Diverse molecular causes of unsolved autosomal dominant tubulointerstitial kidney diseases.

Autosomal Dominant Tubulointerstitial Kidney Disease (ADTKD) is caused by mutations in one of at least five genes and leads to kidney failure usually in mid adulthood. Throughout the literature, variable numbers of families have been reported, where no mutation can be found and therefore termed ADTKD-not otherwise specified. Here, we aim to clarify the genetic cause of their diseases in our ADTKD registry. Sequencing for all known ADTKD genes was performed, followed by SNaPshot minisequencing for the dupC (an additional cytosine within a stretch of seven cytosines) mutation of MUC1. A virtual panel containing 560 genes reported in the context of kidney disease (nephrome) and exome sequencing were then analyzed sequentially. Variants were validated and tested for segregation. In 29 of the 45 registry families, mutations in known ADTKD genes were found, mostly in MUC1. Sixteen families could then be termed ADTKD-not otherwise specified, of which nine showed diagnostic variants in the nephrome (four in COL4A5, two in INF2 and one each in COL4A4, PAX2, SALL1 and PKD2). In the other seven families, exome sequencing analysis yielded potential disease associated variants in novel candidate genes for ADTKD; evaluated by database analyses and genome-wide association studies. For the great majority of our ADTKD registry we were able to reach a molecular genetic diagnosis. However, a small number of families are indeed affected by diseases classically described as a glomerular entity. Thus, incomplete clinical phenotyping and atypical clinical presentation may have led to the classification of ADTKD. The identified novel candidate genes by exome sequencing will require further functional validation.

Using Sensor Graphs for Monitoring the Effect on the Performance of the OTAGO Exercise Program in Older Adults.

The OTAGO exercise program is effective in decreasing the risk for falls of older adults. This research investigated if there is an indication that the OTAGO exercise program has a positive effect on the capacity and as well as on the performance in mobility. We used the data of the 10-months observational OTAGO pilot study with 15 (m = 1, f = 14) (pre-)frail participants aged 84.60 y (SD: 5.57 y). Motion sensors were installed in the flats of the participants and used to monitor their activity as a surrogate variable for performance. We derived a weighted directed multigraph from the physical sensor network, subtracted the weights of one day from a baseline, and used the difference in percent to quantify the change in performance. Least squares was used to compute the overall progress of the intervention (n = 9) and the control group (n = 6). In accordance with previous studies, we found indication for a positive effect of the OTAGO program on the capacity in both groups. Moreover, we found indication that the OTAGO program reduces the decline in performance of older adults in daily living. However, it is too early to conclude causalities from our findings because the data was collected during a pilot study.

Spectral integration of infrasound at threshold.

The present study investigated the effect of the number of spectral components on the threshold in quiet of infrasound stimuli. Stimuli with one, two, or three sinusoidal components were presented monaurally to the ear with a low-distortion infrasound reproduction system. All components of the complex tones had the same level relative to their respective pure-tone threshold. The data are consistent with the assumption of a (perceptually weighted) intensity integration at threshold: The level at threshold for each component is 3 dB lower when two components were presented simultaneously. Thresholds decrease further, when a three-tone complex is used.

Analyzing the Importance of Sensors for Mode of Transportation Classification.

The broad availability of smartphones and Inertial Measurement Units in particular brings them into the focus of recent research. Inertial Measurement Unit data is used for a variety of tasks. One important task is the classification of the mode of transportation. In the first step, we present a deep-learning-based algorithm that combines long-short-term-memory (LSTM) layer and convolutional layer to classify eight different modes of transportation on the Sussex-Huawei Locomotion-Transportation (SHL) dataset. The inputs of our model are the accelerometer, gyroscope, linear acceleration, magnetometer, gravity and pressure values as well as the orientation information. In the second step, we analyze the contribution of each sensor modality to the classification score and to the different modes of transportation. For this analysis, we subtract the baseline confusion matrix from a confusion matrix of a network trained with a left-out sensor modality (difference confusion matrix) and we visualize the low-level features from the LSTM layers. This approach provides useful insights into the properties of the deep-learning algorithm and indicates the presence of redundant sensor modalities.

