) is a multicenter, observational, 9-year, cohort study. The HR-QOL of patients in the KNOW-KT study was assessed before transplantation and 2 years after transplantation using the Kidney Disease Quality of Life Short Form (KDQOL-SF) including chronic kidney disease targeted area and the Medical Outcome Study 36-item Short Form Health Survey (SF-36). Multivariate linear regression was used to identify significant factors associated with follow-up QOL scores. RESULTS: A total of 175 patients from 8 centers were analyzed. All QOL scores including the total QOL score, chronic kidney disease targeted score, and SF-36 at the 2-year follow-up were significantly increased compared to baseline values. Both physical and mental scale scores were improved after transplantation. CONCLUSION: The QOL scores for both the mental and physical scales were improved at 2 years after kidney transplantation. High glomerular filtration rate at 2 years, high baseline QOL score, and low body mass index were associated with good follow-up QOL scores. Kidney transplantation for an Asian population with end-stage renal disease can result in better QOL as well as better patient and graft survival.

Thyrotropin induces SOCS-1 (suppressor of cytokine signaling-1) and SOCS-3 in FRTL-5 thyroid cells.

TSH has multiple physiological roles: it is required for growth, differentiation, and function of the thyroid gland, and it regulates transcription of interferon-gamma (IFN-gamma)-responsive genes in thyrocytes, including genes for the major histocompatibility complex and intercellular adhesion molecule-1. This report demonstrates that TSH induces the expression of suppressor of cytokine signaling (SOCS)-1 and -3 proteins and alters the phosphorylation state of signal transducer and activator of transcription (STAT) proteins STAT1 and STAT3. The expression of SOCS-1 and SOCS-3 and the phosphorylation state of STAT1 and STAT3 were examined after treatment with TSH or IFN-gamma in either TSH-sensitive FRTL-5 thyroid cells or TSH-insensitive FRT and buffalo rat liver (BRL) cells, which lack functional TSH receptors. SOCS-1 and SOCS-3 are constitutively expressed in FRTL-5 cells, but not in FRT and BRL cells. IFN-gamma up-regulated SOCS-1 and SOCS-3 RNA and protein in FRTL-5 cells, as reported previously for nonthyroid cells. Interestingly, TSH also significantly induced SOCS-1 and SOCS-3 in FRTL-5 cells, but not in FRT and BRL cells. When SOCS-1 or SOCS-3 was overexpressed in FRTL-5 cells, STAT1 phosphorylation at Y701 and STAT1/DNA complex formation in response to IFN-gamma were reduced. Furthermore, overexpression of either SOCS-1 or SOCS-3 significantly inhibited the IFN-gamma-mediated transactivation of the rat ICAM-1 (intercellular adhesion molecule-1) promoter. TSH and IFN-gamma had different effects on STAT1 and STAT3 phosphorylation. The phosphorylation of Y701 in STAT1, which is responsible for homodimer formation, nuclear translocation, and DNA binding, was specifically stimulated by IFN-gamma, but not by TSH or forskolin. However, the phosphorylation of S727 in STAT1 was induced by IFN-gamma, TSH, and forskolin. TSH induced phosphorylation of both Y705 and S727 in STAT3, while IFN-gamma phosphorylated only the Y705. In addition, we found that SOCS-3 was associated with JAK1 and JAK2 and that these associations were stimulated by TSH. These findings demonstrate that TSH induces SOCS in thyroid cells and provides the evidence of signal cross-talk between TSH and cytokines in thyroid cells.

Involvement of JAK/STAT (Janus kinase/signal transducer and activator of transcription) in the thyrotropin signaling pathway.

TSH is an important physiological regulator of growth and function in thyroid gland. The mechanism of action of TSH depends on interaction with its receptor coupled to heterotrimeric G proteins. We show here that TSH induces the phosphorylation of tyrosine in the intracellular kinases Janus kinase 1 (JAK1) and -2 (JAK2) in rat thyroid cells and in Chinese hamster ovary (CHO) cells transfected with human TSH receptor (TSHR). The JAK family substrates STAT3 (signal transducers and activators of transcription) are rapidly tyrosine phosphorylated in response to TSH. We also find that JAK1, JAK2, and STAT3 coprecipitate with the TSHR, indicating that the TSHR may be able to signal through the intracellular phosphorylation pathway used by the JAK-STAT cascade. TSH increases STAT3-mediated promoter activity and also induces endogenous SOCS-1 (suppressor of cytokine signaling-1) gene expression, a known target gene of STAT3. The expression of a dominant negative form of STAT3 completely inhibited TSH-mediated SOCS-1 expression. These findings suggest that the TSHR is able to signal through JAK/STAT3 pathways.

Acidosis inhibits 1,25-(OH)2D3 but not cAMP production in response to parathyroid hormone in the rat.

