Sahir Kalim

Carbamylation of Serum Albumin as a Risk Factor for Mortality in Patients with Kidney Failure

Increased carbamylation of serum albumin is associated with increased mortality in patients with kidney failure. Counteracting Carbamylation: A Possible Route to Treating Complications of Kidney Failure Like a canary in a coal mine, too much glycated hemoglobin in the blood is a warning sign that a diabetic patients’ glucose is out of control. Carbamyl groups on another ubiquitous blood protein, albumin, may sound a similar alarm for blood urea, a consequence of failing kidneys. Carbamylation may be harmful in its own right because it has been linked to atherosclerosis and other diseases. By using mass spectroscopy, the authors devised a highly accurate assay for measuring carbamylation of the lysine at position 549 of human albumin. With this assay, they found that in two groups of patients with end-stage renal disease and elevated blood urea, the amount of carbamylated albumin correlated with urea concentrations. Albumin was more carbamylated in patients with end-stage renal disease who died within a year than in those who lived longer. Although knowing the prognosis of patients with kidney failure would let us prioritize transplants for the most needy, it would be even better if we could prevent the complications of kidney failure altogether. The authors have gathered data from biochemical experiments and from mice that suggest that protein carbamylation and its associated pathology might be prevented by boosting amino acid concentrations (which tend to be low in these patients) in the blood through diet or other means. Amino acids can compete with protein side chains during carbamylation, potentially interfering with the reaction and its harmful consequences. Thus, amino acid replacement might help patients with kidney failure. Urea, the toxic end product of protein catabolism, is elevated in end-stage renal disease (ESRD), although it is unclear whether or how it contributes to disease. Urea can promote the carbamylation of proteins on multiple lysine side chains, including human albumin, which has a predominant carbamylation site on Lys549. The proportion of serum albumin carbamylated on Lys549 (%C-Alb) correlated with time-averaged blood urea concentrations and was twice as high in ESRD patients than in non-uremic subjects (0.90% versus 0.42%). Baseline %C-Alb was higher in ESRD subjects who died within 1 year than in those who survived longer than 1 year (1.01% versus 0.77%) and was associated with an increased risk of death within 1 year (hazard ratio, 3.76). These findings were validated in an independent cohort of diabetic ESRD subjects (hazard ratio, 3.73). Decreased concentrations of serum amino acids correlated with higher %C-Alb in ESRD patients, and mice with diet-induced amino acid deficiencies exhibited greater susceptibility to albumin carbamylation than did chow-fed mice. In vitro studies showed that amino acids such as cysteine, histidine, arginine, and lysine, as well as other nucleophiles such as taurine, inhibited cyanate-induced C-Alb formation at physiologic pH and temperature. Together, these results suggest that chronically elevated urea promotes carbamylation of proteins in ESRD and that serum amino acid concentrations may modulate this protein modification. In summary, we have identified serum %C-Alb as a risk factor for mortality in patients with ESRD and propose that this risk factor may be modifiable with supplemental amino acid therapy.

A Plasma Long-Chain Acylcarnitine Predicts Cardiovascular Mortality in Incident Dialysis Patients

Background The marked excess in cardiovascular mortality that results from uremia remains poorly understood. Methods and Results In 2 independent, nested case‐control studies, we applied liquid chromatography‐mass spectrometry‐based metabolite profiling to plasma obtained from participants of a large cohort of incident hemodialysis patients. First, 100 individuals who died of a cardiovascular cause within 1 year of initiating hemodialysis (cases) were randomly selected along with 100 individuals who survived for at least 1 year (controls), matched for age, sex, and race. Four highly intercorrelated long‐chain acylcarnitines achieved the significance threshold adjusted for multiple testing (P<0.0003). Oleoylcarnitine, the long‐chain acylcarnitine with the strongest association with cardiovascular mortality in unadjusted analysis, remained associated with 1‐year cardiovascular death after multivariable adjustment (odds ratio per SD 2.3 [95% confidence interval, 1.4 to 3.8]; P=0.001). The association between oleoylcarnitine and 1‐year cardiovascular death was then replicated in an independent sample (n=300, odds ratio per SD 1.4 [95% confidence interval, 1.1 to 1.9]; P=0.008). Addition of oleoylcarnitine to clinical variables improved cardiovascular risk prediction using net reclassification (NRI, 0.38 [95% confidence interval, 0.20 to 0.56]; P<0.0001). In physiologic profiling studies, we demonstrate that the fold change in plasma acylcarnitine levels from the aorta to renal vein and from pre‐ to post hemodialysis samples exclude renal or dialytic clearance of long‐chain acylcarnitines as confounders in our analysis. Conclusions Our data highlight clinically meaningful alterations in acylcarnitine homeostasis at the time of dialysis initiation, which may represent an early marker, effector, or both of uremic cardiovascular risk.