Performance of a novel high sensitivity cardiac troponin I assay in asymptomatic hemodialysis patients - evidence for sex-specific differences.

Background High sensitivity assays for the determination of cardiac troponin I (cTnI) are able to reliably measure cTnI far below the 99th percentile of healthy persons (hs-cTnI) and display sex-specific differences. There is uncertainty regarding the clinical utility of hs-cTnI in asymptomatic hemodialysis (HD) patients and if sex-specific differences also apply in this cohort. Methods In this multicenter study we measured hs-cTnI and sensitive cTnI (s-TnI) concentrations (both on Siemens Centaur) in 215 HD patients from a predialytic sample to determine the prevalence of elevated concentrations above the 99th percentile, the association with baseline characteristics, prognostic accuracy for death, and sex-specific differences. Results Hs-cTnI and s-cTnI concentrations were below the 99th percentile in 93% and 85% of patients with a median concentration of 12 ng/L (interquartile range 7-66) and 19 ng/L (12; 31, p < 0.0001). Hs-cTnI and s-cTnI concentrations were independently associated with age (p < 0.05) and ischemic cardiac disease (p < 0.05), but not with residual renal function. Both hs-cTnI and s-cTnI were predictors of death after median follow-up of 2.6 years with an AUC of 0.733 and 0.744, respectively (both p < 0.0001). Important sex-differences emerged for hs-cTnI, but not for s-cTnI: first, women had significantly lower hs-cTnI concentrations than men (p = 0.03); second, hs-cTnI had significantly higher prognostic accuracy for death in women than for men (AUC 0.824 vs. 0.674, p = 0.04). Conclusions The majority of HD patients have (h)s-cTnI concentrations below the 99th percentile. High normal values are predictive of death. Hs-cTnI allows to elucidate important sex-differences in HD patients with lower concentrations and higher prognostic accuracy in women.

Systemic haemodynamics in haemodialysis: intradialytic changes and prognostic significance.

Background: Although haemodialysis (HD) leads to alterations of systemic haemodynamics that can be monitored using dilution methods, there is a lack of data on the diagnostic and prognostic significance of haemodynamic monitoring during routine HD. Methods: In this multicentre study, we measured cardiac index (CI), access flow (AF) and central blood volume index (CBVI) during a single HD session in stable HD patients (n = 215) using the Transonic HD03 monitor (Transonic, Ithaca, NY, USA). Systemic CI (SCI) was defined as CI corrected for AF. In a subset of patients (n = 82), total end-diastolic volume index (TEDVI) and total ejection fraction (TEF) were derived from dilution curves. Data were correlated with clinical parameters, cardiac biomarkers and bioimpedance measurements (body composition monitor; Fresenius Medical Care, Homburg, Germany). Mortality was assessed prospectively after a median follow-up of 2.6 years. Results: Median CI, CBVI and AF were 2.8 L/min/m2 (interquartile range 2.4-3.4), 15 mL/kg (14.5-15.7) and 980 mL/min (740-1415), respectively, at the beginning of HD. At the end of HD, CI, CBVI and AF significantly fell by -10% (-22 to 3, P < 0.0001), -9% (-23 to 3, P < 0.0001) and -4% (-13 to 5, P = 0.0004), respectively. Peripheral resistance (PR) increased slightly (P = 0.01) and blood pressure fell by -6/-3 mmHg to 128/63 mmHg (P < 0.0001). Independent predictors of ΔCI were age and ultrafiltration rate, whereas AF, overhydration and PR were protective. TEF was strongly associated with mortality [area under the dilution curve 0.77, P < 0.0001], followed by TEDVI (0.72, P = 0.0002) and SCI (0.60, P = 0.02). Conclusions: HD leads to a reduction of CI due to ultrafiltration. Haemodynamic monitoring identifies a significant number of HD patients with cardiac impairment that are at risk for increased mortality.

Sclerostin quo vadis? - is this a useful long-term mortality parameter in prevalent hemodialysis patients?