Parathyroid hormone (PTH) is a major activator of renal proximal tubule 25-hydroxyvitamin D3-1-hydroxylase (1-OHase). Chronic metabolic acidosis (CMA) inhibits 1-OHase and reduces circulating 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] levels in rats fed a low-Ca diet (LCD, 0.002% Ca). To examine the cellular mechanism whereby CMA inhibits 1-OHase, PTH-dependent renal 1-OHase activity and cAMP were measured in proximal tubules isolated from rats fed LCD for 14 days and made acidotic by the addition of 1.5% ammonium chloride to the drinking water. Serum 1,25-(OH)2D3 and proximal tubule 1-OHase activity and cAMP content were lower in acidotic rats. hPTH-(1-34) (10(-7) M) in vitro increased cAMP content to equivalent concentrations in tubules from rats with CMA and from nonacidotic controls; however, PTH increased 1-OHase activity only in tubules from nonacidotic animals. Although forskolin increased tubule cAMP content to equivalent levels in tubules from acidotic and nonacidotic rats, 1-OHase activity declined in tubules from nonacidotic rats and remained suppressed in acidotic tubules. The results suggest that chronic metabolic acidosis inhibits the PTH activation of 1-OHase through alteration of one or more steps in a cAMP-independent messenger system. PTH and forskolin can increase cAMP production by acidotic and nonacidotic proximal tubules; however, 1-OHase activity is not restored to normal in acidotic tubules and nonacidotic tubule 1-OHase may be inhibited.

Loss of parathyroid hormone-stimulated 1,25-dihydroxyvitamin D3 production in aging does not involve protein kinase A or C pathways.

Intestinal calcium absorption declines with aging as a result of decreased renal 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] biosynthesis. At least part of the decline in 1,25-(OH)2D3 may be due to acquired resistance to parathyroid hormone (PTH) stimulation of renal 25-hydroxyvitamin D1-hydroxylase (1-OHase) activity. To test whether aging rats can increase 1,25-(OH)2D3 production in response to PTH, male rats of the same litter were fed a normal Ca diet and were sacrificed at 175-225 g (young rats) or 3 months later at 350-425 g (aging rats). At sacrifice, basal serum 1,25-(OH)2D3 levels (88 +/- 16 versus 49 +/- 8 pg/ml, P < 0.05) and in vitro renal proximal tubule 1-OHase activity (178 +/- 15 versus 77 +/- 5 pmol/mg protein/5 minutes, n = 6, P < 0.001) were lower in aging animals. rPTH-(1-34) (10(-11) or 10(-7) M) increased in vitro 1,25-(OH)2D3 secretion by perifused renal proximal tubules from young but not aging rats. For young and aging rats, rPTH-(1-34) (10(-7) M) increased proximal tubule cAMP-dependent protein kinase (PKA) activity, and lower concentrations (10(-11) M) stimulated translocation of protein kinase C (PKC) activity from cytosolic to soluble membrane proximal tubule cell fractions. The results of this study show that PTH activation of 1,25-(OH)2D3 production may involve both signaling pathways, with the PKC pathway responsive to lower concentrations of the hormone. The acquired resistance to PTH stimulation of 1,25-(OH)2D3 production in aging appears not to involve the hormonal activation of PKA or PKC.

Evidence that activation of protein kinase-C can stimulate 1,25-dihydroxyvitamin D3 secretion by rat proximal tubules.

PTH stimulates mammalian renal proximal tubule cell synthesis and secretion of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] by a Ca-dependent process. In the present study regulation of 1,25-(OH)2D3 secretion by PTH, phorbol ester 12-O-tetradecanoylphorbol 13-acetate, the Ca ionophore A23187, and calcitonin was evaluated in perifused rat proximal tubule cells isolated by collagenase digestion and centrifugation through Percoll. Tubules from rats fed a low Ca diet secreted 1,25-(OH)2D3 at a rate 2.5 times that of tubule cells from rats fed a normal Ca diet. Perifusion of tubules with human PTH-(1-34) (10(-7) M) induced an immediate and sustained increase in 1,25-(OH)2D3 secretion. Perifusion with either A23187 or 12-O-tetradecanoylphorbol 13-acetate caused transient increases in hormone secretion, while both agents perifused simultaneously resulted in a sustained increase in 1,25-(OH)2D3 secretion. Perifusion of tubule cells with the protein kinase-C (PKC) inhibitor staurosporine blocked the PTH-induced increase in 1,25-(OH)2D3 secretion. Calcitonin had no effect on 1,25-(OH)2D3 secretion rates. The results of the present studies show that an activator of PKC increases 1,25-(OH)2D3 secretion by mammalian proximal tubule cells and suggest that the phospholipase-C/PKC signalling system may mediate PTH stimulation of 1,25-(OH)2D3 secretion.