Protein Carbamylation in Kidney Disease: Pathogenesis and Clinical Implications

Carbamylation describes a nonenzymatic posttranslational protein modification mediated by cyanate, a dissociation product of urea. When kidney function declines and urea accumulates, the burden of carbamylation naturally increases. Free amino acids may protect proteins from carbamylation, and protein carbamylation has been shown to increase in uremic patients with amino acid deficiencies. Carbamylation reactions are capable of altering the structure and functional properties of certain proteins and have been implicated directly in the underlying mechanisms of various disease conditions. A broad range of studies has demonstrated how the irreversible binding of urea-derived cyanate to proteins in the human body causes inappropriate cellular responses leading to adverse outcomes such as accelerated atherosclerosis and inflammation. Given carbamylations relationship to urea and the evidence that it contributes to disease pathogenesis, measurements of carbamylated proteins may serve as useful quantitative biomarkers of time-averaged urea concentrations while also offering risk assessment in patients with kidney disease. Moreover, the link between carbamylated proteins and disease pathophysiology creates an enticing therapeutic target for reducing the rate of carbamylation. This article reviews the biochemistry of the carbamylation reaction, its role in specific diseases, and the potential diagnostic and therapeutic implications of these findings based on recent advances.

Does Vitamin D Modulate Blood Pressure

Purpose of review Both vitamin D deficiency and hypertension are highly prevalent. It is unclear whether vitamin D modulates blood pressure and therefore whether vitamin D testing and therapy should become part of routine hypertension prevention and management. This article provides an overview of the data, with special emphasis on the work published in the last 2 years. Recent findings Several animal studies corroborate the strong effect of vitamin D on the renin–angiotensin–aldosterone axis. Small and large observational studies have found associations between vitamin D, increased blood pressure, and the risk of developing hypertension. In contrast, recent data from randomized trials are mixed. Two randomized trials with approximately 1 year of follow-up detected no association between vitamin D treatment and blood pressure, whereas another study of active vitamin D reported a 9-mmHg decrease in systolic blood pressure. Meta-analyses have linked vitamin D levels with blood pressure, but the effect of vitamin D administration on blood pressure remains controversial. Summary Vitamin D deficiency is asociated with high blood pressure in observational studies. This effect is thought to be partly mediated through regulation of the renin–angiotensin–aldosterone axis. However, randomized clinical trials and their meta-analyses have yielded inconclusive results. Large randomized trials focusing on patients with severe vitamin D deficiency and hypertension are needed before vitamin D can be recommended for the prevention or treatment of hypertension.