BACKGROUND/AIMS: Cardiovascular calcification contributes to the increased mortality in hemodialysis patients. Sclerostin was identified as an antianabolic bone factor causing soft tissue calcification. Data on prospective large-scale studies associating sclerostin with mortality in hemodialysis patients are so far inconsistent. METHODS: In our multicenter prospective longitudinal study following hemodialysis patients, we assessed the associations of sclerostin and bone remodeling markers with long-term mortality. We evaluated the relationship between circulating sclerostin, Fibroblast growth factor 23 (FGF23) and traditional bone remodeling markers. Sclerostin levels in hemodialysis patients were compared with healthy controls. RESULTS: We enrolled 239 hemodialysis patients with a median follow up of 1461 days. In Cox regression analysis, FGF23 (HR 1.40;95%CI 1.11-1.76), parathyroid hormone (PTH) (HR 1.80;95%CI 1.44-2.26) and alkaline phosphatase (AP) (HR 1.50;95%CI 1.10-2.04) per SD, 25(OH)vitamin D (HR 0.42;95%CI 0.23-0.76) per natural log but not sclerostin (HR 1.02;95%CI 0.75-1.38) per SD increase were associated with mortality. FGF23, PTH and AP were negatively associated with sclerostin. Among control and hemodialysis females, sclerostin levels were lower than in men. CONCLUSION: Higher FGF23, PTH, AP and lower 25(OH)vitamin D but not sclerostin predict long-term mortality. Sclerostin was negatively associated with FGF23, PTH and AP and lower in females than in males.

Mortality prediction using modern peptide biomarkers in hemodialysis patients--a comparative analysis.

BACKGROUND/AIMS: Determination of peptide biomarkers such as troponins, natriuretic peptides or the recently reported FGF23 can be useful to identify hemodialysis patients with a high risk of mortality. However, it is desirable to focus on few robust parameters to warrant their routine application. METHODS: In a prospective cohort study with 239 prevalent hemodialysis patients we studied the prognostic significance of 10 simultaneously determined modern peptide biomarkers (high sensitive troponin I and T, NT-pro-BNP, BNP, MR-pro-ANP, MR-pro-ADM, CT-pro-ET1, copeptin, FGF23 and a-Klotho) and compared them with parameters traditionally associated with mortality (PTH, Ca, Pi, albumin, CRP, cholesterol, AP). RESULTS: After a follow-up of 4 years, plasma concentration of troponins, natriuretic peptides, MR-pro-ADM, FGF23 as well as PTH, CRP, AP were significantly higher in deceased patients (n=95). Hazard ratios from cox regression on a continuous scale (doubling of plasma concentration) or relative in tertiles were highest for high sensitive troponins, followed by natriuretic peptides and MR-pro-ADM (1.6-2.0 and 2.3-5.5, resp.). C-indices were also highest for troponins (0.708-0.746), followed by natriuretic peptides (0.706-0.731). Traditional parameters had low c-indices (0.598-0.655). Stepwise cox regression revealed that among all parameters troponin I, NT-pro-BNP, PTH and CRP remained independent predictors of mortality and a composite score had the highest c-index (0.799 [0.740-0.849]). CONCLUSIONS: Among peptide biomarkers high sensitive troponins and to a lesser extent natriuretic peptides are strong predictors of mortality in asymptomatic hemodialysis patients, followed by markers of mineral-bone disease and inflammation.

How to define thresholds for level and interaural-level-difference discrimination: Insights from scedasticities and distributions.

Sensitivity to changes in the stimulus level at one or at both ears and to changes in the interaural level difference (ILD) between the two ears has been studied widely. Several different definitions of threshold and, for one of them, two different ways of averaging single-listener thresholds have been used (i.e., arithmetically and geometrically), but it is unclear which definition and which way of averaging is most suitable. Here, we addressed this issue by examining which of the differently defined thresholds yielded the highest degree of homoscedasticity (homogeneity of the variance). We also examined how closely the differently defined thresholds followed the normal distribution. We measured thresholds from a large number of human listeners as a function of stimulus duration in six experimental conditions, using an adaptive two-alternative forced-choice paradigm. Thresholds defined as the logarithm of the ratio of the intensities or amplitudes of the target and the reference stimulus (i.e., as the difference in their levels or ILDs; the most commonly used definition) were clearly heteroscedastic. Log-transformation of these latter thresholds, as sometimes performed, did not result in homoscedasticity. Thresholds defined as the logarithm of the Weber fraction for stimulus intensity and thresholds defined as the logarithm of the Weber fraction for stimulus amplitude (the most rarely used definition) were consistent with homoscedasticity, but the latter were closer to the ideal case. Thresholds defined as the logarithm of the Weber fraction for stimulus amplitude also followed the normal distribution most closely. The discrimination thresholds should therefore be expressed as the logarithm of the Weber fraction for stimulus amplitude and be averaged arithmetically across listeners. Other implications are discussed, and the obtained differences between the thresholds in different conditions are compared to the literature.

Temporal integration of infrasound at threshold.

Infrasounds are signals with frequencies below the classical audio-frequency range, i.e., below 20 Hz. Several previous studies have shown that infrasound is audible as well, provided that the sound level is high enough. Hence, the sound pressure levels at threshold are much higher than those in the classical audio-frequency range. The present study investigates how the duration and the shape of the temporal envelope affect thresholds of infrasound stimuli in quiet. Two envelope types were considered: one where the duration of the steady state was varied (plateau bursts) and one where the number of consecutive onset-offset bursts was varied (multiple bursts). Stimuli were presented monaurally to human listeners by means of a low-distortion sound reproduction system. For both envelope types, thresholds decrease with increasing duration, a phenomenon often referred to as temporal integration. At the same duration, thresholds for plateau-burst stimuli are typically lower than those for multiple-burst stimuli. The data are well described by a slightly modified version of a model that was previously developed to account for temporal integration in the classical audio-frequency range. The results suggest similar mechanisms underlying the detection of stimuli with frequencies in the infrasound and in the classical audio-frequency range. Since the model accounts for the effect of duration and, more generally, the shape of the envelope, it can be used to enhance the comparability of existing and future datasets of thresholds for infrasounds with different temporal stimulus parameters.

A system for monitoring the functional status of older adults in daily life.

Functional decline in older adults can lead to an increased need of assistance or even moving to a nursing home. Utilising home automation, power and wearable sensors, our system continuously keeps track of the functional status of older adults through monitoring their daily life and allows health care professionals to create individualised rehabilitation programmes based on the changes in the older adult's functional capacity and performance in daily life. The system uses the taxonomy of the International Classification of Functioning, Disability and Health (ICF) by the World Health Organization (WHO). It links sensor data to five ICF items from three ICF categories and measures their change over time. We collected data from 20 (pre-)frail older adults (aged [Formula: see text] 75 years) during a 10-month observational randomised pilot intervention study. The system successfully passed the first pre-clinical validation step on the real-world data of the OTAGO study. Furthermore, an initial test with a medical professional showed that the system is intuitive and can be used to design personalised rehabilitation measures. Since this research is in an early stage further clinical studies are needed to fully validate the system.

Comparison of the diagnostic performance of three natriuretic peptides in hemodialysis patients: which is the appropriate biomarker?.

BACKGROUND: Plasma concentrations of natriuretic peptides are often elevated in chronic hemodialysis patients and difficult to interpret due to accumulation, high incidence of cardiac disease and changes in volume status. Mid-regional pro-ANP is a newly developed assay whereas BNP and its fragment NT-pro-BNP are available for a longer time. In this cross-sectional study, we compared the plasma concentration of MR-pro-ANP, BNP and NT-pro-BNP in stable ambulatory hemodialysis patients (n = 239) and investigated their associations with clinical factors such as residual diuresis, cardiac status and interdialytic weight gain and with mortality. METHODS AND RESULTS: In all patients enrolled, the plasma concentration of all natriuretic peptides were largely elevated with a median concentration of 337 pg/ml (interquartile range 146-684) for BNP, 4435 pg/ml (1687-16228) for NT-proBNP and 907 pmol/L (650-1298) for MR-pro-ANP. Plasma concentration of all natriuretic peptides correlated independently with age, degree of systolic dysfunction and negatively with residual diuresis. Dependency on residual renal clearance was strongest for the fragments MR-pro-ANP and NT-pro-BNP. The plasma concentration of all natriuretic peptides was associated with mortality within 2 years of follow-up. Receiver-operated curves revealed a low sensitivity (32-45%), but high specificity for all natriuretic peptides (85-93%) resulting in a high negative predictive (82-87%). Best cut-off values obtained from were 18 611 pg/ml for NT-pro-BNP, 958 pg/ml for BNP and 1684 pmol/L for MR-pro-ANP. CONCLUSIONS: In hemodialysis patients, the fragments NTproBNP and MR-pro-ANP are largely elevated compared to BNP which is explained by accumulation. The prognostic performance of MR-pro-ANP is similar to that of NT-pro-BNP or BNP.