Nutritional Vitamin D Supplementation in Dialysis: A Randomized Trial

BACKGROUND AND OBJECTIVES Vitamin D (25-hydroxyvitamin D; 25[OH]D) deficiency is common in patients initiating long-term hemodialysis, but the safety and efficacy of nutritional vitamin D supplementation in this population remain uncertain. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS This randomized, placebo-controlled, parallel-group multicenter trial compared two doses of ergocalciferol with placebo between October 2009 and March 2013. Hemodialysis patients (n=105) with 25(OH)D levels ≤32 ng/ml from 32 centers in the Northeast United States were randomly assigned to oral ergocalciferol, 50,000 IU weekly (n=36) or monthly (n=33), or placebo (n=36) for a 12-week treatment period. The primary endpoint was the achievement of vitamin D sufficiency (25[OH]D >32 ng/ml) at the end of the 12-week treatment period. Survival was assessed through 1 year. RESULTS Baseline characteristics were similar across all arms, with overall mean±SD 25(OH)D levels of 21.9±6.9 ng/ml. At 12 weeks, vitamin D sufficiency (25[OH]D >32 ng/ml) was achieved in 91% (weekly), 66% (monthly), and 35% (placebo) (P<0.001). Mean 25(OH)D was significantly higher in both the weekly (49.8±2.3 ng/ml; P<0.001) and monthly (38.3±2.4 ng/ml; P=0.001) arms compared with placebo (27.4±2.3 ng/ml). Calcium, phosphate, parathyroid hormone levels, and active vitamin D treatment did not differ between groups. All-cause and cause-specific hospitalizations and adverse events were similar between groups during the intervention period. Lower all-cause mortality among ergocalciferol-treated participants was not statistically significant (hazard ratio, 0.28; 95% confidence interval, 0.07 to 1.19). CONCLUSIONS Oral ergocalciferol can increase 25(OH)D levels in incident hemodialysis patients without significant alterations in blood calcium, phosphate, or parathyroid hormone during a 12-week period.

Protein carbamylation is associated with heart failure and mortality in diabetic patients with end-stage renal disease

Serum carbamylated albumin (C-Alb) levels are associated with excess mortality in patients with diabetic end stage renal disease. To gain insight into the pathophysiology of carbamylation, we determined associations between C-Alb and causes of death in patients on chronic hemodialysis. The Die Deutsche Diabetes Dialyse Studie (4D study) was a randomized controlled trial testing the effects of atorvastatin on survival in diabetic patients on dialysis during a median follow-up of 4 years. We stratified 1,161 patients by C-Alb to see if differences in carbamylation altered the effects of atorvastatin on survival. Baseline C-Alb significantly correlated with serum cardiac stress markers troponin T and N-terminal pro-B-type-natriuretic peptide, and was associated with history of heart failure and arrhythmia. C-Alb was strongly associated with 1-year adjusted risk of CV mortality, sudden cardiac death and the 4-year risk of death from congestive heart failure (Hazard Ratios of 3.06, 3.78 and 4.64, respectively), but not with myocardial infarction or stroke. Patients with low C-Alb, treated with atorvastatin, experienced a significant improvement in their 4-year survival (Hazard Ratio 0.692). High C-Alb levels are associated with ongoing cardiac damage, risk of congestive heart failure and sudden cardiac death. Thus, carbamylation and uremic cardiomyopathy are associated in patients with diabetes mellitus and kidney disease. Additionally, statins were specifically beneficial to hemodialysis patients with low C-Alb.

Carbamylation of Serum Albumin and Erythropoietin Resistance in End Stage Kidney Disease

BACKGROUND AND OBJECTIVES The mechanisms underlying erythropoietin resistance are not fully understood. Carbamylation is a post-translational protein modification that can alter the function of proteins, such as erythropoietin. The hypothesis of this study is that carbamylation burden is independently associated with erythropoietin resistance. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS In a nonconcurrent prospective cohort study of incident hemodialysis patients in the United States, carbamylated albumin, a surrogate of overall carbamylation burden, in 158 individuals at day 90 of dialysis initiation and erythropoietin resistance index (defined as average weekly erythropoietin dose [U] per kg body weight per hemoglobin [g/dl]) over the subsequent 90 days were measured. Linear regression was used to describe the relationship between carbamylated albumin and erythropoietin resistance index. Logistic regression characterized the relationship between erythropoietin resistance index, 1-year mortality, and carbamylation. RESULTS The median percent carbamylated albumin was 0.77% (interquartile range=0.58%-0.93%). Median erythropoietin resistance index was 18.7 units/kg per gram per deciliter (interquartile range=8.1-35.6 units/kg per gram per deciliter). Multivariable adjusted analysis showed that the highest quartile of carbamylated albumin was associated with a 72% higher erythropoietin resistance index compared with the lowest carbamylation quartile (P=0.01). Increasing erythropoietin resistance index was associated with a higher risk of death (odds ratio per unit increase in log-erythropoietin resistance index, 1.69; 95% confidence interval, 1.06 to 2.70). However, the association between erythropoietin resistance index and mortality was no longer statistically significant when carbamylation was included in the analysis (odds ratio, 1.44; 95% confidence interval, 0.87 to 2.37), with carbamylation showing the dominant association with death (odds ratio for high versus low carbamylation quartile, 4.53; 95% confidence interval, 1.20 to 17.10). CONCLUSION Carbamylation was associated with higher erythropoietin resistance index in incident dialysis patients and a better predictor of mortality than erythropoietin resistance index.