ECM remodelling components regulated during jaw periosteal cell osteogenesis.

Human JPCs (jaw periosteal cells) are a promising source for the engineering of cell-based osteoinductive grafts in oral surgery. For this purpose, cell characteristics of this stem cell source should be elucidated in detail. Analysis of gene expression profiles may help us to evaluate key factors and cellular targets of JPC osteogenesis. Because little is known about the interplay of osteogenic-related components, we analysed the expression of different collagen types reflecting important players for extracellular matrix assembly and of TIMPs (tissue inhibitors of metalloproteinases) responsible for the inhibition of matrix degradation. Gene expression analyses using microarrays and quantitative RT-PCR (reverse transcription-PCR) during JPC osteogenesis revealed the induction of several collagen types' expression (VII, VIII, XI and XII), and some of them (types I, VIII and XI) seemed to be susceptible to BMP-2 (bone morphogenetic protein-2) that is known to be a potent osteogenic inducer of periosteal cells. Among the TIMPs, only TIMP-4 and RECK (reversion-inducing cysteine-rich protein with Kazal motifs) expressions were strongly up-regulated during JPC osteogenesis. Proteome profiler analysis of supernatants from untreated and differentiated JPCs confirmed the gene expression data in terms of TIMP expression. In summary, we identified new collagen types and TIMPs that seem to play important roles during the osteogenesis of jaw periosteal progenitor cells.

A Deep Learning Approach for TUG and SPPB Score Prediction of (Pre-) Frail Older Adults on Real-Life IMU Data.

Since older adults are prone to functional decline, using Inertial-Measurement-Units (IMU) for mobility assessment score prediction gives valuable information to physicians to diagnose changes in mobility and physical performance at an early stage and increases the chances of rehabilitation. This research introduces an approach for predicting the score of the Timed Up & Go test and Short-Physical-Performance-Battery assessment using IMU data and deep neural networks. The approach is validated on real-world data of a cohort of 20 frail or (pre-) frail older adults of an average of 84.7 years. The deep neural networks achieve an accuracy of about 95% for both tests for participants known by the network.

Detecting Impending Malnutrition of (Pre-) Frail Older Adults in Domestic Smart Home Environments.

Malnutrition is a well-known risk factor for deteriorated physical function, disability and loss in independence in older adults. An unintended loss in body weight of more than 5% in 3 months is one indicator for malnutrition. In this study we examined the relationship between meal preparation time, hand grip strength, and body weight in order to map impending nutritional problems using ambient sensors. Data were collected in the domestic environments of 20 (pre-) frail older adults aged 85.75 y (Standard Deviation: 5.19 y) over 10-months of observation. Collecting included physical function and nutritional status of the participants and meal preparation time by a combination of motion and power sensor events. Analysis was done by rank correlation of hand grip strength, body weight, and meal preparation time. Ten participants aged 85.1 years (Standard Deviation: 4.6 y) were included. The results show a significant correlation (≥0.99) of the meal preparation time with the hand grip strength. This result validated the meal preparation time as a suitable measure for analysing the correlation between meal preparation time and body weight, and a significant correlation (≥0.99) found. Hence the meal preparation time could be used as an indicator for malnutrition. However, causalities have to be conducted by further clinical studies.

MSCA-1/TNAP selection of human jaw periosteal cells improves their mineralization capacity.

Human jaw periosteum-derived cells (JPCs) represent an alternative cell source to bone marrow-derived mesenchymal stem cells for tissue engineering applications in the oral and maxillofacial surgery. In this study we investigated how far the presence or expression of human mesenchymal stem cell antigen-1/tissue non-specific alkaline phosphatase (MSCA-1/TNAP) and LNGFR (CD271) can be utilized to select and enrich the osteogenic progenitor cell fraction from the entire JPC population. Depending on their mineralization capacity, we classified the human isolated JPCs into mineralizing (mJPCs) and non-mineralizing JPCs (nmJPCs). Flow cytometric analyses revealed that undifferentiated mJPCs expressed MSCA-1/TNAP at significant higher levels than nmJPCs at day 5 and 10 of osteogenesis. Western blot analyses showed increased MSCA-1/TNAP expression levels in mJPCs during osteogenesis, whereas in nmJPCs MSCA-1/TNAP expression remained undetectable. Using the MSCA-1 and LNGFR specific antibodies, we separated the positive and negative fractions from the entire mJPC population. In order to analyse the mineralization capacity of the MSCA-1(+) and LNGFR(+) cell subsets, we quantified the calcium deposition in both subpopulations in comparison to the respective negative subpopulations. The MSCA-1(+)/TNAP(+) cell fraction showed a significant higher osteogenic capacity compared to the MSCA-1-/TNAP- cell fraction whereas the LNGFR(+/-) cell fractions did not differ in their osteogenic potential. Our findings suggest that MSCA-1 may represent a promising osteogenic marker for mJPC.

LNGFR induction during osteogenesis of human jaw periosteum-derived cells.

Isolated jaw periosteum-derived cells (JPCs) comprise a morphologically heterogeneous population. There are no known specific surface markers that are able to distinguish between progenitors and cells of other tissue types. The aim of our study was to identify differentiation markers as predictors of JPC mineralization capacity. JPCs underwent osteogenic differentiation after cultivation in osteogenic medium containing known activators. By FACS analysis, we found the low affinity nerve growth factor receptor (LNGFR-CD271) to be induced during the first five days of osteogenesis and that it was expressed at higher levels in mineralizing JPCs (mJPCs) in comparison to non-mineralizing JPCs (nmJPCs). Similar results were obtained by semi-quantitative immunohistochemical stainings and western blot analyses. Quantitative real-time PCR results showed significantly higher LNGFR and alkaline phosphatase transcript levels in mJPCs compared to nmJPCs. LNGFR is a differentiation marker that distinguishes between mineralizing JPCs and non-mineralizing JPCs during the first phase of osteogenesis and can therefore be considered an early surface marker of osteogenic capacity in vitro.

APC sensitive gastric acid secretion.

Adenomatous polyposis coli (APC) is a tumor suppressor gene inactivated in familial adenomatous polyposis and sporadic colorectal cancer. Mice carrying a loss-of-function mutation in the apc gene (apc(Min/+)) spontaneously develop gastrointestinal tumors. APC fosters degradation of beta-catenin, which in turn upregulates the serum- and glucocorticoid-inducible kinase SGK1. SGK1 stimulates KCNQ1, which is required for luminal K+ recycling and thus for gastric acid secretion. BCECF-fluorescence was utilized to determine gastric acid secretion in isolated gastric glands from apc(Min/+) mice and their wild type littermates (apc(+/+)). Western blotting was employed to analyse beta-catenin and SGK1 expression and immunohistochemistry to determine KCNQ1 protein abundance. beta-catenin and SGK1 expression were enhanced in apc(Min/+) mice. Cytosolic pH was similar in apc(Min/+) mice and apc(+/+) mice. Na+-independent pH recovery following an ammonium pulse (DeltapH/min), which reflects H+/K+ ATPase activity, was, however, significantly faster in apc(Min/+) mice than in apc(+/+)mice. In both genotypes DeltapH/min was abolished in the presence of H+/K+ ATPase inhibitor omeprazole (100 microM). Treatment of apc(Min/+) and apc(+/+)mice with 5 microM forskolin 15 minutes prior to the experiment or increase in local K+-concentrations to 35 mM (replacing Na+/NMDG) significantly increased DeltapH/min and abrogated the differences between genotypes. The increase of DeltapH/min in apc(Min/+)mice required SGK1, as it was abolished by additional knockout of SGK1 (apc(Min/+)/sgk1(-/-)). In conclusion, basal gastric acid secretion is significantly enhanced in apc(Min/+)mice, pointing to a role of APC in the regulation of gastric acid secretion. The effect of APC requires H+/K+ ATPase activity and is at least partially due to SGK1-dependent upregulation of KCNQ1.