Acute Kidney Injury in HIV-Infected Patients

Acute kidney injury is common in human immunodeficiency virus (HIV)-infected patients, and has been associated with increased morbidity and mortality. Before the introduction of effective antiretroviral therapy, acute kidney injury in HIV-positive patients was most commonly the result of volume depletion, septicemia, or nephrotoxic medications. Acute kidney injury remains a significant problem in the antiretroviral era, and still commonly is attributed to infection or nephrotoxic medications. Less common causes such as direct infectious insults, immune restoration inflammatory syndrome, rhabdomyolysis, and obstruction should be considered when the underlying process is not obvious. In addition to advanced HIV disease, several other patient characteristics have emerged as potential risk factors for acute kidney injury in the antiretroviral era, including older age, diabetes, pre-existing chronic kidney disease, and hepatitis co-infection or liver disease.

An overview of renal metabolomics

The high-throughput, high-resolution phenotyping enabled by metabolomics has been applied increasingly to a variety of questions in nephrology research. This article provides an overview of current metabolomics methodologies and nomenclature, citing specific considerations in sample preparation, metabolite measurement, and data analysis that investigators should understand when examining the literature or designing a study. Furthermore, we review several notable findings that have emerged in the literature that both highlight some of the limitations of current profiling approaches, as well as outline specific strengths unique to metabolomics. More specifically, we review data on the following: (i) tryptophan metabolites and chronic kidney disease onset, illustrating the interpretation of metabolite data in the context of established biochemical pathways; (ii) trimethylamine-N-oxide and cardiovascular disease in chronic kidney disease, illustrating the integration of exogenous and endogenous inputs to the blood metabolome; and (iii) renal mitochondrial function in diabetic kidney disease and acute kidney injury, illustrating the potential for rapid translation of metabolite data for diagnostic or therapeutic aims. Finally, we review future directions, including the need to better characterize interperson and intraperson variation in the metabolome, pool existing data sets to identify the most robust signals, and capitalize on the discovery potential of emerging nontargeted methods.

Early life history transitions and recruitment of Picea mariana in thawed boreal permafrost peatlands

Black spruce (Picea mariana) is the most abundant tree species in the boreal biome, but little is known about how climate warming may change recruitment in peatlands, especially those affected by permafrost thaw. We used results from a seven-year study in northern Manitoba, Canada, to address the following questions: (1) What is the relative importance of early life history transitions on P. mariana recruitment? (2) How are these transitions mediated by biological and environmental factors, including competition, facilitation, disease, herbivory, water table depth, and soil nutrients? (3) Do interactions among these factors create additional recruitment limitations beyond those imposed by environmental factors changing with climate warming, such as hydrology? Seed rain was measured over six years on forested permafrost plateaus and in neighboring collapse scar bogs. Seed germination and seedling survival and growth were measured over 4-5 years in collapse scars and assessed across a three-level water table treatment. Survival and growth experiments examined additional combinations of above- and belowground vascular plant competition and fertilizer addition. Results showed that failure of germination and survival on growing moss surfaces and reduced survival of seedlings in wetter microsites were primary constraints. Seed influx was significantly lower in collapse scars but likely did not limit recruitment. Biological and environmental factors mediating these life history transitions also differed in relative importance, and interactions among them tended to amplify recruitment limitation. Seedling survival was most strongly controlled by fast-growing mosses in wet microsites but also was influenced by apparent drowning in wet plots, herbivory, and loss of foliage caused by a fungal pathogen. Seedling growth was strongly controlled by water table depth, nutrient and competition levels, and fungal pathogens. Multiple, interacting factors will affect P. mariana establishment in boreal peatlands during climate warming. Generalizations about recruitment relying on few environmental gradients sensitive to climate change, such as water table, may therefore not fully capture the complexities of establishment.