Acute effects of haemodialysis on pro-/anti- apoptotic genes in peripheral blood leukocytes.

Several studies have implicated a remarkable dysfunctional apoptotic state and/or response in ESRD patients. Previously published studies are controversial with respect to acute effects of haemodialysis (HD) treatment on up- or downregulation of apoptotic genes. Twenty-eight chronic HD patients were haemodialysed for 4 hours with a 4008 dialyser using high-flux membranes. For subgroup analysis, patients were separated into a low (up to 0.5 mg/dl) and a high (0.5 to 5.0 mg/dl) CRP group. Blood was drawn prior to HD and 240 min after initiation of HD. Acute changes of transcript levels encoding pro- or anti-apoptotic genes were analyzed in RNA immediately isolated from blood leukocytes using quantitative real-time PCR. In the present study, we detected a significant elevation of the death receptor CD95/Fas (induction factor (IF) 1.55 +/- 0.16), the death receptor 5 (DR5) (IF 1.17 +/- 0.08), and caspase 8 (IF 1.37 +/- 0.14) gene expression during HD. mRNA levels of the respective ligands (CD95L, TRAIL), of the caspase 5 and anti-apoptotic Bcl-2 family members such as Bcl-2 and Bcl2l2 were slightly, but not significantly, increased after HD treatment. An additional anti-apoptotic molecule, BAG3, was found to be slightly, but significantly, induced after HD (IF 1.16 +/- 0.07). In addition to being an activator of immune cells, CD40L has been shown to be strongly induced after HD treatment (IF 1.70 +/- 0.20). Subgroup analysis revealed no significant differences between low vs. high CRP patient groups or diabetic vs. non-diabetic patients. These results indicate a marked influence of routine haemodialysis treatment on the transcription of pro- and anti-apoptotic molecules and the involvement of the extrinsic pathway for apoptosis through the activation of death receptors and the initiator caspase 8. Furthermore, following dialysis, lymphocytes seem to be activated by CD40L, which represents an early T-cell activation marker.

DOCA and TGF-beta induce early growth response gene-1 (Egr-1) expression.

Renal fibrosis is characterized by excessive accumulation of extracellular matrix proteins. Recent findings show that transforming growth factor-beta (TGF-beta) induces a rapid but transient expression of early growth response gene-1 (Egr-1) by skin fibroblasts. The present study aims to define the role of Egr-1 in mineralocorticoid-induced renal fibrosis. Therefore, we transiently transfected immortalized human renal fibroblasts (TK188) with recombinant Egr-1 and analysed the transcription of several pro-fibrotic genes (Coll1A1, Coll1A2, osteopontin, TIMP-1, and CTGF). We also examined Egr-1 expression and the regulation of pro-fibrotic genes in DOCA- (deoxycorticosterone acetate) and TGF-beta-treated renal fibroblasts. Finally, we compared Egr-1 gene expression in DOCA/high salt-induced fibrotic kidneys and untreated mice. Egr-1 transfection of TK188 fibroblasts induced the expression of TIMP-1 and osteopontin mRNA. Similar results were obtained after DOCA-activation of TK188 cells. Stimulation of TK188 with TGF-beta, but not with DOCA, resulted in elevated Coll1A1/Coll1A2 and CTGF levels. Co-stimulation with DOCA and TGF-beta was followed by enhanced Egr-1, Coll1A1, TIMP-1, and CTGF transcription. In conclusion, both DOCA and TGF-beta alone or in combination synergistically induced Egr-1 expression by human renal fibroblasts. DOCA induction of TIMP-1/osteopontin is Egr-1 dependent, whereas TGF-beta appears to induce Coll1A1 and CTGF by an Egr-1 independent pathway. In vivo analyses revealed significantly higher Egr-1 transcript levels in DOCA/high salt-induced fibrotic kidneys compared to untreated mice. Thus, we show for the first time that Egr-1 might participate in DOCA-induced renal fibrosis.